Poultry
Both layers and broilers have been studied extensively with Effective Microorganisms®. After all, chicken is the number one meat consumed in the world and chicken eggs are the number one egg consumed in the world. Above are links to the latest research we could find on the internet. TeraGanix is conducting research as well and we will load those papers once we have them completed.
“Effective Microorganisms®” (EM®) in Reducing Noxiousness of Selected Odorant Sources
Environment Protection Engineering
Vol. 36 2010 No. 1
ANDRZEJ KULIG*, RADOSŁAW BARCZAK*
The article presents the changes in a chemical quality of air and in the olfactory noxiousness levels caused by the application of Effective Microorganisms® (EM) contained in the EM-Farming™ preparation. The preparation was applied in selected facilities considered to be substantial sources of odorants. The research was conducted in a wastewater treatment plant, a composting plant, at a waste landfill and in a poultry house.
Within the framework of the project, basic indicators of the olfactory effect, namely ammonia and hydrogen sulphide, were examined along with carbon dioxide, the latter used as the indicator of the intensity of biochemical processes, including the odorigenic ones. Moreover, the sensory assessment method was employed to characterise the olfactory effect organoleptically.
The analysis of the results showed that the negative impact of the facilities on the surrounding area changed within a limited range of values. No clear upward/downward trends shown by the examined gases were observed. For each facility a different and unique pattern of the changes in the concentrations of NH3, H2S and CO2 as well as in odour intensity was detected.
1. INTRODUCTION
A substantial progress in reducing the emissions of selected gaseous or dust pollutants into the atmosphere has been achieved in Poland in the past twenty years, but the emission of odorants continues to be a serious problem in certain industries, municipal service facilities and intensive animal breeding [4]. In the case of a waste water treatment plant, a waste dumping site or a composting plant, an air-tight sealing or full separation of the facility from its surroundings may pose some technical and technological problems and additionally it can be economically unjustified [14]. However, in animal breeding facilities (poultry house, cow shed, pigsty or stable), the use of some methods reducing the impact of odorants, e.g. the masking of odours by chemical means, can be hazardous due to its potentially negative influence on the livestock.
* Environmental Protection and Management Division, Faculty of Environmental Engineering, War- saw University of Technology, ul. Nowowiejska 20, 00-653 Warsaw, Poland, the corresponding author: Andrzej.Kulig@is.pw.edu.pl; tel. +48-22-629-30-26.
According to data received from Central and Provincial Environmental Protection Inspection Offices, Sanitary and Epidemiological Stations and other organizations (e.g. local government bodies) in Poland the highest number of complaints concerning the animal breeding and husbandry relate to the operation of poultry keeping, swine and slaughter/milk cattle breeding facilities. And the complaints about the olfactory noxiousness of municipal service facilities focus predominantly on municipal waste water treatment plants and waste dumping sites [4], [7]. Due to the fact that the number of composting plants have been growing continuously it can be expected that the number of complaints against them will grow, as well.
Because of the difficulties encountered in the process of controlling the olfactory noxiousness, it is necessary to look for effective solutions, as universal as possible, or at least most effective in the given groups of facilities. The research in the field has concentrated on various methods; biological methods are also explored, including those employing the so-called effective microorganisms (EM technology). A potentially positive influence of EM on minimisation of the olfactory noxiousness has been demonstrated, e.g. for leachates from organic waste composting processes [8] and a kitchen waste composting process [9].
2. GOAL AND SCOPE OF THE RESEARCH
The goal of the research was to establish the influence of the consortium of microorganisms (of the EM type) contained in the EM Farming™ preparation on the level of the olfactory noxiousness of selected facilities.
The research was conducted in facilities involved in provision of municipal services, namely:
a) a wastewater treatment plant where a pile of screenings from the plant was examined;
b) a composting plant where piles of compost produced in Dano technology were examined;
c) a solid waste dumping site where a leachate catch pit and a ventilation well were examined as well as in a facility used for animal production purposes, namely a poultry house of about 1000 m2, designed for 20,000 chickens.
The selected facilities can be numbered among those emitting especially disagreeable odours and being most complained about and it is extremely difficult to reduce their odorant emission levels effectively.
Time schedule for the research of each facility investigated was individual: for dumping site – 3 rounds of tests were carried out during 10 weeks, for waste water treatment plant – 4 rounds in one month, and for composting plant as well as for poultry house – 5 rounds during 3–4 months. Air samples were taken in each facility prior to the introduction of the microorganisms to establish a point of reference or a research background to be used for interpretation of the examination results. Two similar compost piles were compared in the composting plant: the first one was treated with the EM preparation, whereas at the second one the composting processes were not modified. For methodological reasons the examination of this facility could show the real influence of the microorganisms on the change in the concentrations of the gases examined.
In the research process, the concentrations of ammonia, hydrogen sulphide and carbon dioxide in air were determined. These gases are the basic indicators of the impact of the facilities under examination on their immediate surroundings [5]. They were chosen in order to obtain information about the processes (generating, among other impacts, disagreeable odours) taking place in the facilities before and after ap- plying of the microorganisms. An organoleptic profile of the olfactory impact has been developed by means of the sensory assessment method. Each time, in the process of taking the air samples, the air temperature and humidity and, in the case of the open-air facilities, the wind velocity and direction were measured.
3. METHODOLOGY
3.1. EM PREPARATION CHARACTERISTICS
The EM preparation is a consortium of a few dozen strains of various microorganisms, both aerobic and anaerobic ones, chosen by Teruo Higa, professor of horticul- ture [2]. The mixture of microorganisms includes, among others, photosynthetic bacte- ria (Rhodopseudomonas palustris, Rhodobacter sphaeroides), Actinomycetes, lactic acid bacteria and fungi (e.g. yeast). The microorganisms produce substances which enable them to survive in unfavourable environmental conditions; consequently, they can effectively compete with destructive and pathogenic microorganisms and to re- place them. The EM preparation contains numerous enzymes which can decompose the organic matter in an environmentally-friendly manner and ensure the survival and growth of the microorganisms both in the soil and in other environmental media [1].
3.2. PREPARATION DOSING METHODOLOGY
The EM preparation was injected into the screenings and the compost piles, poured into the wells collecting leachate or gas from waste and applied in the form of mist (spraying) or by sprinkling within the buildings. In the screenings storage yard, the
EM preparation was for the first time applied on 8.10.2007 in the amount of 5 dm3 per 1 m3 of the screenings; for the second time the preparation was applied on 29.10.2007 in the amount of 2 dm3 per 1 m3. The compost pile was treated with the preparation for the first time on 8.10.07 (3 dm3 per 1 Mg of the compost pile) and for the second one – on 26.11.07 (1.5 dm3 per 1 Mg). 3 dm3 of the preparation were poured both into the leachate catch pit and into the ventilation well on 28.11.07, on 3.12.07 in the amount of 15 dm3, on 8.12.07 in the amount of 10 dm3, and on 24.01.08 in the amount of 3 dm3. In the poultry house, the preparation was applied in the form of mist or by sprin- kling. The sprinkling took place on 24.10, 28.10 and 15.11.07 (0.05 dm3 per 1 m2 of the poultry house floor area), whereas the preparation mist was sprayed on 27.10, 29.10, 3.11, 6.11, 17.11 and 4.12.07 (0.0083 dm3 per 1 m3 of the poultry house cubature).
3.3. MEASURING DEVICES
An aspiration method was employed to determine the concentration of pollutants in the air. The method consists in passing a certain volume of air through a filter made up of the washers connected in series. The washers contained absorb- ing solutions adequately selected for each pollutant. A four-station AG-4 aspirator with the capacity of 60 dm3/h was employed to collect certain amounts of the air containing the gases examined. Within the framework of the methodology adopted, some efforts were taken to minimise the influence of weather conditions, especially the wind, on the sample collection. In the case of the surface source of odorants at the open-air facilities, i.e. the screenings pile and the compost pile, the air samples were collected from under a special 60-dm3 bowl put on the piles (figure 1). The method of sample collection from under the flux chamber used in the facilities directly affected by weather conditions made it possible to minimise the impact of determined substances potentially blown in by the wind and coming from other sources.
Fig. 1. Schematic diagram of the air sampling by flux chamber from a surface-based source
The washers were connected to the AG-4 aspirator and the air from the locations examined was fed to the washers via small hoses. In the catch pits, the hose ends were placed at the depth of 1.2 m. The pits were covered, therefore the influence of external factors was negligible. In the poultry house, the hoses were fixed on a tripod to take in the air on a constant level of 1.3 m above the ground.
The wind velocity was measured by means of A-1200 E anemometer. The air temperature and humidity were measured using the Assman psychrometer.
3.4. METHODOLOGY APPLIED TO DETERMINE POLLUTANT CONCENTRATIONS IN THE AIR
The air samples were collected by passing the air through absorbing solutions. In the case of ammonia, the absorbing solution was 0.01 n sulphuric acid, 2% zinc acetate (Zn(CH3COO)2) was used as the absorbing solution for hydrogen sulphide, and the solution of 0.05 n barium hydroxide (Ba(OH)2) – for carbon dioxide [6]. In most measurements, the time of air sample collection was 30 minutes. In order to adapt the sample size to the absorbing capacity of the absorbing solution, in some cases the collection time was shortened.
The ammonia concentration in the air was determined on the basis of the modified standard PN-Z-04041:1971 [6], [12], and the hydrogen sulphide concentration – on the basis of the modified standard PN-Z-04015-13:1996 [6], [13].
Carbon dioxide, being a gas of an acid character (carbonic acid anhydride), is absorbed by alkaline solutions. The relatively best absorbing agent of carbon dioxide contained in the air is the solution of Ba(OH)2. As a result, barium carbonate is formed. The excess of barium hydroxide taken for absorption was determined by its titration with hydrochloric acid in the presence of phenolphthalein indicator [6].
In the poultry house, the air samples were taken simultaneously at 4 different measuring points. Due to a relatively homogeneous character of the entire poultry house the data used for the analysis were averaged. Similar conditions existed in the screenings storage yard and the compost piles; the only difference was that the samples were taken simultaneously at 2 points at each place.
3.5. ODORIMETRIC MEASUREMENTS
The olfactory influence of odorant sources was characterized organoleptically by means of a sensory assessment of odour intensity i, based on the Just scale expanded by one grade by KULIG (table 1) [3], [5]. The sensory assessment was carried out by a team of two people subject to an odour intensity perceptibility test [10].
Table 1
The organoleptic sensory assessment of the olfactory impact of the compost and screenings piles was carried out at a distance of 1 m from the odorant source. In the poultry house, the assessment was made inside the room, and in the case of the leachate catch pit and the ventilation well the assessment was carried out at their outlets.
4. RESULTS AND DISCUSSION
The research was conducted in the period from October 2007 to February 2008. The results of meteorological measurements carried out in order to assess the odour impact are presented in table 2 and the changes of gaseous pollutant concentrations are shown in figures 2–7.
Table 2
The analysis of the results shows that the intensity of a negative impact of odorants on the surrounding area varies within a limited range of values. During the research no clear upward/downward trends in the concentrations of gases and odour intensity were observed. Each odorant source has its own unique pattern of changes.
A substantial decrease in carbon dioxide concentration was observed in the screenings storage yard: from the initial value of 36.4 g/m3 recorded on 8.10.2007 prior to the application of EM to 7.2 g/m3 the next day following a 24-hour-long expo- sure to the preparation; to 2.4 g/m3 on 15.10.2007; to 1.4 g/m3 recorded on the last day of the research (7.11.2007). Ammonia concentration followed the same pattern: its concentration decreased from 1 350 μg/m3 to 283 μg/m3; to 83 μg/m3 and to zero at the end of the research (figures 2 and 3). The odour intensity decreased almost in the en- tire range covered by the research. The hydrogen sulphide concentrations were negligibly low (below the threshold value).
Fig. 2. Average concentration of CO2 and odour intensity in the air above the surface of the screenings storage yard
Fig. 3. Average concentration of NH3 and odour intensity in the air above the surface of the screenings storage yard
The first three measurements at the waste water treatment plant were carried out under quite similar weather conditions, as in October the air temperature ranged from 7.0 °C to 10.0 °C. Wind direction and velocity did not change much, either: the winds from south-western direction prevailed and their velocity varied from 1.1 to 1.4 m/s. The last measurement, on 7.11.2007, was taken under slightly different weather conditions: the air temperature dropped to 4.6 °C, the wind was stronger (3.2 m/s) but it blew from the south-west again (table 2).
It is not possible to determine unambiguously the influence of the effective microorganisms on the reduction of both the olfactory noxiousness and the concentrations of the gases due to a lack of an arbitrary reference point. However, a positive impact of EM on the quantitative reduction of the odorants emitted by the screenings storage yard cannot be excluded, especially at early stages where the application of the preparation was followed by almost fivefold decrease in the carbon dioxide and ammonia emissions.
The ammonia emissions decreased substantially at the compost pile treated with EM preparation as compared with the reference pile providing the background. Ammonia concentration decreased from an initial 423 μg/m3 for the background and 358 μg/m3 for the pile (prior to the application of EM) to respectively 158 and 8.5 μg/m3 recorded in the last day of the research (5 February 2008). As in the case of the screenings storage yard, ammonia concentration decreased almost threefold following a 24-hour exposure to the EM preparation. A downward trend in ammonia concentration was also recorded in the reference pile, but this value dropped just twofold (figure 4). The increase in the ammonia concentration recorded in both piles on 11 December 2007 (as compared with the previous determinations made on 22.10.2007) was caused probably by a relatively high temperature in winter time (above 6.0 °C). In the re- search period, no hydrogen sulphide emissions were recorded at the piles.
There is another factor which makes this result supports the application of EM: the EM-treated pile is slightly younger (1 month at the maximum), therefore a higher ammonia concentration (compared with the research background) should hold longer.
However, the sensory odorimetric examination did not revealed any differences between the piles. The intensity of odour emitted by both piles decreased gradually from an initial i = 3÷3.5 prior to the application of EM, through i = 2.5÷3 recorded on 9.10.2007, i = 2 recorded on 22.10.2007; i = 0÷2 recorded on 11.12.2007 to i = 0÷1 in the last day of the research (5.02.2008). The differences in odour intensity recorded in particular days depend on the place in the pile where organoleptic sensory assessment is made.
Some positive changes in the concentrations of the gases of interest were also observed in the leachate catch pit and the ventilation well of the waste dumping site. Both in pit and well, the odour intensity decreased although it was still on quite a high level. Also the concentration of hydrogen sulphide decreased from the initial reference value of 23.3 mg/m3 (recorded on 28 November 2007) to 20 mg/m3 on 11 December 2007 and 6 mg/m3 on 5 February 2008 at the leachate catch pit, and from the initial reference value of 1.7 mg/m3 to 0.8 mg/m3 and zero concentration of hydrogen sulfide recorded in the last day of the research at the ventilation well (figures 5 and 6). Moreover, the leachate catch pit showed reduced concentration of carbon dioxide: from 15.5 g/m3 to 13.9 g/m3 and 10.1 g/m3 (figure 5).
Fig. 4. Average concentration of NH3 in the air above the surface of the compost piles
Fig. 5. Concentration of CO2 and H2S in the leachate catch pit at the waste dumping site
The research was conducted in autumn and winter, hence relatively low ambient temperature, especially at the moment of collecting the first samples from pit and well (0.1 °C on 28 November 2007), could affect biochemical processes [11]. However, the leachate catch pit and the ventilation well are built into the waste dumping site a dozen or so metres deep. The conditions (e.g. thermal ones) inside them hardly change with time (especially in a two-month period) and depend on the processes taking place, with a similar intensity, inside and in the lower part of the waste pile. It can be assumed that such a significant change in the concentration was not only a result of natural processes taking place in the waste dumping site, but it also de- pended on the microorganisms introduced that changed both concentration and odour intensity.
Fig. 6. Concentration of H2S in the ventilation well at the waste dumping site
Fig. 7. Average concentration of CO2 and NH3 in the air in the poultry house
In the poultry house, the effect of the preparation was limited mainly to the compen- sation of the effects connected with an increased pollutant emission caused by the gain in the total chicken mass and an increase in their droppings. An increase in the chicken mass and in the amount of droppings results in a growing concentration of the gases in the entire period of the poultry keeping cycle. After an initial systematic increase in the ammonia concentrations from 2.2 mg/m3 recorded on 29.10.2007, 4.9 mg/m3 recorded on 7.11.2007 and 12.2 mg/m3 recorded on 8.11.2007 up to the maximum of 18.2 mg/m3 recorded on 21.11.2007, a decrease in its concentration to 12.2 mg/m3 was observed in the last day of the research (6.12.2007). A similar upward trend followed by a decrease in the concentration was shown by carbon dioxide, but the difference was that its con- centration started to fall as early as on 21.11.2007 from the maximum of 9.8 g/m3 (re- corded on 8.11.2007) to 8.6 mg/m3 (figure 7). As chicken grew the room was aired more frequently and the air temperature decreased from the initial value of 29.5 °C recorded in the first day of the research to about 21 °C recorded in the last day. Hence, the down- ward trend showed by the concentration could have resulted from the action of EM or been caused by an intensified ventilation of the room.
The processes of organic matter degradation taking place in the screenings storage yard or compost piles were extremely intense, especially in the first phase, and could go on even at low temperatures. Nevertheless, the end of our research coincided with the phases of less intense transformations and the period of relatively low temperatures which could have resulted in a diminished activity of the microorganisms and a slow- down in the biochemical processes. It is also important to point out that in the poultry house, in order to ensure an adequate growth of the poultry, the temperature did not fall below 20 °C; therefore it can be assumed that in this case any external factors had no significant influence on the development and activity of the effective microorganisms. Taking account of the above it can be concluded that in the majority of places examined, microbiological processes were effective, although less intensive than in summer time.
5. CONCLUSIONS
The research made it possible to determine the changes in the olfactory noxiousness in the places of interest due to the activity of the consortium of microorganisms (of the EM type) contained in the EM-Farming™ preparation. It can be presumed that consortia of microorganisms are capable of reducing odorant emissions but their ac- tivity is specific, depending on individual place. The methodology employed made it possible to obtain only partially unambiguous, repeatable in the future results. The preliminary research was conducted over the least favourable seasons, i.e. in autumn and winter (in the period from October 2007 to February 2008). In the winter period, the decomposition processes are much less intense than in summer. In spite of an exothermic character of the majority of processes which produce, among other com- pounds, ammonia, hydrogen sulphide and carbon dioxide, low temperatures effectively slow down or even prevent the activity of some organisms, e.g. those which produce hydrogen sulphide. Low temperatures limit substantially the intensity of bio- chemical transformations and, probably, affect EM, as well. The research should be continued with a stricter control of operating conditions, e.g. room ventilation operations, and paying close attention to the technological work which has an effect on the odorant emission levels.
REFERENCES
[1] GRABAS M., CZERWIENIEC E., TOMASZEK J., MASŁOŃ A., LESZCZYŃSKA J., Effectiveness of the proc- essing of sludge with bio-preparation EM-bio and structural material, Environ. Prot. Eng., 2009, Vol. 35, No. 2, 131–140.
[2] HIGA T., Rewolucja w ochronie naszej planety, Fundacja Rozwój SGGW, Warsaw, 2003.
[3] JUST J., Odory i metody ich mierzenia w powietrzu, Roczniki PZH, 1970, No. 21, 601–610.
[4] KOŚMIDER J., MAZUR-CHRZANOWSKA B., WYSZYŃSKI B., Odory, Wydawnictwo Naukowe PWN, Warsaw, 2002.
[5] KULIG A., Metody pomiarowo-obliczeniowe w ocenach oddziaływania na środowisko obiektów go-spodarki komunalnej, Oficyna Wydawnicza Politechniki Warszawskiej, Warsaw, 2004.
[6] KULIG A., OSSOWSKA-CYPRYK K., SKORUPSKI W., WIECZOREK B., Proste metody badania zanie-czyszczeń powietrza atmosferycznego, Materiały PZITS, No. 412/7, Warsaw, 1984.
[7] MALANOWSKA A., KULIG A., Analiza problemu oddziaływania zapachowego na podstawie skarg ludności oraz inwentaryzacji źródeł odorantów w gospodarce ściekowej w Polsce oraz na terenie województwa mazowieckiego, VI konferencja Problemy unieszkodliwiania odpadów, Wydział Inży-nierii Chemicznej Politechniki Warszawskiej, Warszawa, 15 grudnia 2008, 58–63.
[8] SCHMIDT C.E., Technical memorandum describes the bench-scale testing that was conducted in or- der to assess the odor control, Executive Summary of the Independent Environmental Consultant Report, California, USA, August 2005.
[9] SEKERAN V., BALAJI C., BHAGAVATHI PUSPHA T., Evaluation of effective microorganisms (EM) in solid waste management, Electronic Green Journal, Issue 21, April 2005, 66–70, University of Idaho Library.
[10] Odor Sensitivity Test Kit, St. Croix Sensory, Inc. Lake Elmo MN USA, 2005.
[11] OSSOWSKA-CYPRYK K., KULIG A., Oddziaływanie zapachowe procesów mikrobiologicznych prze-biegających w obiektach gospodarki komunalnej, Przegląd Komunalny, Zeszyt Komunalny, 2005, nr 10, 75–82.
[12] PN-Z-04041:1971 Oznaczanie zawartości amoniaku w powietrzu.
[13] PN-Z-04015-13:1996 Ochrona czystości powietrza. Badania zawartości siarki i jej związków. Oznaczanie siarkowodoru na stanowiskach pracy metodą spektrofotometryczną.
[14]Zbiór Reguł ATV-DVWK M 204P: Zmniejszanie emisji substancji zapachowych (odorantów) z oczyszczalni ścieków – stan techniki i jej zastosowanie, materiały pomocnicze, Wydawnictwo Seidel-Przywecki, Warszawa, 2003.
Effect of Effective Microorganisms® (EM4) on Health of Layers
Find the article here: Pakistan Journal of Biological Sciences
Z.H. Naqvi, M. Mushtaq-ul-Hassan, Z. Chaudry, M. Adram and R. Ahmad
Abstract: Commercial laying hens (Babcock) 174-weeks old, were given feed containing 1, 2 and 3% EM4 for a period of 12 weeks EM4 ded not influence live body weight. Egg production was greater in birds given feed containing 1 and 2% EM 4 but lower in birds given feed containing 3% EM4 than the control birds. Blood glucose was significantly lesser in birds given feed containing 3% EM4 than the control birds (p<0.01). serum total proteins, serum albumin, serum globulins, serum total lipids and serum cholesterol were not influenced significantly with the EM 4 treatment. The study suggests that EM4 is a safe product for laying birds it increases egg production when mixed in feed.
Effect of Effective Microorganisms® (EM®) on the Growth Parameters of Fayoumi and Horro Chicken
International Journal of Poultry Science 10 (3): 185-188, 2011 ISSN 1682-8356© Asian Network for Scientific Information, 2011
1. Poultry Research, Agricultural Research Center, P.O. Box 32, Debre Zeit, Ethiopia
2. Faculty of Veterinary Medicine, Addis Ababa University, P.O. Box 34, Debre Zeit, Ethiopia
3. International Livestock Research Institute, Addis Ababa, P.O. Box 5689, Addis Ababa, Ethiopia
Abstract:
The study was conducted to determine growth promoter effects of Effective Microorganisms (EM) on Horro and Fayoumi chickens. This study was conducted at Agricultural Research Center and National Veterinary Institute (Debre Zeit). A total of 450 chickens (225 from each breed) were used in this study. Birds were grouped according to treatment groups: EM-treated (with feed, with water, with feed and water) and non-treated (only SRBC treated and Non-EM, Non-SRBC) controls. EM was given daily from the 3rd week of age for 5 weeks. Daily feed intake and weekly body weight measurements were made. The finding shows that: (1) EM supplementation had no observable effect on mortality, weight gain and FCR in both breeds but Fayoumi had inherently higher body weight than Horro. This study did not consider the cellular arm of the immune response and EM response to infections with specific pathogens was not investigated, collectively demanding further research in these and other issues if EM has to be used as good feed additive.
Key words: Effective microorganism, Horro, Fayoumi, growth parameters, hummoral response
Corresponding Author: D. Tadelle, International Livestock Research Institute, Addis Ababa, P.O. Box 5689, Addis Ababa, Ethiopia 185
INTRODUCTION
Poultry rearing is considered to create rural employment, improve nutritional status of the people, generate family incomes and play a significant role in the social, cultural and religions lives of the society (Tadelle and Ogle, 2001; Halima et al., 2009). Despite the huge population and though poultry meat and eggs could be affordable sources of animal protein, the per capita egg and chicken meat consumption in Ethiopia are very low. Major problems in poultry production systems in this regard are disease, low nutrition, poor management and poor genetic performance (Tadelle and Ogle, 2001; Halima et al., 2009).
Available experimental data show that our indigenous birds have limited genetic capacity for both egg and meat production (Negussie, 1999). However, local chickens have several invaluable characteristics appropriate to traditional low input/low output farming systems, which are not found in any exotic breed. These allow them make the best use of locally available resources, hatch their own eggs, brood their offsprings and tolerate, at least to some extent, the common local poultry diseases (Tadelle, 2003).
Studies in other areas have already shown that the use of probiotics and Effective microorganisms could improve growth parameters: feed intake, weight gain, feed conversion ratio in Broilers (Chantsawang and Watcharangkul, 1999; ZuAnon et al., 1998; Patidar and Prajapati, 1999; Ergun et al., 2000; Kumprechtova et al., 2000; Safalaoh, 2006; Jagdish and Sen, 1993; Alvarez et al., 1994; Silva et al., 2000; Vicente et al., 2007; Higgins et al., 2007). But there are also studies which indicated that use of Probiotics and Effective Microorganisms did not bring positive or significant effect in broilers (Vicente et al., 2007; Samanta and Biswas, 1995; Gohain and Sapcota, 1998; Panda et al., 2000; Ergun et al., 2000; Mohit et al., 2007; Ahmad, 2004) with respect to growth parameters and mortality. This study was therefore conducted to study the effect of Effective Microorganisms on the growth parameters and mortality of Horro and Fayoumi chickens.
MATERIALS AND METHODS
Study animals: 500 fresh and clean eggs of less than 7 days were selected from each of the parental groups (Fayoumi and Local Horro) and incubated. The eggs were hatched at the facility of the center strictly following standard hatchery practices. After the hatching, the chicks were checked for presence of any physical deformity that might later on affect the performance of the birds.
Housing and feeding of experimental chicks: All chicks were randomly assigned to experimental pens and reared in floor pens filled with hay as litter material with a density of 6 birds/meters square. The feed were formulated using appropriate feed formulation computer software (Feed-win). The birds were provided with a starter feed and water ad libitum till 8 weeks of age (end of experimental study). Standard bio-security protocol was employed throughout the experimental period; however, the chicks were not vaccinated for any of theprevalent diseases.
Supplementation of Effective Microorganisms (EM®):
The different EM preparations used in this study were made following the guidelines prepared by EM research organization (EMROSA, 2003). Birds were provided with EM with feed, with water or with feed and water daily starting from the age of 3 weeks till the end of the experiment (week 8). For this, 1% EM-Bokashi in feed was made for EM with feed, 0.1% EM-activated solution in water for EM in water and half of the above concentrations for EM supplemented in both feed and water. Control groups (F-NT-C, F-SRBC, H-NT-C and H- SRBC) did not receive EM.
Growth parameters and mortality: Data on the weight gain, feed intake, feed conversion ratio and mortality of chickens were weekly recorded starting from the 4th week of age till the end of the experiment. Body weight was taken by weighing birds of the same replication together and the average was taken as the weight of the group. Feed intake was determined by subtracting the weight of feed refused from that of feed offered for each replication and the average was taken for the group. Feed conversion ratio was determined as the ratio of the amount of feed consumed per weight gain. Mortality was recorded as it occurred.
Statistical analysis and model: The parameters including body weight gain, feed conversion ratio were monitored. All data were analyzed by ANOVA with the repeated model mixed procedure of SAS software (2000) and compared by least square means. Significance of observed mortality was tasted by chi-square statistics. Least square means were considered statistically different at p<0.05.
RESULTS
Growth parameters
Mortality: Mortality of Fayoumi and Horro chickens was recorded throughout the experimental period. There was no significant difference between breeds, between EM- treated and Non-treated controls and between the different modes of EM supplementation for each breed (Table 1).
Weight gain and FCR: The initial weight of chicks on week 4 (immediately before SRBC treatment) varied between individuals in different groups and the two breeds. Therefore, weight gain is presented by subtracting week 4 weight from measurements registered each week after SRBC treatment. Accordingly, in all treated and control groups weight gains show a general linear pattern (Fig. 1). The Fayoumi breed demonstrated significantly higher weight gain than the Horro for both treated and control groups (p<0.05). At week 8 (end of the experiment), supplementation of EM with water caused significantly lower weight gain than controls and other EM modes of supplementation in both breeds at p<0.05. On the other hand, the Feed Conversion Ratio (FCR) showed no regular pattern and no significant variation between groups and between breeds was seen (Data not shown).
Fig. 1: Weekly weight gains (in grams) taking weight at week 4 as reference. Fayoumi (F) and Horro (H) chicken groups supplemented with EM in feed (EM-F), in water (EM-W), in feed and water (EM-FW), non-treated control (C) and SRBC groups are compared
Table 1: Experimental groups of Fayoumi and Horro chickens with different modes of EM treatment. Each group is composed of three replications of 15 birds
Table 2: Total observed mortality of Fayoumi and Horro chicken in different groups during the experimental period (week 4-8)
| Group | No. Chicks/group | No. Birds died |
|---|---|---|
| F-EM-F | 45 | 2 |
| F-EM-W | 45 | 2 |
| F-EM-FW | 45 | 1 |
| F-SRBC | 45 | 6 |
| F-NT-C | 45 | 5 |
| H-EM-F | 45 | 4 |
| H-EM-W | 45 | 3 |
| H-EM-FE | 45 | 4 |
| H-SRBC | 45 | 6 |
| H-NT-C | 45 | 4 |
DISCUSSION
Effect of EM on growth parameters: The mortality rate was not improved after EM supplementation in both Fayoumi and Horro and breed difference was not significant. In agreement with our finding, Yoruk et al. (2004) found that mortality of hens fed with control diet was not different from those fed probiotic diets. During our study period, there was no outbreak of any known disease in which the effect of EM could have been tasted to arrive at a definitive conclusion. Therefore, it is only possible to say that it did not significantly reduce the effects of unidentified causes of mortality during the study period. On the contrary, other studies have shown that probiotics significantly reduced mortality in chickens (Vicente et al., 2007), Salmonella colonization in one day-old broiler chicks (Higgins et al., 2007) and mortality in turkeys (Vicente et al., 2007). The duration of the study may be one of many factors that may have caused differences between the results of ours and others. Recently, in a study in turkey poults with idiopathic diarrhea conducted by Higgins et al. (2007), administration of three doses of this probiotic culture was reported to improve body weight gain similar to the response obtained with therapeutic. Throughout the study period, Fayoumi chicks had higher gain in terms of weight than the Horro chicks. This was true both between treated groups and between control groups. Therefore, it seems that the difference between the two breeds is governed by inherent factors rather than due to the application of EM. The FCR was measured as the ratio of the amount of feed consumed per weight gain. There was no observable variation or appreciable pattern for FCR between breeds, between treatment groups within a breed and between EM treated and non-treated groups. Therefore, it appears that EM has little effect on FCR as both breeds have similar FCR at least during the early periods of their development. Feed conversion ratio as affected by probiotics is the subject of controversy. Similar to the present findings, several previous works suggested that supplementation of probiotics does not influence feed conversion ratio significantly or no such effect on FCR (Samanta and Biswas, 1995; Gohain and Sapcota, 1998; Panda et al., 2000; Ergun et al., 2000; Mohit et al., 2007; Ahmad, 2004). Similarly supplementation of probiotics had no effect on the performance of broiler chicks (ZuAnon et al., 1998; Patidar and Prajapati, 1999; Ergun et al., 2000; Kumprechtova et al., 2000). On the other hand, there are reports which state that EM fed birds had significantly higher weight gain and lower FCR than Non-EM groups (Safalaoh, 2006; Jagdish and Sen, 1993; Alvarez et al.,1994; Silva et al., 2000). EM supplementation had little effect on growth parameters such as weight gain, mortality and FCR. The difference in weight gain between Fyoumi and Horro appear to be due to breed difference rather than due to an EM effect. However, this study was conducted with limited number of chicks and with relatively young birds (less than two months of age). Therefore, to arrive at better understanding and conclusion on the role of EM and its future utilization in poultry farming the following points need to be addressed in future researches. As a novel area of research in our country, the findings of the present study are encouraging. However, it needs to be strengthened by similar works that involve large number of animals for prolonged period of time so that more plausible conclusion could be drawn for its future application in animal farms.
REFERENCES
Ahmad, I., 2004. Effect of probiotic (protexin) on the growth of broilers with special reference to the small intestinal crypt cells proliferation. M. Phil Thesis, Centre of biotechnology, University of Peshawar.
Alvarez, L.C., E.M. Barrera and E.A. Gonzalez, 1994. Evaluation of growth promoters for broiler chickens. Veterinaria Mexico, 25: 141-144.
Chantsawang, S. and P. Watcharangkul, 1999. Influence of effective microorganisms on the quality of poultry products. In: Proceedings of International Conference on Kyusei Nature Farming, October 23-26, 1997, Sara Buri, Thailand, pp: 133-150.
Effective Microorganisms Research Organization of South Africa, 2003. WWW.EMROSA.SA.
Ergun, A., S. Yalcin and P. Sacakli, 2000. The usage of probiotic and zinc bacitracin in broiler rations. Ankara Universities Veteriner Facultesi Dergisi, 47:271-280.
Gohain, A.K. and D. Sapcota, 1998. Effect of probiotic feeding on the performance of broilers. Ind. J. Poult. Sci., 33: 101-105.
Halima, H., F.W.C. Neser, A. De-Kock and E. Van Marle-Köster, 2009. Study on the genetic diversity of native chickens in northwest Ethiopia using microsatellite markers. Afr. J. Biotechnol., 8: 1347-1353.
Higgins, J.P., S.E. Higgins, V. Salvador, A.D. Wolfenden, G. Tellez and B.M. Hargis, 2007. Temporal effects of lactic acid bacteria probiotic culture on Salmonellain neonatal broilers. Poult. Sci., 86: (in press).
Jagdish, P. and A.K. Sen, 1993. Effect of different growth promoters on the performance of broilers. Poult. Advisory, 26: 49-51.
Kumprechtova, D., P. Zobac and I. Kumprecht, 2000. The effect of Sacchromyces cervisiae SC47 on chicken broiler performance and nitrogen output. Czech J. Anim. Sci., 45: 169-177.
Mohit, A.H., S.A. Hosseini, H. Lotfollahian and F. Shariatmadari, 2007. Effects of probiotics, yeast, vitamin E and vitamin C supplements on performance and immune response of laying hen during high environmental temperature. Int. J. Poult. Sci., 6: 895-900.
Negussie, D., 1999. Evaluation of the performance of local, Rod Island Red and Fayuomi breeds chicken under different management regimes in the highlands of Ethiopia. M.Sc. Thesis, Swedish University of Agriculture Sciences, Uppsala.
Panda, A.K., M.R. Reddy, S.V.R. Rao, M.V.L.N. Raju and N.K. Praharaj, 2000. Growth, carcass characteristics, immunocompetence and response to Escherichia coli of broilers fed diets with various levels of probiotic. Archive fur Geflugelikunde, 64: 152-156.
Patidar, S.K. and J.B. Prajapati, 1999. Effect of feeding Lactobacilli on serum antibody titer and faecal microflora in chicks. Microbiologic Aliments Nutr.,17: 145-154.
Safalaoh, A.C.L., 2006. Body weight gain, dressing percentage, abdominal fat and serum cholesterol of broilers supplemented with a microbial preparation. Afr. On Line J. Food, Agric. Nutr. Dev., 6: 1-10.
Samanta, M. and P. Biswas, 1995. Effect of feeding probiotic and lactic acid on the performance of broiler. In. J. Poult. Sci., 30: 145-147.
SAS software, 2000. version. 8. SAS Institute Incorporated. North Carolina.
Silva, E.N., A.S. Teixeira, A.G. Bertechini, C.L. Ferreira and B.G. Ventura, 2000. Ciencia e Agrotecnologia. 24: Ed. Especial, 224-232.
Tadelle, D., 2003. Phenotypic and genetic characterization of local chicken ecotypes in Ethiopia, Ph.D. Dissertation, Der Humboldt-University Berlin, pp: 2-208.
Tadelle, D. and B. Ogle, 2001. Nutritional status of village poultry in the central highlands of Ethiopia. Trop. Anim. Health Prod., 33: 521-537.
Vicente, J.L., L. Aviña, A. Torres-Rodriguez, B. Hargis and G. Tellez, 2007. Effect of a Lactobacillus sp.-based probiotic culture product on broiler chicks performance under commercial conditions. Int. J. Poult. Sci., 6: 154-156.
Yoruk, M.A., M. Gul, A. Hayirli and M. Macit, 2004. The effects of supplementation of humate and probiotic on egg production and quality parameters during the late laying period in hens. Poult. Sci., 83: 84-88.
ZuAnon, J.A., S. Fonseca, H.S. Rostagno, M. Almeida and M. Silva, 1998. Effects of growth promoters on broiler chicken performance. Revista Brasileira de Zootecnia, 27: 999-1005.
Effect of Effective Microorganisms® on Growth Parameters and Serum Cholesterol Levels in Broilers
Wondmeneh Esatu1, Adey Melesse1 and Tadelle Dessie2*
African Journal of Agricultural Research Vol. 6(16), pp. 3841-3846, 18 August, 2011 Available online at http://www.academicjournals.org/AJAR
DOI: 10.5897/AJAR11.176
ISSN 1991-637X ©2011 Academic Journals
Full Length Research Paper
1. Livestock Research Directorate, National Poultry Research Case Team, Debre Zeit Agricultural research Center, Ethiopian Institute of Agricultural Research, Ethiopia.
2. International Livestock Research Institute, Addis Ababa, Ethiopia.
Accepted 12 April, 2011
This study was conducted to evaluate the effect of different administration methods of effective microorganisms (EM®) on the performance and serum cholesterol level of broilers at Debre Zeit Agricultural Research Center, Ethiopia. Uniform weight of mixed sex day-old-broilers of cobb-500 strain (n = 240) were randomly distributed to 4 treatment groups with 3 replications of 20. They were kept under a standard management condition for 49 days being subjected to treatment rations since Day 10 on. Performance parameters were recorded and analyzed. Total blood cholesterol was analyzed with standard kit at the end. The result showed that there was no significant difference of EM administration methods (p < 0.05) on mortality of chickens during the starter (1 to 29 days) and finisher (30 to 49 days) phases. Feed consumption was found to be significantly higher for Treatment 4 (Bokashi in feed + EM in water) than the rest of the treatment groups. Weight gain was significantly higher (p < 0.05) for Treatment 4 (Bokashi +EM in water), during the entire period than the rest of the treatment groups. Birds fed with T4 (Bokashi +EM in water) required less feed for a unit increase in weight during the starter and finisher phase. Birds fed with T3 (Non Bokashi +EM in water) required the highest feed for a unit increase in weight. EM application in all forms has resulted in significantly lower (p < 0.05) total blood cholesterol, EM application both in feed and water combined being most effective in lowering the total blood cholesterol than the other application methods.
Key words: Bokashi, broiler, performance, application, feed conversion ratio, mortality.
*Corresponding author. E-mail: T.DESSIE@cgiar.org.
INTRODUCTION
EM (Effective microorganisms) is the mixed-cell culture which is composed of photosynthetic bacteria, actinomycetes, yeast, lactobacillus and fungi (Higa, 1993). Effective microorganisms (EM) as a new technological advance were innovated in Japan. Effective microorganism (EM) is a combination of 70 to 80 different types of “good” and beneficial microorganisms contributing to the wide range of applications. Microorganisms in EM are not genetically engineered, but they are commonly found in everyday food and healthy forest soils. The microorganisms mutually co-exist to form beneficial relationships with each other in a liquid medium. Microorganisms in EM assist one another for survival in a food chain system and thereby form a synergy that fights off pathogens and rotting microorganisms. EM is self sterilizing (pH between 3.4 to 3.7); therefore, pathogens cannot survive in EM (EMROSA, 2006). The principal organisms of are usually five. They are photosynthetic bacteria (phototrophic bacteria), lactic acid bacteria, yeasts, actinomycetes and fermenting fungi. Photosynthetic bacteria are independent self supporting microorganisms. These bacteria synthesize useful substances from secretions of roots, organic matter and/or harmful gases (example, hydrogen sulfide) by using sunlight as sources of energy. The useful substances comprise of amino acids, nucleic acids, bioactive substances and sugars, all of which promote plant growth and development. These metabolites act as substrates for bacterial growth. Thus, increasing photosynthetic bacteria, which enhance other effective microorganisms (Higa, 1993). Lactic acid bacteria produce lactic acid from sugars, and other carbohydrates produced by Photosynthetic bacteria and yeast. Thus, food and drinks, such as yogurt and pickles have been made by using Lactic acid bacteria for a long period of time. However, lactic acid is a strong sterilizer. It suppresses harmful microorganisms and increases rapid decomposition of organic matter. Moreover; Lactic acid bacteria enhance the breakdown of organic matter such as lignin and cellulose, and ferment these materials without causing harmful influences arising from un- decomposed organic matter (Higa and Parr, 1994). The yeasts that are present in effective microorganisms have a wide range of functions. They produce antimicrobial substances to kill off all harmful pathogens. In addition, they also produce beneficial substances, such as hormones, enzymes and vitamin B. Their secretions are useful substrates for effective microorganisms such as lactic acid bacteria and actinomycetes. Yeasts synthesize antimicrobial and useful substances from amino acids and sugars secreted by photosynthetic bacteria, organic matter and etc. Actinomycetes, the structure of which is intermediate to that of bacteria and fungi, produces antimicrobial substances from amino acids secreted by photosynthetic bacteria and organic matter. These antimicrobial substances suppress harmful fungi and bacteria. Actinomycetes can co-exist with photosynthetic bacteria. Thus, both species enhance the quality of the soil environment, by increasing the antimicrobial activity of the soil (Higa, 1993). Fermenting fungi such as Aspergillus and Penicillium decompose organic matter rapidly to produce alcohol, esters and antimicrobial substances. These suppress odors and prevent infestation of harmful insects and maggots. Each component of effective microorganisms (photosynthetic bacteria, lactic acid bacteria, yeasts, actinomycetes and fermenting fungi) has its own important function. However, photosynthetic bacteria are the pivot of EM activity. Photosynthetic bacteria support the activities of other microorganisms. On the other hand, photosynthetic bacteria also utilize substances produced by other microorganisms. This phenomenon is termed "coexistence and co-prosperity". EM products that are of importance for poultry production are Stock EM, Multiplied EM and Bokashi (solid form of EM). Stock EM is the basic, concentrated EM solution that contains all the beneficial microorganisms. Stock EM is not a fertilizer, a chemical or a synthetic. It is also not genetically engineered. Stock EM is a basic dominant form of EM and is therefore usually used to produce multiplied EM (M-EM). Multiplied EM is the activated, secondary form of EM. It consists of S-EM (1 to 3%), molasses (3 to 5%) and water (94%). The molasses serves as a nutrient source for the microorganism, consequently leading to the growth and multiplication of the microorganisms. Therefore the name, multiplied EM. EM Bokashi is an essential supplement feed for animals and is made from 1 to 2% Multiplied EM, 1% molasses and 98% water, which is then added to organic feed materials. It has various applications, but is mostly used as a form of animal feed (APNAN, 1995).
According to Chantsawang and Watcharangkul (1999), EM was first introduced and used extensively in Asia; the technology was later introduced to other various countries. The use of EM in animal husbandry nowadays is very well identified in many parts of the world. In a study conducted in Belarus by Konoplya and Higa (2000), EM was successfully used in poultry and swine units as feed constituent and sanitation spray. In South Africa EM was used to increase productivity in integrated animal units and poultry farms (Hanekon et al., 2001). Result of EM experiment conducted on 27000 Kuroki broilers in Japan is reported by Higa (1994). According to the author; EM was administered in drinking water, EM Bokashi in feed and sluicing or cleaning out the hen coops with an EM dilute solution. Of the 27000 chicks reared by this operation, survival rate for shipment was only 83% prior to the introduction of EM method; whereas, the survival rate increased to 97% for the birds raised by EM methods. In actual figure, this made an increase of 3780 fowls ready and shipped to market. Tortuero (1973) found that a lactobacillus probiotic and zinc bacitracin had similar effects in stimulating weight gain and feed efficiency of broiler chicks. An alternative is the use of probiotics, prebiotics and symbiotic (feeding probiotic microorganisms together with prebiotic substances) which might contribute due to the development of beneficial microorganisms in the gastrointestinal tract (Pelicano et al., 2004). In human health there is significant evidence that probiotics such as specific types of lactobacillus bacteria and bifido bacteria can lower the three major risk factors for coronary heart diseases and stroke: excessive cholesterol, high blood pressure and high triglyceride levels. Study in Argentina (Taranto, 1999) indicated that Lactobacillus bacteria lowered total blood cholesterol by 22% and triglycerides by 33%. However, research done by (Pelicano et al., 2004; Greany et al., 2004) has shown no benefit of using microbial preparation or probiotics. Therefore, since not all strains of bacteria such as Lactobacillus acidophilus and Lactobacillus bulgaricus work to lower cholesterol level, note worthy care should be taken with strain selection of beneficial bacteria to get the best results. The other benefit of EM is that it eliminates odors without producing odorous gases by dominating the microbial ecology with organisms that exploit a fermentative pathway (Yongzhen and Weijiong, 1994).
The present study was therefore conducted to add information on this relatively new technology (introduced by Japanese Professor, Dr. Teuro Higa in 1980); the effect of EM on growth parameters and total serum cholesterol level of broilers.
MATERIALS AND METHODS
Description of the study area
The study was conducted in Debre Zeit Agricultural Research
Table 1. Composition and nutrient content of starter and finisher.
| Nutrient | Starter (0-29 days) | Finisher (30-49 days) |
|---|---|---|
| Metabolizable energy (kcal/kg) | 3200 | 3100 |
| Crude protein (%) | 21.82 | 22 |
| Calcium (%) | 0.59 | 0.56 |
| Phosphorus (%) | 0.43 | 0.41 |
| L-Lysine (%) | 1.13 | 0.84 |
| DL-Methionine (%) | 0.39 | 0.32 |
| Methionine + cystein (%) | 0.71 | 0.70 |
Center (DZARC), poultry research farm, which is located at 47 km south east of Addis Ababa. The elevation of the area is around 1920 m above sea level. The average temperature of the area is 16.6°C with the mean minimum and maximum annual temperature of 8.9 and 24.3°C respectively. The area has two rainy seasons; the minor one is between February and April and the major one is from June to September. The annual mean rainfall is about 851 mm (DZARC, 2003).
Experimental design and treatments
The experiment was carried out from September 4 to October 23, 2009. Mixed sex day-old-broiler chickens of cobb-500, bought from local hatchery. Uniform weight of mixed sex day-old-broilers of cobb-500 strain (n = 240) were randomly assigned to 4 treatment groups with 3 replications of 20. EM was made and mixed with compounded standard feed to make EM in feed and with water to make EM treatment in a solution form. The Bokashi (EM in a solid form) and the EM activated solution (EM a solution form) were prepared following the procedures of Asia Pacific natural agriculture Network (APNAN, 1995). EM in a feed was prepared by adding 3% of Bokashi (EM in a solid form) mixed to the feeds to make up Bokashi added feed and 2 ml of activated EM solution was added in a liter of drinking water. Using the Bokashi and activated EM solution, 4 feeds types (treatments) were prepared. Treatment 1 control: a diet with no EM in feed + EM in water, Treatment 2: EM in drinking water only, Treatment 3: EM in feed only and Treatment 4: EM in feed and EM in drinking water.
Management of experimental animals
Standard commercial management was followed during the trial. The chicks were housed in an open house, concrete floor, in 12 pens with 20 chicks per pen with a density of 10 birds per square meter. Saw dust sprayed with standard disinfectant was used as litter material with about 7 mm thickness. Round feeders and drinkers of small sizes were used during the starter phase (0 to 29 days) throughout the trial. Infrared lamp of 250 W was used for the first week. Bigger size drinker of 10 L and round hanging type feeder of 12 kg were used during the finisher phase. (30 to 49 days). Internal temperature and humidity was controlled by opening and closing the curtains to keep the chickens comfortable. The chickens were vaccinated with Newcastle (HB1 at Day 7 and Lasota at Day 21) and Gumboro at Day 10. Treatment feeds and water were given ad-libitum to the chickens throughout the experiment period starting Day 10 onwards.
Experimental diets
Experiential diets were formulated to contain, metabolizable energy
(ME) 3200 kcal/kg and 21% crude protein (CP) during the starter phase (1 to 29 days); and ME 3100 kcal/kg and 22% CP for finishers (30 to 49 days) using trial and error feed formulation software. The nutritive content as per the analysis of the sample is presented in (Table 1).
Parameters studied
Data were recorded during the periods from 1 to 29 days and 30 to 49 days. Feed intake was determined by subtracting the quaintly leftover from the quantity offered every day. Weight gain was measured every day and gain was calculated every week as a difference between final and initial weight. Feed conversion ratio was determined by calculating the ratio of feed consumed per a unit of growth. Mortality was calculated as the ratio of death occurrences per the remaining animals multiplied by 100. Serum cholesterol was also analyzed.
Sample collection
At the end of the experiment 3 ml of blood sample from each slaughtered broiler chickens (which were fed EM with feed, EM with water, EM with feed and water, and control groups) was collected from the jugular vein by using plain vacutainer tube for total cholesterol analysis. The collected blood was allowed to clot over night (12 h) in a vertical position to separate the serum. The collected sera were stored at -20°C until analysis. Total cholesterol assay was done using cholesterol liquicolour commercial kit (Human Diagnostics Worldwide) based on CHOD-PAP method according to the manufacturer’s instruction at faculty of veterinary medicine, Debre Zeit laboratory.
Statistical analysis
Statistical software (SAS, 2000) was used to analyze the data. Differences between treatment means were evaluated by Duncan’s multiple range tests at a significance level of 5%.
RESULTS
Feed intake and weight gain
Feed intake was significantly higher (p< 0.05) for broilers fed EM in feed and water followed by those fed with control diet during the starter phase (Table 2). During the finisher phase, the same parameter was found to be significantly higher (p< 0.05) for the birds supplemented
Table 2. Production performance of broilers as affected by EM supplementation in drinking water and feed: Starting phase (0 to 29 days) (n = 200, x ± SD).
| EM Application | ||||
|---|---|---|---|---|
| Variable | Control | EM in drinking water | EM in feed | EM in feed and drinking water |
| Weight gain (g) | 744 ab | 724ab | 706ab | 780a |
| Feed intake (g) | 1391.57b | 1347.83C | 1334.93c | 1513.2a |
| FCR(amount of feed required/a unit gain in weight in Kg) | 1.87a | 1.86a | 1.89a | 1.94a |
| Mortality (%) | 5.0a | 6.0a | 4.0a | 5.0 |
Means in the same row with different superscripts were significantly different at P< 0.05.
Table 3. Production performance of broilers as affected by EM supplementation in drinking water and Feed: Finisher phase (30 to 49 days) (n = 200, x ± SD).
| EM Application | ||||
|---|---|---|---|---|
| Variable | Control | EM in drinking water | EM in feed
| EM in feed and drinking water |
| Weight gain (g) | 1789bc | 1681bc | 1724bc | 2143a |
| Feed intake (g) | 4151.73b | 4068.3c | 4016.57c | 4306.87a |
| FCR(amount of feed required/a unit gain in weight in Kg) | 2.32b | 2.42c | 2.33b | 2.01a |
| Mortality (%) | 2.0a | 2.0a | 4.0a | 2.0a |
Means in the same row with different superscripts were significantly different at P< 0.05.
with EM in feed and water, followed by those fed control diet (Table 3). Weight gain of the broilers fed with EM in feed and water was significantly higher (p< 0.05), followed by those fed with EM in feed alone during the starter phase (Table 2). But during finisher phase, weight gain of the birds fed with EM in feed and water was found to be significantly higher (p< 0.05) than the rest of the treatment groups including the control diet.
during the starter phase. And during the finisher phase, the same was found to be true for the birds supplemented with EM in feed and water. However, in similar study by Chatsavang and Watchangkul (1999) showed no significant differences in the feed intake among the treatment groups during the starter and finisher phases. In this study weight gain of the broilers fed with EM in feed and water was significantly higher, followed by those fed with EM in feed alone during the starter phase. But during finisher phase, significant weight gain was observed on those birds which were fed with EM in feed and water. However, in similar work done by Chatsavang and Watchangkul (1999), EM supplementation in feed only resulted in significantly higher body weight. (p< 0.05), but during the finisher phase all treatments did not reveal any significant difference was observed with respect to mortality during the finisher phase (T able 3).
Total serum cholesterol level
Total cholesterol level of blood taken from broilers fed with control diet (no EM supplemented) showed significantly higher than groups fed with EM in different forms at (p< 0.05) (Table 4).
Feed conversion ratio and mortality
DISCUSSION
There was no significant difference among the treatment groups and mortality during the starter phase (Table 2). Birds fed with EM in feed and water required lower feed for a unit increase in weight (p< 0.05), while no significant difference
Feed intake and weight gain
Feed intake was higher for boilers fed EM in feed and water followed by those fed with control diet among themselves. In addition, study by ZuAnon et al. (1998), Patidar and Prajapati (1999), Ergun et al. (2000) and Kumprechtova et al. (2000) stated that supplementation of probiotics had no effect on the performance of broiler chicks. On the other hand, there are reports which state that EM fed birds had significantly higher weight gain and lower FCR than Non-EM groups (Safalaoh, 2006; Jagdish and Sen, 1993; Alvarez et al.,1994; Hamid et al., 1994).
Table 4. Total serum cholesterol level (n = 80, x ± SD).
| EM Application | ||||
|---|---|---|---|---|
| Variable | Control | EM in drinking water | EM in feed | EM in feed and drinking water |
| Serum cholesterol(mg/dl) | 131.33 ± 3.17a | 102.94 ± 8.7b | 103.81 ± 13.9b | 103.51 ± 14.7b |
Means in the same row with different superscripts were significantly different.
Feed conversion ratio and mortality
Total cholesterol level of blood taken from broilers fed with control diet (no EM supplemented) showed significantly higher than groups fed with EM in different forms. This is of course in line with a result reported somewhere else by Taranto (1999) that EM application has resulted in lowering excessive cholesterol, high blood pressure and high triglyceride levels in humans. The ability of probiotics to depress serum cholesterol content has been reported in broilers and rats (Mohan et al., 1996; Grunewald, 1982). Similarly, though not significantly different (P < 0.05), the level of serum cholesterol at 42 days was lower for the EM supplemented birds than for the control (Safalaoh, 2006). However, the results of current and previous studies are different from that of other studies that showed that probiotic supplementation had no beneficial effects on broiler performance and did not lower cholesterol levels in post-menopausal women (Pelicano et al.,2004; Greany et al., 2004).
Total serum cholesterol level
In terms of mortality, no significant difference was seen in the starter phase and finisher phase among the treatment groups. This is same in line with the finding by Y oruk et al. (2004) that mortality of hens fed with control diet was not different from those fed with probiotic diets. On the contrary, other studies have shown that probiotics significantly reduced mortality in chickens (Vicente et al., 2007). Several previous works suggested that supplementation of probiotics does not influence feed conversion ratio significantly or no such effect on FCR (Samanta and Biswas, 1995; Gohain and Sapcota, 1998; Panda et al., 2000; Ergun et al., 2000; Mohit et al., 2007; Ahmad, 2004). In addition, study by Chatsav ang and W atchangkul (1999) did not reveal any significant difference among the treatment groups.
ACKNOWLEDGMENTS
The authors wish to express their gratitude to the Ethiopian Institute of Agricultural Research (EIAR) for facilitating the experiment and DZARC poultry research staff for their unreserved effort up to the completion of the research. We also are thankful for EM Woljjejji plc for provision of EM stalk solution.
CONCLUSION AND RECOMMENDATION
According to the result of the current study, EM application in various forms can positively affect weight gain, feed intake and lowers FCR. It has been shown that EM application can also lowercholesterol level in Broilers. The results of these three experiments show that EM has positive effect on broilers in terms of body weight gain. This study has demonstrated that supplemen- tation of broiler diets with a microbial preparation, such as EM, may offer potential benefits to the poultry industry, such as improvements in body weight gain and feed utilization efficiency. Due to variations among previous studies conducted elsewhere, further research is required to clearly indicate the effect of EM supplementations in poultry diets.
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Mohan B, Kardirvel R, Natarajan A, Bhaskaran J (1996). Effect of probiotic supplementation on growth, nitrogen utilization and serum cholesterol in broilers. Br. Poult. Sci., 37(2): 395-402.
Mohit AH, Hosseini SA, Lotfollahian H, Shariatmadari F (2007). Effects of probiotics, yeast, vitamin E and vitamin C supplements on performance and immune response of laying hen during high environmental temperature. Inter. J. Poult. Sci., 6(12): 895-900.
Panda AK, Reddy MR, Rao SVR, Raju MVLN, Praharaj NK (2000). Growth, carcass characteristics, immunocompetence and response to Escherichia coli of broilers fed diets with various levels of probiotic. Arch. fur Geflugelikunde, 64:152-156.
Patidar SK, Prajapati JB (1999). Effect of feeding Lactobacilli on serum antibody titer and faecal microflora in chicks. Microbiol. Aliments Nutr., 17: 145-154.
Pelicano ERL, Souza PA, Souza HBA (2004). Performance of broilers fed diets containing natural growth promoters. Revista Brasileira de Ciência Avícola, 6(4): 231-236.
Safalaoh ACL (2006). Body weight gain, dressing percentage, abdominal fat and serum cholesterol of broilers supplemented with a microbial preparation. Afr. J. Food Agric. Nutr. Dev., 6(1): 1-10.
Samanta M, Biswas P (1995). Effect of feeding probiotic and lactic acid on the performance of broiler. Indian J. Poult. Sci., 30: 145-147.
SAS software (2000). Version.8. SAS Institute Incorporated. North Carolina.
Taranto M (1999). Effect of Lactobacillus reuteri on the prevention of hypercholesterolemia in mice. J. Dairy Sci., 83: 401-403.
Toruero F (1973). Influence of Lactobacillus Acidophilus in chicks on the growth, feed conversion, malabsorption of fats syndrome and intestinal flora. Poult. Sci., 52: 197-203.
Vicente JL, Aviña L, T orres-Rodriguez A, Hargis B, T ellez G (2007). Effect of a Lactobacillus Spp-Based Probiotic Culture Product on Broiler Chicks Performance under Commercial Conditions. Int. J. Poult. Sci., 6(3): 154-156.
Yongzhen N, Waijiong L (1994). Report on the Deodorizing Effect of Effective Microorganisms (EM) in Poultry Production. Beijing, China, 73: 402-407.
Yoruk MA, Gul M, Hayirli A, Macit M (2004). The effects of supplementation of humate and probiotic on egg production and quality parameters during the late laying period in hens. Poult. Sci., 83: 84-88.
ZuAnon JA, Fonseca S, Rostagno HS, Almeida M, Silva M (1998). Effects of growth promoters on broiler chicken performance. Revista Brasileira de Zootecnia, 27: 999-1005.
Effect of Feeding Diets Containing an Antibiotic, or a Probiotic on Grown and Pathogenic Intestinal Bacteria in Domestic Fowls
M. H. El-Deep, K. Amber* and M. A. M. Sayed
Animal Prod. Inst. Agric. Res..Center , Ministry of Agric, Egypt.. Poultry Production Dept, Faculty of Agric, Kafr El-Sheikh Univ, Egypt*.
A 4-wks study was conducted to determine the effect of feeding diets containing an antibiotic (Zinc bactracin) , or a probiotic (Effective Microorganisms ), on performance, and urease Pathogenic Intestinal Bacteria of Inshas chicks (a local Egyptian chicken strain) .The experimental design consisted of six experimental groups: control and 5 dietary treatments as follows; (T1) Basal diet ( control ), (T2) Basal diet + EM (2.5 ml/kg diet ), (T3) Basal diet + EM (5.0 ml/kg diet ), (T4) Basal diet + EM (7.5 ml/kg diet ), (T5) Basal diet + EM (10.0 ml/kg diet ) and (T6) Basal diet + Zinc bactracin (500 mg/kg). Feeding treatment was started at 4 wks of age and lasted at 41 wks of age. Characteristic investigations were including : Live body weight ; Body weight gain ; feed consumption ; efficiency of feed utilization and Bacteria Enumeration (Aerobic plate count (x106/g), E. coli, Salmonella, Staphylococci and Coccidia ovum). Feeding diets containing the probiotic were significantly (P < 0.01) increasing the average daily gain during the expermental period compared to the control . This increase was partially accounted with increased feed intake. During the expermental period, feeding the diet containing probiotic significantly reduce the counts of total viable bacteria (P < 0.01),, E. coli, salmonella, staphylococci and Coccidia ovum in caecum compared with untreated control diets.. Our study indicating that, dietary probiotic decreases pathogenic intestinal bacteria of chicks and this may be beneficial for improving animal health and growth performance.
One way is to use specific feed additives or dietary raw materials to favorably affect animal performance and welfare, particularly through the modulation of the gut microbiota which plays a critical role in maintaining host health [1]. A balanced gut microbiota constitutes an efficient barrier against pathogen colonization, produces metabolic substrates (e.g. vitamins and short-chain fatty acids) and stimulates the immune system in a non- inflammatory manner.
In this context probiotics, prebiotics and synbiotics could be possible solutions. The main effects of these feed additives are the improved resistance to pathogenic bacteria colonization and enhanced host mucosa immunity; thus resulting in a reduced pathogen load, an improved health status of the animals [2] and a reduced risk of food-borne pathogens in foods.
The impact of biotechnology in poultry nutrition is of significant importance. Biotechnology plays a vital role in the poultry feed industry. Nutritionists are continually putting their efforts into producing better and more economical feed. Good feed alone will not serve the purpose but its better utilization is also essential. Dietary changes as well as lack of a healthy diet can influence the balance of the microflora in the gut thus predisposing to digestion upsets. A well-balanced ration sufficient in energy and nutrients is also of great importance in maintaining a healthy gut. A great deal of attention has recently been received from nutritionists and veterinary experts for proper utilization of nutrients and the use of probiotics for growth promotion of poultry.
Effective Micro-organisms (EM) is a microbial preparation developed by Professor T. Higa of University Of The Ryukyus in Japan. The EM is composed of different microbes that include bacteria, yeasts and/or fungi. Some of the benefits claimed to accrue from the use of EM include improved meat and manure quality, improved animal health, reduction of foul smells and absence of toxic effects on bird growth [3]. Use of EM in Africa is a new innovation and novel idea. There is no available literature regarding use of microbial preparations in broiler production. Therefore, this experiments was designed to investigate the possibility of using probiotic namely, (EM) effective microorganism ( instead of using antibiotics) to Inshas chickens (Egyptian local strain ) , and to evaluate its effects on growth and pathogenic intestinal bacteria .
A total number of 540 unsexed vaccinated Inshas (local Egyptian chicken strain) one day-old-chicks were weighed , wing banded and randomly divided into six experimental groups ( three replicates each group ) . The birds were placed in a room (floor pens) maintained at a constant temperature of 28+/-3 oC and a relative humidity of 70+/-3% .Food and water were always available ad libtum . The basal diet was formulated to meet the nutrient
needs suggested by the NRC, 1994 [13]. Body weight was determined individually to the nearest gram at four weeks intervals up to the 40 week of age. Feed intake was recorded every week by supplying a weighed amount of feed and subtracting the unconsumed portion from the total amount offered. The average feed intake of each bird was calculated by dividing the monthly consumed feed by the
INTRODUCTION
MATERIAL AND METHODS
number of individuals in each group during this month , considering the death if any. At the time of slaughter test, 6 samples of ileum and caecum contents were collected and examined to define and count the pathogenic bacteria for each treatment. Fecal matter samples were collected in sterile polyethylene bags. All samples were delivered directly to the laboratory for bacterial count and definition using the procedure of [4]. The experimental design consisted of six dietary treatments as follows; (T1) Basal diet ( control ), (T2) Basal diet + EM (2.5 ml/kg diet ), (T3) Basal diet + EM (5.0 ml/kg diet ), (T4) Basal diet + EM (7.5 ml/kg diet ), (T5) Basal diet + EM (10.0 ml/kg diet ) and (T6) Basal diet + Zinc bactracin (500 mg/kg).. The results obtained were statistically analyzed using Duncan’s Multiple Range Test {17}. Statements of statistical significance are based on P<0.05.
RESULTS AND DISCUSSION
Growth Performance
Data presented in Fig ( 1 ) show the effect of the different dietary treatments on live body weight (LBW) at various ages.
Generally , it could be seen that the feed additives were significantly ( P ≤0.001) increased LBW in all treatments. The results indicated that body weight in all supplemented groups was increased ( P ≤0.001) by 14.4, 15, 16.9, 17.9 and 9.4% for T2, T3, T4, T5 and T6 diets, respectively, as compared with control group. Also, feed intake and feed conversion were improved by feed additives .These results may be explained the review that probiotics are natural control method that based on ensuring the bird has an adequate gut microflora counter pathogenic bacteria colonization in its digestive tract and consequently has healthy gut that results in good digestion and nutrient absorption [5].
Bacteria Enumeration
Results presented in Fig ( 2, 3, 4, 5 and 6 ) indicated that experimental diets caused severe suppression pathogenic intestinal bacteria counts. Where, there were significantly (P<0.01) reduction in counts of total viable bacteria, E. coli, salmonella, staphylococci and Coccidia ovum in caecum comparing with untreated control diet.The present results of EM dietary treatments agrees with that found by [6] who observed that frequency of Salmonella colonization was significantly reduced due to probiotic bacteria treatment. Also, [7] reported that lactobacillus was able to inhibit the growth of some pathogenic bacteria such as E. coli and salmonella. The antagonistic activity of lactic acid bacteria towards pathogens can be attributed to the production of bactericidal substances like bacteriocins, organic acids and hydrogen peroxide as reported by many workers for example [8]. The addition of probiotics product decreased the E. coli count as found by [9]. This type of bacteria produces lactic acid which alters the pH of chicken gut making it improper media for harmful bacteria such as salmonella and pathogenic species of E. coli. [10]. Probiotics decreased proliferation of pathogenic bacteria [11] concluded that probiotics enable the host animal to return to normal through increasing normal gut flora on the expense of pathogenic organisms. Furthermore, [12]reported that the beneficial effect of probiotics since their microbial constituents produce natural lactic acid that helps in maintaining an optimum low pH which inhibits growth of undesirable bacteria leading to optimum enzyme activity. Authors concluded that the antibacterial action produced by probiotics was probably due to a combination of factors which include organic acids (acetic and lactic acids), hydrogen peroxide and bacitracin.
REFERENCES
1. TUOHY, K.M.; ROUZAUD,G.C.M.; BRUCK,W.M.; GIBSON, G.R. (2005): Modulation of the human gut microflora towards improved health using prebiotics-assessment of efficacy. Current Pharmaceutical Design 11: 75–90.
2. CHOCT, M. (2009): Managing gut health through nutrition. British Poultry Science. 50 :9–15.
3. ZHANG, X.; LOU-HUA, LI-JIANLIANG.; XIE-DONGXIA, XU-JIN.; HUO-CUIMEI, CUI-YANSHUN. (2007): A study on the benefit of feeding Hy-line white egg with fermented feed. Southwest-China-Journal-of-Agricultural-Sciences. 20(3): 529-533.
4. A.O.A.C. (1995): Association of official analysis chemists. Official Method Analysis. 16th Ed. Published by A.O.A.C. Benjamin Franklin Station, Washington, D.C.
5. AWAD, W. A.; BOHM, J.; RAZZAZI-FAZELI, E.; FAUKAL, K.; ZENTEK, J.(2003): Effects of feeding deoxynivalenol contaminated wheat on the performance of broiler chickens. Pages 149–153 in Proc. 8. Symp. Eur. Soc.Vet. Comparat. Nutr., Budapest, Hungary.
6. LINE, E.J.; BAILEY, S.J.; COX,N.A.; STERN,N.J.; TOMPKINS, T. (1998): Effect of yeast-supplemented feed on Salmonella and Campylobacter populations in broilers. Poult. Sci. 77: 405-410.
7. OYARZABAL, O.A.; CONNER, D. E. (1995): In vitro fructooligosaccharide untilization and inhibition of salmonella Spp by selected bacteria.. Poult. Sci. 74 : 1418 – 1425.
8. JOERGER M. C.; KLAENHAMMER, T. R. (1986): Characterization and purification of helvetin J and evidence for a chromosomally determined bacteriocin produced by Lactobacillus helveticus 481. J. Bacteriology. 167: 439–446.
9. CHATEAU, N.; CASTELLANOS, I.; DESCHAMPS, A.M. (1993): Distribution of pathogen inhibition in the Lactobacillus isolates of commercial probiotic consortium. J. Appl. Bacteriol. 74: 36-40.
10. LEESSON, S.; MAJOR, D. (1990). As biotechnology grains momentum Canadian researchers study need for feed criteria. Feed stuffs. 62: 23–30.
11. JIN, L. Z.; HO, Y. W.; ABDULLAH, N.; JALALUDIN, S. (1998): Growth performance, intestinal microbial populations and serum cholesterol of broilers diets containing Lactobacillus cultures. Poult. Sci. 77:1259–1265.
12. HUANG, M. K.; CHOI, Y. J.; HOUDE, R.; LEE, J. W.; LEE, B.; ZHAO, X. (2004): Effects of Lactobacilli and an Acidophilic Fungus on the Production Performance and Immune Responses in Broiler Chickens. Poultry Science. 83:788–795.
13. NRC. (1994): Nutrient requirements of poultry. 9th ed. Natinal Acadeny press, Washigton, D.C., U. S. A.
Effect of Probiotic on Gut Development of Domestic Fowls
Kh. Amber, M. H. El-Deep, and M. A. M. Sayed
Animal Prod. Inst. Agric. Res..Center , Ministry of Agric, Egypt. Poultry Production Dept, Faculty of Agric, Kafr El-Sheikh Univ, Egypt*.
This experiment was conducted to study the effects of effective microorganisms (EM) and Zinc bacitracin on gut development, digestibility coefficient of nutrients and intestinal histology of Inshas chicken (a local Egyptian chicken strain) . Five hundredand fourty chicks were randomly assigned to 1 to 6 dietary treatments for 41 wk. The dietary treatments were 1) control; 2) Basal diet + EM (2.5 ml/kg diet ); T3) Basal diet + EM (5.0 ml/kg diet ); T4) Basal diet + EM (7.5 ml/kg diet ); T5) Basal diet + EM (10.0 ml/kg diet ) and T6) Basal diet + Zinc bactracin (500 mg/kg). The obtained results showed that, villi height , villi thickness and villi surface area were significantly increased in birds fed EM with different levels and Zinc bactracin diets . The data on the digestibility coefficient of nutrients revealed that, all nutrients of EM diets were more efficiently digested than that of Zinc bactracin diet (p≤0.01) . While, digestibility coefficient of OM, DM, CP, EE, CF and NFE was significantly increased as compared with chicks kept on the control diet. Moreover, It was generally noticed that intestinal histology was almost following the same trend observed with gut development and digestibility coefficient .
The impact of biotechnology in poultry nutrition is of significant importance. Biotechnology plays a vital role in the poultry feed industry. Nutritionists are continually putting their efforts into producing better and more economical feed. Good feed alone will not serve the purpose but its better utilization is also essential. Dietary changes as well as lack of a healthy diet can influence the balance of the microflora in the gut thus predisposing to digestion upsets. A well-balanced ration sufficient in energy and nutrients is also of great importance in maintaining a healthy gut. A great deal of attention has recently been received from nutritionists and veterinary experts for proper utilization of nutrients and the use of probiotics for growth promotion of poultry.
In poultry nutrition, probiotic species belonging to Lactobacillus, Streptococcus, Bacillus, Bifidobacterium, Enterococcus, Aspergillus, Candida, and Saccharomyces have a beneficial effect on poultry performance [1], modulation of intestinal microflora and pathogen inhibition [2], intestinal histological changes [3], immunomodulation [4], certain haematobiochemical parameters [5], improving sensory characteristics of dressed broiler meat [6] and promoting microbiological meat quality of broilers[7].
Effective Microorganisms (EM) is a microbial preparation developed by Professor T. Higa of University Of The Ryukyus in Japan. The EM is composed of different microbes that include bacteria, yeasts and/or fungi. Some of the benefits claimed to accrue from the use of EM include improved meat and manure quality, improved animal health, reduction of foul smells and absence of toxic effects on bird growth [8]. Use of EM in Africa is a new innovation and novel idea. There is no available literature regarding use of microbial preparations in broiler production.
Therefore, this experiments was designed to investigate the possibility of using probiotic namely, (EM) effective microorganism ( instead of using antibiotics) to Inshas chickens (Egyptian local strain ) , and to evaluate its effects on gut development .
A total number of 540 unsexed vaccinated Inshas (local Egyptian chicken strain) one day-old-chicks were weighed , wing banded and randomly divided into six experimental groups ( three replicates each group ) . The birds were placed in a room (floor pens) maintained at a constant temperature of 28+/-3 oC and a relative humidity of 70+/-3% .Food and water were always available ad libtum. The basal diet was formulated to meet the nutrient needs suggested by the NRC, 1994. For enteric morphometric analysis, birds on the designated evaluation day were euthanized, and small intestines were collected. A 1-cm segment of the midpoint of the lower ileum were removed and fixed in 10% buffered formalin for 72 h. Each segment was then embedded in paraffin, and a 2μm section of each sample was placed on a glass slide and stained with hematoxylin and eosin for examination with a light microscope [9]. The parameters evaluated were villus height, villus and thickness, villus surface area. Morphological parameters were measured using the Image Pro Plus v 4.5 software package. Fourteen measurements were taken per bird per parameter. Villus height was measured from the top of the villus to the top of the lamina propria. Villus surface area was calculated using the formula (2π)(VW/2)(VL), where VW = villus width, and VL = villus length [10], and we used anther way villus surface area index = perimeter length × mucosal thickness [9].The experimental design consisted of six dietary treatments as follows; (T1) Basal diet ( control ), (T2) Basal diet + EM (2.5 ml/kg diet ), (T3) Basal diet + EM (5.0 ml/kg diet ), (T4) Basal diet + EM
INTRODUCTION
MATERIAL AND METHODS
(7.5 ml/kg diet ), (T5) Basal diet + EM (10.0 ml/kg diet ) Multiple Range Test [17]. Statements of statistical and (T6) Basal diet + Zinc bactracin (500 mg/kg).. The significance are based on P<0.05. results obtained were statistically analyzed using Duncan’s
RESULTS AND DISCUSSION
Morphometric Analysis of the Gut
Villi Height
Results obtained showed that villi height was significantly ( p≤0.05) increased in birds fed EM with different levels and Zinc bactracin diets in Fig (1 ). The villi length was longer (P < 0.05) by about 12.6 to 15.2% for birds fed EM and Zinc bacitracin diet as compared with the control. There are no significant differences among chicks fed diets with different levels of EM.
Villi Thickness
Statistical analysis of the results obtained proved that EM and Zinc bacitracin diets had a significant effect on the Villi thickness Fig (1 ). Villi thickness was significantly increased in birds fed EM and Zinc bacitracin diets as compared with control diet ( as average 3.8 vs 1.6 μm; P < 0.001).
Villi Surface Area
Villi surface area as influenced by dietary EM with different levels and Zinc bactracin during the experiment of period is presented in Fig (1 ). Villi surface area was increased (P < 0.001) from 17.11 to 71.3 μm as the level of EM increased from 0 to 10 ml/kg in bird diets. Also, increased (P < 0.001) by 101.3 % in chicks fed T6 diet as compared with those fed control diet . These results agree with [11]. Upon consumption, probiotics deliver many lactic acid bacteria into the gastrointestinal tract. These microorganisms have been reputed to modify the intestinal milieu and to deliver enzymes and other beneficial substances into the intestines [12].
It is well established that probiotics alter gastrointestinal pH and flora to favor an increased activity of intestinal enzymes and digestibility of nutrients [13]. Mechanisms by which probiotics improve feed conversion
efficiency include alteration in intestinal flora, enhancement of growth of nonpathogenic facultative anaerobic and gram positive bacteria forming lactic acid and hydrogen peroxide, suppression of growth of intestinal pathogens, and enhancement of digestion and utilization of nutrients [14].
Illuminating work from Gordon’s laboratory provides evidence that manipulating the microbiota with probiotics can influence the host. Germ-free mice were colonized with Bifidobacteria thetaiotaomicron, a prominent component of the adult human gut microbiota, and Bifidobacterium longum,a commonly used probiotic. B. longum repressed B thetaiotaomicron expression of antibacterial proteins that may promote its own survival in the gut, as well as influence the composition, structure, and function of its microbial community.
Digestibility Coefficients
Data on digestibility coefficient of nutrients as shown in Table ( 1) revealed that all nutrients of EM diets were digested more (p≤0.01) efficiently than of Zinc bactracin diet. While, digestibility coefficient of OM, DM, CP, EE, CF and NFE were significantly increased by 3.7, 3.8, 3.9, 9.6, 4.1 and 4.4 %, respectively, in chicks fed T6 diet. Also, increased (p≤0.01) by 7.3, 7.5, 5.4, 9.0, 53.9 and 7.0 %, respectively, in those fed EM diets ( T2, T3,T4 and T5) as compared with chicks kept on control diet.
[15] reported that supplementing broiler diets with probiotics tended to increase the digestibility of CP in both fresh maize and 10% moldy maize diets. Also, [16] showed that, supplementing broiler chick diets with growth promoter significantly improved digestion coefficient of nutrients except CF compared to unsupplemented diet. The increased number of beneficial microbes was confirmed and explained by [17] who found that the number of anaerobic bacteria and cellulytic bacteria was increased, when the diet was supplemented with yeast, due to enhancing Lactate utilization and moderating pH of the media, therefore, yeast improved the nutrients digestibility coefficients.
In this connection, [18] reported a positive effect of probiotics on apparent protein digestion and attributed this effect to the proteolytic activity of bacteria. It is worthy to note the absence of significant differences in the data as a result of the combination effect of dietary CP level and tested probiotics. Such observation confirmed the previously mentioned opinion that the tested probiotics had a sparing effect of nearly 2.0 % CP. Similarly, the better (P>0.05) digestibility obtained with probiotics supplementation suggests that such addition improved feed and nutrients utilization, which in turn explain the better growth and FCR values obtained with the probiotics supplemented diets. In general, the improvement (P>0.05) due to adding the probiotics may be attributed to improving intestinal microbial balance. In other words, probiotics help to keep the intestinal tract healthy and when the epithelial tissue is healthy, there is improved and better absorption of all nutrients [19].
Improvement of nutrient digestibility by supplementing chick diets with either microbial probiotics could be attributed to different stimulators such as change in the enteric flora and reduction of E. coli population, lowering gastric pH, synthesis of catabolic enzymes that help in releasing cell compounds including amino acids, sugar and fatty, acids into the intestinal environment and involving of active bacteria with the digestive processes, protein synthesis and nutrient absorption in gastrointestinal tract [20]
Fig (1): Morphometric Analysis of the Gut as affected by different levels of EM and Zinc bacitracin.
Table (1): Digestibility coefficient of chicks as affected by the different nutrients of the experimental treatment (Means ± SE).
REFERENCES
1. AWAD, W.A.; GHAREEB, K.; ABDEL-RAHEEM, S.; BOHM, J. (2009): Effects of dietary inclusion of probiotic and synbiotic on growth performance, organ weights, and intestinal histomorphology of broiler chickens. Poult. Sci. 88: 49-56.
2. HIGGINS, J.P.;HIGGINS, S.E.; VICENTE, J.L.; WOLFENDEN, A.D.; TELLEZ, G.; HARGIS, B.M. (2007): Temporal effects of lactic acid bacteria probiotic culture on Salmonella in neonatal broilers. Poultry Science. 86:1662–1666.
3. CHICHLOWSKI, M.; CROOM,W.J.; EDENS, F.W.; MCBRIDE, B.W.; QIU, R.; CHIANG,C.C.; DANIEL, L.R.; HAVENSTEIN,G.B.; KOCI, M.D. (2007): Microarchitecture and spatial relationship between bacteria and ileal, cecal, and colonic epithelium in chicks fed a direct-fed microbial, primalac, and salinomycin. Poult. Sci. 86: 1121-1132.
4. APATA, D.F. (2008): Growth performance, nutrient digestibility and immune response of broiler chicks fed diets supplemented with a culture of Lactobacillus bulgaricus. J. Sci. Food Agric. 88: 1253-1258.
5. ASHAYERIZADEH, A.; DABIRI, N.; ASHAYERIZADEH, O.; MIRZADEH, K. H.; ROSHANFEKR, H., MAMOOEE, M. (2009): Effect of dietary antibiotic, probiotic and prebiotic as growth promoters, on growth performance, carcass characteristics and hematological indices of broiler chickens. Pakis. J. Biol. Sci. 12: 52-57.
6. PELICANO, E.R.L.; SOUZA, P.A.; DESOUZA, H.B.A.; OBA, A.; NORKUS, E.A.; KODAWARA, L.M.; DE LIMA, T.M.A. (2003): Effect of different probiotics on broiler carcass and meat quality. Br. J. Poult. Sci. 5: 207-214.
7. KABIR, S.M.L.; RAHMAN, M.M.; RAHMAN, M.B. (2005): Potentiation of probiotics in promoting microbiological meat quality of broilers. J. Bangladesh Soc. Agric. Sci. Technol. 2: 93-96.
8. ZHANG, X.; LOU-HUA, LI-JIANLIANG.; XIE-DONGXIA, XU-JIN.; HUO-CUIMEI, CUI-YANSHUN. (2007): A study on the benefit of feeding Hy-line white egg with fermented feed. Southwest-China-Journal-of-Agricultural-Sciences. 20(3): 529-533.
9. SAKAMOTO, K.; HIROSE, H.; ONIZUKA, A.; HAGASHI, M.; FUTAMURA, N.; KAWAMURA, Y.; EZAKI, T. (2000): Quantitative study of changes in intestinal morphology and mucus gel on total parenteral nutrition in rats. J. surg. Res. 94: 99-106.
10. SOLIS DE LOS SANTOS, F.; TELLEZ, G;. FARNELL, M. B;. BALOG, J. M.; ANTHONG, N. B. (2005): Hypobaric hypoxia in ascites resistant and susceptible broiler genetic lines influences gut morphology. Poultry science. 84: 1495-1498.
11. MILES, R. D.; BUTCHER, G. D.; HENRY, P. R.; LITTELL, R. C. (2006): Effect of Antibiotic Growth Promoters on Broiler Performance, Intestinal Growth Parameters, and Quantitative Morphology. Poultry Science 85:476–485.
12. MARTEAU, P.; RAMBAUD, J.C. (1993): Potential of using lactic acid bacteria for therapy and immunomodulation in man. FEMS Microbiol. Rev. 12: 207-220.
13. DIERCK, N.A. (1989): Biotechnology aids to improve feed and feed digestion: Enzymes and fermentation. Arch. Anim.Nutr. Berl. 39: 241-261.
14. YEO, J.; KIM, K. (1997): Effect of feeding diets containing an antibiotic, a probiotic, or yucca extract on growth and intestinal urease activity in
broiler chicks. Poult. Sci. 76: 381-385.
15. KIM, C. J.; NAMKUNG, H.; AN, M. H.; PILK, I. K. (1988): Supplementation of probiotics to the broiler diets containing moldy corn. Korean J. of Animal Sci. 30(9); 542-548.
16. ABD EL-SAMEE, M. O.; EL-HUSSEINY, O. M.; ALI, A. M. (2001): Effects of dietary crude protein and Enramy on supplementation of broiler performance. Egypt. Poult. Sci. 21(II): 507-520.
17. DAWSON, K. A. (1987): Mode of action of yeast culture, Yea-Sacc., in the rumen: A natural fermentation modifier. In Biotechnology in the feed industry. T.P. Lysons(Ed)PP. 119-126. Altech Technical Publication, Nicholosville, Kentucky, USA.
18. DE-SCHRIJVER, R.; OLLEVIER , F. (2000): Protein digestion in Juvenile trout and effects of dietary administration of ribrio proteolyticus. Aquaculture. 186:107-116.
19. KAISTHA, M.; KATOCH, S.; KATOCH, B. S.; KUMARI,M.; DOGRA, K. K.; SHARMA, C. R. (1996): Effect of dietary supplementation of useful microbes isolated from luffa cylindrical (luffa aegytiaca) and momordica charantia on the performance of broilers. Indian J. Poult. Sci. 31: 56- 162.
20. ABD EL-AZEEM, F. (2002): Digeston, Neomycin and yeast supplementation in broiler diets under Egyptian summer conditions. Egypt. Poult. Sci. 22: 23– 257.
Effective Microorganisms® (EM®) as an Alternative to Antibiotics in Broiler Diets: Effect on Broiler Growth Performance, Feed Utilisation and Serum Cholesterol
A.C.L. Safalaoh and G. A. Smith
Department of Animal and Wildlife Sciences
University of Pretoria, Pretoria, 002, South Africa
Abstract:
An experiment was conducted to evaluate the effect of using Effective Micro-organisms (EM) as an alternative to antibiotics (AB) on growth performance, feed utilisation and serum cholesterol of broilers. Dietary treatments consisted of supplementation with neither AB nor EM, AB only, EM only or AB plus EM. The EM was supplemented at either 15 g/kg or 30 g/kg while the AB (Zinc Bacitracin) was added at 500 mg/kg. At six weeks of age, birds fed diets with neither the EM nor AB had significantly (P<.05) lower weight gains (2066 g) than the rest of the treatments. Birds fed the diet containing AB and EM at 30 g/kg had significantly (P<.05) higher body weight gain (2096g) than the rest of the treatments. The improvements in BWG were associated with slight enhancement of feed efficiency while the EM effects were more pronounced at the higher dosage (30 g/kg). The poorest feed:gain ratio (1.82) was observed in birds fed diets containing neither EM nor AB. Apart from improving dressing percentage, EM supplementation also resulted in birds with low serum cholesterol levels. This study has shown that EM has growth promoting and hypocholesteremic effects and offers a potential alternative to antibiotics in broiler diets.
Introduction:
Antibiotics (AB) continue to be used in the poultry industry as growth stimulants and therapeutic agents. However, due to the fact that continued use of AB tends to stimulate development of resistance from harmful micro-organisms, there is currently an outcry from the consumer society and health sector to ban their (AB) use as feed additives in animal and poultry feeds. It is therefore urgent and imperative that an alternative to replace antibiotics should be found. However, such an alternative should elicit positive results similar to those of AB without compromising bird growth, feed utilisation and the quality of final product. According to this criteria, and based on the current available knowledge on feed additives, probiotics seem to be the best alternative.
Probiotics have been shown to have growth promoting (Cavazzoni et al, 1998; Jin et al, 1998; Yeo and Kim, 1997; Mohan et al, 1996) prophylactic (Cavazzoni et al, 1998; Yeo and Kim, 1997) and hypocholesteremic effects (Jin et al, 1998; Mohan et al, 1996). Jin et al (1998) attributed enhanced growth rates by probiotics to improved feed efficiency.
There are several probiotics on the market world wide. In South Africa, Effective Micro-organisms (EM)1 is a probiotic that has recently been introduced . Use of EM has been shown to improve animal health (Philips and Phillips, 1996). However, there is paucity of information regarding the efficacy and beneficial effects of EM vis a vis AB. Without any tangible evidence from empirically derived data, the adoption of this innovation in the poultry industry cannot be guaranteed.
This experiment was conducted to evaluate the effects of supplementing broiler diets with EM at two different inclusion rates as an alternative to AB (Zinc bacitracin) on body weight gain (BWG), feed efficiency (FCR), dressing percentage/ carcass yield (CARC) and serum cholesterol (CHOL) of broilers from 1-42 days of age.
Materials and Methods:
Birds
Four hundred and fifty chicks were randomly selected and assigned to 6 treatments. The treatments (Trt.) involved addition of AB at 500 mg/kg (AB) and EM at either 15 g/kg (EM15) or 30 g/kg (EM30). Some diets had no EM (EM0) or AB (AB0) added. The 6 treatments were as follows: Trt.1= EM0, AB; Trt. 2= EM0, AB0; Trt.3=EM15, AB;Trt.4=EM15,AB0;Trt.5=EM30,AB;Trt.6=EM30, AB0.
Each treatment had five replicates of 15 birds each. Birds were housed in an environmentally controlled broiler house with a floor covered with wood shavings for the whole experimental period.
Feed
Birds were fed a commercial starter mash (12.8 MJ/kg ME, 22.99 CP) from 1-28d followed by finisher mash (13.4 MJ/kg ME, 20.03 CP) from 29-42 d of age.
Liquid EM was initially mixed with a portion of the basal diet which contained maize meal, soybean meal, molasses and fish meal to make “Bokashi”. The Bokashi was made as per the method of Phillips and Phillips (1996) before being mixed with the rest of the feed.
1EM Centre, EMROSA (Pty) Ltd, Centurion, Wierdepart, South Africa
Measurements
Bird weight gains and feed intake (FI) measurements were determined at weekly intervals. On day 42, five birds were randomly selected from each treatment for collection of blood through the brachial vein. The blood was drained into a polythene tube and centrifuged at 5,000 rpm for 10 minutes. Serum CHOL was then determined using the Syncron CX System (Beckman Instruments, Inc., 1995). The birds were then killed by cervical dislocation for determination of carcass. The carcass yield excluded feathers, feet, head and viscera and was expressed as percent of live weight and reported as dressing percentage.
Data Analysis
Data were analyzed using the General Linear Models procedures of SAS® (SAS Institute, 1988) at P<.05.
Results
The effect of EM and AB supplementation on BWG, FI and CARC are presented in Table 1.
Birds fed diets containing AB and EM0 (Trt.1) had significantly (P<.05) higher BWG than those fed diets containing neither AB nor EM (Trt. 2) or the diet containing EM15 (Trt. 4). Diets supplemented with AB, EM15 (Trt. 3) produced BWG similar to diets that contained AB, EM30 and AB0, EM30. A combination of AB and EM30 produced the highest BWG. The high BWG was associated with increased FI. Though not significantly (P >. 05) different, feed conversion ratio tended to improve with addition of AB, EM or both. Dressing percentage was significantly (P<.05) higher for birds containing EM30 with or without AB supplementation (Trt.5 and 6).
Effects on serum CHOL are presented in the Fig. 1. Supplementing diets with EM resulted in birds with reduced serum CHOL. The CHOL reducing properties of EM occurred in a dose dependent manner . The lowest serum cholesterol content was observed in birds fed diets containing EM 30.
Two other notable results were observed in this study. A negligible mortality rate of only 0.22 per cent (1 out of 450 birds) occurred for the whole experimental period. The bird died in week 5 (Trt. 5) probably due to Sudden Death Syndrome. Another observation was that during the first few days ( 1-21d), there was a propensity for birds fed diets containing EM diet to have pasted vents.
Measurements
Fig.1. Effect of probiotic and antibiotic supplementation on serum cholesterol of broilers at 42 d of age
Discussion and Conclusions
Diets containing AB, EM0 had better BWG than those with AB0, EM0. This is a clear manifestation and demonstration of the growth stimulation effect of AB.
Addition of EM at 15g/kg elicited no beneficial effects on BWG unless supplemented together with AB. However, an inclusion rate of 30g/kg of EM with or without AB resulted in improved BWG. These results suggests that a right dosage (30 g/kg of feed) of EM is required to exhibit growth stimulation effects in broilers. At this inclusion level, addition of AB produces a complimentary effect. There were no differences in the FCR reported in this study. However, the FCR tended to improve with addition of EM or AB which may be a reason for the improvements in BWG. Increased BWG was also associated with high FI. Mohan et al (1996), attributed the growth promotion effects of probiotics to improvements in utilisation of apparent metabolisable energy while enhanced feed utilisation was mentioned by Jin et al (1998).
The high dressing percentage for birds fed the 30 g/kg EM supplemented diet was concomitant with the high BWG observed in the respective treatments. Other studies have shown no differences in dressing percentage between probiotic and no- probiotic supplemented diets (Mohan et al, 1996).
In agreement with other studies involving broilers (Mohan et al, 1996; Jin et al, 1998), this study has demonstrated that probiotics such as EM have serum CHOL reducing properties. These effects are dose dependent and according to this study, the appropriate dosage is 30 g/kg. Eyssen (1973) reported that deconjugation of bile acids in the small intestine may be responsible for reduction of concentrations of serum cholesterol because deconjugated bile acids do not function as well as conjugated bile acids in solubilisation and absorption of lipids. Chikai et al (1987) reported that adherence of deconjugated free bile acids to bacteria and dietary fiber enhances excretion of the bile acids. This mechanism has been implicated to trigger a feedback mechanism that regulates hepatic cholesterol synthesis and subsequent transformation into bile acids which may be responsible for lowering serum cholesterol levels.
A mortality rate of only 0.22 per cent was recorded for the whole trial. Although a prophylactic effect by probiotics has been reported in other studies, (Cavazzoni et al, 1998), it is difficult to draw a similar conclusion in the study presented here because there are no differences in the morbidity and mortality cases among the six treatments some of which had no EM added.
In conclusion, this study has demonstrated and substantiated earlier reports that probiotics such as Effective Microorganisms (EM) has growth promoting effects. Additionally, supplementation of broiler diets with EM has serum cholesterol reducing properties, an element important for the health conscious consumers. A dosage of 30 g/kg is required if the aforementioned benefits are to be attained.
Finally, although further research on beneficial effects of dietary EM is required, the results reported herein bear testimony to the fact that probiotics such as EM offer a potential alternative to antibiotics in broiler diets.
References:
Cavazzoni V., A. Adami, and C. Castrovilli (1998). Performance of broiler chickens supplemented with Bacillus coagulans as probiotic. British Poultry Science 39: 526-529.
Chikai T., H. Naka and K. Ushida. (1987). Deconjugation of bile acids by human intestinal bacteria implanted in germ-free rats. Lipids 22: 669
Eyssen, H. (1973). Role of gut microflora in metabolism of lipids and sterols. Proceedings of the Nutritional Society 32:59
Jin L.Z., Y.W. Ho, N. Abdullah and S. Jalaludin (1998). Growth performance, intestinal populations and serum cholesterol of broilers fed diets containing Lactobacillus cultures. Poultry Science 77: 1259-1265.
Mohan B., R. Kadirvel, A. Natarajan and M. Bhaskaran (1996). Effect of probiotic supplementation on growth, nitrogen utilisation and serum cholesterol in broilers. British Poultry Science 37: 395-401
Phillips J.M. and S.R.Phillips (1996). The APNAN User’s Manual: EM Nature Farming Guide. First U. S. Edition. EM Technologies, Inc., Tucson, Arizona, USA.
SAS® Institute (1988). SAS/STAT User’s Guide, Release 6.03 Edition. SAS Institute Inc. Cary, NC.
Acknowledgements
Financial Assistance was provided by Bunda/SACCAR GTZ Animal Science Programme (University of Malawi). Mr Yoshida of EMROSA (Pty) Ltd., South Africa, donated the EM used in this study. Their contributions are highly appreciated.
Immunomodulatory Effect of Effective Microorganisms® (EM®) in Chickens
Wondmeneh Esatu, Getachew Terefe and Tadelle Dessie
ABSTRACT:
The study was conducted to determine the immunomodulatory effects of Effective Microorganisms (EM ®) on Horro and Fayoumi chickens at Agricultural Research Center and National Veterinary Institute (Debre Zeit). A total of 450 chickens (225 from each breed) were used in this study. Birds were grouped according to treatment: EM-treated (with feed, with water, with feed and water) and non-treated (only SRBC treated and Non-EM, Non-SRBC) controls. EM was given daily from the 3rd week of age for 5 weeks. Birds were injected with SRBC on the 4th and 6th week to see the total, IgG and IgM antibody responses. Antibodymeasurements were made using hemagglutination technique. The findings show that: (1) EM application has significantly increased antibody responses to SRBC, (2) there was no difference in antibody responses between the two breeds, or between the three modes of EM application. From these, it can be concluded that EM has a positive immunomodulatory effect when provided to chicks with feed or water. This study did not consider the cellular arm of the immune response and EM response to infections with specific pathogens was not investigated, collectively demanding further research in these and other issues if EM has to be used as good feed additive.
How to cite this article: Wondmeneh Esatu, Getachew Terefe and Tadelle Dessie, 2012. Immunomodulatory Effect of Effective Microorganisms (EM®) in Chickens. Research Journal of Immunology, 5: 17-23. DOI: 10.3923/rji.2012.17.23 URL: http://scialert.net/abstract/?doi=rji.2012.17.23 Received: October 30, 2011; Accepted: November 14, 2011; Published: June 19, 2012.
INTRODUCTION
Available experimental data show that our indigenous birds have limited genetic capacity for both egg and meat production ( Negussie, 1999). However, local chickens have several invaluable characteristics appropriate to traditional low input/low output farming systems, which are not found in any exotic breed ( Tadelle, 2003). Horro is one of local chicken known for its reasonably good performance among ecotypes studied in Ethiopia ( Tadelle, 2003). The problem of poor adaptability to confinement and susceptibility of the Horro ecotype to some infectious diseases compared to the Fayoumi breed has prompted the DZARC to undertake a comparative study on their immune responses using Effective microorganisms, a laboratory cultured mixture of microorganisms consisting mainly of lactic acid bacteria, photosynthetic bacteria and yeast. Studies in other areas have already shown that the use of probiotics and Effective microorganisms could improve the immune responses of chicken to various infections and enhance better sanitation in poultry houses ( Kabir et al., 2004; Mohiti et al., 2007; Chichlowski et al., 2007). The objective of this study was to evaluate the immune competence (Humoral immune response) of Horro chicks compared with Fayoumi breed with or without EM supplementation.
MATERIALS AND METHODS
Study animals, Housing and feeding of experimental chicks: Day old chickens obtained from Fayoumi and Horro chickens were randomly assigned to experimental pens and reared in floor pens filled with hay as litter material with a density of 6 birds m -2. A standard starter feed and water ad libitum till 8 weeks of age (end of experimental study). Standard bio-security protocol was employed throughout the experimental period; however, the chicks were not vaccinated for any of the prevalent diseases.
Experimental design and treatment groups: Experimental groups of Fayoumi and Horro chickens with different modes of EM treatment were arranged in a Completely Randomized Design (CRD) with three replications of 15 birds each ( Table 1).
Supplementation of Effective Microorganisms (EM®): The different EM preparations were made according to EM research organization ( EMROSA, 2003 ). Birds were provided with EM with feed, with water or with feed and water daily starting from the age of 3 weeks till the end of the experiment (week 8). For this, 1% EM-Bokashi in feed was made for EM with feed, 0.1% EM-activated solution in water for EM in water and half of the above concentrations for EM supplemented in both feed and water. Control groups (F-NT-C, F-SRBC, H-NT-C and H-SRBC) did not receive EM.
Table 1: Experimental groups of Fayoumi and Horro chickens with different modes of EM treatment
F-EM-F: Fayoumi given EM in feed (*3), F-EM-W: Fayoumi given EM in water (*3), F-EM-FW: Fayoumi given EM in Both feed and water (*3), F-SRBC: Fayoumi SRBC treated control (*3) F-NT-C: Fayoumi non-treated control (*3), H-EM-F: Horro given EM in feed (*3), H-EM-W: Horro given EM in water (*3), H-EM-FW: Horro given EM in Both feed and water (*3), H-NT-C: Horro non-treated control (*3), H-SRBC: Horro SRBC treated control (*3)
Measurement of immune response
Preparation of SRBC and immunization of chickens: Sheep red blood cells were isolated and a 0.5% suspension of the SRBC was made with PBS at the National Veterinary Institute (Debre Zeit, Ethiopia) using a standard technique. All groups of birds except controls were then immunized intramuscularly (in the breast area) with an initial dose of 0.5 mL of 0.5% SRBC in PBS one week after EM supplementation was started (week 4). Negative control groups received the same amount of PBS in place of SRBC. The booster dose of SRBC was given on week 6, i.e., 14 day after the initial injection of the SRBC antigen.
Measurement of antibody responses against SRBC: Serum samples were collected from brachial vein weekly starting from the age of 3 weeks. The first sample (week 3) was meant to represent antibody response against SRBC before the start of any treatment (EM or SRBC). The second sample represents antibody response against SRBC after one week of EM supplementation but without SRBC injection. Measurements were made for total and IgG and IgM antibodies according to the method described previously ( Yamamoto and Glick, 1982; Qureshi and Havenstein, 1994; Lepage et al., 1996).
Statistical analysis and model: Serum Ig titers (Ig-total, IgG, IgM) were monitored in a 2*5 ANOVA (two strains*4 EM supplementations and additional two SRBC subjected but non EM injected controls) . All data were analyzed by ANOVA with the repeated model mixed procedure of SAS software (2000 version 8) and compared by least square means at p<0.05.
RESULTS
Antibody responses to EM supplementation (Total, IgG and IgM)
Total immunoglobulin titer: Antibody titers for samples taken before any treatment or before SRBC treatment were much lower compared to those after treatment with EM. EM-treated groups had significantly higher total immunoglobulin titers compared to the non-treated control groups for each breed (p<0.05). The peaks were observed 3 weeks after the start of EM and 2 weeks after the first SRBC injection. However, it gradually declined till week 8 even though it remained significantly higher than control groups ( Fig. 1). There was no significant difference for total antibody titer between EM treatment groups in each breed and between breeds for each mode of EM application (feed, water, both). One exception to this is, the significantly higher antibody titer in Fayoumi breed compared to Horro when EM is given with water (p<0.05). On the other hand, total antibody titer in groups treated with SRBC where significantly higher than the non treated control groups in both breeds during the study period (p<0.05). As the total antibody titer was very low before SRBC treatment, it was difficult to obtain readings for the fractions (IgG and IgM). Therefore, data for weeks 3 and 4 are not included here. Starting from the 5th week of age (2 weeks of EM and 1 week of SRBC), the pattern of IgG titer is similar to that of the total antibody measurement ( Fig. 2). All EM supplemented groups had higher IgG titer than control and SRBC groups for both breeds (p<0.05) except on week 5 where there was no significant difference between EM-treated and non-treated groups. There was no significant difference for IgG antibody titer between EM treatment groups in each breed and between breeds for each mode of EM application. However, on week 8 there was significantly higher IgG titer in Fayoumi breed than in Horro when EM is given with water (p<0.05). IgM response peaked one week following the first SRBC injection and then declined to control levels within the following two weeks ( Fig. 3 ).
Fig. 1: Total antibody titre of Fayoumi (F) and Horro (H) chicken groups supplemented with EM in feed (EM-F), in water (EM-W), in feed and water (EM-FW) and non-treated control (C) and SRBC groups
Fig. 2: Dynamics of IgG antibody in Fayoumi (F) and Horro (H) chicken groups supplemented with EM in feed (EM-F), in water (EM-W), in feed and water (EM-FW), non-treated control (C) and SRBC groups
Fig. 3: Dynamics of IgM antibody in Fayoumi (F) and Horro (H) chicken groups supplemented with EM in feed (EM-F), in water (EM-W), in feed and water (EM-FW), non-treated control (C) and SRBC groups
EM application, regardless of the mode of application, has resulted in significantly higher IgM titer on weeks 5 and 6 compared with controls in both breeds of chicken (p<0.05). For those groups treated with EM, there was no significant difference in IgM titer between the different modes of EM supplementation and between the two breeds.
DISCUSSION
Effect of EM on immune competence of horro and fayoumi chickens: The significance of EM/probiotic supplementation in boosting immune response to various antigens such as SRBC and specific pathogens has been demonstrated in various studies. Anjum (1998) has reported that geometric mean titer of antibody against New Castle Disease in EM treated chickens was twofold higher compared with untreated control layers. Similarly, Kabir et al. (2004), Perdigon et al. (1995), Panda et al. (2000), Cross (2002) , Yunis et al. (2000), Koenen et al. (2004) and Huang et al. (2004) reported that probiotics stimulate the immunity of chickens when compared with non-probiotic groups. In this study, EM supplementation irrespective of the mode of application had a positive effect on total antibody as well as IgG and IgM levels in both Fayoumi and Horro chicken. This is clearly visualized when mean agglutination titers are compared to those of non-EM controls. A previous study demonstrated that day-old chicks immunized with probiotics had increased serum and intestinal antibodies reactive to tetanus toxoid and Clostridium perfengens alpha toxin (Haghighi et al., 2006). This result is also in line with the findings of Ahmad (2006) who reported EM increases production of antibodies usually of IgG and IGM classes and interferon γ. Similarly, ( Haghighi et al., 2005 ) also reported that Probiotic treated birds showed significantly higher serum antibody to SRBC than birds that were not treated with probioties. Likewise, serum IgG and IgM reactive to tetanus toxoid and alpha toxin were increased in probiotic treated, unimmunized chickens compared to levels in untreated controls ( Haghighi et al., 2006). The sharp increase in the level of IgM in all EM-treated groups one week after SRBC injection observed in this study is in accordance with other studies. IgM appears after 4-5 days following exposure to a disease organism/antigen and then disappear by 10-12 days ( Butcher and Miles, 2003).The mechanisms by which EM/probiotics improve the immune responses of birds are not clearly understood. The addition of probiotics to diets may benefit the host animal by stimulating appetite, improving intestinal microbial balance and stimulating non-specific/specific immune system ( Nahashon et al., 1992; Afrc, 1989 ; Toms and Powrie, 2001). The present study compared the Fayoumi breed (originated from Egypt) known for its adaptability to various environmental conditions and its relative resistance to some known diseases ( Pinard-Van Der Laan et al., 1998; Lakshmanan et al., 1996) with the poorly known local chicken, the Horro. The findings indicated that except at week 8 where the Fayoumi chicken responded better than the Horro when EM is given with water or with feed and water, there was no significant difference between the two breeds for total and specific antibody responses against SRBC irrespective of the type of EM supplementation. It seems that both the local Horro and the Exotic Fayoumi have similarly responded to EM supplementation. The exceptions at week 8 could be due to the amount of EM-treated water consumed. Similarly, Deif et al. (2007) have reported that Hubbard broiler chicks had significantly higher total ant-SRBC antibody titer at 7 and 14 days post primary and secondary SRBC injection compared to Cobb breed. Such discrepancy in results may arise due to differences in experimental animals, experimental protocols and/or study duration (5 weeks in our case). It was observed that mode of application had generally no remarkable influence on the immunomodulatory effect of EM in both Fayoumi and Horro chicken. This may imply that mixing the EM activated solution in water or feed (as medium of administration) did not significantly affect the immunomodulatory activity of EM.
CONCLUSION AND RECOMMENDATION
EM treatment caused a significant increase in total, IgG and IgM antibody levels (against SRBC as a general mitogen) compared to non-treated controls in both breeds. These responses were not different between breeds and between modes of EM application. Therefore, EM can be used as a feed additive regardless of the mode of application (feed/water) to boost the immune responses of these birds. However, it remains to be seen if the addition of EM to feed or water could also improve the immune competence of these birds to specific pathogens. However, this study was conducted with limited number of chicks and with relatively young birds (less than two months of age) due to limitations in availability of reagents. Therefore, to arrive at better understanding and conclusion on the role of EM and its future utilization in poultry farming. It is essential to enhance resistance of chicks to the most important diseases such as viral and bacterial origin. therefore, experimental works should involve the role of EM to boosting immune responses to specific pathogens.
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Ahmad, I., 2006. Effect of probiotics on broilers performance. Int. J. Poult. Sci., 5: 593-597.
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Lepage, K.T., S.E. Bloom and R.L. Jr. Taylor, 1996. Antibody response to sheep red blood cells in a major histocompatibility (B) complex aneuploid line of chickens. Poult. Sci., 75: 346-350.
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Negussie, D., 1999. Evaluation of the performance of local, Rod Island Red and Fayuomi breeds chicken under different management regimes in the highlands of Ethiopia. M.Sc. Thesis, Swedish University of Agriculture Sciences, Uppsala.
Panda, A.K., M.R. Reddy, S.V.R. Rao, M.V.L.N. Raju and N.K. Praharaj, 2000. Growth, carcass characteristics, immunocompetence and response to Escherichia coli of broilers fed diets with various levels of probiotic. Arch. Geflugelkd., 64: 152-156.
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Pinard-Van Der Laan, M.H., J.L. Monvoisin, P. Pery, N. Hamet and M. Thomas, 1998. Comparison of outbreed lines of chickens for resistance to experimental infection with Coccidiosis (Eimeria tenella). Poult. Sci., 77: 185-191.
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Influence of Effective Microorganisms® on Health and Immune System of Broilers Under Experimental Conditions
Ahmed D Anjum, Tahir Hussain, Farzana Rizvi, Ghulam Gilani and Tariq Javaid University of Agriculture, Faisalabad-38040, Pakistan.
Abstract
Eighty, day-old, commercial broilers were divided in four equal groups and were fed standard broiler ration. Effective microorganisms (EM) were administered in solution form through drinking water to 1st group, in solid form (Biofeed) through feed to 2nd group and both to 3rd group from 10 days of age to 56 days of age. The 4th group served as control. Birds were vaccinated with Newcastle disease (ND) vaccine at 7 days and 28 days of age.
After 45 days of EM treatment live body weight was 2004=F1 45.1 g, 1978 =F1 61.5 g and 2022 =F1 45.4 g, respectively, in broilers given EM solution alone, biofeed alone and both simultaneously compared with 1690=F1 37.1 g in the control broilers (P<0.001 ). Of the lymphoid organs bursa and thymus indices were significantly greater in broilers given EM solution alone and both EM solution and biofeed compared with the control broilers (P<0.05) at 45 days post-treatment. Spleen weight was not influenced with EM treatment.
Of the visceral organs liver index was significantly lesser in broilers given biofeed alone compared with the control broilers (P<0.01). Gizzard index was significantly lesser in birds given EM solution (P<0.05), biofeed (P<0.001) and both (P<0.01) than the control broilers. Heart index was significantly lesser in birds given EM solution (P<0.01), biofeed (P<0.01) and both (P<0.05) than the control broilers.
Proventriculus, intestine, kidneys, pancreas and thyroid Indices did not differ significantly between EM treated and control broilers at 45 days post-treatment. Antibody geometric mean titre (GMT) value against ND vaccine virus during primary response was 13 in broilers given EM solution alone, 9.8 in broilers given biofeed alone and 9.8 in broilers given both simultaneously compared with 3.2 in the control broilers. The counterpart GMT values during secondary response were 445.7, 264, 256 and 68.6.
In conclusion, EM is a safe product. This technology can be applied for promoting growth and for potentiation of immune response in the chicken.
Introduction
Effective microorganisms (EM) is a newer technology from Japan recently introduced in Pakistan by Hussain et al., 1996. EM is available in the form of solution and solid for commercial use. Until now the products are being used mainly in Agriculture and the technology has shown promising results in plant health and production. There were reports from abroad on EM use in livestock and poultry. This study was envisaged to investigate the effect of EM on health and immune system of poultry under local conditions.
Materials and Methods
Experimental birds Eighty day-old, commercial broiler chicks of certified health status were procured from a breeder company (Mughal Chicks Sargodha). The chicks were divided into four equal groups and kept in cages under standard management conditions. The chicks were fed adlibitum commercial broiler ration (Punjab Feeds Ltd.) containing 23 per cent crude protein and 3000 ME/kg throughout the study period of eight weeks.
Effective Microorganisms
EM was supplied by Hussain et al. (1996) in solution and solid (biofeed) form. EM primarily contains Lactic acid bacteria and other useful bacteria. EM was given as solution at the dose rate of 1 ml/L drinking water to first group, as biofeed at the dose rate of 30 g/Kg in feed to second group and both simultaneously to third group continuously from 10 days of age to 56 days of age. The fourth group served as control.
Parameters
The following parameters were observed.
1. Weekly live body weight.
2. Feed consumption and feed conversion ratio.
3. Organ weights at four weeks and eight weeks of age.
4. Antibody titre after primary and booster vaccination with Newcastle disease vaccine virus (LaSota strain, Bioteke Italy).
Results
Live body weight
Live body weight in experimental broilers is given in Table 1. It was significantly greater in broilers given EM solution alone or biofeed alone than the control from 4th week post- treatment onward (P<0.001). Whereas live body weight was significantly greater in broilers given both EM solution and biofeed simultaneously than the control broilers from 3rd week post-treatment onward (P<0.001). At the end of 45 days of treatment live body weight was 18.6 percent, 17 percent and 19.6 percent greater, respectively, in broilers given EM solution alone, biofeed alone and both simultaneously than the control broilers. Net weight gain in the three respective treatments was 1912 g, 1884 g and 1919 g compared with 1596 g in the control broilers.
Feed conversion ratio
Feed consumption was 5.11 kg in broilers given EM solution, 5.12 kg in broilers given biofeed and 5.18 kg in broilers given both simultaneously compared with 4.93 kg in the control. Feed conversion ratio was 2.55 in broilers given EM solution, 2.59 in broilers given biofeed and 2.56 in broilers given both simultaneously compared with 2.92 in the control.
Lymphoid organs
Actual weight of bursa and thymus and their indices were significantly greater in EM treated broilers compared with the control (Table 2, P<0.05). Actual weight of spleen was significantly greater in EM treated broilers compared with the control (P<0.05) but its index did not differ significantly between EM treated broilers and the control (Table 2).
Visceral organs
Table 3 shows that actual weight of liver was significantly greater in EM treated broilers compared with the control but liver index was significantly lesser in EM treated broilers compared with the control (P<0.05). Actual weight of proventriculus was significantly greater in EM treated broilers compared with the control (P<0.05) but its index did not differ significantly between EM treated broilers and the control. Actual weight of gizzard did not differ significantly between EM treated broilers and the control but gizzard index was significantly lesser in EM treated broilers compared with the control (P<0.05). Actual weight of intestine and its Index did not differ significantly between EM treated broilers and the control. Actual length of intestine was significantly lesser in EM treated broilers compared with the control (P<0.01). Actual weight of pancreas was significantly greater in EM treated broilers compared with the control (P<0.05). Pancreas index did not differ significantly between EM treated broilers and the control. Actual weight of kidney and its index did not differ significantly between EM treated broilers and the comrol. Actual weight of heart did not differ significantly between EM treated broilers and the control. Heart index was significantly lesser in birds given EM solution (P<0.01), biofeed (P<0.01) and both (P<0.05) than the control broilers. Actual weight of thyroid and its index did not differ significantly between EM treated and control broilers at 45 days post-treatment.
Antibody titre
Primary and secondary immune responses, in terms of antibody geometric mean titre (GMT) were determined against Newcastle Disease (ND) vaccine virus. GMT values during primary response were 13 in broilers given EM solution alone, 9.8 in broilers given biofeed alone and 9.8 in broilers given both simultaneously compared with 3.2 in the control broilers. The counterpart GMT values during secondary response were 445.7, 264, 256 and 68.6.
Discussion
Effective micro-organisms were administered to broiler chicks in the form of “EM solution” “biofeed” and “both simultaneously” to study their effect on health and immune response of broiler chicks. Live body weight was significantly greater in all EM treated groups compared with the control (Table 1, P<0.001). It confirms growth promoting activity of EM as reported by other workers (Hussien and El-Ashry, I991; Ahmad, 1996; Hussain 1996). The better weight gain in EM-treated broilers could be related to better digestibility of crude protein and crude fiber (Hussain, 1996). Interestingly, the increase in live body weight was also accompanied with a significant decrease in offal weight i.e., of liver index (P<0.05), gizzard index (P<0.01), intestinal weight index, intestinal length index (P<0.05). kidneys index (P<0.05) and heart index (P<0.01) (Table 3). In Pakistan there are numerous antibiotics widely used as growth promoters but all have associated risks of drug residues and drug resistance. The EM appears to be a safe growth promoter without any associated risks.
Of the visceral organs the decrease in heart index is of particular interest and needs further consideration. This has also been observed in a previous experiment (Ahmad, 1996). Heart enlargement is a common disease in humans. If constituents of the EM responsible for heart enlargement can be determined and purified, further research may indicate if this can help in treatment of heart enlargement.
The greater bursa and thymus index in birds supplemented EM as compared to the control (Table 2, P<0.05) suggests that EM supported these lymphoid organs. The two lymphoid organs are responsible for recruiting B and T Iymphocytes and make up the vital and basic components of humoral and cellular immunity. The gross support of lymphoid organs was also accompanied with better antibody production in EM treated broilers. In the present study GMT against Newcastle disease vaccine virus was 6.5 times in broilers given EM solution, 3.85 times in broilers given biofeed and 3.73 times in broilers given both EM solution and biofeed simultaneously than the control value.
The present study showed that EM potentiated immune response in the experimental broilers. Previous studies demonstrated that Lactic acid bacteria administered orally or intraperitoneally enhanced activity of the mononuclear phagocytic system (Kato et al., 1983) and increased the production of circulating antibodies for certain antigens in mouse (Saito et al., 1983; Perdigon and Alvare, 1996). However, further investigation is required for the elucidation of the mechanism through which EM produced systemic increase in the immune response.
References
Ahmad,A. 1996. Personal communication
Hussain, I., 1996. Effect of microbial culture (EM4) on the performance of male broiler chicks. M.Sc. thesis, Department of Poultry Husbandry, UAF, Pakistan.
Hussain, T., R. Ahmad, G. Gilani and T. Higa, 1996. Applied EM Technology, Nature Farming Research Centre, University of Agriculture Faisalabad, Pakistan.
Hussein, H.H. and M.A. EI-Ashry, 1991. Some studies on the beneficial effect of Lactobacillus concentrate supplementation on broiler performance. Egyptian Journal of Animal Reproduction 28(1):85-91.
Kato, I., T. Yokokura and M. Mutai, 1983. Macrophage activation by Lactobacillus casei in mice. Microbial. Immunol., 27(7):611-618.
Perdigon, G. and S. Alvarez, 1995. Probiotics and the immune state. In: Probiolics. Ed. R. Fuller. pp.1-25.
Saito I. K. Sato and Y. Horikawa 1983. Enhanced humoral antibody production and delayed type hvpersensitivity response in mice by Lactobacillus casei. Hiroshima J. Med. Sci., 32:223- 225.
Table 1. Effect of effective microorganisms on live body weight (grams) of experimental broilers.
Each figure represents mean (=F1 standard error of the mean) of 10 birds. ***P<0.001 compared with the control.
Table 2. Actual weight and index of lymphoid organs in experimental broilers at 45 days post EM treatment.
Each figure represents mean (=F1 standard error of the mean) of 10 birds.
*P<0.05, **P<0.01, ***P<0.001 compared with the control.
Table 3. Actual weight and index of visceral organs in experimental broilers.
Each figure represents mean (=F1 standard error of the mean) of 10 birds.
*P<0.05, **P<0.01, ***P<0.001 compared with the control group. Age (week)
Influence of Effective Microorganisms® on the Quality of Poultry Products
S. Chantsavang1 and P. Watcharangkul2
Department of Animal Science, Kasetsart University, Bangkok, Thailand1 and
Nutrition Division, Department of Health, Ministry of Public Health, Nonthaburi, Thailand2
Abstract
Experimental results from 4 different types of poultry were summarized to reveal the application of EM for production of quality and healthy animal products. In low production animal as Muscovy duck, a battery cage trial was conducted. Results of 16 week experiment showed that EM added in feed and/or in drinking water had no significant effect on production performance but significantly increased breast percentage, significantly reduced breast ash content and tended to increase protein content of breast meat and polyunsaturated fatty acids content in duck oil.
In high production animal, Arbor Acres broiler chickens were used to evaluate the effect of EM. Results of 7 weeks experiment showed that EM added in feed and/or in drinking water had no effect on production and carcass characteristics but tended to decrease ash content of breast meat. Other quality traits included in the study were fatty acid and cholesterol composition, E. coli and Salmonella contamination. In laying chickens, EM added in feed had no significant influence on all traits studied except for such an important quality trait as yolk color. Layers received EM in feed gave darker color yolk. Similar results were also obtained in a 12 week experiments in Japanese quail.
The Effect of Effective Microorganisms® on Production and Quality Performance of Rhode Island Red layers
International Journal of Livestock Production Vol. 4(2), pp. 22-29, February 2013 Available online at http://www.academicjournals.org/IJLP
DOI: 10.5897/IJLP12.015
ISSN 2141-2448 ©2013 Academic Journals
Full Length Research Paper
M. Simeamelak1*, D. Solomon2 and T. Taye2 1Southern Institute of Agricultural Research, Hawassa Agricultural Research Center, Ethiopia.
2Jimma University, College of Agriculture and Veterinary Medicine, Jimma, P. O. Box 307, Ethiopia.
Accepted 10 September, 2012
Abstract
Rhode Island Red (RIR) breed of chickens are reported to be capable of acclimatization to the Ethiopian rural production environment. However, there have been serious complaints that the reproduction performance of RIR breeds of chicken is low. This study was conducted to evaluate the effect of Effective Micro-organisms (EM) on reproduction performance of Rhode Island Red (RIR) layers. A total of 96 RIR pullets of 16 weeks old were divided into 8 groups, each with 12 pullets. These were randomly assigned to 4 treatments containing 0, 4, 8 and 12 ml of EM/liter of drinking water in completely randomized design with 2 replications for a study period of 22 weeks. Feed consumption, feed conversion efficiency, egg production, egg quality, fertility, and hatchability were used as evaluation parameters. The results obtained showed that there was no significant difference among all the treatment groups in feed consumption, sexual maturity, survival rate and feed conversion efficiency (P>0.05) to an age of pullets, whereas the mean body weight gain of the groups of 24 weeks placed on the treatment containing 8 to 12 ml of EM/liter of drinking water were significantly (P<0.05) higher than the control groups. The results obtained also showed that there was no significant (P>0.05) difference between all the treatment groups of layers in feed consumption, fertility and hatchability (P<0.05) to an age of 37 weeks. On the other side, the mean weekly egg production and feed conversion efficiency during the laying period were significantly higher (P<0.05) for the groups of layers placed on the treatment containing 4 to 12 ml of EM/liter of drinking water compared to that of the groups placed on the control treatments. In summary, the results of this study showed that inclusion of 4 to 12 ml of EM/liter of drinking water resulted in significant improvement in survival and growth rate, egg production, feed conversion efficiency and egg quality parameters. Extending EM technology to indigenous chickens could be the future direction of research.
Key words: Egg production and egg quality, effective micro-organisms, feed conversion, Rhode Island Red (RIR) chickens.
INTRODUCTION
The introduction of exotic chickens into Ethiopia dates back to the early 1950's, when Rhode Island Red (RIR) breed of chickens were imported along with other exotic genetic materials. It was the Ministry of Agriculture (MoA) that was given the mandate for national poultry extension work from the very beginning, and MoA established several poultry breeding and multiplication centers in different parts of the country. The centers were involved in the distribution of fertile eggs, day old chicks, pullets/cockerels, culled layers and provision of management information of Rhode Island Red (RIR) breeds of chickens to the rural farming population. The RIR breed of chickens distributed were reported to be capable of well acclimatization to the Ethiopian rural production environment with reasonable production level under smallholder management systems. However, there have been serious complaints by the farming community and the multiplication centers, suggesting that the production performance of RIR breeds of chickens is low as measured by age at sexual maturity, rate of egg production, fertility and hatchability. The information obtained from Amhara Regional State, Rural Development Bureau of Agriculture indicates that the farming community is facing problems as a result of poor fertility and hatchability of the RIR breed of chicken distributed. It was also reported that there is improvement in the production and reproduction performance of poultry with the addition of Effective Micro-organism (EM) (Safalaoh and Smith, 2001).
Effective Micro-organisms are live microbial feed supplements with beneficial effect to the host animal by improving its intestinal microbial balance (Fuller, 1989). A diverse micro-biota was found throughout the digestive tract of animals with relatively higher concentration in the cecum (Mead, 1997). This micro flora has a role in nutrition particularly in the area of detoxification of certain compounds, stimulation of animal growth, and improvement of the health status and well-being of the host animals through protection against pathogenic bacteria (Van der Wielen et al., 2002). The improvement in production performance of poultry fed on the ration containing EM was reported to be attributed to the improvement in feed bioavailability, balance of gastrointestinal micro-organisms, and enhancement of the immunity status of the birds. EM was reported to be successfully used for increasing productivity in integrated animal units and poultry farms in South Africa (Hanekon et al., 2001; Safalaoh and Smith, 2001). Effective Micro- organism has also been used to improve growth and egg production performance of poultry (Stavric and Kornegay, 1995).
Inclusion of EM dominated by Lactobacillus acidophilus in laying hens diets was reported to have improved some quantitative and qualitative parameters of eggs. There has been an increase in the number of laid eggs, decrease in feed intake, improvement in feed conversion ratio, egg specific gravity and an increase in the Haugh Units (Daniele et al., 2008). Panda et al. (2003, 2008) reported significant increase in the egg production performance of White leghorn layers with dietary supplementation of a probiotic (L. sporogenes) at the rate
Table 1. Treatment Allocation to the experimental birds.
of 100 mgkg−1 diet (6 × 108 spores). All these probiotics and experimental EM effects showed that the use of standardized EM would have improvement effect on layers. Therefore, the objective of this study was to solve the problem of reproduction performance of RIR layers.
METHODOLOGY
Description of experimental site
This experiment was conducted at Jimma University College of Agriculture and Veterinary Medicine (JUCAVM), located at 357 km southwest of Addis Ababa at an altitude of 1710 m above sea level. The mean maximum and minimum temperature of the study area was 26.8 and 11.4°C, respectively and the mean maximum and minimum relative humidity was 91.4 and 39.92% respectively. The mean annual rainfall of the area is 1500 mm (BPEDORS, 2000).
Experimental treatments
Adequate quantities of activated EM·1® packed in plastic jar was obtained from Weljijie PLC located in Debre Zeit which intern located at 70 km east of Addis Ababa. Weljijie PLC obtains the original EM·1® culture from EMRO Malaysia Sdn. Bhd. Activated EM·1® was made at the ratio of 5% molasses, 5% original EM-1® of the total volume which was mixed with chlorine free clean water. The major groups of micro-organisms in EM-1® are lactic acid bacteria, yeast and phototrophic bacteria. Activated EM·1® was transported to JUCAVM poultry farm and stored properly until required for the formulation of the experimental treatments. Four experimental treatments shown in Table 1 were prepared by inclusion of 0, 4, 8 and 12 ml of EM solution/liter of chlorine free drinking water. The treatments were prepared on daily basis.
Management of the experimental birds
A total of 100 RIR pullets at an age of 12 weeks were purchased from Southern Nation Nationality and peoples State poultry breeding and multiplication centre located in Bonga and transported to JUCAVM poultry farm. These were housed in well prepared grower’s house and placed on grower’s commercial ration. At 16 weeks of age, 96 pullets were divided into 8 groups, each with 12 pullets. Two cockerels of the same age and breed were assigned to each group and each group was housed in separate pens of equal dimension that were properly cleaned, disinfected, and provided with all the necessary layers house equipments in advance. Finally, the 4 treatments were randomly assigned to the experimental pullets with two replications (smaller replication was due to shortage of experimental house during the study period) for the study period of 22 weeks (Table 1). At 5 months of age, all the treatment groups were switched to commercial layers ration; the feed composition is a secret of the factory, quality feed manufacturing factory. All the treatment groups were fed to appetite and chlorine free water containing different levels of EM (treatments) was made available at all times.
Table 2. Weekly mean feed consumption (g/head) of pullets placed on different levels of EM.
s. e = standard-error; Means in a row without superscripts are statistically not significant (p>0.05); T1 = control; T2 = 4 ml of EM/liter of water; T3 = 8 ml of EM/liter of water; T4 = 12 ml of EM/liter of water.
Egg quality determination
Twelve eggs laid during the last three consecutive days of the 7 weeks laying period were randomly selected from each treatment. The eggs were individually weighed, carefully opened (broken) onto a flat plate and the yolk and albumen were separately weighed. Yolk height was measured using tripod micrometer (0.01 mm gauge) and yolk index was calculated according to the method described by Akhtrs (2007). Egg shell thickness was measured using calibrated micrometer screw gauge. Yolk color was measured using roach color fan. Haugh unit was calculated using the formula adopted from the study of Haugh (1937).
Fertility and hatchability determination
Fifty fresh eggs (stored for 10 days) were taken from each treatment, selected against undesirable shape, size and shell structure and incubated. The eggs, incubator and all the fixtures were fumigated with formalin plus potassium permanganate (Altman et al., 1997). The incubation temperature, humidity and turning device were adjusted in advance according to the recommendations of the manufacturer. Candling was done on the 7th and 14th day of incubation aimed at calculating fertility and hatchability.
Statistical analysis
Since repeated data were collected on the same animal daily/weekly it was appropriate to use Repeated Measures Design (RMD). Data on body weight gain, feed consumption, feed conversion ratio, sexual maturity, and rate of egg production, egg quality, fertility and hatchability were collected throughout the study period. The data collected were subjected to Repeated Measures Design (RMD) of SAS 9.00 version for analysis (SAS Institute, 2002). Least square mean were used for comparison.
RESULTS AND DISCUSSION
Feed consumption during growing
There was no significant (p>0.05) difference between all the treatment groups in mean weekly feed consumption to an age of 24 weeks, though the groups receiving 0 ml of EM/liter of drinking water tended to consume more than the others (Table 2). The other treatment groups showed proportional reduction in feed consumption as a result of increase in the volume of EM administered /liter of drinking water.
Similarly there was no significant difference between (P>0.05) all the treatment groups in weekly body weight gain during the first 5 weeks of the feeding trial. Weekly body weight gain brought by the treatment groups assigned to the control treatment was significantly (P<0.05) lower than the groups placed on the treatment containing 8 to 12 ml of EM/liter of drinking water during the last 4 weeks of feeding. There was no significant difference between the treatment groups assigned to 4 to 12 ml of EM/liter of drinking water in weekly body weight gain and feed conversion efficiency at any time of the feeding trial (Tables 3 and 4).
Significant (P<0.05) difference in mean daily body weight gain between the treatment groups of pullets was recorded after 5 weeks of the feeding trial whereas: there was no significant difference in feed consumption between all the treatment groups at any time The results of this study are in agreement with that of Kalavathy et al. (2003) who reported improved body weight gain of broiler with supplementary administration of Lactobacillus. Mean weekly feed consumption of T1, T2, T3 and T4 833.89, 793.85, 766.07 and 754.36 g head was attained by the groups placed on 0, 4, 8 and 12 ml of EM/liter of drinking water respectively (Table 5). Similar trend was also reported by Balevi et al. (2009) from the trial conducted to study the effect of dietary supplementation of commercial
Table 3. Mean weekly body weight gain (g/head) of pullets placed on different level of EM.
s. e = standard-error; Means in a row having similar superscripts are statistically not significant (p>0.05); T1 = control; T2 = 4 ml of EM/liter of water; T3 = 8 ml of EM/liter of water; T4 = 12 ml of EM/liter of water.
Table 4. Feed conversion ratio of pullets placed on different levels of EM.
s. e = standard-error; T1 = control; T2 = 4 ml of EM/liter of water; T3 = 8 ml of EM/liter of water; T4 = 12 ml of EM/liter of water.
Table 5. Mean weekly feed consumption of layers placed on different levels of EM (g/head).
s. e = standard-error; T1 = control, T2 = 4 ml of EM/liter of water, T3 = 8 ml of EM/liter of water; T4 = 12 ml of EM/liter of water.
Table 6. Mean Weekly egg production of the layers placed on different levels of EM.
CV = Coefficient of Variation; Means in a row having similar superscript are statistically not significant (p>0.05); T1 = control, T2 = 4 ml of EM/liter of water, T3 = 8 ml of EM/liter of water; T4 = 12 ml of EM/liter of water.
probiotic (ProtexinTM) containing either 0, 250, 500 or 750 ppm on egg production performance. The researchers reported the highest daily feed consumption from the control group.
Egg production
Age at the first egg of all the treatment groups ranged between 179 and 186 days and there was no significant difference (P>0.05) between all the treatment groups in sexual maturity as measured by the age at the first egg. All the treatment groups seem to be slightly late in sexual maturity, probably attributed to higher body weight attained during the growing (pullet) period. The results obtained also showed that the mean weekly egg production performance of all the treatment groups was low by any standard (Table 6). The mean weekly egg production to an age of 37 weeks of the groups placed on the treatment containing 4 ml of EM/liter of drinking water was significantly higher than all the others (P<0.05). These groups attained daily egg production of 59% (0.59 egg/day/head) at an age of 37 weeks, the value of which was significantly higher (P<0.01) than all the others, indicating that the daily egg production performance of the experimental chicken improved by 12% as a result of administration of 4 ml of EM/liter of drinking water as compared to the control groups. On the contrary, the administration of 8 to 12 ml of EM/liter of drinking water tended to depress mean weekly egg production.
Feed conversion ratio
The amount of feed consumed/ kg or dozen of eggs
produced was lowest (Table 7) for the groups assigned to the treatment containing 4 ml of EM/liter of drinking water indicating that these groups were produced at cheaper rate than all the others (P<0.05). This is further confirmed by the results of the partial budget analysis of laying performance of the experimental layers (Table 9). At present, EM is already commercialized and readily available and in Jimma, a liter of EM is sold at 20 ETB. Assuming daily water consumption of a laying hen at about 250 ml, a liter of drinking water containing 4 ml of EM could economically (0.08 Birr/hen/day) and safely be offered for 4 laying hen/day and it is worth about 0.08 ETB. Market egg price in Jimma is about 2 ETB and the mean daily increment of 0.28 eggs brought with the administration of 4 ml of EM/liter of water is worth about Birr 0.56/hen/day. This shows that the use of 4 ml of EM /liter of drinking water seems to have significant economic implication when used at relatively large scale poultry production.
Egg quality, fertility and hatchability
The results of the egg quality parameters of the eggs collected from the experimental layers are shown in Table 8. There was no significant (p>0.05) difference between all the treatment groups in all the quality parameters considered except in Hough unit and yolk and albumen height, all the three of which were found to be significantly lower (p<0.05) for the groups placed on the control treatment compared to all the others.
The results of this study showed that there was significant improvement in egg quality (Hough unit, yolk and albumen height) with the administration of 4 to 12
Table 7. Feed conversion ratio (feed consumed/ kg or dozen of eggs produced) of the layers placed on different levels of EM.
CV = Coefficient of Variation; Means in a row having similar superscript are statistically not significant (p>0.05); T1 = control; T2 = 4 ml of EM/liter of water; T3 = 8 ml of EM/liter of water; T4 = 12 ml of EM/liter of water.
Table 8. Quality, fertility and hatchability of eggs collected from the layers placed on EM.
CV = Coefficient of variation; Means in a row having similar superscript are statistically not significant (p>0.05); T1 = control, T2 = 4 ml of EM/liter of water, T3 = 8 ml of EM/liter of water; T4 = 12 ml of EM/liter of water.
ml of EM/liter of drinking water. Unfortunately, however, the percentage hatchability reported from this study ranged between 34 and 41% all of which are very low by any standard. There was no significant difference (P>0.05) between all the treatment groups in hatchability. Hatchability and rate of chick survival are one of the major determinant factors of productivity in poultry.
Feed consumption of layers
Significant (P<0.05) difference between the groups of treatment in pullets was recorded after the 1st 5 weeks of the feeding trial. The results of this study are in agreement with that of Kalavathy et al. (2003) who reported improved body weight gain of broiler with supplementary
Table 9. Partial budget analysis on different level of EM (Birr, ETB).
Total cost = cost of birds feed; EM, labor water and electric; Total income = sale of birds and eggs.
administration of Lactobacillus. Mean weekly feed consumption of T1, T2, T3 and T4 833.89, 793.85, 766.07 and 754.36 g head was attained by the groups placed on 0, 4, 8 and 12 ml of EM/liter of drinking water respectively. Similar trend was also reported by Balevi et al. (2009) from the trial conducted to study the effect of dietary supplementation of commercial probiotic (ProtexinTM) containing either 0, 250, 500 or 750 ppm on egg production performance. The researchers reported the highest daily feed consumption from the control group.
Egg production
Weekly egg production on the 13th week of laying was 3.63, 4.13, 3.34 and 3.05 for the treatment groups assigned to 0.4.8 and 12 ml of EM/liter of drinking water respectively. The group receiving 4 ml of EM/liter of drinking water was significantly higher (p<0.01) in egg production than the others. In line with the results of this study, Panda et al. (2008) reported significant increase in the egg production performance of White leghorn layers with dietary supplementation of a probiotic (L. sporogenes) at the rate of 100 mg/ kg−1 diet (6 × 108 spores). However, no further benefit in egg production was noticed by increasing the level of probiotic supplementation from 100 to 150 mgkg−1. Panda et al. (2003) and Kurtoglu et al. (2004) reported that the addition of EM at a rate of 100 or 200 mg/kg of feed resulted in significant improvement in egg production. According to Nahashon et al. (1994) layers fed diets supplemented 0, 1100, and 2200 ppm Lactobacillus produced 88.9, 90.4, and 89.5%, hen-day egg production respectively and the egg production value attained by the groups fed on diet supplemented by 1100 ppm Lactobacillus was significantly higher than that of the control (P<0.05).
Feed conversion ratio
The result showed that treatment level containing 4 ml of EM/liter of drinking water consumed significantly less amount of feed (kg) / kg or /dozen of eggs produced and produced at cheaper rate than all the others (P<0.05).
This is further confirmed by the results of the partial budget analysis of laying performance of the experimental layers (Table 9).
Market egg price in Jimma is about 2 ETB and the mean daily increment of 0.28 eggs brought with the administration of 4 ml of EM/liter of water is worth about 0.56 ETB. This shows that the use of 4 ml of EM /liter of drinking water seems to have significant economic implication when used at relatively large scale poultry production. This result seems to be in line with that of Dahal (1999) who reported that the use of EM (either in water or feed) in broiler production was found to be safe and profitable. Higher profit per bird from the use of EM in water as compared to the use of EM in feed due to additional cost of bokashi preparation was reported by Dahal (1999).
Egg quality, fertility and hatchability
The Hough unit, yolk and albumen height recorded from eggs collected from the groups placed on the control treatment were significantly lower (p<0.05) than that recorded from the eggs collected from all the others. In agreement with this result, an increase in the Hough Units (P<0.05) have been recorded by Daniele et al. (2008) with the use of probiotics. Similarly, Yousefi and Karkoodi (2007), reported improvement in egg quality, as a result of addition of 100 to 750 mg of EM /kg of feed. As shown in Table 8, there were no significant difference between eggs collected from all the treatment groups in fertility and hatchability. The percent fertility of eggs collected from all the treatment groups ranged between 92 and 94%, the values of which are very high by the Ethiopian standard as reported (CACC, 2003; Alemu, 1997 cited in Solomon, 2008). Percent fertility of 75, 80, and 90 was reported from the traditional breeding centers and commercial poultry farms in Ethiopia respectively. On the other hand, there was no significant difference (P>0.05) between all the treatment groups in hatchability.
Hatchability and rate of chick survival are one of the major determinant factors of productivity in poultry. The results of this study agrees with that of Meseret et al. (2011), who reported that the mean percent hatchability calculated for the indigenous chickens of the Gomma Wereda (Jimma Zone) was 22%, the value of which is lower than those reported from different parts of Ethiopia, with the exception of that of Jimma (Tadelle and Ogle, 1996; Mekonnen, 2007). In a trail in which eggs were randomly purchased from Gamma Wereda market places and incubated at JUCAVM along with freshly collected eggs, there was no significant deference between the fresh (27.39) and market (17.63) eggs in percent hatchability. Percent hatchability recorded from both market and freshly collected eggs in Gomma Wereda were very low (Meseret et al., 2011). In summary, the results of this study showed EM could safely and economically be included at 4 ml /liter of drinking water in layers production.
Conclusion
Even though, better egg quality and lower feed consumption were obtained from 8 ml EM/liter of water treated groups due to higher egg production, FCR/dozen of egg, FCR/kg of egg mass and highest profit, 4 ml treatment could provide better production and economic value than any of the treatment levels. Since EM showed insignificant difference for pullet, it is economical not to provide EM for this age group. However, 4 ml of EM/liter of water showed better performance of egg production and egg quality, provision of this amount of EM fifteen days before onset of egg lay up to the end of production period would be economical. Since weight gain of females RIR growers performed better at 8 ml of EM/liter of water while males RIR grew best at 12 ml of EM/liter water. There is a need for further investigation to determine such level of EM for broiler type breeds.
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The Effects of Supplementation of Effective Microorganisms on Egg Production Traits, Quality Parameters and Chemical Analysis During the Late Laying Period in Hens
Authors
El-Deep, M. H.; Amber, K.; Sayed, M. A. M.
Editors
Köfer, J.;Schobesberger, H.
Conference paper
Animal hygiene and sustainable livestock production. Proceedings of the XVth International Congress of the International Society for Animal Hygiene, Vienna, Austria, 3-7 July 2011, Volume 3 2011 pp. 1267-1269
Conference Title
Animal hygiene and sustainable livestock production. Proceedings of the XVth International Congress of the International Society for Animal Hygiene, Vienna, Austria, 3-7 July 2011, Volume 3.
ISBN 978-80-263-0012-0
Record Number: 20113298498
ABSTRACT
The effects of effective microorganisms (EM) on the Egg production traits, Egg quality and egg chemical analysis of layers were studied. Five hundred and forty Inshas layers (a local Egyptian chicken strain) were randomly divided into 6 groups with 90 layers in each group. Layers in group 1 were fed a control diet. The remaining groups received the control diet that contained 2.5, 5.0, 7.5 or 10.0 ml of EM/kg, and 20 mg of zinc bacitracin/kg respectively. The obtained results clarified improvements in egg production (Egg number and Egg mass/hen) and Egg quality ( P<0.01) of layers when EM was added to the diets. The results also showed that, egg yolk cholesterol was significantly decreased in chicks that fed diets with different levels of EM as compared with the control diet, while egg protein percentage was significantly (p≤0.05) increased in birds fed diets with different levels of EM.
Using an Effective Microorganism Supplementation in Layers
Sasitorn Chotisasitorn, Somchai Chantsavang, Seksom Attamangkune, Attawoot Plaiboon
KASETSART JOURNAL: NATURAL SCIENCE, Volume 031, Issue 3, Jul 97 - Sep 97, Page 363 - 367
For full article, visit: http://research.rdi.ku.ac.th/world/kjournal.php?journalid=4&articleid=1741&webLang=en
Keywords: layer, effective microorganism, calcium, specific gravity.
Abstract
An experiment was conducted to study the effects of supplementation of EM ( effective microorganism ) in feed on laying performance and egg quality. A 2x3 Factorial in Completely Randomized Design with 4 replications, using the total numbers of 288 layers was utilized. In one factor, supplementation and non-supplementation of EM were applied. In the other factor, supplementation of calcium at levels of 3, 3.5 and 4% were used. Results of the study, over the 3 period, 28 days per period revealed that there were no significant effects of supplementation of EM on daily feed intake, body weight gain, mortality, egg mass, egg weight, albumin weight, yolk weight, egg shell weight, yolk color and haugh unit (P>0.05). But egg production (P<0.05), feed per 1 dozen egg and specific gravity were highly significant difference (P<0.01).
Key words: layer, effective microorganism, calcium, specific gravity
INTRODUCTION
Several reports in the literature have been found pertaining to the influence of microorganism on animals for high efficiency in animal production. Probiotic is used for feed supplementation. Males and Johnson (1990) defined the term "probiotic" to be used for life encouragement by microorganisms. They are living cells and by-products from microbial fermentation. The types of probiotics range from pure culture such as Lactobacillus acidophillus or mixture fermentation such as a mixed culture of organisms of fungi and bacteria or fungi and yeasts or other combinations in which the components are quite unrelated. (Hesseltine, 1991)
EM ( effective microorganism) is the mixed cell culture composed of photosynthetic bacteria, actinomyces, yeast, Lactobacillus and fungi (Higa, 1993) Chantsavang et al. (1993) reported the effects of EM supplemented in drinking water and feed of Japanese quails that there was no significant effect of EM on growth, feed efficiency and mortality rate of growing quails and laying period (4-12 wks) but there was significant effect in egg quality trait and high content of crude protein in manure. While research with Lactobacillus cultures has focused primarily on disease processes, Tortuero (1973) found that a Lactobacillus probiotic and zinc bacitracin had similar effects in stimulating weight gain and feed efficiency in broiler chicks. Chapman (1988) reported that the probiotic caused a distinct change in microbial flora of the ceaca and small intestine in that by nine days of age, enterococci had essentially disappeared. According to the similar results of enhanced gain and (or) efficiency of broilers in response to probiotic culture was reported by Crawford (1979) and Dilworth and Day (1978).
Yeast culture is considered to use for enhanced feed utilization. Martin (1995) reported that using of Lacto-Sacc (Alltech) 1 kg/t in broiler decreased 6.4% female mortality and 23.5% male mortality. Egg production (per hen day) in the period was improved from 59.9% to 63%. Layer should get optimum calcium at level 3.6 g/hen/day or 3.5% in feed. This experiment was conducted to determine the effects of Effective Microorganisms supplementation in layer feed which would have any beneficial effects on productive performance.
MATERIALS AND METHODS
The experiment was conducted in during 3 periods, each lasting 28 days with a total of 288 Isa Brown Layers, approximately 70 weeks of age. A 2 x 3 factorial in Completely Randomized Design with 4 replications (twelve individually cages hens per replicate) was used.
In one factor, supplementation and non-supplementation of 1% EM were applied. In the other factor, supplementation of calcium at the levels of 3, 3.5, and 4% were used. All birds were fed with the diets (Table 1) and water ad libitum throughout the entire study. Daily feed intake, feed per 1 dozen egg, mortality, body weight gain, and egg production were determined. Egg mass, specific gravity, albumen weight, yolk weight, yolk color, haugh unit, egg shell weight, and egg shell thickness were collected from a 3-day egg sample taken from each replication at the end of each 28-day period. Natural light was supplemented with artificial light in the morning and afternoon to provide a 16-hr photopenod. The treatments were as follows:
Treatment 1: 3% of calcium level in feed and 1% of EM supplementation
Treatment 2: 2% of calcium level in feed
Treatment 3: 3.5% of calcium level in feed and 1% of EM supplementation
Treatment 4: 3.5% of calcium level in feed
Treatment 5: 4% of calcium level in feed and 1% of EM supplementation
Treatment 6: 4% of calcium level in feed
Data was subjected to an analysis of variance through the Statistic Analysis System procedures (SAS, 1988).
TABLE 1 Composition of diets for layers
RESULTS AND DISCUSSION
The data is presented in Table 2 and 3. Results showed that there were no significant difference on daily feed intake, body weight gain and mortality (P>0.05) in supplementation and non-supplementation of EM in any level of calcium.
Similar report of Miles et al. (1981) found that there were no significant differences in feed consumption, egg production and mortality in adding a Lactobacillus culture to the diet of Bobwhite quail breeds. Damron et al. (1981) also reported that 625 mg/kg of Probios (a mixed Lactobacillus culture) in turkey hen's feed did not significantly affect egg production, daily feed intake, specific gravity of eggs, or body weight. Miles et al. (1981) remarked that the addition of the Lactobacillus culture at a level of 0.625 g/kg feed in bobwhite quail breeders had no significant influence on feed consumption, egg production, fertility, hatchability of fertile eggs and mortality.
However, it was found that there was a highly significant difference (P <0.01) in feed per dozen egg by 3% calcium level consumed feed more than 3.5% and 4% calcium level respectively and there was interaction between 1st factor and 2nd factor in feed. 3.5% calcium level without EM supplementation (T4) had significant difference (P<0.05) in egg production (%) therefore EM did not affect calcium utilization in feed for production.
Table 2 Effect of EM supplementation in feed at different calcium levels on layers
abc: Mean values on the same row with common superscripts are highly different at (P<0.01)
xyz: Mean values on the same row with common superscripts are different at (P<0.05)
Table 3 Effect of EM supplementation in feed at different calcium levels on egg quality
abcd: Mean values on the same row with common superscripts are highly statistically different (P<0.01)
Table 3 shows no significant effects (P>0.05) on egg mass, egg weight, albumen weight, yolk weight, egg shell weight, yolk color, haugh unit in every treatment, which was similar to the report of Boonyoung et al. (1995) who also found that there were no difference in egg mass, egg weight, albumen weight, yolk weight, egg shell weight, egg shell thickness, yolk color, haugh unit and specific gravity in EM supplement in drinking water of layers.
In the variation of calcium levels at 3%, 3.5% and 4% Table 3 shows specific gravity values vary to calcium levels. 3.5% calcium level had the highest specific gravity value (P<0.01). Although in this experiment, layers were raised during summer season, egg production did not decrease. It may be because of evaporation housing system. The temperature in house was about 27oC and good condition for raising layers. The layers fed with 3% calcium produced egg (P<0.05) with thinner egg shell weight (P>0.05)
CONCLUSION
1. There were no significant differences on daily feed intake, weight gain, mortality, egg mass, egg weight and yolk weight in layers at 3%, 3.5% and 4% calcium levels.
2. There were highly significant differences on feed per dozen egg and specific gravity (P<0.01), egg production (P<0.05).
ACKNOWLEDGMENTS
The authors are grateful to Poultry Research and Development Center, Kasetart University Research and Development Institute for financial support, Agriculture Center of Kusae Saraburi for EM support.
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