INTRODUCTION

Nature and Importance of the Study

Probiotics is a term which means “for life” and defined as live microorganisms that beneficially affect the host by improving the balance of the intestinal micro flora”. It is commonly used to refer to “good bacteria” that one has to have in the body in order to maintain a healthy immune system. Mc Cann et al., (2004)

There are over 400 different known species of bacteria that inhibit the normal bowel-excluding viruses, yeast and parasites. To put it in another way, there are 20 times more bacteria in the body than there are in cells. And more than the total number of people who in the gut is very important since the intestinal tract is home to some 100 trillion microorganisms, but only a few of them are friendly. Mc Cann et al., (2004)

Research suggests these benefits are due in part to the effect s probiotic bacteria have on a wide range of intestinal functions. Several types of probiotic culture (Lactobacillus casei, Lactobacillus acidophilus, Lactobacillus plantarum, Bifidobacterium) are being introduced in an expanding variety of dairy product round the world. Daily consumption of probiotic foods, such as yogurt and cultured dairy drinks, may be beneficial in one of several ways. Gill et al (2004).

Some of these probiotics help strengthen the body’s defenses by providing a regular source of probiotic bacteria for the intestinal tract; some might provide a much needed calcium for individual who are unable to consume most dairy foods because of lactose intolerance; and some may help to correct a potential balance of the intestinal microflora Adofsson, et.al, (2004). Several studies have shown that regular consumption of some probiotics helps maintain the balance of the intestinal microflora which can have a wide range of beneficial effects.

The use of growth- promoting antibiotics is being placed under more and more pressure as consumers increasingly fear that their use in feed ration of productive livestock lead to the formation of resistance against bacteria which are pathogenic to humans. Simultaneously, the adverse effect of antibiotic feeding has encouraged a shift In favor of feeding probiotics to be used exclusively as growth stimulants and for improvement of the feed conversion rate to boost up productive performance of chicken Langhout, (2000).

Poultry nowadays are are raised for intensive production system in densely populated flocks. During this process, chicken may get stress from a number of factors such as overcrowding, unfavorable ambient, feed intake and vaccination. The dietary use of probiotics feeding is gaining momentum in broiler to counteract and to minimize the stresses Cavazonni, et. al, (1998)

The biggest challenges still arise in determining the correct type and quantity of probiotics because of the number and diversity of microbes and the poorly understood interaction between the microbes and intestines. Hence, this study will be conducted to find out the influence of the different kinds of probiotics, L. casei, L. acidophilus and L. plantarum as feed supplement for broilers.

The result of this study will help the researcher and other neighboring communities to have adequate knowledge and genuine information on the uses of these probiotics as feed supplement

Objectives of the Study

Generally, this study aims to find out the effects of the three (3) kinds of probiotics, Lactobacillus casei, Lactobacillus acidophilus and Lactobacillus plantarum on the growth performance of broilers.

The specific objectives of the study are as follows:

1. To determine the effect of different probiotics on the   growth performance of broiler in terms of weight gain, feed conversion efficiency, amount of feed consumed and volume of water consumed.

2. To determine the incidence of diseases.

3. To determine the cost and return in broiler production with the use of probiotics

Hypothesis of the Study

There is no significant effect on the growth performance of broiler using the three (3) different kinds of probiotics in the drinking water in terms of feed consumption, feed efficiency, and feed consume ratio, weight gain and water intake.

Scope and Limitations of the Study

The study will focus on the growth performance of broiler supplemented with three (3) different probiotics, Lactobacillus casei, Lactobacillus acidophilus and Lactobacillus plantarum. Weight gain of chicks, feed consumption, water intake, feed conversion efficiency and mortality rate will be recorded. Also, net income will be determined and analyzed. The data on carcass yield, meat cut and intestinal pH will not be gathered. Drinking water will be supplemented with probiotics preparation in the form of solution.

A total of 96 day old broiler chicks will be reared under hygienic management practice throughout the entire of the study.

Review of Related Literature

To support this research work and to provide readers with essential information, related literature and studies about the incorporation of probiotics in drinking water for chicks were reviewed to determine its effectiveness to the study.

Beneficial Effect of Probiotics

      The major consumption of probiotics is in the form if tablet or powder but most commonly in dairy based food containing intestinal species of Lactobacilli and Bifidobacterium. Probiotics will be referred to as supplement of animal feeds for farm animals but it is easily applied to humans. The are many companies and scientists that showed interest in establishing scientific credibility in probiotics

      A Nobel- prize winner, Ellie Metchnikoff (1845-1916) is considered to have been the inventor of probiotics. He was the first one who suggested that consuming bacteria could have a beneficial effect on health. He proposed that the acid producing organisms in fermented dairy product could prevent what he called “fouling’’ in the large intestine and , if consumed regularly it will  lead to longer , healthier life.  Bonci (2004)

Probiotics in Animal

      Modern rearing method which includes unnatural rearing condition and diet induce stress and can cause changes in the composition of the microflora which compromise the animals’ resistance to infection. The aim of the probiotics approach is to repair the deficiencies in the microflora and restore the animals’ resistance to disease. Such treatment does not introduce any foreign chemicals into the animal’s internal environment and does not introduce any foreign chemical into the animal’s internal environment and does not run the risk of contaminating the carcass and introducing hazardous chemical into the food chain.

      Probiotics are now replacing the chemical growth promoter for farm animals and claim have also been made for increasing resistance to disease. The benefits claimed for probiotics in farm animals increased growth rate, improved feed conversion, improved resistance to disease improved milk yield and quality and improved egg production, Fuller (2004)

Lactobacillus acidophilus

      Lactobacillus acidophilus may be considered a probiotic or “friendly” bacteria. This type of healthy bacteria inhibits the intestine and vagina which protect against some unhealthy organisms. The breakdown of nutrient by Lactobacillus acidophilus produce lactic acid, hydrogen peroxide and other byproduct that make the environment hostile for undesired organism Lactobacillus acidophilus also tends to consume the nutrients other microorganism depend on, thus outcompeting possibly harmful bacteria in digestive tract. During digestion, Lactobacillus acidophilus also assists in the production of macin, folic acid and pyridoxine L. acidophilus can also assist in bile deconjugation, separating amino acid from bile, which can then be recycled by the body.

      Some research have indicated Lactobacillus acidophilus that it may provide additional health benefits including improved gastrointestinal function, boosted immune system and decreased in the frequency of vaginal yeast infection. Some people reported that Lactobacillus acidophilus provides relief from indigestion and diarrhea. University of Nebraska study found that feed supplement with Lactobacillus acidophilus and fed to cattle resulted in 61% reduction of Escherichia coli. Research has indicated Lactobacillus acidophilus helpful in producing serum cholesterol level (http://eu.wikipedia.org/Lactobacillus _acidophilus).

      In vivo administration of Lactobacillus acidophilus improved growth performance of weaning pig was observed. Additionally Lactobacillus acidophilus works as probiotic which increases the number of desirable microflora in the gut by decreasing intestinal pH. Reports stated that Lactobacillus acidophilus increased ADG and feed efficiency of pig fed diet supplemented by lactose. http://www.ansi.okstete.edu/reseach/2001rr/42/.42.htm

Lactobacillus casei

      Lactobacillus casei is a transient, anaerobic microorganism of genus Lactobacillus found in the human intestine and mouth. As a lactic acid producer, it assisted in the propagation of desirable bacteria. This particular species of Lactobacillus is documented to have a wide pH and temperature range and complement the growth of Lactobacillus acidophilus, a producer of the enzyme amylase. It is known to improve digestion and to reduce milk intolerance.

      The most common application of Lactobacillus casei is industrial specifically for dairy production; however, a team of scientist from Simon Bolivar University, Caracas, Venezuela found out that Lactobacillus casei bacterium in bean natural fermentation, the bean contains lower amount of the compounds causing flatulence upon digestion (http://eu.wikipedia.org//Lactobacillus_casei)

      According to Consinagdo (1992), stated in his study that he did an experimental to determine the antibacterial activity of common diarrhea using ETEC, Lactobacillus casei against Salmonella enteritidis, Shigella dysenteriae, Vibrio cholerae and to determine the effective contact that Lactobacillus casei exerts bacterial activity against these organisms.

      Thus, clinical study using Lactobacillus casei as adjunctive or primary management of diarrhea due to mentioned pathogen is highly recommended.

Lactobacillus plantarum

Lactobacillus plantarum has one of the largest genomes known among the lactic acid bacteria and is very flexible versatile species. Lactobacillus plantarum which is related to lactbacilli is unusual in that it can respire oxygen but has no respiratory chain. The consumed oxygen ultimately ends up as hydrogen peroxide. The peroxide probably acts a weapon to exclude completing bacteria from the food source. In place of the protective enzyme superoxide dismutase is present in almost oxygen-tolerant cell. The organism accumulates millimolar quantities of manganese polyphosphate. Because of chemistry which manganese compreses protects the cell from oxygen damage is subverted by iron. These cells contain virtually no iron atom; In contrast a cell of Escherichia coli of comparable volume contain over one million iron atom.

Lactobacillus plantarum is the most common bacterium used in silage inoculants.

The study of Mogol (2006) revealed that the use of 20ml/1 liter of probiotics Lactobacillus plantarum as supplement gave the highest weight gain in birds.

Methods

Experimental Design

A Randomized Complete Block Design (RCBD) will be used in this study. The weight of the chicks will served as the determinant factor in assigning into them to different blocks. There will be four treatment assigned to be assigned in each block.

The treatments to be added to chick’s drinking water are follows

T1= 1000ml water

T2= 1000ml water+ 20 ml Lactobacillus acidophilus

T3= 1000ml water+ 20 ml Lactobacillus casei

T4= 1000ml water+ 20 ml Lactobacillus plantarum

Preparation of Equipment and Facilities

All the needed equipment and facilities that will be used in the study will be prepared a week before the arrival of the chicks. The poultry house will be cleaned off from dust and foreign matters.  The experimental cages, brooder, feeding troughs, waterer and other equipment will be cleaned using washing with soap and water. After washing, the material will be disinfected using disinfectant.

Procurement of Stock

The ninety- six (96) heads of one-day-old chicks will be purchased from reputable distributor in Pinamalayan, Oriental Mindoro. The chicks will be placed in a clean box with holes and will be provided with enough space to ensure their comfort are to avoid from injuries due to overcrowding that may occur during travel.

Brooding

      Upon arrival the chicks will be brooded immediately for two weeks. At this period, brooder will be provided with four 50 watt incandescent bulbs to keep their bodies warm and comfortable. Clean old newspaper will be laid in the floor to conserve heat and to serve as surface where feeds will be placed. When temperature gets high, the birds or chicks will open their beak and will spread their wings. Eating habit will be lessened and will remain inactive. In this condition the temperature will be adjusted downward. When chicks crowded together, the source of heat will be lowered to decrease the temperature. During brownouts, kerosene lamps will be used as source of light and heat.

Feeding Management during Brooding

During brooding stage, the chicks will be provided with Water and Commercial feeds in ad libitum way. The chicks will be fed with chick booster with drinking water for the first two weeks and will be provided with electrolytes, this will be done at 6:00 A.M. daily.

Distribution of chick to respective cages

Initial weight of chicks will be gathered right after the brooding period. The weight of chicks will be ranked in descending order. The first 24 heaviest chicks will be randomly assigned at to the four cages in Block I. Rank 25-48 will be assigned to the Block II. Same procedure will be done in assigning birds ranked 49-72 and 73-96 for Block III and IV respectively.

Feeding of experimental birds

Feeding of broiler starter crumble will start during the growing period until two weeks. After 30 days of brooding broiler finisher will be given up to the end of the study.

Supplying of Water to the Experimental Birds

Birds in treatment will be given 1000 ml of pure water and the rest will be incorporated with 20 ml Lactobacillus acidophilus, 20 ml, Lactobacillus casei, 20 ml, Lactobacillus plantarum and to given ad libitum. This will be done until the end of the experiment.

Lightning Management

During the third week (growing stage) the chicks will be provided with light from electrical source from 6p.m.-6a.m. using 50 watt incandescent bulb to stimulate feeding at night, protect them from predator and maximize weight gain. Kerosene lamps will be provided during brownout.

Gathering of Data

Body Weight

The initial weight of birds will be determined at the end of the brooding period. Subsequent weighing will be done weekly in the morning before feeding the bird. The body weight gain of the experimental birds will be obtained by subtracting the first week’s weight. This system of the body weight gain will be continued up to the end of the feeding experiment.

Feed Consumption

All leftover feeds will be weighed and the weight obtained will be subtracted from the daily feed given to the birds. The difference will be recorded as daily feed intake.

Amount of Water Consumed

With the use of graduated cylinder, all unconsumed water will be measured and subtracted from the water given daily to the birds. The difference will be noted as daily water intake and will be recorded weekly. Recording of water intake will start after breeding.

Feed Conversion Ratio

The feed conversion ratio per treatment will be obtained by dividing the average feeds consumed by the average weight gain of birds times one hundred which will be done weekly until the end of the experiment.

Feed Conversion Efficiency

      The feed efficiency per treatment will be obtained by dividing the total weight of feeds consumed by the total weight gain of feeds

Morbidity and Mortality

      In getting the mortality rate of broiler, the number of dead birds will be recorded. Likewise, the number of those showing illness will be recorded throughout the duration of the study.

      The percentage of morbidity per treatment will be computed by dividing the number of broilers that acquired disease during the experiment by the number of broilers per treatment and multiplying the value by 100. Percentages of mortality per treatment will be computed by dividing the number of dead broilers by the number of broiler per treatment and multiplied by 100.

Analysis of Data

The data gathered will be analyzed using analysis of Variance (ANOVA) for the Randomized Complete Block Design (RCBD) and will be tested at 1% and 5% levels of significance. Duncan’s Multiple Range Test (DMRT) will be used to determine the significance among treatment means.

RESULT AND DISCUSSION

Average Initial Weight                                                                    A total of 96 day-old chicks were used. Before the chicks were transferred to their respective experimental cages after the brooding period, they were grouped according to their initial weight which served as the blocking factor. Weight gain of birds was the basis of determining the growth performance.       Table 1. shows the averages initial weight of birds. There was a big difference in  variation observed in the body weight of birds among the treatments Birds supplied with 1000 H2O+ 20ml acidophilus(T2) in the drinking water gained an average 0f  344.17 compared to birds given only 1000 H2O+ 20ml casei  (T3) in their drinking water with a numerical difference of 67.916g. However, when data were subjected to ANOVA test no significant difference variation was observed (Table1.1) The significant difference among block was result process of classifying birds into blocks in accordance with their weight. The highest weight was assigned in Block I and the lowest weight was assigned in block IV.

Table. 1 Average initial weight of broilers (g)

Table 1.1 Average initial weights of broilers  

Average Body Weight Gain of Broiler after One week of Feeding                                   After a week of observation, increase in weight gain of birds was evident (Table 2). Birds supplied with H2O+ 20ml plantarum (T4) in their drinking water gained an average of 459.17 compared to the birds given with H2O+ 20ml acidophilus (T2) with the numerical difference of 67.92g. However, when data were subjected to ANOVA test no significant variation was observed (Table 2.1)                                      The result appeared that the different kinds of probiotics did not give any remarkable increase in the weight of the bird because the result was still comparable with that of the control. This might be attributed to the fact that the birds were making adjustments from their transfer from the brooding cages to the experimental cages and changes in the feed given from booster to starter

Table. 2 Average weight gain of broiler after one week of feeding (g)

Table 2.1 ANOVA on average weight gain of broiler after one week of feeding  

Average Body Weight Gain of Broiler after two weeks of Feeding                                  After two weeks of feeding, the change in ranking in weight gain was observed (Table 3). Birds supplied with plain H2O+ 20ml acidophilus (T2) in their drinking water gained an average of 556.25g compared to birds given with plain water only (T1) the numerical difference of 47.92                             Statistical analysis showed that the difference among treatment were insignificant (Table 3.1).  Findings showed that different kinds of probiotics added to the drinking water of birds were still comparable with a control. The different kinds of probiotics did not exert any advantages I terms of weigt gain after the second week of the experiment.

Table 3. Average weight gain of broiler after two weeks of feeding (g)

Table 3.1 ANOVA on average weight gain of broiler after two week of feeding  

Average Body Weight Gain of Broiler after three weeks of Feeding                                After third week of feeding (Table 4) changed in ranking of treatment in terms of body weight gain of broiler was observed. Birds supplied plain water only (T1) in their drinking water gained an average of 670.83 compared to birds given with H2O+ 20ml casei (T3) with the numerical difference of 60.42g.           Insignificant differences among treatments were noted. The probiotics supplied to the body of the birds again did not exert any advantages in terms of weight gain after the third week of the experiment. Results showed that the effect of different kinds of probiotics incorporated to the drinking water were comparable with the control (T1).

Table 4. Average weight gain of broiler after three weeks of feeding (g)

Table 4.1 ANOVA on average weight gain of broiler after three weeks of feeding     

Average Body Weight Gain of Broilers after Four Weeks of Feeding                                      On the last week of incorporating probiotics in the diet, birds supplied with H2O+ 20ml casei  (T3) gained an the average of 539.58g compared to birds given H2O+ 20ml acidophilus (T2) in the drinking  water with numerical difference of 93.75 (Table 5). However, analysis of variance indicated no significant difference in all treatment (Table 5.1) 

Table 5. Average weight gain of broiler after four weeks of feeding (g)

Table 5.1 ANOVA on average weight gain of broiler after four weeks of feeding

Average Final Weight of Broilers after four Weeks of Feeding                                          Data for the mean final weight of broiler (Table 6.) showed that birds supplied with H2O+ 20ml plantarum (T4) in the drinking water gained an average of 2462.50g compared to birds that were H2O+ 20ml casei (T3), with a numerical difference of 64.58g.                                            ANOVA for final weight of broilers after four weeks of feeding showed no significant difference among treatment (Table 6.1)

Table 6. Average final weight of broiler (g)

Table 6.1 ANOVA on average final weight of broiler

Average Weekly Weight Gain of Broilers                                                    Table 7. shows that weekly gain of broilers from the first to fourth week of feeding . At the end of the study, birds in H2O+ 20ml plantarum (T4) had the highest weight gain with 527.81g compared to the H2O+ 20ml acidophilus with a numerical difference of 48.66g (Table 7). It was noted that probiotics may present advantage in increasing resistance to disease, increasing growth rate and improved feed conversion, as Fuller(2004) conformed. However analysis of variance shows that there was no significant difference among treatment.

This result means that the probiotics added to the drinking water of birds were comparable with control (T1).

Table 7. Average final weight of broiler (g)

Table 7.1 ANOVA on average final weight of broiler

Figure 1. showed that during the second week of birds, a slight increase in the body weight was observed at T1, T2, T3 and T4. Meanwhile, during the thirds week of weighing birds T1, T2, T3, and T4 indicated highly increase. However, during the fourth week of weighing birds, T3 and T4 indicated a slight decrease on body weight while T1 and T2 showed a highly decrease on the body weight.

Figure 1. Average weight gain of broiler

Feed Consumption during the First Week of Feeding                                               On the first week of observation, it was observed that birds H2O+ 20ml casei (T3) had the highest mean with 3470g, compared to birds given with H2O+ 20ml acidophilus (T2) in their drinking water with the numerical difference of 135g. When the data were subjected using Analysis of Variance Test, insignificant differences among treatments had been observed (Table 8.1)                                           The result appeared that the different kinds of probiotics did not give any remarkable increase in the weight of the bird because the result was still comparable with that of the control (T1). This might be attributed to the fact that the birds were making adjustments from their transfer from the brooding cages to the experimental cages and changes in the feed given from booster to starter crumble.

Table 8. Feed consumption during the first week (g)

 Table 8.1 ANOVA on feed consumption during the first week

Feed Consumption after during the Second Week of Feeding                                        At this point of the experiment, a sudden change appeared in the ranking of the birds in terms of feed consumption. H2O+ 20ml plantarum (T4) had the highest mean with 5427.5g compared to birds given with H2O+ 20ml acidophilus (T2) with a numerical difference of 100g (Table 9)                                       Table 9.1 shows that the recorded feed consumed was not statically at all.                         This result showed that the probiotics added to the drinking water of birds were comparable to the birds given plain water (T1). The treatment applied did not affect the feed consumed by the birds.

Table 9. Feed consumption during the second week (g)

Table 9.1 ANOVA on feed consumption during the second week

Feed Consumption during the Third Week of Feeding                                               On the third week of incorporating probiotics in the diet, it was observed that birds in H2O+ 20ml plantarum (T4) had the highest feed consumed with 5462.5g compared to birds given with H2O+ 20ml acidophilus (T2) with a numerical difference of 1025.g

      Table 10.1 shows that the recorded feed consumed was not statically at all.                           This result showed that during the third week of the experiment, probiotics were comparable with the control (T1). The treatment did not affect the feed consumed by the birds which resulted to minimal difference on increase in feed consumption among treatment.

Table 10. Feed consumption during the third week (g)

 Table 10.1 ANOVA on feed consumption during the third week

Feed Consumption during the Fourth Week of Feeding                                              On the last week of incorporating probiotics to the diet, the change in trend of ranking in feed consumption was noticed. Birds supplied with H2O+ 20ml acidophilus (T2) in the drinking water in the drinking water gained an average of 5175g compared to birds given H2O+ 20ml plantarum (T4) with a numerical difference of 600g. However, analysis of variance for feed consumption of broiler on the fourth week indicated no significant differences among treatment.                                                  This result showed that the probiotics added to the drinking water of birds did not affect the feed consumed of the broilers, the 1000 H20 + 20ml acidophilus (T2), 1000 H20 + 20ml casei (T3) , 1000 H20 + 20ml plantarum (T4) were comparable with the control T1

Table 11. Feed consumption during the fourth week (g)

Table11.1 ANOVA on feed consumption during the fourth week

Total Feed Consumption                                                              Collecting data on weekly feed consumption started after transferring the birds to the experimental cages. The accumulated feed intake in four weeks is presented in Table 12.                           The   total amount of feed consumed by 96 birds during the experiment was 294.090kg. Table 12 shows that the highest amount of feed was on the first week which was obtained by birds given H20 + 20ml plantarum (T4) with 75.500 kg compared to birds supplied with plain water only (T1) with a numerical difference of 3.780 kg. Nevertheless, the difference in feed consumed was affected by varying different kinds of probiotics, as observed. The results indicate that the total amount of feed consumed had something to do with the water consumption. The more the birds consumed water the more they were likely to consume feeds. ANOVA (Table 12.1) for feeds consumption indicates no significant differences in all treatment.

Table 12. Feed consumption from first week to fourth week of the study (g)

Table12.1 ANOVA on total feed consumption from first to fourth week of the study (g)

Figure 2 showed that during the second week of feed consumption of birds, a slight increase on feed consume was observed at T1, T2, T3, and T4. Meanwhile, during the third week of feed consumption of birds, T1, T2, and T3 indicated a slight decrease on feed consumed while T4 exhibit a slight increase on feed consume. However, during the fourth week of feed consumption of birds, T2 indicate a slight increase on feed consumption on feed consumed while T1, T3, and T4 showed a slight decrease on feed consumed.

Feed Conversion Ratio                                                                     The weekly feed conversion of broiler fed with probiotics incorporated to the drinking water of birds from the first to fourth week of feeding experiment is shown in Table 13. The highest feed conversion ratio was obtained by birds fed with water was obtained by birds fed with plain water only (T1) with a mean of 1.45, followed by H20 + 20ml casei (T3) with a mean 0f 1.48, then H20 + 20ml plantarum (T4) with 1.49  compared to birds given with H20 + 20ml acidophilus (T2). It showed that water only (T1) was the most efficient feed converter as compared to the rest of the treatments but when data were analyzed using ANOVA, no signidicant variation was observed.                                                                   Table 13.1 shows that the variation in feed efficiency was not due to the treatments since the computed F-values did not equal nor exceed both stated of confidence.

Table 13. Average feed conversion ratio

Table13.1 ANOVA for the average conversion ratio