Field trial to study the possibilities of using probiotic bacteria in drinking water replacing antibiotics/growth promoters in feed during broiler rearing and to investigate the effect of catalyst activated water (CAW) when combined with the probiotic in drinking water

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Antibiotics are commonly used in food animals to promote growth and prevent disease, as well as to treat sick animals. This has led to the development of antibiotic resistant pathogens and subsequently different countries have banned the use of antibiotics as growth promotion (AGPs). To eliminate the use of AGPs and to maintain animal productivity, some farmers have been using live microorganisms in the feed as feed supplements which beneficially affects the host animal by improving its intestinal microbial balance. Benefits from probiotic microorganisms have been recognized for over 100 years, and as being useful in poultry for about 50 years. The most commonly used probiotic microorganisms include members of lactic acid bacteria (LAB), like Lactobacillus spp., Bifidobacteria spp. and Saccharomyces spp.  which do not produce heat resistant spores and vegetative stages of these species are easily destroyed by the heat applied during the feed manufacturing process.

Bacillus species are gram positive, rod shape bacteria that are strict aerobes or facultative anaerobes which produce heat resistant spores under stressful conditions. Though the soil is considered the habitat, several studies have shown that Bacillus spores can be found in the intestinal tracts of animals where they live and multiply actively. There is enough literature to suggest that the Bacillus spp. is commensal bacteria of the gastrointestinal tract. Though, there are published literature on the performance of broilers with feed supplemented with spores Bacillus spp., there is hardly any on the use of Bacillus probiotic bacteria delivered via broiler drinking water and the present study was to study the possibilities of replacing the use of antibiotic/ growth promotors in the feed and delivering a mixture of Bacillus species through drinking water. Also, tested the effect of catalyst activated water on the broiler performance.    

Objectives

  • To study the effects of using a mixture of five Bacillus species including, 2 strains of Bacillus subtilis, B.licheniformis, B.coagulans, and B.amyloliquefaciens isolated from local environments on the performance of broiler rearing.
  • To study the effect of catalyst activate water (CAW) on the performance of broiler rearing.

 

Experimental design

Day old cull (unhealed naval) broiler chicks obtained from a reputed hatchery operator in Sri Lanka were used for the trial. Chicks were randomly grouped into four groups, each group having 31 chicks. Three groups were used as treatments; T1, T2 and T3, and one group was used as the control (positive control). Conventional open house rearing with bird space of one square foot per bird was employed. The feed used for the treatment groups was free from antibiotics/growth promoters and the drinking water was free from chlorine. The birds in the control group received normal feed which was available for the other broiler farmers containing antibiotic/growth promoters. Drinking water for the control group was treated with chlorine (2 mgL-1). Chicks in the control group were administered through drinking water with an antibiotic during the first 3 days. The probiotic bacterial count of the water for the three treatments was 2.44 x 103 CFU ml-1. The CAW was diluted according to the instructions given on the bottle (200 ml of CAW to 1000 liters of water). The four groups of birds received treatment as given in table1.

Table 1. Treatments of the four group of chicks ( + treated, - not treated)

Birds were weighed weekly, mortality and the feed conversion ratios were calculated at end of the trial, 42 day period. Analysis of variance (ANOVA) was carried out to find out whether the growth of birds of treatments groups was different from those of positive control group.

 

Results

Figure 1 shows the average weekly growth of birds of different groups and the growth differences were not significant from each group.

Fig. 1.  Effect of probiotic bacteria and CAW given through drinking water on the average growth of broiler birds compared to positive control. (T1 – Probiotic, T2 – Probiotic + CAW, T3 - CAW, Positive Control – Feed with antibiotics/growth promoters & chlorinated water). 

Fig. 2.  Effect of probiotic & CAW on FCR compared to positive control; (T1 – Probiotic, T2 – Probiotic + CAW, T3 - CAW, Positive Control – Feed with antibiotics/growth promoters & water chlorinated water). 

The highest FCR was observed (2.13) for the positive control group while the lowest (1.97) was for both T2 & T3 groups. The group that received only probiotic had an FCR of 2 (Fig.2).

Fig. 3. Effect of probiotic &CAW on the mortality of broiler birds compared to positive control; (T1 – Probiotic, T2 – Probiotic + CAW, T3 - CAW, Positive Control – Feed with antibiotics/growth promoters & water chlorinated water).

The highest % mortality (12.91%) was recorded for the positive control group (chlorinated water, no probiotic bacteria in drinking water, feed containing antibiotics/growth promoters) while all the three treatment groups had percentage mortality of 3.23% (Fig. 3).

The two groups of chicks that were treated with the probiotic bacteria had wet litter during the first three days which slowly disappeared after 3 days but the chicks took feed and water normally during the wet litter period. 

 

Conclusion

  • Rearing broiler could be done without antibiotics/growth promoters in feed when a mixture of Bacillus species are given via drinking water.
  • When probiotic bacteria are used, early antibiotic treatments (prophylactic doses vis drinking water) are not necessary.
  • The performance of broilers with Bacillus species in drinking water is better than that is observed under conventional broiler rearing (with antibiotics/growth promoters in feed, chlorinated water and prophylactic antibiotic treatments).
  • Catalyst activated water is known to enhance biological activities of organisms and enhances immunity when combined it with Bacillus species in drinking water, it improves the FCR and it probably had improved the birds’ resistance to diseases.
  • However, trials with more replicates are needed for a better conclusion.
 
March 6, 2018
Thankfully. ..so good explaning...
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Reply
Aurelio Tayao Aurelio Tayao
Veterinary Doctor
March 6, 2018
What antibiotic was used in the positive control group and why was the mortality much higher (13% vs 3%) in this treatment group than T1-T3? What was the cause of the mortalities?
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March 7, 2018
Aurelio Tayao
Dear Dr. Aurelio Tayao, Thank you for the interest shown in the technical article. A broad spectrum antibiotic containing fluroquinolone which inhibits DNA gyrase was used in the positive control group. The day old chicks used in the trial were culls. We followed the same procedure that normal broiler farmers here use, administration of prophylactic dose of antibiotic during first 3-5 days to control bacterial diseases (particularly E.coli & some times Salmonella gallinarum) coming from the hatcheries. Whatever the the weaknesses or infections that the chicks had, must have continued until the end of the trial causing mortality through out. Competitive extrusion, antibacterial compounds produced by the probiotic bacteria coupled with its immune stimulating properties must have contributed for controlling the moralities in treatment groups while CAW is known to reduce stress and improve immunity of farmed animals.
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Steffen Hansen Steffen Hansen
Animal Nutritionist
March 7, 2018
As I understand from the description of the trial design each treatment group had one replicate i.e. one pen per treatment? If this is the case, you cannot draw any conclusions at all. The results you have observed can be pure coincidence.
I am also missing a description of “catalyst activated water”.
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March 7, 2018
Steffen Hansen
Thank you Dr. Steffen Hansen for your comment. In my conclusions, the last one, I have mentioned that as the trial included only one replicate (31 chicks) per treatment, more trials should be done with at least four replicates for each treatment before a solid conclusion is made. The point that I wanted to make is that a mixture of spores of Bacillus species could work beneficially when administered via drinking water because some of the spores may germinate in the reservoir by the time (at least 4 hours) the birds drink making sure that the vegetative cells are consumed instead of spores. And also we have seen that 48 hours old Bacillus subtilis culture showing good inhibition of growth of some Gram negative bacteria (well diffusion method).. CAW is a patented product which has been invented by Dr. John Willard of USA. A synonym for this is Willard Water.
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Steffen Hansen Steffen Hansen
Animal Nutritionist
March 7, 2018
Dr. Christopher Hettiarachchi, thanks for elaborating on trial design. The application of probiotics in the drinking water is a promising concept. I wonder if you have made any thoughts on dosage; maybe a dose response trial? Bacillus products are usually added with 10^6 per gram feed. Another important issue is also if the addition of the probiotic product creates slime inside the water tubes.
In my opinion the concept of “catalyst activated water” is a scam. It is not possible to find any scientifically sound publications on this product. The suggested mode of actions on Dr. Willards homepage are also hilarious. I consider this “Dr. Willard water” in the same class as “Grander water” which is pure humbug.
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March 7, 2018
Steffen Hansen
Dr. Steffen Hansen I did not do a dose response trial. The bacterial density of the original solution was 2.44x107 CFU per ml, 100 ml of this was used for 1000 litres of drinking water. As you mentioned, there can be a build up of slime in the pipes & nipple drinkers which can hinder the water flow as the production cycle progresses but one has to do a trial to see how long it will take for slime to build up.
About CAW, I happened to read an interesting, lengthy article written by Dr. Willard on the invention of this product. I used this in Penaeus monodon hatchery together with a probiotic/bioaugmenter without antibiotic during larval rearing in 6, 5000 liter tanks each. It synchronized moulting. I tried CAW product for plant on seed germination. It gave higher % of germination. Though more replicates are needed, the present trial with CAW has produced some positive results.
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Kb Bohara Kb Bohara
Doctor of Veterinary Medicine
March 15, 2018
I fully agreed with Mr Steffen Hansen's comments here that the problem in front of our generation to be addressed is that we have to preserve nature's biodiversity in this planet,ways out to increase food production ( 30 to 70 percent by 2030 -2050)without violating animal welfares and rights, and be able to deliver a safe and quality food products in a affordable prices. So under such circumstances we got to learn just lesions from Hilarious class of research for example Gardener water(early 20 th century), Dr Willard water or CAW (1930s) and so called world changing class of research by Dr Ancel Keys (1950s) but do powerful research which could not be prove fake.
The point is in your nice research study's result you recorded 3 percent vs 13 percent mortality on treatment vs control groups which is very significant difference but recommendation is you need a properly designed study again ! The risk is that some cleaver company can take this unripen reference to make money -loss to our sustainable food production goal.
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March 16, 2018
Kb Bohara
Dear Dr. KB Bohara, Thank you for the comment. I am not interested in promoting CAW. I have nothing to do with this product or I have no connection with this company. What interested me in this product is the mode of action of its constituents on biological activities and that is why I included this in my field trial. The main point which I wanted to share with the community is that the mode of delivery of probiotic bacteria, particularly Bacillus species. The Bacillus species containing broiler feed contains only the spores because vegetative cells do not withstand pelleting process of feed manufacturing and the spores ingested through feed will take few hours for germination. But if we can deliver vegetative form of the probiotic bacterial cells with water, the effectiveness could be better. Also the administered bacteria can proliferate in the drinking water system with time displacing other undeniable bacteria such as E.coli. If this process last without blocking the drinking water system for 45 days, Hydrogen peroxide flush will remove the slime. Antibiotic free broiler rearing is a challenge unless feed millers and veterinary doctors are geared toward it.
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Christoph Hutter Christoph Hutter
Poultry farmer
ADDCON ADDCON
Porsgrunn, Germany
March 17, 2018
Hi

Very interesting to read this discussion.
I wonder if you did trials with adding acid in the water as well in the past.
There are new generation of liquid acid to be added and to keep the formation of slime and other things in pipe down.
And if you have a good mix some will end up in the destine of the animals with the water.
In case of Bacillus could be synergy between acid and Bacillus.
We are running trials with lactic acid in liquid pig feed systems as well in fermentation of feed.
So for poultry this could be interesting as well in the future a lot of scientist believe.
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Reply
March 19, 2018
Christoph Hutter
Hi Christop Huter
Yes I have used a commercial product containing a blend of organic acids in drinking water to curb bacterial infections in broiler rearing in evap. cooling houses.
The optimum pH for growth of most Bacillus species is close to neutral and therefore, addition of an organic acid like lactic acid will bring pH of drinking water to around 4 (recommended pH). It is known that activity of probiotic bacteria is manifested when they are physiological active (growing actively), Bacillus sp. in a medium of pH 4 would not grow actively and probiotic activity can not be expected under such situation except some enhanced immune activity due to bacterial cell wall components. As it was stated, it could be a good treatment for dislodging the climes in drinking watering tubes & nipples.
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Christoph Hutter Christoph Hutter
Poultry farmer
ADDCON ADDCON
Porsgrunn, Germany
March 19, 2018
Today i dont go anymore to 4,0 as the risk of less water intake and other things could be too much. You only need to go to 4.0 if you have acids in which some bacteria could se as feed like lactic acid. With an good blend to go to 4,4 - 4,5 it sould be enough to stopp nearly all reactions. Soem Bacillus should be still happy at this ph but depend on the kind you are right.
Maybe there are some other once we should use.
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April 18, 2018
Good article which every farmer should read
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