Application of probiotics in commercial layer diets: a review

SD Chowdhury1, BC Ray1*, A Khatun1, MRA Redoy2 and AS Afsana1

1
Department of Poultry Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh;
2
Department of Animal Nutrition, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

Abstract
Probiotics of various commercial preparations are being considered worldwide for poultry as
performance enhancers and suitable alternatives to antibiotics. Much of the previous efforts to explore
beneficial effects of probiotics were directed for commercial broilers and scientific information for
commercial layer nutrition is limited in contrast to that of broilers. To our knowledge, whatever
information is available, no attempt has been made as yet to make a comprehensive review of the
effects of probiotics on layer chicken covering different aspects of production. This review is prepared by
consolidating and updating information available in the literature covering laying performance, serum
chemistry, egg quality and its safety aspects. Data generated from a number of studies clearly indicated
that apart from a few exceptions, probiotics are found to be effective for the improvement of laying
performance; serum biochemical parameters, egg shell and albumen quality and they are suitable for
the production of safe eggs. Effect of probiotics on layers is mediated by boosting up gut health thereby
creating an environment for better nutrient assimilation and thus an improvement of production and egg
quality is achieved. Factors affecting such results need to be assessed more precisely since they
contribute to some contradictions in the literatures. Nevertheless, the feed industry can rely on
probiotics as quite safe for the production of quality feed to support commercial layer nutrition.

Keywords: probiotics, laying performance, egg quality, serum chemistry, food safety

Bangladesh Animal Husbandry Association. All rights reserved. Bang. J. Anim. Sci. 2020. 49 (1):1-12

or harmful based on their response to the host

Introduction animal. Attachment of beneficial microflora is
Application of biotechnological tools to boost up always desired but negative to it may also occur.
the production efficiency of poultry is a dynamic The beneficial organisms maintain gut
feed technology. Such technology needs to be equilibrium, improve health of birds and enhance,
well justified so that it does not create any harm or at least maintain production. On the contrary,
to poultry as well as to the ultimate consumers of harmful bacteria like E. coli, Salmonella, Coliform
poultry products. With the advancement of and others may alter the gut equilibrium to a
biotechnology in poultry feeds and nutrition and negative direction. This problem can be
banning of harmful growth promoters, antibiotics elucidated by the supplementation of probiotics
in particular, the use of probiotics is gaining as they are believed to have a positive impact on
momentum globally in feed formulation strategy. poultry. Until recently, a number of beneficial
Antimicrobial resistance is now a worldwide effects have been reported from research works
anxiety (WHO, 2018) and alteration of immune with probiotics although there are some
response due to feeding of antibiotic growth controversies. Supplementation of commercial
promoters, has lead poultry nutritionists to find preparations of probiotics through diet comprising
out suitable alternatives to antibiotics. Probiotics either single or multi-strain positively affected
of various preparations are considered to be laying performance and egg quality
important tools in this regard (Kabir, 2007). It is characteristics specifically shell thickness and
now well established that newly hatched chicks’ Haugh unit of commercial layers (Ray, 2018). The
gut is sterile and establishment of microflora in factors on which exploration of beneficial effects
the gut of chicken starts just after hatch from of probiotics depend are the survival, stability
hatching tray and hatcher which is called “cloacal and strain of organisms, type of host animal,
drinking”, thereafter gradually with the manufacturing process, dose frequency, health
introduction of feed and water. Microorganisms and nutritional status of the birds, the age,
which colonize in the gut may be either beneficial physiological stress level, genetics of the host

*Corresponding author: bipul_ps@bau.edu.bd 1

 Probiotics in commercial layer diets

etc. (Chichlowski et al., 2007; Aalaei et el., spp. Among probiotic organisms, Saccharomyces
2018). Much of the research works with are known to offer a source of good quality
probiotics has been concentrated with broiler protein and B complex vitamins. Probiotics help in
birds and information with layer birds, whatever the digestion of food more efficiently, exert better
available, is not yet reported systematically. In metabolism, maintain the health status of the
addition, variations in the result due to birds and cause faster weight gain in chicken
application of different probiotics have been (Park et. al., 2016; Azemraw and Sewalem,
reported due to factors as mentioned earlier. 2017). Among probiotic bacteria, a small amount
Although improvement in egg production (Ribeiro of anaerobic bacteria is found in crop,
et al., 2014), health of birds and egg quality proventriculus and gizzard (Rastall, 2004) and
(Lokhande et al., 2013) have been reported, large amount of facultative anaerobic bacteria are
there are also inconsistencies in the literature. To available in small intestine (Khan and Naz, 2013).
authors’ knowledge, no attempt has been taken Probiotic bacteria colonize in enterocyte, caecal
yet to consolidate and update the results of and colonic epithelium, having the most colonized
feeding probiotics to layers in particular, which region, colon and caecum (1010-1013 cfu/g)
are available sporadically in the literatures. (Heczko et al., 2000). Probiotics are supplied
Therefore, this review is prepared in a through powder, in encapsulated form or liquid
synchronized form by consolidating and updating form either in feed or water. Their use seems to
the results of probiotics’ use in commercial layer be very much strain specific and no explicit
nutrition covering productive performance, serum recommendation has yet been developed for
chemistry, egg quality and safety aspects. particular strains and thus has deviations in their
feeding results. They deliberate beneficial effects
The concept of probiotics
by producing favorable conditions inside the gut
The word ‘probiotic’ was used by Elie Metchnikoff of poultry to enhance performances.
in 1906, a Russian scientist and he was awarded
Egg Production
Noble prize in medicine. He showed the beneficial
effects of microbes replacing those which were Supplementation of probiotics in the diet of laying
harmful for treating intestinal illness. He is known chicken favours egg production performance
as the father of probiotics (Rautray et al., 2011). either by increasing egg number or maintaining
The probiotics are considered as “direct-fed production. Some probiotic preparations are also
microbial” and they affect the host positively by effective in adverse or stressful conditions and
balancing the intestinal microbial populations supposed to have no detrimental effects when
(Fuller, 1989). Earlier, probiotics were referred to used in appropriate amount. Research findings
as substances which stimulate one another (Lilly showed that use of probiotics in layer diets
and Stillwell, 1965). Currently, the term is used enhanced egg production (Ribeiro et al., 2014;
for animal feed supplement (Saleh and Hayashi, Peralta-Sánchez et. al., 2019). Deviations to
2011) to explore beneficial effects. FAO and WHO these observations also exist, but in many cases
jointly defined probiotics as beneficial organisms it affects the production positively, or at least
having beneficial effects on the body of host maintains production as usual. Inclusion of
when administered in sufficient amount Lactobacillus acidophilus (109 cfu/kg feed)
(FAO/WHO, 2001). This definition was adopted by significantly enhanced the number of eggs
the International Scientific Association for (Gallazi et al., 2009). A combination of
Probiotics and Prebiotics (Hill et al., 2014). Lactobacillus spp. and Bacillus spp. as liquid
Probiotics may be classified as either bacteria probiotics mixed culture when fed to the ISA
(spore forming and non-spore forming), single or Brown layers showed an increase in egg
multi-strain, yeast, single species, multi-species, production (Raka et al., 2014). Abdelqader et al.
allochthonous and autochthonous (FAO, 2016). (2013a) showed that feeding of Bacillus subtilis
Microorganisms that are commonly used as (maximum 2.3×108 cfu/g) had significantly better
probiotics are Lactobacillus, Streptococcus, egg production compared to control at doses of
Enterococcus, Bacillus, Clostridium, 1g/kg feed and 0.5 g/kg feed for a duration of 10
Bifidobacterium species and E. coli while yeast weeks during late production period starting from
and fungus used as probiotics include 64 weeks. Results of a study by Khan et al.
Saccharomyces cerevisiae and Aspergillus oryzae (2011) depicted that addition of probiotics
(Fuller, 1989). Bacteria and yeasts are used as (2×109 cfu/g) to the layer diet improved egg
spores or as living microorganisms. Non- production. The probiotic organisms used in the
colonizing species are Saccharomyces cerevisiae trial were L. plantarum, L. bulgaricus, L.
and Bacillus spp. (spores) while colonizing acidophilus, L. rhamnosus, Bifidobacterium
species include Lactobacillus and Enterococcus bifidum, Streptococcus thermophilus,

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 Chowdhury et al. (2020) Bang. J. Anim. Sci. 49 (1):1-12

Enterococcus faecium, Aspergillus oryzae and (2009) found that the addition of probiotics had
Candida pinpolopesi. Similar positive effects on significant effect on egg weight but not on egg
egg production in layer breeder aging 24 weeks mass. Egg mass was increased by
also revealed by Panda et al. (2008) with the supplementation of 0.4% and 0.8% of live yeast
supplementation of 100 mg probiotics per kg diet (S. cerevisiae) as stated by Hassanein and
(L. sporogens, 6x108 spore/g product). Increased Soliman (2010). Average egg weight was not
egg production has been reported by probiotic influenced significantly by adding yeast to diets
feeding containing Enterococci based preparation (Nursoy et al., 2004). Feeding viable
(Arpasova et al., 2016); Enterococcus faecium Lactobacillus at 1100 mg per kg (4.4 ×107
(Zhang and Kim, 2013); heat-inactivated L. cfu/kg) was found to increase egg size (Nahashon
salivarius and B. subtilis (Zhang et al., 2012) and et al., 1996). According to Mahdavi et al. (2005),
with a mixture of probiotics containing several diet containing B. subtilis and B. licheniformis did
organisms (Kalavathy et al., 2005). Mazanko et not increase egg weight significantly. Recently,
al. (2017) found positive improvement in egg Martinez et al. (2018) conducted an experiment
production due to feeding of B. subtilis and B. using S. cerevisiae fermentation product at a
amyloliquefaciens. Youruk et al. (2004) found dose of 1.25g/kg where they found no differences
linear increase in egg production when supplied in egg weight and egg mass of Hy-Line layer.
with a number of organisms containing Fathi et al. (2018) found no significant
Lactobacillus spp., Bifidobacterium bifidum, differences when three different breeds were fed
Streptococcus salivarius, E. faecium, A. oryza and on three concentrations of dietary probiotics
C. pintolopesii. Enhancement of egg laying was containing 4×109 cfu/g of B. subtilis. Abdelkader
observed by Hassanein and Soliman (2010), upon et al. (2013) reported that egg weight was
feeding Saccharomyces cerevisiae at 0.4% and gradually increased for B. subtilis supplemented
0.8% as compared to control. The improvement groups. Khan and Naz (2013) conducted a trial
in egg production due to low level of yeast where they used commercial multi-strain
inclusion is in agreement with the result of probiotics with 2×109 cfu/g (L. plantarum, L.
Shivani et al. (2003) and Shareef and Al- bulgaricus, L. acidophilus, L. rhamnosus,
Dabbagh (2009) who observed higher percentage Bifidobacterium bifidum, Streptococcus
of egg production for hens fed yeast culture thermophilus, E. faecium, A. oryzae and C.
probiotics supplemented diets than the control pinpolopesi) and found significant increase in egg
hens. In contrast, no change in egg production weight and egg mass compared to negative
was also reported by Balevi et al. (2001). Fathi et control. Panda et al. (2008) found no significant
al. (2018) conducted experiment with three differences in egg weight when they fed L.
concentrations of dietary probiotic (0, 200 and sporogenes at 100mg (6×108 spore) per kg diet.
400 g/t feed) containing 4×109 cfu/g of B. subtilis
Feed intake and feed conversion ratio
involving three different breeds (White Leghorn,
Saudi black and Saudi brown). They found Results of feed consumption due to
statistically no significant difference in egg supplementation of probiotics in the diet of layers
production performance among dietary are not consistent. No significant effects on feed
treatments. Martinez et al. (2018) conducted an consumption have been reported in a number of
experiment using S. cerevisiae fermentation studies (Mahdavi et al., 2005; Ramasamy et al.,
product at a dose of 1.25g/kg where they found 2009; Fathi et al., 2018). Some other authors
no differences in egg production of Hy-Line (Yousefi and Karkoodi, 2007; Panda et al., 2008;
commercial layer. Abdelqader et al., 2013a) also revealed no
significant differences in the feed intake of layers
Egg weight and egg mass output
due to inclusion of probiotics in the diet.
A significant improvement in egg weight has been However, decrease in feed intake in the probiotics
reported by Ramasamy et al. (2009) due to treated groups was found in other studies (Yoruk
feeding Lactobacillus culture containing probiotic et al., 2004; Gallazzi et al., 2009). In contrast,
to laying hen throughout the laying cycle. The an increase in feed consumption was recorded in
highest egg weight was chalked out by Raka et laying hens fed on probiotics (Raka et al., 2014;
al. (2014) from the supplementation of liquid Zhang and Kim, 2014). A study by Balevi et al.
probiotics mixed culture (0.45% v/v) of 68.12g (2009) has shown improved FCR through the
compared to control of 65.22g. Haddadin et al. supplementation of a commercial probiotics in the
(1996) reported that egg weight was significantly basal diet consisting L. plantarum, Bifidobacteria
increased in Lactobacillus culture fed hens during bifidum, Streptococcus salivaries, E. faecium, A.
20-68 weeks of laying hen. Daneshyar et al. oryza and C. pintolopesiie in brown layers. A

3
 Probiotics in commercial layer diets

number of studies have shown that probiotics Blood cholesterol

improved feed conversion ratios in layer chickens
Recently, researchers have shown that dietary
(Panda et al., 2003; Yoruk et al., 2004; Panda et
supplementation of probiotics decrease
al., 2008; Gallazzi et al., 2009; Abdelqader et al.,
cholesterol and triglycerides in blood (Moataz et
2013a; Raka et al., 2014). Inatomi et al. (2016)
al., 2018; Kanani et al., 2018). Results of earlier
found that inclusion of probiotics mixture in layer
studies were also consistent with cholesterol
diet having B. mesentericus, C. butyricum, and S.
lowering effects of probiotics fed birds (Mohan et
faecalis significantly improved feed mass into egg
al., 1995; Abdulrahim et al., 1996). Reducing
mass. Contrary to this result, there was no
blood cholesterol level due to B. subtilis inclusion
significant difference in FCR between hens fed
has also been reported in layers (Sobczak and
probiotics supplemented diets and hens fed the
Kozłowski, 2015). Lactobacillus culture containing
control diet (Yousefi and Karkoodi, 2007; Fathi et
probiotics supplementation reduced serum low-
al., 2018). Feed conversion was improved for
density lipoprotein (LDL) cholesterol (Kalavathy
hens fed the probiotics diet containing S.
et al., 2003). A study by Zhang and Kim (2013)
cerevisiae at 1.25g/kg (Martinez et al., 2018).
showed that serum total cholesterol
Body weight, body weight gain and concentration was decreased by feeding hens
survivability with probiotics. In another study of Zhang et al.
(2012), a significant drop in total cholesterol
Probiotics organisms are found to affect body
content was evident. Amer and Khan (2012)
weight positively. Sobczak and Kozłowski (2015)
showed that the supplementation of probiotics (L.
indicated that dietary B. subtilis led to a
acidophilus, B. subtilis, S. cerevisiae and
significant increase in the final body weights and
Aspergillus oryzae) caused significant reduction in
weight gains of layer chickens (18-42 weeks).
serum cholesterol concentration after 6 weeks.
Neijat et al. (2019) found increased body weight
Similar reduction also stated by Chuke and
through feeding probiotics. Amer and Khan
Didacus (2015). Sohail et al. (2011) used a
(2012) showed that the supplementation of
commercial multi-strain probiotic with a minimum
probiotics (L. acidophilus, B. subtilis, S. cerevisiae
presence of 2×109 cfu/g (L. plantarum, L.
and A. oryzae) in desi chicken indicated
bulgaricus, L. acidophilus, L. rhamnosus,
significant increase in body weight gain after 6
Bifidobacterium bifidum, Streptococcus
weeks of experiment starting from 8 days of age
thermophilus, E. faecium, A. oryzae and C.
feeding probiotics (1.5×109cfu per ml). Body
pinpolopesi). The study has shown a reduction in
weight did not significantly differ when L.
serum cholesterol but HDL-cholesterol was
acidophilus was fed in two experiments after 34
significantly increased. Mansoub (2010) reported
and 61 weeks of age (Gallazzi et al., 2009).
that triglycerides and total cholesterol level of
Panda et al. (2008) observed no differences in
serum significantly decreased in groups
the weight gains (25-40 weeks) of hens fed diets
supplemented with probiotics containing L.
supplemented with probiotic bacteria L.
acidophilus and L. casei compared to control
sporogenes with 6×109 spores per gram. In a
group. Shirley et al. (2017) found that probiotics
study by Abdelqader et al. (2013a), the body
supplementation decreased (P<0.05) serum total
weights of laying hens were not affected by feed
cholesterol at 36 weeks of age, and serum LDL
additives such as B. subtilis, inulin or synbiotics.
cholesterol, alanine aminotransferase and
Yousefi and Karkoodi (2007) found that body
alkaline phosphatase at 36 and 52 weeks of age.
weight changes did not differ significantly among
On the contrary, the dietary probiotics
treatment groups of probiotics (S. cerevisiae) at
supplementation did not affect serum cholesterol
doses of 0.05, 0.1 and 0.15% dietary
or triglycerides in some other studies (Zarie et
supplementation for 63-75 weeks in commercial
al., 2011; Mohebbifar et al., 2013).
layers. Body weight of layer was not significantly
affected by probiotics (L. acidophilus, L. casei, B. Egg yolk cholesterol
bi¢dum, A. oryzae, S. faceium and Torulopsis
Yolk cholesterol, triglycerides and very low
spp) during 25-72 weeks (Panda et al., 2003). A
density lipoprotein (VLDL) were significantly
decrease in mortality was found by Yoruk et al.
declined as a result of supplementation of B.
(2004) when probiotics was supplemented at a
subtilis and B. licheniformis compared to control
rate of 0.1% and 0.2% in the diet of late laying
or antibiotic-treated groups (Yang et al., 2020).
hens. The probiotic organisms were Lactobacillus,
Some other findings are also in line with the
Bifidobacterium, Streptococcus, and Enterococcus
cholesterol lowering effect of probiotics fed birds
spp.
(Kanani et al., 2018 and Ashayerizadeh et al.,
2011). The cholesterol content of eggs produced
by probiotic (Lactobacillus culture) fed hens was

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 Chowdhury et al. (2020) Bang. J. Anim. Sci. 49 (1):1-12

significantly reduced by 15.3% and 10.4% when the study. According to Aghaei et al. (2010) and
compared to those of the control hens at 24 and Mikulski et al. (2012), probiotics exerted a
28 weeks of age, respectively (Ramasamy et al., beneficial influence on eggshell thickness.
2009). Mikulski et al. (2012) reported that Contrary to a number of positive reports on shell
dietary supplementation of probiotics quality, Haddadin et al. (1996) found no
(Pediococcus acidilactici) decreased the egg significant effect (P>0.05) on shell hardness and
cholesterol content and fatty acid composition in shell thickness due to addition of a probiotic (L.
laying hens. Mousa et al. (2015) stated that egg acidophilus) in the diet. Fathi et al. (2018) found
yolk cholesterol was decreased markedly by that B. subtilis at 200 and 400g/ton feed
probiotics supplementation. Several other significantly increased shell weight.
authors also reported similar results (Panda et
Albumen quality
al., 2003; Panda et al., 2008; Ramasamy et al.,
2009) from their experiments where they found a An increase in Haugh unit has been recorded by
reduction of cholesterol concentration in egg yolk Gallazzi et al. (2009) with the inclusion of a
due to use of probiotics. In a study of Kurtoglu et probiotic containing L. acidophilus in the diet of
al. (2004), probiotics supplementation at 250, laying hen at a dose of 109 cfu/kg of feed. Eggs
500 and 750 mg/kg feed decreased the egg yolk laid by probiotics treated hens showed
cholesterol levels when compared with the significantly higher albumen quality in terms of
control. Mahdavi et al. (2005) also expressed Haugh unit value. Chung et al. (2015) conducted
cholesterol lowering effects of probiotics. Sohail an experiment with mixed probiotics containing
et al. (2011) conducted a study where they used five microbial species that included A. oryzae, B.
a total of 180 Hy-Line W-98 hens, 40 weeks old subtilis, S. cerevisiae, L. plantarum and
and fed a commercial multi-strain probiotic. In Rhodopseudomonas capsulate, and found that
that study, supplementation of a probiotic supplementation of 0.4% mixed probiotics
containing L. plantarum, L. bulgaricus, L. showed greatest (P<0.05) Haugh unit. Similar
acidophilus, L. rhamnosus, Bifidobacterium results have been revealed by Zhang et al.
bifidum, Streptococcus thermophilus, E. faecium, (2012) and Sobczak and Kozłowski (2015).
A. oryzae and C. pinpolopesi in laying hen diets Improvement of protein quality in terms of
has shown a reduction in egg yolk cholesterol albumen height and Haugh unit value due to
concentration. dietary supplementation of B. subtilis containing
probiotics has been noted in a recent study of
Egg shell quality
Neijat et al. (2019). Khan et al. (2011) found that
The dietary supplementation of probiotics Haugh unit score was significantly increased
improved shell quality and bone strength in (P<0.05) in the groups fed the multi-strain
laying hens (Yan et al., 2019). Increase in shell probiotics containing diets as compared to
thickness has been stated by researchers through negative control group. In contrast, no significant
probiotics supplementation (Panda et. al., 2003; enhancement of Haugh unit value was observed
Panda et al., 2008; Mohan et al., 1995; Ray, in several other studies (Mahdavi et al., 2005;
2018). Supplementation of probiotics tends to Panda et al., 2008; Mikulski et al., 2012; Fathi et
increase shell hardness and proportion positively. al., 2018).
This beneficial effect may be attributed to a
Yolk quality
favorable environment in the gut (Panda et al.,
2008; Mikulski et al., 2012) probably by calcium Improvement in the production of darker yolk
mobilization as a primary means to improve color to get higher yolk color score has been
eggshell quality (Skrivan et al., 2010). Li et al. reported in a number of experiments (Li et al.,
(2006) found that dried B. subtilis cultures 2006; Mikulski et al., 2012). Sobczak and
increased eggshell thickness. Balevi et al. (2009) Kozłowski (2015) showed that eggs laid by
reported that feeding with 0.5 g/kg multi strain probiotics treated group hens received
probiotics diet caused statistically significant significantly higher scores for yolk color. On the
reduction on damaged egg ratio. Sobczak and contrary, there was no significant difference in
Kozłowski (2015) found that probiotics for laying yolk color among treatments upon feeding mixed
hens contributed to an improvement in eggshell probiotics containing five microbial species
quality. Similar results were reported by (Chung et al., 2015). Besides, Xu et al. (2006)
Abdelqader et al. (2013 a,b), where they stated and Zhang et al. (2012) did not note any change
that supplementation of 1g/kg feed and 0.5g/kg in yolk color in response to feeding probiotic
feed exhibited 8.4% and 7.5% increase, bacteria. Yang et al. (2003) observed a quadratic
respectively, in eggshell thickness at the end of effect of graded concentration of supplemental

5
 Probiotics in commercial layer diets

probiotics on yolk color. Fathi et al. (2018) Discussion

reported no significant differences between the
dietary groups with respect to yolk per cent and Research reports reviewed for the preparation of
yolk color resulting from the supplementation of this review showed a lot of positive effects of
B. subtilis containing probiotic at different doses. probiotics on laying chicken even though some
No change in yolk color has been postulated in controversies still exist. Supplementation of
some other findings as well (Berrin 2011; Khan et probiotics enhances performance of the birds by
al. 2011). boosting gut health, better nutrient assimilation
and thus improvement in production is achieved.
Safety aspects of probiotics The general mode of action of probiotics for
Safe food production is a global demand getting advantageous effects may be direct
nowadays. Table egg, a cheapest source of antagonistic effects against particular organisms,
animal protein for human consumption must be resulting in decrease in number of harmful
pathogen free to satisfy the safety aspects. organisms or by an effect on their metabolism or
Indiscriminate use of antibiotic growth promoters by synthesis of some essential nutrients or by
in poultry to improve their productivity causes stimulation of immunity (Jadhav et al., 2015).
antimicrobial resistance to health of people But it is possible that more than one mechanism
following consumption of foods (egg/meat) from may work simultaneously. Probiotics reduce
chicken treated with that growth promoter (Park viable count through the production of
et al., 2016). Multi drug resistance organisms antibacterial substances (lactocidin, acidophillin,
(MDRO) or “Superbug” has already been emerged organic acids, bacteriocins) and hydrogen
both for animal and human due to continuous use peroxide. Due to natural breakdown of probiotics
of antibiotic growth promoters in the diet of and metabolism of nutrients in the
poultry. However, Alfredson and Korolik, (2007) gastrointestinal tract, the produced volatile fatty
reported an increase in prevalence of infectious acids and organic acids lower the pH below
disease by Campylobacter jejuni or Clostridium required for the survival of pathogenic bacteria
perfringens in birds, if antibiotic growth such as E. coli and Salmonella spp (Khan and
promoters are excluded from diet. In this Naz, 2013). As a result, the growth of harmful
situation, probiotics could be an alternative for bacteria is inhibited. Another mechanism of
growth promotion as well as to ensure food action is “competitive exclusion” by which
safety (Chowdhury, 2018). Most concentrated probiotic bacteria colonize in the adhesion sites
source of Salmonella spp. is the ceca of GIT in on the intestinal epithelium and prevent colony
poultry which subsequently contributed to the formation of pathogenic bacteria (Chichlowski et
contamination of eggs. Feeding of probiotics was al., 2007). They improve digestion by increased
found to be effective in reducing harmful bacterial activity of the digestive enzymes and better
population in the gut (Chichlowski et al., 2007). absorption of nutrients while decreased bacterial
Feeding probiotics containing Lactobacillus and enzyme activity (glucoronidase, nitroreductase,
Bifidobacterium spp. is related with the reduction azoreductase) which are produced by some
of Salmonella spp. (Cox and pavic, 2009). Among pathogenic bacteria. In addition, probiotics
the food pathogens, Salmonella, E. coli and reduce ammonia production. They stimulate
Campylobacter are significantly responsible for immune system by higher production of
contamination of food. A study from Hassanein immunoglobulins and increase activities of
and Soliman (2010) confirmed that feeding S. macrophages and lymphocytes and also by
cerevisiae in live form at doses of 0.4% and 0.8% amplification of the production of γ-interferon
reduced these three organisms along with (Yang and Choct, 2009). Moreover, probiotics
Klebsiella spp., Staphylococcus spp. and improve the intestinal morphology through
Micrococcus spp. Higgins et al. (2007) reported increasing the villus height, goblet cell number
that probiotics significantly increased and decrease the crypt depth to create the
Lactobacillus and decreased E. coli, Clostridium environment favourable to the host. In precise,
perfringens and Salmonella count in the lower gut they have the potentiality to create favorable
in poultry as well as in its excreta. Latorre et al. conditions in the host animal to boost up
(2019) noted that supplementation of Bacillus production. Out of many factors, the effect of
spp. (B. subtilis and B. amyloliquefaciens) in probiotics on poultry performance may differ
poultry diet reduced undesirable bacteria in the based on single or multi-strain composition and
duodenal content compared to control. Thus, use their doses (Ray, 2018). In case of single strain
of probiotics is seemed to be quite appropriate for probiotics, it is easy to clarify the exact
ensuring food safety during the course of mechanism of action but for multi-strain, it has
husbandry practices. become quite difficult to state precisely which
strain has actually contributed to what extent and

6
 Chowdhury et al. (2020) Bang. J. Anim. Sci. 49 (1):1-12

by which mechanisms. But, for multi-strain Ahrens et al., 2007). Reduced pH of intestinal
probiotics, effectiveness is achieved when there is lumen increases calcium solubility for absorption
synergistic action among different strains and in (Van den Heuvel et al., 1999) whereas epithelial
such condition, it is supposed that multi-strain cell proliferation and villus height are stimulated
probiotics have more adhesive power than single by the production of short chain fatty acids
strain (Timmerman et al., 2004). Studies (Garcia et al., 2007) to increase the absorption
revealed that probiotics increase egg production. rate (Scholz-Ahrens et al., 2007), thus improve
Such an increase in egg production might be due the egg shell quality. Such improvement in shell
to elongated small and large intestinal lengths, quality is more important for transportation and
suppressing effects of undesirable bacteria, marketing to reduce egg breakage and number of
stimulating effects on the growth of beneficial abnormal shaped eggs. This attribute is much
bacteria or enhanced activity of beneficial more helpful in aged hens where quality of shell
bacteria in the intestines, thereby triggering is a major concern. Stress conditions are harmful
absorptive capacity of nutrients (Chen et al. for birds’ performance in which probiotics are
2005) that could be accounted their increased good performers for maintaining production by
nutrients availability at the site of formation of balancing gut microflora. Good quality and safe
egg components. But, as already mentioned, eggs are possible to obtain in probiotic fed birds.
such results of improvement in egg laying An improvement in yolk color has been stated in
capacity are not always consistent. It appears a very few studies. A reasonable explanation for
that application of probiotics either enhance an improvement in yolk color might be due to the
production and egg quality or at least maintain fact that carotenoids from the diet is well
such parameters without showing any absorbed and transferred into the egg yolk more
detrimental effects. However, several researchers efficiently, although exact mechanism of such
reported a minimum concentrations of probiotics action is not definitely known. The survivability of
in each gram should be 106 cfu/g and for birds is always satisfactory because probiotics
effectiveness in the body, an animal should have boost up immunity and thus beneficial in the
a daily intake of 108-109 organisms (Patterson maintenance of birds’ health. Although
and Burkholder, 2003; Toma and Pokrotnieks, considerable evidences have been growing in the
2006), but the type of organisms, their dose and recent years that egg yolk cholesterol has no
length of feeding have not been explained as yet. influence on the cardiovascular problems
In many cases, probiotics are performance (Abdollahi et al., 2019; Rong et al., 2013; Shin et
enhancers as they improve efficiency in feed al., 2013), rather is useful in preventing heart
utilization and therefore improve feed conversion. diseases, researchers all over the world are
The reduced feed intake in probiotic fed birds making efforts to reduce cholesterol in eggs by
might be a reflection of efficient utilization of genetic (Elkin, 2007) and dietary manipulations
nutrients. Improving feed efficiency using (Chowdhury et al., 2002; Chowdhury et al.,
nutritional approaches are common phenomena. 2005; Dey et al., 2012; Mattioli et al., 2016). Use
Probiotics, being useful in efficient utilization of of probiotics to reduce cholesterol and other
feed by making the favorable environmental components (triglycerides, very low density
conditions inside the gut, improve feed lipoprotein) in eggs is a new dimension side by
conversion. Moreover, it added extra profit to the side with other dietary means of manipulation.
farm owners. In laying hen, growth is rarely According to Corcoran el al. (2005), when
affected during laying period as the feed is digestion of fat is concerned, it depends on the
utilized mostly for egg production rather than live level of gallbladder acids in digestion latex which
weight gain. Consequently, improvement in body subsequently contribute to the lipid content.
weight gain is noticeable in earlier phases of Probiotic organisms have the competency to
layers but maintenance of good health and reduce fat content in blood. As the fat content is
standard body weight in laying periods are always reduced in blood, it is generally believed that
important. Although a variety of factors are transportation to egg yolk would be less. Due to
responsible for having favourable effect of recent trends in exclusion of antibiotics in feed
probiotics on bird, they are supposed to affect the industry, probiotics appeared as a viable
host positively. Their introduction to poultry feed alternative not only as performance enhancers
improve egg quality, particularly shell thickness but also it plays an important role in ensuring the
and therefore, shell quality. The increase in shell production of safe eggs for human race. Unlike
thickness is associated with the production of some other non-nutritive growth promoters,
short chain fatty acids due to fermentation which probiotics are always considered to be safe as
subsequently reduces the luminal pH (Scholz-

7
 Probiotics in commercial layer diets

they leave no residues in egg for human intestinal flora. Journal of Animal and Veterinary
consumption. Advances 9:1996–2000.
Alfredson DA, and V Korolik (2007). Antibiotic
Conclusion resistance and resistance mechanisms in
Campylobacter jejuni and Campylobacter coli.
Dietary inclusion of probiotics for commercial FEMS Microbiology Letters 277:123–132.
layers may improve laying performances, and Amer MY and SH Khan (2012). A comparison
egg quality in terms of albumen and shell quality. between the effects of a probiotic and an
Application of probiotics improves serum antibiotic on the performance of Desi chickens.
Veterinary World 5(3):160–165.
biochemical parameters by reducing total
cholesterol, LDL-cholesterol and triglycerides, and Arpasova H, M Kačániová, V Pistová and L Hleba
increasing HDL-cholesterol. Factors affecting such (2016). Effect of probiotics and humic acid on
egg production and quality parameters of laying
results need to be assessed more precisely since
hens eggs. Animal Science and Biotechnologies
they contribute to some contradictions in the 49(2):1-2.
literature. Nevertheless, supplementation of
Ashayerizadeh A, N Dabiri, KH Mirzadeh and MR
probiotics in the diet of laying chicken is safe and
Ghorbani (2011). Effect of dietary
therefore may play a vital role globally not only supplementation of probiotic and prebiotic on
as performance enhancers but also as a means of growth indices and serum biochemical
producing safe eggs for human. parameters of broiler chickens. Journal of Cell
and Animal Biology 5:152-156.

Conflict of interest Azemraw W and M Sewalem (2017). Review on

Application of Probiotics in Poultry Production.
The authors have no conflict of interest to British Journal of Poultry Sciences 6(3):46-52.
declare. Balevi T, US Ucan, B Coskun, V Kurtoglu and
Cetingul S 2009: Effect of dietary probiotic on
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