Lactobacilli are commonly used as probiotics in poultry to improve production parameters and to increase chicken resistance to enteric infections. However, lactobacilli do not efficiently colonise the chicken intestinal tract, and also, their anti-infection effect in vivo is sometimes questionable. In this study, we therefore evaluated the potential of a mixture of four Lactobacillus species (L. salivarius, L. reuteri, L. ingluviei and L. alvi) for the protection of chickens against Salmonella Enteritidis infection. Whenever the chickens were inoculated by lactobacilli and S. Enteritidis separately, there was no protective effect of lactobacilli. This means that when lactobacilli and S. Enteritidis are exposed to each other as late as in the crop of chickens, lactobacilli did not influence chicken resistance to S. Enteritidis at all. The only positive effect was recorded when the mixture of lactobacilli and S. Enteritidis was used for the inoculation of feed and the feed was anaerobically fermented for 1 to 5 days. In this case, chickens fed such a diet remained S. Enteritidis negative. In vitro experiments showed that the protective effect was caused by acidification of feed down to pH 4.6 due to lactobacilli fermentation and was associated with S. Enteritidis inactivation. The probiotic effect of lactobacilli was thus expressed in the feed, outside the chicken host.

Veterinary Research Institute 621 00 Brno Czech Republic

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Rychlik I. Composition and function of chicken gut microbiota. Animals. 2020;10:597. doi: 10.3390/ani10010103. PubMed DOI PMC

Zamojska D., Nowak A., Nowak I., Macierzynska-Piotrowska E. Probiotics and postbiotics as substitutes of antibiotics in farm animals: A Review. Animals. 2021;11:3431. doi: 10.3390/ani11123431. PubMed DOI PMC

Kollarcikova M., Kubasova T., Karasova D., Crhanova M., Cejkova D., Sisak F., Rychlik I. Use of 16S rRNA gene sequencing for prediction of new opportunistic pathogens in chicken ileal and cecal microbiota. Poult. Sci. 2019;98:2347–2353. doi: 10.3382/ps/pey594. PubMed DOI

Reid G., Gaudier E., Guarner F., Huffnagle G.B., Macklaim J.M., Munoz A.M., Martini M., Ringel-Kulka T., Sartor B., Unal R., et al. Prebiotics, Responders and non-responders to probiotic interventions: How can we improve the odds? Gut Microbes. 2010;1:200–204. doi: 10.4161/gmic.1.3.12013. PubMed DOI PMC

Cressman M.D., Yu Z., Nelson M.C., Moeller S.J., Lilburn M.S., Zerby H.N. Interrelations between the microbiotas in the litter and in the intestines of commercial broiler chickens. Appl. Environ. Microbiol. 2010;76:6572–6582. doi: 10.1128/AEM.00180-10. PubMed DOI PMC

De Cesare A., Caselli E., Lucchi A., Sala C., Parisi A., Manfreda G., Mazzacane S. Impact of a probiotic-based cleaning product on the microbiological profile of broiler litters and chicken caeca microbiota. Poult. Sci. 2019;98:3602–3610. doi: 10.3382/ps/pez148. PubMed DOI

Adams M.R. Safety of industrial lactic acid bacteria. J. Biotechnol. 1999;68:171–178. doi: 10.1016/S0168-1656(98)00198-9. PubMed DOI

Bongaerts G.P., Severijnen R.S. The beneficial, antimicrobial effect of probiotics. Med. Hypotheses. 2001;56:174–177. doi: 10.1054/mehy.2000.1135. PubMed DOI

Kubasova T., Seidlerova Z., Rychlik I. Ecological adaptations of gut microbiota members and their consequences for use as a new generation of probiotics. Int. J. Mol. Sci. 2021;22:5471. doi: 10.3390/ijms22115471. PubMed DOI PMC

Videnska P., Faldynova M., Juricova H., Babak V., Sisak F., Havlickova H., Rychlik I. Chicken faecal microbiota and disturbances induced by single or repeated therapy with tetracycline and streptomycin. BMC Vet. Res. 2013;9:30. doi: 10.1186/1746-6148-9-30. PubMed DOI PMC

Sengupta R., Altermann E., Anderson R.C., McNabb W.C., Moughan P.J., Roy N.C. The role of cell surface architecture of lactobacilli in host-microbe interactions in the gastrointestinal tract. Mediat. Inflamm. 2013;2013:237921. doi: 10.1155/2013/237921. PubMed DOI PMC

Bernardeau M., Guguen M., Vernoux J.P. Beneficial lactobacilli in food and feed: Long-term use, biodiversity and proposals for specific and realistic safety assessments. FEMS Microbiol. Rev. 2006;30:487–513. doi: 10.1111/j.1574-6976.2006.00020.x. PubMed DOI

Medvecky M., Cejkova D., Polansky O., Karasova D., Kubasova T., Cizek A., Rychlik I. Whole genome sequencing and function prediction of 133 gut anaerobes isolated from chicken caecum in pure cultures. BMC Genom. 2018;19:561. doi: 10.1186/s12864-018-4959-4. PubMed DOI PMC

Klaenhammer T.R. Get cultured: Eat bacteria. Annu. Rev. Food Sci. Technol. 2019;10:1–20. doi: 10.1146/annurev-food-032818-121826. PubMed DOI

Crhanova M., Karasova D., Juricova H., Matiasovicova J., Jahodarova E., Kubasova T., Seidlerova Z., Cizek A., Rychlik I. Systematic culturomics shows that half of chicken caecal microbiota members can be grown in vitro except for two lineages of Clostridiales and a single lineage of Bacteroidetes. Microorganisms. 2019;7:496. doi: 10.3390/microorganisms7110496. PubMed DOI PMC

Tellez G., Pixley C., Wolfenden R.E., Layton S.L., Hargis B.M. Probiotics/direct fed microbials for Salmonella control in poultry. Food Res. Int. 2012;45:628–633. doi: 10.1016/j.foodres.2011.03.047. DOI

Wang S., Peng Q., Jia H.M., Zeng X.F., Zhu J.L., Hou C.L., Liu X.T., Yang F.J., Qiao S.Y. Prevention of Escherichia coli infection in broiler chickens with Lactobacillus plantarum B1. Poult. Sci. 2017;96:2576–2586. doi: 10.3382/ps/pex061. PubMed DOI

Taha-Abdelaziz K., Astill J., Kulkarni R.R., Read L.R., Najarian A., Farber J.M., Sharif S. In vitro assessment of immunomodulatory and anti-Campylobacter activities of probiotic lactobacilli. Sci. Rep. 2019;9:17903. doi: 10.1038/s41598-019-54494-3. PubMed DOI PMC

Neal-McKinney J.M., Lu X., Duong T., Larson C.L., Call D.R., Shah D.H., Konkel M.E. Production of organic acids by probiotic lactobacilli can be used to reduce pathogen load in poultry. PLoS ONE. 2012;7:e43928. doi: 10.1371/journal.pone.0043928. PubMed DOI PMC

Kajander K., Hatakka K., Poussa T., Farkkila M., Korpela R. A probiotic mixture alleviates symptoms in irritable bowel syndrome patients: A controlled 6-month intervention. Aliment. Pharmacol. Ther. 2005;22:387–394. doi: 10.1111/j.1365-2036.2005.02579.x. PubMed DOI

Davis G.S., Anderson K.E. The effects of feeding the direct-fed microbial, primalac, on growth parameters and egg production in single comb white leghorn hens. Poult. Sci. 2002;81:755–759. doi: 10.1093/ps/81.6.755. PubMed DOI

Talebi A., Amirzadeh B., Mokhtari B., Gahri H. Effects of a multi-strain probiotic (PrimaLac) on performance and antibody responses to Newcastle disease virus and infectious bursal disease virus vaccination in broiler chickens. Avian Pathol. 2008;37:509–512. doi: 10.1080/03079450802356995. PubMed DOI

Kubasova T., Kollarcikova M., Crhanova M., Karasova D., Cejkova D., Sebkova A., Matiasovicova J., Faldynova M., Sisak F., Babak V., et al. Gut anaerobes capable of chicken caecum colonisation. Microorganisms. 2019;7:597. doi: 10.3390/microorganisms7120597. PubMed DOI PMC

Zheng J., Wittouck S., Salvetti E., Franz C., Harris H.M.B., Mattarelli P., O’Toole P.W., Pot B., Vandamme P., Walter J., et al. A taxonomic note on the genus Lactobacillus: Description of 23 novel genera, emended description of the genus Lactobacillus Beijerinck 1901, and union of Lactobacillaceae and Leuconostocaceae. Int. J. Syst. Evol. Microbiol. 2020;70:2782–2858. doi: 10.1099/ijsem.0.004107. PubMed DOI

Methner U., Barrow P.A., Gregorova D., Rychlik I. Intestinal colonisation-inhibition and virulence of Salmonella phoP, rpoS and ompC deletion mutants in chickens. Vet. Microbiol. 2004;98:37–43. doi: 10.1016/j.vetmic.2003.10.019. PubMed DOI

Kollarcikova M., Faldynova M., Matiasovicova J., Jahodarova E., Kubasova T., Seidlerova Z., Babak V., Videnska P., Cizek A., Rychlik I. Different Bacteroides species colonise human and chicken intestinal tract. Microorganisms. 2020;8:1483. doi: 10.3390/microorganisms8101483. PubMed DOI PMC

Busarcevic M., Dalgalarrondo M. Purification and genetic characterisation of the novel bacteriocin LS2 produced by the human oral strain Lactobacillus salivarius BGHO1. Int. J. Antimicrob. Agents. 2012;40:127–134. doi: 10.1016/j.ijantimicag.2012.04.011. PubMed DOI

Hai D., Huang X. Protective effect of Lactobacillus reuteri Lb11 from chicken intestinal tract against Salmonella Enteritidis SE05 in vitro. Antonie Van Leeuwenhoek. 2021;114:1745–1757. doi: 10.1007/s10482-021-01625-4. PubMed DOI

Evangelista A.G., Correa J.A.F., Dos Santos J.V.G., Matte E.H.C., Milek M.M., Biauki G.C., Costa L.B., Luciano F.B. Cell-free supernatants produced by lactic acid bacteria reduce Salmonella population in vitro. Microbiology. 2021;167:001102. doi: 10.1099/mic.0.001102. PubMed DOI

Okamoto A.S., Andreatti Filho R.L., Milbradt E.L., Moraes A.C.I., Vellano I.H.B., Guimaraes-Okamoto P.T.C. Bacterial communication between Lactobacillus spp. isolated from poultry in the inhibition of Salmonella Heidelberg-proof of concept. Poult. Sci. 2018;97:2708–2712. doi: 10.3382/ps/pey141. PubMed DOI

Huang J., Li J., Li Q., Li L., Zhu N., Xiong X., Li G. Peptidoglycan derived from Lactobacillus rhamnosus MLGA up-regulates the expression of chicken beta-defensin 9 without triggering an inflammatory response. Innate Immun. 2020;26:733–745. doi: 10.1177/1753425920949917. PubMed DOI PMC

Sun Z., Huang L., Kong J., Hu S., Zhang X., Kong W. In vitro evaluation of Lactobacillus crispatus K313 and K243: High-adhesion activity and anti-inflammatory effect on Salmonella Braenderup infected intestinal epithelial cell. Vet. Microbiol. 2012;159:212–220. doi: 10.1016/j.vetmic.2012.03.043. PubMed DOI

Menconi A., Wolfenden A.D., Shivaramaiah S., Terraes J.C., Urbano T., Kuttel J., Kremer C., Hargis B.M., Tellez G. Effect of lactic acid bacteria probiotic culture for the treatment of Salmonella enterica serovar Heidelberg in neonatal broiler chickens and turkey poults. Poult. Sci. 2011;90:561–565. doi: 10.3382/ps.2010-01220. PubMed DOI

Zhang D., Li R., Li J. Lactobacillus reuteri ATCC 55730 and L22 display probiotic potential in vitro and protect against Salmonella-induced pullorum disease in a chick model of infection. Res. Vet. Sci. 2012;93:366–373. doi: 10.1016/j.rvsc.2011.06.020. PubMed DOI

Penha Filho R.A., Diaz S.J., Fernando F.S., Chang Y.F., Andreatti Filho R.L., Berchieri A., Jr. Immunomodulatory activity and control of Salmonella Enteritidis colonization in the intestinal tract of chickens by Lactobacillus based probiotic. Vet. Immunol. Immunopathol. 2015;167:64–69. doi: 10.1016/j.vetimm.2015.06.006. PubMed DOI

Adhikari P., Lee C.H., Cosby D.E., Cox N.A., Kim W.K. Effect of probiotics on fecal excretion, colonization in internal organs and immune gene expression in the ileum of laying hens challenged with Salmonella Enteritidis. Poult. Sci. 2019;98:1235–1242. doi: 10.3382/ps/pey443. PubMed DOI

Yamawaki R.A., Milbradt E.L., Coppola M.P., Rodrigues J.C., Andreatti Filho R.L., Padovani C.R., Okamoto A.S. Effect of immersion and inoculation in ovo of Lactobacillus spp. in embryonated chicken eggs in the prevention of Salmonella Enteritidis after hatch. Poult. Sci. 2013;92:1560–1563. doi: 10.3382/ps.2012-02936. PubMed DOI

Vandeplas S., Dauphin R.D., Thiry C., Beckers Y., Welling G.W., Thonart P., Thewis A. Efficiency of a Lactobacillus plantarum-xylanase combination on growth performances, microflora populations, and nutrient digestibilities of broilers infected with Salmonella Typhimurium. Poult. Sci. 2009;88:1643–1654. doi: 10.3382/ps.2008-00479. PubMed DOI

Carter A., Adams M., La Ragione R.M., Woodward M.J. Colonisation of poultry by Salmonella Enteritidis S1400 is reduced by combined administration of Lactobacillus salivarius 59 and Enterococcus faecium PXN-33. Vet. Microbiol. 2017;199:100–107. doi: 10.1016/j.vetmic.2016.12.029. PubMed DOI

Higgins J.P., Higgins S.E., Wolfenden A.D., Henderson S.N., Torres-Rodriguez A., Vicente J.L., Hargis B.M., Tellez G. Effect of lactic acid bacteria probiotic culture treatment timing on Salmonella Enteritidis in neonatal broilers. Poult. Sci. 2010;89:243–247. doi: 10.3382/ps.2009-00436. PubMed DOI

Neveling D.P., van Emmenes L., Ahire J.J., Pieterse E., Smith C., Dicks L.M.T. Effect of a multi-species probiotic on the colonisation of Salmonella in broilers. Probiotics Antimicrob. Proteins. 2020;12:896–905. doi: 10.1007/s12602-019-09593-y. PubMed DOI

Markazi A., Luoma A., Shanmugasundaram R., Mohnl M., Raj Murugesan G., Selvaraj R. Effects of drinking water synbiotic supplementation in laying hens challenged with Salmonella. Poult. Sci. 2018;97:3510–3518. doi: 10.3382/ps/pey234. PubMed DOI

Smialek M., Kaczorek E., Szczucinska E., Burchardt S., Kowalczyk J., Tykalowski B., Koncicki A. Evaluation of Lactobacillus spp. and yeast based probiotic (Lavipan) supplementation for the reduction of Salmonella Enteritidis after infection of broiler chickens. Pol. J. Vet. Sci. 2019;22:5–10. doi: 10.24425/pjvs.2018.125616. PubMed DOI

Johnson T.J., Youmans B.P., Noll S., Cardona C., Evans N.P., Karnezos T.P., Ngunjiri J.M., Abundo M.C., Lee C.W. A consistent and predictable commercial broiler chicken bacterial microbiota in antibiotic-free production displays strong correlations with performance. Appl. Environ. Microbiol. 2018;84:e00362-18. doi: 10.1128/AEM.00362-18. PubMed DOI PMC

Wang L., Lilburn M., Yu Z. Intestinal microbiota of broiler chickens as affected by litter management regimens. Front. Microbiol. 2016;7:593. doi: 10.3389/fmicb.2016.00593. PubMed DOI PMC

Ngunjiri J.M., Taylor K.J.M., Abundo M.C., Jang H., Elaish M., Kc M., Ghorbani A., Wijeratne S., Weber B.P., Johnson T.J., et al. Farm stage, bird age, and body site dominantly affect the quantity, taxonomic composition, and dynamics of respiratory and gut microbiota of commercial layer chickens. Appl. Environ. Microbiol. 2019;85:e03137-18. doi: 10.1128/AEM.03137-18. PubMed DOI PMC

Ren H., Vahjen W., Dadi T., Saliu E.M., Boroojeni F.G., Zentek J. Synergistic effects of probiotics and phytobiotics on the intestinal microbiota in young broiler chicken. Microorganisms. 2019;7:684. doi: 10.3390/microorganisms7120684. PubMed DOI PMC

El-Sharkawy H., Tahoun A., Rizk A.M., Suzuki T., Elmonir W., Nassef E., Shukry M., Germoush M.O., Farrag F., Bin-Jumah M., et al. Evaluation of Bifidobacteria and Lactobacillus probiotics as alternative therapy for Salmonella typhimurium infection in broiler chickens. Animals. 2020;10:1023. doi: 10.3390/ani10061023. PubMed DOI PMC

Zhang L., Zhang R., Jia H., Zhu Z., Li H., Ma Y. Supplementation of probiotics in water beneficial growth performance, carcass traits, immune function, and antioxidant capacity in broiler chickens. Open Life Sci. 2021;16:311–322. doi: 10.1515/biol-2021-0031. PubMed DOI PMC

Beal R.K., Wigley P., Powers C., Hulme S.D., Barrow P.A., Smith A.L. Age at primary infection with Salmonella enterica serovar Typhimurium in the chicken influences persistence of infection and subsequent immunity to re-challenge. Vet. Immunol. Immunopathol. 2004;100:151–164. doi: 10.1016/j.vetimm.2004.04.005. PubMed DOI

Varmuzova K., Faldynova M., Elsheimer-Matulova M., Sebkova A., Polansky O., Havlickova H., Sisak F., Rychlik I. Immune protection of chickens conferred by a vaccine consisting of attenuated strains of Salmonella Enteritidis, Typhimurium and Infantis. Vet. Res. 2016;47:94. doi: 10.1186/s13567-016-0371-8. PubMed DOI PMC

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