In this study, Gram positive and catalase negative rod-shaped bacterial strains were isolated from Iran’s backyard chicken intestines. After a preliminary screening for acid and bile tolerance, 14 Lactobacillus reuteri isolates were selected based on susceptibility to the antibiotics and the absence of beta haemolysis for antimicrobial activity against Salmonella Enteritidis and Salmonella Typhimurium. All 14 isolates were able to tolerate pH 3 for 3 h and their resistance to 0.3% bile salts was more than 50%. Among these 14 isolates, L. reuteri ABRIG22, L. reuteri ABRIG3, L. reuteri ABRIG18, L. reuteri ABRIG17, L. reuteri ABRIG8 and L. reuteri ABRIG9 were able to withstand pH 2.3 for 2 h, and L. reuteri ABRIG17 was the only isolate with good survivability in pH 2.3 for 3 h. Five isolates were able to withstand stronger acidic conditions and inhibit Salmonella Enteritidis and Salmonella Typhimurium by more than 90% with less of their supernatants. The results of this study demonstrated that isolated L. reuteri from intestines of chickens native to Northern Iran could be introduced as potential antimicrobial probiotic strains to control S. Enteritidis and S. Typhimurium infection.
Bu çalışmada, İran’daki köy tavuklarının barsaklarından Gram pozitif ve katalaz negatif çomak şekilli bakteriler izole edildi. Asit ve safra toleransı için bir ön taramanın ardından 14 Lactobacillus reuteri izolatı antibiyotik direnci ile Salmonella Enteritidis ve Salmonella Typhimurium’a karşı antimikrobiyal aktivite için beta hemolizis şekillenmemesine göre seçildi. 14 izolatın tümü 3 saat süreyle pH 3’ü tolere edebildi ve %0.3 safra tuzlarına karşı direnci %50’den fazla idi. Bu 14 izolat içinde, L. reuteri ABRIG22, L. reuteri ABRIG3, L. reuteri ABRIG18, L. reuteri ABRIG17, L. reuteri ABRIG8 ve L. reuteri ABRIG9 2 saat süreyle pH 2.3’e dayanabildi. L. reuteri ABRIG17 3 saat süreyle pH 2.3’te hayatta kalabilen tek izolattı. Beş izolat daha sert asidik ortama dayanabildi. Bunlar ayrıca daha az supernatant ile Salmonella Enteritidis ve Salmonella Typhimurium’u %90’dan daha fazla inhibe edebildi. Bu çalışmanın sonuçları, Kuzay İran’da yerel tavukların barsaklarından izole edilen L. reuteri’nin, S. Enteritidis ve S. Typhimurium enfeksiyonlarında potansiyel antimikrobiyal probiyotikler olarak kullanılabileceğini göstermektedir.
___
Van Coillie E, Goris J, Cleenwerck I, Grijspeerdt K, Botteldoorn N, Van Immerseel F, De Buck J, Vancanneyt M, Swings J, Herman L, Heyndrickx M: Identification of lactobacilli isolated from the cloaca and vagina of laying hens and characterization for potential use as probiotics to control Salmonella Enteritidis. J Appl Microbiol, 102, 1095-1106, 2007. DOI: 10.1111/j.1365-2672.2006.03164.x
Casey PG, Casey GD, Gardiner GE, Tangney M, Stanton C, Ross RP, Hill C, Fitzgerald GF: Isolation and characterization of antiSalmonella lactic acid bacteria from the porcine gastrointestinal tract. Lett Appl Microbiol, 39, 431-438, 2004. DOI: 10.1111/j.1472-765X.2004.01603.x
Musikasang H, Tani A, H-kittikun A, Maneerat S: Probiotic potential of lactic acid bacteria isolated from chicken gastrointestinal digestive tract. World J Microbiol Biotechnol, 25, 1337-1345, 2009.
Jin LZ, HoYW, Abdullah N, Ali MA, Jalaludin S: Note: Lack of influence of adherent Lactobacillus isolates on the attachment of Escherichia coli to the intestinal epithelial cells of chicken in vitro. J Appl Microbiol, 84, 1171-1174, 1998. DOI: 10.1046/j.1365-2672.1998.00434.x
Garriga M, Pascual M, Monfort JM, Hugas M: Selection of lactobacilli for chicken probiotic adjuncts. J App Microbiol, 84, 125-132, 1998. DOI: 10.1046/j.1365-2672.1997.00329.x
Shokryazdan P, Sieo CC, Kalavathy R, Liang JB, Alitheen NB, Faseleh Jahromi M, Ho YW: Probiotic potential of Lactobacillus strains with antimicrobial activity against some human pathogenic strains. Biomed Res Int, 2014 (2014), Article ID: 927268, 2014. DOI: 10.1155/ 2014/927268
Taheri HR, Moravej H, Tabandeh F, Zaghari M, Shivazad M: Screening of lactic acid bacteria toward their selection as a source of chicken probiotic. Poult Sci, 88, 1586-1593, 2009. DOI: 10.3382/ps.2009- 00041
Sahadeva RPK, Leong SF, Chua KH, Tan CH, Chan HY, Tong EV, Wong SYW, Chan HK: Survival of commercial probiotic strains to pH and bile. Int Food Res J, 18, 1515-1522, 2011.
Gilliland SE, Staley TE, Bush LJ: Importance of bile tolerance of Lactobacillus acidophilus used as a dietary adjunct. J Dairy Sci, 67, 3045- 3051, 1984.
Shokryazdan P, Liang JB, Abdullah N, Faseleh Jahromi M: Probiotic potential of lactic acid bacteria isolated from mulberry silage. J Pure Appl Microbiol, 9, 443-452, 2015. DOI: 10.7324/JAPS.2013.30317
Maragkoudakis PA, Zoumpopoulou G, Miaris C, Kalantzopoulos G, Pot B, Tsakalidou E: Probiotic potential of Lactobacillus strains isolated from dairy products. Int Dairy J, 16, 189-199, 2006. DOI: 10.1016/j. idairyj.2005.02.009
Jin LZ, Ho YW, Abdullah N, Ali MA, Jalaludin S: Antagonistic effects of intestinal Lactobacillus isolates on pathogens of chicken. Lett Appl Microbiol, 23, 67-71, 1996. DOI: 10.1111/j.1472-765X.1996.tb00032.x
Bian L, Molan AL, Maddox I, Shu Q: Antimicrobial activity of Lactobacillus reuteri DPC16 supernatants against selected food borne pathogens. World J Microbiol Biotechnol, 27, 991-998, 2011. DOI: 10.1007/ s11274-010-0543-z
Touré R, Kheadr E, Lacroix C, Moroni O, Fliss I: Production of antibacterial substances by bifidobacterial isolates rom infant stool active against Listeria monocytogenes. J Appl Microbiol, 95, 1058-1069, 2003. DOI: 10.1046/j.1365-2672.2003.02085.x
Lane DJ: 16S/23S rRNA sequencing. In, Stackebrandt E, Goodfellow M (Eds): Nucleic Acid Techniques in Bacterial Systematics. 115-175, John Wiley and Sons, New York, 1991.
McDonald R, Kenna EM, Murrell JC: Detection of methanotrophic bacteria in environmental samples with the PCR. Appl Environ Microbiol, 61, 116-121, 1995.
Yamazaki M, Ohtsu H, Yakabe Y, Kishima M, Abe H: In vitro screening of lactobacilli isolated from chicken excreta to control Salmonella Enteritidis and Typhimurium. Br Poult Sci, 53, 183-189, 2012. DOI: 10.1080/00071668.2012.678814
Kumar A, Kumar D: Characterization of Lactobacillus isolated from dairy samples for probiotic properties. Anaerobe, 33, 117- 123, 2015. DOI:10.1016/j.anaerobe.2015.03.004
Ehrmann MA, Karzak P, Bauer J, Vogel RF: Characterization of lactobacilli towards their use as probiotic adjuncts in poultry. J Appl Microbiol, 92, 966-975, 2002. DOI: 10.1046/j.1365-2672.2002.01608.x
EFSA: Panel on additives and products or substances used in animal feed, guidance on the assessment of bacterial susceptibility to antimicrobials of human and veterinary importance. EFSA J, 10, 1-10, 2012.
Maragkoudakis PA, Mountzouris KC, Psyrras D, Cremonese S, Fischer J, Cantor MD, Tsakalidou E: Functional properties of novel protective lactic acid bacteria and application in raw chicken meat against Listeria monocytogenes and Salmonella Enteritidis. Int J Food Microbiol, 130, 219-226, 2009. DOI: 10.1016/j.ijfoodmicro.2009.01.027
Tellez G, Pixley C, Wolfenden RW, Layton SL, Hargis BM: Probiotics/ direct fed microbials for Salmonella control in poultry. Food Res Int, 45, 628-633, 2012. Doi: 10.1016/j.foodres.2011.03.047
Wang JY, Tang P, Cui EH, Wang LQ, Liu WH, Ren JJ, Wu N, Qiu YH, Liu HJ: Characterization of antimicrobial resistance and related resistance genes in Escherichia coli strains isolated from chickens in China during 2007-2012. Afr J Microbiol Res, 7 (46): 5238-5247, 2013. DOI: 10.5897/ AJMR2013.5860
Sodagari HR, Mashak Z, Ghadimianazar A: Prevalence and antimicrobial resistance of Salmonella serotypes isolated from retail chicken meat and giblets in Iran. J Infect Dev Ctries, 9, 463-469, 2015. DOI: 10.3855/ jidc.5945
Firoozeh F, Shahcheraghi F, Zahraei Salehi T, Karimi V, Aslani MM: Antimicrobial resistance profile and presence of class I integrongs among Salmonella enterica serovars isolated from human clinical specimens in Tehran, Iran. Iran J Microbiol, 3, 112-117, 2011.
Tajbakhsh M, Hendriksen RS, Nochi Z, Zali MR, Aarestrup FM, Garcia-Migura LL: Antimicrobial resistance in Salmonella spp. recovered from patients admitted to six different hospitals in Tehran, Iran from 2007 to 2008. Folia Microbiol (Praha), 57, 91-97, 2012. DOI: 10.1007/s12223-012- 0099-4