Abstract
The sialidase activity and genetic diversity of 22 Clostridium perfringens strains isolated from chickens with necrotic enteritis were determined. Sialidase activity was detected in 86.4 % of the strains. All C. perfringens showed a high value of similarity (>96 %), and they were grouped into seven clusters clearly separated from the other reference bacterial strains. From these clusters four patterns were defined in accordance with their phenotypic (sialidase production and antibiotic resistance profile) and genotypic (presence of nanI and nanJ genes) characteristics. Our results showed heterogeneity among strains, but they were genotypically similar, and it is suggested further studies are needed to better understand the pathogenesis of necrotic enteritis.
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Baker A, Davis E, Rehberger T, Rosener D (2010) Prevalence and diversity of toxigenic Clostridium perfringens and Clostridium difficile among swine herds in the midwest. Appl Environ Microbiol 76:2961–2967
Baums CG, Schotte U, Amtsberg G, Goethe R (2004) Diagnostic multiplex PCR for toxin genotyping of Clostridium perfringens isolates. Vet Microbiol 100:11–16
Caetano-Anollés G (1993) Amplifying DNA with arbitrary oligonucleotide primers. Genome Res 3:85–94
Chiarezza M, Lyras D, Pidot S, Flores-Díaz M, Awad M, Kennedy C, Cordner L, Phumoonna T, Poon R, Hughes M, Emmins J, Alape-Girón A, Rood J (2009) The nanI and nanJ sialidases of Clostridium perfringens are not essential for virulence. Infect Immun 77:4421–4428
Clinical and Laboratory Standards Institute (CLSI). 2007. Methods for antimicrobial susceptibility testing of anaerobic bacteria, seventh edn. Wayne, (M11-A7)
Coursodon CF, Glock RD, More KL, Cooper KK, Songer JG (2012) TpeL-producing strains of Clostridium perfringens type A are highly virulent for broiler chicks. Anaerobe 18(1):117–121
Engstrom B, Johansson A, Aspan A, Kaldhusdal M (2012) Genetic relatedness and netB prevalence among environmental Clostridium perfringens strains associated with a broiler flock affected by mild necrotic enteritis. Vet Microbiol 59:260–264
Eribe ER, Olsen I (2000) Strain differentiation in Bacteroides fragilis by RAPD and dendron computer-assisted gel analysis. APMIS 108:676–684
Flores-Diaz M, Alape-Giron A, Clark G, Catimel B, Hirabayashi Y, Nice E, Gutiérrez JM, Titball R, Thelestam M (2005) A cellular deficiency of gangliosides causes hypersensitivity to Clostridium perfringens phospholipase C. J Biol Chem 280:26680–26689
Johansson A, Aspán A, Kaldhusdal M, Engstrom B (2010) Genetic diversity and prevalence of netB in Clostridium perfringens isolated from a broiler flock affected by mild necrotic enteritis. Vet Microbiol 144:87–92
Jousimies-Somer HR, Summanen P, Citron DM, Baron EJ, Wexler HM, Finegold SM (2002) Wadsworth-KTL anaerobic bacteriology manual, 6th edn. Star publishing, California
Keyburn AL, Boyce JD, Vaz P, Bannam TL, Ford ME, Parker D, Di Rubbo A, Rood JI, Moore R (2008) NetB, a new toxin that is associated with avian necrotic enteritis caused by Clostridium perfringens. PLoS Pathog 4:1–11
Li J, Sayeed S, Robertson S, Chen J, McClane BA (2011) Sialidases affect host cell adherence and epsilon toxin-induced cytotoxicity of Clostridium perfringens type D strain CN3718. PLoS Pathog 7:e10024–e10029
Llanco LA, Nakano V, Ferreira AJP, Avila-Campos MJ (2012) Toxinotyping and antimicrobial susceptibility of Clostridium perfringens isolated from broiler chickens with necrotic enteritis. Int J Microbiol Res 4:290–294
Martin TG, Smyth JA (2009) Prevalence of netB among some clinical isolates of Clostridium perfringens from animals in the united states. Vet Microbiol 136:202–205
McGuckin MA, Linden SK, Sutton P, Florin TH (2011) Mucin dynamics and enteric pathogens. Nat Rev Microbiol 9:265–278
Moncla BJ, Braham P, Hillier SL (1990) Sialidase (neuraminidase) activity among gram-negative anaerobic and capnophilic bacteria. J Clin Microbiol 28:422–425
Muller HE, Werner H (1979) Die neuraminidase als pathogenetischer factor beieinem durch Bacteroides fragilis bedingten abszess. Z Med Mikrobiol Immunol 57:8–106
Nakano V, Piazza RMF, Avila-Campos MJ (2006) A rapid assay of the sialidase activity in species of the Bacteroides fragilis group by using peanut lectin hemagglutination. Anaerobe 12:238–241
Petit LM, Popoff GM (1999) Clostridium perfringens: toxinotype and genotype. Trends Microbiol 7:104–110
Sakurai J, Nagahama M, Oda M (2004) Clostridium perfringens alpha-toxin: characterization and mode of action. J Bio Chem 136:569–574
Salyers AA (1979) Energy sources of major intestinal fermentative anaerobes. Am J Clin Nutr 32:158–163
Sambrook J.K., Fritsch E., and Maniatis T. 1989. Molecular cloning. A laboratory manual. Book, Second edn., New York, p 1–3
Sawires Y, Songer J (2006) Clostridium perfringens: insight into virulence evolution and population structure. Anaerobe 12:23–43
Schauer R (1982) Chemistry, metabolism, and biological functions of sialic acids. Adv Carbohydr Chem Biochem 40:131–234
Sheu SY, Tseng HJ, Huang SP, Chien CH (2002) Cloning, expression, and deletion analysis of large nanH of Costridium perfringens ATCC 10543. En Microb Techss 31:794–803
Slavic D, Boerlin P, Fabri M, Klotins KC, Zoethout JK, Weir PE, Bateman D (2011) Antimicrobial susceptibility of Clostridium perfringens isolates of bovine, chicken, porcine, and turkey origin from Ontario. Can J Vet Res 75:89–97
Songer J (1996) Clostridial enteric diseases of domestic animals. Clin Microbiol Rev 9:216–234
Stackebrandt E, Kramer I, Swiderski J, Hippe H (1999) Phylogenetic basis for a taxonomic dissection of the genus Clostridium. FEMS Immunol Med Microbiol 24:253–258
Van Immerseel F, De Buck J, Pasmans F, Huyghebaert G, Haesebrouck F, Ducatelle R (2004) Clostridium perfringens in poultry: an emerging threat for animal and public health. Avian Pathol 33:537–549
Vimr ER, Kalivoda KA, Deszo EL, Steenbergen SM (2004) Diversity of microbial sialic acid metabolism. Microbiol Mol Biol Rev 68:132–153
Walters D, Stirewalt V, Melville S (1999) Cloning, sequence, and transcriptional regulation of the operon encoding a putative N-acetylmannosamine-6-phosphate epimerase (nanE) and sialic acid lyase (nanA) in Clostridium perfringens. J Bacteriol 181:4526–4532
Williams RB (2005) Intercurrent coccidiosis and necrotic enteritis of chickens: rational, integrated disease management by maintenance of gut integrity. Avian Pathol 34:159–180
Acknowledgments
The authors thank Marcia Harumi Fukugaiti for her technical support. During the course of this work VN was supported by FAPESP fellowship (2009/03792-0). This study was supported by Grant from Conselho Nacional Desenvolvimento Científico e Tecnológico (CNPq Proc. No. 143387/2008-1) and FAPESP (10/52417-4 and 13/17739-9).
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The authors declare no conflict of interest.
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Luis A. Llanco and Viviane Nakano have contributed equally to the work.
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Llanco, L.A., Nakano, V. & Avila-Campos, M.J. Sialidase Production and Genetic Diversity in Clostridium perfringens Type A Isolated from Chicken with Necrotic Enteritis in Brazil. Curr Microbiol 70, 330–337 (2015). https://doi.org/10.1007/s00284-014-0722-5
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DOI: https://doi.org/10.1007/s00284-014-0722-5