Genetically divergent methicillin-resistant Staphylococcus aureus and sec-dependent mastitis of dairy goats in Taiwan

BMC Veterinary Research

Yaochi Su²  Yanhaui Lee²  Changyou Yu²  Chishih Chu¹ 

[1] Department of Microbiology, Immunology, and Biopharmaceuticals, National Chiayi University, No. 300, University Road, Chiayi 60004, Taiwan;Department of Veterinary Medicine, National Chiayi University, No. 580, Sing Ming Road, Chiayi 60004, Taiwan

关键词: Goats;    Staphylococcal enterotoxin;    MDR;    Drug-resistance;    Mastitis;    Staphylococcus aureus;    MRSA;   

Others  :  1119848
DOI  :  10.1186/1746-6148-8-39

 received in 2011-06-24, accepted in 2012-03-29,  发布年份 2012

【 摘 要 】

Background

Widespread in the environment, Staphylococcus spp. infect animals and humans as normal flora or pathogens. By extending our recent report of multi-drug resistant (MDR) S. aureus in dairy goats, this study investigated the staphylococcal infection and characterized the MDR-S. aureus and methicillin-resistant S. aureus (MRSA) isolates collected from goats in 2008 to elucidate the appearance of MRSA in goats and the mastitis associated staphylococcus enterotoxin (SE) types. A total of 555 samples were collected from six goat parts and three environmental sources among four dairy goat farms in southern Taiwan. Coagulase-positive and negative Staphylococcus spp. (CPS and CNS, respectively) were also identified. Furthermore, predominant SE genes of nine enterotoxin genes sea through sej along with antimicrobial resistance and genetic variations were determined.

Results

In total, 137 staphylococcal strains were identified and found predominantly in milk, and in the vagina, anus, and nasal cavity. The most prevalent species was S. lentus, followed by S. aureus, S. epidermidis, and S. xylosus. Enterotoxin genes were not identified in any CNS isolates, however sec and see were identified only in S. aureus associated with mastitis in goat. In compared to the isolates from 2006 to 2007, 27 S. aureus isolates from 2008 were found to be more resistant to ampicillin, cephalothin, oxacillin, oxytetracycline, penicillin G, and tetracycline. Eleven MRSA isolates were identified and belonged to SCCmec type III (nine isolates) as the major type and SCCmec type II (two isolates). These MRSA isolates revealed pulse-field gel electrophoresis (PFGE) pattern A (five isolates), C (one isolate), and D (one isolate) of human isolates. The other two isolates without pulsotypes belonged to ST59.

Conclusion

The prevalence and infection sites of CNS differed from those of CPS. Genetic analyses indicated that genetic divergence, possible zoonotic transfer of MRSA, and the involvement of sec as important virulence factors for of S. aureus that lead to mastitis in goats.

【 授权许可】

   
2012 Chu et al; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

【 参考文献 】

• [1]Resch M, Nagel V, Hertel C: Antibiotic resistance of coagulase-negative staphylococci associated with food and used in starter cultures. Int J Food Microbiol 2008, 127:99-104.
• [2]Quinn PJ, Markey BK, Carter ME, Donnelly WJ, Leonard FC, Maguire D: Staphylococcus species. In Veterinary Microbiology and Microbial Disease. 1st edition. Edited by Quinn PJ. USA: Blackwell Science Ltd; 2002:43-48.
• [3]Abigail AS, Whitt DD: Bacterial Pathogenesis: A Molecular Approach. Washington: ASM Press; 2001.
• [4]Taponen S, Pyorala S: Coagulase-negative staphylococci as cause of bovine mastitis-not so different from Staphylococcus aureus? Vet Microbiol 2009, 134:29-36.
• [5]Krimmer V, Merkert H, von Eiff C, Frosch M, Eulert J, Löhr JF, Hacker J, Ziebuhr W: Detection of Staphylococcus aureus and Staphylococcus epidermidi in clinical samples by 16S rRNA-directed in situ hybridization. J Clin Microbiol 1999, 37:2667-2673.
• [6]Løvseth A, Loncarevic S, Berdal KG: Modified multiplex PCR method for detection of pyrogenic exotoxin genes in staphylococcal isolates. J Clin Microbiol 2004, 42:3869-3872.
• [7]Valle J, Gomez-Lucia E, Piriz S, Goyache J, Orden JA, Vadillo S: Enterotoxin production by Staphylococci isolated from healthy goats. Appl Environ Microbiol 1990, 56:1323-1326.
• [8]Lim SK, Joo YS, Moon JS, Lee AR, Nam HM, Wee SH, Koh HB: Molecular typing of enterotoxigenic Staphylococcus aureus isolated from bovine mastitis in Korea. J Vet Med Sci 2004, 66:581-584.
• [9]da Silva ER, do Carmo LS, da Silva N: Detection of the enterotoxins A, B, and C genes in Staphylococcus aureus from goat and bovine mastitis in Brazilian dairy herds. Vet Microbiol 2005, 106:103-107.
• [10]Reynolds PE, Brown DF: Penicillin-binding proteins of [beta]- lactam- resistant strains of Staphylococcus aureus: effect of growth conditions. FEBS Lett 1985, 192:28-32.
• [11]Geha DJ, Uhl JR, Gustaferro CA, Persing DH: Multiplex PCR for identification of methicillin-resistant staphylococci in the clinical laboratory. J Clin Microbiol 1994, 32:1768-1772.
• [12]Katayama Y, Ito T, Hiramatsu K: A new class of genetic element, staphylococcus cassette chromosome mec, encodes methicillin resistance in Staphylococcus aureus. Antimicrob Agents Chemother 2000, 44:1549-1555.
• [13]Oliveira DC, de Lencastre H: Multiplex PCR strategy for rapid identification of structural types and variants of the mec element in methicillin-resistant Staphylococcus aureus. Antimicrob Agents Chemother 2002, 46:2155-2161.
• [14]Chongtrakool P, Ito T, Ma XX, Kondo Y, Trakulsomboon S, Tiensasitorn C, Jamklang M, Chavalit T, Song JH, Hiramatsu K: Staphylococcal cassette chromosome mec (SCCmec) typing of methicillin-resistant Staphylococcus aureus strains isolated in 11 Asian countries: a proposal for a new nomenclature for SCCmec elements. Antimicrob Agents Chemother 2006, 50:1001-1012.
• [15]Boyle-Vavra S, Yin S, Challapalli M, Daum RS: Transcriptional induction of the penicillin-binding protein 2 gene in Staphylococcus aureus by cell wall-active antibiotics oxacillin and vancomycin. Antimicrob Agents Chemother 2003, 47:1028-1036.
• [16]Oliveira DC, Milheirico C, de Lencastre H: Redefining a structural variant of staphylococcal cassette chromosome mec, SCCmec type VI. Antimicrob Agents Chemother 2006, 50:3457-3459.
• [17]van Duijkeren E, Box AT, Heck ME, Wannet WJ, Fluit AC: Methicillin-resistant Staphylococci isolated from animals. Vet Microbiol 2004, 103:91-97.
• [18]Barrett FF, McGehee RF Jr, Finland M: Methicillin-resistant Staphylococcus aureus at Boston City Hospital - bacteriologic and epidemiologic observations. N Engl J Med 1968, 279:441-448.
• [19]Lin YC: Analysis of molecular typing Staphylococcus aureus of isolated from bacterimia patients. In Master Thesis. Tzu Chi University, Graduate Institute of Medical Science; 2005.
• [20]Seguin JC, Walker RD, Caron JP, Kloos WE, George CG, Hollis RJ, Jones RN, Pfaller MA: Methicillin-resistant Staphylococcus aureus outbreak in a veterinary teaching hospital: potential human-to-animal transmission. J Clin Microbiol 1999, 37:1459-1463.
• [21]Chu C, Lai JM, Yu CY, Chang WS, Lin HH, Su YC: Rapid identification of staphylococcal infection in dairy goat raw milk by a multiplex PCR assay. Taiwan Vet J 2010, 36:182-190.
• [22]Huang YC, Su LH, Wu TL, Liu CE, Young TG, Chen PY, Hseuh PR, Lin TY: Molecular epidemiology of clinical isolates of methicillin-resistant Staphylococcus aureus in Taiwan. J Clin Microbiol 2004, 42:307-310.
• [23]White EC, Hinckley LS: Prevalence of mastitis pathogens in goat milk. Small Ruminant Res 1999, 33:117-121.
• [24]Tung CC: Application of PCR-RFLP on molecular epidemiological study of Staphylococcus aureus isolated from milk of dairy cows and goats. In Master thesis. National Chung Hsing University, Department of Veterinary Medicine; 2004.
• [25]Bradley AJ, Green MJ: A study of the incidence and significance of Gram-positive infections acquired during the dry period under UK field conditions. In Proceedings of the 40th Annual Meeting of the National Mastitis Council: 11-14 February 2001. Nevada: Reno; 2001:185-186.
• [26]Sargeant JM, Scott HM, Leslie KE, Ireland MJ, Bashiri A: Clinical mastitis in dairy cattle in Ontario: frequency of occurrence and bacteriological isolates. Can Vet J 1998, 39:33-38.
• [27]Wilson DJ, Gonzalez RN, Das HH: Bovine mastitis pathogens in New York and Pennsylvania: prevalence and effects on somatic cell count and milk production. J Dairy Sci 1997, 80:2592-2598.
• [28]Workineh S, Bayleyegn M, Mekonnen H, Potgieter LN: Prevalence and aetiology of mastitis in cows from two major Ethiopian dairies. Trop Anim Health Prod 2002, 34:19-25.
• [29]Lee JH: Methicillin (Oxacillin)-resistant Staphylococcus aureus strains isolated from major food animals and their potential transmission to humans. Appl Environ Microbiol 2003, 69:6489-6494.
• [30]Gianneechini R, Concha C, Rivero R, Delucci I, Moreno LJ: Occurrence of clinical and sub-clinical mastitis in dairy herds in the west littoral region in Uruguay. Acta Vet 2002, 43:221-230. BioMed Central Full Text
• [31]Lin HH: Epidemiology for rapid identification, and genotyping, and antibiotic susceptibility of Staphylococcus aureus isolated from raw milk in dairy goats. In Master thesis. National Chiayi University, Department of Veterinary Medicine; 2008.
• [32]Erskine RJ, Eberhart RJ, Hutchinson LJ, Spencer SB: Herd management and prevalence of mastitis in dairy herds with high and low somatic cell counts. J Am Vet Med Assoc 1987, 190:1411-1416.
• [33]Hutton CT, Fox LK, Hancock DD: Mastitis control practices: differences between herds with high and low milk somatic cell counts. J Dairy Sci 1990, 73:1135-1143.
• [34]Kérouanton A, Hennekinne JA, Letertre C, Chesneau O, Brisabois A, De Buyser ML: Characterization of Staphlococcus aureus strains associated with food poisoning outbreaks in France. Int J Food Microbiol 2007, 115(3):369-375.
• [35]Holecková B, Holoda E, Fotta M, Kalinácova V, Gondoĺ J, Grolmus J: Occurrence of enterotoxigenic Staphlococcus aureus in food. Ann Agri Environ Med 2002, 9(2):179-182.
• [36]Pourmand MR, Memariani M, Hoseini M, Yazdchi SB: High prevalence of SEA gene among clinical isolates of Staphlococcus aureus in Tehran. Acta Med Iran 2009, 47(5):357-361.
• [37]Mizumachi E, Kato F, Hisatsune J, Tsuruda K, Uehara Y, Seo H, Sugai M: Clonal distribution of enterotoxingenic Staphlococcus aureus on handles of handheld shopping baskets in supermarkets. J Appl Microbial 2011, 110(2):562-567.
• [38]Chen TR, Hsiao MH, Chiou CS, Tsen HY: Development and use of PCR primers for the investigation of C1, C2 and C3 enterotoxin types of Staphylococcus aureus strains isolated from food-borne outbreaks. Int J Food Microbiol 2001, 71:63-70.
• [39]Rosec JP, Guiraud JP, Dalet C, Richard N: Enterotoxin production by Staphylococci isolated from foods in France. Int J Food Microbiol 1997, 35:213-221.
• [40]Bautista L, Gaya P, Medina M, Nenez M: A quantitative study of enterotoxin production by sheep milk Staphylococci. Appl Environ Microbiol 1988, 54:564-569.
• [41]Orden JA, Goyache J, Hernandez J, Domenech A, Suarez G, Gomez LE: Detection of enterotoxins and TSST-1 secreted by Staphylococcus aureus isolated from ruminant mastitis, comparison of ELISA and immunoblot. J Appl Bacteriol 1992, 72:486-489.
• [42]da Silva ER, do Carmo LS, da Silva N: Detection of the enterotoxins A, B, and C genes in Staphylococcus aureus from goat and bovine mastitis in Brazilian dairy herds. Vet Microbiol 2005, 106:103-107.
• [43]Kuroishi T, Komine KL, Kai K, Itagaki M, Koboyashi J, Ohta M, Kamata SK, Kumagai K: Concentrations and specific antibodies to staphylococcal enterotoxin-C and toxic shock syndrome toxin-1 in bovine mammary gland secretions, and inflammatory response to the intramammary inoculation of these toxins. Journal of Vet Med Sci 2003, 65:899-906.
• [44]Tondo EC, Guimaraes MC, Henriques JA, Ayub MA: Assessing and analyzing contamination of a dairy products processing plant by Staphylococcus aureus using antibiotic resistance and PFGE. Can J Microbiol 2000, 46:1108-1114.
• [45]White DG, Harman RJ, Matos JES, Langlois BE: Isolation and identification of coagulase negative staphylococcus species from bovine body sites and streak canals of nulliparous heifers. J Dairy Sci 1989, 72:1886-1892.
• [46]Wang SJ, Chow LW, Chen JH, Liang I: Distribution and identification of enterotoxigenic and non-enterotoxigenic Staphylococci isolated from goats and cattle. J Chin Soc Anim Sci 2002, 31:79-86.
• [47]Zhuang LP: Survey of the enterotoxigenic types for staphylococcal isolates obtained from goats, food and clinical samples. In Master thesis. National Chung Hsing University, Department of Food Science and Biotechnology; 1993.
• [48]Vautor E, Abadie G, Guibert JM, Huard C, Pépin M: Genotyping of Staphylococcus aureus isolated from various sites on farms with dairy sheep using pulsed-field gel electrophoresis. Vet Microbiol 2003, 96:69-79.
• [49]Juang ZW: Study of antibiotic resistance and epidemiological typing of Staphylococcus aureus isolated from bovine mastitis in Taiwan. In Master Thesis. National Chung Hsing University, Department of Veterinary Medicine; 2005.
• [50]Zadoks R, van Leeuwen W, Barkema H, Sampimon O, Verbrugh H, Schukken YH, van Belkum A: Application of pulsed-field gel electrophoresis and binary typing as tools in veterinary clinical microbiology and molecular epidemiologic analysis of bovine and human Staphylococcus aureus isolates. J Clin Microbiol 2000, 38:1931-1939.
• [51]Lange C, Cardoso M, Senczek D, Schwarz S: Molecular subtyping of Staphylococcus aureus isolates from cases of bovine mastitis in Brazil. Vet Microbiol 1999, 67:127-141.
• [52]Ito T, Katayama Y, Asada K, Mori N, Tsutsumimoto K, Tiensasitorn C, Hiramatsu K: Structural comparison of three types of staphylococcal cassette chromosome mec integrated in the chromosome in methicillin-resistant Staphylococcus aureus. Antimicrob Agents Chemother 2001, 45:1323-1336.
• [53]Davis SL, Rybak MJ, Amjad M, Kaatz GW, McKinnon PS: Characteristics of patients with healthcare-associated infection due to SCCmec type IV methicillin-resistant Staphylococcus aureus. Infect Control Hosp Epidemiol 2006, 27:1025-1031.
• [54]Cruz FD, Davies J: Horizontal gene transfer and the origin of species: lessons from bacteria. Trends Microbiol 2000, 8:128-133.
• [55]Lee JH: Methicillin (Oxacillin)-resistant Staphylococcus aureus strains isolated from major food animals and their potential transmission to humans. Appl Environ Microbiol 2003, 69:6489-6494.
• [56]Hsieh JM, Chen RS, Tsai TY, Pan TM, Chou CC: Phylogenetic analysis of livestock oxacillin-resistant Staphylococcus aureus. Vet Microbiol 2008, 126:234-242.
• [57]Chen RS: Analysis of staphlococcal enterotoxins from workers and livestock animals isolates. In Master Thesis. National Taiwan University, Graduate Institute of Veterinary Medicine; 2005.
• [58]Lowy FD: Staphylococcus aureus infections. N Engl J Med 1998, 339:520-532.
• [59]Le Loir Y, Baron F, Gautier M: Staphylococcus aureus and food poisoning. Genet Mol Res 2003, 31:63-76.
• [60]Joo YS, Fox LK, Davis WC, Bohach GA, Park YH: Staphylococcus aureus associated with mammary glands of cows: genotyping to distinguish different strain s among herds. Vet Microbiol 2001, 80:131-138.
• [61]Hsueh PR, Liu YC, Yang D, Yan JJ, Wu TL, Huang WK, Wu JJ, Ko WC, Leu HS, Yu CR, Luh KT: Multicenter surveillance of antimicrobial resistance of major bacterial pathogens in intensive care units in 2000 in Taiwan. Microb Drug Resist 2001, 7:373-382.
• [62]Waldner DN: Sample collection and handling. In Laboratory handbook on bovine mastiti. Edited by Hogan JS, Gonzalez RN, Harmon RJ, Nickerson SC, Oliver SP, Pankey JW, Smith KL. Wisconsin: National Mastitis Council; 1999:1-11.
• [63]Davidson TJ, Dohoo IR, Alan WD, Harry H, Kathy C: A cohort study of coagulase negative staphylococcal mastitis in selected dairy herds in Prince Edward Island. Can J Vet Res 1992, 56:275-280.
• [64]Myllys V, Ridell J, Bjorkroth J, Biese I, Pyoarla S: Persistence in bovine mastitis of Staphylococcus aureus clones as assessed by random amplified polymorphic DNA analysis, biotyping and ribotyping. Vet Microbiol 1997, 57:245-251.
• [65]Clinical and Laboratory Standards Institute: Performance standards for antimicrobial disc susceptibility tests; approved standard-9th ed. Wayne. 2006.
• [66]Song MD, Wachi M, Doi M, Ishino F, Matsuhashi M: Evolution of an inducible penicillin-target protein in methicillin-resistant Staphylococcus aureus by gene fusion. FEBS Lett 1987, 221:167-171.
• [67]Ito T, Katayama Y, Hiramatsu K: Cloning and nucleotide sequence determination of the entire mec DNA of pre-methicillin-resistant Staphylococcus aureus N315. Antimicrob Agents Chemother 1999, 43:1449-1458.
• [68]Betley MJ, Mekalanos JJ: Nucleotide sequence of the type A staphylococcal enterotoxin gene. J Bacteriol 1988, 170:34-41.
• [69]Jones CL, Khan SA: Nucleotide sequence of the enterotoxin B gene from Staphylococcus aureus. J Bacteriol 1986, 166:29-33.
• [70]Bohach GA, Schlievert PM: Nucleotide sequence of the staphylococcal enterotoxin C1 gene and relatedness to other pyrogenic toxins. Mol Gen Genet 1987, 209:15-20.
• [71]Bayles KW, Iandolo JJ: Genetic and molecular analyses of the gene encoding staphylococcal enterotoxin D. J Bacteriol 1989, 171:4799-4806.
• [72]Couch JL, Soltis MT, Betley MJ: Cloning and nucleotide sequence of the type E staphylococcal enterotoxin gene. J Bacteriol 1988, 170:2954-2960.
• [73]Munson SH, Tremaine MT, Betley MJ, Welch RA: Identification and characterization of staphylococcal enterotoxin types G and I from Staphylococcus aureus. Infect Immun 1998, 66:3337-3348.
• [74]Ren K, Bannan JD, Pancholi V, Cheung AL, Robbins JC, Fischetti VA, Zabriskie JB: Characterization and biological properties of a new staphylococcal exotoxin. J Exp Med 1994, 180:1675-1683.
• [75]Zhang S, Iandolo JJ, Stewart GC: The enterotoxin D plasmid of Staphylococcus aureus encodes a second enterotoxin determinant (sej). FEMS Microbiol Lett 1998, 168:227-233.
• [76]Nikolay S, Dmitriy V, Vladimir C, Avraham R: Simultaneous analysis of multiple staphylococcal enterotoxin genes by an oligonucleotide microarray assay. J Clin Microbiol 2004, 42:2134-2143.
• [77]Bannerman TL, Hancock GA, Tenover FC, Miller JM: Pulsed-field gel electrophoresis as a replacement for bacteriophage typing of Staphylococcus aureus. J Clin Microbiol 1995, 33:551-555.
• [78]Enright MC, Day NP, Davies CE, Peacock SJ, Spratt BG: Multilocus sequence typing for characterization of methicillin-resistant and methicillin-susceptible clones of Staphylococcus aureus. J Clin Microbiol 2000, 38:1008-1015.