Bulk tank milk from 58 dairy goat and sheep flocks located in southern Spain was examined to determine the prevalence and distribution of Staphylococci. A total of 45 isolates were obtained and characterized to determine the species, antimicrobial resistance profile, and genetic similitude by pulse-field gel electrophoresis (PFGE) using SmaI. Staphylococcus aureus isolates were confirmed by polymerase chain reaction (PCR) analysis of nuc, and resistance to methicillin was determined by PCR analysis of mecA. A total of 10 different staphylococcal species were identified, 22.2% and 77.8% of which were coagulase positive and negative, respectively. Twenty-two (48.89%) isolates were resistant to at least one antimicrobial agent. Higher antimicrobial resistance values were obtained against tetracycline (28.9%) and penicillin (22.2%). Two isolates (S. aureus and Staphylococcus lentus) were resistant to cefoxitin; however, none of the 45 isolates harbored mecA. Thirty pulsotypes were detected by PFGE. Interestingly, some isolates of S. aureus, S. lentus, Staphylococcus simulans, and Staphylococcus caprae showed high genetic similarity (>80%). These data suggest that genetically similar staphylococcal isolates circulate among goat and sheep dairy herds, and their different resistance patterns could be influenced by the management systems used.
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References
1.
AdkinsPRF, MiddletonJR, CalcuttMJ, StewartGC, FoxdLK. Species identification and strain typing of Staphylococcus agnetis and Staphylococcus hyicus isolates from bovine milk by use of a novel multiplex PCR assay and pulsed-field gel electrophoresis. J Clin Microbiol, 2017; 55:1778–1788.
2.
ArasZ, AydinI, KavK. Isolation of methicillin-resistant Staphylococcus aureus from caprine mastitis cases. Small Rumin Res, 2012; 102:68–73.
ClarkS, Mora-GarcíaMB. A 100 year review: Advances in goat milk research. J Dairy Sci, 2017; 100:10026–10044.
5.
Clinical and Laboratory Standards Institute. Performance Standards for Antimicrobial Disk and Dilution Susceptibility Test for Bacteria Isolated from Animals. Third edition. CLSI Document VET01S. Wayne, PA: CLSI, 2013.
6.
CusatoS, GameiroAH, CorassinCH, Sant'AnaAS, CruzAG, FariaJAF, de OliveiraCAF. Food safety systems in a small dairy factory: Implementation, major challenges, and assessment of systems' performances. Foodborne Pathog Dis, 2013; 10:6–12.
7.
Da SilvaER, SiqueiraAP, MartinsJCD, FerreiraWPB, Da SilvaN. Identification and in vitro antimicrobial susceptibility of Staphylococcus species isolated from goat mastitis in the Northeast of Brazil. Small Rumin Res, 2004; 55:45–49.
8.
De BlasI. WinEpi 2.0 statistic program. Working in Epidemiology. Zaragoza, Spain: Facultad de Veterinaria de la Universidad de Zaragoza. 2006.
9.
De VisscherA, SupréK, HaesebrouckF, ZadoksRN, PiessensV, Van CoillieE, PiepersS, De VliegherS. Further evidence for the existence of environmental and host-associated species of coagulase-negative staphylococci in dairy cattle. Vet Microbiol, 2014; 172:466–474.
10.
DjoudiF, BonuraC, TouatiA, AléoA, BenallaouaS, MamminaC. Staphylococcal cassette chromosome mec typing and mecA sequencing in methicillin-resistant staphylococci from Algeria: A highly diversified element with new mutations in mecA. J Med Microbiol, 2016; 65:1267–1273.
11.
FrançaCA, PeixotoRM, CavalcanteMB, MeloNF, OliveiraCJB, VeschiJLA, MotaRA, CostaMM. Antimicrobial resistance of Staphylococcus spp. from small ruminant mastitis in Brazil. Pesquisa Veterinaria Brasileira, 2012; 32:747–753.
12.
HunterPR. Reproducibility and indices of discriminatory power of microbial typing methods. J Clin Microbiol, 1990; 28:1903–1905.
13.
KatayamaY, ItoT, HiramatsuK. A new class of genetic element, Staphylococcus Cassette Chromosome mec, encodes Methicillin resistance in Staphylococcus aureus. Antimicrob Agents Chemoter, 2000; 44:1549–1555.
14.
KateeteDP, KimaniCN, KatabaziFA, OkengA, OkeeMS, NantezaA, JolobaML, NajjukaFC. Identification of Staphylococcus aureus: DNase and mannitol salt agar improve the efficiency of the tube coagulase test. Ann Clin Microbiol Antimicrob, 2010; 13:9–23.
15.
KunzF, CortiS, GiezendannerN, StephanR, WittenbrinkM, ZweifelC. Antimicrobial resistance of Staphylococcus aureus and coagulase negative staphylococci isolated from mastitis milk samples from sheep and goats. Schweiz Arch Tierheilkd, 2011; 153:63–69.
16.
LadSS, AparnathiKD, MehtaB, VelpulaS. Goat milk in human nutrition and health. A review. Int J Curr Microbiol App Sci, 2017; 6:1781–1792.
17.
MagiorakosAP, SrinivasanA, CareyRB, CarmeliY, FalagasME, GiskeCG, HarbarthS, HindlerJF, KahlmeterG, Olsson-LiljequistB, PatersonDL, RiceLB, StellingJ, StruelensMJ, VatopoulosA, WeberJT, MonnetDL. Multidrug-resistant, extensively drug-resistant and pandrug-resistant bacteria: An international expert proposal for interim standard definitions for acquired resistante. Clin Microbiol Infect, 2011; 18:268–281.
18.
MartinsKB, Faccioli-MartinsPY, RiboliDF, PereiraVC, FernandesS, OliveiraAA, DantasA, ZafalonLF, CunhaMLRS. Clonal profile, virulence and resistance of Staphylococcus aureus isolated from sheep milk. Braz J Microbiol, 2015; 46:535–43.
19.
MartinsKB, FaccioliPY, BonessoMF, FernandesS, OliveiraAA, DantasA, ZafalonLF, CunhaMLRS. Characteristics of resistance and virulence factors in different species of coagulase-negative staphylococci isolated from milk of healthy sheep and animals with subclinical mastitis. J Dairy Sci, 2016; 100:2184–2195.
20.
McdougalLK, StewardCD, KillgoreGE, ChaitramJM, McallisterSK, TenoverFC. Pulsed-field gel electrophoresis typing of oxacillin-resistant Staphylococcus aureus isolated from the United States: Establishing a national database. J Clin Microbiol, 2003; 41:5113–5120.
21.
MoroniP, PisoniG, AntoniniM, RuffoG, CarliS, VariscoG, BoettcherP. Subclinical mastitis and antimicrobial susceptibility of Staphylococcus caprae and Staphylococcus epidermidis isolated from two Italian goat herds. J Dairy Sci, 2005; 88:1694–1704.
22.
Olde RiekerinkRGM, BarkemaHW, SchollDT, PooleDE, KeltonDF. Management practice associated with the bulk-milk prevalence of Staphylococcus aureus in Canadian dairy farms. Prev Vet Med, 2010; 97:20–28.
23.
OlechnowiczJ, JaskowskiJM. Mastitis in small ruminants. Med Weter, 2014; 70:67–72.
24.
PengovA, CeruS. Antimicrobial drug susceptibility of Staphylococcus aureus strains isolated from bovine and ovine mammary glands. J Dairy Sci, 2003; 86:3157–3163.
25.
PatersonGK, HarrisonEM, HolmesMA. The emergence of mecC methicillin-resistant Staphylococcus aureus. Trends Microbiol, 2014; 22:42–7.
26.
PulinaG, MilánMJ, LavínMP, TheodoridisA, MorinE, CapoteJ, ThomasDL, FrancesconiAHD, CajaG. Invited review: Current production trends, farm structures, and economics of the dairy sheep and goat sectors. J Dairy Sci, 2018; 101:6715–6729.
27.
Regulation (EC) No 178/2002 of the European Parliament and of the council of 28 January 2002 laying down the general principles and requirements of food law, establishing the European Food Safety Authority and laying down procedures in matters of food safety. OJ L 031, 01/02/2002, p. 1–24. http://data.europa.eu/eli/reg/2002/178/oj Accessed May28, 2019.
28.
Regulation (EC) No 852/2004 of the European Parliament and of the Council of 29 April 2004 on the hygiene of foodstuffs. OJ L 139, 30/4/2004, p. 1–54. http://data.europa.eu/eli/reg/2004/852/oj Accessed May28, 2019.
29.
Regulation (EC) No 853/2004 of the European Parliament and of the Council of 29 April 2004 laying down specific hygiene rules for food of animal origin. OJ L 139, 30/4/2004, p. 55–205. http://data.europa.eu/eli/reg/2004/853/oj Accessed May28, 2019.
30.
Regulation (EC) No 854/2004 of the European Parliament and of the Council of 29 April 2004 laying down specific rules for the organisation of official controls on products of animal origin intended for human consumption. OJ L 139, 30/4/2004, p. 206–320. http://data.europa.eu/eli/reg/2004/854/oj Accessed May28, 2019.
31.
Regulation (EU) 2017/625 of the European Parliament and of the Council of 15 March 2017 on official controls and other official activities performed to ensure the application of food and feed law, rules on animal health and welfare, plant health and plant protection products, amending Regulations (EC) No 999/2001, (EC) No 396/2005, (EC) No 1069/2009, (EC) No 1107/2009, (EU) No 1151/2012, (EU) No 652/2014, (EU) 2016/429 and (EU) 2016/2031 of the European Parliament and of the Council, Council Regulations (EC) No 1/2005 and (EC) No 1099/2009 and Council Directives 98/58/EC, 1999/74/EC, 2007/43/EC, 2008/119/EC and 2008/120/EC, and repealing Regulations (EC) No 854/2004 and (EC) No 882/2004 of the European Parliament and of the Council, Council Directives 89/608/EEC, 89/662/EEC, 90/425/EEC, 91/496/EEC, 96/23/EC, 96/93/EC and 97/78/EC and Council Decision 92/438/EEC (Official Controls Regulation) Text with EEA relevance. OJ L 95, 7/4/2017, p. 1–142. http://data.europa.eu/eli/reg/2017/625/oj Accessed May28, 2019.
32.
SmibertRM, KriegNR. Methods for General and Molecular Bacteriology. Washington, DC: American Society for Microbiology, 1994.
33.
StevensS, PiepersS, SupréK, de VliegherS. Antimicrobial consumption on dairy herds and its association with antimicrobial inhibition zone diameters of non-aureus staphylococci and Staphylococcus aureus isolated from subclinical mastitis. J Dairy Sci, 2018; 101:3311–3322.
34.
TehKH, FlintS, PalmerJ, LindsayD, AndrewesP, BremerP. Thermo-resistant enzyme-producing bacteria isolated from the internal surfaces of raw milk tankers. Int Dairy J, 2011; 21:742–747.
35.
TenoverFC, ArbeitRD, GoeringRV, MickelsenPA, MurrayBE, PersingDH, SwaminathanB. Interpreting chromosomal DNA restriction patterns produced by pulsed-field gel electrophoresis: Criteria for bacterial strain typing. J Clin Microbiol, 1995; 33:2233–2239.
36.
VanderhaeghenW, PiepersS, LeroyF, Van CoillieE, HaesebrouckF, De VliegherS. Identification, typing, ecology and epidemiology of coagulase negative staphylococci associated with ruminants. Vet J, 2015; 203:44–51.
37.
VirdisS, ScaranoC, CossuF, SpanuV, SpanuC, De SantisEPL. 2010. Antibiotic resistance in Staphylococcus aureus and coagulase negative staphylococci isolated from goats with subclinical mastitis. Vet Med Int, 2010; 2010:517060.
38.
WieldersCL, FluitAC, BrisseS, VerhoefJ, SchmitzFJ. mecA gene is widely disseminated in Staphylococcus aureus population. J Clin Microbiol, 2002; 40:3970–3975.
39.
XuZ, ShiL, AlamMJ, LiL, YamasakiS. Integron-bearing methicillin-resistant coagulase-negative staphylococci in South China, 2001–2004. FEMS Microbiol Lett, 2008; 278:223–230.
40.
ZadoksRN, WattsJL. Species identification of coagulase-negative staphylococci: Genotyping is superior to phenotyping. Vet Microbiol, 2009; 134:20–28.
