Characterization of coagulase-negative staphylococcus species from cows’ milk and environment based on bap, icaA, and mecA genes and phenotypic susceptibility to antimicrobials and teat dips

Piessens, Veerle; Vliegher, Sarne De; Verbist, Bert; Braem, Gorik; Nuffel, Annelies Van; Vuyst, Luc De; Heyndrickx, Marc; Coillie, Els Van

doi:10.3168/jds.2012-5400

Cited by 50 publications

(56 citation statements)

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“…Quarters from heifers and cows in peak lactation (61-120 DIM) were more likely to be infected with the more relevant species despite applying postmilking teat disinfection and using dry cow antibiotics in all 3 herds. This might support the assumption of CNS being able to develop resistance to antibiotics and teat disinfectants (Sampimon et al, 2011;Piessens et al, 2012). A high proportion of the variation in the likelihood of CNS IMI was observed at the quarter level, which is in agreement with the findings of another study conducted by our group (Passchyn et al, 2014).…”

Section: Short Communicationsupporting

confidence: 92%

Short communication: Species group-specific predictors at the cow and quarter level for intramammary infection with coagulase-negative staphylococci in dairy cattle throughout lactation

Visscher

Piepers

Supré³

et al. 2015

Journal of Dairy Science

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Coagulase-negative staphylococci (CNS) are frequently isolated from quarters with subclinical mastitis, teat apices, and the cows' environment. Virulence, ecology, epidemiological behavior, and effect on udder health vary between different CNS species. Staphylococcus chromogenes, Staph. simulans, and Staph. xylosus are frequently present in milk and have a more substantial effect on quarter milk somatic cell count than other species. Therefore, these species are considered the "more relevant" CNS. As species-specific factors associated with CNS intramammary infection (IMI) have not yet been identified and susceptibility for IMI differs between cows and quarters, this study aimed to identify predictors for CNS IMI at the cow and quarter level (some of them changing over time) with a specific focus on the aforementioned more relevant CNS. Precise data were available from a longitudinal study (3,052 observations from 344 quarters from 86 dairy cows belonging to 3 commercial dairy herds). All CNS were molecularly identified to the species level, and multivariable, multilevel logistic regression models taking into account the longitudinal nature of the data, were fit to study the likelihood of infection. Staphylococcus chromogenes, Staph. xylosus, and Staph. cohnii were the most frequently isolated species from CNS IMI in older cows, whereas Staph. chromogenes, Staph. xylosus, and Staph. simulans were the main species found in IMI in heifers. Quarters from heifers (as opposed to multiparous cows), from heifers and multiparous cows in third or fourth month in lactation (as opposed to early lactation, <60 d in milk), and with an increasing quarter milk SCC were more likely to be infected with the more relevant CNS species. Quarter milk SCC was identified as the sole statistically significant predictor for IMI with other CNS species, although the size of the effect was lower [odds ratio of 1.6 (1.4-1.9) vs. 2.1 (1.8-2.5)] than the effect for IMI with the more relevant CNS. As a strong herd effect was present, studying herd-level predictors is warranted.

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Section: Short Communicationsupporting

confidence: 92%

Short communication: Species group-specific predictors at the cow and quarter level for intramammary infection with coagulase-negative staphylococci in dairy cattle throughout lactation

Visscher

Piepers

Supré³

et al. 2015

Journal of Dairy Science

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“…Among CNS isolates, 20.1% were found to be resistant to 2 or more antibiotics, as reported by Rajala-Schultz et al (2004), and MDR frequencies of 7.9 and 10.6% were described by Rajala-Schultz et al (2004) and Sampimon et al (2011), respectively. With the exception of the resistance to penicillin, lower rates of resistance in CNS were reported by Botrel et al (2010), Persson et al (2011), andPiessens et al (2012). Few studies in Brazil have analyzed the antimicrobial susceptibility profile of staphylococci isolated from bovine milk using MIC, and the majority of them used the disk diffusion method.…”

Section: Discussionmentioning

confidence: 89%

“…However, we found higher MIC range values for CNS. Piessens et al (2012), who also used the Etest, reported enrofloxacin MIC 50 and MIC 90 of 0.125 and 0.25 µg/mL for CNS, respectively. In our study, a greater MIC 90 (0.5 µg/mL) was found.…”

Section: Discussionmentioning

confidence: 97%

“…Some species seem more adapted to the mammary gland, whereas others are more prevalent in the environment and rarely cause intramammary infection. It has been shown that antimicrobial resistance among the different species of CNS varies and was more frequently reported in species with environmental traits (Piessens et al, 2012). Unlike the cows infected with S. aureus, cows infected by CNS are less likely to be discarded as a mastitis control measure.…”

Section: Discussionmentioning

confidence: 99%

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Presence of mecA-positive multidrug-resistant Staphylococcus epidermidis in bovine milk samples in Brazil

Santos

Mendonça

Reis

et al. 2016

Journal of Dairy Science

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Bacteria of the genus Staphylococcus are one of the major pathogens causing bovine mastitis. In recent decades, resistance of this genus to oxacillin (methicillin) has been a matter of concern due to the possibility of reducing the effectiveness of mastitis treatments and the transfer of resistance determinants to other bacteria. Oxacillin resistance was studied in 170 staphylococci from bovine milk samples, including 79 Staphylococcus aureus and 91 coagulase-negative staphylococci (CNS). The susceptibility profile of 10 antimicrobial agents used in veterinary practice was determined by the Etest method. In addition to the Etest, the phenotypic characterization of oxacillin resistance was tested using the cefoxitin disk diffusion test. All isolates were screened by PCR to detect the mecA gene in 2 different regions of the gene. The isolates with an oxacillin minimum inhibitory concentration ≥0.5 µg/mL or resistant to cefoxitin were identified by sequencing a 536-bp fragment of the 16S rRNA gene. This group of isolates was also evaluated for the presence of blaZ and mecC genes. Molecular analysis of the mecA gene was carried out by typing of the staphylococcal cassette chromosome mec (SCCmec). The relatedness of the mecA-positive isolates was evaluated by macrorestriction of chromosomal DNA followed by pulsed-field gel electrophoresis. With the exception of penicillin and oxacillin, 86% of the isolates showed susceptibility to cephalothin, gentamicin, erythromycin, sulfonamide, trimethoprim-sulfamethoxazole, and tetracycline. All S. aureus isolates were susceptible to oxacillin, whereas 47% (n=43) of the CNS isolates were resistant. The CNS isolates showed a higher resistance to cephalothin, erythromycin, tetracycline, and gentamicin in comparison with S. aureus. The mecA gene was only detected in 10 CNS isolates, identified as Staphylococcus epidermidis, and classified into 3 pulsotypes (A, B, and C) and 4 subtypes (A1, B1, B2, and B3). Among the isolates with an oxacillin resistance phenotype, 12 were positive for the blaZ gene, and 9 of them were mecA-positive. Two of the oxacillin-resistant isolates amplified the mecA homolog gene of Staphylococcus sciuri and none amplified mecC. Three SCCmec types, I, IV, and V, were found. Our results suggest that Staphylococcus epidermidis can be a reservoir for mecA for other Staphylococcus species. Studies investigating the molecular and phenotypic profile of antimicrobial resistance in staphylococcal species should be performed for controlling the spread of resistance and the selection of appropriate therapeutic measures.

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“…We argue that a better methodology for biofilm detection is to screen strains for the icaD gene in addition to CRA or TA and MDCK methods to confirm the phenotypic expression. Piessens et al [34] considered that CNS species biofilm-forming strains might help in their survival in the environment and facilitate its transmission from the farm environment to the intramammary environment when the cow is lactating. The present results demonstrated the presence of intercellular adhesion genes (ica-AD) and consequent biofilm production in all CNS isolates.…”

Section: Resultsmentioning

confidence: 99%

Relevance of biofilm formation and virulence of different species of coagulase-negative staphylococci to public health

Osman

El-Razik²,

Marie³

et al. 2015

Eur J Clin Microbiol Infect Dis

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The ability of biofilm formation seems to play an essential role in the virulence of coagulase-negative staphylococci (CNS). The present work aimed to: (a) evaluate the biofilm-forming ability of different strains of CNS field isolates; (b) evaluate their virulence potential through the assessment of the Madin-Darby canine kidney (MDCK) cytotoxicity assay; (c) determine the frequency of biofilm-associated genes among these CNS isolates. Biofilm markers associated with biofilm formation and MDCK cells cytotoxicity were compared to find possible associations with pathogenicity. CNS isolates (n = 94) belonging to 11 different species were tested for slime production using the tube test (TA) and the Congo red agar plate test (CRA), while the presence of icaA and icaD genes were evaluated by d-PCR. Two points were addressed for the first time: (1) the specific relationship between slime phenotype and icaD gene expression; (2) the specific relationship between slime phenotype, icaAD genes, and MDCK cytotoxicity. The proportion of biofilm-positive/icaD-positive versus biofilm-positive/icaD-negative strains was 9:0 and 9:0 (81.8%) by the TA and CRA, which clearly indicates that icaD was a more reliable gene to be accounted for in the biofilm formation. MDCK recorded a higher proportion than that recorded by the CRA and TA results (MDCK-positive/icaD-positive versus MDCK-positive/icaD-negative 10:0, 90.9%). Evaluation of the ica operon, CRA plate test, TA, and MDCK can contribute to the high clinical impact in the management of antibiotic therapy, in infections associated with devices in veterinary medicine, the dairy industry, and food processing.

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Characterization of coagulase-negative staphylococcus species from cows’ milk and environment based on bap, icaA, and mecA genes and phenotypic susceptibility to antimicrobials and teat dips

Cited by 50 publications

References 40 publications

Short communication: Species group-specific predictors at the cow and quarter level for intramammary infection with coagulase-negative staphylococci in dairy cattle throughout lactation

Short communication: Species group-specific predictors at the cow and quarter level for intramammary infection with coagulase-negative staphylococci in dairy cattle throughout lactation

Presence of mecA-positive multidrug-resistant Staphylococcus epidermidis in bovine milk samples in Brazil

Relevance of biofilm formation and virulence of different species of coagulase-negative staphylococci to public health

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