Performance of API Staph ID 32 and Staph-Zym for identification of coagulase-negative staphylococci isolated from bovine milk samples

Sampimon, O.C.; Zadoks, Ruth N.; Vliegher, Sarne De; Supré, K.; Haesebrouck, Freddy; Barkema, Herman W.; Sol, J.; Lam, TH

doi:10.1016/j.vetmic.2008.11.004

Cited by 86 publications

(83 citation statements)

References 26 publications

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“…Some of this error could be due to storage error or loss of the initial organism during the recovery and subculturing of the isolates. Some of the error is also likely due to misclassification reported to occur with phenotypic species identification, as such methods can lead to a higher number of unidentified or misidentified staphylococcal isolates (27). When using phenotypic methods, S. chromogenes can be misidentified as S. hyicus (28), as was seen with 8/18 (44%) of the misclassified isolates in this study.…”

Section: Figmentioning

confidence: 58%

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

et al. 2017

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Staphylococcus hyicus and Staphylococcus agnetis are two coagulasevariable staphylococcal species that can be isolated from bovine milk and are difficult to differentiate. The objectives of this study were to characterize isolates of bovine milk origin from a collection that had previously been characterized as coagulase-positive S. hyicus based on phenotypic species identification methods and to develop a PCR-based method for differentiating S. hyicus, S. agnetis, and Staphylococcus aureus. Isolates (n ϭ 62) were selected from a previous study in which milk samples were collected from cows on 15 dairy herds. Isolates were coagulase tested and identified to the species level using housekeeping gene sequencing. A multiplex PCR to differentiate S. hyicus, S. agnetis, and S. aureus was developed. Pulsed-field gel electrophoresis was conducted to strain type the isolates. Based on gene sequencing, 44/62 of the isolates were determined to be either S. agnetis (n ϭ 43) or S. hyicus (n ϭ 1). Overall, 88% (37/42) of coagulase-positive S. agnetis isolates were found to be coagulase positive at 4 h. The herd-level prevalence of coagulasepositive S. agnetis ranged from 0 to 2.17%. Strain typing identified 23 different strains. Six strains were identified more than once and from multiple cows within the herd. Three strains were isolated from cows at more than one time point, with 41 to 264 days between samplings. These data suggest that S. agnetis is likely more prevalent on dairy farms than S. hyicus. Also, some S. agnetis isolates in this study appeared to be contagious and associated with persistent infections.KEYWORDS Staphylococcus, cattle, milk S taphylococci are the most common bacteria isolated from the cow's mammary gland and are frequently associated with subclinical or mild clinical mastitis (1-3). Staphylococci can be differentiated into groups using the coagulase test. Hemolytic coagulase-positive isolates are often presumptively identified as Staphylococcus aureus, a so-called "major" contagious mastitis pathogen, and coagulase-negative staphylococci are frequently grouped together as "minor" pathogens (4).Currently, there are two coagulase-variable staphylococcal species that have been associated with bovine mastitis, Staphylococcus hyicus and Staphylococcus agnetis. Staphylococcus hyicus was the first staphylococcal species to be described as coagulase variable, with coagulase production being found in approximately 24 to 56% of strains (5). The coagulation reaction of S. hyicus was described as often having weak activity,

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Section: Figmentioning

confidence: 58%

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

et al. 2017

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“…Nevertheless, Sampimona et al (2009) indicated that conventional methods such as the API Staph ID 32 and the Staph-Zym had a high number of unidentified or misidentified CNS strains. Both tests were inadequate to identify CNS isolates from mastitis milk samples.…”

Section: Ajmmentioning

confidence: 99%

Phenotypical and Mass Spectral Assessment Methods for Identification of Some Contagious Mastitis Pathogens

Behiry¹,

Zahran²,

Marzouk³

et al. 2014

American Journal of Microbiology

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Mastitis is one of the most economic disease affecting dairy cows worldwide. Identification of mastitis pathogens still depends principally on culture and phenotypical method, which is a difficult and timeconsuming. Newly, microbiologists have focused their attention on the use of Mass Spectrometry (MS) for microbial identification, especially Matrix Assisted Laser Desorption Ionization Time-Of-Flight (MALDI-TOF). Therefore, this study was designated to evaluate the ability of MALDI-TOF to identify some contagious mastitis pathogens comparing with phenotypical methods such as API panels and VITEK 2 system. A total of one hundred twenty of Staphylococcus aureus (S. aureus), Coagulase Negative Staphylococci (CNS) and Streptococcus agalactiae (Strept. agalactiae) strains isolated from milk of cows affected by clinical and subclinical mastitis were used in the study. According to the results, ~95% of S. aureus, 100% of CNS and Strept. agalactiae were correctly identified by MALDI TOF MS. All S. aureus isolates were then confirmed by a nuc-based PCR technique. While ~92% of S. aureus, 87% of Strept. agalactiae and 76% of CNS were identified by VITEK 2 system. Moreover, ~89% of S. aureus, 80% of Strept. agalactiae and 72% of CNS were identified by API system. In brief, the results demonstrated that MALDI-TOF is a fast and truthful technique which has the capability to replace conventional identification of several bacterial strains usually isolated in clinical laboratories of microbiology. Therefore, it is recomended that MALDI-TOF MS technology can be regularly used in veterinary laboratories for identification of different species of bacteria, particularly when failure of phenotypic methods forces clinical microbiologists.

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“…Apiweb software (bioMerieux) was used to determine the probability of species identification. Identification with a probability ≥ 90% was considered acceptable (Taponen et al 2006, Sampimon et al 2009b. Isolates for which the reliability of identification was <90% were then subjected to analysis with the VITEK GPI card system (VITEK 2 instrument, version 4.01, bioMérieux).…”

Section: Bacterial Identificationmentioning

confidence: 99%

Phenotypic and genotypic antimicrobial resistance of staphylococci from bovine milk

Kot¹,

Piechota²,

Wolska³

et al. 2012

Polish Journal of Veterinary Sciences

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The aim of this study was to examine phenotypic and genotypic antimicrobial resistance of staphylococci from milk samples from cows with subclinical and clinical mastitis and from cows without mastitis symptoms to methicillin, tetracyclines, macrolides and lincosamides (ML). Of 207 strains, including 34 S. aureus and 173 coagulase-negative staphylococci (CNS), 11 (6.4%) CNS strains were phenotypically resistant to methicillin. The mecA gene was detected by PCR only in two S. xylosus strains and one strain of S. epidermidis and S. simulans. No methicillin-resistant S. aureus strains were observed. In methicillin-resistant strains with mecA, gene resistance to other investigated antibiotics was not observed. Phenotypic resistance to tetracycline was detected in 11.0% of CNS strains and 47.4% of them carried the tetK gene. Of 173 CNS strains studied, 27 (15.6%) were resistant to at least one ML antibiotic. The resistance gene ermC was detected in 55.5% of the 27 ML-resistant strains. The ermA and ermB genes were detected in 14.8% and 11.1% of ML-resistant CNS strains, respectively. Antimicrobial resistance to methicillin, tetracyclines and macrolides was detected more frequently in staphylococcal strains from clinical mastitis compared to animals with subclinical symptoms and without mastitis, while the resistance to lincosamides showed a similar frequency in all groups of cows. In conclusion, CNS species from bovine milk differ in phenotypic and genotypic antimicrobial resistance profiles, and the use of PCR technique alone for the detection of methicillin, macrolide, lincosamide and tetyracycline resistance in CNS from cattle is not reliable.

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Performance of API Staph ID 32 and Staph-Zym for identification of coagulase-negative staphylococci isolated from bovine milk samples

Cited by 86 publications

References 26 publications

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

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

Phenotypical and Mass Spectral Assessment Methods for Identification of Some Contagious Mastitis Pathogens

Phenotypic and genotypic antimicrobial resistance of staphylococci from bovine milk

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