Molecular Typing of Mastitis-Causing Staphylococcus aureus Isolated from Heifers and Cows

Castelani, Lívia; Santos, A. F. S. dos; Miranda, Mariana Dos Santos; Zafalon, L. F.; Pozzi, Cláudia Rodrigues; Arcaro, Juliana Rodrigues Pozzi

doi:10.3390/ijms14024326

Cited by 23 publications

(23 citation statements)

References 21 publications

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“…On the other hand, other PFGE clusters (except for cluster IX) were typically found only on one of the farms. Clonal spread of S. aureus mastitis isolates has been previously described at both the regional and farm level (Capurro et al 2010;Fessler et al 2010;Castelani et al 2013). Such clonal spread indicates that certain lineages of S. aureus mastitis strains have proven to be particularly successful.…”

Section: Discussionmentioning

confidence: 96%

Prevalence of mastitis pathogens in milk from clinically healthy cows

Cervinkova¹,

Vlkova²,

Borodacova³

et al. 2013

Vet. Med.

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A total of 669 individual cow milk samples originating from asymptomatic cows from 16 dairy farms were examined for the presence of microorganisms with the potential to cause mastitis. Coagulase-negative staphylococci clearly predominated (53.5% positive samples) followed by streptococci and enterococci (both occurring in 16.1% samples). Among streptococci, so-called mastitis streptococci (S. uberis, S. dysgalactiae and S. agalactiae) prevailed (11.7% positive samples). Enterobacteriaceae were found in 10.0% samples, most of which (6.6% samples) were positive for Escherichia coli. Yeasts (mainly Candida spp.) were found in 8.2% samples. One of the major mastitis pathogens, Staphylococcus aureus subsp. aureus, was isolated from 9.0% of samples. S. aureus isolates were further characterised in terms of their capability to form biofilm, antimicrobial susceptibility and clonality (PFGE). All S. aureus isolates were capable of biofilm formation and were generally susceptible to the majority of tested antibiotics. The exception was ampicillin, resistance to which was observed in 27.7% isolates. Therefore, the relatively frequent occurrence of S. aureus could be attributed to persistent intramammary infections due to biofilm formation rather than low efficacy of particular antibiotics. PFGE analysis revealed clonal spread of certain S. aureus isolates within and between farms indicating that certain lineages of S. aureus mastitis strains are particularly successful.

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Section: Discussionmentioning

confidence: 96%

Prevalence of mastitis pathogens in milk from clinically healthy cows

Cervinkova¹,

Vlkova²,

Borodacova³

et al. 2013

Vet. Med.

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“…aureus isolated from bovine milk is an important tool in epidemiological studies of mastitis and contributes to our understanding of the pathogen's dissemination (Castelani et al, 2013). As shown by a variety of strain-typing methods (Smith et al, 2005;Fournier et al, 2008;Graber et al, 2009), in most herds, one predominant strain of Staph.…”

Section: Introductionmentioning

confidence: 99%

Genomic characteristics of Staphylococcus aureus strains associated with high within-herd prevalence of intramammary infections in dairy cows

Cremonesi

Pozzi

Raschetti

et al. 2015

Journal of Dairy Science

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Staphylococcus aureus is one of the most important causes of mastitis in dairy cattle. Based on previous research, Staph. aureus genotypes with different pathogenic and contagious properties can cause intramammary infection (IMI) and coexist in the same herd. Our study aimed to compare Staph. aureus strains from herds that differed in IMI prevalence using different molecular approaches such as ribosomal spacer (RS)-PCR, multilocus sequence typing (MLST), spa typing, ribotyping, pulsed-field gel electrophoresis (PFGE), and multiplex PCR. For this purpose, 31 dairy herds with Staph. aureus IMI were selected, and 16 of these were chosen for a comparison study: the 8 high-prevalence (HP) herds had Staph. aureus IMI prevalence >28% and the 8 low-prevalence (LP) herds had an IMI prevalence <4%. A total of 650 isolates of Staph. aureus from mammary quarters of all positive cows were genotyped with RS-PCR, a technique based on amplification of a portion of the intergenic spacer 16S-23S rRNA, and a subset of 54 strains was also analyzed by multiplex PCR, ribotyping, PFGE, MLST, and spa typing. The RS-PCR analysis revealed 12 different profiles. Staphylococcus aureus strains isolated from 5 out of 8 HP herds showed a profile identical to the genotype B (GTB), described in previous studies as being strongly associated with high within-herd prevalence of Staph. aureus mastitis and the presence of the genes coding for enterotoxins sea, sed, and sej, a long x-region of spa gene, and 3 lukE fragments. Moreover, all strains isolated in the HP herds possessed genes coding for staphylococcal enterotoxins. In LP herds, a limited number of strains of 6 genotypes, different from those isolated in HP herds, were identified and GTB was not found. Within these genotypes, 4 strains were positive for the mecA gene. Preliminary results and comparison with other genotyping methods confirmed that genotyping by RS-PCR is an accurate, rapid, and inexpensive tool for future field studies on Staph. aureus mastitis strains and generates clinically relevant results.

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“…Using of this technique, most of bacterial species can be identified in hours, instead of days needed for classical culture methods. PCR has high sensitivity and specificity and can improve the level of detection (Boss et al, 2011;Castelani et al, 2013).…”

Section: Introductionmentioning

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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Molecular Typing of Mastitis-Causing Staphylococcus aureus Isolated from Heifers and Cows

Cited by 23 publications

References 21 publications

Prevalence of mastitis pathogens in milk from clinically healthy cows

Prevalence of mastitis pathogens in milk from clinically healthy cows

Genomic characteristics of Staphylococcus aureus strains associated with high within-herd prevalence of intramammary infections in dairy cows

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

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