scholarly journals Staphylococcus aureus and bovine mastitis: molecular typing of methicillinresistance and clinical description of infected quarters

2019 ◽  
Vol 70 (2) ◽  
pp. 1511
Author(s):  
Α. OREIBY ◽  
H. KHALIFA ◽  
A. EID ◽  
A. AHMED ◽  
T. SHIMAMOTO ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Methicillin Resistance ◽  
Animal Health ◽  
Bovine Mastitis ◽  
Clinical Signs ◽  
Subclinical Mastitis ◽  
Clinical Severity ◽  
Resistance Pattern ◽  
Gram Positive ◽  
Staphylococcus Spp

This study targeted Staphylococcus aureus (S. aureus)-mastitis of bovine as a possible source of livestock-associated methicillin-resistant Staphylococcus aureus (MRSA), to describe clinical signs of mastitis associated with MRSA and methicillin-sensitive Staphylococcus aureus (MSSA). The study area was the Gharbia and Kafrelsheikh governorates, in the central and northern regions of the Egyptian Delta. Clinical examination of animals was performed to detect clinical mastitis (CM) and clinically normal cases were tested by the California mastitis test (CMT) to identify subclinical mastitis (SCM). Accordingly, 38 mastitis cases (68 infected quarters) were detected. Milk samples were collected aseptically and were cultured on Baird Parker agar. Fifty nine Gram-positive cocci-shaped isolates were selected and preserved. In addition, 33 Staphylococcus spp. isolates originated from bovine mastitis at the same study area were obtained from Animal Health Research Institute (AHRI). The cocci-shaped Gram-positive bacteria and AHRI Staphylococcus spp. isolates were used for molecular identification of S. aureus and MRSA. Molecular screening had yielded 17 S. aureus isolates, from which five isolates (29.41%) were MRSA and 12 isolates (70.59%) were MSSA. The five MRSA isolates were mecA positive, but mecC negative. Multilocus sequence typing (MLST) of the five MRSA isolates indicated that all were sequence type 1 (ST1). S. aureus-associated cases showed different clinical forms of mastitis, including subclinical, acute, chronic, and gangrenous. However, subclinical mastitis was the only detected form associated with MRSA, which may represent a potential hidden risk for humans. Phenotypic antimicrobial resistance pattern of MRSA isolates showed resistance to all of the tested β-lactam antimicrobials, with marked orgaresistance to tetracycline and gentamycin. Based on our knowledge, this is the first report to identify MRSA ST1 in Egypt. Bovine mastitis could be a source for the dissemination of MRSA to humans and other animals. Additionally,while methicillin-resistance may have no effect on the clinical severity of mastitis, it does affect therapeutic success, particularly when β-lactam antimicrobials are used.

scholarly journals Clinical and molecular characterization of both methicillin-resistant andsensitive staphylococcus aureus mastitis

2019 ◽  
Vol 70 (3) ◽  
pp. 1743
Author(s):  
A. OREIB ◽  
H. KHALIFA ◽  
A. EID ◽  
A. AHMED ◽  
T. SHIMAMOTO ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Methicillin Resistance ◽  
Bovine Mastitis ◽  
Clinical Signs ◽  
Subclinical Mastitis ◽  
Therapeutic Success ◽  
Methicillin Resistant ◽  
Marked Resistance

This study targeted bovine mastitis as a possible source of livestock-associated methicillin-resistant Staphylococcus aureus (MRSA), to identify clinical signs associated with MRSA- and non-MRSA-associated mastitis. Thirty-eight mastitis cases (68 infected quarters) were investigated. Gram-positive cocci-shaped isolates were selected based on Baird Parker agar growth as well as Gram-stained bacterial smears. Molecular screening for Staphylococcus aureus (S. aureus) yielded 17 isolates, of which five (29.41%) were methicillin resistant. The five isolates were mecA positive, but mecC negative. Multilocus sequence typing (MLST) indicated that sequence type 1 (ST1) was the identified type of all isolates of MRSA. S. aureus-associated cases showed different clinical forms of mastitis, including subclinical, acute, chronic, and gangrenous. Additionally, subclinical mastitis was the only detected condition associated with MRSA, which may represent a potential hidden risk for humans. Phenotypically, isolates of MRSA showed resistance to all of the tested β-lactam antimicrobials, with marked resistance to tetracycline and gentamycin. Based on our knowledge, this is the first report to identify MRSA ST1 in Egypt. Bovine mastitis could be a source for the dissemination of MRSA to humans and other animals. Additionally, while methicillin-resistance may have no effect on the clinical outcome of mastitis, it does affect therapeutic success, particularly when β-lactam antimicrobials are used.

scholarly journals Expression of icaA and icaD genes in biofilm formation in Staphylococcus aureus isolates from bovine subclinical mastitis

2021 ◽  
Vol 41 ◽  
Author(s):  
Viviane F. Marques ◽  
Huarrisson A. Santos ◽  
Thomas H. Santos ◽  
Dayanne A. Melo ◽  
Shana M.O. Coelho ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Biofilm Formation ◽  
Phenotypic Expression ◽  
Bovine Mastitis ◽  
Subclinical Mastitis ◽  
Transcriptional Analysis ◽  
Important Species ◽  
High Prevalence ◽  
Staphylococcus Spp

ABSTRACT: Staphylococcus spp. plays a significant role in the etiology of bovine mastitis. Staphylococcus aureus is considered the most important species due to the high prevalence and the difficulty of in vivo treatment that is related to the expression of virulence factors and biofilm formation. This study aimed to detect the phenotypic expression of the biofilm formation in 20 S. aureus isolated from bovine mastitis and to evaluate the expression and regulation of genes involved in its production. MALDI-TOF and phenogenotypic identification assays were performed to characterize the isolates. The phenotypic biofilm production and the presence of icaA and icaD and bap genes were evaluated. The Agr system was typified (agr I, agr II, agr III and agr IV) and its regulator (agr RNAIII) was detected. Furtherly, Real-time PCR (qPCR) was performed at chosen times to quantify the expression of icaA, icaD and hld genes in three selected isolates. All 20 strains were biofilm producers and most presented icaA and icaD genes. Only one isolate presented the bap gene. The agr gene type II showed a prevalence of 70%. Transcriptional analysis revealed increased expression of ica genes at eight hours of growth. These results confirm that polysaccharides production mediated by the icaADBC operon genes is an essential mechanism to the biofilm formation and contributes to the early stages of bacterial growth.

scholarly journals Virulence factors and antimicrobial resistance of Staphylococcus aureus isolated from bovine mastitis in Rio de Janeiro

2009 ◽  
Vol 29 (5) ◽  
pp. 369-374 ◽  
Author(s):  
Shana M.O. Coelho ◽  
Elina Reinoso ◽  
Ingrid A. Pereira ◽  
Lidiane C. Soares ◽  
Mirta Demo ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Virulence Factors ◽  
Protein A ◽  
Bovine Mastitis ◽  
Subclinical Mastitis ◽  
Resistance Pattern ◽  
Igg Binding ◽  
Amplicon Size ◽  
Resistance To Penicillin ◽  
Single Amplicon

The study was conducted to characterize pheno-genotypically the virulence factors and resistance pattern of Staphylococcus aureus isolates from milk samples of cows with subclinical mastitis. All hemolytic isolates presented beta-hemolysin, and 38% of the non-hemolytic isolates were able to express hemolysins in the presence of a beta-hemolytic strain. The amplification of the coa-gene displayed four different size polymorphisms with about 400 bp, 600 bp, 700 bp and 900 bp. The spaA gene that encodes the IgG-binding region of protein A revealed sizes of 700 bp and 900 bp. The amplification of region X from spaA yielded a single amplicon for each isolate with the prevalent amplicon size being of 180 bp. Amplification of sae gene yielded an amplicon size of 920 bp in 71% of the isolates. Antibiotic resistance pattern revealed that 42% S. aureus were susceptible to all antimicrobials tested. Seven different antibiotic patterns were observed. Our results indicated that 47% and 25% of S. aureus strains exhibited resistance to penicillin and oxacillin respectively. All oxacillin-resistant isolates were mecA-positive.

scholarly journals Genomic analysis of European bovine Staphylococcus aureus from clinical versus subclinical mastitis

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Jurriaan Hoekstra ◽  
Aldert L. Zomer ◽  
Victor P. M. G. Rutten ◽  
Lindert Benedictus ◽  
Arjan Stegeman ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Clinical Outcome ◽  
Virulence Genes ◽  
Mixed Model ◽  
Linear Mixed Model ◽  
Genome Wide Association Study ◽  
Bovine Mastitis ◽  
Subclinical Mastitis ◽  
Clinical Severity ◽  
A Genome

Abstract Intramammary infections (IMI) with Staphylococcus aureus are a common cause of bovine mastitis and can result in both clinical (CM) or subclinical mastitis (SCM). Although bacterial isolates of S. aureus differ in their virulence potential it is largely unclear which bacterial virulence factors are responsible for increased clinical severity. We performed a genome wide association study and used a generalized linear mixed model to investigate the correlation between gene carriage, lineage and clinical outcome of IMI in a collection of S. aureus isolates from cattle with CM (n = 125) and SCM (n = 151) from 11 European countries. An additional aim was to describe the genetic variation of bovine S. aureus in Europa. The dominant lineages in our collection were clonal complex (CC) 151 (81/276, 29.3%), CC97 (54/276, 19.6%), CC479 (32/276, 11.6%) and CC398 (19/276, 6.9%). Virulence and antimicrobial resistance (AMR) gene carriage was highly associated with CC. Among a selection of nine virulence and AMR genes, CC151, CC479 and CC133 carried more virulence genes than other CCs, and CC398 was associated with AMR gene carriage. Whereas CC151, CC97 were widespread in Europe, CC479, CC398 and CC8 were only found in specific countries. Compared to CC151, CC479 was associated with CM rather than SCM (OR 3.62; 95% CI 1.38–9.50) and the other CCs were not. Multiple genes were associated with CM, but due to the clustering within CC of carriage of these genes, it was not possible to differentiate between the effect of gene carriage and CC on clinical outcome of IMI. Nevertheless, this study demonstrates that characterization of S. aureus CC and virulence genes helps to predict the likelihood of the occurrence of CM following S. aureus IMI and highlights the potential benefit of diagnostics tools to identify S. aureus CC during bovine mastitis.

scholarly journals Expression of genes encoding resistance in Staphylococcus spp. isolated from bovine subclinical mastitis in Brazil

2020 ◽  
Vol 14 (07) ◽  
pp. 772-780
Author(s):  
Eveline Zuniga ◽  
Nilson Roberti Benites ◽  
Aline Santana da Hora ◽  
Priscila Luiza Mello ◽  
Marco Antonio Laes ◽  
...  
Keyword(s):  
Methicillin Resistance ◽  
Meca Gene ◽  
Bovine Mastitis ◽  
Subclinical Mastitis ◽  
Gene Encoding ◽  
Resistance Factors ◽  
Expression Of Genes ◽  
Genes Encoding ◽  
Staphylococcus Spp

Introduction: Staphylococci are the most important agents associated with bovine mastitis. This study aimed at characterizing resistance factors to antimicrobials in Staphylococcus spp. isolated from the milk of cows with subclinical mastitis. Methodology: In vitro resistance of 243 Staphylococcus spp. isolates to antimicrobials commonly used in clinical practice was evaluated. The detection and expression of genes encoding resistance mecA (gene encoding penicillin binding protein 2a) mecALGA251 (mecA homologue), blaZ (gene encoding penicillin resistance), femA and femB (genes encoding essential factors - A and B - for the expression of methicillin resistance) and aacA-aphD (gene encoding for a bifunctional enzyme that confers resistance to gentamicin) using PCR and RT-PCR was investigated. Results: One or more genes encoding resistance to different antimicrobials were detected in 184 Staphylococcus spp. samples. The femA and femB genes were the most frequent. Regarding the variables’ detection (N = number of strains) and expression (% of strains), the following results were obtained: blaZ (N = 40 – 82.5%), femA (N = 147 – 47.6%), aacAaphD (N = 30 – 43.3%), femB (N = 138 – 29.7%), mecA (N = 33 – 27.3%), mecALGA251 (N = 01 – 0.0%). There was a higher occurrence of phenotypic resistant strains for amoxicillin, ampicillin and penicillin in isolates positive for detection and/or expression of blaZ gene when compared with the other genes. Conclusions: The present study provides new information on genotypic traits of Staphylococcus isolates from bovine subclinical mastitis especially regarding the evaluation of expression of genes associated with antimicrobial resistance in Staphylococcus spp. using molecular tools.

scholarly journals The Prevalence and Antimicrobial Susceptibility Pattern of Gram-Positive Pathogens: Three-Year Study at a Private Tertiary Care Hospital in Mumbai, India

2021 ◽  
Author(s):  
Sweta Shah ◽  
Ritika Rampal ◽  
Pooja Thakkar ◽  
Sushima Poojary ◽  
Shweta Ladi
Keyword(s):  
Antimicrobial Susceptibility ◽  
Tertiary Care Hospital ◽  
Methicillin Resistance ◽  
Tertiary Care ◽  
Resistance Pattern ◽  
Steady Increase ◽  
Coagulase Negative Staphylococcus ◽  
Care Hospital ◽  
Gram Positive ◽  
Tract Infections

Abstract Introduction The growing resistance pattern of the gram-positive pathogens along with a steady increase in minimum inhibitory concentration of the currently available antibiotics have led to an increase in morbidity and mortality rates in India. This study aims to access the shifting antibiotic susceptibility paradigm of the gram-positive pathogens in various infections at a tertiary care center. Methods This is a 3-year retrospective observational study which was performed from January 2016 to December 2018 at a tertiary care hospital in Mumbai. All clinically significant gram-positive cocci isolated from a variety of clinical specimens were studied for their prevalence and antimicrobial susceptibility. Results Out of 4,428 gram-positive isolates, Staphylococcus aureus (35.3%) was the commonly encountered pathogen, followed by Enterococcus spp. (32.1%) and coagulase-negative Staphylococcus (CoNS) (25.7%). S. aureus was majorly isolated from skin and soft tissue infections (60.3%), followed by patients with respiratory tract infections (18.2%) and blood stream infections (13%). Among S. aureus, particularly methicillin-resistant S.aureus (MRSA), prevalence increased from 29.5% in 2016 to 35.1% in 2018, with an overall prevalence of 33.6%. All S. aureus isolates were 100% sensitive toward vancomycin, linezolid, tigecycline, and teicoplanin. However, the CoNS isolates showed a higher resistance rate with reduced susceptibility toward linezolid and teicoplanin. High prevalence of resistance was observed across gram-positive isolates with commonly used antibiotics such as ciprofloxacin, levofloxacin, and erythromycin. While the prevalence of linezolid-resistant enterococcus (LRE) was 3.6%, vancomycin (VRE) and teicoplanin resistance among the enterococcus species was as high as 7.7% and 7.5%, respectively. Conclusion Rising methicillin resistance among the Staphylococcal species (MRSA and MR-CoNS) along with reduced susceptibility toward currently available anti-MRSA agents is a matter of serious concern as it limits the therapeutic options for treating multidrug resistant (MDR) gram-positive infections.

scholarly journals Molecular characterization of virulence factors in Staphylococcus aureus isolated from bovine subclinical mastitis in central Ethiopia

2021 ◽  
Vol 71 (1) ◽  
Author(s):  
Desiye Tesfaye Tegegne ◽  
Gezahegne Mamo ◽  
Hika Waktole ◽  
Yohannes Equar Messele
Keyword(s):  
Staphylococcus Aureus ◽  
Dairy Cows ◽  
Virulence Factors ◽  
Virulence Genes ◽  
Animal Health ◽  
Subclinical Mastitis ◽  
Biochemical Tests ◽  
Milk Samples ◽  
Lactating Dairy Cows ◽  
Genomic Study

Abstract Purpose Staphylococcus aureus (S. aureus) is the most important pathogen involved in bovine mastitis in dairy production. S. aureus produces a spectrum of extracellular protein toxins and virulence factors which are thought to contribute to the pathogenicity of the organism. The aim of this work was to isolate and molecular characterize S. aureus associated with bovine subclinical mastitis in the central part of Ethiopia. Methods A total of 265 lactating dairy cows from various dairy farms in four different geographical locations were screened by the California mastitis test (CMT) for bovine subclinical mastitis. One hundred thirty CMT-positive milk samples were collected and transported to the laboratory. Different biochemical tests and polymerase chain reaction (PCR) were used for the identification of S. aureus isolates. Finally, PCR was performed for molecular detection of virulence genes. Results From a total of 265 lactating dairy cows screened, 49% (n = 130) were positive for bovine subclinical mastitis. One hundred thirty mastitic milk samples were subjected to bacterial culturing, and one hundred (76%) S. aureus isolates were identified based on phenotypic characters. Sixty-eight confirmed S. aureus isolates were obtained using PCR. The confirmed S. aureus isolates were tested for six virulence genes (tsst-1, hlb, eta, sea, clfA, and icaD) using PCR. Of the six virulence genes screened from all the isolates, only two (clfA and eta) were detected in the isolates. Out of 68 isolates, 25% and 22% were possessed the eta and clfA genes, respectively. Conclusion The presence of Staphylococcus aureus having virulence genes (eta and clfA) revealed that mastitis is a major concern nowadays affecting animal health, milk quality, and yield. Further genomic study of these isolates will provide broad new insights on virulence.

scholarly journals Study of Antibiotics Sensitivity Pattern And Molecular Characterization of Staphylococcus Aureus Isolated From Human And Animal Pyogenic Cases

2021 ◽  
Author(s):  
Jayshree Singh ◽  
Amit Kumar ◽  
Sharad K. Yadav ◽  
Ritika Yadav ◽  
Vinod K. Singh
Keyword(s):  
Staphylococcus Aureus ◽  
Molecular Characterization ◽  
Drug Sensitivity ◽  
Methicillin Resistance ◽  
Phenotypic Characterization ◽  
Resistance Pattern ◽  
Sensitivity Pattern ◽  
Drug Resistance Pattern ◽  
Animal Isolates

Abstract Staphylococcus aureus has been described as the most common cause of human and animal diseases and has emerged as superbug due to multidrug resistance. Considering these, a total of 175 samples were collected from pyogenic cases of humans (75) and animals (100), to establish the drug resistance pattern and also for molecular characterization of human and animal isolates. Thermonuclease (nuc) gene amplification was used to confirm all presumptive S. aureus isolates and then antibiotic sensitivity and slide coagulase tests were used for phenotypic characterization of isolates. Following that, all of the isolates were subjected to PCR amplification to detect the existence of the methicillin resistance (mecA) and coagulase (coa) genes. Lastly, typing was done by using the Randomly Amplified Polymorphic DNA-PCR. The overall prevalence of S. aureus in human and animal samples was found to be 39.4%. Drug sensitivity revealed the highest resistance against the β-lactam antibiotics such as ampicillin (94.8%) and penicillin (90.6%), followed by cephalosporin (cefixime-67.7%) and quinolone (ciprofloxacin-52.1%) group of drugs. The drug sensitivity was the highest against antibiotics like chloramphenicol (95%) followed by gentamicin (90%). Among the 69 S. aureus isolates, the overall presence of MRSA was 40.5% (27.5% and 50% in human and animal isolates respectively). Total 33 isolates exhibited coa genes amplification of more than one amplicons and variable in size of 250, 450, 800, and 1100 bp. The RAPD typing revealed amplification of 5 and 6 different band patterns in humans and animals, respectively, with two common patterns suggesting a common phylogenetic profile.

scholarly journals Within-Host Adaptation of Staphylococcus aureus in a Bovine Mastitis Infection Is Associated with Increased Cytotoxicity

2021 ◽  
Vol 22 (16) ◽  
pp. 8840
Author(s):  
Katharina Mayer ◽  
Martin Kucklick ◽  
Helene Marbach ◽  
Monika Ehling-Schulz ◽  
Susanne Engelmann ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Bovine Mastitis ◽  
Adaptive Strategy ◽  
Subclinical Mastitis ◽  
Growth Conditions ◽  
Typical Feature ◽  
Host Adaptation ◽  
Bovine Mammary Epithelial Cells ◽  
Strain Pair

Within-host adaptation is a typical feature of chronic, persistent Staphylococcus aureus infections. Research projects addressing adaptive changes due to bacterial in-host evolution increase our understanding of the pathogen’s strategies to survive and persist for a long time in various hosts such as human and bovine. In this study, we investigated the adaptive processes of S. aureus during chronic, persistent bovine mastitis using a previously isolated isogenic strain pair from a dairy cow with chronic, subclinical mastitis, in which the last variant (host-adapted, Sigma factor SigB-deficient) quickly replaced the initial, dominant variant. The strain pair was cultivated under specific in vitro infection-relevant growth-limiting conditions (iron-depleted RPMI under oxygen limitation). We used a combinatory approach of surfaceomics, molecular spectroscopic fingerprinting and in vitro phenotypic assays. Cellular cytotoxicity assays using red blood cells and bovine mammary epithelial cells (MAC-T) revealed changes towards a more cytotoxic phenotype in the host-adapted isolate with an increased alpha-hemolysin (α-toxin) secretion, suggesting an improved capacity to penetrate and disseminate the udder tissue. Our results foster the hypothesis that within-host evolved SigB-deficiency favours extracellular persistence in S. aureus infections. Here, we provide new insights into one possible adaptive strategy employed by S. aureus during chronic, bovine mastitis, and we emphasise the need to analyse genotype–phenotype associations under different infection-relevant growth conditions.

scholarly journals Risk factors associated with the occurrence of multiresistant Staphylococcus spp. isolated from bovine subclinical mastitis in northern Brazil

2022 ◽  
Vol 43 (2) ◽  
pp. 901-910
Author(s):  
Lilian Bernardina Ferreira ◽  
◽  
Larissa de Freitas Santiago Israel ◽  
Renata Fernandes Rabello ◽  
Guilherme Nunes de Souza ◽  
...  
Keyword(s):  
Risk Factors ◽  
Multidrug Resistance ◽  
Phenotypic Expression ◽  
Bovine Mastitis ◽  
Subclinical Mastitis ◽  
Isolation And Identification ◽  
Phenotypic Profile ◽  
Milk Samples ◽  
California Mastitis Test ◽  
Staphylococcus Spp

Staphylococcus bacteria are often associated with subclinical bovine mastitis. This study aimed to identify multiresistant Staphylococcus spp. associated with subclinical mastitis and the associated risk factors. Twenty-three dairy farms with a history of decrease in milk production, located in the lower Acre region, Brazil, were selected. An epidemiological questionnaire was provided in all farms. All animals were examined using the California Mastitis Test (CMT) and their milk samples were collected for bacterial culture. After isolation and identification, the disk diffusion antimicrobial susceptibility test was performed against nine classes of antimicrobials. Of the 339 cows examined using the CMT, 108 had mastitis. A total of 229 milk samples were collected from individual teats. MALDI-TOF MS found isolates belonging to eight species of Staphylococcus, in 101 of these samples. S. chromogenes (58.4%) demonstrated strongest resistance to the nine classes of antimicrobial active principles. Nineteen isolates with multidrug resistance phenotypic profile were identified. This phenotypic expression indicates wide circulation of resistant genes in this species. The presence of multidrug resistance in Staphylococcus spp. in this study was correlated with lack of water for cleaning the corral, which is a preventive factor, minimizing the transmission and persistence of pathogens in the farms.

Sign in / Sign up

Export Citation Format

Share Document