scholarly journals Occurrence of Methicillin-Resistant Coagulase-Negative Staphylococci (MRCoNS) and Methicillin-Resistant Staphylococcus aureus (MRSA) from Pigs and Farm Environment in Northwestern Italy

Antibiotics ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 676
Author(s):  
Miryam Bonvegna ◽  
Elena Grego ◽  
Bruno Sona ◽  
Maria Cristina Stella ◽  
Patrizia Nebbia ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Human Health ◽  
Environmental Samples ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Bacterial Species ◽  
Meca Gene ◽  
Nucleotide Sequencing ◽  
Coagulase Negative Staphylococci ◽  
Methicillin Resistant ◽  
Nasal Swabs

Swine farming as a source of methicillin-resistant Staphylococcus aureus (MRSA) has been well documented. Methicillin-resistant coagulase-negative staphylococci (MRCoNS) have been less studied, but their importance as pathogens is increasing. MRCoNS are indeed considered relevant nosocomial pathogens; identifying putative sources of MRCoNS is thus gaining importance to prevent human health hazards. In the present study, we investigated MRSA and MRCoNS in animals and environment in five pigsties in a high farm-density area of northwestern Italy. Farms were three intensive, one intensive with antibiotic-free finishing, and one organic. We tested nasal swabs from 195 animals and 26 environmental samples from three production phases: post-weaning, finishing and female breeders. Phenotypic tests, including MALDI-TOF MS, were used for the identification of Staphylococcus species; PCR and nucleotide sequencing confirmed resistance and bacterial species. MRCoNS were recovered in 64.5% of nasal swabs, in all farms and animal categories, while MRSA was detected only in one post-weaning sample in one farm. The lowest prevalence of MRCoNS was detected in pigs from the organic farm and in the finishing of the antibiotic-free farm. MRCoNS were mainly Staphylococcus sciuri, but we also recovered S. pasteuri, S. haemolyticus, S. cohnii, S. equorum and S. xylosus. Fifteen environmental samples were positive for MRCoNS, which were mainly S. sciuri; no MRSA was found in the farms’ environment. The analyses of the mecA gene and the PBP2-a protein highlighted the same mecA fragment in strains of S. aureus, S. sciuri and S. haemolyticus. Our results show the emergence of MRCoNS carrying the mecA gene in swine farms. Moreover, they suggest that this gene might be horizontally transferred from MRCoNS to bacterial species more relevant for human health, such as S. aureus.

Prevalence of Methicillin-Resistant Staphylococcus aureus in a Fresh Meat Pork Production Chain

2011 ◽  
Vol 74 (1) ◽  
pp. 126-129 ◽  
Author(s):  
BIRGIT BENEKE ◽  
SYLVIA KLEES ◽  
BIRGIT STÜHRENBERG ◽  
ALEXANDRA FETSCH ◽  
BRITTA KRAUSHAAR ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Environmental Samples ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Processing Unit ◽  
Production Chain ◽  
Methicillin Resistant ◽  
Pork Production ◽  
Nasal Swabs ◽  
Meat Samples ◽  
Spa Types

The objective of this study was to investigate the prevalence of methicillin-resistant Staphylococcus aureus on different stages of a fresh pork production chain to reveal potential carryover from live animals to meat. Samples were collected at different stages of the production process in a large German abattoir with an integrated processing unit for fresh pork. Samples included nasal swabs from pigs at stunning, environmental samples from the slaughter line, surface samples from carcasses, environmental and meat samples from the processing unit, and samples from final products. Samples were analyzed with an established two-step selective enrichment method, and isolates were characterized with respect to their S. aureus protein A gene (spa) and staphylococcal cassette chromosome mec (SCCmec; which harbors the mecA gene) types. Contamination rate was highest (64.7%) in nasal swabs and lower (6.0%) on carcasses, meat at processing (4.2%), and final products (2.8%). Environmental samples were positive along the slaughter line (12%) but not in the processing unit. spa types t011 and t034 and SCCmec type V predominated the isolates. Heterogeneity of spa types was highest in nasal swabs. Results show that methicillin-resistant S. aureus can be identified at all stages of the production chain. Further studies are needed to identify potential control points to reduce the carryover from farm animals to the final products.

Longitudinal Study of Clostridium difficile and Methicillin-Resistant Staphylococcus aureus Associated with Pigs from Weaning through to the End of Processing

2013 ◽  
Vol 76 (4) ◽  
pp. 624-630 ◽  
Author(s):  
PHILIP HAWKEN ◽  
J. SCOTT WEESE ◽  
ROBERT FRIENDSHIP ◽  
KEITH WARRINER
Keyword(s):  
Staphylococcus Aureus ◽  
Clostridium Difficile ◽  
Environmental Samples ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Cross Contamination ◽  
Methicillin Resistant ◽  
Nasal Swabs ◽  
Processing Facility ◽  
Ribotype 078 ◽  
Spa Type

There has been a recent increase in community-associated infections linked to methicillin-resistant Staphylococcus aureus (MRSA) and Clostridium difficile. It is established that both pathogens can be recovered from retail pork, although it is unclear to what degree contamination is acquired at the farm in comparison to that acquired during processing. To address this gap, the following study reports on the carriage of MRSA and C. difficile on pigs from birth through to the end of processing. C. difficile was isolated from 28 (93%) of 30 pigs at 1 day of age, but prevalence declined sharply to 1 of 26 by market age (188 days). MRSA prevalence peaked at 74 days of age, with 19 (68%) of 28 pigs testing positive, but declined to 3 of 26 at 150 days of age, with no pig being detected as positive at market age. At the processing facility, C. difficile was isolated from the holding area, with a single carcass testing positive for the pathogen at preevisceration. MRSA was primarily isolated from nasal swabs with 8 (31%) carcasses testing positive at postbleed, which increased to 14 (54%) positive at postscald tanks. Only one carcass (sampled at postbleed) tested positive for MRSA, with no recovery of the pathogen from environmental samples taken. C. difficile ribotype 078 predominated in the longitudinal portion of the study, accounting for all of the 68 isolates recovered from pigs. Only three C. difficile isolates, which were identified as ribotype 078, were recovered at the slaughterhouse. MRSA spa type 539 (t034) predominated in pigs on the farm and samples taken at the slaughterhouse, accounting for 80% of all isolates recovered. The study demonstrated that both C. difficile and MRSA acquired on the farm can be transferred through to processing, although no evidence for significant cross-contamination between carcasses or the slaughterhouse environment was evident.

scholarly journals Molecular detection and typing of methicillin-resistant Staphylococcus aureus and methicillin-resistant coagulase-negative staphylococci isolated from cattle, animal handlers, and their environment from Karnataka, Southern Province of India

2019 ◽  
Vol 12 (11) ◽  
pp. 1760-1768 ◽  
Author(s):  
Nimita Venugopal ◽  
Susweta Mitra ◽  
Rituparna Tewari ◽  
Feroze Ganaie ◽  
Rajeswari Shome ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Methicillin Resistance ◽  
Animal Health ◽  
Sccmec Type ◽  
Meca Gene ◽  
Molecular Characteristics ◽  
Rrna Gene ◽  
Coagulase Negative Staphylococci ◽  
Methicillin Resistant

Background and Aim: Methicillin-resistant staphylococci are among the emerging pathogens which have become a threat to both human and animal health. The present investigation intended to examine the occurrence and the molecular characteristics of methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-resistant coagulase-negative staphylococci (MRCoNS) recovered from cattle, its handlers, and their environment. Materials and Methods: A total of 666 specimens were subjected to culture method and genus-specific polymerase chain reaction (PCR) for the identification of Staphylococcus. Methicillin resistance was substantiated by PCR identification of mecA and mecC resistance determinants. Species-specific identification of mecA positive isolates was conducted by multiplex PCR. The unidentified species were deciphered by 16S rRNA gene sequencing approach. The mecA positive isolates were further characterized by staphylococcal cassette chromosome mec (SCCmec) typing and multilocus sequence typing (MLST). Results: Duplex PCR identified 728 Staphylococcus isolates, of which 66 (9%) were positive for mecA gene. MRSA constituted 24% of the total mecA positive isolates. Among MRCoNS, Staphylococcus epidermidis (42%), and Staphylococcus haemolyticus (11%) were the most common species identified. Overall, 47% of the mecA positive isolates belonged to SCCmec type V. MLST analysis showed eight different sequence types (STs) among MRSA isolates of which five were novel STs. Among methicillin-resistant S. epidermidis, 19 different STs were found, of which nine novel STs were detected. Conclusion: The increase in the prevalence of mecA positive staphylococci, especially MRCoNS in cattle is a great concern in view of their transmission potential. Hence, continuous monitoring and molecular characterization of methicillin-resistant staphylococci should be elucidated in human and animal sectors so as to prevent the spread of these resistant pathogens.

DETECTION OF METHICILLIN-RESISTANT STAPHYLOCOCCUS AUREUS (MRSA) FROM ANIMAL AND HUMAN ORIGIN IN BANGLADESH BY POLYMERASE CHAIN REACTION

2013 ◽  
Vol 9 (2) ◽  
pp. 161-166 ◽  
Author(s):  
MM Alam ◽  
MS Uddin ◽  
N Kobayashi ◽  
MU Ahmed
Keyword(s):  
Staphylococcus Aureus ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Bacterial Species ◽  
Meca Gene ◽  
Penicillin Binding Protein ◽  
Chain Reaction ◽  
Veterinary Clinic ◽  
Methicillin Resistant ◽  
Polymerase Chain ◽  
Animal Isolates

Methicillin-resistant Staphylococcus aureus (MRSA) is defined by the presence of the mecA gene, which is considered to have been transferred horizontally from unknown bacterial species to S. aureus. The mecA gene which encodes an additional ?-lactam-resistant penicillin-binding protein (PBP), termed PBP-2a (PBP-2’) with reduced binding affinity for ?-lactam compounds. We investigated distribution of the mecA gene in a total of 94 clinical strains of S. aureus isolated from both man and animal admitted in Bangladeshi medical hospital as well as Veterinary clinic. The mecA gene was detected by PCR in 25% of human clinical isolates of S. aureus, whereas not a single mecA gene was detected in animal isolates of S. aureus.DOI: http://dx.doi.org/10.3329/bjvm.v9i2.13472

scholarly journals Occurrence and characteristics of methicillin resistant Staphylococcus aureus and methicillin resistant coagulase-negative staphylococci in raw milk manufacturing

2012 ◽  
Vol 29 (Special Issue) ◽  
pp. S11-S16 ◽  
Author(s):  
M. Vyletělová ◽  
H. Vlková ◽  
I. Manga
Keyword(s):  
Staphylococcus Aureus ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Methicillin Resistance ◽  
Meca Gene ◽  
Diffusion Method ◽  
Biochemical Tests ◽  
Coagulase Negative Staphylococci ◽  
Disk Diffusion Method ◽  
Methicillin Resistant ◽  
Staphylococcal Cassette Chromosome

For monitoring the occurrence of MRSA (methicillin resistant Staphylococcus aureus) and MR-CNS (methicillin resistant coagulase-negative staphylococci), cow’s, goat’s, and sheep’s milks (bulk milks and individual samples) were investigated. Human nasal and throat swabs of the farm staff and nasal swabs of animals were also investigated as well. In total 1729 samples were examined and 634 strains were isolated by means of the cultivation method and used in this study. Generic identification of the staphylococci isolates was done performed by biochemical tests and all S. aureus and CNS isolates were checked by the PCR method for the presence of mecA gene which is responsible for methicillin resistance. The presence of the staphylococcal cassette chromosome mec (SCCmec), Panton-Valentine leukocidin (pvl) and genes encoding toxic shock syndrome toxin (tst) was detected in all strains confirmed as MRSA. The species were also examined for antimicrobial susceptibility by using disk diffusion method with antibiotic disks. S. aureus was the most frequently identified species from the samples tested (n = 557; 32.2%), followed by S. haemolyticus (n = 32; 1.9%), S. chromogenes (n = 24; 1.4%), S. epidermidis (n = 20; 1.2%), and S. caprae (n = 1; 0.16%). Among the resistant staphylococci (n = 49), S. aureus (n = 25; 51%) was found the most frequently, followed by S. epidermidis (n = 17; 34.7%), S. chromogenes (n = 6; 12.2%), and S. haemolyticus (n = 1; 2%). The resistant Staphyloccocus sp. occurred mainly in cow’s milk (MRSA, S. epidermidis, S. chromogenes, S. haemolyticus) and in animal’s swabs (S. epidermidis). One MRSA was also found in goat’s milk and one was isolated from human swab. No resistant strains were found in sheep’s milk. The negative results of the analysed genes presence (pvl, tst) were identical with all MRSA tested. The staphylococcal cassette chromosome mec (SCCmec) was classified as type IV or V.

scholarly journals Mutation-Based Antibiotic Resistance Mechanism in Methicillin-Resistant Staphylococcus aureus Clinical Isolates

2021 ◽  
Vol 14 (5) ◽  
pp. 420
Author(s):  
Tanveer Ali ◽  
Abdul Basit ◽  
Asad Mustafa Karim ◽  
Jung-Hun Lee ◽  
Jeong-Ho Jeon ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Antibiotic Resistance ◽  
Active Site ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Meca Gene ◽  
Resistance Mechanism ◽  
Ligand Docking ◽  
Clinical Samples ◽  
Allosteric Site ◽  
Methicillin Resistant

β-Lactam antibiotics target penicillin-binding proteins and inhibit the synthesis of peptidoglycan, a crucial step in cell wall biosynthesis. Staphylococcus aureus acquires resistance against β-lactam antibiotics by producing a penicillin-binding protein 2a (PBP2a), encoded by the mecA gene. PBP2a participates in peptidoglycan biosynthesis and exhibits a poor affinity towards β-lactam antibiotics. The current study was performed to determine the diversity and the role of missense mutations of PBP2a in the antibiotic resistance mechanism. The methicillin-resistant Staphylococcus aureus (MRSA) isolates from clinical samples were identified using phenotypic and genotypic techniques. The highest frequency (60%, 18 out of 30) of MRSA was observed in wound specimens. Sequence variation analysis of the mecA gene showed four amino acid substitutions (i.e., E239K, E239R, G246E, and E447K). The E239R mutation was found to be novel. The protein-ligand docking results showed that the E239R mutation in the allosteric site of PBP2a induces conformational changes in the active site and, thus, hinders its interaction with cefoxitin. Therefore, the present report indicates that mutation in the allosteric site of PBP2a provides a more closed active site conformation than wide-type PBP2a and then causes the high-level resistance to cefoxitin.

scholarly journals Isolation of Biofilm Producing Methicillin-Resistant Staphylococcus aureus from Hospitalized Orthopaedic Patients in Kano State, Nigeria

2021 ◽  
Vol 28 (1) ◽  
pp. 66-74
Author(s):  
D.A. Oche ◽  
U. Abdulrahim ◽  
A.S. Oheagbulem ◽  
B.O. Olayinka
Keyword(s):  
Staphylococcus Aureus ◽  
Biofilm Formation ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Methicillin Resistance ◽  
Meca Gene ◽  
Community Settings ◽  
Potential Threat ◽  
Orthopaedic Patients ◽  
Nasal Swabs ◽  
Public Health Challenge

Biofilm formation and resistance to methicillin are among the factors that makes Staphylococcus aureus a very important human pathogen in both health-care and community settings. This study investigated methicillin-resistance among biofilm-producing S. aureus isolated from 49 orthopaedic in-patients within a 3 months period. Wound swabs, nasal swabs, bed swabs and urine samples were collected from each patient. The samples were cultured and screened for presence of S. aureus while the micro-titre plate method was used to detect biofilm producing isolates. PCR technique was finally used to detect the presence of mecA gene in methicilin resistant S. aureus (MRSA) isolates. Findings reveal 14.8% of bacterial isolates were Staphylococcus aureus of which 96.4% were biofilm-producers. However, strong biofilm producers constitute 11.1%. The mecA gene was detected in 15.8% of the MRSA isolates. Therefore, MRSA among biofilm-producing S. aureus is a potential threat primarily to the community of National Orthopaedic Hospital Dala and a major public health challenge. Keywords: Biofilm, Methicillin-resistance Staphylococcus aureus (MRSA), mecA gene, Orthopaedic patient

scholarly journals Detection of mecA gene by latex agglutination and its molecular analysis by PCR in methicillin resistant staphylococcus aureus.

2020 ◽  
Vol 27 (07) ◽  
pp. 1363-1370
Author(s):  
Aneela Khawaja ◽  
Iffat Javed ◽  
Sohaila Mushtaq ◽  
Saeed Anwar ◽  
Faiqa Arshad ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Methicillin Resistance ◽  
Meca Gene ◽  
Latex Agglutination ◽  
Medical Institute ◽  
Cross Sectional ◽  
Methicillin Resistant ◽  
Agglutination Method ◽  
Resistant Strains

Antimicrobial resistance (AMR) is a devastating question that is threatening the health globally. The extensive and indiscriminative use of antibiotics has evolved a notorious resistance in Staphylococcus aureus.  This resistance developed through possession of mecA gene, which codes for modified penicillin binding protein (PBP2a) and the emergent strain being labeled “methicillin resistant Staphylococcus aureus”. Conventional phenotypic techniques for detection of MRSA rely on standardization of cultural characteristics. The drawbacks of diagnostic error to report MRSA include: poor prognosis, expensive treatment, dissemination of multi-drug resistant strains and even treatment failure. Latex agglutination method can be adopted as a more accurate and quick strategy for rapid detection of methicillin resistance. Objectives: To compare detection of mecA gene in methicillin resistant isolates of Staphylococcus aureus by latex agglutination and PCR; by assessing the sensitivity and specificity of both methods. Study Design: Descriptive Cross-Sectional study. Setting: Pathology Department, Post Graduate Medical Institute, Lahore. Period: From January 2015 to December 2015; according to standard operating procedures at Microbiology laboratory. Material & Methods: A total 713 consecutive, non-duplicate isolates of Staphylococcus aureus were processed. Methicillin resistance was determined using cefoxitin (30mg) by Kirby-Bauer method using CLSI guideline (2016), latex agglutination method; and PCR for mecA gene. Results: The results showed that out of 713 Staphylococcus aureus isolates, 92 (12.90%) isolates were resistant to cefoxitin and were labelled as MRSA. majority MRSA isolates recovered from pus (44.57%) and wound swab (20.65%), followed by blood (13.04%), fluid (8.70%), CSF (4.35%), CVP (3.26%), HVS (3.26%) and tracheal secretion (2.17%). By latex agglutination method, 87 (94.50%) were positive for PBP2a; while on PCR mecA gene was detected only in 82 (89.10%) MRSA isolates. When assessed with PCR (gold standard) the sensitivity and diagnostic accuracy of latex agglutination was 100% and 94.57%, respectively. Conclusion: Latex agglutination test can be employed as rapid and reliable diagnostic technique in MRSA isolates for mecA gene detection, where resources for molecular methods are inadequate. This can effectually lessen the misdiagnosis of resistant strains, and over/ ill-use of antibiotics.

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