Enterotoxigenic methicillin resistant Staphylococcus aureus contamination in salted fish from Gwadar Balochistan

Staphylococcus aureus is an important foodborne pathogen associated to food intoxication and other multiple infections in human being. Its presence in salted food is a serious issue due to its salt tolerance potential. A study was conducted to analyze the presence of enterotoxins producing drug resistance S. aureus in salted sea fish from Gwadar. Freshly persevered samples (n=50) of salted fish were subjected to analyze the presence of S. aureus using 16S rRNA and Nuc genes primers. The isolates were then evaluated for drug resistance and enterotoxins producing potential using specific primers for MecA (methicillin resistance gene), (SEA) staphylococcal enterotoxin A and (SEB) staphylococcal enterotoxin B genes. Total 13/50 (26%) of the samples were found positive for the presence of S. aureus, preliminary confirmed with biochemical profiling and finally with the help of target genes presence. The isolates were found showing 100% resistant to methicillin, which were molecularly confirmed by the presence of MecA gene present in genome. The isolates 5/13 (38%) were positive for SEA and 3/13 (23%) for SEB genes, whereas 2/13 (15%) were confirmed having both SEA and SEB genes in its genome. It was also confirmed that all the isolates were capable to form biofilm over the glass surfaces. It was concluded that the study confirmed the presence of enterotoxigenic methicillin resistance Staphylococcus aurous (MRSA) in salted fish product, that poses gross food safety concern. Preventive and control measures are necessary to handle this serious food safety concern.

Classification of dairy properties according to the geometric mean of the somatic cell count and its antimicrobial resistance profile

Milk and its derivatives are highly consumed foods worldwide, with recognized nutritional importance. The search for the production of products with superior quality is constant. For the present work, 26 milk-producing properties were selected, with a total of 506 milk samples collected during the period from October 2019 to May 2020 being evaluated. The objective of this study was to evaluate the quality of milk produced in dairy properties in the region west Paraná, classified as good or bad based on the results of the Somatic Cell Count (SCC) and through sampling (n = 10) to evaluate the resistance profile of enterobacteria and Staphylococcus spp. isolated from milk samples, in addition to the presence of the mecA gene in strains of Staphylococcus spp. resistant to oxacillin. There were significant differences between the good and bad properties for the levels of lactose, SCC (cell/mL), and Standard Plate Count (SPC) (CFU/mL). The strains of Staphylococcus spp. showed differences in the percentage of resistance in relation to the good and bad properties for antibiotics: tetracycline, ciprofloxacin, oxacillin, amikacin, clindamycin, gentamycin, and erythromycin. The mecA gene was not detected in any of the coagulase-negative Staphylococcus isolates that showed resistance to oxacillin. For enterobacteria, the isolated species differed in relation to the classification of properties, with predominance for Escherichia coli (40%) for properties classified as bad and Hafnia alvei (40%) for those classified as good. The percentage of antibiotic resistance compared to enterobacteria isolates was higher in properties classified as good. Monitoring through microbial culture and antibiogram is extremely important, favoring the correct choice for the treatment of animals with a reduced selection of resistant strains.

mecA positive Staphylococcus spp. in bovine mastitis, milkers, milking environment, and the circulation of different MRSA clones at dairy cows farms in the Northeast region of Brazil

ABSTRACT: This study detected the presence and distribution of mecA in Staphylococcus spp. in the dairy production environment at farm level in Brazil. We analyzed 335 samples of mastitis cow milk, 15 samples of nostrils and hand swabs from milkers, 14 teat cup swabs, and 9 milking buckets swabs. Initially, the samples were subjected to microbiological analysis to detect Staphylococcus spp. and then S. aureus and mecA positive isolates were identified by PCR. All S. aureus isolates carrying the mecA genes were subjected to DNA macro-restriction analysis by Pulsed-Field Gel Electrophoresis (PFGE). The mecA gene was detected in 6/335 (1.78%) of mastitis cow milk, 5/15 (33.3%), and 5/15 (33.3%) of nostrils and hand swab, and 4/14 (28.5%) of the teat cup isolates. MRSA genotyping was performed by PFGE, a total of seven pulsotypes were grouped in two clusters. This study identified the occurrence and spread of MRSA at dairy environment of farms, and also the existence of distinct genetic profiles between isolates.

Isolation and antimicrobial resistance of coagulase-negative staphylococci recovered from healthy tortoises in Minas Gerais, Brazil

ABSTRACT: In the last few decades, there has been a global increase in the adoption of reptiles as companion animals, mainly turtles and tortoises. Considering the popularity of reptiles as pets in Brazil, and a notable lack of data about potentially pathogenic staphylococci in these animals, this study isolated and evaluate the antimicrobial susceptibility of staphylococcal species from healthy tortoises (Chelonoidis carbonaria) in Brazil. During a 12-month period (February 2019 to February 2020), cloacal swabs from 66 healthy tortoises were collected at the Wild Animals Screening Center in Belo Horizonte, Minas Gerais, Brazil. The swabs were plated onto mannitol salt agar for staphylococci isolation, and species identification was performed using MALDI-TOF MS. Antimicrobial susceptibility was investigated using the disk diffusion method, and the presence of the mecA gene was investigated by PCR to detect methicillin resistance. Of the tested animals, 72.7% were positive for staphylococcal isolation. All isolates were coagulase-negative staphylococci (CoNS), and Staphylococcus sciuri (81.3%), and S. xylosus (12.5%) were the most frequently isolated species. The majority of the isolates (56%) were resistant to at least one antimicrobial agent. A high frequency of resistance was observed for penicillin (35.5%) and tetracycline (29.1 %). All strains were susceptible to cefoxitin, chloramphenicol, ciprofloxacin, erythromycin, and gentamicin. All isolates were negative for the mecA gene. The present work suggests that healthy tortoises are mainly colonized by CoNS, especially S. sciuri. Half of the isolates were resistant to at least one antimicrobial, raising questions regarding the possible role of these animals as reservoirs of antimicrobial resistance genes.

Antimicrobial Resistance of Staphylococcus sp. Isolated from Cheeses

S. aureus and some species of coagulase-negative staphylococci, including S. chromogenes and S. simulans, commonly cause intramammary infections. However, little attention was paid to the antimicrobial resistance of these species with respect to their occurrence in dairy products, for example, popular sheep and goat cheeses made from unpasteurized milk. The aim of this study was to investigate such sheep and goat cheeses for the occurrence and antimicrobial resistance of the relevant staphylococci species. The staphylococcal isolates were identified by polymerase chain reaction (130 isolates) and matrix assisted laser desorption/ionization time-of-flight mass spectrometry. The most common species of S. aureus (56 isolates) were identified, as well as S. chromogenes (16 isolates) and S. simulans (10 isolates). Antimicrobial resistance to penicillin, oxacilin, ceftaroline, teicoplanin, gentamicin, erythromycin, tetracycline and ofloxacin was subsequently determined in these species using the agar dilution method. The highest resistance was confirmed in all species, especially to penicillin (91%) and erythromycin (67%). The highest sensitivity was confirmed to ofloxacin (83%). Due to the high incidence of penicillin and oxacilin-resistant staphylococci, the mecA gene was detected by polymerase chain reaction, which was confirmed only in S. aureus isolates (19%). Our study shows that the tested strains (77%) were resistant to more than one antibiotic at a time.

A comparative characterization of nasal and clinical isolates of Staphylococcus aureus from west of Iran

Background and Objectives: Recently, the rise of methicillin-resistant Staphylococcus aureus (MRSA) isolated from hos- pital healthcare workers (HCWs) and various infectious samples has become one of the main concerns in hospital settings. Therefore, epidemiological studies are necessary to monitor antibiotic resistance patterns in each region and to study the pathogenesis of this strain to control infections. Materials and Methods: In this cross-sectional study, a total of 100 S. aureus isolates, including 50 isolates obtained from the anterior nares of healthcare workers, as well as 50 other isolates cultured from the various clinical specimens from the referral hospitals in Khorramabad (West of Iran) were tested. All isolates were examined to determine antibiotic resistance pattern, and the presence of staphylococcal enterotoxin A (sea), staphylococcal enterotoxin B (seb) and mecA genes. Results: The mecA gene was found among 36% (18/50) of the clinical S. aureus isolates (CSIs) and 14% (7/50) of nasal S. aureus isolates (NSIs), with statistically significant difference (X2 = 6.53; p = 0.011). The difference between the frequency rate of sea gene among MRSA strains isolated from clinical specimens (46.6%, 7/15) was significant compared to strains isolated from nostrils (14.3%, 1/7) (X2 = 3.85; p = 0.049). Conclusion: The frequency of mecA, sea, and seb genes among the clinical samples was more than strains isolated from the nostrils of healthcare personnel.

ANTIBIOTIC RESISTANCE, MECA GENE DETECTION, AND BIOFILM FORMATION ABILITY AMONG COAGULASE-NEGATIVE STAPHYLOCOCCI IN CANCER PATIENTS

Objective: The objective of the study was to identify coagulase-negative staphylococci (CoNS) from various clinical samples and to determine the antibiotic resistance of the isolates by means of automation (VITEK-2), as well as to detect biofilm formation using Congo red agar method and to detect mecA gene by automated identification method (VITEK-2). Methods: All the clinical samples (blood, urine, sputum, BAL, throat swab, wound swab, aspirated fluid, pleural fluid, and pus) received in the microbiology laboratory were processed by aseptic techniques. Clinical samples were inoculated on appropriate media (blood agar, MacConkey agar, and chocolate agar [HIMEDIA]). After inoculation, the culture plates were incubated at 37°C aerobically for 18–24 h for growth. Positive cultures were picked up and further bacterial species identification was done using automated techniques (MALDI- TOF). Results: Among 28 isolates, the most recurrent strains of CoNS are Staphylococcus epidermidis, Staphylococcus hominis, Staphylococcus lugdunensis, and Staphylococcus haemolyticus. The assessment of antibacterial drugs sensitivity shows that all the isolates were more sensitive to daptomycin (S. epidermidis 100%, S. hominis 100%, S. lugdunensis 100%, and S. haemolyticus 42.85%) followed by linezolid (S. epidermidis [69.23%], S. hominis [100%], S. lugdunensis [100%], and S. haemolyticus [57.14%]) and vancomycin (S. epidermidis [100%], S. hominis [40%], S. lugdunensis [100%], S. haemolyticus [42.85%]). The analysis revealed the presence of the mecA gene (67.85%) and biofilm production (85.71%), respectively. Conclusion: Our data indicate that the hospital environment can be colonized by biofilm forming CoNS and transmission of these strains can cause an increased risk of serious nosocomial infections.

Prevalence and Patterns of Antimicrobial Resistance among Escherichia coli and Staphylococcus spp. in a Veterinary University Hospital

The occurrence of antimicrobial resistance in commensal strains of Escherichia coli and Staphylococcus spp. was investigated in 320 samples collected from patients and the environment of a veterinary university hospital—specifically, the consultation area (CA) and intensive care unit (ICU). E. coli was isolated in 70/160 samples (44%), while Staphylococcus spp. were isolated in 110/160 (69%) samples. The occurrence of multidrug-resistant (MDR) isolates from CA and ICU admission were similar for E. coli (1/12 (8%) versus 4/27 (15%), respectively) and Staphylococcus spp. (10/19 (53%) versus 26/50 (52%), respectively). MDR E. coli isolates increased significantly at hospital discharge (18/31; 58%; p = 0.008). Antimicrobial treatment administered during hospitalization was a risk factor for carriage of MDR E. coli (OR, 23.9; 95% CI: 1.18–484.19; p = 0.04) and MDR Staphylococcus spp. (OR, 19.5; 95% CI 1.30–292.76; p = 0.02), respectively. The odds ratio for MDR E. coli was 41.4 (95% CI 2.13–806.03; p = 0.01), if the administration of fluoroquinolones was evaluated. The mecA gene was detected in 19/24 (79%) coagulase-positive Staphylococcus spp. isolates resistant to oxacillin. High rates of MDR Staphylococcus spp. were reported. Hospitalization in the ICU and antimicrobial treatment were risk factors for colonization by MDR commensal bacteria.

Antibiotic resistance profiles of Staphylococcus spp. from white button mushrooms and handlers

The presence of Staphylococcus spp. has increasingly been reported in food products and poses a public health threat. The aim of this study was to determine the diversity of Staphylococcus spp. and the antibiotic resistance profiles of isolates obtained from freshly harvested and packed ready-to-eat mushrooms (n=432) and handlers’ hands (n=150). A total of 56 Staphylococcus isolates [46.4% (n=26) from hands and 53.6% (n=30) from mushrooms] were recovered belonging to 10 species. Staphylococcus succinus isolates (n=21) were the most prevalent, of which 52.4% came from mushrooms and 47.6% from hands. This was followed by S. equorum isolates [n=12; 91.7% (n=11) from mushrooms and 8.3% (n=1) from hands] and S. saprophyticus [n=9; 66.7% (n=6) from mushrooms and 33.3% (n=3) from hands]. Six isolates that were characterised as multidrug resistant were isolated from hands of handlers. Most (83.9%; n=47) of the 56 isolates were resistant to penicillin [53.2% (n=25) from mushrooms and 46.8% (n=22) from hands] and 14.3% (n=8) were resistant to cephalosporin classes [25% (n=2) from mushrooms and 75% (n=6) from hands], both of which are used to treat staphylococcal infections. Antibiotic resistance genes blaZ [25.0% (n=14) of all isolates of which 71.4% (n=10) were from hands and 28.57% (n=4) from mushrooms], tetL and tetK [both 1.8% (n=1) from hands], mecA [5.4% (n=3) from hands] and ermA [1.8% (n=1) from mushrooms] were detected from the 56 isolates. Only two (25.0%) of the eight methicillin-resistant staphylococci harboured the mecA gene, while only 11 (23%) of the 47 penicillin-resistant isolates harboured the blaZ gene [36.4% (n=4) from mushrooms and 63.6% (n=7) from hands]. Our results demonstrate that food handlers and harvested and packed ready-to-eat mushrooms could be a source of diverse Staphylococcus spp. that exhibit antimicrobial resistance. Clinically relevant S. aureus was only detected on one handler’s hand; however, the isolate was not multidrug resistant. The presence of diverse Staphylococcus spp. on mushrooms and the hands of handlers is a potential public health concern due to their potential to cause opportunistic infections.