Occurrence, antimicrobial susceptibility, and resistance genes of Staphylococcus aureus in milk and milk products in the Arsi highlands of Ethiopia.

BACKGROUND: In Ethiopia, milk production and handling practices often lack proper hygiene measures, leading to the potential contamination of milk and milk products with Staphylococcus aureus (S. aureus), including methicillin-resistant strains, posing significant public health concerns. This study aimed to investigate the occurrence, antimicrobial susceptibility profiles and presence of resistance genes in S. aureus strains isolated from milk and milk products. METHODS: A cross-sectional study was conducted in the Arsi highlands, Oromia, Ethiopia from March 2022 to February 2023. A total of 503 milk and milk product samples were collected, comprising 259 raw milk, 219 cottage cheese, and 25 traditional yogurt samples. S. aureus isolation and coagulase-positive staphylococci enumeration were performed using Baird-Parker agar supplemented with tellurite and egg yolk. S. aureus was further characterized based on colony morphology, Gram stain, mannitol fermentation, catalase test, and coagulase test. Phenotypic antimicrobial resistance was assessed using the Kirby-Bauer disc diffusion method, while the polymerase chain reaction (PCR) was employed for confirming the presence of S. aureus and detecting antimicrobial resistance genes. RESULTS: S. aureus was detected in 24.9% of the milk and milk products, with the highest occurrence in raw milk (40.9%), followed by yogurt (20%), and cottage cheese (6.4%). The geometric mean for coagulase-positive staphylococci counts in raw milk, yogurt, and cottage cheese was 4.6, 3.8, and 3.2 log10 CFU/mL, respectively. Antimicrobial resistance analysis revealed high levels of resistance to ampicillin (89.7%) and penicillin G (87.2%), with 71.8% of the isolates demonstrating multidrug resistance. Of the 16 S. aureus isolates analyzed using PCR, all were found to carry the nuc gene, with the mecA and blaZ genes detected in 50% of these isolates each. CONCLUSION: This study revealed the widespread distribution of S. aureus in milk and milk products in the Arsi highlands of Ethiopia. The isolates displayed high resistance to ampicillin and penicillin, with a concerning level of multidrug resistance. The detection of the mecA and blaZ genes in selected isolates is of particular concern, highlighting a potential public health hazard and posing a challenge to effective antimicrobial treatment. These findings highlight the urgent need to enhance hygiene standards in milk and milk product handling and promote the rational use of antimicrobial drugs. Provision of adequate training for all individuals involved in the dairy sector can help minimize contamination. These measures are crucial in addressing the threats posed by S. aureus, including methicillin-resistant strains, and ensuring the safety of milk and its products for consumers.

The polymicrobial nature of the oral cavity and claws of cats diagnosed by mass spectrometry and next-generation sequencing.

Close contact between cats and humans increases the risk of transmission of zoonotic pathogens, through bites and scratches due to the complexity of microorganisms in the oral and nail microbiotas of felines. This study investigated the presence of bacteria and fungi in the oral cavity and claws of 100 apparently healthy cats using conventional and selective microbiological culture media, and next-generation sequencing (NGS) and mass spectrometry (MALDI-TOF MS). Furthermore, antimicrobial susceptibility testing of bacteria isolates was performed by disc diffusion method. In total, 671 bacteria and 33 yeasts were identified by MALDI-TOF MS. Neisseria animaloris (10.8 %), Staphylococcus felis (8.5 %), and Pasteurella multocida (7 %) were the most prevalent bacteria in oral cavity samples (n = 343), while the most common yeast (n = 19) was Candida albicans (68.4 %). Staphylococcus pettenkoferi (13.4 %), Staphylococcus felis (6.4 %), and Staphylococcus simulans (5.8 %) were the prevalent bacteria identified in the claw samples (n = 328), while Rhodotorula mucilaginosa (57.2 %) was the most common yeast (n = 14). NGS predominantly identified the genera Moraxella, Neisseria, Pasteurella, and Fusobacterium in oral cavity samples, whereas enterobacteria and staphylococci were prevalent in nail bed samples. In addition, the genera Capnocytophaga and Bartonella were identified, which have been described in serious human infections secondary to feline aggressions. Levofloxacin, marbofloxacin, and amoxicillin/clavulanic acid were the most effective drugs against the main groups of bacteria identified. Multidrug resistance was observed in 17 % of the bacterial isolates. Furthermore, three staphylococci harboring the methicillin resistance gene mecA were identified. We highlight the complexity of microorganisms inhabiting the oral/claw microbiotas of cats, the high resistance rate of the isolates to conventional antimicrobial agents, and the zoonotic risk of aggressions caused by bites and scratches from domestic cats.

A simple and sensitive colorimetric approach for mecA gene analysis via exonuclease-III catalyzed signal cascade.

Analysis of mecA gene in Staphylococcus aureus (S. aureus) is essential for controlling infections in intensive care units (ICU) and preventing the use of ineffectual empirical treatments. However, quantitative determination of the mecA gene remains difficult. Herein, we propose a simple and sensitive colorimetric approach by integrating exonuclease-III (Exo-III) assisted signal cascade and G-quadruplex/hemin DNAzymes (G4 DNAzymes) catalyzed 2,2'-azino-bis (3-ethylben-zothiazoline-6-sulfonic acid) (ABTS) based color reaction. In this method, signal amplification does not necessitate the use of complex experimental components, such as multiple enzymes and primer design, while still maintaining a high signal amplifying efficiency. Therefore, the method has a broad mecA gene detection range from 10 fM to 1 nM and a low limit of detection down to 3.4 fM level. Taking the merit of simplicity and high sensitivity, the approach is promising in analyzing mecA gene in S. aureus and diagnosing infections.

Berberine hydrochloride reduces staphyloxanthin synthesis by inhibiting fni genes in methicillin-resistant Staphylococcus aureus.

BACKGROUND: Methicillin-resistant Staphylococcus aureus (MRSA) poses a great health threat to humans. Looking for compounds that could reduce the resistance of S. aureus towards methicillin is an effective way to alleviate the antimicrobial resistance crisis. METHODS AND RESULTS: Minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), Time-killing growth curve, staphyloxanthin and penicillin-binding protein 2a (PBP2a) were detected. A quantitative polymerase chain reaction was used to measure the effect of BBH on the gene transcription profiles of MRSA. The MIC of MRSA-ST59-t437 towards oxacillin was 8 µg/ml, and MBC was 128 µg/ml. After adding a sub-inhibitory concentration of BBH, the MIC and MBC of MRSA-ST59-t478 towards oxacillin went down to 0.125 and 32 µg/ml respectively. The amount of PBP2a and staphyloxanthin were reduced after treatment with BBH. Moreover, the transcription levels of sarA, mecA and fni genes were downregulated. CONCLUSIONS: It is for the first time reported that BBH could inhibit staphyloxanthin synthesis by inhibiting fni gene. Moreover, fni might be the target gene of sarA, and there might be another regulatory pathway to inhibit staphyloxanthin biosynthesis. BBH could effectively reduce the methicillin resistance of MRSA-ST59-t437 by downregulating fni, sarA and mecA genes.

Failure of mecA/mecC PCR Testing to Accurately Predict Oxacillin Resistance in a Patient with Staphylococcus aureus Infective Endocarditis.

Genotypic testing for mecA/mecC is heavily relied upon for rapid optimization of antimicrobial therapy in infections due to Staphylococcus aureus. Little is known regarding optimal reporting and/or therapy for patients demonstrating lack of genotypic evidence of mecA or mecC but phenotypic oxacillin resistance. We report a case of a 77-year-old patient with S. aureus bloodstream infection and infective endocarditis with discordance between mecA/mecC genotypic results and phenotypic susceptibility testing.

Panton-Valentine Leukocidin (PVL) genes may not be a reliable marker for community-acquired MRSA in the Dakahlia Governorate, Egypt.

BACKGROUND: Methicillin-resistant Staphylococcus aureus is linked to both nosocomial and community infections. One of the key virulence factors of S. aureus is Panton-Valentine leukocidin (PVL). The PVL genes are mostly associated with community-acquired MRSA (CA-MRSA). This study evaluates the prevalence of PVL genes as a marker for CA-MRSA at tertiary hospitals in Mansoura, Dakahlia, Egypt. S. aureus was isolated from clinical specimens obtained from different departments of tertiary hospitals, outpatient clinics, and hospital healthcare workers (HCWs). PCR was used to detect the mecA, PVL, and SCCmec genes among the recovered isolates. Standard broth microdilution method was used to determine the minimum inhibitory concentrations (MIC) of nine antibiotics against S. aureus. RESULTS: Two hundred S. aureus isolates were recovered and identified out of the total isolates (n = 320). The mecA gene was detected in 103 S. aureus isolates (51.5%). Among the MRSA isolates, 46.60% were PVL-positive. The incidence of the PVL genes of MRSA in nosocomial (HA), outpatient clinics (CA), and HCWs was 46.66%, 56.52%, and 42%, respectively. All MRSA isolates showed resistance to cefoxitin. The percentage of resistance to most tested antibiotics was high, except for ciprofloxacin (6.85%). Both antibiotic resistance and multidrug resistance among MRSA isolates were generally higher in PVL-positive isolates than in PVL-negative isolates in HA- and CA-MRSA isolates. While SCCmec type V was the most prevalent in PVL-positive MRSA stains, type I was the most prevalent in PVL-negative isolates. CONCLUSION: This study revealed that PVL genes are generally highly prevalent among mecA-positive MRSA isolates, whether they are CA-MRSA, HA-MRSA, or HCW isolates. Therefore, PVL is not a valid marker for CA-MRSA in Mansoura, Dakahlia Governorate, Egypt, as has been reported in other countries. Further epidemiologic studies are required to track the incidence of PVL in HA-MRSA, CA-MRSA, and HCW isolates in other Egyptian governorates.

Detection of virulence genes of Staphylococcus aureus isolated from raw beef for retail sale in the markets of Ulaanbaatar city, Mongolia.

BACKGROUND: Staphylococcus aureus (S. aureus) is a highly virulent pathogen that causes food-borne illness, food poisoning, skin and soft tissue infections, abscesses, mastitis, and bacteremia. It is common for meat and meat products to become contaminated with S. aureus due to dirty hands, food storage conditions, food production processes, and unhygienic conditions, causing food poisoning. Therefore, we aimed to isolate S. aureus strain from the raw beef and reveal virulence genes and antibiotic resistance profile from isolated S. aureus strains. METHODS: In this study, 100 samples of raw beef were collected from 4 major market stalls in Ulaanbaatar city, Mongolia. S. aureus was detected according to the ISO 6888-1:2021 standard, and the nucA gene encoding the species-specific thermonuclease was amplified and confirmed by polymerase chain reaction (PCR). In the strains of S. aureus isolated from the samples, the genes encoding the virulence factors including sea, sed, tsst, eta, etb, and mecA were amplified by multiplex PCR. These genes are encoded staphylococcal enterotoxin A, enterotoxin D, toxic shock syndrome toxin, exotoxin A, exotoxin B and penicillin-binding protein PBP 2A, respectively. Antibiotic sensitivity test was performed by the Kirby-Bauer disc diffusion method. The Clinical and Laboratory Standard Institute guidelines as CLSI M100-S27 was used for analysis of the data. RESULTS: Thirty-five percent of our samples were detected contaminated with of the S. aureus strains. Subsequently, antibiotic resistance was observed in the S. aureus contaminated samples. Among our samples, the highest rates of resistance were determined against ampicillin (97.1%), oxacillin (88.6%), and penicillin (88.6%), respectively. Three genes including mecA, sea, and tsst from six virulence genes were detected in 17% of S. aureus strain-contaminated samples by multiplex PCR. The sed, etb and eta genes were detected in the 2.9%, 11.4% and 5.7% of our samples, respectively. CONCLUSION: The results show that S. aureus related contamination is high in the raw beef for retail sale and prevalent S. aureus strains are resistant to all antibiotics used. Also, our results have demonstrated that there is a high risk for food poisoning caused by antibiotic resistant S. aureus in the raw beef and it may establish public health issues. Genes encoding for both heat-resistant and nonresistant toxicity factors were detected in the antibiotic resistant S. aureus strains and shown the highly pathogenic. Finally, our study is ensuring to need proper hygienic conditions during beef's preparation and sale.

Characterization and pathogenicity of multidrug-resistant coagulase-negative Staphylococci isolates in chickens.

The pathogenic potential of vancomycin and methicillin-resistant coagulase-negative Staphylococci (VMRCoNS) on Egyptian poultry farms has received little attention. Therefore, this study aims to study the prevalence of CoNS in imported poultry flocks and commercial poultry farms, evaluate the presence of virulence and antibiotic resistance genes (sea, seb, sec, sed, see, and mecA), and assess their pathogenicity in broiler chicks. Seven species were identified among 25 isolates, such as 8 S. gallinarum, 5 S. saprophyticus, 5 S. chromogens, 3 S. warneri, 2 S. hominis, 1 S. caprae, and 1 S. epidermidis. All isolates were resistant to clindamycin, doxycycline, vancomycin, methicillin, rifampicin, and penicillin. The mecA gene was confirmed in 14 isolates, while the sed gene was revealed in seven isolates. Commercial 1-day-old Ross broiler chicks were divided into eight groups of three replicates (10 birds/group): group Ӏ was negative control; groups (П, Ш, IV, V, VI, VII, and VIII) were subcutaneously inoculated with 108 CFUml-1 of S. hominis, S. caprae, S. epidermidis, S. gallinarum, S. chromogens, S. warneri, and S. saprophyticus, respectively. Groups VIII and V had mortality rates of 100% and 20%, respectively, with no evidence of mortalities in the other groups. The highest re-isolation of CoNS species was recorded in groups VII, VIII, and V. Postmortem and histopathological examination revealed the common presence of polyserositis in the internal organs, and hepatic and myocardial necrosis in groups IV, V, and VI. These findings revealed the pathogenic potential of CoNS, so special attention must be directed toward their public health impact.

Occurrence of virulence genes and methicillin-resistance in Staphylococcus aureus isolates causing subclinical bovine mastitis in Tiaret area, Algeria.

Mastitis remains the most frequent and the most expensive disease of dairy breeding. The objective of the study was to study S. aureus isolates collected from subclinical bovine mastitis in the Tiaret region, Algeria, by determining their antimicrobial susceptibility and their virulence traits. Sixty-two S. aureus isolates collected from subclinical bovine mastitis were studied by determining their antimicrobial susceptibility according to CLSI guidelines, and nine genes encoding virulence factors and resistance to methicillin and penicillin were determined by PCR. Multi-drug resistance was observed in 19 (30.64%) isolates and five (8%) were methicillin-resistant S. aureus (MRSA), four of them harbored the mecA gene; however, the mecC gene was not detected. Out of 59 penicillin-resistant isolates, 14 harbored the blaZ gene; one of them co-harbored the mecA gene. The following virulence genes were detected: eta (n = 23; 37%), icaA (20; 32.2%), icaD (18; 29%), etb (16; 25.8%), luk E-D (14; 22.5%), and sea (6; 9.6%). The tsst-1, lukF/lukS, and luk-M genes were not detected. The occurrence of MRSA and multi-drug resistant (MDR) S. aureus isolates as well as genes encoding virulence factors playing an important role in the pathogenesis of subclinical bovine mastitis and of harmful potential to human is a cause for concern.

Staphylococcal biofilm on wedding rings worn by laboratory workers.

Hands of healthcare workers play essential role in the spreading of antimicrobial-resistant microorganisms in and out of the healthcare settings. Less is known about the role of laboratory workers (LWs). The aim of our study was to evaluate the presence of biofilm-forming staphylococci on the surface of jewelry rings of LWs and their antimicrobial susceptibility pattern.A total of 79 LWs from eight different microbiology laboratories that process and analyze specimens from the tertiary care hospitals in Belgrade, Serbia participated in the study. The study was reviewed and approved by the institutional review boards at hospitals. Samples were taken after hand washing. Bacteria on LWs wedding rings were detected with the rolling method, and further analyzed in order to determine the number of colony forming unit (CFU) per ring, species of bacteria and their antimicrobial susceptibility pattern, methicillin resistance and biofilm-producing capacity in vitro.Staphylococci were recovered from 60.8% of wedding rings. All strains produced biofilm (25% weak, 56.2% moderate and 18.8% large amount), with significant difference between species (P < 0.001). Staphylococcus aureus and Staphylococcus epidermidis formed the largest amount of biofilm and had the largest number of CFU per ring. Staphylococci were most commonly resistant to penicillin (66.7%), tetracycline (50.0%), and erythromycin (45.8%); 41.7% of isolates was multidrug resistant and mecA gene was detected in five strains. All strains were susceptible to linezolid, vancomycin, teicoplanin and tigecycline.Staphylococci colonize LWs wedding rings, form biofilm on it, have multidrug resistant phenotype and/or carry mecA gene, representing a significant reservoir for the spreading of microorganisms and resistance. As far as we know, our study is the first that address this topic in laboratory workers.

Therapeutic potential of natural products and antibiotics against bovine mastitis pathogen of cows and buffaloes.

The present study aims to evaluate the prevalence and antimicrobial sensitivity of Staphylococcus aureus associated with bovine mastitis to selected antibiotics and plant extracts. In the current study, 140 milk samples were collected from cows and buffaloes. Among the 140 samples, 93 samples were positive for sub-clinical mastitis based on the California Mastitis Test (CMT). Out of the total positive samples, 45 were confirmed for S. aureus on a Mannitol salt agar media. The antimicrobial susceptibility test revealed that 44.82% of the isolates were resistant to cefoxitin (oxacillin) confirming methicillin-resistant S. aureus (MRSA) with a higher percentage (51.61%) in the buffalo than in the cow samples. Furthermore, the PCR assay confirmed the presence of the mecA gene in all the MRSA isolates. Among the seven tested antibiotics, sulfamethoxazole + trimethoprim showed high efficacy (71.1%) against methicillin-susceptible S. aureus isolates (MSSA). Oxytetracycline and sulfamethoxazole + trimethoprim showed 20% efficacy against MRSA followed by enrofloxacin (10%). On the other hand, the tested samples from Pistacia chinensis revealed that the ethyl acetate extract of bark showed a maximum zone of inhibition of 21.3 mm against MSSA and MRSA isolates at 3 000 µg/disc. Moreover, the methanol extract of Cotoneaster microphyllus formed a 12.3 mm and 9.1 mm zone of inhibition against the MSSA and MRSA isolates, respectively.

The NaHCO3-Responsive Phenotype in Methicillin-Resistant Staphylococcus aureus (MRSA) Is Influenced by mecA Genotype.

Methicillin-resistant Staphylococcus aureus (MRSA) strains are a leading cause of many invasive clinical syndromes, and pose treatment difficulties due to their in vitro resistance to most ß-lactams on standard laboratory testing. A novel phenotype frequently identified in MRSA strains, termed 'NaHCO3-responsiveness', is a property whereby strains are susceptible in vitro to many ß-lactams in the presence of NaHCO3. Specific mecA genotypes, repression of mecA/PBP2a expression and perturbed maturation of PBP2a by NaHCO3 have all been associated with this phenotype. The aim of this study was to define the relationship between specific mecA genotypes and PBP2a substitutions, on the one hand, with NaHCO3-responsiveness in vitro. Mutations were made in the mecA ribosomal binding site (RBS -7) and at amino acid position 246 of its coding region in parental strains MW2 (NaHCO3-responsive) and C36 (NaHCO3- nonresponsive) to generate 'swap' variants, each harboring the other's mecA-RBS/coding region genotypes. Successful swaps were confirmed by both sequencing, as well as predicted swap of in vitro penicillin-clavulanate susceptibility phenotypes. MW2 swap variants harboring the nonresponsive mecA genotypes became NaHCO3-nonresponsive (resistant to the ß-lactam, oxacillin [OXA]), in the presence of NaHCO3. Moreover, these swap variants had lost NaHCO3-mediated repression of mecA/PBP2a expression. In contrast, C36 swap variants harboring the NaHCO3-responsive mecA genotypes remained NaHCO3-nonresponsive phenotypically, and still exhibited nonrepressible mecA/PBP2a expression. These data demonstrate that in addition to the mecA genotype, NaHCO3-responsiveness may also depend on strain-specific genetic backgrounds.

Antimicrobial Susceptibility Testing for Staphylococcus lugdunensis.

Evaluation of penicillin and oxacillin susceptibility testing was conducted on 200 Staphylococcus lugdunensis isolates. Disc diffusion with penicillin 1 IU (P1, EUCAST) and penicillin 10 IU (P10, CLSI) was compared with nitrocefin discs (Cefinase) and automated broth microdilution (Vitek 2). Oxacillin susceptibility was extrapolated from cefoxitin (FOX; 30 µg) disc diffusion and compared with Vitek 2 results. The reference methods were blaZ and mecA PCR. Penicillin zone diameter and zone edge correlated with blaZ PCR results in all except two P10-susceptible isolates (very major error [VME]) and one P1-resistant isolate (major error [ME]). A total of 148 isolates were blaZ negative, of which 146 and 149 isolates were susceptible by P1 and P10, respectively. A total of 127 were penicillin susceptible by Vitek 2. Vitek 2 overcalled resistance in 21 blaZ-negative, 20 P1-susceptible, and 22 P10-susceptible isolates (Vitek 2 ME rate, 14.2%). Two mecA-positive isolates were oxacillin resistant by FOX disc and Vitek 2 methods (categorical agreement). However, 18 FOX-susceptible mecA-negative isolates tested resistant by Vitek 2. In conclusion, Vitek 2 overestimated penicillin and oxacillin resistance compared with disc diffusion and PCR results. In our study, disc diffusion with zone edge interpretation was more accurate and specific than automated broth microdilution for S. lugdunensis.

Study of antibiotics sensitivity pattern and molecular characterization of Staphylococcus aureus isolated from human and animal pyogenic cases.

Staphylococcus aureus has been described as the most common cause of human and animal diseases and has emerged as a 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 the isolates were subjected to PCR amplification to detect the existence of the Methicillin-resistant (mecA) and Coagulase (coa) genes. Lastly, typing was done 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 ß-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 five and six different band patterns in humans and animals, respectively, with two common patterns suggesting a common phylogenetic profile.

Distribution and Antibiotics Resistance Pattern of Community-Acquired Methicillin-Resistance Staphylococcus aureus in Southwestern Nigeria.

BACKGROUND: Methicillin-resistant Staphylococcus aureus is a global public health challenge and there is a continuous increase in community-acquired infections among people in different geographical location. We sought the distribution and antibiotics pattern of community-acquired methicillin-resistant Staphylococcus isolates among apparently healthy residents of Ibadan, Southwestern Nigeria. METHODS: Seven hundred (700) healthy volunteers residing in Ibadan metropolis, Nigeria, were enrolled in this study. Isolates from the nasal swabs were aseptically collected and characterized using standard and established microbiological methods, which included growth and fermentation on mannitol salt agar, colonial morphology, Gram-staining reaction, Microbact™ 12S identification kit and confirmed with 16SrRNA. After identification of the isolates, antimicrobial susceptibility test was performed on Mueller-Hinton agar by modified Kirby-Bauer disc diffusion method and the presence of mecA and nuc genes were detected via polymerase chain reaction assay. RESULTS: Prevalence of Staphylococcus aureus nasal carriage and Methicillin-resistant Staphylococcus in this study was 31.9% and 9.43% respectively. The residents of Ibadan North local government area (Fisher's Exact = 1.8962, P = .028) and Egbeda local government area (Fisher's Exact = 2.7222, P = .006) are likely to carry Methicillin-resistant Staphylococcus than any other local government area in Ibadan, Nigeria. The antimicrobial resistance patterns of the isolates revealed high resistance to Oxacillin (96.9%). Most of the isolates were sensitive to vancomycin (92.4%). Polymerase chain reaction analysis showed that mecA gene was present in all 66 (100%) Methicillin-resistant Staphylococcus aureus isolates. Male-gender (Ï°2 = 8.849, P = .003), Adults; 40-50 years old (Ï°2 = 9.842, P = .002), low educational background (Ï°2 = 36.817, P ˂ .001), recent hospital visitation (Ï°2 = 8.693, P = .003) are some of the factors that are observed in this study to be associated with Methicillin-resistant Staphylococcus infection. CONCLUSION: Our findings revealed the relatively high frequency of nasal carriers of Methicillin-resistant Staphylococcus aureus among the apparently healthy residents of the studied area and the advent of multidrug resistance among these isolates. Our study also supports previous findings on male-gender and low educational background as risk factors of S. aureus carriage. The need for rational chemotherapy, routine detection and regular surveillance of Methicillin-resistant Staphylococcus to limit its spread and reduce treatment failures is important.

Hand carriage, antimicrobial resistance and molecular characterisation of methicillin-resistant coagulase-negative staphylococci isolated from gynaecological surgical staff.

Sepsis caused by methicillin-resistant coagulase-negative staphylococci (MRCoNS) seriously affects the morbidity and mortality of neonates. However, the hand carriage and genotypic diversity of MRCoNS within surgical staff remain unclear in China. In the study, antimicrobial susceptibility tests and genotypic characterisation were applied to MRCoNS. One hundred and one samples were collected from the hands of gynaecological surgical staff. Eighty staphylococcal isolates were identified, of which 75 (94%) were resistant to at least one antibiotic. mecA gene was determined in 50 (62.5%) staphylococcal isolates. Panton-Valentine leukocidin (pvl) and ica genes were determined in 17 (21%) and 12 (15%) staphylococcal isolates, respectively. About 52% of staphylococci carried SCCmec IV and V, followed by SCCmec type I, II, and III (38%). In addition, two new STs types were assigned as ST844 and ST845. The high level of hand MRCoNS colonisation rate in gynaecological surgical staff is of concern, and hand hygiene management should be emphasised among surgical assistants.Impact statementWhat is already known on this subject? Coagulase-negative staphylococci (CoNS) are the predominant cause of neonatal sepsis. Exposure to antimicrobial-resistant CoNS might put neonates at increased risk of infection. However, little is known about the carriage and genetic diversity of methicillin-resistant CoNS (MRCoNS) of gynaecological surgeons and surgical assistants.What do the results of this study add? This is the first study on the molecular characterisation of MRCoNS recovered from gynaecological surgeons and surgical assistants in China. MRCoNS carriage rate in surgical assistants was significantly higher than in surgeons. Seventy-five (94%) coagulase-negative staphylococci were resistant to at least one antibiotic. SCCmec I, II and III was the dominant types identified in MRCoNS that were recovered from surgical staff. Fifty (62.5%) staphylococcal isolates that were recovered from surgical staff harboured the mecA gene. Pathogenic clones of MRCoNS were disseminated in surgical staff, and half of mecA-positive Staphylococcus epidermidis harboured the ica gene.What are the implications of these findings for clinical practice and/or further research? The high level of hand MRCoNS colonisation rate among gynaecological surgical staff is of concern. The alarming outcome of this study suggested that hygiene measures should be emphasised among gynaecological surgical assistants.

Methicillin resistant Staphylococcus aureus: A brief review of virulence and resistance.

Staphylococcus aureus is a common gram-positive human pathogen involved in both community-acquired and nosocomial infections ranging from localised superficial lesions to food poisoning and fatal systemic infections owing to its impressive array of virulence factors responsible for attaching, colonising, invading, and avoiding host immune system. The discovery of antibiotics effectively checked the once deadly infections. However, resistance started soon after their discovery and the first methicillin-resistant strain of staphylococcus aureus was reported in the early 1960s. The most important attribute of methicillin-resistant staphylococcus aureus is its acquisition of mecA gene coding for penicillin-binding protein-2a that blocks inhibitory action on peptidoglycan cross-linking. Methicillin-resistant staphylococcus aureus presents a serious global healthcare concern being responsible for prolonged hospital stays and increased mortality. The precise information of virulence factors and resistant traits of methicillin-resistant staphylococcus aureus and their interplay in a community is key to minimize the intermixing of resistant and susceptible pathogens in the community.

Phenotypic and genotypic characterization of oxacillin-susceptible and mecA positive Staphylococcus aureus strains isolated in Uruguay.

The aim of this study was to characterize phenotypically and genotypically 27 mecA positive Staphylococcus aureus strains with oxacillin MICs of ≤2µg/ml by Vitek 2, isolated in different regions of Uruguay. Susceptibility to oxacillin and cefoxitin was studied by gradient diffusion, disk diffusion to cefoxitin, and Phoenix and MicroScan systems. PBP2a was determined. SCCmec typing was performed and the isolates were compared by PFGE. Twenty-six isolates were susceptible to oxacillin; one strain was susceptible to cefoxitin by disk diffusion and 3 strains by gradient diffusion. Phoenix and MicroScan panels detected methicillin resistance in 25 and 27 strains, respectively. Twenty-six strains tested positive for PBP2a. Twenty-six strains carried SCCmec V and 24 belonged to pulsotype A. One strain carried SCCmec IV and did not belong to pulsotype A. Cefoxitin disk diffusion test and PBP2a detection correctly identified 26 of these 27 strains as MRSA. PFGE results suggest the dissemination of a cluster of MRSA carrying SCCmec V.

MOLECULAR EPIDEMIOLOGY OF THE TRANSMISSION OF METHICILLIN-RESISTANT STAPHYLOCOCCUS AUREUS IN KYIV ACUTE CARE HOSPITALS, UKRAINE.

OBJECTIVE: The aim: To evaluate the potential of transmission of methicillin-resistance Staphylococcus aureus (MRSA) in Ukrainian acute care hospitals. PATIENTS AND METHODS: Materials and methods: We performed a multicenter cross-sectional study. Definitions of HAI were used from the CDC/ NHSN. The susceptibility to antibiotics was determined by disk diffusion method according to the EUCAST. The cefoxitin-resistant isolates S.aureus were analyzed for the presence of the mecA gene and femA endogenous control gene using PCR. The virulence factor encoding genes (lukS-PV and lukF-PV) were detected by PCR. RESULTS: Results: Of 2,421 patients with HAIs caused S.aureus included in the study, 28.7% patients had MRSA. Prevalence of nasal carriage rate of MRSA among healthcare workers (HCWs) was 33.3%. MRSA contamination of hands and uniforms/gowns of HCW were 32.2% and 29.7%, respectively. MRSA contamination in the inanimate environment surfaces in near- and extended patients areas were 26.9%. The predominant MRSA contamination in hospital environment surfaces were: room inner door knob (32.8%), bed rails (28.9%), room light switch (28.9%), chair (27.9%), bedside table handle (20.6%), bedside table (20.5%), and tray table (13.7%). The PVL gene was present in 38.7% of MRSA strains, isolated from patients with HAIs and in 55.7% of MRSA, isolated from environment surfaces in patient area. In addition, the PVL genes were detected in over 56.3% of MRSA isolated from HCWs carrier. CONCLUSION: Conclusions: The majority of MRSA is acquired during hospitalization. Environmental surfaces may serve as potential reservoirs for nosocomial MRSA and facilitate transmissions via contact.

Phenotypic and Genomic Profiling of Staphylococcus argenteus in Canada and the United States and Recommendations for Clinical Result Reporting.

Staphylococcus argenteus is a newly described species, formerly known as S. aureus clonal complex 75 (CC75). Here, we describe the largest collection of S. argenteus isolates in North America, highlighting identification challenges. We present phenotypic and genomic characteristics and provide recommendations for clinical reporting. Between 2017 and 2019, 22 isolates of S. argenteus were received at 2 large reference laboratories for identification. Identification with routine methods (biochemical, matrix-assisted laser desorption ionization-time of flight mass spectrometry [MALDI-TOF MS], 16S rRNA gene analysis) proved challenging to confidently distinguish these isolates from S. aureus Whole-genome sequencing analysis was employed to confirm identifications. Using several different sequence-based analyses, all clinical isolates under investigation were confirmed to be S. argenteus with clear differentiation from S. aureus Seven of 22 isolates were recovered from sterile sites, 11 from nonsterile sites, and 4 from surveillance screens. While sequence types ST1223/coa type XV, ST2198/coa type XIV, and ST2793/coa type XId were identified among the Canadian isolates, the majority of isolates (73%) belonged to multilocus sequence types (MLST) ST2250/coa type XId and exhibited a high degree of homology at the genomic level. Despite this similarity, 5 spa types were identified among ST2250 isolates, demonstrating some diversity between strains. Several isolates carried mecA, as well as other resistance and virulence determinants (e.g., PVL, TSST-1) commonly associated with S. aureus Based on our findings, the growing body of literature on S. argenteus, the potential severity of infections, and possible confusion associated with reporting, including use of incorrect breakpoints for susceptibility results, we make recommendations for clinical laboratories regarding this organism.