Mannitol Salt Agar Research Articles

Comparative Study of Prevalence of MRSA on Surfaces of Different Laboratories in Bangalore in the COVID-19 Era

Background Methicillin-Resistant Staphylococcus aureus MRSA is endemic in India. MRSA prevalence ranges from 25 in the western part of India to 50 in the southern part. India is reporting an increase in community-acquired MRSA CA-MRSA. This research was carried out in six diagnostic centers for a period of one month during July 2020 to determine the prevalence of MRSA and susceptibility pattern of S. aureus isolates on high contact surfaces of NABL laboratories and non-NABL laboratories.Aim The goal of this study was to assess the performance of new infection control systems in the COVID-19 period in preventing MRSA colonization on high contact surfaces in laboratories and to see if they adhered to high standards NABL.Material amp Methods Total 96 swabs were taken from the following surfaces - waiting area collection room doorknob reception desk workbench incubator knob chair edges faucet knob. Two swabs were collected from each surface one was inoculated into nutrient agar and the other was inoculated on mannitol salt agar. The isolates were identified by appearance of colonies followed by catalase and coagulase tests and final identification was done by VITEK-2 system. Antibiotic sensitivity test was performed to check the MRSA strain.Results None of the surfaces of either NABL laboratories or non-NABL laboratories showed the growth of MRSA.Conclusion Based on the findings of the present study it can be concluded that all the procedures followed were highly effective for the control of the prevalence of MRSA by contact contamination.

Methicillin-Resistant Staphylococcus Auras (MRSA) from poultry meat products regarding mecA Gene, antibiotic Sensitivity, and biofilm Formation

The uprising problem of the Presence of Multi Drug Resistant (MDR) pathogenic bacteria is an emerging obstacle facing food safety. This study purposed to evaluate the antibiotic-resistant profile regarding the Presence of MRSA and biofilm production. About 225 samples were collected (60 imported frozen chicken meat, 30 chicken fillet, 30 chicken nuggets, 30 chicken shawarma, 15 chicken luncheon, and 60 chicken stock cubes) were examined bacteriologically for staphylococcus aureus using mannitol salt agar and blood agar, The occurrence of staphylococci was 127/225 (56.4%). 16.8% of the samples were S.aureus (38/225), while, 39.5% (89/225) were coagulase-negative staphylococci. S. aureus recovered from 8.3% of frozen chicken meat, 13.3% of chicken fillet, 23.3% of chicken nuggets, 46.6% of chicken shawarma, 33.3% of chicken luncheon, and 5% of chicken stock cubes. Resistance against 13 antibiotics was determined using Kirby-Bauer disc diffusion test, 100%, 78.9%, and 52.6% of the isolates were resistant to penicillin G, tetracycline, and amoxicillin. Clavulanic acid respectively, while 26.3% were resistant to methicillin. On the other hand, the isolates showed high sensitivity to vancomycin, clindamycin, and chloramphenicol with percentages 97.3%, 81.5%, and 68.4% respectively. The resistant strains to methicillin were tested by PCR for the presence of mecA gene, the result was 100% (10/10) positive for mecA gene. The isolates were phenotypically tested for the ability of biofilm formation using the Tube method that divided S. aureus isolates into 4 grades: strong biofilm former, moderate biofilm former, weak biofilm former, and non-biofilm former with percentages18.4%, 15.7%, 13.1%, and 52.6% respectively. To conclude, the increased prevalence of S.aureus in chicken meat products pinpoints the unhygienic condition of food processing, moreover, the occurrence of MDR S. aureus in chicken meat products might lead to alarming public health threats so there is an alerting need for rational use of antibiotics.

Multidrug-resistant Staphylococcus aureus isolated from chicken nuggets sold at superstores in Mymensingh, Bangladesh.

This study was conducted to determine the colony forming units (CFU) to isolate, identify, and antibiotic sensitivity of Staphylococcus aureus from chicken nuggets (CN). Sixty CN samples from two brands were collected from different superstores in Mymensingh, Bangladesh. Uncooked, oven-cooked (OC), or gas stove-cooked (GSC) CN samples were inoculated onto mannitol salt agar and blood agar. The total staphylococcal count (TSC) for uncooked CN ranged from log 4.68 to log 5.11 CFU/gm. For OC CN, TSC ranged from log 3.29 to log 3.62 CFU/gm. For GSC CN, TSC ranged from log 3.09 to log 3.49 CFU/gm. Relative to uncooked CN, microwave oven-cooked and GSC samples significantly reduced the TSC of CN (p < 0.01). Using the polymerase chain reaction assay and standard biochemical testing, only 8 out of 60 CN samples contained S. aureus. Staphylococcus aureus were resistant to Ampicillin (100%), Amoxicillin (100%), Oxacillin (75%), Cefixime (87.5%), Doxycycline (75%), intermediately sensitive to Erythromycin (25%), Cephalexin (12.5%), Ciprofloxacin (25%), Gentamicin (12.5%), Doxycycline (12.5%) and sensitive to Oxacillin (25%), Azithromycin (100%), Erythromycin (75%) Cephalexin (87.5%), Cefixime (12.5%), Chloramphenicol (100%), Ciprofloxacin (75%), Gentamicin (87.5%), Doxycycline (12.5%), and Vancomycin (100%). This study reports the first isolation and identification of S. aureus from CN in Bangladesh. GSC CN was better than OC and uncooked CN. Data also suggest that CN is contaminated with multidrug-resistant S. aureus, which poses a public health hazard.

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.

Nasal colonization of methicillin resistant staphylococcus aureus among slaughterhouse workers in Dar es salaam, Tanzania

Introduction: Methicillin resistant Staphylococcus aureus (MRSA) is a zoonotic pathogen that is associated with serious and sometimes fatal infections in humans. The pathogen has ability to acquire resistance to most antibiotics. Working in slaughterhouses increase the chance of workers to contract this pathogen. This study aimed to determine the prevalence of MRSA and its associated factors among slaughterhouse workers in Dar es Salaam, Tanzania. Methods: A cross-sectional study was conducted from June-July 2018 involving workers from 4 registered slaughterhouses. Social demographics and other information was collected using a semi-structured questionnaire. Swabs were collected and inoculated in Mannitol salt agar for S. aureus isolation. The isolates were tested for antimicrobial susceptibility using 10µg Oxacillin discs implanted on Muller Hinton Agar plates. Binary logistic regression was employed to determine the association between prevalence of MRSA and independent variables. Results: MRSA was isolated from 39,2% of the 258 enrolled slaughterhouse workers. MRSA colonization was more likely in persons who have been working for more than 21 years (p=0,014) and smokers (p=0,02). On-job training (adjOR=0,417; 95% CI: 0,202-0,858, p= 0,018) was protective against MRSA nasal colonization. Conclusion: Our study has demonstrated the occurrence of MRSA in slaughterhouse workers in Dar es salaam, Tanzania. It is more prevalent in persons who have been working for a long time in slaughterhouses. In addition, provision of on-job training is protective against MRSA nasal colonization. More studies are required to confirm whether the MRSA detected were livestock-associated.

A Preliminary Study on Antimicrobial Susceptibility of Staphylococcus spp. and Enterococcus spp. Grown on Mannitol Salt Agar in European Wild Boar (Sus scrofa) Hunted in Campania Region-Italy.

Simple SummaryDuring the last decade, an increase in the European wild boar (Sus scrofa) population occurred; thus, over the years, wild boars have become an important potential carrier of pathogenic bacteria for both livestock animals and pets, but also for humans. Since antibiotic resistance has become one of the greatest challenges of global public health, the aim of the present study was to define the prevalence and the antibiotic resistance profiles of bacteria grown on the selective medium mannitol salt agar (MSA), isolated from nasal swabs of wild boars hunted in Campania Region (southern Italy). The most prevalent isolated bacteria were represented by the Staphylococcus spp. and Enterococcus spp. strains, which showed worrying antibiotic-resistant profiles. Consequently, constant surveillance of wild boars is strongly recommended, in order to assess their role as reservoirs of antibiotic resistant bacteria and as sentinels of a possible environmental contamination.The importance of wild boar lies in its role as a bioindicator for the control of numerous zoonotic and non-zoonotic diseases, including antibiotic resistance. Mannitol Salt Agar (MSA) is a selective medium used for isolation, enumeration, and differentiation of pathogenic staphylococci. Other genera such as Enterococcus spp. are also salt tolerant and able to grow on MSA. The present study focused on the identification, by matrix assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS), of bacteria grown on MSA isolated from the nasal cavities of 50 healthy wild boars hunted in Campania Region (southern Italy) in the year 2019. In addition, the antimicrobial resistance phenotype of the isolated strains was determined by disk diffusion method. Among genus Staphylococcus, coagulase-negative Staphylococcus (CoNS) were the most common isolated species, with Staphylococcus xylosus as the most prevalent species (33.3%). Furthermore, Enterococcus spp. strains were isolated, and Enterococcus faecalis was the species showing the highest frequency of isolation (93.8%). For staphylococci, high levels of resistance to oxacillin (93.3%) were recorded. Differently, they exhibited low frequencies of resistance to tested non-β-lactams antibiotics. Among enterococci, the highest resistances were observed for penicillin (93.7%), followed by ampicillin (75%), and ciprofloxacin (68.7%). Interestingly, 43.7% of the isolated strains were vancomycin-resistant. In conclusion, this study reports the phenotypic antibiotic resistance profiles of Staphylococcus spp. and Enterococcus spp. strains isolated from nasal cavities of wild boars hunted in Campania Region, highlighting that these wild animals are carriers of antibiotic resistant bacteria.

Inhibition of the Growth of Staphylococcus Aureus by Using Some Antibiotics Produced from Bacillus Subtilis

Bacillus subtilis can produce many substances that are considered as a means of defense in addition to being tested and approved as antibiotics. The goal of this study was to see if Bacillus subtilis crude bacteriocin had any antibacterial effects on Staphylococcus aureus .The peptidic bacteriocin is considered of importance in the industrial and medical fields as an anticancer.We got our bacteria from the Bacterial Bank at the College of Science and the Department of Biology. Identification of the isolates was confirmed by microscopically and biochemical tests which include catalase, oxidase, IMViC, blood hemolysis, starch hydrolysis for Bacillus, and growth on Mannitol Salt Agar (MSA), Voges-Jenson agar(V.J), and Coagulase test for Staphylococcus. Based on the capacity of bacteriocin to diffuse in agar, the reverse technique, which eliminated the contact between the producers and sensitive strains, was used to detect bacteriocin antibacterial action. Results demonstrate the antibacterial effect of Bacillus subtilis bacteriocin by the inhibition of Staphylococcus aureus as compared to other indicator isolates.

Prevalence and Characterization of Methicillin-Resistant Staphylococcus aureus from Meat Retail Shops and Meat Handlers in the Buea Municipality, Cameroon

Aim: Methicillin-resistant Staphylococcus aureus (MRSA) is a notorious multidrug-resistant biovar of S. aureus associated with infections that result in high morbidity and mortality in humans and animals. Its persistence in both host and non-host environments adds a major ecological dimension to the problem, thereby making it a one health challenge. This study determined the prevalence and antibiotic resistance patterns of MRSA from meat retail shops and meat handlers in the Buea municipality in addition to basic hygiene practices implemented in the meat retail shops.&#x0D; Study Design: In this cross-sectional study, a total of 260 samples (comprising 52 swabs each from weighing balances, butchering slabs, cutting knives, hands of meat handlers and meat samples) were randomly obtained and subjected to microbiological analysis. Data on sociodemographic characteristics of meat handlers and basic hygiene practices were captured in a questionnaire.&#x0D; Methodology: After enrichment, each sample was inoculated on mannitol salt agar and DNA extracted from presumptive S. aureus isolates for detection of the nuc gene. MRSA was identified by oxacillin disk diffusion and confirmed by amplification of the mecA gene. All MRSA isolates were challenged with 10 commonly used antibiotics using the Kirby-Bauer disk diffusion method.&#x0D; Results: A total of 27 MRSA isolates were recovered from hand swabs (5.8%), meat (15.4%), knives swabs (21.2%) and butchering slabs (9.6%). All (100%) MRSA were resistant to ampicillin and cefepime followed by ofloxacin (92.6%), ciprofloxacin (81.5%) and vancomycin (37%). High susceptibility was observed for gentamicin (100%) and amikacin (100%). All isolates were multidrug-resistant and comprised 11 antibiotypes. Most meat handlers (71.2%) did not have basic meat safety training, while 90.4% of the meat retail shops did not have a nearby water source.&#x0D; Conclusion: Our results indicate that meat retail shops could be a potential reservoir of MRSA in the environment and meat handlers need more basic meat safety training.

Bacteriological Evaluation of Nigerian Paper Currency (Naira Notes) Circulating In Owerri, Imo State, Nigeria

Generally, the contamination of currencies with various microbial species is increasingly being reported. This usually results from improper handling during exchange of goods, services and certain environmental factors. This study on the bacteriological evaluation of the Nigerian paper currency (Naira notes) circulating in Owerri, Imo State was carried out with the aim of evaluating the prevalence of bacteria contaminants of Nigerian currency notes in circulation. A total of One hundred and twenty (120) Naira notes of ₦5, ₦10, ₦20, ₦50, ₦100, ₦200, ₦500 and ₦1000 denominations were collected in separate polythene bags from traders, students, hawkers, meat sellers, food vendors, taxi drivers, keke drivers and banks for the study. The notes were chosen on the basis of denominations and physical appearance (Mint, Neat, dirty, very dirty and mutilated). Each of the notes was inserted into a sterile bottle containing 10mls of distilled water and allowed to stand for twenty minutes. Double dilution of the solution was inoculated into Nutrient agar, MacConkey agar, Mannitol Salt agar and Salmonella and Shigella agar for viable counts. Further identification of the bacteria was carried out using standard morphological and biochemical tests. The data from this study were subjected to statistical analysis using percentage, charts and anova. The result from the analysis showed that, 82 (68.33%) out of the 120 samples evaluated were contaminated. The study showed that dirty naira notes are potential routes for bacteriological disease transmission to man during handling and constitutes a public health risk. Therefore, the appropriate authorities should embark on public enlightenment campaign targeted at the handlers and associated risks.

Dentistry and Intensive Care Unit: A Brief Report.

Objective The aim of this study is to verify whether removable dentures of patients admitted to an intensive care unit (ICU) are niches of microorganisms that can cause pathologies ( Staphylococcus aureus , Candida spp. , and enterobacteria). Materials and Methods Fifteen patients who were denture wearers (removable partial denture and complete denture) were included in this study. Patients must wear their dentures daily, and these dentures must have acrylic parts. Microbial biofilm was collected from the acrylic part of one denture of each patient. Then, the biofilm was seeded on different culture media: Sabouraud agar, blood agar, MacConkey agar, and mannitol salt agar. In this study, biochemical evaluations of microorganisms were performed. Statistical analysis The percentage of dentures with the microorganism identified by each culture medium was calculated. Results In total, 100% of the dentures were positive for Staphylococcus spp. (blood agar) and Candida spp. (Sabouraud agar); 33.3% of the dentures were positive for S. aureus (Mannitol salt agar); and 13.3% of the dentures were positive for Shigella spp. (MacConkey agar). Conclusion Removable dentures of patients (removable partial dentures and complete dentures) admitted to an ICU are niches of microorganisms that can cause pathologies.

Molecular and phylogenetic analysis of methicillin resistant Staphylococcus aureus isolated from subclinical mastitis in lactating ewes

This study was conducted for estimation of subclinical mastitis prevalence caused by Staphylococcus aureus (S.aureus) in apparent normal lactating ewes in the areas around Baghdad (Al-Rasheed, Al-Yousifya, Al-Latifya). 400 milk samples from 200 ewes were collected and examined by California mastitis test (CMT), Bacteriological examination was done by culturing on Blood agar, Mannitol salt agar, Staph-110 agar and Chrome agar then bacteria examined by Gram stain and biochemical test. Antibiotic sensitivity test and molecular assay by PCR were done to detect methicillin- resistant strains with the phylogenetic study. The results showed that 49/400 samples gave positive to CMT in the field and positive to S. aureus isolation. The percentage of subclinical mastitis in ewes was 12.25% in this study. All isolated bacteria were resistant to methicillin 100%. The result of PCR revealed that 16.32 % (8/49) of the MRSA were possess mecA gene, while no isolate carried the mecC gene, PCR-product for 8 positive samples of mecA target gene were sequenced, analyzed, and reported in Genbank-NCBI and obtained accession number and became a reference to Iraq and the Middle East and the world. Ongoing work will add to this set more type strains are published and it is available for download at ID: MZ359756.1, M Z359757.1, MZ359758.1, MZ359759.1, MZ359760.1, MZ359761.1, MZ359762.1, MZ359763.1 (mecA gene). Seven samples that match the global results by 100%, and one sample contains a mutation at the site 1539, Nucleotide. Our conclusion revealed that detection of subclinical mastitis in sheep very important caused by S. aureus and all S. aureus was resistant to methicillin and some strains possess mecA gene. However, absence of mec A in other MRSA isolates requires detecting the alternative genetic possibilities related to the resistance profile and fragment of gene used.

Occurrence of methicillin-resistant staphylococci in the pig-production chain in Ibadan, Nigeria.

Staphylococcus species colonises humans and animals and is a major food contaminant with public health significance. Here, we assessed the occurrence of methicillin-resistant staphylococci (MRS) in the pig-production chain in Ibadan, Nigeria. Nares of 120 pigs and 10 farmers were sampled with sterile swabs whilst 54 pork samples were collected from a retail slaughterhouse. Staphylococcus species were isolated using enrichment, cefoxitin–aztreonam selective broth and Mannitol salt agar. Isolates were tested for susceptibility to cefoxitin (30 μg), oxacillin (1 μg) and vancomycin (30 μg). Methicillin-resistant staphylococci isolates were characterised using conventional biochemical tests. From 184 samples, 364 staphylococcal isolates were obtained. Amongst the 54 pork samples, 44.0% were contaminated with Staphylococcus species. Overall, 9 (2.5%) MRS were obtained and presumptively identified as Staphylococcus xylosus (n = 3), Staphylococcus sciuri (n = 3), Staphylococcus warneri (n = 2) and Staphylococcus cohnii (n = 1). There was no relationship between the prevalence of MRS between pigs and pig handlers in the farms, but Farm 2 had the highest frequency of 66.7% (p < 0.05). Piglets had the highest prevalence of 66.7% (p < 0.05) whilst MRS was absent in workers and pork samples. This study raises concerns about the cross-contamination of staphylococci in the food chain. Constant surveillance is imperative to ensure food safety.

Bacterial factors of mastitis in lactating women and its effect on the physical properties and chemical composition of breast milk

Mastitis is a complication seen in some breastfeeding mothers and is the most common inflammatory lesion of the breast in breastfeeding mothers. In this complication, breast milk undergoes chemical and physical changes. It can lead to a drop in breastfeeding, weight loss, and, consequently, stunted growth of infants. Bacteria are the main cause of breast inflammation. Therefore, in this study, bacterial factors of mastitis were evaluated in lactating women. Also, their effects were considered on the physical properties and chemical composition of mothers' breast milk. For this purpose, 210 breastfeeding mothers referred to health centers were randomly selected, and their milk samples were collected. In addition to collecting mothers' demographic information by a questionnaire, the chemical composition (sugar, protein, and fat) and the physical properties (pH, density, and freezing temperature) of milk were measured. Bacterial evaluations were performed on the milk of these mothers by catalase test, coagulase test, and mannitol salt agar. Data were analyzed by SPSS software, Chi-square, Mann-Whitney U test, and T-test. The results showed that 56 mothers had mastitis, and Staphylococcus aureus and coagulase-negative staphylococci were the main bacteria in the milk of these mastitis mothers. These bacteria caused physical and chemical changes in breast milk so that mothers with Staphylococcus aureus mastitis had less sugar in their milk, and mothers with coagulase-negative staphylococci had less protein in their milk. Therefore, Staphylococcus aureus may reduce milk sugar by consuming milk sugar, and coagulase-negative staphylococci may also target milk protein. But to confirm these results, a larger population of mothers with mastitis is needed. Further studies are also needed to prove this result.

Prevalence of multidrug-resistant bacteria associated with polymicrobial infections

BackgroundWounds remain the most important cause of postoperative mortality and morbidity and generate considerable additional social and healthcare costs. Most wounds are caused by various coliforms, Enterococcus fecalis, Proteus sp., and multidrug resistant Staphylococcus aureus. Wound is one of the leading cause of infections in the under developed and developing countries than developed nations. MethodsA total of 43 samples associated with bacteremia and wound infection were collected. Biochemical characterization and culture characteristics of the drug resistant isolates were studied using MacConkey agar, blood agar and mannitol-salt agar. Antibiotic susceptibility analysis of the isolated strains was performed by disc diffusion method using various antibiotics. Prevalence of dug resistance among bacteria isolated from the wound was studied. The ability of Beta lactamase antibiotic producing bacterial strains were analyzed. ResultsA total of 168 bacterial strains were isolated showed high resistant towards ampicillin (89%), ciprofloxacin (90.8), cefepine (90.5), piperacillin (91.8), oxacillin (92.5), and imipenem (96.5). The isolated bacterial strains showed monobacterial as well as polybacterial growth on the surface of the wound. The isolated bacterial strains revealed 89% sensitivity against norfloxacin and 94.9 sensitivity against vancomycin. About 26% of bacterial strains degraded quinolones, whereas only 14% clinical isolates showed their ability to degrade aminoglycosides. A total of 27% bacteria degraded tetracycline and 51% of isolates degraded carbapenems compounds. Interestingly, E. faecalis was resistant against antibiotics such as, Oxacillin, Nalidic acid, Ofloxacin, Erythromycin, Norfloxacin, Ciprofloxacin, Ampicillin, Tetracycline, Cefepine, Amikacin, Cefurooxime, Vancomycin, Piperacillin, Imipenem and Gentamycin. Moreover, Proteus species was resistant against certain numbers of antibiotics namely, Ampicillin, Piperacillin, Oxacillin, Nalidic acid, Tetracycline, Erythromycin, Cefurooxime, Nitrofurantoin, Vancomycin and Imipenem. ConclusionsThe isolated bacterial strains were resistant against various drugs including vancomycin. Staphylococci, and E. faecalisis strains showed resistance against various classes of antibiotics.

Bacterial profile of high-touch surfaces, leftover drugs and antiseptics together with their antimicrobial susceptibility patterns at University of Gondar Comprehensive Specialized Hospital, Northwest Ethiopia

BackgroundThe hospital environment serves as a source of nosocomial infections, which pose a major therapeutic challenge. Although many bacteria species are common in hospital environments, their distribution, frequency, and antimicrobial susceptibility pattern from high-touch surfaces, leftover drugs, and antiseptics in different wards remain largely unknown. Hence, the aim of this study was to assess the magnitude and frequency of bacterial contaminants and their antimicrobial susceptibility patterns.MethodsA total of 384 samples were collected from five selected wards and processed according to standard bacteriological procedures. Samples were collected from high-touch surface using swabs and inoculated on Blood agar, MacConkey agar, Chocolate agar and Mannitol salt agar plates, and incubated at 37 °C for 24 h. On the other hand, the leftover drugs and 80% ethanol samples were collected using sterile cotton swab immersed in sterile tryptone soy broth then inoculated on culture medias and incubated at 37 °C for 24 h. Identification of bacteria species was done using the morphological characteristics, Gram stain, and biochemical tests while antimicrobial susceptibility tests were done using modified Kirby-Bauer disk diffusion technique following the Clinical Laboratory Standards Institute 2021guidelines.ResultsAmong the 384 samples processed, 102 (26.6%) were culture positive and a total of 114 bacterial isolates were identified. Gram-positive bacterial isolates were predominant, 64.9%, while Gram-negatives were 35.1%. The most frequently isolated bacteria were coagulase negative Staphylococci (38.6%) followed by S. aureus (13.2%) and P. aeruginosa (11.4%). On the other hand, the proportion of bacteria isolated from surgical ward, post-natal ward, orthopedic ward, trauma ward, and neonatal intensive care unit ward were 24.6, 21, 20.2, 18.4,15.8%, respectively. Sinks were mainly contaminated with Klebsiella species (81.8%) and A. baumannii (55.6%), while A. baumannii (22.2%) was the most contaminant for 80% ethanol. Gram-positive bacteria had significantly high resistance levels to penicillin (67.6%), cotrimoxazole (67.8%), and cefepime (80%). On the other hand, Gram-negative bacteria revealed the highest resistance levels to tetracycline (82.4%), amoxicillin-clavulanic acid (76.5%), cefepime (66.7%), ceftazidime (67.5%), and piperacillin (92.3%). Moreover, the proportion of multidrug resistant bacteria isolates was 44.7%.ConclusionsData of the present study showed that coagulase negative Staphylococci was the dominant bacterial isolates followed by S. aureus and P. aeruginosa. The proportion of multi-drug resistant bacteria isolates was relatively high. Therefore, appropriate infection prevention and control measures should be implemented.

Comparison of non-invasive Staphylococcus aureus sampling methods on lesional skin in patients with atopic dermatitis

There is evidence that Staphylococcus aureus colonisation is linked to severity of atopic dermatitis. As no gold standard for S. aureus sampling on atopic dermatitis skin lesions exists, this study compared three commonly used methods. In addition, effectiveness of standard skin disinfection to remove S. aureus colonisation from these inflamed skin lesions was investigated. In 30 atopic dermatitis patients, three different S. aureus sampling methods, i.e. detergent scrubbing, moist swabbing and tape stripping, were performed on naïve and disinfected skin lesions. Two different S. aureus selective media, mannitol salt agar and chromID agar, were used for bacterial growing. Quantifying the S. aureus load varied significantly between the different sampling methods on naïve skin lesions ranging from mean 51 to 1.5 × 104 CFU/cm2 (p < 0.001). The qualitative detection on naïve skin was highest with the two detergent-based techniques (86% each), while for tape stripping, this value was 67% (all on chromID agar). In comparison, mannitol salt agar was less sensitive (p < 0.001). The disinfection of the skin lesions led to a significant reduction of the S. aureus load (p < 0.05) but no complete eradication in the case of previously positive swab. The obtained data highlight the importance of the selected sampling method and consecutive S. aureus selection agar plates to implement further clinical studies for the effectiveness of topical anti-staphylococcal antibiotics. Other disinfection regimes should be considered in atopic dermatitis patients when complete de-colonisation of certain skin areas is required, e.g. for surgical procedures.

Methicillin-Resistant Coagulase Negative Staphylococci and Their Antibiotic Susceptibility Pattern from Healthy Dogs and Their Owners from Kathmandu Valley.

This cross-sectional study was designed to identify information on the frequency, antimicrobial resistance and species diversity of methicillin-resistant coagulase negative staphylococci (MRCoNS) among pet dogs and humans within households. Fifty five nasal swabs each from dogs and their owners were collected. MRCoNS were identified based on gram staining, culture on mannitol salt agar, biochemical tests, and mecA gene amplification. The antibiotic susceptibility of the isolates was assessed by a disc diffusion test. Uniplex and multiplex polymerase chain reaction (PCR) were employed for the species identification of MRCoNS and SCCmec typing, respectively. Species were further confirmed by MALDI-TOF-MS. The prevalence of MRCoNS was 29% in dog owners and 23.6% in dogs. Four different species of MRCoNS, Staphylococci saprophyticus (48.3%), S. haemolyticus (24.1%), S. warneri (17.2%), and S. epidermidis (10.3%), were detected. Two isolates each from dog owners and dogs showed a constitutive resistance to macrolide-lincosamide-streptogramin B (cMLSB) resistance, eight isolates each from dogs and their owners showed a macrolide-streptogramin B (MSB) resistance, and only two isolates from dog owners revealed an inducible resistance to macrolide-lincosamide-streptogramin B (iMLSB) resistance. SCCmec types were SCCmec type IV (55.2%), SCCmec type V (24.1%), SCCmec III (10.3%), SCCmec II (3.4%); two isolates were non-typable. MRCoNS are prevalent and genetically diverse in companion animals and humans. Different species of MRCoNS were found in dogs and their owners.

Antibiogram Susceptibility Patterns of Staphylococcus Aureus Harboring of MecA Gene and Prevalence Aminoglycoside Modifying Enzymes (AMEs) Genes in Iraq

Resistance to antibiotics, particularly aminoglycosides, has increased recently among Staphylococcus aureus strains, especially those resistant to methicillin and carrying the mecA gene. As a result, the purpose of this study was to examine the frequency of susceptibility patterns of aminoglycosides and predominance of genes regulated by aminoglycosides such as aac(6’)/aph(2”), aph(3”)-Ilia, and ant (4’)-Ia in MRSA isolates. In totality, there are 80 Staphylococcus aureus strains have been collected from a variety of clinical and environmental specimens and identified using standard culture and recognition requirements. MRSA strains were detected using cefoxitin disk diffusion and fermentation of mannitol salt agar methods, followed by molecular detection of the presence of the mec A gene. The agar disk diffusion technique was used to evaluate the antibiotic susceptibility of MRSA strains to aminoglycoside antibiotics. Multiplex PCR was used to test S. aureus for the existence of aminoglycoside-encoding genes, aac (6’)/aph(2”), aph(3”)-IIIa, and ant (4’)-Ia. From the total number of S. aureus isolates examined, 72 (90 %) were identified as MRSA, including every isolates contained the mec A gene. Susceptibility profile for MRSA isolates to aminoglycosides was as follows: gintamicin 12(16.6 %), amekacin 30(41.8 %), tobramycin 13(18.1 %), and netilmicin3 (4.1 %). The distribution of aac (6’)/aph(2”), aph(3”)-IIIa, and ant (4’)-Ia genes in MRSA isolates was 36.8 %, 0 %, and 0 %, respectively. Finally, because MRSA strains are a major problem in people’s diseases, This research’s findings might be valuable medical source providers in selecting appropriate medicines for the drug therapy of MRSA infections.

Antibacterial activity of Urena lobata against uropathogens

Urinary tract infections (UTI) are one of the most common form of bacterial infections but the treatment becomes cumbersome as the etiological bacteria are developing resistance against antibiotics. This present study evaluated the efficacy of antimicrobial activity of Urena lobata against uropathogens. Six urine samples from UTI patients were collected from Pathological Laboratory, G.B. Vaghani Multispecialty Hospital, Surat. Bacteria were isolated from these samples using Nutrient agar, Mac Conkey agar plate, Blood agar, Mannitol salt agar, Eosin Methylene Blue agar and King’s agar. The bacterial isolates were identified using cultural characteristics, microscopic features and biochemical characteristics. Leaf extract of Urena lobata was prepared using Soxhlet Extraction Method whereby methanol and distilled water were the extractants used. Herbal extract disc was prepared at concentrations of 50,75, and100 mg/ml and tested against all the isolates. DMSO and antibiotics (Nitrofurantion, Amikacin, Levofloxacin, Norofloxacin, Ofloxacin and Cephalosporins) were used as negative and positive controls respectively.Staphylococcus aureus, Escherichia coli, Bacillus cereus, Streptococcus pneumoniae, Klebsiella spp. and Brevibacillus panacihumi were isolated from the urine samples. All concentrations of aqueous and methanolic extracts of U. lobata leaf displayed highest zone of inhibition against B. cereus. No inhibitory effect was observed against the growth of Klebsiella except at the highest concentrations. Further study is encouraged on the in-vivo study of efficacy of U. lobata on etiological agent of UTI.

Determination of Microbial Pathogens and its Abundance in Fresh Kunun-Zaki Drinks: Huge Threat to Public Health

This study investigated the presence and abundance of microbial pathogens in fresh locally produced, packaged and distributed kunun-zaki drinks. A total of 20 samples were randomly purchased from 20 vendors and hawkers. Each of the samples obtained was tenfold serially diluted using sterile peptone water. From the appropriate dilutions, 0.1ml were removed and spread plated on Salmonella-Shigella agar, Mannitol salt agar, and Eosin methylene blue agar plates. The inoculated plates were then incubated at 37°C for 24 hours. This study found that of the 20 kunun-zaki samples investigated, 70% were contaminated by Staphylococcus aureus which ranged from 1.0x104 to 1.21x105 (CFU/ml). Salmonella spp ranged from 1.2x104 to 4.1x104 (CFU/ml), and contaminated 60% of the samples. Escherichia coli ranged from 0.2x104 to 1.1x104 (CFU/ml) and was found in 45% of the samples. The result showed that the locally produced, packaged and distributed kunun-zaki drinks were highly contaminated by foodborne microbial pathogens and it makes it unsafe for human consumption. It further showed that public health is put at risk when unsafe locally produced kunun-zaki beverage is consumed; hence, local producers and Agencies saddled with the responsibility to monitor, supervise and certify food safety must strictly ensure the protection of consumers and public health by insisting that traditional methods of production, packaging and distribution of kunun-zaki drinks follow, maintain and sustain standards that guarantee quality, safety and accessibility. Consumers must resist the urge to purchase kunun-zaki drinks from unverified sources; producers, vendors and hawkers.