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.

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

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.

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.

Evaluation of the Bacteriological Quality of Milk Sold in Nnewi, Nigeria

Introduction: Contamination of milk products can result to severe intestinal and extra-intestinal diseases in man. This study was aimed at evaluating the bacteriological quality of various milk products sold in Nnewi. Materials and Methods: Using the Cluster sampling technique, 30 milk samples (5 pasteurized skimmed milk, 5 powdered infant milk formulas, 5 powdered milk, 5 unsweetened evaporated milk, 5 branded soya milk and 5 unbranded soya milk) were purchased randomly from different shops and hawkers around Nnewi. Sample processing was done by serially diluting samples in sterile 1% peptone water before plating onto Mannitol Salt Agar, Violet Red Bile Glucose Agar, Blood Agar, MacConkey and Cysteine Lactose Electrolyte Deficient (CLED) agar using the Pour-plate technique. Bacterial count and identification were done using standard bacteriological as well as molecular techniques. The molecular techniques used were Polymerase Chain Reaction, Sanger Sequencing and BLAST analysis on the NCBI BLAST online. Results: This showed that 15 (50%) out of 30 milk samples were contaminated to varying degrees with bacteria. Nine (9) samples showed the presence of Escherichia coli with 32.14% of all the milk samples tested. E. coli was present in skimmed milk (20%), evaporated milk (20%), branded soyamilk (40%) and unbranded soyamilk (100%) but was not isolated from Infant formula and Powder milk. Klebsiella spp. showed the second highest prevalence (28.57%) and was present in evaporated milk (20%), branded soyamilk (40%), unbranded soyamilk (100%). Salmonella spp. (3.57%) was isolated from unbranded soyamilk,Enterobacter spp. (14.29%) was isolated from powder milk and in unbranded soyamilk samples, Staphylococcus aureus(3.57%) and Staphylococcus epidermidis (3.57%) were isolated only in unbranded soyamilk, Macrococcus caseolyticus (3.57%) was isolated from unbranded soyamilk. Novel species such as Aquitalea magnusonii (3.57%), Alishewanella fetalis (3.57%) and Lysinibacillus macroides (3.57%) were identified by molecular analysis to be present in infant formula, evaporated milk and unbranded soyamilk respectively. Conclusion: This research revealed that the bacteriological quality of some milk and milk products sold in Nnewi is not acceptable especially the unbranded soyamilk samples that showed gross contamination.

IDENTIFIKASI DAN UJI RESISTENSI Staphylococcus aureus TERHADAP ANTIBIOTIK (CHLORAMPHENICOL DAN CEFOTAXIME SODIUM) DARI PUS INFEKSI PIOGENIK DI PUSKESMAS PROPPO

Pemberian antibiotik pada pasien yang mengalami infeksi piogenik pada kulit di puskesmas proppo belum berdasarkan pada pola kuman dan sensitifitas antibiotik. Penelitian ini bertujuan untuk mengidentifikasi, isolasi serta uji sensitifitas bakteri terhadap beberapa antibiotik sehingga dapat diketahui jenis antibiotik yang paling paten untuk bakteri Staphylococcus aureus. Hasil pengamatan pada isolat pus menunjukkan adanya Staphylococcus aureus dengan ciri kesesuaian. Agar diperoleh bakteri murni dengan jumlah maksimum maka dilakukan isolasi. Telah berhasil diisolasi dan diidentifikasi ke-4 isolat pus yang tumbuh dan memfermentasi pada plat mannitol salt agar, sel berbentuk bulat bergerombol dan bersifat gram positif baik melalui pewarnaan maupun uji secara biokimia. Hasil uji terhadap Chloramphenicol memiliki nilai rata-rata daya hambat 24.0 mm dengan persentase penggunaan mencapai 75.0 %, sedangkan untuk Cefotaxime sodium memiliki nilai rata-rata zona hambat 10.9 mm dengan persentase penggunaan mencapai 25.0 %. Dapat disimpulkan bahwa ke-4 isolat tersebut kesemuanya diidentifikasi sebagai Staphylococcus aureus dengan zona hambat masing-masing isolat sebesar 23.5 mm, 24.2 mm, 24.0 mm dan 24.3 mm yang menunjukkan sensitif terhadap Chloramphenicol sedangkan zona hambat terhadap Cefotaxime sodium masing-masing isolat adalah 5 mm, 10.5 mm, 14.5 mm dan 13,5 mm yang menunjukkan resisten terhadap penggunaan antibiotik tersebut.

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.

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.

Elevators and staircase handrails as potential sources of nosocomial pathogens at Ndola Teaching Hospital, Zambia

This study aimed to assess microbial colonization of elevators and the staircase handrail at Ndola Teaching Hospital. Swabs from elevators and staircase handrail were cultured on Blood, MacConkey and Mannitol Salt agar for 24-48h at 35-37oC. All observed bacterial colonies were sub-cultured for identification. Data analysis was conducted with Microsoft excel 2010 and SPSS version 20 statistical software. A 2-tailed Pearson Correlation test was used to assess for significant differences in colonization prevalence between the two elevators and handrail. A total of 94 bacteria species were isolated, among which 75(78.8%) were isolated from elevators while 19(20.2%) were isolated from the staircase handrail. Most bacteria were isolated from exterior buttons followed by doors and interior of elevators whereas the basement, ground and sixth floor sections of the staircase handrail were largely contaminated with S. aureus, non-spore-forming Gram-positive bacilli, Klebsiella spp, coagulase-negative Staphylococci and Enterobacter spp. Overall, the commonest isolated bacteria were S. aureus (33%) followed by non-spore-forming Gram-positive bacilli (16%), coagulase-negative Staphylococci, and endospore-forming Gram-positive bacilli (13.8% apiece), Streptococci (7.4%), and Klebsiella species (6.4%). A strong relationship existed between the prevalence of bacteria colonizers of elevators and the staircase handrail (p<0.01). Therefore, the study showed that elevators and the staircase handrail possess viable microorganisms and may act as potential sources of nosocomial infections especially to immunocompromised patients in hospitals, and this calls for proper and effective infection control and prevention strategies to lessen microbial population from dry surfaces.

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

Background The 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. Methods A 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. Results Among 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%. Conclusions Data 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.