Coagulase-negative staphylococci from bovine milk: Antibiogram profiles and virulent gene detection.

BACKGROUND: Coagulase-negative Staphylococcus species are an emerging cause of intramammary infection, posing a significant economic and public health threat. The aim of this study was to assess the occurrence of coagulase-negative Staphylococcus species in bovine milk and dairy farms in Northwestern Ethiopia and to provide information about their antibiotic susceptibility and virulence gene profiles. METHODS: The cross-sectional study was conducted from February to August 2022. Coagulase-negative Staphylococcus species were isolated from 290 milk samples. Species isolation and identification were performed by plate culturing and biochemical tests and the antimicrobial susceptibility pattern of each isolate was determined by the Kirby-Bauer disc diffusion test. The single-plex PCR was used to detect the presence of virulent genes. The STATA software version 16 was used for data analysis. The prevalence, proportion of antimicrobial resistance and the number of virulent genes detected from coagulase-negative Staphylococcus species were analyzed using descriptive statistics. RESULTS: Coagulase-negative Staphylococcus species were isolated in 28.6%, (95% CI: 23.5-34.2) of the samples. Of these, the S. epidermidis, S. sciuri, S. warneri, S. haemolyticus, S. simulans, S. chromogens, S. cohnii, and S. captis species were isolated at the rates of 11, 5.2, 3.4, 3.1, 3.1, 1, 1, and 0.7% respectively. All the isolates showed a high percentage (100%) of resistance to Amoxicillin, Ampicillin, and Cefotetan and 37.5% of resistance to Oxacillin. The majority (54.2%) of coagulase-negative isolates also showed multidrug resistance. Coagulase-negative Staphylococcus species carried the icaD, pvl, mecA, hlb, sec, and hla virulent genes at the rates of 26.5%, 22.1%, 21.7%, 9.6%, 9.6% and 8.4% respectively. CONCLUSION: The present study revealed that the majority of the isolates (54.2%) were found multidrug-resistant and carriage of one or more virulent and enterotoxin genes responsible for intramammary and food poisoning infections. Thus, urgent disease control and prevention measures are warranted to reduce the deleterious impact of coagulase-negative species. To the best of our knowledge, this is the first study in Ethiopia to detect coagulase-negative Staphylococcus species with their associated virulent and food poisoning genes from bovine milk.

Antibiotic-resistant Enterobacteriaceae from diseased freshwater goldfish.

Goldfish farming gained more attention among the ornamental fishes in aquaculture industry. The occurrence of bacterial infections and further antimicrobial treatment lead to the major crisis of antibiotic resistance in aquaculture. We have isolated diverse enterobacteriaceae groups which affect the goldfish and identified their response towards 46 antimicrobials of 15 different classes. Thirteen significant bacterial isolates such as Edwardsiella tarda, Serratia marcescens, Klebsiella aerogenes, Proteus penneri, P. hauseri, Enterobacter cloacae, E. cancerogenus, E. ludwigii, Citrobacter freundii, E. coli, Kluyvera cryocrescens, Plesiomonas shigelloides and Providencia vermicola were recovered from the infected fish with the Shannon-wiener diversity index of 2.556. Multiple antibiotic resistance (MAR) index was found to be maximum for P. penneri (0.87) and minimum for C. freundii and E. cloacae (0.22), highlighting the hyper antibiotic selection pressure in the farm. The minimum concentration of antibiotics required to inhibit most of the resistant isolates was found to be > 256 mcg/ml. All the isolates were susceptible towards ciprofloxacin. Plasmid curing and further AMR tests could reveal the location of antibiotic resistance genes mainly as plasmids which determine the large extent of AMR spread through horizontal gene transfer. This study is the first of its kind to investigate the antimicrobial resistance profile of enterobacteriaceae recovered from goldfish, before and after plasmid curing.

Methicillin-resistant Staphylococcus aureus nasal carriage among janitors working in hospital and non-hospital areas: a comparative cross-sectional study.

BACKGROUND: Nasal colonization of Methicillin-resistant Staphylococcus aureus (MRSA) plays a key role in the epidemiology and pathogenesis of both healthcare-associated and community-acquired MRSA infections in various populations. Screening of MRSA nasal colonization is important in the prevention and control of infection and may provide useful information to guide antimicrobial therapy. This study aimed to determine nasal carriage of MRSA, its antimicrobial susceptibility pattern, and associated factors among janitors working in hospital & non-hospital areas at the University of Gondar, Northwest Ethiopia. METHODS: A comparative cross-sectional study was carried out in a total of 436 study participants (221 hospital and 215 non-hospital janitors) from January to May 2019. The study participants were sampled using a simple random sampling technique. Data on socio-demographic characteristics and associated factors were collected through face to face interviews using a structured questionnaire. Nasal swabs were collected and inoculated into Mannitol salt agar. MRSA was detected using cefoxitin (30 µg) disc and an antibiotic susceptibility test was done using the disc diffusion method. Data were entered and analyzed using SPSS version 20 statistical package. P value ≤ 0.05 was considered as statistically significant. RESULTS: The overall prevalence of S. aureus was 101/436 [23.2%, (95% CI: 19.3-27.8)], of which, 29.4% (65/221) were isolated from hospital and 16.7% (36/215) non-hospital janitors. The prevalence of MRSA was 4.8% (21/436) [95% CI: 3.0-6.9]; of these, 8.1% (18/221) of the isolates were from the hospital and 1.4% (3/215) non-hospital janitors, while methicillin-sensitive S. aureus (MSSA) in hospital & non-hospital janitors were 49 (22.2%) and 31 (14.4%), respectively. Among the MRSA isolates, 52.4% (11/21) were multi-drug resistant. Of these, 42.9% (9/18) were isolated from hospital and 66.7% (2/3) non-hospital janitors. Hence, nasal carriage of MRSA was significantly associated with hospitalization within the preceding year (AOR = 3.15, CI = 1.13-8.71). CONCLUSION: The present study revealed that high MSSA and MRSA were isolated from the hospital as compared to non-hospital janitors and high rates of antibiotics resistance were recorded in the hospital janitors. Consequently, hospitalizations were significantly associated with MRSA. Accordingly, regular screening of carriers in apparently healthy janitors is required for the prevention of nosocomial infections.

Occurrence, virulence gene and antimicrobial susceptibility profiles of Arcobacter sp. isolated from catla (Catla catla) in India.

In the present study, a total of 100 catla (Catla catla-major South Asian carp, local name botcha) collected from local fish markets and aquaculture ponds were subjected for isolation and characterization of Arcobacter sp. In all, 21 Arcobacter sp. were isolated, of which 18 (85·7%) were Arcobacter butzleri and three (14%) were A. cryoaerophilus as identified by multiplex PCR. All 18 A. butzleri isolates were positive for mviN, ciaB and tlyA virulence genes, three of A. cryoaerophilus isolates carried mviN gene and none of the isolates were positive for cadF, irgA, cj1349, hecA and hecB genes. All isolates (n = 21) were resistant to penicillin (100%). Meanwhile, 71·43, 23·81, 23·81, 14·29 and 9·52% of the isolates showed resistance towards vancomycin, nalidixic acid, erythromycin, cefixime and kanamycin, respectively. Multidrug resistance was observed in 23·81% of the Arcobacter sp. isolates and none of the isolates were positive for any of the extended spectrum beta-lactamases either by phenotypic or by molecular identification genes (blaOXA , blaSHV , blaTEM , blaCTX-M1 , blaCTX-M2 and blaCTX-M9 groups). The results emphasize the need to implement specific control procedures to reduce the use of antibiotics in aquaculture particularly the ones which are very important in human medicine. SIGNIFICANCE AND IMPACT OF THE STUDY: Arcobacter species are emerging food- and water-borne human pathogens. In this study, Arcobacter butzleri was predominant in fish compared to A. cryoaerohilus and A. skirrowii. Higher incidence of arcobacters in fish market samples suggests cross contamination and unhygienic handling of fish in markets. Virulence genes profile and antibiotics resistance of the Arcobacter sp. isolated in current study indicate pathogenic potential of Arcobacter sp. to humans. Occurrence of multidrug-resistant Arcobacter sp. in fish is a major concern in food safety. To our knowledge, this is the first report of Arcobacter sp. from freshwater fish, catla (Catla catla) in India.

Detection and antibiogram profile of diarrheagenic Escherichia coli isolated from two abattoir settings in northwest Ethiopia: a one health perspective.

BACKGROUND: Diarrheagenic Escherichia coli (E. coli) is a zoonotic pathogen that contaminates abattoir workers, slaughter environments, slaughter equipment, and carcasses during abattoir processing. Infection with E. coli is associated with the consumption of contaminated food and water, and it is a potential threat to the health and welfare of both humans and animals. Hence, this study aimed to detect diarrheagenic E. coli and assess its antibiogram profile in two abattoir settings, in one health lens. METHODS: A cross-sectional study in one health approach was conducted from December 2020 to June 2021. A total of 384 samples from abattoir workers' hands, carcasses, knives, cattle feces, abattoir water and effluents were collected. Bacterial culture and biochemical tests were conducted to isolate E. coli, while conventional polymerase chain reaction was performed to identify virulence genes. The antibiogram of diarrheagenic E. coli was tested against nine antimicrobials using the Kirby Bauer disk diffusion method. RESULTS: A total of 115 (29.95%) E. coli were isolated from the 384 samples, and from these isolates, about 17 (14.8%) were confirmed to be diarrheagenic E. coli (DEC). Among the DEC pathotypes, nine (52.94%), five (29.4%), and three (17.65%) were Shiga toxin-producing, enterohemorrhagic, and enterotoxigenic E. coli, respectively. While 14 (82.35%) DEC isolates harbored the stx2 gene, five (29.41%) the eae gene, five (29.41%) the hlyA gene and three (17.65%) harbored the st gene. All the DEC isolates were resistant to erythromycin and vancomycin; whereas, they were susceptible to ampicillin, nalidixic acid and norfloxacin. Furthermore, 64.7% of DEC isolates showed resistance to both ceftazidime and kanamycin and 88.24% of the isolates showed multidrug resistance. CONCLUSION: This study detected DEC isolates having different virulence genes, which showed single and multiple antimicrobial resistance. Given the existing poor hygienic and sanitary practices along the abattoir-to-table food chain, coupled with the habit of raw meat consumption, this result indicates a potential public and animal health risk from the pathogen and antimicrobial resistance.

Colibacillosis in lambs and kids in Egypt: Prevalence, serogroups, antibiogram profile, virulence genes distribution and antimicrobial resistance genes.

Background: Small ruminants have a socioeconomic impact on Egypt's production of meat, milk, and wool. Hence, every effort should be taken to prevent infections. Aim: To elucidate the prevalence and serogrouping of Escherichia coli (E. coli) strains from diarrheic lambs and kids, determine their antibiotic susceptibility and associated risk factors affecting the occurrence of the disease, and establish the most common virulence genes marker and major antimicrobial resistance genes. Methods: A total of 150 diarrheic animals (95 lambs and 55 kids) at different ages and seasons were subjected to clinical examination. Rectal swabs were collected from 150 diarrheic animals for isolation and biochemical identification of E. coli. Results: The bacteriological examination revealed that 62/95 lambs and 26/55 kids with percentages of 65% and 47%, respectively, showed infection with E. coli. Serotyping of 88 isolates of E. coli revealed the strains belonging to O2(8), O55(17), O84(5), O17(4), O6(8), O91(17), O26(9), O103(5), O126(5), O124(6), and O159(4). A total of 21 isolates were examined by multiplex polymerase chain reaction assay for detection of virulence and resistance genes. All examined isolates possessed a combination between intimin gene and heat-stable toxin (100%), the serine protease (pic) gene on 8/21 isolates of O55, O2, O6 (38%), and α-hemolysin gene on 8/21 isolates of O26, O91(38%) while adherent invasive gene (invA) gene on 3/21 isolates of O124, O159 (14%) which divided diarrheagenic E. coli into four types assigned to be atypical enteropathogenic E. coli (48%), atypical enterohemorrhagic E. coli 35%), atypical enterotoxigenic E. coli (6%), and atypical enteroinvasive E. coli (11%). On the other hand, the results of antimicrobial susceptibility testing revealed high resistance to ampicillin, erythromycin, and tetracycline (100%) and amoxicillin/clavulanic acid (92%) but were highly sensitive to gentamicin, imipenem, norfloxacin, ciprofloxacin, chloramphenicol, and amikacin (100%). Concerning to ß lactams antibiotic resistance genes of examined isolates had blaSHV (100%) and blaCTX-M (43%). For tetracycline, we detected the tetA in all examined isolates. Conclusion: The wide spread of atypical E. coli strains among diarrheic lambs and kids with marked resistance to several antibiotics of interest and the detection of major resistance genes assess the potential risk of this pathogen to animal and public health.

Microbiological profile and their antibiogram of bloodstream infections amongst first and second surge of the COVID-19 patients in a tertiary care hospital.

Introduction: The world is experiencing a pandemic of coronavirus disease 2019 (COVID-19) caused by SARS-CoV-2. The prescription of a superfluity of unnecessary antibiotics without regard for the potential for increased antimicrobial resistances is extensive and unimpeded during the COVID-19 pandemic. Aims: To compare the microorganisms and the pattern of antimicrobial resistance of bacteremia during the first and second waves of the COVID-19 pandemic in a tertiary care hospital. Methods and Material: This retrospective observational study, to compared the blood culture of the COVID-19 pandemic during the first wave (April 2020 to September 2020) and the second wave (April 2021 to September 2021). All the blood culture isolates were identified and the antimicrobial susceptibility testing was done according to standard guidelines. Results: Out of 1470 blood culture samples, 259 (17.6%) blood bacterial isolates were grown in the first wave and, out of 4200 blood culture samples, 711 (16.9%) bacterial isolated during the second wave of the COVID-19 pandemic. Coagulase-negative staphylococcus (CONS) was 32.8% followed by Staphylococcus aureus 29.7% in COVID first wave and staphylococcus aureus (48.9%) followed by Klebsiella pneumoniae (11.6%) during COVID second wave were the most prevalent isolates. Conclusions: This study shows that coagulase-negative staphylococcus aureus and multidrug-resistant Klebsiella spp. are the leading causes of bloodstream coagulase-negative infections during both the first and second wave in the bloodstream COVID-19 pandemic.

Isolation, identification, and antibiogram studies of Escherichia coli from ready-to-eat foods in Mymensingh, Bangladesh.

Background and Aim: Ready-to-eat (RTE) foods are widely used at home, restaurants, and during festivals in Bangladesh. So it is very important to investigate possible microbial contamination in RTE foods. Therefore, this study aimed to determine the total coliform count (TCC), isolate, identify, and characterize the Escherichia coli in RTE foods. The antimicrobial sensitivity of E. coli obtained from RTE foods was also performed using 12 commonly used antibiotics. Materials and Methods: A total of 100 RTE food samples were collected aseptically and comprised of ten samples each: Burger, pizza, sandwich, chicken roll, chicken meat loaf, chicken fry, salad vegetable, ice-cream, yogurt, and milkshake sold in Mymensingh City. Samples were inoculated onto Eosin methylene blue agar and incubated at 37°C for 24 h. Isolation and identification of bacteria were performed based on cultural, staining, and biochemical properties, followed by a polymerase chain reaction. Results: The TCC in Chicken meat loaf, burger, pizza, sandwich, salad vegetable ice-cream, and yogurt samples were 3.57 ± 0.96, 3.69 ± 0.08, 3.50 ± 0.60, 2.60 ± 0.20, 4.09 ± 0.29, 4.44 ± 0.25, and 3.14 ± 0.30 mean log colony-forming units ± standard deviation/mL, respectively. The study found a higher prevalence of E. coli in RTE salad vegetable products than in RTE meat and milk products. Forty percent of the mixed vegetable salad samples showed positive results for E. coli. Whereas E. coli prevalence in RTE meat and milk products was 20% and 16.7%, respectively. All the 21 isolates were subjected to antibiotic susceptibility test against 12 different antibiotics. It was observed that 46.1% were susceptible, 16.6% were intermediate, 46.1% were resistant, and 47.6% were multidrug-resistant (MDR) among seven different antibiotic classes. E. coli isolates were resistant to cephalexin, ceftazidime, oxytetracycline, and ampicillin and sensitive to gentamycin, followed by kanamycin, ceftriaxone, colistin, and enrofloxacin. Conclusion: The study revealed that RTE foods are a serious issue from a public health point of view. To achieve a safer level of E. coli in RTE foods sold for human consumption, public food outlets must improve hygienic and good production procedures. Moreover, MDR E. coli in these foods pose serious public health threats.

Epidemiology and Antibiogram Profile of Vibrio cholerae Isolates between 2004-2013 from Odisha, India.

Cholera is an acute diarrheal disease caused by Vibrio cholerae serogroups O1 and O139, which are known to cause epidemics of cholera in Odisha. The present study was intended to document the antibiotic resistance pattern among clinical isolates of both serogroups of V. cholerae (O1 and O139) isolated during 2004-2013. Nine-hundred nine isolates of V. cholerae were included in this study and were identified by standard procedures. An antibiotic sensitivity test was performed by the disc diffusion method. The seasonality of cholera in this region indicated that there was one peak in the rainy season only. The number of cholera cases started increasing from July and declined starting from the month of October onward. The adult age group of patients was the worst affected among all age groups of patients. The 2 different serogroups of V. cholerae (O1 and O139) showed different prevalence rates (%) of resistance to all the antibiotics in each year. Serogroup O1 showed uniformly high resistance to co-trimoxazole, furazolidone, and nalidixic acid throughout the study. Chloramphenicol encountered resistance only during 2009, but the strains were sensitive in the other years. The emergence of multiple drug-resistant V. cholerae strains may significantly influence the control of future outbreaks and epidemics of cholera in this region.

Isolation and identification of bacteria from fresh guava (Psidium guajava) sold at local markets in Mymensingh and their antibiogram profile.

AIM: The study was conducted for the isolation, identification, and antibiogram of bacteria obtained from fresh guava (Psidium guajava). MATERIALS AND METHODS: A total of 25 fresh guavas were collected from five markets located in Mymensingh city. Guava samples were cultured onto various selective media such as eosin methylene blue, xylose lysine deoxycholate, thiosulfate-citrate-bile salts-sucrose, blood agar, and mannitol salt agar for the isolation of bacteria. Biochemical tests (dextrose, maltose, lactose, sucrose, mannitol, methyl red, Voges-Proskauer, and indole) were performed to identify the bacteria. RESULTS: Total viable counts of guava were ranged between log 6.56 colony-forming unit (cfu)/ml and 6.62 cfu/ml. A total of 106 bacterial isolates belonged to five genera (Escherichia coli, Salmonella spp., Vibrio spp., Bacillus spp., and Staphylococcus spp.) were identified. Salmonella spp. (23.6%) was the most prevalent, followed by E. coli (22.64%), Bacillus spp. (19.81%), Staphylococcus spp. (17.92%), and Vibrio spp. (16.03%). The results of antibiotic sensitivity test showed that Salmonella spp., Bacillus spp., and E. coli were sensitive to chloramphenicol, ciprofloxacin, and gentamicin and resistant to ampicillin and cephalexin. Vibrio spp. was sensitive to chloramphenicol and gentamicin, intermediately sensitive to ciprofloxacin and ampicillin and resistant to cephalexin. CONCLUSION: The results of this study indicate that fresh guava contains multidrug-resistant bacteria which might pose a public health risk.