Prevalence of non-O157 Shiga Toxin-producing E. Coli in Children and Calves in Al- Muthanna Province, Iraq

The present study focus on non-O157 Shiga toxin-producing E. Coli (STEC), included a bacteriological study was subjected to provide additional information for non-O157 STEC prevalence in children and calves. Isolation by using selective culturing media (CHROMagar STEC and CHROMagar O157) from 127 children suffering from diarrhea and 133 calves in Al- Muthanna province. Characterization depends on culturing positive colony on MacConkey agar and Levin’s Eosin Methylene blue agar, staining single colony from the growth by gram stain, biochemical tests; Indole, the Methyl Red, Voges-Proskauer, Citrate test, Oxidase, Catalase, Urease, Motility, Kligler Iron and Api-20E, were done to confirm a diagnosis of non-O157 STEC, The reliable isolation as non-O157 STEC serotyping by specific latex agglutination test for the target non-O157 STEC (big six) serogroup (O26, O45, O103, O111, O121 and O145). The current study showed the prevalence of non-O157 STEC was 20 of out 127 (15.73%) in samples collected from children and 27 / 133 (20.30%) in calves samples in conclusion the Non-O157 STEC is an important cause of diarrhea in children, and calves; finally, the calves play an important reservoir for Non-O157 STEC.

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.

Profile variation of bla genes among non-lactose fermenting Gram-negative bacilli between clinical and environmental isolates of Dr. Soetomo Hospital, Surabaya, Indonesia

Endraputra PN, Kuntaman K, Wasito EB, Shirakawa T, Raharjo D, Setyarini W. 2021. Profile variation of bla genes among non-lactose fermenting Gram negative bacilli between clinical and environmental isolates of Dr. Soetomo Hospital, Surabaya, Indonesia. Biodiversitas 22: 5047-5054. Carbapenem-resistant non-fermenter Gram-negative bacilli are notorious opportunistic pathogens in hospitalized patients and hospital environments. This study explored the carbapenemase gene among non-fermenter Gram-negative bacilli from hospital wastewater and clinical isolates in Dr. Soetomo Hospital, Surabaya, Indonesia. All samples were screened on MacConkey agar with meropenem 2 µg/ml and gene detected by Multiplex PCR. All samples were screened on MacConkey agar with meropenem 2 µg/ml and gene detected by Multiplex PCR. A total of 121 isolates consisted of 76 clinical (41 carbapenem-resistant Acinetobacter baumannii and 35 carbapenem-resistant Pseudomonas aeruginosa), 45 environmental isolates (6 carbapenem-resistant Pseudomonas aeruginosa and 32 carbapenem-resistant Pseudomonas spp.), and 7 screening samples (all CRPAs). Clinical isolates carbapenemase genes were identified, blaOXA-23-like 21 (28%), blaOXA-24-like 30 (39%), blaNDM-1 1 (1%), and blaIMP-1 6 (8%) while environmental isolates were blaOXA-23-like 5 (13%), blaOXA-24-like 4 (11%), blaOXA-48-like 2 (5%), blaNDM-1 13 (34%), blaVIM 12 (32%), and blaIMP-1 4 (11%). Rectal swab screening specimens presented blaOXA-23-like 3 (43%), blaOXA-24-like 3 (43%), and blaNDM-1 1 (14%). The carbapenemase gene pattern was different between clinical and environmental isolates. The blaOXA-23-like and blaOXA-24-like were most prevalent among in both clinical and wastewater, while blaVIM was mostly in wastewater. The presence of carbapenem-resistant non-fermenter Gram-negative bacilli carrying carbapenemase genes in hospital effluents indicated that the community river was seeded with an antimicrobial resistance gene.

High level of intrinsic phenotypic antimicrobial resistance in enterobacteria from terrestrial wildlife in Gabonese national parks

Data on the prevalence of antibiotic resistance in Enterobacteriaceae in African wildlife are still relatively limited. The aim of this study was to estimate the prevalence of phenotypic intrinsic and acquired antimicrobial resistance of enterobacteria from several species of terrestrial wild mammals in national parks of Gabon. Colony culture and isolation were done using MacConkey agar. Isolates were identified using the VITEK 2 and MALDI-TOF methods. Antibiotic susceptibility was analysed and interpreted according to the European Committee on Antimicrobial Susceptibility Testing guidelines. The preliminary test for ESBL-producing Enterobacteriaceae was performed by replicating enterobacterial colonies on MacConkey agar supplemented with 2 mg/L cefotaxime (MCA+CTX). Extended-spectrum beta-lactamase (ESBL) production was confirmed with the double-disc synergy test (DDST). The inhibition zone diameters were read with SirScan. Among the 130 bacterial colonies isolated from 125 fecal samples, 90 enterobacterial isolates were identified. Escherichia coli (61%) was the most prevalent, followed by Enterobacter cloacae (8%), Proteus mirabilis (8%), Klebsiella variicola (7%), Klebsiella aerogenes (7%), Klebsiella oxytoca (4%), Citrobacter freundii (3%), Klebsiella pneumoniae (1%) and Serratia marcescens (1%). Acquired resistance was carried by E. coli (11% of all E. coli isolates) and E. cloacae (3% of all E. cloacae) isolates, while intrinsic resistance was detected in all the other resistant isolates (n = 31); K. variicola, K. oxytoca, K. pneumoniae, E. cloacae, K. aerogenes, S. marcescens and P. mirabilis). Our data show that most strains isolated in protected areas in Gabon are wild type isolates and carry intrinsic resistance rather than acquired resistance.

Detection of Escherichia albertii in retail oysters

Escherichia albertii is an emerging foodborne pathogen. Owing to its distribution in river water, it is important to determine the presence of E. albertii in aquaculture-related foods. In this study, we investigated the distribution of E. albertii in retail oyster samples. A total of 427 raw oyster samples (385 Pacific oysters, and 42 Japanese rock oysters) were enriched in modified Escherichia coli broth (mEC) or mEC supplemented with novobiocin (NmEC) at 42 °C. The cultures were used for E. albertii -specific nested PCR assay, as well as for E. albertii isolation using deoxycholate hydrogen sulfide lactose agar (DHL), DHL supplemented with rhamnose and xylose (RX-DHL), and MacConkey agar supplemented with rhamnose and xylose (RX-MAC). The population of E. albertii in nested PCR-positive samples was determined using the most probable number (MPN) method. E. albertii isolates were subjected to biochemical and genetic characterization. E. albertii was detected in 5 of 315 (1.6%) Pacific oyster samples (one piece each), 2 of 70 (2.9 %) Pacific oyster samples (25 g each), and 2 of 42 (4.8 %) Japanese rock oyster samples procured from four geographically distant regions. A total of 64 E. albertii strains were isolated from eight of the nine nested PCR assay-positive oyster samples, and the MPN value was under the detection limit (< 3 MPN/10 g). A specific season or month for detecting E. albertii was not observed in this study, suggesting that the pathogen is present in seawater. All the E. albertii isolates, except one, were positive for the virulence factor eae, indicating that these isolates have the potential to infect humans.

Bacteria variety causing clinical mastitis in Holstein-Friesian cows in Pelagonia region, North Macedonia

The main aim of this study are the bacteria that most often cause clinical mastitis (CM) and their impact on milk reduction in Holstein-Friesian cows in the Pelagonia – North Macedonia region. 36 milk samples were taken from Holstein-Friesian breed of cows with confirmed clinical mastitis by a veterinarian. The samples were taken for the period from January 2019 to December 2020 from 20 different smallholder farms situated in the monitored region. Two sterile tubes with 10 ml of milk in each of them were taken from the affected part of the udder of the cow. A total of 86 tubes with milk from 36 mastitis cows were taken. From each sample 300 µl drips were placed in petri dishes with different selective nutrient media: Mannitol Salt Agar, MacConkey Agar, Endo Agar and Edwards nutrient medium. The petri dishes were incubated at 35±2°C for 24-48 hours in Mannitol Salt Agar, at 30-35°C for 18 to 72 hours in MacConkey Agar, at 35±2°C for 18 to 24 hours in Endo Agar and at 35-37°C for 24-48 hours in Edwards nutrient medium. Morphology of colonies and cells were examined with a microscope. A total of 119 strains were obtained and the following physiological and biochemical studies were performed to determine the new isolates: oxidase reaction, catalysis activity, indol test, hydrolysis of the hyporate, acetoin formation (acetylmethylcarbinol, Voges-Proscauer reaction) and Methyl-Roth test (MR- test). The results obtained revealed that the most common bacterial species causing clinical mastitis in Holstein-Friesian cows in 2019 and 2020 were six species of bacteria, where E. coli and Staphylococcus spp. are dominant.

A SEARCH FOR ASSOCIATION BETWEEN BIOFILM STRENGTH AND SITES OF INFECTION CAUSED BY ACINETOBACTER BAUMANNII COMPLEX

INTRODUCTION: In the world of infectious diseases, a group of bacteria known as ESKAPE pathogens, are responsible for most of the life threatening multidrug resistant (MDR) and extensively drug resistant (XDR) infections worldwide. The ESKAPE pathogens comprise of Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, Enterobacter spp. AIMS AND OBJECTIVES: The objective of this study is isolate and identify Acinetobactor baumannii complex from clinical samples. Measure the biolm forming capacity of the isolates. Possible association of this biolm strength with the type of clinical sample from which they are isolated. MATERIALS AND METHODS: The samples which were tested in the laboratory were the following: Sputum, End tracheal tube aspirates, Pus, Urine, Blood for culture, Central venous catheter tips & Cerebrospinal uid. Following culture media were taken: MaCconkey agar & Blood agar. The colonies suggestive of Acinetobacter baumannii were selected. On MaCconkey agar, non- pigmented NLF colonies with pink to light lavender hue; and on blood agar, opaque non-hemolytic colonies were obtained. Gram stains from selected colony showed gram negative coccobacilli. Hanging drop preparation showed non-motile bacteria. RESULTS AND ANALYSIS: We found maximum number of strong biolm producers were found among the isolates causing infection of CVC tips (70%). This was closely followed by ETtube isolates (69.5%). The isolates from blood stream, urine, and sputum samples showed all the three patterns, namely, no biolm producer, moderate biolm producer, and strong biolm producer. Our study showed interestingly, no strong biolm producer was found from pus samples, whereas from the CSF isolates no non-producer of biolm emerged. CONCLUSION: This study centers around strength or pattern of biolm formation by this bacteria. It was interesting to nd that there was a statistically signicant association between patterns of biolm produced by Acinetobacter baumannii and the various samples from which they are isolated. This made us to hint at a probable relation between biolm patterns and organotropism as an hypothesis. Only further investigations will tell if it can stand the test of time.

Validation of selective agars for detection and quantification of Escherichia coli resistant to critically important antimicrobials

Success in the global fight against antimicrobial resistance (AMR) is likely to improve if surveillance can be performed more rapidly, affordably and on a larger scale. An approach based on robotics and agars incorporated with antimicrobials has enormous potential to achieve this. However, there is a need to identify the combinations of selective agars and key antimicrobials yielding the most accurate counts of susceptible and resistant organisms. A series of designed experiments involving 1,202 plates identified the best candidate-combinations from six commercially available agars and five antimicrobials using 18 Escherichia coli strains as either pure cultures or inoculums within faeces. The effect of various design factors on colony counts were analysed in generalised linear models. Without antimicrobials, Brilliance™ E. coli (Brilliance) and CHROMagar™ ECC (CHROMagar) agars yielded 28.9% and 23.5% more colonies than MacConkey agar. The order of superiority of agars remained unchanged when faecal samples with and without spiking of resistant E. coli were inoculated onto agars with or without specific antimicrobials. When incorporating antimicrobials at varying concentrations, it was revealed that ampicillin, tetracycline and ciprofloxacin are suitable for incorporation into Brilliance and CHROMagar agars at all defined concentrations. Gentamicin was only suitable for incorporation at 8 and 16 µg/mL while ceftiofur was only suitable at 1 µg/mL. CHROMagar™ ESBL agar supported growth of a wider diversity of extended-spectrum cephalosporin-resistant E. coli. The findings demonstrate the potential for combining robotics with agars to deliver AMR surveillance on a vast scale with greater sensitivity of detection and strategic relevance.

Isolation Procedure for CP E. coli from Caeca Samples under Review Towards an Increased Sensitivity

Due to the increasing reports of carbapenemase-producing Enterobacteriaceae (CPE) from livestock in recent years, the European Reference Laboratory for Antimicrobial Resistances (EURL-AR) provided a protocol for their recovery from caecum and meat samples. This procedure exhibited limitations for the detection of CPE with low carbapenem MIC values. Therefore, it was modified by a second, selective enrichment in lysogeny broth with cefotaxime (CTX 1 mg/L) and with meropenem (MEM 0.125 mg/L) at 37 °C under microaerophilic conditions. By Real-time PCR, these enrichments are pre-screened for the most common carbapenemase genes. Another adaptation was the use of in-house prepared MacConkey agar with MEM and MEM+CTX instead of commercial selective agar. According to the EURL-method, we achieved 100% sensitivity and specificity using the in-house media instead of commercial agar, which decreased the sensitivity to ~75%. Comparing the method with and without the second enrichment, no substantial influence on sensitivity and specificity was detected. Nevertheless, this enrichment has simplified the CPE-isolation regarding the accompanying microbiota and the separation of putative colonies. In conclusion, the sensitivity of the method can be increased with slight modifications.