Reduced susceptibility to carbapenems in Enterobacteriaceae and antimicrobial resistance profile of Escherichia coli strains isolated from clinical and zoonotic sources in the Bamenda Municipality, Cameroon.

Introduction: food-producing animals harbour pathogenic and antibiotic resistant bacteria which can be transmitted to humans. Resistance to carbapenems may complicate treatment resulting to debilitating consequences. This study aimed at determining the susceptibility of Enterobacteriaceae to carbapenems and to compare the resistant patterns of E. coli strains isolated from clinical and zoonotic sources. Methods: this was a cross-sectional study involving patients presenting at the Bamenda Regional Hospital and abattoir samples. Clinical samples (faeces and urine) and zoonotic samples (cattle faeces) were cultured and isolates identified using API-20E. Enterobacteriaceae isolates were tested for their susceptibility to Carbapenems. The susceptibility of E. coli was tested against eight antibiotics on Mueller Hinton agar. Data was analysed using SPSS version 20. Results: Enterobacteriaceae isolates from clinical specimen showed susceptibility of 93.3% to carbapenems. Out of 208 isolates 14 (6.7%) were Carbapenem-resistant Enterobacteriaceae (CRE) while 30 (14.4%) showed intermediate resistance and 164 (78.9%) were susceptible. The predominant CRE were Proteus (7/16, 43.8%), Providencia (3/15, 20.0%) and E. coli (4/60, 6.7%) with E. coli being the most clinically significant CRE. Multiple drug resistance (MDR) was observed in 83% of E. coli isolates, with the highest resistance being against vancomycin (90, 81.8%), azithromycin (69, 62.7%) and doxycycline (68, 61.8%). Clinical isolates were significantly (P<0.05) more resistant to azithromycin, trimethoprim-suphamethoxazole and gentamicin than zoonotic isolates. Conclusion: CRE were detected among isolates and a high rate of multiple drug resistance was observed among E. coli isolates. Proper antibiotic policies and good hygiene/sanitation measures may curb the development/spread of CRE and MDR E. coli.

Zoonotic diseases risk perception and infection prevention and control practices among poultry farmers in the Buea Health District, Cameroon: A one health perspective.

Background and Aim: Livestock are associated with pathogenic microbes and farm workers play a significant role in the transmission of zoonotic diseases (ZDs). Lack of awareness of exposure risk among farmers may influence their farm practices, thereby enhancing the spread of diseases on farms and to the community. This study was aimed at evaluating the knowledge, risk perception, and prevention and control practices of ZDs among poultry farmers to provide baseline data for establishing a "One Health" practical approach to reducing ZD transmission in poultry farms. Materials and Methods: Using the exponential discriminative snowball technique, a community-based cross-sectional study involving poultry farmers was carried out in the Buea Health District from April to July 2021. Six feed-producing mills were used as focal points to identify and recruit farmers who were also referred to other farmers. Questionnaires were used to collect data related to participants' knowledge, risk perception, and prevention and control practices of ZDs. Descriptive analyses were performed for all variables while the chi-square test and logistic regression analysis were used to determine associations at 95% confidence level. Results: In all, 183 poultry farms and 207 workers were enrolled in the study. Despite being aware that animal diseases can be transmitted to humans, most participants showed poor knowledge (54.6%), low-risk perception (51.7%), and poor prevention/control practices (54.1%) on ZDs. The majority did not consider coming in contact with birds' body fluid (blood) or apparently healthy birds to be a risk of infection. More participants with small farms (<500 birds) had low-risk perception of ZDs than those with larger farms (>1000 birds) (p = 0.03). Furthermore, most participants reported practicing hand washing but they neither used protective devices such as gloves and face masks, and >50% would not invite veterinary professionals to their farms. There was a significant association between risk perception and knowledge (p = 0.007; CI = 1.257-4.200) as well as between risk perception and prevention/control practice (p = 0.002; CI = 1.451-4.867). Conclusion: Poultry farm workers in Buea had poor knowledge and perception of ZD risk and this might have contributed to their poor prevention/control practices on the farms. Enhanced informal education of poultry farmers through training workshops and seminars will improve their knowledge and skills on ZD transmission risk and prevention.

Pathogenic Potential and Antimicrobial Resistance Profile of Staphylococcus aureus in Milk and Beef from the Northwest and Southwest Regions of Cameroon.

Staphylococcus aureus is a major foodborne pathogen and commensal of the skin and mucous membranes of animals and humans. Its virulence relies on the production of a variety of toxins resistant to denaturing conditions. Increasing reports of S. aureus food poisoning and contamination of foods of animal origin elsewhere necessitates the investigation of these foods in Cameroon, to implement safety measures. This cross-sectional study evaluated S. aureus contamination in milk and beef in the Northwest and Southwest Regions of Cameroon, where cow milk is usually not pasteurized before consumption, and beef is the main source of protein. The distribution of antibiotic-resistant isolates and those with enterotoxin-producing potential was also investigated to provide data of public health and food safety benefit. S. aureus was isolated from 39 raw milk and 250 beef samples by standard methods. Confirmation of isolates was by PCR to detect the nuc gene. S. aureus was investigated for classical staphylococcal enterotoxin (SE) genes (sea, seb, sec, sed, and see) by PCR. Their susceptibility to 9 antibiotics was tested by the disk diffusion method. The chi-square test was used to compare the contamination of samples, antibiotic resistance, and the distribution of SE genes. S. aureus was isolated from 11.1% of samples. Contamination was higher in milk (48%) than in beef (5.2%) (P < 0.001). The sea was the most frequently (90%) harboured gene. A large proportion of isolates (88%) harboured more than one virulence gene. Isolates were generally resistant to erythromycin (82%), vancomycin (80%), tetracycline (76%), and oxacillin (74%). Multidrug resistance (MDR) was common (92%). Milk and beef samples in study area were contaminated with MDR enterotoxigenic S. aureus strains and may constitute a potential hazard to consumers. Thus, the need for implementation of proper hygienic measures when handling these products and pasteurization of milk cannot be overemphasized.

Occurrence of Antibiotic-Resistant Bacteria and Genes in Two Drinking Water Treatment and Distribution Systems in the North-West Province of South Africa.

With the increasing spread of antimicrobial resistance, there is growing attention to the contribution made by drinking water systems. The potential health impact of two drinking water treatment and distribution systems (A and B) in the North-West Province of South Africa was determined by investigating the water quality and occurrence of antimicrobial-resistant heterotrophic bacteria and genes in the raw and treated water over four seasons. Most of the physicochemical parameters except for electrical conductivity were within permissible limits. Coliform bacteria reduced from raw to potable water except for counts higher than the threshold recorded in Summer and Winter. A total of 203 heterotrophic bacterial isolates were recovered on chromogenic R2A medium and subjected to susceptibility testing to twelve antibiotics. Most of the isolates were resistant to ß-lactam antibiotics and Trimethoprim, whereas they were susceptible to Ciprofloxacin, Erythromycin, and Neomycin. The proportions of Cephalothin and Kanamycin-resistant isolates were significantly higher (p < 0.05) after treatment for site A, compared to significantly lower ß-lactam, Oxytetracycline, and Trimethoprim-resistant isolates for B. Over 50% of isolates were of high risk, indicating their origin from high antibiotic-use sources. Seventy-one (35%) isolates were multidrug-resistant, out of which the majority (53.5%, n = 38) possessed the strA gene, followed by strB 21 (29.6%), dfrB 13 (18.3%), aadA 11 (15.5%), blaCTX-M 5 (7.0%), and tetA 3 (4.2%). The 16S rRNA gene sequences of the isolates revealed strains belonging to eight bacterial families, some of which are clinically important.

Genotypic and Phenotypic Evaluation of Biofilm Production and Antimicrobial Resistance in Staphylococcus aureus Isolated from Milk, North West Province, South Africa.

Background: Biofilm formation in S. aureus may reduce the rate of penetration of antibiotics, thereby complicating treatment of infections caused by these bacteria. The aim of this study was to correlate biofilm-forming potentials, antimicrobial resistance, and genes in S. aureus isolates. Methods: A total of 64 milk samples were analysed, and 77 S. aureus were isolated. Results: Seventy (90.9%) isolates were biofilm producers. The ica biofilm-forming genes were detected among 75.3% of the isolates, with icaA being the most prevalent (49, 63.6%). The icaB gene was significantly (P = 0.027) higher in isolates with strong biofilm formation potentials. High resistance (60%-90%) of the isolates was observed against ceftriaxone, vancomycin, and penicillin, and 25 (32.5%) of S. aureus showed multidrug resistance (MDR) to at least three antibiotics. Five resistance genes, namely blaZ (29, 37.7%), vanC (29, 37.7%), tetK (24, 31.2%), tetL (21, 27.3%), and msrA/B (16, 20.8%) were detected. Most MDR phenotypes possessed at least one resistance gene alongside the biofilm genes. However, no distinct pattern was identified among the resistance and biofilm phenotypes. Conclusions: The high frequency of potentially pathogenic MDR S. aureus in milk samples intended for human consumption, demonstrates the public health relevance of this pathogen in the region.

Virulence profiles of vancomycin-resistant enterococci isolated from surface and ground water utilized by humans in the North West Province, South Africa: a public health perspective.

Vancomycin-resistant enterococci (VRE) have been responsible for numerous outbreaks of serious infections in humans worldwide. Enterococcus faecium and Enterococcus faecalis are the principal species that are frequently associated with vancomycin resistance determinants, thus usually implicated in hospital- and community-acquired infections in humans. The study aim was to determine the antibiotic resistance and virulence profiles of VREs isolated from surface and groundwater samples that are used by humans in the North West Province, South Africa. A total of 170 water samples were collected and analyzed. Eighty-one potential isolates were screened for characteristics of Enterococcus species using preliminary biochemical tests, PCR assays and sequence analysis. The antimicrobial resistance profiles of the isolates against nine antibiotics were determined and a dendrogram was generated to access the relatedness of the isolates. The isolates were screened for the presence of antibiotic resistance and virulence genes by multiplex PCR analysis. A total of 56 isolates were confirmed as Enterococcus species and the proportion of E. faecium (46.9%) was higher than E. faecalis (29%) and E. saccharolyticus (1.2%). Sequence data of E. faecium, E. faecalis, and E. saccharolyticus isolates revealed 97 to 98% similarities to clinical strains deposited in NCBI Genbank. Large proportions (44; 78.6%) of the isolates were resistant to vancomycin while 16 and 3.6% of the isolates possessed the vanA and vanB genes respectively. The MAR phenotype Vancomycin-Nalidixic Acid-Streptomycin-Chloramphenicol-Ampicillin-Oxytetracycline-Gentamycin-Nitrofurantoin-Sulphamethoxazole indicated that some isolates were resistant to all of the nine antibiotics tested. Cluster analysis of antibiotic resistance data revealed two major clusters. Sixteen (36.4%), 14 (27.3%), 3 (6.8%), and 2 (4.5%) of the VRE isolates possessed the gel, asa1, hyl, and esp virulence genes respectively while the cylA gene was not detected in the study. Multiple antibiotic-resistant enterococci were also resistant to vancomycin and possessed virulence determinants indicating that they can pose severe public health complications on individuals who consume contaminated water.