Recent Developments in Phenotypic and Molecular Diagnostic Methods for Antimicrobial Resistance Detection in Staphylococcus aureus: A Narrative Review

Staphylococcus aureus is an opportunistic pathogen responsible for a wide range of infections in humans, such as skin and soft tissue infections, pneumonia, food poisoning or sepsis. Historically, S. aureus was able to rapidly adapt to anti-staphylococcal antibiotics and become resistant to several classes of antibiotics. Today, methicillin-resistant S. aureus (MRSA) is a multidrug-resistant pathogen and is one of the most common bacteria responsible for hospital-acquired infections and outbreaks, in community settings as well. The rapid and accurate diagnosis of antimicrobial resistance in S. aureus is crucial to the early initiation of directed antibiotic therapy and to improve clinical outcomes for patients. In this narrative review, I provide an overview of recent phenotypic and molecular diagnostic methods for antimicrobial resistance detection in S. aureus, with a particular focus on MRSA detection. I consider methods for resistance detection in both clinical samples and isolated S. aureus cultures, along with a brief discussion of the advantages and the challenges of implementing such methods in routine diagnostics.

Campylobacter jejuni Virulence Factors Identified by Modulating Their Synthesis on Ribosomes With Altered rRNA Methylation

Campylobacter jejuni is a major cause of food poisoning worldwide, and remains the main infective agent in gastroenteritis and related intestinal disorders in Europe and the USA. As with all bacterial infections, the stages of adhesion to host tissue, survival in the host and eliciting disease all require the synthesis of proteinaceous virulence factors on the ribosomes of the pathogen. Here, we describe how C. jejuni virulence is attenuated by altering the methylation of its ribosomes to disrupt the composition of its proteome, and how this in turn provides a means of identifying factors that are essential for infection and pathogenesis. Specifically, inactivation of the C. jejuni Cj0588/TlyA methyltransferase prevents methylation of nucleotide C1920 in the 23S rRNA of its ribosomes and reduces the pathogen’s ability to form biofilms, to attach, invade and survive in host cells, and to provoke the innate immune response. Mass spectrometric analyses of C. jejuni TlyA-minus strains revealed an array of subtle changes in the proteome composition. These included reduced amounts of the cytolethal distending toxin (CdtC) and the MlaEFD proteins connected with outer membrane vesicle (OMV) production. Inactivation of the cdtC and mlaEFD genes confirmed the importance of their encoded proteins in establishing infection. Collectively, the data identify a subset of genes required for the onset of human campylobacteriosis, and serve as a proof of principle for use of this approach in detecting proteins involved in bacterial pathogenesis.

Microbiological Assessment of Wheat and Buckwheat Sprouting Process

Introduction. Sprouted grain can cause food poisoning, since inappropriate conditions can promote the growth of pathogenic microorganisms on the grain surface. As a result, products of long-term storage use thermally-treated sprouted grain, the parameters of which depend on the initial bacteria content. There are different ways to reduce bacterial contamination of sprouted grain, each of which has its own advantages and disadvantages. Natural substances with antimicrobial properties, such as medicinal herbs, can serve as decontaminators. However, no scientific research has been performed so far to determine the exact temperature of grain sprouting to minimize its microbiological contamination. The research objective was to investigate the effect of antimicrobial agents and sprouting conditions on the microflora of wheat and buckwheat grain. Study objects and methods. The study featured wheat grain and green buckwheat grain. A set of experiments was performed to define the effect of antimicrobial agents and sprouting conditions on the quantity of mesophilic aerobic and facultative anaerobic microorganisms (QMAFAnM), molds, and yeasts. During sprouting at 10–30°C for 90 h, the grain was irrigated with distilled water, potassium permanganate solution (KMnO4), calendula infusion, and celandine i nfusion. QMAFAnM and the count of molds and yeasts were determined by standard methods; the qualitative analysis of the microflora was based on their morphological and cultural characteristics. Results and discussion. Microflora development during sprouting of wheat and buckwheat grains was controlled by selecting appropriate conditions and grain treatment methods. The herbal infusions for sprouting reduced the total microbial insemination of grain during sprouting by 52–68%; the calendula infusion reduced the contamination with molds by 47–51%, yeasts – by 100%. Conclusion. The research revealed the total microbial count and the count of mold and yeast colonies in dry sprouted grain. The optimal temperature of sprouting wheat and buckwheat was 20 ± 2°C in the infusion of medicinal herbs: it minimized the microflora of sprouted grain and reduced the sprouting time to 46 h. Calendula infusion could be recommended for commercial use in order reduce the microbiological contamination of sprouted grain. The initial microbial population of the product was found to affect the mode of heat treatment in long-term storage products.

Isolation of Shewanella putrefaciens GRD 03 from Fish and Explication of Biofilm Adherence Potency on Different Substrates

Foodborne pathogens are the main threat and cause of food poisoning. The majority of food infections have been related to the biofilm formation of foodborne pathogens in the food industry. Shewanella putrefaciens (KX355803, GRD 03), a Gram-negative pathogen isolated from mackerel fish, was identified and recognized as a food spoilage bacterium and a strong biofilm producer. The adhesion or attachment ability of Shewanella putrefaciens was determined on steel, plastic, glass, PVC and wood. NB (Nutrient broth), LB (Luria-Bertani broth), TSB (Tryptic soy broth) and BHI (Brain heart infusion broth) were enriched with glucose and shows optimum for bacterial adhesion. In the microtiter plate method (MTP), the strong attachment was observed at 48 and 72 hours of incubation and significant differences were obtained at p < 0.05. As the incubation period increases, the OD value (Optical density) of samples also increase. Biofilm formation is the major cause cross-contamination, and shows resistance to certain disinfectants, which leads to environmental stress tolerance. This study suggested with optimum biofilm production of isolate from fish by using glucose enriched media on different substrates, also comparing different growth media provide a detailed idea about biofilm-forming ability at different incubation time intervals.

Performance of distinct microbial based solutions in a Campylobacter infection challenge model in poultry

Background Antibiotic growth promoters (AGPs) are commonly used within poultry production to improve feed conversion, bird growth, and reduce morbidity and mortality from clinical and subclinical diseases. Due to the association between AGP usage and rising antimicrobial resistance, the industry has explored new strategies including the use of probiotics and other microbial-based interventions to promote the development of a healthy microbiome in birds and mitigate against infections associated with food safety and food security. While previous studies have largely focused on the ability of probiotics to protect against Clostridium perfringens and Salmonella enterica, much less is known concerning their impact on Campylobacter jejuni, a near commensal of the chicken gut microbiome that nevertheless is a major cause of food poisoning in humans. Results Here we compare the efficacy of four microbial interventions (two single strain probiotics, the bacterium—Pediococcus acidilactici, and the yeast—Saccharomyces cerevisiae boulardii; and two complex, competitive exclusion, consortia—Aviguard and CEL) to bacitracin, a commonly used AGP, to modulate chicken gut microbiota and subsequently impact C. jejuni infection in poultry. Cecal samples were harvested at 30- and 39-days post hatch to assess Campylobacter burden and examine their impact on the gut microbiota. While the different treatments did not significantly decrease C. jejuni burden relative to the untreated controls, both complex consortia resulted in significant decreases relative to treatment with bacitracin. Analysis of 16S rDNA profiles revealed a distinct microbial signature associated with each microbial intervention. For example, treatment with Aviguard and CEL increased the relative abundance of Bacteroidaceae and Rikenellaceae respectively. Furthermore, Aviguard promoted a less complex microbial community compared to other treatments. Conclusions Depending upon the individual needs of the producer, our results illustrate the potential of each microbial interventions to serve flock-specific requirements.

Prevalence of Bacillus cereus in Ready-to-Eat Foods in Northern Perak, Malaysia

Food poisoning cases in Malaysia showed an increasing trend every year where 496 episodes were reported in 2018 as compared to 401 episodes in the same week of the year 2017. Bacillus cereus is one of the foodborne pathogens related to food poisoning cases in Malaysia. The main cause for the outbreak of B. cereus is the unregulated temperature during holding time. This study was conducted to detect the presence of aerobic bacteria and B. cereus present in ready-to-eat food in Northern Perak. A total of 83 food samples were collected and analyzed for the microbial count. The result shows that aerobic bacteria and B. cereus were detected in 28% of the samples. B. cereus count in food samples tested ranged from 100 cfu/g to 42000 cfu/g, whereas the aerobic bacteria recorded a range of 500 cfu/g to 2100000 cfu/g. The highest percentage of B. cereus was found in rice-based food, followed by meat, poultry, and gravy dishes. Positive colonies of B. cereus were further tested for anti-microbial resistance profile. Most B. cereus isolates showed resistance to tetracycline and clindamycin.

Psychrotrophic properties, toxigenic characteristics, and PFGE profiles of Bacillus cereus isolated from different foods and spices

ABSTRACT: Bacillus cereus is an aerobic and facultatively anaerobic, spore-forming bacterium, and it is found naturally in soil and poses a risk factor for the contamination of food and foodstuffs including cereals, vegetables, spices, ready-to-eat (RTE) foods, meats, milk, and dairy products. This study determined the prevalence of B. cereus in raw poultry meat, raw cow’s milk, cheese, spices, and RTE foods in Hatay province. The study also analysed the psychrotrophic properties, toxigenic characteristics, and pulsed-field gel electrophoresis (PFGE) profiles of the isolates. The levels of contamination with B. cereus determined for cheese, raw milk, RTE foods, spices, and raw poultry meat were 16.6%, 34.2%, 42.8%, 49%, and 55.5%, respectively. B. cereus was isolated from 84 (42%) of the 200 samples analysed and the 84 isolates were verified by PCR analysis targeting the haemolysin gene specific for B. cereus. Of the total isolates, 64 (76.1%) were psychrotrophic. The toxin gene profiling of B. cereus isolates was determined by amplifying the four genes nhe, hbl, cytK, and ces. The nhe and cytK genes were most frequently detected in the isolates, while the hbl and ces genes were not found. In addition, a high genetic relationship between the isolates was detected at a 92% similarity level by PFGE analysis. In conclusion, the occurrence of both psychrotrophic and toxigenic B. cereus strains in this study indicated a potential risk for food spoilage and food poisoning.

In vitro study of antimicrobial activity of some plant seeds against bacterial strains causing food poisoning diseases

In this research, some plant seeds powder was evaluated to find their potential effect to rule diseases of food poisoning. Antimicrobial effect of five plant seeds was examined contra Bacillus cereus, Staphylococcus aureus, Escherichia coli, Klebsiella. pneumonia and Candida albicans by using well diffusion method. Antimicrobial activity studies revealed high potential activity of plant seeds powder of Nigella sativa L., cucurbita pepo, Sesamum radiatum, Trigonella foenum-graecum, Linum usitatissimum with variable efficiency contra tested microbial strains with concentration of 100 mg/ml, except Sesamum radiatum scored no effect. The T. foenum and N. sativa seed powder showed the largest inhibition zone (24-20 mm) contra K. pneumonia, followed by S. aureus (20-18 mm) and C. albicans (15mm) respectively. The five plant seeds powder exhibited bacteriostatic and bactericidal effects with MIC’s 20 and MBC 40 mg/ml against K. pneumonia, and MIC’s 40 and MBC 60 mg/ml against S. aureus. The results of this study indicated that plants seeds powder have promising antimicrobial activities and their potential applications in food process. It could be utilized as a natural medicinal alternative instead of chemical substance.

Assessment of Knowledge, Attitudes and Practices on the Risks of Food Poisoning in Collective Restaurants in the Kloto 1 Municipality, Togo

Collective food toxiinfections (CFTI) are frequent accidents, formidable by their health and economic consequences. They usually result from the consumption of food contaminated with microorganisms or chemical toxins. The objective of our study is to evaluate the knowledge, attitudes and practices of food product handlers in Kloto 1 municipality in order to guarantee food safety. To do this, we carried out a survey using google form whose questions were based on knowledge of foodborne diseases, good hygiene practices, good manufacturing practices, cross-contamination, wearing jewelry during handling, waste management. As a result of the study, It shows that during the years 2018, 2019 and 2020, the frequencies of toxiinfections recorded at the PHC of Kpalimé are respectively 33.09%, 41.84%, and 25.06%. To acces risk factor about this hight case of food poissoning in this area, 197 food handlers (FH) were interviewed and of these, 54.4% were women, 51.3% are between 15 and 25 years old, 67.2% have secondary education, and 68.7% have not received training on food safety. In terms of the level of knowledge, attitudes and practices, 10.7% of FH have insufficient knowledge about food safety, 98.0% have negative attitudes and 100.0% have insufficient food safety practices. Also, it shown that there is a strong positive correlation between knowledge and attitudes (rs = 0,267; p= 0.000), between knowledge and practices (rs = 0.182; p = 0.011), between attitudes and practices (rs = 0.253; p = 0.000), and between knowledge about foodborne illness and practice (rs = 0.189; p = 0,008). In short, the majority of FH have insufficient knowledge, practice and negative attitudes to avoid cases of CFTI because of the lack of training in food safety.