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The purpose of this paper is to evaluate the frequency and antimicrobial susceptibility pattern of pathogens present in ready-to-eat salads available at a local market.

A 100 salad samples were collected aseptically. Each sample (25 g) was homogenized in 225 ml of sterile peptone water and was serially diluted up to 1×106. Dilutions were inoculated on nutrient agar by surface spread plate technique. Aerobic colony count (ACC) was estimated by counting the colonies. Bacterial isolates were cultured on blood and MacConkey agar and identified on the basis of their morphology, culture characteristics and confirmed by API 20E and 20NE. Antimicrobial susceptibility was determined as per CLSI 2014.

ACC range was 1.1×103 cfu/g to 5.8×109 cfu/g. Among these the highest ACC was found in channa chat (4.9×104 to 5.8×109 cfu/g). A total of 127 microorganisms were identified; 73 were gram negative rods (GNRs) and 24 were gram positive cocci (GPC). Among GNRs; Klebsiella spp. (n=18) was the predominant whereas among GPC, Staphylococcus aureus (n=6) were the chief pathogen. Klebsiella spp. showed 100 percent resistance to ampicillin, 89-78 percent to amoxicillin/clavulanic acid and 33 percent to imipenem, however, Enterobacter spp. were resistant to ampicillin (100 percent) amoxicillin/clavulanic acid (77 percent) and imipenem (23 percent). Staphylococcus aureus showed resistance to co-amoxiclav (83 percent) and penicillin (75 percent).

This study revealed that effective control measures must been implemented and respected by throughout the entire food chain and better surveillance studies should be performed at national level to reduce the spread of bacteria by fresh salads.

This paper explore the high prevalence of multidrug-resistant pathogens in different salads and most of the salads were found to be unhygienic for consumption.

The objective of this study was to determine Salmonella prevalence, antimicrobial-resistant phenotypes, and their genetic relatedness in frozen organic chicken collected at retail level in Turkey.

Retail packs (n = 348) of cut-up chicken parts (breast, leg quarter and drumstick) and whole chicken carcasses were purchased from a central hypermarket in Diyarbakir (Southeast Anatolia Region in Turkey) and from a large online retailer in Turkey. The retail packs were paired by part type, brand, production date, and sell-by date. The chicken samples were analyzed for the presence of Salmonella spp., and then isolates were screened for antibiotic susceptibility, class I integron, and genetic similarity.

Salmonella prevalence in retail frozen organic chicken samples was 6.3 percent; however, the prevalence by parts, leg quarter, drumstick, breast, and whole chicken was 2.1 percent, 10.4 percent, 10.4 percent, and 0 percent, respectively. Salmonella prevalence was significantly higher in samples obtained from the hypermarket (9.2 percent) compared to online retailer (3.8 percent). All the isolates were serotype Infantis, genetically similar (highly clonal), and 68.2 percent harbored class I integron. All isolates were resistant to ciprofloxacin (drug of choice to treat salmonellosis in human), and 86.3 percent of the isolates were multidrug-resistant.

Salmonella prevalence in organic chicken meat, regardless of the retail market source in Turkey, may pose a health risk to consumers especially with the high prevalence of multi-drug resistant phenotypes. Findings inform researchers and the public about the safety of organically produced chicken and the potential health risk to consumers.

The purpose of this paper is to evaluate the prevalence and antibiograms of bacteria isolated from various fresh fruit juices at a local market in Faisalabad.

Fresh fruit juice samples (n=125) were randomly collected using aseptic technique. Each sample (10 mL) was serially diluted with 90 mL of sterile peptone water, from 1×10⁻¹ to 1×10⁻⁵. Each dilution was then used to inoculate nutrient agar by surface spread plating. Aerobic colony counts (ACCs) were determined by colony counting. The isolates were sub-cultured on blood and MacConkey agar. Preliminary identification was achieved on the basis of colony morphology and culture characteristic, and confirmed by API® 20E, 20NE, and API® Staph testing. Antimicrobial susceptibility testing was carried out using the Kirby-Bauer disk diffusion assay, per CLSI 2015 guidelines.

The mean ACC ranged from 2.0×10⁶ CFU/mL to 4.93×10⁶ CFU/mL, with the highest ACC determined for orange juice. Overall, 153 polymicrobial were identified in 125 samples; 103 of these were Gram-negative rods (GNR) and 28 were Gram-positive cocci (GPC). Escherichia coli (n=38), Klebsiella pneumoniae (n=32) and Pseudomonas aeruginosa (n=24) were the predominant GNR; Staphylococcus aureus (n=28) was the predominant GPC. Antibiogram analysis revealed that all GNR were resistant to ampicillin. However, most E. coli isolates were resistant to ceftazidime (72.4 percent of isolates), and ceftriaxone and cefepime (68.9 percent), while most K. pneumoniae isolates were resistant to cefepime (72 percent) and ceftriaxone (64 percent). All S. aureus isolates were resistant to penicillin, while most (64 percent) were resistant to piperacillin; the most effective drugs against bacteria were vancomycin and imipenem.

The findings suggest that the local government regulatory food and public health authorities should take immediate emergency measures. Appropriate surveillance studies and periodic monitoring of food items should be regularly performed to safeguard public health.

The current study revealed the prevalence of multidrug-resistant bacteria in freshly prepared fruit juices sold by local street vendors.

This chapter provides a brief overview of our understanding of major public health challenges and environmental concerns in Karakalpakstan today, and highlights questions that still remain unanswered. As seen in the case of Muynak, the fishing town on the southern edge of the former Aral Sea, ecological disasters do not happen alone – they spur socioeconomic disasters that only heighten the health disasters. The loss of the sea, the loss of local livelihoods, and mass out-migration of the population, along with economic depression following the collapse of the Soviet Union, have adversely affected the community living in Muynak. They face major public health challenges, such as tuberculosis, multidrug resistant tuberculosis, and anemia as a result of their impoverishment. The desiccation of the Aral Sea is but one of the many disasters linked to intensive cotton cultivation in Uzbekistan. Pesticide contamination and the salinization of drinking water in Karakalpakstan are yet other environmental disasters that further threaten the health of the population and of future generations. Currently, there is an urgent need for greater international involvement and collaboration with Uzbeks to reverse the poor public health trends and to study the extent of environmental contamination in communities across Karakalpakstan, in order to reduce the health threats presented by these.

The purpose of this paper is to assess the occurrence of the extended-spectrum β-lactamases (ESBLs) and AmpC β-lactamase genes in 144 Escherichia coli isolates recovered from 160 vegetable salad samples.

Among the 144 E. coli isolates recovered from 160 vegetable salads, 17 (12 percent) ceftazidime-resistant isolates were screened for ESBL production with the double disk-diffusion test. The ESBL-producing isolates were characterized for antimicrobial resistance, the presence of virulence genes and plasmid-mediated quinolone resistance (PMQR) determinants. The isolates were also subjected to phylogenetic group typing. The existence of plasmid AmpC genes and mutations in the regulatory region of the chromosomal AmpC gene was assessed using polymerase chain reaction (PCR) and sequencing. All β-lactamase isolates were further characterized by pulsed-field gel electrophoresis to determine the genetic relatedness.

Overall, 17 (12 percent) of the 144 E. coli isolates studied were ceftazidime resistant. Among the 17 isolates, 13 (77 percent) were multidrug resistant and four (23.5 percent) were ESBL producers. The bla CTX-M14 was the only gene detected. Of the 12 AmpC-producing isolates, three (18 percent) harbored plasmid-encoded AmpC and sequencing analysis of the chromosomal AmpC genes revealed mutations in the promoter/attenuator region. PMQR determinants were detected in 9 (52 percent) isolates. A was the most prevalent phylogenetic group (56 percent), followed by groups B1 (31 percent), D (6 percent), and B2 (6 percent). PCR showed that six (50 percent) ESBL/AmpC-producing E. coli isolates carried one and/or two virulence genes. Pulsed-field gel electrophoresis showed no epidemiological relationship between these isolates.

This study places vegetable salads within the spectrum of ecological niches that may be vehicles for antibiotic-resistant bacteria/genes with clinical interest and these findings are worthy of attention as their spread to humans by ingestion cannot be dismissed.

Broad-spectrum cephalosporin resistance is rapidly increasing in Escherichia coli, representing a food safety problem. The purpose of this paper is to characterize eight extended-spectrum-ß-lactamase (ESBL) and acquired AmpC ß-lactamase-producing E. coli isolates and virotypes associated, obtained from chicken and pork food samples in Puebla, Mexico.

Samples (36 from chicken and 10 from pork) were cultured on Levine agar plates supplemented with cefotaxime (2 mg/L) for isolation of cefotaxime-resistant (CTXR) E. coli. CTXR-E. coli isolates were detected in 33 of 46 samples (72 percent), and one isolate/sample was characterized (28 from chicken and 5 from pork), for ESBL production, phylogenetic group, sequence typing, resistance and virulence genes by PCR and sequencing.

Results showed 16 ESBL-E. coli (35 percent) (12/16 belonging to phylogroup B1) and 8 CMY-2-E. coli (17 percent). ESBL detected were as follows (number of isolates): CTX-M-2 (8); CTX-M-1 (2); CTX-M-15 (1); SHV-2a (4) and TEM-52c (1). In total, 20 different sequence types (STs) were identified among the ESBL- or CMY-2-producing E. coli strains, which included four new ones. The CTX-M-15 β-lactamase was detected in one E. coli ST617-ST10 Cplx-B1 strain that also carried ibeA gene. One CMY-2-positive strain of lineage ST224-B2 was detected and it carried the qnrA1 gene.

In this study, a ST131-based virotyping scheme for strains from food of animal origin was established since this kind of strains constitutes an important vehicle of virulent ESBL- and CMY-2-producing E. coli isolates, which could be transmitted to humans by direct contact or through the food chain.

Antimicrobial resistance is a public health threat even in countries exercising aggressive antimicrobial stewardship. A market failure is also causing lackluster innovation in antimicrobial medicines development. At the heart of the issue are antimicrobial stewardship guidelines that, rightfully, reserve innovative antimicrobials for emergency situations that arise due to multidrug-resistant organisms. This suppresses revenues and research and development (R&D) investment incentives of manufacturers. The public policy makers and researchers have taken aim at the problem. The researchers have published strategies to encourage the production of innovative antimicrobials, while policy makers have taken legislative steps to address the issue. Most notably, the USA enacted the Generating Antibiotic Incentives Now (GAIN) act in 2012 and the EU created a commission to formally study possible policy solutions. The paper aims to discuss these issues.

In this paper, the authors describe incentives that drive pharmaceutical R&D and review the impact of a number of R&D stimulus policies in other pharmaceutical markets. The authors also discuss which policy levers are useful to boost R&D of new antimicrobials.

The authors find that a policy focused on extending intellectual property rights, as implemented in the GAIN act, are unlikely to be impactful. Instead, the authors see a need for the revision of the procurement policy to move away from paying per prescription and toward licenses and advanced market commitment models. Further, the authors note that the importance of steadfast public investment in basic biomedical research as it has been repeatedly shown to boost innovation.

The authors hope that the work can support the refinement of the GAIN act and the EU efforts.

This study aims to study the distribution characteristics of antibiotic resistance in direct-eating food and analysis of Citrobacter freundii genome and pathogenicity. Residual antibiotics and antibiotic resistance genes (ARGs) in the environment severely threaten human health and the ecological environment. The diseases caused by foodborne pathogenic bacteria are increasing daily, and the enhancement of antibiotic resistance of pathogenic bacteria poses many difficulties in the treatment of disease.

In this study, six fresh fruits and vegetable samples were selected for isolation and identification of culturable bacteria and analysis of antibiotic resistance. The whole genome of Citrobacter freundii isolated from cucumber was sequenced and analyzed by Oxford Nanopore sequencing.

The results show that 270 strains of bacteria were identified in 6 samples. From 12 samples of direct food, 2 kinds of probiotics and 10 kinds of opportunistic pathogens were screened. The proportion of Citrobacter freundii screened from cucumber was significantly higher than that from other samples, and it showed resistance to a variety of antibiotics. Whole genome sequencing showed that Citrobacter freundii was composed of a circular chromosome containing signal peptides, transmembrane proteins and transporters that could induce antibiotic efflux, indicating that Citrobacter freundii had strong adaptability to the environment. The detection of genes encoding carbohydrate active enzymes is more beneficial to the growth and reproduction of Citrobacter freundii in crops. A total of 29 kinds of ARGs were detected in Citrobacter freundii, mainly conferring resistance to fluoroquinolones, aminoglycosides, carbapenem, cephalosporins and macrolides. The main mechanisms are the change in antibiotic targets and efflux pumps, the change in cell permeability and the inactivation of antibiotics and the detection of virulence factors and ARGs, further indicating the serious risk to human health.

The detection of genomic islands and prophages increases the risk of horizontal transfer of virulence factors and ARGs, which spreads the drug resistance of bacteria and pathogenic bacteria more widely.

The Chulalongkorn-Hasanuddin Rifampicin-Resistant Tuberculosis Screening Tool (CUHAS-ROBUST) is an artificial intelligence–based (AI–based) application for rifampicin-resistant tuberculosis (RR-TB) screening. This study aims to elaborate on the drug-resistant TB (DR-TB) problem and the impact of CUHAS-ROBUST implementation on RR-TB screening.

A qualitative approach with content analysis was performed from September 2020 to October 2020. Medical staff from the primary care center were invited online for application trials and in-depth video call interviews. Transcripts were derived as a data source. An inductive thematic data saturation technique was conducted. Descriptive data of participants, user experience and the impact on the health service were summarized

A total of 33 participants were selected from eight major islands in Indonesia. The findings show that DR-TB is a new threat, and its diagnosis faces obstacles particularly prolonged waiting time and inevitable delayed treatment. Despite overcoming the RR-TB screening problems with fast prediction, the dubious screening performance, and the reliability of data collection for input parameters were the main concerns of CUHAS-ROBUST. Nevertheless, this application increases the confidence in decision-making, promotes medical procedure compliance, active surveillance and enhancing a low-cost screening approach.

The CUHAS-ROBUST achieved its purpose as a tool for clinical decision-making in RR-TB screening. Moreover, this study demonstrates AI roles in enhancing health-care quality and boost public health efforts against tuberculosis.