Evidence of sheep and abattoir environment as important reservoirs of multidrug resistant Salmonella and extended-spectrum beta-lactamase Escherichia coli.

The increase in antimicrobial-resistant (AMR) foodborne pathogens, including E. coli and Salmonella in animals, humans, and the environment, is a growing public health concern. Among animals, cattle, pigs, and chicken are reservoirs of these pathogens worldwide. There is a knowledge gap on the prevalence and AMR of foodborne pathogens in small ruminants (i.e., sheep and goats). This study investigates the prevalence and antimicrobial resistance of extended-spectrum beta-lactamase (ESBL) E. coli and Salmonella from sheep and their abattoir environment in North Carolina. We conducted a year-round serial cross-sectional study and collected a total of 1128 samples from sheep (n = 780) and their abattoir environment (n = 348). Sheep samples consisted of feces, cecal contents, carcass swabs, and abattoir resting area feces. Environmental samples consisted of soil samples, lairage swab, animal feed, and drinking water for animals. We used CHROMAgar EEC with 4 µg/ml of Cefotaxime for isolating ESBL E. coli, and ESBL production was confirmed by double-disk diffusion test. Salmonella was isolated and confirmed using standard methods. All of the confirmed isolates were tested against a panel of 14 antimicrobials to elucidate susceptibility profiles. The prevalence of ESBL E. coli and Salmonella was significantly higher in environmental samples (47.7% and 65.5%) compared to the sheep samples (19.5% and 17.9%), respectively (P < 0.0001). We recovered 318 ESBL E. coli and 368 Salmonella isolates from sheep and environmental samples. More than 97% (310/318) of ESBL E. coli were multidrug-resistant (MDR; resistant to ≥3 classes of antimicrobials). Most Salmonella isolates (77.2%, 284/368) were pansusceptible, and 10.1% (37/368) were MDR. We identified a total of 24 different Salmonella serotypes by whole genome sequencing (WGS). The most common serotypes were Agona (19.8%), Typhimurium (16.2%), Cannstatt (13.2%), Reading (13.2%), and Anatum (9.6%). Prevalence and percent resistance of ESBL E. coli and Salmonella isolates varied significantly by season and sample type (P < 0.0001). The co-existence of ESBL E. coli in the same sample was associated with increased percent resistance of Salmonella to Ampicillin, Chloramphenicol, Sulfisoxazole, Streptomycin, and Tetracycline. We presumed that the abattoir environment might have played a great role in the persistence and dissemination of resistant bacteria to sheep as they arrive at the abattoir. In conclusion, our study reaffirms that sheep and their abattoir environment act as important reservoirs of AMR ESBL E. coli and MDR Salmonella in the U.S. Further studies are required to determine associated public health risks.

Containment of antimicrobial resistance due to use of antimicrobial agents in animals intended for food: WHO perspective.

The use of antimicrobial agents in humans and food-producing animals has important consequences for human and animal health, as it can lead to the development of resistant bacteria (pathogens and/or commensals with resistance genes). Moreover, resistant bacteria in animals can be transferred to people--usually through the consumption of food, but also through direct contact with food-producing animals or through environmental spread. Ultimately, this can result in human infections with bacteria that are resistant to antimicrobial agents and that can therefore be difficult or impossible to cure. Of special concern is resistance to antimicrobial agents classified by the World Health Organization (WHO) as critically important for human medicine, such as fluoroquinolones, third- and fourth-generation cephalosporins, and macrolides. WHO encourages the agricultural, food, veterinary and health sectors to work together to eliminate the burden of antimicrobial resistance arising from the use of antimicrobial agents in food-producing animals. Joint efforts should be made to reduce the inappropriate use of antimicrobial agents (e.g. the use of antimicrobials as growth promoters) and limit the spread of bacteria resistant to antimicrobial agents. WHO will continueto address this issue in conjunction with the Food and Agriculture Organization of the United Nations, the World Organisation for Animal Health, the animal health/production industry and other important stakeholders. It will also continue to enhance the capacity of its Member States (through training courses and sentinel studies), particularly developing countries, to conduct integrated surveillance of antimicrobial use and resistance, to carry out risk assessments to support the selection of risk management options and to implement strategies for the containment of antimicrobial resistance.

Genotypic Characterization of Antibiotic-Resistant Salmonella Enteritidis Isolates in Dakar; Senegal

Background: It is well established that Salmonella enterica is a major cause of food-borne disease worldwide. In Africa; according to the Who Global Salm-Surv country data bank from 2000 to 2002 Salmonella enterica serovar Enteritidis was the most common serotype involved in human salmonellosis. In Dakar this serotype of Salmonella has been reported as a frequent and an increasing cause of human infection. Methodology: The genetic determinants of the antimicrobial resistance of 25 selected multiresistant strains of Salmonella enterica serovar Enteritidis referred to the National Reference Center for Enterobacteria (NRCE) in Dakar were investigated using molecular techniques. Results: All strains carried blaTEM 1 genes. Five harboured three types of class 1 integrons with gene cassettes dfrA15; dfrA1- aadA1 and dfrA7. Multiresistance was due to a 23 Kb conjugative plasmid. DNA fingerprinting by macrorestriction of genomic DNA revealed a single related group suggesting that strains might be clonal. Conclusions: The spread of resistance genes through plasmid transfer plays an important role in the dissemination of antibiotic resistance in enteric pathogens such as Salmonella Enteritidis; the risk of transmissibility of antibiotic resistance between different bacterial strains highlights the urgent need to develop strategies to limit the spread of antimicrobial resistance among bacterial enteropathogens

Class 2 integron-associated antibiotic resistance in Shigella sonnei isolates in Dakar, Senegal.

Forty-three Shigella sonnei isolates from adult patients with diarrhoea in Dakar were analysed for the presence of integrons. Isolates were resistant to sulphamethoxazole, trimethoprim, tetracycline, streptomycin and spectinomycin. A high prevalence of class 2 integrons (93%) was found. These integrons showed three distinct structures: a class 2 integron, part of the Tn7 family and its derivatives, carrying four cassettes in the order dfrA1-sat-aadA1-orfX; a truncated class 2 integron, without orfX; and a third type ca. 4 kb in size. These class 2 integrons probably play a role in the spread of multiresistance in S. sonnei isolates. To our knowledge, this is the first description of class 2 integrons in S. sonnei isolated in sub-Saharan Africa.

Ordinary and opportunistic enteropathogens associated with diarrhea in Senegalese adults in relation to human immunodeficiency virus serostatus.

OBJECTIVES: A survey was conducted in Dakar, Senegal, to identify major types and prevalences of bacteria, parasites, fungi, and Rotaviruses associated with diarrhea in relation to human immunodeficiency virus (HIV) serostatus with the goal to provide guidance to physicians for case management. METHODS: Etiologic agents were identified in a case control study: cases were HIV-infected patients with diarrhea (HIV+ D+) and HIV seronegative patients with diarrhea (HIV D+); controls were HIV-infected patients without diarrhea (HIV+ D ) and seronegative controls without diarrhea (HID D ). Ordinary enteric pathogens were identified by conventional methods. Different Escherichia coli pathotypes were characterized by polymerase chain reaction (PCR), identification of HEp-2 cell adherence pattern, Sereny test, GM1-ELISA, and the suckling mouse assay. Opportunistic parasites, such as Cryptosporidium and Microsporidium, were identified by the Kinyoun method and trichromic stain of Weber, respectively. Rotaviruses were identified with a commercial latex agglutination kit. Antimicrobial susceptibility testing was carried out by the disk diffusion method. RESULTS: Among the 594 patients examined, 158 were HIV+ D+, 121 were HIV2 D+, 160 were HIV+ D , and 155 were HIV D . The main etiologies of diarrhea were different according to HIV serostatus of patients. In immunocompetent adults the main causes of diarrhea were Shigella sp (12.4%), Entamoeba histolytica(10.7%), Salmonella enterica (6.6%), and Giardia (4.9%). In the immunocompromised host the more frequent pathogens were enteroaggregative E. coli (19.6%), Microsporidium (9.4%), Cryptosporidium sp (8.2%), Rotavirus (8.2%), Shigella sp (7.6%), Candida albicans (7.6%), E. histolytica (5.1%), S. enterica (4.4%), and Isospora belli (4.4%). Also, Blastocystis hominis has to be considered as an opportunistic parasite, because it was identified only in HIV-infected patients, with higher prevalence in adults with diarrhea (2.5% in HIV+ D+ patients; 0.6% in HIV+ D patients). High level of asymptomatic carriage of Ascaris lumbricoides and Trichuris trichiura and some cases of multiple infections were observed. Fungi, Cryptosporidium sp and Microsporidium sp, were often identified in patients with low CD4 counts (range, 79 250 cells/mL). Independently from HIV-serostatus, CD4 count was lower in diarrheic persons, suggesting that diarrhea is a debilitating illness and that effective management of diarrhea can prevent immunosuppression. Isolated enteropathogenic strains displayed high resistance to most antibiotics used in Senegal for treating diarrhea (ampicillin, tetracycline, cotrimoxazole); they were susceptible to amikacin, gentamicin, and norfloxacin. CONCLUSION: These epidemiologic data suggest that guidelines for the management of diarrhea during HIV infection in Dakar should be updated.

[Infective etiology of diarrhea in adults with HIV infection in Dakar: a case-control study on 594 patients]. / Etiologies infectieuses des diarrhées de l'adulte au cours de l'infection à VIH à Dakar: étude cas/témoins sur 594 malades.

A survey was conducted in Dakar, Senegal from May 1997 to May 1999, to identify major types and prevalences of bacteria, parasites, fungi and Rotaviruses associated with diarrhea in relation to human immunodeficiency virus serostatus with the aim to provide guidance to physicians for case management. Etiologic agents were identified in a case-control study: cases were HIV-infected patients with diarrhea (HIV+ D+) and HIV seronegative patients with diarrhea (HIV- D+); controls were HIV-infected patients without diarrhea (HIV+ D-) and seronegative controls without diarrhea (HIV- D-). Strict enteric pathogens were identified by conventional methods. Different E. coli pathotypes were characterized by PCR. Opportunistic parasites such as Cryptosporidium and Microsporidium were identified by the Kinyoun method and trichromic stain of Weber respectively. Rotaviruses were identified with a commercial latex agglutination kit. Antimicrobial susceptibility testing was carried out by the disk diffusion method. A total of 594 patients were examined (158 HIV+D+, 121 HIV- D+, 160 HIV+ D- and 155 HIV- D-). In immunocompetent adults the main causes of diarrhea were: Shigella sp. (12.4%), Entamoeba histolytica (10.7%), Salmonella enterica (6,6%) and Giardia lamblia (4.9%). In the immunocompromised host the more frequent pathogens were: enteroaggregative E.coli (19,6%), Microsporidium(9.4%), Cryptosporidium sp.(8.2%), Rotavirus (8.2%), Shigella sp. (7.6%), Candida albicans (7.6%), Entamoeba histolytica (5,1%), Salmonella enterica (4.4%), Isospora belli (4.4%) and Blastocystis hominis (2,5%). Isolated enteropathogenic strains displayed high resistance to most antibiotics used in Senegal for treating diarrhea (tetracycline, cotrimoxazole); they were susceptible to quinolons and cephalosporins III .

[Microbiological quality of street-vendor ice cream in Dakar]. / Qualité microbiologique des glaces vendues sur la voie publique à Dakar.

During a multicenter study initiated by the International Network of Pasteur Institutes and Associated Institutes, microbiological quality of street-vended ice creams in Dakar was evaluated. 313 samples of ice creams from 170 street-vendors were collected and tested for common foodborne pathogens and indicator organisms. Results showed that microbiological quality of 45% of tested samples was unsatisfactory because of large populations of aerobic mesophilic organisms (36.7%), thermotolerant coliforms bacteria (21.4%) and sometimes E. coli. (10.6%). Strict pathogens as Salmonella, Shigella and Vibrio cholerae were not found. An investigation conducted among vendors showed a lack of education and training; these vendors need information about food preparation and storage practices that reduce microbiological contamination of foods.

[Characterization of entomopathogenic Bacillus samples isolated in Senegal and study of their toxicity for malaria vectors]. / Caractérisation de souches de Bacillus entomopathogènes isolées au Sénégal et étude de leur toxicité pour les vecteurs du paludisme.

A screening program developed in Senegal to isolate new strains of entomopathogenic Bacillus has led to the isolation of 194 strains of Bacillus thuringiensis and 9 strains of Bacillus sphaericus from various sites and insect samples. The characterization of these strains regarding their H serotype, their crystal composition and their toxicity against mosquitoes (Culex pipiens, Aedes aegypti and Anopheles stephensi) has led to the isolation of 27 mosquitocidal strains. As malaria is an important public health problem in Senegal, these strains were more completely characterized looking for their toxicity against the two major malaria vectors in Senegal: Anopheles gambiae and Anopheles arabiensis.

Isolation of a new variant of Vibrio cholerae O1: V. cholerae O1 ribotype B27 toxinogenotype TB31 during the last cholera epidemic in Senegal.

A total of 205 Vibrio cholerae O1 isolates from recent cholera epidemic in Senegal were analyzed by conventional methods, polymerase chain reaction (PCR) for genes encoding cholera toxin (ctx A), zonula occludens toxin (zot) and accessory cholera enterotoxin (ace), ribotyping and toxinogenotyping. Ribotyping after Bg1 I digestion of total DNA revealed that ribotype B5a, the predominant ribotype of the seventh pandemic in Africa and Asia, was not isolated. A new ribotype designated B27 in our database is predominant and was associated with a new toxinogenotype designated TB31.