The aquatic ecosystem, a good environment for the horizontal transfer of antimicrobial resistance and virulence-associated factors among extended spectrum beta-lactamases producing E. coli

One of the main public health problems nowadays is the increase of antimicrobial resistance, both in the hospital environment and outside it (animal environment, food and aquatic ecosystems, among others). It is necessary to investigate the virulence-associated factors and the ability of horizontal gene transfer among bacteria for a better understanding of the pathogenicity and the mechanisms of dissemination of resistant bacteria. Therefore, the objective of this work was to detect several virulence factors genes (fimA, papC, papG III, cnf1, hlyA and aer) and to determine the conjugative capacity in a wide collection of extended-spectrum β-lactamases-producing E. coli isolated from different sources (human, food, farms, rivers, and wastewater treatment plants). Regarding virulence genes, fimA, papC, and aer were distributed throughout all the studied environments, papG III was mostly related to clinical strains and wastewater is a route of dissemination for cnf1 and hlyA. Strains isolated from aquatic environments showed an average conjugation frequencies of 1.15 × 10−1 ± 5 × 10−1 , being significantly higher than those observed in strains isolated from farms and food (p < 0.05), with frequencies of 1.53 × 10−4 ± 2.85 × 10−4 and 9.61 × 10−4 ± 1.96 × 10−3 , respectively. The reported data suggest the importance that the aquatic environment (especially WWTPs) acquires for the exchange of genes and the dispersion of resistance. Therefore, specific surveillance programs of AMR indicators in wastewaters from animal or human origin are needed, in order to apply sanitation measures to reduce the burden of resistant bacteria arriving to risky environments as WWTPs

The aquatic ecosystem, a good environment for the horizontal transfer of antimicrobial resistance and virulence-associated factors among extended spectrum ß-lactamases producing E. coli

One of the main public health problems nowadays is the increase of antimicrobial resistance, both in the hospital environment and outside it (animal environment, food and aquatic ecosystems, among others). It is necessary to investigate the virulence-associated factors and the ability of horizontal gene transfer among bacteria for a better understanding of the pathogenicity and the mechanisms of dissemination of resistant bacteria. Therefore, the objective of this work was to detect several virulence factors genes (fimA, papC, papG III, cnf1, hlyA and aer) and to determine the conjugative capacity in a wide collection of extended-spectrum β-lactamases-producing E. coli isolated from different sources (human, food, farms, rivers, and wastewater treatment plants). Regarding virulence genes, fimA, papC, and aer were distributed throughout all the studied environments, papG III was mostly related to clinical strains and wastewater is a route of dissemination for cnf1 and hlyA. Strains isolated from aquatic environments showed an average conjugation frequencies of 1.15 × 10−1 ± 5 × 10−1 , being significantly higher than those observed in strains isolated from farms and food (p < 0.05), with frequencies of 1.53 × 10−4 ± 2.85 × 10−4 and 9.61 × 10−4 ± 1.96 × 10−3 , respectively. The reported data suggest the importance that the aquatic environment (especially WWTPs) acquires for the exchange of genes and the dispersion of resistance. Therefore, specific surveillance programs of AMR indicators in wastewaters from animal or human origin are needed, in order to apply sanitation measures to reduce the burden of resistant bacteria arriving to risky environments as WWTPs

Listeriosis y poblaciones de riesgo

Listeria monocytogenes es una bacteria patógena para el hombre que se transmite principalmente a través de los alimentos. Se trata de un patógeno emergente del cual se notifican cada año más casos a nivel mundial. El objetivo de este trabajo es analizar la incidencia de listeriosis en EEUU y Europa durante los últimos 15 años con el fin de conocer el impacto de la enfermedad en las poblaciones de riesgo, y determinar los principales grupos de alimentos implicados en la transmisión de este patógeno. La revisión bibliográfica se realizó a través de la base de datos Pubmed, introduciendo los descriptores y operadores boleanos siguientes: “Infections, Listeria” AND “Risk group”, y “Listeriosis” AND “Food”. Los criterios utilizados para seleccionar los artículos fueron: últimos 15 años, datos procedentes de EEUU y Europa, acceso al texto completo en inglés o castellano. Por otro lado, se consultaron páginas webs de Organismos oficiales para recopilar datos epidemiológicos y de alertas alimentarias. Los resultados del análisis mostraron una mayor incidencia de listeriosis en España frente a la media de países europeos, con tasas máximas en 2018 de 1,07 vs 0,47 casos/100.000 habitantes, respectivamente. La población de riesgo con mayor incidencia son las personas mayores de 65 años (69,1% de los casos de listeriosis en 2018). Sin embargo, las personas inmunodeprimidas son las que mayor ratio de letalidad tienen (ratio máximo de 43% en cáncer de páncreas) En el caso de mujeres embarazadas existe mayor probabilidad de supervivencia del feto si la infección ocurre en el 3er trimestre. La mayoría de infecciones ocurren en el hogar (75-98%) y requieren de hospitalizaciones (84,6-99,1%). Los grupos de alimentos con mayor prevalencia de L. monocytogenes son los pescados y carnes y sus derivados Ready-to-eat (RTE) con valores entre 12,1-19,5%. En conclusión, se confirma la tendencia al alza de los casos de listeriosis y se prevé un crecimiento mayor debido al aumento de esperanza de vida (el 65% de los casos ocurren en mayores de 65 años) y al mayor consumo de alimentos RTE. La elevada presencia del patógeno en estos alimentos y su capacidad de multiplicarse en refrigeración, hace necesario reforzar la formación de los manipuladores de alimentos y aportar recomendaciones dietéticas a la población en riesgo para reducir los casos de listeriosis.Listeria monocytogenes is a human pathogenic bacteria, acquiered principally by the ingestion of contaminated food. It is a rising pathogen and every year are more notified cases around the world. The objective of this paper is to analyse the incidence of listeriosis in USA and Europe during the fifteen last years, with the aim of known the impact in the risk groups and to determine the main food group involved in the transmission of this pathogen. The bibliographic revision was done using Pubmed database with the followings descriptors and Mesh terms: “Infections, Listeria” AND “Risk group”, and “Listeriosis” AND “Food”. The inclusive criteria used were: last fifteen years, data from USA and Europe, full text in English and Spanish language. Moreover, it was consulted official webs in order to gather epidemiological and food alerts data. The outcomes of the analysis evidenced that the incidence of listeriosis in Spain is bigger than de mean of European countries, as it is reflected in the reported tases in 2018, 1.07 vs 0.47 cases/100,000, respectively. Elderly risk group has the most incidence with 69.1% of the total cases in 2018. However, immunodeficient people have the most lethality ratio, been pancreas cancer the worst, with a value of 43%. Infection´s pregnant women during the 3er trimester, has 95% of viability for the foetus. Majority of the infections occur at home (75-98%) and require hospitalisation 84.6-99.1% of the cases. The food groups with the higher prevalence of pathogen are fish and meat and their derived ready-to-eat products (RTE), with values between 12.1-19.5%. In conclusion, listeriosis incidence is rising and it seems to being more in the coming years due to the increasing life expectancy (65% cases happen in people older than 65 years) and the increasing consumption of food RTE. Due to the high presence of L. monocytogenes in this products and its ability to grow in refrigeration conditions, make necessary to improve the training of food workers and to provide dietetic´s advices to the risk group, in order to decrease the listeriosis cases

Multidrug-Resistant Bacteria Isolated from Different Aquatic Environments in the North of Spain and South of France

Due to the global progress of antimicrobial resistance, the World Health Organization (WHO) published the list of the antibiotic-resistant “priority pathogens” in order to promote research and development of new antibiotics to the families of bacteria that cause severe and often deadly infections. In the framework of the One Health approach, the surveillance of these pathogens in different environments should be implemented in order to analyze their spread and the potential risk of transmission of antibiotic resistances by food and water. Therefore, the objective of this work was to determine the presence of high and critical priority pathogens included in the aforementioned list in different aquatic environments in the POCTEFA area (North Spain–South France). In addition to these pathogens, detection of colistin-resistant Enterobacteriaceae was included due its relevance as being the antibiotic of choice to treat infections caused by multidrug resistant bacteria (MDR). From the total of 80 analyzed samples, 100% of the wastewater treatment plants (WWTPs) and collectors (from hospitals and slaughterhouses) and 96.4% of the rivers, carried antibiotic resistant bacteria (ARB) against the tested antibiotics. Fifty-five (17.7%) of the isolates were identified as target microorganisms (high and critical priority pathogens of WHO list) and 58.2% (n = 32) of them came from WWTPs and collectors. Phenotypic and genotypic characterization showed that 96.4% were MDR and resistance to penicillins/cephalosporins was the most widespread. The presence of bla genes, KPC-type carbapenemases, mcr-1 and vanB genes has been confirmed. In summary, the presence of clinically relevant MDR bacteria in the studied aquatic environments demonstrates the need to improve surveillance and treatments of wastewaters from slaughterhouses, hospitals and WWTPs, in order to minimize the dispersion of resistance through the effluents of these areas

Prevalence of integrons and insertion sequences in ESBL-producing E-coli isolated from different sources in Navarra, Spain

Mobile genetic elements play an important role in the dissemination of antibiotic resistant bacteria among human and environmental sources. Therefore, the aim of this study was to determine the occurrence and patterns of integrons and insertion sequences of extended-spectrum β-lactamase (ESBL)-producing Escherichia coli isolated from different sources in Navarra, northern Spain. A total of 150 isolates coming from food products, farms and feeds, aquatic environments, and humans (healthy people and hospital inpatients), were analyzed. PCRs were applied for the study of class 1, 2, and 3 integrons (intI1, intI2, and intI3), as well as for the determination of insertion sequences (IS26, ISEcp1, ISCR1, and IS903). Results show the wide presence and dissemination of intI1 (92%), while intI3 was not detected. It is remarkable, the prevalence of intI2 among food isolates, as well as the co-existence of class 1 and class 2 (8% of isolates). The majority of isolates have two or three IS elements, with the most common being IS26 (99.4%). The genetic pattern IS26–ISEcp1 (related with the pathogen clone ST131) was present in the 22% of isolates (including human isolates). In addition, the combination ISEcp1–IS26–IS903–ISCR1 was detected in 11 isolates being, to our knowledge, the first study that describes this genetic complex. Due to the wide variability observed, no relationship was determined among these mobile genetic elements and β-lactam resistance. More investigations regarding the genetic composition of these elements are needed to understand the role of multiple types of integrons and insertion sequences on the dissemination of antimicrobial resistance genes among different environments

Surveillance of listeriosis in Navarre, Spain, 1995-2005--epidemiological patterns and characterisation of clinical and food isolates

We monitored the incidence of human listeriosis in Navarre, a region in north of Spain between 1995 and 2005, and carried out the characterisation of Listeria monocytogenes isolates obtained from clinical samples and ready-to-eat products (sliced cooked meat, smoked salmon and liver pate). The active surveillance requesting hospitals to notify all listeriosis cases (n=40) yielded higher incidence rates (average annual rate 0.65/100,000 inhabitants, range 0.18-1.18/100,000 inhabitants) than expected. Pregnant women were the largest group affected (n=13, 32.5% of the cases), with a peak in incidence during the last three years of the study period. From the 40 human cases we obtained 33 Listeria isolates. Serological and molecular characterisation by PFGE identified 20 different pulsotypes, which on three occasions enabled us to link sporadic cases into clusters. Although we could not identify the incriminated food product we found two clinical pulsotypes among smoked salmon and cooked meat isolates. Surveillance of listeriosis in Spain should be improved and coordinated with other European Union Member States in order to better estimate the burden of disease and to prevent foodborne outbreaks

Food safety and the contract catering companies: food handlers, facilities and HACCP evaluation

The evaluation of implementing the HACCP system in contract catering companies and assessment of the knowledge, attitudes and practices of the food handlers were the main objectives of this study. It was conducted in 20 companies throughout Spain, with visual inspection of facilities and food handler activities, as well as the distribution of a self-administrable questionnaire among the 105 participant catering workers. In addition, samples of surfaces and finished dishes were taken from each kitchen for microbiological evaluation. The results obtained from the questionnaires reflect extensive knowledge and correct behaviors among respondents with a middle- or highschool educational level, stability in the same workplace and positions of greater responsibility. However, the observation of hygiene practices revealed systematic incorrect procedures in 60% of the kitchens. Most observed deviations were related to the lack of HACCP training and information, the temperature of finished dishes, the storage areas and proper cleaning and disinfection. In summary, this study confirms the difficulties inherent in catering companies to effectively implement the HACCP system, such as the lack of well-trained personnel, lack of motivation or adhesion to said system on the part of the workers, and the lack of financial and economic resources to address the deficiencies in the facilities. Specific educational programs and grants are need for an adequate implementation of HACCP in this sector

In vitro mutagenicity assessment of fried meat-based food from mass catering companies

The current article aimed to evaluate the in vitro mutagenicity of ten fried meat-based food extracts obtained from different catering companies from Navarra (Spain). A miniaturized 6-well version of the Ames test in Salmonella typhimurium TA98, and the in vitro micronucleus test (OECD TG 487) in TK6 cells were performed. None of the ten extracts of fried meat-based food induced gene mutations in S. typhimurium TA98 with or without metabolic activation, but five induced chromosomal aberrations after 24 h treatment of TK6 without metabolic activation. More studies are needed to check the biological relevance of these in vitro studies

Catering services and HACCP: temperature assessment and surface hygiene control before and after audits and a specific training session

Proper application of HACCP in catering services involves monitoring decisive critical points. The purpose of this study was to assess food temperatures and surface hygiene control in two catering services in Navarra (Spain) at two different time periods: the first one after implementation of the HACCP system and the second period, after the initial supervision through audits and a specific training session regarding temperatures of products and hygienic conditions of surfaces and equipment because the majority of detected nonconformities were related to these parameters. The recorded temperatures of 650 cooked food products within the first period showed that only 65.1% of the hot dishes had a temperature higher than 65 °C, in accordance with Spanish legislation, and 12.9% of them showed a risky holding temperature (26%) compared to utensils for distribution (12.0%). As the impact of dirty surfaces on the hygienic quality of a finished product will depend on which step was being taken during dish elaboration when equipment or utensil was used, it is suggested that more restrictive limits be established regarding utensils and equipment that are in direct contact with the finished product (≤1 CFU/cm2). Results of the study demonstrate that a specific training session on these items has improved the temperature control of prepared meals and the effectiveness of cleaning and disinfection, essentials for guaranteeing the hygienic quality of prepared foods

Practices of deep-frying processes among food handlers in social food services in Navarra, Spain

Deep frying is one of the most used worldwide methods in food preparation, but controlling the oil quality (temperature and formation of polar compounds) is crucial. The main objective of this work was to assess the practices of food handlers with regard to the frying processes in social food services located in Navarra (a region of northern Spain). The study was performed in two phases: in the first one, a self-administrable questionnaire regarding the usual practices on food deep-frying processes was sent to the food services through the main social catering companies of Navarra participating in the study. In the second one, in situ monitoring of the frying practices was performed as verification tools of frying practices reported by food services and to check the oil quality. Almost half of the fryers exceeded the maximum recommended temperature to avoid the formation of toxic compounds (175 ◦C). Despite only one the fryers exceeded the maximum limit of polar compounds established by current Spanish regulation, the obtained values indicated that the oil had begun to degrade in 20% of the fryers. Oil temperature is an important factor that affects the quality of the oil. In addition, significant differences were found between the different frequencies of change or types of oils. We have noticed a lack of knowledge in relation to the risks associated to the bad management of frying oil. Therefore, it would be desirable to improve food handlers training in relation to this matter. Defining a periodic frequency of oil change according to its use and periodic controls of temperature and polar compounds (as part of the Hazard Analysis and Critical Control Point system), could be adequate tools to improve management of frying oil in food services