Salmonella infection in healthy pet reptiles: Bacteriological isolation and study of some pathogenic characters

The fecal samples from 213 captive reptiles were examined, and 29 (13.61%) Salmonella enterica isolates were detected: 14/62 (22.58%) from chelonians, 14/135 (10.37%) from saurians, and 1/16 (6.25%) from ophidians. The isolates were distributed among 14 different serotypes: Miami, Ebrie, Hermannsweder, Tiergarten, Tornov, Pomona, Poona, Goteborg, Abaetetube, Nyanza, Kumasi, Typhimurium, 50:b:z6, 9,12:z29:1,5, and a non-motile serotype with antigenic formula 1,4,[5],12:-:-. Salmonella typhimurium and 50:b:z6 isolates showed the spv plasmid virulence genes, responsible of the capability to induce extra-intestinal infections. In some cases, pulsed field gel electrophoresis revealed different profiles for the strains of the same serotypes, showing different origins, whereas a common source of infection was supposed when one pulsotype had been observed for isolates of a serovar. Twenty-seven (93.10%) isolates showed resistance to one or more antibiotics. Ceftazidime was active to all the tested isolates, whereas the highest percentages of strains were no susceptible to tigecycline (93.10%), streptomycin (89.66%), and sulfonamide (86.21%)

Genome-wide identification of host-segregating epidemiological markers for source attribution in <i>Campylobacter jejuni</i>

Campylobacter is among the most common worldwide causes of bacterial gastroenteritis. This organism is part of the commensal microbiota of numerous host species, including livestock, and these animals constitute potential sources of human infection. Molecular typing approaches, especially multi-locus sequence typing (MLST), have been used to attribute the source of human campylobacteriosis by quantifying the relative abundance of alleles, at 7 MLST loci, among isolates from animal reservoirs and human infection, implicating chicken as a major infection source. The increasing availability of bacterial genomes provides data on allelic variation at loci across the genome, providing the potential to improve the discriminatory power of data for source attribution. Here we present a source attribution approach based on the identification of novel epidemiological markers among a reference pan-genome list of 1810 genes identified through gene-by-gene comparison of 884 genomes of C. jejuni isolates from animal reservoirs, the environment and clinical cases. Fifteen loci, involved in metabolic activities, protein modification, signal transduction and stress response, or coding for hypothetical proteins, were selected as host-segregating markers and used to attribute the source of 42 French and 281 UK clinical C. jejuni isolates. Consistent with previous studies of British campylobacteriosis, analyses performed using STRUCTURE software, attributed 56.8% of British clinical cases to chicken, emphasizing the importance of this host reservoir as an infection source in the UK. However, among French clinical isolates, approximately equal proportions of isolates were attributed to chicken and ruminant reservoirs suggesting possible differences in the relative importance of animal host reservoirs and indicating a benefit for further national-scale attribution modelling to account for differences in production, behaviour and food consumption.IMPORTANCE: Accurately quantifying the relative contribution of different host reservoirs to human Campylobacter infection is an ongoing challenge. This study based on the development of a novel source attribution approach, provides the first results of source attribution in Campylobacter jejuni in France. A systematic analysis using gene-by-gene comparison of 884 genomes of C. jejuni isolates, with a pan-genome list of genes, identified 15 novel epidemiological markers for source attribution. The different proportions of French and UK clinical isolates attributed to each host reservoir illustrates a potential role for local/national variations in C. jejuni transmission dynamics.</p

Heterogeneity of Persistence of Salmonella enterica Serotype Senftenberg Strains Could Explain the Emergence of this Serotype in Poultry Flocks

Salmonella enterica serotype Senftenberg (S. Senftenberg) has recently become more frequent in poultry flocks. Moreover some strains have been implicated in severe clinical cases. To explain the causes of this emergence in farm animals, 134 S. Senftenberg isolates from hatcheries, poultry farms and human clinical cases were analyzed. Persistent and non-persistent strains were identified in chicks. The non-persistent strains disappeared from ceca a few weeks post inoculation. This lack of persistence could be related to the disappearance of this serotype from poultry farms in the past. In contrast, persistent S. Senftenberg strains induced an intestinal asymptomatic carrier state in chicks similar to S. Enteritidis, but a weaker systemic infection than S. Enteritidis in chicks and mice. An in vitro analysis showed that the low infectivity of S. Senftenberg is in part related to its low capacity to invade enterocytes and thus to translocate the intestinal barrier. The higher capacity of persistent than non-persistent strains to colonize and persist in the ceca of chickens could explain the increased persistence of S. Senftenberg in poultry flocks. This trait might thus present a human health risk as these bacteria could be present in animals before slaughter and during food processing

The public health risk posed by Listeria monocytogenes in frozen fruit and vegetables including herbs, blanched during processing

A multi-country outbreak ofListeria monocytogenesST6 linked to blanched frozen vegetables (bfV)took place in the EU (2015–2018). Evidence of food-borne outbreaks shows thatL. monocytogenesisthe most relevant pathogen associated with bfV. The probability of illness per serving of uncooked bfV,for the elderly (65–74 years old) population, is up to 3,600 times greater than cooked bfV and verylikely lower than any of the evaluated ready-to-eat food categories. The main factors affectingcontamination and growth ofL. monocytogenesin bfV during processing are the hygiene of the rawmaterials and process water; the hygienic conditions of the food processing environment (FPE); andthe time/Temperature (t/T) combinations used for storage and processing (e.g. blanching, cooling).Relevant factors after processing are the intrinsic characteristics of the bfV, the t/T combinations usedfor thawing and storage and subsequent cooking conditions, unless eaten uncooked. Analysis of thepossible control options suggests that application of a complete HACCP plan is either not possible orwould not further enhance food safety. Instead, specific prerequisite programmes (PRP) andoperational PRP activities should be applied such as cleaning and disinfection of the FPE, water control,t/T control and product information and consumer awareness. The occurrence of low levels ofL. monocytogenesat the end of the production process (e.g.<10 CFU/g) would be compatible with thelimit of 100 CFU/g at the moment of consumption if any labelling recommendations are strictly followed(i.e. 24 h at 5°C). Under reasonably foreseeable conditions of use (i.e. 48 h at 12°C),L. monocytogeneslevels need to be considerably lower (not detected in 25 g). Routine monitoring programmes forL. monocytogenesshould be designed following a risk-based approach and regularly revised based ontrend analysis, being FPE monitoring a key activity in the frozen vegetable industry

Evaluation of the application for new alternative biodiesel production process for rendered fat including Category 1 animal by‐products (BDI‐RepCat® process, AT)

[EN] A new alternative method for the production of biodiesel from rendered fat, including animal by-product (ABP) Category 1 tallow, was evaluated. The method consists of a conversion phase, based on esterification and transesterification in a single step (at temperature ≥ 200°C, pressure ≥ 70 bar with a retention time ≥ 15 min), using MgO as a catalyst and in the presence of methanol (10–15%), followed by vacuum distillation (at ≥ 150°C, ≤ 10 mbar) of the end-product, biodiesel and the co-product, glycerine. Prions (PrPSc), which are abnormal isoforms of the prion protein, were considered by the applicant to be the most resistant hazard. In accordance with previous EFSA Opinions and current expert evaluation, a reduction in prion infectivity, or detectable PrPSc, of at least 6 log10 should be achieved for the process to be considered equivalent to the processing method laid down in the Regulation (EU) No 142/2011. Published data from an experimental replication of the conversion step of the biodiesel production process under consideration were provided, which showed an at least 6 log10 reduction in detectable PrPSc, by Western blot, in tallow that had been spiked with murine and human prion strains. In addition, it was demonstrated that the presence of methanol does not affect the recovery or detection of PrPSc from a biodiesel substrate. Based on scientific literature, the vacuum distillation step has been shown to be capable of achieving an additional 3 log10 reduction in PrPSc. Therefore, the proposed alternative method is considered to be at least equivalent to the processing method laid down in the legislation for the production of biodiesel from raw materials including Category 1 ABPSIThe Panel wishes to thank Katrin Bote for the support provided to this scientific outpu

Microbiological safety of aged meat

The impact of dry-ageing of beef and wet-ageing of beef, pork and lamb on microbiological hazards and spoilage bacteria was examined and current practices are described. As ‘standard fresh’ and wet-aged meat use similar processes these were differentiated based on duration. In addition to a description of the different stages, data were collated on key parameters (time, temperature, pH and aw) using a literature survey and questionnaires. The microbiological hazards that may be present in all aged meats included Shiga toxin-producing Escherichia coli (STEC), Salmonella spp., Staphylococcus aureus, Listeria monocytogenes, enterotoxigenic Yersinia spp., Campylobacter spp. and Clostridium spp. Moulds, such as Aspergillus spp. and Penicillium spp., may produce mycotoxins when conditions are favourable but may be prevented by ensuring a meat surface temperature of −0.5 to 3.0°C, with a relative humidity (RH) of 75–85% and an airflow of 0.2–0.5 m/s for up to 35 days. The main meat spoilage bacteria include Pseudomonas spp., Lactobacillus spp. Enterococcus spp., Weissella spp., Brochothrix spp., Leuconostoc spp., Lactobacillus spp., Shewanella spp. and Clostridium spp. Under current practices, the ageing of meat may have an impact on the load of microbiological hazards and spoilage bacteria as compared to standard fresh meat preparation. Ageing under defined and controlled conditions can achieve the same or lower loads of microbiological hazards and spoilage bacteria than the variable log10 increases predicted during standard fresh meat preparation. An approach was used to establish the conditions of time and temperature that would achieve similar or lower levels of L. monocytogenes and Yersinia enterocolitica (pork only) and lactic acid bacteria (representing spoilage bacteria) as compared to standard fresh meat. Finally, additional control activities were identified that would further assure the microbial safety of dry-aged beef, based on recommended best practice and the outputs of the equivalence assessment.info:eu-repo/semantics/publishedVersio

Evaluation of a multi-step catalytic co-processing hydrotreatment for the production of renewable fuels using Category 3 animal fat and used cooking oils

An alternative method for the production of renewable fuels from rendered animal fats (pretreated using methods 1–5 or method 7 as described in Annex IV of Commission Regulation (EC) No 2011/142) and used cooking oils, derived from Category 3 animal by-products, was assessed. The method is based on a catalytic co-processing hydrotreatment using a middle distillate followed by a stripping step. The materials must be submitted to a pressure of at least 60 bars and a temperature of at least 270°C for at least 4.7 min. The application focuses on the demonstration of the level of reduction of spores from non-pathogenic spore-forming indicator bacterial species (Bacillus subtilis and Desulfotomaculum kuznetsovii), based on a non-systematic review of published data and additional extrapolation analyses. The EFSA BIOHAZ Panel considers that the application and supporting literature contain sufficient evidence that the proposed alternative method can achieve a reduction of at least 5 log10 in the spores of B. subtilis and a 12 log10 reduction in the spores of C. botulinum. The alternative method under evaluation is considered at least equivalent to the processing methods currently approved in the Commission Regulation (EU) No 2011/142.info:eu-repo/semantics/publishedVersio

The use of the so-called ‘superchilling’ technique for the transport of fresh fishery products

Superchilling entails lowering the fish temperature to between the initial freezing point of the fish and about 1–2°C lower. The temperature of superchilled fresh fishery products (SFFP) in boxes without ice was compared to that of products subject to the currently authorised practice in boxes with ice (CFFP) under the same conditions of on-land storage and/or transport. A heat transfer model was developed and made available as a tool to identify under which initial configurations of SFFP the fish temperature, at any time of storage/transport, is lower or equal to CFFP. A minimum degree of superchilling, corresponding to an ice fraction in the fish matrix of SFFP equal or higher than the proportion of ice added per mass of fish in CFFP, will ensure with 99–100% certainty (almost certain) that the fish temperature of SFFP and the consequent increase of relevant hazards will be lower or equal to that of CFFP. In practice, the degree of superchilling can be estimated using the fish temperature after superchilling and its initial freezing point, which are subject to uncertainties. The tool can be used as part of ‘safety-by-design’ approach, with the reliability of its outcome being dependent on the accuracy of the input data. An evaluation of methods capable of detecting whether a previously frozen fish is commercially presented as ‘superchilled’ was carried out based on, amongst others, their applicability for different fish species, ability to differentiate fresh fish from fish frozen at different temperatures, use as a stand-alone method, ease of use and classification performance. The methods that were considered ‘fit for purpose’ are Hydroxyacyl-coenzyme A dehydrogenase (HADH) test, α-glucosidase test, histology, ultraviolet–visible–near–infrared (UV-VIS/NIR) spectroscopy and hyperspectral imaging. These methods would benefit from standardisation, including the establishment of threshold values or classification algorithms to provide a practical routine test.info:eu-repo/semantics/publishedVersio