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Electrical stimulation applied to individual organs, peripheral nerves, or specific brain regions has been used to treat a range of medical conditions. In cardiovascular disease, autonomic dysfunction contributes to the disease progression and electrical stimulation of the vagus nerve has been pursued as a treatment for the purpose of restoring the autonomic balance. However, this approach lacks selectivity in activating function- and organ-specific vagal fibers and, despite promising results of many preclinical studies, has so far failed to translate into a clinical treatment of cardiovascular disease. Here we report a successful application of optogenetics for selective stimulation of vagal efferent activity in a large animal model (sheep). Twelve weeks after viral transduction of a subset of vagal motoneurons, strong axonal membrane expression of the excitatory light-sensitive ion channel ChIEF was achieved in the efferent projections innervating thoracic organs and reaching beyond the level of the diaphragm. Blue laser or LED light (>10 mW mm−2; 1 ms pulses) applied to the cervical vagus triggered precisely timed, strong bursts of efferent activity with evoked action potentials propagating at speeds of ∼6 m s−1. These findings demonstrate that in species with a large, multi-fascicled vagus nerve, it is possible to stimulate a specific sub-population of efferent fibers using light at a site remote from the vector delivery, marking an important step towards eventual clinical use of optogenetic technology for autonomic neuromodulation. • Described is a method of selective efferent vagus nerve stimulation using light. • Vagal preganglionic neurons are targeted to express light-sensitive channels. • Specific efferent VNS by light delivery to the cervical vagus is achieved in a large animal model. • Demonstrates feasibility of using optogenetic technology for autonomic neuromodulation. [ABSTRACT FROM AUTHOR]

Listeria monocytogenes is a potential hazard for food safety and therefore for public health. The aim of the present study was to determine the prevalence and characteristics of L. monocytogenes in Polish ready-to-eat (RTE) meat products for retail sale. Among the 184,439 food samples collected within the framework of a national official control and monitoring program, only 0.3% were positive for L. monocytogenes. A significant group of products that did not meet the criteria were RTE meat products. This group accounted for 40% of all noncompliant samples. Seventy L. monocytogenes isolates from these RTE meat products (meat, sausages, and delicatessen products with meat) were examined. The majority of the tested isolates (51%) belonged to serogroup 1/2a-3a followed by 1/2c-3c (21%), 1/2b-3b-7 (14%), and 4ab-4b-4d-4e (13%). Serogroup 4a-4c was not present among the tested isolates. All L. monocytogenes isolates harbored the virulence-associated genes inlA, inlC, inlJ, and lmo2672. The llsX marker was detected in 12 (17%) of the 70 isolates. Ampicillin resistance was the most common resistance phenotype and was identified in 83% of the L. monocytogenes isolates. A low incidence of resistance to amoxicillin-clavulanic acid (6% of isolates) was also detected. All L. monocytogenes isolates were susceptible to chloramphenicol, gentamicin, ciprofloxacin, meropenem, sulfamethoxazole-trimethoprim, tetracycline, and erythromycin. This work provides useful information regarding contamination of RTE meat products with L. monocytogenes, which may have implications for food safety risks. [ABSTRACT FROM AUTHOR]

Abstract The sixty-seven nonpathogenic Listeria spp. strains isolated from food and food processing environments in Poland were examined for the presence of benzalkonium chloride (BC) resistance cassette (bcrABC) and four different variants of cadmium resistance determinants (cadA1-cadA4). All the strains were phenotypically resistant to cadmium and 22 among them were also resistant to BC. PCR-based analysis revealed that bcrABC cassette was harbored by 95.5% of the strains phenotypically resistant to BC. All of them harbored also either cadA1 or cadA2 genes (none carried cadA3 or cadA4), which corresponded to the presence of plasmids with two restriction patterns. The strains resistant to cadmium but susceptible to BC harbored only the cadA1 gene variant. DNA-DNA hybridization analysis showed that all the identified bcrABC , cadA 1 and cadA2 genes were located within plasmids, classified into 11 groups of RFLP profiles. Only one of the plasmids – pLIS1 of Listeria welshimeri (carrying bcrABC and cadA2) – was capable of efficient conjugal transfer from nonpathogenic Listeria isolates to a pathogenic Listeria monocytogenes strain. Analysis of the complete nucleotide sequence of pLIS1 (the first sequenced plasmid of L. welshimeri species) revealed the presence of genes involved in plasmid replication, stabilization and transfer as well as genes conferring resistance phenotypes. Comparative analysis showed that pLIS1 genome is highly similar to a group of plasmids originating from L. monocytogenes strains. A common feature of pLIS1 and its relatives, besides the presence of the resistance genes, is the presence of numerous transposable elements (TEs). The analysis revealed the important role of TEs in both promoting genetic rearrangements within Listeria spp. plasmids and the acquisition of resistance determinants. Highlights • Nonpathogenic cadmium resistant Listeria spp. carry either cadA1 or cadA2 genes • Majority of strains resistant to benzalkonium chloride carry bcrABC genes • The resistance determinants are located within plasmids of 11 RFLP patterns • Plasmid pLIS1 of L. welshimeri is capable of efficient conjugal transfer • pLIS1 contains bcrABC and cadA2 as well as numerous transposable elements [ABSTRACT FROM AUTHOR]

The aim of this study was to investigate the influence of the slaughter technique (Halal vs Classical slaughter) on the superficial contamination of cattle carcasses, by using traditional microbiological procedures and 16S rDNA metagenetics. The purpose was also to investigate the neck area to identify bacteria originating from the digestive or the respiratory tract. Twenty bovine carcasses (10 from each group) were swabbed at the slaughterhouse, where both slaughtering methods are practiced. Two swabbing areas were chosen: one “legal” zone of 1600 cm 2 (composed of zones from rump, flank, brisket and forelimb) and locally on the neck area (200 cm 2 ). Samples were submitted to classical microbiology for aerobic Total Viable Counts (TVC) at 30 °C and Enterobacteriaceae counts, while metagenetic analysis was performed on the same samples. The classical microbiological results revealed no significant differences between both slaughtering practices; with values between 3.95 and 4.87 log CFU/100 cm 2 and 0.49 and 1.94 log CFU/100 cm 2 , for TVC and Enterobacteriaceae respectively. Analysis of pyrosequencing data showed that differences in the bacterial population abundance between slaughtering methods were mainly observed in the “legal” swabbing zone compared to the neck area. Bacterial genera belonging to the Actinobacteria phylum were more abundant in the “legal” swabbing zone in “Halal” samples, while Brevibacterium and Corynebacterium were encountered more in “Halal” samples, in all swabbing areas. This was also the case for Firmicutes bacterial populations (families of Aerococcaceae , Planococcaceae ). Except for Planococcoceae , the analysis of Operational Taxonomic Unit (OTU) abundances of bacteria from the digestive or respiratory tract revealed no differences between groups. In conclusion, the slaughtering method does not influence the superficial microbiological pattern in terms of specific microbiological markers of the digestive or respiratory tract. However, precise analysis of taxonomy at the genus level taxonomy highlights differences between swabbing areas. Although not clearly proven in this study, differences in hygiene practices used during both slaughtering protocols could explain the differences in contamination between carcasses from both slaughtering groups. [ABSTRACT FROM AUTHOR]

A total of 127 Listeria isolates from food and food processing environments, including 75 L. innocua , 49 L. welshimeri , 2 L. seeligeri and 1 L. grayi were tested for susceptibility to eight antimicrobials, benzalkonium chloride (BC), cadmium and arsenic. The isolates were also screened for the presence of extrachromosomal genetic elements - plasmids, and their restriction pattern types were determined. All strains were susceptible to ampicillin, ciprofloxacin, erythromycin, gentamicin, rifampicin, trimethoprim and vancomycin. Two of the L. innocua isolates showed resistance to tetracycline and minocycline. The resistance was determined by the presence of chromosomal localization of tet (M) gene, which was not integrated in the transposon Tn 916 -Tn 1545 family. Of analyzed isolates, 18.11% and 55.91% isolates were resistant to BC and cadmium, respectively, but all were susceptible to arsenic. Resistance to BC was correlated with resistance to cadmium - all BC resistant isolates were also resistant to cadmium. On the other hand, 67.61% of cadmium-resistant isolates were susceptible to BC, suggesting that cadmium and BC resistance were not always concurrent in Listeria species. 48.03% of isolates contained plasmids. The size of most of the identified replicons was in the range of 50–90 kb. All plasmids were classified into 12 groups with identical restriction pattern (I–XII). Interestingly, plasmids belonging to the same group were determined in isolates of the same species. Only in one case, plasmids with I-type profile were identified in L. innocua and L. welshimeri. There was an association between resistance to BC and plasmid DNA presence: all resistant isolates carried a plasmid. A correlation between resistance to cadmium and plasmid carriage was also observed in L. innocua and L. seeligeri isolates, but among resistant L. welshimeri, 23.08% of isolates did not have plasmids. This may suggest that resistance is associated with determinants located within the chromosome. To elucidate the adaptation strategies and ecology of Listeria spp., it is important to have a better understanding of its resistance to antimicrobials and environmental toxicants such as heavy metals and disinfectants. [ABSTRACT FROM AUTHOR]

Steak tartare is a popular meat dish in Belgium. It is prepared with raw minced beef and is eaten with sauce, vegetables, and spices. Because it contains raw meat, steak tartare is highly prone to bacterial spoilage. The objective of this study was to explore the diversity of bacterial flora in steak tartare in Belgium according to the source and to determine which bacteria are able to grow during shelf life. A total of 58 samples from butchers' shops, restaurants, sandwich shops, and supermarkets were collected. These samples were analyzed using 16S rDNA metagenetics, a classical microbiological technique, and quantitative real-time PCR (qPCR) targeting the Lactobacillus genus. Samples were analyzed at the beginning and at the end of their shelf life, except for those from restaurants and sandwich shops, which were analyzed only on the purchase date. Metagenetic analysis identified up to 180 bacteiial species and 90 genera in some samples. But only seven bacterial species were predominant in the samples, depending on the source: Brochothrix thermosphacta, Lactobacillus algidus, Lactococcus piscium, Leuconostoc gelidum, Photobacterium kishitani, Pseudomonas spp., and Xanthomonas oryzae. With this work, an alternative method is proposed to evaluate the total flora in food samples based on the number of reads from metagenetic analysis and the results of qPCR. The degree ot underestimation of aerobic plate counts at 30°C estimated with the classical microbiology method was demonstrated in comparison with the proposed culture-independent method. Compared with culture-based methods, metagenetic analysis combined with qPCR targeting Lactobacillus provides valuable information for characterizing the bacterial flora of raw meat. [ABSTRACT FROM AUTHOR]

Milk kefir is produced by fermenting milk in the presence of kefir grains. This beverage has several benefits for human health. The aim of this experiment was to analyze 5 kefir grains (and their products) using a targeted metagenetic approach. Of the 5 kefir grains analyzed, 1 was purchased in a supermarket, 2 were provided by the Ministry of Agriculture (Namur, Belgium), and 2 were provided by individuals. The metagenetic approach targeted the V1-V3 fragment of the 16S ribosomal (r)DNA for the grains and the resulting beverages at 2 levels of grain incorporation (5 and 10%) to identify the bacterial species population. In contrast, the 26S rDNA pyrosequencing was performed only on kefir grains with the aim of assessing the yeast populations. In parallel, pH measurements were performed on the kefir obtained from the kefir grains using 2 incorporation rates. Regarding the bacterial population, 16S pyrosequencing revealed the presence of 20 main bacterial species, with a dominance of the following: Lactobacillus kefiranofaciens, Lactococcus lactis ssp. cremoris, Gluconobacter frateurii, Lactobacillus kefiri, Acetobacter orientalis, and Acetobacter lovaniensis. An important difference was noticed between the kefir samples: kefir grain purchased from a supermarket (sample E) harbored a much higher proportion of several operational taxonomic units of Lactococcus lactis and Leuconostoc mesenteroides. This sample of grain was macroscopically different from the others in terms of size, apparent cohesion of the grains, structure, and texture, probably associated with a lower level of Lactobacillus kefiranofaciens. The kefir (at an incorporation rate of 5%) produced from this sample of grain was characterized by a lower pH value (4.5) than the others. The other 4 samples of kefir (5%) had pH values above 5. Comparing the kefir grain and the kefir, an increase in the population of Gluconobacter in grain sample B was observed. This was also the case for Acetobacter orientalis in sample D. In relation to 26S pyrosequencing, our study revealed the presence of 3 main yeast species: Naumovozymaspp., Kluyveromyces marxianus, and Kazachastania khefir. For Naumovozyma, further studies are needed to assess the isolation of new species. In conclusion, this study has proved that it is possible to establish the patterns of bacterial and yeast composition of kefir and kefir grain. This was only achieved with the use of high-throughput sequencing techniques. [ABSTRACT FROM AUTHOR]

The purpose of the present study was to determine the prevalence of thermophilic Campylobacter in poultry, pork, and beef meat at the retail level and to identify the main categories of meat representing the most significant reservoirs of Campylobacter. A monitoring study was conducted throughout Poland from 2009 to 2013. A total of 1,700 fresh meat samples were collected from supermarkets, large retail outlets, and smaller stores. Thermophilic Campylobacter species were detected in 690 (49.3%) of 1,400 poultry samples collected from retail trade. Strains were isolated from 50.2 and 41.1% of raw chicken and turkey meat samples, respectively, and from 50.1 and 42.6% of raw chicken and turkey giblets. The incidence of Campylobacter spp. on pork ( 10.6%) and beef ( 10.1 %) was significantly lower than on poultry. Campylobacter jejuni was the most prevalent Campylobacter species in chicken (46.6%), pork (68.6%), and beef (66.7%), and Campylobacter coli was the most frequently isolated Campylobacter species in turkey meat (71.2%). This study revealed that retail raw meats are often contaminated with Campylobacter; however, the prevalence of these pathogens is markedly different in different meats. Raw retail meats are potential vehicles for transmitting foodborne diseases, and our findings stress the need for increased implementation of hazard analysis critical control point programs and consumer food safety education efforts. [ABSTRACT FROM AUTHOR]

Abstract: Seafood can pose a public health concern to consumers. It is often consumed raw and may be contaminated with several foodborne pathogens. In order to guarantee the safety of seafood, real-time polymerase chain reaction (PCR) protocols may be used as these enable results to be provided within 24 h. The first goal of our work was to develop real-time PCR protocols enabling the detection of six foodborne pathogens that may be present in seafood products (Campylobacter jejuni, Campylobacter coli, enterohemorrhagic Escherichia coli, Salmonella spp., Vibrio parahaemolyticus, and Vibrio vulnificus). The corresponding gene targets were: 50S/VS1, rfbE, ttr, tlh, and vvp. A multiplex PCR was also developed to detect the virulence genes of V. parahaemolyticus: tdh and trh. A total of 420 bacterial strains belonging to four different genera/strains were used in this study. Sensitivity and specificity were always 100%, except in the case of Salmonella spp., where three strains were not detected by our PCR protocols. The second objective of our work was to assess the detection limit of our real-time PCR protocols on artificially contaminated seafood products (raw shrimps, cooked shrimps, and raw mussels), purchased in public stores. Six different levels of contamination were assayed in four replicates for each matrix. The real-time PCR protocols enabled a better level of detection than the ISO methods, except for Salmonella in raw shrimps and for V. vulnificus in shrimps (raw and cooked). The estimated level of detection was between 1 and 47 cfu/25 g sample for the ISO norms and between 1 and 315 cfu/25 g sample for the real-time PCR protocols tailored in our work. The real-time PCRs developed in our work allowed for good selectivity, sensitivity, and specificity. The sensitivity on seafood products was estimated at a level of 100%, except for Salmonella (97%). In the spiking assays, the levels of detection were lower with the real-time PCR protocol than those obtained with the ISO method. This was not the case for V. vulnificus in raw and cooked shrimps and for Salmonella in raw shrimps. These real-time PCR protocols appear to be good alternative methods for surveillance of seafood products to ensure the absence of foodborne pathogens. One additional conclusion is that laboratories have to use enrichment media that are compatible with those recommended by ISO standards. This may facilitate the isolation of the pathogen if the real-time PCR protocol gives a suspect positive signal during the first step of the seafood analysis. [Copyright &y& Elsevier]

In 2005, the Belgian authorities reported a Listeria monocytogenes contamination episode in cheese made from raw goat's milk. The presence of an asymptomatic shedder goat in the herd caused this contamination. On the basis of data collected at the time of the episode, a retrospective study was performed using an exposure assessment model covering the production chain from the milking of goats up to delivery of cheese to the market. Predictive microbiology models were used to simulate the growth of L. monocytogenes during the cheese process in relation with temperature, pH, and water activity. The model showed significant growth of L. monocytogenes during chilling and storage of the milk collected the day before the cheese production (median increase of 2.2 log CFU/ml) and during the addition of starter and rennet to milk (median increase of 1.2 log CFU/ml). The L. monocytogenes concentration in the fresh unripened cheese was estimated to be 3.8 log CFU/g (median). This result is consistent with the number of L. monocytogenes in the fresh cheese (3.6 log CFU/g) reported during the cheese contamination episode. A variance-based method sensitivity analysis identified the most important factors impacting the cheese contamination, and a scenario analysis then evaluated several options for risk mitigation. Thus, by using quantitative microbial risk assessment tools, this study provides reliable information to identify and control critical steps in a local production chain of cheese made from raw goat's milk. [ABSTRACT FROM AUTHOR]

Abstract: A total of 471 Listeria monocytogenes isolates from different types of food and food-related sources in Poland during 2004–2010 were examined. This number includes 200 isolates from fish, 144 from fresh and frozen vegetables, 43 ready-to-eat products (deli foods, cold cuts), 13 from dairy products, 16 from raw meats, 15 from confectionery products and 40 directly from processing plants. All isolates were subjected to serotyping and lineage assays using PCR, and antimicrobial susceptibility using E-test and a broth microdilution method. Of all isolates, 256 (54.4%), 120 (25.5%), 59 (12.5%), 36 (7.6%) were identified as serotypes 1/2a (or 3a), 1/2c (or 3c), 1/2b (or 3b or 7), and 4b (or 4d or 4e), respectively. A direct correlation between the most common serotypes and three L. monocytogenes lineages was also observed. All L. monocytogenes isolates belonged to lineages I (20.2%) and II (79.8%). All strains were sensitive to ampicillin, amoxicillin, gentamicin, erythromycin, trimethoprim, rifampicin, vancomycin, chloramphenicol and sulfamethoxazol. Two of the L. monocytogenes strains (0.42%) showed phenotypic resistance. One strain was resistant to tetracycline and minocycline due to the presence of tet(M). It did not carry gene int, which may indicate that the tet(M) gene in this strain was not integrated in the transposon Tn916-Tn1545 family. The resistance of the second strain to ciprofloxacin and norfloxacin was attributed to active efflux associated with overexpression of gene lde. Our data indicate the low prevalence of antimicrobial resistance among L. monocytogenes isolates from food and food-related sources in Poland. [Copyright &y& Elsevier]

Abstract: Oxidative stress is widely recognized to contribute to neuronal death during various pathological conditions and ageing. In the enteric nervous system (ENS), reactive oxygen species have been implicated in the mechanism of age-associated neuronal loss. The neurotrophic factors, neurotrophin 3 (NT-3) and glial cell line-derived neurotrophic factor (GDNF), are important in the development of enteric neurons and continue to be expressed in the gut throughout life. It has therefore been suggested that they may have a neuroprotective role in the ENS. We investigated the potential of NT-3 and GDNF to prevent the death of enteric ganglion cells in dissociated cell culture after exposure to hydrogen peroxide (H2O2). H2O2 treatment resulted in a dose-dependent death of enteric neurons and glial cells, as demonstrated by MTS assay, bis-benzimide and propidium iodide staining and immunolabelling. Cultures treated with NT-3 prior to exposure showed reduced cell death compared to untreated control or GDNF-treated cultures. GDNF treatment did not affect neuronal survival in H2O2-treated cultures. These results suggest that NT-3 is able to enhance the survival of enteric ganglion cells exposed to oxidative stress. [Copyright &y& Elsevier]

Abstract: This study presents the results of investigations on the susceptibility of Campylobacter spp. strains isolated from chicken meat and giblets to fluorochinolones (ciprofloxacin), macrolides (erythromycin), tetracyclines (tetracycline) and aminoglycosides (gentamicin) andV an analysis of the molecular mechanisms of resistance to the selected antibiotics. Between January 2008 and December 2009 a total of 218 samples of chicken meat and giblets from retail trade in Poland were examined. Campylobacter bacteria were found in 143 samples, that is in 65.6% of the total number embraced by the study. Campylobacter coli was the most ubiquitous – its presence was determined in 108 samples out of 143 (75.5%), whereas Campylobacter jejuni was found in 35 of the contaminated samples (24.5%). The results obtained point to the high percentage (97.9%) of Campylobacter isolates resistant to ciprofloxacin. 92 strains (64.3%) were resistant to tetracycline, 14 (9.1%) to erythromycin and only 9 (6.3%) to gentamicin. Moreover, ten out of the 143 resistant Campylobacter strains (7.0%) were found to be resistant to at least three unrelated antibiotics. [Copyright &y& Elsevier]

Abstract: Several domestic varieties of kiwifruit including Actinidia deliciosa cv. ‘Hayward’, ‘Daeheung’, ‘Monty’, ‘Arimold’, ‘Jinmi’, ‘Hyangrok’ and A. chinensis cv. ‘Haenam’ and ‘Golden King’ were utilized for the processing of wines. Kiwifruit wines were analyzed for sugar content, alcohol, acidity, pH, color, total phenols and antioxidant activity by radical scavenging assay ABTS. The soluble solid content of the kiwifruit must ranged between 22 and 25% and then was fermented at 14°C during 4months. The amount of alcohol productions differed especially during the first week of fermentation. Among cultivars, ‘Haenam’ and ‘Arimold’ were relatively slower than other wines with regard to alcohol production rate. The yield of wine production was increased from 63.35% to 66.19% in weight by using processing with pectinase. The ‘L’ values of wine made from ‘Hayward’ and ‘Monty’ were lower and darker than those of wines prepared from other cultivars. Wine made from ‘Daeheung’ had total phenols of 790mg L−1, which was the highest among wines, followed by ‘Haenam’ and ‘Golden King’. ‘Daeheung’ also showed the highest antioxidant activity (22.55mM TE L−1), while ‘Arimold’ showed the lowest one (10.91mM TE L−1). The mean overall acceptability scores among kiwifruit wines ranged between 3.8 and 4.5. The sensory evaluations were higher in ‘Golden King’, ‘Monty’, ‘Daeheung’, ‘Arimold’, and ‘Hayward’ than those of ‘Haenam’, ‘Jinmi’, and ‘Hyangrok’. However, for commercial kiwifruit wine production not only overall sensory acceptance, but also functional properties such as total phenols and antioxidant activity, and fruit cultivation volume should be concerned. Therefore, three of kiwifruit cultivars which are ‘Golden King’, ‘Daeheung’, and ‘Hayward’ had the most desirable characteristics suitable for kiwifruit wine production. [Copyright &y& Elsevier]

In 2007 and 2008, a monitoring study was carried out in Poland to examine the occurrence of thermotolerant Campylobacter spp. in raw and cooked chicken products available on the retail market. A total of 912 samples were tested: 443 samples of raw chicken meat, 146 samples of giblets, and 323 ready-to-eat poultry products (150 samples of spit-roasted chicken, 56 samples of smoked chicken, and 117 samples of pâté and cold meats). A high level of contamination of raw chicken meat (51.7% o samples) and chicken giblets (47.3% of samples) was detected. However, thermotolerant Campylobacter spp. were found in only 1.2% of the ready-to-eat poultry products. [ABSTRACT FROM AUTHOR]

The risk of human salmonellosis through the consumption of minced pork meat in Belgium was assessed via a modular risk model covering pork meat production from lairage to human consumption. The main goal of the model was to give concrete options to reduce effectively the risk of human salmonellosis through the consumption of minced pork meat. These options (scenarios) were elaborated with reference to the international situation and the literature to give concrete and realistic possibilities for improving the microbiological quality of pork meat and to reduce the number of human salmonellosis cases per year in Belgium. The model estimates 15,376 cases of human salmonellosis per year in Belgium due to the consumption of minced pork meat. The results of the scenarios showed that the risk of human salmonellosis could be significantly reduced by efforts all along the pork meat production chain but also by efforts made by consumers. The responsibility of food business operators for the pork meat production chain is high in relation to the microbiological quality of meat delivery, especially at the slaughterhouse. Consumers also need to be aware of good hygiene practices during preparation of the meat at home. Cross-contamination with raw food can be avoided by changing the habits and the behavior of the household cook. The results of these scenarios would be useful for the food business operators involved in the pork meat chain and for public health authorities. [ABSTRACT FROM AUTHOR]

[Display omitted] • The concept of bioanalytical insight into the life of microbial populations is presented. • The flow analysis system operating as a real-time on-line monitor of biocatalytic activity is developed. • The developed MCFA monitor is useful for investigations on urease-positive bacteria growth. • Advantages of chemical monitoring and modelling of microbial growth are announced. In this publication an alternative approach to investigations of bacterial growth is proposed. Contrary to the conventional physical methods it is based on enzyme activity detection. The procedure for real-time and on-line monitoring of microbial ureolytic activity (applied as a model experimental biosystem) in the flow analysis format is presented. The developed fully-mechanized bioanalytical flow system is composed of solenoid micropumps and microvalves actuated by Arduino microcontroller. The photometric detection based on Nessler reaction is performed using dedicated flow-through optoelectronic detector made of paired light emitting diodes. The developed bioanalytical system allows discrete assaying of microbial urease in the wide range of activity up to 5.4 U mL−1 with detection limit below 0.44 U mL−1, a high sensitivity in the linear range of response (up to 200 mV U−1 mL and relatively high throughput (9 detection per hour). The proposed differential procedure of measurements (i.e. a difference between peaks register for sample with and without external addition of urea is treated as an analytical signal) allows elimination of interfering effects from substrate and products of biocatalysed reaction as well as other components of medium used for microbial growth. The developed bioanalytical system was successfully applied for the control of growth of urease-positive bacteria strains (Proteus vulgaris , Klebsiella pneumoniae and Paracoccus yeei) including examination of effects from various microbial cultivation conditions like temperature, composition of culture medium and amount of substrate required for induction of bacterial enzymatic activity. The developed bioanalytical flow system can be applied for metabolic activity-based estimation of parameters of lag and log phases of microbial growth as well as for detection of decline phase. [ABSTRACT FROM AUTHOR]

A survey was conducted to collect data on Salmonella prevalence, Escherichia coli counts (ECCs), and aerobic bacteria colony counts (ACCs) on pig carcasses after chilling at the 10 largest Belgian pig slaughterhouses during 2000 through 2004. Potential risk factors of contamination associated with production parameters, technical descriptions of the installations, and cleaning and disinfection methods were assessed during investigations in the slaughterhouses. These variables were used first in a univariate analysis and then were extended to a multivariate analysis with a logistic mixed regression model for Salmonella and a linear mixed model for ECCs and ACCs with slaughterhouses as the random effect. The results indicated high variability concerning Salmonella contamination among the 10 slaughterhouses, with prevalence ranging from 2.6 to 34.3% according to the area of origin. The median ECC and median ACC ranged from -0.43 to 1.11 log CFU/cm² and from 2.37 to 3.65 log CFU/cm², respectively. The results of the logistic and linear regressions revealed that some working practices such as scalding with steam, second flaming after polishing, and complete cleaning and disinfection of the splitting machine several times a day were beneficial for reducing Salmonella prevalence, ECCs, and ACCs. Changing the carcass hooks just before chilling, using water as the cleaning method, and a higher frequency of disinfection of the lairage seemed to be protective against E. coli in the multivariate mixed linear model. The monitoring of critical points, slaughterhouse equipment, good slaughtering practices, and effective washing and disinfection are the keys to obtaining good microbiological results. [ABSTRACT FROM AUTHOR]

Campylobacter-associated gastroenteritis remains an important cause of morbidity worldwide, and some evidence suggests that poultry is an important source of this foodborne infection in humans. This study was conducted to analyze the prevalence and genetic background of resistance of 149 Campylobacterjejuni and 54 Campylobacter coli strains isolated from broiler chicken carcasses and from stool samples of infected children in Poland from 2003 through 2005. Nearly all isolates were susceptible to macrolides and aminoglycosides. The highest resistance in both human and chicken strains was observed for ciprofloxacin (more than 40%), followed by ampicillin (13 to 21%), and tetracycline (8 to 29%). Resistance to ampicillin and tetracycline rose significantly between 2003 and 2005. Slight differences in resistance between human and chicken isolates indicate that although chicken meat is not the only source of Campylobacter infection in our population, it can be involved in the transmission of drug-resistant Campylobacter strains to humans. [ABSTRACT FROM AUTHOR]

Abstract: Relevance of a Salmonella serological detection technique was studied from complete results obtained from 9 pigs fattening units. Feces and overshoes were sampled at different periods after starting fattening (2, 3 and 4 months) while caecal contents were taken on the slaughter line. The bacteriological technique used was based on a Diasalm enrichment and a commercial test was used for serology on an average of ten animals per batch. The aim of this work was to establish a correlation between serological results obtained at slaughter (10 samples/batch) and bacteriological results. In this context, two types of logistic regression models were tested by considering alternatively serology and Salmonella detection in caecal contents as the dependent variables. Firstly, beside the fact that all logistic regression models show weak correlations, the first finding was that positive results in overshoes taken at 2 and 3 months are slightly correlated with serological status of herds (odds-ratios of 4.96 and 2.55). Secondly, when batches were characterized as positive on the basis of serological results, the probability of Salmonella recovery in caecal contents was higher than when the batches were considered as negative (odds-ratios comprised between 4.36 and 5.81). A major conclusion is that serology can be used to follow the improvement of an integrated pig production system, but is not the unique solution for assessing risk of Salmonella shedding from specific herds. [Copyright &y& Elsevier]

Abstract: Gold in nano-crystal size is known as highly active CO-oxidation catalyst. Using simple deposition techniques gold has been deposited as ∼3nm particles inside the channels of mesoporous silica–TiO2–MCM-48. In the presence of gold nano-particles the catalyst converts CO to CO2 at 50% level at −20°C. The composite is stable against sintering up to at least 200°C. XANES and EXAFS confirm the coexistence of elementary and ionic gold during the catalytic activity. [Copyright &y& Elsevier]

From 1997 to 1999, the prevalence of Salmonella was assessed at different stages through the pork, poultry, and beef meat production chains. Different dilutions of the initial sample suspension were analyzed to provide a semiquantitative evaluation of Salmonella contamination and to determine the most representative dilution necessary to detect a reduction in prevalence. An average of 300 samples for each type of meat were analyzed. According to Fisher's exact test, the dilution to be used to detect a reduction in prevalence was chosen based on an initial prevalence of 20 to 26%. Based on this introductory study, a new sampling plan representative of the nationwide Belgian meat production process was used from 2000 through to 2003. This study confirmed the consistently high rate and level of contamination of poultry meat: broiler and layer carcasses were the most contaminated samples followed by broiler fillets and poultry meat preparations. A constant and significant decrease in Salmonella prevalence was observed for pork carcasses, trimmings, and minced meat and for beef minced meat. Less than 3% of beef carcasses and trimming samples were positive for Salmonella. The Belgian plan, as utilized from 2000 to 2003, was suitable for monitoring of zoonoses because the sampling plan was representative of nationwide production processes, covered all periods of the year, and was executed by trained samplers and the analyses were carried out by recognized laboratories using an identical analytical method. [ABSTRACT FROM AUTHOR]

Performances of four detection methods were evaluated for recovery of Salmonella spp. in naturally contaminated fecal specimens of porcine origin. The NMKL 71 method consisted of enrichment in Rappaport-Vassiliadis broth and plating on xylose-lysine-desoxycholate medium, whereas the SP-VG-M002 method relied on a Diasalm enrichment followed by streaking on xylose-lysine-tergitol 4 agar (XLT-4). The VIDAS SLM method was composed of double enrichment in Muller-Kauffmann tetrathionate broth and in M broths before processing in a VIDAS device. If the results were positive, the VIDAS ICS immunoenrichment was performed and the result transferred onto three different selective media. The VIDAS ICS protocol is an immunoconcentration step followed by plating on XLT-4. Seventy-eight samples were tested with all four methods simultaneously, leading to 34 positive samples with at least one method. For this assay, VIDAS SLM revealed 31 positive samples (91.2%), whereas the average positive percentage of the three other methods was 37.3% (P < 0.001). Two-paired comparisons with the VIDAS SLM method were also performed. McNemar values were systematically highly significant (P < 0.001). The proportion of agreement was significantly inferior (P < 0.05) for the comparison of VIDAS ICS and VIDAS SLM (68.7%) compared with the two other paired comparisons (average percentage, 8 1.5%). The conclusion reached by this trial is that VIDAS SLM significantly improves the recovery of Salmonella in naturally contaminated fecal specimens. For the paired- comparisons, NMKL 71 and SP-VG-M002 were comparable in terms of efficiency, whereas the VIDAS ICS protocol, as established by the manufacturer for food samples only, seemed less efficient than the other two. [ABSTRACT FROM AUTHOR]

Neurones within the rostral ventromedial medulla project to the intermediolateral cell column of the spinal cord where they may influence sympathetic preganglionic neuronal activity controlling cutaneous vascular beds. Here we assess whether such neurones contribute to cutaneous sympathetic vasoconstrictor activity in a fever-like state induced by i.c.v. injection of E-series prostaglandin. In urethane-anaesthetised rats, we recorded population sympathetic activity to the tail (an index of vasoconstrictor discharge regulating cutaneous thermoregulatory circulations).A survey of the effects of GABA microinjections (200 mM; 60–80 nl; 111 sites in 57 rats) demonstrated that those into the rostral ventromedial medulla (in the region of raphe pallidus and magnus; approximately bregma −10 to −12 mm) markedly decreased (51%–100%) population sympathetic cutaneous vasoconstrictor activity during “normothermic control.” In contrast, injections at sites dorsal and lateral to this region tended to produce either a smaller decrease or have no effect. In heat-clamp (nine animals: body temperature 40–41 °C) cutaneous vasoconstrictor activity was decreased by 83±5%. I.c.v. prostaglandin E1 (100 ng and above) restored activity to, or above, control levels in these animals and where body temperature was maintained at control levels (12 animals: body temperature 35.5–36.5 °C). The depressant action of GABA was sustained in both conditions. GABA did not significantly influence concurrently recorded splanchnic nerve activity and heart rate in any condition although both were increased following i.c.v. prostaglandin E1 (500 ng).This study is the first to demonstrate that inhibition of neuronal activity within the rostral ventromedial medulla decreases sympathetic cutaneous vasoconstrictor activity during normothermic control and following i.c.v. prostaglandin E1 (both with and without heat-clamp). Therefore, sympathetic premotor neurones in this area contribute to vasoconstrictor drive in these conditions. In contrast, we were unable to demonstrate that the same area had a substantial involvement in the control of splanchnic nerve activity or heart rate, even when these were enhanced following i.c.v. prostaglandin E1. [Copyright &y& Elsevier]

This paper describes the monitoring of Salmonella in a closed pig production system in Belgium over a 2-year period. A sampling scheme including animal feeds and carcasses was designed to cover the entire chain of production from farrow to finishing pigs. Salmonella was detected by a method based on the use of semisolid Rappaport-Vassiliadis as a selective medium. The serotypes of the isolated strains were determined, and the antibiotic resistance of these strains to six antibiotics was also investigated. Feeds were found to be more contaminated than expected (10.2%, 34 of 332 samples). The percentage of positive fecal samples for pregnant sows (8.1%, 11 of 135 samples) was significantly higher than that for young and lactating sows (2.9%, 11 of 378 samples) (P < 0.05). The percentage of positive samples for colon contents collected at the slaughterhouse (47.3%, 88 of 186 samples) was significantly higher than that for feces collected during the fattening stage (5.6%, 18 of 320 samples) (P < 0.001). For carcass swab samples, the observed prevalence was 11.2% (17 of 152 samples). On farms, Salmonella recovery levels were higher for overshoe samples than for fecal samples, except for pregnant sows. Salmonella Typhimurium was the most frequently isolated serotype (32.2%, 55 of 171 samples), while Salmonella Brandenburg was predominant in the colon contents collected at the abattoir (21.4%, 18 of 84 samples). Feeds harbored a wide diversity of serotypes of minor epidemiological significance. Of 55 isolated strains of Salmonella Typhimurium, 11 (20%) were resistant to tetracycline, ampicillin, choramphenicol, streptomycin, trimethoprim, and nalidixic acid (R Type TeAmCSNa), while 12 (21.8%) were resistant to all of these antibiotics except nalidixic acid (R Type TeAmCS). The majority of Salmonella Typhimurium strains that exhibited resistance to more than four antimicrobial agents were characterized as Salmonella Typhimurium DT104 or as being closely related to... [ABSTRACT FROM AUTHOR]

Neuronal population discharges within the CNS and in somatic and sympathetic motor nerves often display oscillations. Peripheral oscillations may provide a window into central mechanisms, as they often show coherence with population activity of subsets of central neurones. The reduction in heat loss through the cutaneous circulation during fever may be mediated via sympathetic premotor neurones not utilised during normal temperature regulation. Consequently, here we assessed, in anaesthetised rats, whether the frequency signature of population sympathetic discharge observed in neurones innervating the tail (thermoregulatory) circulation changed during a fever-like response induced by intracerebroventricular injection of prostaglandin E1.We found that when core temperature was raised to 38.8–40.5°C sympathetic activity was abolished. Following administration of prostaglandin (400 ng or 1 μg per rat), activity was restored to levels seen prior to heating (154±53.5%; n=10). Injection of vehicle had no effect (n=7). Prior to heating when most animals were in central apnoea (14/18) two peaks were observed in autospectra of sympathetic activity: one at 0.68–0.93 Hz (T-peak) and another at the frequency of ventilation (2 Hz). Central respiratory drive was recruited during hyperthermia where it was 1:2 locked to the frequency of ventilation and following prostaglandin administration, an additional peak in sympathetic autospectra was seen at this frequency. Time-evolving spectra indicated that this peak resulted from the dynamic locking of the ‘T-peak’ to central respiratory drive.Our data show that during a fever-like response the dominant oscillations in sympathetic activity controlling a thermoregulatory circulation and their dynamic coupling to respiratory-related inputs are similar to those seen under normal conditions. Therefore, during this fever-like response, the neural substrate(s) underlying the oscillations is not reconfigured and remains capable of sculpturing the pattern of sympathetic neuronal discharge that may be regulated by several descending pathways. [Copyright &y& Elsevier]

The study describes the characterization of Listeria monocytogenes isolated from the general 2017–2019 national official control and monitoring sampling program. A total of 60,928 of ready-to-eat (RTE) food products were collected in retail in Poland, while the number of L. monocytogenes contaminated samples was 67 (0.1%). The majority of the strains belonged to molecular serotype IVb followed by IIa, frequently associated with human listeriosis. Furthermore, 61.2% of the isolates were resistant at least to one of the tested antimicrobials: penicillin, ampicillin, meropenem, erythromycin, sulfamethoxazole-trimethoprim, amoxicillin-clavulanic acid, ciprofloxacin, chloramphenicol, gentamicin, vancomycin, tetracycline and rifampicin. Virulence genes inlA , inlC , inlJ and lmo2672 were detected in all of the isolates. In our study the llsX gene (encoding LLS) exhibited 11.6% positivity. The 32 strains were grouped into 12 clonal complexes (CCs) which belong to the major clones that are in circulation in Europe. Among them, seven strains with the cgMLST close relatedness (CC2) were isolated from diverse food sectors, underlining a large circulation of this clone in Poland, most likely from multiple introduction sources. Additionally, two RTE strains CC6 and one CC37 were identified as closely related by cgMLST to two publicly available genomes of clinical strains isolated in Poland in 2012–2013. These results indicate the large strain circulation and point to RTE food products as a potential source of human listeriosis. The present study provided data to capture the contamination status of L. monocytogenes in foods at the retail level in Poland and assess the potential risk of this pathogen for human safety. • L. monocytogenes was isolated from 0.1% of the RTE food samples. • The most frequently identified molecular serogroups were IVb (51%) and IIa (42%). • 48% strains belonged to the major clonal complexes (CCs) circulating in Europe. • Seven strains (CC2) close relatedness were isolated from diverse food sectors. • Two strains CC6 and one C37 were closely related to clinical strains in Poland. [ABSTRACT FROM AUTHOR]

The aim of the present study is to investigate the prevalence and genetic diversity of Listeria monocytogenes in various fresh and frozen vegetable products available in Poland. The samples were collected at retail market within the framework of national official control and monitoring program. In the years 2016–2019 a total of 49 samples out of 8712 collected vegetable samples were positive for L. monocytogenes. Our findings demonstrated that the occurrence of L. monocytogenes in various vegetable products was generally low, on average only 0.56% in the studied years. All isolates were susceptible to 11 antimicrobial agents: penicillin, ampicillin, meropenem, erythromycin, sulfamethoxazole-trimethoprim, amoxicillin-clavulanic acid, ciprofloxacin, chloramphenicol, gentamicin, vancomycin, and tetracycline. All of them harbored virulence-associated genes (inlA , inlC , and lmo2672), 82% harbored inlJ gene and few of them (22%) also possessed the llsX gene. The majority of collected isolates (65%) belonged to molecular serogroup 1/2a-3a, followed by 4ab-4b-4d-4e (33%), and only one to serogroup 1/2b-3b-7 (2%). Isolates yielded 18 different restriction profiles, revealing a large cluster of contamination linked to frozen corn (21 strains) and distributed in 3 pulsotypes. MLST analysis classified selected isolates into nine clonal complexes (CCs). The obtained results contribute to characterizing the diversity of L. monocytogenes isolated from various vegetable products in Poland and their impact on food safety and public health. • 0.56% vegetable products in Poland were positive for L. monocytogenes. • The most prevalent was serogroup 1/2a-3a (65%). • All isolates were susceptible to tested antimicrobials. • The inlA , inlC and lmo2672 genes were observed in all isolates. • MLST analysis classified selected isolates into nine clonal complexes. [ABSTRACT FROM AUTHOR]

Highlights from the article: B BACKGROUND b : Pharmacists have been underutilized in the healthcare field, despite having a vast knowledge of drug information. B METHODS b : Adult patients with diabetes are contacted by the pharmacist or pharmacy student 3-10 days prior to the PCP visit for a comprehensive medication review focusing on diabetes.

Respiratory depression remains an important clinical problem that limits the use of opiate analgesia. Activation of AMPA glutamate receptors has been shown to reverse fentanyl-induced respiratory changes. Here, we explored whether tianeptine, a drug known for its ability to phosphorylate AMPA receptors, can be used to prevent opiate-induced respiratory depression. A model of respiratory depression in conscious rats was produced by administration of morphine (10 mg/kg, i.p.). Rats were pre-treated with test compounds or control solutions 5 min prior to administration of morphine. Respiratory activity was measured using whole-body plethysmography. In conscious animals, tianeptine (2 and 10 mg/kg, ip) and DP-201 (2-(4-((3-chloro-6-methyl-5,5-dioxido-6,11-dihydrodibenzo[c,f][1,2] thiazepin-11-yl)amino)butoxy)acetic acid; tianeptine analogue; 2 mg/kg, ip) triggered significant (~30%) increases in baseline respiratory activity and prevented morphine-induced respiratory depression. These effects were similar to those produced by an ampakine CX-546 (15 mg/kg, ip). The antinociceptive effect of morphine (hot plate test) was unaffected by tianeptine pre-treatment. In conclusion, the results of the experiments conducted in conscious rats demonstrate that systemic administration of tianeptine increases respiratory output and prevents morphine-induced respiratory depression without interfering with the antinociceptive effect of opiates. [ABSTRACT FROM AUTHOR]

In recent years, several studies have described the presence of Clostridium difficile in healthy and diarrhoeic farm and domestic animals. In pigs and cattle, the isolation of some PCR-ribotypes associated with human infection, especially PCR-ribotypes 014 and 078, has led us to hypothesize about the zoonotic transmission of C. difficile infections. If these animals are reservoirs of C. difficile , farmers in close contact with their animals are particularly at risk of acquiring and spreading the bacterium. This study investigates the presence of C. difficile in closely associated populations, beef cattle and farmers, as well as in the animal feed, manure and dust in five different farms in Belgium. C. difficile was isolated from calves and cattle with a prevalence varying between 5.5% and 11.3%. Furthermore, all of the isolates were toxigenic. An important age and breed effect was observed in the colonization of C. difficile. For age, there was a higher probability of colonization in calves of less than 6 months in age than in cattle over 11 months of age. For the type of breed a higher prevalence of the bacterium was detected in the Limousin breed than in the Belgian Bleu breed. By contrast, none of the human and animal feed samples tested positive for C. difficile . The results obtained indicate a persistent animal reservoir of C. difficile , but an indirect dissemination to humans, probably via the environment. [ABSTRACT FROM AUTHOR]

In order to control food losses and wastage, monitoring the microbial diversity of food products, during processing and storage is important, as studies have highlighted the metabolic activities of some microorganisms which can lead to spoilage. Knowledge of this diversity can be greatly improved by using a metagenetic approach based on high throughput 16S rRNA gene sequencing, which enables a much higher resolution than culture-based methods. Moreover, the Jameson effect, a phenomenon described by Jameson in 1962, is often used to classify bacterial strains within an ecosystem. According to this, we have studied the bacterial microbiota of Belgian white pudding during storage at different temperatures using culture-dependent and independent methods. The product was inoculated with a mix of dominant strains previously isolated from this foodstuff at the end of its shelf life ( Carnobacterium maltaromaticum , Lactobacillus fuchuensis , Lactobacillus graminis , Lactobacillus oligofermentans , Lactococcus lactis , Leuconostoc mesenteroides , Raoultella terrigena and Serratia sp.). Daily during 16 days, the absolute abundance of inoculated strain was monitored by combining total count on plate agar and metagenetic analysis. The results were confirmed by qPCR analysis. The growth of each species was modelled for each temperature conditions, representative of good or bad storage practices. These data allowed the bacterial strains subdivision into three classes based on criteria of growth parameters for the studied temperature: the “dominant”, the “subdominant” and the “inhibited” bacterial species, according to their maximal concentration (Nmax, log CFU/g), growth rate (μmax, 1/h) and time to reach the stationary phase (TRSP, days). Thereby, depending on the storage conditions, these data have permitted to follow intrinsically the evolution of each strain on the bacterial ecosystem of Belgian white pudding. Interestingly, it has shown that the reliability of the Jameson effect can be discussed. For example, at 4 °C when Lactococcus lactis and Serratia sp. stopped growth at day 12, at the same time Carnobacterium maltaromaticum reached its maximal concentration and entered its stationary phase. In opposition to this, it can be noticed that in the same condition, the “sub-dominant” organisms continued their growth independently of the “dominant” species behaviour. In this case, the Jameson effect was not illustrated. This pattern is described for all storage conditions with the same strain classifications. These results highlighted the importance of combining metagenetic analysis and classical methods, with modelling, to offer a new tool for studying the evolution of microorganisms present in perishable food within different environmental conditions. [ABSTRACT FROM AUTHOR]

Meat-based meals are consumed as a source of animal proteins and constitute one of the leading vehicles for food borne infections in humans. The main objective of this study was to determine the consumption pattern and the bacteriological quality of meat-based meals consumed outside households in Kigali. A survey on meat consumption patterns was carried out in 400 households by using a questionnaire, whereas different meat-based meals were sampled from 150 snack bars and restaurants. Enumeration of hygiene indicator bacteria (total mesophilic bacteria and Escherichia coli ) and the qualitative detection of Salmonella were carried out by using conventional culture methods. The results indicated that goat was the type of meat that was consumed the most outside the home in Kigali and the meat intake varied significantly (p ≤ 0.05) with the social category of the household. The average levels of total aerobic bacteria and E. coli in meat-based meals were found to be 4.7 and 1.4 log cfu/g, respectively, whereas Salmonella was detected in 11.7% of all meat-based meals. Eight factors mostly linked to the cooking treatments and hygienic handling practices for cooked meals were found to be significantly (p ≤ 0.05) associated with the risk of Salmonella occurrence in meat-based meals consumed outside the home in Kigali. The findings from this study strongly suggest the need for proper cooking and/or improvements in hygiene in the establishments selling ready-to-eat meat-based meals in Kigali, particularly those located in rural localities. [ABSTRACT FROM AUTHOR]

Meat is consumed worldwide as a source of animal proteins, but it is recognized as one of the most important vehicles for food borne infections in humans. This study was conducted to determine the daily intake; the levels of hygiene indicator bacteria, namely the total mesophilic bacteria (TMC) and Escherichia coli counts (ECC); and the prevalence of Salmonella in meat consumed within the households of Kigali (Rwanda). The survey on meat consumption was carried out in 400 households by using a questionnaire, whereas the bacteriological analyses of meat samples were performed by using conventional culture methods. The results from the survey indicated that beef was the type of meat mostly consumed in Kigali city households, and the daily meat intake significantly varied with the social category of the household. No significant difference was observed between daily meat intakes in different age classes of household members. In the samples where microorganisms were detected, the average levels of TMCs and ECCs in raw meat were found to be 5.4 and 1.6 log cfu/g, respectively, whereas in cooked meat they were significantly reduced to 3.1 and 1.1 log cfu/g, respectively. The prevalence of Salmonella was reduced from 21.4% in raw meat to 3.4% in ready-to-eat cooked meat. Salmonella was not detected in cooked meat consumed in high-income households. The results from this study highlight the need for hygiene improvements in meat shops as well as in the households of Kigali, particularly those with low and medium incomes. [ABSTRACT FROM AUTHOR]

The aim of the study was to characterize strains of Listeria monocytogenes isolated from ready to eat (RTE) products collected as part of official food control and monitoring in Poland. A total of 105 L. monocytogenes isolates from RTE products: 54- cakes and 51 – delicatessen products were examined. The presence L. monocytogenes in cakes and delicatessen products was 0.4% and 0.7% respectively suggesting the level of contamination of RTE products with L. monocytogenes is very low. [ABSTRACT FROM AUTHOR]

Herve cheese is a Belgian soft cheese with a washed rind, and is made from raw or pasteurized milk. The specific microbiota of this cheese has never previously been fully explored and the use of raw or pasteurized milk in addition to starters is assumed to affect the microbiota of the rind and the heart. The aim of the study was to analyze the bacterial microbiota of Herve cheese using classical microbiology and a metagenomic approach based on 16S ribosomal DNA pyrosequencing. Using classical microbiology, the total counts of bacteria were comparable for the 11 samples of tested raw and pasteurized milk cheeses, reaching almost 8 log cfu/g. Using the metagenomic approach, 207 different phylotypes were identified. The rind of both the raw and pasteurized milk cheeses was found to be highly diversified. However, 96.3 and 97.9% of the total microbiota of the raw milk and pasteurized cheese rind, respectively, were composed of species present in both types of cheese, such as Corynebacterium casei, Psychrobacter spp., Lactococcus lactis ssp. cremoris, Staphylococcus equorum, Vagococcus salmoninarum, and other species present at levels below 5%. Brevibacterium linens were present at low levels (0.5 and 1.6%, respectively) on the rind of both the raw and the pasteurized milk cheeses, even though this bacterium had been inoculated during the manufacturing process. Interestingly, Psychroflexus casei, also described as giving a red smear to Raclettetype cheese, was identified in small proportions in the composition of the rind of both the raw and pasteurized milk cheeses (0.17 and 0.5%, respectively). In the heart of the cheeses, the common species of bacteria reached more than 99%. The main species identified were Lactococcus lactis ssp. cremoris, Psychrobacter spp., and Staphylococcus equorum ssp. equorum. Interestingly, 93 phylotypes were present only in the raw milk cheeses and 29 only in the pasteurized milk cheeses, showing the high diversity of the microbiota. Corynebacterium casei and Enterococcus faecalis were more prevalent in the raw milk cheeses, whereas Psychrobacter celer was present in the pasteurized milk cheeses. However, this specific microbiota represented a low proportion of the cheese microbiota. This study demonstrated that Herve cheese microbiota is rich and that pasteurized milk cheeses are microbiologically very close to raw milk cheeses, probably due to the similar manufacturing process. The characterization of the microbiota of this particular protected designation of origin cheese was useful in enabling us to gain a better knowledge of the bacteria responsible for the character of this cheese. [ABSTRACT FROM AUTHOR]

Abstract: Combat operations in contemporary theaters of war, as well as combat training, are carried out in all parts of the world, typically in a harsh environment. Specific environmental conditions, such as heat, cold, high-altitudes, desert climates, as well as chemical and biological pollution of both the atmosphere and soil, together with over-exertion, food restrictions, sleep deprivation, and psychological stress can all result in changes in the immune system and the occurrence of associated diseases. Respiratory diseases are one of the most common health problems among military personnel participating in combat training or deployed to operations in areas characterized by difficult climatic and sanitary conditions. They are, therefore, one of the main reasons for military personnel requiring ambulant and hospital treatment. The aim of the study was to discuss the influence of environmental factors and the conditions in which active duty is performed on changes in the immune system and the occurrence of respiratory tract diseases in a military environment. [Copyright &y& Elsevier]

Abstract: Amphiphilic diblock copolypeptide hydrogels (DCHs) are synthetic materials whose properties can be varied readily and predictably by altering copolymer chain length or composition and which are of potential interest for biomaterial applications. We tested the biocompatibility in the central nervous system (CNS) of DCH composed of lysine, homoarginine or glutamate in combination with leucine. A range of DCH formulations with rheological properties similar to brain tissue were injected into mouse forebrain and examined after 1–8 weeks using light microscopy, immunohistochemistry and electron microscopy. DCH deposits elicited no more gliosis, inflammation, or toxicity to neurons, myelin or axons than did injections of physiological saline. The size, rigidity, and density of DCH deposits could be varied subtly by altering DCH composition and concentration. For any given DCH formulation, increased concentration correlated with increased gel strength in vitro and increased deposit size in vivo. DCHs of lysine and leucine (K m L n ) were selected for detailed analyses because these formed deposits with desirable physical properties and since lysine is routinely used as a substrate for neural cell cultures. Deposits of unmodified K180L20 exhibited time-dependent in-growth of blood vessels and of certain glial cells, and limited in-growth of nerve fibers. These findings show that DCHs are injectable, re-assemble in vivo to form 3-dimensional deposits, exhibit little or no detectable toxicity in the CNS, integrate well with brain tissue and represent a new class of synthetic biomaterials with potential for applications as depots or scaffolds in the CNS. [Copyright &y& Elsevier]

In Cambodia, freshwater aquaculture is the most important source of food production. Fresh fish meat is considered a highly perishable food that requires the use of different manipulations and preservation techniques to inhibit the proliferation of undesirable bacteria. These bacteria are naturally present in the raw product or could be acquired during manipulation by cross-contamination. Many studies worldwide have investigated the epidemiology of Clostridioides difficile (C. difficile) in food, but to date, there are no publications about the bacterium in ready-to-eat fish or descriptions in Cambodia. The objective of this study was to assess the presence of C. difficile in one of the main food supplies of this country, smoked freshwater fish, originating from different provinces. A total of 25 samples were collected directly from local markets, yielding 4 C. difficile isolates and an overall recovery rate of 16%. Most of the isolates were toxigenic and classified as rare PCR profiles, and they were resistant to clindamycin. These findings indicate contamination during handling and/or contamination of the raw fish, followed by insufficient heat treatment to kill the spores. The presence of C. difficile in smoked and dried fish implies a potential risk of human exposure, contamination and infection. • Smoked freshwater fish is the main 1 food supply in Cambodia. • Toxigenic C. difficile is present in ready to eat smoked fish. • Isolates presented resistances to clindamycin and moxifloxacin. • First isolation of C. difficile from smoked and dried freshwater fish. • First isolation of C. difficile in Cambodia. [ABSTRACT FROM AUTHOR]

Listeria monocytogenes (L. monocytogenes) is a well-known pathogenic bacterium that causes the disease listeriosis in both humans and animals. Human transmission mainly occurs via ingestion of contaminated foods and specifically affects pregnant women, newborns, elderly individuals and immunosuppressed individuals. Several outbreaks have been historically associated with the consumption of fresh raw milk and unpasteurized cheese, highlighting the role of good farm hygiene measures to reduce the probability of milk contamination. L monocytogenes is ubiquitous in the environment, and therefore, this bacterium is commonly found in silage, haylage, grazing pastures, crop fields, farmyards and even water. Faeces of wild animals, including gulls and rooks, have also been described as important vectors of the pathogen for farm animal contamination, as well as animal bedding, soils or feed bunk tanks, especially when animals are housed during indoor months. Milking lines, including filters, collectors, bulk tanks and other utensils in the room, have been described as important sites of bacterial detection. The ability of L. monocytogenes to produce biofilms and to survive in humid environments makes elimination difficult and increases its persistence in equipment and on floors, leading to high risk of milk contamination at harvest in farms. This review explores in depth the different sources of L. monocytogenes contamination described in production farms, with a special focus on ruminants, identifying the transmission vectors and analysing the applicable control measures at each stage. • Listeriosis is an important infection for humans and animals. • L.monocytogenes is ubiquitous in the farm environment. • Feedstuffs, water, surfaces and manure are the main bacterial vectors. • Control measures include biofilm cell detachment from surfaces. • Control measures must be established in the farm and its surroundings at each stage of production. [ABSTRACT FROM AUTHOR]