To culture or not to culture: careful assessment of metabarcoding data is necessary when evaluating the microbiota of a modified-atmosphere-packaged vegetarian meat alternative throughout its shelf-life period.

BACKGROUND: As the increased consumption of ready-to-eat meat alternatives is a fairly recent trend, little is known about the composition and dynamics of the microbiota present on such products. Such information is nonetheless valuable in view of spoilage and food safety prevention. Even though refrigeration and modified-atmosphere-packaging (MAP) can extend the shelf-life period, microbial spoilage can still occur in these products. In the present study, the microbiota of a vegetarian alternative to poultry-based charcuterie was investigated during storage, contrasting the use of a culture-dependent method to a culture-independent metagenetic method. RESULTS: The former revealed that lactic acid bacteria (LAB) were the most abundant microbial group, specifically at the end of the shelf-life period, whereby Latilactobacillus sakei was the most abundant species. Metabarcoding analysis, in contrast, revealed that DNA of Xanthomonas was most prominently present, which likely was an artifact due to the presence of xanthan gum as an ingredient, followed by Streptococcus and Weissella. CONCLUSIONS: Taken together, these results indicated that Lb. sakei was likely the most prominent specific spoilage organisms (SSO) and, additionally, that the use of metagenetic analysis needs to be interpreted with care in this specific type of product. In order to improve the performance of metagenetics in food samples with a high DNA matrix but a low bacterial DNA load, selective depletion techniques for matrix DNA could be explored.

[D-CBP study: Evaluation of lung cancer management times]. / Étude D-CBP : Évaluation des délais de prise en charge du cancer broncho- pulmonaire au sein du service de Pneumologie et d'Oncologie Thoracique du CHU de Caen.

INTRODUCTION: Lung cancer is the leading cause of cancer-related death. Delays may have an impact on patient survival. The objective of this study was to evaluate the diagnostic and therapeutic management times for patients admitted for lung cancer treatment in the Respiratory Department of CHU de Caen Normandie. MATERIALS AND METHODS: This is a retrospective, single-center and observational study, conducted on all patients treated for lung cancer from June 2017 to January 2018 in our department of pneumology in the Caen Normandie CHU. The main median times were investigated were: Global Time (abnormal imaging-treatment), Diagnosis time (abnormal imaging-diagnosis) and Treatment Time (diagnosis-treatment). RESULTS: One hundred and twenty-seven (127) patients were included. Median global time was 55.5 days [31,25; 393], median diagnosis time was 22 days [13; 49], and median treatment time was 24.5 days [12,25; 45]. DISCUSSION: Our treatment times are consistent with those previously published. Areas for improvement are being developed in accordance with the 2014-2019 cancer plan, in particularly the creation in our institution of a specific care pathway for patients with lung cancer.

Interaction of CP levels in maternal and nursery diets, and its effect on performance, protein digestibility, and serum urea levels in piglets.

Reduced protein levels in nursery diets have been associated with a lower risk of postweaning diarrhea, but the interaction with CP levels in maternal diet on the performance of the offspring remains unclear. The objective of this study was to determine the effect of protein content in sow gestation and piglet nursery diets on the performance of the piglets until slaughter. This was studied in a 2 × 2 factorial trial (35 sows, 209 piglets), with higher or lower (H or L) dietary CP in sow diets (168 vs 122 g CP/kg) during late gestation. A standard lactation feed was provided for all sows (160 g CP/kg). For both sow treatments, half of the litters received a higher or lower CP in the piglet nursery diet (210 vs 166 g CP/kg). This resulted in four possible treatment combinations: HH, HL, LH and LL, with sow treatment as first and piglet treatment as second letter. For each phase, all diets were iso-energetic and had a similar level of essential amino acids. Ps*p is the p-value for the interaction effect between sow and piglet treatment. In the nursery phase (3.5-9 weeks of age), a tendency toward interaction between piglet and sow treatments with feed efficiency (Ps*p = 0.08) was observed with HH having the highest gain:feed ratio (G:F) (0.74 ± 0.01), LH the lowest (0.70 ± 0.01) and the other two groups intermediate. In the growing-finishing phase, an interaction was observed between the piglet and sow diets with decreased G:F for LH (Ps*p = 0.04) and a tendency toward interaction with increased daily feed intake for LH (Ps*p = 0.07). The sow diet showed a tendency toward a long-lasting effect on the dressing percentage and meat thickness of the offspring, which was higher for the progeny of H sows (Ps < 0.01 and Ps = 0.02, respectively). At 23 weeks, serum urea concentrations tended to be lower for the HH and LL groups (Ps*p = 0.07). Fecal consistency scores were higher at day 10-day 14 after weaning for piglets from L sows (Ps = 0.03 and Ps < 0.01, respectively). At day 7 after weaning, fecal consistency score was higher for piglets fed the higher protein diet (Pp < 0.01). At 8 weeks of age, the apparent total tract digestibility of CP (ATTDCP) interacted between piglet and sow diet (Ps*p = 0.02), with HH showing the highest digestibility values. In conclusion, the protein levels in sow late-gestation and piglet nursery diets interacted with feed efficiency, ATTDCP and serum urea concentrations in the nursery phase.

The effect of partial depopulation on Campylobacter introduction in broiler houses.

Poultry is seen as the main reservoir for Campylobacter. Control of this zoonotic pathogen in primary production could potentially reduce the colonization in broiler flocks and consequently reduce the number of human infections. In the present study, 20 broiler flocks from 10 farms, were sampled immediately before and 5 to 7 d after partial depopulation (thinning) for the presence of Campylobacter using cecal droppings and overshoes. At the time of thinning, the catching crew, transportation vehicles, forklift, and transport containers were sampled for the presence of Campylobacter. Samples were cultivated; presumed positive isolates were confirmed by PCR. The isolates were molecularly typed by flaA restriction analysis and pulsed field gel electrophoresis. Results show that all flocks were thinned using Campylobacter-contaminated equipment and materials. One-third of the broiler flocks became colonized after thinning. In 67% of the colonization cases, identical strains were found matching those of container systems, transport trucks, and/or forklifts. This identifies thinning as an important risk factor for Campylobacter introduction into broiler houses. Setup and compliance with biosecurity practices during thinning is essential to prevent Campylobacter colonization of broiler flocks.

Pathogenic potential of Escherichia coli O157 and O26 isolated from young Belgian dairy calves by recto-anal mucosal swab culturing.

AIMS: The purpose of this study was to investigate the occurrence of Escherichia coli O157 and O26 on Belgian dairy cattle farms, the presence of virulence genes in the confirmed isolates and the association of E. coli O26 presence with calf diarrhoea. METHODS AND RESULTS: In total, 233 recto-anal mucosal swabs (RAMS) were obtained from healthy and diarrheic dairy calves on three farms, each alternately visited three consecutive times. RAMS were analysed for presence of E. coli O157 and O26, and stx1, stx2 and eae virulence genes. Overall, 19% of RAMS tested positive for E. coli O157, while 31% tested positive for E. coli O26. The majority of isolates possessed both stx and eae, denoting a high pathogenic potential to humans. While both serogroups persisted at farm level, persistence within the same animal over time appeared to be relatively rare. Interestingly, E. coli O26 was already abundantly present at a younger age compared to E. coli O157. Calf diarrhoea could not be associated with presence of E. coli O26. CONCLUSIONS: Young dairy calves are important on-farm reservoirs of potentially pathogenic E. coli O157 and O26. A role of E. coli O26 in calf diarrhoea could not be confirmed. SIGNIFICANCE AND IMPACT OF THE STUDY: O157 and O26 are responsible for the majority of human STEC infections. Gaining more epidemiological information regarding their occurrence and persistence on cattle farms will contribute to a better understanding of STEC ecology and risk of human transmission.

Evaluation of group vaccination of sows and gilts against Salmonella Typhimurium with an attenuated vaccine in subclinically infected pig herds.

Subclinical Salmonella Typhimurium infections occur frequently in pigs and constitute a major risk for human salmonellosis. With the currently available control measures, Salmonella Typhimurium infections in pigs remain difficult to control. Vaccination has been proposed to be an effective tool to control infections at farm level. In the current study, the effect of group vaccination of sows and gilts against Salmonella Typhimurium is evaluated on Salmonella prevalence in fecal and overshoe samples and ileocecal lymph nodes, and on serology in the sows and their offspring in three subclinically infected pig farms. In each farm, all sows and gilts were vaccinated twice, three weeks apart, with an attenuated histidine-adenine auxotrophic vaccine (Salmoporc®, IDT Biologika). From three months after the group vaccination onwards, all sows were given a booster dose three weeks before every farrowing. The farms were monitored bacteriologically and serologically from 12 months before until 15 months after the group vaccination. After group vaccination, no significant effect was detected in the prevalence of Salmonella Typhimurium in the fecal and overshoe samples collected in the sows (before: 2 %, after: 0 %) and their offspring at 18 weeks (before: 17 %, after: 11 %) and at 26 weeks of age (before: 15 %, after: 7 %), and when combining the results of the offspring at 18 and 26 weeks of age (before: 16 %, after: 9 %). Also, no significant effect was detected in the prevalence of Salmonella Typhimurium positive lymph nodes of sows (before and after: 0 %) and their offspring (before: 4 %, after: 7 %). Regarding serology, the mean S/P-ratios of the sows were significantly higher after the group vaccination, compared to before group vaccination (before: 1.50, after: 2.32, p < 0.001). The mean S/P-ratios of the offspring at slaughter age were significantly lower after the group vaccination, compared to before group vaccination (before: 1.71, after: 1.04, p = 0.001). In conclusion, group vaccination of sows and gilts resulted in a more beneficial serological status of the offspring, but did not significantly decrease Salmonella Typhimurium excretion and lymph node contamination.

Monitoring of Hygiene in Institutional Kitchens in Belgium.

ABSTRACT: Microbiological contamination of food during preparation and storage is a risk factor in institutional kitchens. In this Belgian study, hygiene practices in 40 institutional kitchens from four public sectors (10 hospitals, 10 schools, 10 retirement homes, and 10 child care centers) were evaluated to determine whether differences in these practices exist between these sectors. Contamination levels were also analyzed at several critical contact points. A data collection instrument and microbiological analysis of hand contact surfaces, food contact surfaces, and kitchen utensils were used. Hand washing resulted in only a slight reduction in total aerobic bacteria counts (TACs), and all microorganisms evaluated except E. coli were still present at countable levels. Enterobacteriaceae were found on one-third of the cleaned cutting boards. Cleaned work surfaces had the highest average TAC of all cleaned surfaces. Only slight improvements in TACs and Enterobacteriaceae and B. cereus counts were observed between used and cleaned work surfaces. The results from the data collection instrument revealed that child care centers had the lowest hygiene scores, whereas the other three sectors were fairly similar, with hospitals scoring highest. The low hygiene score for the child care centers was verified by comparing the results for cleaned surfaces among the sectors. The average TAC on surfaces was highest for child care centers and lowest for hospitals. Child care centers also had the second highest total mean counts and the highest number of total surface samples positive for Enterobacteriaceae. The highest number of surface samples positive for Staphylococcus aureus was also found in child care centers. This study highlights some areas of concern for hygiene improvement in institutional kitchens, differences between public sectors, and similarities in conclusions about hygiene based on the scores from the survey instrument and the results of the microbiological analyses.

Bacteriological evaluation of vaccination against Salmonella Typhimurium with an attenuated vaccine in subclinically infected pig herds.

Subclinical infections with Salmonella Typhimurium occur frequently in pigs. They constitute a risk for human salmonellosis and are difficult to control with currently available control measures. Vaccination against Salmonella Typhimurium in pigs can be an effective tool to control Salmonella infections at farm level. In the present study, the efficacy of an attenuated Salmonella Typhimurium vaccine (Salmoporc®, IDT Biologika) to control Salmonella infections in pigs was evaluated in three subclinically infected pig herds. The effect on Salmonella excretion and the number of pigs positive for Salmonella Typhimurium field and vaccine strains in ileocecal lymph nodes at slaughter were evaluated using five different vaccination strategies: 1. vaccination of sows, 2. vaccination of sows and piglets, 3. vaccination of sows and fattening pigs, 4. vaccination of piglets, 5. vaccination of fattening pigs, which were all compared to a non-vaccinated control group (experimental group 6). Each vaccination strategy was implemented in each farm, during two consecutive production cycles of the same sows. The prevalence of Salmonella Typhimurium field strain excretion was low; in total, 4% of the fecal and overshoe samples collected in the non-vaccinated control group were Salmonella Typhimurium field strain positive. The excretion of Salmonella Typhimurium field strain did not significantly differ between farms, production cycles and experimental groups. Applying vaccination in either sows and piglets, sows and fattening pigs, or in piglets only, resulted in a significantly reduced number of Salmonella Typhimurium field strain positive lymph nodes of slaughter pigs in the second production cycle, but not in the first production cycle. Vaccination of sows and piglets resulted in the most consistent reduction of Salmonella Typhimurium field strain positive lymph nodes at slaughter. The vaccine strain was detected in the lymph nodes of 13 pigs at slaughter, indicating the possible persistence of the vaccine strain until slaughter. Because of limitations in the study design, and the variability between farms and production cycles, the results of the current observational study should be extrapolated with care. Nevertheless, the results provide evidence that applying vaccination against Salmonella Typhimurium in sows and piglets (preferred), sows and fattening pigs, and piglets only can support the control of Salmonella Typhimurium infections by decreasing the prevalence of Salmonella Typhimurium field strain positive lymph nodes at slaughter.

Effect of a pre-milking teat foam and a liner disinfectant on the presence of mesophilic and (proteolytic) psychrotrophic bacteria prior to milking.

Contamination of raw milk by psychrotrophs can lead to the production of heat-resistant proteases and subsequent spoilage of UHT milk. Therefore, this research communication evaluated the effect of a pre-milking teat disinfectant (active components: L-(+)-lactic acid and salicylic acid) and a liner disinfectant (active components: peracetic acid and hydrogen peroxide) on the number of mesophilic and (proteolytic) psychrotrophic bacteria prior to milking. The teat orifices of 10 cows were sampled using a swabbing procedure before and after treatment with a pre-milking teat disinfectant on six subsequent days. On the teat orifices, there was a small but statistically significant decrease in the psychrotrophic bacterial counts between pre and post dipping. No differences were observed for the mesophilic bacterial counts and proteolytic active counts. Liners were also sampled using swabs pre and post disinfection. No statistically significant decrease in the bacterial counts was observed post liner disinfection, although there was a numerical decrease. Sixty-two percent of the proteolytic psychrotrophs were pseudomonads: 16.5% of which were P. fragi, 14.3% P. lundensis, 10.0% P. fluorescens and 2.9% P. putida. Trinitrobenzenesulfonic acid (TNBS) analysis revealed a wide variety in proteolytic activity (from 0 to 55 µmol glycine/ml milk) and the presence of high producers. It can be concluded that there was only a minor effect of teat and liner disinfection on the psychrotrophic bacterial counts indicating that the measures presented did not result in a reduction of the targeted bacteria on teat orifices and liners.

Effects of attenuated vaccine protocols against Salmonella Typhimurium on Salmonella serology in subclinically infected pig herds.

Vaccination of pigs against Salmonella Typhimurium (S. Typhimurium) can be effective for the control of Salmonella infections at the farm level and reduce the risk of Salmonella contamination in the food chain. However, vaccination may interfere with herd serological status in serology-based Salmonella monitoring programs. The present study investigated the effects of an attenuated S. Typhimurium vaccine (Salmoporc, IDT Biologika) on Salmonella serology in sows, neonatal piglets and slaughter pigs from three subclinically infected herds. Within each herd, five different vaccination protocols were tested as follows: group 1, vaccination of sows; group 2, vaccination of sows and piglets; group 3, vaccination of sows and fattening pigs; group 4, vaccination of piglets; and group 5 vaccination of fattening pigs. Each group was compared to a non-vaccinated control group (group 6). Sera were analyzed by ELISA (HerdChek Swine Salmonella, IDEXX Laboratories) and sample-to-positive (S/P) ratios were calculated. At day 3 after farrowing, but not before vaccination, S/P ratios in vaccinated sows (mean: 2.21) were significantly higher than S/P ratios in non-vaccinated sows (mean: 0.87, P<0.001). S/P ratios in 3-day old piglets from vaccinated sows (mean: 2.46) were significantly higher than S/P ratios in similar piglets from non-vaccinated sows (mean: 0.73, P<0.001). At slaughter, S/P ratios in pigs from groups 2, 3, 4 and 5 were significantly higher than those in the non-vaccinated control group (P<0.001). Therefore, vaccination of piglets and fattening pigs could have implications for current serology-based Salmonella monitoring programs in slaughter pigs.

Selection and transfer of an IncI1-tet(A) plasmid of Escherichia coli in an ex vivo model of the porcine caecum at doxycycline concentrations caused by crosscontaminated feed.

AIMS: The aim of this study was to investigate the effect of subtherapeutic intestinal doxycycline (DOX) concentrations (4 and 1 mg l-1 ), caused by cross-contamination of feed, on the enrichment of a DOX-resistant commensal Escherichia coli and its resistance plasmid in an ex vivo model of the porcine caecum. METHODS AND RESULTS: A DOX-resistant, tet(A)-carrying, porcine commensal E. coli strain (EC 682) was cultivated for 6 days in the porcine caecum model under different conditions (0, 1 and 4 mg l-1 DOX). EC 682, other coliforms and anaerobic bacteria were enumerated daily. A selection of isolated DOX-resistant coliforms (n = 454) was characterized by rep-PCR clustering, PCR assays (Inc1 and tet(A)) and micro broth dilution susceptibility tests (Sensititre). Both 1 and 4 mg l-1 DOX-enriched medium had a significantly higher selective effect on EC 682 and other resistant coliforms than medium without DOX. Transconjugants of EC 682 were isolated more frequently in the presence of 1 and 4 mg l-1 DOX compared to medium without DOX. CONCLUSIONS: Subtherapeutic intestinal DOX concentrations have the potential to select for DOX-resistant E. coli, and promote the selection of transconjugants in a porcine caecum model. SIGNIFICANCE AND IMPACT OF THE STUDY: Cross-contamination of feed with antimicrobials such as DOX likely promotes the spread of antimicrobial resistance. Therefore, it is important to develop or fine-tune guidelines for the safe use of antimicrobials in animal feed and its storage.

Reduction of Mycobacterium avium ssp. paratuberculosis in colostrum: Development and validation of 2 methods, one based on curdling and one based on centrifugation.

The aim of this study was to develop and validate 2 protocols (for use on-farm and at a central location) for the reduction of Mycobacterium avium ssp. paratuberculosis (MAP) in colostrum while preserving beneficial immunoglobulins (IgG). The on-farm protocol was based on curdling of the colostrum, where the IgG remain in the whey and the MAP bacteria are trapped in the curd. First, the colostrum was diluted with water (2 volumes colostrum to 1 volume water) and 2% rennet was added. After incubation (1 h at 32°C), the curd was cut and incubated again, after which whey and curd were separated using a cheesecloth. The curd was removed and milk powder was added to the whey. Approximately 1 log reduction in MAP counts was achieved. A reduction in total proteins and IgG was observed due to initial dilution of the colostrum. After curd formation, more than 95% of the immunoglobulins remained in the whey fraction. The semi-industrial protocol was based on centrifugation, which causes MAP to precipitate, while the IgG remain in the supernatant. This protocol was first developed in the laboratory. The colostrum was diluted with skimmed colostrum (2 volumes colostrum to 1 volume skimmed colostrum), then skimmed and centrifuged (at 15,600 × g for 30 min at room temperature). We observed on average 1.5 log reduction in the MAP counts and a limited reduction in proteins and IgG in the supernatant. To obtain a semi-industrial protocol, dairy pilot appliances were evaluated and the following changes were applied to the protocol: after 2:1 dilution as above, the colostrum was skimmed and subsequently clarified, after which the cream was heat treated and added to the supernatant. To investigate the effect of the colostrum treatment on the nutritional value and palatability of the colostrum and the IgG transfer, an animal experiment was conducted with 24 calves. Six received the dam's colostrum, 6 were given untreated purchased colostrum (control), and 2 groups of 6 calves received colostrum treated according to both of the above-mentioned methods. No significant differences were found between the test groups and the dam's colostrum group in terms of animal health, IgG uptake in the blood serum, milk, or forage uptake. Two protocols to reduce MAP in colostrum (for use on-farm or at a central location) were developed. Both methods preserve the vital IgG.

A 10-day vacancy period after cleaning and disinfection has no effect on the bacterial load in pig nursery units.

BACKGROUND: Biosecurity measures such as cleaning, disinfection and a vacancy period between production cycles on pig farms are essential to prevent disease outbreaks. No studies have tested the effect of a longer vacancy period on bacterial load in nursery units. METHODS: The present study evaluated the effect of a 10-day vacancy period in pig nursery units on total aerobic flora, Enterococcus spp., Escherichia coli, faecal coliforms and methicillin resistant Staphylococcus aureus (MRSA). Three vacancy periods of 10 days were monitored, each time applied in 3 units. The microbiological load was measured before disinfection and at 1, 4, 7 and 10 days after disinfection. RESULTS: No significant decrease or increase in E. coli, faecal coliforms, MRSA and Enterococcus spp. was noticed. Total aerobic flora counts were the lowest on day 4 after disinfection (i.e. 4.07 log CFU/625 cm2) (P < 0.05), but the difference with other sampling moments was limited (i.e. 0.6 log CFU/625 cm2) and therefore negligible. Furthermore, this observation on day 4 was not confirmed for the other microbiological parameters. After disinfection, drinking nipples were still mostly contaminated with total aerobic flora (i.e. 5.32 log CFU/625 cm2) and Enterococcus spp. (i.e. 95 % of the samples were positive) (P < 0.01); the feeding troughs were the cleanest location (total aerobic flora: 3.53 log CFU/625 cm2 and Enterococcus spp.: 50 % positive samples) (P < 0.01). CONCLUSIONS: This study indicates that prolonging the vacancy period in nursery units to 10 days after disinfection with no extra biosecurity measures has no impact on the environmental load of total aerobic flora, E. coli, faecal coliforms, MRSA and Enterococcus spp..

Comparison of competitive exclusion with classical cleaning and disinfection on bacterial load in pig nursery units.

BACKGROUND: Colonisation of the environment of nursery units by pathogenic micro-organisms is an important factor in the persistence and spread of endemic diseases in pigs and zoonotic pathogens. These pathogens are generally controlled by the use of antibiotics and disinfectants. Since an increasing resistance against these measures has been reported in recent years, methods such as competitive exclusion (CE) are promoted as promising alternatives. RESULTS: This study showed that the infection pressure in CE units after microbial cleaning was not reduced to the same degree as in control units. Despite sufficient administration of probiotic-type spores, the analysed bacteria did not decrease in number after 3 production rounds in CE units, indicating no competitive exclusion. In addition, no differences in feed conversion were found between piglets raised in CE and control units in our study. Also, no differences in faecal consistency (indicator for enteric diseases) was noticed. CONCLUSION: These results indicate that the CE protocol is not a valuable alternative for classical C&D.

Case report of Salmonella cross-contamination in a food laboratory.

BACKGROUND: This paper describes a case of Salmonella cross-contamination in a food laboratory. In 2012, chocolate bars shipped from Belgium to the USA were prevented from entering the USA because a Salmonella Rissen strain had been isolated from one of the chocolate bars in a Belgian food laboratory. However, a retrospective study of the Salmonella isolates sent from the laboratory to the Belgian National Reference Laboratory for Salmonella revealed that 7 weeks prior, a Salmonella Rissen strain has been isolated from fish meal in the same food laboratory. The chocolate bars were not expected to be contaminated with Salmonella because the ingredients all tested negative during the production process. Furthermore, because Salmonella Rissen is only rarely isolated from food, it was hypothesized that the two Salmonella Rissen isolates belonged to the same strain and that the second isolation event in this laboratory was caused by cross-contamination. To confirm this hypothesis, both Salmonella Rissen isolates were fingerprinted using different molecular techniques. To evaluate the discriminatory power of the techniques used, 11 other Salmonella Rissen isolates from different origins were included in the comparison. Pulsed-field gel electrophoresis, repetitive element palindromic PCR and three random amplified polymorphic DNA PCR assays were used. RESULTS: Repetitive element palindromic PCR and random amplified polymorphic DNA PCR assays were insufficiently discriminatory, whereas pulsed-field gel electrophoresis using the combination of two restriction enzymes showed sufficient discrimination to confirm the hypothesis. CONCLUSIONS: Although cross-contamination in food laboratories are rarely reported, cross-contamination can always occur. Laboratories should therefore always be aware of the possibility of cross-contamination, especially when enrichment is used in the microbiological analysis. Furthermore, it is advised that results showing isolates of the same serotype isolated in a short time frame from unrelated food products should be interpreted carefully and should be confirmed with additional strain typing.

Spoilage potential of Vagococcus salmoninarum in preservative-free, MAP-stored brown shrimp and differentiation from Brochothrix thermosphacta on streptomycin thallous acetate actidione agar.

AIMS: During a previous study concerning brown shrimp (Crangon crangon), selective streptomycin thallous acetate actidione (STAA) agar was used to determine the growth of Brochothrix thermosphacta. However, the growth of Vagococcus salmoninarum on this medium was also noticed. This study explores the spoilage potential of this organism when inoculated on sterile shrimp. METHODS AND RESULTS: Isolates growing on STAA were identified using (GTG)5 clustering followed by partial 16S rRNA gene sequence analysis. Their biochemical spoilage potential was analysed for H2 S production and enzymatic activities were tested using an APIZYM test. Headspace solid phase micro-extraction (SPME) and gas chromatography-mass spectrometry (GC-MS) were used to analyse the volatile organic compounds (VOCs) produced during storage of inoculated shrimp. CONCLUSION: Fifty-five per cent of isolates taken from STAA could be identified as V. salmoninarum, while no apparent morphological difference with B. thermosphacta isolates was identified upon the prescribed incubation conditions. For isolates identified as V. salmoninarum, production of 2-heptanone, 2-nonanone, 2-undecanone was found, as was the possibility to form H2 S. SIGNIFICANCE AND IMPACT OF THE STUDY: When using the STAA medium for detecting B. thermosphacta, one should consider the possible abundant presence of V. salmoninarum as well. Based on this study, V. salmoninarum does not exhibit great spoilage potential, although it can produce H2 S and formed VOCs which are also found in other spoiled seafood products.

Effect of Organic Acids on Salmonella Shedding and Colonization in Pigs on a Farm with High Salmonella Prevalence.

This study builds on the results of a previous study in which six commercial feed products based on organic acids were evaluated with respect to Salmonella contamination of piglets in an artificially challenged seeder model. In the present study, the efficacy of three of these commercial products was assessed for Salmonella reduction in fattening pigs on one closed farm with a natural high Salmonella prevalence. In each of four fattening compartments, one of the following feed treatments was evaluated during two consecutive fattening rounds: (i) butyric acid (active ingredients at 1.3 kg/ton of feed; supplement A1), (ii) a combination of short-chain organic acids (mixture of free acids and salts) and natural extracts (2.92 kg/ton; supplement A4), (iii) a 1:1 blend of two commercial products consisting of medium-chain fatty acids, lactic acid, and oregano oil (3.71 kg/ton; supplement A5+A6), and (iv) a control feed. On the farm, the Salmonella status of the fattening pigs was evaluated by taking fecal samples twice during the fattening period. At the slaughterhouse, samples were collected from the cecal contents and the ileocecal lymph nodes. Salmonella isolates were serotyped and characterized by pulsed-field gel electrophoresis. This farm had a particularly high number of pigs shedding Salmonella with a wide variety of sero- and pulsotypes. Only the feed blend based on the medium-chain fatty acids was able to significantly reduce Salmonella prevalence both on the farm and at the slaughterhouse. With this combined supplement, the Salmonella reduction in the feces at slaughter age, in cecal contents at slaughter, and the lymph nodes was 50, 36, and 67%, respectively, compared with the control animals. This promising finding calls for further investigation including cost-efficiency of this combined feed product and its effect on the animals.

On-farm comparisons of different cleaning protocols in broiler houses.

The present study evaluated the effectiveness of 4 cleaning protocols designed to reduce the bacteriological infection pressure on broiler farms and prevent food-borne zoonoses. Additionally, difficult to clean locations and possible sources of infection were identified. Cleaning and disinfection rounds were evaluated in 12 broiler houses on 5 farms through microbiological analyses and adenosine triphosphate hygiene monitoring. Samples were taken at 3 different times: before cleaning, after cleaning, and after disinfection. At each sampling time, swabs were taken from various locations for enumeration of the total aerobic flora and Enterococcus species pluralis ( SPP:). In addition, before cleaning and after disinfection, testing for Escherichia coli and Salmonella was carried out. Finally, adenosine triphosphate swabs and agar contact plates for total aerobic flora counts were taken after cleaning and disinfection, respectively. Total aerobic flora and Enterococcus spp. counts on the swab samples showed that cleaning protocols which were preceded by an overnight soaking with water caused a higher bacterial reduction compared to protocols without a preceding soaking step. Moreover, soaking of broiler houses leads to less water consumption and reduced working time during high pressure cleaning. No differences were found between protocols using cold or warm water during cleaning. Drinking cups, drain holes, and floor cracks were identified as critical locations for cleaning and disinfection in broiler houses.

Comparison of sampling procedures and microbiological and non-microbiological parameters to evaluate cleaning and disinfection in broiler houses.

Cleaning and disinfection of the broiler stable environment is an essential part of farm hygiene management. Adequate cleaning and disinfection is essential for prevention and control of animal diseases and zoonoses. The goal of this study was to shed light on the dynamics of microbiological and non-microbiological parameters during the successive steps of cleaning and disinfection and to select the most suitable sampling methods and parameters to evaluate cleaning and disinfection in broiler houses. The effectiveness of cleaning and disinfection protocols was measured in six broiler houses on two farms through visual inspection, adenosine triphosphate hygiene monitoring and microbiological analyses. Samples were taken at three time points: 1) before cleaning, 2) after cleaning, and 3) after disinfection. Before cleaning and after disinfection, air samples were taken in addition to agar contact plates and swab samples taken from various sampling points for enumeration of total aerobic flora, Enterococcus spp., and Escherichia coli and the detection of E. coli and Salmonella. After cleaning, air samples, swab samples, and adenosine triphosphate swabs were taken and a visual score was also assigned for each sampling point. The mean total aerobic flora determined by swab samples decreased from 7.7±1.4 to 5.7±1.2 log CFU/625 cm2 after cleaning and to 4.2±1.6 log CFU/625 cm2 after disinfection. Agar contact plates were used as the standard for evaluating cleaning and disinfection, but in this study they were found to be less suitable than swabs for enumeration. In addition to measuring total aerobic flora, Enterococcus spp. seemed to be a better hygiene indicator to evaluate cleaning and disinfection protocols than E. coli. All stables were Salmonella negative, but the detection of its indicator organism E. coli provided additional information for evaluating cleaning and disinfection protocols. Adenosine triphosphate analyses gave additional information about the hygiene level of the different sampling points.