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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 Al), (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. [ABSTRACT FROM AUTHOR]

The objective of the study was to evaluate the combined effects of pasteurization intensity (no heat treatment and 10min at 70, 80 and 90°C), water activity (a w) (0.960–0.990), pH (5.5–7.0) and storage temperature (7 and 10°C) on the survival and outgrowth of psychrotolerant spores of Bacillus cereus FF119b and Bacillus pumilus FF128a. The experiments were performed in both artificial media and a validation was performed on real food products (cream, béchamel sauce and mixed vegetable soup). It was determined that in general, heat treatments of 10min at 70°C or 80°C activated the spores of both B. cereus FF119b and B. pumilus FF128a, resulting in faster outgrowth compared to native (non-heat treated) spores. A pasteurization treatment of 10min at 90°C generally resulted in the longest lag periods before outgrowth of both isolates. Some of the spores were inactivated by this heat treatment, with more inactivation being observed the lower the pH value of the heating medium. Despite this, it was also observed that under some conditions the remaining (surviving) spores were actually activated as their outgrowth took place after a shorter period of time compared to native non-heated spores. While the response of B. cereus FF119b to the pasteurization intensity in cream and béchamel sauce was similar to the trends observed in the artificial media at 10°C, in difference, outgrowth was only observed at 7°C in both products when the spores had been heated for 10min at 80°C. Moreover, no inactivation was observed in cream or béchamel sauce when the spores were heated for 10min at 90°C in these two products. This was attributed to the protective effect of fat in the cream and the ingredients in the béchamel sauce. The study provides some insight into the potential microbial (stability and safety) consequences of the current trend towards milder heat treatments which is being pursued in the food industry. [ABSTRACT FROM AUTHOR]

Salmonella represents a major challenge to the pig industry, as pork presents a risk for human salmonellosis. In this study, we have examined the effect of farm type on the prevalence of fattening pigs shedding Salmonella on 12 farms at risk for harboring Salmonella. On six open (grow-to-finish) and six closed (farrow-to-finish) farms, the prevalence of pigs shedding Salmonella was determined on two occasions approximately 2 months apart. The serovar, phage type, and antimicrobial resistance of the obtained Salmonella isolates were determined. On all farms, pigs shedding Salmonella were detected on at least one of the two sampling days. The mean within-herd prevalence was 7.8%. Closed farms were two times less likely to have pigs shedding Salmonella than open farms. On open farms, the odds of finding Salmonella shedding in pigs were 1.9 times higher when sampling was performed at slaughter age than when samples were taken halfway through the fattening period. Salmonella enterica serovar Typhimurium was the most predominant serotype, with a prevalence of 62 to 63% on both farm types. Of all the Salmonella Typhimurium isolates, 65% had the tetraresistant profile ASSuT (ampicillin, streptomycin, sulfonamide, and tetracycline) with or without additional resistance to trimethoprim-sulfonamide. Phage type DT120 seemed to be especially associated with this antimicrobial-resistant profile. The prevalence of Salmonella Typhimurium isolates showing resistance to ampicillin, streptomycin, tetracycline, sulfonamide, trimethoprim-sulfonamide, and lincomycin hydrochloride and spectinomycin sulfate tetrahydrate was significantly higher on open farms than on closed farms. [ABSTRACT FROM AUTHOR]

Abstract: The major objectives of this study were to determine the incidence, diversity and characteristics of Bacillus cereus group spp. isolated from food products marketed in Belgium. The food products investigated in this study included cooked pasta, lasagna, béchamel sauce, bolognaise sauce, fresh minced beef, fresh-cut vegetables and raw basmati rice. B. cereus group spp. were detected in 56.3% (324 of 575) of the samples giving rise to 380 strains. The highest incidence (100%) occurred in the raw basmati rice. Although only 10 (2.6%) of the 380 isolates were determined to be psychrotolerant (able to grow at ≤7°C), 25 (6.2%), 189 (49.7%) and 334 (87.9%) isolates were able to grow at mild temperature abuse conditions of 8°C, 9°C and 10°C, respectively. The large diversity of the isolates obtained (overall and between isolates obtained from the same product type) was highlighted by the results of the (GTG)5 PCR fingerprinting of 80 selected isolates. Sixty-one of these 80 isolates belonged to 15 distinct clusters (≥85% Pearson correlation) whereas the remaining 19 were each clustered separately. Further diversity was also found in the distribution of toxin genes as 16 different profiles were observed in the 80 selected isolates. Whilst none of 80 selected strains harboured the ces gene required for the production of the emetic toxin cereulide, 42 strains (52.5%) carried all seven genes required for the production of the diarrhoeal enterotoxins: haemolytic BL, non-haemolytic enterotoxin and cytotoxin K. The results of this study highlight not only the omnipresence but also the highly diverse ecology of B. cereus spp. within and across several food product types available on the retail market in Belgium. They should also provide the impetus for more studies to enable detailed risk assessment studies to be performed. [Copyright &y& Elsevier]

Summary: A case of abomasal ulceration in a 3-month-old Belgian Blue calf is described. Microscopical examination revealed the ulcers to be demarcated by a band of neutrophilic inflammation that separated underlying healthy tissue from the superficial fibrinous necrotic material in which bacteria were present. Clostridium perfringens type A was isolated from multiple ulcers and from the intestinal contents of the animal and pulsed field gel electrophoresis confirmed that the isolates comprised a genetically clonal population. [ABSTRACT FROM AUTHOR]

Eighteen Belgian broiler flocks were followed from the hatchery to the slaughterhouse by a multiple typing approach (sero-, geno-, and phage types) for the investigation of the transmission of Salmonella and its subtypes. For 12 of the 18 flocks, there was no correlation between the serotypes found preharvest and those isolated from the feces in the transport crates and on the carcasses in the slaughterhouse. Serotypes found in the crates were usually also found on the carcasses. In 5 of the 10 flocks with Salmonella-positive broilers, complex contamination patterns with the involvement of different serotypes, genotypes, or both were revealed. In two of these flocks (flocks 8 and 9), the Salmonella Enteritidis contamination of the broilers could be traced to the hatchery. In flock 9, evidence was found for the acquisition, during rearing, of a megaplasmid in the Salmonella Enteritidis strain. In the other three positive flocks (flocks 6, 7, and 10), the environment and movable material (e.g., footwear) played a determining role in the infection and shedding pattern of the broilers. For Pocks 6 and 7, reared consecutively in the same broiler house, a persistent Salmonella Hadar geno/phage type predominated in the preharvest period, while another Salmonella Hadar geno/phage type was found in the house or the environment but never in the broilers. Only for the above-mentioned five flocks were the same strains that were found preharvest also recovered from the carcasses. although these strains were not predominant on the carcasses, with the exception of one flock (flock 10). In conclusion, it can be said that most of the time, Salmonella strains that contaminate Belgian broiler carcasses do not predominate in the preharvest environment. [ABSTRACT FROM AUTHOR]

Abstract: Thermotolerant Campylobacter jejuni and C. coli are one of the major causes of bacterial foodborne enteric infection. Consuming and/or handling poultry meat is the most consistent risk factor, linked to the high prevalence of campylobacters in retail poultry meat. The aim of the present study was to ascertain the genetic diversity and/or possible specificity of thermotolerant Campylobacter isolates according to species (C. coli, C. jejuni), isolation source (retail chicken meat and human clinical samples) and geographic origin (Goriška in Slovenia and Zenica-Doboj Canton in Bosnia and Herzegovina (BIH)). With the pulsed-field gel electrophoresis (PFGE) after SmaI macrorestriction we distinguished 80 PFGE types among 118 strains and CfoI restriction fragment length polymorphism of the amplified flagellin gene (fla-RFLP) gave 12 fla-RFLP types. Beside the higher discriminatory power and strain typeability, PFGE discriminated the C. jejuni and C. coli groups of isolates. A high proportion of C. coli strains was isolated, especially from poultry samples. Identical or very similar PFGE types among the isolates from animal, food and human samples indicate the transmission of C. jejuni and C. coli from the chickens on the farm to the retail chicken meat, as well as possible cross-contamination of retail meat and transmission to humans. However, the identity of the isolates from non-related samples but with identical PFGE and fla-RFLP types should be confirmed with additional typing. Reliable tracing of the source of Campylobacter strains by molecular typing of the chicken meat isolates is therefore very difficult. The reasons include contamination of meat samples with multiple strains, possible cross-contamination and extreme heterogeneity of the isolates (mainly for C. jejuni) on one side and a limited power of the genotyping methods used to distinguish non-related strains on the other side (mainly for C. coli). [Copyright &y& Elsevier]

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. P s*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 (P s*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 (P s*p = 0.04) and a tendency toward interaction with increased daily feed intake for LH (P s*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 (P s < 0.01 and P s = 0.02, respectively). At 23 weeks, serum urea concentrations tended to be lower for the HH and LL groups (P s*p = 0.07). Fecal consistency scores were higher at day 10–day 14 after weaning for piglets from L sows (P s = 0.03 and P s < 0.01, respectively). At day 7 after weaning, fecal consistency score was higher for piglets fed the higher protein diet (P p < 0.01). At 8 weeks of age, the apparent total tract digestibility of CP (ATTD CP) interacted between piglet and sow diet (P s*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, ATTD CP and serum urea concentrations in the nursery phase. [ABSTRACT FROM AUTHOR]

The major objective of this study was to evaluate and model the combined effect of the water activity (aw) and pH of the heating menstrum on the heat resistance of spores of a psychrotolerant Bacillus cereus strain isolated from béchamel sauce. Two models, a quadratic polynomial equation and a reparameterized function, were assessed for their ability to describe the combined influence of aw and pH on the D85°C-values of the B. cereus isolate in tryptone soy broth. The performance of the models was validated by challenging the models with data independently collected in broth and béchamel sauce. Both models were found to adequately describe the validation data obtained in broth. However, it was determined that in béchamel sauce the predictions of the polynomial function not only showed bias (bias factor = 1.156) but were also fail-dangerous, as they deviated from the validation data by 17.2%. The reparameterized function was determined to be a good predictor of the D85°C-values in béchamel sauce as it showed no bias (bias factor = 1.033) and its predictions differed by only 7.9% from the validation data. The reparameterized function can be used to provide estimates of the minimum processing conditions required to achieve desired levels of spore inactivation within the aw and pH ranges studied and to determine the potential changes in heat resistance of B. cereus spores when aw and pH are changed, for example, during product reformulation. As validation of heat resistance models is rarely performed, let alone in actual food products, the models evaluated and validated in this study (in particular the reparameterized function) are of immediate relevance to the food industry. [ABSTRACT FROM AUTHOR]

Egg washing is currently not permitted within the European Union, with few exceptions. This is mainly because there are concerns that cuticle damage could occur during or after the washing process, as a result of a suboptimal operation. In this study, the cuticle coverage levels of 400 washed or unwashed eggs, derived from either a brown or a white egg-laying flock at the end of lay, were compared. The eggs from older hens inherently have poorer cuticle coverage and as a result arguably constitute a greater risk to consumer safety if they are then washed. Thus, the effects of the washing procedure used in this study on cuticle quality were tested under the worst-case scenario. A standard Swedish egg washing process was used. The cuticle coverage of the eggs was assessed by a colorimeter by quantifying the color difference before and after staining with Tartrazine and Green S. The cuticle of an additional 30 eggs from each of the four groups was then visually assessed by scanning electron microscopy. The staining characteristics of the cuticle varied greatly within each group of eggs and showed that the washing process did not lead to cuticle damage. Scanning electron microscopy confirmed that there was no irreversible damage to the cuticle of the washed eggs and that it was not possible to correctly assign the treatment (washed or not) based on a visual assessment. In conclusion, no evidence could be found to suggest that the washing procedure used in this investigation irreversibly changed the quality of the cuticle. [ABSTRACT FROM AUTHOR]

Abstract: Chitosan is a biopolymer with antimicrobial activity and film-forming properties. In this study, the effects on Salmonella shell contamination and trans-shell penetration of coating hens'' eggs with chitosan was evaluated. A chitosan was selected from eight types (four non-commercial and four commercial) based on its antimicrobial activity against Salmonella enterica serovar Enteritidis (S. Enteritidis). For this purpose, a contact plate method was developed and chitosans were applied at a concentration of 0.25% (w/v). A commercial type with a molecular weight of 310–375kDa and a deacetylation degree of 75% that reduced S. Enteritidis by 0.71log10 colony forming units compared to the control (without chitosan) was selected for further studies. The chitosan was shown to have antimicrobial activity against other egg borne bacteria, i.e., Acinetobacter baumannii, Alcaligenes sp., Carnobacterium sp., Pseudomonas sp., Serratia marcescens and Staphylococcus warneri, and against S. enterica serovar Typhimurium, Escherichia coli and Listeria monocytogenes. The effects of various concentrations of the selected chitosan (0.25%, 1% and 2%) on Salmonella shell contamination and trans-shell penetration were assessed using the agar molding technique. Effective reduction of eggshell contamination could not be demonstrated, but trans-shell penetration was significantly reduced in the presence of a 2% chitosan eggshell coating, with only 6.1% of the eggs being penetrated compared to 24.5% of the uncoated eggs. It was concluded that the 2% chitosan coating has the potential to reduce contamination of egg contents resulting from trans-shell penetration by S. Enteritidis. [Copyright &y& Elsevier]

Washing eggs in sterile plastic bags with diluent is an efficient sample preparation method for the determination of the bacterial contamination on eggshells. The total count of aerobic bacteria and the total count of Gram-negative bacteria on the eggshell can be used to detect critical contamination points in the egg production chain. The number of eggs to be sampled in a point of the production chain was determined on a statistical basis and fixed on 40 for non-graded eggs and on 20 for graded eggs. In two production chains, one cage production and one organic production system, critical contamination points were identified. The most critical point for the cage production system was the metal mat at the start of the grading, candling and packaging machine, for the organic production system it was the initial contamination in the nest boxes. With the exception of heavily soiled shells, like shells from eggs collected from the ground (ground eggs), there is a poor correlation between the level of bacterial contamination and the visual eggshell contamination. A positive correlation was found between the initial bacterial eggshell contamination and the concentration of bacteria in the air of the poultry houses. [Copyright &y& Elsevier]

• Vaccination of sows increased the concentration of maternally derived antibodies in 3-day old piglets. • Piglet and fattening pig vaccination resulted in higher sample-to-positive (S/P) ratios at slaughter compared to non-vaccinated pigs. • Piglet and fattening pig vaccination could interfere with serology-based Salmonella monitoring programs at slaughter. 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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Hygiene in animal production is key for both farm management and food safety. Cleaning and disinfection (C&D) of broiler houses is essential to manage farm hygiene. Still high levels of total aerobic flora after C&D in broiler houses are reported. However, little is known about the microbial composition after cleaning (AC) and after disinfection (AD). In addition, the question why some bacterial species/isolates are still present AD whereas others are killed remains. The study was carried out in 4 broiler houses. Sampling was performed AC and AD. The disinfectant was based on hydrogen peroxide and peracetic acid. Enumerations were carried out for total aerobic flora, Enterococcus spp. and Enterobacteriaceae. The dominant bacteria present was assessed by (GTG)5 analysis and 16S rRNA gene sequence analysis. Moreover, minimum bactericidal concentration (MBC) tests were carried out on 18 selected isolates belonging to the Enterobacteriaceae family and 10 Enterococcus faecium isolates. A wide variety of bacteria were detected AC and AD. In total, 363 and 255 isolates were identified AC and AD, respectively, resulting in a total of 109 identified species. The most dominant bacteria belonged to Brevibacterium, Brachybacterium, and Staphylococcus AC and Bacillus, Brevibacterium, and Staphylococcus AD. At both sampling moments, Enterococcus faecium was dominant among the Enterococcus spp. isolates. On the selective medium for Enterobacteriaceae, the genera Enterobacter and Pantoea and Aeromonas (non Enterobacteriaceae) were dominant AC while Escherichia, Lelliottia, and Pantoea were dominant AD. In addition, species pathogenic to poultry and humans were identified not only AC but also AD. MBC results showed no trend in selection of less susceptible isolates for the used disinfectant AD compared to AC. Finally, the recommended concentration of the disinfectant (i.e., 0.5% commercial solution of hydrogen peroxide and peracetic acid) seemed too low to kill Enterobacteriaceae. [ABSTRACT FROM AUTHOR]

A novel method for geometry generation of Representative Unit Cells (RUC) of textile composites is presented. The technique retains the advantage of an analytical formulation from industrial practice however introduces variable asymmetric yarn cross-sectional shapes and paths which can be fitted to the yarn shapes and cross-sectional areas as observed from in-situ measurements. In this way interpenetrations and incorrect fibre volume fractions, which occur when using idealized constant yarn cross sections for RUC generation, are avoided. Meshing becomes easier and no fibre volume corrections are required. The new technique is validated through a comparison of 1) the novel RUC to 2) an Idealized RUC with constant yarn cross section; and 3) a model constructed from direct in-situ micro computed X-ray tomographic measurements of a carbon-epoxy weave (In-situ Model). With all three models a reasonable agreement with experimentally obtained elastic properties is found. The stress predicted by the Idealized RUC is significantly different than predicted by the RUC generated with the new method and the In-situ Model. The latter two are in good agreement which indicates that the MESI RUC can be used for material strength prediction. The MESI RUC is also substantially less computationally intensive. Next to the construction of improved RUCs, the technique is an excellent alternative for advanced unit cell generation techniques based on production process simulations in the case that the production process is unknown or an analytic periodic geometry is required. [ABSTRACT FROM AUTHOR]

Bacterial chondronecrosis with osteomyelitis (BCO) is the most common cause of lameness in commercial broilers. Growing broilers on wire flooring provides an excellent experimental model for reproducibly triggering significant levels of lameness attributable to BCO. In the present study we evaluated the efficacy of adding HyD (25-OH vitamin D3) to the drinking water as a preventative/prophylactic treatment for lameness. Broiler chicks were reared on 5 x 10 ft flat wire floor panels within 6 environmental chambers. Three chambers were supplied with tap water (Control group) and the remaining chambers were supplied with HyD (HyD group: 0.06 mL HyD solution/L water; dosing based on the HyD Solution label to provide 33.9 μg 25-OHD3/L) from d 1 through 56. Feed was provided ad libitum and was formulated to meet or exceed minimum standards for all ingredients, including 5,500 IU vitamin D3/kg. Lameness initially was detected on d 28, and the cumulative incidence of lameness on d 56 was higher in the Control group than in the HyD group (34.7 vs. 22.7%, respectively; P = 0.03; Z-test of proportions; chambers pooled). The most prevalent diagnoses for lame birds were osteochondrosis and osteomyelitis (BCO) of the proximal femora (52%) and tibiae (79%), accompanied by minor incidences of tibial dyschondroplasia (0.33%), spondylolisthesis, or kinky back (0.67%), and twisted legs (1%). Broilers that survived to d 56 without developing lameness did not differ in BW when compared by group within a gender. The wire flooring model imposes a rigorous, sustained challenge that undoubtedly is much more severe than typically would be experienced by broilers under normal commercial conditions. Therefore the encouraging response to HyD supplementation in the present study supports the potential for 25-OH vitamin D3 to attenuate outbreaks of lameness caused by BCO in commercial broiler flocks. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Highlights: [•] S. Gallinarum outbreak strains and the SG9R vaccine have identical DNA fingerprints. [•] Whole-genome sequencing shows 9 SNPs distinguishing an outbreak strain from SG9R. [•] Mutations in the aceE and rfaJ genes could explain the reversion to virulence of SG9R. [Copyright &y& Elsevier]

Reducing Campylobacter shedding on the farm could result in a reduction of the number of hu-man campylobacteriosis cases. In this study, we first investigated if allicin, allyl disulfide, and garlic oil ex-tract were able to either prevent C. jejuni growth or kill C. jejuni in vitro. Allyl disulfide and garlic oil extract reduced C. jejuni numbers in vitro below a detectable level at a concentration of 50 mg/kg (no lower concen-trations were tested), whereas allicin reduced C. jejuni numbers below a detectable level at a concentration as low as 7.5 mg/kg. In further experiments we screened for the anti- C. jejuni activity of allicin in a fermentation system closely mimicking the broiler cecal environment using cecal microbiota and mucus isolated from C. jeju-ni-free broilers. During these fermentation experiments, allicin reduced C. jejuni numbers below a detectable level after 24 h at a concentration of 50 mg/kg. In con-trast, 25 mg/kg of allicin killed C. jejuni in the first 28 h of incubation, but anti-C. jejuni activity was lost af-ter 48 h of incubation, probably due to the presence of mucin in the growth medium. This had been confirmed in fermentation experiments in the presence of broiler cecal mucus. Based on these results, we performed an in vivo experiment to assess the prevention or reduction of cecal C. jejuni colonization in broiler chickens when allicin was added to drinking water. We demonstrated that allicin in drinking water did not have a statisti-cally significant effect on cecal C. jejuni colonization in broilers. It was assumed, based on in vitro experiments, that the activity of allicin was thwarted by the presence of mucin-containing mucus. Despite promising in vitro results, allicin was not capable of statistically influenc-ing C. jejuni colonization in a broiler flock, although a trend toward lower cecal C. jejuni numbers in allicin-treated broilers was observed. [ABSTRACT FROM AUTHOR]

Campylobacter is the most commonly re-ported gastrointestinal bacterial pathogen in humans in many developed countries. During slaughter of broiler flocks, it is difficult to avoid contamination of broiler carcasses. This study aimed to quantify Campylobacter contamination on broiler carcasses at 5 points in the slaughter processing during the slaughter of a Cam-pylobacter-colonized flock by real-time PCR and con-ventional enumeration. In addition, the decontamina-tion effect of neutral electrolyzed oxidizing (EO) water and 1.5% lactic acid (pH 2.0) were evaluated. During processing, the Campylobacter counts on the carcasses declined toward the end of the processing line. The log counts on the carcasses as determined by quantita-tive real-time PCR (qPCR), decreased from 9.37 after scalding to 8.08 after the last cooling step. Enumera-tion of the Campylobacters on plates revealed the same trend, although the counts per carcass were generally 3 logs lower. After scalding, a mean of 6.86 log cfu/car-cass were counted, which decreased to 4.83 log cfu/car-cass after the last cooling step. Submerging carcasses after scalding in EO water gave a significant reduction of 1.31 log cfu/carcass by enumeration on plates and a not significant reduction of 0.53 log cfu/carcass by qPCR. Treatment of the carcasses after the inside-out-side bird washer led to reductions from 0.09 to 0.91 log cfu/carcass, although not significant. After submerging the carcasses in a 1.5% lactic acid solution, significant reductions of 1.62 and 1.24 log cfu/carcass by qPCR and enumeration, respectively, were observed. Spray-ing the carcasses with lactic acid led to nonsignificant reductions of 0.68 log cfu/carcass determined by qPCR and 0.26 log cfu/carcass by enumeration. Both EO wa-ter and lactic acid seem promising for implementation in poultry processing plants. [ABSTRACT FROM AUTHOR]

Bacterial gastroenteritis caused by ther-motolerant Campylobacter species, mainly Campy-lobacter jejuni, has been the most reported zoonotic disease in many developed countries in recent years. Reducing Campylobacter shedding on the farm could result in a reduction of the number of campylobacterio-sis cases. In 2 independent broiler seeder experiments, in which broiler chickens were orally inoculated with 2 amounts of Enterococcus faecalis MB 5259, we es-tablished whether a live E. faecalis strain was capable of reducing cecal Campylobacter colonization in broiler chickens. In previous in vitro experiments it has been demonstrated that this E. faecalis MB 5259 displays anti- Campylobacter activity. The effect of pH and bile salts on E. faecalis MB 5259 showed that growth and survival of E. faecalis MB 5259 can be impaired dur-ing passage through the gastrointestinal tract of broiler chickens. Despite these results E. faecalis MB 5259 was capable of colonizing the broiler ceca. Contrary to the in vitro experiments, in which E. faecalis MB 5259 in-hibited C. jejuni MB 4185 growth, no inhibition was observed in the in vivo experiments independent of the inoculum size. [ABSTRACT FROM AUTHOR]

The aim of this study was to investigate whether the main coagulase-negative staphylococcus (CNS) species involved in bovine intramammary infections (IMI) possess specific characteristics that promote colonization of the udder. Virulence markers associated with biofilm formation, antimicrobial resistance, and biocide tolerance were compared between typically contagious CNS species (Staphylococcus chromogenes, Staphylococcus epidermidis, Staphylococcus haemolyticus, and Staphylococcus simulans) and those rarely causing IMI (Staphylococcus sciuri, Staphylococcus equorum, and others) to find possible associations with pathogenicity. Coagulase-negative staphylococcus isolates (n = 366) belonging to 22 different species were analyzed by PCR for the presence of the biofilm-associated genes bap and icaA, and the methicillin resistance gene mecA. A selection of 82 isolates was additionally tested for their susceptibility to 5 antibiotics and 2 commercial teat dip products. Minimum inhibitory concentrations of antimicrobials were determined by Etest (AB bio-Mérieux, Marcy l'Etoile, France), and a microdilution method was optimized to determine minimum biocidal concentrations of teat dips. The bap, icaA, and mecA genes were detected significantly more in isolates from CNS species typically living in the cows' environment than in isolates from IMI-causing species. Antimicrobial resistance was mainly against erythromycin (23%) or oxacillin (16%), and was detected more often in the environmental species. The isolates least susceptible to the teat dips belonged to the IMI-causing species Staph. chromogenes and Staph. simulans. We concluded that carriage of biofilm genes and antimicrobial resistance were not associated with the ability to colonize the mammary gland because free-living CNS species constituted a more significant reservoir of biofilm and resistance determinants than did IMI-causing species. In contrast, increased tolerance to biocides may favor the establishment of bovine IMI by some CNS species. [ABSTRACT FROM AUTHOR]

Since 2007, a national Salmonella control program including obligatory vaccination has been on-going in Belgium. In this context, the aim of the pres-ent study was to investigate the diversity of Salmonella enterica serovar Enteritidis isolates on 5 persistently contaminated Belgian layer farms and to examine the potential sources and transmission routes of Salmonella Enteritidis contamination on the farms during succes-sive laying rounds. A collection of 346 Salmonella iso-lates originating from the sampled farms were char-acterized using a combination of multilocus variable number of tandem repeat analysis (MLVA) and phage typing (PT). On each farm, one or 2 dominant MLVA-PT types were found during successive laying cycles. The dominant MLVA type was different for each of the individual farms, but some farms shared the same dominant phage type. Isolates recovered from hens' feces and ceca, egg contents, eggshells, vermin (mice, rats, red mites, and flies), and pets (dog and cat feces) had the same MLVA-PT type also found in the inside henhouse environment of the respective layer farm. Per-sistent types were identified in the layer farm inside environment (henhouse and egg collecting area). Fur-thermore, this study demonstrated cross-contamination of Salmonella between henhouses and between the hen-house and the egg collecting area. Additional isolates with a different MLVA-PT type were also recovered, mainly from the egg collecting area. A potential risk for cross-contamination of Salmonella between the indi-vidual layer farms and their egg trader was identified. [ABSTRACT FROM AUTHOR]

Campylobacteriosis is the most reported bacterial-mediated gastroenteritic disease in many developed countries. Broiler chickens are a natural host for Campylobacter spp., and contaminated poultry meat products are a major source for transmitting pathogenic Campylobacter strains to humans. Currently, no intervention measure efficiently and effectively controls this pathogen in poultry flocks. Medium-chain fatty acids (caproic, caprylic, capric, and lauric acids) show a marked anti-Campylobacter activity in vitro. However, in recent trials using our in vivo models, administering these acids to the feed of broiler chicks neither prevented nor reduced cecal C. jejuni colonization in broilers. In the present study, we examined whether a drinking water application of medium-chain fatty acids might be more effective in combating Campylobacter colonization in poultry. Although Campylobacter colonization and transmission was not reduced, we demonstrate that adding an emulsion of a mixture of caproic, caprylic, capric, and lauric acids to the drinking water of broiler chicks reduces their colonization susceptibility and prevents C. jejuni survival in drinking water. Thus, the merit of water applications of medium-chain fatty acids is the reduction of the probability of Campylobacter entry into and transmission throughout a flock. [ABSTRACT FROM AUTHOR]

The aim of this study was to closely examine the Salmonella enterica serovar Enteritidis environmental contamination on persistently positive layer farms in Belgium during successive laying cycles. All of the farms were required to vaccinate their layers under the national control program for Salmonella. Seven farms with previous or current Salmonella Enteritidis contamination were monitored during different stages of the laying period and after cleaning and disinfection (CD). Environmental samples, including from the equipment and vermin, were taken in the henhouse and egg-collecting area. Dilutions were performed to define the degree of Salmonella Enteritidis contamination. Eggshells, egg contents, and ceca were also tested for Salmonella. At the end of the first sampled laying period, 41.6% of the environmental samples were contaminated with Salmonella Enteritidis. After CD, the prevalence dropped to 11.4%. On average, the prevalence in the second laying period increased again: 17.8, 18.4, and 22.3% at the onset, middle, and end of the lay period, respectively. After CD before the third laying period, the prevalence decreased to 6.6% and stabilized at the onset of lay (6.3%). During lay, as well as after CD, a wide variety of contaminated environmental samples were found; for example, in the henhouse, in the egg-collecting area, on mobile equipment and in or on vermin. In the henhouse during laying, the most recurrent and highly contaminated sites were the overshoes, floor, manure belt, and hen feces. The egg-collecting area had a significantly higher number of contaminated samples compared with that of the henhouse. For both sites, the floor appeared to be the most suitable sampling site to estimate the Salmonella Enteritidis status of the farms. Eggshell and egg content contamination varied between 0.18 and 1.8% and between 0.04 and 0.4%, respectively. In total, 2.2% of the analyzed ceca contained Salmonella Enteritidis. This study revealed that Salmonella Enteritidis is present in the environment of persistently Salmonella Enteritidis-contaminated layer farms, demonstrated that in many cases Salmonella Enteritidis contamination was not eliminated after CD, and identified the egg-collecting area as a critical point on most farms. [ABSTRACT FROM AUTHOR]

The present study evaluated Escherichia coli, Enterococcus faecalis, and Enterococcus hirae as potential indicator organisms for the possible Salmonella Enteritidis (SE) presence in layer farms after cleaning and disinfection by comparing their susceptibility to disinfection. A quantitative suspension disinfection test according to European Standard EN 1656 was performed using disinfection products CID20 and Virocid (both from CID Lines, leper, Belgium). In a preliminary test, the sensitivity to both disinfection products was compared between ATCC strains of SE, E, coli, En, faecalis, and En, hirae. The sensitivity of SE to disinfection was most comparable to that of E. coli, A second disinfection test compared the elimination of E, coli to SE ATCC strains as well as field strains. Results showed no significant effect regarding the strain (P > 0.05 for CID20 and Virocid), meaning that no difference was detected in sensitivity toward disinfection. When comparing the sensitivity in general at species level for all concentrations of disinfectant used, no significant difference was found between E, coli and SE in sensitivity to Virocid {P > 0.05). In conclusion, because of its similar response to disinfection in a suspension disinfection test, E, coli could be used as an indicator for possible Salmonella presence after cleaning and disinfection. [ABSTRACT FROM AUTHOR]

Camp ylobacter jejuni is the most common cause of bacterial-mediated diarrheal disease worldwide. Because poultry and poultry products are a major source of C. jejuni infections in humans, efforts should be taken to develop strategies to decrease Camp ylobacter shedding during primary production. For this purpose, the efficacy of medium-chain fatty acids (MCFA) as feed additives to control C. jejuni colonization in broiler chickens was analyzed. First, the antimicrobial activity of the MCFA caproic, caprylic, and capric acid on C. jejuni was evaluated in vitro. Minimal inhibitory concentrations were 0.25 mM for caproic and 0.5 mM for caprylic and capric acids at pH 6.0 and 4 mM for all 3 compounds at pH 7.5. Time-kill curves revealed strong bactericidal properties of the tested compounds toward C. jejuni at pH 6.0. Concentrations as low as 4 mM caprylic and capric acids and 16 mM caproic acid killed all bacteria within 24 h. Capric acid had the highest activity, with concentrations of 4 mM killing all bacteria within the hour. Together these data show a profound bactericidal, dose-dependent activity of the tested MCFA toward C. jejuni in vitro. For this reason, the effect of these 3 MCFA on C. jejuni was evaluated in vivo. The addition of any of the acids to the feed, from 3 d before euthanization, was not capable of reducing cecal Camp ylobacter colonization in 27-d-old broilers experimentally infected with C. jejuni at 15 d of age. Using a cecal loop model, sodium caprate was not able to reduce cecal Camp ylobacter counts. When time-kill curves were conducted in the presence of chick intestinal mucus, capric acid was less active against C. jejuni. At 4 mM, all bacteria were killed only after 24 h. Thus, despite the marked bactericidal effect of MCFA in vitro, supplementing these acids to the feed does not reduce cecal Camp ylobacter colonization in broiler chickens under the applied test conditions, probably due to the protective effect of the mucus layer. [ABSTRACT FROM AUTHOR]

For laying hens, the effects of housing system on bacterial eggshell contamination and eggshell quality is almost exclusively studied in experimental hen houses. The aim of this study was to compare eggshell hygiene and quality under commercial conditions. Six flocks of laying hens in furnished cages and 7 flocks in noncage systems were visited when hens were about 60 wk of age. Farms from Belgium, the Netherlands, and Germany were included in the study. The following parameters were determined on eggs sampled at the egg belts: 1) bacterial eggshell contamination, as expressed by total count of aerobic bacteria and number of Enterobacteriaceae; 2) proportion of dirty eggs; and 3) proportion of cracked eggs and eggs with microcracks. Considerable within-flock differences were found in eggshell contamination with total count of aerobic bacteria, both for furnished cages (P ⩽ 0.001, range 4.24 to 5.22 log cfu/eggshell) and noncage systems (P ⩽ 0.001, range 4.35 to 5.51 log cfu/eggshell). On average, lower levels of contamination with total count of aerobic bacteria (4.75 vs. 4.98 log cfu/eggshell; P ⩽ 0.001) were found on eggshells from furnished cages compared with noncage systems. Concerning Enterobacteriaceae, no significant difference in average eggshell contamination between both systems could be shown. The total percentage of cracked eggs was higher (P < 0.01) in furnished cages (7.8%) compared with noncage systems (4.1%). This was, however, due to the high percentage of cracked eggs (24%) observed on one of the furnished cage farms. We conclude that bacteriological eggshell contamination and percentage of cracked eggs differed substantially between individual farms using the same housing system. This may also explain some discrepancies between the findings of the present study versus some findings of previous experimental studies or studies on a small number of farms. Although statistically significant, the average differences in bacteriological contamination of nest eggs between both housing systems have limited microbiological relevancy. [ABSTRACT FROM AUTHOR]

Abstract: In this paper the selection of potential probiotic strains for producing fermented liquid feed (FLF) for pigs is presented. One hundred and forty six strains of lactic acid bacteria were isolated from ileal digesta of piglets, cecal digesta of slaughter pigs and FLF from a preliminary trial at our laboratory. The strains were identified with repetitive-element PCR and partial 16S rRNA sequencing and screened for acidification potential. Ten of the strains were selected and their culture supernatant was checked for antibacterial activity against Salmonella spp. Also, they were tested in vitro in batch fermentations at 30°C for 72h, for their capacity to produce FLF (water:feed ratio of 4:1) with a concentration of acetic acid less than 40mmol/L and of lactic acid above 100mmol/L, resulting in a pH below 4.5. The Lactobacillus strains MB 3123 (L. johnsonii), MB 3182 (L. salivarius group) and MB 3083 (L. plantarum) were found to be very effective for the production of FLF. The FLF produced by including this strain had a pH drop from 5.9 to 4.2, 4.3 and 4.4 after 24h and to 4.1 after 48h of fermentation. Also, these three strains showed high antimicrobial activity against Salmonella spp. and produced more than 100mmol/L lactic acid after 48h fermentation. The MB 3170 strain increased the lactic acid concentration above 100mmol/L after 48h, but at 24h the pH was only 4.5. The results indicate that it is possible to screen for probiotic strains capable of fermenting feeds and of producing high quantities of lactic acid with possible antimicrobial effects in the gastro-intestinal tract of pigs. [Copyright &y& Elsevier]

Abstract: Trans-shell infection routes and whole egg contamination of 7 selected bacterial strains; Staphylococcus warneri, Acinetobacter baumannii, Alcaligenes sp., Serratia marcescens, Carnobacterium sp., Pseudomonas sp. and Salmonella enteritidis, recovered from egg contents, were studied. The first objective was to correlate bacterial eggshell penetration with various eggshell characteristics and bacterial strains. An agar approach was used to assess the eggshell penetration. The second objective was to assess the contamination of whole eggs with the bacterial strains; whole intact eggs were used in this case. The intact shells of agar-filled and whole eggs were inoculated with 103–104 cfu of the selected strains. During 3 weeks storage at 20 °C and 60% relative humidity, the bacterial eggshell penetration was regularly monitored. The whole egg contamination was only analyzed after 3 weeks. The eggshell characteristics such as area eggshell, shell thickness and number of pores did not influence the bacterial eggshell penetration. For each individual bacterial strain the mean cuticle deposition was lower for penetrated compared to non-penetrated eggshells. For the individual strain Carnobacterium sp. and for the global results of all strains this difference was statistical significantly. The whole egg contamination was not influenced by neither the area of the eggshell nor the porosity of the eggshell. The results of the agar approach indicate that the Gram-negative, motile and non-clustering bacteria penetrated the eggshell most frequently; Pseudomonas sp. (60%) and Alcaligenes sp. (58%) were primary invaders followed by S. enteritidis (43%). All selected strains were able to penetrate; penetration was observed most frequently after ca. 4–5 days. Particularly S. enteritidis was a primary invader of whole eggs: the membranes and/or the content of 32% of the whole eggs was contaminated. The remaining bacterial eggshell contamination with the selected strain was determined after 3 weeks storage. Penetrated eggshells and contaminated whole eggs showed a significantly higher bacterial contamination on the eggshell compared to non-penetrated eggshells and non-contaminated whole eggs respectively (global results of all strains). The influence of hen age on bacterial eggshell penetration and egg content contamination was not significant. While the agar approach is suitable to study the influence of the eggshell characteristics on the bacterial eggshell penetration, the intact egg approach gives an estimation of the penetration of the shell followed by the probability of survival and migration in whole eggs. [Copyright &y& Elsevier]