Bacterial contamination of table eggs and the influence of housing systems

With the introduction of alternative housing systems for laying hens in the EU, recent research has focussed on the bacterial contamination of table eggs, e.g. eggshell and egg content contamination. Contamination of eggshells with aerobic bacteria is generally higher for nest eggs from non-cage systems compared to nest eggs from furnished cages or eggs from conventional cages. Studies indicate limited or no systematic differences in eggshell contamination with aerobic bacteria between eggs laid in the nest boxes of furnished cages and eggs laid in conventional cages. The major differences found in experimental studies between cage- and non-cage systems are less pronounced under commercial conditions. The effect of housing system on eggshell contamination with specific groups of bacteria is variable. Limited information is available on the influence of housing system on egg content contamination. Recent research does not indicate large differences in egg content contamination between eggs from cage- and non-cage systems (ignoring outside nest and floor eggs). The microflora of the eggshell is dominated by Gram-positive bacteria, whereas Gram-negative bacteria are best equipped to overcome the antimicrobial defences of the egg content. Much of the research on eggshell and egg content contamination focuses on Salmonella, since infection with Salmonella enteritidis, resulting from the consumption of contaminated eggs or egg products, is still a major health problem. Observed Salmonella prevalence on the eggshell and in the egg content vary, depending on the fact whether investigations were based on randomly sampled table eggs or on eggs from naturally infected hens. The limited information available on other pathogens shows that they are exclusively isolated from the eggshell and not from the internal contents

Strategies for estimating human exposure to mycotoxins via food

In this review, five strategies to estimate mycotoxin exposure of a (sub-) population via food, including data collection, are discussed with the aim to identify the added values and limitations of each strategy for risk assessment of these chemicals. The well-established point estimate, observed individual mean, probabilistic and duplicate diet strategies are addressed, as well as the emerging human biomonitoring strategy. All five exposure assessment strategies allow the estimation of chronic (long-term) exposure to mycotoxins, and, with the exception of the observed individual mean strategy, also acute (short-term) exposure. Methods for data collection, i.e. food consumption surveys, food monitoring studies and total diet studies are discussed. In food monitoring studies, the driving force is often enforcement of legal limits, and, consequently, data are often generated with relatively high limits of quantification and targeted at products suspected to contain mycotoxin levels above these legal limits. Total diet studies provide a solid base for chronic exposure assessments since they provide mycotoxin levels in food based on well-defined samples and including the effect of food preparation. Duplicate diet studies and human biomonitoring studies reveal the actual exposure but often involve a restricted group of human volunteers and a limited time period. Human biomonitoring studies may also include exposure to mycotoxins from other sources than food, and exposure to modified mycotoxins that may not be detected with current analytical methods. Low limits of quantification are required for analytical methods applied for data collection to avoid large uncertainties in the exposure due to high numbers of left censored data, i.e. with levels below the limit of quantification

Genomic and toxigenic heterogeneity of bacillus cereus sensu lato Isolated from ready-to-eat foods and powdered milk in day care centers in Colombia

Bacillus cereus sensu lato (s.l.) is a group of bacteria commonly found in diverse environments, including foods, with potential to cause emesis and diarrhea. In Colombia, it is one of the main foodborne pathogens. The aim of this study was to determine the genomic and toxigenic heterogeneity of B. cereus s.l. isolated from ready-to-eat foods and powdered milk collected in day care centers of Medellin, Colombia. Of 112 B. cereus s.l. isolates obtained, 94% were beta-hemolytic. Toxigenic heterogeneity was established by the presence of nheABC, hblCDAB, cytK2, entFM, and cesB toxigenic genes. The nheABC operon and entFM gene were most frequently detected in the isolates, whereas the cesB gene was not found. According to the toxin genes content, nine toxigenic profiles were identified. A 44% of isolates had profiles with all genes for nonhemolytic enterotoxin, hemolysin BL, and enterotoxin FM production (profiles II and IV). Pulsed-field gel electrophoresis analysis indicated a high genomic heterogeneity among the B. cereus s.l., with 68 isolates grouping into 16 clusters and 33 placed separately in the dendrogram. This study provides useful information on the safety of ready-to-eat foods and powdered milk in day care centers where children, a susceptible population, are exposed and it should incentive for more studies to understand the distribution of different toxin-encoding genes among B. cereus s.l. isolates, enabling detailed risk assessment

Inhibition of Salmonella Typhimurium by medium chain fatty acids in an in vitro simulation of the porcine caecum

To lower the contamination of pork meat with Salmonella, feed additives such as medium chain fatty acids (MCFA\u27s) can be applied at the primary production level. An in vitro continuous culture system, simulating the porcine caecum, was developed for investigating the effect of MCFAs on the pig intestinal microbial community. The system was monitored by plating on selective media, 16S rDNA PCR denaturing gradient gel electrophoresis (PCR-DGGE) and HPLC analysis of fermentation products. In a simulation of the porcine caecum without MCFA treatment, with Salmonella Typhimurium added after stabilization of the microbial community, the strain could establish itself at a stable population size of about 5 log cfu/ml. The effect of selected MCFAs was observed from all monitored parameters and depended on chain length and concentration applied. At a dose of 15 mM, caproate and caprinate did not show any pronounced effect, while a clear Salmonella inhibiting effect (3 log units reduction) was found for caprylate. Doubling the caprylate dose did not result in enhanced Salmonella inhibition

Expression study by real-time quantitative RT-PCR of the Salmonella typhimurium mntH gene

The aim of our study was to compare the mntH expression of different Salmonella Typhimurium strains and other Salmonella serovars with real-time RT-PCR. Following the mntH expression in function of the growth showed that the mntH expression of S. Typhimurium is growth dependent. A strong decrease of the mntH expression is noticed when the growth reaches 1.78 108 CFU/ml. After induction with EDTA or H2O2, variations between different S. Typhimurium strains were observed. For some S. Typhimurium strains a 10 to 20 times higher mntH expression was noticed after H2O2 induction. The EDTA induction was for most strains lower (5 to 10 times) but also variations between different strains were observed. The other Salmonella serovars were strongly induced after H2O2 but not after EDTA induction

Is competitive exclusion a valuable alternative for classical cleaning and disinfection of pig-growing units?

Colonization of the environment of pig-growing units by pathogenic microrganisms is an important factor in development of endemic diseases in pigs and, in spreading of zoonotic diseases. These pathogens are mostly controlled by the use of antibiotics and disinfection during vacancy. Because, the past years an increasing resistance against these measures is noticed, alternative methods such as competitive exclusion (CE) are promoted as promising. In this study the effect of a CE protocol on the bacterial infection in piggrowing units was compared to a classical cleaning and disinfection (C&D) protocol. Tests were performed during three successive production rounds using multiple identical pig-growing units. CE protocol consisted of cleaning (no disinfection) after loading piglets and spraying probiotic bacteria (Bacillus spp. spores) during vacancy and production. The cleaning product also contained Bacillus spores. Sampling was performed at different time-points: immediately after pig loading (manure still present); 24 hours after cleaning (CE units) or after disinfection (control units); after one week and five weeks of production (piglets present). At each time point, swab samples for analyses were taken. Enumerations of bacterial spores, Enterococcus spp., E. coli, fecal coliforms and MRSA and detections of E. coli, fecal coliforms and MRSA were performed. Next to bacterial analyses, also feed conversion and fecal consistency was monitored. This study showed that, although probiotic spores were administered well, the analyzed bacteria were not decreased after three production rounds in CE units and remained on the same level as the control units (C&D). Also, the infection pressure in CE units during vacancy was not as much reduced as after the disinfection-step in control units. Finally, no differences in feed conversion and fecal consistency were found. These results indicate that the used CE protocol is not a valuable alternative for classical C&D

The prevalence of Salmonella, Campylobacter and VTEC in pig farms

Four different pig farms were sampled for the prevalence of Salmonella, Campylobacter and VTEC. From a total of 215 rectal samples of individual pigs, 15 rectal samples taken from animals at the same farm were positive for Salmonella. The Salmonella status of the pigs at this farm differed from one age group to another. S. Typhimurium was isolated from all the rectal samples and S. Typhimurium and S. Schwarzengrund were isolated from the environment. On two other farms Salmonella was only present in the environment with S. London and S. Typhimurium as serotypes. With cut-off value %OD\u3e 40 in the ELISA we found a good correlation with the Salmonella status of the farm. The presence of Campylobacter was tested in 150 rectal swabs, 51 of these, spread over the four farms, turned out positive. All the strains were identified as Campylobacter coli by a species-specific PCR. To determine if pigs are a reservoir of VTEC a total of 289 samples were screened for the presence of VTEC and 54 strains were isolated that each carried one virulence gene. Thirty-one strains carried the vt2e variant of the vt2 gene, four strains harboured the hly A gene and 19 the eaeA gene

Interventional Cardiology in Europe 1993

An annual survey on cardiac interventions in Europe is performed by the working group on Coronary Circulation of the European Society of Cardiology with the help of the national societies of cardiology. A questionnaire about cardiac interventions in 1993 was mailed to a representative of the national societies of 35 members of the European Society of Cardiology. The data collection of coronary interventions was delayed by slow backreporting and from 10 of the 35 national members data were missing or grossly incomplete. They were excluded from the analysis. Coronary anglography A total of 756 822 coronary angiograms were reported resulting in an incidence of 1146 ± 1024 per 106 inhabitants, ranging from 24 (Romania) to 3499 (Germany). This represents an increase of 12% compared to 1992. Germany (279 882 cases), France (157 237), the United Kingdom (77 000), Italy (44 934) and Spain (37 591) registered 79% of all the coronary angiograms performed. Percutaneous transluminal coronary angioptasty A total of 183 728 percutaneous transluminal coronary angioplasty cases were reported in 1993, 24% more than in 1992. On average, they accounted for 18 ± 7% (range 8 (Romania) to 35% (Sweden)) of the coronary angiograms. Most of these percutaneous transluminal coronary angioplasties (82%) were confined to a single vessel. In 13% only, percutaneous transluminal coronary angioplasty took place immediately after the diagnostic study. Adjusted per capita, Germany ranks first with 873 percutaneous transluminal coronary angioplasties per 106 inhabitants, followed by France (737), Holland (725), Belgium (713), and Switzerland (665). The European mean of percutaneous transluminal coronary angioplasties per 106 inhabitants was 270 ± 279, representing an increase of 14% compared with 1992. A major in-hospital complication was reported in 3.8% of the patients undergoing percutaneous transluminal coronary angioplasty: 0.6% hospital deaths, 1.5% emergency coronary artery bypass graftings, and 1.7% myocardial infarctions. New devices In 1993 stents were implanted in 6444 patients (3.5% of all percutaneous transluminal coronary angioplasty patients), equally distributed between bail-out situations (53%) and elective procedures. The 14 stent implanting countries showed a mean increase in the incidence of coronary stenting of 53% compared with 1992. Other interventional devices were applied in 7045 cases, i.e. 3.8% of all percutaneous transluminal coronary angioplasty cases. Coronary ultrasound (2194 cases) and coronary angioscopy (380 cases) were performed infrequently. Non-coronary interventions Valvuloplasties were the most frequently performed non-coronary interventions. Six European countries performed more than 300 valvuloplasties each in 1993. Most of them were mitral valvuloplasties in southern countries. Conclusions Although partial backreporting might bias conclusions, several findings of this survey are noteworthy for the participating countries: The number of percutaneous transluminal coronary angioplasties is universally increasing. There is an extremely wide range of coronary angiography and percutaneous transluminal coronary angioplasties performed per population. The most common additional procedure is a stent implantation while other new devices are only rarely applied. Mitral valvuloplasty is the most frequently performed non-coronary intervention. (Eur Heart J 1996; 17: 1318-1328

SPI-2 of Salmonella Typhimurium is not necessary for long term colonization of pigs

Unravelling the role of Salmonella virulence factors in the porcine host could greatly contribute to the development of control measures such as vaccination. The virulence genes located on the Salmonella Pathogenicity Island 2 (SPI-2) are indispensable for the induction of systemic disease and persistence in BALB/c mice. The role of this pathogenicity island in the pathogenesis of Salmonella Typhimurium infections in pigs is not documented. Therefore, in the present study, the interactions of a porcine field strain of Salmonella Typhimurium and a non-polar isogenic SPI-2 (D-ssrA) deletion mutant were compared in both in vitro and in vivo models. The ssrA mutant strain displayed decreased SPI-2 expression levels in vitro and was attenuated in a mouse model after oral inoculation. No difference was seen in the expression of SPI-1 related virulence genes. Through flowcytometric analysis, the ssrA mutant strain was found to be moderately attenuated in intracellular replication in porcine macrophages in vitro. In an infection experiment, 2 groups of 10 piglets were orally inoculated with the wild type or the ssrA mutant strain. The infection of the animals inoculated with the ssrA mutant strain followed a similar course as the animals infected with the wild type strain. At days 5 and 28 post inoculation, the animals of both groups were infected to the same extent in the gut and gut-associated lymphoid tissue, as well as in the mternal organs. These results suggest that SPI-2 of Salmonella Typhimurium may not contribute to the colonization of pigs to the same extent as it contributes to the colonization of BALB/c mice