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Biomedical subjects

L Herman

Publications and source records attributed to L Herman.

At least 19 recordsLinked to original sources

Quantitative risk assessment of Campylobacter spp. in poultry based meat preparations as one of the factors to support the development of risk-based microbiological criteria in Belgium.

The objective of this study was to do an exercise in risk assessment on Campylobacter spp. for poultry based meat preparations in Belgium. This risk assessment was undertaken on the demand of the competent national authorities as one of the supportive factors to define risk-based microbiological criteria. The quantitative risk assessment model follows a retail to table approach and is divided in different modules. The contamination of raw chicken meat products (CMPs) was represented by a normal distribution of the natural logarithm of the concentration of Campylobacter spp. (ln[Camp]) in raw CMPs based on data from surveillance programs in Belgium. To analyse the relative impact of reducing the risk of campylobacteriosis associated with a decrease in the Campylobacter contamination level in these types of food products, the model was run for different means and standard deviations of the normal distribution of the ln[Camp] in raw CMPs. The limitation in data for the local situation in Belgium and on this particular product and more precisely the semi-quantitative nature of concentration of Campylobacter spp. due to presence/absence testing, was identified as an important information gap. Also the knowledge on the dose-response relationship of Campylobacter spp. was limited, and therefore three different approaches of dose-response modelling were compared. Two approaches (1 and 2), derived from the same study, showed that the reduction of the mean of the distribution representing the ln[Camp] in raw CMPs is the best approach to reduce the risk of Campylobacter spp. in CMPs. However, for the simulated exposure and approach 3 it was observed that the reduction of the standard deviation is the most appropriate technique to lower the risk of campylobacteriosis. Since the dose-response models used in approach 1 and 2 are based on limited data and the reduction of the mean corresponds with a complete shift of the contamination level of raw CMPs, demanding high efforts from the poultry industry, it is proposed to lower the standard deviation of the concentration of Campylobacter spp. in raw CMPs. This proposal corresponds with the elimination of the products that are highly contaminated. Simulation showed that eating raw chicken meat products can give rise to exposures that are 10(10) times higher than when the product is heated, indicating that campaigns are important to inform consumers about the necessity of an appropriate heat treatment of these type of food products.

Animals↗

Eggshell factors influencing eggshell penetration and whole egg contamination by different bacteria, including Salmonella enteritidis.

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 10(3) -10(4) cfu of the selected strains. During 3 weeks storage at 20 degrees 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.

Animals↗

Bacterial shell contamination in the egg collection chains of different housing systems for laying hens.

The bacterial eggshell contamination of eating eggs in different commercial housing systems; two conventional cages, one organic aviary system and one barn production, were compared. The total counts of aerobic bacteria and the total counts of Gram-negative bacteria on the shell were used to detect key points where contamination occurred and to study the progress of contamination in the egg collection and transportation chains. The key points in the chain were those where eggs accumulated on a short conveyor belt, initial shell contamination in the alternative housing systems and extra nest-boxes placed on the ground. The high bacterial load of floor eggs (>6.3 log CFU total aerobic flora/eggshell) explains why they cannot be used for eating. On average higher initial shell contamination with total counts of aerobic bacteria was found for eggs from the alternative housing systems compared to the conventional systems; respectively 5.46 compared to 5.08 log CFU/eggshell. However, initial contamination with total counts of Gram-negative bacteria on the shells was less in the alternative systems: 3.31 compared to 3.85 log CFU/shell. Initial bacterial shell contamination tended to correlate positively with the concentration of bacteria in the air of the poultry houses. Storing shell eggs, whether temporarily refrigerated or not, for 9 d or more, resulted in a decrease in bacterial eggshell contamination for both bacterial variables.

Air Microbiology↗

Eggshell penetration of hen's eggs by Salmonella enterica serovar Enteritidis upon various storage conditions.

1. The survival and penetration of Salmonella enterica serovar Enteritidis (SE) inoculated on the eggshell was examined upon storage for up to 20 d at real-life conditions (15 to 25 degrees C and 45 to 75% relative humidity (RH)). 2. Penetration was assessed by emptying the egg contents and filling the eggs with a selective medium that allowed visualising Salmonella growth on the inside of the shell and membrane complex. 3. The study of survival on the eggshells was based on viable counts and showed that numbers of surviving organisms decreased over time. Survival was inversely related to storage temperature and RH. Although the average counts decreased over time, a limited proportion of shells carried high numbers of SE at all storage conditions. 4. Penetration spots were observed earlier using an increased storage temperature due to increased growth rates of SE on the agar. After 20 d of storage a similar percentage (c. 44.7%) of eggshells became penetrated, irrespective of the storage conditions tested in this study. 5. The higher the Salmonella shell contamination at the end of storage, the higher the probability that the eggshell was penetrated. Salmonella shell counts exceeding 4 log cfu yielded more than a 90% probability of eggshell penetration occurring.

Animals↗

Bacillus sporothermodurans and other highly heat-resistant spore formers in milk.

A recent example of a micro-organism causing undesired growth in consumer milk is Bacillus sporothermodurans producing highly heat-resistant spores (HRS) which may survive ultra-high temperature (UHT) treatment or industrial sterilization. Molecular typing showed a heterogeneous group of farm isolates (non-HRS strains), but a clonal group of UHT isolates from diverse European countries and other continents (HRS-clone) suggesting a common source. During a survey of Belgian dairy farms for the presence of potentially highly heat-resistant spore formers, high numbers of these spores were detected in filter cloth, green crop and fodder samples. The strain collection showed a high taxonomic diversity with 18 potentially new species and with Bacillus licheniformis and Geobacillus pallidus as predominating species overall. Seventeen B. sporothermodurans isolates were identified, mainly originating from feed concentrate. Heat resistance studies showed the UHT resistance of B. sporothermodurans spores present in industrially contaminated UHT milk, but a lower heat resistance of laboratory-grown strains (HRS and non-HRS). Hydrogen peroxide, used as sanitizer in the dairy industry, was found to induce higher heat resistance of laboratory-grown B. sporothermodurans strains to a certain level. This indicates that sublethal stress conditions may affect the heat resistance. By transmission electron microscopy, structural differences at the spore level were found between HRS and non-HRS strains. The data indicate that the attainment of extreme heat resistance is rather multifactorial.

Aerobiosis↗

The effect of a commercial UV disinfection system on the bacterial load of shell eggs.

AIMS: To study the effect of UV irradiation on the bacterial load of shell eggs and of a roller conveyor belt. METHODS AND RESULTS: The natural bacterial load on the eggshell of clean eggs was significantly reduced by a standard UV treatment of 4.7 s; from 4.47 to 3.57 log CFU per eggshell. For very dirty eggs no significant reduction was observed. Eggs inoculated with Escherichia coli and Staphylococcus aureus (4.74 and 4.64 log CFU per eggshell respectively) passed the conveyor belt and were exposed to UV for 4.7 and 18.8 s. The reduction of both inoculated bacteria on the eggshell was comparable and significant for both exposure times (3 and 4 log CFU per eggshell). Escherichia coli was reduced but still detectable on the conveyor rollers. The internal bacterial contamination of eggs filled up with diluent containing E. coli or S. aureus was not influenced by UV irradiation. CONCLUSIONS: There is a significant lethal effect of UV irradiation on the bacterial contamination of clean eggshells and recent shell contamination, contamination of rollers can be controlled and the internal contamination of eggs is not reduced. SIGNIFICANCE AND IMPACT OF THE STUDY: The penetration of UV into organic material appears to be poor and UV disinfection can be used as an alternative for egg washing.

Animals↗

Widespread endoscopic mucosal resection of the esophagus with strategies for stricture prevention: a preclinical study.

BACKGROUND AND STUDY AIMS: Circumferential endoscopic mucosal resection of the esophagus is complicated by stricture formation. Prophylactic measures for avoiding such strictures have not been well studied. The aim of this preclinical study was to assess strategies for prevention of esophageal strictures in a porcine model following widespread endoscopic mucosal resection (EMR). METHODS: A total of 18 60-kg pigs were included in the study. The roles of strip width (group 1), prophylactic steroids (group 2), and prophylactic stents (group 3) in the prevention of post-mucosectomy strictures were studied. Six animals were included in each group. Esophageal mucosal resection was achieved using a novel widespread EMR technique previously described by our group. Animals in group 1 underwent partial (50% circumference) mucosal resection without prophylactic measures, while animals in the other two groups underwent circumferential mucosal resection. Animals in group 2 received 80 mg of triamcinolone injected directly into the exposed submucosal tissue (20 mg injection in four quadrants). Animals in group 3 received esophageal metal stents coated with small-intestine submucosa (SIS) that were deployed immediately post-resection. Animals were kept alive for 1 month. RESULTS: Partial and circumferential widespread EMRs were achieved in all animals. There were no procedural complications. Repeat endoscopy at 1 month showed no strictures in group 1. Only four animals were studied in group 2, owing to the high complication rate (periesophageal abscess in all animals) with one early death. Three of the surviving animals developed mild to tight strictures. In group 3, all animals developed tight strictures; however, there was early stent migration in four animals and premature stent removal in two animals because of persistent vomiting. CONCLUSIONS: Partial widespread EMR of the esophagus heals without stricture formation and does not require prophylactic intervention. The use of deep mural steroid injection following a circumferential resection does not appear to prevent strictures and may result in serious adverse events. Short-term use of esophageal stents is inadequate for stricture prevention. However, better results may be anticipated with longer term (at least 6 weeks) stent use.

Animals↗

Bacterial eggshell contamination in conventional cages, furnished cages and aviary housing systems for laying hens.

The influence of housing system on the initial bacterial contamination of the eggshell was studied. Two long-term experiments were performed. Bacterial eggshell contamination, as expressed by total count of aerobic and Gram-negative bacteria, was periodically analysed for eggs from a conventional cage, a furnished cage with nest boxes containing artificial turf or grids as nest-floor material and an aviary housing system. Results were log-transformed prior to statistical analyses. For both experiments no systematic differences were found between the conventional cage and furnished cage. The type of nest-floor material in the nest boxes of the furnished cages also did not systematically influence the bacterial contamination. A possible seasonal influence on contamination with a decrease in the winter period (up to > 0.5 log cfu/eggshell) of total count of aerobic and Gram-negative bacteria was observed in the first experiment. The contamination with total aerobic flora was higher (more than 1.0 log) on eggs from the aviary housing system compared to the conventional and the furnished cage systems. For Gram-negative bacteria this was not the case. During the entire period of both experiments, independent of housing system, shell contamination was not influenced by age of hens or period since placing the birds in the houses. For the total count of aerobic bacteria a restricted positive correlation (r2 = 0.66) was found between the concentration of total bacteria in the air of the poultry houses and initial shell contamination.

Air Microbiology↗

Eggshell characteristics and penetration by Salmonella enterica serovar Enteritidis through the production period of a layer flock.

1. Egg weight, shell thickness, number of pores, cuticle deposition and ability of Salmonella enterica serovar Enteritidis (SE) to penetrate the shell were determined for eggs from one layer flock through the entire production period. 2. Penetration was assessed by filling the eggs with a selective medium that allowed visualising Salmonella growth on the inside of the shell and membrane complex. After inoculation of each shell with on average 2.59 log cfu, the eggs were stored for up to 20 d at 20 degrees C and 60% relative humidity (RH). 3. On average 38.7% of the eggshells became penetrated. Mostly penetration occurred on d 3. Although it affected all shell characteristics studied, hen age did not significantly influence eggshell penetration. 4. No correlations were observed between any of the shell characteristics studied and the ability of SE to penetrate the shell. The growth of SE on the shell is of major importance because shell contamination at 20 d of storage and SE penetration were highly correlated.

Aging↗

Species identification of poultry egg products.

Species-specific primers for duck were deduced from the mitochondrial ATPase8 gene sequence. Species-specific PCR for turkeys and ducks showed no cross reaction with mixtures from chicken and guinea eggs and detection was possible to a concentration of 0.1% of homogenized duck egg and 5% of homogenized turkey egg when a PCR of 35 cycles was applied. A PCR of 30 cycles detected 10% of homogenized duck egg. The same sensitivities were obtained in dilutions of homogenized egg yolk; however, no PCR signals were obtained in egg white. Analysis of 13 industrial egg product samples showed the practical relevance of the species-specific PCR tests described herein.

Adenosine Triphosphatases↗

Routes for Campylobacter contamination of poultry meat: epidemiological study from hatchery to slaughterhouse.

From April 1998 to March 2000, 18 broiler flocks were followed from the hatchery to the slaughterhouse. Campylobacter was not found in the hatchery, 1-day-old chicks or in the rearing house before the arrival of the chicks. The infection of broiler flocks increased continuously during the rearing time, with a total of seven positive flocks at the end of rearing. Farms with Campylobacter-positive broilers were characterized by the circulation of Campylobacter in the environment (puddles, dung hill) and on the footwear of the farmer. The administration of antibiotics did not significantly reduce Campylobacter shedding. With the exception of one flock during rearing and a few flocks in the slaughterhouse with a mixed Campylobacter coli-Campylobacter jejuni infection, C. jejuni exclusively was found both during rearing and on the carcasses. A significant correlation exits between the contamination of the broilers during rearing and the carcasses after processing. No slaughterhouse was able to avoid contamination of carcasses when status-positive animals were delivered. Moreover, six negatively delivered flocks yielded positive carcasses, the result of a supplementary contamination, which occurred during transport and slaughtering.

Abattoirs↗

Salmonella on pig carcasses: positive pigs and cross contamination in the slaughterhouse.

AIMS: The purpose of this study was to investigate the prevalence of Salmonella in pigs at the moment of slaughter and in the slaughterhouse environment. METHODS AND RESULTS: In total, five different commercial slaughterhouses were sampled during eight slaughterhouse visits. Carcass swabs, colon content and mesenteric lymph nodes were taken to reflect the animal status and from the slaughterhouse environmental samples were taken. Salmonella was isolated from 37% of the carcass samples as a mean value. High variations were noticed between different slaughterhouses (between 0 and 70%) and sampling days in the same abattoir (between 3 and 52%). A correlation was found between the carcass contamination and the status of the delivered animals (P=0.01675). Cross contamination was estimated to account for 29% of the positive carcasses. The slaughterhouse environment was highly contaminated; before starting the slaughtering activities 25% of the samples were positive on average. The most prevalent serotypes isolated at the slaughterhouse environment and from the colon content were S. Typhimurium, S. Livingstone and S. Derby. On carcasses S. Typhimurium was predominately isolated (71%). The biggest variability of serotypes was found in the mesenteric lymph nodes. Serologically 56.3% of the pigs were found positive for Salmonella using a cut-off level of the optical density percentage higher than 10 (O.D.% > or = 10). While on individual pig level the correlation between the bacteriological and serological data was poor, because of recent Salmonella infections, a better correlation was found at the herd level on the moment of slaughtering. CONCLUSION: A high degree of carcass contamination is noticed after slaughtering. This contamination resulted from the delivery of Salmonella-positive pigs and cross-contamination from the slaughterhouse environment. SIGNIFICANCE AND IMPACT OF THE STUDY: In pigs, Salmonella carriage is high, but it is obvious that slaughterhouse hygiene is a determinative factor for managing carcass contamination.

Abattoirs↗

Routes for salmonella contamination of poultry meat: epidemiological study from hatchery to slaughterhouse.

Data were collected on the prevalence of salmonella at different stages during the life cycle of 18 broiler flocks on different farms as well as during slaughter in different poultry slaughterhouses. For the isolation of salmonella, the highest sensitivity (93.9%) was obtained by enrichment in the semi-solid agar Diasalm. The 'overshoe method' utilizing several pairs of overshoes provided the highest sensitivity for determining the salmonella status of the broilers during rearing. A clear decrease of the relative importance of the first production stages was demonstrated for the salmonella contamination of the end product, whereas horizontal transmission of salmonella to broilers during rearing and to broiler carcasses in the slaughterhouse was shown to be the main determinative factor. Ten of the 18 flocks received a salmonella positive status with the highest shedding occurring during the first 2 weeks of rearing. The shedding of the animals was significantly negatively influenced by the use of subtherapeutic or therapeutic doses of antibiotics. The intake of portable material in the broiler house was identified as the most important risk factor for horizontal transmission. Significant associations were found between the contamination level of a flock and hygiene of the broiler house, feed and water in the broiler house and both animal and non-animal material sampled in the environment. No correlation was found between contamination during the rearing period and contamination found after slaughtering. The presence of faecal material in the transport crates and predominantly the identity of the slaughterhouse seemed to be the determining factors for carcass quality. Improved hygiene management during transport of broilers and in some slaughterhouses could significantly reduce the risk of salmonella contamination of poultry meat.

Animal Husbandry↗

Polymerase chain reaction identification of Bacillus sporothermodurans from dairy sources.

AIMS: A new polymerase chain reaction (PCR) method for the identification of Bacillus sporothermodurans strains from sterilized or ultrahigh temperature-treated milk and milk products and from other non-milk sources and environments, including the dairy farm. METHODS AND RESULTS: Two strains from raw milk and feed concentrate could be allocated to B. sporothermodurans based on 16S rDNA sequencing and DNA-DNA hybridization results. Two specific PCR primers were derived from the 16S rRNA gene of B. sporothermodurans. CONCLUSIONS: The PCR identification method was validated using a collection of B. sporothermodurans strains from different sources and on a large collection of dairy and non-dairy Bacillus spp. and other relevant taxa. SIGNIFICANCE AND IMPACT OF THE STUDY: This PCR method was used as a screening method for strains with very heat-resistant endospores, isolated at the dairy farm level after heat treatment for 30 min at 100 degrees C. Seventeen strains isolated at the dairy farm were identified as B. sporothermodurans. They originated mainly from feed concentrate and also from soy, pulp and silage. The PCR identification method described here can, therefore, contribute to a better understanding of the route by which B. sporothermodurans contaminates raw and/or heat-treated milk.

Animal Feed↗

Evaluation of the insulin resistance syndrome in 5- to 10-year-old overweight/obese African-American children.

OBJECTIVE: To characterize the insulin sensitivity of overweight and obese 5- to 10-year-old (Tanner stage 1-3) African-American children screened for participation in a diabetes prevention study and to identify the association of insulin sensitivity with obesity, hyperlipidemia, and hypertension. RESEARCH DESIGN AND METHODS: Measures of insulin resistance (homeostasis model assessment) and insulin sensitivity (Matsuda and DeFronzo's whole-body insulin sensitivity) were calculated from a 2-h oral glucose tolerance test in 137 African-American children recruited into a diabetes prevention study. Measures of lipids (LDL, HDL, total cholesterol, and triglycerides), blood pressure, and body composition were obtained for a subset of the children. RESULTS: In response to a glucose challenge, girls and older and heavier children produced significantly more insulin. As BMI increased, there was a statistically significant decrease in insulin sensitivity, particularly in girls. Insulin sensitivity was inversely correlated with increases in blood pressure, triglycerides, subcutaneous fat, the percentage of total body fat, and Tanner stage, but it was not correlated with LDL and HDL. CONCLUSIONS: Reduced insulin sensitivity and the cluster of risk factors known as the insulin resistance syndrome (IRS) are already apparent in these overweight African-American children. Young African-American girls, in particular, already show evidence of hyperinsulinemia in response to a glucose load, suggesting that the early stages of metabolic decompensation that lead to type 2 diabetes are already occurring. Monitoring of those risk factors known to be part of IRS should become part of routine medical care for overweight or obese African-American children.

Adipose Tissue↗

PCR-TGGE: a method for fingerprinting the microbial flora in the small intestine of pigs.

The bacterial flora of the small intestine is still not fully understood. This is also due to the fact that a lot of these gut inhabitants aren't cultivable. PCR-TGGE (Temperature Gradient Gel Electrophoresis) is a molecular tool by which the 16S ribosomal DNA of a bacterial community can be studied for its diversity. Based on sequence differences in the variable region V6-V8, 16S ribosomal DNA fragments are separated during a TGGE run. This study shows that PCR-TGGE is a valuable tool as it gives additional information about the flora composition that could not be found with classical bacteriological techniques.

Animal Feed↗

Prevalence of Salmonella, Campylobacter and VTEC on pig farms.

Four different pig farms were sampled for the prevalence of Salmonella, Campylobacter and verotoxigenic E. coli (VTEC). Pigs of different age groups and pigs of the same age were tested by taking rectal swabs. The farm environment was tested by examining overshoes, and the feed and drinking water in the pens. 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 positive 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. The presence of Campylobacter was tested in 150 rectal swabs, 51 of these, spread over the four farms, turned out to be positive. At all four pig farms Campylobacter was isolated from the environment as well. 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 wich causes the endema disease in young pigs, four strains harboured the hlyA gene and 19 the eaeA gene.

Adhesins, Bacterial↗