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Dissertation
Aanrijking van omega-3 en omega-6 vetzuren uit visolie
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Year: 1990

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Hygiëne in verrijkte kooien versus niet-kooisystemen voor leghennen
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Year: 2008 Publisher: [Anderlecht Pluimvee]

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Dissertation
Occurrence and control of Listeria monocytogenes in the beef and pork supply chain
Authors: --- --- --- ---
Year: 2022 Publisher: Leuven KU Leuven. Faculty of Bioscience Engineering

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Despite the efforts already made by the sector, slaughterhouses and cutting plants are still confronted with the foodborne pathogen Listeria monocytogenes in the production environment and on carcasses and meat cuts. Its occurrence may result in meat contamination, leading to economic losses and a potential risk for public health, as evidenced by recent outbreaks. Therefore, there is a need for scientifically based knowledge to provide these companies with strategies to control better this foodborne pathogen, whose introduction and variable contamination routes remain challenging to characterize.This work aims at filling knowledge gaps on the subject. To this end, we first mapped the L. monocytogenes occurrence on carcasses in 7% of the Belgian slaughterhouses. The pathogen was repeatedly recovered from beef (47%) and pig (20%) carcasses just before cooling. Microbial contamination of carcasses differed substantially according to the location on the carcass. Ventral and anterior sites were more contaminated on pig carcasses, probably associated with inadequate decontamination in the unclean zone or potential post-contamination during evisceration. As for cattle carcasses, contaminated sites such as hind leg, flank, brisket, and foreleg suggest pathogen transfer during (manual) dehiding practices. Moreover, in one of the slaughterhouses, the beef carcasses were contaminated entirely with a specific strain (belonging to CC9), suggesting a persistent source of contamination along the slaughter line.We further investigated contamination routes in beef slaughterhouses by sampling tracked animals/carcasses and the environment. The hides were contaminated with L. monocytogenes in 93% of the incoming animals, signifying high contamination pressure levels entering the slaughterhouses. Sporadic pathogen transfer to carcasses during dehiding was confirmed by identifying the same strains from hides, carcasses, and utensils. Still, persistence of one strain (belonging to CC9) in a carcass splitter showed a more significant problem, causing constantly repeated contamination of carcasses over more than one year.Furthermore, we investigated the occurrence and genetic diversity of L. monocytogenes in pork cutting plants by thoroughly sampling the processing environment to identify harborage sites. Non-food contact surfaces (zone 3) were contaminated more frequently at typical harborage sites. Although less frequently contaminated, food contact surfaces (zone 1) were also positive for detecting this pathogen, even after cleaning and disinfection. The isolated strains showed low genetic heterogeneity. Some identical pulsotypes belonging to typical clonal complexes (CC8, CC9, CC31, and CC121) were predominant and widespread, suggesting persistence rather than occasional introduction and repeated contamination.Lastly, we carried out challenge test studies according to the EURL Lm Technical Guidance Document for conducting shelf-life studies on L. monocytogenes in ready-to-eat foods. The growth potential was determined in pork chops and minced pork under packaging and storage conditions that mirror retail and home conditions. Pork chops did not support the growth of the pathogen (<0.5 log CFU/g) under the applied conditions (air and MAP). Substantial growth of L. monocytogenes (>0.5 log CFU/g) was obtained in minced pork for air-packaging and MAP. However, significant intra- and inter-batch variability was observed, raising questions about how the growth potential is calculated. Additionally, the maximum growth rate in minced pork at a constant temperature of 7°C was determined in air and MAP for simulation studies. Determining the growth potential in minced pork using these models, as well as the Combase Growth model and DMRI dynamic safety model, gave overestimations, especially for MAP. In our view, the models used do not sufficiently consider the raw pig matrix dynamics, which may have an inhibitory effect on the growth of L. monocytogenes. On the other hand, the experimentally high variability impedes the utility of purely deterministic predictive microbiology models in these raw pork products and probabilistic approaches are essential to reflect reality.In conclusion, this research has provided insights on contamination sources and routes, critical points, persistence, and growth of this pathogen to help the sector develop concrete action plans and better control the problem.

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Dissertation
Occurrence and characterisation of residual contamination and biofilms in food processing environments and poultry drinking water systems
Authors: --- --- --- ---
Year: 2018 Publisher: Leuven KU Leuven. Faculty of Bioscience Engineering

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Despite a regular cleaning and disinfection (C&D), residual bacterial contamination and biofilms still occur in food companies and primary animal production environments. In case pathogenic and spoilage organisms remain, these could lead to early food spoilage, foodborne illness and pose a threat for animal and human health. Further knowledge concerning the occurrence, composition and characteristics is needed to have a better estimate on the impact of this residual contamination. Therefore, this research aimed at gaining insights in the presence and characteristics of residual bacterial contamination and biofilms in different food sectors and in primary animal production, more specifically the drinking water system (DWS) in broiler houses.In the first part of the research (chapters 3 and 4), biofilms and residual bacterial contamination in eight food companies of different sectors were mapped and investigated. To do so, there was a need for a suitable sampling method that allowed detection and/or quantification of microorganisms and biofilm matrix components. Two surface sampling methods were tested, the sponge stick method and the scraper-flocked swab method, whereby the latter method was evaluated as the most suitable. Sampling with this method provided results of the bacterial load and chemical composition of surface contamination after C&D. Bacterial enumerations were on average 3.62 &plusmn; 1.20 log CFU/100 cm² but reached up to 7.23 log CFU/100 cm². Respectively 20%, 15% and 8% of the surfaces investigated for biofilm matrix components were contaminated with low quantities of proteins, carbohydrates and uronic acids. On 17% of the investigated surfaces, both microorganisms and at least one of the chemical compounds were found, which is an indication for the presence of biofilm. The presence and the degree of residual contamination is highly variable by food sector, food company, sampling point and even sampling time. Genera that were most abundant in the residual bacterial contamination on food contact surfaces after C&D in the different food companies were Pseudomonas, Microbacterium and Stenotrophomonas, however 60% of the identified genera were company-specific. Of all the evaluated isolates, 88% had some kind of spoilage potential, with the ability to break down lipids as the most prevalent property. However it remains difficult to estimate the possible impact of these microorganisms on food safety and spoilage since it is not known whether they will: be transferred from the surface to the food products, survive and grow in the food products and consequently cause spoilage.In the second part of the research (chapters 5 and 6), the focus was on residual contamination and biofilm formation on the inside of the DWS in broiler houses after disinfection. Water quality in the DWS plays an important role in the general health and performance of broiler chickens since pathogens might be present. Conditions in the DWS of broilers are ideal for microbial biofilm formation. The presence of this contamination on the inside of the DWS was assessed in terms of bacterial load and chemical composition. Average bacterial counts of 6.03 &plusmn; 1.53 log CFU/20cm² were observed, ranging up to 9.00 log CFU/20cm² at some points. Proteins, carbohydrates and uronic acids were again found in low quantities in 58%, 14% and 5% of the samples, respectively. On 63% of the investigated surfaces, the presence of biofilm was suspected since microorganisms were detected in combination with at least one of the analysed chemical components. The most identified dominant species in the DWS were Stenotrophomonas maltophilia, Pseudomonas geniculata and Pseudomonas aeruginosa, which are opportunistic human pathogens. However at species level, most of the identified microorganisms were farm-specific. Almost all the isolates belonging to the three most abundant species were strong biofilm producers. Overall, 92% of all tested microorganisms were able to form biofilm in 96-well microtiter plates. Finally, the hypothesis that commensal bacteria in biofilms on surfaces could prevent attachment of pathogens such as Salmonella spp. was investigated. Since there is an increasing problem with Salmonella Java contamination on broiler chickens in Belgium, this pathogen was used to investigate its interaction with the commensal Pseudomonas putida, which is part of the natural microbiota in the DWS. Therefore, a new model that simulates biofilm formation on the inside of the DWS was developed and validated. In this model, Salmonella Java was evaluated as a strong biofilm former. However, when applied in the presence of Pseudomonas putida, biofilm formation by Salmonella Java was reduced due to competitive interactions indicating the potential of Pseudomonas putida as a biocontrol agent.In conclusion, this research provided interesting new information for food companies to be used in their fight against unwanted contamination and for the development of more efficient C&D procedures. Also in primary animal production, new insights concerning the presence and composition of DWS contamination and its role in the prevention of pathogens were obtained.

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