Inactivation of classical swine fever virus in porcine casing preserved in salt.

Pig intestines used for the production of natural sausage casings may carry classical swine fever (CSF) virus. Feeding pigs with human food waste that contains pig casings may then spread the virus to CSF-free animals. Casings derived from a pig experimentally infected with CSF by dosing with 10(6) tissue culture infectious doses (TCID50) of the highly virulent CSF virus strain "Koslov", were treated with phosphate supplemented or citrate supplemented NaCl, instead of with NaCl alone, which is the standard preservation treatment for casings. Treated casings were stored for 30 days at either 4 degrees C or 20 degrees C. After storage the casings were fed to 16 susceptible pigs. CSF infection was confirmed in the four animals that had been fed casings treated with citrate supplemented salt and stored at 4 degrees C. All other animals remained healthy. It is therefore possible to avoid the inadvertent spread of CSF virus via porcine sausage casings by treating casings with phosphate supplemented salt and storing them for 30 days at temperatures over 4 degrees C.

Quantitative histological analysis of bovine small intestines before and after processing into natural sausage casings.

A histological study was undertaken to determine the efficiency in the removal of the mucosa and Peyer's patches by standard processing of bovine intestines into natural sausage casings. The second objective was to calculate the quantity of lymphoid and nervous tissue per consumable sausage. For the histological analysis, intestinal samples were collected from 80 beef cattle during the slaughter process. Fresh and cleaned intestines were compared in analyzing the thickness of the intestinal wall, weight reduction during cleaning, removal of the mucosal layer, and the presence of lymphoid and neural tissue after cleaning. The obtained data indicate a weight reduction of about 50% during standard cleaning procedures, as 90% of the mucosa and 48% of the lymphoid tissue are removed. Based on the quantitative histological image analysis, it was calculated that 1 m of cleaned casings, weighing on average 64 g, contains about 2.8 g of mucosa, 0.3 g of lymphoid tissue, and 0.1 g of neural tissue. Assuming, in a worst-case scenario, that the sausage casing is ingested when consuming 200 g of sausage at one meal, this consumption includes 0.09 g of lymphoid tissue and 0.02 g of neural tissue as part of the sausage casing. These data can be included in a risk assessment on the potentialexposure of consumers to bovine spongiform encephalopathy infectivity after eating sausages in beef casings.

Removal of foot-and-mouth disease virus infectivity in salted natural casings by minor adaptation of standardized industrial procedures.

Intestines are used for the production of natural casings as edible sausage containers. Derived from animals (pigs and sheep) experimentally infected with FMDV (initial dosage 10(7.3) PFU/ml, strain O(1Kaufbeuren)), these natural casings were treated with sodium chloride or a phosphate salts/sodium chloride mixture and the residual FMDV titres measured. After storage at about 20 degrees C, no remaining infectivity was found after either treatment, whereas casings stored at 4 degrees C still contained infectivity. Storage of salted casings at about 20 degrees C for 30 days is already part of the Standard Operating Procedures (included in HACCP) of the international casing industry and can therefore be considered as a protective measure for the international trade in natural casings.

Suggestions for the construction, analysis and use of descriptive epidemiological models for the modernization of meat inspection.

There is consensus that scientifically validated, quantitative assessments of actual public health risks are a prerequisite for any sound modernization of current meat inspection procedures. This article outlines how such analyses could be conducted. Approaches that rely heavily upon extrapolations from theoretical dose-effect relationships are inadequate for the assessment of microbiological health risks associated with the production and consumption of meat. The use of highly structured and very elaborate descriptive epidemiological models covering the entire period from stable to table can be considered a promising solution. Health risks can be quantified by means of incidence rates and the influence of risk factors by means of odds ratios and (population) attributable fractions. A great advantage is that when it is not possible to quantify risks exactly, the descriptive models are detailed enough to be used in a hazard analysis critical control point (HACCP)-like approach and for writing validated codes of good manufacturing practice (GMP). There are, however, several conditions which have to be met before risk assessment can become the foundation of safety assurances for meat, such as active legislative support and the setting up of monitoring systems for zoonoses and other health hazards in animals and humans.

Identification and quantification of risk factors regarding Salmonella spp. on pork carcasses.

The main elements of a descriptive epidemiological model for Salmonella spp. in Dutch pig slaughterlines, and the subsequent quantification of risk factors regarding the contamination of carcasses, are described. There is a strong correlation between the number of live animals that carry Salmonella spp. in their faeces and the number of contaminated carcasses at the end of the slaughterline. Live animals that carry Salmonella spp. are 3-4 times more likely to end up as a positive carcass than Salmonella-free animals. Currently, about 70% of all carcass contamination results from the animals themselves being carriers, and 30% because other animals were carriers (i.e. cross contamination). Furthermore, it is estimated that in general between 5-30% of the carcasses produced may contain Salmonella spp. With respect to carcass contamination with Enterobacteriaceae and Salmonella spp., inadequately cleaned polishing machines (odds ratio, OR, 6) and 'inapt procedures during evisceration' (OR 11), i.e. faulty evisceration and hygiene practices, are the most important risk factors. An estimated 5-15% of all carcass contamination with Salmonella spp. occurs during polishing after singeing. The remainder is the result of current evisceration practices (55-90%) and, to a lesser extent, further processing (5-35%), i.e dressing, splitting and meat inspection. Less likely Salmonella spp. already present on the skin of the live animals survive scalding and singeing. However, because pigs are the only important source for the Salmonella contamination of the line and the carcasses produced, it can also be concluded that if Salmonella-free pigs were produced, consumers could be provided with virtually Salmonella-free pork. As long as Salmonella-positive animals enter abattoirs, there will always be transmission of Salmonella spp. to consumers, even if the process is carried out according to stringent codes of good manufacturing practices (GMP). EU regulations should, therefore, allow for the decontamination of caracasses with a safe substance, e.g. lactic acid, on the condition that the slaughterhouse strictly adhers to GMP principles.

Salmonella spp. on pork at cutting plants and at the retail level and the influence of particular risk factors.

This article describes the contamination of pork with Salmonella spp. in cutting plants and butchers' shops in The Netherlands and quantifies the influence of several risk factors. When contaminated carcasses are being processed, the main risk factors regarding cross contamination are inapt cleaning and disinfection (OR 12.8), manipulation of contaminated materials as such (OR 4.7) and (re)contaminated surfaces (OR 4.4). However, in the current situation, where contaminated carcasses are constantly being brought into cutting lines, interim cleaning and disinfection of surfaces and utensils during breaks and at the end of the working day will most likely prevent not more than about 10% of all cross contamination that takes place during a working day. Thus, as long as contaminated carcasses are being processed, about 90% of the cross contamination that occurs in cutting plants is practically unavoidable. It can therefore also be concluded that under these circumstances the implementation of codes of good manufacturing practices (GMP) and Hazard Analysis Critical Control Point (HACCP)-inspired production methods will only be marginally effective in the control of Salmonella spp. cross contamination in cutting lines. The same is more or less true for the processing of contaminated cuts or carcasses by butchers in shops and supermarkets. Furthermore, in contrast to the situation in cutting plants, it may be that up to 10% of butcher's shops or kitchens of restaurants become colonized for several weeks or months with their own endemic 'house flora' of Salmonella spp., which are originally introduced via the purchased contaminated products of animal origin. Though there are no hard data to substantiate this, it can be suspected that these shops and restaurants represent the more badly managed, i.e. poorly cleaned and disinfected, enterprises. However, several analytical limitations hinder an exact determination of the prevalence of Salmonella spp. contaminated pork and an exact quantification the influence of risk factors. The diagnostic value (i.e. the sensitivity, specificity, precision and predictive value) of the combination of swabbing of carcasses and cuts and the usually employed culturing methods, in particular, is largely unknown, and there are indications that it may be seriously questioned. Without a more thorough knowledge about the diagnostic value of current and future methods of sampling and identification, it is impossible to provide for more accurate estimations of the prevalence of Salmonella positive carcasses and cuts. Based on the research data, the incidence of contaminated cuts and retail-ready pork can not be estimated more precise than as somewhere between 5-40%. When compensating for the discussed methodological flaws, it must be assumed that currently the true prevalence of contaminated primal cuts and retail-ready pork in butchers' shops is about 25-30%, and that of minced pork and pork sausages about 50-55%. Lastly it is concluded that if carcasses were Salmonella-free, consumers could in principle be provided with virtually Salmonella-free pork. It is therefore recommended that the EU allows for a decontamination step in slaughterhouses with a substance that is generally recognized as safe, provided that the producers strictly adhere to GMP-principles.

Impact on human health of Salmonella spp. on pork in The Netherlands and the anticipated effects of some currently proposed control strategies.

The impact on human health of Salmonella spp. on pork in The Netherlands is described. Subsequently, the effects of some currently proposed control strategies in the Dutch pork production chain are evaluated and quantified with the aid of a simple mathematical model. The estimated average incidence of cases of salmonellosis in the Netherlands is about 450 cases per 100,000 person years at risk (pyar). Some special risk groups for which the risks could be quantified are (1) persons with underlying diseases, such as neoplasms or diabetes mellitus (1200 cases/100,000 pyar); (2) persons with achlorhydria or who excessively use antacids (1100 cases/100,000 pyar); (3) persons who have recently been treated with antibiotics that disturb the normal gut flora (1700 cases/100,000 pyar); (4) nurses (900 cases/100,000 pyar); (5) caterers (900 cases/100,000 pyar); (6) slaughterline personnel (1800 cases/100,000 pyar). Furthermore, it is estimated that 15% (5-25%) of all cases of salmonellosis in The Netherlands are associated with the consumption of pork. Currently, proposed control measures regarding Salmonella in pigs and on pork in The Netherlands are codes of good manufacturing practices (GMP) that, in fact, formalize recommendations that can be found in many handbooks about pig breeding and pig slaughtering. When evaluated by a mathematical model constructed for this purpose, the proposed GMP codes from farm to cutting/retail could, at best, reduce the current levels of Salmonella-positive pigs and pork by 50-60%. If pigs were bred according to the rather costly specific pathogen-free concept (SPF), the prevalence of contaminated carcasses and pork could in total be reduced by 95% or more. However, implementing GMP codes from the transport phase up to the cutting/retail phase coupled with a decontamination step at the end of the slaughterline would be just as effective as GMP in combination with breeding using the SPF-concept. It is therefore concluded that the most efficient and cost-effective way of reducing the 'Salmonella problem' entailed by the consumption of pork would be to decontaminate carcasses under the precondition that the entire production chain strictly adheres to GMP principles. Therefore, the EU should also allow for more possibilities regarding the decontamination of carcasses than is currently the case. It is also concluded that current EU regulations relying on hazard analysis of critical control points (HACCP)-inspired production in cutting plants will not be effective in reducing the prevalence of Salmonella spp. on pork. This is mainly because (1) there is currently an almost steady stream of Salmonella-positive carcasses that enter the cutting process; (2) when contaminated carcasses are being processed, further cross-contamination during working hours is unavoidable; (3) no steps in the cutting process are intentionally designed to effectively reduce the risks or consequences of cross contamination of cuts and retail-ready products.

Identification and quantification of risk factors in animal management and transport regarding Salmonella spp. in pigs.

This article discusses the main elements of a descriptive epidemiological model for Salmonella spp. in the pre-harvest stages of pork production, and the subsequent quantification of risk factors. About two thirds of all Dutch pig farms are more or less permanently infected. At infected finishing farms, the current probability that Salmonella-free pigs will become infected is about 85%. In the case that a certain pen is infected, the current probability that further pen-to-pen transmission will occur is about 90% and that (human) vectors will also spread the infection 60%. Between 5-30% of the animals may still excrete Salmonella spp. at the end of the finishing period, and this percentage can double during transport and lairage. In infected pigs at slaughter weight, the Salmonella spp. are foremost located in the digestive tract, its contents and the closely associated lymph nodes. Under the current circumstances, the lack of farm hygiene (odds ratio (OR) 39.7), (re)contaminated feed (OR 1.6), the use of broad spectrum antibiotics (OR 5.6), a positive Salmonella-status of animals before transport (roughly estimated OR 4.0), the lack of transport hygiene (roughly estimated OR 1.1) and transport stress (OR 1.9) are the most important risk factors regarding infections with Salmonella spp. Currently the role of on-farm contamination cycles with endemic ('house flora') Salmonella spp. is so important, that the role of other factors is difficult to ascertain. It is also concluded that the farm-phase forms the core of all current problems, and that better prevention and control can be achieved by, (i) very strict and consistent farm hygiene in combination with promotion of the colonization resistance of animals kept together with a prudent use of broad spectrum antibiotics; (ii) simultaneous execution of control programmes at breeding farms, multiplying farms and finishing farms; (iii) separate transport, lairage and slaughter of the animals thus produced.

An outline of a risk assessment-based system of meat safety assurance and its future prospects.

Discussed are the outlines of a risk assessment-based system of meat safety assurance to replace the current meat inspection. An example of a system that uses the Hazard Analysis of Critical Control Points (HACCP)-principles in the entire production chain from stable to table is also given. Continuous evaluation of risks is the main driving force of the new system. Only then the system has the means to remain flexible and provide for the data necessary to convince trade partners that the products they buy are safe. A monitoring system that keeps track of the important health hazards in the entire chain from stable to table is therefore necessary. This includes monitoring of cases of disease in the human population caused by the hazardous agents of concern. Coordination of the monitoring and control and processing of the information is done by an independent body. Furthermore, the system demands a production from stable to table that is based on the ideas of Integrated Quality Control (IQC), HACCP, and certification of production processes and quality control procedures. Clear legislation provides for criteria about acceptable or unacceptable health risks for the consumer and determines at what moments which risks should be controlled by the producers. Simultaneously, the legislation has to be flexible enough to be able to adapt quickly to any changes in risks, or in the way risks should be controlled. In the new system current meat inspection can easily be carried out by employees of the slaughter houses and is no longer a direct responsibility of the authorities. The authorities only demand certain safety levels and verify whether producers stick to these. Producers remain fully responsible for the safety and quality of their products, and fully liable in case of any damage to the consumers' health. However, it is to be expected that some EU Member-States miss the organizational and agricultural basis for a successful application of the new system. Consequences are that two parallel flows of meat and meat products may come to existence. One flow will exist of meat produced according to the new, and with respect to safety assurance, superior system. The other flow will consist of meat produced and inspected in the traditional way. However, this meat will contain all the flaws that required a revision of the system in the first place. This, and the fact that EU policy ordains that part-taking in an alternative meat inspection system should be voluntary, may very well result in a very slow start-up of the new system. One of the easiest solutions, however, would be to implement a decontamination step in the slaughtering process, provided that this is accompanied by strict codes of hygienic practices and good manufacturing practices. Not only would this lead to safer meat, but also result in the two separate flows of meat becoming one flow again as well as an easier to organize livestock production according to HACCP-principles.

Investigations with enzyme-linked immunosorbent assays for Trichinella spiralis and Toxoplasma gondii in the Dutch 'Integrated Quality Control for finishing pigs' research project.

In the framework of the Dutch field trial 'Integrated Quality Control (IQC) for finishing pigs' ELISA (screening) techniques were used to detect animals seropositive for Toxoplasma gondii and/or Trichinella spiralis. The aim was to determine whether farms which consistently delivered seropositive pigs could be detected and monitored (defined as 'problem farms'). The investigation involved 120 farms and three slaughterhouses, and a total of 23,348 serum samples were examined. In addition, all pigs were also screened for the presence of Trichinella spiralis with the digestion method (pooled samples). The prevalence of seropositivity for Trichinella spiralis and Toxoplasma gondii was 0.3% and 2.1% respectively. Parasitological examinations concerning T. spiralis were negative. Considering the characteristics of the used methodology, the conclusion was drawn that there were no parasitological or serological indications for T. spiralis infections, and that with respect to T. gondii the infection rate seemed to be equally low for all farms involved. In addition, a longitudinal pilot study during a whole finishing period was undertaken at two finishing farms. Animals seropositive for Toxoplasma gondii were found from the earliest days of the finishing period. Housing and management may (still) play an important role in the prevention of contact with this parasite.

Human health hazards associated with the administration of antimicrobials to slaughter animals. Part II. An assessment of the risks of resistant bacteria in pigs and pork.

Risks for the consumer regarding the acquisition of resistant bacteria and/or resistance genes via the consumption of pork are discussed. In general, Salmonella spp. and Escherichia coli that originate from animals do not easily transfer their resistance genes to the resident intestinal flora of humans. The prevalence of resistant E. coli in humans seems more associated with being a vegetarian (odds ratio (OR) 1.89) than with the consumption of meat and meat products. Other risk factors are treatment with antimicrobials (OR 2-5), becoming hospitalized (OR 5.93), or working in a health setting (OR 4.38). In the Netherlands, annually an estimated 45,000 people (0-150,000) become a carrier of resistant E. coli and/or resistance genes that ori ginate from pigs, while an estimated 345,000 persons (175,000-600,000) become a carrier of resistant E. coli and/or resistance genes that originate from hospitals, e.g. other patients. Any problems with resistant Salmonella spp. that stem from pigs are, in fact, an integral part of the total problem of food-borne salmonellosis. Sometimes there are outbreaks of a specific multi-resistant clone of S. typhimurium that causes problems in both farm animals and humans. The probability that in the next 30 years there is no or maximally one outbreak of a specific clone that originates from pig herds is estimated at about 75%. Antimicrobials used as a growth promoter can have a measurable influence on the prevalence of resistant bacteria. The likely chain of events regarding avoparcin and the selection and dissemination of resistance against vancomycin in the enterococci gives the impression that the impact of the use of antimicrobials in animals on the prevalence of resistance in humans is largely determined by whether resistance genes are, or become, located on a self-transferable transposon. Furthermore, consumer health risks of antimicrobials used in slaughter pigs are mainly determined by the selection and dissemination of bacterial resistance and much less by the toxicological properties of any residues in pork. It is also concluded that most of the problems with resistant bacteria in humans are associated with the medical use of antimicrobials, and that the impact of particularly the veterinary use of antimicrobials is limited. However, the impact of antimicrobials used as a feed additive appears to be much greater than that of antimicrobials used for strictly veterinary purposes. The use of antimicrobials as a feed additive should therefore be seriously reconsidered.

Human health hazards associated with the administration of antimicrobials to slaughter animals. Part I. An assessment of the risks of residues of tetracyclines in pork.

This article describes the assessment of consumer risks of residues of tetracyclines in slaughter pigs in the Netherlands. The assessed risks were toxic and allergic reactions, and the disturbance of the consumers' intestinal flora. Toxic and allergic reactions in humans and animals have only been observed at therapeutic doses, affecting between an estimated 1 in 5,000 and one 1 in 140,000 individuals exposed. Residues of tetracyclines in pigs are closely associated with treatment with injectable formulations. Established Maximum Residue Levels (MRLs) do not reflect actual consumer risks in case a limit is violated incidentally. For example, when the established MRLs for tetracyclines in meat are exceeded with a factor 400, 40,000, and 200,000, respectively, the actual risk of an adverse drug reaction for the consumer following a single consumption of this meat is maximally 1 in 3 million, 1 in 300,000, and 1 in 8,000, respectively. At the current estimated low levels of incidental exposure via pork, the annual risk of negative health effects for a random consumer is estimated at maximally 1 in 33 million. The annual risk that a temporary disturbance of the intestinal flora may also result in a facilitated infection with certain enteropathogens, such as Salmonella spp., is estimated at 1 in 45 million. It is concluded that the current microbiological risks of pork are greater than the risks of residues of tetracyclines as such, and that the control of the microbiological risks of pork should therefore be given first priority.

Efficacy of current EC meat inspection procedures and some proposed revisions with respect to microbiological safety: a critical review.

This paper presents an evaluation of the current EC meat inspection procedures, and some of their proposed revisions, in relation to their efficacy in assuring the microbiological safety and quality of meat, and the difficulties for health authorities and industry in providing such an assurance. It is concluded that neither the current nor the proposed revisions of ante and post mortem meat inspection procedures alone are sufficient, and that only integrated approaches, applied to each step of animal and meat production, will lead to better quality meat. Furthermore, for the design of a really effective and flexible long-term system of safety and quality assurance it is necessary to undertake a formal quantitative assessment of risk.

[The Hazard Analysis Critical Control Point approach (HACCP) in meat production]. / De Hazard Analysis Critical Control Point (HACCP)-benadering bij de produktie van vlees.

The Hazard Analysis Critical Control Point (HACCP) approach is a method that could transform the current system of safety and quality assurance of meat into a really effective and flexible integrated control system. This article discusses the origin and the basic principles of the HACCP approach. It also discusses why the implementation of the approach is not as widespread as might be expected. It is concluded that a future implementation of the approach in the entire chain of meat production, i.e. from conception to consumption, is possible. Prerequisites are, however, that scientifically validated risk analyses become available, that future legislation forms a framework that actively supports the approach, and that all parties involved in meat production not only become convinced of the advantages, but also are trained to implement the HACCP approach with insight.

Microbial risks associated with exposure to pathogens in contaminated urban flood water.

Urban flood incidents induced by heavy rainfall in many cases entail flooding of combined sewer systems. These flood waters are likely to be contaminated and may pose potential health risks to citizens exposed to pathogens in these waters. The purpose of this study was to evaluate the microbial risk associated with sewer flooding incidents. Concentrations of Escherichia coli, intestinal enterococci and Campylobacter were measured in samples from 3 sewer flooding incidents. The results indicate faecal contamination: faecal indicator organism concentrations were similar to those found in crude sewage under high-flow conditions and Campylobacter was detected in all samples. Due to infrequent occurrence of such incidents only a small number of samples could be collected; additional data were collected from controlled flooding experiments and analyses of samples from combined sewers. The results were used for a screening-level quantitative microbial risk assessment (QMRA). Calculated annual risks values vary from 5 x 10(-6) for Cryptosporidium assuming a low exposure scenario to 0.03 for Giardia assuming a high exposure scenario. The results of this screening-level risk assessment justify further research and data collection to allow more reliable quantitative assessment of health risks related to contaminated urban flood waters.

The price of the precautionary principle: cost-effectiveness of BSE intervention strategies in The Netherlands.

Since 1996, bovine spongiform encephalopathy (BSE) in cattle has been linked to a new variant of Creutzfeldt-Jakob disease (vCJD), a fatal brain disease in man. This paper assessed the cost-effectiveness of BSE control strategies instituted by the European Commission. In a Monte Carlo simulation model, a non-intervention baseline scenario was compared to three intervention strategies: removal of specified risk materials from slaughter animals, post-mortem testing for BSE and the culling of feed and age cohorts of BSE cases. The food risk in the baseline scenario ranged from 16.98 lost life years in 2002 to 2.69 lost life years in 2005. Removing specified risk materials removal practices, post-mortem testing and post-mortem testing plus cohort culling reduced this risk with 93%, 82.7% and 83.1%. The estimated cost-effectiveness of all BSE measures in The Netherlands ranged from 4.3 million euros per life year saved in 2002 to 17.7 million euros in 2005. It was discussed that the cost-effectiveness of BSE control strategies will further deviate from regular health economics thresholds as BSE prevalence and incidence declines.

Epidemiological investigations into the sources of Salmonella contamination of pork.

This study was conducted to elucidate which phases of the pork production chain contribute to the Salmonella contamination on pork after slaughter. During 7 sampling days, samples were collected of randomly selected slaughter pigs and of pigs from selected Salmonella-infected and Salmonella-free herds, trucks, lairages, and slaughterlines, in two slaughterhouses. Salmonella genotypes, present on pork after slaughter, were compared with Salmonella types, present on the farm, in the truck, in the lairage, on slaughter equipment, and in pigs from other herds. Results showed that the slaughterline was the most important source of Salmonella contamination of carcasses. The farm was the most important source of contamination of livers, tongues, rectal samples and mesenterial lymphnodes, for pigs originating from sero-positive herds. The lairage was the most important contamination source for pigs originating from sero-negative herds, for all samples, except carcasses. It is recommended to avoid each direct or indirect contact between different herds along the whole pork production chain, especially between Salmonella-infected and Salmonella-free herds.