Lis Alban

Longitudinal Study of Salmonella enterica Serotype Typhimurium Infection in Three Danish Farrow-to-Finish Swine Herds

ABSTRACT A longitudinal study of the infection dynamics of Salmonella enterica was carried out with three Danish farrow-to-finish swine herds. To account for variations in Salmonella shedding over time, litters from each herd were divided into two cohorts. Each cohort consisted of 30 pigs, for a total of 180 pigs. Pigs were individually monitored by monthly bacteriologic and serologic examinations from weaning to slaughter. At weaning, individual sows were examined bacteriologically and serologically. At slaughter, cecal contents, ileocecal lymph nodes, and carcass swab samples were obtained from 131 pigs. A total of 88 pigs were found to be shedding Salmonella on one or more occasions. Only the Salmonella serotype Typhimurium was detected during the study period. At weaning, no sows or piglets were found to be shedding, but a serological reaction was detected in 11 sows. The prevalence in culture peaked in the nursery and subsequently declined to undetectable levels before slaughter. The seroprevalence peaked approximately 60 days after the peak prevalence in culture. Salmonella was detected in individual fecal samples at least once in 53% of the pigs, and 62% of the pigs were seropositive more than once. Only 3.7% of all pigs were found to be culture positive on more than one occasion. Piglets from seroreacting sows had a significantly (P = 0.0339) lower probability of shedding in the nursery. Under the assumption that shedding lasted at least 1 or 2 weeks, the average shedding time was estimated to have been 18 or 26 days. An association between serology, on-farm bacteriology, and Salmonella prevalence in culture at slaughter was shown. Marked differences in prevalence in sera and prevalence in culture between cohorts and within herds were observed. These differences emphasize the need for caution when using point estimates in on-farm interventions and surveillance in subclinically infected swine herds.

Lameness in Danish dairy cows: frequency and possible risk factors

A retrospective longitudinal study was conducted aiming at determining specific risk factors associated with lameness in lactating dairy cattle. A total of 9762 cows from 165 Danish dairy herds was included in the analysis. The evaluated factors were: breed and parity of the cow, season in which calving took place, whether the cow had reproductive diseases, udder-related diseases, or metabolic/digestive diseases during the lactation, herd size, the farmers expectation of whether he would still be a milk producer 5 years ahead, whether the cows were on pasture in the summer, housing system, and way of using veterinarian. The association between the possible risk factors and lameness was assessed using logistic-binomial regression with herd as a random effect. The results suggest that a cow has an increased risk of developing lameness during a lactation if she is either a Danish Black and White, a Red Danish or a Danish Red and White (compared with Danish Jersey), if she is in her first or fourth lactation, if she has calved during the housing period (October–April), if the farmer does not expect to be a dairy producer 5 years ahead, and if she is part of a large herd.

Comparing the epidemiological and economic effects of control strategies against classical swine fever in Denmark

In 2006, total Danish pork exports were valued at 3.8 billion euros, corresponding to approximately 5% of the total Danish exports, and an outbreak of a notifiable disease would have dramatic consequences for the agricultural sector in Denmark. Several outbreaks of classical swine fever (CSF) have occurred in Europe within the last decade, and different control strategies have been suggested. The objective of this study was to simulate the epidemiological and economic consequences of such control strategies in a CSF epidemic under Danish conditions with respect to herd demographics and geography and to investigate the effect of extra biosecurity measures on farms. We used InterSpread Plus to model the effect of nine different control strategies: the minimum measures required by the EU plus depopulation of contact herds (EUplus), extra depopulation of neighbouring herds, extra surveillance within the protection and surveillance zones, extra biosecurity in SPF herds-or in all herds, vaccination of all pigs in the 1 or 2 km zones using live vaccine as a protective measure (vaccination-to-kill), vaccination of all weaners and finishers in the 1 or 2 km zones using an E2 marker vaccine as a suppressive measure (vaccination-to-live). Each epidemic was simulated to start in four different index herds: production herds located in low, medium and high pig density areas, respectively; and a nucleus herd in an area of high pig density. For each control strategy and index case, we calculated the size and duration of the epidemic, the number of depopulated and/or vaccinated herds and animals, the control costs borne by the public and the pig industry, respectively, as well as the loss of exports associated with the epidemic. The simulations showed that the EUplus strategy is the most effective of the evaluated strategies with respect to limiting the size, duration and cost of the epidemic, regardless of the index case. However, regarding the number of slaughtered animals, the vaccination-to-live strategies appeared to be more effective. Epidemics become larger and last longer if the index case is a nucleus herd. This implies that biosecurity in nucleus herds is extremely important to avoid transmission of CSF to these herds. Simulations showed that a Danish CSF epidemic will be moderate in most cases and will include fewer than 10 cases and last less than 2 weeks on average. However, for some iterations, long-lasting and large epidemics were observed. Irrespective of the size and duration, an epidemic is expected to be very costly due to the export losses.

Prevalence and characteristics of epilepsy in the Belgian shepherd variants Groenendael and Tervueren born in Denmark 1995-2004.

BackgroundThe Belgian shepherd Groenendael and Tervueren is believed to be at higher risk of developing epilepsy than dogs of the common population. This epidemiological study was designed to estimate the prevalence of epilepsy in the Danish population of Groenendael and Tervueren born between 1995 and 2004. Furthermore, it was the intention to describe the clinical manifestation (seizure types and phenomenology) of epilepsy and to identify risk factors for euthanasia once the dog was diagnosed as having epilepsy.MethodsAll owners of Groenendael and Tervueren dogs born between January 1995 and December 2004 and registered in the Danish Kennel Club (1,248 dogs) were contacted and asked to answer a mailed questionnaire concerning epilepsy. Positive responders were subsequently validated in a follow-up interview conducted by telephone using a standardized questionnaire. Owners were questioned about age at first seizure, seizure frequency, seizure duration, a detailed description of seizure phenomenology, post-ictal signs and if a veterinarian had diagnosed the dog with epilepsy.ResultsPrevalence of epilepsy was estimated at 9.5%. Mean age of epilepsy debut was 3.3 years (range 0.5–8.0 years). There was an almost equal number of Groenendael (25) and Tervueren (24). The distribution of females and males was 31 and 18 respectively. Twenty-five per cent experienced focal seizures, 53% experienced focal seizures with secondary generalization and 18% experienced primary generalized seizures. In four percent seizures were unclassifiable. The most commonly reported focal seizure phenomenology included ataxia, crawling, swaying, fearful behavior, salivation, excessive attention seeking and disorientation. In 16% of the cases, epilepsy led to euthanasia. Intact dogs with epilepsy had a significantly increased risk of being euthanized because of epilepsy compared to neutered dogs with epilepsy. In 22% of the cases the owners reported that anxiety/hyperactivity/stress could act as a seizure provoking factor.ConclusionA high prevalence of epilepsy appears to be present in the Danish Groenendael and Tervueren population. The relatively late debut age of epilepsy in this breed contributes greatly to the increased prevalence of epileptic individuals, because dogs developing epilepsy late in life are used for breeding unintended.

Qualitative and quantitative risk assessment for human salmonellosis due to multi-resistant Salmonella Typhimurium DT104 from consumption of Danish dry-cured pork sausages

Salmonella Typhimurium DT104 (DT104) is unwanted in products for human consumption due to its antibiotic resistance and ability to cause disease. We intended to set up an improved monitoring and management program to aid in deciding when to use pork contaminated with DT104 for production of sausages without jeopardizing consumer safety. We started by carrying out two assessments of the risk for human health associated with consumption of sausages produced by: (1) Danish pork from average slaughter days; (2) imported pork (IMP) with average prevalence of DT104. The assessments showed that, if Salmonella is present, it is usually in lower numbers (< or =50 per 400 cm(2) surface). Additionally, during processing, the numbers will be reduced by at least 2 log-units. In Danish (DK) pork, DT104 constitutes 0.2-1.0% of the Salmonella isolates reported, while in imported pork (IMP), 18%. We estimated that out of one million, 25 g servings of DK dry-cured sausages, up to two DT104 bacteria could be found in each of 245 servings. Out of one million servings of 25 g IMP dry-cured sausages, up to two DT104 bacteria would occur in each of 19,260 servings.

Simulating the spread of classical swine fever virus between a hypothetical wild-boar population and domestic pig herds in Denmark.

Denmark has no free-range wild-boar population. However, Danish wildlife organizations have suggested that wild boar should be reintroduced into the wild to broaden national biodiversity. Danish pig farmers fear that this would lead to a higher risk of introduction of classical swine fever virus (CSFV), which could have enormous consequences in terms of loss of pork exports. We conducted a risk assessment to address the additional risk of introducing and spreading CSFV due to the reintroduction of wild boar. In this paper, we present the part of the risk assessment that deals with the spread of CSFV between the hypothetical wild-boar population and the domestic population. Furthermore, the economic impact is assessed taking the perspective of the Danish national budget and the Danish pig industry. We used InterSpreadPlus to model the differential classical swine fever (CSF) risk due to wild boar. Nine scenarios were run to elucidate the effect of: (a) presence of wild boar (yes/no), (b) locations for the index case (domestic pig herd/wild-boar group), (c) type of control strategy for wild boar (hunting/vaccination) and (d) presence of free-range domestic pigs. The presence of free-range wild boar was simulated in two large forests using data from wildlife studies and Danish habitat data. For each scenario, we estimated (1) the control costs borne by the veterinary authorities, (2) the control-related costs to farmers and (3) the loss of exports associated with an epidemic. Our simulations predict that CSFV will be transmitted from the domestic pig population to wild boar if the infected domestic pig herd is located close to an area with wild boar (<5 km). If an outbreak begins in the wild-boar population, the epidemic will last longer and will occasionally lead to several epidemics because of periodic transfer of virus from groups of infected wild boar to domestic pig herds. The size and duration of the epidemic will be reduced if there are no free-range domestic pig herds in the area with CSF-infected wild boar. The economic calculations showed that the total national costs for Denmark (i.e. the direct costs to the national budget and the costs to the pig industry) related to an outbreak of CSF in Denmark will be highly driven by the reactions of the export markets and in particular of the non-EU markets. Unfortunately, there is a substantial amount of uncertainty surrounding this issue. If hunting is used as a control measure, the average expenses related to a CSF outbreak will be 40% higher if wild boar are present compared with not present. However, a vaccination strategy for wild boar will double the total costs compared with a hunting strategy.

Epidemiologic and economic evaluation of risk-based meat inspection for bovine cysticercosis in Danish cattle.

Under the current EU meat inspection regulation, every single carcase from all bovines above 6 weeks of age has to be examined for bovine cysticercosis (BC). This is time-consuming, costly, and is of limited value in countries with low prevalence. The aim of this study was to develop a stochastic simulation model for analysis of tentative risk-based meat inspection systems for BC in Danish cattle with regard to system sensitivity (SSSe), specificity and potential monetary benefits compared to the current system, which has an estimated SSSe of 15%. The relevant risk factors used to construct three alternative scenario trees were identified from previous Danish risk factor studies (1) gender, (2) grazing and (3) access to risky water sources. Thus, females, animals that had been grazing or animals with access to risky water sources were considered high-risk and would be subjected to invasive inspection at meat inspection. All animals in the low-risk groups (i.e. males, non-grazing or no access to risky water sources, respectively) would be subjected to visual inspection only. It was assumed that half of the cattle were slaughtered in abattoirs that would be able to reorganise the work at the slaughterline, allowing them to do with one meat inspector less. All abattoirs would gain on the price of sold uncut beef from the masseter muscles from visually inspected cattle. Under these assumptions, using gender and grazing were preferable due to them having SSSe only slightly lower than the current system, and highest effectiveness ratios, but they had a lower net economic effect (NEE) than the scenario using risky water sources. Using gender to differentiate high and low-risk groups was judged preferable over grazing due to feasibility, because the information is readily available at the slaughter line. The exact total NEE for the cattle sector depends on how many and which of the abattoirs that would be able to reorganise the work at the slaughter line to save money on inspection of the head of carcases. Overall, the SSSe was low in all scenarios leading to undetected BC-positive cattle both in the current meat inspection and under the investigated risk-based meat inspection systems. Therefore, improving the sensitivity of the methods used for inspection of high-risk cattle would be beneficial.

Occurrence and factors associated with bovine cysticercosis recorded in cattle at meat inspection in Denmark in 2004-2011.

Current EU regulation requires that every bovine carcass is examined for bovine cysticercosis (BC) at meat inspection. This is costly and might be superfluous at low BC prevalence. However, from a consumer view-point it may be important to identify and manage infected carcasses to avoid human infection. If relevant data could be effectively used to classify animals with respect to their risk of being infected, then the current meat inspection could be replaced by a more cost-effective system targeting high-risk animals. This study aimed to (1) describe the distribution of BC cases in the Danish cattle population, (2) estimate the animal level prevalence (3) provide descriptive statistics of potential risk factors for BC, and (4) determine attributable risks and fractions of selected risk factors potentially useful for a future risk-based meat inspection system. In total, 348 cases of BC were recorded among all cattle slaughtered (n=4,090,661) in Denmark between 2004 and 2011. The true animal level prevalence of BC was estimated to be 0.06%. The herd of origin of the cases were defined as the herd in which the animals spent most of their lifetimes. The detected cases were found to originate from 328 herds, with a maximum of two cases per herd indicating sporadic occurrence. Even though organic farming was associated with a higher risk (RR=1.9 in univariable analysis) of BC-positive animals being detected at slaughter, the population attributable fraction showed that only 5% of the animals with BC could be attributed to organic farming practices at the level of organic farming practiced in Denmark in the study period. Thus, organic farming status was not a suitable risk factor to use to target future risk-based meat inspection. However, 54% of the animals with BC in the cattle population were attributed to female gender. Increasing age at slaughter was also associated with high risk of BC. There may be overlaps between these effects in animals with multiple risk factors. Other underlying factors such as grazing patterns might explain the risk factors and attribution results found in this study. However, grazing practices are currently not recorded in the Danish cattle database. Therefore, animal level risk factors such as age and gender together with other risk factors such as grazing practices might be included as food chain information, required to be provided by the farmer prior to slaughter. The challenges and opportunities of this approach should be investigated further.

Factors affecting persistence of high Salmonella serology in Danish pig herds

In 1995, Denmark implemented a National Surveillance-and-Control Programme for Salmonella in finisher pigs. It was based on serological surveillance that enabled assignment of pig herds into three prevalence levels. Herds in levels 2 and 3 (corresponding to moderate and high Salmonella seroprevalence) were subjected to control measures, compulsory pen faecal sampling for bacteriological examination and serotyping. The initial dataset contained all herds that had mandatory bacteriological samples taken (N=3691 samples), out of which 44.4% were negative. Data from January 2005 to November 2007 obtained from the Danish programme were used to investigate the persistence of high serology (Salmonella index > or =40, corresponding to levels 2 or 3) for herds infected with one of the four most common Salmonella serotypes found in pig herds, S. Typhimurium, S. Derby, S. Infantis or S. Livingstone, using survival analysis. The effects of other potential factors, such as season, year, herd size and herd health status were also evaluated. An event was defined as return to low serology measured by a Salmonella index <40, for at least 6 consecutive months. A total of 1242 pig herds positive for one of the four serotypes of interest and 1433 events were included in the preliminary analysis using the Kaplan-Meyer method. The median high serology time (survival time) was 8 months and all explanatory variables were found to be significant (p<0.05), except the herd health status (p=0.49). Cox proportional hazards regression was used for herds infected with S. Typhimurium (N=1066) or S. Derby (N=314). Herds infected with S. Typhimurium presented a lower probability for returning to low serology (HR=0.69, p<0.001), compared to S. Derby, indicating longer high serology periods. Herds that started having high seroprevalence in Summer and Fall had a higher probability of returning to low serology compared to those starting in Winter and Spring (p<0.001). Compared to 2005, periods with high Salmonella serology were fewer and shorter in 2006 and 2007 (p<0.001). A positive impact of herd size was found, with larger herds being associated with shorter high serology periods, compared to smaller herds (p=0.012). This might indicate effective implementation of biosecurity measures in large herds. The health status of the herds was not found to be a significant factor (p=0.87), which might reflect that biosecurity measures in place in SPF herds are not effective for clearing Salmonella infections.

Exposure assessment of extended-spectrum beta-lactamases/AmpC beta-lactamases-producing Escherichia coli in meat in Denmark

Introduction Extended-spectrum beta-lactamases (ESBL) and AmpC beta-lactamases (AmpC) are of concern for veterinary and public health because of their ability to cause treatment failure due to antimicrobial resistance in Enterobacteriaceae. The main objective was to assess the relative contribution (RC) of different types of meat to the exposure of consumers to ESBL/AmpC and their potential importance for human infections in Denmark. Material and methods The prevalence of each genotype of ESBL/AmpC-producing E. coli in imported and nationally produced broiler meat, pork and beef was weighted by the meat consumption patterns. Data originated from the Danish surveillance program for antibiotic use and antibiotic resistance (DANMAP) from 2009 to 2011. DANMAP also provided data about human ESBL/AmpC cases in 2011, which were used to assess a possible genotype overlap. Uncertainty about the occurrence of ESBL/AmpC-producing E. coli in meat was assessed by inspecting beta distributions given the available data of the genotypes in each type of meat. Results and discussion Broiler meat represented the largest part (83.8%) of the estimated ESBL/AmpC-contaminated pool of meat compared to pork (12.5%) and beef (3.7%). CMY-2 was the genotype with the highest RC to human exposure (58.3%). However, this genotype is rarely found in human infections in Denmark. Conclusion The overlap between ESBL/AmpC genotypes in meat and human E. coli infections was limited. This suggests that meat might constitute a less important source of ESBL/AmpC exposure to humans in Denmark than previously thought – maybe because the use of cephalosporins is restricted in cattle and banned in poultry and pigs. Nonetheless, more detailed surveillance data are required to determine the contribution of meat compared to other sources, such as travelling, pets, water resources, community and hospitals in the pursuit of a full source attribution model.