Ann Lindberg

Principles for eradication of bovine viral diarrhoea virus (BVDV) infections in cattle populations.

Systematic eradication of BVDV without vaccination started in Scandinavia in 1993. In principle, the schemes include; (1) identification of non-infected and infected herds using different combinations of serological herd tests such as bulk milk tests and spot tests (sample of animals in a certain age), (2) monitoring/certification of non-infected herds by repeated sampling, applying one of the above-mentioned methods and (3) virus clearance in infected herds aimed at removing persistently infected (PI) animals in a cost- and time-efficient manner. In the virus clearance protocol described, an initial test is performed on all animals with subsequent follow-up of calves born as well as of dams seronegative in the initial test. It is generally recommended to perform an initial antibody test on all samples. This should be done not only to screen for seronegative animals on which virus isolation should be attempted (i.e. possible PI animals), but more in order to identify non-immune animals in reproductive age, that is, the key animals in herd-level persistence of infection. In Sweden, a common finding has been self-clearance, where the infection ceases without any other intervention than controlled introduction of new animals. Other epidemiological observations concern the course of events following virus introduction. Important risk factors for spreading BVDV are discussed, where livestock trade is perceived as the most central to control. Live vaccines, imported semen and embryos constitute special hazards, since they may act as vehicles for the introduction of new BVDV strains. The importance of making farmers aware of herd biosecurity and their own responsibility for it is stressed, and in order to maintain a favourable situation after a scheme has been concluded, effort must be put into establishing such a persisting attitude in the farming community.

Molecular typing of isolates of Clostridium perfringens from healthy and diseased poultry

The bacterium Clostridium perfringens can cause both clinical and subclinical disease in poultry. To study the pathogenesis and epidemiology of disease caused by C. perfringens, methods for typing its various strains need to be evaluated. C. perfringens isolates from healthy and diseased poultry from different parts of Sweden were analysed by polymerase chain reaction (PCR) in order to establish the presence of alpha-, beta-, beta2-, epsilon -, iota- and enterotoxin genes. In order to subtype C. perfringens isolates, the two methods amplified fragment length polymorphism (AFLP) and pulsed field gel electrophoresis (PFGE) were compared on 21 C. perfringens isolates from 10 different farms. In a second study, 32 isolates of C. perfringens type A from three broilers from a healthy flock reared without ionophorous anticoccidials were subtyped by PFGE. All 53 isolates analysed with PCR belonged to the toxin type A of C. perfringens, with the gene coding for alpha-toxin production. Two isolates possessed the beta2-gene as well, but none had the other toxin genes. Both AFLP and PFGE differentiated 21 strains into 10 different subtypes. This differentiation correlated closely with the origins of the isolates. Unique subtypes were isolated from seven farms. Only isolates from birds of one farm demonstrated more than one subtype of C. perfringens. The subtyping of the isolates from a healthy flock showed that each bird carried two to three different subtypes and two different subtypes were found in the same kind of tissue sample in four cases. Three of the four different subtypes found in this study were new, compared with the first study. AFLP and PFGE were found to be equally suitable for subtyping of C. perfringens isolates. The wide variation in subtypes in the healthy broilers could be the result of the antibiotic-free rearing of these birds.

Microbial Aetiology of Acute Clinical Mastitis and Agent-Specific Risk Factors

A nation wide study on the microbial aetiology of cases of acute clinical mastitis in Swedish dairy cows was conducted with the aim to investigate changes in the microbial panorama compared to a previous study performed 1994-1995. Another aim was to investigate some agent-specific environmental and individual risk factors. Milk samples were collected from 987 udder quarter cases from 829 cows during six 2-month periods from May 2002 to April 2003, and data on risk factors and demography were collected at sampling by means of a questionnaire. In total, 1056 bacteriological diagnoses were made. The most frequently isolated bacterial species was Staphylococcus aureus constituting 21.3% of the diagnoses, followed by Escherichia coli (15.9%), Streptococcus dysgalactiae (15.6%), Streptococcus uberis (11.1%), coagulase-negative staphylococci (6.2%), Arcanobacterium pyogenes (6.1%) and Klebsiella spp. (4.2%). Samples with no growth or contamination constituted 10.6% and 4.5% of the diagnoses, respectively. A major shift in the panorama of udder pathogens was not observed compared to the survey in 1994-1995. Isolation of Klebsiella spp. was strongly associated with the use of sawdust as bedding material. On the other hand, using sawdust as bedding reduced the risk of isolating S. uberis relative to using straw or peat. The risk of isolating E. coli increased with increasing milk yield and was higher in loose housing systems than in tie stalls. Isolation of S. aureus was associated with tie stalls, and A. pyogenes with low yielding cows and teat lesions. S. dysgalactiae infections were also associated with teat lesions.

Network analysis of cattle and pig movements in Sweden : Measures relevant for disease control and risk based surveillance

Registration of cattle and pig movements is mandatory in Sweden and all registered movements between farms in the years 2006-2008 were investigated using network analysis. The networks were analysed as monthly and yearly networks, separately per species and with the two species together. Measures that have been previously discussed in relation to outbreaks and disease control were calculated; moreover a measure of the ingoing infection chain was constructed. The ingoing infection chain captures ingoing contacts through other holdings, taking the temporal aspect and sequence of the movements into account. The distribution of the contacts among the holdings was skewed. Many farms had few or no contacts, while others had many, a pattern which has also been described from other countries. The cattle network and the combined network showed a recurring seasonal pattern, while this was not seen in the pig network. The in-degree was not equivalent to the ingoing infection chain; there were holdings with limited direct contacts, but a large number of indirect contacts. The ingoing infection chain could be a useful measure when setting up strategies for disease control and for risk based surveillance as it identifies holdings with many contacts through live animal movements and thus at potentially higher risk for introduction of contagious diseases.

Validation of a Neospora caninum iscom ELISA without a gold standard.

Neospora caninum is an intracellular parasite which causes abortion in cattle worldwide. One problem in the validation of the different methods for demonstration of this parasite is the lack of an appropriate gold standard. To validate an immunostimulating complex (iscom) enzyme-linked immunoassay (ELISA) used to detect antibodies to N. caninum, sera from 244 cattle in five Swedish dairy herds infected with N. caninum were analysed. The sera also were analysed by a standard indirect-fluorescent antibody test (IFAT). The results obtained by the two tests were compared using the Gibbs sampler. Gibbs sampling is a latent-class approach based on Bayesian statistics; neither test is assumed to be more correct in stating the true status of infection. The Gibbs sampler was run using both informative and non-informative prior probabilities. We also simulated different cut-offs in the iscom ELISA (providing data to inform selection of optimal cut-off values for different applications). The ELISA produced fewest incorrect test results over all at a cut-off value of 0.200. The sensitivity and specificity at this cut-off were 99 and 96%, respectively. The IFAT had a high specificity (99%) but a lower sensitivity (78%) than expected-confirming that the IFAT cannot be treated as a true gold standard. Sensitivity and specificity of the ELISA were presented in a two-graph receiver operating characteristic (TG-ROC) plot. Any cut-off between 0.150 and 0.300 will have both sensitivity and specificity > or =95%. Optical densities of < or =0.150 and > or =0.550 (or > or =0.350) were suggested as limits to rule out and rule in infection, respectively.

Failure to spread bovine virus diarrhoea virus infection from primarily infected calves despite concurrent infection with bovine coronavirus.

Abstract Previous reports on the spread of bovine virus diarrhoea virus (BVDV) from animals primarily infected with the agent are contradictory. In this study, the possibility of transmission of BVDV from calves simultaneously subjected to acute BVDV and bovine coronavirus (BCV) infection was investigated. Ten calves were inoculated intranasally with BVDV Type 1. Each of the 10 calves was then randomly allocated to one of two groups. In each group there were four additional calves, resulting in five infected and four susceptible calves per group. Virulent BCV was actively introduced in one of the groups by means of a transmitter calf. Two calves, susceptible to both BVDV and BCV, were kept in a separate group, as controls. All ten calves actively inoculated with BVDV became infected as shown by seroconversions, and six of them also shed the virus in nasal secretions. However, none of the other eight calves in the two groups (four in each) seroconverted to this agent. In contrast, it proved impossible to prevent the spread of BCV infection between the experimental groups and consequently all 20 study calves became infected with the virus. Following infection, BCV was detected in nasal secretions and in faeces of the calves and, after three weeks in the study, all had seroconverted to this virus. All calves, including the controls, showed at least one of the following clinical signs during days 3–15 after the trial started: fever (≥40°C), depressed general condition, diarrhoea, and cough. The study showed that BVDV primarily infected cattle, even when co-infected with an enteric and respiratory pathogen, are inefficient transmitters of BVDV. This finding supports the principle of the Scandinavian BVDV control programmes that elimination of BVDV infection from cattle populations can be achieved by identifying and removing persistently infected (PI) animals, i.e. that long-term circulation of the virus without the presence of PI animals is highly unlikely.

Comparison between dairy cow disease incidence in data registered by farmers and in data from a disease-recording system based on veterinary reporting

Abstract Sweden has a national disease-recording system based on veterinary reporting. From this system, all cattle-disease records are transferred to the dairy industry cattle database (DDD) where they are used for several purposes including research and dairy-health statistics. Our objective was to evaluate the completeness of this data source by comparing it with disease data registered by dairy farmers. The proportion of veterinary-treated disease events was estimated, by diagnosis. Disease incidence in the DDD was compared, by diagnosis and age, with disease data registered by the farmers. Comparison was made, by diagnosis, for (i) all disease events and (ii) those reported as veterinary-treated. Disease events, defined as “observed deviations in health, from the normal” were recorded by the farmers during January, April, July and October 2004. For the diagnoses calving problems, peripartum disorders, puerperal paresis and retained placenta, incidence proportions (IP) with 95% confidence intervals (CIs) were estimated. For all other disease problems, incidence rates (IR) were used. In total, 177 farmers reported at least 1 month and 148 reported all 4 months. Fifty-four percent of all disease events in the farmers’ data were reported as veterinary-treated. For several of the most common diagnoses, the IRs and IPs for all events were significantly higher in farmers’ data than in the DDD. Examples are, in cows: clinical mastitis, cough, gastro-intestinal disorders and lameness in hoof and limb; and in young stock: cough and gastro-intestinal disorders. For veterinary-treated events only, significant differences with higher IR in the farmers’ data were found in young stock for sporadic cough and sporadic gastro-intestinal disorders. The diagnosis “other disorders” had significantly more events in the DDD than in farmers’ data, i.e. veterinarians tended to choose more unspecific diagnoses than the farmers. This result indicates that the true completeness is likely to be higher than our estimate. We conclude that for the time period studied there was differential under-reporting associated with the diagnosis, the age of the animal and whether the herd was served by a state-employed or private veterinarian.

Validation of a test for dams carrying foetuses persistently infected with bovine viral-diarrhoea virus based on determination of antibody levels in late pregnancy.

Our objective was to estimate, using a generalised linear mixed-model approach, the sensitivity and specificity of an indirect ELISA when used to identify dams pregnant with persistently bovine viral-diarrhoea virus (BVDV)-infected foetuses. Cows that had been tested for antibodies to BVDV with a positive result during their pregnancy and where the offspring had been tested for both antibody and virus were identified by accessing the Swedish BVD database and the official pedigree records. The resulting data set consisted of 2162 cow-calf pairs in 126 herds, of which 281 included virus-positive calves. The sensitivities and specificities at 12 different decision thresholds (corresponding to optical densities (ODs) between 0.5 and 1.6) were estimated using generalised linear mixed models (binomial error, logit link), in which the gold standard (the BVDV status of the calf) was included as a covariate. In each model, the dependent variable was the dichotomous test result at the decision threshold in question. There was a significant positive interaction between the calfs status and gestational stage in all 12 models--indicating that the sensitivity and specificity at any given decision threshold was improved when the the test was performed later in pregnancy. The test should be applied only when samples have been taken in late gestation--not before the seventh month in pregnancy. If applied during the last months of pregnancy, the point estimate of the sensitivity ranges between 0.94 and 1.0 as the decision threshold is moved from 1.0 and downwards to 0.7. Similarly, the specificity ranges between 0.39 and 0.67 as the decision threshold is moved from 0.8 and upwards to 1.1.

Risk factors for calf mortality in large Swedish dairy herds.

Abstract The aim of this study was to identify possible risk factors for 1–90 day calf mortality in large Swedish dairy herds. Sixty herds with a herd size of ≥160 cows were visited once between December 2005 and March 2006. Thirty herds were known to have low mortality (LM) and 30 were known high mortality herds (HM). Upon the visit, data about housing and management was collected from interviews with personnel responsible for the calves. The herd status regarding the calves’ passive transfer (total protein), levels of α-tocopherol, β-carotene and retinol, and excretion of faecal pathogens (Cryptosporidium spp., Escherichia coli F5, rota and corona virus) was evaluated based on targeted sampling of high risk calf groups; in each herd, blood and faecal samples were collected from calves 1–7 and 1–14 days old, respectively. Similarly, the herd status regarding clinical respiratory disease in calves and history of respiratory virus exposure was evaluated based on lung auscultations and blood samplings of calves 60–90 days old. The median calf mortality risk (in calves 1–90 days of age) among HM herds was 9% (Range: 6–24%) and among LM herds 1% (Range: 0–2%). LM and HM herds were compared using five logistic regression models, covering potential risk factors within different areas: “Disease susceptibility”, “Factors affecting the gastrointestinal tract”, “Factors related to transmission of infectious disease”, “Hygiene” and “Labour management”. The percentage of calves, 1–7 days old, with inadequate serum concentrations of α-tocopherol and β-carotene were significantly higher in HM herds compared to LM herds and also associated with higher odds of being a HM herd (OR=1.02; p =0.023 and OR=1.05; p =0.0028, respectively). The variable “Average number of faecal pathogens in the sampled target group” was significantly associated with higher odds of being a HM herd (OR=4.65; p =0.015), with a higher average in HM herds. The percentage of calves with diarrhoea treated with antibiotics was significantly higher in HM herds and was associated with higher odds of being a HM herd (OR=1.08; p =0.021). The median age at death of calves in the age interval 1–90 days that died during a one-year period was significantly lower among HM herds (13 days) than in LM herds (24 days) (p =0.0013) The results indicate that gastrointestinal disorders may be an important cause of calf mortality in large Swedish dairy herds. Furthermore, our study provides additional indications that fat soluble vitamins might play an important role for calf health.

Incidence of mastitis and bacterial findings at clinical mastitis in Swedish primiparous cows - influence of breed and stage of lactation.

Mastitis is a common disease also among primiparous dairy cows. Identification of the extent and type of problem is important to initiate correct control measures. In Sweden, unique national production and disease databases are available. The main aim of the study was to investigate the occurrence of mastitis, measured by the annual incidence of veterinary-treated clinical mastitis (VTCM) and geometric mean of monthly milk somatic cell count (SCC) recordings in Swedish primiparous cows in relation to older cows during 2002-2006 with emphasis on breed differences. Other aims were to study differences between primiparous and older cows in the distribution of bacterial findings at clinical mastitis, and the occurrence of VTCM and bacterial findings in relation to stage of lactation using data from a Swedish field study performed 2002-2003. Descriptive statistics and univariable analyses were used in the investigations. During 2002-2006 approximately 10% of Swedish primiparous cows experienced VTCM each year, while the geometric mean SCC of Swedish primiparous cows was approximately 65,000 ml(-1). Both parameters were lower than in older cows. Primiparous cows of the Swedish Red (SR) breed had better udder health than cows of the Swedish Holstein (SH) breed. The overall distribution of udder pathogens was similar in primiparous and older cows. In primiparous cows, most VTCM occurred during the first week after calving, and Staphylococcus aureus and Streptococcus dysgalactiae were the most common udder pathogens during this period. Better control measures directed at these infections are warranted around calving to reduce the risk of mastitis in primiparous cows.