Bovine natural killer cells

Natural killer (NK) cells have received much attention due to their cytotoxic abilities, often with a focus on their implications for cancer and transplantation. But despite their name, NK cells are also potent producers of cytokines like interferon-gamma. Recent discoveries of their interplay with dendritic cells and T-cells have shown that NK cells participate significantly in the onset and shaping of adaptive cellular immune responses, and increasingly these cells have become associated with protection from viral, bacterial and parasitic infections. Furthermore, they are substantially present in the placenta, apparently participating in the establishment of normal pregnancy. Consequently, NK cells have entered arenas of particular relevance in veterinary immunology. Limited data still exist on these cells in domestic animal species, much due to the lack of specific markers. However, bovine NK cells can be identified as NKp46 (CD335) expressing, CD3(−) lymphocytes. Recent studies have indicated a role for NK cells in important infectious diseases of cattle, and identified important bovine NK receptor families, including multiple KIRs and a single Ly49. In this review we will briefly summarize the current understanding of general NK cell biology, and then present the knowledge obtained thus far in the bovine species.     

Determination of the sequence of the complete open reading frame and the 5'NTR of the Paderborn isolate of classical swine fever virus

The classical swine fever (CSF) epidemic in the Netherlands in 1997-1998 lasted 14 months, during which 429 infected and 1300 at risk herds were culled, at an estimated economical cost of 2 billion US dollars. Despite the overwhelming scale of the epizootic, the CSF virus (CSFV) strain causing the outbreak has remained largely uncharacterized. The Dutch epizootic is epidemiologically linked to a small CSF outbreak in 1997, in Paderborn in Germany. E2 and partial 5' NTR sequencing has shown that the index Paderborn isolate, and several Dutch isolates taken during the 1997-1998 epizootic, are virtually identical, confirming that the Paderborn isolate triggered the Dutch outbreak, and furthermore showing that this single isolate was stable throughout the whole Dutch outbreak (the above reviewed in [C. Terpstra, A. J. de Smit, Veterinary Microbiol. 77 (2000) 3-15]). We determined the nucleotide sequence of the 5' NTR (by 5' RACE) and the complete open reading frame of the Paderborn isolate (GenBank AY072924). Our sequence was identical to previously published partial 5'NTR and E2 sequences for the index Paderborn 1997 and Dutch 1997 (Venhorst) isolates, confirming the identity of the virus we sequenced. Phylogenetic analysis based on the complete open reading frame showed that Paderborn is genetically very different from common European laboratory reference strains. Neutralization studies showed that Paderborn is also antigenically very different from common laboratory strains such as Alfort 187. Paderborn is the only recent European CSFV field isolate for which a complete sequence is available, and given Paderborns genetic and antigenic uniqueness, the Paderborn sequence may have practical use for diagnostic and vaccine antigen development.     

Data-quality issues and alternative variable-screening methods in a questionnaire-based study on subclinical Salmonella enterica infection in Danish pig herds

Our aim was to determine risk factors for subclinical Salmonella enterica infection in Danish finishing-pig herds. In this paper, the evaluation, combining and initial reduction of variables is presented, along with assessment of the hypotheses in the preliminary statistical testing.The first group of herds was selected at random with no former knowledge of S. enterica infection. Both the herd prevalence and the within-herd prevalence among these herds turned out to be low; hence, some additional herds were selected from The Danish Salmonella Control program, based on their high seroprevalence. This resulted in a hybrid case-"control" design of the study and therefore, five different methods of categorising the data were used to ensure that variables were not wrongfully excluded as a result of using an improper design.Our questionnaire focused on management, infection-limiting precautions and feed and feeding procedures. To establish the prevalence of S. enterica infection within herds at the time of the visit, 50 blood samples from each herd were collected and serologically examined. The reliability of each variable from the questionnaire was assessed and it was decided which variables should be selected, disregarded, combined with other variables and/or recoded. In the simple statistical testing (2x2 tables, cut-off: P=0.25) herds were defined as subclinically S. enterica infected if the within-herd proportion of individual pigs with OD%>10 was more than 20%.The results included questionnaires from 96 randomly selected and 39 high-seroprevalence herds and 6814 blood samples. The initial 95 variables originally included in the questionnaires were reduced to 21 by critical check, combination, recoding and preliminary screening. We failed to demonstrate "herd size" as a risk factor for subclinical S. enterica infection in pig herds.     

Monitoring porcine reproductive and respiratory syndrome virus infection status in swine herds based on analysis of antibodies in meat juice samples

An indirect ELISA test was developed as a novel tool aimed at monitoring the herd infection status of swine herds. Meat juice samples from pig carcasses were analysed for the presence of antibodies against porcine reproductive and respiratory syndrome virus (PRRSV). A study of samples from herds with known PRRS status was undertaken. The PRRS status of the herds was evaluated based on the analysis of blood samples by another serological test (blocking ELISA) capable of differentiating between infection with PRRSV of the American type and European type. The specificity of the indirect ELISA test on meat juice samples was 0.98. The sensitivity of the test depended on the type of the PRRSV strain involved. The apparent prevalence in herds infected with the American type of PRRSV was 0.44. The apparent prevalence in herds infected with the European type of PRRSV was 0.64. Herd level sampling and herd level criteria for assessing the PRRS status of herds by the new test were developed. Herds were classified as PRRS negative or PRRS seropositive based on 10 meat juice samples collected randomly at slaughter throughout a 3-month-period. Herd PRRS status classification by the indirect ELISA was validated in 47 herds by collection of blood samples from the herds. Eighteen herds were classified as PRRS negative by both test systems. Twenty-nine herds were classified as PRRS seropositive by both test systems. Acceptable herd classification was achieved using this test.     

East coast fever as a cause of calf mortality in zanzibar

Crossbred and zebu calves at 4 to 5 months of age in an area where tick control was carried out showed little difference in mortality. Crossbred calves at 10 months of age had a mortality rate of 20% from all causes, 14% from East Coast fever. Serological results plus mortality studies showed that the survival rate of crossbred calves known to have been exposed to ECF was 57%. Serological results further showed that, in spite of tick exposure, only 26% of the crossbreds had seroconverted at 10.3 months of age while 15% zebus had positive titres.     

Studies on age specific leukocyte and lymphocyte counts in normal Friesian, Red Danish and Jersey cattle in Denmark.

To determine reference values for leukocyte and lymphocyte counts of the most common cattle breeds in Denmark, especially regarding the evaluation procedure in the leukosis control work, blood samples from normal cattle of all ages within the Friesian, the Red Danish and the Jersey breeds were collected, transported and counted as in the routine leukosis control programme. Evaluation of breed- and age specific counts showed numerically small, but statistically significant differences in mean values of leukocytes and lymphocytes between on the one hand Jersey cattle and on the other hand Red Danish and Friesian cattle. However, it was found that the key values used in the leukosis control work are generally in good agreement with the calculated 99.87 percentiles of all age- and breed specific lymphocyte count distributions, which were approximated with logarithmic normal distribution curves.     

Estimation of sensitivity, specificity and predictive values of two serologic tests for the detection of antibodies against Actinobacillus pleuropneumoniae serotype 2 in the absence of a reference test (gold standard)

Latent-class models were used to determine the sensitivity, specificity and predictive values of a polyclonal blocking enzyme-linked immunosorbent assay (ELISA) and a modified complement-fixation test (CFT) when there was no reference test. The tests were used for detection of antibodies against Actinobacillus pleuropneumoniae serotype 2 in a survey of respiratory diseases in Danish finishing pigs. The estimates were obtained by maximum-likelihood and also by a Bayesian method (implemented with Gibbs sampling). Possible dependence of diagnostic errors was investigated by comparing models where independence was assumed to models allowing for conditional dependence, given the true disease status.No strong evidence of conditional dependence in either test sensitivity or specificity was found. Assuming independence, maximum-likelihood estimates and 95% confidence intervals of the sensitivity and specificity of the ELISA were 100% and 92.8% (90.1–95.5%) and the corresponding values of the CFT were 90.6% (85.8–95.4%) and 98.6% (98.0–99.3%), respectively. Bayesian estimates and posterior 95% credible intervals of the sensitivity and specificity of the ELISA were 99.7% (98.7–100%) and 92.7% (89.9–95.3%) and of the CFT were 90.6% (86.0–95.3%) and 98.7% (98.0–99.3%). The sensitivity and specificity of a combined test, where the CFT is subsequently applied to the pig sera that test positive in the ELISA, were estimated at 90.2% (85.6–95.0%) and 99.9% (99.8–100%), respectively. The cost of the combined test was less than the cost of the use of the CFT alone, at prevalences <54%. Prevalences and predictive values and their 95% limits were estimated in six sub-samples of data. The estimates of sensitivity and specificity obtained in the present investigation generally validate those reported from other sources.     

Duration of load revisited

A duration of load study representing 13 years of testing was recently terminated. Preliminary results have been published over the years. This paper represents the final account of the study, which was focused on the influence of moisture content on time to failure for structural timber subjected to bending under constant load conditions. Two constant moisture conditions (MC = 11 and 20%) and one condition of varying moisture (MC between 11 and 20%) were applied. A total of 816 Norway spruce boards of dimensions 44 × 95 × 1,800 mm³ were included. Eight groups of non-destructively matched samples were formed. Four groups were subjected to short-term strength tests, and four groups were subjected to long-term tests. Creep and time to failure were monitored. Time to failure as a function of stress level was established and the reliability of stress level assessment was discussed. A significant mechanosorptive effect was demonstrated both in terms of increased creep and shortening of time to failure. The test results were employed for the calibration of four existing duration of load models. The effect of long-term loading was expressed as the stress level SL₅₀ to cause failure after 50 years of loading. SL₅₀ was found to be of the order 0.60 for MC = 11%, 0.50 for MC = 20% and 0.44 for MC varying between 11 and 20%. The test results revealed no evidence of a threshold stress level. A reliability based calibration of load-duration factors was performed using probabilistic models of loads and of the short-term and long-term strengths. For permanent and imposed library loads, reliability-based estimation of the load duration factor gave almost the same results as direct, deterministic calibration.     

Designing and evaluating risk-based surveillance systems: potential unwarranted effects of applying adjusted risk estimates

Risk-based surveillance systems reveal occurrence of disease or infection in a sample of population units, which are selected on the basis of risk factors for the condition under study. The purpose of such systems for supporting practical animal disease policy formulations and management decisions are: A: to detect an emerging disease or infection, if it becomes introduced into a population; or B: to substantiate freedom from a condition in a population; or C: to detect cases and estimate the prevalence of an endemic condition in a population. In risk-based surveillance these aims should be met with prudent use of resources while maintaining acceptable system performance. High-risk category units are selected for testing by identification of the presence of specific high-risk factor(s), while disregarding other factors that might also influence the risk. On this basis we argue that the most applicable risk estimate for use in designing and evaluating a risk-based surveillance system would be a crude (unadjusted) relative risk, odds ratio or apparent prevalence. Risk estimates found in the published literature, however, are often the results of multivariable analyses implicitly adjusting the estimates for confounding from other risk factors. We describe some potential unintentional effects when using adjusted risk estimates in evaluating the efficacy and sensitivity of risk-based surveillance systems (SSe). In two examples, we quantify and compare the efficacy and SSe using adjusted and crude risk estimates. The examples use Danish surveillance data from previously published studies to evaluate systems aimed at risk-based detection of new cases of an endemic infection, i.e. Salmonella in dairy cattle herds (Example 1), and for substantiating the absence of a specific infection, i.e. Trichinella in the national slaughter pig population (Example 2), respectively.