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The present paper aims to consider both problems in a more explicit way. After having given some information on the data used, we will review the variables which, on a priori grounds, are supposed to be of relevance as independent variables in our analysis. This set of variables is first analysed on expected linear relationships by means of a factor analysis. The results of this analysis are used to indicate the most important variables to be used in a regression-analysis, as to reduce the impact of multicollinearity. The residuals of the regression-analysis have been tested on the presence of spatial autocorrelation. The parameter-estimates obtained make it possible to draw some conclusions concerning the relative importance of the independent variables. [ABSTRACT FROM AUTHOR]

The objective of this cross-sectional study was to reevaluate the prevalence of digital dermatitis (DD) and associated risk factors in The Netherlands. Between May 2002 and December 2003, information about the presence or absence of DD lesions and other claw disorders on the hind claws of dairy cows in The Netherlands was collected by 20 hoof trimmers during trimming of all dairy cows in 383 herds. A questionnaire was used to acquire information regarding management and housing. Additional information, such as parity, breed, and stage of lactation of the cows, was provided by the Dutch Herd Book Organization. Digital dermatitis was present in 21.2% (SE = 0.3) of the study population (n = 22,454 cows). The herd prevalence varied from 0% (9.1% of the herds) to 83.0%. Most common was herd prevalence between 5 and 10%. The Holstein-Friesian breed was at higher risk for DD [odds ratio (OR) = 1.7] than was the Meuse Rhine IJssel breed (dual purpose breed). The risk for DD decreased with increasing parity. Cows at the peak of their lactation (30 to 60 d in milk) and in the third parity had higher odds for DD in comparison with cows that were later than 60 DIM. The presence of other claw disorders, such as interdigital dermatitis/ heel horn erosion (IDHE), interdigital hyperplasia (HYP), and interdigital phlegmon, appeared to be predisposing for DD. Based on estimation of the population-attributable fraction, it was concluded that if IDHE, HYP, and interdigital phlegmon were not present among the study population, respectively, 32.2, 9.0, and 1.1% of the DD cases could have been prevented. The risk for DD slightly decreased when cows affected by IDHE had access to pasture. Because of the interrelation between infectious claw disorders, an effective intervention strategy against DD should focus on an integrated approach to the control of all infectious claw diseases. Cows trimmed >12 me before the study (during regular trimming of the entire herd) were at lower risk for DD than were cows that were trimmed at shorter intervals. Animals that had >8 h of access to pasture were at higher risk for DD (OR = 1.6) compared with no access to pasture. Finally, cows in small-sized herds (<45 cows) affected with HYP were at lower risk (OR = 0.6) for DD than were cows affected with HYP in medium- and large-sized herds (60 to 85 cows per herd). [ABSTRACT FROM AUTHOR]

A study using an ELISA and a real-time PCR assay based on the detection of the repetitive transposon-like gene of Coxiella burnetii revealed that infection with the bacterium was widespread among Dutch dairy herds, with antibodies detected in bulk tank milk (BTM) from 268 of 341 herds (78.6 per cent) and bacterial DNA detected in 193 of 341 herds (56.6 per cent). The BTM samples were taken in November and December 2007. Serological and molecular studies in young and adult cattle selected from 100 herds showed that antibodies were present in the blood of 470 of 2936 (16.0 per cent) lactating cows but only in 19 of 1831 (1.0 per cent) young animals. Bacterial DNA was detected in the milk of 254 of 2925 (8.7 per cent) lactating cows; bacterial DNA was not detected in any of the faecal samples obtained from youngstock. The blood and milk samples were taken from the cattle in the period January to April 2008.

Bluetongue virus serotype 8 (BTV-8) emerged in the Netherlands in August 2006 for the first time. In the winter of 2006/2007, during the transmission free period, the government decided to establish a sentinel network to monitor the re-emergence of BTV-8 in 2007. Between June and December 2007, a sentinel network of 275 dairy herds with 8901 seronegative cows at start, was in place for BTV-8 testing in milk samples. Besides estimates of the monthly BTV-8 within-herd prevalence per region (south, central and north), this sentinel was used to determine BTV-8 associated risk factors. Information on management and housing practices that were hypothesized to be related to the increase in BTV-8 prevalence (risk factors) were used. Complete information on BTV-8 testing and management was obtained for 234 herds. At herd level, the increase in seroprevalence was defined as the total increase in seroprevalence amongst sentinel cows per herd during the sampling period (in months) divided by the number of sampling months in which the spread of BTV-8 occurred within the particular compartment in which the herd was located. This parameter was used as dependent variable in the linear regression analysis. The final model revealed four risk factors remained in the final model. Herds in the central and southern region of the Netherlands had a higher monthly increase in seroprevalence 6.4% (95% CI: 3.1-9.9) and 10.1% (95% CI: 6.2-14.3), respectively compared to herds in the northern region. Furthermore, there was a strong association with grazing. The monthly increase in seroprevalence in cattle pastured a few hours per day or throughout the day was 5.6% (95% CI: 1.4-10.2) to 11.4% (95% CI: 6.0-17.3) higher, relative to that for cattle kept indoors. For cattle that grazed outdoors throughout the day and the night, the monthly increase in seroprevalence was 13.6% (95% CI: 7.2-20.8). In addition, an association was found between the monthly increase in seroprevalence and some factors relating to stable design. Keeping the stable doors closed during the day was linked to a higher seroprevalence rate compared to that in stables with the door left open (3.6% (95% CI: 0.3-7.1)). Furthermore, a horizontal ventilation opening (>30 cm) along the walls of the stable, and with a windbreak curtain, appeared to offer some protection (-3.0% per month (95% CI: -6.0 to 0.2)) as compared to stables that had no or, only a small, ventilation opening (<30 cm). By the second half of 2007, bluetongue had spread over throughout the Netherlands. Our study indicated that there were some management factors that may help limit exposure to BTV-8 and its consequences., (Copyright 2009 Elsevier B.V. All rights reserved.)

A cross-sectional study was performed, to (re)evaluate the prevalence of sole ulcers (SUs) and the risk factors involved in dairy herds that perform claw trimming on a routine base. Twenty hoof trimmers collected data on the presence or absence of claw disorders on the hind claws of dairy cows during whole herd trimming in 383 herds in The Netherlands. A questionnaire was used to acquire information on management and housing. Additional data on parity, breed and stage of lactation of the cows were provided by the Dutch Herd Book Organization. Sole ulcer was present in 85% of the herds and in 5.6% (S.E. 0.2) of the study population (n=22,454 cows). The within-herd prevalence varied from 0% (14.6% of herds) to 26% (0.3% of herds) and in herds with SU was most frequently diagnosed 1-5% of the trimmed cows (45% of all herds). The risk for SU increased significantly with increasing parity. Cows that were at or after peak of lactation (>60 days in milk, DIM) had higher odds on SU in comparison to cows that were between 0 and 60 DIM (OR=0.6). The presence of other non-infectious claw disorders appeared to predispose to SU. Based on an estimation of the population attributable fraction, it was concluded that if sole hemorrhages, chronic laminitis (CL), white line disease and interdigital hyperplasia were the cause of SU and if those diseases could have been prevented in the study population, 35.4, 5.8, 11.6 and 4.6% of the SU cases respectively could have been prevented. Because of this relation, an effective intervention strategy of SU should focus on an integral approach of reducing the first stages of the bovine laminitis complex, whereas SU and CL are considered to be a later stage. Cows trimmed during late summer were at a higher risk for SU than cows trimmed in other periods of the year. Finally, cows in herds with purchased heifers were at a higher risk (OR=1.4) for SU than cows in herds that did not purchase heifers. A comparable positive effect was visible in herds with mattress bedding.

A serological follow-up study of 3.5 years duration was done of a dairy herd that had experienced a mass seroconversion to Neospora caninum following a point source exposure shortly before the 17th of January 2000. A total of 913 blood samples of 244 animals at seven sampling dates were used to investigate the seroprevalence dynamics in the herd. Most postnatally infected cattle remained seropositive during the period of investigation but 11 animals became seronegative after 6-27 months indicating transient infection. Six animals seroconverted later than the main group of 45 animals and 5 animals became seronegative after at least two seropositive records possibly due to a low infection dose or difference in the haplotypes of the infected animals. In total 58% (14/24) of the offspring of postnatally infected dams was seropositive. Nine of 16 (56%) daughters originating from inseminations after the postnatal infection of their dams were seropositive indicating endogenous transplacental infection.

Data of a cross-sectional study on prevalence of seven common bovine-claw disorders were used to calculate the intra-class correlation coefficients (ICCs) for claw trimmers. Fifteen professional claw trimmers were trained in diagnosing claw disorders uniformly. During regular trimming of all dairy cows in a herd, they recorded the presence (yes/no) of hind-claw disorders for digital dermatitis (DD), chronic laminitis (CL), sole ulcer (SU), white-line disease (WLD) and interdigital hyperplasia (HYP). For the claw disorders interdigital dermatitis/heel-horn erosion (IDHE) and sole haemorrhage (SH), claw trimmers also recorded (using graduated scores 1-3) the extent of the lesion. Complete information of 21,153 animals from 361 herds in The Netherlands was recorded between July 2002 and December 2003. To assure objective recording of claw disorders, ideally there would not be any correlation between observations. However, quantified ICCs from claw-trimmers observations ranged from 4.9% for DD to 38.0% for CL, while ICCs for herd ranged from 7.9% for SU to 26.7% for DD. Changes in the cut-off values for the diagnosis of IDHE and SH did not result in any relevant changes of the ICCs from claw-trimmers observations and these remained larger than the ICC explained by the herd. Based on these results, we conclude that for estimating measures of frequencies for different claw disorders, uniformity in recording certain diagnoses becomes an issue to consider when involving more than one observer.

Herd, within-herd and animal prevalences for Neospora caninum in beef and dairy cattle were compared between four countries. In randomly selected herds from regions of Germany, The Netherlands, Spain and Sweden that were representative for the cattle production of these countries, all animals > or = 2 years were examined serologically by enzyme-linked immunosorbent assays (ELISAs) with high test specificity (> 98.0%). In a previous study, the ELISAs had been validated against each other. Single reacting animals within a herd were confirmed by immunobloting. At the time of sampling, animal (age, breed, herdtype, sex, lactation stage) and herd data (region) were collected. Considerable differences in N. caninum herd, within-herd, and overall animal prevalence estimations were observed between countries, regions, herdtype, age categories and breeds. Herd prevalences, based on confirmation of single reactors, for dairy herds were estimated to be 16% (95%CI: 10-24%) in Sweden, 49% (95%CI: 39-59%) in Germany, 63% (95%CI: 57-69%) in Spain and 76% (95%CI: 67-84%) in The Netherlands and for beef herds 41% (95%CI: 31-50%) in Germany, 46% (95%CI: 41-51%) in Spain and 61% (95%CI: 50-72%) in The Netherlands. No beef herds were examined in Sweden. The lowest animal true prevalence was estimated in dairy cattle in Sweden (0.5% (95%CI: 0.1-0.8%)) while the highest animal true prevalence was estimated for dairy cattle in Spain (16.2% (95%CI: 14.9-17.5%)). Within-herd prevalences varied greatly, with very few farms in Sweden having more than 10% seropositive animals while in Spain more than 10% of the herds had within-herd prevalences between 50 and 100%. Seropositivity was significantly associated with herdtype (beef versus dairy), age, breed and region within countries. The results of this supranational comparative study showed that the importance of N. caninum infection varied greatly within in Europe. Estimates of prevalence can be used to calculate the economic impact of N. caninum infection as well as to evaluate the effect of prevention and control strategies over time.

Three ELISAs for the detection of antibodies against Neospora caninum in bulk milk were evaluated in 162 Dutch dairy herds. The first ELISA was the Dutch Animal Health Service (AHS) in-house ELISA, developed from the routine in-house serum ELISA. The other two ELISAs were commercial milk ELISAs from IDEXX and LSI. Blood samples of all lactating cows in 162 dairy herds were tested using the AHS in-house serum ELISA. Based on previous studies in the Netherlands a within-herd N. caninum seroprevalence of 15% was associated with increased risk for reproductive losses. This percentage was therefore used as positive seroprevalence cut-off value. Repeatability of the ELISAs was evaluated by testing on three different days. The AHS in-house ELISA lacked specificity, probably due to use of a different batch of antigen on the second and third test-day. Cut-off values were determined using misclassification costs term calculations. At cut-off values 0.6 for the IDEXX and 0.2 for the LSI, a herd sensitivity of 61% (95% CI: 49--73%) and 47% (95% CI: 35--60%) was estimated. Herd specificity at these cut-off values was 92% (95% CI: 87--98%) for the IDEXX and 94% (95% CI: 90--99%) for the LSI ELISA. The positive and negative predictive values were 84% (95% CI: 68--100%) and 86% (95% CI: 79--94%) for the IDEXX ELISA, and 85% (95% CI: 67--100%) and 82% (95% CI: 74--90%) for the LSI ELISA. The agreement between all possible combinations of test-days was expressed by kappa values. These were found to be slightly higher for the IDEXX than for the LSI ELISA. It is concluded that both commercial ELISAs performed satisfactorily to detect a within-herd seroprevalence of N. caninum in lactating cows of at least 15%.

The optimal moment for trimming the claws of all dairy cows in a herd was investigated by assessing the external rotation of the hind claws of individual cows relative to the spinal column. This leg score consisted of three independent descriptors: 1 (good/normal), 2 (moderately deviant), and 3 (severely deviant). This study assessed the repeatability and the reproducibility of the leg score system, and the consistency of the advice given subsequently about trimming of the hind claws of all cows in the herd. Repeatability was assessed for 52 cows that were scored twice on the same day by 11 observers; the kappa value ranged from 0.17 to 0.66 (mean: 0.36). The probability of the same result for both assessments ranged from 0.49 to 0.80 (mean: 0.61). Claw trimming was advised if at least 20% of the cows had a leg score of 3. On the basis of the scores, 3 observers consistently advised trimming of the hind claws of all the cows in the herd, and 6 observers consistently advised against the need for trimming in the short term; 2 observers had an inconsistent advice. The reproducibility of the scoring system was assessed in two dairy herds (62 and 50 cows). Eight observers evaluated the leg score of the cows of both herds on the same day. The mean kappa value of the leg score for all pairs of different observers (A-B, A-C etcetera) was 0.24 [-0.08-0.49]. In conclusion, the leg score is not a reliable method for determining the optimal moment for claw trimming in dairy cattle. The reasons for the inconsistent observations require further investigation.

An outbreak of foot-and-mouth disease (FMD) in Great Britain was reported on 21 February 2001, followed by an outbreak of FMD in The Netherlands a month later. This Dutch index outbreak occurred on a mixed, veal-calf/dairy-goat farm in Oene, in the central part of The Netherlands. The most-likely route of infection was the import of Irish veal-calves to this Dutch herd via an FMD-contaminated staging point in France. With hindsight, more herds seemed to be infected by the time the index outbreak was confirmed. The regular EU control measures were implemented, in combination with pre-emptive culling of herds within 1km of each outbreak. Nevertheless, more outbreaks of FMD occurred. Most of the virus infections on those farms were "neighborhood infections". Because the situation seemed out of control locally and the destruction capacity became insufficient, it was decided to implement an emergency vaccination strategy for all biungulates in a large area around Oene to stop further spread of the virus. All susceptible animals on approximately 1800 farms in this area were vaccinated. All farms subsequently were depopulated, starting from 2 weeks after vaccination. In total, 26 outbreaks were detected (the last outbreak on 22 April 2001). In total, approximately 260,000 animals were killed.

Risk factors for outbreaks in 1999 of clinical Salmonella enterica subsp. enterica serovar Typhimurium infection on dairy farms were studied in a matched case-control study with 47 case farms and 47 control farms. All 47 case farms experienced a clinical outbreak of salmonellosis which was confirmed with a positive bacteriologic culture for serovar Typhimurium in one or more samples. Serovar Typhimurium phage type 401 and 506 (definitive type 104, DT104) were the most frequently isolated phage types (13 isolates). On most farms (66%), clinical signs were seen only among adult cows. The most frequently reported clinical signs were diarrhoea (in 92% of the farms) and depression (in 79% of the farms). Control farms were matched on region and had no history of salmonellosis. A questionnaire was used to collect data on case and control farms. The relationship between serovar Typhimurium status of the farm and possible risk factors was tested using conditional logistic regression. Significant factors in the final model were presence of cats on the farm (OR=0.06), purchase of manure (OR=21.5), feeding colostrum only from own dam (OR=0.08), a non-seasonal calving pattern (OR=25), unrestricted grazing of lactating cows (OR=0.07), and a high mean mowing percentage of pasture (OR=1.02).

In 517 Dutch families at a family cancer clinic, we screened for BRCA1/2 alterations using the Protein Truncation Test (PTT) covering approximately 60% of the coding sequences of both genes and direct testing for a number of previously identified Dutch recurrent mutations. In 119 (23%) of the 517 families, we detected a mutation in BRCA1 (n=98; 19%) or BRCA2 (n=21; 4%). BRCA1/2 mutations were found in 72 (52%) of 138 families with breast and ovarian cancer (HBOC), in 43 (13%) of the 339 families with breast cancer only (HBC), in 4 (36%) of 11 families with ovarian cancer only (HOC), and in nine of 29 families with one single young case (<40 years) of breast cancer. Between the different subgroups of families (subdivided by the number of patients, cancer phenotype and age of onset) the proportion of BRCA1/2 mutations detected, varied between 6 and 82%. Eight different mutations, each encountered in at least six distinct families, represented as much as 61% (73/119 families) of all mutations found. The original birthplaces of the ancestors of carriers of these eight recurrent mutations were traced. To estimate the relative contribution of two important regional recurrent mutations (BRCA1 founder mutation IVS12-1643del3835 and BRCA2 founder mutation 5579insA) to the overall occurrence of breast cancer, we performed a population-based study in two specific small regions. The two region-specific BRCA1 and BRCA2 founder mutations were detected in 2.8% (3/106) and 3.2% (3/93) of the unselected breast tumours, respectively. Of tumours diagnosed before the age of 50 years, 6.9% (3/43) and 6.6% (2/30) carried the region-specific founder mutation. Thus, large regional differences exist in the prevalence of certain specific BRCA1/BRCA2 founder mutations, even in very small areas concerning populations of approximately 200000 inhabitants.

On 23 February 1999, the Dutch Animal Health Service advised all Dutch veterinary practices to postpone vaccination against bovine herpesvirus 1 (BHV1) immediately. The day before severe disease problems were diagnosed on four dairy farms after vaccination with the same batch of BHV1 marker vaccine. Using monoclonal antibodies, bovine virus diarrhoea virus (BVDV) type 2 was found in the vaccine batch. This paper describes an outbreak of BVDV type 2 infection caused by the use of a batch of modified live BHV1 marker vaccine contaminated with BDVD. Sources of information used were reports of farm visits, minutes of meetings, laboratory results, and oral communications from the people involved. The first symptoms of disease were observed on average six days after vaccination. Morbidity was high on 11 of the 12 farms. On five farms more than 70% of the animals became ill, while on one farm no symptoms could be detected. During the first week after vaccination, feed intake and milk production decreased. During the second week, some animals became clinically diseased having nasal discharge, fever, and diarrhoea. At the end of the second week and at the start of the third week, the number of diseased animals increased rapidly, the symptoms became more severe, and some animals died. Mortality varied among herds. Necropsy most often revealed erosions and ulcers of the mucosa of the digestive tract. In addition, degeneration of the liver, hyperaemia of the abomasum, and swollen mesenterial lymph nodes and swollen spleen were found. On 11 of the 12 farms all animals were culled between 32 and 68 days after vaccination after an agreement was reached with the manufacturer of the vaccine. This was the third outbreak of BVD in cattle after administration of a contaminated vaccine in the Netherlands. The possibilities to prevent contamination of a vaccine as a consequence of infection of fetal calf serum with BVDV are discussed. Improvement of controls to prevent contamination before and during vaccine production, and improvement of the monitoring of side-effects is necessary.

Neospora caninum is an intracellular protozoan parasite that was discovered in a dog in 1988. Since then, N. caninum has been demonstrated in a variety of animal species and it has been recognized as an important cause of abortion in cattle. An infection with N. caninum can be maintained in cattle herds for several generations by transplacental transmission from cow to calf. Recently, it was demonstrated that dogs can act as definitive hosts of N. caninum and therefore may be a source of infection for other species by shedding oocysts. Further evidence of a role of the dog in spreading the infection to cattle has been derived from epidemiological studies. The present state of knowledge is reviewed in this paper.

Abortion storms in 50 dairy herds in The Netherlands were reported in which there was a strong association with Neospora caninum-infection. The duration of the abortion storms ranged from 6 to 65 d (mean 41.5 d). The cumulative proportion of aborting cows ranged from 0.11 to 0.57 (mean 0.26) of the animals at risk. An apparent seasonal influence was noted as most abortion storms occurred during the summer and early autumn. The prevalence of antibodies to N. caninum in 50 herds which had had an abortion storm was compared with that of 100 control herds which had no history of an abortion storm. Seroprevalence was estimated by testing a 20% cross sectional herd sample using a tachyzoite lysate-based ELISA method. Seroprevalence in case herds (range 17 to 87%, mean 51.5%) was significantly higher than that in control herds (range 0 to 53%, mean 13.9%). For most herds the seroprevalence levels were equal across all age groups, which suggests that the infection had been perpetuated by vertical transmission. In these herds, the abortion storms appeared to be induced by factors causing recrudescence of a N. caninum-infection in chronically infected animals rather than being the result of a recent introduction. In 6 case herds the seroprevalence in the dairy cows was significantly higher than in the young stock, which may have been attributable to superimposed postnatal infection.

A 2 to 1 matched case control study design was used to analyze herd level risk factors for Neospora caninum-associated abortion storms in 47 dairy herds. Data were obtained using a questionnaire regarding the state of affairs at the farms over the 2 years prior to the abortion storm. The questionnaire included 120 variables considered to be potential risk factors for either introduction of infection or recrudescence of chronic infection. The relationship between risk factors and case control pairs was analyzed by conditional logistic regression using a three-steps procedure. In addition, cross sectional serology was used to assess the possible role of concomitant infections. The main factors that were significant in the analysis and that were considered to have potential biological relevance were the presence of dogs, the presence of poultry, and the feeding of moldy maize-silage during summer. For both the presence of dogs and the presence of poultry on the farms, a linear relationship was found between the number of animals and the assessed risk for an abortion storm. These findings suggest a possible role of these species in the transmission of N. caninum. Further evidence for such a role of dogs was the significant association between the presence of dogs and the presence of seropositive cattle in the control herds. The feeding of moldy fodder is considered to be a factor which may induce recrudescence of a latent N. caninum-infection by mycotoxins causing immune suppression. We also found some evidence for a possible influence of management practices around calving and a high prevalence of retained afterbirths. No significant association was found for herd level prevalence of antibodies to bovine viral diarrhea virus, bovine herpesvirus 1, Leptospira hardjo or Salmonella dublin.

Laboratory results obtained in different laboratories over lengthy periods of time usually are difficult to compare. In cooperative long-term studies where such results must be pooled, thorough standardization of methods is vital. We describe a program in which comparable plasma cholesterol and glucose analyses have been obtained, by simple methods. In the Netherlands and the Soviet Union in close collaboration with the Center for Disease Control, Atlanta, Ga., U.S.A. The two laboratories produced glucose values (direct o-toluidine reaction) within 2% of the target reference values and cholesterol results (direct Liebermann-Burchard reaction) with a consistent 6-8% positive bias over the reference method values. Intralaboratory precision was subject to preset acceptance limits. The use of common control materials, exchange of patient samples, and on-site comparison of all details of laboratory procedures are vital tools in standardization efforts. A laboratory protocol that included quality requirements and rejection criteria was developed and proved to be indispensable. The experience gained should be useful in standardizing inter-laboratory results in similar studies.