Clinical and laboratory evaluation of a real-time PCR for Clostridium difficile toxin A and B genes.

Many laboratories use enzyme immunoassays (EIAs) for the diagnosis of Clostridium difficile infection (CDI). More recently, polymerase chain reaction (PCR)-based diagnosis has been described as a sensitive test. Real-time PCR for the detection of C. difficile toxin A and B genes was evaluated. A prospective evaluation was performed on stool samples from 150 hospitalized adult patients and 141 healthy volunteers. PCR was compared to toxigenic culture (TC), direct cytotoxicity test (CTT), ImmunoCard® Toxin A and B (Meridian Bioscience), and enzyme-linked immunosorbent assay (ELISA) (Vidas). The results were correlated with clinical data using a standardized questionnaire. The diagnostic yield of the PCR was further evaluated after implementation. Using toxigenic culture as the gold standard, the sensitivity and specificity of PCR were 100 and 99.2%, respectively. Patients were categorized as follows: TC/PCR-positive (n = 17) and negative TC (n = 133). The differences in these groups were more frequent use of antibiotics and leukocytosis (p < 0.05). The diagnostic yield of PCR was evaluated during a period of 6 months and showed an increase of positive patients by 50%. PCR for the detection of toxigenic C. difficile has a high sensitivity and can rule out CDI, but cannot differentiate CDI from asymptomatic carriage. Clinicians should be aware of this in order to prevent inappropriate treatment and delay of other diagnostics.

Performance of two tube coagulase methods for rapid identification of Staphylococcus aureus from blood cultures and their impact on antimicrobial management.

Test parameters and clinical impact of the direct tube coagulase test (DTCT) for rapid identification of Staphylococcus aureus from blood culture were investigated. The sensitivity of the DTCT at 4 h using saline dilution was 96%, compared with 93% using serum separator tubes; specificity was 100% for both methods. Among 32 patients with S. aureus bacteraemia, treatment modifications were based on microbiology results from the primary source of infection in 12 patients, on a Gram's stain from blood culture in seven patients, and on the DTCT in nine patients. The DTCT is a valuable adjunct in the routine microbiology laboratory because of its good performance, technical simplicity and low cost.

Natural postnatal Neospora caninum infection in cattle can persist and lead to endogenous transplacental infection.

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.

Herd and cow-level prevalence of sole ulcers in The Netherlands and associated-risk factors.

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.

Supranational comparison of Neospora caninum seroprevalences in cattle in Germany, The Netherlands, Spain and Sweden.

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.

Intra-class correlation attributable to claw trimmers scoring common hind-claw disorders in Dutch dairy herds.

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- and cow-level prevalence of digital dermatitis in the Netherlands and associated risk factors.

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 mo 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).

Overexpression of the insulin-like growth factor-1 receptor and autocrine stimulation in human cervical cancer cells.

We characterized mechanisms of growth control involving insulin-like growth factor-1 (IGF-1), IGF-2, and IGF-1 receptor (IGF-1R) by investigating their expression in human cervical cancer cell lines, primary cervical tumor cell cultures, and normal ectocervical epithelial cells maintained in short-term culture. By reverse transcription followed by PCR, IGF-1 mRNA was not detected in any of the cell lines, whereas IGF-2 mRNA transcripts were detected in all of them. Using the RNase protection assay, low levels of IGF-2 mRNA were also detected in all of the cervical cancer cell lines, primary cervical tumor cell cultures, and normal ectocervical cultures tested, but no IGF-1 transcripts were detected. Scatchard analysis revealed 3- and 5-fold increases in IGF-1R expression by the primary cervical cancer cell cultures and cervical cancer cell lines, respectively, compared with the normal ectocervical cells. In proliferation assays, epidermal growth factor (EGF) consistently enhanced cervical cancer cell growth, but an antisense oligonucleotide to IGF-2 uniformly inhibited the EGF-induced mitogenic effect. These studies suggest that autocrine production of IGF-2 and overexpression of the IGF-1R are important components controlling the proliferation of cervical carcinoma cells, and that autocrine IGF-2 production in cervical cancer cells may participate in the mitogenic signaling of EGF.

Elevated pentose cycle and glucuronyltransferase in daunorubicin-resistant P388 cells.

Anthracycline resistance of P388 daunorubicin-resistant cells cannot be accounted for merely by differences in drug uptake and retention; protection against intracellular drug was also indicated. Cytotoxicity of daunorubicin may be partially due to the formation of free radicals and reactive oxygen species (hydrogen peroxide, hydroxyl radical, singlet oxygen, and superoxide anion radical). Protection against free radicals and peroxides is largely dependent upon the availability of reduced glutathione, which in turn requires NADPH for its continual regeneration. Pentose phosphate cycle (also called hexose monophosphate shunt) is known to provide NADPH for maintenance of glutathione. Activities of the two NADPH-producing dehydrogenases of the cycle, glucose-6-phosphate and 6-phosphogluconate dehydrogenase, were 40% higher (P less than 0.05) and activity of the cycle in intact cells was 2-fold higher in the resistant than the sensitive cells. The cycle was as active in these cells as it is known to be in macrophages, indicating a very effective protection against oxidative stress, free radicals, and alkylating electrophiles. Elevated activity of the pentose phosphate pathway in drug-resistant cells can represent a mechanism of resistance against multiple structurally unrelated drugs. Efflux of daunorubicin may be aided by further metabolism to glucuronides. Daunorubicinol, a known active metabolite of daunorubicin, can be metabolized to a glucuronide by the cells and eliminated into the surrounding medium. Glucuronidation of daunorubicinol was evidenced by (a) release of daunorubicinol following glucuronidase hydrolysis of media from cell incubations with 1.8 microM daunorubicin and (b) production of radioactive glucuronide when cell homogenates were incubated with UDP-[14C]glucuronic acid plus daunorubicinol. Glucuronyltransferase activity with a broad substrate specificity was found in the cells. Using model substrates, 1-naphthol and o-aminophenol, it was determined that glucuronyltransferase activity was 4 times higher in daunorubicin-resistant than -sensitive P388 cells. Elevated glucuronyltransferase could contribute to daunorubicin and multidrug resistance.

Evaluation of three enzyme-linked immunosorbent assays for detection of antibodies to Neospora caninum in bulk milk.

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%.

Assessing the repeatability and reproducibility of the Leg Score: a Dutch Claw Health Scoring System for dairy cattle.

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.

Risk factors for infection with Foot-and-Mouth Disease virus in a cattle population vaccinated with a non-purified vaccine in Iran.

In this study, we estimated the level of Foot-and-Mouth (FMD) virus infection in a cattle-dense north-western province of Islamic Republic of Iran and analyzed putative risk factors for FMD infection. Calves (6-24 months of age) from all 17 districts of West Azerbaijan were tested for antibodies against non-structural proteins (NSP-Ab) of FMD virus. A proportional stratification with a minimum of 30 epi-units was applied for 3 different husbandry systems: villages, dairy and mixed farms. Within an epi-unit, 30 calves were sampled. For the interpretation of ELISA test results, we used the 50% inhibition (50PI) cut-off as per producer's instructions and created one at 75% inhibition (75PI) based on the lowest point of the histogram of PI results. This approach resulted in three categories of outcomes; negative (N), low-positive (LP) and high-positive (HP). A generalized mixed-effect model for binary outcomes was used for analysing putative risk factors and was run for both cut-off values. A total of 8378 calves from 202 villages, 51 dairy farms and 28 mixed farms were eligible for analysis. The percentage of calves testing positive (LP+HP) was 53.7% (95% Confidence interval (CI): 52.6%-54.8%), with 39.6% (95% CI: 38.6-40.7%) testing HP (n=3309) while 14.1% (95% CI: 13.5-15.0%) of calves tested LP (n=1188). Of 281 epi-units sampled, all calves sampled tested negative in only 2 epi-units (0.7% (95% CI: 0.1-2.5%)) and more than 25 calves tested positive in 29 epi-units (10.3% (95% CI: 7.0-14.5%)). Outcomes of regression modelling using the 50 PI cut-off indicated that, for each month increase in age, the odds of testing positive increased 1.01 times (95% CI: 1.00-1.03). The odds of calves testing NSP-positive increased 1.46 times (95% CI: 1.22-1.77) for calves residing in epi-units that had experienced clinical FMD in the 12 months preceding this study. The odds of calves owned by livestock owners who traded livestock testing positive were 1.4-1.6 times higher than those owned by persons not engaged in trading while the odds for calves testing positive in dairy herds was 1.62 (95% CI: 1.10-2.35) times higher compared with calves in villages. The results of the model using the 75 PI cut-off value resulted in comparable estimates, with the age-effect becoming more evident. These results have confirmed widespread FMD infection and were used in developing a risk-based control strategy on FMD, in line with Stage 1 of the Progressive Control Pathway for FMD (PCP-FMD).

Disturbances of cerebrospinal fluid flow attributable to arachnoid scarring cause interstitial edema of the cat spinal cord.

OBJECTIVE: Spinal arachnoid scarring may be caused by trauma, inflammation, surgery, spinal instability, degenerative diseases, or malformations and may lead to progressive neurological deficits and syringomyelia. We wanted to investigate the effects of focal arachnoid scarring in the cervical spinal canal of cats on pressures in the subarachnoid space and spinal cord tissue, as well as on spinal cord histological features. METHODS: Twenty-nine adult cats were used for this study. Nine animals served as control animals, whereas 20 animals received a focal arachnoid scar at C1-C2, which was produced by placement of a kaolin-soaked fibrin sponge on the posterior surface of the spinal cord. After 4 months, pressure recordings above and below the scar, in the subarachnoid space and spinal cord, were performed. Elasticity measurements were performed with small bolus injections. Morphometric analyses of brain and ventricle volumes, sizes of the central canal, and sizes of the perivascular spaces in gray and white matter were also performed. RESULTS: No animal developed clinical or neurophysiological evidence of neurological symptoms at any time. In the kaolin-treated group, pressure recordings revealed a significant increase in the subarachnoid pressure at C1, because of the cerebrospinal fluid flow obstruction. Pressure gradients tended to increase at all measuring points. A significant difference was detected between the spinal cord and subarachnoid space at C2, where the intramedullary pressure exceeded the subarachnoid pressure. Elasticity was significantly increased in the spinal cord at C2. Intracranially, no evidence of hydrocephalus was observed. In the spinal cord, perivascular spaces were significantly enlarged in the posterior white matter above the arachnoid scar and in the central gray matter below the area of scarring in the cervical cord. CONCLUSION: Arachnoid scarring at C1-C2 produces an interstitial type of edema in the central gray matter below the area of scarring in the cat cervical cord, because of altered cerebrospinal fluid and extracellular fluid flow dynamics. These changes may be interpreted as the initial stage in the development of syringomyelic cavities.

Characteristics of Neospora caninum-associated abortion storms in diary herds in The Netherlands (1995 to 1997).

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.

Risk factors for Neospora caninum-associated abortion storms in dairy herds in The Netherlands (1995 to 1997).

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.

[The role of the dog in the epidemiology of neosporosis in cattle]. / De rol van de hond in de epidemiologie van neosporose bij het rund.

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.

[Comparison of performance of dairy herds that were or were not vaccinated with a bovine herpes virus 1 marker vaccine in 1998]. / Vergelijking van de bedrijfskengetallen van melkveebedrijven die in 1998 wel of niet hebben gevaccineerd met BHV1 markervaccin.

This study analysed the effects of the use of bovine herpesvirus 1 (BHV1) marker vaccine on the performance of dairy cattle. In Spring of 1999, vaccination of 12 herds with the BHV1 marker vaccine resulted in severe animal health problems and mortality. The vaccines used on these farms were all from a batch that appeared to be contaminated with bovine virus diarrhoea virus type 2. This led to a general call to farmers and veterinary practitioners to report side-effects of this vaccine. As a result, more than 7000 farmers reported symptoms. The information was obtained by means of a questionnaire; there was no control group. To determine the effects of the use of the marker vaccine, it was necessary to perform a study based on objectively acquired information. The information collected by the Royal Dutch Cattle Syndicate and the office of Identification and Registration was complied into herd indices on production, udder health, reproduction, and culling. Two groups of dairy farms that had used the BHV1 marker vaccine (attenuated and inactivated vaccine) were compared with farms that were certified BHV1-free. The analyses were performed based on intra-herd comparisons, meaning that per herd each index calculated over a certain period of time after the use of the marker vaccine was compared to a similar period of time prior to the use of the marker vaccine. A total of 144 comparisons were made. Seven comparisons were statistically significant. In two comparisons, the results were in favour of the BHV1-free farms and in five comparisons, the result were in favour of the vaccinated farms. Thus use of the BHV1 marker vaccine could not be proven to affect herd performance. The sensitivity of the tests was very high, so with a high level of probability even very small differences in indices between groups would have been detected.

[Management and herd performance of dairy herds with and without chronic wasting disease]. / Bedrijfsvoering en kengetallen van tot slijterbedrijven verworden melkveebedrijven.

'Chronic wasting' in cattle acquired a special meaning in the Netherlands in 1999. It was used to define animal health problems that were thought to be associated with the use of bovine herpesvirus 1 marker vaccine. Criteria have not been set by which an objectively independent inventory of the problems could be made. The objective of this study was to determine management factors associated with the problem of 'chronic wasting' prior to the use of the BHV1 marker vaccine. Knowledge about these factors could be helpful for generating additional hypotheses about the aetiology of chronic wasting in cattle. A total of 188 farms participated in the study, of which 94 had severe problems with chronic wasting. The other half consisted of control farms matched with the case farms that did not report problems after the use of the BHV1 marker vaccine. Data analyses were performed over the period before (and not at the time of) 'chronic wasting' problems. Data were collected from various sources. A questionnaire was used to collect information on farm management practice. In addition, information on laboratory submissions for 1996 to 1998, animal movements in 1998, roughage analyses of 1997 and 1998, expenses for animal health in 1998, and herd performance in 1995 to 1999 was collected. In the analyses, a distinction was made between information obtained objectively and subjectively. Herds with problems of 'chronic wasting' were larger than herds without wasting problems (animals, surface) but not more intensively managed. 'Wasting' herds had a lower performance in terms of fertility and udder health. In addition, these herds had more contact with other herds through the purchase of animals. There were no differences in farm management practices related to disease control and prevention. Additional studies are required with regard to the patho-physiology of chronic wasting cows. The role of herd size needs more study.

[Vitamin B12 supplementation and milk production on farms with 'chronic wasting' cattle]. / Vitamine B12-suppletie en melkproductie op bedrijven met 'slijtende' runderen.

From early 1999 onwards, cattle health problems accompanied by chronic wasting of unknown aetiology were reported on a number of dairy farms. An association between these health problems and the compulsory use of gE-negative marker vaccines against bovine herpesvirus 1 was presumed by farmers. On one dairy farm an increased milk production of 50% was reported within a few days after parenteral vitamin B12 treatment. Therefore, the current study was designed to determine the effect of parenteral vitamin B12 treatment on the milk production of dairy herds with wasting cattle. A randomized blind trial was performed in five problem herds and two control herds. On each farm five lactating cows were injected intramuscularly with 20 mg vitamin B12 and paired with five untreated lactating cows. The milk production of treated and untreated animals was measured for 19 days following treatment and compared to pre-treatment production. No effect of vitamin B12 treatment on milk production was established on either problem farms or control farms. Neither was a difference detected in the response to vitamin B12 treatment between problem herds and control herds. In a second experiment, parenteral vitamin B12 treatment was applied in three problem herds by local veterinary practitioners. The results of this experiment were in line with the results of the first experiment.