Ovine and Bovine Congenital Abnormalities Associated With Intrauterine Infection With Schmallenberg Virus.

In December 2011, a previously unknown congenital syndrome of arthrogryposis and hydranencephaly in sheep and cattle appeared in the Netherlands as an emerging epizootic due to Schmallenberg virus (SBV). Gross lesions in 102 lambs and 204 calves included porencephaly, hydranencephaly, cerebellar dysplasia and dysplasia of the brainstem and spinal cord, a flattened skull with brachygnathia inferior, arthrogryposis, and vertebral column malformations. Microscopic lesions in the central nervous system showed rarefaction and cavitation in the white matter, as well as degeneration, necrosis, and loss of neurons in the gray matter. Brain and spinal cord lesions were more severe in lambs than in calves. Ovine and bovine cases examined early in the outbreak showed encephalomyelitis. SBV infection was confirmed by real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) in brain samples in 46 of 102 lambs (45%) and in 32 of 204 calves (16%). Immunohistochemistry, performed on tissue samples from 18 RT-qPCR-positive lambs, confirmed the presence of bunyaviral antigen in neurons of the brain in 16 cases. SBV antibodies were detected by enzyme-linked immunosorbent assay in fetal blood in 56 of 61 sampled ovine cases (92%). In a virus neutralization test, all tested dams of affected newborns, 46 ewes and 190 cows, were seropositive. Compared with other teratogenic viral infections, the pathogenesis and lesions of SBV in sheep and cattle fetuses are similar to those of other ruminant orthobunyaviruses. However, the loss of spinal ventral motor neurons and their tracts, resulting in micromyelia, distinguishes SBV infection from other viral central nervous system lesions in newborn ruminants.

[Postvaccinal fatal Streptococcus zooepidemicus necrotizing fasciitis in a young dog: a case report]. / Fatale necrotiserende fasciitis door Strepto- coccus zooepidemicus na vaccinatie bij een jonge hond, een case report.

A 2.5-years-old female mongrel dog was routinely subcutaneously vaccinated. A few hours later mental dullness was noticed by the owner progressing into stupor the next day and resulting in a comatose state and death within 48 hours after vaccination. At post mortem examination, which was extended with histology and bacteriology, a necrotizing fasciitis and bacteremia caused by Streptococcus equi subsp. zooepidemicus were established. In the isolated Streptococcus strain four different superantigens were demonstrated that appeared to be able to produce exotoxins in vitro. Therefore, it is concluded that the minor skin trauma caused by vaccination enabled this strain to gain access to the subcutaneous tissue and to induce a necrotizing fasciitis. This process was complicated with a bacterial septicemia leading to death of the dog within 48 hours.

Severe combined immunodeficiency in Frisian Water Dogs caused by a RAG1 mutation.

Mortality of pups at 8-12 weeks of age was frequently observed in Frisian Water Dogs. Blood parameters and clinical signs of newborns from three litters were monitored. Three pups from two litters showed strongly reduced levels of immunoglobulins and lymphocytes. These dogs were euthanized after first display of disease. Concurrent clinical and pathological features were consistent with a diagnosis of severe combined immunodeficiency (SCID). Defective V(D)J recombination is one of the causes of SCID in humans and animals. Eight genes involved in V(D)J recombination were investigated by segregation analysis of closely located microsatellite markers and by DNA sequence analysis. A nonsense mutation in the gene coding for V(D)J recombination factor RAG1 was identified in DNA from the cases at a position similar to that of nonsense mutations found in human SCID. It was concluded that SCID due to a mutation of RAG1 led to the high mortality.

[Claw health in Dutch dairy herds, an update of some recent disorders]. / Klauwgezondheid in Nederland, een update van enkele recente ziektebeelden.

The dairy practitioner can make an important contribution to animal welfare by preventing and treating painful claw disorders. Two relatively new claw disorders have been observed in Dutch dairy herds, namely, toe ulceration and non-healing white line disorder. This article provides an update of knowledge on these two disorders, the aetiology of which is not known yet.

[Botulism in dairy cattle in 2008: symptoms, diagnosis, pathogenesis, therapy, and prevention]. / Botulisme bij melkkoeien anno 2008: symptomatologie, diagnostiek, pathogenese, therapie en preventie aan de hand van een catastrofaal verlopen casus.

Botulism affects about 20 dairy herds a year in the Netherlands. This article describes the dramatic outcome of botulism in a dairy herd. The main clinical symptoms in this herd were increased lying down, slight ataxia of the hind legs, and a high mortality (98%). The diagnosis is difficult to establish in adult cattle, and for this reason the clinical and laboratory findings, differential diagnosis, therapy, and preventive measures are discussed. On the basis of this outbreak, previous experience with botulism, and cases described in literature, it is suggested that presence of 'free-range" poultry could contaminate grazing pastures with botulism neurotoxins, causing clinical problems in cattle. If there is an increased risk of contamination of the pasture and/or silage with botulinum neurotoxins, vaccination should be considered to prevent substantial economic and emotional damage.

Comparison of clinical and pathological diagnoses in dogs.

Clinical and pathological diagnoses were compared in a prospective study of 145 dogs. A diagnostic work up had been performed on all dogs of which 36 (24.8%) died and 109 (75.2%) were euthanatized. In 119 dogs (82.1%) both a clinical and patholical diagnosis was made, in 20 dogs (13.8%) no pathological diagnosis could be made and in 6 dogs (4.1%) no clinical diagnosis was established. In the 119 dogs the agreement level between clinical and pathological diagnosis was scored by the referring veterinarian together with a pathologist. Total agreement was found in 61 cases (51.3%) and disagreement in 31 cases (26.0%). In the remaining cases (27=22.7%) the pathological diagnosis further specified the clinical diagnosis. Consecutive submission appeared difficult to achieve by the participating veterinarians. However, no major differences in agreement level was present between the veterinarian which succeeded in almost consecutive submissions and the other veterinarians. At necropsy 42 cases were diagnosed as neoplasia, of which 52.4% had been diagnosed clinically. As to infectious diseases 55.0% of these diseases diagnosed at necropsy had been diagnosed clinically. In about 20% of the cases the differences were of clinical significance according to the referring veterinarians. In addition, it was indicated by the clinicians that about 50% of the necropsies revealed findings which could amend future patient care. The results of the study stress the relevance of postmortem examination as crucial part of continuing education and of quality monitoring and assurance in veterinary medicine.

Thyrotrophin receptors in normal and neoplastic (primary and metastatic) canine thyroid tissue.

Thyrotrophin (TSH) is the conditional growth factor of thyroid epithelial cells. Abnormalities in TSH-receptor binding such as a low receptor number or low binding affinity may be a marker of thyroid carcinoma or metastases, or may exhibit a relationship with the functional variability of such tissues. The dog was used as a model to characterize TSH-receptor binding in normal thyroid tissues, naturally occurring thyroid neoplasms and distant metastases. In normal dog thyroid tissues, specific 125I-labelled TSH binding ranged from 2.7 to 15.5%, and low cross-reactivity with bovine LH (0.023%) was observed. One class of TSH-binding sites was found in eight normal thyroid tissues and 22 thyroid carcinomas; two normal thyroid tissues and one tumour exhibited two classes of binding sites. The concentration of binding sites was lower in the five carcinomas with reduced pertechnetate uptake (0.09 pmol/mg protein) than in the five thyroid neoplasms with increased uptake (0.19 pmol/mg) (P = 0.055). Compared with the original carcinoma tissues, TSH binding revealed a reduced binding affinity in eight out of eleven metastases. Two metastases showed a complete absence of TSH binding, suggesting that they were not dependent on TSH for growth. We conclude that one class of TSH-binding site is predominant in normal dog thyroid tissues and dog thyroid carcinomas. TSH could therefore contribute, at least in theory, to further growth of primary dog thyroid carcinomas. Secondly, assays measuring TSH binding may not be able to discriminate between malignant and benign dog thyroid tumours. TSH receptor number or affinity may be related to the functional variability of thyroid neoplasms.(ABSTRACT TRUNCATED AT 250 WORDS)

The canine p53 gene is subject to somatic mutations in thypoid carcinoma.

In many different types of tumors in man and mouse, p53 is the tumor suppressor gene most frequently affected by a combination of somatic mutation and loss of the wildtype allele. In order to develop a molecular tool to study the genetic evolution of tumors in the dog, we have cloned an evolutionary conserved part of the canine homologue of p53. The isolated genomic segment, 534 bp in length, contains the 3' half of exon 5, the complete exon 6 and the 5' half of exon 7, as well as the intronic intervening sequences. The gene organization of this segment shows strong homology to that published earlier for a number of other species, including man, mouse, and Xenopus laevis. This conservation is apparent at the DNA sequence level, as well as at the deduced aminoacid sequence level. mRNA expression can be detected at low levels in normal tissues with increased mitotic activity, and in the Madin-Darby canine kidney cell line. A-->G T transversion was found in 1 out of 23 investigated primary thyroid carcinomas at a position corresponding to codon 174 in the human p53, and was predicted to give rise to an aminoacid substitution in the protein. These results suggest that p53 plays a role in the development of malignancy in the dog, in a way comparable to that in man.

Flow-cytometric DNA ploidy analysis in primary and metastatic canine thyroid carcinomas.

DNA ploidy was measured by flow cytometry in 36 primary malignant thyroid neoplasms (including 6 bilateral tumours which were considered as separate neoplasms) from 30 dogs. In addition, DNA ploidy was determined in local recurrences in 3 dogs, and in 18 metastatic sites from 14 dogs. Aneuploidy was found in 21 of 36 (58%) primary sites. Eighteen of the 21 (86%) aneuploid tumours contained hypodiploid cell populations, with 12 having single hypodiploid peaks, and 6 being multiploid. Three other tumours had single aneuploid peaks with a DNA index (DI) greater than 1.0. The DIs in local recurrences were identical to those in the original neoplasms. Ploidy status (diploid vs. aneuploid) was identical in primary and metastatic sites in 10 out of the 14 dogs. Aneuploidy was more frequent in carcinomas from dogs with distant metastases (78%) than from dogs with less advanced stages of disease (53%), although this difference was not significant. There was no significant correlation between DNA ploidy and histopathological variables. From the strikingly high frequency of hypodiploidy in canine tumours, it is concluded that ploidy evolution in canine neoplasms may differ from that in human tumours.

Tubal factor pathology caused by Chlamydia trachomatis: the role of serology.

One of the causes of infertility in females is tubal pathology as a result of pelvic inflammatory disease (PID) caused by Chlamydia trachomatis (Ct). Diagnosis and identification of patients is hampered by the lack of rapid, easy, sensitive and specific methods. The introduction of Ct-specific enzyme-linked immunosorbent assay (ELISA) tests, based upon synthetic peptides may subsequently increase the sensitivity and specificity for the detection of tubal factor infertility caused by Ct. In order to determine the value of these tests for serological diagnosis of Ct infections, we evaluated several commercially available assays (C. trachomatis enzyme immunoassay (EIA), Labsystems (CtL); SeroCT, Savyon (CtS); pELISA, Medac (CtMp); and a reference assay rELISA, Medac (CtMr)) in two study populations. The first group consisted of 134 female patients with infertility problems. Tubal factor infertility was observed in 85 of these patients (63%). A higher % positivity was found for Ct-specific IgG for the CtL, CtS and CtMp, 41% vs 10%, 57% vs 18% and 55% vs 25% respectively as compared to patients with infertility due to other problems. A similar trend was observed for Ct-specific IgA. The specificity of Ct-specific IgA and IgG in this patient group varied between 92 to 98% and 76 to 90%, respectively. The second group consisted of 107 consecutive gynaecology patients with fertility problems or suspected PID. In this particular patient group, the specificity of the peptide based tests were around 80% and 90% for Ct-specific IgA and 75% and 85% for Ct-specific IgG, respectively. The negative predictive values exceeded 90%, while the positive predictive values varied from 30% to 47% for Ct-specific IgA and was around 30% for Ct-specific IgG. Testing Ct-specific IgG had no additional value above Ct-specific IgA alone. We conclude that the new synthetic peptide-based EIA tests are able to detect species-specific antibodies, which are correlated to (active) infection, and that in particular IgA may be useful in the serodiagnosis of tubal factor infertility caused by C. trachomatis, and will contribute in simplifying the work-up in patients with infertility.

Sensitivity and specificity of three new commercially available Chlamydia trachomatis tests.

In order to determine the value of new Chlamydia trachomatis (Ct) specific tests for routine serological diagnosis of Ct infections, we evaluated several commercially available assays (C. trachomatis enzyme immunoassay (EIA), Labsystems (CtL); SeroCT, Savyon (CtS); pELISA, Medac (CtMp)) in various study populations. The prevalence of C. trachomatis-specific IgA antibodies in a blood donor population (n = 443) as determined by the peptide based tests CtL, the CtS and the CtMp was 5%, while for IgG antibodies this was 6% (CtL and CtS) and 12% (CtMp) respectively. Prevalence was negatively correlated with age, concording with C. trachomatis specificity. None of the three tests showed significant titre rises in serum samples taken from patients with a proven infection of Chlamydia pneumoniae (n = 22), indicating species-specificity for all three tests. In patients with a polymerase chain reaction proven (n = 324) Ct infection, 75%, 70% and 68% were positive for IgG and 45%, 38% and 47%, positive for IgA as determined by the CtMp, CtL and CtS respectively. We conclude that the new synthetic peptide-based EIA tests are able to detect species-specific Ct antibodies, which are strongly correlated to (active) infection.

Ability of veterinary pathologists to diagnose classical swine fever from clinical signs and gross pathological findings.

Clinical signs recorded in a classical swine fever (CSF)-suspect situation and the results of the subsequent post-mortem examination (PME) from swine submitted to post-mortem during the 1997-1998 CSF epidemic in the Netherlands, were presented in an experiment as anonymous cases (without knowledge of the actual infection status of the submission) to five veterinary pathologists for their judgment: CSF-suspect or non-suspect. It was presented to them in two hypothetical situations: country was free of CSF for 5 years and CSF was detected in the country 2 weeks ago. Subsequently, their judgment was compared to the gold standard (infection status of the submission on the basis of an immunofluoresence assay on tissue samples) and the sensitivity (Se) and specificity (Sp) of clinical diagnosis was estimated. Furthermore, intra- and inter-pathologist agreement on pathological diagnosis was measured. Adding information on clinical signs to information on PME resulted in a significant increase in median Se and a significant decrease in median Sp for a clinical judgment by pathologists. Also, median Se was significantly higher-and Sp significantly lower-for a clinical judgment by pathologists in a situation in which CSF was detected 2 weeks ago in the country compared to the situation in which the country had been free of CSF for 5 years. Apparently, the internal threshold of pathologists is severely adjusted depending on the actual disease situation in a country. Intra-rater agreement ranged from fair to almost perfect for a clinical diagnosis on the basis of PME. There was considerable variation between pathologists, especially if clinical diagnosis was based on the combined information on PME and clinical signs observed in the herd. Inter-rater agreement was substantial for the diagnosis based on information on PME. Inter-rater agreement was considerably lower for the diagnosis based on the combined information on PME and clinical signs observed in the herd.

Lobular capillary haemangiomas in two calves.

Lobular capillary haemangiomas in the gingiva near the mandibular incisor region of two 6-month-old calves are described.

Dystrophy-like myopathy in the cat.

Two cases of feline myopathy are described which were associated with moderate locomotor disturbances. At necropsy, a pale coloured skeletal musculature was found with severe hypertrophy of diaphragmatic musculature. Histologically, the myopathy was characterized by varying fibre diameter, internal nuclei, moderate degeneration and necrosis of solitary muscle fibres and slight to moderate endomysial and perimysial fibrosis. Only very few regenerating muscle fibres were present. The histological findings are compatible with those found in human muscular dystrophies and so this feline myopathy may be considered as a dystrophy-like myopathy.

Lobular capillary haemangiomas in young horses.

Seven cases of cutaneous haemangiomas in young horses are described, characterized by scattered, well demarcated but unencapsulated lobules, consisting of capillary sized vascular structures, separated by strands of fibrous tissue. In one case, a more cellular variant was distinguished with minor vasoformative capacity. The morphological features of these haemangiomas are compatible with bovine cutaneous angiomatosis and human granuloma pyogenicum. The lesions are considered to be vascular tumours rather than hamartomas or granulation tissue and "lobular capillary haemangioma" is suggested to designate this type of haemangioma. In one tumour, cavernous vascular structures were also present in addition to the capillary structures and so this tumour is considered to be a combined capillary and cavernous haemangioma.

Canine capillary and combined capillary-cavernous haemangioma.

Six cases of capillary haemangiomas in the skin and one case in the tongue of dogs are described. There was no breed or age predisposition (average age was 8.9 years), but there was a prevalence of male dogs. In three cases, cavernous haemangioma-like structures were also found, so these tumours are considered to be combined capillary and cavernous haemangiomas.

Caprine peripheral neuroblastomas: structural and ultrastructural features, intermediate filaments profile and neuro-endocrine characterization.

In this report the morphological and immunohistochemical features with respect to intermediate filament proteins and the neuroendocrine nature of bilateral neuroblastomas possibly originating from the adrenal medulla in a goat kid are presented. Histologically, the tumours were composed of small, round, blue cells organized in highly cellular nests and sheets separated by fibrovascular septa. Isolated cells mimicking the morphological features of neurones were observed in both tumours. Ultrastructurally, dense-core neurosecretory granules, about 100 nm in diameter, and irregularly organized neurotubular networks were seen. The small tumour cells were only labelled by vimentin, while the neurone-like cells were labelled by both neurone-specific enolase and synaptophysin. The lack of staining of the tumour cells by the neurofilament proteins antiserum, which was also observed in a human neuroblastoma (used as positive control) has been previously reported and is probably the result of the tissue processing and/or the poor differentiation stage of the tumours. The vimentin labelling of tumour cells could be explained by the poor differentiation stage of the tumours, since vimentin is the only intermediate filament protein in presumptive neuroblasts, being replaced by neurofilament proteins in later stages of neurogenesis.

The use of keratin antisera in the characterization of a feline thymoma.

A cystic mass in the anterior mediastinum of a 9-year-old female European Shorthair cat was classified as a lymphocytic thymoma based on its histopathological features which were in accordance with those reported in the literature concerning feline thymomas. The application of a polyclonal keratin antiserum and monoclonal keratin antisera RCK 102, RKSE 60 and RGE 53 resulted in staining of foetal feline thymic cells, oesophageal epithelial cells as well as numerous stellate tumour cells and Hassall's corpuscles. As a result, the epithelial origin of the neoplastic cells could be established and the classification of thymoma confirmed. The results indicate the value of keratin antisera in the differentiation of thymoma and non-epithelial tumours in the anterior mediastinum.

Prevalence of exposure and infection of Lawsonia intracellularis among slaughter-age pigs.

The extent of clinical or subclinical infection associated with Lawsonia intracellularis within Dutch pig herds was uncertain. A case-control study of slaughter age pigs was used to study natural infection within Dutch herds and to compare diagnostic methods. From six case herds where clinical disease had been identified recently, and six disease-free herds, 40 pigs of slaughter-age were examined postmortem. The diagnostic methods used were: serology, gross examination, Haematoxylin and Eosin stain (HE), Warthin-Starry silver stain, Lawsonia-specific indirect immunoperoxidase of the ileum, and PCR of ileum mucosa and colon contents. There were 59% seropositive pigs in case herds and 26% seropositive pigs in control herds. Using immunohistochemistry, 57% of case herds and 46% of control herds were bacteria positive in the ileum mucosa. It was concluded that a majority of Dutch herds contain L. intracellularis infected finisher pigs. In some herds this is associated with clinical outbreaks of acute haemorrhagic enteropathy but in other herds no clinical disease is apparent. Many seropositive pigs in herds without clinical disease had evidence of Lawsonia antigen in sites other than the apical cytoplasm of proliferating epithelial cells, particularly the supranuclear region. It was uncertain whether to classify these pigs as having "recovered" from an infection or whether they have a sub-clinical or chronic form of the disease. We concluded that PCR examination of faeces and serology probably provide more specific results than gross examinations at slaughter, and that a monoclonal antibody-based examination of ileum mucosa should be the accepted screening method for this infection.