Role of mastitis in on-farm deaths of Finnish dairy cows.

According to our recent necropsy-based study, mastitis is the most common underlying diagnosis of on-farm deaths in Finnish dairy cows. However, it remained unanswered to what extent mastitis has contributed to death of all necropsied cows. In the present study, based on histopathology we detected one third of the necropsied dairy cows having active inflammatory udder lesions (n = 110). The role of mastitis varied and was interpreted by causes of death (underlying, intermediate, immediate, other significant). Mastitis was most commonly either the underlying (28%) or both immediate and underlying cause of death (48%), and only seldom the immediate (4%) or intermediate (4%) cause of death. Mastitis occurred either as the only cause leading to death (mastitis only, MO, 39%), or with many other contributing diseases (multiple diseases, MD, 61%) which were interacting with mastitis and together leading to death. Between these groups (MO vs. MD), time of mastitis occurrence during lactation, producer-reported duration of illness, clinical signs, and medication differed, as well the histopathological severity of mastitis. The cases, where mastitis was the only initial insult, occurred evenly throughout the entire lactation, but the cases with many interacting diseases clustered in early lactation. In multiple diseases -cases mastitis occurred concurrently with metritis (31%), aspiration pneumonia (24%), acute trauma/dystocia (15%), or with other diseases, such as ketosis, hepatic lipidosis, rumenitis, and abomasal diseases. For a pathologist, the gross mastitis diagnosis was most challenging at the beginning of the lactation, especially if inflammation was mild to moderate, suggesting the value of histopathological examination being highest at that time. Also, producers reported mastitis signs less frequently if cow had many simultaneously occurring diseases. Therefore, even if clinical signs of other diseases are present, the udder should be considered a potential cause of illness, and it should be examined, especially in dry and transition period cows.

Necropsy-based study on dairy cow mortality-Underlying causes of death.

The worldwide problem of increasing dairy cow mortality is widespread in modern production systems, it causes economic losses, and indicates problems with herd health and welfare. Most studies on causes of dairy cow mortality are limited as they are based on secondary register data, or questionnaires for producers or veterinarians, and neither necropsies nor histopathologic analyses are usually performed. For this reason, no definite causes for dairy cow deaths have been determined making it difficult or impossible to implement effective preventive measures. The objectives of this study were to (1) determine the causes of on-farm mortality of Finnish dairy cows, (2) determine the usefulness of routine histopathologic analysis in bovine necropsies, and (3) assess how reliable producers' perception about the cause of death is. Underlying diagnoses of on-farm deaths were determined through necropsy of 319 dairy cows at an incineration plant. The necropsy data were combined with background information obtained from online questionnaires covering cow and herd records. Mastitis was the most common underlying diagnosis of death (26.6%), followed by digestive disorders (15.4%), other known disorders (13.8%), calving-associated disorders (12.2%), and locomotion disorders (11.9%). The underlying diagnoses of death varied during different stages of lactation and with parity. A large proportion of the study cows (46.7%) died during the first 30 d after calving, and of those, 63.6% died during the first 5 d. A routine histopathologic analysis was performed in every necropsy, and it changed the preliminary gross diagnosis in 18.2% of the cases. Producers' perception about the cause of death agreed with the necropsy-based underlying diagnosis of death in 42.8% of the cases. It was most consistent for mastitis, calving disorders, locomotion diseases, and accidents. In cases where producers had no insights about the cause of death, necropsy revealed the final underlying diagnosis in 88.2% of cases, demonstrating the usefulness of necropsy. Based on our findings, necropsies provide useful and reliable information to develop control programs for cow mortality. Including routine histopathologic analysis in necropsies, more accurate information may be obtained. Furthermore, targeting preventive measures on transitional cows may be most effective, as the number of deaths was highest at this time.

Molecular identification of Taenia spp. in the Eurasian lynx (Lynx lynx) from Finland.

Cestodes of the genus Taenia are parasites of mammals, with mainly carnivores as definitive and herbivores as intermediate hosts. Various medium-sized cats, Lynx spp., are involved in the life cycles of several species of Taenia. The aim of the present study was to identify Taenia tapeworms in the Eurasian lynx (Lynx lynx) from Finland. In total, 135 tapeworms from 72 lynx were subjected to molecular identification based on sequences of 2 mtDNA regions, the cytochrome c oxidase subunit 1 and the NADH dehydrogenase subunit 1 genes. Available morphological characters of the rostellar hooks and strobila were compared. Two species of Taenia were found: T. laticollis (127 samples) and an unknown Taenia sp. (5 samples). The latter could not be identified to species based on mtDNA, and the rostellar hooks were short relative to those described among other Taenia spp. recorded in felids from the Holarctic region. In the phylogenetic analyses of mtDNA sequences, T. laticollis was placed as a sister species of T. macrocystis, and the unknown Taenia sp. was closely related to T. hydatigena and T. regis. Our analyses suggest that these distinct taeniid tapeworms represent a putative new species of Taenia. The only currently recognized definitive host is L. lynx and the intermediate host is unknown.

Botfly Cephenemyia ulrichii larvae in the nasal cavity and nasopharynx of a dog with respiratory signs.

A Finnish Lapphund dog with acute upper respiratory signs and gagging was presented at veterinary clinic. During rhinoscopy, ten 1- to 2-mm long, actively moving larvae were found in the dog's nasal cavity and nasopharynx and identified as Cephenemyia ulrichii (Diptera: Oestridae). This moose (Alces alces) parasite is widespread in Finland but has not been reported before from an accidental canine host. Clinical signs resolved with imidacloprid/moxidectin spot-on formulation.

Sexual Orientation Among Unmarried Final Year University Students in Osun State: Distribution, Determinants and Level of Satisfaction.

BACKGROUND: Sexual Orientation is an enduring pattern of romantic or sexual attraction, with the attractions broadly incorporated under heterosexuality and non-heterosexuality. Societal and other factors seem to affect individuals' perception of their self-identified orientation. Thus, this study sets out to document the distribution of sexual orientation, level of self-reported satisfaction, and factors associated with sexual orientation among unmarried university students. METHODS: A cross-sectional study involving 550 respondents sampled using a multi-stage sampling technique. Data collected via a self-administered questionnaire and analyzed via IBM SPSS version 20.0. RESULTS: Most respondents (93.6%) were heterosexuals, 4.7% bisexuals, 1.3% lesbian women and 0.40% gay men. More heterosexuals (80.4%) were fully satisfied with their self-identified sexual orientation compared to 57.2% of non-heterosexuals. Respondents who had religious upbringing and those whose parents were currently married were less likely to have sexual orientation outside of heterosexuality. CONCLUSION: The most prevalent sexual orientation was heterosexuality, with more heterosexuals also expressing satisfaction than others. With the level of dissatisfaction expressed by those who are not heterosexuals as well as the factors associated with heterosexuality, it is plausible to conclude that societal values and/or prejudices constitute major factors in individuals self-identifying with some particular sexual orientation and the satisfaction derived therein.

High mobility group box-1 (HMGB-1) in osteoarthritic cartilage.

OBJECTIVES: Nucleosomal high mobility group box-1 (HMGB-1) is translocated and released from necrotic and activated cells as an endogenous danger signal (alarmin) and cytokine. It was hypothesised that it plays a role in osteoarthritis (OA). characterised by cellular activation, inflammation and enchondral bone formation. METHODS: Bovine knee joint samples, collected from culled animals, were scored using histologic/histochemical grading to intact looking, mild, moderate or severe and immunohistochemically stained for HMGB-1. Chondrocyte pellets, produced from human bone marrow-derived mesenchymal stem cells and stimulated with tumour necrosis factor-a (TNF-alpha), were similarly stained. RESULTS: In healthy looking OA cartilage chondrocyte nuclei were usually HMGB-1 negative and in mild OA staining was restricted to nuclei. In moderate OA lesions HMGB-1 was also seen in the cytoplasm and occasionally pericellular matrix and in severe OA lesions often also in intra- and inter-territorial matrix. The tidemark in healthy cartilage did not contain HMGB-1, which however was seen at this interface as linear deposits even in intact-looking and mild OA lesions, as multiple wave-like deposits in moderate and as heavy granular deposits in severe lesions. TNF-alpha stimulation of chondrocytes caused translocation of HMGB-1 from the nucleus to the cytoplasm. CONCLUSIONS: In resting chondrocytes tight nucleosomal HMGB-1 binding might cause steric hindrance of immunostaining. TNF-alpha- or OA-mediated activation leads to nuclear staining and cytoplasmic translocation. Advancing OA leads to increasingly intense extra-/pericellular deposition of HMGB-1 alarmin, indicating local chondrocyte activation and/or necrosis. In particular, HMGB-1 at the tidemark might play a role in the pathological thickening of subchondral bone plate/osteophyte formation.

Central primitive neuroectodermal tumour with ependymal differentiation in a dog.

A central primitive neuroectodermal tumour (PNET) with ependymal differentiation was identified in an 18-month-old female Australian terrier dog that died following signs of central nervous system disease. A large space-occupying haemorrhagic mass replaced the thalamus and part of the hypothalamus of the brain. Microscopical examination revealed a tumour formed by a compact sheet of small cells that contained ependymal and perivascular pseudorosettes. Immunohistochemically, the neoplastic cells variably expressed glial fibrillary acidic protein, neuron-specific enolase, vimentin and cytokeratin, consistent with divergent differentiation of the tumour. This case is thought to represent the first description of PNET with ependymal differentiation in a dog.

Expression of vascular endothelial growth factor receptors coincide with blood vessel in-growth and reactive bone remodelling in experimental intervertebral disc degeneration.

OBJECTIVE: To analyze immunohistochemically the localization of the VEGF receptors in experimental intervertebral disc degeneration tissues in a pig model. MATERIALS AND METHODS: In six domestic pigs, the cranial bony endplate of the L4 vertebra were perforated into the nucleus pulposus. Three months postoperatively, the animals were sacrificed and the experimental and control vertebrae, complete with intervertebral discs, were excised and subjected for immunohistochemical staining of vascular endothelial growth factor receptors (VEGFR) along with VEGF - A, -C, -D and blood and lymphatic vessel markers vWF and LYVE-1. RESULTS: The results of immunohistochemical analysis of experimental samples showed VEGFR-1 (Flt-1) expression in intervertebral disc and all paradiscal tissues studied. In control samples expression of VEGFR-1 was lower and absent in the intervertebral discs. Comparatively less of VEGFR-2 (KDR/Flk-1) and VEGFR-3 (Flt-4) than VEGFR-1was found in degenerated intervertebral discs and paradiscal tissues. In contiguous control intervertebral discs and control paradiscal tissues VEGFR-2 and-3 receptors were expressed to a lower extent than in experimental tissues or were even totally absent. Also growth factors VEGF-A, -C, -D, as well as von Willebrand factor and to a much lower extent LYVE-1 were differently expressed in experimental and control intervertebral discs. CONCLUSION: In experimental intervertebral disc degeneration, VEGF receptors were expressed in the damaged disc and paradiscal tissues. In the same tissues, VEGF-A, -C, and -D, signs of blood and lymphatic vessel in-growth and reactive/adaptive vertebral bone remodelling were found.

Infertile boars with knobbed and immotile short-tail sperm defects in the Finnish Yorkshire breed.

In the period 1996-2006 two specific sperm defects, the knobbed acrosome (KA) defect and the immotile short-tail sperm (ISTS) defect, showed a strong negative association with fertility in Finnish breeding boars. In this study, we examined the incidence of these two sperm defects in two pig breeds, their effects on fertility and their associations with sperm morphology and testicular histology. Semen samples from 2048 (1097 Yorkshire, 951 Landrace) boars were collected. None of the Landrace boars revealed either the KA defect or the ISTS defect. Of the Yorkshire boars, 0.8% were afflicted with the KA defect and 7.6% with the ISTS defect. Boars diagnosed with the ISTS defect produced no litters. Fertility data were available from two artificially inseminated (AI) boars and six farm breeding boars affected with the KA defect. Breeding boars with 45-81% knobbed spermatozoa (n = 6) did not produce any litters out of 71 sows bred. AI boars with 25-30% knobbed spermatozoa had a poor non-return rate (on average 47% compared with 85% for normal control boars) and produced small litters, on average 2.5 piglets less than other boars of the same breed. Morphometry of testicular tissue and distribution of different cells in the seminiferous tubules were examined in nine boars. Boars with the KA defect had a smaller diameter of the seminiferous tubules (p < 0.05) and a lower number of Sertoli cells (p < 0.05) than controls. ISTS boars, in turn, had a significantly lower number of elongated spermatids (p < 0.05), and they also produced on average only 12% of the spermatozoa of normal boars. The ISTS defect is a manifestation of an autosomal recessive disease caused by an insertion in the KPL2 gene in porcine chromosome 16. Although we tried to map the KA defect, its aetiology remains unclear.

Non-tuberculous Mycobacteria can Cause Disseminated Mycobacteriosis in Cats.

Mycobacteriosis caused by non-tuberculous mycobacteria (NTM) is a rising concern in human medicine both in immunocompromised and immunocompetent patients. In cats, mycobacteriosis caused by NTM is considered mostly to be a focal or dermal infection, with disseminated disease mostly caused by Mycobacterium avium. We describe three cases of disseminated mycobacteriosis in cats, caused by Mycobacterium malmoense, Mycobacterium branderi/shimoidei and M. avium, with no identified underlying immunosuppression. In all cases, extracellular mycobacteria were seen in the pulmonary epithelium, intestinal lumen and glomerular tufts, which could affect the shedding of the organism. The present study highlights the importance of mycobacteriosis as a differential even in immunocompetent animals. Considering the close relationship of owners and pets and the potential presence of free mycobacteria in secretions, cats should be considered as a possible environmental reservoir for mycobacteria.

Providing high-quality research training for veterinary pathologists in Europe.

Despite their key role in a wide range of fields relating to animal and public health, there is currently a lack of veterinary pathologists in Europe. In 1999, to help address the problem, the European College of Veterinary Pathologists (ECVP) and the European Society of Veterinary Pathology (ESVP) established a joint Education Committee. In this Special Article, Professor Anja Kipar and colleagues, all members of the committee, describe the ECVP/ESVP Summer Schools in Veterinary Pathology programme, which aims to provide high-quality research training for veterinary pathologists from all over Europe and beyond.

Temporal changes in concentrations of serum amyloid-A and haptoglobin and their associations with weight gain in neonatal reindeer calves.

Age-related changes in serum concentrations of two acute phase proteins (APPs), haptoglobin (Hp) and serum amyloid-A (SAA) were investigated in newborn reindeer calves. Repeated blood samples were obtained from 51 reindeer calves at ages 0-32 days (2-4 samples from each calf). An increase of SAA concentrations was observed during the first 2 weeks of life. However, by the end of the observation period, SAA concentrations had decreased to levels below those of the first week. Serum Hp concentrations increased throughout the observation period. SAA concentrations in the second week had a negative association with weight gain during the entire study period (4 months). These time-related changes in APP concentrations suggest that these proteins have a role in the defence and adaptation mechanisms of newborn reindeer calves. Possible reasons for these changes include the presence of APP mediators in the colostrum, exposure to environmental pathogens after birth and age-related changes in hepatic synthesis of APP.

Canine intestinal helminths in Finland: prevalence, risk factors and endoparasite control practices.

In this survey, the prevalence of canine gastrointestinal helminths in Finland was investigated by coprological examination (n = 541) and possible risk factors for helminth infections in dogs were analysed. In addition, the dog owners (n = 296) completed a questionnaire about use of anthelmintics, sources of information about parasites and antiparasitic treatments and reasons for choosing the drugs. The prevalence of gastrointestinal helminths was 5.9%. Eggs from four different species were identified in the faecal samples. Toxocara canis eggs were present in 17 dogs (3.1%), Uncinaria stenocephala eggs in 14 dogs (2.6%) and Diphyllobothrium latum eggs in 2 dogs (0.4%). Moreover, one sample contained eggs of Trichuris vulpis (0.2%). Kennel housing and visits abroad were identified as risk factors for T. canis and U. stenocephala infections. Most dogs (86.0%) received anthelmintic treatment at least once a year. Hunting dogs were dewormed least; one-third was treated less than once a year. Approximately, half of the owners occasionally changed the anthelmintic used. The most important trait of the anthelmintic was its broad spectrum, fenbendazole being the most commonly chosen. Veterinarians, dog magazines and dog breeders were the predominant sources of information concerning parasites and deworming strategies.

Testing of amplified fragment length polymorphism (AFLP) technique as a tool for molecular epidemiology of Trichinella nativa.

A total of nine Trichinella nativa isolates were compared by amplified fragment length polymorphism (AFLP). Four hundred nanograms of genomic DNA from a pool of 10--20 larvae were digested using HindIII and MseI restriction endonucleases. Of the 16 primer combinations initially tested, Hind-C and Mse-C primers showed rich polymorphism with approximately 40--90 bands in the range of 30--270 bp. Genetic similarities were estimated visually. AFLP provided discriminatory banding patterns and may therefore be used as a method for detecting variation in T. nativa populations. However, the heterogeneous patterns obtained from pooled samples emphasize the need for further development of the sampling and numerical analysis of the patterns for epidemiological and taxonomical interpretation.

Spatial variation of Trichinella prevalence in rats in Finnish waste disposal sites.

Trichinellosis is 1 of the most widespread parasitic zoonoses in the world and can be lethal to humans. Trichinella spp. are also parasites of considerable economic importance. Because rats may play a role in the transmission of trichinellosis to swine and farmed wild boar, 767 brown rats (Rattus norvegicus Berkenhout) from 13 Finnish waste disposal sites were examined for Trichinella spp. by a HCl-pepsin digestion method. Trichinella spp. were found to be a common parasite in trapped rats (overall prevalence, 19%) detected in 12 of 13 dumps. Significant differences were observed between sites in the prevalence (0-49%) of Trichinella spp. Female rats were more often and more heavily infected than males, but age was not shown to be a risk factor for trichinellosis. In addition, positive correlation was demonstrated between rat population density and prevalence. Trichinella spiralis was identified by multiplex polymerase chain reaction in 28 rats. The median density of infection was 42 (range, 0.5-6,925) larvae/ g of host tissue, but neither the occurrence nor the density of the parasite was related to the physical condition of the animal.

Neither hippurate-negative Brachyspira pilosicoli nor Brachyspira pilosicoli type strain caused diarrhoea in early-weaned pigs by experimental infection.

A hippurate-negative biovariant of Brachyspira pilosicoli (B. pilosicolihipp-) is occasionally isolated in diarrhoeic pigs in Finland, often concomitantly with hippurate-positive B. pilosicoli or Lawsonia intracellularis. We studied pathogenicity of B. pilosicolihipp- with special attention paid to avoiding co-infection with other enteric pathogens. Pigs were weaned and moved to barrier facilities at the age of 11 days. At 46 days, 8 pigs were inoculated with B. pilosicolihipp- strain Br1622, 8 pigs were inoculated with B. pilosicoli type strain P43/6/78 and 7 pigs were sham-inoculated. No signs of spirochaetal diarrhoea were detected; only one pig, inoculated with P43/6/78, had soft faeces from day 9 to 10 post inoculation. The pigs were necropsied between days 7 and 23 after inoculation. Live pigs were culture-negative for Brachyspira spp., but B. pilosicolihipp- was reisolated from necropsy samples of two pigs. The lesions on large colons were minor and did not significantly differ between the three trial groups. In silver-stained sections, invasive spirochaetes were detected in colonic mucosae of several pigs in all groups. Fluorescent in situ hybridisation for genus Brachyspira, B. pilosicoli and strain Br1622 was negative. However, in situ detection for members of the genus Leptospira was positive for spirochaete-like bacteria in the colonic epithelium of several pigs in both infected groups as well as in the control group. L. intracellularis, Salmonella spp., Yersinia spp. and intestinal parasites were not detected. The failure of B. pilosicoli strains to cause diarrhoea is discussed with respect to infectivity of the challenge strains, absence of certain intestinal pathogens and feed and management factors.

Trophozoite-to-cyst ratio increases during recovery from Pneumocystis carinii pneumonia in rats.

Pneumocystis carinii has two developmental stages: the trophozoite and the cyst. The differential specificity of monoclonal antibodies SMoAb (cysts only) and CMoAb (cysts and trophozoites) was used to analyse the trophozoite-to-cyst (T: C) ratio during recovery from P. carinii pneumonia (PCP) in a dexamethasone model in Wistar rats. In tissue homogenates the cysts decreased from week 0 to week 4 (4.10 +/- 1.57 10(6)/g vs 0.069 +/- 0.021 10(6)/g, p < 0.001), whereas the trophozoites decreased much less, leading to a change in T: C ratio from week 0 and 1 (72 +/- 33 and 49 +/- 14, respectively) to week 2 and 4 (1752 +/- 652 and 1163 +/- 482, p < 0.001). A similar tendency was found in immunofluorescence-labelled histological samples quantitated with the VIDAS image analyser. These results demonstrate a hitherto unrecognized and significant change in T: C ratio during recovery from PCP. This change could not be monitored using bronchoalveolar fluid, probably due to adherence of the trophozoites to the alveolar wall. Changes in T: C ratio indicate that in particular the cyst form of P. carinii is sensitive to the host response mounted during recovery, and support the theory that trophozoites probably also multiply in an extracystic asexual fission.

An outbreak of trichinellosis in farmed wild boar in Finland.

Nine farmed wild boar out of 25 slaughtered from a single farm were condemned at meat inspection because of trichinellosis. With RAPD-PCR, Trichinella spiralis was identified in all positive wild boar. Out of the available serum samples (n=7), all wild boar which had failed the meat inspection showed seroconversion in ELISA and Western blotting, as did one additional animal which had passed the inspection. The animals became infected during an invasion of rats from an improperly closed dump near the farm. Unfortunately, by the time trichinellosis was discovered in the wild boar, the invasion had already been brought under control; thus, no samples from rats were available. However, having lived through the rat invasion was shown to be a risk factor for trichinellosis in wild boar (relative risk, RR=6.3). In wildlife samples from surrounding areas, sylvatic trichinellosis was found to be very common (74%; n= 19 red foxes). Intriguingly, the prevalent species in trichinella-positive foxes differed from that in wild boar, Trichinella nativa and T. spiralis being found in 12 foxes and in one fox, respectively.

Ultrastructural localization of monoclonal antibodies against Pneumocystis carinii. Differentiation between developmental stage and host origin.

The ultrastructural immunolocalization of target antigens recognized by two monoclonal antibodies against Pneumocystis carinii was investigated using both human- and rat-derived organism. Labelling was seen using both cryo electron microscopy and postembedding in Epon after fixation in glutaraldehyde. The antibodies showed different host species- and developmental stage specificity. One of the antibodies reacted with the outer membrane of all developmental stages of human-derived but not rat-derived organisms, whereas the other reacted only with the cyst form. The distribution of the latter was similar to that reported for methenamine silver, a widely used cytochemical stain for the parasite, and reacted with both human- and rat-derived organisms. The antibodies described appear to be useful markers in studies on the differentiation of Pneumocystis carinii.

Dual effects of caspase-1, interleukin-1 beta, tumour necrosis factor-alpha and nerve growth factor receptor in inflammatory myopathies.

OBJECTIVE: To analyse the expression of factors potentially involved in skeletal muscle degeneration and regeneration in dermatomyositis (DM), systemic sclerosis (SSc), polymyositis (PM), systemic lupus erythematosus (SLE) and non-inflammatory myopathies. METHODS: Immunohistochemical staining of skeletal muscle biopsies (10 DM, 10 SSc, 10 PM, 10 SLE, 10 non-inflammatory myopathies) for tumour necrosis factor-alpha (TNF-alpha), interleukin-1 beta (IL-1 beta), activated caspase-1, pan-macrophage marker CD68, inducible nitric oxide synthase (NOS2) and nerve growth factor receptor (NGFR). TechMate staining robot and biotin-streptavidin protocol were used. RESULTS: Expression of TNF-alpha, IL-1 beta, caspase-1 and NOS2 was found in the cytoplasm and sarcolemma of dystrophic skeletal muscle fibres. TNF-alpha and IL-1 beta immunoreactive profiles were faint and few and close to satellite nuclei-containing regenerating muscle fibres both in inflammatory and non-inflammatory myopathies. NGFR expression was found in comparable areas. In non-inflammatory inherited myopathies more nuclei were caspase-1 immunoreactive whereas caspase-1 expression was rarely seen in inflammatory myopathies, implying regeneration of the affected muscle fibres. CONCLUSION: Prominent expression of the proinflammatory factors TNF-alpha, IL-1 beta and NOS2 and caspase-1 is associated with muscle fibre damage, albeit when expressed to a low degree these factors may, like NGFR, contribute to muscle regeneration and healing.