Effect of bovine seminal plasma on bovine endometrial epithelial cells in culture

The purpose of this study was to investigate the effect of seminal plasma (SP) from bulls of known fertility on bovine endometrial epithelial cells (bEEC) in culture. The bEEC from passage 5, approximately 5.0–13 × 10⁵ cells per flask, were challenged with SP from bulls of high or low fertility (n = 3 and 2, respectively) or PBS (control), at 1% (75 μl) or 4% (300 μl) and were incubated for 72 hr (n = 13 per challenge). Total cell number and viability of bEEC after challenge with 1% SP from either high‐ or low‐fertility bulls (75H or 75L, respectively) did not differ from controls. In contrast, challenge with 4% of SP from high‐ or low‐fertility bulls (300H or 300L) negatively affected bEEC cell number and viability. Challenge with 300 L had a greater adverse effect than 300H. These results suggest that the negative effect of bovine SP on bEEC is both dose‐dependent and fertility‐dependent.     

The polymorphisms of bovine melanocortin-3 receptor pseudogene

Mammalian melanocortin-3 receptor (MC3R) plays an important role in the central control of energy homeostasis, and several functional polymorphisms of mc3r have been detected. Interestingly, the bovine mc3r was a pseudogene, and its polymorphisms and function remain to be investigated. Single-strand conformation polymorphism (SSCP) showed 5, 2 and 3 genotypes in fragment F1, F2 and F3 of mc3r in seven cattle breeds, respectively. All genotypes revealed novel sequences. Three SNPs 657G > T, 756C > T, 822T > C were detected in fragment F1, five SNPs 1091T > C, 1133T > C, 1144C > T, 1259T > C and 1319G > A were detected in fragment F2, and two SNPs 1687G > A, 1860C > T were detected in F3. The SNPs in fragment F1 and F2 were located at exon 2. The five SNPs in fragment F2 demonstrated a tight linkage disequilibrium status. Variation detected here might have an impact on the function of bovine mc3r pseudogene.     

Enzootic bovine leukosis virus in Brazil

Summary A sero-epidemiological survey for antibodies to the glycoprotein of enzootic bovine leukosis virus showed that the infection is widely disseminated in the State of Rio de Janeiro, Brazil. Sera from 1,290 females and 154 males from 12 dairy herds were tested by the agar gel precipitin test. Seven hundred and one females (54.3%) and 68 males (44.2%) were found to have specific antibodies. These antibodies were demonstrated in all 7 age groups tested. The older age groups contained the highest percentage of reactors. The results are briefly discussed in relation to management practices and environmental conditions.

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El Virus De La Leucosis Bovina Enzootica En Brasil

Resumen Una encuesta suero-epidemiológica por anticuerpos contra la glicoproteina del virus de la leucosis bovina enzootica mostró que la infección está ampliamente diseminada en el Estado de Rio de Janeiro, Brasil. Sueros de 1290 hembras y 154 machos pertenecientes a 12 rebaños lecheros fueron examinados en la prueba de precipitación en agar. Se encontró que 701 hembras (54.3 por ciento) y 68 machos (44.2 por ciento) poseían anticuerpos específicos. Estos anticuerpos fueron demonstrados en todos los siete grupos de edad examinados. Los grupos de mas edad contenian un porcentaje mayor de reactores. Los resultados son discutidos brevemente en relación a las prácticas de manejo y a las condiciones ambientales locales.

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Virus De La Leucose Bovine Enzootique Au Bresil

Résumé Une enquête séro-épidémiologique pour la détection de la présence de la glycoproteine du virus de la leucose enzootique bovine a montré que cette affection est largement disséminée dans l'Etat de Rio de Janeiro, au Brésil. Des échantillons de sérum provenant de 1290 femelles et 154 mâles appartenant à 12 troupeaux laitiers ont été étudiés par précipitation en gélose—701 femelles (54,3 p. 100) et 68 mâles (44,2 p. 100) ont été reconnus comme possédant des anticorps spécifiques. Ces anticorps ont été mis en évidence dans la totalité des 7 groupes d'âges étudiés. Les groupes d'âges plus élevés contenaient le plus haut pourcentage d'animaux positifs. Les résultats sont brièvement discutés, dans leur relation avec les pratiques de l'élevage et les conditions de l'environnement.

     

Propagation of infectious bovine rhinotracheitis (bovine herpes-1) virus in murine primary cell cultures

Virus synthesis in BALB/C mouse lung and kidney primary cultures infected with infectious bovine rhinotracheitis (IBR) virus started between 6 and 8 hours after virus inoculation and reached a maximum titer of 5.5 log₁₀ plaque forming units (PFU) at 48 hours postinfection (PI). Cytopathic effect (CPE) in cell cultures occurred at 8–10 hours and over 90% of the cells had CPE by 48 to 72 hours PI. The bulk of the newly replicated virus (60–80%) was cell-associated as determined by plaque assay of extracellular and intracellular virus. Pulse-chase experiments demonstrated incorporation of radioactive precursors into viral DNA and protein macromolecules. Viral DNA synthesis was initiated between 2 and 4 hours PI and was maximum at 4 to 6 hours. Viral proteins were detected at 4 hours and peaked between 6 and 8 hours PI. Enzyme-linked immunosorbant assay (ELISA) confirmed synthesis of specific viral proteins, which gradually increased during the virus growth cycle. Abstract in French is given at the end of the article.

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Resume La synthèse du virus de la rhinotrachéite infectieuse bovine (virus IBR) sur des cultures primaires de poumon et de reins de souris (souche Balb/C) commençait entre 6 et 8 heures après inoculation du virus et atteignait un titre maximal de 5.5 log PFU (Plaque Forming Unit) 48 heures après infection. L'effet cytopathogenic (CPE) sur ces cultures cellulaires apparaissaient en 8 à 10 heures et plus de 90% des cellules montraient un CPE en 48 à 72 heures après infection. La détermination par la méthode des plages sous agarose, du virus extracellulaire et du virus intracellulaire, indiquait que 60–80% du virus synthétisé était associé avec les cellules. Les expériences de pulsechase demontraient l'incorporation des précurseurs radioactifs dans les protèines et ADN viraux. La synthèse virale d'ADN était initiée entre 2 et 4 heures après infection et était maximale en 4 à 6 heures. Les protéines virales étaient détectées en 4 heures avec un maximum de synthèse entre 6 et 8 heures après infection. Enzyme-linked immunosorbant assay (ELISA) confirmait la synthèse de protéines virales spécifiques qui montrait une augmentation graduelle pendant le cycle de replication du virus.

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Supported in part by grants 0831 from the Cooperative State Research Service of the US Department of Agriculture and published as contribution 85-160-J, Kansas Agricultural Experiment Station.     

Prenatal diagnosis of bovine citrullinaemia

Holstein/Friesian embryos, progeny of citrullinaemia heterozygotes, were transferred to recipient cows. The citrullinaemia genotype of the fetus/calf was established, at birth, by estimation of citrulline concentration in plasma or by analyses of argininosuccinate synthetase exon 5 sequences in dna from blood leucocytes. Between 115 and 125 days of gestation amniotic fluid was collected transabdominally from nine recipient cows. The mean citrulline concentration in amniotic fluid from fetuses unaffected with citrullinaemia was 28 μM, SD 9 and range 15 to 38 μM. Citrulline concentration was 107 and 130 μM in amniotic fluid from two fetuses homozygous for the citrullinaemia mutation. Only the normal bovine citrullinaemia sequence was detected in cells cultured from amniotic fluid taken from two homozygous normal and two heterozygous fetuses. Both normal and mutant sequences were found in cells cultured from amniotic fluid collected from three of five heterozygous fetuses. Only the mutant sequence was found in cells cultured from the two citrullinaemia affected fetuses. mhc class II DRB3 haplotyping revealed that in four instances cultured cells were from the surrogate mother, not the fetus, these included samples from two heterozygotes in which only the normal sequence was detected in amniotic fluid.     

Effect of caspase inhibitor Z-VAD-FMK on bovine sperm cryotolerance

The aim of this study was to evaluate the treatment of bovine semen with the pan-caspase inhibitor benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone (Z-VAD-FMK), before or after freezing on semen quality. After the initial assessment, sperm from 4 bulls were pooled (Experiment 1) and cryopreserved in BioXcell containing 0, 20 and 100 μM Z-VAD-FMK. After thawing semen viability, motility, membrane integrity, as well as DNA fragmentation and ΔΨm were evaluated. In Experiment 2, bovine frozen/thawed sperm were incubated for 1 hr with 0, 20 and 100 µM Z-VAD-FMK before assessing the semen quality. The treatment with Z -VAD-FMK before cryopreservation improved post-thawing sperm motility compared to the control group (p < .05), while no differences were recorded in sperm viability and membrane integrity among groups (on average 86.8 ± 1.5 and 69.1 ± 1.4, respectively). Interestingly, at the highest concentration, DNA fragmentation decreased (p < .05), while the percentage of spermatozoa with high ΔΨm increased (p < .05). The results of Experiment 2 showed that 1-hr treatment with Z-VAD-FMK did not affect sperm motility and viability (on average 63.4 ± 5.8 and 83.7.1 ± 1.2, respectively). However, Z-VAD-FMK improved sperm membrane integrity (p < .05) and at the highest concentration tested decreased the proportion of sperm showing DNA fragmentation (p < .05). No differences were recorded in the percentage of spermatozoa with high ΔΨm (on average 57.0 ± 11.4). In conclusion, the treatment with 100 µM of the caspase inhibitor Z-VAD-FMK before freezing increased bovine sperm mass motility and ΔΨm, while decreasing sperm DNA fragmentation. Treatment of semen after thawing with 100 µM Z-VAD-FMK improved sperm membrane integrity and reduced DNA fragmentation.     

Viral antibodies in bovine fetuses in Argentina

In order to establish the prevalence of viral infections of the bovine fetus in Argentina, a serological survey for antibodies against viral agents currently affecting cattle in this country was conducted. Antibodies against foot-and-mouth disease virus (FMDV), bovine herpesvirus-1 (BHV-1), bovine leukaemia virus (BLV), bovine rotavirus (BRV), bovine coronavirus (BCV), bovine viral diarrhoea virus (BVDV) and parainfluenza-3 (PI-3) were investigated in a total of 315 fetal serum samples. Conventional techniques were used: indirect immunofluorescence (FMDV, BHV-1, BVDv and BCV), radial immunodiffusion (BLV), ELISA (BRV) and haemagglutination inhibition (PI-3). Antibodies against BHV-1, BVDV and PI-3 were detected in samples from fetuses in the second and third trimester of gestation, with a prevalence of 1·21 per cent (two of 165), 2·03 per cent (four of 197) and 5·08 per cent (nine of 177), respectively. Either antibodies or non-antibody factors able to bind to BRV and Bcv antigens were detected with a prevalence of 2·44 per cent (five of 205) and 4·54 per cent (five of 110), respectively. In addition, 14·68 per cent of non-specific inhibitors of PI-3 mediated haemagglutination were found. No seropositives against FMDV and BLV were detected.     

Biomechanics of the fresh and conserved bovine pericardium

The use of biological membranes in surgeries is a reality for years, and one of the most used is the bovine pericardium, so the purpose of this research is to describe the bovine pericardium's biomechanics by comparing two directions of a test, one parallel to the longitudinal heart axis and one perpendicular. 20 adult bovine pericardium were tested for the maximum rupture force and rupture elongation, collecting four samples of each pericardium direction. In phase 2, eight pericardia were conserved for 4 months in a 98% glycerine solution, and the solution in which they were submerged was microbiologically analysed monthly. The Mann–Whitney test was used; there was a very significant difference between the perpendicular and parallel groups (p = .0001). The T test showed no significant difference for the rupture elongation (p = .0938). In pericardium preserved in glycerine, the outliers were removed regarding the maximum rupture force, and a Boxcox transformation was performed (λ = 0.25). Outliers were removed for the rupture elongation, and Bartlett's test (p = .7836), and Cramer–Von Mises (p = .5033) were performed and then, the analysis of variance (p < .0001), followed by the Tukey test at 5%. In the microbiological analysis, there was no presence of microorganisms during conservation. The research has shown that the pericardium collection direction influences its resistance and it can be stored in glycerine for 4 months without losing biomechanical characteristics.     

Phylogenetic analysis and characterization of Korean bovine viral diarrhea viruses

Thirty-six bovine viral disease viruses (BVDVs) were identified in bovine feces (n = 16), brains (n = 2), and aborted fetuses (n = 18) in Korea. To reveal the genetic diversity and characteristics of these Korean strains, the sequences of their 5′-untranslated regions (5′-UTRs) were determined and then compared with published reference sequences. Neighbor-joining phylogenetic analysis revealed that most of the Korean viruses were of the BVDV subtypes 1a (n = 17) or 2a (n = 17). The remaining strains were of subtypes 1b (n = 1) and 1n (n = 1). This analysis indicates that the 1a and 2a BVDV subtypes are predominant and widespread in Korea. In addition, the prevalence of BVDV-2 was markedly higher in aborted fetuses than in other samples and was more often associated with reproductive problems and significant mortality in cattle.     

Age-dependent patterns of bovine tuberculosis in cattle

Bovine tuberculosis (BTB) is an important livestock disease, seriously impacting cattle industries in both industrialised and pre-industrialised countries. Like TB in other mammals, infection is life long and, if undiagnosed, may progress to disease years after exposure. The risk of disease in humans is highly age-dependent, however in cattle, age-dependent risks have yet to be quantified, largely due to insufficient data and limited diagnostics. Here, we estimate age-specific reactor rates in Great Britain by combining herd-level testing data with spatial movement data from the Cattle Tracing System (CTS). Using a catalytic model, we find strong age dependencies in infection risk and that the probability of detecting infection increases with age. Between 2004 and 2009, infection incidence in cattle fluctuated around 1%. Age-specific incidence increased monotonically until 24–36 months, with cattle aged between 12 and 36 months experiencing the highest rates of infection. Beef and dairy cattle under 24 months experienced similar infection risks, however major differences occurred in older ages. The average reproductive number in cattle was greater than 1 for the years 2004–2009. These methods reveal a consistent pattern of BTB rates with age, across different population structures and testing patterns. The results provide practical insights into BTB epidemiology and control, suggesting that targeting a mass control programme at cattle between 12 and 36 months could be beneficial.     

Trace micro-nutrients may affect susceptibility to bovine tuberculosis in cattle

Bovine tuberculosis (bTB) is a continuing problem in British herds. Micro-nutrients are important for the maintenance of well-functioning immune system. The aim of this study was to determine whether the selenium, copper and vitamin B12 status of cattle was associated with Mycobacterium bovis (M. bovis) infection. Between 2002 and 2005, 200 cattle (43% dairy, mean age 4.6 years), reactors according to the standard interpretation of the tuberculin test, and 200 in-contacts (41% dairy, mean age 4.4 years) non-reactors, which had been in contact with cattle with bTB, were selected from herds in England and Wales. Levels of the seleno enzyme glutathione peroxidase (GSHPx), copper and vitamin B12 were measured in blood. Confirmation of bTB infection was made by bacteriological culture and histopathology following a detailed postmortem. Levels of selenium and copper were also measured in a random sample of 63 livers. bTB was confirmed by culture/histology in 23/200 (11.5%) of in-contacts and 110/200 (55%) of reactors. In blood drawn at recruitment, GSHPx was lower in cattle with confirmed bTB compared to other cattle (geometric means 59.7 u/mL versus 78.9 u/mL red blood cells (RBC), p < 0.01). Vitamin B12 was similar (geometric means 161.5 pmol/L versus 165.5 pmol/L, p = 0.62) and copper was similar (geometric means 14.4 μmol/L versus 14.1 μmol/L, p = 0.55). In logistic regression models including all micro-nutrients simultaneously and controlling for age, sex, animal production class, herd size, number of reactors, postmortem laboratory and seasonal trends, lower levels of GSHPx (adjusted OR 0.42, 95% CI 0.21–0.81 per 100 u/mL RBC, p = 0.01) and higher levels of copper (adjusted OR 1.69, 95% CI 1.21–2.36 per 5 μmol/L, p < 0.01) were associated with an increased risk of confirmed bTB but there was no association with vitamin B12. There was evidence for a stronger association between confirmed bTB and GSHPx in in-contacts (adjusted OR 0.21, 95% CI 0.06–0.79 per 100 u/mL RBC) compared to reactors (adjusted OR 0.50, 95% CI 0.21–1.23 per 100 u/mL RBC) (p = 0.08 for interaction). Lower liver copper was associated with a higher risk of confirmed bTB (adjusted OR 0.15, 95% CI 0.02–1.0 per 5000 μmol/kg dry mass, p = 0.05) but there was no association between liver selenium and bTB. Trace micro-nutrient status may affect susceptibility to M. bovis infection in cattle. Further studies are needed.     

Amyloidosis in the bovine kidney

After a survey of the most important literature on amyloidosis the results of the authors own studies on amyloidosis in the bovine kidney are reported. The macroscopical and microscopical features of the various types of bovine renal amyloidosis are described. In most cases predominantly medullary depositions occurred (types A, B, C and D). In a series of over 1000 cases only six kidneys with glomerular amyloid were found (type E). All these, however, contained some medullary amyloid as well. It is questionable whether the different types of kidney lesions contain different forms of amyloid or represent variants of the same disorder. Clinico-pathological studies may solve this problem.Medullary amyloid may be a subclinical disease. In clinically normal slaughter cattle it occurred in 2.7% of 1326 animals. In clinically normal dairy cattle (including young animals) it occurred in 2.6% of 378 animals and the incidence appeared to increase with age. In the medullary amyloid deposits different amyloid-containing cell types were found. The intracellular amyloid was arranged in parallel fibrils and was localized in lysosomes. Induced phagocytosis of bovine amyloid by rat macrophages appeared to result in similar vacuoles, indicating that phagocytosis may occur in the bovine cells. In the literature parallel intralysosomal amyloid has been interpreted as formation of amyloid in these vacuoles.An attempt is made to elucidate the pathogenesis of amyloidosis.

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Kurzfassung Nach der Beschreibung der wichtigsten Literaturangaben von Amyloidose werden die eigene Befunde von Amyloidose der Niere des Rindes besprochen. Die makroskopischen und mikroskopischen Eigenschaften der Nierenamyloidose des Rindes werden beschrieben. In den meisten Fällen gab es vor allem medulläre Ablagerungen (Typ A, B, C, und D). Bei über 1000 untersuchten Rindernieren wurden nur 6 Nieren mit hauptsächlich glomerulärem Amyloid gefunden (Typ E). Dieser Typ enthielt ausserdem ein wenig medulläres Amyloid in allen Fällen. Es ist die Frage, ob die verschiedenen Typen von Nierenlaesionen verschiedene Amyloidformen enthalten oder Varianten der selben Krankheit sind. Klinisch-pathologische Untersuchungen müssen diese Frage zu lösen versuchen.Medulläres Amyloid könnte eine subklinische Krankheit darstellen: Bei klinisch gesunden Schlachtrindern kam es in 2.7% von 1326 Tieren vor. Bei klinisch gesundem Milchvieh (einschliesslich Jungvieh) kam es in 2.6% von 378 Tieren vor, wobei mit steigendem Alter eine Zunahme gefunden wurde.Bei den medullären Amyloidablagerungen wurden mehrere amyloid-enthaltende Zelltypen gefunden. Das intrazelluläre Amyloid zeigte eine paralle Anordnung der Fibrillen und war lokalisiert in Lysosomen. Bei durch Injektion von Rinderamyloid bei Ratten induzierte Phagocytose zeigten sich die gleichen Vakuolen. Dieser Befund weist darauf hin dass Phagozytose auch bei Rindern eine Möglichkeit sein kann. In der Literatur wird paralleles intralysosomales Amyloid als örtlich entstanden erklärt. Schliesslich werden einige Untersuchungsrichtungen gezeigt, die möglicherweise zur pathogenese der Amyloidose führen könnten.

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Resume Après avoir résumé les points les plus importants de l'amyloidose relevés dans la littérature, l'auteur présente le résultat de ses observations. Il décrit l'aspect macroscopique et microscopique de l'amyloidose rénale des bovins. Dans la plupart des cas prédominent les dépôts médullaires (types A, B, C, D). Sur 1000 cas, il a trouvé 6 reins avec dépôts glomérulaires (type E), et partiellement médullaires.Une question se pose: les différents types de lésions rénales contiennent-ils diverses substances amyloides ou bien répresententils des variants d'un même affection. Des recherches clinico-histopathologiques devraient pouvoir apporter une réponse.L'amyloidose medullaire peut être une maladie sub-clinique. 2,7% de cas ont été décelés sur 1326 bovins de boucherie cliniquement sains et 2,6% sur 378 vaches laitières (y compris les jeunes) cliniquement saines.Les différents types de cellules contenant la substance amyloide existent dans les dépôts médullaires. La substance amyloide intracellulaire se présente en arrangements parallèles et est localisée dans les lysosomes. La phagocytose induits de la substance amyloide par les macrophages de Rat apparait sous la forme de vacuoles semblables, indiquant que la phagocytose peut se manifester dans les cellules bovines. Dans la littérature, la substance amyloid intra-lysosomale parailèle a été interpretée comme étant une formation de la substance amyloide de ces vacuoles.L'auteur suggere finalement quelques recherches pour élucider la pathogénie de l'amyloidose.

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Riassunto L'autore presenta il risultato delle sue osservazioni dopo aver riassunto i punti piu importanti dell'amiloidosi riportati in litteratura. Descrive sia l'aspetto macroscopico che microscopico dell'amiloidosi. Nella maggior parte dei casi precominano i depositi midollari (tipi A, B, C, D). Su 1.000 casi, ha trovato sei reni con depositi glomerulari (tipo E) e parzialmente midollari. Si pone la domanda se i diversi tipi di lesioni renali contangano sostanze amiloidi diverse, oppure se rappresentano dell varianti di una stessa affezione. Ricerche clinico-isopatologiche dovrebbero poter dare una risposta.L'amiloidosi midollare puo essere una malattia sub-clinica: il 2,7% dei casi e stato rilevato su 1.326 bovini da macello clinicamente sani, e su 378 vacche da latte clinicamente sane (ivi comprese le giovani), il tasso rilevato e del 2,6%. Nei depositi amilodali midollari si sono trovato vari tipi cellulari contenenti l'amiloide.L'amiloide intracellulare, che si localizza nei lisosomi, si presenta in strutture parallele. La fagocitosi della sostanza amiloide indotta dae microfaghi di ratto compare sotto forma di vacuoli simili, indicando che la fagocitosi puo manifestarsi nelle cellule bovine. In litterature, la sostanza amiloidale intralysomale parallela e stata interpretata come una formazione di sostanza amiloidale in questi vacuoli. Infine, l'autore suggerisce di fare alcune ricerche per chiarire la patogenesi dell'amiloidosi.

     

Serological prevalence of bovine babesiosis in Mali

Summary A serological survey of cattle in Mali was carried out to determine the prevalence of antibody activity toBabesia bovis andB. bigemina. It was found that the level ofB. bovis infection as indicated by antibody activities was too low to be of immediate concern. However, the serological prevalence ofB. bigemina was high and this may indicate a potential disease problem. It was also found that when zebu and N'Dama cattle grazed together the N'Dama were twice as likely to have positive titres toBabesia as were the zebus.

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Prevalencia Serologica De Babesiosis Bovina En Mali

Resumen Se llevó a cabo un análisis serológico en Malí, para detectar la prevalencia deBabesia bovis yBabesia bigemina. Se encontró, que la prevalencia serológica deB. bovis es baja para considerar la enfermedad una amenaza inmediata. Sinembargo, la prevalencia deB. bigemina fué alta, un problema potencial. Tambien se encontró, que cuando el ganado Cebu y N'Dama pastorea junto, el N'Dama presenta el doble de títulos serológicos deB. bigemina.

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Frequence Serologique De La Babesiose Bovine Au Mali

Résumé Une enquête sérologique sur le bétail au Mali a été effectuée pour déterminer la fréquence de l'activité des anticorps àBabesia bovis etB. bigemina. On a trouvé que le degré d'infection àB. bovis tel qu'indiqué par les réactions anticorps était trop faible pour constituer un souci immédiat. Cependant la fréquence sérologique deB. bigemina était élevée, ce qui peut indiquer un problème pathologique potentiel. On a aussi noté que lorsque les zébus et les N'Dama broûtaient ensemble, les N'Dama avaient deux fois plus de chance d'avoir des titres positifs àBabesia que ne l'avaient les zébus.

     

Purification and quantitative measurement of bovine serum amyloid-A

Bovine serum amyloid-A (b-SAA) was purified from a pool of acute phase serum using hydrophobic interaction chromatography and gel filtration. Serum was applied at a low salt concentration to a phenyl-sepharose column and SAA was eluted with a gradient of 0 to 6 M guanidine-HCI. Fractions containing SAA were pooled, concentrated and further purified by gel filtration on Superose-12. The concentration of SAA in bovine serum was quantified by an indirect ELISA using rabbit anti-human SAA and horseradish peroxidase conjugated donkey anti-rabbit IgG. Dilutions of an acute phase bovine serum sample were used as working standards. The SAA concentration of this standard was determined by comparison with purified b-SAA on SDS-polyacrylamide gel electrophoresis followed by densitometry at 590 nm. The assay detection limit was 3 μg ml⁻¹; the intra-assay coefficient of variation was 4 per cent and interassay coefficients of variation were 5·5 per cent and 7·2 per cent at 66 and 178 μg ml⁻¹ SAA, respectively. In calves experimentally infected with Pasteurella haemolytica type A1 the ELISA was able to detect a 10-fold increase of SAA within 24 hours of inoculation.     

Uterine infection with bovine herpesvirus type 4 in dairy cows

Diseases of the reproductive tract are a frequent problem in dairy herds. Herpesviruses are uterine pathogens also involved in other clinical diseases; for example, bovine herpesvirus type 4 BoHV‐4 induces abortion, enteritis, metritis, pneumonia and vaginitis, but it can also be detected in healthy cows. The role of BoHV‐4 in the development of clinical endometritis (CE) or subclinical endometritis (SE) has not clearly been described. Therefore, the objective of this study was to describe the prevalence of uterine BoHV‐4 infection and its relationship with clinical, bacteriological and cytological findings in dairy cows 20–30 days after calving. The experiment was performed as a completely randomized block design, with farm (n = 10) as blocking criterion and with cow (n = 397) as the experimental unit. Logistic regression models were used to assess the effect of BoHV‐4 infection on CE, SE and reproductive performance. Proportion of cows infected with BoHV‐4 was 5.8% (n = 23/397). BoHV‐4 was isolated in 11.0% (n = 12/109), 4.8% (n = 4/84) and 3.6% (n = 7/194) of cows diagnosed as CE, SE or healthy, respectively. A logistic model revealed that BoHV‐4 infection showed a tendency to increase the risk for CE (AOR = 2.17; p = .10) but significantly reduced both, the odds for artificial insemination within 80 days post‐partum (dpp) (AOR = 0.37; p = .035) and for pregnancy within 200 dpp (AOR = 0.13; p = .004). Furthermore, BoHV‐4 infection increased the chance for intrauterine infection with Trueperella pyogenes (AOR = 5.55; p < .001) and vice versa (AOR = 5.79, p < .001). In conclusion, BoHV‐4 infection is associated with reduced chances for insemination and pregnancy by 200 dpp and showed a trend to be associated with increased risk for CE. Furthermore, BoHV‐4 and Trueperella pyogenes infections are strongly related.     

Aspects of bovine herpesvirus 1 and bovine viral diarrhoea virus herd-level seroprevalence and vaccination in dairy and beef herds in Northern Ireland

Background

Infections with bovine herpesvirus 1 (BoHV-1) and bovine viral diarrhoea (BVD) virus cause diseases of cattle with a worldwide distribution. The primary objective of the present study was to describe aspects of herd-level BoHV-1 and BVDV seroprevalence (based on testing of pooled sera) and control on farms in Northern Ireland, including vaccine usage.An indirect antibody ELISA test (SVANOVA, Biotech AB, Uppsala, Sweden) was applied to serum pools which were constructed from serum samples taken for a cross-sectional study of a convenience sample of 500 Northern Irish dairy and beef cow herds in 2010, for which vaccination status was determined by telephone survey. The herd-level seroprevalence of BoHV-1 and BVDV in Northern Ireland was estimated in non-vaccinating herds and associations between possible risk factors (herd type and herd size (quartiles)) and herd-level prevalence were determined using chi-squared analysis.

Results

The herd-level seroprevalence (of BoHV-1 and BVDV) in non-vaccinating herds was 77.3% (95% CI: 73.6–80.9%) and 98.4% (95% CI: 97.3–99.5%) respectively in the cross-sectional study. A significant difference existed in BoHV-1 herd-level seroprevalence between dairy and beef herds (74.7% vs 86.5% respectively; p < 0.02) though not for BVDV seroprevalence (98.5% vs 98.3% respectively; p > 0.91). A significant association was found between herd size (quartiles) and herd-level classification for BoHV-1 herd-level seroprevalence based on cut-off percentage positivity (COPP) (p < 0.01) while no such association was found for BVDV (p = 0.22).15.5% and 23.8% of farmers used BoHV-1 and BVDV vaccines, respectively. BoHV-1 vaccine was used in 30% of dairy herds and in 11% of beef herds, while BVDV vaccine was used in 46% and 16% of dairy and beef herds, respectively.

Conclusions

The results from this study indicate that the true herd-level seroprevalences to bovine herpesvirus 1 and bovine virus diarrhoea virus in non-vaccinating herds in Northern Northern Ireland are 77.3% (95% CI: 73.6–80.9%) and 98.4% (95% CI: 97.3–99.5%), respectively. The present study will assist in guiding regional policy development and establish a baseline against which the progress of current and future control and eradication programmes can be measured.     

In vitro effects of lipopolysaccharides on bovine uterine contractility

Metritis is an important disorder in dairy cows during the early postpartum period. Myometrial contractility is a prerequisite for uterine involution; however, very scanty literature is available about the effect of metritis on this process and endocrine responsiveness. This study was aimed to evaluate the effect of inflammation on uterine contractility in vitro, and the inflammation was induced by incubating myometrial strips with lipopolysaccharides (LPS). Myometrial samples were collected from 17 healthy Holstein Friesian cows during caesarean section. Eight longitudinal strips from each cow were incubated in organ baths with LPS concentrations of 0 (LPS₀), 0.1 (LPS_0.1), 1 (LPS₁) and 10 µg/ml (LPS₁₀). Spontaneous contractility and contractility induced by increasing concentrations of oxytocin (10^–10 – 10^–7 mol/L) were recorded during nine 30-min intervals (T1 to T9). The minimum amplitude (minA), maximum amplitude (maxA), mean amplitude (meanA) and area under the curve (AUC) were calculated for each time interval. LPS had an effect (p ≤ .05) on maxA, meanA and AUC. In T1, myometrial strips incubated with LPS_0.1 and LPS₁ had higher (p ≤ .05) maxA, meanA and AUC than the strips incubated with LPS₀. In T9 without oxytocin, LPS₀ led to higher (p ≤ .05) maxA, meanA and AUC than LPS_0.1 and LPS₁. In T8 and T9 with oxytocin, LPS₁ had lower (p ≤ .05) maxA, meanA and AUC than the other LPS concentrations. Interestingly, the results show that LPS has a transient positive effect on myometrial contractility in vitro and that this effect is dependent on LPS concentration and duration of incubation.     

Ergot alkaloids reduce circulating serotonin in the bovine

Ergot alkaloids can interact with several serotonin (5-hydroxytryptamine [5-HT]) receptors provoking many physiological responses. However, it is unknown whether ergot alkaloid consumption influences 5-HT or its metabolites. Thus, two experiments were performed to evaluate the effect of ergot alkaloid feeding on 5-HT metabolism. In exp. 1, 12 Holstein steers (260 ± 3 kg body weight [BW]) were used in a completely randomized design. The treatments were the dietary concentration of ergovaline: 0, 0.862, and 1.282 mg/kg of diet. The steers were fed ad libitum, kept in light and temperature cycles mimicking the summer, and had blood sampled before and 15 d after receiving the treatments. The consumption of ergot alkaloids provoked a linear decrease (P = 0.004) in serum 5-HT. However, serum 5-hydroxytryptophan and 5-hydroxyindoleacetic acid did not change (P > 0.05) between treatments. In exp. 2, four ruminally cannulated Holstein steers (318 ± 3 kg BW) were used in a 4 × 4 Latin square design to examine the difference between seed sources on 5-HT metabolism. Treatments were: control—tall fescue seeds free of ergovaline, KY 32 seeds (L42-16-2K32); 5Way—endophyte-infected seeds, 5 way (L152-11-1739); KY31—endophyte-infected seeds, KY 31 (M164-16-SOS); and Millennium—endophyte-infected seeds, 3rd Millennium (L108-11-76). The endophyte-infected seed treatments were all adjusted to provide an ergovaline dosage of 15 μg/kg BW. The basal diet provided 1.5-fold the net energy requirement for maintenance. The seed treatments were dosed directly into the rumen before feeding. The experiment lasted 84 d and was divided into four periods. In each period, the steers received seeds for 7 d followed by a 14-d washout. Blood samples were collected on day 0 (baseline) and day 7 for evaluating the treatment response in each period. A 24 h urine collection was performed on day 7. Similar to exp. 1, serum 5-HT decreased (P = 0.008) with the consumption of all endophyte-infected seed treatments. However, there was no difference (P > 0.05) between the infected seeds. The urinary excretion of 5-hydroxyindoleacetic acid in the urine was not affected (P > 0.05) by the presence of ergot alkaloids. In conclusion, the consumption of ergot alkaloids decreases serum 5-HT with no difference between the source of endophyte-infected seeds in the bovine.     

Influence of seminal plasma during different stages of bovine sperm cryopreservation

This study aimed to evaluate the effect of seminal plasma on bovine sperm cryopreservation and to assess the integrity of plasma and acrosomal membranes, mitochondrial potential, remodelling of F-actin cytoskeleton and sperm chromatin fragmentation during the cooling, equilibrium and freezing/thawing stages. Six ejaculates collected from seven Nelore bulls (n = 42) were used in this study. Each ejaculate was divided into two aliquots (with seminal plasma = SP group; without seminal plasma = NSP group) and packed to a final concentration of 50 × 10⁶ sperm per straw. Statistical analyses were performed using SAS software (version 9.3), and p ≤ .05 was considered significant. A time effect was observed for all sperm characteristics (p < .05), except for chromatin fragmentation (p > .05). The presence of seminal plasma better preserved the acrosomal integrity (SP = 75.2% and NSP = 71.7%; p < .05) and also provided lower F-actin remodelling during cryopreservation process (SP = 29.9% and NSP = 32.4%; p < .05). Regarding to the cryopreservation stages, it was observed that cooling step induced higher remodelling of F-actin than the equilibrium and freezing/thawing stages (56.3%, 32.2% and 23.9%, respectively; p < .05). The equilibrium step had minor influence on overall sperm characteristics while the freezing/thawing stage was responsible for the highest percentage of damage in plasma membrane (−65.2%), acrosomal membrane (−34.0%) and mitochondrial potential (−48.1%). On the other hand, none of the cryopreservation stages affected chromatin integrity. It was concluded that the presence of seminal plasma provides increased acrosomal integrity and reduced remodelling of F-actin cytoskeleton. Higher F-actin remodelling is observed after the cooling step while the freezing/thawing step is most damaging to sperm membranes and mitochondrial potential during bovine sperm cryopreservation.