Echo-guided studies on portal and hepatic blood in cattle

The objectives of the present study were (1) to establish and maintain catheters non-surgically in the portal and hepatic veins of cattle for long-term monitoring of postprandially absorbed metabolites, and (2) to assess the safety of the technique used. Aspiration and catheterization were not performed if there was risk of bleeding, as determined by hemostatic data and enlargement of portal and hepatic veins. In five Holstein cattle, before and 60-min after feeding, percutaneous ultrasound-guided aspiration of portal and hepatic blood, together with jugular blood, was performed utilizing 17G x 17.5 cm needles. Concentration of ammonia nitrogen tended to be higher after feeding, but that of urea nitrogen and glucose were not significantly changed. In five other Holstein cows liver veins were catheterized with 14 x 16.5 cm guide-needles attached with 50 cm polyethylene tubes and blood samples were collected from portal, hepatic and jugular veins at 30-min intervals for 4.5 hr. The concentration of plasma acetate and 3-hydroxybutyrate (3-HB) were significantly increased after feeding, while that of lactate was not changed. Catheters in the portal and hepatic veins enabled the collection of blood samples up to 15 days (mean, 7 days). After a 7-day observation period, five cows were euthanised and necropsied. Changes in the intercostal musculature, peritoneum and liver were negligible. During the observation period, clinical profiles, aspartate aminotransferase, alanine aminotransferase and total and differential WBC counts were within normal ranges.     

A hereditary disposition for bovine peripheral nerve sheath tumors in Danish Holstein cattle

Background

Peripheral nerve sheath tumors (PNSTs) are frequently found in Danish cattle at slaughter. Bovine PNSTs share several gross and histopathological characteristics with the PNSTs in humans with heritable neurofibromatosis syndromes. The aim of the present study was to investigate a possible hereditary disposition to PNSTs in dairy cattle by statistical analysis performed on data from 567 cattle with PNSTs. Furthermore, a preliminary genome-wide association study (GWAS) was performed on DNA isolated from 28 affected and 28 non-affected Holstein cows to identify loci in the bovine genome involved in the development of PNSTs.

Results

PNSTs were significantly more common in the Danish Holstein breed than in other breeds with 0.49% of Danish Holsteins slaughtered during an eight-year-period having PNSTs. PNSTs also occurred significantly more frequently in the offspring of some specific Holstein sires. Examination of three generation pedigrees showed that these sires were genetically related through a widely used US Holstein sire. The PNSTs included in GWAS were histologically classified as neurofibroma-schwannoma (43%), schwannoma (36%) and neurofibroma (21%) and derived from Holstein cows with multiple PNSTs. A single SNP on chromosome 27 reached genome-wide significance.

Conclusions

Gross and histological characteristics of bovine PNSTs are comparable to PNSTs in humans (schwannomatosis). Danish Holsteins are genetically disposed to develop PNSTs but the examined materials are insufficient to allow determination of the mode of inheritance.     

Brachyury in German Holstein cattle

In 11 German Holstein cattle with black and white or red and white coat colour short tails could be observed. All affected animals were born in different farms. A clinical examination could not reveal further congenital anomalies of other organs. The parents of the affected animals showed no abnormalities of the tail. The pedigree data showed that all short tailed animals were related with each other and this fact provides evidence that brachyury is an inherited congenital anomaly. The coefficients of relationships were between 0.20 and 25%. The pedigree indicated a monogenic or oligogenic autosomal recessive inheritance. Environmental factors causing short tails in these animals were not obvious.     

Hepatic steroid inactivating enzymes,hepatic portal blood flow and corpus luteum blood perfusion in cattle

Production from the corpus luteum (CL) and/or hepatic steroid inactivation impacts peripheral concentrations of P4, which can alter reproductive performance. Our primary objective was to examine hepatic steroid inactivating enzymes, portal blood flow, and luteal blood perfusion at 10 days post‐insemination in pregnant versus non‐pregnant beef and dairy cows. Twenty early lactation Holstein cows and 20 lactating commercial beef cows were utilized for this study. At day 10 post‐insemination, hepatic portal blood flow and CL blood perfusion were measured via Doppler ultrasonography. Liver biopsies were collected and frozen for later determination of cytochrome P450 1A (CYP1A), 2C (CYP2C), 3A (CYP3A), uridine diphosphate‐glucuronosyltransferase (UGT) and aldo‐keto reductase 1C (AKR1C) activities. Pregnancy was determined at day 30 post‐insemination and treatment groups were retrospectively assigned as pregnant or non‐pregnant. Data were analyzed using the mixed procedure of SAS. Steroid metabolizing enzyme activity was not different (p > .10) between pregnant versus non‐pregnant beef or dairy cows. Hepatic portal blood flow tended (p < .10) to be increased in pregnant versus non‐pregnant dairy cows. Luteal blood perfusion was increased (p < .05) in pregnant versus non‐pregnant dairy cows. Pregnant dairy cows appear to have an increased rate of hepatic clearance of P4 in combination with increased synthesis from the CL. This could account for the lack of difference in peripheral P4 concentrations between pregnant and non‐pregnant dairy cows. This study highlights the relevance of further investigation into steroid secretion and inactivation and their impact on the maintenance of pregnancy in cattle.     

Comparison of the peripheral blood leukocyte population between Japanese Black and Holstein calves

Japanese black (JB) calves have greater susceptibility to infectious diseases compared to Holstein (Hol) calves. In order to clarify the differences in cellular immune status between JB and Hol calves, the leukocyte population and lymphocyte proliferative ability were analyzed. In total 200 healthy calves, 1 day to 14 weeks of age, were examined: 105 JB and 95 Hol calves. Lower numbers in peripheral blood and percentage in peripheral blood mononuclear cells of CD3⁺TcR1‐N12⁺ T cells and major histocompatibility complex class‐II⁺CD14^‐ B cells were observed in the JB compared to the Hol. The percentage of TcR1‐N12⁺CD25⁺ T cell in the JB was significantly lower than that of the Hol at 4–6, and 8–10 weeks. Interleukin (IL)‐2 sensitivity in the JB was lower than that in the Hol, and significant differences were observed in age groups of 6–8 weeks and 10–14 weeks. These findings indicated that the lower numbers of γδ T cells and B cells in the JB compared to the Hol might be associated with the specificity of the immune systems in JB calves.     

Changes in mononuclear peripheral blood cells in cattle with foot-and-mouth disease

The percentages and absolute numbers of mononuclear peripheral blood cells (MNC) were studied in vaccinated (Vac) and non-vaccinated (control) cattle, challenged with foot-and-mouth disease virus (FMDV). All Vac cattle but none of the controls resisted challenge. Cell populations were studied immediately before and one week after challenge, by direct and indirect immunofluorescence, using polyclonal and monoclonal antibodies against different bovine markers. Total B-lymphocytes, as assessed with polyclonal anti-IgG(H+L) antisera, as well as total mononuclear cells, were normal before and after infection, and did not change in Vac or control groups. Before challenge Vac cattle had higher numbers of IL A-29⁺ (a putative marker for null cells or, alternatively, for γδ T-cells) than control cattle. After challenge, in control cattle, the number of total T-cells, BoT4-bearing (helper) T-cells and BoT8-bearing (cytotoxic/suppressor) T-cells were decreased, while IgM-bearing B-lymphocytes, as well as monocyte/macrophage cells were increased. The number of IL-A29-bearing T-cells did not change after infection in either group. After challenge, Vac cattle also showed increased numbers of IgM-bearing B-lymphocytes and monocyte/macrophage cells, whereas T-subpopulations did not change significantly.     

Neospora caninum serostatus and culling of Holstein cattle

OBJECTIVE: To determine whether time until culling or risk of culling was associated with Neospora caninum serostatus among Holstein cattle in dairy herds in Ontario. DESIGN: Retrospective cohort study. ANIMALS: 3,412 cows in 56 herds. PROCEDURE: Blood samples were collected, and serum was tested for antibodies against N. caninum. Information on cows that were culled was collected during the 1- to 2-year period that producers were unaware of serostatus of individual cows in their herds. RESULTS: Herd prevalence of N. caninum-seropositive cows ranged from 0 to 68.3% (median, 7.0%). During the time of the study, 184 of 359 (51.3%) N. caninum-seropositive cows were culled, compared with 1,388 of 3,053 (45.5%) seronegative cows. Mean time from blood sample collection to culling for seronegative cows (289 days; 95% confidence interval, 280 to 299 days) was not significantly different from mean time for seropositive cows (296 days; 95% confidence interval, 269 to 323 days). Survival analysis indicated that N. caninum serostatus was not associated with time to culling or risk of culling. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that N. caninum serostatus of Holstein cows in Ontario was not significantly associated with either time to culling or risk of culling. Thus, N. caninum serostatus alone should not be used to determine whether cows should be culled.     

夏季皖东牛与荷斯坦牛皮肤汗腺及其散热调节特征

为了探究夏季高温时皖东牛与荷斯坦牛皮肤组织形态和皮肤散热调节特征的差异,在30℃以上的夏日,选择皖东牛和荷斯坦牛母牛各4头,屠宰后采集颈部、肩部、背部、腹部、臀部和腿部6个部位的皮肤样本,通过HE染色切片分析皮肤的汗腺(SWG)与皮脂腺(SEG)的分布与形态,测定皮肤组织乙酰胆碱(ACh)浓度及乙酰胆碱转移酶(AChT)和酯酶(AChE)活力,采用RT-qPCR和Western blot检测皮肤组织乙酰胆碱M3受体(ACh M3R)、一氧化氮合酶(eNOs)及水通道蛋白5(AQP5)mRNA和蛋白的表达。结果:部位间比较,皖东牛与荷斯坦牛表皮厚度均为背部最厚,颈部最薄(P<0.05)。品种间比较,皖东牛平均皮肤厚度较高,平均表皮厚度较薄(P<0.05);汗腺密度与皮脂腺密度品种间无显著差异(P>0.05);皖东牛汗腺和皮脂腺的平均长度、直径、深度、体积均显著增加(P<0.05)。肩部、背部和臀部皮肤对比:夏季皖东牛和荷斯坦牛皮肤组织ACh浓度品种间无显著差异(P>0.05),部位间以臀部最高,肩部次之,背部最低(P<0.05)。ACh M3R和AQP...     

Genetic analysis of syndactyly in German Holstein cattle

Congenital syndactyly with a variable number of affected feet was observed in eight black and white German Holstein calves. Analysis of the pedigree data revealed that all affected individuals could be traced back to a single founder. The pedigree was consistent with monogenic autosomal recessive inheritance and variable expressivity. Bovine syndactyly or "mulefoot" has been previously shown to map on the telomeric end of bovine chromosome 15 and we performed PCR genotyping of microsatellite markers spanning 27 cM of this chromosomal region to test the new cases for genetic linkage with the phenotype. The haplotype segregation confirmed the suggested inheritance pattern of the mulefoot mutation in this family and markers RM004, BM848 and BMS820 showed significant linkage to the phenotype. The results confirmed the chromosomal location of the mulefoot gene in this pedigree. Furthermore the study demonstrated that although marker testing has been available for nearly a decade the use of mulefoot carriers in cattle breeding remains uncontrolled. The presented family provides a resource for positional cloning of the causative mutation.     

Effects of dopamine, norepinephrine and serotonin on secretion of luteinizing hormone, follicle-stimulating hormone and prolactin in ovariectomized, pituitary stalk-transected ewes

Two experiments were conducted in ovariectomized, pituitary stalk-transected ewes to determine if dopamine (DA), norepinephrine (NE) or serotonin (5-HT) alter secretion of luteinizing hormone (LH), follicle-stimulating hormone (FSH) and prolactin (PRL). In experiment 1, ewes were infused (iv) with saline (control), DA (66 micrograms/kg/min), NE (6.6 micrograms/kg/min) or 5-HT (6.6 micrograms/kg/min). Treatments did not alter pulse frequency, but 5-HT increased (P less than .05) amplitude of pulses of LH and mean concentrations of LH, DA and NE were without effect on basal secretion of LH. DA but not NE or 5-HT decreased (P less than .05) the release of LH in response to gonadotropin hormone-releasing hormone (GnRH, 25 micrograms, im). Concentrations of FSH were not affected by treatments. Secretion of PRL was reduced (P less than .05) by treatment with DA and NE but not 5-HT. Each amine reduced (P less than .05) the release of PRL in response to thyrotropin-releasing hormone (TRH; 3 micrograms, im). In experiment 2, ewes were given DA at doses of 0, 0.66, 6.6 or 66.0 micrograms/kg/min, iv. No dose altered basal LH, but each dose reduced (P less than .05) basal and TRH-induced release of PRL. Key findings from these studies include direct pituitary action for: (1) 5-HT enhanced basal secretion of LH, (2) suppression of GnRH-induced secretion of LH by DA. (3) DA and NE inhibition of PRL secretion, and (4) DA, NE and 5-HT inhibition of release of PRL in response to TRH.     

围产科尔沁牛外周淋巴细胞HSP70的表达

选择经同期发情处理的妊娠科尔沁牛5头(2³胎),于分娩-14、-6、-2、0、2、6、14d颈静脉采血,分离淋巴细胞,采用RT-PCR、Western blotting法分别检测淋巴细胞的HSP70mRNA与蛋白表达。结果显示,各受试牛的HSP70mRNA与蛋白表达变化趋势基本一致,即从-14d开始逐渐下调,0d最低,继而再逐渐上调。结果表明,HSP70作为淋巴细胞内重要伴侣分子,随着分娩的启动和发生其表达抑制显著增强,但在分娩后被迅速解除,其表达逐渐上调。     

围产科尔沁牛外周淋巴细胞HSP70的表达

选择经同期发情处理的妊娠科尔沁牛5头（2～3胎）,于分娩-14、-6、-2、0、2、6、14d颈静脉采血,分离淋巴细胞,采用RT～PCR、Westernblotting法分别检测淋巴细胞的HSP70mRNA与蛋白表达。结果显示,各受试牛的HSP70mRNA与蛋白表达变化趋势基本-致,即从-14d开始逐渐下调,0d最低,继而再逐渐上调。结果表明,HSP70作为淋巴细胞内重要伴侣分子,随着分娩的启动和发生其表达抑制显著增强,但在分娩后被迅速解除,其表达逐渐上调。     

Bovine leukocyte adhesion deficiency in Holstein cattle.

Two Holstein heifers with persistent and recurrent infections including ulcerative gingivitis, periodontitis, pneumonia, loss of teeth and stunted growth associated with marked neutrophilia were evaluated clinically and for neutrophil function, CD18 expression on neutrophils and CD18 genotype analysis by DNA-polymerase chain reaction (PCR) test. Adherence to nylon fibers and phagocytic activity of neutrophils from affected animals were significantly (p < 0.05) impaired as compared with those of controls. Neutrophils from affected heifers had decreased chemiluminescent (CL) responses when stimulated with opsonized zymosan, compared with those of controls. In contrast, neutrophils from affected heifers produced increased CL responses when stimulated with latex beads and phorbol myristate acetate compared with those of controls. The clinical findings, functional leukocyte abnormalities, deficiency in expression of CD18 on neutrophils, and the D128G mutation detected by DNA-PCR testing of affected heifers demonstrated that these heifers have bovine leukocyte adhesion deficiency (BLAD). Although both animals were confirmed to be homozygotes for BLAD by DNA-PCR test, they had differences in clinical, hematological and neutrophil functional characteristics.     

Udder edema: biochemical studies in Holstein cattle

Udder edema was studied in 9 affected and 3 control Holstein cows ranging in age from 2.5 to 8 years. A complete physical examination was done in conjunction with serum analysis for total protein, albumin, globulin, sodium, potassium, chloride, calcium, and inorganic phosphorus 2 weeks before parturition, at parturition and 2 weeks following parturition. Mammary interstitial fluid samples also were obtained from cows with udder edema and analyzed for total protein, albumin, globulin, sodium, potassium, chloride, calcium, and inorganic phosphorus. Clinical signs of udder edema appeared as early as 7 days prepartum and persisted for as long as 9 days postpartum. There were no significant differences in serum constituents between affected and control cows at the same stage of pregnancy or lactation. There was a significant difference (P less than or equal to 0.05) between serum and mammary interstitial fluid in the total protein, albumin, globulin and calcium concentrations.     

Neospora caninum serostatus and milk production of Holstein cattle

OBJECTIVE: To determine whether Neospora caninum serostatus was associated with milk production among Holstein cattle in Ontario. DESIGN: Case-control study and cross-sectional observational study. ANIMALS: 3,702 Holstein cows in 83 herds (case-control study) and 3,162 Holstein cows in 57 herds. PROCEDURE: Herds in the case-control study were grouped on the basis of N. caninum abortion status. Herds in the observational study were considered representative of Ontario dairy herds. The N. caninum serostatus of individual cows was determined with a kinetic ELISA. Milk production was modeled to compare seropositive with seronegative animals while controlling for parity, days since parturition, and herd clustering. RESULTS: In the case-control study, 305-day milk production of seropositive cows was significantly less than milk production of seronegative cows in herds with abortions attributable to N. caninum infection and in herds with abortions attributable to pathogens other than N. caninum, but not in herds without abortion problems. In the observational study, 305-day milk production for seropositive cows was not significantly different from milk production of seronegative cows. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that the association between N. caninum serostatus and milk production in Ontario Holstein dairy cattle may depend on abortion status of the herd. In herds with abortion problems, regardless of cause, N. caninum-seropositive cattle produced less milk, whereas in herds without abortion problems, N. caninum-seropositive cattle produced the same amount of milk as seronegative cattle.