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[Analysis of POU1F1 gene polymorphisms in Qinchuan cattle and Chinese Holstein cattle].

PCR-RFLP was applied to analyze the polymorphisms of POU1F1 gene in 218 Qinchuan cattle (QQ) and Chinese Holstein cattle (HC). Results demonstrated Hinf I polymorphisms in the 451 bp PCR product in the two populations. The frequencies of alleles A/B in QQ and HC populations were 0.232/0.768 and 0.132/0.868, respectively. The frequencies of three genotypes AA, AB and BB were 0.030/0.403/0.567 and 0.007/0.251/0.742, respectively. Qinchuan cattle population was at Hardy-Weinberg equilibrium at this locus, but Chinese Holstein cattle population was not. The gene heterozygosity/effective allele gene number/Shannon information entropy/polymorphism information content of Qinchuan cattle and Chinese Holstein cattle populations were listed for 0.356/1.553/0.541/0.292 and 0.229/1.297/0.390/0.203, respectively. All indices were higher in the Qinchuan cattle population.

Animals↗

Identification of a doublet missense substitution in the bovine LRP4 gene as a candidate causal mutation for syndactyly in Holstein cattle.

Syndactyly in Holstein cattle is an autosomal recessive abnormality characterized by the fusion of the functional digits. This disorder has been previously mapped to the telomeric part of bovine chromosome 15. Here, we describe the fine-mapping of syndactyly in Holstein cattle to a 3.5-Mb critical interval using a comparative mapping approach and an extended pedigree generated by embryo transfer. We report genetic evidence for the exclusion of two genes previously suggested as candidates (EXT2 and ALX4) and describe the identification of a doublet mutation in complete linkage disequilibrium with syndactyly in one gene of the critical interval: LRP4. Finally, based on recent discoveries concerning the mouse mutants dan and mdig and a mouse knockout for Lrp4, we present solid evidence that the subsequent substitution in LRP4 exon 33 is a strong candidate causal mutation for syndactyly in Holstein cattle.

Amino Acid Sequence↗

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.

Animals↗

Relative efficiencies of wither height and body weight increase from birth until first calving in Holstein cattle.

Female Holstein calves from the Purina Research Center herd averaged 74 cm of height at withers and 40 kg of body weight (BW) at birth. At first calving (24 mo of age), calves averaged 138 cm of height at withers, 611 kg at precalving BW, and 547 kg at postcalving BW. Fifty percent of the total height increase occurred from birth to 6 mo of age, 25% occurred during 7 to 12 mo of age, and 25% occurred during 13 to 24 mo of age. Twenty-five percent of the total BW increase occurred from birth to 6 mo of age, 25% occurred from 6 to 12 mo of age, and the remaining 50% occurred from 13 to 24 mo of age. The increase in BW for 2-mo intervals, expressed as a proportion of the BW of the previous 2-mo interval, declined most rapidly during the first 6 mo, intermediately during the second 6 mo, and more slowly and progressively during the final 12 mo. Feed cost per unit of BW gain was lowest during the first 6 mo and then increased at a decreasing rate over the final 18 mo. The proportion of BW to height linearly increased over the 24-mo period. The increase in wither height as a proportion of the wither height during the previous period was greatest during the first 6 mo, intermediate during the second 6 mo, and lowest in the final 12 mo. Feed cost per unit of height increase was lowest in the first 6 mo, intermediate during the second 6 mo, and highest with a rapid increase during the final 12 mo. The increases in relative BW and wither height are the most rapid and cost efficient during the first 6 mo of life.

Aging↗

Serologically detected lymphocyte antigens in Holstein cattle.

Lymphocytes from 103 Holstein cattle were tested with 11 antilymphocytotoxic sera. Four of these sera were produced by whole blood immunization; these generally yielded lymphocytotoxicity against a large number of animals in each test panel. The majority of the sera were collected from parous cows which had never been immunized. Observations about this latter group of sera are (1) lymphocyte antibodies, may occur by alloimmunization in foetal-maternal interaction, (2) normal sera from non-immunized parous cows generally had a narrower specificity of antibodies than from parous cows immunized with blood from unrelated animals and in some cases these antisera may be monospecific, and (3) these sera from parous cows are easy to obtain without the need for laborious immunizations. Analysis of these lymphocytotoxic sera indicate several reagents were obtained which possessed different antibodies; these were useful in detecting polymorphism of cattle lymphocyte antigens. Using these 11 sera, an analysis of the reactions patterns among 103 Holsteins, including parent-offspring data resulted in the postulation of several alleles. These data suggest that cattle lymphocyte antigens are very polymorphic and inherited.

Alleles↗

Identification of vaccine candidate peptides in the NcSRS2 surface protein of Neospora caninum by using CD4+ cytotoxic T lymphocytes and gamma interferon-secreting T lymphocytes of infected holstein cattle.

Previously, our laboratory showed that Holstein cattle experimentally infected with Neospora caninum develop parasite-specific CD4+ cytotoxic T lymphocytes (CTL) that lyse infected, autologous target cells through a perforin-granzyme pathway. To identify specific parasite antigens inducing bovine CTL and helper T-lymphocyte responses for vaccine development against bovine neosporosis, the tachyzoite major surface proteins NcSAG1 and NcSRS2 were targeted. In whole tachyzoite antigen-expanded bovine T-lymphocyte lines, recombinant NcSRS2 induced potent memory CD4+- and CD8+-T-lymphocyte activation, as indicated by proliferation and gamma interferon (IFN-gamma) secretion, while recombinant NcSAG1 induced a minimal memory response. Subsequently, T-lymphocyte epitope-bearing peptides of NcSRS2 were mapped by using overlapping peptides covering the entire NcSRS2 sequence. Four experimentally infected cattle with six different major histocompatibility complex (MHC) class II haplotypes were the source of immune cells used to identify NcSRS2 peptides presented by Holstein MHC haplotypes. NcSRS2 peptides were mapped by using IFN-gamma secretion by rNcSRS2-stimulated, short-term T-lymphocyte cell lines, IFN-gamma enzyme-linked immunospot (ELISPOT) assay with peripheral blood mononuclear cells, and 51Cr release cytotoxicity assay of rNcSRS2-stimulated effector cells. Four N. caninum-infected Holstein cattle developed NcSRS2 peptide-specific T lymphocytes detected ex vivo in peripheral blood by IFN-gamma ELISPOT and in vitro by measuring T-lymphocyte IFN-gamma production and cytotoxicity. An immunodominant region of NcSRS2 spanning amino acids 133 to 155 was recognized by CD4+ T lymphocytes from the four cattle. These findings support investigation of subunit N. caninum vaccines incorporating NcSRS2 gene sequences or peptides for induction of NcSRS2 peptide-specific CTL and IFN-gamma-secreting T lymphocytes in cattle with varied MHC genotypes.

Amino Acid Sequence↗

[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.

Animals↗

Sequence variation of bovine prion protein gene in Japanese cattle (Holstein and Japanese Black).

To assess relationships between nucleotide polymorphisms of the prion protein (PRNP) gene and susceptibility to bovine spongiform encephalopathy (BSE), we investigated polymorphisms in the open reading frame (ORF) and 2 upper regions of the PRNP gene from 2 Japanese cattle breeds: 863 healthy Holstein cattle, 6 BSE-affected Holstein cattle, and 186 healthy Japanese Black (JB) cattle. In the ORF, we found single-nucleotide polymorphisms (SNPs) at nucleotide positions 234 and 576 and found 5 or 6 copies of the octapeptide repeat, but we did not find any amino acid substitutions. In the upper region, we examined 2 sites of insertion/deletion (indel) polymorphisms: a 23-bp indel in the upper region of exon 1, and a 12-bp indel in the putative promoter region of intron 1. A previous report suggests that the 23-bp indel polymorphism is associated with susceptibility to BSE, but we did not find a difference in allele frequency between healthy and BSE-affected Holstein cattle. There were differences in allele frequency between healthy Holstein and JB cattle at the 23- and 12-bp indels and at the SNPs at nucleotide positions 234 and 576, but there was no difference in allele frequency of the octapeptide repeat. We identified a unique PRNP gene lacking a 288-bp segment (96 amino acids) in DNA samples stocked in our laboratory, but this deletion was not found in any of the 1049 cattle examined in the present study. The present results provide data about variations and distribution of the bovine PRNP gene.

Animals↗

Analysis and frequency of bovine lymphocyte antigen (BoLA-DRB3) alleles in Iranian Holstein cattle.

The bovine lymphocyte antigen (BoLA-DRB3) gene encodes cell surface glycoproteins that initiate immune response by presenting processed antigenic peptides to CD4 T helper cells. DRB3 is the most polymorphic bovine MHC class II gene which encodes the peptide-binding groove. DRB3 gene has been extensively evaluated as a candidate marker for association with various bovine diseases and immunological traits. This study describes genetic variability in the BoLA-DRB3 in Iranian Holstein cattle. This is the first study of the DNA polymorphism of the BoLA-DRB3 gene in Iranian Holstein cattle. Hemi-nested PCR-RFLP method is used for identification the frequency of BoLA-DRB3 alleles. The BoLA-DRB3 locus is highly polymorphic in the studied herd (26 alleles). Almost 67% of the alleles were accounted for four alleles (BoLA-DRB3.2*8, *24, *11 and *16) in Iranian Holstein cattle. The DRB3.2*8 allele frequency (26.6%) was higher than the others. The frequencies of the DRB3.2*54, *37, *36, *28, *25, *14, *13, *10, *1 alleles were lower than 1%. Significant distinctions have been found between Iranian Holstein cattle and other cattle breeds studied. In Iranian Holstein cattle the alleles (BoLA-DRB3.2*22, *2 and *16) associated with a lower risk of cystic ovarian disease in Holstein cattle are found. The alleles associated with the resistance to mastitis and to bovine leukemia virus infection BoLA-DRB3.2*11 and *23 are detected with the frequencies 10.4% and 4.4%, respectively. Thus in the Iranian Holstein cows studied are found alleles which are associated with resistance to various diseases. The method of DNA-typing of animals can be used in agricultural practice for BoLA-DRB3 allele genotyping of cattle in order to reduce spreading of alleles providing susceptibility to mastitis or leukemia in cattle herds.

Animals↗

Hydatidiform moles in Holstein cattle.

Two hydatidiform moles from Holstein cattle are reported. One was observed after the birth of a healthy calf and the second following abortion of a fetus with osteopetrosis.

Animals↗

Polymorphisms of the prion protein gene (PRNP) in Hanwoo (Bos taurus coreanae) and Holstein cattle.

Polymorphisms in the prion protein gene (PRNP) in humans and sheep correlate with susceptibility to transmissible spongiform encephalopathies (TSEs). Bovine spongiform encephalopathy (BSE) has been reported in British and Japanese cattle; it has occurred thus far in Holstein cattle. BSE in Hanwoo (Bos taurus coreanae) cattle has not been diagnosed up to now. To characterize the bovine PRNP polymorphisms in Korean cattle, we analyzed the open reading frame (ORF) of PRNP in 120 Hanwoo (beef) cattle and 53 Holstein (dairy) cattle. Three polymorphisms were found, the third position of codon 78 (G-->A), the third position of codon 192 (C-->T), and the deletion of a single octa-repeat. An analysis of codon 78 revealed no difference in the genotype (P = 0.2026) or allele (P = 0.7180) frequencies between Hanwoo and Holstein animals. However, there were significant differences in the genotype (P < 0.0001) and allele (P < 0.0001) frequencies at PRNP codon 192 between Hanwoo and Holstein animals. The rate of Holstein animals with deletion of a single octa-repeat was 91.5% undeleted homozygotes, 8.5% heterozygotes (with R3 deletion), and 0% deleted homozygotes. However, none of the 120 Hanwoo animals had any octa-repeat deletions. The genotype (P < 0.0001) and allele (P < 0.0001) frequencies of a single octa-repeat-deletion were also significantly different between Hanwoo and Holstein animals.

Alleles↗

Effect of gestational mastectomy on postpartum gonadotropin releasing hormone and thyrotropin releasing hormone-induced luteinizing hormone and prolactin response in first lactation Holstein cattle.

First lactation Holstein cows were divided into two treatment groups to evaluate thyrotropin releasing hormone (TRH, 0.25 microgram/kg body weight) and gonadotropin releasing hormone (GnRH; 200 micrograms) induced secretion of prolactin (PRL) and luteinizing hormone (LH) on days 7 and 16 postpartum. Disregarding treatment, LH response was greater (p less than 0.01) on day 16 than day 7 postpartum (7.5 +/- 0.3 ng/ml on day 7 vs 10.2 +/- 0.3 ng/ml serum on day 16). Mastectomized cattle had similar time for initiation of LH increase, but peak concentrations were achieved later. Peak PRL concentrations were reached 12 to 15 min after injection and returned to baseline within 2.5 h in both groups. However, intact cows had higher (p less than 0.01) mean serum PRL than the mastectomized cows for 1 h following injection. Peak PRL concentration was 83.3 +/- 17.6 ng/ml for mastectomized cows vs 128.0 +/- 24.7 ng/ml for intact cows. It appears that udder removal allows for greater pituitary responsiveness to GnRH but diminishes PRL response to TRH suggesting the mammary gland differentially affects pituitary secretion of LH and PRL.

Animals↗

Identification and prevalence of a genetic defect that causes leukocyte adhesion deficiency in Holstein cattle.

Two point mutations were identified within the gene encoding bovine CD18 in a Holstein calf afflicted with leukocyte adhesion deficiency (LAD). One mutation causes an aspartic acid to glycine substitution at amino acid 128 (D128G) in the highly conserved extracellular region of this adhesion glycoprotein, a region where several mutations have been found to cause human LAD. The other mutation is silent. Twenty calves with clinical symptoms of LAD were tested, and all were homozygous for the D128G allele. In addition, two calves homozygous for the D128G allele were identified during widespread DNA testing, and both were subsequently found to exhibit symptoms of LAD. The carrier frequency for the D128G allele among Holstein cattle in the United States is approximately 15% among bulls and 6% among cows. This mutation is also prevalent among Holstein cattle throughout the world, placing this disorder among the most common genetic diseases known in animal agriculture. All cattle with the mutant allele are related to one bull, who through the use of artificial insemination sired many calves in the 1950s and 1960s. The organization of the dairy industry and the diagnostic test described herein will enable nearly complete eradication of bovine LAD within 1 year. These results also demonstrate that bovine LAD is genetically homologous and phenotypically similar to human LAD, thus providing a useful animal model for studies of LAD and beta 2 integrin function.

Alleles↗

Detection of bovine leukocyte adhesion deficiency (BLAD) in Turkish native and Holstein cattle.

The purpose of this work was to study whether the bovine leukocyte adhesion deficiency (BLAD) allele is present in native cattle breeds and the Holstein breed in Turkey. Blood samples were obtained from 120 Holstein, 20 Brown Swiss, 20 Anatolian Black, 20 Turkish Grey, 20 South Anatolian Red and 20 East Anatolian Red cattle. The isolated DNA materials were multiplied in PCR using the primer developed by Kriegesmann et al. (1997). In order to determine the area of mutation in PCR products, the PCR products were digested with TaqI endonuclease enzyme. The resulting fragments were analysed on 2% agarose gel for the absence of a TaqI restriction site. It was found that two of the Holstein cattle (a bull and a cow) were heterozygote BLAD carriers. There was no homozygote BLAD animal. The BLAD allele was not found in the other breeds used in the study. The mutant BLAD allele frequency in the 120 Holstein cattle calculations was 0.0084.

Alleles↗

The association between reproductive performance and milk yield in Chilean Holstein cattle.

The objective of this study was to evaluate the relationship between reproductive performance and milk yield in central-southern Chilean Holstein cattle that calved from 1990 to 2003. The analysis included 150,457 lactations obtained from a certified recording system. Reproductive indexes included in the study were calving interval (CI, d), calving to first service interval (CFSI, d), calving to conception interval (CCI, d), services per conception (SC), and conception rate at first service (CRFS). Survival analysis for the risk of pregnancy was also conducted. Models for reproductive indexes were significant and included, as independent variables, year and season of parturition, parity, length of dry period, milk and fat production standardized to 305 d, herd size, and herd. In 1990 and 2003, respectively, means +/- SEM for CI were 399 +/- 1.6 and 415 +/- 1.1 d; for CFSI were 85 +/- 0.6 and 97 +/- 0.6 d; for CCI were 124 +/- 1.3 d and 137 +/- 1 d; and for SC were 1.6 +/- 0.02 and 1.7 +/- 0.01. For every 100 kg of 305-d standardized milk yield, the CCI increased by 0.6 d and CRFS decreased by 0.9%. Association between milk yield and the risk of pregnancy was almost zero when a Cox proportional regression model was conducted (hazard ratio = 1.005; 95% confidence interval = 1.002 to 1.008). We conclude that CCI has increased over time and is related negatively to the increase in milk yield experienced by central-southern Chilean Holstein cattle during the last 15 yr. Nevertheless, risk of pregnancy was not explained by the individual level of standardized 305-d milk yield of cows studied.

Animals↗

The morphology of the hypoglossal dorsal root and its ganglia in Holstein cattle.

This study was carried out to describe morphologically and morphometrically the hypoglossal dorsal root and its ganglia, using heads of 20 adult Holstein cattle of both sexes. The dorsal root of the hypoglossal nerve (XII) has always an evident dorsal root ganglion (DRG) including the accessory in 30% (6 of heads) or satellite ganglia in 10% (2 of heads). There were statistically significant differences between the lengths (right: 8.64+/-0.22 mm, left: 7.91+/-0.31 mm, p<0.01) of peripheral roots of the DRG in male. Statistical significance existed in the lengths (right: 7.01+/-0.43, left: 8.27+/-0.47, p<0.05) of the central roots of the DRG in male. Since the histological findings showed that each ganglion placed on the dorsal root of the XII had the general feature of a spinal ganglion, it can be said that the XII has the intracranial peripheral (sensory) fibers and carry directly sensory input to the brain stem. In conclusion, the present study is the first to reveal the morphology of the hypoglossal dorsal root with ganglia in Holstein cattle.

Animals↗

Glucose metabolism in vivo in crossbred Holstein cattle feeding on different types of roughage during late pregnancy and early lactation.

An experiment was carried out to study the glucose kinetics of crossbred Holstein cattle feeding on either hay or 5% urea treated rice straw during late pregnancy (21 days prepartum) and early lactation (30 days postpartum). In all 16 pregnant heifers (23-25 months of age) were selected for the experiments, including eight animals of two breed types, Holstein Friesian x Red Sindhi (50:50 = 50% HF) and Holstein Friesian x Red Sindhi (87.5:12.5 = 87.5% HF). They were divided into four groups of four animals each. Animals from the same breed type in each group were fed with either rice straw treated with 5% urea or pangola hay (Digitaria decumbens) as the source of roughage throughout the experiments. The glucose turnover rate in both types of crossbred Holstein cattle was determined using a continuous infusion of [U-14C] and 3-[3H]glucose during late pregnancy and early lactation. Total glucose entry and utilization rates increased significantly during lactation for all groups. Recycling of [C]glucose was, approximately 20% in both crossbred cattle fed either hay or urea treated rice straw and was unaffected by the stage of late pregnancy or early lactation. Comparing 50 and 87.5% HF animals, arterial plasma glucose concentrations were slightly higher during pregnant periods but significantly higher in lactating periods in 50% HF animals. The ratio of specific radioactivity of arterial blood bicarbonate relative to that of arterial blood [14C]glucose in the lactating period, significantly decreased in 50% HF animals fed either urea treated rice straw or hay. An increase in udder blood flow during early lactation was significantly higher in 87.5% HF animals than in 50% HF animals. The uptake, arteriovenous differences and extraction ratio for glucose across the udder, significantly increased in the lactating period for all crossbred animals. Glucose uptake by the udder of 87.5% HF animals accounted for 65% of the total glucose turnover rate compared to a value of 46% in the lactating 50% HF animals. It can be concluded that both crossbred cattle fed either urea treated rice straw or hay exhibit the same body glucose turnover rate. The 87.5% HF animal has the genetic potential for a high milk yield and has high body and udder glucose metabolisms compared with 50% HF animals.

Animal Feed↗

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.

Abortion, Veterinary↗