Comparison of cytogenetics and polymerase chain reaction based detection of the amelogenin gene polymorphism for the diagnosis of freemartinism in cattle.

A polymerase chain reaction (PCR) assay which detects a sex-based polymorphism in the bovine amelogenin locus was modified and compared to conventional cytogenetic analysis for diagnosis of freemartinism (XX/XY chimerism) in cattle. The PCR assay is more sensitive than cytogenetic analysis for detection of XY cells, with the limit of detection of the assay falling between 0.2% and 1% XY cells. Seventy-three heifer blood samples submitted for evaluation of freemartinism to the University of Minnesota Diagnostic Laboratory were tested using both cytogenetic and PCR techniques. Poor-quality samples precluded successful lymphocyte culture and recovery of mitotic nuclei for cytogenetic evaluation in 17 cases (23%). Two of these samples (2.7%) also failed to amplify with PCR. There was 100% agreement in the results from the 56 samples that were suitable for testing using both techniques. This PCR-based assay provides an alternative to the more laborious cytogenetic evaluation for diagnosis of freemartinism.     

XX/XY chimerism and freemartinism in a female llama co-twin to a male.

A 20-month-old female llama was examined because at the time of mating, the male llama was apparently unable to achieve intromission. The female llama had been born co-twin to a male. On physical examination, the vaginal vestibule appeared to end in a blind pouch, and the uterus, cervix, and ovaries could not be identified during transrectal palpation or ultrasonography. Karyotyping was done, and 43% of blood lymphocytes had 2 X chromosomes, and 57% had 1 X and 1 Y chromosome. All skin fibroblasts had 2 X chromosomes. A diagnosis of freemartinism and XX/XY chimerism was made. Because conception of twins may be more common in llamas than birth of twins, it is possible that freemartinism could develop in singleton females, if, for instance, a male twin was conceived and died after the placentas had anastomosed. More widespread use of karyotyping in llamas with congenital defects of the reproductive tract will help to define the incidence of freemartinism.     

奶牛双胎的发生及其对奶业生产的影响

奶牛在一个发情周期内如果同时排出两个成熟卵子，经过受精和发育，就可能出现双胎现象。奶牛双胎的发生与遗传、产奶量、胎次、受孕年份和季节等因素有关。由于奶牛双胎出现了异性孪生母犊不育、胎衣不下率升高、胎间距延长、流产率和难产率升高、胎儿成活率降低等负面影响。因此，奶牛双胎对奶业生产是不利的。如果能结合性别控制等先进技术，奶牛双胎将有很大的发展前景。     

不同月龄红河黄牛血液生化和免疫指标分析

为明确放养条件下红河黄牛的营养和免疫状况，选取60头红河黄牛，公母各半，按性别和年龄（12、36和60月龄）分成6个组，颈静脉采血分离血清，测定生化和免疫指标。结果显示：①相同性别的黄牛血清总蛋白（TP）和球蛋白（GLOB）含量12月龄显著低于60月龄（P<0.05），母牛碱性磷酸酶（ALP）活性和血糖（GLU）含量12月龄显著高于60月龄（P<0.05），其他血清生化指标在不同月龄间均无显著差异（P>0.05）。公、母牛血清TP和GLOB含量均随月龄增加而升高，而ALP活性随月龄增加而降低。不同月龄母牛血清TP、白蛋白（ALB）、GLOB含量和谷丙转氨酶（ALT）活性均高于公牛（P<0.05）；而ALP活性和GLU含量公牛高于母牛（P<0.05）。②除母牛血清IL-8在12和36月龄间存在显著差异（P<0.05）外，相同性别的黄牛血清白细胞介素（IL）-2、IL-8、IL-10、免疫球蛋白（Ig） A、IgG、IgM、γ干扰素（IFN-γ）、肿瘤坏死因子α（TNF-α）含量在3个月龄间均无显著差异（P>0.05）。不同月龄母牛IL-2、IgA、IgG、IgM、IFN-γ和TNF-α含量均高于公牛（P<0.05）。综合以上结果，红河黄牛公、母牛随月龄增加，血清TP和GLOB含量显著增加，而ALP活性降低；母牛血清生化和免疫指标总体高于公牛。     

牛双胎的研究与应用

牛双胎可显著的提高生产效率，对母牛和犊牛没有明显的不良影响，不利之处是异性双胎造成母犊不孕、。双胎是可以遗传的，双亲的影响较大；不同品种间的双胎率差异显著；另外胎次、季节、营养均影响双胎率。目前常用的提高双胎率的方法是通过遗传选择、激素诱导、激素免疫、胚胎移植等方法。但如果结合标记辅助选择法（MAS）、性别控制、及转基因与克隆技术将有助于快速提高双胎率，提高生产效益。     

Herd-specific strains of Mycoplasma bovis in outbreaks of mycoplasmal mastitis and pneumonia

Mycoplasma bovis causes severe economic losses in livestock production, particularly on the Northern American continent and more recently also in continental Europe. The aim of the current study was to evaluate whether the recently emerging outbreaks were due to a particular clone or strain of M. bovis or whether these outbreaks are due to multiple infectious strains of M. bovis. The study is based on the analysis M. bovis isolated from cattle of herds with outbreaks of mycoplasmal mastitis or pneumonia from geographically non related parts of Switzerland. M. bovis isolates were typed by insertion sequence (IS) element analysis based upon ISMbov1 and ISMbov2 southern-blot hybridization. We observed a strong divergence of M. bovis strains among affected herds which mostly were herd specific. This argues against the assumption that a recent infiltration of a particular clone of M. bovis is the cause of the perilous emerging outbreaks. The study suggests that transmission occurs from animal to animal most probably via milk.     

Karyotyping, red blood cell and haemoglobin typing of the mithun (Bos frontalis), its wild ancestor and its hybrids

In Bhutan in remote parts of the Himalayas the mithun (Bos frontalis) has been used probably for centuries in an ingenious system for crossbreeding with domestic cattle which results in highly profitable hybrid females. The hybrid males are infertile and so far no stable crossbreed has been developed. Genetic analyses reported in this paper support the view that the gaur is the wild ancestor of the mithun. Both have only 58 chromosomes in contrast to 60 in cattle and also different, hitherto undescribed haemoglobins and blood groups which would justify the revision of the present classification of the subfamily Bovinae. If, with the aid of modern genetic methods, a stable crossbreed could be developed, dairy and beef production in Bhutan and many climatically similar areas could benefit greatly. Because of its remarkable size the mithun may, in the hands of enterprising breeders, also make a useful genetic contribution to beef production elsewhere.     

Prospects for increasing the utilization of cattle embryo transfer by small-scale milk and meat producers in tropical regions

Small community cattle farmers in the tropics are facing challenges to deliver quality products whilst under pressure to increase milk and beef yields per cow. These challenges could be partially met by crossbreeding Bos taurus with Bos indicus (F1) cattle utilizing embryo transfer (ET) technology. The Bos taurus infusion would increase milk production, whilst the Bos indicus influence can improve resistance to the harsh environment of the tropics. Here, individuals from existing herds can be used to produce F1 embryos which benefit from hybrid vigour. Resultant female offspring would in turn receive an F1 embryo on reaching breeding maturity. This approach would help to provide a cost-effective, systematic approach to improve productivity in dairy and beef cattle in the tropics. However, full usage of ET, including in vitro applications, in the tropics will require improvements in procedures, resources and education.     

A cross-sectional study of bovine tuberculosis in the transhumant and agro-pastoral cattle herds in the border areas of Katakwi and Moroto districts,Uganda

A study to determine the prevalence of bovine tuberculosis in the transhumant and agro-pastoral cattle herds in the border areas of Katakwi and Moroto districts in Uganda was carried out from July 2006 to January 2007 using comparative intradermal tuberculin test containing bovine and avian PPDs. A total of 1470 animals, 612 (41.6%) males and 858 (58.4%) females, 883 (60%) young, 555 (37.8%) adult and 32 (2.2%) old animals were included. The study involved a cross-sectional multistage sampling technique with random selection of individual animals from a herd. The results revealed a 1.3% overall prevalence of bovine tuberculosis in cattle herds in the study area, with a marked variation between sub-counties. The highest recorded prevalence was 6.0% in Kapujan, while no cases were recorded in Ongogonja, Magoro and Katakwi sub-counties. Distinctly different patterns in the avian-bovine reactions were also found in different sub-counties. A multivariate logistic regression showed more positive reactions (OR = 6.3; 95%CI (1.4-26.34) in females than males. BTB prevalence did not differ significantly between cattle maintained in pastoral and agro pastoral production systems. The study demonstrated a relatively low prevalence of bovine tuberculosis in local zebu cattle reared under traditional husbandry systems in Uganda, suggesting low infectiousness of the disease under such mode of production. The risk associated with the consumption of raw milk among the pastoral communities and that, the pooling of milk together from different animals is a common practice, warrants more investigation into the zoonotic transmission of tuberculosis within these communities.     

A comparison of extracted proteins of isolates of Dermatophilus congolensis by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and Western blotting.

Antigenic diversity within a collection of 18 isolates of Dermatophilus congolensis from different Continents was examined by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and by Western blotting with sera from cattle with clinical dermatophilosis using whole cell extracts obtained by three methods and one extract of extracellular products of D. congolensis. One of the methods involving the release of a lysostaphin-solubilized protein (LSP) of whole cells of D. congolensis revealed a number of discrete and easily-identifiable bands in SDS-PAGE which were found suitable for characterizing protein patterns and was, therefore, subsequently used for a comparative analysis of the proteins of all the D. congolensis isolates. Six electropherotypes (ET) of D. congolensis were identified among the 18 isolates using the protein profiles based on the presence of four protein bands at Molecular weights (MW) 62, 28, 17.4 and 16.4 kDa. The ETs were found among isolates from different animal species and from different sources with ET1 consisting of three bovine and two equine isolates; ET2, two bovine and three ovine isolates; ET3, two bovine isolates; ET4, two bovine isolates; ET5, one bovine and one ovine isolates and ET6, two bovine isolates. Immunoblotting of the extracts of D. congolensis isolates with sera from cattle with clinical dermatophilosis infection demonstrated protein bands of MW ranging from 9 kDa to 188 kDa. Sera from chronic dermatophilosis infection demonstrated a 28 kDa protein which was immunodominant in the LSP extracts of all the 18 isolates of D. congolensis tested while sera from mild infections demonstrated mainly the 62 kDa protein in the same extracts. However, many protein bands were demonstrated in surface membrane (TSMP) and extracellular protein extracts with sera from only mildly infected animals. The protein patterns observed in all isolates of D. congolensis revealed global antigenic similarities and distinct differences among isolates which could not be associated with either geographic, climatic or host factors. Also sera from infected animals from endemic regions of dermatophilosis could not differentiate isolates of D. congolensis. This suggests the possibility that such sera must have come from animals that had been infected by a multitude of D. congolensis strains present in the herd environment and strains an animal could have come across during the 'ritual' annual cross-country migration of the cattle herds.     

鲁西黄牛体细胞克隆保种技术的研究

以体外培养的鲁西黄牛耳皮肤成纤维细胞作核供体，研究利用体细胞克隆技术保存我国地方黄牛优良品种的技术方法。通过组织块培养法建立的鲁西黄牛（1♂，4♀）成纤维细胞系，作为核移植供体细胞，利用屠宰场母牛卵巢卵母细胞经体外培养成熟后作为核受体进行核移植，试验结果表明：重构胚的融合率为62．5％（242／387），分裂率为63．6％（154／242），体外培养第7天囊胚发育率为42．9％（66／154）。体外培养第7天的囊胚的内细胞团细胞（ICM）与滋养层细胞数平均为37和47，ICM占44．2％。体外发育到7d的囊胚新鲜胚胎的移植妊娠率（新鲜胚胎）移植受体10头，60d妊娠率为20％（2／10）。     

Energy metabolism of growing broiler chickens kept in groups in relation to the environmental temperature. 1. Feed intake, heat production and energy utilization]

In 5 experiments with young broiler chickens kept in groups at environmental temperatures (ET) of 35, 30, 25, 20 and 15 degrees C feed intake, live weight development and heat production were ascertained between their 5th and 57th days of life. Feed intake and live weight were ascertained separately according to sex. With live weight being the same, feed intake increased with decreasing ET. There was no maximum feed intake up to the 57th day of life at between 20 and 25 degrees C. At the end of their 57th day of life the male and the female chickens had achieved the following live weights (in the sequence of decreasing ET 35-15 degrees C): 1342; 2014; 2829; 2946; 2374 and 1367; 1900; 2512; 2496 and 2200 g/animal resp. The highest live weight gain of the chickens was achieved at between 25 and 20 degrees C with 60-70 g/d for male and with 50-60 g/d for female animals. Heat production (HP) increased progressively with age. The highest HP was registered at 20 degrees C ET.     

Characteristics of Peri-urban Dairy Herds of Bobo-Dioulasso (Burkina Faso)

Peri-urban dairy cattle farms within 50 km of Bobo-Dioulasso were studied to assess herd type, disease incidence, management, feeding and breeding strategy. Out of 417 cattle farmers, 42% had dairy objectives and were studied. Among these peri-urban dairy farmers, 60% were settled, 36% semi-settled, and 4% transhumant. In total, they held 4558 dairy cows, of which 32% lactated during the study. The prevalence of mastitis (55%) increased (p<0.05) with herd size. Advanced strategies for supplementary feeding and breeding were most frequent in small herds (<30 cattle). None of the large herds (>60 cattle) had advanced breeding strategies. Bulls and cows were culled at younger ages in herds with better breeding strategy. Overall, this resulted in higher individual milk offtake in small herds (2.46 L/day) compared with large herds (1.25 L/day). Pure breeds were rarely used, and the presence of Bos taurus baoulé naturally selected for trypanotolerance was low. The prevalence of trypanosomosis (40%) in herds dominated by Bos indicus zebu and Méré (Zebu x Baoulé) is an argument for maintenance of biodiversity and selection of Baoulé for milk production.     

Livestock keeper perceptions of four indigenous cattle breeds in tsetse infested areas of Ethiopia

Four cattle breeds indigenous to western and south-western Ethiopia - Abigar, Gurage, Horro and Sheko - were included in a study of the perceptions of smallholder cattle keepers regarding cattle management, production levels and constraints for production. A semi-structured questionnaire was used and 60 cattle keepers from each of the four areas were interviewed. Diseases were reported as the main constraint to cattle production by a majority of livestock keepers in all areas except in the Sheko area, where over-stocking was the main constraint. Among diseases, trypanosomosis was the main livestock disease according to more than half of Gurage, Horro and Sheko keepers, whereas anthrax was most important in the Abigar area. Gurage had highest age at first calving, longest calving interval and also the lowest milk production, whereas Sheko and Abigar had the most favorable characteristics both for milk production (600–700 kg) and fertility (age at first mating 3–3.5 years and above 8 calves/cow). Cattle keepers in the Sheko area reported relatively less problems with cattle diseases compared to the other areas, especially regarding trypanosomosis. Abigar showed a different disease pattern than the other breeds and may also have advantages as regards trypanotolerance.     

Herd-level risk factors for Neospora caninum seroprevalence in dairy farms in southern Brazil

A cross-sectional study was used to test the relationship between herd seroprevalence to Neospora caninum and various potential herd-level risk factors in 60 dairy farms located in two distinct regions in southern Brazil. Thirty farms were randomly selected from within each region. A questionnaire was designed to summarize each farm's production system as it might relate to N. caninum transmission. The questionnaire contained 105 closed questions relating to general characteristics of the farms, farm facilities, management, source of food and water, herd health, environment and biosecurity, which included questions relevant to N. caninum transmission, including presence and number of dogs and other animals, purchase of animals and contact with man. Serum samples were collected from 40% of animals in each farm and N. caninum antibodies were detected by immunofluorescent antibody test (IFAT). The association between potential risk factors and the probability of an animal being seropositive was modeled using a generalized estimation equations (GEE) logistic regression model. The model accounted for multilevel correlation of data from multiple animals within herds. The mean (+/-S.D.) number of animals in the 60 herds was 64.5 (+/-45.6), ranging from 20 to 280 females. Blood samples were collected from 1549 animals. The size of the farms varied from 4 to 100 ha (mean 30.1+/-25.9 ha). At least one dog was found in 57 of the 60 dairy farms (95%). The mean number of dogs was 3.1 (+/-1.9), ranging from 0 to 10. All females were raised on pasture. For all cattle sampled, N. caninum seroprevalence was 17.8%. Overall, 93.3% of herds (56/60) had at least one seropositive animal identified. Four variables were significantly associated with N. caninum sero-response in the 57 dairy farms, which were included in the final multivariable model: the number of dogs on the farm, farm area (hectares), feeding pooled sources of colostrum and region. The odds of a cow being seropositive increased 1.13 times for each additional dog present on the farm (P=0.021). Cattle from farms that fed calves colostrum pooled from multiple cows had 1.79 times greater odds for being seropositive for N. caninum (P<0.003). The probability of being seropositive was inverse to the area of the farms, such that cattle had 0.92 times the odds to be seropositive (P=0.014) for each additional 10 ha of farmland. Finally, cattle from farms in region one had 0.71 times the odds to be seropositive than cattle from region two (P=0.035). Results of this study suggest that several risk factors may explain why dairy cattle in Brazil may become exposed to N. caninum. However, further investigation of these factors is necessary because the purpose of this study was to refine and generate hypotheses on N. caninum transmission.     

Conception rate of Holstein and Japanese Black cattle following embryo transfer in southwestern Japan

This study aimed to quantify and compare conception rates to embryo transfer (ET) of Holstein and Japanese Black cattle in southwestern Japan. A 10‐year retrospective epidemiological survey was conducted. The recipient numbers for Holstein and Japanese Black cattle was 621 and 527, respectively. Conception rates of Holstein and Japanese Black cattle during the study period were 45.4% and 42.3%, respectively. There was no significant difference between both breeds. However, a different trend of conception rate to ET in Holstein and Japanese Black cattle was observed. In Holstein cattle, conception rate in August to October was lower than in the other months and was significantly lower (p < .05) than in April. Particularly, conception rate in October of Holstein cattle was the lowest (31.0%). In Japanese Black cattle, conception rates in December (24.2%) and January (31.3%) were lower than in the other months. Conception rate of Japanese Black cattle declined as the temperature–humidity index (THI) decreased, exhibiting significantly lower levels in the ≤45 THI class than in any other THI class (p < .05). By contrast, in Holstein cattle, no relationship was observed between conception rate and THI on the day of ET. These observations suggest the importance of appropriate management that considers seasonal reactivity in each breed.     

Identification of tuberculosis in cattle slaughtered in Mexico

OBJECTIVES: To determine epidemiologic factors associated with tuberculosis (TB) in dairy cattle slaughtered in 6 important regions for milk production in Mexico. ANIMALS: 2,500 cattle. PROCEDURE: Tissue specimens with lesions typical of TB were obtained during routine inspection of carcasses at abbatoirs between July 1996 and January 1997. Infection with Mycobacterium organisms was confirmed by histologic examination and bacteriologic culture. Species identification was made by use of selective growth medium, conventional biochemical tests, and radiometric procedures. Epidemiologic information for affected cattle was obtained by personal interviews with cattle dealers and owners. RESULTS: 400 (16%) of 2,500 cattle carcasses had gross lesions typical of TB. Of the 400 infected cattle, 336 (84%) had lesions in > or = 1 lymph node. Infection was confirmed in 87% of cattle with gross lesions by histologic examination, in 77% by bacteriologic culture at a laboratory in the United States, and in 59% by bacteriologic culture at a laboratory in Mexico. Most cattle were adult females in fair to good body condition that came from large herds (> 500 cattle) and were not included in the Mexican TB control program. CONCLUSIONS AND CLINICAL RELEVANCE: Mean prevalence of lesions typical of TB in dairy cattle at 6 locations in Mexico was 16%. Mycobacterium infection was confirmed by various techniques in most lesions. Recognition of typical gross lesions at slaughter may expedite TB control procedures.     

Some Aspects of Twin-gestation in the Mare

A study has been made of 44 cases of twin pregnancy in mares. The fetuses, foals, fetal membranes and blood groups of parents and foals were examined in order to determine the incidence of chorio-vascular anastomosis, blood chimaerism and freemartinism.

All of the twins proved to be dizygotic and 33 of the 34 pregnancies were bicornual. Fusion of the chorion was seen in 14 of 22 placentas and macroscopic choriovascular bridges were observed in five of these, although blood chimaerism was diagnosed in 11 of 25 cases (44 per cent) examined. This would indicate a high incidence of microchoriovascular anastomosis. The fetal female genital organs of four heterosexual twins were found to be anatomically and histologically normal at autopsy despite the existence of blood chimaerism. Five other sexvalley mature mares born as heterosexual twins with chimaerism all had normal genital organs and estrus cycles and one of these has since become pregnant. The important interspecies differences with respect to the incidence and degree of choriovascular anastomosis and freemartinism are stressed and suggestions for a plausible explanation were advanced.

     

High level expression, purification, and in vivo activity of bovine granulocyte-colony stimulating factor produced using a baculovirus system

A bovine granulocyte-colony stimulating factor (bG-CSF) cDNA clone bearing a C-terminal poly-His-tag (bG-CSFHis) was constructed and expressed by the baculovirus expression system. The bG-CSFHis was expressed as an approximately 19kDa protein in the culture supernatants and was purified using a nickel chelate column. The purified bG-CSFHis had bioactivity in vitro in the NFS-60 bioassay. In order to evaluate activity in vivo, purified bG-CSFHis was administered to cattle as single or multiple dosages. The bG-CSFHis increased neutrophil counts in peripheral blood and modulated the phagocytic activity of the neutrophils. The data indicates that the recombinant protein had activity in vivo.