Detection of foot-and-mouth disease virus infected cattle using infrared thermography

In this study, infrared thermography (IRT) was assessed as a means of detecting foot-and-mouth disease virus (FMDV)-infected cattle before and after the development of clinical signs. Preliminary IRT imaging demonstrated that foot temperatures increased in FMDV-infected animals. The maximum foot temperatures of healthy (n = 53), directly inoculated (DI) (n = 12), contact (CT) (n = 6), and vaccine trial (VT) (n = 21) cattle were measured over the course of FMD infection. A cut-off value was established at 34.4 °C (sensitivity = 61.1%, specificity = 87.7%) with the aim of detecting FMDV-infected animals both before and after clinical signs were observed. Seven of 12 (58%) DI and 3/6 (50%) CT animals showed maximum foot temperatures exceeding the 34.4 °C cut-off before the development of foot vesicles. In contrast, only 5/21 (24%) VT animals displayed pre-clinical foot temperatures above this cut-off possibly indicating partial vaccine protection of this group. These results show IRT as a promising screening technology to quickly identify potentially infected animals for confirmatory diagnostic testing during FMD outbreaks. Further evaluation of this technology is needed to determine the value of IRT in detecting animals with mild clinical signs or sub-clinical infections.     

Simulation study to assess the efficiency of a test-and-cull scheme to control the spread of the bovine viral-diarrhoea virus in a dairy herd

To control the spread of bovine viral-diarrhoea virus (BVDV), test-and-cull schemes have been used in Scandinavian countries, with success, when combined with strict control of new animal introductions into herds. In situations where BVDV reintroduction is likely to occur, it is necessary to assess precisely the expected efficiency of test-and-cull schemes. The objective of this study was to compare, by simulation, the persistence and consequences of BVDV infection in a fully susceptible dairy herd with either a test-and-cull scheme or no control action. We used a stochastic individual-based model representing the herd structure as groups of animals, herd dynamics, the contact structure within the herd and virus transmission. After an initial introduction of the virus into a fully susceptible herd, the frequency of purchases of animals that introduced the virus was simulated as high, intermediate or null. Virus persistence and epidemic size (total number of animals infected) were simulated over 10 years. The test-and-cull reduced the epidemic size and the number of days the virus was present except in herds with complete prevention of contact between groups of animals. Where no virus was reintroduced, virus persistence did not exceed 6 years with a test-and-cull scheme, whereas the virus was still present 10 years after the virus introduction in some replications with no control action (<2%). Where frequent purchases were made that led to virus introduction (6 within 10 years), with an intermediate virus transmission between groups, the probability of virus persistence 10 years after the first virus introduction fell from 31% to 8% with the test-and-cull scheme (compared to the do-nothing strategy). Within the newly infected herd, the test-and-cull scheme had no effect, on inspection, on the number of PI births, embryonic deaths or abortions over 10 years. Given this, the economic efficiency of the test-and-cull scheme should be further investigated.     

Bold fusion: transtracheal wash from a Jersey calf

A 10-week-old, female, Jersey calf was referred to the University of Minnesota Veterinary Medical Center for evaluation of difficult breathing and inappetence of 18-hours duration. Cytologic examination of a transtracheal wash specimen revealed pyogranulomatous inflammation, with increased numbers of multinucleated cells. Differential diagnoses included established bacterial infection, viral infection, fungal infection, and foreign body reaction. Immunocytochemical staining was done on a direct smear of the transtracheal wash using a cocktail of 4 mouse monoclonal antibodies against 3 human respiratory syncytial virus proteins. Strong intracytoplasmic staining within mononuclear cells was observed and a diagnosis of bovine respiratory syncytial virus infection was made. Immunocytochemistry may be used as a rapid, inexpensive, adjunctive diagnostic tool for the antemortem diagnosis of bovine respiratory syncytial virus on transtracheal wash specimens.     

Neospora caninum antibodies in commercial fetal bovine serum

The protozoan parasite Neospora caninum is a major cause of abortion in cattle throughout the world. In the process of propagating Neospora in vitro and producing specific antibodies for development of diagnostic assays in the food supply, our laboratory identified the presence of bovine antibodies to N. caninum in fetal bovine sera. The sera were produced commercially and preferentially recommended for tissue culture use and monoclonal antibody production. Seventeen different fetal bovine serum samples of different grades and from four different companies were examined for the presence of total IgG, IgG₁, IgG₂, and IgM specific for N. caninum. All of the tested serum samples recognized N. caninum specific bands on Western blot. Low IgG serum also recognized these antigens but with lower intensity. Antibody response was also evaluated using a commercially available ELISA kit for N. caninum.     

不同化学激活方法及卵丘细胞层数对牛体外成熟卵母细胞孤雌发育的影响

为探索适宜的牛体外成熟卵母细胞孤雌激活方式,提高核移植效率及研究胚胎孤雌发育。研究比较了不同浓度的乙醇( EH)、离子霉素(ionomycin)、钙离子载体A23187与蛋白激酶抑制剂6-二甲氨基嘌呤(6-DMAP)、细胞松驰素(cytochalasin B, CCB)对牛卵母细胞激活发育的影响,并在相同条件下,比较了不同卵丘细胞层数对卵母细胞激活后胚胎发育能力的影响。结果表明: (1) 5 μmol·L-1 Ionomycin,10 μmol·L-1 A23187,7% EH分别联合2 mmol·L-1 6-DMAP,CCB均可以有效地激活牛孤雌胚,其中以Ionomycin＋6-DMAP+CCB组囊胚发育率极显著高于其他各组(P<0.01),并且激活液中CCB的存在对牛孤雌胚胎的发育有利;(2) 卵母细胞包被的卵丘细胞层数不同对卵母细胞成熟激活有显著的影响,卵丘细胞层数3～5层和多于6层的卵丘—卵母细胞复合体(COCs)孤雌激活的分裂率和囊胚率分别为69.09%,31.58%和75.14%,38.85%,这2组之间差异不显著(P>0.05), 但其显著高于1～2层组与混合组(P< 0.05), 因此,这2组细胞是最佳的试验研究材料。     

牛脂联素cDNA全基因序列的克隆

[目的]体外克隆牛脂联素基因的编码全序列。[方法]以成牛尾部脂肪的总RNA为模板,通过RT-PCR扩增获得牛脂联素基因cDNA的全序列,将其克隆到原核表达载体PMD18-T中,筛选出阳性克隆,提取重组质粒,通过限制性内切酶酶切鉴定和重组质粒PCR鉴定后对其进行测序。[结果]从牛尾部脂肪组织总RNA中扩增得到671bp的目的条带,该cDNA序列与GenBank中牛脂联素基因序列同源性为100％,其氨基酸同源性也达到100％,确定为目的基因。[结论]该研究为进一步研究牛脂联素基因的功能和研究牛脂联素基因的表达与育肥牛的脂肪代谢的关系提供了试验基础。     

奶牛卵母细胞体外受精和胚胎培养研究

[目的]探讨适合于奶牛体外胚胎生产的最佳条件。[方法]以奶牛卵母细胞为试验材料,研究不同温度、运送时间对奶牛卵母细胞成熟的影响,并比较在相同培养条件下孤雌激活和体外受精对奶牛卵母细胞胚胎发育的影响。[结果]卵巢在37—42℃运输后的成熟率（11．3％）明显低于20—30℃、30-37℃的成熟率（79．3％,85．1％）,20～30℃、30～37℃组间差异不显著;运送时间6h后的卵母细胞成熟率（53．2％）明显低于4h内、4—6h的成熟率（85．6％,77．8％）,4h内、4～6h差异不显著;孤雌激活和体外受精胚胎在相同条件下的卵裂率（72．41％,68．18％）、8细胞率（45．24％,47．22％）、囊胚率（25．60％,18．89％）差异均不显著。[结论]卵巢在30℃左右4h内运回实验室最适宜,而孤雌激活和体外受精胚胎在相同条件下发育率差异不显著。     

PDE3与亮甲酚蓝在绵羊卵母细胞体外培养中的应用研究

[目的]探讨磷酸二酯酶3（PDE3）特异性抑制剂Milrinone与亮甲酚蓝染色液（BCB）相结合在绵羊卵母细胞体外培养中的应用。[方法]比较了BCB对绵羊卵母细胞的筛选与传统的形态分级之间的差异,以及对胚胎后期发育的影响;研究了Milrinone对绵羊卵母细胞的最佳抑制时间,并用于BCB-卵母细胞体外培养。[结果]A、B级卵母细胞中BCB＋卵母细胞的比例为64.42%,极显著高于C级卵母细胞的17.00%;BCB＋卵母细胞的成熟率（86.16%）、卵裂率（85.29%）、囊胚率（34.40%）分别极显著高于BCB-卵母细胞（50.94%、36.19%和6.73%）;6 h是Milrinone对绵羊卵母细胞的最佳抑制时间,该试验组体外培养效率显著高于其他组;Milrinone对BCB-卵母细胞抑制培养6 h,极显著提高了体外胚胎发育率。[结论]为提高绵羊卵母细胞体外培养效率提供了依据。     

A morphological assessment system for ‘show quality’ bovine livestock based on image analysis

Morphological assessment is one important parameter considered in conservation and improvement programs for bovine livestock. This assessment process consists of scoring an animal based on its morphology and is normally carried out by highly qualified staff. These animals are all of agreed ‘show quality’ and hence they are morphologically very similar.This paper presents a system designed to provide an assessment based on a lateral image of the cow. The system consists of two main parts: a feature extraction stage, to reduce the information on the cow in the image to a set of parameters, and a neural network stage to provide a score based on that set of parameters. For the image analysis section, a model of the animal is constructed by means of the point distribution model (PDM) technique. Later, that model is used in the searching process within each image, which is implemented using genetic algorithms (GAs). As a result of this stage, the vector of weights that describe the deviation of the given shape from the mean is obtained. This vector is used in the second stage, where a multilayer perceptron is trained to provide the desired assessment, using the scores given by experts for selected cows.The system has been tested with 138 images corresponding to 44 individuals of a special rustic breed, with very promising results, given that the information contained in only one view of the cow can not be considered complete.     

聚蔗糖水凝胶的制备及其在药物控制释放中的应用

[目的]研究聚蔗糖水凝胶的制备工艺,并考察制备工艺对溶胀度和牛血清蛋白（BSA）释放性能的影响。[方法]以环氧氯丙烷为交联剂,在碱性条件下制备聚蔗糖水凝胶,用BSA作为模型药物考察其释放性能。[结果]当聚蔗糖溶液浓度为16.7 g/L,交联剂用量为0.85 mol/L,反应温度为45℃,反应时间为5 h时,制得的聚蔗糖水凝胶弹性和硬度较好,其溶胀度在5.5～7.5。聚蔗糖水凝胶对BSA的释放性能表明,聚蔗糖水凝胶的交联程度决定BSA的释放速率,适当增加聚蔗糖水凝胶的交联程度有利于药物的缓慢释放。[结论]可以通过调控聚蔗糖水凝胶的溶胀度调节包埋在此水凝胶中BSA的释放速率。