鹿流行性出血病病毒高通量液相芯片检测方法的建立

为建立可检测鹿流行性出血病病毒(EHDV)的液相芯片快速检测技术,用DNAStar软件对GenBank中EHDV的VP7基因进行序列分析,设计EHDV特异性探针并用生物素标记,与荧光编码微球偶联后与病毒VP7基因的PCR产物杂交反应,用液相芯片检测仪(Liquichip200)检测荧光信号建立了EHDV快速高通量液相芯片检测方法。检测结果显示,该法具有较好的特异性,不与其他虫媒病病毒反应;检测灵敏度为100个TCID50。建立了快速检测EHDV的液相芯片技术,为进一步搭建EHDV快速高通量检测平台奠定了基础。     

奶牛流行性淋巴管炎的诊断与治疗

流行性淋巴管炎又称假性皮疽(Pseudofarcy),是由皮疽组织胞浆菌引起马属动物的一种慢性、接触性、创伤性的传染病.首次报道见于14世纪,19世纪中期已广泛流行于地中海地区,但在非洲流行最严重,故有非洲鼻疽之称.     

Association between milk production, somatic cell count and bacterial dermatoses in three dairy cattle herds

OBJECTIVE: To determine the correlations between three bacterial dermatoses in cattle, milk production and bulk-milk somatic cell count (BMSCC). DESIGN: Field observations in three dairy cattle herds. METHODS: Milk production, BMSCC, fertility and all herd diseases were recorded by computerised dairy management systems. Each herd was visited twice weekly and the clinical signs, course of diseases and morbidity and culling rates were noted. Bulk-tank milk was sampled twice monthly and analysed for somatic cell count. Bacteriological and histological examinations were carried out from samples collected from affected animals in the respective herds. RESULTS: The acute exudative form of dermatophilosis was diagnosed only in first-calving cows. The morbidity rate was 53% and the culling rate was 16% of the affected animals. The BMSCC increased by a factor of 2.4 times, and there was an average loss of milk production of 30%/cow/day in affected animals. Ulcerative lymphangitis was diagnosed in first-calving cows (22%) and older cows (15%). The culling rate was 28%. The BMSCC increased by a factor of 17.3 times, and the average loss of milk production was 5.5%/affected animal/day. Papillomatous digital dermatitis (PDD) was diagnosed in first-calving cows (25%) and in older cows (18%). The culling rate was 8%. The BMSCC increased by a factor of two times, and the average loss of milk production was 1.7%/affected animal/day. CONCLUSIONS: The correlations between three skin diseases (ulcerative lymphangitis, dermatophilosis, papillomatous digital dermatitis), milk production and BMSCC have been found to be unfavourable.     

Metazoan parasites on gills of Southern Bluefin Tuna (Thunnus maccoyii) do not rapidly proliferate after transfer to sea cages

A relatively new and highly valuable aquaculture industry focuses on three species of bluefin tunas, which are captured from the wild and fattened for several months in sea cages. In teleost aquaculture, mortalities and extra production costs are very commonly associated with metazoan ectoparasites. In tuna, however, the production value lost due to diseases associated with ectoparasites is unknown. We collected epidemiological data on burdens of metazoans on the gills of farmed southern bluefin, Thunnus maccoyii, in a series of monthly samples of tuna from the time of stocking through to harvest (March to August, 2004; N = 210) in five sea cages on a farm off Port Lincoln, Australia. Three species were recorded; for one (a copepod, Pseudocycnus appendiculatus), there was a gradual, significant increase in both abundance (from a mean of 0.1 in March, to 3.83 in August) and prevalence (from 10% to 67.5%). For the other two species (a second copepod, Euryphorus brachypterus, and a polyopisthocotylean flatworm, Hexostoma thynni) there were no discernible trends in prevalences and abundances. These results contrast markedly with those of other intensively cultured species of finfishes, in which parasite epizootics are frequent. This finding may indicate that despite the stresses of captivity, tuna mount a robust immune response to ectoparasites; the relatively low stocking densities at which tuna are farmed may facilitate this. The fall in water temperature during farming (22 °C to 13 °C) may also reduce the reproductive rate of these ectoparasites.     

Identification of the etiologic agent of epizootic bovine abortion in field-collected Ornithodoros coriaceus Koch ticks

Epizootic bovine abortion (EBA), or foothill abortion as it has often been termed, is a tick-borne disease of pregnant cattle recognized in California, Nevada and Oregon. The primary objective of this study was to better define the relationship of a novel deltaproteobacterium, the putative etiological agent of EBA (aoEBA), with the Pajaroello tick (Ornithodoros coriaceus Koch), the recognized vector of EBA. Three developmental stages of O. coriaceus (larva, nymph, and adult) were collected from five locations in California, Nevada and Oregon. A polymerase chain reaction (PCR), developed for detection of aoEBA, was applied to DNA extracted from ticks. Southern blotting of the PCR products increased the number of ticks determined to be carrying the bacteria by seven-fold, suggesting the majority of infected ticks carry relatively low numbers of the pathogen. An effort was made to determine if an artificial blood meal would stimulate replication of the bacterial pathogen, thereby increasing the frequency in which aoEBA could be identified; no statistically significant effect was evident. The number of ticks determined to be carrying aoEBA varied with geographic location and ranged from 5 to 20%. aoEBA was found in both adults (12% of the males and 12% of the females) and nymphs (13%) but not larvae. Comparative analysis of dissected ticks provided strong evidence that the salivary gland was the most common location of aoEBA in field-collected ticks. No significant correlations were identified between the frequency of infection and tick weight, suggesting that increasing tick age and increased number of blood meals did not increase infectivity.     

Infectious agents associated with epizootic rabbit enteropathy: isolation and attempts to reproduce the syndrome

Epizootic rabbit enteropathy (ERE), a highly lethal (30-80% mortality) disease of broiler rabbits aged 6-14 weeks, first appeared in 1997 in French intensive enclosed rabbitries and is of unknown aetiology. Bacteriological, virological and parasitical examination of the intestinal contents of rabbits that had died either in spontaneous field cases or after experimental reproduction of ERE, were undertaken in an attempt to identify infectious agents that may play a role in the disease. Two bacterial strains, Clostridium perfringens and non-enteropathogenic Escherichia coli were repeatedly isolated at high faecal counts from naturally infected animals. In field cases, a correlation between typical gross lesions of epizootic enteropathy and the presence of the alpha toxin of Cl. perfringens was observed (P<0.0001; Chi-squared test). Although attempts to reproduce the disease by inoculation with different pools of cultivable bacterial strains failed, the disease was successfully reproduced by inoculation with one French and two Belgian samples of caecal contents.     

鹿流行性出血病病毒VP7基因的克隆及原核表达

根据已经扩增的鹿流行性出血病病毒(EHDV)VP7基因序列,设计一对扩增VP7基因特异性引物,用RT-PCR从血清I型EHDV(EHDV-1)总RNA中扩增VP7基因,并将其克隆到原核表达载体pBAD/Thio中,构建了重组原核表达质粒pBAD/Thio-EHDV VP7。将表达载体转入大肠埃希菌(E.coliLMG194),用L-阿拉伯糖诱导表达。SDS-PAGE和Western blot试验结果表明,EHDV VP7基因在LMG194中得到了表达,其表达产物可以与EHDV阳性血清特异性反应,说明该融和蛋白具有免疫学活性。     

鹿流行性出血病病毒双抗夹心ELISA方法的建立

为建立鹿流行性出血病病毒(EHDV)病原学检测方法,用纯化的抗EHDV特异性单克隆抗体包被ELISA板,用兔抗EHDV IgG作为夹心抗体,IgG作为夹心抗体建立EHDV双抗夹心ELISA方法,并对该方法的特异性和敏感性进行了试验.用ELISA分别检测EHDV、蓝舌病病毒(BTV)、水疱性口炎病毒(VSV)、赤羽病病毒...