单克隆抗体ELISA检测犊牛粪便中冠状病毒抗原

用两株单克隆抗体混合后包被ＥＬＬＩＳＡ微孔板，加牛冠状病毒抗原，加经白陶土处理过的兔抗牛冠状病毒免血清，再加上辣根氧化物酶标记的羊抗兔ＩｇＧ抗体，加底物质显色间接ＥＬＩＳＡ检测牛冠状病毒抗原获得成功。用此方法检测细胞培养的牛冠状病毒上清液，其敏感度高出血凝试验（ＨＡ）８倍，从武汉市附近３个奶牛场采集犊牛腹泻粪便１０５份，用单抗ＥＬＩＳＡ检测牛冠状病毒抗原，共检出阳性３６份，并与血凝及血凝抑制试验，     

Studies on efficacy and stability of a vaccine bait containing ERA strain of rabies virus propagated in a BHK-21 cell line.

In a dose response study in foxes, the median protective dose of ERA BHK21 vaccine in a blister pack bait was 10(6.0) tissue culture infective doses (TCID)/mL, while artificially aged baits with titers of 10(6.3) TCID/mL induced seroconversion in 78% of foxes. There was no significant difference in the development of antibodies in foxes receiving 1, 2 or 3 mL volumes of vaccine in the bait. When baits were exposed to the elements and fed to foxes over a 21 day period, 85% of the animals seroconverted. Age, sex and the way in which the vaccine container was contacted did not appear to be factors in the responses of these animals. Juvenile foxes, approximately six months of age, were marked more readily with the tetracycline bait marker than older animals. Approximately 25% of foxes did not appear to respond well to vaccination and the titer of the vaccine was a critical factor in producing seroconversion in these animals.     

Behavioral and physiological effects of freeze or hot-iron branding on crossbred cattle.

Twenty-seven crossbred calves (1/2 Simmental, 1/4 Hereford, 1/4 Brahman) averaging 257 +/- 11 d of age were either hot-iron-branded (H), freeze-branded (F), or sham-branded (S). Calves were blocked for temperament, weight, and sex and were randomly assigned to day and order in which treatments were applied. To reduce stress from handling at treatment time, each calf was herded through the squeeze chute daily for 5 d before the experiment. Jugular cannulas were inserted in each calf 1 d before application of treatment. Blood samples and heart rate measures were obtained at -5, -3, 0, .5, 1, 3, 5, 10, 15, and 20 min after application of the treatments. Mean concentrations of plasma epinephrine (EPI) were higher for H calves at time .5 min than for either S or F calves (P = .10). To account for individual differences, prebranding heart rates and hormone concentrations were subtracted from subsequent samples and were also used to calculate a proportion for each subsequent sample. Analyses of subtracted values found that EPI concentrations were greater for H calves than for either S or F calves (P = .007) at .5 min postbranding. No other differences were found for the subtracted analyses. Analyses of proportion data also revealed that H calves had greater EPI than did either S or F calves (P = .027) at .5 min postbranding. Only three animals vocalized during branding, one H calf and two F calves. Despite the 5-d acclimation period, handling and restraint elevated plasma cortisol concentrations and heart rate.(ABSTRACT TRUNCATED AT 250 WORDS)     

Occurrence of Staphylococcus intermedius on the hair and skin of normal dogs.

Aerobic bacterial populations were studied on the distal hair coat and at the skin surface of the shoulder, rump and abdomen of 10 healthy dogs. Coagulase negative staphylococci (CNS) were more frequently isolated from the hair than the skin at the shoulder and rump. There was no difference in the isolation rate of coagulase positive staphylococci (CPS) (Staphylococcus intermedius) between the hair and skin. Total skin counts were greatest on the abdomen whereas CNS counts from the hair were least at this site. There were no differences between CPS counts at the three sites on either hair or skin. The populations on the relatively unfavourable microenvironment of the distal hair may represent contamination rather than colonisation. The low populations of CPS at the skin surface also indicate contamination or transient colonisation rather than true resident status.     

Studies on mutlispecific resistance of gastrointestinal nematodes to benzimidazoles on dairy-goat farms

Multispecific resistance to benzimidazoles was studied in three selected farms. These farms had bred dairy goats for more than 15 years. The helminths were introduced with the goats at the establishment of the farms which afterwards remained isolated. Nematode resistance could then be related to their own management practices. Faecal egg count tests and egg hatch assays were performed to assess intensity of resistance. The generic (infective larvae in faecal cultures) and specific richness (adult worms) were assessed. The resistant species were Trichostrongylus colubriformis, Teladorsagia circumcincta, Haemonchus contortus and Oesophagostomum venulosum. Faecal egg count reduction tests and egg-hatch assays did not match exactly. Faecal larval counts after treatments gave a distorted picture of multispecific resistance: Haemonchus and Oesophagostomum were very largely over represented. The number of species found in the three farms was relatively low compared with other reports in goat farms of the area. This reduction of diversity might also be due in part to characteristics of breeding management and history (use of permanent pasture and introduction of goats at the establishment of farm).     

Prevalence of mycoplasmas in the respiratory tracts of pneumonic calves.

The prevalence of mycoplasmas in the respiratory tracts of 148 pneumonic calves originating from 25 herds in the Netherlands is reported. Four types of culture media were used to isolate mycoplasmas: solid modified Edward medium, 2 types of Friis media, and A7B differential agar medium. Mycoplasmas were isolated both from nasal swab specimens and lung lavage fluids collected from live calves and from nasal mucosa and lung tissue specimens collected post mortem. All of the mycoplasma strains isolated could be identified as either Ureaplasma diversum (isolated from 80% of 25 herds), Mycoplasma dispar (92%), M. bovirhinis (88%), M. bovis (20%), M. bovigenitalium (4%), M. arginini (16%), or M. canis (12%). Isolation rates of M. dispar and U. diversum were considerably higher from lung lavage fluids than from nasal swab specimens. M. bovis was detected only in fattening herds and not in dairy herds. The respiratory tracts of 75% of the calves examined contained at least 2 mycoplasma species. In total, 25 different combinations of mycoplasma species were detected in specimens collected from noses and lungs. The pathogenic species U. diversum and M. dispar had not been isolated before in the Netherlands.     

Genomic variability among globally distributed isolates of equine arteritis virus.

Equine arteritis virus (EAV), a non-arthropod borne togavirus, has been shown to have a global distribution. To date, no major antigenic variation has been demonstrated between EAV isolates from different geographic origins. In this study, the genomic RNA of EAV isolates obtained from horses of different breeds in various countries around the world was oligonucleotide fingerprinted. Comparisons of these fingerprints were used to determine the extent of genomic variation among such isolates. Comparisons among isolates from North American horses revealed, for the most part, oligonucleotide homologies of less than 60%. Only 29 of the 98 comparisons revealed greater than 60% oligonucleotide homology. Nonetheless, several comparisons indicated a close epidemiologic relationship between isolates from horses of different breeds located in different states. Though all European isolates were of Standardbred origin and were from horses located in northern European countries, the majority had oligonucleotide homologies of less than 60%. Where oligonucleotide homology was apparent, it was, with one exception, greater than 70%. The two isolates from New Zealand had 93.2% oligonucleotide homology. This is indicative of an extremely close epidemiologic relationship. Comparisons between EAV isolates from around the world revealed oligonucleotide homologies between viruses from North America, Europe and New Zealand. In several instances, this homology was greater than 70% and in one case greater than 80%. No oligonucleotide homology was evident in comparisons involving the virus from South Africa. The high level of genomic conservation between certain EAV isolates of disparate geographic origins may reflect dissemination of the virus associated with the international movement of horses. The extent of genomic variation demonstrated between most of the EAV isolates used in this study confirms the need for further investigation of genomic heterogeneity among strains of this virus before techniques that rely upon nucleic acid hybridization can be effectively applied as diagnostic procedures.     

Verification by polymerase chain reaction of vertical transmission of Theileria sergenti in cows

To evaluate the transplacental transfer of Theileria sergenti infection in cattle, we used DNA probes to detect T. sergenti in 6 pregnant cows and their calves. All the animals were monitored by parasitologic, serologic, and polymerase chain reaction (PCR) assays for a predicted 875-base-pair (bp) DNA product and a 684-bp amplicon detected by nested PCR in the blood and spleens of aborted fetuses. An open reading frame (ORF) starting at nucleotide 170 and terminating at position 1021 was shown to code for a polypeptide of 283 amino acid residues. All 6 dams and 5 calves were positive for T. sergenti in all tests. One calf was positive only with nested PCR. We conclude that transplacental transmission of T. sergenti is a significant problem. The relevance of the data in the programmed introduction of new (especially pregnant) animals into established clean herds needs serious consideration with regard to control of theileriosis and other tickborne diseases.     

畜禽注水肉的卫生检验与卫生处理

注水肉系指不法经营者无视国家的屠宰法规和食品卫生法规 ,对畜 (牛、猪、兔等 )、禽 (鸡、鸭 )等在宰前或宰后人为地采用某种手段器械(注射器、皮管、压力泵 )通过畜禽口腔、食道、肛门直肠、动脉血管、皮下等部位注射入一定量的水分 ,以此增加畜禽重量 ,使胴体、肌肉、内脏的含水量剧增 ,达到饱和状态。近几年来 ,畜禽注水肉多见于牛肉、猪肉、鸡鸭肉 ,尤其在农贸集市非常普遍 ,已引起广大消费者和有关部门的关注。１畜禽注水的途径与方法１．１宰前活体注水灌食强行固定猪体灌水姿势 ,用皮管塞入口腔或肛门注入水 ,注水量可占体重的２０%…     

Manipulated Mouse Embryos as Bioassay System for Water Quality Control

Mouse pronuclear stage embryos with intact slit zona pellucida (manipulated) were cultured in vitro until the hatched blastocyst stage in simplex optimized medium with higher K⁺ concentration (KSOM) prepared with three different water types: tap, deionized reverse osmosis (D‐O) water and Milli‐Q system (M‐Q) water. The culture media were supplemented with or without protein and ethylenediaminetetraacetic acid (EDTA, disodium salt). The rates of hatched blastocysts were significantly affected (p < 0.01) by micromanipulation, protein supplement and water source. The water source has no influence (p > 0.05) on development in EDTA‐supplemented protein‐free culture media, whereas in EDTA‐ and protein‐free culture media, the water quality significantly (p < 0.001) affected the rates of development, with higher rates in media prepared with M‐Q water. The micromanipulated embryos showed higher sensitivity to the water quality (p < 0.01). It worth mentioning that the rates of hatched blastocysts in protein‐free culture media were very low (0–7.5%). Furthermore, the three different water types were analysed by measuring the electrical conductivity, inorganic ions, total organic carbon and endotoxins to evaluate the purity. M‐Q water showed the lowest levels of inorganic ion, total organic carbon and endotoxin concentrations. We concluded that manipulated mouse embryos are good system to evaluate the quality of water used in biological system.