猪水泡病病毒结构蛋白基因的克隆及其蛋白二级结构和淋巴细胞表位的预测

以免疫信息学和反向疫苗学为理论根据，利用RT-PCR法获得了猪水泡病病毒（SVDV）HK／70株的基因组序列并测序，应用生物信息学相关软件和方法，对SVDV结构蛋白的二级结构的不同方面及其抗原特异性淋巴细胞表位进行了预测，综合评价了SVDV结构蛋白的抗原特异性细胞表位，并计算出一些潜在的抗原位点。结果表明，VP1蛋白含有的细胞优势抗原表位最多，但其他结构蛋白也含有抗原特异性淋巴细胞表位，有的甚至有可能成为优势抗原表位或对优势抗原表位有协同作用。     

猪的应激与防控

研究猪不良的应激作用机制以及所引发的各种疾病,通过临床疾病综合症的分析,总结出紧急应激处理、加强营养与管理和培育抗应激品种等一套有效的防控措施以指导生产。     

Epidemiological studies of pig diseases: 1. Use of French protocols for risk factor assessment to predict the health status of Australian herds

Objective To determine in Australian pig herds the accuracy of French protocols for risk factor assessment.
Procedure Data on health indicators and risk factors were collected for three syndromes, 'pre-weaning diarrhoea', 'post-weaning diarrhoea' and 'respiratory problems', using the French protocols. The protocols were used on 118 occasions in 32 Western Australian pig herds during 3 years (1988 to 1991).
Results There was a wide variation in pre-weaning performance, for example growth rate was 107 to 273 g/day (< 200 g/day in 33% of herds). Respiratory lesions at weaning were associated with poor pre-weaning performance. Post-weaning (21 days after weaning) growth rate was 114 to 408 g/day (< 250 g/day in 54% of herds). In the grower herds, 91% of herds had pneumonia, and growth rate was 439 to 625 g/day (< 550 g/day in 54% of herds). Pleurisy as well as pneumonia was associated with reduced growth rate. The risk factor most closely associated with respiratory health status was air volume per pig.
Conclusion Risk factors were most accurate at predicting the health status in post-weaning problems. A weaning weight of at least 7.9 kg and weaning age of 30 days optimised weaner performance. Stocking densities and shed designs providing at least 3 m³ air volume and 0.6 m² floor space per pig throughout the growing phase should be considered for an improved respiratory health status. Australian pig sheds often do not provide a satisfactory environment for optimum health. The technique of risk factor assessment as an aid to the maintenance of health in pig herds is applicable in Australia, but further research is necessary to determine the most important Australian risk factors.     

基于UPLC-MS/MS同时测定猪粪便中5种兽药残留量的研究

为了建立同时测定猪粪便中二甲氧苄啶、磺胺对甲氧嘧啶、恩诺沙星、金霉素、泰乐菌素5种兽药残留量的超高效液相色谱-串联质谱(UPLC-MS/MS)分析方法,样品经McIllvaineNa_2EDTA缓冲溶液提取,C18粉吸附杂质,HLB固相萃取小柱净化,浓缩富集后用0.1%甲酸甲醇溶液复溶,过滤上机,基质液外标法定量。结果显示,5种兽药在10~500μg/kg的质量浓度范围内线性良好,在添加浓度50~500μg/kg,平均回收率为53%~85%,RSD为1.87%~18.81%。该方法具有较好的准确度与精密度,能快速、准确测定猪粪便中5种兽药残留,为养殖环节实时监测兽药使用情况提供了技术支持。     

高效液相色谱-串联质谱法测定猪毛中盐酸可乐定和盐酸赛庚啶

建立了高效液相色谱-串联质谱法测定猪毛中盐酸可乐定和盐酸赛庚啶的方法。猪毛样品用0.1 mol/L NaOH溶解后,用0.1 mol/L HCl溶液调节pH值至7.0,经乙腈提取后用UPLCMS/MS进行分析。分析时采用Waters BEH C18色谱柱(100 mm×2.1 mm,1.7μm),以0.10%甲酸水/甲醇作为流动相进行梯度洗脱,电喷雾正电子(ESI~+)模式电离,多反应监测(MRM)模式检测,外标法进行定量。结果表明:盐酸可乐定和盐酸赛庚啶标准溶液在1.0~50μg/L范围内,峰面积与含量呈线性相关。盐酸可乐定和盐酸赛庚啶的检出限均为0.20μg/kg,样品中添标平均回收率在85%~110%之间,相对标准偏差(RSD)小于10%。     

9种中药成分对猪传染性胃肠炎病毒体外增殖的抑制作用

运用MTT法结合细胞病变方法测定黄芩苷、穿心莲内酯、大黄素、大黄酸、芦荟大黄素、大黄素甲醚、苦参碱、氧化苦参碱和苦马豆素9种中药成分对猪传染性胃肠炎病毒(TGEV)在ST细胞上增殖的抑制作用,实时荧光定量PCR(Real-time PCR)进一步检测黄芩苷对TGEV在ST细胞上增殖的抑制作用。结果显示,9种中药成分在体外对TGEV在ST细胞上的增殖均有一定的抑制作用,其中黄芩苷对TGEV的生物合成、吸附保护和直接灭活作用的抑制率分别为58.23%、88.12%和100.00%;Real-time PCR检测黄芩苷3种给药方式均能在转录水平显著降低TGEV mRNA的相对表达量,与病毒对照组相比差异极显著(P0.01)。结果表明,9种中药成分在体外对TGEV在ST细胞上的增殖均有一定的抑制作用,以黄芩苷作用最好。     

LC-MS/MS measurement of ampicillin residue in swine tissues at 5 days after in-feed administration

We assessed ampicillin (ABPC) concentrations of kidney, muscle and intestine after a 5–day withdrawal period in 2 male and a female young Large White pigs fed the diet containing ABPC (ABPC medicated feed, 24 mg/kg/day) for a week. The ABPC residues were measured with liquid chromatography–tandem mass spectrometry, and the mean recoveries and quantitation limits ranged from 91.8 to 97.2% and from 0.1 to 0.12 ng/g, respectively. The residual ABPC concentrations were ≤1.18 ng/g for the muscle, ≤0.53 ng/g for the kidney and ≤1.93 ng/g for the intestine, suggesting below the Japanese provisional maximum residue limits. These results reveal that the analytical method is developed for residual ABPC and that the withdrawal period is appropriate.     

Porcine circovirus type 2 decreases the infection and replication of attenuated classical swine fever virus in porcine alveolar macrophages

Recently, it has been noted that porcine circovirus type 2 (PCV2) infection adversely affects the protective efficacy of Lapinized Philippines Coronel (LPC) vaccine, an attenuated strain of classical swine fever virus (CSFV), in pigs. In order to investigate the possible mechanisms of the PCV2-derived interference, an in vitro model was established to study the interaction of LPC virus (LPCV) and PCV2 in porcine alveolar macrophages (AMs). The results showed that PCV2 reduced the LPCV infection in AMs and the levels of PCV2-derived interference were dose-dependent. The PCV2-derived interference also reduced the replication level of LPCV in AMs. The full-length PCV2 DNA and its fragment DNA C9 CpG-ODN were involved in the reduction of LPCV infection in AMs, whereas UV-inactivated PCV2 was not. In addition, a moderate negative correlation between the LPCV antigen-containing rate and IFN-γ production was observed, and had a dose-dependent trend with the level of PCV2-inoculation. The results of the present study may partially explain how PCV2 infection interferes with the efficacy of LPC vaccine.     

Drivers and risk factors for circulating African swine fever virus in Uganda, 2012–2013

We explored observed risk factors and drivers of infection possibly associated with African swine fever (ASF) epidemiology in Uganda. Representative sub-populations of pig farms and statistics were used in a case-control model. Indiscriminate disposal of pig viscera and waste materials after slaughter, including on open refuse dumps, farm-gate buyers collecting pigs and pig products from within a farm, and retention of survivor pigs were plausible risk factors. Wire mesh-protected windows in pig houses were found to be protective against ASF infection. Sighting engorged ticks on pigs, the presence of a lock for each pig pen and/or a gate at the farm entrance were significantly associated with infection/non-infection; possible explanations were offered. Strict adherence to planned within-farm and community-based biosecurity, and avoidance of identified risk factors is recommended to reduce infection. Training for small-scale and emerging farmers should involve multidimensional and multidisciplinary approaches to reduce human-related risky behaviours driving infection.     

Up-regulated expression of PD-1 and its ligands during acute Classical Swine Fever virus infection in swine

In order to investigate the relationship between the PD-1 pathway and impairment of immune responses with the CSFV infection, the mRNA expression of PD-1 and its ligands were evaluated by quantitative polymerase chain reaction (qPCR) during artificial CSFV infection. Simultaneously, expression of IL-2 and IL-10 mRNA were detected. The T cell proliferation and CSFV load in plasma were also measured. Results showed that the expression of PD-1 and its ligands mRNA were significantly increased (p < 0.01) in PBMC from 3 to 7 days post infection (dpi). Meanwhile the level of IL-10 was up-regulated (p < 0.01). The IL-2 mRNA was not obviously changed but it is significantly increased from 14 dpi. The T cell proliferation was notably decreased at 7 dpi. The CSFV load was also increased in plasma. Overall, our results suggest that the expression of PD-1 and its ligands were up-regulated and probably correlated with immune inhibition during acute CSFV infection.