江苏地区2株山羊痘病毒的分离与鉴定

为对江苏部分地区羊场羊痘病毒进行有效的监测和防控,及时了解病毒遗传进化趋势,降低规模化养羊业的风险,本研究从江苏部分地区养羊场采集疑似羊痘病羊的皮肤丘疹、水疱或脓疱组织病料样品,分别接种9日龄SPF鸡胚绒毛尿囊膜、胎羊睾丸细胞(LT)和牛肾细胞(MDBK),观察鸡胚病变和细胞病变情况。同时通过PCR方法扩增羊痘病毒p32基因进行序列分析,并进行p32基因限制性片段长度多态性分析。结果显示,接种鸡胚绒毛尿囊膜形成典型的痘斑,胚体出血;系统进化分析显示,两株分离株属于山羊痘病毒(GTPV);限制性片段长度多态性分析结果也显示符合GTPV的特征。本研究所分离到的2株GTPV,为当地制定山羊痘的防控策略提供依据。     

山羊痘病毒的分离与鉴定

对贵州省2002年以来发生的疑似山羊痘病例样品进行了病毒的分离与鉴定。取山羊痘病羊的皮肤痘疹和水疱作待检病料,接种BHK-21传代细胞盲传3代后,出现了明显的、规律的细胞病变(CPE),病毒细胞培养物在F4代以后,能与山羊痘标准阳性血清在琼脂扩散试验中出现白色沉淀线,而与正常细胞的培养物及PBS不出现沉淀线;参照GenBank上山羊痘病毒P32基因序列,设计了1对特异性引物,对现场分离毒株进行PCR扩增,可扩增出963bp特异性的DNA条带。用待检病料感染的BHK-21细胞培养物接种9日龄鸡胚绒毛尿囊膜,随着传代次数的增加,痘斑病变的出现率从13.3%～20%上升至33.3%～40%;用山羊痘病变皮肤、鸡胚绒毛尿囊膜和感染细胞进行超薄切片,在电子显微镜下可以观察到典型的山羊痘病毒粒子。     

鸡传染性法氏囊病的实验室诊断

采集病死鸡法氏囊经处理后绒毛尿囊膜途径接种5枚SPF鸡胚,接种胚均于接种后48~96 h死亡,死亡胚病变特征为体表和皮下出血,胚水肿,肾脏出血,肝脏有坏死灶,心肌苍白。琼扩试验结果显示法氏囊样品孔、绒毛尿囊膜样品孔与标准血清孔间均可出现明显的沉淀线,说明样品中含有传染性法氏囊病病毒。根据实验室检查情况,诊断为鸡传染性法氏囊病。     

鸡传染性喉气管炎病毒培养特性的研究

本试验从发病鸡群分离一株鸡传染性喉气管炎病毒(命名为NM98a)，并用分离株和ILT王岗株对鸡胚绒毛尿囊膜、鸡胚尿囊腔及鸡胚肾细胞上培养进行比较研究。结果表明：ILTV经不同途径接种鸡胚和鸡胚肾细胞上培养，在鸡胚绒毛尿囊膜和鸡胚肾细胞上引起的病变不同，但最终增殖达到的感染性病毒量没有明显区别。鸡胚尿囊腔接种是ILTV增殖最佳途径。     

鸡传染性喉气管炎活疫苗生产工艺的改进——鸡胚绒毛尿囊膜渗透法接种应用实验

采用鸡胚绒毛尿囊膜渗透法给鸡胚接种病毒,收获感染的鸡胚绒毛尿囊膜制备鸡传染性喉气管炎活疫苗,相比于传统的人工气室绒毛尿囊膜接种法和尿囊腔接种法可降低鸡胚的早死亡率,提高鸡胚利用率,降低生产成本,提高单产量。     

鸡传染性喉气管炎弱毒活疫苗生产工艺的优化及改进试验

《中华人民共和国兽用生物制品规程》规定,在生产鸡传染性喉气管炎弱毒活疫苗时,采用鸡胚尿囊腔接种和人工气室绒毛尿囊膜接种工艺生产成品。由于人工气室绒毛尿囊膜接种早死胚多,而且工艺比较繁琐,所以我国大多数生产企业在生产中多采用尿囊腔接种方法,但尿囊腔接种后病毒繁殖时间要比人工气室绒毛尿囊膜接种后延长1~2 d,增加了孵化成本,而且绒毛尿囊膜病变(增厚、水肿等)也不如人工气室绒毛尿囊膜接种后产生的病变严重(膜重)。为此,借鉴了鸡痘鹌鹑化弱毒活疫苗的新生产工艺———鸡胚绒毛尿囊膜渗透法[1],并将此方法用于鸡传染性喉气管炎…     

肉鸡病毒性关节炎的诊断与防制

1病原 肉鸡病毒性关节炎是由呼肠孤病毒引起的传染病。病原为禽呼肠孤病毒,病毒可接种于鸡胚进行培养,病变的腱鞘经常规处理后经卵黄囊接种于5-7日龄的鸡胚,经5-6天后鸡胚死亡,胚体充血和出血,绒毛尿囊膜混浊,脾脏肿大,心肌有坏死灶。绒毛尿囊膜接种病毒时在其绒毛尿囊膜上形成大小不等隆起的疱斑。1日龄肉用雏鸡足垫内接种,第二天即可引起局部红肿等典型的病毒性关节炎。     

应用鸡胚培养试验检验鸡病毒病(Ⅲ)

4.5 鸡传染性支气管炎病料呼吸道分泌物,有病变的气管、肺。制成5—10倍悬液。接种途径经绒毛尿囊膜接种8—10日龄鸡胚。培养结果检查即便病料中含有鸡传染性支气管炎病毒,初次接种鸡胚时,亦缺乏显著病痕。然而,当用接种鸡胚尿囊液,经鸡胚连续继代2～3次后,再接种鸡胚时,便可出现本病特征性病变。鸡胚的死亡率,随继代次数的增加而增高,死亡时间亦随着提前。本病在鸡胚的特征性病变为,出现僵胚(侏儒胚)其大小有时仅为正常胚的     

鸡痘病毒FJ01株的分离鉴定

为调查福建规模化养鸡场病鸡的死亡原因,试验以福建省送检的鸡冠及头部皮肤有大量结节状痘痂的病鸡病料为研究对象进行了PCR鉴定。初步鉴定为鸡痘病毒(FWPV)株后,通过接种鸡胚绒毛尿囊膜(CAM)分离病毒;用原代鸡胚成纤维细胞(CEF)和传代细胞DF-1细胞对病毒进行传代,观察该分离毒株在两种细胞上培养特性的异同;对被感染的CEF进行超薄切片观察病毒的分布及病毒粒子的形态;并针对该分离株的TK基因和FPV175基因序列进行同源性分析。结果显示,接种经抗生素处理后的病料匀浆液上清的CAM出现大面积单个白色隆起的痘斑,匀浆痘斑后同时接种CEF和DF-1细胞,两种细胞均能产生稳定可持续传代的细胞病变,但病变出现的时间及病变程度不同;透射电镜下观察到典型的FWPV粒子密集分布在CEF的胞浆中,清晰可见卵圆形外膜包裹着的两侧凹陷的核心。对其中23个病毒粒子进行统计测得病毒粒子的大小为(258～344) nm×(153～238) nm;针对FWPV TK基因和FPV175基因的PCR检测及测序结果与GenBank收录的FWPV(登录号:NC_002188.1)核苷酸序列同源性分别高达100%和99.8%。以上结果表明该分离毒株为FWPV,命名为FWPV-FJ01,为国内FWPV的防治提供了参考依据。     

鸡传染性法氏囊炎活疫苗(B87株)生产工艺的改进--鸡胚绒毛尿囊膜渗透接种法应用试验

采用绒毛尿囊膜渗透法给鸡胚接种病毒,收获感染的胎儿及绒毛尿囊膜,制备鸡传染性法氏囊炎（B87株）活疫苗,相比于传统的人工气室绒毛尿囊膜接种法和尿囊腔接种法,可降低鸡胚早死率且无存活鸡胚,提高了鸡胚利用率,降低了生产成本.     

不同青贮玉米品种抗病性评价

对10个青贮玉米品种自然发病情况进行调查,并采用病级分类方法对抗病性分类。结果表明:对锈病表现抗性和中抗的品种有3个和5个,分别占供试品种的30%和50%;对大斑病表现抗性和中抗的品种有4个和5个,分别占供试品种的40%和50%;对褐斑病表现抗性和中抗的品种有6个和3个,分别占供试品种的60%和30%;对小斑病表现抗性和中抗的品种有6个和3个,分别占供试品种的60%和30%。多数品种对青贮玉米4种病害抗性表现较好,可从青贮玉米供试材料中选用优良抗病材料用于四川地区种植。     

Evaluation of Oxygen-dependent Immunodefences of the Polymorphonuclear Cells of Some Tropical Ruminants

Activation of polymorphonuclear cells (PMNs) leads to the formation of superoxide, which is in turn dismutated to H₂O₂ by superoxide dismutase (SOD) and is partly responsible for oxygen-dependent microbicidal activity. However, no comparative information is available on the effect of SOD inhibition before PMN activation to allow simulation of the SOD defects that are known to occur in some ruminants. This paper attempts to examine the degranulative and phagocytic responses in buffalo, cattle and goat PMNs exposed to diethyldithiocarbamate, a known SOD inhibitor. The activity of glutathione peroxidase and reductase was increased in the presence of SOD inhibitor. On activation, H₂O₂ production increased significantly (p<0.01), while SOD inhibition before the activation of PMNs caused a significant decline in the production of H₂O₂ (p<0.05) in all the species studied. There was a significant increase (p<0.05) in the phagocytosis of Candida albicans spores by buffalo PMNs activated with opsonized zymosan. Activation of bovine PMNs after exposure to the SOD inhibitor resulted in a significant decline (p<0.05) in phagocytic activity; in the other species, the two values only approached significance. Among the activators, opsonized zymosan caused a significant increase in phagocytic activity as compared to lipopolysaccharide, particularly in the PMNs of buffaloes (p<0.05). Increased fungicidal activity (p<0.05) occurred with opsonized zymosan-activated PMNs of all the species studied. The fungicidal activity was found to decline in PMNs exposed to SOD inhibitor before activation (p<0.05). Interestingly, the phagocytic activity of caprine PMNs was found to be lower than that of PMNs from cattle (p<0.05).     

Effect of dietary stable isotopic ratios of carbon and nitrogen on the extent of their incorporation into tissues of rats

This study was conducted to investigate the effect of different dietary ratios of ¹³ C to ¹² C or ¹⁵ N to ¹⁴ N on their relative incorporation into tissues. Eighty male rats were used in two 21-day feeding trials in which they were fed diets with either high δ¹³C levels (δ¹³C = −13.89‰ and δ¹⁵N = 2.37‰ in experiment 1 and δ¹³C = −19.34‰ and δ¹⁵N = 4.73‰ in experiment 2) or low δ¹³C levels (δ¹³C = −17.90‰ and δ¹⁵N = 3.08‰ in experiment 1 and δ¹³C = −21.76‰ and δ¹⁵N = 0.53‰ in experiment 2), meanwhile, the dietary δ¹⁵N levels were designed to two ranks. Blood, liver, adipose and muscle tissues were collected on day 0, 3, 7, 14, and 21 for determination of ¹³ C, ¹² C, ¹⁵ N and ¹⁴ N isotopes. Rat growth rate, antioxidant capacity and metabolic parameters were also assessed. The results indicate that adipose tissue tend to deplete ¹³ C before the stable isotopic ratios achieved final equilibrium. Therefore, feeds with different isotopic signatures had different incorporation rates into tissues. Low dietary ¹³ C levels decreased tissue δ¹³C values whereas high dietary ¹³ C levels did not alter tissue δ¹³C values during the 21-d experiment. Blood δ¹⁵N values were a reliable parameter in assessing the relative contribution of dietary nitrogen to tissues. This study revealed a relationship between dietary isotopic signatures and their incorporation rates into rat tissues. However, more studies are needed to illustrate the mechanism through which dietary isotopic ratios influence the extent of isotopic incorporation into the tissues.     

Pharmacokinetics of three formulations of vitacoxib in horses

The pharmacokinetic properties of three formulations of vitacoxib were investigated in horses. To describe plasma concentrations and characterize the pharmacokinetics, 6 healthy adult Chinese Mongolian horses were administered a single dose of 0.1 mg/kg bodyweight intravenous (i.v.), oral paste, or oral tablet vitacoxib in a 3-way, randomized, parallel design. Blood samples were collected prior to and at various times up to 72 hr postadministration. Plasma vitacoxib concentrations were quantified using UPLC-MS/MS, and pharmacokinetic parameters were calculated using noncompartmental analysis. No complications resulting from the vitacoxib administration were noted on subsequent administrations, and all procedures were tolerated well by the horses throughout the study. The elimination half-life (T_1/2λz) was 4.24 ± 1.98 hr (i.v.), 8.77 ± 0.91 hr (oral paste), and 8.12 ± 4.24 hr (oral tablet), respectively. Maximum plasma concentration (C_max) was 28.61 ± 9.29 ng/ml (oral paste) and 19.64 ± 9.26 ng/ml (oral tablet), respectively. Area under the concentration-versus-time curve (AUC_last) was 336 ± 229 ng hr/ml (i.v.), 221 ± 94 ng hr/ml (oral paste), and 203 ± 139 ng hr/ml, respectively. The results showed statistically significant differences between the 2 oral vitacoxib groups in T_max value. T_1/2λz (hr), AUC_last (ng hr/ml), and MRT (hr) were significantly different between i.v. and oral groups. The longer half-life observed following oral administration was consistent with the flip-flop phenomenon.     

Determination of optimal dietary selenium levels by full expression of selenoproteins in various tissues of broilers from 1 to 21 d of age

The current NRC dietary selenium (Se) requirement (0.15 mg/kg) of broilers is primarily based on growth performance data reported in 1986. Our study aimed to determine optimal dietary Se levels of broilers fed a practical corn-soybean meal diet for the full expression of selenoproteins in various tissues. A total of 384 one-d-old male broilers (n = 8 replicates/diet) were fed a basal corn-soybean meal diet or the basal diet supplemented with 0.1, 0.2, 0.3, 0.4 or 0.5 mg Se/kg in the form of Na₂SeO₃ for 21 d. Regression analysis was conducted to evaluate the optimal dietary Se levels using broken-line, quadratic or asymptotic models. The activity of glutathione peroxidase (GPX) in the plasma, liver, kidney and pancreas, iodothyronine deiodinase (DIO) in the plasma, liver and pancreas, and thioredoxin reductase (Txnrd) in the liver and pancreas, the mRNA levels of Gpx1, Gpx4, Dio1, selenoprotein (Seleno) h, Selenop and Selenou in the liver, Gpx4, Dio1, Txnrd1, Txnrd2, Selenoh, Selenop and Selenou in the kidney, and Gpx1, Gpx4, Selenoh and Selenou in the pancreas, and the protein levels of GPX4 in the liver and kidney of broilers were influenced (P < 0.05) by added Se levels, and increased quadratically (P < 0.05) with the increase of added Se levels. The estimates of optimal dietary Se levels were 0.07 to 0.36 mg/kg based on the fitted broken-line, quadratic or asymptotic models (P < 0.001) of the aforementioned selenoprotein expression in the plasma, liver and kidney, and 0.09 to 0.46 mg/kg based on the fitted broken-line models (P < 0.001) of the aforementioned selenoprotein expression in the pancreas. The results indicate that the optimal dietary Se levels would be 0.36 mg/kg to support the full expression of selenoproteins in the plasma, liver and kidney, and 0.46 mg/kg to support the full expression of selenoproteins in the pancreas of broilers fed a practical corn-soybean meal diet from 1 to 21 d of age.     

Influence of fibre and betaine on development of the gastrointestinal tract of broilers between hatch and 14 d of age

An experiment was designed to determine the influence of fibre and betaine on the development of the intestine, liver and pancreas of broilers from hatch to 14 d of age. A total of 250-day-old Cobb 500 male broilers were allocated to 16 cages with 15 broilers each. Treatments were arranged in a 2 × 4 factorial design, consisting of 2 feed formulations (low and high fibre diets) and 4 levels of betaine (0, 1, 3 or 5 kg/t). At hatch, 10 birds in total were euthanised, and samples of the liver, pancreas, yolk sac and intestine were collected for reference of the analysed parameters before the start of the trial. On d 4, 9 and 14, 5 birds per cage (10 birds per treatment) were selected, euthanised and treated as the same as the birds at hatch. Villus height and width and crypt depth were determined on the duodenum samples, and absorptive area was calculated. The number of enterocytes in mitosis at the villus was determined by a positive reaction to antibody for Ki67 protein, and fused villus was evaluated visually. The relative weight of the yolk sac reduced (P < 0.05) as birds aged while the intestine and liver reached a maximum (P < 0.05) at around d 4 and the pancreas at d 9. Birds fed the high fibre diet had greater feed intake, lower relative weight of the pancreas and higher villus (P < 0.05) than birds fed the low fibre diet. Villus width increased (P < 0.05) at 4 d of age, and this was associated with fused villus. Betaine inclusion reduced (P < 0.05) villus width, increased (P < 0.05) villus size and absorptive area, and reduced (P < 0.05) the number of enterocytes with positive reaction for the antibody Ki-67. Betaine inclusion reduced the width and increased the absorptive area and the villus height of the duodenum of birds up to 14 d of age. The higher fibre diet increased feed intake and villus height, yet reduced pancreas relative weight, while not affecting body weight gain. This response was possibly due to a dilution effect of the fibre, reducing nutrient absorption and consequently stimulating villus growth to improve absorption rates.     

Effects of GnRH treatment on initiation of pulses of LH,LH release,and subsequent concentrations of progesterone

Progesterone is essential for establishment and maintenance of pregnancy. One proposed method to increase progesterone is administering GnRH at insemination. However, this method has resulted in conflicting results. Therefore, 2 experiments were conducted to evaluate how administering GnRH at insemination affected pulses of luteinizing hormone (LH) and subsequent progesterone. In Experiment 1, cows were allotted to 2 treatments: (1) GnRH (100 μg) given approximately 12 h after initiation of estrus (n = 5); and (2) Control (n = 5). Blood samples were collected at 15-min intervals for 6 h at 12 (blood sampling period 1), 26 (blood sampling period 2), 40 (blood sampling period 3), 54 (blood sampling period 4), and 68 (blood sampling period 5) h after onset of estrus. Daily blood samples were collected for 17 d. In Experiment 2, cows were allotted into 2 treatments: GnRH administered 10 to 11 h (n = 10) or 14 to 15 h (n = 10) after onset of estrus. Daily blood samples were collected for 17 d. Cows treated with GnRH tended (P ≤ 0.075) to have greater LH release during blood sampling period 1, tended (P = 0.095) to have fewer pulses during blood sampling period 2, tended (P = 0.067) to have greater concentrations of progesterone, and had an earlier (P = 0.05) increase in progesterone than control cows. Cows treated with GnRH 10 to 11 h after onset of estrus had greater (P = 0.01) progesterone and an earlier (P = 0.04) increase in progesterone than cows treated 14 to 15 h. In conclusion, timing of GnRH treatment following onset of estrus influenced pulses of LH and subsequent progesterone.     

In Vitro Evaluation of the Effects of Selected Plants on the Growth of the Mycelial Form of Histoplasma capsulatum Variety farciminosum in Ethiopia

Epizootic lymphangitis is prevalent in equines in Ethiopia, causing remarkable economic and welfare impacts but often neglected. Lack of effective treatment contributed to its continued occurrence, and hence, search for an effective treatment should be considered a priority area to minimize its impacts. Previous ethnobotanical studies have reported that Curcuma longa, Phytolacca dodecandra, and Datura stramonium were used to treat cutaneous fungal infections and reduce their incidence. The treatment effects of these plants against epizootic lymphangitis should be studied. The in vitro growth inhibitory effects of methanol extracts of the root of C. longa, berry of P. dodecandra, and leaf of D. stramonium were evaluated. Histoplasma capsulatum var farciminosum was isolated from clinical cases of epizootic lymphangitis in carthorses in central Ethiopia. The nested polymerase chain reaction was used to confirm the identity of the isolates. Serial twofold dilutions of the extract of berries of P. dodecandra and leaves of D. stramonium were done in sterile water, whereas dilution of the extract of roots of C. longa was done in dimethylsulphoxide. The effects of the plants on the growth of Histoplasma capsulatum var farciminosum were assessed by agar dilution assay. Culture media with no antifungal agent and media containing ketoconazole served as negative and positive control, respectively. The methanol extract of C. longa showed inhibitory effects at concentrations ranging from 0.07 to 5 mg/mL. Similarly, the methanol extract of P. dodecandra showed growth inhibitory effects at concentrations ranging from 0.156 to 5 mg/mL. That is, the growth inhibitory concentration of C. longa was 0.07 mg/mL, whereas that of P. dodecandra was 0.156 mg/mL. In contrast, D. stramonium showed no inhibitory effect. This preliminary observation showed that methanol extracts of C. longa and P. dodecandra showed inhibitory effects on the growth of Histoplasma capsulatum var farciminosum requiring further repeated in vitro evaluation so as to generate adequate evidence, which would justify in vivo trials.     

Effects of the ruminal comminution rate and microbial contamination of particles on accuracy of in situ estimates of ruminal degradability and intestinal digestibility of feedstuffs

Effects of considering the comminution rate (k_c) and the correction of microbial contamination (using ¹⁵N techniques) of particles in the rumen on estimates of ruminally undegraded fractions and their intestinal digestibility were examined generating composite samples (from rumen‐incubated residues) representative of the undegraded feed rumen outflow. The study used sunflower meal (SFM) and Italian ryegrass hay (RGH) and three rumen and duodenum cannulated wethers fed with a 40:60 RGH to concentrate diet (75 g DM/kgBW^0.75). Transit studies up to the duodenum with Yb‐SFM and Eu‐RGH marked samples showed higher k_c values (/h) in SFM than in RGH (0.577 vs. 0.0892, p = 0.034), whereas similar values occurred for the rumen passage rate (k_p). Estimates of ruminally undegraded and intestinal digestibility of all tested fractions decreased when k_c was considered and also applying microbial correction. Thus, microbial uncorrected k_p‐based proportions of intestinal digested undegraded crude protein overestimated those corrected and k_c?k_p‐based by 39% in SFM (0.146 vs. 0.105) and 761% in RGH (0.373 vs. 0.0433). Results show that both k_c and microbial contamination correction should be considered to obtain accurate in situ estimates in grasses, whereas in protein concentrates not considering k_c is an important source of error.     

Effects of oral supplementation of probiotic strains of Lactobacillus rhamnosus and Enterococcus faecium on diarrhoea events of foals in their first weeks of life

Foal first diarrhoea is one of the most prominent problems in the early life of horses. Probiotics might have the potency to prevent or at least diminish neonatal diarrhoea. We hypothesised that the treatment of foals with probiotic strains of Lactobacillus rhamnosus and Enterococcus faecium starting early after birth and then daily over 2 weeks would prevent or mitigate foal heat diarrhoea. The influence of this probiotic treatment on diarrhoea incidence and growth and health performance of young foals was investigated. Thirty‐four foals were randomly allocated to two groups. From day 1 to 14 of life, the foals received either placebo (PG, n = 16) or the probiotic treatment (TG, n = 18). Clinical examination was performed, and the faeces consistency score (FCS, 1–5; with diarrhoea defined by ≤3) was recorded once per day in weeks 1 and 2 and once weekly in weeks 3–8 of life (WL). The body height was measured at birth and after two and eight WL. Diarrhoea occurred in the 1st WL in 19% and 61% of PG and TG foals respectively. In the 1st WL, diarrhoea lasted 0.3 ± 0.8 and 1.6 ± 1.4 days in PG and TG foals respectively. In the 2nd WL, diarrhoea occurred in 94% and 84% of PG and TG foals, respectively, and lasted for 3.0 ± 1.5 and 3.7 ± 1.6 days respectively. At least two periods of diarrhoea developed in 33% and 65% of PG and TG foals respectively. The TG foals grew slightly slower than the PG foals. The results indicated that the probiotic treatment of neonatal foals as performed in this study was not suitable to reduce diarrhoea within the first two WL, because contrary to the hypothesis, the TG foals suffered more frequently and for longer periods from diarrhoea than the PG foals.