Effect of vaccination and the immunomodulators “bacillus of Calmette–Guérin, Avridine and Propionibacterium acnes” on rabies in mice

Responses of vaccination and treatment to immunomodulators against rabies in mice were evaluated through macrophage inhibition factor (MIF), intra-pad inoculation (IPI) and serum neutralization (SN) tests and by the detection of γ-interferon (IFN-γ). Onco-BCG, Avridine and Propionibacterium acnes were administered to groups of mice. Higher survival rates were found in animals treated with P. acnes. Lower levels of IFN-γ were observed in the groups of infected and vaccinated mice. The IPI was not effective on detecting the response of delayed-type hypersensitivity. Vaccine induced in the infected animals a more intense response to MIF reaction.

Résumé

Les réponses aux immunomodulateurs vis à vis de la vaccination et du traitement antirabique chez la souris ont été évaluées à travers le facteur inhibant les macrophages, inoculation du test du coussinet plantaire et des anticorps neutralisants ainsi que la détection d'interféron γ. Onco-BCG, Avridine et Propionibacterium acnes ont été administrés à des groupes de souris. Le taux de survie le plus élevé a été observé chez les animaux traités par P. acnes. Les taux les plus bas d'interféron ont été observés dans les groupes de souris infectées et vaccinées. Le test du coussinet plantaire n'a pas permis de détecter la réponse d'hypersensibilité retardée. Le vaccin induit chez les animaux infectés une réponse plus intense au facteur inhibant les macrophages.     

Evaluation of cytokines concentration and percentage of survival of rabies virus-infected mice submitted to anti-rabies Vero-cell propagated vaccine and P. acnes

Previously, survival of rabies infection was shown to correlate with low IL-6 serum concentration in mice subjected to post-exposure treatment with the Fuenzalida Palacios rabies vaccine in conjunction with the immunomodulator Propionibacterium acnes, previously Corynebacterium parvum. Considering the substitution of the Fuenzalida Palacios rabies vaccine by the Vero cell raised anti-rabies vaccine in almost all countries, the objective of this work was to evaluate the survival and cytokine serum concentration of rabies virus-infected mice treated with P. acnes in conjunction with or the anti-rabies-VERO vaccine. For this, Swiss mice were experimentally infected with street rabies virus and subjected to vaccine and/or P. acnes following infection. Animals were killed at different times and serum was collected to evaluate cytokines. The greatest survival was observed in animals given one or two does of P. acnes in the absence of vaccination. Animals given anti-rabies VERO vaccine alone or with three doses of P. acnes had the second highest survival rate. The group that had the highest percentage of mortality also had the highest IL-6 concentration on the 10th day, a time correlating with clinical symptoms of the animals. The results reinforce the inefficacy of anti-rabies vaccine in only one dose as a post-exposure treatment irrespective of the type of vaccine used, the immunomodulation activity of P. acnes in rabies post-exposure treatment and suggest a role for IL-6 in rabies virus pathogenesis.     

伯氏疟原虫ANKA株感染小鼠的T细胞、NK细胞及细胞因子变化

旨在系统分析伯氏疟原虫感染引起宿主T细胞、NK细胞、Tim-3表达及细胞因子的变化。选取64只雌性C57BL/6小鼠随机分为8组,每组8只,分别于感染后0、4、7、9、11、13、16和19 d获取小鼠脾及外周血免疫细胞,利用流式细胞术检测小鼠主要免疫细胞亚群及免疫检查点分子Tim-3表达水平的变化;同时检测血清中细胞因子的变化。结果表明,感染疟原虫后,小鼠脾CD3⁺CD4⁺ T细胞、CD3⁺CD8⁺ T细胞及NK细胞的比例均逐渐降低（P<0.01）,且伴随着3种细胞Tim-3表达量的升高（P<0.01）。小鼠外周血CD3⁺CD4⁺ T细胞的比例呈先降低后升高趋势,CD3⁺CD8⁺ T细胞的比例呈先升高后降低趋势（P<0.05）;小鼠外周血CD3^-NK1.1⁺细胞的比例呈先降低后升高趋势,但感染末期,其比例仍低于未感染组（P<0.05）。Tim-3分子在外周血CD3⁺CD4⁺ T细胞、CD3⁺CD8⁺ T细胞及CD3^-NK1.1⁺细胞的表达均显著升高（P<0.05）。感染疟原虫后,小鼠血清中促炎细胞因子IL-2的分泌量均显著高于未感染组（P<0.05）;促炎细胞因子IFN-γ、TNF-α和IL-6在血清中分泌均呈先升高后降低趋势（P<0.05）;具有免疫抑制作用的细胞因子IL-10呈逐渐升高趋势,且感染后期急剧升高（P<0.001）。以上结果表明,感染疟原虫后,小鼠的特异性免疫反应发挥了一定的杀伤作用,但由于Tim-3免疫检查点分子及一些发挥免疫抑制作用的细胞因子（IL-10）的过度表达,有利于疟原虫逃避宿主的免疫捕杀作用。该研究提示了从宿主免疫抑制角度研究疟原虫感染的重要性。     

Humoral and cellular immune response of rabbits to Psoroptes cuniculi, the rabbit scab mite.

Rabbits infested for the first time with Psoroptes cuniculi (Group A) and heavily infested ones with unkown aetiology (Group B) were examined for specific serum antibody activity and responsiveness of their peripheral blood lymphocytes by the enzyme-linked immunosorbent assay and lymphocyte transformation assay methods. These parameters were examined after treatment with ivetmectin and after subsequent challenge infestation with P. cuniculi. Group A rabbits developed a small number of mite-caused lesions, and exhibited a significant P. cuniculi antigen-induced T cell response and a high level were observed to be suppressed in Group B rabbits that were highly susceptible to P. cuniculi.     

饲粮中添加牛至精油和莫能菌素对荷斯坦犊牛血清生化指标、消化酶活性及瘤胃微生物区系的影响

旨在研究牛至精油（oregano essential oil,OEO）与莫能菌素（monensin,MON）对荷斯坦犊牛血清生化指标、消化酶活性以及瘤胃微生物区系的影响。本试验前期阶段选用了体重相近（（41.31±2.23）kg）,体况良好的荷斯坦初生犊牛80头,随机分为4组,每组20头,采用单栏饲养。对照组饲喂全脂牛乳和颗粒饲料,其他3个试验组在饲喂全脂牛乳和颗粒饲料的基础上分别添加4 g·kg^-1牛至精油、3.6 g·kg^-1莫能菌素和7.6 g·kg^-1牛至精油和莫能菌素的混合物,饲喂至56、70日龄时,从每组中选取6头体重相近的公犊,颈静脉采血测定血清指标,并通过口腔导管的形式采集犊牛的瘤胃液,检测消化酶活性以及瘤胃微生物区系。结果表明：1）添加牛至精油显著提高了56日龄犊牛血清中TP、SOD、GSH和GSH-Px的浓度（P ≤ 0.05）;莫能菌素对56、70日龄犊牛BUN浓度有降低的趋势（P<0.10）。2）牛至精油显著降低了56、70日龄犊牛瘤胃中产琥珀酸丝状杆菌的数量（P<0.05）,显著提高了白色瘤胃球菌的数量（P ≤ 0.05）;莫能菌素显著降低了56、70日龄犊牛瘤胃中原虫的数量（P ≤ 0.05）,并有降低白色瘤胃球菌数量的趋势（P<0.10）,在56日龄时,莫能菌素有降低犊牛瘤胃中黄色瘤胃球菌数量的趋势（P<0.10）;牛至精油和莫能菌素对56、70日龄犊牛瘤胃中溶纤维丁酸弧菌有显著的交互作用（P ≤ 0.05）;在56日龄时,牛至精油和莫能菌素对栖瘤胃普雷沃氏菌有交互作用的趋势（P<0.10）,而在70日龄时则呈显著的交互作用（P ≤ 0.05）。3）牛至精油和莫能菌素对56、70日龄犊牛瘤胃中胃蛋白酶浓度有显著的交互作用（P ≤ 0.05）。综上所述,饲粮中添加牛至精油可提高犊牛的免疫力和抗氧化能力。莫能菌素在抑制瘤胃内原虫数量上强于牛至精油,牛至精油对提高主要蛋白质降解菌数量的作用效果优于莫能菌素。总之,牛至精油可改善犊牛的健康状况以及调控瘤胃微生物区系,牛至精油作为天然植物抗生素可替代犊牛期预防性治疗的其他类抗生素生长促进剂。     

Dietary supplementation with Chinese herbal ultra-fine powder enhances cellular and humoral immunity in early-weaned piglets

This study was conducted to determine effects of dietary supplementation with a Chinese herbal ultra-fine (CHU) powder on cellular and humoral immunity in early-weaned piglets. Sixty piglets weaned at 21 days of age were randomly assigned to one of 3 treatment groups representing supplementation with 0 or 2 g/kg of the powder, or 0.2 g/kg of colistin (an antibiotic) to maize- and soybean meal-based diets (n = 20). On days 7, 14 and 28 after initiation of the treatment, lymphocyte proliferating activity and serum levels of antibodies and cytokines were measured. Results indicated that the powder enhanced (P < 0.05) serum concentrations of IL-2, IL-6 and TNF- on days 7 and 28, and of IgG and IgM on day 28, as well as the proliferating activity of blood lymphocytes in vivo on all days relative to the none additive group. Addition of the powder (7.82 to 31.25 mg/ml) to culture medium in vitro also increased (P < 0.05) the proliferating activity of blood lymphocytes compared with the none additive group. These findings indicate a beneficial effect of the CHU powder on cellular and humoral immune responses in piglets.     

高寒矿区渣山坡向对恢复植物叶片养分重吸收的影响

植物叶片养分重吸收是植物适应生境胁迫的重要机制。本试验以高寒矿区渣山阳坡、山顶平地和阴坡上播种的恢复植物老芒麦（Elymus sibiricus）、冷地早熟禾（Poa crymophila）和小花碱茅（Puccinellia tenuiflora）生长旺盛期的成熟叶片及凋亡期的衰落叶片为研究对象,测定不同坡向,不同时期的叶片氮磷含量,研究坡向对恢复植物叶片氮、磷重吸收效率的影响。结果表明：从阳坡、平地到阴坡,恢复植物成熟叶片养分含量以及养分重吸收效率逐渐降低;阳坡上老芒麦,平地上冷地早熟禾以及阴坡上小花碱茅的衰落叶片氮含量较低而氮素重吸收效率较高;恢复植物氮素重吸收效率均显著高于磷素重吸收效率,且此效应在阴坡上显著高于阳坡和平地。在高寒矿区渣山恢复中,应依据坡向差异选择养分重吸收效率较高的恢复物种,如阳坡上适宜播种老芒麦和冷地早熟禾,平地上适宜播种冷地早熟禾,阴坡上适宜播种小花碱茅,有助于提升渣山受损生态系统的恢复效率。     

钙盐胁迫对3份葛藤种质种子萌发及幼苗生理特性的影响

为探明葛藤对喀斯特山区土壤富钙环境的适应机制,以3份不同产地葛藤种质种子为试验材料,采用室内模拟试验,研究不同浓度（0、50、100、150和200 mmol·L^-1 CaCl₂）钙盐胁迫对3份葛藤种质种子萌发、幼苗生长、渗透调节物质、抗氧化酶系统及植物内源激素的影响。结果表明,随钙盐胁迫浓度升高,澳大利亚（AUS）和江苏（JS）种质种子发芽率、发芽势、发芽指数、活力指数均显著降低,种子萌发率、萌发速度及种苗生活力均受到显著抑制（P<0.05）,湖南（HN）种质在胁迫浓度为100 mmol·L^-1时,种子发芽率、发芽势、发芽指数升高;3份葛藤种质幼苗叶长、叶宽、株高和生物量显著降低;叶片丙二醛（MDA）含量和相对电导率均显著升高;叶片脯氨酸（Pro）、可溶性糖含量和超氧化物歧化酶（SOD）、过氧化氢酶（CAT）活性均先增后降;AUS和HN种质叶片过氧化物酶（POD）活性先增后降,JS种质表现为显著上升;HN和JS种质叶片抗坏血酸过氧化物酶（APX）活性显著升高,AUS种质则表现为先升后降;3份葛藤种质叶片脱落酸（ABA）、生长素（IAA）、赤霉素（GA₃）、玉米素（ZT）（除JS种质）含量均先增后减。0~50 mmol·L^-1 CaCl₂胁迫对3份葛藤种质种子萌发及幼苗生长抑制不显著,150~200 mmol·L^-1 钙盐胁迫显著抑制3种葛藤种子萌发及幼苗生长,HN种质种子依靠种子休眠抵御钙盐胁迫,说明葛藤幼苗对低钙盐胁迫具有一定的耐受性。中高钙盐浓度胁迫时,葛藤幼苗通过提高自身渗透物质含量、增强抗氧化酶活性、增加部分激素含量、改变地上地下生物量等方式积极调节自身生理代谢, 以适应高钙盐环境。结合隶属函数和主成分分析,除Pro含量和APX活性外,其余指标均可作为评价3份葛藤种质耐盐性的主要指标,3份种质的耐盐性大小顺序为JS>HN>AUS。     

灌服黄芪多糖对奶牛产奶量、免疫性能及抗氧化性能的影响

【目的】研究黄芪多糖(APS)对奶牛产奶量、免疫功能及抗氧化性能的影响。【方法】选取体况良好,体重、产奶量、泌乳天数、胎次接近的中国荷斯坦奶牛44头,随机分为对照组(CK)及APS20、APS55和APS90组,每组11头。所有试验牛均饲喂基础饲粮。对照组灌服温水200 mL,试验组分别灌服20、55、和90 g/d的APS溶液200 mL,预试期10 d,正试期60 d。正试期内,每10 d统计1次产奶量;于正试期的第0、30、60天采集乳样,测定乳中体细胞数(SCC);于正试期第0、30、60天晨饲前,采集奶牛尾根静脉血样,测定血清中相关免疫指标、抗氧化指标。【结果】在试验全期,各组产奶量均差异不显著(P＞0.05)。在体细胞数方面,与CK组相比,第30和60天时,APS55组体细胞评分(SCS)均显著降低(P＜0.05),APS20和APS90组SCS在各时间点均差异不显著(P＞0.05),且APS90组极显著高于APS55组(P＜0.01)。在免疫指标方面,与CK组相比,第30天时,APS20组血清中白细胞介素2(IL-2)含量显著增加(P＜0.05),APS55与APS90组血清中IL-2含量极显著增加(P＜0.01);第60天时,APS20、APS55和APS90组血清中IL-2含量均极显著增加(P＜0.01),且APS55与APS90组IL-2含量极显著高于APS20组(P＜0.01);第60天时,APS55组血清中IL-6含量显著增加(P＜0.05)。在抗氧化指标方面,与CK组相比,第30、60天时,APS20、APS55和APS90组血清中过氧化氢酶(CAT)活性均极显著增加(P＜0.01);第60天时,ASP90组血清中丙二醛(MDA)浓度显著降低(P＜0.05),APS20、APS55和APS90组血清中谷胱甘肽过氧化物酶(GSH-Px)活性均显著增加(P＜0.05),APS55和APS90组血清中总抗氧化能力(T-AOC)极显著增加(P＜0.01),且显著高于APS20组(P＜0.05)。【结论】55 g/d APS可以提高奶牛免疫功能、抗氧化能力,并且降低乳中体细胞数,结果为APS在奶牛生产应用提供参考。     

9个野生早熟禾对低温胁迫的生理响应及苗期抗寒性评价

以甘肃省境内采集的9个野生早熟禾（Poa）为试验材料,设5,0,-5,-10℃共4个温度处理,探讨其对低温胁迫的生理响应和抗寒性,为抗寒早熟禾种质材料筛选和新品种选育提供依据。结果表明：随着温度下降,各材料的叶片相对膜透性逐渐增大,游离脯氨酸（Pro）、可溶性糖（SS）、可溶性蛋白（SP）、丙二醛（MDA）含量逐渐增加,超氧化物歧化酶（SOD）和过氧化物酶（POD）活性逐渐增强。运用隶属函数法综合评价9个材料苗期抗寒性发现,一年生早熟禾（P. annua）抗寒性最强,小药早熟禾（P. micrandra）、草地早熟禾（天祝）（P. pratensis）和草地早熟禾（甘南）抗寒性中等;硬质早熟禾（P. sphondylodes）、波伐早熟禾（P. poophagorum）、草地早熟禾（肃南）、草地早熟禾 （渭源）、草地早熟禾（兴隆山）抗寒性弱。相关性分析表明,SP含量与抗寒综合评价值极显著正相关（P<0.01）,POD活性与抗寒综合评价值显著正相关 （P<0.05）,Pro含量、SS含量、SOD活性、海拔与抗寒综合评价值成正相关,相对膜透性、MDA含量和年均温与抗寒综合评价值成负相关。     

辣蓼黄酮乙酸乙酯部分对PRV体外诱导RAW264.7细胞分泌炎性因子的影响

探讨辣蓼黄酮乙酸乙酯部分（ethyl acetate of flavonoids from Polygonum hydropiper L.,FEA）对猪伪狂犬病病毒（Pseudorabies virus,PRV）体外诱导RAW264.7细胞分泌炎性因子的影响。正式试验前将不同浓度的FEA作用于RAW264.7细胞,通过CCK-8检测细胞体外增殖活性,筛选FEA对RAW264.7细胞的安全浓度范围。正式试验分为空白对照组、PRV阳性对照组、芦丁对照组、5个FEA药物组,除空白对照组外,其余各组细胞加入PRV共孵育1.5 h后,再加入芦丁或不同浓度的FEA,分别培养4、8、12、24 h,CCK-8法检测FEA对病毒感染的RAW264.7细胞活性,筛选出3个FEA药物组（高、中、低剂量FEA）,再通过ELISA法检测各时间点细胞培养液上清中炎症相关因子TNF-α、IL-1β、IL-6、IL-10、MCP-1和IFN-γ的分泌水平,以确定FEA体外调节炎症反应的最适时间及药物浓度。结果显示：①FEA对RAW264.7细胞的安全浓度为12.5~200 μg/mL;②25~100 μg/mL FEA能显著提高PRV感染的RAW264.7细胞体外增殖活性;③PRV感染RAW264.7细胞后促进细胞炎症因子的分泌,用FEA处理8~12 h后,在一定程度上降低了TNF-α、IL-1β、IL-6和MCP-1的分泌水平（P<0.05）,升高了IFN-γ分泌水平（P<0.05）,调节了IL-10分泌水平,且FEA处理8 h抗炎效果最好。结果表明,FEA能调节病毒感染细胞分泌炎性因子的水平,提示其具有体外抗炎作用。该结果可为进一步研究FEA抗病毒分子机制提供参考依据。     

Release of tumor necrosis factor-alpha from bovine alveolar macrophages stimulated with bovine respiratory viruses and bacterial endotoxins

The release of tumor necrosis factor-alpha (TNF-) from cultured bovine alveolar macrophages (BAM) was evaluated following stimulation of BAM with bovine herpesvirus-1 (BHV-1), parainfluenza-3 (PI-3) virus, bovine respiratory syncytial virus (BRSV), Escherichia coli 0111:B4 endotoxin, Pasteurella haemolytica type 1 endotoxin, Pasteurella multocida endotoxin, and virus/endotoxin combinations. A cytotoxic assay system using Georgia bovine kidney cells as targets was used to measure TNF- activity. The cytotoxic activity was neutralized by an anti-human TNF- monoclonal antibody.

Stimulation of BAM with 1 median tissue culture infectious dose (TCID₅₀) of live or ultraviolet (UV)-inactivated PI-3 virus/cell resulted in release of TNF- in significantly (P<0.05) higher amounts than sham-induced BAM. The quantities of TNF- released after live or UV-inactivated BHV-1 or BRSV induction were not significantly higher than sham-induced BAM. E. coli 0111:B4, P. haemolytica type 1 and P. multocida endotoxins stimulated TNF- release in a dose-dependent manner. Sequential exposure of BAM to 1 TCID₅₀ per cell of either live BHV-1, PI-3 virus or BRSV and then 5 μg ml⁻¹ of either E. coli 0111:B4, P. haemolytica type 1 or P. multocida endotoxin caused a significant (P<0.05) reduction in detectable TNF- in seven of nine virus/endotoxin combinations tested, when compared with 5 μg ml⁻¹ of endotoxin alone. Parainfluenza-3 virus/endotoxin combinations stimulated higher TNF- release when compared with other virus/endotoxin combinations. Five out of six test animals had serum-neutralizing antibodies to PI-3 virus, one out of six had serum-neutralizing antibodies to BHV-1, and two out of six had serum-neutralizing antibodies to BRSV, suggesting a possible relationship between serum neutralizing antibodies and TNF- release from in vitro cultivated BAM.     

Pathophysiological and immune cell responses in calves prior to and following lung challenge with formalin-killed Pasteurella multocida biotype A:3 and protection studies involving subsequent homologous live challenge

Pneumonic pasteurellosis is a common respiratory infection in cattle that has major economic and welfare implications world-wide and the incidence in the UK due to Pasteurella multocida, currently the same as that associated with Mannheimia haemolytica, is increasing. Whereas much is known regarding the pathogenesis of M. haemolytica infections little information is available on the pathogenic process of pasteurellosis initiated by P. multocida. In the present work calf systemic and innate immune responses to intratracheal challenge with formalin-killed P. multocida biotype A:3 and to subsequent experimental lung infection with live P. multocida were investigated. Eight-week-old calves were challenged intratracheally on day 0 with either 10⁹ colony forming units (cfu) of formalin-killed P. multocida biotype A:3 in 300 ml saline (n=10) or 300 ml saline alone (n=10), followed, at day 21, by challenge with 10⁹ cfu live P. multocida. Pathophysiological and lung phagocyte responses were assessed by clinical monitoring, sequential lung lavage and blood sampling. Results for samples obtained before, during and after challenge showed clinical and acute phase protein responses to both bacterial culture and saline control treatments, although higher responses were associated with bacterial challenge. Phagocytosis of P. multocida during 1 h incubation periods with lavaged cells in vitro was unaffected by exposure in vivo to killed P. multocida and there was evidence that P. multocida was able to survive intracellularly during this assay. There was no indication that lung exposure to formalin-killed P. multocida conferred protection against subsequent homologous live challenge.     

UBC13对支气管哮喘模型小鼠Th2、Th17型细胞极化的影响

试验旨在探究UBC13蛋白对支气管哮喘小鼠体内Th2、Th17细胞极化的影响。18只BALB/c小鼠随机均分为3组：PBS组、哮喘模型（OVA）组和UBC13蛋白（UBC13）组,OVA组和UBC13组采用卵清蛋白（OVA）和脂多糖（LPS）进行致敏和雾化建立哮喘模型,其中UBC13组于每次雾化前0.5 h腹腔注射100 μg UBC13蛋白。末次雾化激发24 h后处死小鼠,采集样品,通过HE染色观察肺脏切片病理学变化、PAS染色观察气道黏液的分泌,ELISA法检测血清IgE浓度和肺支气管肺泡灌洗液（BALF）上清中IL-4、IL-5和IL-17A的水平,实时荧光定量PCR法检测肺脏Th2型相关因子IL-4、IL-5、IL-13、IFN-γ mRNA和Th17型细胞因子IL-1β、IL-6、IL-17A、IL-23 mRNA的相对表达量。结果显示,试验成功建立了哮喘模型,与PBS组相比,OVA组IgE浓度极显著上升（P < 0.01）,肺脏病理切片炎性细胞浸润和中性黏液分泌现象明显,肺脏中IL-1β、IL-4、IL-6、IL-13和IL-17A mRNA相对表达量均极显著升高（P < 0.01）,IL-5和IL-23 mRNA相对表达量显著升高（P < 0.05）,IFN-γ mRNA相对表达量极显著下降（P < 0.01）,BALF中IL-4、IL-5和IL-17A的水平极显著上升（P < 0.01）;与OVA组相比,UBC13组IgE浓度显著降低（P < 0.05）,病理切片炎性细胞浸润现象减轻、黏液分泌减少,肺脏IL-1β、IL-4、IL-17A、IL-13和IL-23 mRNA相对表达量均显著下降（P < 0.05）,IFN-γ mRNA相对表达量显著升高（P<0.05）;IL-6 mRNA相对表达量极显著下降（P < 0.01）,BALF中IL-5的水平极显著下降（P < 0.01）,IL-4的水平显著下降（P < 0.05）,IL-17A的水平无显著差异（P > 0.05）。由此可见,UBC13蛋白能下调Th2和Th17型细胞因子的表达量,影响哮喘模型Th2和Th17细胞的极化。     

7种优良观赏草光合生理日变化及光响应特征研究

采用便携式LI-6400光合测定仪,选择晴朗天气对观赏草劲芒、柳枝稷、花叶虉草、歌舞芒、矮蒲苇、遍生芒、晨光芒的叶片细胞间CO₂浓度(Ci)、气孔导度(Cond)、气孔限制值、净光合速率(Pn)、瞬时光合有效辐射(PAR)和蒸腾速率(Tr)等有关参数进行测定,并计算各材料的光合饱和点(LSP)和光合补偿点(LCP),研究材料光合日变化及其耐荫性,测定结果表明,7种观赏草中,柳枝稷的光补偿点、净光合速率和水分利用率(WUE)最高,抗旱耐荫。晨光芒和花叶虉草均有较高气孔导度,两者在净光合速率和蒸腾速率方面,晨光芒最高,花叶虉草最低,花叶虉草WUE最低,抗旱性差,在潮湿或水环境中表现好,为具有较强耐荫能力C₃型植物,出现“光合午休”现象,主要由气孔导度下降引起。根据测定的数据及田间不同相对光强下材料的生长情况综合评估,7份材料的耐荫程度由强到弱依次是矮蒲苇、花叶虉草、歌舞芒、遍生芒、劲芒、柳枝稷、晨光芒。     

贵州省山羊源流产细菌调查及致病机理初探

为调查贵州省细菌性山羊流产疫病的主要原因,分析山羊源流产细菌对正常怀孕动物免疫平衡的影响,本试验采集贵州省6个地区(州、市)12个规模养羊场临床健康和发病流产山羊的胎儿、子宫和母羊的阴道棉拭子样本共计107份进行细菌分离和鉴定,采用药敏纸片琼脂扩散法对3种主要羊源分离菌进行20种抗生素的耐药性调查,并用羊源流产细菌建立BALB/c小鼠流产模型,通过ELISA方法对妊娠相关细胞因子和激素进行测定。结果显示,从采集的107份样本中共分离到13个属细菌共302株,其中发病流产山羊生殖器官中分离的细菌种类多于临床健康山羊;流产山羊生殖器官中分离的羊源细菌耐药率高于临床健康山羊细菌耐药率;3种主要羊源流产细菌埃希氏菌、葡萄球菌和链球菌均可引起怀孕BALB/c小鼠发生不同程度的流产,流产率为埃希氏菌>葡萄球菌>链球菌;胚胎吸收率为埃希氏菌>葡萄球菌>链球菌;与空白对照组相比,各试验组BALB/c小鼠血清中的IFN-γ、IL-2含量上升,而IL-4、IL-10和孕酮含量降低。结果表明,贵州省细菌性山羊流产的发生与山羊生殖器官细菌种类多少、耐药性高低有一定相关性,推测山羊源流产细菌可能通过影响怀孕山羊的主要妊娠相关细胞因子和激素的含量导致山羊流产。     

Preliminary Study on Abortion Bacteria from Goat in Guizhou Province and their Pathogenic Mechanism

In order to study the main reason of bacterial goat abortion in Guizhou province,and analyze the bacteria impact of abortion goat immune balance in normal pregnancy animals,107 samples of the goat fetus,uterus and vaginal swabs were collected from clinical health and morbidity abortion goat in 12 scale goat farms of 6 areas in Guizhou province.The bacteria were isolated and identified.The drug susceptibility disc agar diffusion method was launched for the survey of the resistance of three main sources of goat isolates to 20 kinds of antibiotics.The BALB/c mice model of abortion was established using abortion bacteria isolated from goats.The pregnancy related hormones and cytokines were determined by ELISA method.The results showed that 302 strains bacteria of 13 genera were isolated from the samples,and the species of bacteria isolated from morbidity abortion goat were more than the clinical health goats.The bacteria resistant rate of goat reproductive organs in abortion goats was higher than that of clinical healthy goats bacteria.The varying degrees of abortion of BALB/c mice could cause by Escherichia,Staphylococcus and Streptococcus isolated from goats,the rates were Escherichia>Staphylococcus>Streptococcus,respectively and the embryo resorption rates were Escherichia>Staphylococcus>Streptococcus,respectively.Compared with the control group,IFN-γ and IL-2 in the serum were increased;However,IL-4,IL-10 and progesterone were decreased.The results indicated that the occurrence of bacterial goat abortion diseases had certainly related to the number of bacterial types and the level of drug resistance in goat reproductive organs.The levels of pregnancy related hormones and cytokines in goats serum could be changed by abortion bacteria,thus lead to goats abortion.     

不同粒度猫尾草对羔羊体外发酵特性和微生物数量的影响

通过Rusitec-s体外发酵系统研究不同粒度猫尾草对羔羊体外发酵参数与微生物数量的影响,选取6种不同粒度(1.00、2.36、3.35、4.75、8.00和12.50 mm)的猫尾草作为试验饲粮的纤维来源,按照猫尾草的粒度将试验分为6个处理组,每个处理4个重复,利用Rusitec-s型人工瘤胃模拟装置进行48 h体外发酵,测定养分降解率、发酵参数及发酵液中6种微生物菌群的拷贝数。结果表明：与其他处理组相比,2.36 mm组显著提高了体外中性洗涤纤维降解率(IVNDFD)、丁酸、异丁酸、戊酸、异戊酸摩尔比和溶纤维丁酸弧菌(B. f)数量(P<0.05),且显著降低总产甲烷菌(TMe)数量(P<0.05)。总菌(TB)与氨态氮(NH3-N)浓度、乙酸摩尔比呈正相关(R²=0.84,P=0.038;R²=0.88,P=0.021);B. f数量与丙酸摩尔比呈正相关(R²=0.82,P=0.045),且与pH呈负相关(R²=-0.98,P=0.001);TMe数量与体外粗蛋白质降解率(IVCP...     

噬菌体裂解酶Lysep3的毕赤酵母重组表达和活性分析

试验以突破噬菌体裂解酶Lysep3应用瓶颈为切入点,采用酵母表达系统对其进行重组表达以期降低生产成本,同时初步探究其体外抗菌活性和安全性,以期为临床试验提供数据参考。根据毕赤酵母密码子偏爱性优化并合成Lysep3基因,将其连接至表达载体pPICZαA。经菌落PCR和测序验证后,将线性化重组载体pPICZαA-Lysep3电转化巴斯德毕赤酵母Pichia pastoris X-33。重组酵母转化子经菌落PCR和摇瓶水平甲醇诱导重组蛋白初筛后,在5 L发酵罐中高密度发酵,发酵上清液经His-trap HPGE纯化柱纯化。通过抑菌试验和浊度试验进行rLysep3的抑菌活性分析,通过测定rLysep3对巨噬细胞RAW264.7细胞毒性和小鼠血红细胞溶血性进行rLysep3的安全性分析。结果显示,重组菌在5 L发酵罐中甲醇诱导120 h后Lysep3的表达量达749.2 mg/L。抑菌活性分析发现,rLysep3对肠炎沙门氏菌、金黄色葡萄球菌、猪葡萄球菌及无乳链球菌均具有抑菌活性,且rLysep3可与EDTA联合作用使肠炎沙门氏菌菌落数下降1.6~1.7个数量级。此外,细胞毒性试验发现,rLysep3在1 024 μg/mL浓度下细胞存活率仍可达78.75%;溶血性分析显示,1 024 μg/mL浓度下溶血率仅为1.58%。上述结果证明,通过毕赤酵母表达系统实现了Lysep3的重组表达,rLysep3具有体外抑菌活性,且具低细胞毒性和低溶血性。从抗菌活性、产业化制备及安全性等方面初步显示出rLysep3具有开发为临床治疗禽肠炎沙门氏菌感染的新型抗菌药物的潜力。     

生防木霉菌T2菌株对禾草腐霉病抑菌作用及机制研究

深绿木霉对引起禾草腐霉病原菌瓜果腐霉有明显的抑制作用。深绿木霉菌与瓜果腐霉对峙培养结果表明,深绿木霉菌可以包围、覆盖瓜果腐霉的菌落,同时产生大量孢子;当孢子悬浮液的浓度为4.65×10⁶个/mL时,对瓜果腐霉菌的抑制率达53.56%。通过深绿木霉菌孢子悬浮液和3种药剂对草坪禾草腐霉病病原菌抑制效果的比较,深绿木霉孢子悬浮液浓度为1×10⁷个/mL时与25%甲霜·霜霉的抑菌效果接近,分别为77.37%和79.57%。显微观察结果表明,深绿木霉菌丝缠绕在瓜果腐霉菌菌丝上,穿透菌丝在其内生长;或与瓜果腐霉菌的菌丝平行生长,再侵入瓜果腐霉菌内寄生。深绿木霉菌对瓜果腐霉的主要作用机制为重寄生作用。