生物素对断奶仔猪生产性能及免疫功能影响的研究
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摘要
本研究探讨生物素对PCV2攻击下的断奶仔猪生产性能和免疫功能的影响及其可能机制;探明在PCV2攻击下,断奶仔猪玉米豆粕型饲粮中生物素的适宜添加水平;为建立维生素的抗病营养原理,提供生物素部分的有关理论依据。研究共包括两个动物试验。
试验一饲粮中添加生物素对PCV2攻击下断奶仔猪免疫应答的影响
采用2×3因子试验设计。将48头PCV2抗体阴性、28天断奶、体重为6.8±0.804kg的杜×长×大三元杂交仔猪,随机分成6个处理。每个处理8个重复(公母各半),每个重复1头猪。6个处理分别饲养于2间环控试验舍,即每间3个处理,分别饲喂3种不同生物素水平的饲粮——在玉米豆粕型基础饲粮中,生物素添加水平分别为:0.00、0.05和0.20 mg/kg。一间试验舍的所有试猪在试验第一天经口鼻滴注接种PCV2,另一间用无菌生理盐水假接种。接种后第3、5、7、14、21、28、35天,测定外周血部分免疫指标;接种后第7天起,每周屠宰一头增重率最小的个体,采集肝脏、脾脏、胸腺、腹股沟淋巴组织样,除一份样品制作病理组织切片外,其余在处理后液氮速冻,-80℃保存备用。饲养试验为期35天。结果显示:(1)PCV2攻击刺激了断奶仔猪的细胞免疫应答,生物素0.2mg/kg组的应答效应较强。(2)PCV2攻毒后,PCV2抗体效价高峰阶段大多出现在接种后28-35天,其中生物素0.2mg/kg组在接种后21天进入高效价阶段,并可持续至35天。(3)PCV2攻击降低断奶仔猪血清免疫球蛋白浓度,延迟免疫球蛋白的应答浓度峰值。添加生物素可以增强PCV2攻击下IgM的初次应答强度和时间,提高断奶仔猪血清免疫球蛋白浓度。(4)PCV2攻击显著降低断奶血清IFN-γ浓度,添加生物素可以不同程度地提高血清IFN-γ的浓度,以生物素0.20mg/kg组对IFN-γ提高效果最好。(5)PCV2攻击的全部断奶仔猪淋巴器官组织发生轻到重度病理变化;添加生物素0.20mg/kg对缓减PCV2病毒对淋巴器官组织造成的病理损害效果较好。(6)PCV2攻击下,断奶仔猪第4周和第5周日增重受到显著影响,添加生物素0.20mg/kg组的日增重高于无PCV2攻击处理的0.00mg/kg添加组,添加0.05和0.20mg/kg饲粮生物素分别提高未经PCV2攻击仔猪试验第4周和第5周的日增重。
试验二PCV2攻击下断奶仔猪饲粮中生物素适宜添加水平的研究
采用单因子试验设计:54头PCV2抗体阴性、28d断奶、体重为(6.93±0.721)kg的杜×长×大三元杂交仔猪,随机分成6个处理,每个处理9个重复,每个重复1头猪,试验第一天全部试猪经口鼻接种PCV2病毒。基础饲粮为玉米豆粕型饲粮,在基础饲粮中分别添加生物素0、0.10、0.20、0.30、0.40、和0.50 mg/kg,共6种饲粮处理。试验结果显示:(1)PCV2攻击下,断奶35天,以生物素0.30mg/kg组的E总花环形成率、T淋巴细胞转化率、免疫球蛋白G水平和γ-干扰素水平最高;(2)PCV2攻击下,生物素0.50mg/kg组对生产性能提高的效果最好。(4)PCV2攻击下,添加生物素可以促进外周淋巴器官维持正常形态,提高胸腺生长指数;(5)PCV2攻击下,以添加生物素0.3mg/kg促进IL-2和IFN-γ基因在脾和腹股沟淋巴的表达效果最好。各处理仔猪肝脏MCC基因表达无显著差异,但从趋势上可以看出以0.30mg/kg饲粮添加组提高MCC表达量效果较好。
综上所述,PCV2攻击使断奶仔猪淋巴组织均有不同程度的病理损伤;细胞免疫反应强度减弱,免疫球蛋白水平降低,达到峰浓度的时间被延迟;细胞因子水平下降。添加生物素可以提高细胞免疫强度、免疫球蛋白及细胞因子水平,有利于修复PCV2损伤的淋巴组织。生物素通过在基因水平影响IL-2和IFN-γ,进而影响它们在各免疫器官中的翻译和合成,并通过IL-2和IFN-γ基因表达的合成产物来实现对动物机体的免疫调节。
无PCV2攻击情况下,断奶仔猪玉米豆粕型饲粮中添加生物素0.20mg/kg即可维持一般免疫和生长并具有提高仔猪饲料转化率、降低料肉比的趋势。但在PCV2攻击下或免疫抑制下,仔猪对生物素的需要增加。玉米豆粕饲粮中添加0.30mg/kg生物素提高仔猪的部分免疫指标效果较好,有利于促进IL-2和IFN-γ基因在脾脏和腹股沟淋巴的翻译和合成,从而增强仔猪免疫调节作用的发挥。而0.50mg/kg添加处理组提高生产性能的效果较好。
Two experiments were conducted to study the effects of biotin on immune response in pigs inoculated intranasally with type-2 porcine circovirus,and cytokine IFN-γand IL-2 mRNA gene expression was examined to discuss the mechanism of biotin affecting Weanling pigs' immune function.
Experiment 1:Biotin Supplementation Affects response of immune system of weaned pig of inoculating type-2 porcine circovirus(PCV2)
48 Weanling Duroc×Landrace×Yorkshire crossbred pigs initial BW(6.8±0.804) kg with negative PCV2 antibody were used in a 5-wk trial.Exp.1 was conducted to study the effect of supplemental biotin(0,0.05,and 0.20 mg/kg) and PCV2(chanlenged and un-chanlenged) on IL-2,slL-2R,IFN-γconcentrations,and immune response (humoral and cell-mediated).Although the magnitude of the immune response was small and inconsistent,diets containing 0.05 and 0.20 mg/kg biotin seemed to increase the serum immune globulin levels and the percentage of peripheral blood ANAE~+ T cell.IFN-γconcentration was depressed(P＜0.01) aider PCV2 was challenged,especially on 14 and 21 days postinoculation(dpi),and serum IFN-γlevels were increased(P＜0.02 and P＜0.03) for groups fed 0.05 and 0.20 mg/kg supplemental biotin,respectively,and an interaction(P＜0.01) on the 7 dpi was detected,the IFN-γconcentration were highest for PCV2 chanlenged in combination with the 0.20 mg/kg biotin dietary levels.There was no magnificent effect of biotin level on IL-2 and slL-2R concentrations.PCV2 inoculation depressed the immune response and addition of biotin to the diet tended to increase the immune response.Feeding 0.20mg/kg biotin increased growth performance during the 5-wk trials;PCV2 inoculation depressed growth performance after wk 3.The average daily gain of challenged group of supplemental biotin 0.20 mg/kg was higher than that of the un-challenged group whose supplemental biotin was 0.00 mg/kg.Obvious pathological changes(especially in lung,spleen,stomachs and lymph on mesentery) were observed in all challenged pigs.Biotin added in the basal diet at 0.20mg/kg had an ability of relief of pathological lesion of lymphatic organ caused by virus inoculation.
Experiment 2:Study of the Appropriate Supplementation Level of Biotin in Corn-Soybean Meal diet for Weaned Piglets Challenged With PCV2
The purpose of the Exp 2 was to investigate the appropriate supplementation level of biotin in corn-soybean meal diet for weaned piglets challenged with PCV2.The basal diet was corn-soybean meal diet.There were 6 diets in which boitin was added in the basal diet at 0 mg/kg,0.10 mg/kg,0.20 mg/kg,0.30mg/kg,0.40mg/kg,and 0.50 mg/kg.The results showed as follows:(1) Addition of biotin enhanced significantly T-lymphocyte transformation ratio in blood(P＜0.05),Et-rosette rate and T-lymphocyte transformation ratio of 0.30mg/kg treatment was the greatest on 35 days post-inoculation;(2) The IgG and IFN-γconcentration of 0.30mg/kg treatment were higher significantly on 35 days postinoculation;(3) The growth performance of pigs of 0.50 mg/kg treatment was the best, and the 0.30mg/kg treatment could significantly increased some immuno indexes.(4) With the quantity of supplementation of biotin increasing,the growth indexes of spleen and inguinal lymph nodes of pigs with PCV2 inoculation decreased,on the contrary,the growth indexe of thymus became greater.(5) The treatment of 0.30mg/kg of biotin supplementation level enhanced IL-2 andγ-IFN gene expression,which elevated significantly mRNA expression level of IL-2 andγ-IFN gene(P＜0.01),and there was no significant difference of MCC gene expression in livers of pigs among the biotin addition treatments.
In conclusion,the results suggest that PCV2 challenge results in an immune system response,while biotin supplementation increased,the immune response of weanling piglets. Some cytokines' mRNA expression such as FN-γand IL-2 level changing might be the molecular base of biotin on immune function of piglets.
试验一饲粮中添加生物素对PCV2攻击下断奶仔猪免疫应答的影响
采用2×3因子试验设计。将48头PCV2抗体阴性、28天断奶、体重为6.8±0.804kg的杜×长×大三元杂交仔猪,随机分成6个处理。每个处理8个重复(公母各半),每个重复1头猪。6个处理分别饲养于2间环控试验舍,即每间3个处理,分别饲喂3种不同生物素水平的饲粮——在玉米豆粕型基础饲粮中,生物素添加水平分别为:0.00、0.05和0.20 mg/kg。一间试验舍的所有试猪在试验第一天经口鼻滴注接种PCV2,另一间用无菌生理盐水假接种。接种后第3、5、7、14、21、28、35天,测定外周血部分免疫指标;接种后第7天起,每周屠宰一头增重率最小的个体,采集肝脏、脾脏、胸腺、腹股沟淋巴组织样,除一份样品制作病理组织切片外,其余在处理后液氮速冻,-80℃保存备用。饲养试验为期35天。结果显示:(1)PCV2攻击刺激了断奶仔猪的细胞免疫应答,生物素0.2mg/kg组的应答效应较强。(2)PCV2攻毒后,PCV2抗体效价高峰阶段大多出现在接种后28-35天,其中生物素0.2mg/kg组在接种后21天进入高效价阶段,并可持续至35天。(3)PCV2攻击降低断奶仔猪血清免疫球蛋白浓度,延迟免疫球蛋白的应答浓度峰值。添加生物素可以增强PCV2攻击下IgM的初次应答强度和时间,提高断奶仔猪血清免疫球蛋白浓度。(4)PCV2攻击显著降低断奶血清IFN-γ浓度,添加生物素可以不同程度地提高血清IFN-γ的浓度,以生物素0.20mg/kg组对IFN-γ提高效果最好。(5)PCV2攻击的全部断奶仔猪淋巴器官组织发生轻到重度病理变化;添加生物素0.20mg/kg对缓减PCV2病毒对淋巴器官组织造成的病理损害效果较好。(6)PCV2攻击下,断奶仔猪第4周和第5周日增重受到显著影响,添加生物素0.20mg/kg组的日增重高于无PCV2攻击处理的0.00mg/kg添加组,添加0.05和0.20mg/kg饲粮生物素分别提高未经PCV2攻击仔猪试验第4周和第5周的日增重。
试验二PCV2攻击下断奶仔猪饲粮中生物素适宜添加水平的研究
采用单因子试验设计:54头PCV2抗体阴性、28d断奶、体重为(6.93±0.721)kg的杜×长×大三元杂交仔猪,随机分成6个处理,每个处理9个重复,每个重复1头猪,试验第一天全部试猪经口鼻接种PCV2病毒。基础饲粮为玉米豆粕型饲粮,在基础饲粮中分别添加生物素0、0.10、0.20、0.30、0.40、和0.50 mg/kg,共6种饲粮处理。试验结果显示:(1)PCV2攻击下,断奶35天,以生物素0.30mg/kg组的E总花环形成率、T淋巴细胞转化率、免疫球蛋白G水平和γ-干扰素水平最高;(2)PCV2攻击下,生物素0.50mg/kg组对生产性能提高的效果最好。(4)PCV2攻击下,添加生物素可以促进外周淋巴器官维持正常形态,提高胸腺生长指数;(5)PCV2攻击下,以添加生物素0.3mg/kg促进IL-2和IFN-γ基因在脾和腹股沟淋巴的表达效果最好。各处理仔猪肝脏MCC基因表达无显著差异,但从趋势上可以看出以0.30mg/kg饲粮添加组提高MCC表达量效果较好。
综上所述,PCV2攻击使断奶仔猪淋巴组织均有不同程度的病理损伤;细胞免疫反应强度减弱,免疫球蛋白水平降低,达到峰浓度的时间被延迟;细胞因子水平下降。添加生物素可以提高细胞免疫强度、免疫球蛋白及细胞因子水平,有利于修复PCV2损伤的淋巴组织。生物素通过在基因水平影响IL-2和IFN-γ,进而影响它们在各免疫器官中的翻译和合成,并通过IL-2和IFN-γ基因表达的合成产物来实现对动物机体的免疫调节。
无PCV2攻击情况下,断奶仔猪玉米豆粕型饲粮中添加生物素0.20mg/kg即可维持一般免疫和生长并具有提高仔猪饲料转化率、降低料肉比的趋势。但在PCV2攻击下或免疫抑制下,仔猪对生物素的需要增加。玉米豆粕饲粮中添加0.30mg/kg生物素提高仔猪的部分免疫指标效果较好,有利于促进IL-2和IFN-γ基因在脾脏和腹股沟淋巴的翻译和合成,从而增强仔猪免疫调节作用的发挥。而0.50mg/kg添加处理组提高生产性能的效果较好。
Two experiments were conducted to study the effects of biotin on immune response in pigs inoculated intranasally with type-2 porcine circovirus,and cytokine IFN-γand IL-2 mRNA gene expression was examined to discuss the mechanism of biotin affecting Weanling pigs' immune function.
Experiment 1:Biotin Supplementation Affects response of immune system of weaned pig of inoculating type-2 porcine circovirus(PCV2)
48 Weanling Duroc×Landrace×Yorkshire crossbred pigs initial BW(6.8±0.804) kg with negative PCV2 antibody were used in a 5-wk trial.Exp.1 was conducted to study the effect of supplemental biotin(0,0.05,and 0.20 mg/kg) and PCV2(chanlenged and un-chanlenged) on IL-2,slL-2R,IFN-γconcentrations,and immune response (humoral and cell-mediated).Although the magnitude of the immune response was small and inconsistent,diets containing 0.05 and 0.20 mg/kg biotin seemed to increase the serum immune globulin levels and the percentage of peripheral blood ANAE~+ T cell.IFN-γconcentration was depressed(P＜0.01) aider PCV2 was challenged,especially on 14 and 21 days postinoculation(dpi),and serum IFN-γlevels were increased(P＜0.02 and P＜0.03) for groups fed 0.05 and 0.20 mg/kg supplemental biotin,respectively,and an interaction(P＜0.01) on the 7 dpi was detected,the IFN-γconcentration were highest for PCV2 chanlenged in combination with the 0.20 mg/kg biotin dietary levels.There was no magnificent effect of biotin level on IL-2 and slL-2R concentrations.PCV2 inoculation depressed the immune response and addition of biotin to the diet tended to increase the immune response.Feeding 0.20mg/kg biotin increased growth performance during the 5-wk trials;PCV2 inoculation depressed growth performance after wk 3.The average daily gain of challenged group of supplemental biotin 0.20 mg/kg was higher than that of the un-challenged group whose supplemental biotin was 0.00 mg/kg.Obvious pathological changes(especially in lung,spleen,stomachs and lymph on mesentery) were observed in all challenged pigs.Biotin added in the basal diet at 0.20mg/kg had an ability of relief of pathological lesion of lymphatic organ caused by virus inoculation.
Experiment 2:Study of the Appropriate Supplementation Level of Biotin in Corn-Soybean Meal diet for Weaned Piglets Challenged With PCV2
The purpose of the Exp 2 was to investigate the appropriate supplementation level of biotin in corn-soybean meal diet for weaned piglets challenged with PCV2.The basal diet was corn-soybean meal diet.There were 6 diets in which boitin was added in the basal diet at 0 mg/kg,0.10 mg/kg,0.20 mg/kg,0.30mg/kg,0.40mg/kg,and 0.50 mg/kg.The results showed as follows:(1) Addition of biotin enhanced significantly T-lymphocyte transformation ratio in blood(P＜0.05),Et-rosette rate and T-lymphocyte transformation ratio of 0.30mg/kg treatment was the greatest on 35 days post-inoculation;(2) The IgG and IFN-γconcentration of 0.30mg/kg treatment were higher significantly on 35 days postinoculation;(3) The growth performance of pigs of 0.50 mg/kg treatment was the best, and the 0.30mg/kg treatment could significantly increased some immuno indexes.(4) With the quantity of supplementation of biotin increasing,the growth indexes of spleen and inguinal lymph nodes of pigs with PCV2 inoculation decreased,on the contrary,the growth indexe of thymus became greater.(5) The treatment of 0.30mg/kg of biotin supplementation level enhanced IL-2 andγ-IFN gene expression,which elevated significantly mRNA expression level of IL-2 andγ-IFN gene(P＜0.01),and there was no significant difference of MCC gene expression in livers of pigs among the biotin addition treatments.
In conclusion,the results suggest that PCV2 challenge results in an immune system response,while biotin supplementation increased,the immune response of weanling piglets. Some cytokines' mRNA expression such as FN-γand IL-2 level changing might be the molecular base of biotin on immune function of piglets.
引文
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