巨型艾美尔球虫感染对雏鸡肠道乳酸杆菌的影响

取200只红羽肉杂雏鸡,饲喂至8日龄,随机分为3个球虫卵囊剂量感染组(0.5×105、1.0×105和2.0×105个/羽),并设对照组;于感染后1、3、5、7 d,各组分别处死雏鸡5只,测定小肠中段乳酸杆菌的定植数量。结果表明:与对照组比较3个试验组雏鸡肠道的乳酸杆菌定植数量均有降低。其中试验Ⅰ组,在感染后1、3 d,肠道乳酸杆菌定植数对数值分别为(5.46±0.14)和(5.57±0.14),与对照组比差异不显著(P>0.05);感染5、7 d后,分别为(5.71±0.45)和(5.80±0.52),达到差异显著水平(P<0.05)。试验Ⅱ、Ⅲ组,在感染后1 d,达到差异显著水平(P<0.05)。感染后3、5、7 d,肠道乳酸杆菌定植数量对数值与对照组比较达到极显著水平(P<0.01)。试验证明大剂量球虫感染可明显降低雏鸡小肠段乳酸杆菌的定植数量。     

温度对鸡早熟艾美球虫孢子囊形成的影响

从混合鸡球虫卵囊中分离单卵囊,感染增殖后收获大量单一种类球虫卵囊,将卵囊分别置于10～36℃、38～39℃、40℃恒温箱内培养,研究了温度对鸡早熟艾美球虫孢子囊形成的影响.结果显示,10～36℃时,孢子囊形成时间随温度提高而提前,每小时孢子囊形成率和增长幅度也随温度的提高而上升.10℃时,77 h初次形成孢子囊,孢子囊形成率达19.047%,经98 h孢子囊形成率达92.360%;36℃时,5 h初次形成孢子囊,孢子囊形成率达26.471%,经8 h孢子囊形成率达94.180%;38～39℃时,孢子囊形成率达一定时就不再上升,且随温度提高孢子囊形成率反而下降.38℃时,最高孢子囊形成率为52.800%,39℃时最高孢子囊形成率为12.500%;40℃时,球虫卵囊不会形成孢子囊,39～40℃是球虫卵囊形成孢子囊的临界温度.将卵囊分别置于40～65℃恒温干燥箱内经0.5 h处理后又经25～30℃培养24 h的结果表明,卵囊经40～50℃处理后仍可形成孢子囊,而经55℃以上温度处理就被杀死,45～50℃是球虫卵囊的生死临界温度.     

Pathogenicity of Eimeria praecox alone or associated with Eimeria acervulina in experimentally infected broiler chickens

Eimeria praecox and Eimeria acervulina are two species of coccidia parasites infecting chickens, which develop in the duodenum. The purpose of this study was to evaluate the pathogenicity of E. praecox and to study interactions of this coccidium with E. acervulina. The results showed that the pathogenicity of E. praecox was related to the infective dose, and that its impact on individual weight and weight gain was significant from the lowest administered dose: 5000 oocysts per bird. No morbidity was observed, even with the highest infective dose, but faecal consistency alteration was higher with increasing infective doses. No consistent lesion was observed. When E. praecox was associated with E. acervulina with a low infective dose, performance deterioration seemed to be an additional effect of the two species. However, in the case of heavy infections, signs worsened along with duration of negative impact on growth, compared to a mono-infection.     

In vivo effects of CpG oligodeoxynucleotide on Eimeria infection in chickens

Poultry coccidiosis is the major parasitic disease of poultry and, until now, no recombinant vaccine has been developed. Short oligodeoxynucleotides (ODNs) containing unmethylated CpG motifs (CpG ODNs) have been shown to be effective immunoprotective agents and vaccine adjuvants in mammalian systems. Their use in poultry to protect against intracellular parasites has not been reported to date. The present work investigated the effects of CpG ODN treatment on host susceptibility to Eimeria infection in two chicken strains with different genetic background, SC and TK. The data show that CpG ODN enhanced the birds' resistance to coccidiosis in a normally susceptible chicken strain (TK), as shown by reduced oocyst shedding and improved weight gain. CpG treatment had a differential effect on body weight gains and serum antibody responses, depending on the chicken strain and ODN dose, delivery route, and backbone. This study shows for the first time that CpG ODNs could be used as immunoprotective agents in Eimeria-infected chickens to enhance resistance to the pathogen and improve performance. Future research is needed to optimize their use alone and as vaccine adjuvants that may lead to better and more efficient vaccine applications.     

Column chromatographic characterization of cytoplasmic proteins in Eimeria maxima oocysts from chickens

Cytoplasmic proteins from unsporulated and sporulated Eimeria maxima oocysts were analyzed by gel-filtration column chromatography. Unsporulated oocysts were characterized as having 3 major cytoplasmic proteins and sporulated oocysts as having 5 major cytoplasmic proteins. Molecular weights ranged from 5 x 10(3) to 1.4 x 10(6). Larger molecular weight proteins were detected in sporulated and unsporulated oocysts, but were associated more with sporocysts of sporulated oocysts.     

Acquisition of immunity to Eimeria maxima in newly hatched chickens given 100 oocysts

The acquisition of immunity to Eimeria maxima by chicks infected 18 hr after hatch with a single dose of 100 oocysts was investigated. In the first experiment, birds were moved each day to clean cages in order to prevent the possibility of secondary infection resulting from ingestion of oocysts passed in their feces. Immunity was measured at 4 wk of age by calculation of oocyst production following challenge with 500 oocysts or weight gain following challenge with 100,000 oocysts. Large numbers of oocysts were produced by infected birds following challenge, although numbers were significantly less than those from birds that had been reared in the absence of infection (susceptible controls). The weight gain of infected birds following challenge was significantly greater than that of susceptible controls but less than that of unchallenged controls. Thus, only partial protection had been acquired, whether parasite replication or body weight gain was used to assess the extent of immunity development. In a second experiment, acquisition of immunity at 4 wk by chicks infected 18 hr after hatch with 100 oocysts of E. maxima and reared in floor pens in contact with their droppings was investigated. Infected birds produced no oocysts following challenge, and weight gains were not significantly different from the unchallenged controls, which indicates that full immunity had developed by 4 wk. It is concluded that if oocysts of Eimeria species are used to vaccinate day-old chicks, reinfection by oocysts present in the litter is necessary for the establishment of protective immunity.     

Endocrine changes in the fowl during infection with Eimeria maxima

Plasma concentrations of thyroxine (T4), triiodothyronine (T3), prolactin, growth hormone and corticosterone were measured in immature domestic fowl which were infected with Eimeria maxima, uninfected but given the same amount of food as those infected (pair-fed) or fed ad libitum. Infection with E maxima caused a decrease in plasma T4 concentration whereas pair feeding caused an increase when food intake was minimal. Plasma T3 concentrations were decreased similarly in both infected and pair-fed birds. Infection caused an increase in plasma prolactin concentration, whereas pair-feeding did not. The plasma concentration of growth hormone was not affected by any of the treatments. Plasma corticosterone concentration was markedly increased on days 5 and 6 in pair-fed birds but was not affected by infection with E maxima.     

Guar meal ameliorates Eimeria tenella infection in broiler chicks

Guar meal contains relatively high levels of saponins, which are known to have antiprotozoal activity and may be effective against coccidiosis. A 2x2 factorial experiment investigated the impact of guar meal (0 or 5%) corn-soy-based starter broiler diets on chicks unchallenged or challenged with Eimeria tenella. At 1 day of age, 120 unsexed RossxRoss broiler chicks were randomly distributed among four treatment groups. Chicks were challenged with 5x10(3) sporulated oocysts of E. tenella in 0.5ml at 10 days of age by oral gavage. Weekly body weight, body weight gains, feed conversion ratio and mortality rate were recorded for chicks fed from 0 to 21 days of age. Oocysts shed per gram feces were recorded from 6 to 10 days post-challenge. Results showed that challenged chicks fed 0% guar meal had significantly higher oocysts per gram shed in feces than the other groups. No significant differences among treatment groups in mortality rate were observed. Body weights of unchallenged and challenged chicks fed 0% guar meal were significantly higher than those fed 5% guar meal at 2 weeks of age. Results indicated that including 5% guar meal in the diet of chicks challenged with E. tenella decreased oocysts shed per gram feces and prevented bloody diarrhea, but without affects on body weight and feed conversion ratio at 11 days post-challenge.     

中药复方制剂预防鸡巨型艾美耳球虫感染的效果观察

为了探讨常山、青蒿、黄连、大黄、黄芩、黄柏、黄芪、地榆、柴胡等中药组成的不同中药复方制剂预防鸡巨型艾美耳球虫感染的效果,试验设阴性对照组(Ⅰ组)、阳性对照组(Ⅱ组)、中药预防Ⅲ组、中药预防Ⅳ组、中药预防Ⅴ组和中药预防Ⅵ组6个组。结果显示,6个组相对增重率依次为100%、46.65%、84.94%、46.81%、11.37%和30.77%,攻虫后各组死亡率依次为0、20%、10%、0、10%和0,肠道病变记分依次为0、2.4、1、1、1.2和0.8;抗球虫指数(ACI)6个组依次为200、80.25、161.94、121.81、86.67和97.17。结果表明,中药复方制剂Ⅰ预防球虫病效果较好,其他组也有一定的预防作用,但效果不明显。该试验结果为中药复方制剂预防鸡球虫病提供理论依据。     

Protective effects of sugar cane extracts (SCE) on Eimeria tenella infection in chickens

The effects of oral administration of sugar cane extracts (SCE) on Eimeria tenella oocysts infection in chickens were studied with 2 different experiments. In Experiment 1, 3-week-old inbred chickens (MHC; H.B15) were inoculated into the crop with SCE (500 mg/kg/day) for 1 day or 3 consecutive days, and then challenged with E. tenella sporulated oocysts (2 x 10(4) cells/chicken). In Experiment 2, 1-week-old chickens were orally administered SCE at the same dose for 3 consecutive days, and then initially infected with E. tenella sporulated oocysts (2 x 10(3) cells/chicken). At 2 and 3 weeks of age, these chickens were immunized intravenously with the mixed antigens of sheep red blood cells (SRBC) and Brucella abortus (BA). At 4 weeks of age, chickens were challenged with E. tenella sporulated oocysts (1 x 10(5)/chicken). Challenged chickens with E. tenella oocysts showed markedly decreased body weight gain/day, severe hemorrhage and great number of shedding oocysts in feces and high lesion scores. Oral administration of SCE and initial infection with oocysts (2 x 10 (3)/chicken) resulted in a remarkable improvement in body weight gain/day, hemorrhage, the number of shedding oocysts and lesion score, compare to other infected groups. In addition, SCE-inoculated chickens with the initial infection showed a significant increase in antibody responses against SRBC and BA and also improvement in decreased relative proportions of Bu-1a(+) and CD4( )cells in cecal tonsil lymphocytes of E. tenella-challenged chickens. Cecal tissues of chickens administered SCE and initially infected with E. tenella oocysts showed lower numbers of schizonts, gametocytes and oocysts than those of infected control chickens. These results suggest that SCE have immunostimulating and protective effects against E. tenella infection in chickens.     

Factors influencing enhanced Salmonella typhimurium infection in Eimeria tenella-infected chickens

To investigate a possible mechanism involved in the enhancement of Salmonella typhimurium infection in chickens concurrently infected with Eimeria tenella, S typhimurium was given orally to chickens 7 days after E tenella inoculation. The number of viable S typhimurium decreased in the ceca of chickens not inoculated with E tenella, whereas the number gradually increased in the ceca of chickens inoculated with E tenella. Cecal contents were analyzed for pH value, oxidation-reduction potential, and amounts of short-chain fatty acids and bile acids. In the ceca of E tenella-inoculated chickens, the oxidation-reduction potential significantly (P less than 0.05) shifted to the oxidative phase, and the concentration of volatile fatty acids (acetic acid, propionic acid, and butyric acid) significantly (P less than 0.05) decreased. In both aerobic and anaerobic incubations, the number of viable S typhimurium in vitro decreased as the molar concentration of fatty acids increased. Experimental evidence indicated that multiplication of S typhimurium in the ceca of E tenella-inoculated chickens was associated with decreased concentrations of volatile fatty acids.     

Effects of a herbal complex against Eimeria tenella infection in chickens

A liquid and a powder made from a herbal complex consisting of Uncariae Ramulus cum Uncis, Agrimoniae Herba, Sanguisorbae Radix, Eclipta Prostrate Herba, Pulsatillae Radix, Sophorae Flavescentis Radix, Rehmanniae Radix and Glycyrrhizae Radix were studied for their anticoccidial activities in chickens. Chickens were administered with herbal liquid, powder, diclazuril or without medication during the study and challenged with oocysts of Eimeria tenella. Results indicated that the birds medicated showed less bloody faeces than those without medication. The intestinal lesion was mild in the chicks medicated with herbal liquid without significantly different lesion score when compared with uninfected chicks. The birds with medication had significantly higher body weight gains than birds without medication. Therefore, the herbal complex used in this study was effective against E. tenella infection in chickens.     

Effects of Natustat supplementation on performance, feed efficiency and intestinal lesion scores in broiler chickens challenged with Eimeria acervulina, Eimeria maxima and Eimeria tenella

The effects of dietary supplementation of Natustat™, a propriety plant derived product (Alltech Inc., KY, USA) and Salinomycin, on performance, feed efficiency and intestinal lesion scores were observed during two Eimeria challenge trials in broiler chickens. In the first trial chickens were challenged with Eimeria sp. via infecting the litter with a known amount of Eimeria oocysts. In the second trial the source of the Eimeria challenge was the litter from the first trial and the same treatment groups were assigned to the same pens as in the initial trial.

Birds were placed 55 per pen with seven pens per treatment. Performance parameters were recorded on days 21 and 42 during both trials. Intestinal lesion scores were assessed on days 14 and 21 during Trial 1 and on day 21 during Trial 2. Average weight gain and feed conversion ratios were significantly improved in the Natustat™ and Salinomycin treatment groups when compared to the non-supplemented infected group. Furthermore, lesion scores were lower on all sampling days in the Natustat™ and Salinomycin groups when compared to the non-supplemented group. However, only lesions associated with Eimeria tenella were significantly lowered by Natustat™ and Salinomycin supplementation.

Natustat™ and Salinomycin were equivalent in alleviating the negative performance effects associated with coccidiosis challenge. In summary, Natustat™ has the potential to be used as a natural alternative to chemotherapeutic drugs for Eimeria control.     

巨型艾美耳球虫抗原Em8对鸡外周血单核细胞功能的影响

探讨了巨型艾美耳球虫抗原Em8体外对鸡外周血单核细胞(PBMCs)功能的影响。选取抗原基因Em TFP250中具有较好免疫保护性的Em8基因片段,并对其进行原核表达纯化出重组蛋白。无菌采集未被鸡球虫感染的鸡的外周血,分离出单核细胞,加入终浓度为5μg/m L的Em8重组蛋白进行体外培养,通过免疫荧光抗体技术观察重组蛋白与单核细胞的结合情况;Em8重组蛋白不同浓度体外刺激鸡外周血单核细胞,用q PCR技术测定各实验组单核细胞中IL-4、IL-17D、IFN-γ、TGF-β4的mRNA转录水平;采用CCK-8法测定细胞的增殖情况;Em8重组蛋白不同浓度(低、中、高浓度)体外与巨噬细胞共培养,测定巨噬细胞吞噬和NO分泌功能。结果表明:重组蛋白Em8是巨型艾美耳球虫的重要抗原,发挥广泛而重要的免疫作用。其中,重组蛋白Em8能够与外周血单核细胞结合并能刺激细胞因子IL-17D和IFN-γ的表达水平显著提高(P0.01),能够引起单核细胞增值(P0.01),巨噬细胞吞噬作用增强(P0.01)且NO分泌增加(P0.01)。     

Kinetic differences in intestinal and systemic interferon-gamma and antigen-specific antibodies in chickens experimentally infected with Eimeria maxima

Kinetic differences between systemic vs. intestinal and humoral vs. cellular immune responses were elucidated in chickens experimentally infected with Eimeria maxima by comparing interferon-gamma (IFN-gamma) and parasite-specific antibody levels in the intestine and serum during the course of infection. The level of serum IFN-gamma correlated significantly with fecal oocyst shedding (r2 = 0.97), thereby establishing the importance of cell-mediated immunity in coccidia infection. Moreover, intestinal IFN-gamma levels increased sooner than those in sera (4 vs. 6 days postinfection) and both were observed prior to the appearance of parasite-specific antibodies (8-10 days postinfection), again indicating the importance of intestinal cellular immunity in coccidiosis. Although immunoglobulin (Ig)G, IgA, and IgM isotypes of the antigen-specific antibody response increased significantly in both the intestine and serum after E. maxima infection, intestinal IgA-specific antibodies showed the most dramatic increase. However, the relevance of this observation in the context of primary Eimeria infection is unclear because the coccidia parasites have reached the final stages of their life cycle by this time. These results thus demonstrate the importance of T-cell immune responses against coccidia, characterized by local IFN-gamma secretion in the intestine, in mediating host protective immune response to coccidia.     

巨型艾美耳球虫（Eimeria maxima）早熟弱毒株的选育

近 30年来 ,鸡球虫的免疫预防研究取得了令人鼓舞的进展。国外已有 4种球虫苗进入商品市场 ,国内也在积极开展免疫预防的研究。本教研组已研制出鸡球虫活苗 ,现正进行早熟苗的研制工作 ,已成功选育出 E.tenella,E.acervulina和 E.Brunetti的早熟弱毒株。本研究对致病性较强的 E.maxima进行早熟弱毒株选育 ,并对所选早熟株的生物学特性进行研究。1　材料与方法1 .1　实验动物　刚出壳蛋鸡公雏 ,购自中国农科院畜牧所 ,饲养于无球虫笼舍中 ,饲喂无抗球虫添加剂的无球虫污染的全价饲料 (由中国农科院畜牧所配制 ) ,饮水用自来水 ,试验前 ,检…     

Antioxidant status during the course of Eimeria tenella infection in broiler chickens

The antioxidant status of broiler chickens (Cobb 500 hybrids) infected with Eimeria tenella was monitored by determining blood plasma malondialdehyde (MDA), superoxide dismutase (SOD) and catalase (CAT). The results of the experiment showed an increase in MDA - a marker of radical-induced damage of E. tenella-infected birds, compared to healthy chickens (3.01 micro mol/L vs. 2.55 micro mol/L, P<0.05). Correspondingly, a decreased SOD activity was observed in infected birds compared to controls (2429.0 U/g Hb vs. 3044.6 U/g Hb, P<0.05). Furthermore, CAT activity in infected birds was higher than in healthy ones (2242.2 U/g Hb vs. 1367.0 U/g Hb, P<0.001). The observed enzyme changes suggest an impaired antioxidant status of chickens during the course of an E. tenella infection and the occurrence of oxidative stress following infection. Alterations in the caecum, oocyst production, weight gain and feed conversion ratio were indicative of a severe infection involving pathogenic oxidative stress and impaired ecological oxidative balance.     

Does Plasmodium gallinaceum induce relapse of Eimeria acervulina infection in chickens?

An experiment was carried out to test the hypothesis that chickens previously infected with Eimeria acervulina, but having ceased producing oocysts, recommence E acervulina oocyst production when infected with Plasmodium gallinaceum. No relapse of coccidiosis was caused by the malarial infection. Subsequent treatment with the immunosuppressant betamethasone of the control chicks infected with E acervulina only did not reveal any occult coccidial infection. The results are critically compared with previously published results which apparently supported the hypothesis.