Influence of reticuloendotheliosis on the severity of Eimeria tenella infection in broiler chickens

Broiler chickens free of maternal immunity to reticuloendotheliosis virus (REV) were used in the experiment. Two groups of 25 chickens were inoculated with REV at one day of age. One of these groups and another group of 25 chickens were inoculated with Eimeria tenella sporulated oocysts at 7 days of age. Chickens inoculated with E. tenella showed bloody diarrhoea from 12 to 14 days of age. Six out of 25 chickens died (P less than 0.05) at 13 and 14 days of age in the dual infected group. At 14 days of age, when chickens were killed, the lesion score in the combined infection group, was statistically different from that in the chickens inoculated with E. tenella alone. Also the weights of the bursa of Fabricius and thymus were lower in the two REV infected groups than in the controls. Although REV infection alone adversely affected the weight gain and feed conversion, with combined infection this effect was much greater. Following REV inoculation most of the chickens showed feathering defects and all the examined chickens were viraemic at 21 days of age. At the same age, all but one chicken failed to show precipitating antigenaemia and about one-half of these chickens showed a very low serum neutralisation titre. None of these chickens showed precipitating antibodies.     

Evaluation of interspecific hybrids of the chicken, guinea fowl, and Japanese quail for innate resistance to coccidia

Experimental chicken/guinea fowl hybrids, guinea fowl, and chickens were orally inoculated with Eimeria acervulina or E. tenella, which are specific for chickens, or with E. grenieri, which is specific for guinea fowl. No intact oocysts were found in feces within 24 hr of inoculation, suggesting that excystation occurred in the normal and abnormal hosts. No oocysts were found in the feces of hybrids during a 9-day postinoculation period. The guinea fowl passed oocysts of guinea fowl coccidia (E. grenieri) but not those of chicken coccidia, and the chickens passed oocysts of chicken coccidia (E. acervulina and E. tenella) but not those of guinea fowl coccidia. Some asexual development (schizogony) occurred in hybrids inoculated with E. tenella, but sexual development (gametogony) did not. In contrast, quail/chicken hybrids became infected with oocysts of chicken coccidia (E. acervulina, E. tenella, and E. maxima) and quail coccidia (E. bateri) and passed a few oocysts during the normal patent period; control chickens and quails became heavily infected with oocysts of chicken and quail coccidia, respectively.     

三株柔嫩艾美球虫野外分离株的致病性分析

用每鸡4×104和8×104个孢子化卵囊的剂量感染黄羽肉雏,以临床症状、肉眼病变、平均增重、病变记分和死亡率为判定指标,对柔嫩艾美球虫EtCZ-1、EtCZ-2、EtCZ-3株的致病性进行了比较分析。结果显示:各虫株引起的临床症状和肠道肉眼病变基本相似,致病性随着感染剂量的增加而增强。各感染组的增重与对照组相比均有显著差异(P<0.50),相对增重率分别下降了74.4%～83.2%、68.7%～72.2%和70.6%～74.4%;感染后鸡出现血便的时间较一致,即第108小时开始排血便,在120h～144h达高峰;病变值和卵囊值均随着感染剂量的增加而增大;除EtCZ-1株高剂量组的存活率是90%以外,其余各组均为100%。显示3株球虫的致病性相似。     

藏鸡和隐性白羽鸡感染柔嫩艾美耳球虫后免疫相关基因的表达变化分析

试验旨在从免疫遗传学的角度初步探讨藏鸡（TC）和隐性白羽鸡（RWC）对柔嫩艾美耳球虫（Eimeria tenella）易感性差异的分子机制,分别用1×10⁵个柔嫩艾美耳球虫孢子化卵囊感染对球虫具有抗性的藏鸡和易感的隐性白羽鸡。应用实时荧光定量PCR检测藏鸡和隐性白羽鸡感染前0 d和感染后第2、4、6和8天脾脏、盲肠、胸腺、法氏囊中γ-干扰素（IFN-γ）、白细胞介素-2（IL-2）、IL-16、Toll样受体（TLR3）和TLR15免疫相关基因的转录水平变化。结果显示,藏鸡脾脏IFN-γ、IL-2、IL-16及TLR3、TLR15免疫相关基因转录水平于感染后第4和8天明显上调,隐性白羽鸡则无明显变化。藏鸡盲肠IFN-γ转录水平在感染后第2天显著上调（P<0.05）,TLR3在感染后第4天起显著上调（P<0.05）,其余免疫相关基因变化幅度不大;隐性白羽鸡盲肠IFN-γ转录水平在感染后第8天显著上调（P<0.05）,IL-2在感染后第2天起显著上调（P<0.05）,IL-16在感染后第6天起显著上调（P<0.05）,TLR3在感染后第2和8天显著上调（P<0.05）,TLR15变化幅度不大。各免疫相关基因在2个品种鸡胸腺和法氏囊中均出现上调或下调,但除藏鸡法氏囊TLR3和TLR15转录水平变化幅度相对较大外,其余免疫相关基因与感染前相比变化幅度不大。以上结果显示,球虫感染主要导致藏鸡和隐性白羽鸡脾脏和盲肠中的各免疫相关基因出现显著变化,表明宿主的遗传背景在一定程度上可影响球虫感染的免疫应答。     

外源性一氧化氮对鸡柔嫩艾美尔球虫的作用

用酸化亚硝酸钠作为一氧化氮(NO)供体处理新收取的柔嫩艾美尔球虫未孢子化和已孢子化卵囊,来探讨外源性NO对球虫卵囊的抑杀作用。另外通过内服亚硝基谷胱甘肽(GSNO)试验来观察外源性NO对球虫病的防治作用。结果表明,酸化亚硝酸钠溶液(pH=1)对球虫卵囊的孢子生殖具有明显的抑制作用,20mmol/L酸化亚硝酸钠溶液对卵囊孢子生殖的抑制率可达100%;但不经酸化的亚硝酸钠溶液和相同pH的盐酸溶液对卵囊的孢子生殖没有明显影响。经20mmol/L以上浓度酸化亚硝酸钠处理的孢子化卵囊也失去了对雏鸡的致病力,但直接内服GSNO溶液则没有明显的抗球虫作用。这提示外源性NO对鸡柔嫩艾美尔球虫未孢子化和孢子化卵囊均有抑杀作用,但直接内服没有明显的抗球虫病作用。     

柔嫩艾美耳球虫广西南宁株的分离及鉴定

利用从南宁市郊养鸡场球虫病鸡粪便中收集的球虫混合种卵囊感染小鸡,再应用单卵囊分离感染技术,从感染鸡盲肠中收集的卵囊分离纯化获得1株纯种球虫,经鸡体传代增殖,对该虫株的卵囊大小和卵形指数、潜在期、排卵高峰期、最短孢子化时间、寄生部位、致病性等指标进行观察和测定。结果测得该虫株卵囊的平均大小为(25.743±1.94126)μm×(21.4±1.85985)μm,平均卵型指数为1.2067±0.07;潜在期为140 h;其排卵囊峰期在第6～9天,最高峰在第7天;最短孢子化时间为19 h;寄生部位在盲肠;对两周龄的艾维茵鸡,当使用5×104的孢子化卵囊感染剂量时死亡率为7.5%。根据这些测定和观察到的指标综合鉴定该分离株球虫为柔嫩艾美耳球虫,并命名为柔嫩艾美耳球虫广西南宁株(Eimeria.tenella-GXNN),该研究结果为进一步研究本地区鸡球虫病的药物治疗和免疫预防等奠定了基础。     

An enzyme-linked immunosorbent assay for coccidiosis in chickens: correlation of antibody levels with prior exposure to coccidia in the laboratory and in the field

An enzyme-linked immunosorbent assay was developed for detecting antibody to coccidia to facilitate the survey of laboratory and field infections. Serum antibody levels in chickens were measured against soluble Eimeria tenella oocyst antigen. Sera from breeders aged 10, 23, 37, and 43 weeks were positive with uniformly high antibody titers. Broiler chick sera showed high maternal antibody titer at hatch, decreasing to an almost negligible response at 3 weeks of age. Two-week-old broiler chicks had variable responses to a single infection of E. tenella: titers were elevated at 8 to 10 days postinfection and generally increased through day 24. Weekly reinfection of 2-week-old broiler chickens produced an antibody titer in proportion to the number of oocysts per dose and stimulated protection against challenge with 2 x 10(5) E. tenella. Inbred birds raised in a pathogen-free environment for 6 weeks had no detectable antibody titers.     

Effect of dietary supplementation with oregano essential oil on performance of broilers after experimental infection with Eimeria tenella.

A study was carried out to examine the effect of dietary supplementation of oregano essential oil on performance of broiler chickens experimentally infected with Eimeria tenella at 14 days of age. A total of 120 day-old Cobb-500 chicks separated into 4 equal groups with three replicates each, were used in this study. Two groups, one infected with 5 x 10(4) sporulated oocysts of E. tenella and the other not, were given a basal diet and served as controls. The other two groups also infected with E. tenella were administered diets supplemented with oregano essential oil at a level of 300 mg/kg, or with the anticoccidial lasalocid at 75 mg/kg. Following this infection, survival rate, bloody diarrhoea and oocysts excretion as well as lesion score were determined. Throughout the experimental period of 42 days, body weight gain and feed intake were recorded weekly, and feed conversion ratios were calculated. Two weeks after the infection with E. tenella supplementation with dietary oregano oil resulted in body weight gains and feed conversion ratios not differing from the non-infected group, but higher than those of the infected control group and lower than those of the lasalocid group. These parameters correspond with the extent of bloody diarrhoea, survival rate, lesion score and oocyst numbers and indicated that oregano essential oil exerted an anticoccidial effect against E. tenella, which was, however, lower than that exhibited by lasalocid.     

Isolation and selection of ionophore-tolerant Eimeria precocious lines: E. tenella, E. maxima and E. acervulina

Eimeria parasites were isolated from Nanhai Guangdong province (southern China) and studied in chickens in wire cages to evaluate their drug resistance against commonly used ionophores: monensin (100 mg/kg of feed), lasolacid (90 mg/kg), salinomycin (60 mg/kg), maduramicin (5 mg/kg) and semduramicin (25 mg/kg). Chinese Yellow Broiler Chickens were infected with 40,000 crude sporulated Eimeria oocysts at 15 days of age and prophylactic medication commenced a day prior to infection. Drug resistance was assessed for each ionophore drug by calculating the anticoccidial index (ACI) and percentage optimum anticoccidial activity (POAA) based on relative weight gain, rate of oocyst production and lesion values. Results revealed that Nanhai Eimeria oocysts comprising of E. tenella, E. maxima and E. acervulina, were resistant to monensin, sensitive to both salinomycin and lasolacid and partially sensitive to maduramicin and semduramicin. By selection for early development of oocysts during passage through chickens, the prepatent time of E. tenella, E. maxima and E. acervulina were reduced by 49, 36 and 22 h, respectively. The precocious lines are less pathogenic than the parent strains from which they were selected and conferred a satisfactory protection for chickens against coccidiosis. These ionophore-tolerant precocious lines could have wider applications in the development of anticoccidial vaccines for sustainable control of coccidiosis.     

Anticoccidial effect of green tea-based diets against Eimeria maxima

Anticoccidial effects of green tea (GT)-based diets were evaluated in chickens following oral infection with Eimeria maxima an ubiquitous intestinal parasite of poultry that impairs the growth and feed efficiency of infected birds. Five-week-old chickens were assigned to four groups (GT 0.5%, GT 2.0%, untreated/infected and non-infected control) and each group consisted of 15 chickens. Chickens were fed a standard diet supplemented with ground green tea for 2 weeks prior to infection with E. maxima (10,000 sporulated oocysts per bird). The effects of green tea on E. maxima infection were assessed by two parameters, fecal oocyst shedding and body weight gain. The green tea-fed chickens produced significantly reduced fecal oocysts (P<0.05) when compared to the E. maxima-infected group fed standard diet. The green tea-based diet, however, did not improve body weight loss caused by E. maxima infection. This study is the first to demonstrate anticoccidial effect of green tea on Eimeria parasites.     

Studies on the pathogenesis of chicken infectious anaemia virus infection in six-week-old SPF chickens

The pathogenesis of chicken infectious anaemia virus (CAV) infection was studied in 6-week-old and one-day-old SPF chickens inoculated intramuscularly with graded doses of Cux-1 strain (10(6)-10(2) TCID50/chicken). Viraemia, virus shedding, development of virus neutralizing (VN) antibodies and CAV distribution in the thymus were studied by virus isolation, polymerase chain reaction (PCR), immunocytochemistry (IP) and in situ hybridization until postinfection day (PID) 28. In 6-week-old chickens infected with high doses of CAV, viraemia and VN antibodies could be detected 4 PID and onward without virus shedding or contact transmission to sentinel birds. However, virus shedding and contact transmission were demonstrated in one-day-old infected chickens. In the 6-week-old groups infected with lower doses, VN antibodies developed by PID 14, transient viraemia and virus shedding were detected. The thymus cortex of all 1-day-old inoculated chickens stained with VP3-specific mAb. Cells with positive in situ hybridization signal were fewer and scattered throughout the thymus tissue of the one-day-old inoculated chickens as compared to IP-positive cells. These results suggest that early immune response induced by high doses of CAV in 6-week-old chickens curtails viral replication and prevents virus shedding.     

鸡胚接种柔嫩艾美耳球虫和堆形艾美耳球虫活卵囊的免疫保护试验

给18日龄鸡胚接种一定剂量的柔嫩艾美耳球虫(Eim eria tenella)和/或堆形艾美耳球虫(E.acervulina)孢子化卵囊,出雏后在无球虫环境中笼养,1~10日龄每天收集各组粪便样本,计数克粪便卵囊数(OPG),并于14日龄时以大剂量同源孢子化卵囊攻虫,以相对增重率(RWG)、饲料转化率(FCR)、相对卵囊产量(ROP)评价免疫保护效果。结果显示,以E.tenella或E.acervulina卵囊免疫18日龄鸡胚,其卵囊排出的潜隐期及达到峰值的时间与1日龄雏鸡接种组相一致,有相似的排卵囊曲线,提示其诱导免疫的建立是在出雏后开始建立的。攻虫后各免疫组的RWG由攻虫对照组的31.9%~51.7%提高到了76.5%~83.6%,RCR由攻虫对照组的4.11~4.89改善为2.72~2.96,ROP降至4.7%~23.5%。结果表明以一定剂量E.tenella和E.acervulina卵囊单独或混合经羊膜腔免疫18日龄鸡胚都可以建立起针对出雏后14日龄同源攻虫的良好免疫保护力。比较混合免疫E.tenella和E.acervulina卵囊组与单一接种E.tenella或E.acervulina卵囊组的免疫效果发现,混合免疫组的各项指标均稍优于后者。     

青蒿散抗鸡球虫药效研究

为探讨青蒿散对鸡柔嫩艾美耳球虫（E.tenella）的防治效果,本试验选取90只健康黄羽肉鸡,分为6组,除空白对照组外,其余各组（感染对照组、药物对照组及青蒿散低、中、高剂量组）试验鸡均经口感染6×10⁴个柔嫩艾美耳球虫孢子化卵囊,各试验组在接种前一天开始用药至试验结束。记录各组试验鸡的眼观病变、组织病理学检查、相对增重率、盲肠病变记分、血便记分、每克粪便中的卵囊数（OPG）、卵囊值及抗球虫指数（ACI）指标,评价青蒿散不同给药浓度的抗球虫效果。结果显示,空白对照组、感染对照组、药物对照组及青蒿散低、中、高剂量组的抗球虫指数分别为200.00、72.23、157.19、82.89、125.32和137.06,与感染对照组相比,药物对照组和青蒿散各剂量组的眼观病变、组织病理学检查均有所缓解,青蒿散能减轻病鸡盲肠的肿胀和出血等症状,且病鸡精神状态较感染对照组好;与感染对照组相比,各给药组的平均增重均有所升高,盲肠病变、卵囊值及血便情况减少。综上所述,青蒿散能缓解柔嫩艾美耳球虫侵染鸡盲肠所致的病理症状和组织病变,减少粪便中卵囊的排出数量,有一定的抗球虫效果,且给药浓度越高抗球虫效果越好,具有剂量依赖性。     

雏鸡感染柔嫩艾美耳球虫后脂质过氧化物的变化

为了探讨柔嫩艾美耳球虫 (E.tenella)对感染雏鸡的损伤机制 ,本试验给 10日龄海兰白公雏鸡口服柔嫩艾美耳球虫孢子化卵囊 8× 10 4个 /只 ,在感染后 2、4、6、7、9、12、16和 2 1d分别各取 5只雏鸡剖检 ,取盲肠并采血 ,观察盲肠病理组织学变化 ,测定盲肠组织及血清中的丙二醛 (MDA )、超氧化物歧化酶 (SOD)、谷胱甘肽过氧化物酶 (GSH- Px)的水平。结果表明 ,雏鸡在感染 E.tenella后 2～ 4d,盲肠开始出现损伤 ,同时 SOD和 GSH- Px活性也开始下降 ;感染后 4～ 7d,盲肠粘膜上皮和肠腺上皮细胞大量变性、坏死崩解 ,发生严重损伤时 ,盲肠及血清中 MDA含量显著增加 ,并持续到 9d,同时 SOD和 GSH- Px活性也显著下降 ;感染 9d后 ,损伤的盲肠组织及 MDA、SOD和 GSH- Px水平逐渐恢复 ,至 2 1d基本达到正常     

柔嫩艾美尔球虫感染对鸡的若干生理生化指标的影响

以74只33日龄蛋公鸡作为试验对象,随机分成感染组(n=37)和对照组(n=37)。感染组每只口服接种1.5×10~5个柔嫩艾美尔球虫孢子化卵囊,分别于感染前(0d)和感染后(2,4,6,8d)将鸡处死,颈静脉采血,研究球虫感染对鸡生理生化指标的影响。结果表明柔嫩艾美尔球虫感染后鸡红细胞数、白细胞数、血红蛋白含量、血清中葡萄糖含量显著减少,血清钾含量显著升高,血清钠、血清钙、氯化物、无机磷都显著降低。血清镁含量也降低,但与对照组相比无统计学差异。     

Effect of ibuprofen on coccidiosis in broiler chickens

Ibuprofen (IBU)-a nonsteroidal anti-inflammatory drug-inhibits the biosynthesis of prostaglandins with pro-inflammatory and immunosuppressive properties and is therefore proposed as a candidate molecule for the treatment of coccidiosis in broiler chickens. In all experiments, IBU was administered via drinking water. In a first experiment, chickens were infected at 10 or 21 days of age with oocysts of Eimeria acervulina (5 X 10(4)), Eimeria maxima (3 X 10(4)), and Eimeria tenella (7.5 X 10(3)) and medicated with IBU at a dose of 15 mg/kg body weight (BW). In a second experiment, chickens were infected at 6 days of age with 10(4) oocysts of E. acervulina and medicated with IBU at a dose of 100 mg/kg BW. In the third experiment, an inoculum consisting of 5 x 10(4) or 10(5) E. acervulina oocysts was administered at 6 days of age to chickens medicated with IBU at a dose of 100 mg/kg BW. In a fourth experiment, the effect of IBU on sporulation and infectivity of E. acervulina oocysts was studied. Coccidial lesion scores (CLSs), oocyst shedding, and weight gain were used as evaluation parameters in all experiments except the fourth, where weight gain was not taken into account. In addition, the sporulation percentage was determined in the last experiment. No influence of IBU on the indicated parameters was observed after providing the drug at a dose of 15 mg/kg BW, whereas CLSs and oocyst shedding were reduced when IBU was provided at a dose of 100 mg/kg BW. However, IBU did not significantly show any effect on the degree of sporulation and infectivity of E. acervulina oocysts at a dose of 100 mg/kg BW.     

Isolation, propagation, identification and comparative pathogenicity of five Egyptian field strains of Eimeria tenella from broiler chickens in five different provinces in Egypt

A battery trial was conducted to evaluate the comparative pathogenicity of five field strains of Eimeria tenella from Behera, Khafr El-Sheikh, Alexandria, Gharbia and Matrouh provinces in Egypt. Two-week-old broiler chickens were infected with 25×10(3) sporulated oocysts of each strain of E. tenella. The comparative pathogenicity of the strains was assessed by calculating body weight gain, feed conversion ratio, lesion scores, dropping nature scores, cecal scrapings, mortality percentage and oocysts count. Hematological parameters including hemoglobin (Hb) content, packed cell volume (PCV%) and total erythrocytic count, were also evaluated. There were different degrees of pathogenicity between the strains.     

Assessment of dietary supplementation with probiotics on performance, intestinal morphology and microflora of chickens infected with Eimeria tenella

We evaluated the effects of dietary supplementation with different preparations of probiotics on the performance of broiler chickens experimentally infected with 2 × 10(4) sporulated oocysts of Eimeria tenella at 14 days of age. Three hundred, day-old, Cobb-500 chicks, as hatched, were separated into 10 equal groups with three replicates. Two of the groups, one challenged with E. tenella oocysts and the other not, were given a basal diet and served as controls without medication. The other challenged groups were given the anticoccidial lasalocid (60 mg/kg) or Enterococcus faecium (5 × 10(8) or 5 × 10(9)cfu/kg feed), Bifidobacterium animalis (5 × 10(8)cfu/kg feed), Lactobacillus reuteri (5 × 10(8)cfu/kg feed), Bacillus subtilis (5 × 10(8)cfu/kg feed), or a multi-species probiotic mix at 5 × 10(8) or 5 × 10(9)cfu/kg feed, respectively. The trial lasted 6 weeks. Individual body weight, feed intake per pen and feed conversion ratio values were recorded weekly, along with the extent of bloody diarrhea, excreta oocyst numbers and bird mortality. Caecal lesions were assessed and intestinal samples were taken for histopathological and bacteriological evaluation from ileum and caecum. Overall growth performance of chickens fed the multi-species probiotic mix at both levels was higher (P<0.05) compared to the infected control. Overall oocyst shedding was lowest (P<0.05) in the lasalocid supplemented group. Villous height was higher (P<0.05) in Bacillus supplemented groups compared to infected controls. The Lactobacillus supplemented group had the highest (P<0.05) numbers of both Lactobacillus and Bifidobacterium in ileum and caecum. In conclusion, dietary probiotics are promising for further investigation on improving intestinal health and growth performance of broiler chickens experimentally challenged with E. tenella.     

接种不同剂量E.tenella对雏鸡血清抗氧化酶的影响

同一日龄雏鸡接种不同剂量的柔嫩艾美尔球虫,研究接种不同剂量柔嫩艾美尔球虫对雏鸡血清抗氧化酶的影响。将60只健康鸡随机分为6组(n=10),分别接种0、0.5×104、1×104、2×104、4×104和8×104个孢子化卵囊。接种后7 d颈静脉采血,分离血清,测定血清脂质过氧化物(MDA)含量和超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-PX)、过氧化氢酶(CAT)活性。结果表明雏鸡感染不同剂量的柔嫩艾美尔球虫后,按接种剂量从低到高,血清中MDA含量先降低后升高变化差异极显著(P0.01);血清SOD活性先升高后降低,差异极显著(P0.01);血清GSH-PX活性先升高后降低,差异极显著(P0.01);血清CAT活性较对照组极显著性降低(P0.01)。不同剂量柔嫩艾美尔球虫感染对雏鸡血清MDA含量和SOD、GSH-PX、CAT活性有一定的影响。