Escherichia coli multiplication and lesions in the respiratory tract of chickens inoculated with infectious bronchitis virus and/or E. coli.

Escherichia coli numbers and histopathological changes were studied in the respiratory tract of line 151 chickens intranasally inoculated with infectious bronchitis virus (IBV) and/or virulent E. coli; this line is highly susceptible to IBV. Chickens inoculated with IBV alone showed increased numbers of E. coli in the trachea and had tracheitis, airsacculitis, and bronchiolitis. One of 17 chickens inoculated with IBV alone died with fibrinopurulent serositis. Chickens inoculated with IBV and E. coli had more severe and persistent respiratory lesions than those inoculated with IBV alone. E. coli was isolated from tracheas of chickens inoculated with IBV and E. coli more frequently than from chickens inoculated with IBV alone. In this group, 14 of 27 chickens died with tracheal plugs or with fibrinopurulent serositis. There was neither increased numbers of E. coli nor significant lesions in the respiratory tract of the group inoculated with E. coli alone. These results suggest that IBV may facilitate E. coli invasion into the lower respiratory tract of the chicken.     

Growth of Salmonella typhimurium in the caecum of gnotobiotic chickens with Eimeria tenella

Gnotobiotic chickens infected with Eimeria tenella (5 X 10(4) oocysts per bird) received an oral inoculation of 100 Salmonella typhimurium two, four, six or eight days after coccidial infection at four days old. When S typhimurium was given two or four days after E tenella infection, S typhimurium counts in the caecal contents were similar to the counts in birds infected with S typhimurium alone. When S typhimurium was given six or eight days after E tenella infection, counts of the organism were significantly greater than with S typhimurium infection alone. There were no differences in the number of chickens positive for S typhimurium in the caecal contents, bile, liver and spleen between the two groups.     

Blackhead disease (Histomonas meleagridis) aggravated in broiler chickens by concurrent infection with cecal coccidiosis (Eimeria tenella).

The effect of concurrent cecal coccidiosis infections on severity of Histomonas meleagridis (blackhead disease) in chickens was investigated in a series of experiments. Cecal lesions from H. meleagridis were severe in all inoculated control groups and did not appear to be affected by the introduction of Eimeria tenella infection. However, the severity of liver lesions and number of birds positive for liver lesions of H. meleagridis increased significantly with the presence of E. tenella. The increase was similar when 10(3) or 10(4) oocysts of E. tenella were given and was the same when oocysts were given at the same time as H. meleagridis or 4 days prior. The liver lesions increased directly as doses of H. meleagridis increased from 7.5 x 10(3) cells to 30, 100, or 300 x 10(3) when E. tenella was given along with H. melelagridis but not when H. meleagridis was given alone. Administration of a live coccidiosis vaccine containing very low levels of E. tenella also gave a significant boost to liver lesions but at a much lower level than that observed with larger doses of E. tenella. The positive relationship between infections of cecal coccidiosis and H. meleagridis in chickens suggests that such dual exposure may contribute to increased clinical outbreaks of blackhead disease in chickens under field conditions.     

鸡柔嫩艾美耳球虫SO7基因的原核表达及动物免疫试验

为了研究鸡柔嫩艾美耳球虫SO7基因原核表达重组蛋白的免疫效力,分析其在鸡抗柔嫩艾美耳球虫感染中对鸡平均增重、抗球虫指数(ACI)、减少盲肠病变和卵囊数量中的作用,从柔嫩艾美耳球虫孢子化卵囊中提取总RNA作为模板,用RT-PCR扩增出SO7基因片段,再应用DNA重组技术将SO7基因克隆到原核表达载体pET32a(+)中并用IPTG诱导表达。SDS-PAGE结果显示,pET32a(+)-SO7表达的目的蛋白约为40ku,主要以包涵体形式存在。用纯化的重组蛋白进行动物免疫试验显示,SO7重组蛋白在鸡抗柔嫩艾美耳球虫感染中有较好的免疫保护作用,能够显著提高鸡柔嫩艾美耳球虫感染鸡的平均增重和抗球虫指数(ACI),对减少盲肠病变和卵囊数量也有部分作用。     

Effect of Eimeria tenella infection on the caecal population of lincomycin-resistant Clostridium perfringens introduced into chickens

The effect of Eimeria tenella infection on the caecal population of lincomycin-resistant Clostridium perfringens (KGW 851) newly introduced into chickens was studied. Four groups of chickens consisted of: (1) Uninoculated controls; (2) inoculated with C perfringens (KGW 851); (3) inoculated with E tenella; and (4) inoculated with C perfringens (KGW 851) followed by E tenella. Five chickens in each group were necropsied on each of days 0, 3, 5, 7, 10 and 14 after the E tenella inoculation. The mean total C perfringens counts in the caecal contents increased at five days after E tenella inoculation and reached maximum counts at seven days after the inoculation. Also lincomycin-resistant C perfringens readily established itself at approximately one 10th of the total C perfringens population in the presence or absence of E tenella infection.     

实验性鸡大肠杆菌病病理学动态变化

用致病性大肠杆菌O18分离株和／或低致病性禽流感病毒（Mildly pathogenic avian influenza virus ,MPAIV)接种10－12日龄SPF鸡。在接种后1－96h进行临床症状与大体病理变化、组织学观察发现：大肠杆菌接种组、MPAIV接种组和健康接种组除扑杀鸡外未见鸡死亡，MPAIV与大肠杆菌混合接种组除扑杀鸡外死亡率为24％。混合接种组的病变比大肠杆菌接种组出现的时间早，恢复也慢，各脏器的病理变化更严重。MPAIV主要引起各实质器官的坏死，结果表明，大肠杆菌经气管内接种后试验鸡主要表现为呼吸道的炎症反应；MPAVI可使鸡大肠杆菌病严重化。     

实验性鸡大肠杆菌病的超微动态病理变化

10～ 12日龄 SPF鸡 180只 ,随机分为 4组 ,分别气管内注射致病性大肠杆菌 O18分离株 (大肠杆菌接种组 )、低致病性禽流感病毒 (mildly pathogenic avian influenza virus,MPAIV) H9N2株 (MPAIV接种组 )、先接种 MPAIV再接种大肠杆菌 (混合接种组 ) ,并设健康对照组 ,分别于接种后不同时间 ,取气管、肺、气囊、胸腺、法氏囊、脾、肝和肾组织 ,制作超薄切片 ,电镜观察。结果表明 ,MPAIV与大肠杆菌混合接种组比大肠杆菌接种组出现病变的时间早、恢复慢 ;在大肠杆菌接种组 ,接种后 3h气囊上皮和间质细胞中都可见典型的大肠杆菌 ;在混合接种组 ,接种后 3h,气囊间质细胞的吞噬泡中可见多个 MPAIV粒子。由此认为 ,MPAIV可使鸡大肠杆菌病严重化 ,大肠杆菌对 MPAIV的入侵和在鸡体内的复制可能有促进作用。     

Efficacy of decoquinate against drug sensitive laboratory strains of Eimeria tenella and field isolates of Eimeria spp. in broiler chickens in China

The efficacy of decoquinate against Eimeria infections in broiler chickens was evaluated using two drug sensitive laboratory strains of Eimeria tenella and 20 field isolates of Eimeria spp. collected from farms in China where various anticoccidials (including maduramicin) had been used. Decoquinate (20-40 ppm in feed) and maduramicin (5 ppm) were efficacious against E. tenella laboratory strains, but decoquinate more so than maduramicin. Body weight gains of E. tenella infected chickens were significantly improved, and caecal lesions were prevented, by feeding either decoquinate or maduramicin. Decoquinate also prevented oocyst production, but maduramicin did not. Most (18/20) Eimeria field isolates were resistant to maduramicin, judged by oocyst production; decoquinate at > or =20 ppm completely controlled all 20 field isolates. Decoquinate has potential value as a broiler anticoccidial in China and other countries where it has not been previously used.     

Pathology of spontaneous colibacillosis in a broiler flock

Forty-eight of 134 chickens collected from a flock on a broiler farm were diagnosed pathologically and microbiologically to have colibacillosis. Both acute septicemia (seven birds, 1 to 36 days old) and subacute serositis (41 birds, 5 to 57 days old) were found. The former consisted of necrosis with fibrinous exudates in the ellipsoids and lymphoid follicles of the spleen, and fibrinous thrombi in sinusoids of the liver with occasional necrosis of hepatic cells. The latter had fibrinopurulent inflammation with granulomatous changes in the serosal tissues--including the epicardium, pericardium, and hepatic peritoneal sac--accompanied by septicemic lesions in the spleen and liver. Respiratory lesions (airsacculitis, pneumonia, and tracheitis) were noted in most chickens affected with acute septicemia and subacute serositis. Degenerative changes also were observed in the bursa of Fabricius.     

Comparison of disease susceptibility and subclass-specific antibody response in SC and FP chickens experimentally inoculated with Eimeria tenella, E. acervulina, or E. maxima

Susceptibility to disease and the subclass-specific antibody response to Eimeria tenella, E. acervulina, and E. maxima were compared in two inbred strains of chickens, FP (B15B21) and SC (B2B2). FP strain was more susceptible to coccidiosis than SC chickens based on oocyst production, lesion score, and clinical signs. FP chickens infected with E. tenella had more severe cecal lesions and a significantly lower hematocrit level than SC chickens. FP chickens infected with E. acervulina excreted five times as many oocysts at 6 days postinfection as SC and showed a 71% reduction in plasma carotenoid level compared with controls (56% reduction in SC chickens). Body-weight change did not correlate with other signs of disease. Both SC and FP chickens produced high levels of serum IgM and IgG and biliary IgA. Although SC chickens had a slightly higher antibody response than FP chickens at 7 days postinoculation, both strains maintained high levels of IgM, IgG, and IgA for a prolonged period post primary inoculation. Although SC and FP chickens show different disease susceptibility to coccidiosis, they demonstrate similar antibody response.     

Coccidiosis: a radiological study of sulphaquinoxaline distribution in infected and uninfected chickens

Using scintillation counting and autoradiographical techniques, the whole-body distribution in week-old uninfected chickens of the anticoccidial agent sulphaquinoxaline (SQ) labelled with ³⁵S was established at various time intervals after a single oral dose either alone or following continuous in-feed medication with unlabelled SQ, and after a single intravenous dose. The distribution was also established in chickens infected with the coccidia Eimeria acervulina or E. tenella , after a single oral dose of radiolabelled SQ administered either alone or following continuous in-feed medication with unlabelled SQ, as for uninfected chicks. In all uninfected chicks, SQ was rapidly absorbed from the gut and was distributed to all tissues. It appeared at high concentrations in the bile and kidneys 0.5 h after dosing. In chickens that had previously received unlabelled SQ in the diet, a radiolabelled dose maintained steadier tissue concentrations than the sharp rise and fall detected after a single oral dose. Intravenous dosing of uninfected chicks showed that SQ was secreted by the crop, gizzard and caecal epithelia into their lumina. Infection with E. acervulina or E. tenella coincided with an overall 3.5-fold sustained increase of SQ concentration in chick tissues. An updated hypothesis including these new observations for the anticoccidial mode of action of SQ in chickens is expounded.     

Adhesion of bacteria to the cecal mucosal surface of conventional and germ-free chickens infected with Eimeria tenella.

When Salmonella typhimurium and Clostridium perfringens were tested in conventional chickens, larger numbers of S typhimurium and C perfringens adhered to Eimeria tenella-infected ceca than to uninfected ceca. In germ-free chickens, S typhimurium and C perfringens adhered to the E tenella-infected cecal mucosa more than to the uninfected cecal mucosa, but fewer Bacteroides vulgatus and Bifidobacterium thermophilum adhered to the E tenella-infected ceca than to the uninfected ceca. Many bacteria adhered to the lesions caused by E tenella as observed by scanning electron microscopy. On the basis of our findings, we suggest that infection with E tenella upsets the balance of competitive adherence of bacteria, allowing more colonization of S typhimurium and C perfringens.     

Interaction of aflatoxin with Eimeria tenella infection and monensin in young broiler chickens.

Young broiler chicks inoculated with Eimeria tenella and given a diet containing 2.5 mug aflatoxin/g had significantly higher mortality than birds with aflatoxicosis or coccidiosis alone or uninoculated controls. This effect was seen even when a light coccidial infection alone did not increase mortality or cause weight depression. In addition this higher mortality with the combination began earleir and occurred at a higher rate than did mortality from aflatoxin or cecal coccidiosis alone. Dietary monensin sodium (99 umg/g) did not completely prevent mortality and weight depression when aflatoxin and E. tenella were in combination. Aflatoxin and E. tenella singly significantly depressed three-week body weights; however, the depression was most severe when the two were in combination. Both dietary aflatoxin and E. tenella significantly reduced hemoglobin, packed cell volume, and plasma pigmentation, and in combination resulted in more severely reduced hemoglobin, packed cell volume, and plasma pigmentation. Coccidial lesion scores were significantly less for the combination of E. tenella and aflatoxicosis than for coccidiosis alone. This atypical response of the ceca to E. tenella in the presence of dietary aflatoxin was characterized by less distended ceca, very little coagulated blood in the ceca, and apparently more profuse cecal hemorrhage.     

Drug resistance in coccidia: a robenidine-resistant strain of eimeria tenella.

Tests for resistance to a recently introduced anticoccidial drug, robenidine, were performed on a strain of Eimeria tenella recovered from broiler chickens infected with acute cecal coccidiosis. The strain was identified previously and again confirmed by the timing of mortality of the infected chickens (five to six days postinfection), the appearance of lesions and parasites in ceca only, and the measurements (21.6 +/- 1.9 micron X 19.8 +/- 2.4 micron) of the oocysts. The tests showed that the strain of E. tenella could establish infections in chickens medicated with fourfold the recommended level of robenidine in feed but no oocysts could be recovered from the feces of infected chickens when the level of drug was increased to eightfold. The decrease in mortality, in the severity of lesions and oocyst output and the increase in the average weight gain of infected chickens followed closely the increase in the level of robenidine in feed. In contrast, no infections were found in chickens infected with a sensitive strain of E. tenella and maintained on feed mixed with the recommended level of robenidine. This and other findings discussed here show that resistance to robenidine is developing in coccidia commonly found in broiler houses.     

Possibilities of bacteremia and toxemia in death of chickens infected with Eimeria tenella

The possibility that bacteremia and toxemia were the causes of death in cases of cecal coccidiosis was investigated. Germ-free and ordinary chickens with microflora were inoculated with sporulated oocysts of Eimeria tenella. At 5 days postinoculation, cecal lesions in ordinary chickens were more severe than those in germ-free ones. Cardiac blood, spleen, and liver were examined in ordinary chickens for bacteremia and endotoxemia, and small numbers of bacteria were recovered from both infected and uninfected birds. Endotoxin levels in plasma of E. tenella-infected birds were low and not different from the levels of uninfected controls. To examine unknown toxic factors, a large volume of serum from infected chickens was injected intravenously into uninfected birds. No significant clinical signs were observed. It is concluded that the intestinal bacteria increase the severity of coccidial lesions without bacteremia and toxemia.     

Response to Eimeria tenella of chickens selected for high or low antibody response and differing in haplotypes at the major histocompatibility complex.

Sublines of chickens selected for high antibody (HA) or low antibody (LA) response that differed at the major histocompatibility complex (MHC) were tested for response to Eimeria tenella. In Expt. 1, the first exposure to E. tenella was natural (in floor pens), and chicks were challenged orally 21 days later with 0, 928, or 1855 oocysts. In Expt. 2, chicks were reared in wire-floored batteries, vaccinated orally with 928 oocysts, and challenged orally 12 days later with 15,844 oocysts. Corticosterone (20 mg/kg) was mixed with feed from 24 hr before vaccination to 120 hr after vaccination in Expt. 2. In Expt. 1, LA chicks had more-severe cecal lesions but gained relatively more body weight after challenge than did HA chicks. In Expt. 2, cecal lesions were least severe in HA chicks that had been fed corticosterone, most severe in LA chicks fed corticosterone, and intermediate in chicks that were not fed corticosterone. No differences in response to E. tenella occurred as a result of haplotypes at the MHC.     

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.     

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.     

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.     

柔嫩艾美耳球虫感染对肉仔鸡粪便水分的影响

采用单因子试验设计法,测定肉仔鸡感染柔嫩艾美耳球虫(E.tenella)后对粪便中水分变化的影响。试验表明:感染柔嫩艾美耳球虫后,鸡的生产性能受到显著的影响(P<0.05)。粪便中水分含量高于未感染组,感染后4d粪便中水分含量呈上升趋势,研究认为,粪便中水分含量的增加是球虫感染后的鸡的病理性变化的一种生理反映。在临床上,可将鸡粪便中水分含量的变化作为评判鸡感染球虫的参考指标之一。