Use of lipopolysaccharide (LPS) as a positive control for the evaluation of immunopotentiating drug candidates in experimental avian colibacillosis models

The aetiologic agent of avian colibacillosis is Escherichia coli. Colibacillosis is a disease that causes mortality and production performance problems in chickens which results in economic losses for the poultry industry. It will be increasingly important for scientists to identify novel solutions that can be implemented which will provide poultry producers with a tool to manage this economically important disease. The purpose of this investigation was to determine whether lipopolysaccharide (LPS) could be used as a positive control to evaluate novel chemistries for immunopotentiator activity in battery or floor-pen avian colibacillosis models in chickens. In the battery study, subcutaneous administration of LPS to one-day-old broiler cockerels caused a significant reduction (P < 0.003) in all parameters of colibacillosis evaluated, i.e. mean air sac lesion scores, per cent air sac lesions, E. coli re-isolation and per cent mortality. However, in the floor-pen study, subcutaneous administration to one-day-old broiler chicks resulted in a numerical, but not statistically significant reduction (P < 0.1) in mortality associated with colibacillosis. These data indicate that LPS can be used as a positive control to evaluate the efficacy of immunopotentiator drug candidates in avian colibacillosis models.     

Immunogenicity of an oil-emulsified Escherichia coli bacterin against heterologous challenge

Immunogenicity of an oil-emulsified Escherichia coli bacterin against heterologous challenge was investigated. In Expts. 1 and 2, chickens were vaccinated with E. coli serotype O1 bacterin and challenged with E. coli serotype O2 (Expt. 1) and O78 (Expt. 2). Positive control chickens were not vaccinated but challenged with E. coli serotype O2 or O78; unvaccinated unchallenged chickens served as negative controls. When challenged with E. coli serotype O2, unvaccinated chickens showed a higher morbidity than vaccinated chickens. There was no mortality in either group. Although average gross lesion scores were generally higher in the unvaccinated chickens, they were not significantly different (P greater than or equal to 0.05) from those in the vaccinated chickens. In Expt. 2, morbidity was slightly higher in the unvaccinated challenged chickens. No mortality occurred in either group. There was no significant difference (P greater than or equal to 0.05) between vaccinated and unvaccinated chickens in average gross lesion scores. In general, E. coli recovery was higher in the unvaccinated challenged chickens, being highest in the air sacs followed by the liver, heart blood, and pericardial sacs. There was no morbidity, mortality, or gross lesions in the unvaccinated unchallenged chickens. No E. coli was recovered from the tissues cultured. The results of these laboratory trials revealed that an oilemulsified monovalent E. coli bacterin did not protect chickens against other E. coli serotypes associated with colibacillosis.     

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。     

Experimental Escherichia coli respiratory infection in broilers

This study determined optimal conditions for experimental reproduction of colibacillosis by aerosol administration of avian pathogenic Escherichia coli to 2-to-4-wk-old broiler chickens. The basic model for reproducing disease was intranasal administration of approximately 10(4) mean embryo infectious dose of infectious bronchitis virus (IBV) followed by aerosol administration of an 02 or an 078 strain of E. coli in a Horsfall unit (100 ml of a suspension of 10(9) colony-forming units/ml over 40 min). Scores were assigned to groups of infected chickens on the basis of deaths; frequency and severity of lesions in the air sacs, liver and heart; and recovery of the challenge E. coli 6 days post-E. coli infection. An interval of 4 days between the IBV and E. coli challenges was best whether the chickens received the IBV at 8 or 20 days of age. Typically, 50%-80% of the chickens developed airsacculitis and 0 to 29% of the chickens developed pericarditis or perihepatitis, with little or no mortality. Escherichia coli alone resulted in no deaths and 0 to 20% airsacculitis, but these percentages increased to 0 to 5% and 52%-60% when the E. coli aerosol was administered through a cone-shaped chamber. Administration of IBV alone failed to induce lesions. Recovery of the challenge E. coli from chickens did not correlate well with lesions. On the basis of these data, administration of IBV to 20-day-old chickens followed 4 days later by exposure to an avian pathogenic E. coli reproduces avian colibacillosis with the low mortality, high percentage of airsacculitis, and low percentage of septicemic lesions characteristic of the conditions seen in the natural disease.     

Cellular defense of the avian respiratory system: protection against Escherichia coli airsacculitis by Pasteurella multocida-activated respiratory phagocytes

The concept of nonspecific cellular defense of the respiratory system of poultry against respiratory pathogens by "preventive activation" of avian respiratory phagocytes (ARPs) was tested in an in vivo protection trial. Chickens were stimulated intratracheally by Pasteurella multocida Choloral vaccine strain. Seven hours later, these and mock-inoculated control chickens were challenged with pathogenic Escherichia coli via the air-sac route. Stimulated chickens had a 25-fold-elevated number of ARPs compared with mock-inoculated control chickens. The proportion of active phagocytes and the phagocytic capacity of these cells was higher in the ARP populations of stimulated chickens than in the ARP populations of control chickens. In vivo protection against E. coli air-sac infection was demonstrated by reduction of morbidity and mortality rates, diminished weight loss, and lower scores of gross and histopathological lesions of P. multocida-stimulated chickens compared with mock-inoculated controls.     

Isopathic and pluralist homeopathic treatment of commercial broilers with experimentally induced colibacillosis

This study sought to determine the efficacy of isopathic and pluralist homeopathic treatment of colibacillosis in broiler chickens and thereby contribute to the evaluation of homeopathy in general. In each of two experiments three groups of broilers, infected intratracheally at 8 days of age with E. coli (O78:K80), were treated with different combinations of homeopathic remedies. Control groups and an infected, doxycyline-treated group were included. Experiments differed only in the dose of E. coli. Efficacy of treatment was evaluated based on the parameters mortality, body weight gain and colibacillosis lesions. In both experiments doxycyline prevented mortality and reduced E. coli lesions and stunting. None of the homeopathically treated groups differed significantly with respect to any of the parameters from the non-medicated, infected control group. It is concluded that the results of this study do not justify use of these homeopathic remedies for treatment of colibacillosis in broilers. Furthermore, no significant effects of this homeopathic treatment were established.     

Clinical and microbial efficacy of antimicrobial treatments of experimental avian colibacillosis

The clinical and microbial efficacy of antimicrobial treatments of avian colibacillosis was studied, using an experimental model on chickens previously inoculated with multiresistant commensal Escherichia coli strains. One E. coli with pMG252 plasmid containing bla(FOX5) and qnrA1 genes and another E. coli with pMG298 plasmid containing bla(CTX-M15) and qnrB1 genes were first orally inoculated to chickens Both isolates were also resistant to chloramphenicol, sulphamethoxazole, trimethoprim, streptomycin, gentamicin, kanamycin, and tetracycline. The birds were then experimentally infected with an avian pathogenic E. coli (APEC), via the air sac. Treatments (oxytetracycline (OTC), trimethoprim-sulfadimethoxin (SXT), amoxicillin (AMX) or enrofloxacin (ENR) were then offered at the therapeutic doses. Symptoms, lesions in dead or sacrificed birds, and isolation and characterization of APEC from internal organs were studied. Results showed that OTC, SXT or ENR treatments could control the pathology. AMX worsened the disease, possibly due to endotoxin shock. All APEC re-isolated from internal organs showed the same antimicrobial susceptibility as the APEC inoculated strain, except for one APEC isolate from an infected OTC-treated bird, which acquired tetracycline resistance only, and one APEC isolate recovered from the air sacs of a chicken in the infected SXT-treated group, which acquired the pMG252 plasmid and became multi-resistant. Thus three antimicrobials could control the disease but the experimental model enabled, to our knowledge, the first observation of plasmid transfer from a bacterium of the intestinal tract to a pathogenic isolate from the respiratory tract.     

Mass medication in reducing shipping fever-bovine respiratory disease complex in highly stressed calves

One thousand and eighty-five newly received, stressed calves were used in studies to determine the effectiveness of certain mass medication procedures for reducing morbidity from shipping fever-bovine respiratory disease complex. In two experiments, im injections of oxytetracycline at 11 mg/kg body wt for 3 successive days reduced treatment days/calf purchased 21 (P less then .05) and 31% (P less than .05). Oral administration of 150 mg of sulfadimethoxine/kg body wt reduced treatment days/calf purchased 20 (P less than .05) and 54% (P less than .05) in the same two experiments. When sulfadimethoxine followed oxytetracycline on the third injection day an 81% reduction in treatment days/calf purchased was obtained, indicating an additive effect of the two drugs. The use of long acting oxytetracycline and sustained release sulfadimethoxine at the time of processing resulted in a 90% reduction in treatment days/calf purchased (P less than .01) and required only one handling of the calves for mass medication purposes.     

Dietary induction of sudden death syndrome in broiler breeders

Sudden death syndrome in broiler breeders at the point of lay was a serious problem in Australia for 18 months. This condition was reproduced by a combination of dietary and management practices. Chickens fed a diet low in potassium, phosphorus, protein, and energy had plasma potassium and phosphorus levels significantly lower than the levels seen in the controls. The plasma values of the test breeders were similar to levels seen in field cases. There was a significant difference (P less than 0.01) in the venous acid-base balance between the test chickens and controls. Test chickens were alkalotic and had a 6% mortality rate between 18 and 30 weeks of age.     

Immunogenic potency of an oil-emulsified Escherichia coli bacterin

Immunogenicity of an oil-emulsified Escherichia coli (O1:K1) bacterin with an aqueous-phase-to-oil-phase ratio of 1:4 was evaluated in chickens. Chickens were vaccinated subcutaneously with 0.5 ml of the bacterin at 4 and 6 weeks of age. At 8 weeks, the vaccinated chickens and unvaccinated controls were challenged via air sacs with 10(4) colony-forming units (CFU) of homologous E. coli. Vaccinated chickens were protected against active respiratory infection in that they (a) gained body weights comparable to those in unvaccinated, unchallenged chickens, (b) suffered no morbidity or mortality, (c) had gross lesions so mild that the scored values were comparable statistically to the 0 lesion scores of the negative controls, and (d) did not yield E. coli when their heart blood, pericardial sacs, livers, and air sacs were cultured. Unvaccinated challenged chickens had severe respiratory distress, suffered 36% mortality, and had average air sac, pericardial sac, and liver lesion scores significantly (P less than or equal to 0.05) different from both the vaccinated and negative control chickens. Also, the challenge strain of E. coli only was isolated from the affected tissues of 5 of 14 chickens. Protection against active respiratory infection was again demonstrated in a second experiment, though the challenge dose was 1.06 X 10(6) CFU of E. coli. The immunity, however, was partially overcome, as the vaccinated chickens gained less body weight and the scored values for lesions in the air sacs, pericardial sacs, and livers were significantly higher than those of the negative controls (P less than or equal to 0.05).     

不同小麦日粮对肉仔鸡肉质、脂肪酸合成酶mRNA与脂蛋白脂肪酶mRNA表达的影响

本文旨在研究不同小麦日粮对肉仔鸡生长、肉质及脂肪酸合成酶(FAS)mRNA与脂蛋白脂肪酶(LPL)mRNA表达的影响。选用300只艾维因肉仔鸡公雏,随机分为5组,每组4个重复,每个重复15只鸡,5组分别饲喂玉米日粮和4种水溶性阿拉伯木聚糖(WSAX)含量不同的小麦日粮,试验期为6周。结果表明:①随着小麦日粮中WSAX含量的增加,肉仔鸡的生长明显降低;②小麦日粮显著增加了肌肉持水力,并随小麦WSAX含量增加而升高的趋势显著(P<0.01),剪切力也随之增加(P<0.05)。饲喂小麦日粮的肉仔鸡其肉色显著低于玉米日粮组(P<0.01),各小麦日粮组间差异不显著。当小麦WSAX含量较高时,小麦日粮可以显著降低肉仔鸡肌肉脂肪和胆固醇含量(P<0.05)。各处理间胸肌、腿肌含水量无差异(P>0.05);③小麦日粮显著降低FASmRNA在肉仔鸡肝中的表达量(P<0.01),并且随日粮WSAX含量增加而表达水平降低的趋势明显。各处理间LPLmRNA表达量的差异不显著(P>0.05)。本研究表明:①当小麦中WSAX含量较低时,对肉仔鸡增重和肉质的影响较小,是玉米理想的替代品;②当小麦中WASX含量较高,则会严重影响肉仔鸡的增重、肉质及FASmRNA在肝脏中的表达,不宜在肉仔鸡日粮中大量添加。     

热应激试验性大肠杆菌病肉鸡心肺病理组织学的观察

研究了37℃热应激条件下试验性大肠杆菌病肉鸡心脏和肺脏的病理组织学变化。结果表明:热应激条件下试验性大肠杆菌病肉鸡心脏的病理性损伤较单纯感染大肠杆菌病肉鸡严重,而肺脏损害较轻。     

Field trials with a bivalent vaccine (HVT and SB-1) against Marek's disease

White leghorn chickens on five farms were given a bivalent Marek's disease (MD) vaccine consisting of turkey herpesvirus (HVT) and SB-1 (a nononcogenic MD virus); other chickens received only HVT. The farms had histories of "vaccination failures," presumably owing to an exceptionally virulent challenge MD virus. The bivalent vaccine uniformly protected chickens better than HVT alone between 12 and 16-20 weeks of age, when serious MD losses occurred. During that period, total mortality in groups given both viruses ranged from 0.39 to 1.26% (mean 0.86%), whereas that in HVT-vaccinated groups not exposed to SB-1 varied from 1.92 to 7.44% (mean 3.43%). Chickens in pens or rows with close contact to those given bivalent vaccine also had low MD mortality rates (0.46-1.06%, mean 0.77%), probably from the spread of SB-1.     

A comparison of prophylactic efficacy of tilmicosin and a new formulation of oxytetracycline in feedlot calves

Two replicated-pen field studies were performed under commercial feedlot conditions in western Canada to compare the administration of long-acting oxytetracycline at 30 mg/kg body weight (BW) versus tilmicosin at 10 mg/kg BW to feedlot calves upon arrival at the feedlot. Ten thousand nine hundred and eighty-nine, recently weaned, auction market derived, crossbred beef steer and bull calves were randomly allocated upon arrival at the feedlot to one of 2 experimental groups as follows: oxytetracycline, which received intramuscular long-acting oxytetracycline (300 mg/mL formulation) at a rate of 30 mg/kg BW; or tilmicosin, which received subcutaneous tilmicosin (300 mg/mL formulation) at a rate of 10 mg/kg BW. There were 20 pens in each experimental group. In Study 1 and in the combined analysis, the initial undifferentiated fever (UF) treatment rate was significantly (P < 0.05) higher in the oxytetracycline group as compared with the tilmicosin group. There were no significant (P > or = 0.05) differences in first UF relapse, second UF relapse, third UF relapse, overall chronicity, overall rail, overall mortality, bovine respiratory disease (BRD) mortality, hemophilosis mortality, arthritis mortality, or miscellaneous mortality rates between the experimental groups in either study or in the combined analysis. In addition, there were no significant (P > or = 0.05) differences in initial weight, final weight, weight gain, days on feed, daily dry matter intake, average daily gain, or the dry matter intake to gain ratio between the experimental groups in either study or in the combined analyses. In the economic analysis, there was a net economic advantage of $5.22 CDN per animal in the oxytetracycline group, due to a lower prophylactic cost, even though the UF therapeutic cost was higher.     

Colonization of Escherichia coli in young turkeys and chickens.

In order to investigate the possibility of pathogenic Escherichia coli penetrating the bloodstream via the intestinal mucosa in normal and stressed turkeys and chickens, birds were inoculated orally with the bacteria or exposed environmentally to it. Immediately after hatch, intestines contained a substantial number of coliform bacteria that increased with time. In orally infected turkeys, the pathogenic bacteria (nalidixic-acid-resistant O78) replaced 10%-50% of the native coliform flora but could not be isolated from the trachea or blood. Environmentally exposed groups exhibited pathogenic bacteria in intestines but not in blood. Stressing of exposed turkeys resulted in isolation of the pathogenic bacteria from blood and even spleen. In orally infected broiler chickens, stress resulted in bacteremia and mortality. Chickens that were exposed to pathogenic bacteria at a young age and showed no mortality or morbidity demonstrated no detrimental effects due to challenge with the same pathogenic bacteria later in life. Stress seems to cause penetration of the pathogenic bacteria into the bloodstream, which in turn can cause severe disease and mortality.     

Safety,immunogenicity, and efficacy of two Escherichia coli cya crp mutants as vaccines for broilers

Attenuated derivatives (delta cya delta crp mutants) of an O2 and an O78 avian septicemic Escherichia coli strain were used to immunize broiler chickens by spray to determine the safety, immunogenicity, and efficacy of the derivatives in single- and double-dose regimens. In the safety and immunogenicity studies, groups of 10 chickens were vaccinated by spray (droplet size approximately 20 microm) with the parent E. coli, the mutant organisms, or phosphate-buffered saline (PBS) at 14 days of age and euthanatised 21 days later. There was no deaths or gross pathologic finding in any of the chickens immunized with the vaccine strains. Compared with the levels in chickens exposed to PBS, there were significantly higher levels of immunoglobulin (Ig) G antibody in serum and air sac washings and of IgA antibody in air sac washings in response to the virulent parent strains than to the vaccine strains. In efficacy studies, chickens were immunized with the O2 or the O78 vaccine strain or PBS at day 14 and with the O2 vaccine strain or PBS at days 10 and 14 and challenged with the parent strain 10 days after the last vaccination. There was no significant difference in local IgA and IgG and serum IgG responses between vaccinated and control groups. Chickens vaccinated with the O2 strain, but not the O78 strain, had significantly lower air sac lesion scores compared with those of the unvaccinated groups in both single- and double-dose regimens. We conclude that the mutant O2 strain provided moderate protection against airsacculitis.     

卵黄性腹膜炎、鸡白血病对种鸡死亡率、产蛋率影响的初步调查

为了探讨卵黄性腹膜炎和鸡白血病对种鸡死亡率和产蛋率的影响，在2016年1月至2017年4月中旬，对江苏省常州市某种鸡场岭南黄鸡、花鸡（金茅花鸡）、草鸡（金茅草鸡）、黄脚麻鸡4个品种鸡的发病及死亡情况进行临床症状观察及病理剖解；同时记录鸡场每日的鸡死亡数、产蛋数、每日免疫和用药情况，分析不同时期的死亡率和产蛋率，比较不同品种鸡的产蛋率及产蛋高峰期产蛋率与正常蛋鸡产蛋高峰期产蛋率（参考值80%）之间的差异。从死亡鸡的剖解结果来看，腹腔内卵黄凝固黏连在肠道上，输卵管肿大，充满白色或淡黄色干酪样物，初步诊断为卵黄性腹膜炎；肝脏和脾脏肿大，肠系膜或胸壁有肿瘤状物体，初步诊断为鸡白血病。岭南黄鸡产蛋后期的死亡率极显著高于产蛋上升期和高峰期（P<0.01），黄脚麻鸡产蛋后期的死亡率极显著低于产蛋高峰期（P<0.01）；花鸡、草鸡、黄脚麻鸡产蛋高峰期产蛋率平均值均低于60%，与参考值（80%）差异极显著（P<0.01）。综合试验结果，卵黄性腹膜炎和鸡白血病对鸡场影响严重，导致鸡死亡率上升，产蛋率下降；抗应激药和营养药可以起到缓解预防的作用，但无法根治，尚无有效的治疗方法。     

恩诺沙星对鸡大肠杆菌病及葡萄球菌病的药效研究

就畜禽专用氟喹诺酮类抗菌药恩诺沙星对鸡人工感染大肠杆菌病及葡萄球菌病的药效进行了研究。结果表明，恩诺沙星、氯霉素对大肠杆菌Ｏ７８的ＭＩＣ分别为０．０５、１．６０ｍｇ／Ｌ；恩诺沙星、红霉素对金黄色葡萄球菌Ｃ５６０５株的ＭＩＣ分别为０．２、３．２ｍｇ／Ｌ。人工发病试验中，恩诺沙星内服以５、１０、２０ｍｇ／ｋｇ剂量或肌注以２．５、５、１０ｍｇ／ｋｇ剂量，每天给药２次，连用３ｄ，对鸡大肠杆菌病及葡萄球菌病均有较好的疗效，其中对大肠杆菌病的疗效明显优于氯霉素     

几种病毒与禽病原性大肠杆菌的人工联合感染

以2种剂量的低致病性禽流感病毒(lowly pathogenic avian influenza virus,LPAIV)，传染性支气管炎病毒（infectious bronchitis virus,IBV)疫苗株H120和H52，新城疫病毒（Newcastle disese viurs,NDV)Lasota株分别于气管内注射10日龄易感鸡，2d后，气管注射禽病原性大肠杆菌O37株（O78），连续观察5d，结果，除LPAIV单独感染组有6．25％的死亡率外，其余各病毒单独接种组均健活；大肠杆菌O37株单独接种组的死亡率为62．50％，较高剂量的LPAIV，IBV H120和H52，NDV Lasota株与大肠杆菌O37株有效强的协同致病作用，死亡率分别达到81.25%,100.00%,93.75%和87.50%，而较低剂量的上述病毒则无明显的协同作用，IBV，NDV疫苗株与大肠杆菌联合接种组的多数死亡鸡病程推迟。     

The effect of sodium chloride extract and commercial lipopolysaccharides of Escherichia coli and Salmonella typhimurium on chickens.

In chickens inoculated into the heart with a sodium chloride extract of Escherichia coli strain (serotype O2) isolated from a chicken with colibacillosis, characteristic hemorrhages into the anterior chamber of the eyes (hyphema) were found. Significant lesions were limited to the eyes. Cyclophosphamide-treated chickens were more sensitive to the extract than untreated chickens and hyphema was usually seen in association with hemorrhages of the iris. These activities were not reduced by heating the extract at 60 degrees C for one hour or by trypsin digestion. Chickens inoculated into the heart with commercial lipopolysaccharides of E. coli (serotypes O111:B4 and O55:B5) and Salmonella typhimurium showed similar lesions in the eyes as the chickens inoculated with the sodium chloride extract. These findings suggest that the endotoxin may induce hyphema in chickens.