Natural infection and transmission of a retrovirus closely related to myeloblastosis-associated virus type 1 in egg-type chickens

Myeloblastosis-associated virus type 1 (MAV-1) is an exogenous avian retrovirus with oncogenic potential. MAV-1 was detected in young chicks hatching from eggs produced by an experimental genetic line of egg-type chickens. Transmissibility of MAV-1 had not been documented previously. This investigation was intended to partially characterize the virus involved and to study its transmissibility and oncogenicity in naturally and contact-infected chickens. Commercially produced white and brown layer pullets free of exogenous avian leukosis viruses were commingled at hatch with naturally MAV-1-infected chickens. The original MAV-1-infected chickens were discarded after approximately 8 wk, and the contact-exposed chickens were maintained in isolation for 36 wk. Young specific-pathogen-free (SPF) single comb white leghorn chickens were added to the group to study possible horizontal transmission of MAV-1 in young chickens. Upon weekly virus isolation attempts, MAV-1 was readily isolated from the contact-exposed white layers but not from the brown layers between 36 and 53 wk of age (18 wk in total). Three-week-old SPF chickens were readily infected with MAV-1 by contact as early as 1 wk postexposure. Throughout 22 hatches derived from the white and brown MAV-1-contact-exposed layers (between 36 and 53 wk of age), MAV-1 was frequently detected in the white layer progeny, whereas the virus was seldom isolated from the progeny produced by the brown layers during the same 18-wk period. MAV-1 induced a persistent infection in some of the SPF chickens that were exposed by contact at 3 wk of age. Gross tumors were not detected in any of the originally infected experimental chickens at 8 wk of age, in the contact-exposed brown or white layers at the termination of the study at 53 wks of age, or in the contact-exposed SPF chickens at the end of the study at 12 wk of age. Exogenous avian leukosis-related viruses may still be detected in egg-type chickens, emphasizing the importance of thorough screening before incorporation of experimental genetic material into commercial genetic lines of egg-type chickens.     

Effect of breeder vaccination on immunization of progeny against Newcastle disease

Two experiments were conducted to determine the effect of breeder vaccination program and maternal antibody on the efficacy of Newcastle disease immunization of 1-day-old chicks. Experimental protocol was the same for both. In the first experiment, broilers were from breeders that were 32 weeks old, and in the second experiment, broilers were from breeders 50 weeks old. Breeders received three live Newcastle disease virus (NDV) vaccines and either a killed vaccine at 18 weeks or continual live boosting at 60-to-70-day intervals through lay. Broilers were vaccinated at 1 day of age with a commercial coarse-spray machine; they were bled, sera were examined for antibody against NDV, and broilers were challenged with virulent NDV at 2, 4, and 6 weeks of age. In the first experiment, maternal antibody was higher in chicks from the younger breeders given the inactivated vaccine, and in the second experiment maternal antibody was higher in chicks from older breeders given continual live vaccines. Higher antibody in 1-day-old broilers resulted in fewer vaccine-induced reactions, less vaccine virus shed, and decreased duration of vaccine-induced immunity from coarse-spray vaccination.     

Vaccination of 1-day-old chicks with fowlpox virus by the aerosol, drinking water, or cutaneous routes

Administration of 10(4) mean cell-culture infectious dose (CCID50) per ml of a plaque-purified derivative of a commercial fowlpox virus (FPV) vaccine to 1-day-old chicks by aerosol or drinking water gave inconsistent serological responses and little evidence of protective immunity. In contrast, cutaneous vaccination with the same preparation protected against challenge with virulent FPV at 4 weeks of age. Administration of the vaccine at a concentration of 10(6) CCID50 per ml by the drinking-water route was as effective as conventional cutaneous vaccination in terms of the serological response in an enzyme-linked immuno-sorbent assay and in terms of protection against challenge. Drinking-water vaccination at 2 days of age was no more effective than vaccination on day 1, and oral dosing with the vaccine was less effective than incorporation of the vaccine in the drinking water. It was concluded that 1-day-old chicks may be vaccinated against fowlpox by the drinking-water route if the vaccine contains a sufficiently high concentration of virus.     

Antibody response to and maternal immunity from an experimental psittacine beak and feather disease vaccine.

Adult umbrella cockatoos, Moluccan cockatoos, African grey parrots, and a yellow-headed Amazon parrot were inoculated IM or SC with beta-propiolactone-treated psittacine beak and feather disease (PBFD) virus. Thirty- to 45-day-old African grey parrot, umbrella cockatoo, and sulphur-crested cockatoo chicks also were vaccinated with the same inoculum. The hemagglutination inhibition (HI) and agar-gel diffusion tests were used to assay for post-vaccination development of anti-PBFD virus antibodies. All adult vaccinates seroconverted and had increases in HI and precipitating antibodies. The vaccinated chicks had increased concentrations of HI antibodies, but precipitating antibodies could not be detected. To demonstrate that chicks from vaccinated hens are protected from PBFD virus challenge, 3 African grey parrot chicks and 2 umbrella cockatoo chicks from vaccinated hens and 1 African grey parrot chick and 1 umbrella cockatoo chick from nonvaccinated hens were exposed to purified PBFD virus. Chicks from the vaccinated hens remained clinically normal during the 50-day test period. Chicks from the nonvaccinated hens developed clinical and histologic lesions of PBFD. Infected tissues from these birds were confirmed to contain viral antigen, using immunohistochemical staining techniques. The PBFD virus was recovered from the affected birds. These findings indicate that adult and 30- to 45-day-old psittacine birds will seroconvert following vaccination with beta-propiolactone-treated PBFD virus. Also, hens inoculated with beta-propiolactone-treated PBFD virus produce chicks that are, at least temporarily, resistant to virus challenge.     

Efficacy and safety of a cell-culture live virus vaccine for hemorrhagic enteritis of turkeys: laboratory studies

Poults free from hemorrhagic enteritis (HE) antibody were vaccinated by gavage at 1 day or 2 weeks of age with a live HE vaccine virus that had been propagated in a Marek's disease (MD)-induced B-lymphoblastoid cell line of turkey origin. Vaccinated and unvaccinated poults were challenged with a virulent HE virus at various times postvaccination. One hundred tissue-culture-infectious doses of the vaccine virus per poult were sufficient to induce a serological response as well as to protect poults against HE lesions and mortality. Vaccinated poults were protected against the disease as early as 1 week and as late as 8 weeks PV. The vaccine was efficacious by several routes of application. The vaccine virus spread horizontally from vaccinated to contact-exposed poults, as indicated by seroconversion and resistance of contact-exposed poults to challenge. The vaccine had no detectable harmful effects on the humoral immune response to particulate antigens or on weight gain of vaccinated poults. The vaccine proved to be free from MD virus, as indicated by the absence of MD lesions and antibody in 8-week-old chickens inoculated intra-abdominally with the vaccine at hatching. These findings indicate that the cell-culture-propagated HE vaccine is efficacious and safe.     

Genetic susceptibility of indigenous chicks to subgroup A Rous sarcoma virus inoculated via the chorioallantoic membrane.

An investigation was made using chicks of two Indian indigenous breeds of fowl, Kadaknath and Aseel, to ascertain genetic resistance to infection by Rous sarcoma virus of subgroup A. A standard inoculation dose of 0.2 ml virus containing 1000 pock forming units ml-1 was injected via the chorioallantoic membrane (CAM) into the 11-day-old embryos that were subsequently hatched. The sensitivity of the two indigenous breeds was compared with the highly susceptible exotic White Leghorn (WL) strain maintained in the laboratory. The Kadaknath breed was about three-fold and Assel, about six-fold less sensitive than the WL strain, indicating superiority of the indigenous breeds over the exotic breed of fowl. Most of the CAM-susceptible chicks died of liver tumour (LT) and most of the CAM-resistant chicks survived. However, conversely associated tumour phenotype subclass chicks, i.e. CAM-susceptible LT-negative chicks that survived and CAM-resistant LT-positive chicks that died, occurred consistently in the three breeds of fowl. Nevertheless, the overall survival potential of Kadaknath chicks measured up to 8 weeks post-hatching was greater than that of Aseel chicks. Neither transformation of embryonic tissue prior to hatching nor the visceral metastasis including liver conformed with the degree of CAM-infection as measured by number of pocks on CAMs.     

Molecular identification of the Marek's disease virus vaccine strain CVI988 in vaccinated chickens

The study describes three polymerase chain reaction (PCR) systems for the CVI988 vaccine virus: the meq gene, the MDV BamHI-D/H 132 bp tandem repeat fragment and the MDV-gB gene. Whereas the PCR product of virulent MDV strains and of the CVI988 virus strain with the meq and the 132 bp primer sets differed for the two templates, the MDV-gB PCR products were similar. The sensitivity of the three PCRs was determined for the two templates: the CVI988 DNA was detected up to 2.48 plaque forming units, and a MDV-1 DNA, was amplified with the 132 bp primers up to the 10(-3) DNA dilution, and up to the 10(-2) with the MDV-gB and meq gene primers. As conventional detection for the CVI988 vaccine virus is by tissue culture, the aim was to analyse the feasibility of the molecular detection of the vaccine virus in the vaccinated chick. In two experimental trials employing specific pathogen free and commercial Lohmann chicks, respectively, the vaccine virus replicated to a limited extent; it was detected only in the spleen of up to 60% chicks at 2-4 weeks and in one chick at 3 weeks, respectively. The survey of three commercial Lohmann flocks, kept in biosecurity conditions, revealed the vaccine virus only in the spleen of 40% of 30-day-old chicks. The present study shows that CV1988 DNA is present in vaccinated chicks in a low quantity and it is difficult to detect directly from the chick, probably because vaccine viruses are latent in vivo. For an efficient detection it is pertinent to cultivate the vaccine virus on chicken embryo fibroblasts (CEF), as then the virus escapes the latent state, enters into the productive mode of replication, and a high viral copy number is produced.     

β-羟基-β-甲基丁酸对肉仔鸡生长性能和屠宰性能的影响

选择240只7日龄肉仔鸡按照体重相近原则随机分为5组,每组6个重复,每个重复8只,分别饲喂添加0%、0.01%、0.055%、0.1%、0.145% β-羟基-β-甲基丁酸(HMB)的试验日粮.采用三层笼养,试验期35 d.试验结果表明:(1)不同水平HMB对肉仔鸡生产性能的总体影响不显著.7～21日龄时,与对照组相比,0.145% HMB组内仔鸡料重比降低6.06%(P＞0.05);0.1%和O.145%HMB肥组肉仔鸡死亡率低于对照组水平.22～42日龄时,0.145%HMB组肉仔鸡采食量与对照组相比降低9.94%,各处理组体增重和料重比差异不显著(P＞0.05);7～42日龄时.0.145%HMB组肉仔鸡采食量与对照组相比降低9.03%(P＜0.05),各处理问体增重和料重比无显著差异,但0.145%HMB组肉鸡料重比最小.0.1%和0.145%HMB组肉仔鸡死亡率低于对照组和其他处理组.(2)肉仔鸡的屠宰性能包括屠宰率、半净膛、全净膛率、胸肌率、腿肌率和腹脂率在各处理问无显著差异(P＞0.05),0.145%HMB组肉仔鸡腿肌率较对照组提高3.74%(P＝0.075).可见,在7～42日龄内仔鸡日粮中添加适量的HMB可显著降低肉仔鸡采食量.而体增重和饲料效率无显著变化,并有促进肉仔鸡腿部肌肉生长发育的趋势.     

A paramyxovirus of serotype 3 isolated from African and Australian finches

Two virus isolates were obtained from exotic finches (Ortygospiza atricollis and Poephila cincta) suffering from apathy, diarrhea, conjunctivitis, and dysphagia. The isolates were identified as paramyxoviruses based on their multiplication characteristics in embryonating chicken eggs, chicken embryo fibroblasts, and chicken embryo kidney cell cultures, on morphology upon electron microscopy, and on other biological properties. Both isolates were serologically related to the reference strain of the paramyxovirus serotype 3. Intravenous infection of 42-day-old chicks resulted in no clinical signs, but intracerebral infection of 1-day-old chicks resulted in mortality and intracerebral pathogenicity indices of 0.25 to 0.35. Of five finches from various species inoculated with isolate 840/85, three remained clinically healthy through 6 weeks, but two died: one (Poephila cincta) 5 days postinoculation after showing nervous distress, and the other (Amandava amandava) suddenly 42 days postinoculation.     

Effect of infectious bursal disease on the response of chickens to Mycoplasma synoviae, Newcastle disease virus, and infectious bronchitis virus.

At 35 days of age, chickens which as 1-day-old chicks were inoculated with the infectious bursal disease virus (IBDV) had significantly lower antibody titers against Mycoplasma synoviae, Newcastle disease virus, and infectious bronchitis virus than did those never inoculated with IBDV. The IBDV also had a marked effect on the development of air-sac lesions. Birds infected with IBDV that were later inoculated with M synoviae (day 14), Newcastle disease virus (days 14 and 28) experienced an increased incidence and greater seversity of airsacculitis than did chicks which were not exposed to IBDV.     

Experimental infection in specific-pathogen-free chicks with avian reovirus and avian nephritis virus isolated from broiler chicks showing runting syndrome

Avian reovirus (ARV) and avian nephritis virus (ANV) were individually isolated from runty 10-day-old broiler chicks. The ARV isolate, IR-R, the ANV isolate, IR-N, and the reference strain of ANV, G-4260, were inoculated orally into 1-day-old chicks of two specific-pathogen-free (SPF) chicken lines, 151 and PDL-1. Growth retardation without the presence of gross lesions was clearly observed at 7 and 14 days postinoculation (PI) in chicks of both lines inoculated with the IR-R strain. On the other hand, in chicks inoculated with IR-N strain, growth retardation was observed only in the chicks of line 151 at 7 and 14 days PI. Microscopically, nephritis was observed in both chicken lines at 7 and 14 days PI. When chicks that were inoculated with the IR-N strain at 1 day of age were inoculated with the IR-R strain at 3 days of age, growth retardation was observed in the chicks of line PDL-1 at 10 and 17 days PI. However, the growth retardation was less severe than in the group receiving a single inoculation of the IR-R strain.     

Pathogenesis of two strains of avian paramyxovirus serotype 2, Yucaipa and Bangor, in chickens and turkeys

Nine serologic types of avian paramyxovirus (APMV) have been recognized. Newcastle disease virus (APMV-1) is the most extensively characterized virus, while relatively little information is available for the other APMV serotypes. In the present study, we examined the pathogenicity of two strains of APMV-2, Yucaipa and Bangor, in 9-day-old embryonated chicken eggs, 1-day-old specific-pathogen-free (SPF) chicks, and 4-wk-old SPF chickens and turkeys. The mean death time in 9-day-old embryonated chicken eggs was more than 168 hr for both strains, and their intracerebral pathogenicity index (ICPI) was zero, indicating that these viruses are nonpathogenic in chickens. When inoculated intracerebrally in 1-day-old chicks, neither strain caused disease or replicated detectably in the brain. This suggests that the zero ICPI value of APMV-2 reflects the inability of the virus to grow in neural cells. Groups of twelve 4-wk-old SPF chickens and turkeys were inoculated oculonasally with either strain, and three birds per group were euthanatized on days 2, 4, 6, and 14 postinoculation for analysis. There were no overt clinical signs of illnesses, although all birds seroconverted by day 6. The viruses were isolated predominantly from the respiratory and alimentary tracts. Immunohistochemistry studies also showed the presence of a large amount of viral antigens in epithelial linings of respiratory and alimentary tracts. There also was evidence of systemic spread even though the cleavage site of the viral fusion glycoprotein does not contain the canonical furin protease cleavage site.     

Shedding of lymphoid leukosis virus in chickens following contact exposure and vaccination

Chickens contact-exposed to lymphoid leukosis virus at various ages up to 32 weeks responded with relatively high rates of infection as determined by the presence of neutralizing antibody. Virus shedding as determined by cloacal swab and albumen testing occurred in 7 of 8 groups of such chickens, but the incidence was 10% or less and sporadic. Vaccination of chickens immediately before exposure with a low pathogenicity virus of subgroup A at 8 weeks of age did not eliminate subsequent shedding.     

Genetic variations in maternal transfer and immune responsiveness to infectious bursal disease virus

The immune responsiveness to infectious bursal disease virus (IBDV) in four native and crossbred chicken lines was compared. ELISA IBDV antibody titers in hen serum samples, yolk from matched eggs and sera from matched 1-day-old chicks from each chicken line with an identical vaccination program were measured, and plotted. There was considerable variation between lines in the measured IBDV specific antibodies, in vaccinated parent hens and in the amounts of inherited maternally derived antibodies in both yolk and progeny chicks. Differences in ratios of the inherited antibody level from hen to 1-day-old chicks were also found among different chicken lines. Breed differences in regressions of IBDV antibody levels in yolk to that of hen or progeny chicks' sera were also found, so prediction of serum titer of hen and/or progeny chicks from yolk are varied among chicken lines.     

Glycosylation of the envelope protein of West Nile Virus affects its replication in chicks

Birds are important for the transmission of West Nile virus (WNV) in nature, but the significance of the potential N-linked glycosylation at position 154 in the WNV envelope (E) protein with regard to viral replication in young chickens has not been assessed. In this study, the effect of glycosylation of the WNV E protein on viral pathogenicity in birds was investigated using young domestic chicks. A higher viral load was detected in the blood and the peripheral organs, particularly the hearts, of 2-day-old chicks inoculated with a glycosylated WNV variant compared to those inoculated with the nonglycosylated variant. There was no significant difference in the neutralizing antibody titers and cytokine expression profiles in chickens inoculated with the glycosylated and the nonglycosylated WNV variants. In contrast, no virus w as detected in the blood and the tissues of 3-wk-old chicks, although the host immune response was induced to similar levels as in the 2-day-old chicks. These data indicate the utility of young domestic chicks as an animal model of WNV infection; they also indicate that glycosylation of the E protein of WNV enhances multiplication in the blood and peripheral organs, which is associated with the strong pathogenicity of WNV in birds.     

不同来源禽网状内皮增生病病毒株的致病性比较

本研究旨在探讨不同来源的禽网状内皮增生病病毒(REV)对1日龄SPF鸡的致病性.用包括分子克隆化病毒REV-C99在内的不同来源的6个REV株人工感染1日龄SPF鸡,以禽流感病毒灭活疫苗(H9-AIV、H5-AIV)与新城疫病毒灭活疫苗(NDV)免疫后HI抗体滴度的测定结果为指标,比较不同REV株的致病性.结果表明,分别用3000TCID_(50)·只~(-1)的剂量人工感染1日龄SPF鸡,6个REV感染组与对照组相比,H9-AIV、H5-AIV与NDV免疫后4、5与6周的HI抗体水平均极显著降低(P<0.01).但6个REV感染组之间差异不显著(P>0.05).研究结果显示,不同来源REV株感染1日龄SPF鸡后均造成生长迟缓,并对体液免疫反应有明显的抑制作用,同时,这也揭示出REV感染可能是免疫失败的重要原因之一.     

Experimental infection of specific-pathogen-free chickens with avian rotaviruses

One-to-70-day-old specific-pathogen-free chickens were infected with rotaviruses isolated from chickens, turkeys, and pheasants. Chicks infected at 1 or 7 days of age remained largely free of infection: virus could be isolated from a few chicks of these ages, but virus antigen could not be detected in the epithelial cells of the intestinal villi. Treating the virus with trypsin before inoculation did not markedly enhance virus infectivity. The same batches of virus successfully infected chickens at 56 and 70 days of age: virus could be isolated between 1 and 5 days postinfection, and virus antigen was detected in the epithelial cells of the intestinal villi. Rotaviruses isolated from pheasants were infectious for 49-day-old chickens. All infections in the older chickens remained subclinical.     

Experimental avian paramyxovirus serotype-3 infection in chickens and turkeys

Avian paramyxoviruses (APMV) are divided into nine serotypes. Newcastle disease virus (APMV-1) is the most extensively characterized, while relatively little information is available for the other APMV serotypes. In the present study, we examined the pathogenicity of two divergent strains of APMV-3, Netherlands and Wisconsin, in (i) 9-day-old embryonated chicken eggs, (ii) 1-day-old specific pathogen free (SPF) chicks and turkeys, and (iii) 2-week-old SPF chickens and turkeys. The mean death time in 9-day-old embryonated chicken eggs was 112 h for APMV-3 strain Netherlands and > 168 h for strain Wisconsin. The intracerebral pathogenicity index in 1-day-old chicks for strain Netherlands was 0.39 and for strain Wisconsin was zero. Thus, both strains are lentogenic. Both the strains replicated well in brain tissue when inoculated intracerebrally in 1-day-old SPF chicks, but without causing death. Mild respiratory disease signs were observed in 1-day-old chickens and turkeys when inoculated through oculonasal route with either strain. There were no overt signs of illness in 2-weeks-old chickens and turkeys by either strain, although all the birds seroconverted after infection. The viruses were isolated predominantly from brain, lungs, spleens, trachea, pancreas and kidney. Immunohistochemistry studies also showed the presence of large amount of viral antigens in both epithelial and sub-epithelial lining of respiratory and alimentary tracts. Our result suggests systemic spread of APMV-3 even though the viral fusion glycoprotein does not contain the canonical furin proteases cleavage site. Furthermore, there was little or no disease despite systemic viral spread and abundant viral replication in all the tissues tested.     

Pathologically and serologically different avian nephritis virus isolates implicated in etiology of baby chick nephropathy.

Three virus isolates (WG-3, -4, and -5) from chicks affected by baby chick nephropathy were orally inoculated into 1-day-old specific-pathogen-free chicks of lines PDL-1 and 15I. Additional chicks were orally inoculated with avian nephritis virus (ANV) strain G-4260. Chicks inoculated with isolates WG-3, -4, and -5 died between 2 and 6 days postinoculation (PI), with mortality ranging from 0% to 53.3%. Pathological findings in the dead chicks included nephrosis in chicks inoculated with WG-3, -4, and -5, and nephritis and visceral urate deposition in chicks inoculated with G-4260. The stability of the WG-5 isolate, as well as the size of the particles and the nucleic acid type, were also similar to those of the G-4260 strain. All of the examined chicks inoculated with WG-3, -4, and -5 had interstitial nephritis at 14 days PI. Therefore, the three virus isolates were considered to be ANV. However, there was no serological relationship between the isolates and ANV (G-4260 and M-8 strains).     

鸡肾型传染性支气管炎组织病理学变化规律的研究

为制造疾病模型便于防治鸡肾型传支。将1日龄健康雏鸡70只分为A、B、C 3组(试验组、对照组和感染组),试验组人工接种IBV肾型毒株,感染组晚1 d放入试验组内混养,常规饲养管理,每日4次观察接毒后的临床症状,同时定期剖检病鸡和病死鸡,取多脏器固定于10%福尔马林液中、石蜡切片、HE染色,显微镜下观察组织病理学变化,按严重程度标+~++++后记录并标明曲线。结果表明,在整个试验过程中肾脏的病理变化最为严重并且持续时间较长,从第2天开始出现病变,并持续到第20天;其次是肺脏,病变出现在第5~16天;肝脏和肺脏的病变时间相同只是程度轻一些;心脏的病变出现在第6~13天。1 d后,感染组也出现相似的病理变化,说明此病的传染性很强。