Experimental studies on Marek's disease in Japanese quail (Coturnix coturnix japonica)

Day-old quails experimentally infected with Marek's disease (MD) virus of quail origin developed lymphoid tumors. The severity of the disease increased considerably with serial passage. Tumor transplants could be made with cells derived from gross tumors in skeletal muscles, spleen cells, and blood from MD-affected quails. After five to six serial transplants, the tumor could not be transplanted further. Marek's disease tumor-associated surface antigen (MATSA) was demonstrated in lymphoid cells of spleen and peripheral blood lymphocytes of MD-affected quails. The MATSA of quail differed from the MATSA of chicken. Chickens were susceptible to MD virus isolated and propagated in quails.     

Immune complex-mediated glomerulopathy in Marek's disease

Chickens infected with Marek's disease (MD) virus developed immune complex (IC)-mediated glomerulopathy. Fluorescent antibody staining technique using antichicken globulin and antichicken complement was used to demonstrate IC in the kidney glomeruli. During the initial stages of MDV infection, IC deposits were seen on the glomerular basement membrane, but subsequently the entire glomerulus was involved. Mesangial cells also had IC deposits. Chicken complement was demonstrated in the glomeruli which had IC deposits. The number of glomeruli with IC deposition was higher in tumor-bearing birds than in non-tumor-bearing birds. Histologically, kidney lesion were characterized by thickening of basement membrane and proliferation of mesangial cells. It is suggested that IC-mediated glomerulopathy might be one of the major causes of death in MD.     

Marek's disease in Japanese quails (Coturnix coturnix japonica): a study of natural cases

Marek's disease was observed in quails. Gross lesions were confined mostly to the spleen and liver. Microscopic lesions were commonly seen in spleen, proventriculus, liver, and duodenum. Skin, peripheral nerves, and other visceral organs were also involved. Of 123 quails examined, 39 had serum antibodies against Marek's disease. These antibodies were detected from 11 to 17 weeks of age; the highest incidence was recorded at 15 weeks. Feather follicular antigen detected in 30 of the 95 quails was comparable to that of chicken. The disease was experimentally reproduced in susceptible quails. Marek's-disease-tumor-associated surface antigens (MATSA) were demonstrated in the peripheral leukocytes and spleen cells of affected quails. The possible source of infection and its epidemiological importance are discussed.     

Pathogenicity and transmission of reticuloendotheliosis virus isolated from endangered prairie chickens

The pathogenicity and transmission of a field isolate of reticuloendotheliosis virus (REV) was studied using an experimental model in Japanese quail. Oncogenicity was also evaluated after inoculations in chickens and turkeys. The original REV (designated APC-566) was isolated from Attwater's prairie chickens (Tympanuchus cupido attwateri), an endangered wild avian species of the southern United States. The transmissibility of the REV isolate was studied in young naive Japanese quail in contact with experimentally infected quail. Vertical transmission was not detected by virus isolation and indirect immunofluorescence. Seroconversion was detected in few contact quails, suggesting horizontal transmission. The APC-566 isolate induced tumors beginning at 6 wk of age in quails infected as embryos. Most of the tumors detected in Japanese quail were lymphosarcomas, and 81% of these neoplasias contained CD3+ cells by immunoperoxidase. REV APC-566 was also oncogenic in chickens and turkeys infected at 1 day of age, with tumors appearing as early as 58 days after infection in chickens and at 13 wk of age in turkeys. This study was conducted in part as an attempt to understand the potential for pathogenicity and transmission of REV isolated from endangered avian species.     

Immunological aspects of Marek's disease in chickens with maternal antibodies]

The dynamics of the production of immunoprecipitation antibodies to Marek's disease virus was studied in the serum of chickens with maternal antibodies in relation to the occurrence of the immunoprecipitation antigens of Marek's disease virus in feather follicles. One-day-old chickens were infected by the contact method with Marek's disease virus. The first occurrence of immunoprecipitation antigen was detected on the 14th day after infection and this occurrence persisted throughout the experiment, i. e. until the 112th day after infection. The antibodies were first detected the 28th day after infection and their titre kept rising until the 98th day after infection. Immunoprecipitation antibodies and antigens of Marek's disease virus were detected in some tumorously changed kidneys. Immunoelectrophoretic examination revealed in the same kidneys immunoglobulins of the class IgY, IgA and beta-globulin. The slowest-migrating fraction of IgY, together with IgA, beta-globulin and C-reactive protein were detected in the skin extracts from infected poultry. Indirect haemagglutination enabled the detection of the presence of haemagglutination antibodies in rabbit immunoglobulin to the skin antigen of Marek's disease virus, and in avian immunoglobulin to the same virus. Haemagglutination antigen was revealed in the extract from tumorously changed kidneys.     

A study of Marek's disease in Japanese quails vaccinated with herpesvirus of turkeys

In a study of outbreaks of Marek's disease in quails, 220 adult quails vaccinated with herpesvirus of turkeys (HVT) were examined pathologically and serologically for Marek's disease (MD). Forty-three of the 220 quails exhibited microscopic lesions similar to those of chickens with MD. MD-virus-specific antigen was found in feather tips of 44 of the 220 birds, and the HVT-specific antibody was found in sera of 56 of the 220 birds by agar gel precipitation. There was a positive correlation between the incidence of lymphomatous changes and the presence of MD-virus-specific antigen, and there was a negative correlation between the incidence of lymphomatous changes and the presence of antibody against HVT on a flock basis.     

The occurrence of Marek's disease in genetically different groups of chickens

The occurrence of Marek's disease was studied under laboratory conditions in four genetically different groups of chickens (Brown Leghorn, F1 hybrids of the CB x IA inbred lines, pullets of the paternal branch of the grandparent stock of the Hybro meat type, and final hybrids of the White Hisex layer type) after infection with the Georgia strain of Marek's disease (MD) virus, used in two doses (1600 and 16,000 PFU per one bird). MD was diagnosed on the basis of the occurrence of macroscopic tumours; when these tumours were absent in birds which had died within 105 days from infection, the dead bodies were subjected to microscopic examination (peripheral nerves, gonads, skin, bursa of Fabricius, thymus, spleen). The size of the parenterally administered dose of the virus had no significant effect on mortality, occurrence of tumours and the MD virus in any of the groups of chickens tested. However, there was a time shift in the mortality curve in chickens infected with a lower dose. The significantly highest occurrence of MD was recorded in BrL chickens (100%). A somewhat lower occurrence of MD was recorded in the Hisex White chickens (87.8%) and in the CB x IA hybrids (73.8%). However, the dead CB x IA chickens had a higher occurrence of tumours (96.6%) than the Hisex White chickens (77.1%). The lowest MD occurrence was recorded in the pullets of the Hybro meat type (25.5%). The organ most frequently affected by tumours after infection of the birds with the Georgia strain was the liver (24.1%).     

Early cell-mediated immune responses to Marek's disease virus in two chicken lines with defined major histocompatibility complex antigens

N2a and P2a chickens, resistant and susceptible to Marek's disease (MD), respectively, were used to examine relationships between major histocompatibility complex (MHC)-restricted cytotoxic T lymphocytes (CTL) and natural killer (NK)-like cell activity with resistance to infection with Marek's disease virus (MDV). Ten-day-old chickens were infected with MDV and euthanatized at selected times to evaluate for NK cell and MHC-restricted cytotoxicity. The N2a MDV-infected chickens had an early cell-mediated immune response characterized by a sustained NK-like cytotoxicity that coincided with a measurable MHC-cytotoxicity that was lower than controls. Although MHC-restricted and NK cell cytotoxicity was demonstrated in P2a MDV-infected chickens at 8 dpi, both abruptly decreased and remained low for the remainder of the 20-day experiment. The critical time point that may determine the resistance to MD appears to be within the first 2 weeks post-infection. Improvement of the chicken NK cell activity may be a good candidate for both selection and immunomodulation MD control programs.     

Observations of experimentally induced quail bronchitis

Quail bronchitis was experimentally reproduced in captive-reared bobwhite quails (Colinus virginianus). Quails were inoculated with 10(6) mean tissue culture infective doses of quail bronchitis virus at 1,3,6, or 9 weeks of age by the intratracheal, intraperitoneal, or subcutaneous route. Clinical signs were minimal, but occasionally birds were ruffled, exhibited open-mouthed breathing, and developed "snicks." Mortality rates of quails inoculated at 1 or 3 weeks ranged from 7% to 87%. Quails inoculated at 6 or 9 weeks of age had mortality rates from 0% to 20%. Mean body weights of survivors that had been inoculated at 3 or 6 weeks were significantly less than those of controls (P less than 0.05). No significant differences in body weight were detected between quails inoculated at 1 or 9 weeks and their uninoculated controls. Antibodies to group I adenovirus were detected by agar gel precipitation in 87.5% of birds that survived infection.     

AUS in the prevention of Marek's disease

Aminoureidosulfone (AUS) was used as a feed additive to prevent mortality in Marek's disease (MD)-infected chickens in the laboratory. Chicks infected with MD virus (5000 plaque-forming units) at 1 day of age were given AUS in the feed from 1 day of age until the experiment was terminated. AUS, at all concentrations tested, reduced mortality due to Marek's disease. Chickens receiving AUS at the 0.002% level had a 4% mortality rate, those receiving 0.005% had 9% mortality, and those receiving 0.01% had 2% mortality; mortality rates in the respective controls were 44%, 49%, and 50%. However, lesion reduction in surviving chicks was minimal. When AUS was withdrawn from the feed, MD-induced mortality increased within two weeks; average body weight decreased also.     

The early pathogenesis in chicken inoculated with non-pathogenic serotype 2 Marek's disease virus.

A newly cloned serotype 2 Marek's disease virus (MDV), strain ML-6, was inoculated via the nasal cavity in specific-pathogen-free chicks to examine early virus replication and the expression of Marek's disease (MD)-related antigens. Following inoculation, viral intracellular antigens (VIAs) were detected in lymphoid organs (bursas and spleens) between 5 and 14 days post inoculation (PI), in feather follicles between 14 and 30 days PI, and in lungs at 3 days PI by the immunohistopathological staining of avidin-biotin-peroxidase complex method. But, very few VIAs were expressed in the thymuses between 5 and 14 days PI. However, MD tumor-associated surface antigens were not detected in any organs. Viruses were isolated from separated spleen cells at 14 and 30 days PI. Fluorescent antibodies of convalescent sera were also detected after 10 days PI. As most of the VIAs were detectable in B-cells in bursas and spleens. B-cells were considered to be the main first target cells for the serotype 2 MDV infection.     

MDV对雏鸡血浆MDA含量与SOD活性的影响

为探讨丙二醛(MDA)和超氧化物歧化酶(SOD)在雏鸡马立克病(MD)发病过程中的作用,研究了雏鸡在人工攻毒后血浆MDA含量与SOD活性的动态变化.结果表明,攻毒组接种马立克病病毒(MDV)后第14天,血浆中MDA的含量显著高于对照组(P＜0.05),第7天和第28天高于对照组,第56天时基本接近;血浆中SOD活性在第7天时对照组显著高于攻毒组(P＜0.05),第14天和第28天高于攻毒组,但差异不显著.表明雏鸡在感染MDV后血浆中MDA含量显著升高,SOD活性明显降低,对雏鸡MD的发病诊断有重要的指导作用.     

雏鸡马立克氏病强毒感染后脂质过氧化物的含量变化

１日龄雏鸡马立克氏病强毒（ｖＭＤＶ）人工感染马立克氏病（ＭＤ）发病率６２％,ＭＤ死亡率３４％;脾脏、胸腺、法氏囊、心脏、肝脏、肾脏、性腺脂质氧化物（ＬＰＯ）含量高于（Ｐ＞０．０５）或显著高于健康对照雏鸡（Ｐ＜０．０５）。     

Genetic characterization and susceptibility on poultry and mammal of H7N6 subtype avian influenza virus isolated in Japan in 2009

From February to March 2009, six strains of H7N6 subtype avian influenza virus were isolated from quails in three farms in Aichi prefecture in Japan. The isolates were shown to be low pathogenic for chicken by the examination performed using the "Manual of Standards for Diagnostic Tests and Vaccines" by World organisation for Animal Health (OIE). The deduced amino acid sequence at the cleavage site was PE (I/Q/L) PKRR (nucleotide sequences were cct gaa (a/c) (t/a) a cc (a/g) aaa aga aga), suggesting persistence in domestic poultry for some time. The direct putative ancestor strain could not be elucidated by phylogenetic analysis of all genome segments of the quail isolates. Diverged date from a putative common ancestor in a non-rooted phylogenetic tree among quail viruses was estimated between March 2002 and July 2004. Three putative N-linked glycosylation sites resided in the vicinity of the receptor binding pocket of HA1 region. They are considered to decrease the reactivity of neutralizing antibody against the virus. Experiments for the infectivity and pathogenicity of a quail strain to poultry indicated that the quail isolate had higher infectivity to quails than chickens and ducks. Direct and dust-borne and/or droplet-borne transmissions among quail were proven in quails with and without direct contact with experimentally infected quails. The virus is seldom transmitted among chickens either directly or indirectly, and indirect transmission from infected quails to chickens was not observed. The pathogenicity of the quail strain for mammalian, pig and mouse was low, although it could replicate in those animals.     

马立克氏病病毒感染鸡羽髓上皮细胞差异蛋白质组学研究

为鉴定鸡羽髓上皮细胞感染马立克氏病病毒(MDV)前后差异表达的蛋白,本研究以MDV强毒GA株人工感染SPF鸡,并通过双向电泳技术进行分析.结果显示:在病毒感染后4 d、7 d、14 d和21 d显著差异表达的蛋白点分别有2个、8个、25个和9个;而通过质谱技术鉴定出29种蛋白质,其中包括能量代谢相关蛋白、增殖和凋亡相关蛋白、细胞骨架蛋白、信号传导蛋白、转录相关蛋白、免疫相关蛋白和其他功能蛋白质.本实验首次对鸡羽髓上皮细胞感染MDV后各时期蛋白表达水平的变化进行研究,鉴定了多种差异表达蛋白质,为进一步揭示MDV与宿主的相互关系、感染性病毒粒子的成熟和致病机制提供了依据.     

Pathology of experimentally induced quail bronchitis

Bobwhite quails (Colinus virginianus) were inoculated with 10(6) mean tissue-culture infective dose of quail bronchitis virus at 1, 3, 6, or 9 weeks of age by intratracheal, intraperitoneal, or subcutaneous routes. Quails developed necrotizing tracheitis, proliferative and necrotizing bronchitis and pneumonia; multifocal necrotizing hepatitis; necrotizing splenitis, with or without hyperplasia of splenic mononuclear phagocytes; bursal lymphoid necrosis; and bursal atrophy. Lesions were more extensive and severe in quails inoculated at 1 or 3 weeks of age than in older quails. Large intranuclear inclusions, characteristic of adenovirus infection, were identified in trachea, lung, liver, and bursa of Fabricius. This is the first report of the histopathology of experimentally induced quail bronchitis.     

Delayed replication of Marek's disease virus following in ovo inoculation during late stages of embryonal development

Several oncogenic and non-oncogenic isolates of Marek's disease virus (MDV) were inoculated into embryonated eggs on embryonation day (ED) 16 to 18, and embryos or chicks hatching from inoculated eggs were examined for infectious virus and viral internal antigen (VIA) in lymphoid organs. There was no evidence of extensive replication of MDV in any of the embryonic tissues examined. Levels of VIA peaked 4-5 days after chicks hatched. This indicated that MDV remained inactive during embryonation and did not initiate pathogenic events until chicks hatched. Because HVT replicated rapidly in the embryo but MDV did not, in ovo inoculation of HVT simultaneously with oncogenic MDV or several days after MDV resulted in significant protection (P less than 0.025) of hatched chicks against Marek's disease (MD). Little protection was obtained if HVT was given simultaneously with MDV or after MDV to chicks already hatched. The relative susceptibility of the embryo to extensive replication of the vaccine virus but not the challenge virus apparently accounted for protection against MD in chicks hatching from dually infected eggs.     

Protecting chickens against Marek's disease under laboratory conditions by the administration of Marvak, a Czechoslovak produced vaccine and vaccine imported from East Germany

Laboratory trials were conducted to compare the efficiency of two vaccines against Marek's disease (MD) on the basis of the dynamics of post-vaccination viraemia and by means of a challenge test. Two groups of the HX-SL hybrid layers, each containing 40 birds, were vaccinated with the MARVAK cellular vaccine manufactured in Czechoslovakia (100 PFU per chick) or with the Insel Riems vaccine manufactured in the GDR (1000 PFU per chick). In weekly intervals, half the birds of each group were tested for post-vaccination viraemia and the other half, including 20 non-vaccinated birds, were infected by contact with the pathogenic virus of MD from the second day of age. The viraemia in the birds vaccinated with MARVAK culminated in the third week of age when it reached the average value of 25 PFU/10(7) leucocytes; later on, by the eighth week of age, it decreased to 0.62 PFU/10(7) leucocytes. The maximum number of viraemic chicks was found in the third and fourth week of age (80%); on an average it ranged from 60%. The average viraemia in the chickens treated with the vaccine from the GDR ranged from 1.6 to 3.6 PFU/10(7) leucocytes in the second to eighth week of age. A 100% positivity of the tested birds was found in the sixth week of age; it ranged around 70% on an average. In all the challenged groups of birds the precipitation antigen of Marek's disease was detected in feather follicles already 14 weeks from contact infection.(ABSTRACT TRUNCATED AT 250 WORDS)     

Influence of thymic hormone on cell-mediated and humoral immune responses in Marek's disease

A reduction in the secretion of thymic hormones, and in particular thymulin, can be demonstrated in chickens following the thymic atrophy induced by Marek's disease virus (MDV) infection. In very sensitive histocompatible (B13/B13) chickens inoculated with the HPRS-16 strain of MDV at 10 days of age, treatment with synthetic thymulin by daily subcutaneous injection failed to modify the time course of Marek's disease (MD) and did not prevent the development of macroscopic tumors. No effect was noted on the levels of neutralizing anti-viral antibodies. Nevertheless, thymulin treatment resulted in significant suppression of the cellular immune response 4-6 weeks post-inoculation, monitored by splenic cytotoxicity against MD-specific and natural killer-sensitive lymphoblastoid cell lines.     

Experimental Aspergillus fumigatus infection in quails and results of treatment with itraconazole

This study was performed to investigate (i). the clinical, histopathological and biochemical changes in quails (Coturnix coturnix japonica) with experimentally induced aspergillosis; and (ii). the efficiency of itraconazole treatment on these infected birds. A total of 18021-day-old male quails was randomly divided into three groups (control, infected untreated and infected treated), each containing 60. The experimental infection was set by intratracheal inoculation of 0.2 ml inoculum of Aspergillus fumigatus (CBS 113.26 strain) consisting of approximately 2.7 x 106 spores/ml. Two days after the inoculation, general clinical signs of aspergillosis in the respiratory tract were observed. In the histopathological examination, caseous foci were found in lungs, trachea and on airsacs. All quails died in the infected untreated group. Aspergillus fumigatus was isolated from the various organs of all dead quails. There was no significant change in serum aspartate aminotransferase (AST) and gamma-glutamyltransferase (GGT) activities in infected untreated birds compared with controls. However, alanine aminotransferase (ALT) activity, albumin and calcium levels, and albumin/globulin (A/G) ratio were lower while phosphorus and globulin levels were higher in the infected untreated group than in controls. Each quail in the infected treated group was given 10 mg/kg/day itraconazole via drinking water for 7 days immediately after the first clinical findings. Although all quails died in the infected untreated group, 41 quails survived in the itraconazole treatment group. Biochemical values also returned approximately to the control levels after the treatment. The conclusion was drawn that aspergillosis in the quails might cause economical losses because of high mortality. Oral itraconazole treatment of aspergillosis might lower the mortality rate in quails.