The effect of cyadox and virginiamycin on Marek's disease in chickens

Chickens infected with highly virulent Marek's disease virus were administered in the course of 56-day fattening cyadox at a dose of 20 mg per kg feed mixture (MC group). and virginiamycin at a dose of 10 mg per kg feed mixture (MV group). The weight gains on the 56th day and dressing percentage were highest, in comparison with the control group (711.11 g and 60.62%), in the virginiamycin-treated group. An investigation into the basic chemical composition of meat revealed lower contents of dry matter and proteins in the virginiamycin-treated group (26.9% and 22.76%) if compared with the control group of chickens which were also infected with Marek's disease virus. A biometrical examination of the organs showed a statistically significant decrease in heart weight in the cyadox-treated group, increase in liver and heart weight and intestine shortening in the virginiamycin-treated group, in comparison with the control group. Pathologico-anatomical changes typical of Marek's disease were recorded in a randomly selected number of chickens in 33.3% in the control group, in 28.5% in the cyadox-treated group and in 50.0% in the virginiamycin-treated group.     

Effects of vaccine dose, virus challenge dose and interval from vaccination to challenge on protection of broiler chickens against Marek's disease virus challenge

OBJECTIVES: To examine the effects of varying the doses of turkey herpesvirus (HVT) vaccine and Marek's disease virus (MDV) challenge at two intervals after vaccination on the protection of chickens against challenge with MDV. DESIGN AND PROCEDURE: Experiment 1, a dose response study, consisted of 11 doses of HVT vaccine administered at hatch followed by challenge with 100 plaque forming units (pfu) of MDV 5 days post vaccination. Experiment 2, a 2 x 6 x 2 factorial design, included two HVT vaccine types, six different doses of HVT vaccine and 50 pfu and 200 pfu of MDV challenge 2 days post vaccination. All chickens were reared up to day 56 post challenge when all survivors were killed humanely. Dead and killed chickens were examined for gross MD tumours. RESULTS: Experiment 1 showed a significant positive linear relationship between dose of HVT vaccine and protective index in chickens challenged 5 days post vaccination. However the range of protective index observed was limited. In Experiment 2 neither HVT vaccine provided significant protection at any dose. There was no significant effect of vaccine type or MDV challenge dose on overall protection against challenge. Chickens challenged with 200 pfu of MDV had significantly higher mortality and MD incidence than those with 50 pfu. CONCLUSIONS: HVT vaccine dose had a significant impact on protective index, but vaccination to challenge interval appeared to have greater impact on the protective efficacy of vaccination. A fourfold increase in challenge dose increased mortality rate and incidence of MD.     

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.     

Comparative susceptibility of Marek's disease cell lines to chicken infectious anemia virus

Chicken infectious anemia virus (CIAV) is known to infect and replicate in various Marek's disease chicken cell lines (MDCCs) derived from Marek's disease (MD) tumors. One line, MDCC-MSB1, has been the substrate used in most studies. We compared a total of 26 MDCCs, including two sublines of MDCC-MSB1, MSB1 (L) and MSB1 (S), four other MD tumor-derived lines, and 20 lines derived from MD virus-induced local lesions, for susceptibility to the Cux-1 and CIA-1 strains of CIAV. The cell lines represented six phenotypic groups of T cells based on the expression of CD4, CD8, and TCR-2 and -3 surface markers. Susceptibility was measured by the number of cells positive for viral antigen in immunofluorescence (IF) tests at 3-10 days postinfection. No clear-cut differences were found in susceptibility related to phenotype, although CD4-/8+ lines and CD4-/8- lines might be more susceptible than CD4+/8- lines. However, several individual lines were more susceptible to Cux-1 than the two MSB1 sublines tested. Contrary to an earlier report, cells of MDCC-CU147, a CD8+, TCR3+, local-lesion derived line, were found to be susceptible to CIA-1. In fact, CU147 was distinguished by very high susceptibility to both CIAV strains. In direct comparisons with MSB1, CU147 detected approximately 10-fold lower doses of virus. Also, virus spread was faster (P < 0.05) in CU147 than in MSB1 and other lines. Results from polymerase chain reaction (PCR) tests to detect infection in titrations were in general agreement with IF test results although PCR detected infection in a few terminal dilution cultures that were negative by IF.     

Indications of immunodepression in chickens infected with various strains of Marek's disease virus

The effect of infection by various strains of Marek's disease virus (MDV) on the immune function of 3-week-old chickens was examined. MDV strains of low (CU-2, RB-7) and high (RB-3, MD-5, and MD-11) pathogenicity were compared with prototype JM-10 strain of moderate pathogenicity. Mortality, whole body weight, relative weights of lymphoid organs, histopathology, humoral antibody responses to thymus-dependent and -independent antigens, and in vitro lymphocyte responses to mitogen stimulation were investigated at 1, 2, and 3 weeks postinfection. MDV strains of high pathogenicity significantly depressed responses at 3 weeks postinfection, seeming to indicate the ability of these viruses to induce severe immunodepression. However, the fact that the moderately pathogenic and even some of the low-pathogenicity strains induced immunodepression suggests that other viral mechanisms are also important in its determination.     

Susceptibility of adult chickens, with and without prior vaccination, to challenge with Marek's disease virus

Marek's disease (MD) outbreaks can occur in previously healthy adult layer or breeder flocks. However, it is not clear whether such outbreaks are caused by recent challenge with highly virulent (vv and vv+) strains of MD virus (MDV; i. e., new infection hypothesis) or by exacerbation of an earlier MDV infection (i. e., old infection hypothesis). To discriminate between these hypotheses, adult White Leghorn chickens of laboratory strains or commercial crosses with or without prior vaccination or MDV exposure were challenged at 18-102 wk of age with highly virulent MDVs, and lesion responses were measured. Horizontal transmission was studied in one trial. Challenge of adult chickens, which were free from prior MDV vaccination or exposure, with highly virulent MDV strains induced transient paralysis or tumors in 60%-100% of 29 groups (mean = 91%), and horizontal spread of virus was detected. The magnitude of the response was similar to that induced by challenge at 3 wk of age. In contrast, comparable challenge of adult chickens, which had been vaccinated or exposed to MDV early in life, induced transient paralysis or tumors in 0%-6% of 12 groups (mean = 0. 5%), although some birds showed limited virologic evidence of infection and transmission of the virus to contacts. The MD responses were influenced by the virulence of the challenge virus strain, and to a lesser extent by virus dose and route of exposure. Strong inflammatory lesions were induced in the brain and nerves of adult specific pathogen-free (SPF) chickens at 9-15 days after infection. The low susceptibility of previously vaccinated and exposed groups to challenge at > or =18 wk of age suggests that late outbreaks of MD in commercial flocks are not likely a result of recent challenge alone and that additional factors could be involved.     

Mortality of one-week-old chickens during naturally occurring Marek's disease virus infection

Marek's disease (MD) is a disease of chickens that occurs worldwide and has serious economic consequences. MD can present as one of several forms, with the most commonly occurring forms being the lymphoproliferative diseases. Under experimental conditions, an early mortality syndrome has been recognized following infection by some but not all strains of MD virus (MDV). This is the first report of a confirmed case of mortality due to naturally occurring MDV infection in 1-week-old, nonvaccinated, chickens. Necrotizing lesions were observed in the bursa of Fabricius, lung, duodenum, jejunum, and proventriculus, and large intranuclear inclusion bodies were a striking feature in tissues with lesions in all birds. Immunohistochemical staining for the pp38 protein of MDV revealed abundant pp38 antigen in the affected tissues, confirming the presence of MDV within the lesions. PCR yielded an amplicon with 97% homology to the meq gene of MDV. No evidence of co-infection by either of the immunosuppressive agents chicken anemia virus and infectious bursal disease virus was detected.     

不同马立克氏病抗性鸡攻毒后IL-18基因的差异表达

本试验致力于评价鸡白细胞介素-18(IL-18)基因的表达模式与马立克氏病抗性遗传性状的相关性。MDV-1分别感染7日龄普通品系和抗性品系的霞烟鸡,攻毒后第4、7、10、14、21、28、35天采集外周血并分离外周血淋巴细胞,利用实时定量PCR技术对外周血淋巴细胞中IL-18转录水平进行相对定量分析。结果表明,普通霞烟鸡外周血淋巴细胞中具有相对高水平的IL-18mRNA,而马立克氏病抗性霞烟鸡外周血淋巴细胞中则具有相对低水平的IL-18mRNA。结果显示,鸡IL-18基因的表达模式与马立克氏病(MD)抗性遗传性状可能相关。     

Kinetics of phytohemagglutinin response in chickens infected with various strains of Marek's disease virus

Phytohemagglutinin (PHA) response of whole blood lymphocytes from white leghorn inbred line 7(2) chickens infected with various strains of Marek's disease virus (MDV) was monitored sequentially for 6 weeks postinfection. A significant difference between JM and GA strains was shown. A two-phase depression in the PHA response was observed in chickens infected with the JM strain. Early depression occurred 1 week postinfection and was followed by recovery a week later. The second depression occurred at 4 weeks postinfection and lasted until the end of the experiment. The GA strain-infected group, on the other hand, began to show depression 4 weeks postinfection, and most chickens died within a short time thereafter. PHA response of chickens infected with strain Md11/75C, attenuated in cell culture from highly virulent strain Md11, was almost the same as that of control chickens.     

Coinfection of specific-pathogen-free chickens with Marek's disease virus (MDV) and chicken infectious anemia virus: effect of MDV pathotype

Both Marek's disease virus (MDV) and chicken infectious anemia virus (CIAV) infections are prevalent in chickens throughout the world. In the past decade, MDV strains with increased virulence (very virulent plus MDV pathotype [vv+MDV]) have been isolated. The purpose of this experiment was to determine the effects of coinfection of chickens with CIAV and a vv+MDV isolate. Specific-pathogen-free chickens were inoculated at 1 day posthatch with RB1B (very virulent MDV pathotype [vvMDV]) only, 584A (vv+MDV) only, CIAV only, RB1B + CIAV, 584A + CIAV, or nothing. Samples of spleen, thymus, and bursa of Fabricius were collected at 4, 7, 10, and 13 days postinoculation (DPI). Thymic and bursal atrophy at 13 DPI and final mortality at 30 DPI were significantly greater in chickens inoculated with 584A with or without added CIAV, or with RB1B plus CIAV, compared with birds inoculated with RB1B alone. Both amounts of virus reisolated and levels of virus detected by quantitative-competitive polymerase chain reaction were greater at 4 DPI in 584A inoculates compared with RB1B inoculates. To monitor the early cytolytic infection, northern analysis was done with a probe for the MDV immediate early gene ICP4 (infected cell protein 4). In the absence of CIAV, ICP4 expression was more apparent in chickens inoculated with 584A than in those inoculated with RB1B. CIAV coinfection increased ICP4 expression in the spleens of chickens infected with RB1B. These results indicated that inoculation of chickens with the 584A isolate caused a more robust early cytolytic infection compared with inoculation with RB1B alone and support the classification of 584A as a vv+MDV strain. Coinfection with CIAV exacerbated vvMDV strain RB1B infection. The extent of this exacerbation was less evident when birds were coinfected with 584A and CIAV.     

On the specificity of the immunoglobulin G produced by chickens experimentally infected with Marek's disease virus

In chickens experimentally infected with Marek's Disease virus (MDV) an increased amount of immunoglobulin G is produced. Using a technique of quantitative crossed immunoelectrophoresis it has been shown, that 70% of this immunoglobulin G is non-specific. Only 18% could be absorbed with MDV strain CPRL VII-infected chicken kidney cells, and only 5% with MDV-induced lymphoblastoid cells of the MDCC-MSB1 cell line. It is hypothesized that the production the unspecific immunoglobulin G is caused by a polyclonal stimulation of B-cells.     

Marek's disease in field chickens: correlation between incidence of Marek's disease and nuclear-inclusion formation in the feather-follicle epithelium

The present study confirmed that Marek's disease (MD)-associated nuclear-inclusion (NI) formation in the feather-follicle epithelium (FFE) is related to mortality from MD; it also presented useful data on the epidemiology of MD in HVT-vaccinated field chickens. Incidence of NI formation in the FFE of chickens on six rearing farms varied greatly by age and flock, but most of the field flocks showed biphasic peaks of incidence of NI in chickens consisting of a small peak at an early age (usually at 2-4 weeks of age) and a large peak between 13 and 16 weeks of age. MD tended to occur in chickens over 20 weeks old, and almost all MD-affected chickens showed NI formation persistently in the FFE, usually between 13 and 20 weeks of age. Chickens that were healthy at the end of observation showed either transient NI formation, usually between 13 and 16 weeks of age, or no detectable NI formation. Incidence of NI formation in the FFE of chickens at 19-20 weeks of age was related to mortality from MD: chickens with NI formation had a MD mortality rate of 66.7%, whereas chickens without NI formation had MD mortality of only 0.8%.