Avian leukosis virus subgroup J infection profiles in broiler breeder chickens: association with virus transmission to progeny

Profiles of infection with avian leukosis virus subgroup J (ALV-J) and factors that predict virus transmission to progeny were studied. Eggs from an infected broiler breeder flock were hatched at the laboratory. The flock was reared in a floor pen, transferred to laying cages at 22 wk, and inseminated to produce fertile eggs. A cohort of 139 chickens was tested at frequent intervals over a 62-wk period for virus, viral antigens, or antibodies in plasma, cloacal swabs, egg albumen, and embryos. Virus was detected in 7% of chicks at hatch but spread rapidly so that virtually all chicks became infected between 2 and 8 wk of age. Mortality due to myeloid leukosis and related tumors was 22%. Over 40% of the chicks developed persistent infections, whereas the remainder experienced transient infections. Five types of infection profiles were recognized. Novel responses included hens that were positive for virus intermittently or started late in life to shed viral antigens into the cloaca. ALV-J was isolated from 6% of 1036 embryos evaluated between 26 and 62 wk. However, over 90% of the virus-positive embryos were produced between 29 and 34 wk of age. Of 80 hens that produced embryos, 21 produced at least one infected embryo and were identified as transmitters. All but one transmitter hen would have been detected by a combination of viremia, cloacal swab, and albumen tests conducted between 18 and 26 wk. However, virus was transmitted to embryos from hens that were not persistently viremic or that rarely shed viral group-specific antigen into the albumen of their eggs. Intermittent patterns of both antigen shedding and virus transmission to embryos were observed in some hens. These results validate current screening procedures to identify potential transmitter hens and provide some suggestions for improvement but also show that identification of all transmitter hens by such procedures is unlikely. Thus, eradication programs based solely on dam testing may be less effective than those where dam testing is combined with procedures to mitigate early horizontal transmission in progeny chicks.     

Avian leukosis virus and selection on oviposition interval in Australorp lines.

1. Shedding of group-specific antigen of avian leukosis virus (ALV) into egg albumen was determined in three Australorp lines: AS line that had been selected primarily for short oviposition interval, ASS line that had been derived from the AS line and developed as a commercial dam line for egg laying, and AC line that had been kept as a randombred control. 2. The proportion of shedders was 13.1 to 16.7% in the AS line in 1984-88, 16.3% in the ASS line in 1984 (before culling of the shedders), and 6.1% and 6.6% in the AC line in 1984 and 1988, respectively. 3. In the AS line, shedders were 1.8% lower in rate of lay to 300 d of age, 1.3 g lower in average egg weight at 34 weeks of age, 5.6% lower in hatchability of fertile eggs and 0.24 h shorter in oviposition interval than non-shedders. In the ASS line (1984 only), the differences between shedders and non-shedders were in the same direction, but in magnitude greater for rate of lay and smaller for oviposition interval. 4. The shedders were favoured by the artificial selection because of their shorter oviposition interval and this appeared to be responsible for the higher levels of ALV shedding in the selection lines.     

Avian leukosis virus (ALV) infection, shedding, and tumors in maternal ALV antibody-positive and -negative chickens exposed to virus at hatching

Chickens highly susceptible to avian leukosis virus (ALV) infection and tumors, with and without ALV subgroup A maternal antibody (MAB), were infected with a field strain of ALV subgroup A at hatching. Viremia, antibody development, cloacal and albumen shedding, and tumors in chickens with MAB (MAB+) were compared with those in chickens lacking MAB (MAB-). At 18 weeks of age, the incidence of viremia was significantly lower in MAB+ chickens than in MAB- chickens; further, MAB significantly reduced the proportion of tolerantly infected (viremic antibody-negative) chickens. Cloacal shedding of ALV at 22 weeks of age and shedding of ALV group-specific (gs) antigen in albumen of eggs from all laying hens at 30-32 weeks of age were significantly lower in MAB+ hens than in MAB- hens. The incidence of ALV-induced tumors was lower in MAB+ chickens than in MAB- chickens, significantly so in one of three trials conducted. These results suggest that MAB may influence the development of viremia, antibody, and shedding of ALV following massive exposure to virus at hatching.     

Replication-competent endogenous avian leukosis virus in commercial lines of meat chickens

Group-specific (gs)-antigen-positive egg albumen in seven commercial lines of meat chickens was found to result from the presence of endogenous avian leukosis virus (ALV); these lines had resisted selection attempts to reduce the shedding rate. In two meat lines, exogenous as well as endogenous ALV contributed to the gs-antigen shedding. All hens that produced gs-antigen-positive albumen transmitted endogenous ALV to a high proportion of their embryos (20 to 100%). Hens shedding gs-antigen to albumen were negative for endogenous ALV in vaginal swabs and had no detectable antibody to subgroup E virus. Chickens hatched from these dams were negative for endogenous ALV in meconia but were viremic at 2 weeks of age. Replication-competent endogenous ALV was almost uniformly expressed in embryos of hens from nine meat lines that were negative for gs-antigen in albumen. Shedding of gs-antigen to albumen was not related to the level of endogenous ALV expression. Embryos from five meat lines tested were resistant to infection with ALV of subgroup E. The level of endogenous gs-antigen in albumen was consistently lower than the level of exogenous gs-antigen.     

Response of white leghorn chickens of various genetic lines to infection with avian leukosis virus subgroup J

In Experiment 1, chickens from various white leghorn experimental lines were inoculated with strain ADOL-Hcl of subgroup J avian leukosis virus (ALV-J) either as embryos or at 1 day of age. At various ages, chickens were tested for ALV-J induced viremia, antibody, and packed cell volume (PCV). Also, at 4 and 10 wk of age, bursal tissues were examined for avian leukosis virus (ALV)-induced preneoplastic lesions with the methyl green-pyronine (MGP) stain. In Experiment 2, chickens harboring or lacking endogenous virus 21 (EV21) were inoculated with strain ADOL-Hcl of ALV-J at hatch. All embryo-inoculated chickens in Experiment 1 tested positive for ALV-J and lacked antibody throughout the experimental period of 30 wk and were considered viremic tolerant, regardless of line of chickens. By 10 wk of age, the incidence of ALV-J viremia in chickens inoculated with virus at hatch varied from 0 (line 0 chickens) to 97% (line 1515); no influence of ALV-J infection was noted on PCV. Results from microscopic examination of MGP-stained bursal tissues indicate that ALV-J can induce typical ALV-induced transformation in bursal follicles of white leghorn chickens. Lymphoid leukosis and hemangiomas were the most common ALV-J-induced tumors noted in chickens in Experiment 1. At termination of Experiment 2 (31 wk of age), 54% of chickens harboring EV21 were viremic tolerant compared with 5% of chickens lacking EV21 after inoculation with ALV-J at hatch. The data indicate that genetic differences among lines of white leghorn chickens, including the presence or absence of EV21, can influence response of chickens to infection with ALV-J.     

Detection of subgroup J avian leukosis virus infection in Australian meat-type chickens

OBJECTIVE: To determine the extent of avian leukosis virus subgroup J (ALV-J) infection in Australian broiler breeder flocks, using virus isolation and molecular biological detection. Any resultant ALV-J viral isolates to be characterised by neutralisation cross testing in order to determine antigenic relationships to overseas isolates of ALV-J. STUDY DESIGN: Samples of blood, feather pulp, albumen and tumours were obtained from broiler breeder flocks which represented four genetic strains of meat chickens being grown in Victoria, South Australia, NSW and Queensland. Dead and ailing birds were necropsied on farm and samples were collected for microscopic and virological examinations. Virus isolation was carried out in C/O and DF-1 CEF cultures and ALV group specific antigen was detected in culture lysates using AC-ELISA. Micro-neutralisation assay was used for antigenic characterisation of selected isolates. Genomic DNA was isolated from cultured cells, tumours and feather pulp. ALV-J envelope sequences were amplified by PCR using specific ALV-J primers while antibodies against ALV-J were detected by ELISA. RESULTS: A total of 62 ALV-J isolates were recovered and confirmed by PCR from 15 (31.3%) of 48 breeder flocks tested. Antibody to ALV-J was detected in 20 (47.6%) of the 42 flocks tested. Characteristic lesions of myeloid leukosis caused by ALV-J were found in affected flocks. The gross pathological lesions were characterised by skeletal myelocytomas located on the inner sternum and ribs, neoplastic enlargement of the liver, and in some cases gross tumour involvement of the spleen, kidney, trachea, skeletal muscles, bone marrow, skin and gonads. Microscopically, the tumours consisted of immature granulated myelocytes, and were present as focal or diffuse infiltrations in the affected organs. Virus micro-neutralisation assays demonstrated antigenic variation among Australian isolates and to overseas strains of ALV-J. CONCLUSION: ALV-J infection was prevalent in Australian broiler breeder flocks during 2001 to 2003. Australian isolates of ALV-J show a degree of antigenic variation when compared to overseas isolates.     

Congenital transmission of avian leukosis virus in the absence of detectable shedding of group specific antigen

Congenital transmission of avian leukosis virus (ALV) in the absence of detectable amounts of group specific (gs) antigen in egg albumen was found to occur in one commercial and one specific pathogen-free (SPF) flock. The prevalence of congenitally transmitting hens which did not excrete gs antigen was particularly high in a commercial flock where 26/27 hens transmitted ALV. Some of the ALV-transmitting hens in the commercial flock had virus in vaginal swabs thus enabling infection to be detected. The reasons for such a high proportion of congenitally transmitting hens which did not shed detectable amounts of gs antigen in the commercial flock was not determined. In the SPF flock, 2/15 hens congenitally transmitted ALV although virus could not be detected in vaginal swabs, whole blood or egg albumen and antibodies to subgroups A or B were not present. This form of ALV-infection persisted in two successive generations. These results indicate the necessity of testing for infectious ALV in embryos, in order to ascertain that a flock is genuinely free of ALV.     

Myxosarcomas associated with avian leukosis virus subgroup A infection in fancy breed chickens

Formalin-fixed suspect tumors were submitted to the Poultry Diagnostic and Research Center at the University of Georgia (Athens, GA) for diagnosis. Samples were from fancy breed chickens with a history of increased tumor prevalence in both hens and roosters. Microscopically, in all the samples, there were neoplastic proliferations of spindle-shaped cells. The matrix surrounding tumor cells stained positively with Alcian blue at pH 2.5, but neoplastic cells did not stain with periodic acid-Schiff. Immunohistochemistry stains were positive for vimentin and neuron-specific enolase and negative for desmin, smooth muscle actin, and S-100 protein. Tumors were determined to be myxosarcomas. All samples were positive for PCR targeting the gp85 avian leukosis virus (ALV) envelope protein. However, analysis of the predicted amino acid sequences in the envelope gene from three separate samples showed high similarity between them and to ALV subgroup A.     

Association of the major histocompatibility complex with avian leukosis virus infection in chickens.

1. Association of the B blood group, the major histocompatibility complex (MHC) in chickens, with avian leukosis virus (ALV) infection shown by shedding of group-specific (gs) antigen was studied in an Australorp line selected for short oviposition interval to improve egg production. Three haplotypes (B8a, B9a and B21) were segregating in this line at frequencies of 66.7, 15.6 and 17.8%, respectively, averaged over three generations. 2. The relative risk (odds ratio) of a hen becoming a gs-antigen shedder was calculated for progenies of the dams shedding gs-antigen and those of non-shedding dams separately and pooled over three generations. In the progenies of shedding dams, the relative risk was not significantly different from 1.0 for the three haplotypes. In contrast, in the progenies of non-shedding dams, the relative risk was 0.67, 0.48 and 2.53 for B8a, B9a and B21, respectively, with the last two ratios being significantly different from 1.0. 3. The average effect of haplotype substitution on probability of shedding was estimated from a linear logistic model. The estimates (relative to zero for B8a) for B9a and B21, respectively, were -0.26 and 0.03 among the progenies of shedding dams, and -0.16 and 0.87 among the progenies of non-shedding dams. The last estimate only was highly significant. 4. These results suggest that the three haplotypes were similar in susceptibility to congenital infection through hatching eggs, but differed in susceptibility to post-hatching infection from other infected birds.     

Sporadic congenital transmission of avian leukosis virus in hens discharging the virus into the oviducts.

The efficacy of the albumen test for infectious avian leukosis virus (ALV) was examined in detecting congenitally transmitting hens. Seventy-three White Leghorn non-viremic hens with antibody to ALV were used. Eleven of the hens shed infectious ALV into their egg albumen, whereas only 7 of the 11 ALV-positive hens shed ALV antigens. The egg albumen test for infectious ALV was shown to be more effective in detecting the congenitally transmitting hens than that for ALV antigens. Then, twenty of the 62 hens which shed no infectious ALV into the albumen were studied for transmission of ALV to their embryos and for discharging ALV into the oviduct and vagina. Six of the 50 embryos from 4 hens were found to be infected with ALV but all of the 227 embryos from remaining 16 hens were free from the infection. Discharge of the virus into the oviduct and vagina was found both in the 4 transmitting hens and in 6 of the 16 non-transmitting hens. These results suggest that the hens discharging ALV into the oviduct, even though they do not shed ALV into egg albumen, may transmit the virus sporadically to their embryos.     

Response of white leghorn chickens to infection with avian leukosis virus subgroup J and infectious bursal disease virus

The effects of viral-induced immunosuppression on the infectious status (viremia and antibody) and shedding of avian leukosis virus (ALV) were studied. Experimental white leghorn chickens were inoculated with ALV subgroup J (ALV-J) and infectious bursal disease virus (IBDV) at day of hatch with the ALV-J ADOL prototype strain Hcl, the Lukert strain of IBDV, or both. Appropriate groups were exposed a second time with the Lukert strain at 2 wk of age. Serum samples were collected at 2 and 4 wk of age for IBDV antibody detection. Samples for ALV-J viremia, antibody detection, and cloacal shedding were collected at 4, 10, 18, and 30 wk of age. The experiment was terminated at 30 wk of age, and birds were necropsied and examined grossly for tumor development. Neoplasias detected included hemangiomas, bile duct carcinoma, and anaplastic sarcoma of the nerve. Control birds and IBDV-infected birds were negative for ALV-J-induced viremia, antibodies, and cloacal shedding throughout experiment. By 10 wk, ALV-J-infected groups began to develop antibodies to ALV-J. However, at 18 wk the incidence of virus isolation increased in both groups, with a simultaneous decrease in antibody levels. At 30 wk, 97% of birds in the ALV-J group were virus positive and 41% were antibody positive. In the ALV-J/IDBV group, 96% of the birds were virus positive at 30 wk, and 27% had antibodies to ALV-J. In this study, infection with a mild classic strain of IBDV did not influence ALV-J infection or antibody production.     

Immune response to avian leukosis virus infection in chickens: Sequential expression of serum immunoglobulins and viral antibodies

Chickens of a 15I₅ × 7₂ cross that produces endogenous Rous associated virus (RAV-0) were infected with subgroup A lymphoid leukosis virus (RAV-1). Within 3 weeks, before RAV-1 neutralizing antibodies were detected, significantly higher levels of serum immunoglobulin G (IgG) were found in infected birds than in uninoculated hatchmates. Immunoglobulin M was significantly elevated only during the late leukotic state. Although most of the inoculated birds tested had RAV-1 neutralizing antibodies, no correlation was found between IgG levels and antibody titers. Tolerance to endogenous virus (RAV-0) and viral group-specific antigen was apparently abrogated by RAV-1 inoculation because significantly higher percentages of iodinated envelope glycoprotein (gpE) of RAV-0 and a viral structural antigen of mol. wt 19,000 daltons (p 19) were precipitated by sera from inoculated birds than from control birds.     

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.     

禽白血病病毒跨膜蛋白在膜融合及免疫抑制中的作用

禽白血病病毒（Avian leukosis virus,ALV）入侵宿主细胞的关键是病毒裳膜与宿主细胞膜的融合,AI。V在此过程中采用病毒一宿主细胞膜融合机制,这一机制的关键是病毒与细胞受体结合后跨膜蛋白一系列构象的变化。此外,ALV入侵宿主细胞后会引起严重的免疫抑制,继而引发肿瘤的产生。在对一些与ALV有相同感染机制的病毒的研究中发现,引起免疫抑制的关键区也是跨膜蛋白。因此进一步在分子水平上深入研究跨膜蛋白有助于正确认识病毒侵染的本质,做到更为有效的预防与治疗ALV感染。