Methyltransferase-defective avian metapneumovirus vaccines provide complete protection against challenge with the homologous Colorado strain and the heterologous Minnesota strain.

UNLABELLED: Avian metapneumovirus (aMPV), also known as avian pneumovirus or turkey rhinotracheitis virus, is the causative agent of turkey rhinotracheitis and is associated with swollen head syndrome in chickens. Since its discovery in the 1970s, aMPV has been recognized as an economically important pathogen in the poultry industry worldwide. The conserved region VI (CR VI) of the large (L) polymerase proteins of paramyxoviruses catalyzes methyltransferase (MTase) activities that typically methylate viral mRNAs at guanine N-7 (G-N-7) and ribose 2'-O positions. In this study, we generated a panel of recombinant aMPV (raMPV) Colorado strains carrying mutations in the S-adenosyl methionine (SAM) binding site in the CR VI of L protein. These recombinant viruses were specifically defective in ribose 2'-O, but not G-N-7 methylation and were genetically stable and highly attenuated in cell culture and viral replication in the upper and lower respiratory tracts of specific-pathogen-free (SPF) young turkeys. Importantly, turkeys vaccinated with these MTase-defective raMPVs triggered a high level of neutralizing antibody and were completely protected from challenge with homologous aMPV Colorado strain and heterologous aMPV Minnesota strain. Collectively, our results indicate (i) that aMPV lacking 2'-O methylation is highly attenuated in vitro and in vivo and (ii) that inhibition of mRNA cap MTase can serve as a novel target to rationally design live attenuated vaccines for aMPV and perhaps other paramyxoviruses. IMPORTANCE: Paramyxoviruses include many economically and agriculturally important viruses such as avian metapneumovirus (aMPV), and Newcastle disease virus (NDV), human pathogens such as human respiratory syncytial virus, human metapneumovirus, human parainfluenza virus type 3, and measles virus, and highly lethal emerging pathogens such as Nipah virus and Hendra virus. For many of them, there is no effective vaccine or antiviral drug. These viruses share common strategies for viral gene expression and replication. During transcription, paramyxoviruses produce capped, methylated, and polyadenylated mRNAs. Using aMPV as a model, we found that viral ribose 2'-O methyltransferase (MTase) is a novel approach to rationally attenuate the virus for vaccine purpose. Recombinant aMPV (raMPV) lacking 2'-O MTase were not only highly attenuated in turkeys but also provided complete protection against the challenge of homologous and heterologous aMPV strains. This novel approach can be applicable to other animal and human paramyxoviruses for rationally designing live attenuated vaccines.

Prevalence of parvoviruses in commercial turkey flocks.

Turkey parvovirus belongs to the family Parvoviridae, subfamily Parvovirinae, Genus parvovirus. Since the initial report on turkey parvovirus in the United States appeared in 1983, there had been no further reports of parvovirus in turkeys until 2008. The aims of our study were to determine the prevalence of parvovirus in commercial turkey flocks using PCR; to determine their genetic relationship to previous strains identified in North America and Europe; and to test samples for enteric viruses by transmission electron microscopy (TEM). A total of 169 fecal samples collected from 42 turkey farms in four different states within the United States between 2000 and 2010 were examined. We found that the most frequently detected viruses by TEM were small round viruses, accounting for 52% of the examined samples; however, the PCR detected parvoviruses in 71% of the samples. The phylogenetic analysis of partial nonstructural gene sequences showed a certain degree of variability among the turkey samples tested in the study. Moreover, there was a clear dichotomy in the phylogenetic tree between chicken and turkey samples, with the exception of one turkey isolate from 2000, which clustered together with the chicken group.

Pullorum Disease: Evolution of the Eradication Strategy.

The history of pullorum disease is closely intertwined with the history of avian health research and that of the poultry industry. The seriousness of the disease galvanized the attention and brought together, for the first time, the pioneers of poultry health research to work cooperatively on different aspects of the disease. Control of the disease made it possible for intensive poultry production to develop as the basis for the modern poultry industry. During the early 1900s, bacillary white diarrhea (BWD) was a devastating disease of young chickens threatening the developing poultry industry. Dr. Leo F. Rettger isolated and described the bacterial pathogen, Salmonella enterica serotype Pullorum, for the first time in 1900. BWD was renamed pullorum disease in 1929. In subsequent years, Rettger and coworkers were able to reproduce the disease and fulfill Koch's postulates. Rettger et al. also showed that Salmonella Pullorum was vertically transmitted, which was the first time that a pathogen was shown to be vertically transmitted. The development of serologic tests was of crucial importance because it led to the development of effective eradication methods to identify carrier birds and to exclude these birds from the breeder flocks. The negative impact of pullorum disease on the poultry industry ultimately was one of the major reasons that the National Poultry Improvement Plan (NPIP) was developed by scientists, the poultry industry, and the United States Department of Agriculture (USDA). Needless to say, the work of the pioneering researchers formed the basis for the control of the disease. The NPIP started in 1935, with 34 states participating in testing 4 million birds representing 58.2% of the birds hatched. The program rapidly expanded to 47 states by 1948 and tested more than 30 million birds. In 1967, all commercial chicken hatcheries participating in the NPIP were 100% free of pullorum and typhoid disease caused by Salmonella enterica serotype Gallinarum. This historical overview of pullorum disease describes in some detail the progress made, especially during the early years, toward controlling this disease using methodologies that were often very basic but nonetheless effective. One has to admire the ingenuity and persistence of the early researchers leading to their achievements considering the research tools that were available at the time.

Artículo histórico­Pulorosis: Evolución de las estrategias de erradicación La historia de la pulorosis está estrechamente relacionada con la historia de la investigación en salud aviar y de la industria avícola. La severidad de la enfermedad despertó la atención y reunió, por primera vez a los pioneros de la investigación en salud avícola para trabajar de manera cooperativa en diferentes aspectos de la enfermedad. El control de la enfermedad hizo posible que la producción avícola intensiva se desarrollara como base de la industria avícola moderna. A principios de la década de los 1900, la diarrea blanca bacilar (con las siglas en inglés BWD) era una enfermedad devastadora de pollos jóvenes que amenazaba la industria avícola en desarrollo. El Dr. Leo F. Rettger aisló y describió el patógeno bacteriano, Salmonella enterica serotipo Pullorum, por primera vez en 1900. La diarrea blanca bacilar pasó a llamarse pulorosis (pullorum disease) en 1929. En los años siguientes, Rettger y sus colaboradores pudieron reproducir la enfermedad y cumplir los postulados de Koch. Rettger y col. también mostraron que Salmonella Pullorum se transmitía verticalmente, y fue la primera vez que se demostró que un patógeno se transmitía verticalmente. El desarrollo de pruebas serológicas fue de crucial importancia porque condujo al desarrollo de métodos de erradicación efectivos para identificar aves portadoras y eliminar a estas aves de las parvadas reproductoras. El impacto negativo de la pulorosis en la industria avícola fue, en última instancia, una de las principales razones por las que los científicos, la industria avícola y el Departamento de Agricultura de los Estados Unidos (USDA) desarrollaron el Plan Nacional de Mejoramiento Avícola (NPIP). Es importante decir que el trabajo de los investigadores pioneros formó la base para el control de la enfermedad. El Plan Nacional de Mejoramiento Avícola comenzó en año 1935, con 34 estados participando en el análisis de 4 millones de aves que representaban el 58.2% de las aves producidas. El programa se expandió rápidamente a 47 estados en 1948 y evaluó a más de 30 millones de aves. En 1967, todas las plantas incubadoras de pollos comerciales que participaban en el Plan Nacional de Mejoramiento Avícola estaban 100% libres de pulorosis y tifoidea aviar causada por Salmonella enterica serotipo Gallinarum. Esta reseña histórica de la pulorosis describe con cierto detalle el progreso realizado, especialmente durante los primeros años, hacia el control de esta enfermedad utilizando metodologías que a menudo eran muy básicas no obstante efectivas. Es admirable el ingenio y la persistencia de los primeros investigadores que los llevaron a sus logros considerando las herramientas de investigación que estaban disponibles en ese momento.

Pathobiological characterization of low-pathogenicity H5 avian influenza viruses of diverse origins in chickens, ducks and turkeys.

We undertook one of the most comprehensive studies on the replication and intraspecies transmission characteristics of low-pathogenicity avian influenza viruses in ducks, chickens and turkeys. Our results indicated that most of these isolates could replicate and be transmitted in poultry without inducing clinical disease. However, differences in transmission to contact control birds were noted, emphasizing the importance of having contact control cage mates in biological characterization experiments. Ducks supported the replication of viruses of wild aquatic bird origin in their respiratory and digestive tracts equally well. The viruses from wild aquatic birds were not effectively transmitted among chickens. In contrast, the wild-bird isolates and viruses of domestic bird origin from live-bird markets and commercial poultry operations replicated and were transmitted more efficiently in turkeys than in chickens or ducks. We also found a lower minimal infectious dose requirement for infection of turkeys compared to chickens and ducks. Our data support an important role of turkeys as being more susceptible hosts for avian influenza viruses than domestic ducks and chickens. These results highlight the role of turkeys as intermediate or bridging hosts in the transmission of influenza viruses from wild birds to land-based domestic poultry or among different land-based bird species.

Detection of influenza A viruses in eggs laid by infected turkeys.

Studies are limited on evaluating the potential of influenza viruses for egg-borne dissemination. In our previous studies, experimental infection of breeder turkeys with A/turkey/Ohio/313053/04 resulted in drastic declines in egg production, and we confirmed high levels of virus replication and an abundant distribution of avian-specific alpha2,3 sialic acid-gal receptors in the oviduct of these turkeys. In the present study, following experimental inoculation of A/turkey/Ohio/313053/04 in breeder turkeys, we detected these viruses in the albumin of eggs using real-time RT-PCR (RRT-PCR) and virus isolation in embryonated chicken eggs. Swabs from egg shells were also found positive by RRT-PCR. This is the first report of the detection of low pathogenic influenza viruses from internal egg contents following experimental infection. The possibility of hatchery contamination by egg-borne influenza viruses, and the spread of virus during movement of contaminated cracked eggs and egg flats, pose concerns regarding viral dissemination of influenza.

The high susceptibility of turkeys to influenza viruses of different origins implies their importance as potential intermediate hosts.

Several previous reports and our studies show that waterfowl-origin influenza viruses can be more easily transmitted to domestic turkeys than chickens. Similarly, studies indicate turkeys to be better hosts for low pathogenic avian influenza viruses isolated from commercial poultry operations and live bird markets in comparison to chickens. Low 50% infectious-dose titers of wild bird as well as poultry-adapted viruses for turkeys further suggest that turkeys can be easily infected following a low-dose exposure. Also, interspecies transmission of swine influenza viruses to turkeys occurs frequently. These findings suggest the role of turkeys as suitable intermediate hosts that can be easily infected with influenza viruses of different origins and that turkeys can act as source of infection for other land-based poultry or even mammals.

Development of differential RT-PCR assays and molecular characterization of the complete VP1 gene of five strains of very virulent infectious bursal disease virus.

Several antigenic and pathogenic subtypes of infectious bursal disease virus (IBDV) have been identified worldwide. Simple and quick differential diagnostic assays are vital for implementing control and prevention strategies for infectious bursal disease (IBD). Currently reverse transcriptase-polymerase chain reaction/restriction fragment length polymorphism analysis (RT-PCR/RFLP) and real-time RT-PCR detection have been used. However, the RT-PCR/RFLP analysis is time consuming and not feasible for assaying large numbers of samples, and the real-time PCR is expensive. The reliable indicator for very virulent (vv) IBDV (vvIBDV) remains in vivo pathogenicity testing because of the lack of a virulence marker. In this study simple RT-PCR assays were developed for differentiating various types of IBDV. Two sets of primers specific for serotype 2 and vvIBDV were designed based on the sequences of segment A and B of IBDVs. Initially a total of 25 previously characterized virus strains including 11 serotype 1 classics, 4 serotype 1 variants, and 5 serotype 1 vv and 5 serotype 2 strains were used to validate the differential RT-PCR assays. The results indicated that primer set 1 specifically amplified a 415 bp RT-PCR product for the serotype 2 viruses, and primer set 2 specifically amplified a 715 RT-PCR product for vvIBDV except for two vv Taiwan strains. To further confirm the specificity of primer set 2 for the vvIBDVs, 20 field samples suspected to be vvIBDVs from different geographic locations around the world were tested. All but one suspected vv Korean strain (91108) tested positive by this primer set. To understand the molecular basis for the failure to detect these viruses with primer set 2, a complete VP1 gene of two vv Taiwan (PT and IL) strains along with two vv Turkey (OA and OE) and one vv Holland (Hol) strain was sequenced and phylogenetically analyzed with 11 other strains retrieved from GenBank. The results showed that PT and IL were closely related to the classic strain IM with nucleotide (nt) similarities of 98.6% and 97.7%, respectively. The two Taiwan strains clustered together with the classic and variant IBDVs in the unrooted neighbor-joining phylogenetic tree, indicating independent evolution of these strains from the rest of the vv isolates. The RT-PCR assays developed in this study could differentiate vvIBDVs from classic strains and serotype 1 from serotype 2 IBDVs with a high degree of sensitivity and specificity and are fast, simple, and inexpensive.

Detection of antibodies against serotypes 1 and 2 infectious bursal disease virus by commercial ELISA kits.

Two distinct serotypes of infectious bursal disease virus (IBDV) are recognized in chicken and turkey flocks in the United States. Serologic testing of chicken flocks for serotype 1 viruses is routinely performed to monitor disease status and vaccination. Earlier studies indicated that enzyme-linked immunosorbent assay (ELISA) test detects antibodies to both serotypes of the virus, while the virus neutralization (VN) test is serotype specific. It is useful to evaluate currently available commercial ELISA kits for their ability to differentiate between antibodies elicited by the two serotypes. Three trials were performed in which chickens were orally inoculated with either a high or a low dose of serotype 1 STC or serotype 2 OH strains of IBDV. Sera collected at 0, 7, 14, and 21 days from these chickens and antisera procured from naturally infected broiler (n=20) and layer (n=30) flocks were tested with five different commercial ELISA kits and by VN. All ELISA kits detected different levels of antibodies elicited against serotype 1 of the virus and moderate and high levels of antibodies against serotype 2 virus. A correlation existed between the ELISA and the VN titers of experimentally infected chickens. All serum samples tested from the commercial layer flocks and 65% of the broiler flocks had antibodies against the OH strain. However, no correlation between the VN titers and ELISA titers was observed for the commercial broilers and layers sera by the majority of the kits. The results indicated that currently available commercial ELISA kits detect antibodies elicited by the two serotypes of IBDV. Hence, the prevalence of serotype 2 antibodies in the flocks should be considered while determining antibody profiles of the flocks against serotype 1 viruses.

Pathogenicity of turkey astroviruses in turkey embryos and poults.

The pathogenicity of turkey astrovirus 2001 (TAstV2001) and turkey astrovirus 1987 (TAstV1987) in specific-pathogen-free (SPF) turkey embryos and commercial poults was investigated. The virus shedding in poults was monitored using electron microscopy (EM) and reverse transcription-polymerase chain reaction (RT-PCR) during the 14-day experimental period. Both viruses caused enteritis and growth depression in SPF turkey embryos and poults. The TAstV2001 did not induce macroscopic or microscopic lesions in thymuses and bursas of embryos or poults. No macroscopic changes were observed in thymuses and bursas of embryos and poults inoculated with TAstV1987, and no statistically significant differences in bursa weight/ body weight ratios (P > 0.05) were detected. However, TAstV1987 infection resulted in microscopic lesions in bursas but not in thymuses of infected embryos and poults. Both TAstV2001 and TAstV1987 were shed during the whole 14-day experimental period as detected by EM and RT-PCR. These findings indicated that both TAstV1987 and TAstV2001 are etiologic agents of turkey enteritis. In addition, TAstV1987 might cause impairment of the immune system of infected poults. The pathogenicity of TAstV1987 is somewhat different from TAstV2001.

Development of a ssRNA internal control template reagent for a multiplex RT-PCR to detect turkey astroviruses.

Turkey astrovirus (TAstV), a positive sense single-stranded RNA (ssRNA) virus, is an important causative agent of enteritis of poults. The detection and diagnosis of astroviruses have been mainly dependent on electron microscopy (EM). However, EM is not very sensitive. Reverse transcription-polymerase chain reaction (RT-PCR) has high specificity and sensitivity. Thus, monoplex RT-PCR and multiplex RT-PCR for detection of TAstVs were developed in our laboratory. RT-PCR could be adversely affected by many factors, which would result in lowering the sensitivity of the reaction. To minimize this pitfall of RT-PCR, a ssRNA internal control (IC) template reagent containing two sets of primer sequences (SRV and AFCP) specific to the capsid region of TAstV genomes was developed, and applied to the multiplex RT-PCR. Sixty-four fecal specimens from 2-week-old poults with enteritis were tested using the multiplex RT-PCR with the IC serving as a co-amplification template. An overall test inhibition rate of 12.5% was found for the RT-PCR, which can be used to decrease the false-negative rate. The approach to the generation of the IC developed in this study is simple, convenient and productive, and can be used as a universal protocol to generate a ssRNA IC template reagent for RT-PCR.

Development of antigen-capture enzyme-linked immunosorbent assay and RT-PCR for detection of turkey astroviruses.

Turkey astrovirus (TAstV) is an important agent of poult enteritis. The diagnosis of astroviruses has been dependent mainly on electron microscopy (EM) or immune EM (IEM). To develop other simple, rapid, and reliable diagnostic assays, two antigen-capture enzyme-linked immunosorbent assays (AC-ELISAs), polyclonal AC-ELISA and monoclonal AC-ELISA, were developed in this study. Monoplex and multiplex reverse transcription-polymerase chain reactions (RT-PCRs) were also developed using nondegenerate primer sets specific to the capsid region and degenerate primer pairs specific to the polymerase area of two TAstV. EM was included for comparison. Fecal or intestinal contents samples from naturally and experimentally infected poults with enteritis were examined using the developed assays. The polyclonal AC-ELISA had higher sensitivity and wider detection spectrum than the monoclonal AC-ELISA with group-specific monoclonal antibody (MAb), whereas the monoclonal AC-ELISA had very high specificity but lower sensitivity, which was estimated at 0.06 microg of viral proteins. Small round viruses (SRV) that could be astroviruses or other small viruses were detected in 34.4% of the samples examined by EM. The monoplex RT-PCR results amplified with primers SRV-1-3 and SRV-1-5 revealed that the positive rate of astroviruses was 45.3%, which was 10.9% higher than that of EM even if other SRVs were not excluded. Multiplex RT-PCR with SRV-1-3 and SRV-1-5 and AFCP-F1 and AFCP-R1 and the monoplex RT-PCR with degenerate primers verified that the positive rate of astroviruses was 59.4%, which was 25% higher than that of EM. Both RT-PCRs showed good specificity and wider detection spectrum compared with earlier published data.

Molecular characterization of the capsid gene of two serotypes of turkey astroviruses.

Astrovirus infections mainly cause acute gastroenteritis in children and young animals. Human astroviruses are well characterized antigenically and genetically. However, information on turkey astroviruses is limited. We isolated two astroviruses (TAstV1987 and TAstV2001) from turkeys and classified them as two different serotypes using a virus neutralization test. To elucidate the differences between these two isolates at the molecular level, further genetic characterization and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis were carried out. The sequences of the complete capsid protein gene of these two isolates were obtained by cloning and sequencing. The percentage nucleotide and predicted amino acid identities for these two sequences along with those of 16 other capsid protein gene sequences from human and animal astroviruses retrieved from GenBank were calculated using MegAlign. The results showed that TAstV1987 and TAstV2001 had 73.3% nucleotide and 82.8% amino acid identities, respectively. An unrooted Neighbor-joining phylogenetic tree of these sequences was generated using MEGA 3 software with 1000 bootstrap replicates. The results of evolutionary analysis showed that TAstV1987 was closely related genetically to another virus, designated TAstV-2, whereas TAstV2001 was not as close to TAstV-2 as TAstV1987. The analysis of the capsid proteins of the two viruses by SDS-PAGE revealed that they had different band patterns, indicating that their capsid proteins consisted of different viral proteins. The findings in this study revealed the molecular differences in the capsid protein gene of TAstV1987 and TAstV2001, which may provide the molecular basis of the antigenic differences between these two serotypes of turkey astroviruses.

Interference between mild and pathogenic strains of infectious bursal disease virus in chickens.

Infectious bursal disease virus is a contagious, immunosuppressive disease of young chickens that is controlled by vaccination. Cross-protection occurs between different strains of the virus as a result of shared neutralizing epitopes. However, interactions between two antigenically similar strains (a mild and a pathogenic) coinfecting the same host have not been investigated. Groups of specific-pathogen-free chickens were inoculated with a mild strain followed by a pathogenic strain at 0, 16, 24, or 48 hr postinoculation (PI) with a mild strain. Virus persistence and the predominant strain of the virus were determined by reverse transcriptase-polymerase chain reaction and restriction fragment length polymorphism analysis, respectively, in bursas at 2, 4, 8, 14, and 21 days PI with the pathogenic strain. Severity of infection was assessed by the bursa/body weight ratios and histopathologic lesion scores. The mild virus interfered with replication of the pathogenic virus. The greatest interference was observed when the pathogenic strain was inoculated 24 hr PI with the mild strain. The interference phenomenon observed might be due to competition for host receptor sites or production of cytokine(s). This interference phenomenon could have practical implications for vaccine usage and protection.

Isolation and characterization of H3N2 influenza A virus from turkeys.

Five 34-wk-old turkey breeder layer flocks in separate houses of 2550 birds each in a single farm in Ohio experienced a drop in egg production from late January to early February 2004. Tracheal swabs (n = 60), cloacal swabs (n = 50), and convalescent sera (n = 110) from the flocks were submitted to the laboratory for diagnostics. Virus isolation was attempted in specific-pathogen free embryonating chicken eggs and Vero and MDCK cells. Virus characterization was performed using agar gel immunodiffusion, the hemagglutination test, the hemagglutination inhibition test, the virus neutralization test, reverse transcription-polymerase chain reaction, sequencing, and phylogenetic analysis. A presumptive influenza virus was successfully propagated and isolated on the first passage in MDCK cells, but initially not in Vero cells or specific-pathogen free chicken embryos. After two passages in MDCK cells, it was possible to propagate the isolate in specific-pathogen free chicken embryos. Preliminary sequence analysis of the isolated virus confirmed that it was influenza A virus with almost 100% (235/236) identity with the matrix gene of a swine influenza A virus, A/Swine/Illinois/100084/01 (H1N2). However, it was not possible to subtype the virus using conventional serotyping methods. The results of genetic characterization of the isolated virus showed that it was the H3N2 subtype and was designated as A/Turkey/OH/313053/04 (H3N2). Phylogenetic analysis of the eight gene segments of the virus showed that A/Turkey/OH/313053/04 (H3N2) isolate was most closely related to the triple-reassortant H3N2 swine viruses [A/Swine/WI/14094/99 (H3N2)] that have been circulating among pigs in the United States since 1998, which contains gene segments from avian, swine, and human viruses. The A/Turkey/OH/313053/04 (H3N2) isolated from turkeys in this study was classified as a low pathogenic avian influenza A virus because it only caused a drop in egg production with minor other clinical signs and no mortality.

In-vitro antimicrobial susceptibility of Clostridium perfringens from commercial turkey and broiler chicken origin.

The minimum inhibitory concentrations (MIC) of eight antibiotics and two anticoccidial agents were determined for Clostridium perfringens strains isolated from 26 commercial broiler farms and 22 commercial turkey farms. Isolates were obtained from the intestines of birds on the farm or as the processing plant using standard culture and identification techniques. The microbroth dilution test was used to determine the MIC for each compound. Most isolates from chickens had MICs in the range of 2-16 mg/L for tilmicosin, tylosin and virginiamycin, whereas the MICs for avilamycin, avoparcin, monensin, narasin and penicillin were < or = 1 mg/L. Most strains from chickens had high MICs (> or = 64 mg/L) and appeared to be resistant to bacitracin and lincomycin. Most turkey isolates had MICs in the range of 2-16 mg/L for bacitracin, tilmicosin, tylosin and virginiamycin, with strains exhibiting MICs < or = 1 mg/L for avilamycin, avoparcin, monensin, narasin and penicillin. Several turkey isolates had MICs > or = 64 mg/L to lincomycin. No attempt was made to associate farm usage of a particular antibiotic to the antibiograms.

Immunosuppression induced by infectious bursal disease virus.

Immunosuppression caused by infectious bursal disease virus (IBDV) is of major interest because of the widespread occurrence of the infection in commercial chickens. Infection with IBDV at an early age significantly compromises the humoral and local immune responses of chickens. The cellular immune response is also compromised by apparently to a lesser extent and for a short period. The immunosuppression seems to be a result of direct effect (lysis) of B cells or their precursors. Other mechanisms of immunosuppression have been suggested, notably the development of suppressor cells.

Development of an antigen-capture enzyme-linked immunosorbent assay for detection of enterovirus in commercial turkeys.

Diagnosis of enterovirus infection in turkey poults is currently done using electron microscopy. We have developed an antigen-capture enzyme-linked immunosorbent assay (ELISA) for use in diagnosis of enterovirus infection in turkey intestinal contents. Intestinal contents from naturally and experimentally infected turkey poults were evaluated for the presence of astrovirus and enterovirus by immune electron microscopy (IEM). The results were compared with the findings obtained using the ELISA for the presence of enterovirus. The ELISA, using the samples from the naturally infected poults, had a sensitivity of 0.963 and a specificity of 0.879. The Kappa value of 0.834 for the naturally infected samples and 0.97 for the experimentally infected samples showed excellent agreement between the ELISA and IEM for detection of enterovirus. Our results indicate that the ELISA is a rapid, sensitive, and specific method for diagnosis of turkey enterovirus infection.