Sialodacryoadenitis in the rat: effects of immunosuppression on the course of the disease

Eight-week-old outbred male and female Crl:CD(SD)BR rats were treated with prednisolone (PR) or cyclophosphamide (CY) and were inoculated intranasally with sialodacryoadenitis (SDA) virus. The course of the disease was compared with nonimmunosuppressed, SDA virus-inoculated rats of the same stock. Criteria used to compare SDA in the 3 groups, included histologic changes in salivary and lacrimal glands, immunofluorescent microscopy of paraffin-embedded tissues, serum amylase levels, and antibody response. Based on these criteria, there was little detectable difference in the course and intensity of SDA in PR-treated and nonimmunosuppressed rats. In CY-treated rats, there was a delay in the onset of SDA, in the appearance of inflammatory cells in affected glands, and in the reparative process in the salivary and lacrimal glands. Viral antigen persisted longer in CY-treated rats than in PR-treated and nonimmunosuppressed rats. Antibody to SDA virus was not detected in CY-treated rats. The efficacy of immunosuppression by PR and CY was confirmed by the sheep erythrocyte agglutination procedure performed in selected rats. Male and female rats of the same strain were immunosuppressed beginning 4 weeks after inoculation with SDA virus to produce recrudescence of the disease. Histologic examination of salivary and lacrimal glands, immunofluorescent microscopy, serum amylase values, and viral isolation studies did not reveal evidence of reactivation of a persistent viral infection or viral shedding. Based on these studies, there is no evidence that SDA virus may persist as an inapparent infection after recovery from the disease.     

Depletion of salivary gland epidermal growth factor by sialodacryoadenitis virus infection in the Wistar rat

Male and female Wistar rats 2 to 15 months of age were inoculated intranasally with sialoda-cryoadenitis (SDA) virus and killed at 8 to 21 days post-inoculation (PI). Submandibular glands were evaluated by light and electron microscopy, and levels of salivary gland epidermal growth factor (EGF) were quantitated by cytochemistry and competitive radioreceptor assay. Apical granules in the epithelial cells of the granular convoluted tubules (GCT) were selectively depleted during the acute and convalescent stages of the disease. In addition, levels of immunoreactive EGF were reduced in affected submandibular glands, especially at 8 to 14 days PI with SDA virus, but some evidence of EGF depletion was seen at up to 3 weeks PI. A corresponding transient depletion of EGF receptor reactive salivary EGF was seen between 1 and 3 weeks after experimental SDA infection. These studies suggest that a clinical (or subclinical) infection with SDA virus could have significant effects on experimental studies on EGF-dependent functions, including reproductive physiology and carcinogenesis.     

Sialodacryoadenitis virus-associated lesions in the lower respiratory tract of rats

Eight- to 10-week-old outbred Wistar rats were inoculated intranasally with 10(2.9) medium mouse lethal infective doses of sialodacryoadenitis (SDA) virus. Sham inoculated control rats and challenged rats were killed at 1 day intervals for the first 8 days, then on days 10, 12, 14, and 20. Typical lesions associated with SDA were seen microscopically in the salivary and lacrimal glands of inoculated rats. In addition, laryngitis, tracheitis, bronchitis, bronchiolitis, and multifocal alveolitis were present during the acute stages of the disease. Viral antigen was demonstrated in epithelial cells lining airways by immunofluorescence microscopy. SDA virus was recovered from the lower respiratory tract from days 2 to 6 post-inoculation (PI). Serum antibodies to SDA virus, but not to Sendai virus or Mycoplasma pulmonis were present in rats tested at day 20 PI. These findings demonstrate that during the acute stages of the disease, significant lesions do occur in the lower respiratory tract of SDA virus-infected rats.     

Coronavirus infection in the laboratory rat: immunization trials using attenuated virus replicated in L-2 cells.

Sixty-nine specific pathogen-free male Wistar rats approximately eight weeks of age were used to evaluate the efficacy of an attentuated strain of sialodacryoadenitis (SDA) virus in providing protection against infection on subsequent challenge with virulent SDA virus. Fifty-four animals were inoculated intranasally with approximately 10(3.5) median cell culture infectious doses of the 25th passage of SDA virus in L-2 cells. Randomly-selected vaccinated animals were killed in order to evaluate the safety and efficacy of attenuated virus by histopathological examination of the salivary glands, lacrimal glands, and lower respiratory tract, and titration of sera for antibody to SDA virus. At three months and six months postvaccination (pv), animals were selected at random and challenged with virulent SDA virus. Seronegative, age-matched animals were also challenged, and served as controls. In animals examined at six to ten days pv, lesions were absent in submandibular and parotid salivary glands and lacrimal glands, but transient lesions were present in major airways of the lower respiratory tract. In a comparison of the incidence and extent of lesions, and antibody titers in challenged vaccinates and seronegative controls, lesions were minimal or absent in vaccinates compared to challenged naive rats, particularly in animals inoculated at three months pv. In addition, antibody titers in challenged vaccinates were much higher than were postinoculation titers in inoculated controls. In a comparison of lesions in salivary and lacrimal glands in vaccinated and control animals challenged at six months pv, there was a significant reduction in the number of animals without lesions in the vaccinated group (p = less than 0.0001).(ABSTRACT TRUNCATED AT 250 WORDS)     

Antigenic heterogeneity of sialodacryoadenitis virus isolates.

Seventeen field isolates of sialodacryoadenitis (SDA) virus had been isolated from the lung of rats with clinical SDA during epizootics of SDA from 1976 to 1978 in the research laboratory. Based on their neutralization patterns against antisera to strains 681, 930-10, Lu-3, Lu-4 and Lu-7, these isolates were divided into 3 antigenic groups. The first group consisted of 8 isolates which were neutralized by 4 out of 5 antisera at high serum dilution. The second group consisted of 6 isolates which were neutralized by only 2 antisera at high serum dilution. The third group exhibited intermediate neutralization pattern and 3 isolates belonged to it. Considering the time course of virus isolation, it was concluded that three antigenically different SDA viruses had been spread irregularly and ocassionally two had been spread simultaneously in an animal house of rats during the several epizootics of SDA.     

Coronavirus infections in the laboratory rat: degree of cross protection following immunization with a heterologous strain.

One hundred and twenty-one specific pathogen-free male Wistar rats eight to ten weeks of age were used to evaluate the efficacy of Parker's rat coronavirus (PRC) in affording cross protection on subsequent challenge with virulent sialodacryoadenitis (SDA) virus. Sixty-two animals were inoculated intranasally on day 0 and 21 days later with approximately 10(2) median tissue culture infective doses (TCID50) of the tenth passage of PRC replicated in L-2 cells. Animals were selected at random postvaccination to evaluate the safety and efficacy of PRC by histopathology, immunohistochemistry and serology. At three and six months postvaccination (PV), vaccinated and seronegative control rats were inoculated intranasally with approximately 10(3) TCID50 doses of virulent SDA virus. Challenged rats were then killed at 6, 10 and 14 days postchallenge and necropsied. Evaluations were based on lesion indices in lacrimal and salivary glands and respiratory tract, the presence of viral antigen by immunohistochemistry, and antibody response. Lesions were observed in rats killed PV, but in general, they were significantly reduced compared with those present in seronegative animals post-exposure to virulent SDA virus (p < or = 0.05). However, they were still considered to be an unacceptable level for a routine vaccination procedure. Potvaccination antibody titers to rat coronavirus were evident in all animals tested at three or six months prior to challenge with SDA virus.(ABSTRACT TRUNCATED AT 250 WORDS)     

Sequential changes in the harderian and exorbital lacrimal glands in Wistar rats infected with sialodacryoadenitis virus

A sequential light and electron microscopic study of the exorbital and Harderian lacrimal glands was done on 2.5- to 15-month-old Wistar rats exposed to sialodacryoadenitis (SDA) virus. Typical coronaviral particles were readily demonstrated in cytoplasmic vesicles of Harderian and exorbital glands examined at 6 days post-inoculation. Lesions were seen in a relatively high percentage of lacrimal glands in infected animals of all ages, with no obvious age-related variations in the incidence and extent of changes. Lesions frequently persisted for a longer interval post-exposure in lacrimal glands than in salivary glands. The persistence of lesions commonly seen in Harderian glands was attributed, at least in part, to the cytotoxic effects of porphyrin-containing secretions released during the acute necrotizing stages of the disease. The persistence of lesions in some lacrimal glands indicates that they are useful tissues for microscopic examination for the retrospective provisional diagnosis of SDA. Persistent lesions also indicate that normal functions of these glands may be compromised for up to several weeks following outbreaks of SDA.     

Rotavirus infection in foals

Fecal samples from 86 foals with diarrhea were examined by electron microscopy during a 2.5 year period. Of these, 26 (30%) were positive for rotavirus. All of the cases were found in epizootic areas. The disease was produced in an experimental foal by inoculation via stomach tube of a bacteria-free fecal filtrate containing rotavirus. Examination of postmortem tissues from the duodenum and jejunum of 2 naturally infected foals and an experimentally infected foal revealed replicating virus in the intestinal epithelial cells. A limited survey of complement-fixing antibody to rotavirus in horses from Kentucky, Virginia, and France indicated that all horses had antibody to the virus. The sole exception was 1 foal from which blood samples were collected prior to suckling. These results were presumptive evidence that rotavirus is a major cause of diarrhea in foals, and the presence of antibody in horses from diverse areas is evidence for the ubiquitousness of this infection.     

Some characteristics of a natural infection by parainfluenza-3 virus in a group of calves.

Parainfluenza-3 (Pi3) virus infection in a group of 25 calves is described. The virus was isolated from the lungs of four calve at days 6, 7, 13 and 55 after they were housed together at birth. Intracytoplasmic inclusion bodies were seen by light microscopy in bronchial and bronchiolar epithelial cells of two of these calves. Virus infected cells were detected by electron microscopy in three of the four calves. Haemagglutination inhibition antibodies to Pi3 virus were found in the sera of the calves. Despite the virus being present in the group from one week, a significant increase in antibody titre was found in only two animals although all the calves were in contact with each other during the study period. The pulmonary lesions in the four infected calves consisted of a bronchitis and bronchiolitis with infiltration of the walls and lumena of these structures by neutrophils and an adjacent neutrophil infiltration of alveoli some of which were collapsed.     

Experimental infection of young rabbits with a rabbit enteric coronavirus.

The clinical signs and lesions caused by the rabbit enteric coronavirus (RECV) were studied in young rabbits orally inoculated with a suspension containing RECV particles. The inoculated animals were observed daily for evidence of diarrhea. Fecal samples and specimens from the small intestine and from the gut associated lymphoid tissue (GALT) were collected from 2 h to 29 days postinoculation (PI) and processed for immune electron microscopy (IEM) and light microscopy. Coronavirus particles were detected in the cecal contents of most inoculated animals from 6 h to 29 days PI. Lesions were first observed 6 h PI and were characterized by a loss of the brush border of mature enterocytes located at the tips of intestinal villi and by necrosis of these cells. At 48 h PI, short intestinal villi and hypertrophic crypts were noted. In the GALT, complete necrosis of the M cells as well as necrosis of the enterocytes lining the villi above the lymphoid follicules with hypertrophy of the corresponding crypts were observed in all the animals. Five inoculated rabbits had diarrhea three days PI. The presence of RECV particles in the feces of the sick animals and the microscopic lesions observed in the small intestine suggested that the virus was responsible for the clinical signs. A few inoculated rabbits remained free of diarrhea. Fecal material collected at postmortem examination contained RECV particles. The results suggest that the virus could also produce a subclinical infection.     

Propagation of group II avian adenoviruses in turkey and chicken leukocytes

An avirulent hemorrhagic enteritis virus isolate (HEV-A) as well as a virulent one (HEV-V), both belonging to the group II avian adenoviruses, were successfully propagated in turkey leukocyte cell cultures. HEV antigens were detected as early as 12 hr after infection of the cells, using HEV-specific monoclonal antibodies in a fluorescent antibody test, and virus particles were observed by electron microscopy in the nuclei of infected cells at 18 to 24 hr after infection. Electron microscopy revealed the presence of HEV in the nuclei of nonadherent cells, as well as in adherent cells. The nonadherent infected cells had the characteristics of immature mononuclear leukocytes, whereas the adherent cells had monocyte-macrophage characteristics. HEV produced in turkey leukocytes was mostly cell-associated, particularly with the nonadherent cells. HEV-A could be serially passed in turkey blood leukocyte cultures at least seven times. Various methods employed to culture virus indicated that cells grown in spinner cultures were superior to cells grown in stationary cultures. In contrast to the successful infection of HEV in turkey leukocytes, the infection of chicken leukocytes with either HEV or splenomegaly virus of chickens, or turkey leukocytes with splenomegaly virus, was poor.     

Acute viral hepatitis in California sea lions

Acute viral hepatitis was diagnosed in five California sea lions (Zalophus californianus) stranded along the Los Angeles coast. Light microscopy revealed large nuclear inclusion bodies in hepatocytes. Electron microscopy provided evidence that these inclusion bodies were composed of adenovirus-like virions. Attempts to grow the virus in cell culture systems were unsuccessful.     

Severe leukopenia and liver necrosis in young African grey parrots (Psittacus erithacus erithacus) infected with psittacine circovirus

This paper describes the signs, clinical pathology, and postmortem findings in 14 young African grey parrots (Psittacus erithacus erithacus) that were naturally infected with psittacine beak and feather disease (PBFD) virus (psittacine circovirus). All but two of the parrots had severe leukopenia at clinical presentation. Two other parrots also had severe anemia. All birds died within 3 wk after presentation. Postmortem examination documented liver necrosis in 11 of 14 birds and secondary bacterial or fungal infections in 9 of 14 birds. Tests for Chlamydia psittaci, polyomavirus, and Salmonella sp. were negative. PBFD viral infection could be demonstrated in all birds by polymerase chain reaction. Supporting evidence of PBFD viral infection was gathered by histologic examination of the bursa of Fabricius, electron microscopy, and DNA in situ hybridization. Electron microscopic examination of both the bursa of Fabricius and liver revealed virus particles resembling circovirus. DNA in situ hybridization of six liver tissue samples confirmed the presence of PBFD virus and excluded the presence of avian polyomavirus. Our findings suggest that a specific presentation of peracute PBFD viral infection, characterized by severe leukopenia, anemia, or pancytopenia and liver necrosis in the absence of feather and beak abnormalities, may occur in young African grey parrots.     

Scanning and transmission electron microscopic study of equine infectious anemia virus.

Scanning and transmission electron microscopy were used to study in detail the morphogenesis and replication of equine infectious anemia virus (EIAV) in cultured, persistently infected equine fetal kidney fibroblasts. The EIAV was shown by thin-section electron microscopy to resemble morphologically more closely the members of the genus Lenti-virus in the family Retroviridae than other genera. Scanning electron microscopy demonstrated budding virus on only about 5% of the equine fetal kidney fibroblasts; however, the entire surface of these cells was involved in viral replication. Except where virus budding was observed, EIAV-infected cells were smooth and free of the topographic surface alterations characteristic of cells transformed by type C retroviruses. The morphologic relationship of EIAV and pathologic manifestations of EIAV infection to those of other Retroviridae are discussed.     

Lymphoid tissue tropism of porcine reproductive and respiratory syndrome virus replication during persistent infection of pigs originally exposed to virus in utero

The ability of porcine reproductive and respiratory syndrome virus (PRRSV) to establish a persistent infection is the principal contributing factor to the world-wide spread of the disease. Several studies have documented the course of viral infection in postnatally infected pigs; however, very little is known regarding sites of virus replication during persistent infection of pigs exposed to PRRSV in utero. In this study, virus replication and PRRSV-specific antibody were followed for several hundred days in a group of pigs derived from three sows infected at 90 days of gestation with PRRSV isolate VR-2332. Eighty-four percent of pigs were born viremic with a mortality of 54% within 21 days after birth. At approximately 60 days sera from pigs were negative for virus by virus isolation. Analysis of virus replication in the tissues of pigs randomly sacrificed between 63 and 132 days showed no evidence of virus in lung and other non-lymphoid organs. However, virus was easily recovered from tonsil and lymph nodes and in situ hybridization identified these tissues as sites of virus replication. Even though replication was at a low level, virus was easily transmitted to sentinel pigs. By 260 days pigs became seronegative and did not transmit virus to sentinel pigs. Sacrifice of remaining pigs after 300 days showed no evidence of virus in blood and tissues. This study shows that congenital PRRSV-infected pigs can support virus replication for an extended period during which virus replication is primarily restricted to tonsil and lymph nodes.     

Importance of the medullary macrophage in the replication of lymphoid leukosis virus in the bursa of Fabricius of chickens

Electron microscopy and immunocytochemistry were used to study the development of lymphoid leukosis virus infection in the bursa of Fabricius of experimentally infected chicken embryos and chickens. In embryos infected at 7 days of incubation and killed 10 days later, virus particles and group-specific viral antigen were confined mainly to the connective tissue of the lamina propria of the bursal mucosal folds; a few developing follicles had discrete virions and group-specific antigen between cells. In chickens infected at 1 day of age, infection (as determined by use of electron microscopy and immunocytochemistry) was maximal in 1- to 4-month-old birds, and the greatest concentration of virus and group-specific viral antigen was in the medulla of the follicles. Although lymphoid leukosis virus was released from lymphocytes, epithelial cells, and macrophages, virus replication in the medullary macrophages was more active than that in the other cells. Normal medullary macrophages had cell membrane vesicles (50 to 80 nm in diameter) that covered part of all of the cell membrane surface. In infected chickens, virus particles frequently developed within these vesicles. Comparable vesicles were not found on cortical macrophages. Results of the present study indicated that the medullary macrophage was the principal host cell for replication of lymphoid leukosis virus in the bursa of Fabricius of the chicken.     

1. Isolation and Characterisation of Equine Rhinopneumontis Virus and Other Equine Herpesviruses from Horses

Equine herpesviruses (EH viruses) were isolated from 9 horses in three separate outbreaks of respiratory disease. The pattern of disease in the three stables is described and evidence is presented that some of the horses were ill, possibly as a result of recurrent infection, and that reactivation of a persistent, latent infection may have occurred. An ulcerative condition of the pharyngeal region was seen in some of the horses with EH virus infection.
The cytopathogenicity for equine foetal kidney cells of the 9 EH viruses varied considerably. One isolate, EH 39 virus, which was recovered from an acute, upper respiratory tract infection, was rapidly cytopathic for equine foetal kidney cell cultures and was shown in neutralisation tests to be identical with, or closely related to equine rhinopneumonitis virus (EH virus type 1) that is associated with acute respiratory disease and abortion in other countries. More slowly cytopathic isolates were recovered from mild to subclinical upper respiratory tract infections. Evidence is presented that the property of slow cytopathogenicity is probably related to the tendency of these viruses to remain cell associated. Slowly cytopathic isolates were recovered from the nasal cavity of horse 89 on two occasions 79 days apart. One of the eight slowly cytopathic isolates, EH 86 virus, was shown to be antigenically distinct from equine rhinopneumonitis virus (EH 39 virus).     

Seroepizootiology of types 3 and 7 adenoviruses and bovine viral diarrhea virus infections of beef cattle from birth to first parturition

Twenty-nine female Angus cattle were monitored from birth until 24 months of age for infection with types 3 and 7 adenoviruses (AV3, AV7) and bovine viral diarrhea (BVD) virus by virus isolation and neutralization tests. Twenty-seven animals remained in the study for 450 days, whereas 16 animals remained at the conclusion of the tests. Viruses were not isolated from test animals. Seroepizootiologic examination indicated that there were 28 infections with AV3, 33 infections with AV7, and 13 infections with BVD virus. Of these infections, 5 with AV3, 6 with AV7, and 2 with BVD virus were reinfections. All but 1 primary AV3 infection had occurred between 180 and 360 days after parturition. Infection with AV7 occurred earlier than that with AV3, as shown by 6 seropositive animals by 180 days. Most adenovirus infections were documented after animals had left confinement and were turned out to pasture. Most BVD virus infections occurred within the first 90 days after parturition. At completion of the study, only 1 animal had not shown evidence of AV3 infection, 2 animals had not been infected with AV7, and 7 cows had remained free of BVD virus infection.     

Serological studies of parainfluenza type 3 virus, bovine adenovirus type 3 and bovine respiratory syncytial virus infection in beef calves

Six serum samples were taken at monthly intervals from birth to weaning from each of 41 newborn calves in the autumn and spring calf crops of a beef cow--calf herd. The serum hemagglutination-inhibition (HI) antibody titres to parainfluenza type 3 virus (PIV-3), virus-neutralization (VN) antibody titres to bovine adenovirus type 3 (BAV-3) and bovine respiratory syncytial virus (BRSV) were determined using microtitration techniques. There was serological evidence of a significantly higher incidence of infection with BAV-3 in the fall calves than in the spring calves. Serological responses to BAV-3 were not detected in calves with VN titres of greater than 1/256. Serological evidence of subclinical infection with PIV-3 occurred mainly in late February or early March during a period of marked environmental temperature fluctuations. Serological evidence of a high incidence of infection with BRSV was obtained for both the fall and spring calf crops. Serum antibody appeared to be unable to prevent infection with BRSV. An association between infection with BRSV and clinical pneumonia was found in 3 out of 9 calves. BAV-3 infection was related to pneumonia in only 1 instance; however, there was simultaneous evidence of BRSV infection in this calf. PIV-3 infection was found to be related to pneumonia in only 1 instance. There was serological evidence of infection with BAV-3 in association with the occurrence of diarrhea in 3 calves.     

Persistence of FDMV and its effects on disease control strategies

It is well-known that foot-and-mouth disease virus (FMDV) causes a persistent infection, lasting for more than 28 days, in cattle, sheep, goat as well as some other ruminant species, but not in pigs. Although convincing evidence for virus transmission is missing, these carrier animals have to be considered as a potential risk of infection. Some aspects of FMDV persistence are presented and discussed with regard to disease control strategies.