The effect on newborn chicks of oral inoculation of reovirus isolated from chickens with tenosynovitis

Reovirus strain 176, isolated from chickens with tenosynovitis, was highly pathogenic following oral inoculation of 1-day-old specific-pathogen-free chicks. Disseminated lesions including hepatic necrosis, splenic lymphostromal cell hyperplasia, and bursal atrophy occurred on day 3 postinoculation (PI), followed by myocarditis on day 6 PI and by pericarditis and tenosynovitis on day 9 PI. Reovirus was isolated from the liver as early as day 1 PI, whereas significant neutralizing antibody was detected on day 13 PI. Mortality occurred from day 4 to day 7 PI, and the death of birds was associated with the severity of hepatic necrosis. The occurrence of tenosynovitis in virus-inoculated birds was subclinical.     

An atypical strain of Pasteurella gallinarum: pathogenic, phenotypic, and genotypic characteristics.

The pathogenicity of a strain of Pasteurella gallinarum isolated in Fresno County, Calif., was compared with the American Type Culture Collection (ATCC) strain. Broiler chickens were inoculated intranasally with 10(7) colony-forming units (CFU) and intramuscularly with 10(5) CFU of each strain. The only notable lesions were in chickens inoculated intramuscularly with 10(5) CFU of the Fresno strain, which developed severe myositis at the inoculation site, pericarditis, perihepatitis, airsacculitis, and synovitis. P. gallinarum was reisolated from these lesions. Phenotypic characteristics of the two strains were identical except in reactions in ONPG broth and fermentation of xylose. Protein-banding patterns for the two strains were identical except for a single band difference in the 35-kilodalton region. Restriction endonuclease analysis confirmed that the Fresno strain was a distinct one. Plasmid analysis revealed that the ATCC strain had two plasmids and the Fresno strain had none.     

Prevalence of reoviruses in commercial chickens

The prevalence of reoviruses in commercial chickens with the runting/stunting syndrome, tenosynovitis, and normal chickens was investigated. Reoviruses were isolated from 3-week-old chickens affected with the runting/stunting syndrome and from older chickens with tenosynovitis; viruses were isolated from tissues with and without lesions. Reoviruses were also frequently isolated from rectal contents of normal 3-week-old chickens, and there was serological evidence of previous reovirus infection in all flocks of adult meat breeder chickens examined. The widespread occurrence of reoviruses in both normal and diseased chickens indicates that the isolation of reoviruses from tissues of chickens with lesions does not necessarily imply any aetiological relationship of reovirus with disease, even in the absence of other known causes. However, the occurrence of reovirus in normal chickens does not preclude an aetiological relationship with disease and further investigation of strain variation and possible virulence factors in avian reoviruses is required.     

Experimental gizzard erosions in specific-pathogen-free chicks by serotype 1 group I avian adenoviruses from broilers

The pathogenicity of serotype 1 group I avian adenovirus (GIAAV) strains isolated from gizzard erosions for 1-wk-old specific-pathogen-free (SPF) chickens was investigated. In Experiment 1, SPF chickens inoculated orally with a serotype 1 strain (Tokushima2000/GE) of GIAAV isolated from gizzard erosion in broilers were euthanatized and necropsied from 3 to 12 days after inoculation. Clinically, there were no significant signs in the chickens inoculated with Tokushima strain. The gizzards were found macroscopically to be dilated with liquefied contents. The koilin layer of the gizzard exhibited multifocal white lesions with occasional erosions. Gross lesions of the koilin layer (white foci and erosion) appeared at 3 days after inoculation and persisted until 12 days after inoculation. The gross lesions were most severe at 9 days after inoculation. Histologically, multifocal degeneration, necrosis, and loss of the glandular epithelial cells with intranuclear inclusions, liquefactive change of koilin substance, and erosive lesions of koilin layers and surface epithelium of the gizzards were observed. Macrophages increased in the focal areas of the lamina propriabelow the liquefactive koilin substances. Immunohistochemically, intranudear inclusions were positive against GIAAV antigen. Histologic lesions of gizzards were most severe at 7 and 9 days after inoculation, but the frequency of intranuclear inclusions was greatest at 5 days after inoculation. In Experiment 2, the pathogenicities of four strains of serotype 1 GIAAV for 1-wk-old SPF chickens were compared. Three strains isolated from gizzard erosion had similar pathogenicities but one strain from a normal chicken was not pathogenic for chickens. These results suggest that gizzard erosions by serotype 1 GIAAV infection may occur in broiler chickens in Japan.     

Virulence and toxigenicity of capsular serogroup D Pasteurella multocida strains isolated from avian hosts

Five capsular serogroup D strains of Pasteurella multocida isolated from avian hosts were examined for virulence and toxigenicity. Virulence was based on development of lethal infections or lesions following intramuscular exposure of turkey poults. The four strains isolated from turkeys varied from slightly to moderately virulent; the strain isolated from a chicken was avirulent. Poults exposed by intra-airsac inoculation with relatively few organisms of the more virulent of the strains had a high mortality rate; however, intranasal exposure of poults with this strain did not cause clinical disease or establish infections. All strains from turkeys were toxigenic, producing heat-labile toxins that killed poults when administered intraperitoneally and caused focal dermal lesions when administered intradermally. Using these criteria, the strain from a chicken was not toxigenic. The demonstration of virulence, particularly the high mortality in poults exposed via air sacs, indicates avian capsular serogroup D strains are a potential cause of fowl cholera.     

Cross protection of mice and swine given live-organism vaccine against challenge exposure with strains of Erysipelothrix rhusiopathiae representing ten serovars

Mice and swine vaccinated (subcutaneous inoculation) with live acriflavine-fast attenuated Erysipelothrix rhusiopathiae, strain Koganei 65-0.15 (serovar 2), were challenge exposed with 10 strains of E rhusiopathiae pathogenic for swine; the latter strains comprised serovars 9 and 10 and other previously undetermined. Vaccinated mice did not die after they were challenge exposed (subcutaneous inoculation) with serovars 4, 6, 7, 8, 9, 10, 15, 16, or N, but vaccinated mice challenge exposed with strain 2553 (serovar 20) had 30% mortality. Nonvaccinated control mice died after they were challenge exposed with all serovars tested. One of 2 vaccinated swine challenge exposed (intradermal inoculation) with each of strains 911 (serovar 8), 2179 (serovar 10), or 2553 developed localized urticarial lesion at the site of intradermal inoculation. Vaccinated swine challenge exposed with serovars 4, 6, 7, 9, 15, 16, or N did not have clinical signs of acute swine erysipelas. Nonvaccinated control swine developed localized lesions at the site of intradermal challenge inoculation.     

Enteropathogenicity of Dutch and German avian reoviruses in SPF white leghorn chickens and broilers

The enteropathogenicity of avian reoviruses (ARVs), isolated from chickens affected with malabsorption syndrome (MAS) from The Netherlands and Germany was studied. In the first trial seven different ARVs isolated from MAS cases were inoculated in 1-day-old specific pathogenic free (SPF) white leghorns. The pathogenicity was compared with 2 ARVs isolated from cases of tenosynovitis, namely reference strain S1133 and a Dutch strain. Although a difference in the severity of the clinical disease was observed, all reoviruses could induce vacuolar degeneration and sloughing of the epithelium of the small intestine at day 2 post inoculation (PI) till day 7 PI. Two Dutch and one German ARV derived from MAS causing the most severe intestinal lesions at day 2 PI, were further studied in the second trial using SPF broilers. These reoviruses did not cause weight gain depression in the broilers although lesions in the small intestine were present from day 1 up to day 4 PI and were more severe than in the white leghorn chickens. In one of the inoculated groups apical denuded villi were already present at day 1 PI. At day 7 PI the small intestine of the infected broilers appeared to be normal. Reovirus antigen was detected in the cytoplasm of the enterocytes at the tip and middle section of the affected villi both in layers and in broilers. To study the role of intestinal CD4+ and CD8+ T-cells and macrophages/monocytes in the pathogenesis of ARV, the numbers of these cells of the jejunal villi of one infected and the control broiler groups were compared. CD4+ T-cells were detected in low numbers and only in the infected broiler group at day 14 PI. The numbers of CD8+ T-cells and macrophages/monocytes were significantly higher in the infected broiler group than in the control broiler group at day 7 and 14 PI and at day 7 PI respectively. Our study indicates that the reovirus alone cannot induce intestinal lesions as found in MAS chickens. Moreover, CD8+ T-cells may play a major role in the pathogenesis and or reovirus clearance in the small intestine.     

Virulence and transmissibility of Mycoplasma gallisepticum

The virulence of 4 low passage strains of Mycoplasma gallisepticum obtained from different sources within Australia was studied by experimental infection of chickens. Strain Ap3AS, originally isolated from the air sac of a broiler chicken, produced severe air sac lesions following injection into the abdominal air sacs of 2-week or 3-week-old chickens, and adult hens. Strain 80083 which was isolated from a clinically normal broiler breeder hen was also capable of producing gross air sac lesions following intra-abdominal (IA) injection, although it did so less consistently than strain Ap3AS. Strain 82078 isolated from a layer hen and strain QXO which was isolated from a turkey were also moderately pathogenic in terms of the incidence and severity of lesions elicited following IA injection. Strains Ap3AS and 80083 both caused a substantial loss of egg production over a 5 week period after IA infection of 27-week-old hens. Neither strain Ap3AS nor 80083 caused gross lesions or loss of egg production when administered alone into the upper respiratory tract. However, when inoculated into the conjunctival sac in combination with the Vic S strain of infectious bronchitis virus (IBV) strains Ap3AS and 80083 produced identical clinical signs of conjunctivitis. The mean numbers of M. gallisepticum in tracheal washings were significantly higher 2 weeks after infection in the group receiving strain 80083 in combination with IBV than in the group infected with strain Ap3AS and IBV (p less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)     

Characterization of avian reovirus strain-specific polymorphisms

Avian reoviruses have been associated with several pathologic conditions, but correlative relationships between genotypes and specific diseases have not been demonstrated. Six avian reoviruses (883, 176, 81-5, S1133, FC, and TX) were selected for this study, and a comparative study of the pathogenic properties of the viruses in chickens, following peroral and footpad inoculation, was carried out, along with a comparison of the electrophoretic mobility of viral genomic segments and viral proteins encoded by the gene segments. The pathogenic properties of the viruses were shown to be diverse, with three distinct pathotypes being defined: Pathotype I (883) caused only a syndrome that we have termed "transient digestive system disorder" (TDSD); Pathotype II (FC, TX, and S1133) caused only "viral arthritis syndrome" (VAS), whereas Pathotype III (176 and 81-5) caused both TDSD and VAS. Likewise, the genomes of the viruses were shown to be extremely polymorphic, with a maximum of five segments co-migrating between any two strains. Considerable variation in the electrophoretic mobility of the encoded proteins also was demonstrated with pronounced variation in the molecular size of the sigma 4 protein, the purported viral attachment protein, being evident. These results show that the genomes of avian reoviruses were extremely polymorphic, preventing correlation between genotypes and pathotypes. But these studies have provided us with the genetic elements needed to characterize the gene functions involved in viral pathogenesis.     

Avian colibacillosis caused by an intestinal pathogenic Escherichia coli isolate from calf diarrhea

An intestinal pathogenic Escherichia coli isolate from calf diarrhea, containing the iutA, f17A, afa-8D, and cnf2 genes, was able to cause avian colibacillosis after experimental infection in chickens. Intra-tracheal inoculation and spray of this strain caused 10% of mortality and gross lesions, including airsacculitis, pericarditis, and perihepatitis. These results suggest that some bovine pathogenic E. coli can cause extra-intestinal infections in other animal species.     

Pathogenicity of serotype 8 fowl adenovirus isolated from gizzard erosions of slaughtered broiler chickens

The pathogenicity of serotype 8 fowl adenovirus (FAV), isolated from gizzard erosions of slaughtered broiler chickens, was investigated. In experiment 1, 29 5-day-old specific-pathogen-free (SPF) chickens were inoculated with the isolates of serotype 8 FAV, M013 (group 1) or G0054 (group 2) strain, via an oral route. There were no clinical signs in any of chickens after inoculation, and mild gizzard erosions were observed macroscopically and microscopically in three inoculated chickens of group 2. FAV was recovered from gizzards and rectums but was not recovered from pancreas and livers from chickens in both inoculated groups. In experiment 2, 27 1-day-old SPF chickens were inoculated with the G0054 strain by intramuscular route. Five, 6, and 3 inoculated chickens died on days 3, 4, and 5 postinoculation (PI), respectively. Four, 3, 1, and 1 inoculated chickens became moribund with severe clinical signs such as ruffled feathers, severe depression and closed eyes from days 3 to 6 PI, respectively. Macroscopically, the common characteristic of the gross lesions of dead chickens and euthanized moribund chickens was discoloration of liver. FAV was recovered from the gizzard, liver, pancreas and rectum. Virus titers in the liver and pancreas were high until day 6 PI. Histologically, necrotizing hepatitis and pancreatitis with intranuclear inclusion bodies were observed in the inoculated chickens. These results indicate that some strains of serotype 8 FAV are able to reproduce not only gizzard erosion by oral inoculation but inclusion body hepatitis (IBH) by intramuscular inoculation.     

PATHOLOGY AND TUBERCULIN SENSITIVITY IN CATTLE INOCULATED WITH MYCOBACTERIUM AVIUM COMPLEX SEROTYPES 6, 14 AND 18

SUMMARY Three strains of Mycobacterium avium complex organisms, serotypes 6, 14 and 18 isolated from typical tuberculous lesions in cattle were examined for pathogenicity and ability to sensitise cattle to avian and bovine tuberculin. Each strain caused tuberculoid granulomas at the site of subcutaneous inoculation but no lesions elsewhere. Sensitisation to bovine tuberculin was detected in the caudal fold test in 11 of 18 inoculated animals 8 weeks after injection. In a simultaneous comparative cervical test, reactions to avian tuberculin were much larger than reactions to bovine tuberculin in all inoculated animals.     

Pathologic study of specific-pathogen-free chicks and hens inoculated with adenovirus isolated from hydropericardium syndrome.

The mortality and pathology caused by serotype 4 adenovirus, isolated from chickens with hydropericardium syndrome (HPS) in Japan, was investigated in specific-pathogen-free (SPF) chickens. One-day-old to 15-mo-old SPF chickens were inoculated intramuscularly, orally, and intranasally with liver homogenates from HPS chickens or isolated serotype 4 adenovirus. There were no clinical signs before death. The mortality rate in all groups of 1-day-old chicks was 100%, irrespective of the inoculum or inoculation route. Four-week-old chickens inoculated with liver homogenate also had a 100% mortality rate. Five-week-old chickens inoculated with cell culture of HPS adenovirus had a 40% mortality rate. The mortality rates of 7-mo-old hens inoculated with liver homogenates intramuscularly and orally were 75% and 25%, respectively. In 15-mo-old hens inoculated with liver homogenates intramuscularly, the mortality rate was 70%. Gross lesions were hydropericardium and swelling and congestion of the liver with occasional petechial hemorrhages. Histologically, the liver had diffuse or multifocal hepatic necrosis and hemorrhage with intranuclear inclusion bodies noted within hepatocytes. In the spleen, macrophages containing erythrocytes and yellow pigment were prominent in the red pulp. In the lung, a moderate diffuse macrophage infiltration was noted throughout the lung parenchyma, and these macrophages contained yellow pigment. In the pancreas of the chicks inoculated at 1 day old, there was multifocal necrosis of glands with intranuclear inclusion bodies. Intranuclear inclusion bodies were seen also in the gizzard, proventriculus, duodenum, cecum, kidney, and lung of the chicks inoculated at 1 day old. Immunohistochemically, the intranuclear inclusion bodies of various organs showed positive reactions against group I avian adenovirus. Adenovirus was recovered from the liver of chickens with HPS. This study indicates that HPS adenovirus is able to reproduce HPS lesions and mortality in SPF chicks and even adult chickens and that it is a highly pathogenic strain.     

Kidney lesions associated with mortality in chickens inoculated with waterfowl influenza viruses

Seventy-six type A influenza viruses recovered from waterfowl in Wisconsin, California, South Dakota, Florida, Texas, Alabama, and Nebraska were tested for virulence in chickens. The challenge to chickens was intravenous inoculation of first-, second-, or third-egg-passage virus. Each of the virus strains was tested separately in three or four chickens. Eighteen of the 76 viruses caused the death of one or more chickens following inoculation. Postmortem lesions were similar in all dead birds. In decreasing order of frequency, gross lesions included: swollen kidneys evident as accentuated lobular patterns, urates in the pericardial sac, and urates on the surface of the liver. Microscopic lesions present in kidneys were consistent with visceral gout. Mortality was associated with inoculations having higher concentrations of infectious virus. These results indicate that the influenza A viruses circulating in duck populations may include strains potentially pathogenic for chickens.     

鸡病毒性关节炎的病原分离与鉴定

从疑似鸡病毒性关节炎的病鸡趾屈肌腱鞘中,用7日龄SPF鸡胚经卵黄囊接种,分离出一株病毒。该病毒株经电镜观察,病毒粒子呈六角形、二十面体对称的双层衣壳结构,核心直径约75nm,无囊膜。该病毒不受DNA抑制剂的影响,核酸型为RNA病毒;在pH 4.0～8.0保持稳定;能抵抗56℃ 20h;抵抗乙醚作用1h;1％的福尔马林30h可使其灭活;病毒感染力为:ELD_(50)为10~(-2.87)/0.1 mL,TCID_(50)为10~(-3.65)/0.1mL。用该病毒经呼吸道接种1日龄雏鸡,可引起跖关节肌腱水肿,生长发育缓慢,接种后20d死亡率达70％。血清学检查,发病鸡群30d时琼扩阳性率为72％;人工接种雏鸡21d时琼扩阳性率为100％。根据实验结果,结合流行病学及临床症状,确定病原为鸡病毒性关节炎病毒(VAV)。     

Adenoviral gizzard erosion in commercial broiler chickens

Pathologic and immunohistochemical changes caused by group I of the fowl adenovirus (FAV) serotype-1 99ZH strain, isolated from broiler chickens exhibiting gizzard erosion, were investigated in commercial broiler chickens. One hundred twenty-two chickens were inoculated with the strain by both oral and ocular routes at 1, 3, or 5 weeks of age and euthanatized for necropsy within 4-18 days of inoculation. Focal gizzard erosions were observed in the inoculated chickens of each age group. A histologically degenerative koilin layer, necrotic mucosa, intranuclear inclusion bodies in the glandular epithelial cells, inflammatory cell infiltrations in the lamina propria, submucosa, and a muscle layer were seen in the gizzards. Immunohistochemical staining showed evidence of FAV antigens in the intranuclear inclusion bodies. These findings were recognized regardless of their maternal antibody levels for FAV serotype-1. Gizzard lesions appeared later in the lower-dose-inoculated chickens than in the higher-dose-inoculated chickens. Numerous CD3-positive cells and IgY-positive plasma cells were seen in the gizzard lesions. In 5-week-old chickens the heterophil infiltrations in the lesions were milder than in younger chickens. Intranuclear inclusion bodies also were observed in the epithelial cells of the ileum or cecal tonsils of some chickens. Thus, this study shows that FAV-99ZH causes adenoviral gizzard erosion in broiler chickens without hepatic or pancreatic lesions and that cell infiltration is more severe than in dietary gizzard erosions.     

Comparative microscopic lesions in reoviral and staphylococcal tenosynovitis

Experimental inoculation of 1-day-old male broiler chickens with avian reo-virus or Staphylococcus aureus caused tenosynovitis of the gastrocnemius and digital flexor tendons. Reovirus inoculation by either the oral or footpad route initiated a diffuse lymphocytic infiltration in the peritendineum, synovial membrane, and epitenon from 1 to 5 weeks postinoculation (PI). Heterophils were not a predominant feature of the inflammatory response, but when present they were localized with fibrin in and around synovial spaces. The prevalence of microscopic tendon lesions was less common with staphylococcal infection than with reovirus infection. With staphylococcus, lesions were localized to the synovial space and membranes and were characterized by heterophils and fibrin but few lymphocytes. Synovial cell hyperplasia and bursal atrophy were common in both groups. From 10 to 20 weeks PI, both groups developed progressive tendon fibrosis. These results indicate that tenosynovitis due to inoculation with reovirus or staphylococcus may be differentiated histologically from 1 through 5 weeks PI. After 10 weeks, this may not be possible, because diffuse fibrosis was the major lesion with both. Perhaps fibrosis predisposes older, heavier broilers to tendon failure and rupture.     

Role of route of exposure, age, sex, and type of chicken on the pathogenicity of avian reovirus strain 81-176

Chickens were evaluated by age, sex, and type for susceptibility to reovirus strain 81-176 inoculated subcutaneously. Chicks were most susceptible to the lethal effects of reovirus infection at hatching, after which resistance increased rapidly. By 1 week of age, mortality was negligible, but chicks were still susceptible to the less lethal effects of the virus. Mortality rates of males and females were equal. Leghorn and broiler-type chicks did not differ appreciably in their response to viral inoculation. An effort was made to find a more "natural" means of exposure to reovirus than parenteral inoculation. Neither oral nor aerosol exposure was as effective as subcutaneous inoculation. Attempts to transmit reovirus to susceptible hatching chicks, starting when they were in ovo (19 days of incubation), also failed. Decreased weight gain proved to be a valid criterion for judging reovirus infection. Reovirus infection lowered the mean values of body weights, and the standard deviations were substantially greater, indicating an unevenness in size of the affected chickens.     

Molecular characterization of a virulent genotype VIId strain of Newcastle disease virus from farmed chickens in Shanghai

A virulent Newcastle disease virus strain was isolated from diseased chickens in Shanghai, China. The isolated strain was initially characterized as highly virulent because of a short mean death time in embryonated chicken eggs and specific-pathogen-free chickens and was typed as neurotropic by intracloacal inoculation of chickens. The isolated strain had a dibasic amino acid motif in the fusion protein cleavage site sequence required for systemic replication in the host cell. The strain fell into subgenotype VIId by phylogenetic analysis of the fusion protein gene. Although these results demonstrated some sequence similarity between the isolated strain and strains responsible for outbreaks of Newcastle disease in China and Taiwan, the unusually high mortality (86.4%) set this strain aside from other VII strains. Finally, a cross-protection assay demonstrated that La Sota and clone 30 live vaccines could not protect chickens from infection with the isolated strain, with a zero survival rate being observed when chickens were challenged with a high dose of virulent VIId virus.     

Experimental reovirus hepatitis in newborn chicks.

The avian reovirus "UM 1-203" originally isolated in the United States from chickens with tenosynovitis was pathogenic for the newborn chick infected by parenteral inoculation. It induced plurivisceral lesions, which became particularly intense in the liver. The intense local multiplication of the virus provoked a necrotizing hepatitis; viral titers were maintained around an E.I.D.50 of 10(8)/0.2 ml of organ suspension in chicks killed between the 3rd and 5th days after inoculation. As a specific cellular response to the viral multiplication, numerous polykaryocytes formed and increased in number, then regressed and disappeared from the hepatic parenchyma. By histologic and electron microscopic examination at progressive times after infection, the virus was recognizable in the polykaryocyte cytoplasm. In chicks that survived longer (killed the 5th and 6th days after inoculation), regeneration of the hepatic parenchyma occurred and seemed to develop from groups of dense hepatocytes that either lacked the virus or survived the acute phase of infection.