Pathologically and serologically different avian nephritis virus isolates implicated in etiology of baby chick nephropathy.

Three virus isolates (WG-3, -4, and -5) from chicks affected by baby chick nephropathy were orally inoculated into 1-day-old specific-pathogen-free chicks of lines PDL-1 and 15I. Additional chicks were orally inoculated with avian nephritis virus (ANV) strain G-4260. Chicks inoculated with isolates WG-3, -4, and -5 died between 2 and 6 days postinoculation (PI), with mortality ranging from 0% to 53.3%. Pathological findings in the dead chicks included nephrosis in chicks inoculated with WG-3, -4, and -5, and nephritis and visceral urate deposition in chicks inoculated with G-4260. The stability of the WG-5 isolate, as well as the size of the particles and the nucleic acid type, were also similar to those of the G-4260 strain. All of the examined chicks inoculated with WG-3, -4, and -5 had interstitial nephritis at 14 days PI. Therefore, the three virus isolates were considered to be ANV. However, there was no serological relationship between the isolates and ANV (G-4260 and M-8 strains).     

Experimental infection in specific-pathogen-free chicks with avian reovirus and avian nephritis virus isolated from broiler chicks showing runting syndrome

Avian reovirus (ARV) and avian nephritis virus (ANV) were individually isolated from runty 10-day-old broiler chicks. The ARV isolate, IR-R, the ANV isolate, IR-N, and the reference strain of ANV, G-4260, were inoculated orally into 1-day-old chicks of two specific-pathogen-free (SPF) chicken lines, 151 and PDL-1. Growth retardation without the presence of gross lesions was clearly observed at 7 and 14 days postinoculation (PI) in chicks of both lines inoculated with the IR-R strain. On the other hand, in chicks inoculated with IR-N strain, growth retardation was observed only in the chicks of line 151 at 7 and 14 days PI. Microscopically, nephritis was observed in both chicken lines at 7 and 14 days PI. When chicks that were inoculated with the IR-N strain at 1 day of age were inoculated with the IR-R strain at 3 days of age, growth retardation was observed in the chicks of line PDL-1 at 10 and 17 days PI. However, the growth retardation was less severe than in the group receiving a single inoculation of the IR-R strain.     

Differences in the induction of urate deposition of specific-pathogen-free chicks inoculated with avian nephritis virus passaged by five different methods.

The G-4260 strain of avian nephritis virus (ANV) was passaged using five different methods as follows: method 1, passage three times in chorioallantoic membrane (CAM) of 11-day-old embryonated eggs (CAM3); method 2, passage twice in CAM and further passage once in yolk sac (YS) of 6-day-old embryonated eggs (CAM2-YS1); method 3, passage 11 times in CAM (CAM11); method 4, passage 10 times in CAM and further passage twice in YS (CAM10-YS2); method 5, passage as in Method 4 and then passage three times in chicken kidney cell culture (CAM10-YS2-CK3). CAM11 and CAM10-YS2 were each inoculated orally into 25 one-day-old specific-pathogen-free (SPF) chicks. Seven chicks in the CAM11-inoculated group and six chicks in the CAM10-YS2-inoculated group died or were killed because they were moribund; all had either nephrosis or urate deposition. CAM3, CAM2-YS1, CAM10-YS2, and CAM10-YS2-CK3 were each inoculated intraperitoneally into 15 one-day-old SPF chicks. No chicks inoculated with CAM3 or CAM2-YS1 died, but wo chicks inoculated with CAM10-YS2 and three inoculated with CAM10-YS2-CK3 died with urate deposition. At 14 postinoculation, plasma urate values of the CAM10-YS2- and CAM10-YS2-CK3-inoculated chicks were significantly higher than those of CAM3- and CAM2-YS1-inoculated chicks and control chicks (P less than 0.01). However, interstitial nephritis was observed in most of the ANV-inoculated birds.     

Experimental factors affecting mortality following inoculation of chickens with avian nephritis virus (G-4260).

Groups of approximately 20 one-day-old chickens were inoculated with G-4260, the reference strain of avian nephritis virus (ANV), or saline. Based on mortality rates from severe nephritis in comparable experiments, light Sussex chickens generally were more susceptible than Rhode Island red (RIR) chickens. Mortality was greater in those given broiler starter than those given other feeds, and was greater when light Sussex chickens were given broiler starter feed and cold-stressed at 15 +/- 1 C for 2 hr daily during the first week rather than brooded normally. Inoculation with G-4260 either orally or by intraperitoneal injection produced similar results in RIR chickens. Thirty-three inoculated chickens died of severe nephritis between 4 and 12 days postinoculation, and 24 (73%) of them had visceral urate deposits. Inoculated inbred white leghorn Line 15 chickens with maternal antibody to ANV were brooded normally and given broiler feed: they were susceptible to infection as evidenced by subsequent histological lesions in the kidneys and serology, but mortality was not a feature. There were no deaths from nephritis in inoculated non-inbred white leghorn chickens free of maternal antibody to ANV that were given broiler feed and brooded normally. These results have implications in standardizing experimental conditions for the study of mortality induced by G-4260 and similar viruses.     

Visceral urate deposits in chicks inoculated with avian nephritis virus

Avian nephritis virus, G-4260 strain, was inoculated orally into one-day-old specific-pathogen-free chicks of two lines. Approximately 20 per cent of the chicks of both lines died with visceral urate deposits from eight to 12 days after infection, and the virus was isolated from the kidneys of the dead chicks. At 14 or 15 days of age the mean liveweight of the surviving infected chicks was approximately 16 per cent less than that of the uninfected control chicks.     

The pathogenesis of Pasteurella multocida serotype A:3,4 infection in turkeys: a comparison of two vaccine strains and a field isolate

The pathogenesis of avian pasteurellosis caused by two vaccine strains, M-9 and Clemson University (CU), and a highly virulent field isolate, 86-1913, of Pasteurella multocida (serotype A:3,4) was studied in 7-week-old turkeys inoculated by an oculo-nasal-oral technique. Turkeys inoculated with strain CU and isolate 86-1913 developed severe progressive bacteremia that began at 4 hours postinoculation (PI) and peaked at 16-20 hours PI. Turkeys inoculated with strain CU and isolate 86-1913 had significantly higher concentrations of bacteria in blood and tissues, and greater histologic lesion scores for necrosis, heterophil infiltrates, and intralesional bacteria than turkeys inoculated with strain M-9. Immunohistochemical staining specific for P. multocida demonstrated numerous extracellular bacteria in tissues from turkeys inoculated with strain CU and isolate 86-1913. The mortality for turkeys inoculated with isolate 86-1913 was significantly higher than for turkeys receiving the two vaccine strains.     

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.     

Induction of hydropericardium in one-day-old specific-pathogen-free chicks by adenoviruses from inclusion body hepatitis

The pathogenicities of inclusion body hepatitis (IBH) and hydropericardium syndrome (HPS) strains of adenovirus for specific-pathogen-free (SPF) chicks were compared. One-day-old SPF chicks inoculated intramuscularly with the DPI-2 (serotype 2), S-PL1 (serotype 2), TR630 (serotype 8), and Saga97 (serotype 8) strains from IBH and with the LVP-1 strain (serotype 4) from HPS exhibited the mortality, liver enlargement, and hydropericardium characteristic of gross change found in HPS. The chicks inoculated with the IBH and HPS strains exhibited similar histologic and immunohistochemical changes. Neither mortality nor pathologic changes occurred in 3-wk-old SPF chicks inoculated with IBH strains, although HPS strain induced HPS lesions in them. This study indicates that IBH strains of adenovirus can also reproduce HPS lesions and mortality in 1-day-old SPF chicks and that IBH and HPS strains may have similar pathogenicities except for their different virulence for older chickens.     

Evaluation of pathogenicity and protective efficacy of serotype 2 Marek's disease virus from birds belonging to genus Gallus in Japan

The new cloned serotype 2 Marek's disease viruses (MDV) of ML-6, ML-9, and ML-22 strains were inoculated in specific-pathogen-free (SPF) chicks to evaluate the pathogenicity and protective efficacy. Chicks inoculated or contact-infected with ML strains showed no gross and histological lesions in lymphoid organs, sciatic plexuses and other visceral organs during 10 weeks of observation periods, indicating that the viruses were non-pathogenic. Moreover, the viruses were found to be spread horizontally among chicks by demonstrating the presence of viremia in contacted chicks at 2 weeks-old. Chicks vaccinated with ML-6 at one day-old were protected against subsequent challenge by inoculation with virulent MDV strain of Md/5 at 4 or 7 days old or by contact infection at 7 days old with chickens previously inoculated with the same strain.     

Effects of cyclophosphamide in newly hatched chickens after inoculation with avian nephritis virus

Effects of immunosuppression were compared in newly hatched chickens given cyclophosphamide (CY) after inoculation with avian nephritis virus (ANV). All CY-treated infected chickens died within 13 days after inoculation of the virus and had heavy urate deposits throughout the body. However, non-CY-treated infected, CY-treated noninfected, and non-CY-treated noninfected control chickens survived through the observation period. In a chronologic study, the value of serum uric acid in CY-treated infected chickens was more than 3 times higher than that in non-CY-treated infected chickens, and more than 9 times higher than in noninfected chickens. Serum uric acid values were coincident with the positive degree of ANV antigen in the tubular epithelial cells in the kidneys and with the severity of renal degeneration. Serologic and immunohistologic examinations did not reveal detectable antibody and IgG- and IgM-containing cells in the spleen and kidneys of CY-treated infected chickens. However, non-CY-treated infected chickens had an increased number of IgM- and IgG-containing cells and antibody against ANV on postinoculation day 6. These findings demonstrated that CY treatment enhanced the susceptibility of chickens to ANV infection.     

Serologic and pathogenetic studies on avian reoviruses isolated in Japan

Eighty-nine avian reoviruses isolated from diseased and clinically normal chickens were classified serologically using antisera against five prototype strains. Eighty-three strains were classified into five serotypes; six strains were untypable. Most of the cytopathogenic strains that produced a clear cytopathic effect (CPE) in chicken embryo fibroblasts (CEFs) were highly pathogenic for chicken embryos (80% or more mortality via the allantoic sac) and for chicks (severe footpad swellings and tenosynovitis). These strains were classified into a single serotype represented by the TS-142 prototype strain. However, 10 strains that could not produce a clear CPE in CEFs showed very low pathogenicity for embryos and chicks, and these strains were serologically different from the TS-142 prototype strain. There was a strong relationship between pathogenicity and serotype. About 17% of the isolates were considered highly pathogenic.     

Serological differentiation of avian infectious bronchitis field isolates using an enzyme immunoassay: presence of Dutch strains in West Germany

Six field virus isolates were identified as the etiologic agent of avian infectious bronchitis (IB) in West Germany. A serological comparison was carried out using the M-41 strain from the Massachusetts serotype and the Dutch variant strains D-207, D-1466, D-3128, and D-3896 in a cross indirect immunoperoxidase test. Three isolates demonstrated similarity to the M-41 strain; the remainder showed a closer antigenic relationship to the Dutch variant strains. These results correspond to differences observed in a hemagglutination-inhibition test. The enzyme immunoassay is proposed as another possible method for differentiating IB virus strains.     

传染性法氏囊病病毒变异株的致病性

传染性法氏囊病病毒变异ＧＣ９０２株可引起１０日龄ＳＰＦ鸡胚肝脏坏死和脾脏明显肿大及较高的死亡率。接种２周龄ＳＰＦ雏鸡,同时设标准Ｉ型强毒ＣＪ８０１株和标准变异株强毒１０８４Ａ株接种对照,该毒株接种后第３天才出现法氏囊粘膜浆膜出血、个别黄化、质硬等病变,且于第４天法氏囊明显萎缩变小,至接种后２０天法氏囊仍严重萎缩;接种后第２天引起脾脏显著肿大,于第１２天时大小恢复正常。试验结果表明,该ＧＣ９０２株对ＳＰＦ雏鸡的致病性与标准变异株基本一致,仅有一定的差异,但明显不同于标准Ｉ型强毒株的致病性。     

Comparative virulence of NAD-dependent and NAD-independent Actinobacillus pleuropneumoniae strains.

The virulence of a NAD-independent Actinobacillus pleuropneumoniae serotype 2 strain and NAD-dependent serotype 2, 3 and 9 strains was compared under experimental conditions. Hysterectomy-derived piglets were inoculated endobronchially with 50-500 cfu of these strains. All 23 piglets inoculated with the NAD-dependent strains developed acute disease within 12 hours post inoculation. Twenty-two of these piglets died within 24 hours after the first clinical signs. Three of nine piglets inoculated with the NAD-independent strain did not develop clinical disease. In the other six piglets, disease signs were similar as in the piglets inoculated with the NAD-dependent strains. No differences in clinical disease were observed between colostrum deprived piglets and piglets that obtained colostrum from a SPF sow.     

Detection of avian nephritis virus in Australian chicken flocks

Avian nephritis virus (ANV) is thought to infect poultry flocks worldwide, but no confirmed case has been reported in Australia. The first such case is described in this study. Cases of young chickens with clinical signs of dehydration and diarrhea were submitted to our laboratory and histopathology detected interstitial nephritis. Vaccine strains of infectious bronchitis virus were detected in some of these cases but were not considered to be the causative agent. A total of seven fresh submissions from broiler chicken flocks were collected at 8-11 days of age. Degenerate PCR primers were designed based on published ANV polymerase gene sequences and used to analyze historic cases as well as the fresh submissions. Six of the seven fresh submissions, and one historic case, were positive for ANV with nucleotide sequencing confirming these results. These results establish ANV as an infectious pathogen circulating in Australian poultry.     

The early pathogenesis in chicken inoculated with non-pathogenic serotype 2 Marek's disease virus.

A newly cloned serotype 2 Marek's disease virus (MDV), strain ML-6, was inoculated via the nasal cavity in specific-pathogen-free chicks to examine early virus replication and the expression of Marek's disease (MD)-related antigens. Following inoculation, viral intracellular antigens (VIAs) were detected in lymphoid organs (bursas and spleens) between 5 and 14 days post inoculation (PI), in feather follicles between 14 and 30 days PI, and in lungs at 3 days PI by the immunohistopathological staining of avidin-biotin-peroxidase complex method. But, very few VIAs were expressed in the thymuses between 5 and 14 days PI. However, MD tumor-associated surface antigens were not detected in any organs. Viruses were isolated from separated spleen cells at 14 and 30 days PI. Fluorescent antibodies of convalescent sera were also detected after 10 days PI. As most of the VIAs were detectable in B-cells in bursas and spleens. B-cells were considered to be the main first target cells for the serotype 2 MDV infection.     

产毒素多杀性巴氏杆菌的鉴定

采用菌落多重PCR方法对分离保存的28株多杀性巴氏杆菌进行种型和毒素基因的检测,结果表明,菌株C51-6、M-4和P-2237为产毒素多杀性巴氏杆菌,菌株C51-6和P-2237为荚膜血清D型,菌株M-4为荚膜血清A型。同时用金黄色葡萄球菌抑制试验、中性吖啶黄沉淀试验和豚鼠皮肤坏死试验对PCR方法进行了验证。基于对甘露醇、卫茅醇、山梨醇、海藻糖的发酵能力和产生鸟氨酸脱羧酶的特性,3株菌株鉴定为多杀性巴氏杆菌多杀亚种。     

Serological studies of Actinobacillus (Haemophilus) pleuropneumoniae strains of serotype 6 and their antigenic relationship with other serotypes

Serological tests such as agglutination, coagglutination, precipitation and indirect haemagglutination were used to study the antigenic relationship of reference and field strains of Actinobacillus (Haemophilus) pleuropneumoniae of serotype 6 with reference strains of other serotypes. Both cell-associated particulate and cell-free soluble antigens prepared from unheated and heat-treated bacterial suspensions of reference and field strains of serotype 6 were used in the studies. Species-specific, common antigenic determinants associated mainly with heat-treated particulate antigens of serotype 6 were cross-reactive in tube agglutination tests with almost all the serotypes. The species-specific antigens were of a minor nature because the cross-reactivities were abolished in both 2-mercaptoethanol agglutination and coagglutination tests. Cell-free saline extracts of both unheated and heat-treated suspensions of serotype 6 strains possessed epitopes specific for serotypes 3, 5 and 8 in addition to their own specific determinants. The epitopes were dominant because the reactions of strains of serotype 6 with antisera against serotypes 3, 5 and 8 persisted in almost all the serological tests used. Serotype 6 strains were antigenically closer to serotype 8 than to serotypes 3 or 5. A combination of serological tests such as coagglutination followed by 2-mercaptoethanol tube agglutination and, or, immunodiffusion tests differentiated serotype 6 strains from those of other cross-reacting serotypes.     

Serotypes of Yersinia pseudotuberculosis recovered from domestic livestock

The serological identity of 234 strains of Yersinia pseudotuberculosis recovered from domestic animals and birds in New Zealand was determined by slide agglutination test. Thirty strains were also examined by tube agglutination test. The strains were isolated from cattle (56), sheep (8), deer (117), goats (13), pigs (7), rabbits (6), guinea pigs (5), and aviary species of birds (22). All strains were isolated from animals or birds which had died or shown signs of ill health and amongst which diarrhoea was a common feature. Serotype I accounted for 23% (53) of strains, serotype II for 13% (30) of strains and serotype III for 64% (151) of strains. It was concluded that further investigations on the prevalence and serological identity of strains recovered from clinically healthy animals mav provide useful information in assessing the significance of various serotypes as a cause of disease in livestock.