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1.
Striped skunks (Mephitis mephitis) were inoculated into the right submandibular salivary gland with street rabies virus. They were killed at various times after inoculation and several tissues were examined by immunofluorescence and light microscopy. Right and left superior cervical, nodose and trigeminal ganglia, medulla oblongata and at least three regions of right and left submandibular salivary glands were examined by the fluorescent antibody technique. Intracerebral titrations of salivary gland suspensions were made in weanling white Swiss mice. Immunofluorescent material (inoculum) was detected in septa and connective tissue surrounding secretory units of the right submandibular gland immediately after inoculation, but otherwise antigen was not detected in either right or left submandibular glands without coincident antigen in the medulla oblongata. This occurred first on day 12 in areas of the gland remote from the inoculation site. Titers of virus were low at this time. Serum neutralizing antibodies occurred by day 7 in a few skunks. The time of development and distribution of antigen strongly suggest that, even after direct inoculation, neural networks are necessary for development of widespread infection of the salivary gland. The early occurrence of serum neutralizing antibodies in some of the skunks suggests that the immune response was activated by virus in the inoculum since immunofluorescence was not detected in any tissue at this time.  相似文献   

2.
Yolk sacs (YS) were examined by gross, histochemical, and electron microscopic techniques from incubation day 12 to 10 days of age. Weights were maximal at 22 days of incubation and declined markedly thereafter; by 7 days of age, the yolk was completely absorbed. Endodermal cells of YS epithelium were highly polar with nuclei, mitochondria, and glycogen in basal areas and lipid globules and intralysosomal yolk granules near luminal surfaces. Yolk laminar spherules were first seen at day 16 of incubation. They increased progressively with incubation and disappeared rapidly after hatching. Spherules stained for calcium, glycosaminoglycans, and alkaline phosphatase in late incubation and were considered calcium storage structures. Histochemical evidence of intracellular calcium loading of yolk matrix (from 18 days of incubation until 5 days of age) suggested a specific calcium transport function of YS epithelium.  相似文献   

3.
Following the infection of turkey poults with a field isolate of infectious bursal disease virus, antibody levels were examined and reisolation of the virus was attempted. After inoculation at 36 days of age, peak titres in both the inoculated and a contact-exposed group were obtained after 13 days. The titres fell slightly during the next week and then remained level until the experiment was terminated at 91 days of age. Virus was reisolated from faeces from day 3 until day 8 after inoculation in the inoculated group and from day 4 until day 9 in the contact-exposed group. In the inoculated group, virus was recovered from the bursae, spleens and intestines on both days 4 and 6 after inoculation, but the thymuses on day 6 only. No clinical signs were observed.  相似文献   

4.
Diaphragms obtained from 40 fetal rabbits at gestational ages of 20, 22, 25 and 30 days were examined by light and transmission electron microscopes. The percentage of myogenic cells undergoing mitosis was calculated by counting mitotic nuclei in 1 micron sections. At 20 and 22 days gestation, myogenic cells (myoblasts and satellite cells) were actively proliferating with 2.66 +/- 0.41% (n = 15) and 2.18 +/- 0.20% (n = 23), of the cells in division, respectively. The myogenic cells undergoing mitosis appeared to be of both myoblasts and satellite cells at these stages of the development. The myotubes on day 20 of gestation contained a large number of lipid droplets and an abundance of glycogen particles which were reduced by day 22 of gestation. The mitotic rate on day 25 of gestation was reduced to 1.16 +/- 0.11% (n = 28). The muscle fibers were well differentiated at this stage and the majority of the dividing cells were considered to be satellite cells. In diaphragms from the full term fetuses (day 30), the mitotic rate was reduced to 0.31 +/- 0.05% (n = 24). It was suggested that myoblasts of the fetal rabbit diaphragm proliferated for the myotube formation during the earliest stage of the development (day 20 of gestation) and then the number of satellite cells increased after day 22 of gestation. Growth of the organ after day 25 of gestation appeared to be mostly due primarily to the hypertrophy and differentiation of the muscle fibers rather than proliferation of myoblasts.  相似文献   

5.
The present study describes the pathogenesis of infection of chicks with a new avian reovirus strain, belonging to the so-called enteric reovirus strains (ERS) that is capable of causing central nervous system signs in SPF white leghorns. After intramuscular (IM) or oral inoculation birds were either observed for clinical signs or sacrificed for macroscopic, histological and virological examination for 21 days. Virus isolation was performed on the brain, leg muscle, hock joint, liver and spleen. For the detection of viral antigen the immunohistochemistry (IHC) technique was performed on the caudal part of the cerebrum, spinal cord including spinal ganglia and right N. Ischiadicus. High mortality (79% in 7 days) was seen in birds that were inoculated IM. Survivors were depressed and stayed small until the end of the experiment. One bird had tremor and showed torticollis at 9 days after IM inoculation. Birds that were inoculated orally were depressed from day 4 and stayed small until the end of the experiment. One bird showed a torticollis at 10 days after inoculation. After both IM and oral inoculation ERS was isolated from the brain between 3 and 10 days after inoculation. Other examined organs were positive for virus isolation from day 1 or 5 until day 21. IHC revealed viral antigen positive cells in the Plexus chorioideus (plexus epithelial cells or cells within the underlying connective tissue) and in a spinal ganglion. The results indicate that the pathogenesis of ERS infection in chickens bears some resemblance with that of the mammalian reoviruses serotype 1 in mice.  相似文献   

6.
A fatal encephalomyelitis was developed after intracerebral and hind limb inoculation of in 6-week-old C57BL/6J mice by the inoculation of fixed rabies virus (CVS-11 strain), intracerebrally and into hind. After the intracerebral inoculation, virus antigens were detected in the cerebral cortex and hippocampus at 2 days postinoculation (PI), and later spread centrifugally to thalamus, brain stem, cerebellum, spinal cord and spinal ganglia. At 4 days PI, severe apoptosis and DNA fragmentation were observed in the hippocampus and cerebral cortex. All mice infected intracerebrally were dead without limb paralysis at from 10 to 11 days PI. In contrast, mice infected with virus intramuscularly were persistently observed virus antigens in the myocytes at the site of inoculation from 2 days PI. At 4 days PI, the antigens were demonstrated in the spinal dorsal root ganglia, spinal cord and muscle spindles without their detection in the cerebrum and hippocampus. There were no apoptosis in the spinal cord and dorsal root ganglia, however hind limb paralysis was found in all infected mice. Hind limb paralysis was progressed to quadriparalysis, and mice were dead from 11 to 13 days PI. From 4 days PI, necrosis of neuron was observed in the the spinal and dorsal ganglia with infiltration of lymphocyte. This study suggested that the necrosis of spinal neurons was more important to cause the paralysis of hind limb rather than the severe cerebral infection and apoptosis in C57BL/6J mice infected with CVS-11 strain. The virus primarily replicated in the muscles was ascended the spinal cord via afferent fibers and retrogradely invaded the cerebrum, and with subsequent spread to muscle spindles.  相似文献   

7.
The development and transmission of Anaplasma marginale was studied in Dermacentor andersoni males. Laboratory-reared male D andersoni were allowed to feed for 7 days on a calf with ascending A marginale parasitemia. The ticks were then held in a humidity chamber for 7 days before being placed on 2 susceptible calves. Anaplasmosis developed in the calves after incubation periods of 24 and 26 days. Gut and salivary glands were collected from ticks on each day of the 23-day experiment and examined with light and electron microscopy. Colonies of A marginale were first observed in midgut epithelial cells on the sixth day of feeding on infected calves, with the highest density of colonies found in gut cells while ticks were between feeding periods. The first colonies contained 1 large dense organism that subsequently gave rise to many reticulated organisms. Initially, these smaller organisms were electron-lucent and then became electron-dense. On the fifth day after ticks were transferred to susceptible calves for feeding, A marginale colonies were found in muscle cells on the hemocoel side of the gut basement membrane. A final site for development of A marginale was the salivary glands. Colonies were first seen in acinar cells on the first day that ticks fed on susceptible calves, with the highest percentage of infected host cells observed on days 7 to 9 of that feeding. Organisms within these colonies were initially electron-lucent, but became electron-dense.  相似文献   

8.
OBJECTIVE: To evaluate the ultrastructural changes and localization of encephalomyocarditis virus (EMCV) and viral pathogenesis in the myocardium of experimentally infected piglets. ANIMALS: Eight 20-day-old piglets. PROCEDURE: Six piglets were inoculated oronasally with 5 ml (10(6) median tissue culture infective dose/ml) of EMCV suspension, and 2 were used as uninfected controls. Piglets were euthanatized or died between postinoculation days 1 and 3. Samples of heart tissue from all piglets were evaluated histologically, by virus isolation, and by use of immunohistochemistry and electron microscopy. RESULTS: All infected piglets had gross or microscopic lesions of interstitial myocarditis. immunohistochemically, EMCV antigen was detected in the cytoplasm of cardiac muscle cells, Purkinje fibers, and endothelial cells and in the nucleus of cardiac muscle cells and Purkinje fibers. Ultrastructural lesions were characterized by degeneration and necrosis of cardiac muscle cells and Purkinje fibers. Virus was present intracytoplasmically in cardiac muscle cells, Purkinje fibers, and endothelial cells of capillaries and intranuclearly in cardiac muscle cells. The cell membranes of the Purkinje fibers and endothelial cells had distinct protrusions that contained virus particles. In control piglets, no lesions were found, and no EMCV antigen was detected. CONCLUSIONS: Localization of EMCV intracytoplasmically or intranuclearly in various myocardial cells may well reflect the sites of viral proliferation. The presence of virus particles in cell membrane protrusions and in vacuoles within the lumen of capillaries indicates that virus is released not only by disintegration of the host cell but also via exocytosis.  相似文献   

9.
Inclusion bodies, indistinguishable from rabies inclusion bodies (Negri bodies), were found in the brains of 8 nonrabid dogs. The inclusions were compared to Negri bodies present in neurons of rabies-positive animals and examined for the presence of rabies virus by a combination of immunoperoxidase staining (7 cases), fluorescent antibody (FA) staining (1 case), and transmission electron microscopy (4 cases). Positive immunoperoxidase staining for rabies was obtained in brain tissues from FA rabies-positive animals. All brain tissues from the 7 dogs stained by the immunoperoxidase method and the brain from the 1 dog stained by the FA method were negative for rabies. Rabies virus was not found in inclusion-containing neurons in the cases examined by transmission electron microscopy. These results emphasize the importance of FA testing and mouse inoculation for the diagnosis of rabies.  相似文献   

10.
Studies of ERA/BHK-21 rabies vaccine in skunks and mice.   总被引:5,自引:5,他引:0       下载免费PDF全文
ERA rabies vaccine virus grown in BHK-21 13S cells (ERA/BHK-21) and street rabies virus were titrated in mice by intracerebral, intranasal and intramuscular inoculation. Mice were also given undiluted ERA/BHK-21 in baits. Skunks were given undiluted ERA/BHK-21 in baits and by intramuscular, intranasal and intestinal inoculation. Virus neutralizing antibody titers against rabies virus were measured over a three month observation period. The surviving skunks were challenged by intramuscular inoculation with rabies street virus from a skunk salivary gland suspension. When titrated in mice, ERA/BHK-21 had titers of 10(7.0), 10(5.2) and 10(3.9) median lethal doses per mL by the intracerebral, intranasal and intramuscular routes, respectively. All skunks (8/8) inoculated intranasally developed paralytic rabies by 12 days after exposure to ERA/BHK-21 virus. None of the skunks that developed vaccine-induced rabies had infectious virus in the submandibular salivary glands. Vaccine-induced rabies also occurred in 1/8 skunks in the intramuscularly inoculated group and in 1/8 in the intestinally inoculated group. The survival rates of challenged skunks in the various groups were as follows: intramuscular, 7/7; intestinal, 2/7; bait, 0/8; and control, 0/8. These results indicate that ERA/BHK-21 virus has a significant residual pathogenicity in mice and in skunks by some routes of inoculation. Skunks given vaccine intramuscularly were protected against challenge, while those skunks given the vaccine in baits were not.  相似文献   

11.
Infectious bovine rhinotracheitis virus was rapidly cleared from the nasal mucosa of calves after intranasal aerosol exposure. Nonimmune calves (experiment 1) cleared 10(9) plaque-forming units (PFU) of virus from the nasal mucosa in less than 4 hours and 10(6) PFU of virus in 1 hour. An eclipse phase followed the clearance of viral inoculum. Replicating virus was first detected at 9 hours. Viral titers increased stepwise until maximum was attained on postinoculation day 4. Virus persisted in the nasal mucus until day 12. Clinical signs of disease corresponded with the shedding of virus. In contrast to nonimmune calves, immune calves (experiment 2; same calves as in experiment 1, but 30 days after initial exposure) cleared 10(9) PFU of virus in 1 hour and 10(6) PFU of virus in less than 5 minutes. An abortive reinfection occurred after exposure of immune calves with 10(9) PFU of virus. Virus was first detected in these calves at 14 hours after exposure and was not detected beyond 24 hours after inoculation. Immune calves given 10(6) PFU of virus did not shed virus after clearance of inoculum. Clinical signs of infection were not observed in immune calves after viral challenge exposure. The date indicated that there was no detectable residual virus beyond 3 hours after the exposure.  相似文献   

12.
Six calves inoculated intranasally with a vaccinal strain of infectious bovine rhinotracheitis (IBR) virus and 6 control calves were given a placebo. All calves were subsequently challenge exposed (by aerosol) with rhinovirus--3 of the calves from each group at 2 days after they were inoculated with IBR virus or with placebo and the remaining calves at 6 days. Nasal excretion of viruses, interferon (IFN) concentrations in nasal secretions (NS), and neutralizing antibody in sera and NS were determined. All calves given the vaccinal IBR virus subsequently had IFN in their NS. Interferon was detected as early as 1 day, reached maximal titers at 2 to 4 days, and persisted in individual calves for 5 to 10 days after inoculation. Rhinovirus shedding was not detected from IBR virus-inoculated calves whose NS contained both rhinovirus antibody and IFN at the time of challenge exposure; such calves were protected at either 2 or 6 days after IBR virus inoculation. The outcome of rhinovirus challenge exposure of calves whose NS contained IFN, but not rhinovirus antibody, varied with the day of challenge exposure. Rhinovirus excretion was detected from 2 of these calves challenge exposed 2 days after IBR virus inoculation, but was not detected from a calf challenge exposed 6 days after inoculation. However, while IFN was present in NS from the former 2 calves, rhinovirus shedding was markedly reduced as compared with that from control calves without IFN or NS antibody at the time of challenge exposure. Consistent relationship was not observed between the rhinovirus neutralizing antibody titer of calves' sera and NS. The antibody titer of NS more closely correlated with protective immunity to rhinovirus infection than did the serum antibody titer.  相似文献   

13.
Ferrets experimentally infected by various routes with pseudorabies virus were examined for gross and microscopic lesions. Nonsuppurative meningoencephalomyelitis, as well as visceral lesions, occurred. The incubation period seemed related to the viral dose and to the distance between the inoculation site and the central nervous system. The distribution of the lesions in the central nervous system appeared to be closely related to the peripheral nerve pathways from the inoculation sites. Other findings indicated that the lymphohematogenous route could have a role in the dissemination of the virus in infected ferrets.  相似文献   

14.
应用建立的Nested PCR特异地检出狂犬病病毒株CVC、HEP-Flury、ERA、RC-HL、1008、Komatsug-awa的RNA,但对类狂犬病病毒Lagos bat、Duvenhage、Mokola及水泡性口膜炎病毒、轮状病毒、犬瘟热病毒均为阴性。该法敏感性很高,能检出3 TCID50或0.8pg的狂犬病病毒RNA。用该法测定了小鼠脑内感染CVS株后的病毒增殖和移行动态,对感染小鼠的主要内脏器官进行了病毒RNA检测,结果发现小鼠脑内感染CVS 5 d以后在其心、肝、脾、肺等内脏器官均检出了病毒RNA。  相似文献   

15.
Postexposure prophylaxis for prevention of rabies in dogs   总被引:1,自引:0,他引:1  
OBJECTIVE: To evaluate postexposure prophylaxis (PEP) in dogs experimentally infected with rabies. ANIMALS: 29 Beagles. PROCEDURE: Dogs were sedated and inoculated in the right masseter muscle with a salivary gland homogenate from a naturally infected rabid dog (day 0). Six hours later, 5 dogs were treated by administration of 2 murine anti-rabies glycoprotein monoclonal antibodies (mAb) and commercial vaccine; 5 received mAb alone; 5 received purified, heat-treated, equine rabies immune globulin (PHT-ERIG) and vaccine; 5 received PHT-ERIG alone; 4 received vaccine alone; and 5 control dogs were not treated. The mAb or PHT-ERIG was administered at the site of rabies virus inoculation. Additional vaccine doses for groups mAb plus vaccine, PHT-ERIG plus vaccine, and vaccine alone were administered IM in the right hind limb on days 3, 7, 14, and 35. RESULTS: All control dogs and dogs that received only vaccine developed rabies. In the PHT-ERIG and vaccine group, 2 of 5 dogs were protected, whereas none were protected with PHT-ERIG alone. Use of mAb alone resulted in protection in 4 of 5 dogs. Administration of mAb in combination with vaccine provided protection in all 5 dogs. CONCLUSIONS AND CLINICAL RELEVANCE: Current national guidelines recommend euthanasia or a 6-month quarantine for unvaccinated animals exposed to rabies. Findings from this study document that vaccine alone following severe exposure was unable to provide protection from rabies. However, vaccine combined with mAb resulted in protection in all treated dogs, revealing the potential use of mAb in PEP against rabies in naive dogs.  相似文献   

16.
Pseudorabies virus was inoculated into the uterus of 15 gilts within 6 hours after natural breeding, and gilts were necropsied at postbreeding days (PBD) 3, 6, 10, 14, and 28; 3 control gilts were treated similarly, except for inoculation with pseudorabies virus and were necropsied at PBD 6, 10, and 14. Tissues were collected for virus isolation, fluorescent antibody staining, and histopathologic examination. Pseudorabies virus was isolated from the reproductive tract up to day 14. Lesions in the reproductive tract consisted of multifocal to diffuse lymphohistiocytic vaginitis and endometritis, and lymphoplasmacytic aggregates in the corpora lutea. Multiple ulcers were seen in the vagina or endometrium of several gilts at PBD 3, 6, and 10. Corpora lutea of 1 gilt were necrotic at PBD 14 and contained large numbers of inflammatory cells. Focal aggregates of lymphocytes and plasma cells were seen in vagina and endometrium of 3 gilts and in the ovary of 1 gilt at day 28.  相似文献   

17.
A tissue culture infection test in routine rabies diagnosis.   总被引:3,自引:0,他引:3       下载免费PDF全文
A cell culture infection test was developed for the isolation of rabies virus from field cases submitted for rabies diagnosis. The procedure involved the addition of a suspension of suspect brain tissue to a suspension of murine neuroblastoma cells in 96-well microtiter plates. The cultures were then incubated at 35-36 degrees C for four days at which time they were fixed, stained with a fluorescein-labelled hamster antirabies antibody conjugate and examined with a fluorescence microscope. Rabies antigen in cells was readily visible as brilliant, apple-green fluorescent particles. This technique was compared with the standard mouse inoculation test and was at least as sensitive to infection with small amounts of virus, required a much shorter test period and was substantially more economical than the mouse inoculation test. The new cell culture test is now in use at this laboratory, replacing the mouse inoculation test.  相似文献   

18.
Chick embryos infected with Akabane virus by the yolk sac route at 6 days of incubation developed polymyositis and encephalitis. At 3 to 7 days after inoculation, skeletal muscles had myotubule degeneration, clumping of muscle cell nuclei, and infiltration of heterophils; dysplasia and aplasia were evident at 9 to 15 days after inoculation. Changes in the cerebral neostriatum and optic lobes at 2 to 11 days after inoculation included necrosis of primordial nervous tissue, hemorrhages, and hyperplasia of the vascular endothelial cells. Cavities were in nervous tissue subsequent to encephalitis. Hydranencephaly and vascular wall thickening were found 13 and 15 days after inoculation. Embryos infected intravenously at 15 days incubation had foci of encephalitis 3 to 6 days after inoculation, including neuronal degeneration, neuroglial hyperplasia, vascular endothelial proliferation, and heterophil infiltration.  相似文献   

19.
Fatal herpesvirus infection in commercial rabbits   总被引:1,自引:0,他引:1       下载免费PDF全文
Acute mortality occurred in two unrelated rabbitries. In the rabbits examined, an unidentified herpesvirus caused lesions that have not been reported previously in this species. The primary lesions were multifocal hemorrhagic dermatitis on the face and back, localized pneumonia, and severe splenic necrosis. Large eosinophilic, intranuclear inclusion bodies that were observed in tissue sections of skin, spleen, and lung were identified as herpes-like viral particles by electron microscopy, and herpesvirus was cultured on rabbit kidney cells. Intramuscular injection of tissue culture fluid containing virus resulted in mortality and severe illness in two seven-week-old domestic rabbits four and six days postinfection, respectively. The gross and microscopic lesions were reproduced and herpeslike viral inclusions were observed in skin lesions. Herpesvirus was recovered from lung, trachea, spleen, liver, and from the thigh muscle at the site of inoculation. The experimental infection also activated severe pasteurella septicemia. The herpesvirus isolate needs further characterization.  相似文献   

20.
Groups of two or three day old pigs were inoculated intravenously with cell culture grown transmissible gastroenteritis virus. A single or a multiple dosage schedule was used. The magnitude of immune response was measured in terms of serum neutralization indices. A single dose of relatively attenuated virus caused mild clinical signs of transmissible gastroenteritis infection in the pigs and induced a low level of antibody in the serum by the seventh day after inoculation. Repeated injections of virus at seven day intervals stimulated little increase in antibody titers. However, high serum antibody titers were obtained for all pigs if the time interval between injections was extended to 15 days. Sera obtained early after exposure to live transmissible gastroenteritis virus contained mainly IgM antibody whereas sera obtained later after exposure contained mainly IgG antibody. Ten plaque purified isolates of transmissible gastroenteritis virus, comprising eight American isolates, one Japanese isolate and one British isolate were indistinguishable by means of reciprocal plaque reduction neutralization tests.  相似文献   

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