One-day-old SPF chicks were inoculated with the Cux-l strain of chicken infectious anemia virus (CIAV), and the clinical development of disease and its macroscopic and microscopic alterations in the thymus and bone marrow, were observed. Tissue sections of thymus and bone marrow were stained using the streptavidin-biotin peroxidase method and examined under light microscope for evaluation of antigenic intensities in tissues. Those findings were then compared with blood parameters and ELISA results obtained through collected sera during sacrifice procedures. We sought to determine: the localization of viral antigens in thymus and bone marrow tissues after inoculation, the correlation between antigen intensities and hematologic, serologic and histopathologic findings, definitive diagnostic criteria using histopathologic and immunoperoxidase methods, and the reliability of these methods in the diagnosis of CIAV infection. For this purpose, 83, one-day-old SPF chicks were used. The birds were divided into experimental (n = 52) and control (n = 26) groups. A virus dose of TCID50 of 100,000/ml was administered intramuscularly to every bird in the experimental group. Based on the results of this study, we have suggested that clinical examination, along with macroscopic and microscopic evaluation of the thymus and bone marrow, maybe undertaken starting from day 7 post-inoculation (PI). ELISA, might be of value, as it might give consistent results starting from day 14 PI. However, the most reliable results were obtained through examination of thymus and bone marrow sections from infected birds stained by immunoperoxidase technique, as early as day 4 PI.
Animals ; Bone Marrow/*pathology ; *Chicken anemia virus ; Chickens ; Circoviridae Infections/pathology/*veterinary ; Immunoenzyme Techniques ; Poultry Diseases/*pathology ; Specific Pathogen-Free Organisms ; Thymus Gland/*pathology

In order to observe the effect of the immune and weight of chickens after use the attenuated vaccine with low dose of chicken infectious anemia virus (CIAV). In this study, the effects of low dose of CIAV on the weight of SPF chickens and NDV antibody production were observed by simulated experiments. The results showed that 10 EID50 and 5 EID50 CIAV per plume attenuated NDV vaccines were used to cause the weight loss of SPF chickens. Compared with the use of the non contaminated vaccine group, it has significant difference. And NDV antibody levels compared with the use of the non contaminated groups also decreased after use the vaccine with two doses of CIAV contaminated. It has significant difference. A certain proportion of CIAV antibody positive was detected at the beginning of the second week after use the NDV vaccine with two doses of CIAV contaminated. The detection of a high proportion of CIAV nucleic acid was detected in the first week after the use of a contaminated vaccine. The results of the study demonstrate the effects of CIAV pollution on the production and immune function of SPF chickens, and it is suggested that increasing the detection of viral nucleic acid can help save time and improve the detection rate in the detection of exogenous virus contamination by SPF chicken test method.
Animals ; Antibodies, Viral ; immunology ; Chicken anemia virus ; genetics ; immunology ; physiology ; Chickens ; Circoviridae Infections ; immunology ; veterinary ; virology ; Poultry Diseases ; immunology ; virology ; Specific Pathogen-Free Organisms ; Vaccines, Attenuated ; administration & dosage ; genetics ; immunology

Eight 2-day-old SPF chickens were each inoculated orally with a single dose of 5+O105 oocysts of Cryptosporidium baileyi, and immunoglobulin G (IgG) antibody responses were chronologically measured by indirect immunofluorescent antibody (IFA) assay. Anti-C. baileyi IgG antibody levels remained high(1:106.67 to 1:512.00) for at least 4 months with 330 days of a detectable period. Ten days after the negative conversion, each chicken was re-challenged with 1+O107 oocysts of the same species. Subsequent infection in 340-day-old individuals caused sudden elevated IgG antibody levels and the titer peaked on day 28 postchallenge inoculation(PCI), at 1:1,024 with a 65 days of detection period. Chickens in primary infection showed oocyst shedding profiles, but did not exhibit any oocyst shedding before or after experimental reinfection.
parasitology-protozoa ; Cryptosporidium baileyi ; chicken ; IgG ; immunology

Chicken anemia virus (CAV) is an important viral pathogen that causes anemia and severe immunodeficiency syndrome in chickens worldwide. In this study, a potential diagnostic monoclonal antibody against the CAV VP1 protein was developed which can precisely recognize the CAV antigen for diagnostic and virus recovery purposes. The VP1 gene of CAV encoding the N-terminus-deleted VP1 protein, VP1Nd129, was cloned into an Escherichia (E.) coli expression vector. After isopropyl-beta-D-thiogalactopyronoside induction, VP1Nd129 protein was shown to be successfully expressed in the E. coli. By performing an enzyme-linked immunoabsorbent assay using two coating antigens, purified VP1Nd129 and CAV-infected liver tissue lysate, E3 monoclonal antibody (mAb) was found to have higher reactivity against VP1 protein than the other positive clones according to the result of limiting dilution method from 64 clones. Using immunohistochemistry, the presence of the VP1-specific mAb, E3, was confirmed using CAV-infected liver and thymus tissues as positive-infected samples. Additionally, CAV particle purification was also performed using an immunoaffinity column containing E3 mAb. The monoclonal E3 mAb developed in this study will not only be very useful for detecting CAV infection and performing histopathology studies of infected chickens, but may also be used to purify CAV particles in the future.
Animals ; Antibodies, Monoclonal/biosynthesis/genetics/*immunology ; Antigens, Viral/analysis ; Capsid Proteins/genetics/*immunology ; Chicken anemia virus/genetics/*immunology ; *Chickens ; Circoviridae Infections/blood/immunology/*veterinary/virology ; Escherichia coli/genetics ; Immunohistochemistry/veterinary ; Liver/virology ; Mice ; Mice, Inbred BALB C ; Microscopy, Fluorescence/veterinary ; Poultry Diseases/blood/immunology/*virology ; Specific Pathogen-Free Organisms ; Thymus Gland/virology

Using PCR technique, the vp3 gene of chicken anemia virus (CAV) was cloned into the eukaryotic expression vector pcDNA3 to construct a recombinant pcDNA-vp3. Restriction enzyme digestion and sequencing analysis revealed that CAV vp3 gene was correctly inserted into the blank vector pcDNA3. After LipofectAMINE-mediated transfection in vitro with pcDNA-vp3 and pcDNA3 respectively, the total mRNA was extracted from liver carcinoma cell lines HepG2 and diploid cell line L-02, and RT-PCR was performed afterward. The results of RT-PCR suggested that vp3 gene was expressed in these two cell lines. At the same time, using in situ apoptotic detection assay, TUNEL kits, the apoptotic cells were found in pcDNA-vp3 transfected HepG2, but not in mock transfected cell lines. VP3 could induce cell death by apoptosis in cancer cell lines, but not in diploid cell lines. All the results indicated that CAV vp3 gene, a potential therapeutic agents, has the potential of being used for cancer treatment.
Animals ; Apoptosis ; drug effects ; Capsid Proteins ; genetics ; Carcinoma, Hepatocellular ; pathology ; Cell Line, Tumor ; Chicken anemia virus ; Chickens ; Cloning, Molecular ; Eukaryotic Cells ; Genetic Vectors ; Humans ; Lipids ; Liver Neoplasms ; pathology ; Polymerase Chain Reaction ; Recombinant Proteins ; biosynthesis ; genetics ; pharmacology ; Sequence Analysis, DNA ; Transfection ; Viral Proteins ; genetics

Using PCR technique, the vp3 gene of chicken anemia virus (CAV) was cloned into the eukaryotic expression vector pcDNA3 to construct a recombinant pcDNA-vp3. Restriction enzyme digestion and sequencing analysis revealed that CAV vp3 gene was correctly inserted into the blank vector pcDNA3. After LipofectAMINE-mediated transfection in vitro with pcDNA-vp3 and pcDNA3 respectively, the total mRNA was extracted from liver carcinoma cell lines HepG2 and diploid cell line L-02, and RT-PCR was performed afterward. The results of RT-PCR suggested that vp3 gene was expressed in these two cell lines. At the same time, using in situ apoptotic detection assay, TUNEL kits, the apoptotic cells were found in pcDNA-vp3 transfected HepG2, but not in mock transfected cell lines. VP3 could induce cell death by apoptosis in cancer cell lines, but not in diploid cell lines. All the results indicated that CAV vp3 gene, a potential therapeutic agents, has the potential of being used for cancer treatment.
Apoptosis/*drug effects ; Capsid Proteins/*genetics ; Carcinoma, Hepatocellular/pathology ; Cell Line, Tumor ; Chicken anemia virus ; Chickens ; Cloning, Molecular ; Eukaryotic Cells ; Genetic Vectors ; Lipids ; Liver Neoplasms/*pathology ; Polymerase Chain Reaction ; Recombinant Proteins/biosynthesis ; Recombinant Proteins/genetics ; Recombinant Proteins/pharmacology ; Sequence Analysis, DNA ; *Transfection ; Viral Proteins/genetics

To observe the possibility of human visceral larva migrans due to eating of raw liver of domestic animals, especially of cattle, and also to serve as a good reference for adequate sanitary measures, the investigation survey was carried out from May 1975 to May 1976. From the subjects of a l,048 inhabitants (male 558, female 490) in five localities including two Provinces and three different cities, food habit was studied by questionnaire mannual. Larvae isolated from liver tissues of cattle, and pig were identified. Experimental observation on the chicken and mice infected with Toxocara canis was undertaken to draw a assumption of possibility inducing human visceral larva migrans. The results obtained from the present study are summarized. A part of Korean people has the habit to eat the livers of cattle, fowl, pig and dog raw. Eating rate of raw beef liver was 37.8 percent out of l,048 inhabitants, and its rate was higher markedly in male(57.7 percent) than in female (15. 1 percent), and the highest rate among the group of 31-40 years old. Eating rate of raw liver of fowl was 5.9 percent, pig 5.3 percent, and dog 2.5 percent. Larva recovery rate from beef liver was 11.8 percent out of 195 samples and 72.0 percent of total detected 1arvae were identified as Toxocara(=Neoascaris) vitulorum. From pig liver, larvae of nematoda were found in 6.4 percent out of 109 samples but no larva was detected from 120 fowl livers. Larvae detected from one-half of tissues and organs of infected chicken with about 2,000 Toxocara canis eggs were 8-245 in number, and 85-100 percent of recovered larvae were from their 1iver tissues. Toxocara canis larvae, 45, 31, 42 and 23 in number at 3rd, 14th, 25th and 55th day in one-half of the tissues and organs after infection respectively, were demonstrated from the mice infected with 500 larvae collected from infected chicken liver. Most of the larvae were recovered from the carcass of the mouse. It was approved the larvae isolated from chicken possess infectivity to the mice. Typical eosinophilic granulomatous change was not observed in the liver tissue of the infected chicken at 20th day after infection. As it summarized above, the liver of various domestic animals is the favorite tissue for migration of nematodes larvae. Therefore, the possibility of human visceral larva migrans may be induced due to eating of raw liver of domestic animals.
parasitology-helminth-nematoda ; visceral larva migrans ; Toxocara canis ; liver ; cattle ; fowl ; pig ; dog ; mouse ; chicken ; infectivity

An experimental study was carried out to observe the susceptibility of several kinds of laboratory animals to Fibricola seoulensis infection, a diplostomatid fluke of mammals. The metacercariae were obtained from the viscera of the snakes, Natrix tigrina lateralis and 50-2,000 in number each was artificially fed to a total of 127 animals; albino rats, mice, dogs, cats, rabbits and chickens. After 3 days to 8 weeks the animals were sacrificed and the recovery rate of worms as well as their maturity was observed. The results are as follows: The overall wom recovery rates throughout the experimental period was highest in albino rats(40.0 %) followed by mice(33.9%), cats(20.9 %), dogs(11.4 %), rabbits(0.05 %) and chickens(0 %). However, the recovery rates in the same host decreased as infection progressed longer and variable by the amount of metacercariae given. From albino rats and mice, the highest recovery rates were obtained in 1,000 and 200 metacercariae infection groups repectively, and it is considerd that such amount should be the optimum dose for experimental infection of these animals. The main location of F. seoulensis in experimental animals was small intestine especially the duodenum. The maturity index (No. mature worms/No. examined) was 100% in albino rats and mice, while only 22.7% or 0% in dogs or cats respectively. From the results, it is concluded that albino rats and mice are the most susceptible hosts for F. seoulensis infection among six kinds of laboratory animals examined.
parasitology-helminth-trematoda ; Fibricola seoulensis ; development ; biology ; albino rats ; mouse ; dog ; cat ; rabbit ; chicken

OBJECTIVE: To investigate the anti-tumor effects and the mechanism of the recombinant fowlpox virus expressing Apoptin gene on human laryngeal carcinoma Hep-2. METHOD: Hep-2 cells cultured in vitro were infected with vFVApoptin. The anti-tumor effects on Hep-2 cells were measured through MTT staining and, the mitochondrial trans-membrane potential (delta psi m) and reactive oxygen species (ROS) were analyzed by flow cytometry. Western blot was used to detect the release of cytochrome c (Cyto c). Caspase-3/9 activities were measured by colorimetric assay. RESULT: vFVApoptin could restrain Hep-2 cells significantly and, had the function of down-regulating delta psi m, up-regulating ROS, promoting Cyto c release and activating Caspase-3/9. CONCLUSION Cyto c were released from mitochondria by the function of up-regulating ROS of vFVApoptin. Cyto c triggered Caspase-9 and, after the activation of Caspase-9, downstream apoptotic factors, such as caspase-3, were activated. Eventually, Hep-2 cells were suppressed by mitochondrial pathway apoptosis induced by vFVApoptin.
Animals ; Apoptosis ; drug effects ; Capsid Proteins ; genetics ; pharmacology ; Chicken anemia virus ; genetics ; Fowlpox virus ; genetics ; Humans ; Tumor Cells, Cultured

Capsid Proteins ; genetics ; therapeutic use ; Chicken anemia virus ; genetics ; Gene Transfer Techniques ; Genes, Viral ; Hep G2 Cells ; Humans ; Laryngeal Neoplasms ; genetics