The objectives of the present study were to evaluate the anatomic localization of porcine reproductive and respiratory syndrome virus (PRRSV) in naturally infected pigs and to determine whether oral fluid could be used to detect the virus in infected animals. Two sows, seven 2-month-old grower pigs, and 70 6-month-old gilts were included in this study. PRRSV in sera and oral fluid were identified by nested reverse transcription PCR (nRT-PCR) while lung, tonsil, and tissue associated with oral cavity were subjected to nRT-PCR, immunohistochemistry, and in situ hybridization. In sows, PRRSV was identified in oral fluid and tonsils. PRRSV was also detected in oral fluid, tonsils, salivary glands, oral mucosa, and lungs of all seven grower pigs. However, viremia was observed in only two grower pigs. Double staining revealed that PRRSV was distributed in macrophages within and adjacent to the tonsillar crypt epithelium. In gilts, the North American type PRRSV field strain was detected 3 to 8 weeks after introducing these animals onto the farm. These results confirm previous findings that PRRSV primarily replicates in tonsils and is then shed into oral fluid. Therefore, oral fluid sampling may be effective for the surveillance of PRRSV in breeding herds.
Animals ; Female ; In Situ Hybridization/veterinary ; Lung/virology ; Male ; Palatine Tonsil/virology ; Polymerase Chain Reaction/veterinary ; Porcine Reproductive and Respiratory Syndrome/*virology ; Porcine respiratory and reproductive syndrome virus/*physiology ; Saliva/*virology ; Salivary Glands/virology ; Swine/virology ; Virus Replication/physiology

OBJECTIVE: To investigate the epidemiology of EBV in adenoidal hypertrophy and chronic tonsillitis and discuss the affection of EBV on the nosogenesis of adenoidal hypertrophy and tonsillitis of children. METHOD: Fifty-two children with chronic tonsillitis and/or adenoidal hypertrophy had the operations of the tonsillectomy and/or the adenoidectomy. These tissues resected and plasma of all cases were detected to find EBV-DNA by RQ PCR. RESULT: The infection rate of EBV in the tissues of adenoidal hypertrophy and tonsillitis of children was 51.9%. The boys' infection rate of EBV was 50.0%, and the girls' infection rate of EBV was 55.6%, which had not significantly different. The EBV infection rate in the tissues of tonsillitis was 40.4%, The EBV infection rate in the tissues of adenoidal hypertrophy was 48.9%, which had not significant difference. The school age group (7- to 14-years-old) presented higher infection rate of EBV in the tissues of adenoid and tonsil (65.5%) than the pre-school children group (2- to 6-years-old) (34.8%). Comparing the copies numbers of EBV-DNA in the different degrees of adenoidal hypertrophy, we found that the copies numbers of EBV-DNA in the severe hypertrophy group were higher than the midrange and slight hypertrophy groups (P<0.05). Meanwhile we detected EBV-DNA in these childrens' blood plasma by RQ-PCR. No blood plasma was detected EBV-DNA copies higher than normal (< 1 x 10(3) copies/ml). CONCLUSION The tissues of adenoidal hypertrophy and tonsillitis had same sensitivity to EBV. There was not significant difference between the infection rates of the boys and girls with adenoidal hypertrophy and/or tonsillitis. With these children growing up and the course of diseases prolonging, the infection rate of EBV increased correspondingly. There was a certain correlation between the hypertrophy of adenoid and EBV. There were no EBV-DNA fragments in blood plasma of the children with adenoidal hypertrophy and/or tonsillitis. So there were essential different between benign hyperplasia and nasopharyngeal carcinoma.
Adenoids ; pathology ; virology ; Adolescent ; Child ; Child, Preschool ; DNA, Viral ; isolation & purification ; Epstein-Barr Virus Infections ; pathology ; Female ; Herpesvirus 4, Human ; Humans ; Hypertrophy ; pathology ; virology ; Male ; Palatine Tonsil ; pathology ; virology ; Tonsillitis ; pathology ; virology

Porcine circovirus (PCV) type2 was isolated using primary porcine kidney cells from lymph node of piglets with typical PMWS. The presence of the virus was identified by PCR using primers specific to PCV type2. The ORFs 1 and 2 were amplified by PCR using primers corresponding to the target genes of the PCV type 2. Cloned genes were inserted into the baculovirus expression vector and PCV recombinant proteins were expressed using baculovirus expression system. Recombinant protein expression was determined by indirect immunofluorescent assay (IFA) and immunoblotting using polyclonal antiserum to PCV. ORF1 gene expressed two proteins with approximately 17 kDa and 31 kDa proteins in the baculovirus system. Recombinant protein of the ORF2 was similar to that of the native virus except minor bands with different molecular weight were detected. Recombinant protein expressed in the baculovirus system showed at least two glycosylation sites based on the tunicamycin treatment. Recombinant protein of the ORF2 assembled virus-like particle in recombinant virus infected insect cells.
Animals ; Baculoviridae/*genetics ; Blotting, Western ; Circoviridae Infections/*veterinary/*virology ; Circovirus/*classification/genetics/isolation & purification/ultrastructure ; Cloning, Molecular ; Fluorescent Antibody Technique, Indirect ; Lymph Nodes/virology ; Microscopy, Electron ; Open Reading Frames ; Palatine Tonsil/virology ; Polymerase Chain Reaction/methods/veterinary ; Recombinant Proteins/analysis ; Swine ; Swine Diseases/*virology ; Transfection ; Tunicamycin/pharmacology ; Viral Proteins/*analysis