Porcine circovirus type 2 (PCV2) infection decreases the efficacy of an attenuated classical swine fever virus (CSFV) vaccine

ABSTRACT: The Lapinized Philippines Coronel (LPC) vaccine, an attenuated strain of classical swine fever virus (CSFV), is an important tool for the prevention and control of CSFV infection and is widely and routinely used in most CSF endemic areas, including Taiwan. The aim of this study was to investigate whether PCV2 infection affects the efficacy of the LPC vaccine. Eighteen 6-week-old, cesarean-derived and colostrum-deprived (CDCD), crossbred pigs were randomly assigned to four groups. A total of 105.3 TCID50 of PCV2 was experimentally inoculated into pigs through both intranasal and intramuscular routes at 0 days post-inoculation (dpi) followed by LPC vaccination 12 days later. All the animals were challenged with wild-type CSFV (ALD stain) at 27 dpi and euthanized at 45 dpi. Following CSFV challenge, the LPC-vaccinated pigs pre-inoculated with PCV2 showed transient fever, viremia, and viral shedding in the saliva and feces. The number of IgM+, CD4+CD8-CD25+, CD4+CD8+CD25+, and CD4-CD8+CD25+ lymphocyte subsets and the level of neutralizing antibodies against CSFV were significantly higher in the animals with LPC vaccination alone than in the pigs with PCV2 inoculation/LPC vaccination. In addition, PCV2-derived inhibition of the CSFV-specific cell proliferative response of peripheral blood mononuclear cells (PBMCs) was demonstrated in an ex vivo experiment. These findings indicate that PCV2 infection decreases the efficacy of the LPC vaccine. This PCV2-derived interference may not only allow the invasion of wild-type CSFV in pig farms but also increases the difficulty of CSF prevention and control in CSF endemic areas.     

Hydatidosis in a Chapman's zebra (Equus burchelli antiquorum).

A 14-year-old male Chapman's zebra had been treated for a cervical granulomatous cellulitis for 3 months prior to sudden death associated with myocardial fibrosis. Incidental findings at necropsy included multiple 3-8-cm unilocular cysts in the liver and lungs. Cysts contained either a clear fluid with or without hydatid sand or a gelatinous substance with foci of mineralization. Light and scanning electron microscopic examination of the hydatid sand revealed free protoscolices as well as intact and ruptured brood capsules with protoscolices attached to the germinal membrane. The protoscolices had 2 rows of 36-38 rostellar hooks with a length of 25-30 microm. The cyst wall consisted of inner germinal, intermediate laminated, and outer adventitial layers. Hydatidosis caused by Echinococcus granulosus was diagnosed based on the unilocular cysts, multiple protoscolices formed in a brood capsule, typical trilayered cyst wall, and herbivorous intermediate host. This is the first reported case of animal hydatidosis in Taiwan. The infection is thought to have been established 12 years ago in South Africa prior to importation.     

Vaccine-associated rhabdomyosarcoma with spinal epidural invasion and pulmonary metastasis in a cat

A 7-year-old, female, domestic medium-haired cat had a recurrent deep dermal mass in the interscapular region after initial surgical removal 3 months earlier. The cat had received a killed rabies vaccine and a five-in-one vaccine in the same area about 2 months prior to the first surgery. The relapsed mass was diagnosed as vaccine-associated sarcoma. The cat was euthanized 2 months later because of hind limb paralysis. At necropsy, multiple, poorly demarcated, nodular masses were seen in the muscles around the shoulders, neck, and thoracic vertebrae. Pulmonary metastasis and spinal epidural invasion at T1-T3 with regional cord compression and malacia were observed. Microscopically, the masses consisted of interwoven bundles of spindle cells with prominent multinucleated giant cell formation. The neoplastic cells stained strongly positive for myoglobin, and moderately but variably positive for vimentin, desmin, and alpha- smooth muscle actin. Phosphotungstic acid-hematoxylin staining revealed cytoplasmic striations in scattered tumor cells. The tumor was considered a vaccine-associated rhabdomyosarcoma.     

Coexisting tricuspid valve dysplasia and ventricular septal defect in a young patas monkey (Erythrocebus patas).

A 1.5-kg, 6-mo-old male patas monkey (Erythrocebus patas) was cyanotic and panting. Evaluation of the heart by electrocardiography, thoracic radiography, two-dimensional echocardiography, and Doppler color-flow echocardiography revealed a ventricular septal defect (VSD) with right-to-left shunting as well as tricuspid valve dysplasia with dilated annulus of the tricuspid ring, dilated right atrium, dilated right ventricle, and deformity of the tricuspid valve. Because of the severity of the cardiac disease, the patas monkey had complications recovering from anesthesia and died 3 days later. Gross postmortem findings included VSD, tricuspid dysplasia, and cerebral hemorrhage.     

Characterization of vitellogenin induced by estradiol-17β in protandrous black porgy,Acanthopagrus schlegeli

The objectives of this study were to isolate and characterize vitellogenin from plasma of estradiol-treated protandrous black porgy,Acanthopagrus schlegeli. Vitellogenin concentrations in plasma measured by a validated enzyme-linked immunosorbent assay (ELISA) were also compared. Two-year-old black porgy (n=20) were fed with estradiol-17 (4 mg kg^–1 of feed). Plasma was collected for purification of vitellogenin. Two forms of vitellogenin were found in plasma after chromatography on Sepharose CL-2B column and hydroxylapatite, and polyacrylamide gel electrophoresis. Black porgy vitellogenins are phospho-lipo-glycoproteins based on their chemical staining properties. The apparent molecular weights of the two forms of vitellogenin were 636 kDa and 321 kDa, respectively. The amino acid composition of purified vitellogenin was also analyzed after acid hydrolysis. The presence of immunoreactive vitellogenin was identified in the plasma and mucus extract from control and estradiol-induced females on the basis of Western blotting. Serial dilution of the plasma and mucus extract taken from estradiol-induced black porgy showed reactivity to an antiserum against lipovitellin in the ELISA, whereas mucus extract and plasma from male fish did not. Significantly higher concentrations of plasma vitellogenin were detected in estradiol-stimulated black porgy than in the control males.     

Development and evaluation of an indirect in situ polymerase chain reaction for the detection of porcine circovirus type 2 in formalin-fixed and paraffin-embedded tissue specimens

Taking advantage of the high sensitivity of polymerase chain reaction (PCR) and the cell-localizing ability of in situ hybridization (ISH), an indirect in situ PCR (ISPCR) method was developed for detecting the distribution of porcine circovirus type 2 (PCV2) in formalin-fixed and paraffin-embedded inguinal lymph nodes obtained from clinically healthy PCV2-carrier pigs and postweaning multisystemic wasting syndrome (PMWS)-affected pigs. Comparisons of the relative sensitivity of indirect ISPCR with other routinely used diagnostic methods for PCV2 indicated that nested PCR was the most sensitive method followed by indirect ISPCR, conventional PCR, ISH, and immunohistochemical (IHC) staining. Although indirect ISPCR, ISH, and IHC staining all revealed a similar signal distribution pattern of PCV2, using indirect ISPCR allowed specific amplification and detection of previously uneasily detected PCV2 signal than by routine ISH or IHC staining, particularly in those cells within the germinal center in clinically healthy PCV2-carrier pigs. Furthermore, six different PCV2 signal expression patterns in conjunction with the correlated lymphoid lesion stages were classified to describe the tissue morphological changes and viral infection. The result indicates that indirect ISPCR is a more effective, cell-based diagnostic tool with good specificity to detect limited PCV2 infection in formalin-fixed and paraffin-embedded tissue specimens and it would be a useful tool for further exploring the pathogenesis of PCV2 infection.     

The fatty acid composition and its alterations due to different incubation temperature in the lipids of aggressive and non-aggressive strains of Ceratocystis ulmi

Lipids and their fatty acids from the mycelia of aggressive and non-aggressive strains of Ceratocystis ulmi were examined at different incubation temperature. The quantity and degree of fatty acid unsaturation were correlated with the pathogenicity of the fungus.     

Ultrastructure of Ceratocystis ulmi

Transmission electron microscopic observations of C. ulmi show that there are three distinct types of the hyphae and three distinct layers forming the nscocarp peridium. The cellular organelles of the hyphae and ascocarp peridial layers are described.