Serodiagnostic comparison of enzyme-linked immunosorbent assay and surface plasmon resonance for the detection of antibody to Porcine circovirus type 2

This paper describes the cloning and expression of the capsid protein of Porcine circovirus type 2 (PCV2) in an Escherichia coli expression system that was used to produce a fusion protein for subsequent immunologic studies: enzyme-linked immunosorbent assay (ELISA) and surface plasmon resonance (SPR). Polymerase chain reaction was used to amplify the gene encoding the capsid protein from the DNA of PCV2. The protein was then cloned into a pRSET prokaryotic expression vector. Western blot analysis revealed that the recombinant protein gave strong signals on a polyvinylidene difluoride membrane when exposed to the serum from a pig infected with PCV2. The expressed protein was purified and used as an antigen for the ELISA and SPR study. A protein chip based on SPR was developed, and the diagnostic potential of SPR was compared with that of ELISA with the use of 70 serum samples obtained from 6 pig farms. There was a strong positive correlation between the ELISA and SPR titers (r = 0.877, P < 0.01). Therefore, this recombinant capsid protein can be used as an antigen for serologic studies, and the SPR, a label-free method, appears to be a valuable and reproducible tool in the serodiagnosis of a PCV2 infection.     

Increased humoral antibody response of foot-and-mouth disease virus vaccine in growing pigs pre-treated with poly-γ-glutamic acid

This study was conducted to determine if humoral antibody response of foot-and-mouth disease (FMD) vaccine improved in 8-week-old growing pigs born to well-vaccinated sows pre-treated with 60 mg of poly-γ-glutamic acid (γ-PGA) three days before vaccination. Antibody against FMD virus serotype O was measured 0, 2, 4 and 6 weeks post-vaccination, using a PrioCHECK FMDV type O ELISA kit. The results showed that positive antibody reactions against FMDV serotype O antigen among a component of the vaccine significantly increased in response to pre-injection with γ-PGA.     

Comparison of antibody titers in rabbits following immunization with inactivated influenza virus via subarachnoidal or subcutaneous route

Rabbits were immunized with inactivated influenza virus via the subarachnoidal (SA) or subcutaneous (SC) route, and the antibody titers in cerebrospinal fluid (CSF) and serum were assayed. There were no nervous signs or morphological lesions related to SA immunization. In the SC group, the antibody titer was elevated in serum, but not elevated in CSF. In the SA group, the antibody titer was significantly elevated in serum and even in CSF, and their antibody titers were much greater than in the SC group. The present results suggest that intrathecal immunization is more effective than SC immunization at inducing a protective immune response against the transneural spread of viruses.     

Gingival overgrowth in dogs associated with clinically relevant cyclosporine blood levels: observations in a canine renal transplantation model

Objective— To investigate the development of gingival hyperplasia in dogs after renal transplantation and administration of microemulsified cyclosporine A (MCsA).
Study Design— Experimental study.
Animals— Healthy adult mongrel dogs (n=5).
Methods— As part of study on renal transplantation, dogs administered MCsA (20 mg/kg/day), azathioprine, and prednisolone to prevent graft rejection were monitored for development of gingival changes. Prednisolone was discontinued after 3 months. MCsA dose was adjusted to maintain whole blood trough concentration of 400–700 ng/mL. Gingival change was evaluated by weekly examination and photodocumentation, and gingival biopsy for histopathology was performed at 28 weeks.
Results— One dog was lost because of acute graft rejection. Gingival hyperplasia developed in 3 of 4 dogs. The earliest gingival changes occurred in the interdental papillae at 20 weeks after transplantation. On histopathology, the underlying connective tissue was thickened and contained increase numbers of fibroblasts and inflammatory infiltrates.
Conclusions— Long-term immunosuppression with an MCsA-based treatment likely induces substantial gingival hyperplasia when therapeutic, immunosuppressive blood levels of MCsA were maintained for 32 weeks.
Clinical Relevance— MCsA is used for immune-mediated diseases and preventing rejection after transplant in dogs. MCsA blood levels, and gingival hyperplasia should be monitored by routine examination of the interdental papilla in dogs administered MCsA for long periods.     

Effects of chemical treatments on pH and bacterial population in poultry litter: a laboratory experiment

1. The goal of this study was to evaluate the effects of three chemical treatments on pH and bacterial populations (total aerobic bacteria and gram-negative bacteria) in poultry litter under laboratory conditions. 2. Litter obtained from poultry houses was treated with three chemical treatments (alum, AlCl(3) and FeSO(4)) at the same concentration (8 g/100 g litter), while untreated litter served as a control. The study was conducted for 3 weeks. 3. All of the chemical treatments reduced total aerobic bacteria (22 to 87% of the untreated control) and gram-negative bacteria (63 to 99% of the untreated controls) populations and lowered litter pH values (5.95 to 6.64). However, a significant difference in gram-negative bacteria did not exist among chemical treatments at 0, 1 and 2 weeks. 4. These results suggest that the reduction in total aerobic bacteria and gram-negative bacteria populations is highly related to a decrease in litter pH, and acidifying treatment (alum, AlCl(3) and FeSO(4)) of poultry litter may serve as a means to help the reduction in pathogen populations and to improve economical benefits under commercial production conditions.