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PubMed2005

Porcine Toll-like receptor 1, 6, and 10 genes: complete sequencing of genomic region and expression analysis.

Toll-like receptors (TLRs) recognize various microbial components and play key roles in activating the innate immune system. Hence, their function is important in swine infectious diseases. We completely determined 173,804 bp of nucleotide sequence of a genomic region including porcine TLR6 and the newly identified porcine TLR homologues TLR1 and TLR10. The porcine genomic structure of these genes was highly conserved in comparison with the corresponding region in humans. Analysis of their expression in porcine tissues showed differences in expression patterns between porcine TLR10 and TLR1 or TLR6. Moreover, phylogenetic analysis of the cytoplasmic regions of TLR genes suggested that the signal transduction pathway of TLR10 was different from those of TLR1 and TLR6. We also developed six polymorphic microsatellite markers within this genomic region; these markers will be valuable for association studies between TLR genes and resistance or susceptibility to infectious diseases in swine.

Amino Acid Sequence↗

PubMed2005

Veterinary public health activities at FAO: cysticercosis and echinococcosis.

In many developing and transition countries, parasitic zoonoses such as cysticercosis and echinococcosis cause serious human suffering and considerable losses in agricultural and human productivity, thus posing a significant hindrance to their development. Although, effective and reliable tools for the diagnosis, prevention and control of parasitic zoonoses are now available, their implementation has not always been successful in many countries. This is primarily due to the lack of awareness on the presence or impact of the causing parasites (Taenia saginata, Taenia solium and Echinococcus spp.). In addition, often the needed intersectorial cooperation, resource management and political commitment for their control are (also) absent. FAO's regular programme has established a global network of professionals directly involved in zoonotic and food-borne diseases. The network provides a basic framework for the spread of information related to the diagnosis, prevention and control of major zoonotic diseases including cysticercosis and echinococcosis.

Animals↗

PubMed2004

Screening and analysis of porcine endogenous retrovirus in Chinese Banna minipig inbred line.

Pigs have been the most likely animal as the source of cells, tissues, and organs for xenotransplantation. But the use of pigs in xenotransplantation is associated with the risk of porcine endogenous retrovirus (PERV) transmission. Previous studies have identified that the proviruses are integrated into the genome of normal pigs and that virus particles released from the porcine cells can infect human cells in vitro. As a unique inbred pig, Banna minipig inbred (BMI) has a huge potential value for xenotransplantation and medical research. It has been the focal experimental animal for pig-to-human xenotransplantation in China, due to its clear genetic background and tiny individual differences. To evaluate whether the potential risk of PERV exists in inbred pigs, a series of screening experiments were performed herein. The results of PCR with primers specific for gag, pol, and env showed that proviruses existed in the genome of BMI, and the PERV subtypes were PERV-A and PERV-B. PERV mRNA was expressed functionally in BMI. Positive results of an RT assay identified that PERV in BMI had potential infectivity, but the concentration of PERV reverse transcriptase in BMI was almost 20 times lower than that of HIV. These results suggested that gag, pol and env genes of PERV were not lost during inbreeding, which created favorable conditions to produce viral particles that could possibly infect human cells in xenotransplantation.

Animals↗

PubMed2006

Concurrent swine erysipelas and Clostridium novyi infections associated with sow mortality in outdoor sows in Kenya.

A sudden increase in sow mortality was reported from a large outdoor pig-breeding unit in Kenya. The sows showed pyrexia and diamond shaped to confluent skin lesions ante-mortem. Gross pathological examination of the carcases was performed and organ samples (heart, liver, kidney, spleen, mandible lymph nodes) were sent for further diagnostic investigation. Based upon the clinical, gross- and microscopic-pathological signs, bacteriological findings and toxin testing, concurrent infection with Erysipelothrix rhusiopathiae and Clostridium novyi was diagnosed as the cause of the sow mortality.

Animals↗

PubMed2004

Differentiating infection from vaccination in foot-and-mouth-disease: evaluation of an ELISA based on recombinant 3ABC.

Recent devastating outbreaks of foot-and-mouth disease (FMD) in Europe have reopened the discussion about the adequacy of the non-vaccination strategy implemented by the EU in 1991. Here we describe the evaluation of a new commercially available test kit for the discrimination between vaccination and infection. The test is based on the detection of antibodies against the recombinant non-structural (NS) protein 3ABC. In contrast to immunization with vaccines free of 3ABC, these antibodies are elicited as a consequence of infection. Testing more than 3600 negative sera from several countries revealed a specificity of > 99% for bovine, ovine, and porcine samples. Antibodies specific for 3ABC can be detected as soon as 10 days post-infection. As compared with the occurrence of antibodies against structural proteins of FMDV, anti-3ABC antibodies can be detected 5-10 days later, depending on the species. No anti-3ABC antibodies were detected in sera from vaccination experiments or in field sera from vaccinated animals. However, anti-3ABC antibodies can be detected in vaccinated animals upon challenge. These results provide evidence that this test can facilitate the use of vaccines in new strategies against FMD.

Animals↗

PubMed2004

Procedures for preventing transmission of foot-and-mouth disease virus (O/TAW/97) by people.

The aim of this study was to determine personal hygiene protocols and animal avoidance periods needed to prevent transmission of FMDV (O/TAW/97). Forty-six, 9-week-old barrows free of FMDV were randomly allocated to five treatment groups and a control group. Investigators contacted and sampled FMDV-inoculated pigs for approximately 40 min and then contacted and sampled sentinel pigs after using no biosecurity procedures, washing hands and donning clean outerwear, or showering and donning clean outerwear. Personnel were sampled for nasal carriage of FMDV for 85.43 h. Contaminated personnel did not transmit FMDV to susceptible pigs after handwashing or showering, and donning clean outerwear. FMDV was transmitted when biosecurity procedures were not used. FMDV was not detected in nasal secretions of investigators. Thus, extended animal avoidance periods do not appear to be necessary to prevent transmission of FMDV (O/TAW/97) by people to pigs when organic material is removed through handwashing/showering and donning clean outerwear. This study supports similar findings in a previous publication using FMDV (O/UK/35/2001).

Animal Husbandry↗

PubMed2005

Immunogenicity of a recombinant pseudorabies virus expressing ORF1-ORF2 fusion protein of porcine circovirus type 2.

Porcine circovirus type 2 (PCV2) is associated with post-weaning multisystemic wasting syndrome (PMWS). Pseudorabies (PR) is also an important infectious disease in swine and sometimes co-infect with PCV2. An attenuated pseudorabies virus (PRV) has been successfully used as a vector for live viral vaccines. In this study, a recombinant PRV expressing ORF1-ORF2 fusion protein of PCV2 was constructed and its immunogenicity was tested in mice and pigs. The ORF1 and partial ORF2 gene of PCV2 Yu-A strain were amplified by PCR and inserted into a transfer vector. The recombinant transfer plasmid was co-transfected with the EcoRI digested genome of vector virus (PRV TK-/gE-/LacZ+) into IBRS-2 cells. The recombinant pseudorabies virus PRV-PCV2 was purified by plaque purification and identified by PCR and Southern blotting. Expression of the ORF1-ORF2 fusion protein by the recombinant PRV-PCV2 virus was demonstrated by Western blotting analysis. The growth properties of the recombinant virus in cells were similar to that of the parent vector virus. In animal experiments, PRV-PCV2 elicited strong anti-PRV and anti-PCV2 antibodies in Balb/c mice as indicated by PRV-neutralizing assay, anti-PCV2 ELISA and PCV2 specific lymphocyte proliferation assay, respectively. And PRV-PCV2 immunization protected mice against a lethal challenge of a virulent PRV Ea strain. In pigs, PRV-PCV2 elicited significant immune response towards PRV and PCV2 as indicated by PRV-ELISA, PRV neutralizing assay and PCV2 specific lymphocyte proliferation assay, respectively. This is a first step toward the development of a potential candidate divalent vaccine against PRV and PCV2 infections.

Animals↗

PubMed2006

Inhibition of porcine circovirus type 2 replication in mice by RNA interference.

Porcine circovirus type 2 (PCV2) is the primary causative agent of an emerging swine disease, postweaning multisystemic wasting syndrome (PMWS) for which no antiviral treatment is available. To exploit the possibility of using RNA interference (RNAi) as a therapeutic approach against the disease, plasmid-borne short hairpin RNAs (shRNAs) were generated to target the PCV2 genome. Transfection of these shRNAs into cultured PK15 cells caused a significant reduction in viral RNA production that was accompanied by inhibiting viral DNA replication and protein synthesis in infected cells. The effect was further tested in vivo in a mouse model that has been developed for PCV2 infection. Mice injected with shRNA before PCV2 infection showed substantially decreased microscopic lesions in inguinal lymph nodes compared to controls. In situ hybridization and immunohistochemical analyses showed that shRNA caused a significant inhibition in the level of viral DNA and protein synthesis detected in the lymph nodes of the treated mice relative to the controls. Taken together, these results indicate that shRNAs are capable of inhibiting PCV2 infection in vitro as well as in vivo and thus may constitute an effective therapeutic strategy for PCV2 infection.

Animals↗

PubMed1999

Characterization of porcine and ovine Oesophagostomum spp. by isoenzymatic patterns and restriction-fragment-length polymorphisms (RFLPs).

Four different morphological and biometrical populations of Oesophagostomum have been identified, using classical taxonomy methods, from Sus scrofa domestica (pigs): O. dentatum (Od), O. quadrispinulatum (Oq), O. granatensis (Og), and a fourth population, including individuals with morphological and biometric parameters overlapping these three species that were clasified as Oesophagostomum sp. The G6PD and MDH isoenzymatic patterns did not discriminate between the three species, while GPI showed a diagnostic isoenzymatic pattern for Oq. Og showed identical G6PD, GPI and MDH isoenzymatic pattern as Od. Furthermore, after rDNA amplification by polymerase chain reaction (PCR), the uncut PCR product showed that the ITS2 of these three species had a similar size of 320 base pairs (bp). Restriction-fragment-length polymorphisms (RFLP) were analyzed after digestion of the ITS2 with 13 different restriction enzymes. After electrophoretic separation of the digested PCR products, only one unique differentiating pattern of bands was observed for Od and Oq. This was when Sau3AI was used, while Og showed an identical band pattern to Od. Thus, our studies provided no evidence for the existence of Og and Od as differentiated populations. O. venulosum was isolated from sheep and goat; G6PD and MDH isoenzymatic patterns discriminated this species from porcine species of Oesophagostomum. The ITS2 region appeared as a different band of 380 bp from those observed for porcine Oesophagostomum species.

Animals↗

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