Development of an indirect ELISA for serological diagnosis of bovine herpesvirus 5

Bovine herpesviruses 1 and 5 (BoHV-1 and BoHV-5) are economically important pathogens, associated with a variety of clinical syndromes, including respiratory and genital disease, reproductive failure and meningoencephalitis. The standard serological assay to diagnose BoHV-1 and BoHV-5 infections is the virus neutralization test (VNT), a time consuming procedure that requires manipulation of infectious virus. In the present study a highly sensitive and specific single dilution indirect ELISA was developed using recombinant glycoprotein D from BoHV-5 as antigen (rgD5ELISA). Bovine serum samples (n = 450) were screened by VNT against BoHV-5a and by rgD5ELISA. Compared with the VNT, the rgD5ELISA demonstrated accuracy of 99.8%, with 100% sensitivity, 96.7% specificity and coefficient of agreement between the tests of 0.954. The rgD5ELISA described here shows excellent agreement with the VNT and is shown to be a simple, convenient, specific and highly sensitive virus-free assay for detection of serum antibodies to BoHV-5

Cloning and expression of a chimera containing ROP2 of Neospora caninum fused with OprI lipoprotein from Pseudomonas aeruginosa / Clonagem e expressão de quimera contendo a proteína ROP2 de Neospora caninum fusionada a lipoproteína OprI de Pseudomonas aeruginosa

Neospora caninum is the etiologic agent of neosporosis, is one of the main responsible for abortion in cattle herds, causing economic losses to Cattle-raising. Vaccination of cattle would be an important alternative; however, the lack of effective vaccines prevents the application of this control method. The proteins present in rhoptries (ROPs), due to their importance in cell infection and their antigenic and immunogenic characteristics, became excellent candidates for vaccine antigens. The bacterial lipoproteins as an Oprl from Pseudomonas aeruginosa have received particular attention as an adjuvant carrier molecule. The aims of this study were to clone and expressing a chimera containing NcROP2 fused with OprI lipoprotein from P. aeruginosa for the future development of a recombinant vaccine against N. caninum. We cloned and expressed, in Escherichia coli Rosetta (DE3) pLysS, the region of NcROP2 described between amino acids 191 and 359, fused OprI producing the chimera rROP2/OprI, showed an expected size of ~50 kDa. and characterized its antigenicity. The protein was purified and characterized by Western blot with anti-histidine monoclonal antibodies and their antigenicity recognized by sera from animals naturally infected by N. caninum. The chimera rROP2/OprI was recognized by antibodies anti-N. caninum reviling common antigenic determinants of the recombinant protein and its native form, suggesting its use for developing a recombinant vaccine

Origanum vulgare (Lamiaceae) OVICIDAL POTENTIAL ON GASTROINTESTINAL NEMATODES OF CATTLE

Due to anthelmintic resistance in nematodes, several research studies have been developed seeking control alternatives to these parasites. This study evaluated the in vitro action of Origanum vulgare on gastrointestinal nematode eggs of cattle. In order to evaluate the ability to inhibit egg hatch, different dried leaves extracts of this plant were tested, such as dye, hydroalcoholic and aqueous extracts at concentrations varying from 0.62 to 80 mg/mL. Each assay was accompanied by control containing levamisole hydrochloride (0.2 mg/mL), distilled water and 70 ºGL grain alcohol at the same concentration of the extracts. Test results showed that the different O. vulgare extracts inhibited egg hatch of cattle gastrointestinal nematodes at a percentage that varied from 8.8 to 100%; dye and hydroalcoholic extract were the most promising inhibitors. In view of this ovicidal property, O. vulgare may be an important source of viable antiparasitic compounds for nematodiosis control in ruminants

Efeito imunomodulador do bacillus toyonensis em equinos vacinados contra o tétano

Este estudo objetiva avaliar a modulação da resposta imunológica em equinos suplementados com o probiótico B. toyonensis vacinados com vacina recombinante contra a toxina de Clostridium tetani

Bovine herpesvirus glycoprotein D: a review of its structural characteristics and applications in vaccinology

International audienceThe viral envelope glycoprotein D from bovine herpesviruses 1 and 5 (BoHV-1 and -5), two important pathogens of cattle, is a major component of the virion and plays a critical role in the pathogenesis of herpesviruses. Glycoprotein D is essential for virus penetration into permissive cells and thus is a major target for virus neutralizing antibodies during infection. In view of its role in the induction of protective immunity, gD has been tested in new vaccine development strategies against both viruses. Subunit, DNA and vectored vaccine candidates have been developed using this glycoprotein as the primary antigen, demonstrating that gD has the capacity to induce robust virus neutralizing antibodies and strong cell-mediated immune responses, as well as protection from clinical symptoms, in target species. This review highlights the structural and functional characteristics of BoHV-1, BoHV-5 and where appropriate, Human herpesvirus gD, as well as its role in viral entry and interactions with host cell receptors. Furthermore, the interactions of gD with the host immune system are discussed. Finally, the application of this glycoprotein in new vaccine design is reviewed, taking its structural and functional characteristics into consideration