Preliminary binary ethylenimine (BEI) inactivated bovine papillomavirus (BPV) vaccine trial against cutaneous warts in bull calves: a pathological assessment

A preliminary prophylactic and therapeutic vaccine trial was successfully conducted in bull calves with binary ethylenimine (BEI) inactivated saponized crude cutaneous warts bovine papillomavirus 1 and 2 (CW BPV-1 & -2) vaccine. In Prophylactic Group of vaccinated animals no temperature, untoward reaction or nodule formation was seen at vaccine inoculation site. After challenge with BPV inoculum, cutaneous warts (CWs) failed to develop in this group indicating that vaccine was effective. In Therapeutic Group, before vaccination in two animals CWs were experimentally induced. In one animal, luxuriant cauliflower-like and in other linear slow growth was observed. After vaccination, CWs in both animals became dried and growth was arrested. It appeared that tumours were regressing which was confirmed by further clinical observations as very little tumour-scar was left at termination of experiment.Histopathologically, both cases were diagnosed as regressing occult/ early fibroblastic type papilloma. Mononuclear cellular infiltration/ aggregates around hair follicles and in tumour stroma were noted. As compared to first case (cauliflower- like/ fast growing) in second case (linear growth/ fast regressing), in addition to other findings mononuclear cellular inflammatory reaction were more pronounced. The clinical regression results are in accordance with histopathological findings of degree of infiltration of lymphocytes and this trial showed that both type use of BEI inactivated saponized crude CW BPV-1&-2 vaccine gave encouraging results with fast regression/ prevention of CWs.

Preliminary binary ethylenimine (BEI) inactivated bovine papillomavirus (BPV) vaccine trial against cutaneous warts in bull calves: a pathological assessment

A preliminary prophylactic and therapeutic vaccine trial was successfully conducted in bull calves with binary ethylenimine (BEI) inactivated saponized crude cutaneous warts bovine papillomavirus 1 and 2 (CW BPV-1 & -2) vaccine. In Prophylactic Group of vaccinated animals no temperature, untoward reaction or nodule formation was seen at vaccine inoculation site. After challenge with BPV inoculum, cutaneous warts (CWs) failed to develop in this group indicating that vaccine was effective. In Therapeutic Group, before vaccination in two animals CWs were experimentally induced. In one animal, luxuriant cauliflower-like and in other linear slow growth was observed. After vaccination, CWs in both animals became dried and growth was arrested. It appeared that tumours were regressing which was confirmed by further clinical observations as very little tumour-scar was left at termination of experiment.Histopathologically, both cases were diagnosed as regressing occult/ early fibroblastic type papilloma. Mononuclear cellular infiltration/ aggregates around hair follicles and in tumour stroma were noted. As compared to first case (cauliflower- like/ fast growing) in second case (linear growth/ fast regressing), in addition to other findings mononuclear cellular inflammatory reaction were more pronounced. The clinical regression results are in accordance with histopathological findings of degree of infiltration of lymphocytes and this trial showed that both type use of BEI inactivated saponized crude CW BPV-1&-2 vaccine gave encouraging results with fast regression/ prevention of CWs.(AU)

Immunogenicity of an inactivated bovine herpesvirus type 5 strain defective in thymidine kinase and glycoprotein E / Imunogenicidade de uma cepa inativada do herpesvírus bovino tipo 5 defectiva na timidina quinase e glicoproteína E

The immunogenicity of an inactivated, experimental vaccine based on a bovine herpesvirus type 5 strain defective in thymidine kinase and glycoprotein E (BoHV-5 gE/TKΔ) was evaluated in cattle and the results were compared with a vaccine containing the parental BoHV-5 strain (SV507/99). To formulate the vaccines, each virus (wildtype SV507/99 and BoHV-5 gE/TK∆) was multiplied in cell culture and inactivated with binary ethyleneimine (BEI). Each vaccine dose contained approximately of 107.5 TCID50 of inactivated virus mixed with an oil-based adjuvant (46:54). Forty calves, 6 to 9-months-old, were allocated into two groups of 20 animals each and vaccinated twice (days 0 and 22pv) by the subcutaneous route with either vaccine. Serum samples collected at day 0 and at different intervals after vaccination were tested for virus neutralizing (VN) antibodies against the parental virus and against heterologous BoHV-5 and BoHV-1 isolates. The VN assays demonstrated seroconversion to the respective homologous viruses in all vaccinated animals after the second vaccine dose (mean titers of 17.5 for the wildtype vaccine; 24.1 for the recombinant virus). All animals remained reagents up to day 116 pv, yet showing a gradual reduction in VN titers. Animals from both vaccine groups reacted in similar VN titers to different BoHV-1 and BoHV-5 isolates, yet the magnitude of serological response of both groups was higher against BoHV-5 field isolates. Calves vaccinated with the recombinant virus did not develop antibodies to gE as verified by negative results in a gE-specific ELISA, what would allow serological differentiation from naturally infected animals. Taken together, these results indicate that inactivated antigens of BoHV-5 gE/TK recombinant virus induced an adequate serological response against BoHV-5 and BoHV-1 and thus can be used as an alternative, differential vaccine candidate.(AU)

A imunogenicidade de vacina experimental inativada, produzida com uma cepa do herpesvírus bovino tipo 5 defectiva nos genes da timidina quinase e glicoproteína E (BoHV-5 gE/TKΔ) foi avaliada em bovinos e o resultado foi comparado com a resposta induzida pela cepa parental do BoHV-5 (SV507/99). Para a formulação da vacina, cultivos de células infectados com cada um dos vírus (SV507/99 ou BoHV-5 gE/TKΔ) foram inativados com etilenamina binária. Cada dose de vacina continha aproximadamente 107,5 TCID50 de um dos vírus inativados emulsionado em adjuvante oleoso (46:54). Quarenta bezerros de raças cruzadas, com idade entre seis a nove meses, foram alocados em dois grupos de 20 animais cada e vacinados duas vezes (dia 0 e 22 pv) pela via subcutânea com uma das vacinas. Amostras de soro foram coletadas no dia 0 e a vários intervalos após vacinação para a pesquisa de anticorpos neutralizantes frente ao vírus homólogo ou frente a isolados de BoHV-5 e BoHV-1. Os testes de soroneutralização (SN) demonstraram que todos os animais soroconverteram após a segunda dose da vacina (títulos médios de 17,5 para o grupo SV507/99; 24,1 para o grupo BoHV-5 gE/TKΔ). Todos os animais mantiveram níveis de anticorpos neutralizantes até o dia 116 pv, no entanto foi observada uma redução gradual no títulos. A sorologia cruzada com amostras heterólogas do BoHV-5 e BoHV-1 indicou que ambos os grupos vacinais reagiram em níveis similares frente ao mesmo vírus, no entanto a magnitude da resposta sorológica foi maior frente a amostras de BoHV-5. Os animais vacinados com a cepa recombinante não desenvolveram anticorpos contra a gE detectáveis por um ELISA específico, o que permitiria a sua diferenciação sorológica de animais infectados naturalmente. Esses resultados demonstram que a vacina contendo antígenos inativados do vírus recombinante BoHV-5 gE/TKΔ induziu resposta sorológica em níveis satisfatórios, constituindo-se, assim, em alternativa a cepa vacinal diferencial.(AU)

Entomopathogenic fungi isolates to control the borer of yerba mate (Hedypathes betulinus) Kluger (Coleoptera; Cerambycidae) / Isolados de fungos entomopatogênicos visando ao controle da broca da erva-mate (Hedypathes betulinus) Kluger (Coleoptera; Cerambycidae)

Yerba mate is an important crop of southern Brazil, Argentina and Paraguay, with socio-economic and environmental importance. Due the lack of native herbals, yerba mate has become a monoculture, and as a consequence, insects have become pests, and the borer, Hedypathes betulinus, one of the most important. In order to improve biological control in the crop, this work aims to select in laboratory the most virulent, of 32 isolates of the entomopathogenic fungi Beauveria bassiana (Bals.) Vuill. and 18 isolates of Metarhizium anisopliae (Metsch.) Sorok. Initially, isolates virolence was evaluated by immersing the adults borers in a fungal suspension, containing 1×108 conidia mL-1, and after, those caused highest confirmed mortality were selected for the second step, wherein it was evaluated the virulence, vegetative growth and conidia production in culture medium, rice and borer bodies. Three isolates of B. bassiana (UNIOESTE 4, UNIOESTE 52 and UNIOESTE 64) and one of M. anisopliae (IBCB 352) reached confirmed mortality equal or superior to 90%, these isolates were selected for the second step, together with the standard isolated GC 716 (B. bassiana). The isolated IBCB 352 of M. anisopliae, despite being the only of this species to be selected for the second step, promoted the high confirmed mortality and conidiogenesis in adults borer, indicating a high inoculums production i, bei

A erva-mate é uma cultura importante da região sul do Brasil, Argentina e Paraguai, tendo grande importância sócio-econômica e ambiental. Com a escassez de ervais nativos, a cultura da erva-mate passou a ser cultivada em sistema de monocultivo e como consequência, alguns insetos tornaram-se pragas, sendo a broca, Hedypathes betulinus, muito importante. A fim de incrementar o controle biológico na cultura, o objetivo deste trabalho foi selecionar em laboratório os isolados mais virulentos de 32 isolados do fungo entomopatogênico Beauveria bassiana (Bals.) Vuill. e 18 isolados de Metarhizium anisopliae (Metsch.) Sorok.. Inicialmente, avaliou-se a virulência dos isolados imergindo adultos da broca da erva-mate em suspensão fungica contendo 1×108 conídios mL-1, e posteriormente os que obtiveram maior mortalidade confirmada, foram selecionados para segunda etapa, na qual avaliou-se a virulência e crescimento vegetativo, produção de conídios em meio de cultura, arroz e em cadáveres de broca. Destacaram-se três isolados de B. bassiana (UNIOESTE 4, UNIOESTE 52 e UNIOESTE 64), e um de M. anisopliae (IBCB 352), atingindo mortalidade confirmada igual ou superior a 90% na primeira etapa, sendo estes selecionados para etapa posterior, juntamente com isolado padrão, CG 716. O isolado IBCB 352 de M. anisopliae, apesar de ser o único da espécie a ser selecionado para a segunda etapa, provocou

Entomopathogenic fungi isolates to control the borer of yerba mate (Hedypathes betulinus) Kluger (Coleoptera; Cerambycidae) / Isolados de fungos entomopatogênicos visando ao controle da broca da erva-mate (Hedypathes betulinus) Kluger (Coleoptera; Cerambycidae)

Yerba mate is an important crop of southern Brazil, Argentina and Paraguay, with socio-economic and environmental importance. Due the lack of native herbals, yerba mate has become a monoculture, and as a consequence, insects have become pests, and the borer, Hedypathes betulinus, one of the most important. In order to improve biological control in the crop, this work aims to select in laboratory the most virulent, of 32 isolates of the entomopathogenic fungi Beauveria bassiana (Bals.) Vuill. and 18 isolates of Metarhizium anisopliae (Metsch.) Sorok. Initially, isolates virolence was evaluated by immersing the adults borers in a fungal suspension, containing 1×108 conidia mL-1, and after, those caused highest confirmed mortality were selected for the second step, wherein it was evaluated the virulence, vegetative growth and conidia production in culture medium, rice and borer bodies. Three isolates of B. bassiana (UNIOESTE 4, UNIOESTE 52 and UNIOESTE 64) and one of M. anisopliae (IBCB 352) reached confirmed mortality equal or superior to 90%, these isolates were selected for the second step, together with the standard isolated GC 716 (B. bassiana). The isolated IBCB 352 of M. anisopliae, despite being the only of this species to be selected for the second step, promoted the high confirmed mortality and conidiogenesis in adults borer, indicating a high inoculums production i, bei

A erva-mate é uma cultura importante da região sul do Brasil, Argentina e Paraguai, tendo grande importância sócio-econômica e ambiental. Com a escassez de ervais nativos, a cultura da erva-mate passou a ser cultivada em sistema de monocultivo e como consequência, alguns insetos tornaram-se pragas, sendo a broca, Hedypathes betulinus, muito importante. A fim de incrementar o controle biológico na cultura, o objetivo deste trabalho foi selecionar em laboratório os isolados mais virulentos de 32 isolados do fungo entomopatogênico Beauveria bassiana (Bals.) Vuill. e 18 isolados de Metarhizium anisopliae (Metsch.) Sorok.. Inicialmente, avaliou-se a virulência dos isolados imergindo adultos da broca da erva-mate em suspensão fungica contendo 1×108 conídios mL-1, e posteriormente os que obtiveram maior mortalidade confirmada, foram selecionados para segunda etapa, na qual avaliou-se a virulência e crescimento vegetativo, produção de conídios em meio de cultura, arroz e em cadáveres de broca. Destacaram-se três isolados de B. bassiana (UNIOESTE 4, UNIOESTE 52 e UNIOESTE 64), e um de M. anisopliae (IBCB 352), atingindo mortalidade confirmada igual ou superior a 90% na primeira etapa, sendo estes selecionados para etapa posterior, juntamente com isolado padrão, CG 716. O isolado IBCB 352 de M. anisopliae, apesar de ser o único da espécie a ser selecionado para a segunda etapa, provocou