Genetic analysis of the porcine group B rotavirus NSP2 gene from wild-type Brazilian strains

Group B rotaviruses (RV-B) were first identified in piglet feces, being later associated with diarrhea in humans, cattle, lambs, and rats. In human beings, the virus was only described in China, India, and Bangladesh, especially infecting adults. Only a few studies concerning molecular analysis of the RV-B NSP2 gene have been conducted, and porcine RV-B has not been characterized. In the present study, three porcine wild-type RV-B strains from piglet stool samples collected from Brazilian pig herds were used for analysis. PAGE results were inconclusive for those samples, but specific amplicons of the RV-B NSP2 gene (segment 8) were obtained in a semi-nested PCR assay. The three porcine RV-B strains showed the highest nucleotide identity with the human WH1 strain and the alignments with other published sequences resulted in three groups of strains divided according to host species. The group of human strains showed 92.4 to 99.7% nucleotide identity while the porcine strains of the Brazilian RV-B group showed 90.4 to 91.8% identity to each other. The identity of the Brazilian porcine RV-B strains with outer sequences consisting of group A and C rotaviruses was only 35.3 to 38.8%. A dendrogram was also constructed to group the strains into clusters according to host species: human, rat, and a distinct third cluster consisting exclusively of the Brazilian porcine RV-B strains. This is the first study of the porcine RV-B NSP2 gene that contributes to the partial characterization of this virus and demonstrates the relationship among RV-B strains from different host species

Efeito do dimetildioctadecilamônio na resposta imune humoral e celular de cobaios inoculados com o herpesvírus bovino-1 inativado Effect of dimethyl dioctadecyl ammonium chloride on the humoral and cellular response to inactivated bovine herpesvirus-1 in guinea pigs

Avaliou-se o potencial adjuvante do cloreto de dimetildioctadecilamônio (DDA cloreto) em estimular as respostas imune humoral e celular, do tipo hipersensibilidade cutânea tardia (DTH), em cobaios que receberam preparações de antígeno contendo o herpesvírus bovino tipo 1 (BHV-1) inativado. Os animais foram vacinados com o BHV-1 em cinco diferentes formulações, representadas pelos grupos: A- adjuvante completo / incompleto de Freund, B- hidróxido de alumínio (Al(OH)3), C- DDA cloreto, D- associação Al(OH)3 / DDA cloreto e E- BHV-1 em solução aquosa sem adjuvante. Os animais do grupo F, grupo-controle negativo, foram inoculados apenas com meio de cultivo celular. Os maiores títulos de anticorpos neutralizantes do BHV-1, expressos em log2, foram obtidos nos grupos D, A e C, com títulos de 6,25, 6,0 e 5,25, respectivamente. Na avaliação da DTH, os maiores aumentos na espessura da dobra da pele foram observados nos grupos A (2,4mm), C (1,8mm) e D (1,1mm). O DDA cloreto, utilizado tanto de forma isolada quanto em associação, determinou a potencialização das respostas imunológicas humoral e celular de cobaios imunizados com o BHV-1 inativado.<br>The adjuvanticity of dimethyl dioctadecyl ammonium chloride (DDA chloride) to induce humoral and cell mediated (delayed type hypersensitivity - DTH -) immune responses was assessed in guinea pigs that received antigen preparations containing inactivated bovine herpesvirus type 1 (BHV-1). The animals were vaccinated with five different formulations, containing BHV-1 represented by groups: A- complete/incomplete Freund adjuvant, B- aluminum hydroxide (Al(OH)3), C- DDA chloride, D- association Al(OH)3/DDA and E- BHV-1 in aqueous solution without adjuvant. Group F was the negative control group and the animals received only cell culture medium. The higher titers of BHV-1 neutralizing antibodies, expressed in log2, were obtained in groups D, A and C with values of 6.25, 6.0 and 5.25, respectively. In the DTH assessment, the higher increase in skin thickness was observed in group A (2.4mm), C (1.8mm) and D (1.1mm). These results showed that DDA chloride used alone or in association to other substance boosted the humoral and cell mediated immune responses in guinea pigs immunized with inactivated BHV-1

Diarréia em bezerros da raça Nelore criados extensivamente: estudo clínico e etiológico Diarrhea in Nelore calves: Clinical and etiologic study

A diarréia é considerada uma das principais causas de morbidade e mortalidade de bezerros neonatos. Foram colhidas 100 amostras fecais diarréicas e 30 amostras não diarréicas (grupo controle), de bezerros Nelore com até nove semanas de idade com o objetivo de detectar os enteropatógenos Salmonella spp., Escherichia coli, rotavírus, coronavírus, Cryptosporidium spp. e ovos de helmintos. Enteropatógenos foram detectados em 79,0% das amostras diarréicas e em 70,0% das amostras não-diarréicas. No grupo de bezerros com diarréia, E. coli (69,0%) foi o agente mais freqüentemente isolado, seguido de Cryptosporidium spp. (30,0%), coronavírus (16,0%) e rotavírus (11,0%). No grupo controle, E. coli, Cryptosporidium spp. e coronavírus foram detectados, respectivamente, em 66,7%, 10,0% e 3,3% das amostras. Salmonella spp. e ovos de estrongilídeos não foram encontrados nos dois grupos avaliados. A fímbria K99 foi identificada exclusivamente nas linhagens de E. coli isoladas de bezerros com diarréia (5,8%). Entre os antimicrobianos avaliados "in vitro" a enrofloxacina, a norfloxacina e a gentamicina foram os mais efetivos. O peso dos bezerros aos 210 dias de idade não apresentou diferença significativa entre os animais com e sem diarréia.<br>Diarrhea is considered as one of the main causes of morbidity and mortality in neonates calves. Fecal samples from 100 diarrheic and 30 non-diarrheic (control group) Nelore calves less than 9 weeks old were collected for Salmonella spp., Escherichia coli, rotavirus, coronavirus, Cryptosporidium spp., and for helminth eggs investigation. Enteropathogens were detected in 79.0% diarrheic samples and 70.0% non-diarrheic samples. Among diarrheic calves, Escherichia coli (69.0%) was the most common agent found, following by Cryptosporidium spp. (30.0%), coronavirus (16.0%), and rotavirus (11.0%). In the control group, E. coli, Cryptosporidium spp. and coronavirus were detected in 66.7%, 10.0% and 3.3% of the samples, respectively. Salmonella spp. and strongylids were not found in any of the calves from either group. The K99 fimbrial only was detected in E. coli strains from diarrheic calves (5.8%). Enrofloxacin, norfloxacin, and gentamicin were the most effective among the antimicrobials tested. The weight of 210-day-old calves did not show statistic differences between diarrheic and non-diarrheic calves