Molecular Characterization, Sequence Analysis, and Taxonomic Position of Newly Isolated Fish Iridoviruses

AbstractWithin the past decade, iridoviruses have been identified as the causative agents of systemic disease in a variety of commercially and recreationally important fish. Here we examine nine iridoviruses from fish, reptiles, and amphibians and demonstrate that all isolates were more similar to frog virus 3, the type species of the genusRanavirus,than to lymphocystis disease virus, the type species of the genusLymphocystivirus.Comparison of viral protein synthesis profiles, restriction endonuclease digestion patterns, and the amino acid sequence of the major capsid protein indicated that iridoviruses isolated from the same geographic region were similar, if not identical, whereas viruses from different areas were distinct. Moreover, using primers complementary to the conserved major capsid protein, we found that both PCR and RT-PCR successfully amplified virus-specific nucleic acid from all nine isolates. These studies demonstrate that the piscine iridoviruses examined here were members of the genusRanavirus,and suggest that surveys of pathogenic “fish viruses” may need to include neighboring amphibian and reptilian populations. In addition, the results indicate that PCR readily identified vertebrate iridoviruses and suggest that PCR will be useful in the diagnosis of fish disease

Phylogenetic relationships in the family Alloherpesviridae

Phylogenetic relationships among herpesviruses (HVs) of mammals, birds, and reptiles have been studied extensively, whereas those among other HVs are relatively unexplored. We have reconstructed the phylogenetic relationships among 13 fish and amphibian HVs using maximum likelihood and Bayesian analyses of amino acid sequences predicted from parts of the DNA polymerase and terminase genes. The relationships among 6 of these viruses were confirmed using the partial DNA polymerase data plus the complete sequences of the terminase, helicase, and triplex protein genes; the position of these viruses among all other sequenced HVs was also investigated using the complete terminase gene. The results established the monophyly of the fish and amphibian HVs (Alloherpesviridae) separate from the HVs of mammals, birds, and reptiles (Herpesviridae) and the single recognized HV of bivalve mollusks (Malacoherpesviridae) in the order Herpesvirales. Two major clades in the family Alloherpesviridae were recognized: one consisting of viruses from cyprinid and anguillid hosts and the other of viruses from ictalurid, salmonid, acipenserid, and ranid hosts. A comparison of virus and host phylogenies suggested that closely related HVs in this family may have coevolved with their hosts, whereas significant codiversification was not apparent for the more distantly related viruses

Ação do cepa e do ácido giberélico na frutificação da videira 'niagara rosada'

Studies were carried out to establish the effects of exogenous growth regulators on Vitis (labrusca x vinifera) 'Niagara Rosada' fruiting. The investigations were done in the Jundiaí Research Station, Agronomic Institute State of São Paulo, always using disease-free vineyards of good productivity. The morphological transformations of clusters were carried out under the following aspects: weight, length and width of cluster; number of berries; weight, length average and width average of berries; length average/width average ratio of berries; number of seeds; length and diameter of secondary rachis. That characteristics were determined at the time of maturity plus the total sugars, total acid, Maturity Index and reducing sugars in samples of all treatments. The experiment were conduced in order to determine the doses that resulted in the most beneficial effects, always using applications by immersion of the inflorescence. The experiment consisted of appplications of (2-chloroethyl) phosphonic acid (CEPA) at concentrations of 50, 100, 250, 500, 1,000 and 2,000 ppm, 14 days before flowering; treatments with gibberellic acid at concentrations of 100 and 200 ppm before full bloom, 10 days after full bloom, and both before plus after full bloom. Treatment with CEPA 100 ppm plus gibberellic acid 100 ppm before full bloom and check treatment were also used. The use of CEPA before flowering at the concentrations used, did not result in good results in 'Niagara Rosada' clusters; applications of gibberellic acid did not differ significantly from the nontreated vines under the conditions studied.Estudou-se o efeito da aplicação, por imersão, do CEPA (ácido 2-cloroetil fosfônico) e do ácido giberélico, 14 dias antes do florescimento, nas características morfológicas da panícula da videira Vitis (labrus-ca x vinifera) "Niagara Rosada". Alguns tratamentos com ácido giberélico foram concluídos com nova aplicação 10 dias após o florescimento. Neste experimento verificou-se que, aplicação do CEPA na concentração de 250 ppm resultou na formação de panículas com a maioria de características indesejáveis. o tratamento misto CEPA 100 ppm + ácido giberélico 100 ppm também promoveu o aparecimento de panículas subdesenvolvidas. Aplicação de ácido giberélico na concentração de 100 ppm em pré e pós-ílorescimento, resultou médias mais elevadas, com relação ao peso da panícula, comprimento da panícula, peso das bagas e comprimento da ráquis. Ácido giberélico na concentração de 100 ppm aplicado em pós-ílorescimento, promoveu uma tendência de aumento nas médias do tratamento quanto ao comprimento médio das bagas, largura média das bagas, largura do engaço e comprimento da ráquila. Devemos considerar porém, que os resultados obtidos não apresentaram diferenças significativas com relação ao controle, quanto às características das frutificações, nas condições de estudo

Effect of temperature on betanodavirus infection in SSN-1 cell line

Viral Encephalopathy and Retinopathy (VER), otherwise known as Viral Nervous Necrosis (VNN), is responsible for frequent outbreaks with high mortality in a wide variety of marine fish species all over the world. The disease mainly affects the larval and juvenile stages which show neurological symptoms due to the typical vacuolating lesions in the brain, spinal cord and retina. The disease is caused by a virus of the Betanodavirus genus; seven viral species have been classified in this genus. This classification is based on the fish species from which the virus was isolated, but recently the isolation of the same virus from different host species has become very frequent. Cross-infection tests highlighted that the capacity of the virus to infect different species is mainly due to the environmental temperature at which the fish are farmed, thus at which the infection occurs, more than the host species itself. Using capsid protein gene analysis it was possible to classify four genotypes of Betanodavirus: Striped Jack Nervous Necrosis Virus (SJNNV), Tigger Puffer Nervous Necrosis Virus (TPNNV), Barfin Flounder Nervous Necrosis Virus (BFNNV) and Redspotted Grouper Nervous Necrosis Virus. Iwamoto et al. (1999) demonstrated the different infection temperature ranges for the four genotypes. Both the infection and disease seem to be very temperature-dependent, in fact most of the outbreaks happen in the summer season. This preliminary study elucidates the in vitro effects of temperature on Betanodavirus infection in the SSN-1 cell line. NNV-infected SSN-1 cells incubated at 10\u201330\ub0C after viral adsorption were observed for cytopathic effect and viral growth in the culture media supernatant and cell pellets were evaluated by titration