Gammacoronavirus and Deltacoronavirus in Quail.

This paper expands on a previous report about coronaviruses in quail. After surveillance carried out in 2009 and 2010, some farmers started vaccinating quail with the Massachusetts avian infectious bronchitis virus serotype. The samples for this study were collected in 2013 from São Paulo state in southeastern Brazil. Pools of trachea, lungs, reproductive tract, kidneys, and enteric contents from quail and laying hens kept in the same farms and from quail-only farms as well as from both healthy birds and those showing infectious bronchitis-like symptoms were sampled in this study. The samples were screened using nested RT-PCR targeting the 3'-untranslated region of the Gammacoronavirus genus. Based on the DNA sequence for the RNA-dependent RNA polymerase (RdRp) gene, the strains isolated from quail clustered within either the Gammacoronavirus or Deltacoronavirus genus, and sequences from both genera were found in one quail sample. The phylogeny based on the partial S1 subunit sequence showed that the gammacoronaviruses detected in quail and layers belonged to the Brazil type. These results suggest that quail are susceptible to Gammacoronavirus and Deltacoronavirus viruses and indicate that the Massachusetts vaccination was not controlling IBV in quail or chickens.

Assessing the economic burden of avian infectious bronchitis on poultry farms in Brazil.

Avian infectious bronchitis (IB), caused by avian infectious bronchitis virus (IBV), is a worldwide endemic disease of chickens that affects all branches of the poultry industry. Multiple geno/serotypes occur, and lowvaccine cross-protection results from the highly divergent IBV types. In view of the lack of consistent data on the economic losses caused by IB and the poor protection resulting from the use of the Massachusetts type as a live vaccine in Brazil, this survey aimed to estimate the losses per 1000 birds in broiler and breeder flocks positive for IBV. Thirty-two different IBV genetic types were found. In breeders, the total loss per 1,000 birds was US $3567.4 and US $4210.8 at 25-26 and 42 weeks old, respectively, whereas in broilers (48 days old), the estimated loss was US $266.3 per 1,000 birds. Taken together, the results show a significant and measurable economic impact on the broiler and breeder industries, with an age-dependent increasing trend and an association with multiple genetic types of the virus.

An Avian coronavirus in quail with respiratory and reproductive signs.

An Avian coronavirus was detected in pools of lungs, tracheas, female reproductive tracts, kidneys, and enteric contents from quail (Coturnix coturnix japonica) and laying hen flocks, with and without infectious bronchitis (IB)-like signs, cohoused in farms located in two states of southeastern Brazil during 2009-2010. Although Avian metapneumovirus subtype B was found in two layers samples, Newcastle disease virus was not found in quail or in hens. Based on DNA sequences for the 3'-untranslated region and the gene encoding the RNA-dependent RNA polymerase, this avian coronaviruses in quail is an IB virus-like gammacoronavirus.

Bovine rotavirus in turkeys with enteritis.

Rotaviruses are the main agents responsible for diarrhea in different animal species and for infantile gastroenteritis. These viruses have been isolated from various avian species and have often been associated with poult enteritis and mortality syndrome. Nevertheless, the knowledge of rotavirus infection in turkeys is scarce. Six group A rotavirus strains obtained from pooled enteric contents of diarrheic turkeys were isolated in MA-104 cell culture and typed as G(6)P(1), a typical bovine rotavirus genotype. Additionally, the electropherotypes showed a migration pattern identical to the Nebraska calf diarrhea virus, and the complete NSP4 gene phylogeny showed that all six strains segregated in the genotype E2. Taken together, these results point toward a cattle-to-turkey rotavirus transmission. As a conclusion, bovine-origin rotavirus can be found in turkeys, and this transmission route must now be considered for the improvement of the health status in turkey farms.

Molecular epidemiology of avian infectious bronchitis in Brazil from 2007 to 2008 in breeders, broilers, and layers.

Multiple lineages of Brazilian strains from 2007 to 2008 of avian infectious bronchitis virus (IBV) were detected in flocks of breeders, broilers, and layers. Organs samples from 20 IBV-positive flocks with variable clinical signs were submitted to the partial amplification of S gene (nucleotides 726-1071) of IBV. Fifteen of the 20 sequenced strains segregated in a unique Brazilian cluster subdivided in three subclusters (Brazil 01, 02, and 03). Whereas three strains could be classified as Massachusetts (Mass) genotype, the remaining two strains, originating from flocks with reproductive and respiratory disorders, grouped within the 4/91-793B genotype, a genotype that has not been detected before in Brazil. The potential relevance of the findings to the poultry industry is discussed because the low level of identity of the sequenced part of the S gene from 17 of 20 detected field strains and the vaccines of the Massachusetts serotype used suggest that the level of cross-protection by the Massachusetts vaccines might be low.

Winter dysentery in cows associated with Bovine Coronavirus (BCoV)

Descreve-se a pesquisa de BCoV e rotavírus em 13 mostras fecais de vacas de surtos de disenteria utilizando uma nested PCR dirigida ao gene RdRp e PAGE, respectivamente. Todas as amostras fecais foram positivas para BCoV e nenhuma delas apresentou-se positiva para rotavírus em PAGE. O encontro de coronavírus bovino em amostras fecais de vacas com disenteria sugere que este vírus possa ser o agente primário envolvido na etiologia dos casos aqui relatados

Orchitis in roosters with reduced fertility associated with avian infectious bronchitis virus and avian metapneumovirus infections.

The pathogenesis of infection involving both infectious bronchitis virus (IBV) and avian metapneumovirus (aMPV) causes reproductive damage in hens after viral replication in the epithelium of the oviduct, resulting in loss of cilia and degeneration and necrosis of the epithelial and glandular cells. Although IBV has been indicated as a possible cause of the formation of calcium stones in the epididymus of roosters, a definitive association has not been confirmed. This report describes the detection of IBV and aMPV in the testes of roosters from a Brazilian poultry broiler breeder's flock with epididymal stones and low fertility. Samples of testis, trachea, and lungs from breeder males aged 57 wk were positive for IBV by reverse transcriptase-polymerase chain reaction (RT-PCR), and virus isolation and testis samples were also positive for aMPV by RT-PCR. The inoculation of testis samples into embryonated chicken eggs via the allantoic cavity resulted in curled, hemorrhagic, and stunted embryos typical of IBV infection. The allantoic fluid was positive by RT-PCR aimed to amplify the region coding for the S1 subunit of the IBV S gene, but it was not positive for aMPV. Sequence analysis of the amplified fragment revealed a close relationship with European IBV genotype D274, previously unreported in Brazil. These results indicate that IBV and perhaps aMPV are likely to have played a role in the pathogenesis of the testicular disease described and should be regarded as factors that can influence male fertility disease in chickens.

Molecular characterization of infectious bronchitis virus strains isolated from the enteric contents of Brazilian laying hens and broilers.

Infectious bronchitis virus (IBV) is the causative agent of avian infectious bronchitis, which is characterized by respiratory, reproductive, and renal signs. However, the role of IBV as an enteric pathogen in still controversial. In Brazil, antigenic groups of IBV divergent from the Massachusetts serotype used for vaccination schedules in that country have already been demonstrated. The present study aimed to assess the different genotypes of IBV in Brazilian commercial poultry flocks by partial sequencing of the S1 amino-terminus coding region using enteric contents as samples and examine their relationship with the vaccine serotype currently in use. Samples of enteric contents were taken as pools of five birds from each of 18 poultry farms (17 broiler and one laying farm) from five Brazilian states between 2002 and 2006. Birds were presenting watery diarrhea and poor general condition but were without respiratory, renal, or reproductive signs. Conventional antibacterial and anticoccidial therapies were unsuccessful and, furthermore, all samples proved negative for rotavirus, reovirus, and astrovirus. Eleven IBV samples were isolated in embryonated eggs and resulted in S1 sequences. Phylogenetic analysis showed that these segregated into an exclusive cluster, close to serotype D274, but distant from Massachusetts. Mean amino acid identity amongst these Brazilian strains was 94.07%; amongst these and serotypes D274, 4/91, and Massachusetts, mean amino acid identity was 77.17%, 69.94%, and 68.93%, respectively. In conclusion, the presence of genotype variant strains of IBV in Brazilian poultry flocks has been demonstrated and might be the reason for the unsuccessful control of IBV in Brazil. Furthermore, these results also strengthen the implications of IBV in enteric diseases of poultry.

Isolamento e identificação de Bacillus spp de leite UHT: detecção de toxina / Isolation and identification of Bacillus Spp of milk UHT: detection of toxin

O leite produzido a ultra alta temperatura, denominado UHT, é o leite de maior aceitação no mercado consumidor brasileiro. Neste estudo, determinou-se o número de microorganismos mesófilos aeróbios estritos e facultativos viáveis em 65 amostras de leite UHT, procedente de indústrias sob Inspeção Federal no Estado de São Paulo, coletadas durante 2002 e primeiro semestre de 2003. Observou-se que sete amostras (10,76 por cento) estavam fora dos padrões. As produções relativas a duas amostras foram inutilizadas pelo controle de qualidade das empresas, enquanto as demais (7,69 por cento) foram para o mercado consumidor, sendo estas provenientes de duas indústrias processadoras. Analisou-se a presença de Bacillus spp, assim como os eventuais esporos sobreviventes ao processamento, foram isoladas oito cepas de Bacillus spp. Utilizou-se também três (3) cepas de Bacillus spp isoladas pelo Instituto Adolfo Lutz, da cidade de São Paulo, de leite UHT alvo de reclamações de consumidores. Após a identificação morfológica, bioquímica e genética de culturas do Bacillus spp, estudou-se a produção de toxinas por estas amostras, através de inoculação de sobrenadante de cultura estéril em alça ileal ligada de coelho, teste de Dean e em células Vero, Hep2 e fibroblasmo de embrião de galinha. A identificação através de espectroscopio infra-vermelha de Fourier (FT-IR) das amostras isoladas pelo Instituto Adolfo Lutz caracterizou Bacillus flexus, enquanto as amostras isoladas de leite procedente de indústrias apresentaram a presença de B. flexus e B. sporothermodurans (...)

Molecular analysis of Brazilian strains of bovine coronavirus (BCoV) reveals a deletion within the hypervariable region of the S1 subunit of the spike glycoprotein also found in human coronavirus OC43.

Bovine coronavirus (BCoV) causes enteric and respiratory dis- orders in calves and dysentery in cows. In this study, 51 stool samples of calves from 10 Brazilian dairy farms were analysed by an RT-PCR that amplifies a 488-bp fragment of the hypervariable region of the spike glycoprotein gene. Maximum parsimony genealogy with a heuristic algorithm using sequences from 15 field strains studied here and 10 sequences from GenBank and bredavirus as an outgroup virus showed the existence of two major clusters (1 and 2) in this viral species, the Brazilian strains segregating in both of them. The mean nucleotide identity between the 15 Brazilian strains was 98.34%, with a mean amino acid similarity of 98%. Strains from cluster 2 showed a deletion of 6 amino acids inside domain II of the spike protein that was also found in human coronavirus strain OC43, supporting the recent proposal of a zoonotic spill- over of BCoV. These results contribute to the molecular characterization of BCoV, to the prediction of the efficiency of immunogens, and to the definition of molecular markers useful for epidemiologic surveys on coronavirus-caused diseases.