Rotavirus A and D circulating in commercial chicken flocks in southeastern Brazil.

Rotavirus (RV) outbreaks can cause significant economic losses in the livestock and poultry industries. Stool samples were collected from asymptomatic laying and broiler chickens from commercial poultry farms in the states of Rio de Janeiro and Espírito Santo in southeastern Brazil for detection of RV species A and D (RVA and RVD, respectively) by reverse transcription polymerase chain reaction. RV was detected in 10.5% (34/325) of samples: 22 (64.7%) were positive for RVA and nine (26.5%) for RVD, while three (8.8%) exhibited coinfections with both viruses. Sequence analysis of a VP6 fragment from seven RVA-positive samples identified the I11 genotype in all samples. Information regarding avian RV epidemiology is still scanty, despite the high prevalence of RV infections in several bird species and subsequent economic impact. Consequently, monitoring infections caused by avian RVs, especially in commercial birds, is essential not only to provide new and relevant information regarding the biology, epidemiology, and evolution of these viruses, but also to facilitate the implementation of preventive measures.

Molecular Characterization of Avian Rotaviruses F and G Detected in Brazilian Poultry Flocks.

Avian rotaviruses (RVs) are important etiologic agents of gastroenteritis in birds. In general, avian RVs are understudied; consequently, there is a paucity of information regarding these viruses. Therefore, the characterization of these viral species is highly relevant because more robust information on genetic, epidemiologic, and evolutionary characteristics can clarify the importance of these infections, and inform efficient prevention and control measures. In this study, we describe partial genome characterizations of two avian RV species, RVF and RVG, detected in asymptomatic poultry flocks in Brazil. Complete or partial sequences of at least one of the genomic segments encoding VP1, VP2, VP4, VP6, VP7, NSP1, NSP4, NSP4, or NSP5 of 23 RVF and 3 RVG strains were obtained, and demonstrated that multiple variants of both RVF and RVG circulate among Brazilian poultry. In this study, new and important information regarding the genomic characteristics of RVF and RVG is described. In addition, the circulation of these viruses in the study region and the genetic variability of the strains detected are demonstrated. Thus, the data generated in this work should help in understanding the genetics and ecology of these viruses. Nonetheless, the availability of a greater number of sequences is necessary to advance the understanding of the evolution and zoonotic potential of these viruses.

Rotavirus F and G circulating in chickens in Southeastern Brazil.

Rotavirus (RV) infections represent a significant cause of enteritis and diarrhea in avian species and pose a major concern for the poultry industry. However, the prevalence of rotavirus infections among birds is poorly understood. Stool samples were collected from laying and broiler hens from commercial poultry farms in the states of Rio de Janeiro and Espírito Santo, Southwest region of Brazil, for detection of rotavirus species F and G (RVF and RVG, respectively) by reverse transcription polymerase chain reaction. RV was detected in 11.7% (38/325) of samples: 35 samples were positive for RVF and 3 for RVG. RVF was detected in 15 samples from Rio de Janeiro and 23 samples from Espírito Santo. RVG was detected in 3 samples from Espírito Santo. All the positive samples were from asymptomatic broiler chickens. The prevalence of RV infection in these flocks was high, especially considering that the birds had no apparent clinical disease. Silent circulation in the herds signifies the need for a continuous surveillance program to guide measures to control and prevent this viral infection. Continuous monitoring of pathogens is crucial to ensure greater productivity on poultry farms.

Human polyomaviruses 10 and 11 in faecal samples from Brazilian children.

The human polyomaviruses (HPyVs) 10 and 11 have been detected in faecal material and are tentatively associated with diarrhoeal disease. However, to date, there are insufficient data to confirm or rule out this association, or even to provide basic information about these viruses, such as how they are distributed in the population, the persistence sites and their pathogenesis. In this study, we analysed stool specimens from Brazilian children with and without acute diarrhoea to investigate the excretion of HPyV10 and HPyV11 as well as their possible associations with diarrhoea. A total of 460 stool specimens were obtained from children with acute diarrhoea of unknown aetiology, and 106 stool specimens were obtained from healthy asymptomatic children under 10 years old. Samples were collected during the periods of 1999-2006, 2010-2012 and 2016-2017, and found previously to be negative for other enteric viruses and bacteria. The specimens were screened for HPyV10 and HPyV11 DNA by the polymerase chain reaction (PCR). Randomly selected positive samples were sequenced to confirm the presence of HPyV10 and HPyV11. The sequenced strains showed a percent of nucleotide identity of 93.4-99.6% and 85.5-98.9% with the reference HPyV10 and HPyV11 strains, respectively, confirming the PCR results. HPyV10 and HPyV11 were detected in 7.2% and 4.7% of the stool specimens from children with and without diarrhoea, respectively. The prevalence of both viruses was the same among children with diarrhoea and healthy children. There was also no difference between boys and girls or the degree of disease (severe, moderate or mild) among groups. Phylogenetic analysis showed that all of the genotypes described so far for HPyV10 and HPyV11 circulate in Rio de Janeiro. Our results do not support an association between HPyV10 and HPyV11 in stool samples and paediatric gastroenteritis. Nevertheless, the excretion of HPyV10 and HPyV11 in faeces indicates that faecal-oral transmission is possible.

Identification, characterization and in silico ADMET prediction of Roflumilast degradation products.

The present study reports the degradation behavior of roflumilast (RFL), a new drug developed for the treatment of chronic obstructive pulmonary disease. The degradation of RFL was tested under various stress conditions as per the guidelines of the International Conference on Harmonization. The degradation products (DPs) of RFL were identified, characterized and in silico predictions were made of their pharmacokinetic properties, absorption, distribution, metabolism, excretion and toxicity (ADMET). RFL was subjected to various stress conditions including photodegradation, alkaline and acidic hydrolysis, oxidative and metallic degradation. After analysis by HPLC-DAD, the DPs were isolated by preparative TLC and characterized by high resolution mass spectrometry (HRMS), 1H NMR, 13C NMR and infrared (IR) spectroscopy. RFL tablets were prepared by the addition of solid stressing substances such as excipients and storage in an accelerated stability chamber (40°C; 75% r.h.) for sixteen months. Resulting DPs from the tablets were analyzed by UFLC-QTOF. The most drastic degradation conditions for RFL were 5M NaOH(aq), 6M HCl(aq), 7.5% v/v peracetic acid, which resulted in the isolation of four DPs. However, milder degradation conditions (1M NaOH(aq) and photolysis) generated six DPs (DP-1, 2, 3, 5, 7 and 8), and are more similar to the actual conditions the drug will be exposed. For tablets containing RFL exposed to an alkaline reagent, two DPs were formed: DP-1 and DP-11. Whereas RFL-containing tablets exposed to acid and oxidizing agents, formed one product DP-11. Forced degradation of RFL led to the formation of eleven DPs, seven of which have never been previously reported. RFL is stable under metallic stress and it is relatively stable during photodegradation testing. The UFLC-QTOF methodology detected a greater number of DPs that formed during the stress conditions tested when compared to the HPLC-DAD methodology. In silico prediction of the ADMET properties of the RFL degradation products and metabolites produced in this study are potentially hepatotoxic.