High genetic diversity of alphacoronaviruses in bat species (Mammalia: Chiroptera) from the Atlantic Forest in Brazil.

Bat coronaviruses (Bat-CoVs) represent around 35% of all virus genomes described in bats. Brazil has one of the highest mammal species diversity, with 181 species of bats described so far. However, few Bat-CoV surveillance programmes were carried out in the country. Thus, our aim was to jevaluate the Bat-CoV diversity in the Atlantic Forest, the second biome with the highest number of bat species in Brazil. We analysed 456 oral and rectal swabs and 22 tissue samples from Atlantic Forest bats, detecting Alphacoronavirus in 44 swab samples (9.6%) targeting the RdRp gene from seven different bat species, three of which have never been described as Bat-CoV hosts. Phylogenetic analysis of the amino acid (aa) sequences coding the RdRp gene grouped the sequences obtained in our study with Bat-CoV previously detected in identical or congeneric bat species, belonging to four subgenera, with high aa identity (over 90%). The RdRp gene was also detected in three tissue samples from Diphylla ecaudata and Sturnira lilium, and the partial S gene was successfully sequenced in five tissues and swab samples of D. ecaudata. The phylogenetic analysis based on the partial S gene obtained here grouped the sequence of D. ecaudata with CoV from Desmodus rotundus previously detected in Peru and Brazil, belonging to the Amalacovirus subgenus, with aa identity ranging from 73.6% to 88.8%. Our data reinforce the wide distribution of Coronaviruses in bats from Brazil and the novelty of three bats species as Bat-CoV hosts and the co-circulation of four Alphacoronavirus subgenera in Brazil.

Actinobacteria from Termite Mounds Show Antiviral Activity against Bovine Viral Diarrhea Virus, a Surrogate Model for Hepatitis C Virus.

Extracts from termite-associated bacteria were evaluated for in vitro antiviral activity against bovine viral diarrhea virus (BVDV). Two bacterial strains were identified as active, with percentages of inhibition (IP) equal to 98%. Both strains were subjected to functional analysis via the addition of virus and extract at different time points in cell culture; the results showed that they were effective as posttreatments. Moreover, we performed MTT colorimetric assays to identify the CC50, IC50, and SI values of these strains, and strain CDPA27 was considered the most promising. In parallel, the isolates were identified as Streptomyces through 16S rRNA gene sequencing analysis. Specifically, CDPA27 was identified as S. chartreusis. The CDPA27 extract was fractionated on a C18-E SPE cartridge, and the fractions were reevaluated. A 100% methanol fraction was identified to contain the compound(s) responsible for antiviral activity, which had an SI of 262.41. GC-MS analysis showed that this activity was likely associated with the compound(s) that had a peak retention time of 5 min. Taken together, the results of the present study provide new information for antiviral research using natural sources, demonstrate the antiviral potential of Streptomyces chartreusis compounds isolated from termite mounds against BVDV, and lay the foundation for further studies on the treatment of HCV infection.

Fontes de infecção e perfil de suscetibilidade aos antimicrobianos de Salmonella sp. isoladas no fluxo de produção de frangos de corte / Sources of infection and antimicrobial susceptibility profile of Salmonella sp. isolated in broiler production flow

Este trabalho foi desenvolvido com objetivo de pesquisar Salmonella em amostras de fígado, coração, saco da gema e mecônio de pintos de corte de um dia; inglúvios e cecos obtidos em abatedouros e em suabes de arrasto; larvas ou adultos de Alphitobius diaperinus. Complementarmente, determinou-se o perfil de suscetibilidade aos antimicrobianos: amoxicilina (10 mcg), ampicilina (10 mcg), ciprofloxacina (5 mcg), enrofloxacina (5 mcg), florfenicol (30 mcg), neomicina (30 mcg), sulfonamida (300 mcg), tetraciclina (30 mcg) e trimetoprim-sulfametoxazol (25 mcg) dos serovares tipificados isolados. As amostras foram submetidas às análises microbiológicas pelos métodos bacteriológicos convencionais. Salmonella sp. foi isolada em 6,2% (4/64) do fígado, 4,7% (3/64) do coração, 3,1% (2/64) dos sacos da gema e 4,7% (3/64) do mecônio, num total de 4,7% (12/256) (pinto de um dia); em 10,2% (13/128) das amostras ambientais, sendo 9,4% (9/96) de suabes de arrasto 12,5%, (4/32) de larvas e adultos Alphitobius diaperinus e em 4,4% (28/640) das amostras em abatedouros, sendo 6,5% (21/320) dos inglúvios e 2,2% (7/320) dos conteúdos cecais de abatedouro. Salmonella enterica ser. Enteritidis foi identificada em suabes de arrasto e em amostras de Alphitobius diaperinus, enquanto Salmonella enterica ser. Typhimurium foi encontrada nos inglúvios e cecos. Salmonella enterica ser. Enteritidis apresentaram 75% (6/8) de resistência às sulfonamidas e Salmonella enterica ser. Typhimurium 100% (3/3). A amoxicilina foi outro antimicrobiano com elevada frequência de resistência. Adicionalmente, 20,7% (11/53) dos serovares apresentaram resistência simultânea a pelo menos dois princípios ativos. Conclui-se que Salmonella encontra-se amplamente distribuída no fluxo de produção de frangos de corte, e a via vertical continua sendo uma fonte de introdução de Salmonella sp. à cadeia de produção; cama e insetos podem perpetuar e veicular Salmonella de interesse zoonótico no ambiente avícola; a existência de cepas resistentes aos antimicrobianos, bem como a resistência múltipla, constituem ameaça à saúde pública.(AU)

This work aimed at searching for Salmonella in liver, heart, yolk sac and meconium samples of one-day-old chicks, crops and cecum samples from slaughterhouses and drag swabs and Alphitobius diaperinus larvae or adults. It also aimed at determining the susceptibility profile to amoxicillin (10 mcg), ampicillin (10 mcg), ciprofloxacin (5 mcg), enrofloxacin (5 mcg), florfenicol (30 mcg), neomycin (30 mcg), sulfonamide (300 mcg), tetracycline (30 mcg) and trimethoprim-sulfamethoxazole (25 mcg) of typed serovars isolated. The samples were submitted to microbiological analyses by the conventional method. Salmonella sp. was isolated in 4.7% (12/256) of samples of one-day old chicks, in 6.2% (4/64) of the liver, 4.7% (3/64) of the hearts, 3.1% (2/64) of the yolk sacs, 4.7% (3/64) of meconium, and in10.2% (13/128) of the environment samples, being 9.4% (9/96) from drag swabs, 12.5% (4/32) from larvae and adult Alphitobius diaperinus and 4.4% (28/640) of the slaughterhouses samples, being 6.5% (21/320) of crops and 2.2% (7/320) of cecum samples. Salmonella enterica ser. Enteritidis was identified only in drag swabs and Alphitobius diaperinus. Salmonella enterica ser. Typhimurium was observed in the crops and cecum, which have shown high frequency of resistance to sulfonamides, 75% (6/8) and100% (3/30), respectively. Additionally, 20.7% (11/53) of the serovars have shown multiresistance to at least two of the tested drugs. In conclusion, Salmonella can be widely spread in broiler production flow, and the vertical pathway is still a major source of Salmonella insertion in the poultry production chain. The litter and bugs can perpetuate and disseminate Salmonella sp. as well as both the existence of strains resistant to antimicrobial and the occurrence of multiresistance are a threat to public health.(AU)