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Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico?CNPq (141398/2015-9). University of Sao Paulo. School of Veterinary Medicine and Animal Science. Department of Pathology. Sao Paulo, SP, Brazil / Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Federal Rural University of the Amazon. Institute of Animal Health and Production. Bel?m, PA, Brazil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Centro Nacional de Primatas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Centro Nacional de Primatas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Centro Nacional de Primatas. Ananindeua, PA, Brasil. University Sao Paulo. Faculty of Medicine. Department of Pathology. Sao Paulo, SP, Brazil. University of Sao Paulo. School of Veterinary Medicine and Animal Science. Department of Surgery. Sao Paulo, SP, Brazil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. University of Sao Paulo. School of Veterinary Medicine and Animal Science. Department of Pathology. Sao Paulo, SP, Brazil. Background The "starry sky" (SK) liver is ultrasonographic pattern characterized by multiple hyperechogenic foci in hepatic parenchyma. The study aimed to characterize the microscopic hepatic lesions in captive owl monkeys with SK liver. Methods Thirty-seven clinically healthy owl monkeys had their liver scanned and 18 of them had liver biopsy. Animals with SK and peliosis hepatis (PH) were subjected to immunohistochemical and molecular screening for Bartonella spp. Results SK liver occurred in 59.4% (22/37) of the owl monkeys. Biopsied animals showed steatosis, hydropic degeneration, hemosiderosis, PH, and multifocal granulomatous hepatitis. Two monkeys had SK, granulomatous hepatitis, and PH which were negative for Bartonella spp. Conclusions PH and granulomatous hepatitis associated with hepatocellular degenerative lesions may present as hyperechoic nodular liver lesions consisted of SK liver; therefore, concomitant occurrence of two lesions or more contributed to the hepatic SK pattern among owl monkeys and such cases might be clinically monitored.

Hepatitis B virus (HBV) is the prototype of the Orthohepadnavirus genus and represents an important cause of chronic hepatitis, liver cirrhosis, and hepatic cancer in humans worldwide. To verify the occurrence and genetic variability of orthohepadnavirus among neotropical bats, we tested 81 liver samples of New World bats from Sao Paulo State, Southeastern Brazil, collected during 2012. PCR, sequencing, and phylogenetic analysis of Surface/Polymerase and Core viral genes confirmed the occurrence of the first isolate of bat orthohepadnavirus detected in South America. These results may contribute to subsequent studies of the origin, variability, host species, and evolution of bat orthohepadnaviruses in South America.

Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico (CNPq) Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil / University of S?o Paulo. Faculty of Veterinary Medicine and Animal Science. Department of Pathology. S?o Paulo, SP, Brazil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. University of S?o Paulo. Faculty of Veterinary Medicine and Animal Science. Department of Pathology. S?o Paulo, SP, Brazil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. We describe a case of microscopic liver lesion caused by larval cestode in a frugivorous bat (Molossus molossus) from the Western Brazilian Amazon. Histopathological analysis of liver indicated the occurrence of metacestode associated with multifocal histiocytic response and the phylogenetic analysis of Cox-I and 18S rDNA genes indicated that the parasite belonged to the family Paruterinidae. This is the first identification of cestodes of the family Paruterinidae in bats and may suggest a broad range of paruterinid tapeworm hosts to be investigated in the Brazilian Amazon.

To date, nothing is known about the genetic diversity of the Echinococcus neotropical species, Echinococcus vogeli and Echinococcus oligarthrus. Here we used mitochondrial and nuclear DNA sequence polymorphisms to uncover the genetic structure, transmission and history of E. vogeli in the Brazilian Amazon, based on a sample of 38 isolates obtained from human and wild animal hosts. We confirm that the parasite is partially synanthropic and show that its populations are diverse. Furthermore, significant geographical structuring is found, with western and eastern populations being genetically divergent.

Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Bel?m, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Bel?m, PA, Brasil/ Universidade de S?o Paulo. Faculdade de Medicina Veterin?ria e Zootecnia. Departamento de Patologia. S?o Paulo, SP, Brazil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Bel?m, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Bel?m, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Bel?m, PA, Brasil/ Universidade de S?o Paulo. Faculdade de Medicina Veterin?ria e Zootecnia. Departamento de Patologia. S?o Paulo, SP, Brazil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Bel?m, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Bel?m, PA, Brasil. Universidade de S?o Paulo. Faculdade de Medicina. Instituto de Medicina Tropical. Laborat?rio de Gastroenterologia e Hepatologia Tropical. S?o Paulo, SP, Brazil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Bel?m, PA, Brasil. The bush dog (Speothos venaticus) acts as the natural definitive host in the life cycle of Echinococcus vogeli, the causative agent of polycystic hydatid disease, a zoonotic neglected disease in the South America. We report a case of natural infection by Echinococcus vogeli in a bush dog from the Brazilian Amazon, confirmed by the morphological and morphometric examination of adult parasites and their hooks obtained from the small intestine of the canid. Additionally, mitochondrial DNA sequence analysis corroborated these findings. This is the second report of natural infection by E. vogeli in a bush dog.

The human Pegivirus (HPgV, also known as GBV-C virus or hepatitis G virus) is a lymphotropic RNA-virus phylogenetically related to the Hepatitis C virus, which infects approximately 5% of the world's human population. Recently, two novel, presumably hepatotropic, pegiviruses, designated as equine Pegivirus (EPgV) and Theiler's Disease Associated Virus (TDAV), were discovered in horses with clinical and laboratory evidence of hepatic disease. To verify the occurrence of pegiviruses infection in horses from Para State, northern Brazil, serum samples from 114 horses located in four cities (Acara, Belem, Dom Eliseu and Ananindeua) were submitted for the molecular analysis of EPgV by nested RT-PCR. The results of nucleotide sequencing and phylogenetic analysis of EPgV NS3 and NS5B genomic regions confirmed one positive sample among 114 tested samples (1/114; 0.8%). No evidence of TDAV infection was found, but despite the low prevalence and unknown clinical significance among the studied population, these results represent the first molecular detection of EPgV in horses in South America.

To the Editor: An estimated ≈150 million persons (3% of the world population) are chronically infected with hepatitis C virus (HCV). This virus is the prototype of the genus Hepacivirus and a major cause of liver cirrhosis and hepatocellular carcinoma around the world. Every year, 3–4 million persons become infected with HCV, and ≈350,000 die of this infection (1). Development of an HCV vaccine has been hampered by the difficulty of in vitro cultivation of the agent and by lack of animal models for studies of viral functions and host immune reactions (1,2). The only effective experimental model that can be infected with HCV and in which the course of infection is similar to that in humans is the chimpanzee. Since the discovery of HCV-like virus in dogs with respiratory disease and nonspecific gastrointestinal disorder in the United States in 2011, tentatively named canine hepacivirus (2), new hepaciviruses have been detected in insectivorous bats (3), Old World monkeys (4), wild rodents (5,6), and domestic horses (6–8). These animals could potentially serve as HCV models, but the accuracy of HCV tissue tropism, pathology, and immunology in natural hosts needs to be demonstrated. No official nomenclature for these recently described hepaciviruses has been defined by the International Committee of Viral Taxonomy. In this article, we refer to the viruses detected in horses as conventional nonprimate hepaciviruses (NPHVs). The aim of this study was to verify NPHV infection in horses from 8 locations in the eastern Brazilian Amazon. During January 2011–November 2013, serum samples were collected from 300 equids in 7 cities and 1 district of the State of Para, Brazil (Technical Appendix). Samples came from 265 horses (Equus caballus), 30 mules (Equus mulus), and 5 donkeys (Equus asinus). All procedures for obtaining and using the samples were approved by the Ethics Committee on the Use of Animals in Research from the Evandro Chagas Institute (protocol code 0023/2012 CEUA/IEC/CENP/SVS/MS). Viral RNA was extracted from serum samples by using the TRIzol LS Reagent (Invitrogen, Carlsbad, CA, USA), and a target sequence in the nonstructural 3 protein (NS3) region of the NPHV genome was amplified from the cDNA by using a previously reported nested PCR protocol (8). Second-round product reactions of ≈380 bp were considered positive. We used the BigDye Terminator version 3.1 Cycle Sequencing Kit and an ABI 3500 Genetic Analyzer (both from Applied Biosystems, Foster City, CA, USA) for sequencing. To obtain consensus sequences, we used BioEdit software, version 7.0.5.3 (http://www.mbio.ncsu.edu/BioEdit/BioEdit.html). To phylogenetically analyze the virus sequences based on an NS3 partial nucleotide sequence of 294 bp, we used the maximum-likelihood method with the Kimura 2-parameter model in MEGA version 5.1 (http://www.megasoftware.net) (bootstrap, 1,000 replicates). The nucleotide distance was calculated by using PAUP version 4.0 software (http://paup.csit.fsu.edu/). The sequences obtained in this study have been assigned GenBank accession nos. {"type":"entrez-nucleotide-range","attrs":{"text":"KJ469442-KJ469466","start_term":"KJ469442","end_term":"KJ469466","start_term_id":"619834736","end_term_id":"619834784"}}KJ469442-KJ469466. Of the 300 serum samples, 25 (8.3%) were positive for NPHV by nested reverse transcription PCR; these results were confirmed by nucleotide sequencing. This prevalence of NPHV infection among horses in Brazil is higher than that reported for other countries (6–8). The prevalence among competition horses (19.1%) was remarkably higher than that among farm (6.3%) and cart horses (6.2%). Many possibilities for the higher prevalence in the first group, such as exposure to nonsterile needles shared by a large number of animals, must be considered; however, further studies are needed to investigate this hypothesis. No mules or donkeys tested in this study were infected by NPHV. A similar finding for another study has been reported (8), which might suggest that these species are not natural hosts for NPHV. Until now, it has been impossible to associate Hepacivirus infection in horses with clinical hepatic disease (7,8). All infected horses for which information was available were healthy at the time of sampling. Our finding of high NPHV prevalence among horses is consistent with findings of previous studies in which NPHV-mediated disease was not found, suggesting that the course of infection is chronic and largely asymptomatic, similar to that of HCV infection in humans. The sequences reported in this study shared close genetic relationships with previously reported NPHV sequences from dogs and horses (Figure) (2,6–8). The nucleotide distances of the sequences ranged from 0 to 19.9%. Figure Maximum-likelihood phylogenetic tree of the partial nucleotide sequences of the nonstructural protein 3 region (294 bp) of Hepacivirus. The retrieved sequences from GenBank are indicated by the accession number followed by the species from which each ... Many of these new isolates were grouped according to geographic origin, except for 2 sequences that grouped distantly from other isolates from Neotropical regions. One isolate ({"type":"entrez-nucleotide","attrs":{"text":"KJ469449","term_id":"619834750","term_text":"KJ469449"}}KJ469449), from a Quarter horse, was closely related to NPHV detected in horses in the United States (7); similarity was ≈94%. The other sequence from a Neotropical region ({"type":"entrez-nucleotide","attrs":{"text":"KJ469466","term_id":"619834784","term_text":"KJ469466"}}KJ469466), from a Marajoara horse, grouped closer to the NPHV from horses in Germany (6); similarity was as high as 95%. NPHV, one of the viruses most closely genetically related to HCV, is present in the Neotropics and was identified in the eastern Brazilian Amazon. The infection seems to be enzootic in the studied horse population and to be geographically dispersed in northeastern Para State. Technical Appendix: Locations in the State of Para (eastern Brazilian Amazon), Brazil, where blood samples were collected from 300 equids for nonprimate hepacivirus testing. Click here to view.(178K, pdf)