Complete Genome Sequence of a Hobi- Like Virus Isolated from a Nelore Cow with Gastroenteric Disease in the State of São Paulo, Brazil Adriana Cortez, a João Pessoa Araújo, Jr., b Eduardo Furtado Flores, c Márcio Garcia Ribeiro, d Jane Megid, d Antonio Carlos Paes, d José Paes de Oliveira Filho, d Leila Sabrina Ullmann, b Camila Dantas Malossi, b Marcos Bryan Heinemann e Universidade Santo Amaro–UNISA, Curso de Medicina Veterinária, São Paulo, Brazil a ; Universidade Estadual Paulista “Júlio de Mesquita Filho”–UNESP, Instituto de Biociências, Botucatu, São Paulo, Brazil b ; Departamento de Medicina Veterinária Preventiva, Universidade Federal de Santa Maria, UFSM, Santa Maria, Rio Grande do Sul, Brazil c ; Faculdade de Medicina Veterinária e Zootecnia, Universidade Estadual Paulista “Júlio de Mesquita Filho”– UNESP, Botucatu, São Paulo, Brazil d ; Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo– USP, São Paulo, Brazil e ABSTRACT The Hobi-like virus presents antigenic and molecular differences in rela- tion to bovine virus diarrhea virus 1 and 2. The description of the complete genome of the Hobi-like virus SV757/15, isolated from a Nelore cow with gastroenteric dis- ease in Brazil, will help in understanding the evolution and diversity of pestiviruses. T he Hobi-like virus BVDV-3, an atypical pestivirus, was first identified in bovine fetal serum from Brazil (1). Although it presents clinical similarity to bovine virus diarrhea virus 1 (BVDV-1) and 2 (BVDV-2) in hosts, it has antigenic and genomic differences that may interfere with diagnosis and protection by vaccination (2–5). The strain SV757/15 was isolated from intestinal fragments from a 32-month-old Nelore cow that presented severe lesions, in Torre de Pedra, São Paulo, Brazil. The virus isolate (infected MDBK cell culture supernatant) was centrifuged at 14,000  g for 10 min at 4°C. It was then filtered with 0.45-m disk filters (Millipore), followed by nuclease treatment (50 U of Ambion Turbo DNase in 150 L with incubation at 37°C for 1 h) and RNA purification using a total RNA purification kit (Norgen Biotek). The RNA concentration was determined using a Qubit fluorimeter (Invitrogen). cDNA was syn- thesized using the RevertAid first-strand cDNA synthesis kit (Thermo Fisher Scientific) and random hexamer primers, followed by double-strand cDNA synthesis using RNase H, T4 DNA polymerase, and T4 DNA ligase (Thermo Scientific), as previously described (6). Nextera XT libraries (Illumina) were prepared using 1 ng of double-stranded cDNA, quantified using a Kapa library quantification kit for Illumina platforms (Kapa Biosys- tems) diluted to 1 nM, and sequenced on the NextSeq system (Illumina) using a NextSeq 500 mid-output kit (150 cycles). The initial quality of each sample was assessed using the “QC Report” tool of CLC Genomics Workbench version 9.1. Based on the initial quality reports, filtration was performed using the “Trim Sequences” tool of CLC Genomics Workbench version 9.1. The filtration parameters consisted of removal of low-quality sequences and sequences containing more than 2 ambiguous nucleotides, as well as fixed trimming of 19 nucleotides (nt) from the 5= end and 5 nt from the 3= end. Reads that were less than 50 nt in length after filtration were discarded. Initial assembly for all of the sequences was performed using the de novo assembly strategy of CLC Genomics Workbench version 9.1. The contigs were used to map to a Hobi-like complete genome sequence Received 20 June 2017 Accepted 28 June 2017 Published 17 August 2017 Citation Cortez A, Araújo JP, Jr, Flores EF, Ribeiro MG, Megid J, Paes AC, de Oliveira Filho JP, Ullmann LS, Malossi CD, Heinemann MB. 2017. Complete genome sequence of a Hobi- like virus isolated from a Nelore cow with gastroenteric disease in the state of São Paulo, Brazil. Genome Announc 5:e00767-17. https:// doi.org/10.1128/genomeA.00767-17. Copyright © 2017 Cortez et al. This is an open- access article distributed under the terms of the Creative Commons Attribution 4.0 International license. Address correspondence to Marcos Bryan Heinemann, marcosbryan@usp.br. VIRUSES crossm Volume 5 Issue 33 e00767-17 genomea.asm.org 1