High Genomic Variability in Equine Infectious Anemia Virus Obtained from Naturally Infected Horses in Pantanal, Brazil: An Endemic Region Case.

Equine infectious anemia virus (EIAV) is a persistent lentivirus that causes equine infectious anemia (EIA). In Brazil, EIAV is endemic in the Pantanal region, and euthanasia is not mandatory in this area. All of the complete genomic sequences from field viruses are from North America, Asia, and Europe, and only proviral genomic sequences are available. Sequences from Brazilian EIAV are currently available only for gag and LTR regions. Thus, the present study aimed for the first time to sequence the entire EIAV genomic RNA in naturally infected horses from an endemic area in Brazil. RNA in plasma from naturally infected horses was used for next-generation sequencing (NGS), and gaps were filled using Sanger sequencing methodology. Complete viral genomes of EIAV from two horses were obtained and annotated (Access Number: MN560970 and MN560971). Putative genes were analyzed and compared with previously described genes, showing conservation in gag and pol genes and high variations in LTR and env sequences. Amino acid changes were identified in the p26 protein, one of the most common targets used for diagnosis, and p26 molecular modelling showed surface amino acid alterations in some epitopes. Brazilian genome sequences presented 88.6% nucleotide identity with one another and 75.8 to 77.3% with main field strains, such as EIAV Liaoning, Wyoming, Ireland, and Italy isolates. Furthermore, phylogenetic analysis suggested that this Brazilian strain comprises a separate monophyletic group. These results may help to better characterize EIAV and to overcome the challenges of diagnosing and controlling EIA in endemic regions.

Fixed-time insemination with frozen semen in mares: is it suitable for poorly fertile stallions?

The purpose of the present study was to compare two protocols for equine frozen semen programs using either postovulation insemination or fixed-time insemination (FT), evaluating both pregnancy rates and intrauterine fluid (IUF) accumulation after artificial insemination with semen obtained from either highly or poorly fertile stallions. Six ejaculates from two stallions (n = 12) were processed. After thawing, semen samples were evaluated by computerized semen analysis. Fifteen mares (30 cycles) were inseminated with frozen semen from highly fertile stallion A, and 14 mares (28 cycles) were inseminated with frozen semen from poorly fertile stallion B. Ovulations were induced with 1 mg (intramuscular) of deslorelin acetate after the observation of a greater than 35 mm follicle and uterine edema. In postovulation insemination group, mares were inseminated once with 800 × 10(6) total sperm in a maximum 6-hour interval after ovulation. In FT group, mares were inseminated twice with 400 × 10(6) total sperm, 24 and 40 hours after induction. Mares were ultrasonographically examined for IUF accumulation 24 hours and for pregnancy diagnosis 14 days after the last insemination. Although IUF accumulation was more evident in mares inseminated once postovulation, pregnancy rates were similar for both protocols, regardless of the stallion, although a significant effect of the stallion was observed. These results indicated that FTs may be used for both highly and poorly fertile stallions as a practical tool to help spreading the use of frozen semen in equine reproduction programs.

A administração de hCG incrementa a incidência de ovulações duplas e múltiplas em éguas da raça Campolina apresentando folículos codominantes? / Does hCG administration increase the incidence of double and multiple ovulations in Campolina mares with co-dominant follicles?

A gonadotrofina coriônica humana (hCG) é rotineiramente empregada na indução de ovulação em éguas. A incidência de duplas ovulações é variável, sendo comum em determinadas raças brasileiras, como Brasileiro de Hipismo e Campolina. O objetivo deste trabalho foi avaliar se a administração de hCG em éguas da raça Campolina incrementa a incidência de ovulações duplas e múltiplas. Através de palpação retal e ultrassonografia, foram avaliados dois ciclos estrais de 104 éguas, divididas em dois grupos. À detecção de um ou mais folículos ³35 mm e edema endometrial grau 2, 2.500 IU de hCG (IV) eram administrados às éguas do grupo I. O grupo II serviu como controle. Os resultados obtidos foram analisados pelo Qui-quadrado. A incidência de ovulações duplas nos grupos I e II foi de, respectivamente, 32,7% (34/104) versus 30,8% (32/ 104). Ambos os grupos obtiveram incidência semelhante de ovulações triplas (1,9% - 2/104). Apenas uma ovulação quádrupla (1,0% - 1/104) foi detectada, no grupo controle. A incidência de duplas e múltiplas ovulações foi similar entre os grupos (p > 0,05). Pelo fato das éguas da raça Campolina apresentarem, fisiologicamente, considerável incidência de ovulações duplas e múltiplas, é possível que a administração de hCG não tenha influenciado estes padrões ovulatórios.

The human chorionic gonadotrophin (hCG) has been routinely used in the equine species to induce ovulations for more than four decades. The incidence of multiple ovulations in horses is variable, being double ovulations a common occurrence in some Brazilian breeds, such as Brasileiro de Hipismo and Campolina. The aim of this work was to evaluate the role of hCG administration to increase, in Campolina mares, the incidence of double and multiple ovulations, which are desirable in embryo transfer programs. One hundred and four (104) mares had their estrous cycles recorded through rectal palpation and ultrasonographic evaluation during the breeding season of 2003 - 2004, in a total of two hundred and eight (208) cycles (two cycles per mare). Animals were allocated in two groups, with fifty two (52) horses each. In Group I mares, once one or more = 35 mm follicles and grade 2 uterine edema were present, 2.500 IU of hCG were intravenously administered. The untreated mares in Group II served as control. The Chi-Square test was used to analyze obtained results. The incidence of double ovulations in groups I and II was, respectively, of 32.7% (34/104) versus 30.8% (32/104). The incidence of triple ovulations was low in both groups, of 1.9% (2/104). Only once in the control group (1.0% - 1/104) was a quadruple ovulation detected. There was no difference in the incidence of double and multiple ovulations between both groups (p > 0.05). Possibly, hCG administration did not influence ovulation patterns in this work due to the fact that Campolina breed mares physiologically show a considerable incidence of double and multiple ovulations.