Catedra de Teriogenologia

Slovenia

Catedra de Teriogenologia

Slovenia
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Konrad J.L.,CONICET | Moore D.P.,CONICET | Crudeli G.,Catedra de Teriogenologia | Caspe S.G.,Instituto Nacional de Tecnologia Agropecuaria | And 8 more authors.
Veterinary Parasitology | Year: 2012

The aim of this study was to characterize the pathogenesis of Neospora caninum in experimentally inoculated pregnant water buffalo (Bubalus bubalis). Twelve Mediterranean female water buffaloes ranging in age from 4 to 14 years old and seronegative to N. caninum by indirect fluorescent antibody test (IFAT) were involved. Ten females were intravenously inoculated with 10 8 tachyzoites of NC-1 strain at 70 (n=3) or 90 (n=7) days of pregnancy (dp). Two control animals were inoculated with placebo at 70 and 90dp, respectively. Serum samples were obtained weekly following inoculation to the end of the experiment. Three animals inoculated at 70dp were slaughtered at 28days post inoculation (dpi), three animals inoculated at 90dp were slaughtered at 28dpi and the remaining four animals inoculated at 90dp were slaughtered at 42dpi. Fetal fluids from cavities and tissue samples were recovered for IFAT and histopathology, immunohistochemistry and PCR, respectively. Genomic DNA from fetal tissues was used for parasite DNA detection and microsatellite genotyping in order to confirm the NC-1 specific-infection. Dams developed specific antibodies one week after the inoculation and serological titers did not decrease significantly to the end of the experiment. No abortions were recorded during the experimental time; however, one fetus from a dam inoculated at 70dp was not viable at necropsy. Specific antibodies were detected in only two fetuses from dams inoculated at 90dp that were slaughtered at 42dpi. No macroscopic changes in the placentas and organs of viable fetuses were observed. Nonsuppurative placentitis was a common microscopic observation in Neospora-inoculated specimens. Microscopic fetal lesions included nonsuppurative peribronchiolar interstitial pneumonia, epicarditis and myocarditis, interstitial nephritis, myositis and periportal hepatitis. Positive IHC results were obtained in two fetuses from dams inoculated at 70dp and slaughtered at 28dpi. N. caninum DNA was detected in placentas and fetuses from all inoculated animals. The pattern of amplified microsatellites from placental and fetal tissues resembled the NC-1 strain. Water buffaloes, like cattle, are susceptible to experimental inoculation with N. caninum at early pregnancy. © 2011 Elsevier B.V.


Trasorras V.,Catedra de Teriogenologia | Giuliano S.,Catedra de Teriogenologia | Giuliano S.,Institute Investigacion y Tecnologia en Reproduccion Animal INITRA | Chaves G.,Catedra de Teriogenologia | And 9 more authors.
Reproduction in Domestic Animals | Year: 2012

Contents: The aim of this study was to carry out in vitro fertilization using spermatozoa selected with Androcoll-E™ and to evaluate the efficiency of the culture medium DMEM-F12 for in vitro embryo development in the llama. Twelve adult females from 18 superstimulated (67%) were used as oocyte donors. They were superstimulated with 1500IU of eCG and after 5days, received a single dose of buserelin. Twenty hours post-injection, follicular aspiration was conducted by flank laparotomy. Semen collections were performed under general anesthesia by electroejaculation of the male. The ejaculates were processed with a solution of collagenase (0.1%) and an Androcoll-E™ column was used to improve the sample. Sixty nine COCs were recovered from 79 aspirated follicles (87% recovery). Only expanded COCs were used (n=67); they were randomly placed in groups of 1-5 in Fertil-TALP and the sperm suspension (20×10 6 live spermatozoa/ml) was added to each fertilization microdroplet. After 24h, they were randomly placed in one of two culture media: SOF (n=34) or DMEM-F12 (n=33) and incubated for 6days in humidified atmosphere of 5% CO 2, 5% O 2 and 90% N 2 at 38°C. The blastocyst rate was 20% (7/34) in SOF medium (3 hatched, 2 expanded and 2 early blastocysts) and 15% (5/33) in DMEM medium (all expanded blastocysts). In conclusion, using Androcoll-E™ it is possible to select good quality spermatozoa from llama ejaculates for in vitro fertilization and to produce blastocysts in DMEM-F12 medium. This is also the first time that hatched llama blastocysts have been produced after culture in a defined medium such as SOFaa. © 2011 Blackwell Verlag GmbH.

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