191 IN VITRO DEVELOPMENT OF BLASTOCYSTS CONTINUES AFTER ARTIFICIAL INFECTION WITH BOVINE HERPESVIRUS TYPE 5
Notice bibliographique
Résumé
Bovine herpesvirus-5 (BoHV-5), which is the second most important infectious brain disease affecting livestock in Latin America, has been detected in bull semen and aborted fetus; however, no reports are found regarding its presence in bovine embryos. Because it has 90% genomic similarity to BoHV-1, it is possible that BHV-5 can infect the genital system after viral reactivation, leading to reproductive disorders. This study was designed to investigate the effects of infection of bovine blastocysts (B) by BoHV-5. Hormones and fetal calf serum were tested by PCR and considered free of virus. Selected oocytes, obtained from ovaries at a local slaughterhouse, were washed in PBS with 10% fetal calf serum (Nutricell®, Campinas, Brazil). The oocytes were transferred to 100-µL drops of maturation medium consisting of TCM 199 (Gibco®, Grand Island, NY, USA), 0.5 µg mL-1 of FSH (Pluset®, Calier, Spain), and 50 µg mL-1 of LH (Lutropin®-V, Bioniche Inc., Belleville, Ontario, Canada) for 24 h at 39°C and 5% CO2 in air. Afterward, frozen semen (500 µL) was thawed and placed on a Percoll gradient (45 and 90%) and centrifuged at 700 × g for 30 min. The resultant pellet was centrifuged (200 × g for 5 min) in TALP medium and the sperm pellet was then diluted in TALP plus phenylalanine (PHE) and heparin. After an 18-h fertilization period, presumptive zygotes were transferred to culture in CR2 medium up to Day 7 post-fertilization. The procedures were carried out under the same conditions previously reported for IVM. A total of 150 intact B were assigned into 2 experimental groups: I (free of virus) and II (with virus; 102 TCID50/mL for a 1-h period). Then, B of both groups were washed and re-cultured for 72 h in drops of CR2 without virus. The percentage of embryos reaching the hatched blastocyst (HB) stage was observed and analyzed by the chi-square test. At this time, HB were fixed to investigate the presence of virus, degree of apoptosis, and oxidative stress. The virus detection was performed by using an in situ hybridization assay with a specific probe to the glycoprotein C gene of BoHV-5 labeled to biotin. The apoptosis was determined by the annexin V, 4', 6-diamidino-2-phenylindole dihydrochloride (DAPI), and propidium iodide (PI) markers, using immunofluorescence technique. The oxidative stress was realized by using monoclonal anti-AOP1 (antioxidant-like protein 1; Sigma®, St. Louis, MO, USA) through immunoassay. More HB (P > 0.05) were found in group II (75.0%) than for group I (55.0%). In both groups, positive signs for the presence of the apoptosis and oxidative stress markers were observed. The mechanism of apoptosis was initiated independently of virus presence, as evidenced by positive signs also observed in group I. However, oxidative stress was intense in group II, suggesting an evident viral effect on the host cell without compromising embryonic development. These findings might indicate that BoHV-5 uses some mechanisms that keep the cell viable to allow its replication, as seen by the greater hatching rate of infected embryos (75%) compared with the control (55%). FAPESP 07/57774-7 and BRASFRIGO (Birigui-SP).
Récupéré en direct depuis OpenAlex et désinversé. Les résumés ne sont pas conservés dans cette base de données : les index inversés représentent 8,6 Go des 9,3 Go de texte de la base, et le serveur dispose de 13 Go libres.
Comment cette classification a été obtenuedéplier
Prédiction machine sur la base complète
Imitation des enseignantsNi prévalence calibrée, ni vérité terrain. Validation humaine à venir. Le volet Gemma est une étiquette directe du modèle pour chaque travail de la base, lue sur la notice réduite au titre. Le volet Codex est un classifieur appris des 10 348 étiquettes directes de Codex et calibré sur les taux pondérés de l'échantillon; les champs sans appui suffisant ne portent aucun appel Codex. Le mode candidate est l'union des deux volets; le consensus est leur intersection. Ces sorties portent le statut machine_predicted_unvalidated et ne sont pas des étiquettes humaines.
Scores du classifieur distillé par catégorie (deux têtes)
| Catégorie | Codex | Gemma |
|---|---|---|
| Métarecherche | 0,001 | 0,000 |
| Méta-épidémiologie (sens strict) | 0,000 | 0,000 |
| Méta-épidémiologie (sens large) | 0,000 | 0,000 |
| Bibliométrie | 0,000 | 0,000 |
| Études des sciences et des technologies | 0,000 | 0,000 |
| Communication savante | 0,000 | 0,000 |
| Science ouverte | 0,000 | 0,000 |
| Intégrité de la recherche | 0,000 | 0,000 |
| Charge utile insuffisante (le modèle a refusé de juger) | 0,002 | 0,001 |
Scores machine (provisoires)
Les deux têtes enseignantes du modèle étudiant, lues sur ce travail. Un score ordonne la base pour la relecture; il n'affirme jamais une catégorie, et le statut de validation accompagne chaque rangée tel quel.
Scores de référence d'un modèle non mature (critères de maturité non atteints, 7 itérations). Un score ordonne; il n'affirme jamais une catégorie.
score_only:v0-immature-baseline · tel quel depuis la passe de notation : score_only signifie que le nombre peut ordonner les travaux, et qu'aucune étiquette de catégorie n'en découleClassification
machine, non validéePrédiction automatique; un appel candidat d’une seule source (Gemma direct ou Codex distillé), pas un consensus.
Le détail, modèle par modèle et score par score, se trouve en fin de page sous « Comment cette classification a été obtenue ».