Melatonin reduces apoptotic cells,SOD2 and HSPB1 and improves the in vitro production and quality of bovine blastocysts

Effects of adding different concentrations of melatonin (10^?7, 10^?9 and 10^?11 M) to maturation (Experiment 1; Control, IVM  + 10^?7, IVM  + 10^?9, IVM  + 10^?11) and culture media (Experiment 2; Control, IVC  + 10^?7, IVC  + 10^?9, IVC  + 10^?11) were evaluated on in vitro bovine embryonic development. The optimal concentration of melatonin (10^?9 M) from Experiments 1–2 was tested in both maturation and/or culture media of Experiment 3 (Control, IVM  + 10^?9, IVC  + 10^?9, IVM /IVC  + 10^?9). In Experiment 1, maturated oocytes from Control and IVM  + 10^?9 treatments showed increased glutathione content, mitochondrial membrane potential and percentage of Grade I blastocysts (40.6% and 43%, respectively). In Experiment 2, an increase in the percentage of Grade I blastocysts was detected in IVC  + 10^?7 (43.5%; 56.7%) and IVC  + 10^?9 (47.4%; 57.4%). Moreover, a lower number and percentage of apoptotic cells in blastocysts were observed in the IVC  + 10^?9 group compared to Control (3.8 ± 0.6; 3.6% versus 6.1 ± 0.6; 5.3%). In Experiment 3, the IVC  + 10^?9 treatment increased percentage of Grade I blastocysts with a lower number of apoptotic cells compared to IVM /IVC  + 10^?9 group (52.6%; 3.0 ± 0.5 versus 46.0%; 5.4 ± 1.0). The IVC  + 10^?9 treatment also had a higher mRNA expression of antioxidant gene (SOD 2) compared to the Control, as well as the heat shock protein (HSPB 1) compared to the IVM  + 10^?9. Reactive oxygen species production was greater in the IVM /IVC  + 10^?9 treatment group. In conclusion, the 10^?9 M concentration of melatonin and the in vitro production phase in which it is used directly affected embryonic development and quality.     

Randomly primed,strand-switching,MinION-based sequencing for the detection and characterization of cultured RNA viruses

RNA viruses rapidly mutate, which can result in increased virulence, increased escape from vaccine protection, and false-negative detection results. Targeted detection methods have a limited ability to detect unknown viruses and often provide insufficient data to detect coinfections or identify antigenic variants. Random, deep sequencing is a method that can more fully detect and characterize RNA viruses and is often coupled with molecular techniques or culture methods for viral enrichment. We tested viral culture coupled with third-generation sequencing for the ability to detect and characterize RNA viruses. Cultures of bovine viral diarrhea virus, canine distemper virus (CDV), epizootic hemorrhagic disease virus, infectious bronchitis virus, 2 influenza A viruses, and porcine respiratory and reproductive syndrome virus were sequenced on the MinION platform using a random, reverse primer in a strand-switching reaction, coupled with PCR-based barcoding. Reads were taxonomically classified and used for reference-based sequence building using a stock personal computer. This method accurately detected and identified complete coding sequence genomes with a minimum of 20× coverage depth for all 7 viruses, including a sample containing 2 viruses. Each lineage-typing region had at least 26× coverage depth for all viruses. Furthermore, analyzing the CDV sample through a pipeline devoid of CDV reference sequences modeled the ability of this protocol to detect unknown viruses. Our results show the ability of this technique to detect and characterize dsRNA, negative- and positive-sense ssRNA, and nonsegmented and segmented RNA viruses.     

Artificial Insemination of Gilts with 1.5 Billion Sperms Stored in Different Periods Associated with Different Pre-ovulatory Intervals

This study evaluated the reproductive performance of gilts inseminated at three intervals before ovulation (0-12, 13-23, 24-30 h) with sperm doses (SD) stored for 0-48 and 96-120 h. A total of 218 PIC Camborough 22 gilts were inseminated once with SD of 1.5 x 10(9) sperms. Pregnant gilts (n = 166) were slaughtered 30.8 +/- 3.7 days after artificial insemination. The number of corpora lutea (CL) and total embryos (TE) was counted. Pregnancy rates (PR) were analysed by chi-square test. TE and embryonic survival (ES), obtained as the ratio between viable embryos and CL, were analysed by GLM procedure (SAS) and mean values were compared by Tukey's test. Pregnancy rate was similar among artificial insemination-ovulation (AIOV) intervals when semen was stored for 0-48 h. However, the lowest PR was observed in the 24-30 h AIOV interval with storage time (ST) of 96-120 h (p < 0.05). There was a significant effect of the interaction between ST and AIOV (p < 0.05) on TE and ES variables. Total embryos and ES did not differ (p > 0.05) among AIOV intervals in ST of 0-48 h. However, gilts inseminated at 24-30 h AIOV interval with ST of 96-120 h showed a reduction of 6.7 embryos (p < 0.05) compared with gilts in the same interval inseminated with semen stored for 0-48 h. ES for the 24-30 h AIOV interval and ST of 96-120 h was lower than that observed in the other groups (p < 0.05).     

Analysis of cerebrospinal fluid from dogs and cats after 24 and 48 hours of storage

OBJECTIVE: To compare differential cell counts and cell characteristics of CSF samples analyzed immediately or after storage for 24 and 48 hours at 4 C with and without the addition of autologous serum. DESIGN: Prospective study. ANIMALS: 36 dogs and 6 cats. PROCEDURE: CSF samples were collected from the cerebellomedullary cistern and divided into 250-microliter aliquots. Slides of CSF samples were prepared by use of cytocentrifugation immediately and after 24 and 48 hours of storage with addition of autologous serum (final concentrations, 11 and 29%). Differential cell counts and number of unrecognizable cells were compared among preparations. RESULTS: Significant differences in the differential cell counts were not detected among samples analyzed before or after storage. Although the number of unrecognizable cells increased with storage time, this did not result in a significant effect on cell distribution or diagnosis. Cells in CSF samples stored with 11% serum more closely resembled cells in fresh samples than did cells in samples stored with 29% serum. CONCLUSIONS AND CLINICAL RELEVANCE: CSF samples collected at veterinary clinics remote from a diagnostic laboratory or during nonoperational hours may be preserved through the addition of autologous serum. Evaluation of such samples is likely to result in an accurate diagnosis for at least 48 hours after collection.     

复方泛菌克对犬细小病毒的临床疗效观察

复方泛菌克是(犬毒必治)一种新型高效复方抗菌、抗病毒药物,能迅速消除动物肠毒素和内毒素,增强胃肠吸收能力,提高机体免疫功能。试验用犬毒必治对30只患犬细小病毒的病犬进行治疗,治愈28只,死亡2只,治愈率为93.3%。