Sperm and oocyte as carriers for bovine viral diarrhoea virus biotypes during in vitro fertilization.

Bovine viral diarrhoea virus (BVDV) is an important viral agent causing the reproductive failure in cattle. The objectives of the study were to assess the role of male and female gametes, as carriers of cytopathic (CP) and non-cytopathic (NCP) BVDV to embryonic cells during in vitro fertilization. In this respect, sperm and oocytes were separately exposed to concentrations of 104.5 or 105.5 TCID50 /mL CP and NCP BVDV, for 2 h before fertilization. After washing, the intact gametes with the infected gametes were inseminated. Seven days post-fertilization, the virus-exposed embryos were examined for presence of the viral genome by RT-PCR. One-way anova with post-hoc Tukey's HSD test and an independent samples t-test were used to compare within and between groups, respectively. The results presented a significant decrease in the blastocyst rates for CP-infected groups than NCP-infected groups (p ≤ .01). Compared to the controls and the infected oocyte groups, the cleavage rates of the infected sperm groups (NCP and CP BVDV) were significantly reduced both in low (104.5 TCID50 /mL) and high (105.5 TCID50 /mL) titres of the virus (p ≤ .01). The proportion of embryos which was developed to blastocyst stages was significantly lower for CP and NCP-infected groups than the control groups (p ≤ .001). According to the molecular results, all samples of the retarded/degenerated embryos (at least one blastocyst within each one) in CP and NCP groups, one sample (at least one blastocyst in that) within a CP-infected group, and six samples (at least one blastocyst in each one of those) of NCP-infected groups contained the viral nucleic acid. Likewise, the results of viral enrichment showed all reactions in which RT-PCR were positive induced CPEs in MDBK monolayers. In conclusion, it is clear that CP and NCP BVDV were able to traverse zona pellucida during fertilization, and they had also negative effects on embryo development.

Effect of bovine viral diarrhea virus biotypes exposure on bovine gametes in early embryonic development in vitro.

Bovine viral diarrhea virus (BVDV) is an important viral agent causing reproductive failure in cattle. The objectives of the current study were to investigate the interaction between two BVDV biotypes, cytopathic (CP) and Non-cytopathic (NCP) and bovine gametes during in vitro fertilization (IVF) processing, the existence of the virus within embryonic cells and early embryonic development rates. Sperm and ova were exposed separately to CP and NCP BVDV at two concentrations of 104.5 and 105.5 tissue culture infectious dose 50.00% (TCID50) mL-1 prior to IVF, respectively. After five days post-IVF, early embryonic development rates of infected groups were assessed. Several embryos of each group, normal and degenerated, were selected for a viral assay using reverse transcription polymerase chain reaction technique. The result showed that the early embryonic development rates were decreased in treatment groups. The rates in the CP groups were lower than the NCP groups. In the CP groups, the proportions were, respectively, 10.00, 6.00 and 11.00, and 6.00% in the infected sperm and oocyte groups (104.5 and 105.5 TCID50 mL-1) that were higher than 50.00% in the control group. In NCP groups, the rates were, respectively, 25.00, 18.00 and 24.00, and 21.00% in the infected groups compared to 48.00% in the control group. In the CP groups, no BVDV was detected in normal embryos, whereas, all degenerated embryos were completely virus-positive. In the NCP groups, the virus was detected in both normal and degenerated embryos. In conclusion, this study supported detrimental impacts of CP and NCP BVDV on early embryonic development and the role of sperm and the zona pellucida layer as carriers of the virus.

Long Term Oral Administration of Oregano Essence Effectively Relieves Polycystic Ovarian Rats through Endocrine and Inflammatory Balance.

Polycystic ovarian syndrome (PCOS) is alarmingly rising and sustainable therapy/prevention is needed. Here, we evaluated the therapeutic effects of oregano or Origanum vulgare (O. vulgare) essence (OE) on the PCOS rat model system. Vaginal smears monitored the estrous cycle of 40 virgin adult rats, and they received 2 mg estradiol valerate (EV)/0.2 ml corn oil intramuscularly to induce PCOS. At 60 days post-EV injection, all rats were evaluated for follicular development/cysts. The EV-induced PCOS rats were orally administered 250 and 500 mg/kgBW/day of OE for 30 days. OE was also further assessed for its predominant components along with hormonal, histological, and inflammatory-related gene expressions in the ovaries. The main components of the OE were predominantly pulegone (36.3), L-menthone (31.3%), far less piperitone (7.8%), isopiperitone (6.4%), isomenthol (3.6%), humulene epoxide II (2.2%), α-pinene (1.7%), and thymol (1.5%). Hormonal, histological, and inflammatory-related gene expression results showed >4-fold and 1.5-fold increase in FSH and progesterone; ∼50%, 85%, 45%, 55%, and 30% decreased in LH, estradiol, estrogen, testosterone, and AMH; and dose-dependently decreased in mRNA expression of IL-6, IL-1α, NF-kB, TNF-α, and IL-1ß by 25-65%, 55-75%, 15-40%, 30-55%, and 35-55%, respectively, and thus decreased the severity of PCOS, boosted endocrine balance, restored functional follicles and corpus luteum, and thus ovulation in PCOS rats. Overall, in the disrupted PCOS rats, OE oral treatment effectively relieved estradiol-induced PCOS rats via: (1) its endocrine balancing on GnRH, FSH, and LH and (2) its anti-inflammatory and antioxidant properties on ovary caused by OE's useful compounds like pulegone, thymol, and L-menthone. Though many aspects of the effects remain to be tested, such an underlying mechanistic reproductive regulatory effect observed in OE-administered rats further proves its sensible pharmaceutical applications in reproductive medicine and more specifically, PCOS.

Seroprevalence of leptospiral infection in feline population in urban and dairy cattle herds in Mashhad, Iran.

The importance of cats in the Leptospira epidemiology is due to the possibility of transferring leptospirosis to wild and domesticated animals. The purpose of this survey was to determine the prevalence of Leptospira infection in shorthair cats in different location of Mashhad, Iran. Totally, 147 blood samples were taken from 42 (28.57%), 52 (35.37%) and 53 (36.05%) households, stray and cats which lived in industrial dairy cattle herds of Mashhad, Iran, respectively. Sera were tested with seven live Leptospira antigens using microscopic agglutination test (MAT). Sera with 50.00% agglutination at the dilution of ≥ 1/100 were considered as positive samples. Agglutination at dilutions of < 1/100 considered as suspected to Leptospira infection. Overall, 19 (12.92%) out of 147 cats showed reaction in MAT. The seroprevalence at a titer ≥ 1:100 and < 1:100 were 10 (6.80%) and 9 (6.12%), respectively. Serum samples showed positive reaction against Leptospira intterogans hardjo (no = 10; 52.63%), pomona (no = 5; 26.31%) and icterohaemorrhagiae (no = 4; 21.05%). Eight cats (42.10%) belong to dairy cattle herds had the most infection only by L. I. hardjo with 1:200 titer. There were no significant differences among the weight' age and sex of infected cats. However, there were significant differences between the infected cats in dairy cattle herds and the cats in the urban area (p < 0.05). It is concluded that cats can be infected by Leptospira spp. especially in commercial dairy cattle herds. Cats can be considered as a sanitation hazards in the area for this zoonotic disease.

Reproductive performance in out-of-breeding season of fatty ewes using implant norgestomet with or without PMSG.

Pregnancy in out-of-breeding season in ewes increases the economical goals. Synchronization of estrus and ovulation is essential for above program. The aims of this study using implant norgestomet with or without Pregnant Mare Serum Gonadotropin (PMSG) were to evaluate the serum progesterone (P4) concentration changes: the conception rate and estimation of the lambing rate and litter size. In total, 80 non-cycling multiparous Iranian Kurdish breed fat-tailed ewes with <0/5 ng/ml P4 were used in April and May 2008 in the suburb of Mashhad, Iran. The animals were randomly divided into three groups: a control group (n = 30) without hormonal treatment, another group (n = 25) received 3 mg of norgestomet implant placed subcutaneously in the convex surface of the ear for 9 days, and the third group (n = 25) treated with 3 mg of norgestomet implant for 9 days with an IM injection of 500 IU PMSG at implant removal. The progesterone of treatment and control groups were measured on days 4, 9, and 13 after removal of the norgestomet using radioimmunoassay. Every five ewes were exposed to one ram after 24 h of norgestomet removal in treatment and control groups, simultaneously. The pregnancy was examined after 25 days of ram removal using ultrasonography. Progesterone concentration was significantly higher in treatment groups on 9 and 13 days after norgestomet removal (P < 0.05). The pregnancy rate in the control, non-PMSG, and PMSG treatments groups were 17%, 52%, and 72%, respectively. The rates of single and twin pregnancy in the non-PMSG treatment group were 69% and 31%, respectively. These rates in norgestomet and PMSG treatment group were 50% and 39%, respectively. Triplet pregnancy (11%) was observed only in the PMSG treatment group. It was concluded that using implant norgestomet especially accompanied with PMSG can increase and improve the fertilization rate of ewe in the out-of-breeding season program.