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Yotov S.A.,Trakia University | Atanasov A.S.,Trakia University | Georgiev G.B.,Trakia University | Dineva J.D.,Institute of Biology and Immunology of Reproduction | Palova N.A.,Experimental Station of Agriculture
Asian Pacific Journal of Reproduction | Year: 2014

Objective: To investigate some blood biochemical parameters in anoestrous dairy cows with cystic ovarian follicles (COF) during the first two months post partum and the effect of hCG-PGF2α -GnRH treatment on their reproductive response. Methods: The investigation was performed on 20 lactating Holstein cows, divided into two groups: control group (n=8) and treated group (n=12). All animals had cystic ovarian follicles, detected during the two consecutive transrectal ultrasound examinations at 7-day interval. Blood samples from each animal were collected on the day of second ultrasonography (Day 0) and subjected to a hormonal and biochemical analysis. Treated cows were subjected to another ultrasound examination on day of prostaglandin injection and the presence of corpus luteum was recorded. The control animals were not treated, while the treated group received 1 500 IU hCG on day 0, PGF2α 500 mg on day 7 and 100 μg GnRH 48 hours latter. Eighteen hours after GnRH administration animals bearing corpus luteum on Day 7 were artificiallly inseminated. The last ultrasound examination was made forty days after start of the threapy. On the base of the obtained results, cumulative oestrous activity, cumulative ovulation rate, pregnancy rate and COF persistence were determined. Results: The blood analysis in the both groups on Day 0 showed significantly (. P<0.05) lower mean concentration of progesterone, glucose and inorganic phosphate compared to the reference range values for cyclic dairy cows. Cumulative oestrous activity (91.7%) and cumulative ovulation rate (75.0%) were considerably higher (. P<0.05) in treated group than control group, whereas the cystic follicle persistence (25.0%) was lower (. P<0.05) in the treated group than control group. Conclusions: The present study confirmed the thesis for low blood levels of glucose and inorganic phosphate in cows presenting cystic ovarian follicles. It also indicated significant improvement of cumulative oestrous activity and ovulation rate in animals subjected to hCG-PGF2α -GnRH administration than non-treated animals. The used hormonal protocol could decrease cystic ovarian follicle persistence in dairy cows. © 2014 Hainan Medical College. Source

Markova N.,Bulgarian Academy of Science | Slavchev G.,Bulgarian Academy of Science | Djerov L.,Medical University-Sofia | Nikolov A.,Medical University-Sofia | Dimova T.,Institute of Biology and Immunology of Reproduction
Human Vaccines and Immunotherapeutics | Year: 2016

Our previous studies showed that mycobacterial L-forms persist in the blood of BCG vaccinated people and that BCG vaccine is able to produce, under appropriate conditions, filterable, self-replicating L-bodies with virus-like size. Because filterability is one of the characteristics of L-forms, considerable interest has been shown in their capacity to cross the maternal-fetal barrier. The current study demonstrated isolation of mycobacterial L-form cultures from umbilical cord blood of 5 healthy newborns of healthy mothers vaccinated previously with BCG. The isolated cultures showed distinctive growth characteristics of cell wall deficient L-form bacteria. Transmission electron microscopy demonstrated presence of L-bodies with extremely small size of 100 nm and revealed morphological transformations, typical for L-forms. IS6110 Real Time PCR assay confirmed that all L-form isolates were of mycobacterial origin and belonged to Mycobacterium tuberculosis complex which includes vaccinal BCG substrains. In conclusion, we could suggest that reproductive filterable L-bodies of BCG origin are able to fall in blood circulation of the fetus by vertical transmitted pathway and colonize newborns. © 2016 Taylor & Francis Source

Rahimi G.,University of Cologne | Isachenko V.,University of Ulm | Kreienberg R.,University of Ulm | Sauer H.,Justus Liebig University | And 4 more authors.
European Journal of Obstetrics Gynecology and Reproductive Biology | Year: 2010

Objective: In ovarian tissue grafts there is a massive loss of follicles during the ischaemic period until re-vascularisation is established. The aim of our study was to investigate the influence of different cryopreservation techniques on the ability for the re-vascularisation of ovarian tissue transplanted to SCID mice. Study design: Ovarian fragments from five patients were cut into pieces (∼0.5 mm × 1.0 mm × 1.0 mm) and randomly distributed into three groups: fresh non-treated tissue (group A); tissue conventionally frozen in standard 0.5 ml insemination straws with 1.5 M ethylene glycol + 0.1 M sucrose, with thawing in a 40 °C water bath and step-wise removal of cryoprotectants at room temperature in 0.5 M, 0.25 M and 0.15 M sucrose with gentle agitation (group B); tissue vitrified in 2.62 M dimethylsulphoxide + 2.6 M acetamide + 1.31 M propylene glycol + 0.0075 M polyethylene glycol, with warming by direct plunging of solid specimens with ovarian pieces into 20 ml of 50% vitrification solution pre-warmed to 40 °C and dilution of cryoprotectants in a decreasing concentration of vitrification solution (25%, 12.5%) at room temperature (group C). We used a xenograft model in which ovarian tissue pieces of all three groups were subcutaneously transplanted in SCID mice. The animals were sacrificed on the third day after ovarian tissue transplantation and then weekly during 1 month to obtain the ovarian tissue grafts. These samples were examined by immunohistochemical staining with the endothelial cell-specific marker platelet endothelial cell adhesion molecule-1 (PECAM-1) to determine angiogenesis. Histological observation of tissue after explantation was performed and quality and quantity of follicles were assessed. Results: No PECAM-1 staining was observed in all treatment groups prior to grafting. After warming and in vivo culture of ovarian tissue, the beginning of angiogenesis in pieces from all treatment groups on the third day was detected by PECAM-1 staining. After 4 weeks of in vivo culture the overall area of PECAM-1-positive blood vessels significantly increased (P < 0.05), independent of the type of cryopreservation (groups B and C vs. group A). It was found that transplantation technique had negative influence on the integrity of follicles independent of the type of treatment during in vivo culture. The duration of in vivo culture has a negative, but not statistically significant, influence on follicle quality in long-cultured transplants inside each treatment group (P > 0.5). Conclusion: The process of re-vascularisation of transplanted ovarian tissue is independent of the type of treatment and does not influence follicle quality. © 2009 Elsevier Ireland Ltd. All rights reserved. Source

Vachkova E.,Trakia University | Bosnakovski D.,University of Macedonia | Yonkova P.,Trakia University | Grigorova N.,Trakia University | And 7 more authors.
In Vitro Cellular and Developmental Biology - Animal | Year: 2016

Rabbits are considered as appropriate animal models to study some obesity-associated abnormalities because of the similarity of their blood lipid profile and metabolism to humans. The current study was focused on comparison of adipose differentiation ability in rabbit adipose-derived stem cells (ADSC) in vitro. Subcutaneous and visceral stromal vascular fractions (SVF) were isolated from three 28-d-old New Zealand rabbits by collagenase digestion. Supernatants from both isolates were collected 24 h after the initial plating. On the fourth passage, all isolated cell types undergo triplicate adipogenic induction. The adipose induction potential was calculated as percentage of increasing optical density (OD) values. The data revealed that with increasing the number of induction cycles, the induction tendency in visceral ADSC decreased in contrast to the subcutaneous ones. Although the supernatants did not reach induction levels of their relevant precursors, they follow the same pattern in both subcutaneous and visceral ADSC. All cell types successfully passed osteogenic and chondrogenic differentiation. In conclusion, the best adipose induction ability was observed in directly plated subcutaneous cell population. The increase of induction numbers depressed adipose induction ability in cell populations derived from visceral fat depots. © 2016 The Society for In Vitro Biology Source

Abadjieva D.,Institute of Biology and Immunology of Reproduction | Grigorova S.,Institute of Animal Science | Petkova M.,Institute of Animal Science
Asian Journal of Animal and Veterinary Advances | Year: 2016

Objective: The objective of the present study was to investigate the effect of jodis concentrate (Ukraine) on testicular morphometry and histological changes in rabbit bucks. Methodology: An experiment was conducted with a total of 30 white New Zealand weaned male rabbits with average body weight of 1.5 kg. The animals were distributed in three groups viz., a control and two experimental groups, with 10 animals in each. The growing rabbits from both experimental groups received jodis concentrate in a dose of 2 mL LG1 of drinking water. Results: The rabbits from the 1st experimental group were born from mothers not supplemented with jodis concentrate during the pregnancy and the sucking periods. The animals from the 2nd experimental group were born from mothers who were given the tested product during the pregnancy and sucking periods. The trial lasted for 48 days. Testes were weighed and prepared for histological examination. Conclusion: The iodine from jodis concentrate in a dose of 2 mL LG1 of drinking water has a potential to improve the testicular morphometry and histology of experimental rabbit bucks born from mothers who were not given the tested additive. On the other hand, a clear negative impact was observed on the morphological and histological changes in the testes of rabbit bucks born from mothers supplemented with jodis concentrate during pregnancy. Therefore, serious attention should be paid to the total level of iodine in the feed for rabbits intended for breeding. © 2016 D. Abadjieva et al. Source

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