9 ENDOSCOPY-MEDIATED INTRATUBAL INSEMINATION IN THE COW – A PRELIMINARY REPORT ABOUT THE APPLICATION OF A NOVEL MINIMALLY INVASIVE INSEMINATION TECHNIQUE

2016 ◽  
Vol 28 (2) ◽  
pp. 134
Author(s):  
K. Stein ◽  
V. Havlicek ◽  
S. Papp ◽  
F. Palm ◽  
G. Brem ◽  
...  
Keyword(s):  
Reproductive Tract ◽  
Capillary Tube ◽  
Vaginal Wall ◽  
Uterine Horn ◽  
Female Reproductive Tract ◽  
Glass Capillary ◽  
Cell Stage ◽  
Frozen Semen ◽  
Hoechst Dye ◽  
Fresh Semen

On their long path through the female reproductive tract to the fertilization site, spermatozoa are exposed to diverse influences and hazards of the cervical, uterine, and oviducal environment that naturally select viable sperms for the following fertilization. Consequently, this results in a reduction from several billions of sperms in the ejaculate to a functional sperm reservoir within the range of 102 in the isthmus of the Fallopian tube. A technique to deposit spermatozoa directly into the ampulla, thus bypassing most of the reproductive tract, enables a rigorous reduction in number of sperms deposited. Furthermore, it provides a direct assessment of sperm fertility. The aim of our study was to establish an endoscopy-assisted intratubal insemination technique using different sperm dosages, fresh or cryopreserved, to determine adequate conditions for optimal fertilization. Eighteen Simmental heifers were inseminated with fresh semen, and 9 heifers were inseminated with frozen semen using this novel technique. The heifers were synchronized using a modified Ovsynch protocol, and insemination was conducted 18 to 20 h after the second gonadotropin-releasing hormone application. Insemination of heifers was performed under epidural anaesthesia. A tubing system bearing the endoscope and an insemination device was introduced through the vaginal wall into the peritoneal cavity. The insemination device consisted of a tube connected to a curved glass capillary tube loaded with semen. After a visual examination of the ovaries for the presence of an ovulatory Graafian follicle, the capillary tube was inserted directly via the infundibulum into the ipsilateral ampulla and the semen dose was deposited. The entire procedure took ~10 min. Two days later the oviduct was flushed by the same technique. A tubing system connected to a metal catheter served for flushing the embryos and unfertilized oocytes from the oviduct into the uterine horn. Afterward, embryos and oocytes were collected by flushing the uterine horn using an embryo flushing catheter and an embryo filter (EmCon). Embryos were stained using a Hoechst dye to visualise the numbers of attached spermatozoa to the zonae pellucidae. From 18 inseminations with fresh semen doses of 7 to 28 million sperms, 7 embryos at the 2- to 8-cell stage were found. Two of these embryos had more than 10 accessory sperms (AS), 3 had 3 to 6 AS, and 2 were without AS. From 9 inseminations with frozen semen doses containing 1.5 million sperms, we obtained 2 embryos, one at the 4-cell stage without AS and one at the 8-cell stage with 5 AS. Additionally, 3 unfertilized oocytes were collected. In conclusion, these preliminary results demonstrate a promising technique for intratubal AI, which has to be further optimized by studying numbers and treatment of spermatozoa and time of insemination.

Surface ultrastructure of danazol treated rat endometrium: A SEM study

1983 ◽  
Vol 41 ◽  
pp. 556-557
Author(s):  
R.P. Apkarian ◽  
J.S. Sanfilippo
Keyword(s):  
Cross Sections ◽  
Reproductive Tract ◽  
Preliminary Investigation ◽  
Breast Disease ◽  
Distal Segment ◽  
Uterine Horn ◽  
Female Reproductive Tract ◽  
Ultrastructural Changes ◽  
Cycle Phase ◽  
Sprague Dawley

The synthetic androgen danazol, is an isoxazol derivative of ethisterone. It is utilized in the treatment of endometriosis, fibrocystic breast disease, and has a potential use as a contraceptive. A study was designed to evaluate the ultrastructural changes associated with danazol therapy in a rat model. The preliminary investigation of the distal segment of the rat uterine horn was undertaken as part of a larger study intended to elucidate the effects of danazol on the female reproductive tract.Cross-sections (2-3 mm in length) of the distal segment of the uterine horn from sixteen Sprague-Dawley rats were prepared for SEM. Ten rats in estrus served as controls and six danazol treated rats were noted to have alterations of the estrus cycle i.e. a lag in cycle phase or noncycling patterns. Specimens were fixed in 3% glutaraldehyde in 0.05M phosphate buffer containing CaCl2 at pH 7.0-7.4 and chilled to 4°C. After a brief wash in distilled water, specimens were passed through a graded series of ethanol, critical point dryed in CO2 from absolute ethanol, and coated with 6nm Au. Observations were made with an IS1-40 SEM operated at 15kV.

scholarly journals Chlamydia muridarum infection differentially changes smooth muscle contractility and responses to prostaglandins in uterus and cervix

2018 ◽  
Author(s):  
Jia Ming Lee ◽  
Jemma R. Mayall ◽  
Anne Chevalier ◽  
Dirk Van Helden ◽  
Jay C. Horvat ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Mrna Expression ◽  
Reproductive Tract ◽  
Uterine Horn ◽  
Female Reproductive Tract ◽  
Muscle Contractility ◽  
Smooth Muscle Contractility ◽  
Contraction Amplitude ◽  
Chlamydia Muridarum ◽  
Reproductive Tract Infection

AbstractChlamydia trachomatis infection is a primary cause of reproductive tract diseases including chronic pelvic pain and infertility. Previous studies showed that this infection alters physiological activities in mouse oviducts. Whether this occurs in the uterus and cervix has never been investigated. This study characterized the physiological activity of the uterus and the cervix in a Chlamydia muridarum (Cmu) mouse model of reproductive tract infection. Uterine or cervix smooth muscle contractility, responses to oxytocin or prostaglandins (PGF2α and PGE2) and mRNA expression of oxytocin and PG receptors were assessed 14 days post infection. Cmu infection did not affect the contractions of the uterine horn but significantly decreased the contraction amplitude of the cervix. Cmu infection did not alter the responses of uterine horn or cervix to oxytocin, however PGF2α induced contractions of the uterine horn, but not the cervix, were significantly increased following Cmu infection. PGE2 contraction amplitude in both the uterine horn and cervix was unaffected by Cmu infection. An upregulation of Ptgfr and a down-regulation of Ptegr4 mRNA expression was observed in the uterine horn following Cmu infection. These results indicate that Cmu infection alters contractility and prostaglandin signalling in the female reproductive tract but the effects are localised to specific regions.

Role of new and current methods in semen technology for genetic resource conservation

2004 ◽  
Vol 30 ◽  
pp. 191-205
Author(s):  
W.V. Holt ◽  
P.F. Watson
Keyword(s):  
Reproductive Tract ◽  
Genetic Resource ◽  
Resource Conservation ◽  
Female Reproductive Tract ◽  
Practical Implementation ◽  
New Information ◽  
Frozen Semen ◽  
Genetic Resource Conservation

AbstractThe establishment of repositories of frozen semen, for the conservation of agricultural genetic resources, is not a simple matter of collecting and freezing semen in the hope that one day it will be suitable for use in an artificial insemination procedure. Important genetic issues need to be considered; for example, how many samples should be stored and from how many individuals? Aside from these, many biological and logistic issues must be considered. Cryopreservation technology does not work equally well in all species, often because of anatomical differences in the female reproductive tract leading to significant variability in the number of spermatozoa needed in order to achieve an acceptable conception rate. Moreover, spermatozoa from different species are not equally susceptible to cryoinjury. However, it is also emerging that semen samples from individuals within a species are also of different quality; several studies have revealed that these differences reflect the quality of DNA within the spermatozoon itself and also the efficacy of biochemical functions, including metabolic and signalling systems, within individual cells. As new possibilities to select spermatozoa for insemination arise, especially the use of flowsorting for gender selection, these issues may become more significant. In this article we interpret the way in which some of this new information may impact upon the practical implementation of genetic resource conservation.

Hydrogen peroxide levels in mouse oocytes and early cleavage stage embryos developed in vitro or in vivo

Development ◽  
1990 ◽  
Vol 109 (2) ◽  
pp. 501-507 ◽  
Author(s):  
M.H. Nasr-Esfahani ◽  
J.R. Aitken ◽  
M.H. Johnson
Keyword(s):  
Reproductive Tract ◽  
Female Reproductive Tract ◽  
Cell Stage ◽  
Mouse Oocytes ◽  
Early Embryos ◽  
Individual Mouse ◽  
The Relationship ◽  
Developmental Block

We describe a fluorimetric method for measuring the level of H2O2 in individual mouse oocytes and early embryos. Levels of H2O2 are low but detectable in unfertilized oocytes recovered freshly from the female reproductive tract. The levels in early cleaving embryos (1-cell to 8-cell stages) immediately after recovery from the female tract seem to be slightly higher the later the stage examined. However, when embryos are cultured in vitro from the 1-cell or early 2-cell stage, H2O2 levels rise when the embryos reach the mid-2-cell stage and remain elevated until they enter the early 4-cell stage. No equivalent elevation of H2O2 is seen during the transition from 1-cell to 2-cell or from 4-cell to 8-cell stages. Embryos that are able to develop successfully in vitro, as well as those that show a developmental block at the 2-cell stage on culture in vitro, both show this rise in H2O2 levels after in vitro culture. The relationship between the rise in H2O2 and the ‘2-cell block’ to development is discussed.

scholarly journals Effect of Ram Breed on the Efficiency of in vitroDevelopment of Sheep Embryos

2017 ◽  
Vol 14 (4) ◽  
pp. 1309-1313
Author(s):  
Y. Al-Anazi ◽  
M. G. Al-Mutary ◽  
M. M. Alfuraiji ◽  
M. Al-Ghadi ◽  
A. R. Al-himaidi ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Cell Stage ◽  
In Vitro Development ◽  
Progressive Motility ◽  
Frozen Semen ◽  
Stage 4 ◽  
Before And After ◽  
Vitro Development ◽  
Fresh Semen

The aim of this work was to investigate the impacts of ram breed on in vitro embryo development from fresh or frozen semen. Semen was collected from Najdi and Naimi rams and frozen; the mass and progressive motility of the spermwere assessed in each trial before and after freezing. Then, 970 oocytes in six replicates were fertilized with fresh and frozen semen in vitro. Different stages of sheep embryos were recorded. There were no significant differences in mass and progressive sperm motility of fresh or frozen ram semen between Najdi and Naimi,but there were significant differences between frozen and fresh semen within each breed. Our results showed significant (P<0.05) differences in 2-cell stage, 4-cell stage, 8-cell stage, morula, fragmented embryos, cleavage and blastocyst rates in the frozen semen group compared to fresh semen group in both breeds. In addition, significant (P<0.05) differencesbetween the two breeds were shown in 8-cell and16-cell embryonic stages.In conclusion, there were slight breed effects on the efficiency of in vitro development of sheep embryos.

Perineal swelling secondary to a vaginal wall cyst in a bitch in dioestrus

2019 ◽  
Vol 7 (3) ◽  
pp. e000805
Author(s):  
Cesar Sanchez Jimenez ◽  
Ana Rita Ramalho Furtado ◽  
Valéria Café Marçal ◽  
Darren Barnes ◽  
Emilie Fauchon
Keyword(s):  
Reproductive Tract ◽  
Periodic Acid ◽  
Vaginal Wall ◽  
Cystic Mass ◽  
Female Reproductive Tract ◽  
Intact Female ◽  
Periodic Acid Schiff ◽  
History Of ◽  
Perineal Swelling ◽  
Slow Growing

Vaginal cysts are a rare pathology of the female reproductive tract and are usually asymptomatic and often incidental in human and veterinary medicine. A seven-year and 11-month-old intact female bearded collie was referred for further investigations of a two-and-a-half-week history of a slow-growing perineal swelling that was noticed three to four weeks following oestrus. Ultrasound and CT images were acquired, which revealed that an intravaginal cystic mass was the cause of the aforementioned swelling. Following removal of the mass, histopathology revealed a fully excised vaginal cyst lined by simple cuboidal periodic acid-Schiff-positive epithelium most consistent with paramesonephric (Müllerian) origin. To the authors’ knowledge this is the first reported case of an extra-abdominal vaginal wall cyst in a bitch in dioestrus causing perineal swelling.

scholarly journals The Assessment of Longitudinal Sections of Rat Female Reproductive Tissues for NTP 2-Year Toxicity and Carcinogenicity Studies

2020 ◽  
Vol 48 (6) ◽  
pp. 747-755
Author(s):  
Susan A. Elmore ◽  
Chad Blystone ◽  
Beth A. Lubeck ◽  
Shawn F. Harris ◽  
Crystal L. Johnson
Keyword(s):  
Cross Sections ◽  
Reproductive Tract ◽  
Uterine Horn ◽  
Female Reproductive Tract ◽  
Uterine Tissue ◽  
Complete Evaluation ◽  
National Toxicology Program ◽  
Gross Lesions ◽  
Improved Accuracy ◽  
Tissue Origin

The National Toxicology Program (NTP) now uses an extended longitudinal sectioning protocol for the uterus to better evaluate female rodent reproductive tract toxicity for all developmental and reproductive toxicology and 2-year toxicity and carcinogenicity bioassays. The previous protocol for toxicity/carcinogenicity studies involved 1 cross section midway through each uterine horn and collection of uterine cervix and vagina only if gross lesions were present. Here we compare the histological findings of the original cross sections with the additional longitudinal sections of residual uterine tissues of 7 chronic NTP rat bioassays. The goal of this study was to determine whether there might be any advantages to examining additional uterine tissue. The longitudinal protocol allowed for 10 to 20 times more uterine tissue for evaluation. Results indicate that the potential advantages of a more complete evaluation of female reproductive tract tissue include increased detection of reproductive targets, increased detection of neoplastic and nonneoplastic lesions, improved detection of tissue origin of neoplasms, less reliance on gross identification of lesions, improved accuracy in the application of severity grades, and increased detection of preneoplastic or subtle lesions.

Effect of Contraceptive Steroids on the Endometrium of Guinea Pig: SEM study

1977 ◽  
Vol 35 ◽  
pp. 668-669
Author(s):  
Mai M. Said ◽  
Ramesh K. Nayak ◽  
Randall E. McCoy
Keyword(s):  
Guinea Pig ◽  
Reproductive Tract ◽  
Three Dimensional ◽  
Female Reproductive Tract ◽  
Human Female ◽  
Surface Ultrastructure ◽  
Transmission Electron ◽  
Sem Study ◽  
Uterine Mucosa ◽  
Group 1

Burgos and Wislocki described changes in the mucosa of the guinea pig uterus, cervix and vagina during the estrous cycle investigated by transmission electron microscopy. More recently, Moghissi and Reame reported the effects of progestational agents on the human female reproductive tract. They found drooping and shortening of cilia in norgestrel and norethindrone- treated endometria. To the best of our knowledge, no studies concerning the effects of mestranol and norethindrone given concurrently on the three-dimensional surface features on the uterine mucosa of the guinea pig have been reported. The purpose of this study was to determine the effect of mestranol and norethindrone on surface ultrastructure of guinea pig uterus by SEM.Seventy eight animals were used in this study. They were allocated into two groups. Group 1 (20 animals) was injected intramuscularly 0.1 ml vegetable oil and served as controls.

The application of electron microscopy to diagnosis in gynecologic neoplasms and tumor-like conditions

1984 ◽  
Vol 42 ◽  
pp. 102-105
Author(s):  
Lawrence M. Roth
Keyword(s):  
Electron Microscopy ◽  
Reproductive Tract ◽  
Malignant Tumors ◽  
Frozen Section ◽  
Cost Effective ◽  
Female Reproductive Tract ◽  
Ultrastructural Examination ◽  
Specific Diagnosis ◽  
Potential Value ◽  
Gynecologic Neoplasms

The female reproductive tract may be the site of a wide variety of benign and malignant tumors, as well as non-neoplastic tumor-like conditions, most of which can be diagnosed by light microscopic examination including special stains and more recently immunoperoxidase techniques. Nevertheless there are situations where ultrastructural examination can contribute substantially to an accurate and specific diagnosis. It is my opinion that electron microscopy can be of greatest benefit and is most cost effective when applied in conjunction with other methodologies. Thus, I have developed an approach which has proved useful for me and may have benefit for others. In cases where it is deemed of potential value, glutaraldehyde-fixed material is obtained at the time of frozen section or otherwise at operation. Coordination with the gynecologic oncologist is required in the latter situation. This material is processed and blocked and is available if a future need arises.

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