Uterine Body Insemination Research Articles

Comparison of efficiency between two artificial insemination methods using fresh semen in domestic cat (Felis Catus)

The objective of this study was to compare pregnancy rates in domestic cats using fresh semen for the bilateral intrauterine insemination (BIUI) method and the novel uterine body insemination (UBI) method. Queens received a single injection of equine chorionic gonadotropin (eCG) (200 IU; IM) to induce ovarian follicular development and, after 83 h, an injection of human chorionic gonadotropin (hCG) (100 IU; IM) for final oocyte maturation and ovulation induction. Thirty-four hours after hCG administration, 3 × 106 fresh spermatozoa were used for insemination by the BIUI (n = 8 queens) or by the UBI (n = 7 queens) techniques respectively. Pregnancy rates were 75.00% (6/8) by BIUI and 42.85% (3/7) by the UBI method. The mean litter size was 3.0 ± 0.86 for the BIUI, and 2.0 ± 1.0 for the UBI method. Spontaneous abortion occurred on day 35 of pregnancy in one queen following the UBI method. Our findings showed that the BIUI of queens with fresh semen resulted in higher pregnancy rates than the novel UBI method; also, acceptable pregnancy rates were achieved following BIUI with fresh semen in the domestic cat.

Sperm transport and endometrial inflammatory response in mares after artificial insemination with cryopreserved spermatozoa

This study aimed to determine whether the insemination site and dose with cryopreserved sperm of reproductively normal mares affect the sperm population in uterine tubes and the intensity of endometrial inflammatory response. Experimental subjects were estrous mares inseminated, in the mid-uterine body (Body) or the tip of the uterine horn (Tip), ipsilateral to the dominant follicle, with one 0.5 mL straw with 50 × 106 sperm (50) or with eight straws with 50 × 106 sperm/straw (400). Mares were slaughtered 2 h, 4 h and 12 h after artificial insemination (AI) and randomly assigned to following groups: Body 50 (n = 19) (2 h, 4 h or 12 h); Tip 50 (n = 29) (2 h, 4 h, or 12 h); Body 400 (n = 24) (2 h, 4 h, or 12 h); Tip 400 (n = 21) (2 h, 4 h, or 12 h). A Control group (n = 16) was not inseminated. After slaughter, uterine tubes were separated from uterus, and uteri and tubes flushed with phosphate-buffered saline (PBS). After flushing, an endometrial sample was collected from ipsilateral and contralateral horns and mid-uterus body for further histopathological examination. A sample of each uterine tube flushing was examined for sperm count, and a sample of each uterine flushing was used for polymorphonuclear neutrophils (PMNs) count. Data were analyzed using PROC GLM from SASv9.4. Insemination time, site, sperm dose, and their interactions were considered independent variables and sperm and PMNs numbers dependent variables. Deep horn insemination increased ipsilateral uterine tube sperm number without an increase in the inflammatory reaction compared with the uterine body insemination. The higher the insemination dose, the higher the uterine tubes’ sperm number and inflammatory reaction, with a quicker resolution. In conclusion, the insemination site and dose affected sperm in the uterine tubes, while post-insemination time and dose influenced the inflammatory reaction.

Inseminación artificial con semen congelado equino: reacción inflamatoria, transporte espermático y técnica de inseminación

This manuscript presents a brief historical review of investigations related to equine artificial insemination with frozen semen. The sperm-uterine interaction can be modified by the type of the insemination: concentration, total sperm numbers, motility/viability of sperm, volume, frequency and timing of AI. The frozen semen AI is widely used in the equine industry. The minimum recommended dose of spermatozoa has been established in 250 x 106 progressive motile sperm for uterine body insemination. However, the minimum dose required for maintaining the highest fertility for each stallion may differ greatly and therefore is determined by individual variation of intrinsic stallion fertility as well as with the insemination site. Deep intra-uterine insemination is commonly accepted as a routine procedure for AI with low doses. The number of AIs per cycles does not seem to be a factor affecting the final outcome of pregnancy as long as the last AI is performed within 12 h before and 12 h after the ovulation. Contrary to the common belief, the mare’s oocyte remains viable and fully capable of fertilization for 12-15 h, and the embryo loss rate after post-ovulatory insemination does not begin to increase until 12 h post-ovulation. Therefore, in a postovulatory insemination regime, examination of mares at intervals of any less than 12 h does not improve pregnancy or embryo loss rates.

Effects of Hysteroscopic and Uterine Body Insemination on the Presence of Selected Immune Cells in the Equine Endometrium

The effects of standard uterine body and hysteroscopic insemination on endometrial health were investigated. For this purpose, 33 mares were assigned to five different protocols: control (no insemination; n = 7), sham AI (sham uterine body insemination; n = 6), sham HysAI (sham hysteroscopic insemination; n = 7), standard AI (standard uterine body insemination, 300 × 106 progressively motile sperms (PMS); n = 7) and HysAI (hysteroscopic insemination, 100 × 106 PMS; n = 6). Sampling included uterine swabbing for microbiological examination, cytology for determination of polymorphonuclear neutrophils (PMNs) in the uterus, and endometrial biopsy collection for histology and characterization of endometrial immune cells on day 18 after ovulation (B1) as well as 8–10 hours (B2, day 20) and 72 hours after insemination (B3, day 23). Microbial contamination increased throughout the experiment in the sham insemination groups. Significant effects (P < .05) over time were detected for PMNs (cytology: sham HysAI, standard AI, and HysAI; histology: standard AI and HysAI), macrophages (immunohistochemistry: standard AI and HysAI) and T cells (immunohistochemistry: standard AI), showing an increase at B2 and a subsequent decrease toward baseline levels at B3. At B2, significant differences (P < .05) existed for PMNs (mean ± SEM) between control (1.3 ± 1.9%) and sham AI (2.2 ± 2.7%) versus standard AI (12.2 ± 4.7%) and for macrophages between control (4.1 ± 3.5%) and sham AI (2.5 ± 1.3%) versus standard AI (25.4 ± 15.8%). Thus, the cellular immune response of the endometrium depends on sperm deposition in the uterus and does not differ between hysteroscopic and standard uterine body insemination.

Effect of artificial insemination protocol and dose of frozen/thawed stallion semen on pregnancy results in mares.

Deep intra-uterine insemination is commonly accepted as a routine procedure for artificial insemination in horses. The motives and principles of deep insemination are well described, but the equipment used may differ. In this trial, the efficiency of two different insemination pipettes for deep intra-uterine insemination in the mare was compared with insemination into the uterine body using commercially available frozen-thawed semen of two stallions of proven fertility. These inseminations were performed using two different doses. The semi-flexible Minitube pipette was compared with a newly designed insemination device with a more flexible telescopic insemination catheter (Ghent device). The semi-flexible Minitube pipette performed better than the newly designed insemination device with respect to pregnancy outcome (p=0.008). The superiority of deep horn insemination over uterine body insemination was reflected by the better pregnancy rates obtained after deep insemination using the same low doses (30.6% better pregnancy rates) (p=0.0123).

Persistent Breeding‐Induced Endometritis after Hysteroscopic Insemination in the Mare

Low-dose insemination has been proposed to reduce persistent breeding-induced endometritis (PBIE) in mares with delayed uterine clearance (DUC). Others proposed that hysteroscopic insemination induces an exaggerated inflammatory response and should be avoided in DUC mares. The objectives here were to evaluate presence and severity of PBIE in normal and DUC mares after hysteroscopic insemination with fresh semen, and to determine if hysteroscopy could be used in DUC mares without inducing excessive inflammation. Reproductively normal (n = 4) and DUC (n = 5) mares received four treatments in random order: uterine body insemination (UB, 1 × 10(9) spermatozoa, 20 ml), hysteroscopic insemination (HYST, 5 × 10(6) spermatozoa, 0.5 ml), sham hysteroscopic insemination (SHAM, semen extender, 0.5 ml) and hysteroscopic infusion of seminal plasma (SP, 0.5 ml). Significantly more DUC (50%) mares than normal (14%) mares accumulated intrauterine fluid 24 h post-treatment. The difference in fluid accumulation between DUC (40%) mares and normal (7%) mares was also significant 48 h post-treatment. Fluid scores were not significantly different between treatments in normal mares. However, treatments HYST and SHAM resulted in significantly higher fluid scores 24 h but not 48 h post-treatment in DUC mares. There was no effect of treatment or mare group on the percentage and total number of neutrophils in uterine fluid 48 h post-treatment. Percentage of neutrophils was correlated with duration of hysteroscopy in normal mares, with procedures lasting ≥ 9 min associated with PBIE. There was no effect of mare group, treatment or duration of hysteroscopy on pregnancy rate. Hysteroscopy induces a transient inflammation that is not more severe than that after conventional artificial insemination, suggesting no contraindication to its use in DUC mares.

Effect of semen deposition site on conception rate in dairy cows

The aim of this study was to determine the efficiency of intracornual insemination versus uterine body insemination using a standard insemination dose. All cows have synchronised oestrus with Ovsynch protocol and only cows with strong oestrus signs were inseminated. An experiment was conducted in autumn. In group I 36 cows were inseminated into the uterine body and in group II 33 cows were inseminated into the uterine horn ipsilateral to the ovary with the largest follicle determined by ultrasonography. The conception rate was 69.44% in group I and 69.66% in group II (there were no significant differences). It was concluded that in routine insemination practice the site of semen deposition (into uterine body or into ipsilateral horn) does not play a significant role.

Pregnancy rates in cattle with cryopreserved sexed sperm: Effects of sperm numbers per inseminate and site of sperm deposition

In six field trials, doses between 1.0 and 6.0 × 10 6 total sexed, frozen-thawed sperm were inseminated into the uterine body or bilaterally into the uterine horns of heifers and nursing Angus cows 12 or 24 h after observed estrus. Except for one comparison in one trial in which uterine body insemination was slightly superior ( P < 0.05) to uterine horn insemination, there was no significant ( P > 0.1) difference between sites of semen deposition. Additionally, except for one small study with limited numbers, there was essentially no difference in pregnancy rates in the range between 1.5 and 6 × 10 6 sexed, frozen-thawed sperm per inseminate. Pregnancy rates with smaller doses of sexed sperm averaged about 75% of controls of 20 × 10 6 total frozen-thawed, unsexed sperm. While 1.0 × 10 6 sexed, frozen-thawed sperm per insemination dose resulted in decreased pregnancy rates compared to larger doses, the lesser fertility with sexed sperm could not be compensated by increasing sperm numbers in the range of 1.5–6 × 10 6 sperm per dose. Pregnancy rates with 2 × 10 6 sexed, frozen-thawed sperm per dose were not markedly less than control pregnancy rates with 20 × 10 6 frozen-thawed unsexed sperm/dose in well-managed herds.

Hysteroscopic insemination of small numbers of spermatozoa at the uterotubal junction of preovulatory mares

Mares were inseminated with motile spermatozoa suspended in 30-150 microliters Tyrode's medium directly onto the uterotubal papilla at the anterior tip of the uterine horn, ipsilateral to the ovary containing a dominant preovulatory follicle of > or = 35 mm in diameter, by means of a fine gamete intrafallopian transfer (GIFT) catheter passed through the working channel of a strobed light videoendoscope. Insemination of 10, 8, 25, 14, 11 and 10 mares with, respectively, 10.0, 5.0, 1.0, 0.5, 0.1 or 0.001 x 10(6) motile spermatozoa resulted in conception rates of, respectively, 60, 75, 64, 29, 22 and 10%. Deposition of 1.0 x 10(6) motile spermatozoa onto the uterotubal papilla began to approach the limit of successful fertilization. These doses are far lower than the 3-15 x 10(9) spermatozoa normally ejaculated by fertile stallions during mating, and the accepted minimum dose of 500 x 10(6) spermatozoa used for conventional uterine body insemination in mares. The simplicity of the technique offers a practical means of exploiting new breeding technologies that require very small numbers of spermatozoa in horse breeding.

Effect of a deep uterine insemination on spermatozoal accessibility to the ovum in cattle: A competitive insemination study

A competitive insemination study was conducted to determine the effect of a deep uterine insemination on accessory sperm number per embryo in cattle. Cryopreserved semen of a fertile bull characterized by spermatozoa with a semi-flattened region of the anterior sperm head (marked bull) was matched with cryopreserved semen from an unmarked bull having spermatozoa with a conventional head shape. Using 0.25-mL French straws and a side delivery embryo transfer device, deep uterine insemination (0.125 mL deposited in each horn) was performed 2 cm from the uterotubal junction. Immediately after, the uterine body was artificially inseminated using semen (0.25 mL) from an alternate bull and a conventional insemination device. The complete dose (both inseminations) was 50 × 10 6 total sperm cells consisting of an equal number of spermatozoa from each bull. Single ovulating cows (n = 95) were inseminated at random with either the unmarked semen in the uterine body and marked semen in the uterine horn, or the unmarked semen in the uterine horn and marked semen in the uterine body. Sixty-one embryos(ova) were recovered nonsurgically 6 d post insemination, of which 40 were fertilized and contained accessory spermatozoa. The ratio and total number of accessory spermatozoa recovered was different among treatments: 62:38 (326) for the unmarked semen in the uterine body and marked semen in the uterine horn, and 72:28 (454) for the unmarked semen in the uterine horn and marked semen in the uterine body (P<0.05). Deep uterine insemination using this semen in a split dose and a side delivery device favors accessibility of spermatozoa to the ovum compared with conventional uterine body insemination.

Impact of Site of Inseminate Deposition and Environmental Factors That Influence Reproduction of Dairy Cattle

Uterine body and cornual inseminations (n = 2127) were evaluated over a 3-yr period in Holstein and Jersey cattle. For cornual insemination one-half of each semen dose was deposited approximately 2.5cm into each uterine horn. Conception rate was lower for cervical insemination (39.4%) than uterine body (48.1%) or cornual (49.3%) inseminations. Pregnancy site distribution was equal for both insemination techniques but cervical insemination resulted in 60% right horn pregnancies. Primary housing, service number, inseminator, lactation number, and sire affected conception rate. Older cows were least fertile (31.4%), second service conception rate was lowest (42.6%), and barn housed cattle had a 39.7% conception rate. Days open was affected by primary housing, service number, sire, site of semen placement, and twinning. Twinning increased days open by 10 d. Optimum time for insemination of lactating cows was between 6 and 12h after the initial observation of estrus. From this study we conclude that shallow cornual insemination is as effective as uterine body insemination, and conception rate is optimized when estrus is positively assessed.

Increase of pregnancy rate in dairy cattle after preovulatory follicle palpation and deep cornual insemination

A total of 334 first-service lactating cows in natural estrus were used in the study. Semen was deposited into the uterine body of 174 cows and deep into the uterine horn ipsilateral to the side of impending ovulation of 160 cows. In both groups, insemination was performed within the interval of 50 to 100 d postpartum at 8 to 15 h after estrus detection and after preovulatory follicle palpation. Pregnancy rates were determined by palpation per rectum 50 d post insemination. The pregnancy rate was higher (P < 0.05) for deep uterine horn insemination (70.62%) than for uterine body insemination (60.34%).

Influence of cornual insemination on endometrial damage and microbial flora in the bovine uterus.

The objectives were to determine effects of cornual insemination on endometrial characteristics and uterine bacterial flora. Forty cows were assigned randomly (10/group) to each of the following treatments: 1) control--no uterine manipulation; 2) uterine body insemination--deposition of the inseminate at the internal cervical os; 3) cornual insemination--one-half of inseminate deposited into each uterine horn anterior to the internal bifurcation and 4) cornual insemination plus endometrial biopsy--insemination into each uterine horn and trauma induced by endometrial biopsy. Animals were slaughtered and reproductive tracts were removed 5 d after treatment. Bacterial populations of the uterine lumen were evaluated by both flushings and swabbings of each horn. Treatment did not influence the presence of luminal bacteria as determined by flushing or swab samples. Gross endometrial characteristics were quantitated in all tracts. Characteristics considered were total endometrial surface area, discolored regions that appeared to be distinctly darker than the remaining portion of the endometrium and obvious surface damage. Discolored regions were evaluated as both total areas and percentage of endometrium. Treatment did not influence total or percent discolored areas. Microscopic evaluation of these tissues revealed increased edema when compared with nondiscolored regions but no disruption of the luminal epithelium was found. Surface damage was found only in the biopsied treatment group. Observation of the endometrium revealed that extensive regeneration occurred by 5 d after treatment. The data suggest that cornual insemination does not result in uterine trauma or bacterial presence 5 d after breeding, and the endometrium is capable of regeneration after trauma during estrus.