Evaluation of the 7 & 7 Synch and 7-day CO-Synch + CIDR treatment regimens for control of the estrous cycle among beef cows prior to fixed-time artificial insemination with conventional or sex-sorted semen.

An experiment was conducted to compare the 7 & 7 Synch and 7-day CO-Synch + controlled internal drug release (CIDR®) treatment regimens before fixed-time artificial insemination (FTAI) of beef cows with conventional or sex-sorted semen. Cows (n = 1538) were blocked based on age and days postpartum (DPP) and randomly assigned to treatment regimen and semen type. Cows assigned to the 7-day CO-Synch + CIDR treatment regimen (n = 769) were administered gonadotropin-releasing hormone (GnRH) and an intravaginal progesterone-releasing insert (CIDR) on Day - 10, and administration of prostaglandin F2α (PG) coincident with CIDR removal on Day - 3. Cows assigned to 7 & 7 Synch (n = 769) were administered PG and a CIDR device on Day - 17, GnRH on Day - 10, and PG coincident with CIDR removal on Day - 3. Cows were administered GnRH coincident with FTAI, which was performed 66 h after CIDR removal with conventional (20 × 106 cells) or sex-sorted (4 × 106 cells) semen. Expression of estrus was affected by treatment regimen (P = 0.01) and by treatment regimen × DPP (P = 0.0004), as a result of imposing the 7 & 7 Synch regimen; therefore, a greater percentage of cows expressed estrus (82% compared with 64%), particularly among cows with greater DPP. Pregnancy percentages resulting from FTAI were less (P < 0.0001) when using sex-sorted semen but greater among cows treated with 7 & 7 Synch (conventional semen: 72%; sex-sorted semen: 52%) compared with 7-day CO-Synch + CIDR (conventional semen: 61%; sex-sorted semen: 44%).

Development of the Timed Re-Insemination (TRI-synch) program re-inseminating 24 days after the initial service in dairy cows.

For the timed re-insemination at the minimal interbreeding interval, cows were treated with a progesterone (P4 )-releasing intravaginal device from Days 13-15 to 21 post-insemination (Day 0 = estrus), followed by plasma P4 assay on Day 23 and then subjected to the Experiments 1 and 2. In Experiment 1, of 18 cows, 6 cows were determined as luteolysis with low (<1 ng/ml) plasma P4 concentrations on Day 23 and ovulated on Days 24 (3 cows), 25 (1 cow), and 26 (1 cow) except a cow affected by ovarian quiescence. In Experiment 2, all cows were treated with GnRH on Day 23. Cows with low (<1 ng/ml) plasma P4 concentrations on Day 23 were diagnosed as non-pregnant and subjected to the re-insemination in the morning of Day 24 irrespective of estrous signs. Of 36 cows, 15 cows were diagnosed as being non-pregnant on Day 23. Fourteen cows of the non-pregnant animals were re-inseminated in the morning of Day 24 irrespective of estrous signs and the pregnancy rate of re-insemination was 36%. The conception rates of initial and re-inseminations were 50% (18/36) and 36% (5/14), respectively. The overall pregnancy rate by adding the rates of initial and re-inseminations was 64% (23/36).

An automated instrument for intrauterine insemination sperm preparation.

Sperm preparation is critical to achieving a successful intrauterine insemination and requires the processing of a semen sample to remove white blood cells, wash away seminal plasma, and reduce sample volume. We present an automated instrument capable of performing a sperm preparation starting with a diluted semen sample. We compare our device against a density gradient centrifugation by processing 0.5 mL portions of patient samples through each treatment. In 5 min of operating time, the instrument recovers an average of 86% of all sperm and 82% of progressively motile sperm from the original sample while removing white blood cells, replacing the seminal plasma, and reducing the volume of the sample to the clinically required level. In 25 min of operating time, density gradient centrifugation recovers an average of 33% of all sperm and 41% of progressively motile sperm. The automated instrument could improve access to IUI as a treatment option by allowing satellite doctor's offices to offer intrauterine insemination as an option for patients without the clinical support required by existing methods.

Cervical artificial insemination in sheep: sperm volume and concentration using an antiretrograde flow device.

There has been development of an antiretrograde flow device (DARIO), for sheep cervical artificial insemination (CAI). There, however, needs to be optimization of sperm volume and concentration of insemination doses when the DARIO is used for CAI. Objectives were to compare fertility rates (proportion of ewes lambing as a result of CAI) when there was use of the DARIO for CAI: two sperm volumes containing equal numbers of spermatozoa: 0.25 mL of 1,600 × 106 spermatozoa/mL and 0.50 mL of 800 × 106 spermatozoa/mL (Test 1 group), and two sperm volumes with a different number of spermatozoa/AI dose: 0.25 mL and 0.50 mL of 1,600 × 106 spermatozoa/mL (Test 2 group). There were 335 ewes from seven farms assigned to 60 batches (equally divided into a Control and Test 1 group). For the Test 2 group, 462 ewes from nine farms were assigned to 88 batches (equally proportioned into Control group and Test 2 groups). For the Test 1 group, proportion of ewes lambing as a result of CAI were 0.701 ±â€¯0.2679 and 0.595 ±â€¯0.2393 for the Control and Test 1 groups, respectively (P = 0.163). For the Test 2 group, proportions of ewes lambing were 0.550 ±â€¯0.2598 and 0.658 ±â€¯0.2412 for the Control and Test 2 group, respectively (P = 0.041). An inclusion of a larger number of spermatozoa per insemination in a 0.50 mL dose volume resulted improved proportion of ewes lambing as a result of CAI when there was used of the DARIO.

The impact of the modified speculum application technique on the success rates of intrauterine insemination: A randomized controlled study.

OBJECTIVE: Intrauterine insemination (IUI) is frequently used to treat patients with ovulation disorders, cervical factor, mild male infertility and unexplained infertility. The aim of this study was to investigate the impact of modified speculum application on the success of IUI in patients with unexplained infertility. MATERIALS AND METHODS: This prospective randomized study reviewed 219 women who had undergone controlled ovarian hyperstimulation (COH)-IUI treatment. In the modified speculum application group (109 patients with 124 cycles), the screw of the vaginal speculum was loosened after passing the internal os with catheter and the vaginal speculum remained in this position to ensure closure of the cervix during the procedure. In the conventional speculum application group (110 patients with 132 cycles), the screw of the vaginal speculum was not loosened to close the lips of cervix after passing the internal os with the catheter and the vaginal speculum was removed after withdrawal of the insemination catheter. The primary outcome was live birth rate. RESULTS: The modified and conventional speculum application groups had statistically similar demographic and clinical characteristics. There were no significant differences between the study and the control groups in terms of the clinical pregnancy rate per cycle and per patient (24.1% vs 18.9% and 26.6% vs 22.7%, respectively), as well as the live birth rate per cycle and per patient (19.3% vs 15.1% and 22% vs 18.1% respectively). CONCLUSION: Applying gentle mechanical pressure on the portio vaginalis of the cervix using a vaginal speculum during IUI does not improve pregnancy and live birth rates in patients with unexplained infertility.

Use of a sanitary sheath at artificial insemination by nonprofessional technicians does not markedly improve pregnancy rates to artificial insemination in pasture-based dairy cows.

Plastic sanitary sheaths over artificial insemination (AI) guns have been used at the time of AI to improve hygiene at AI and fertility in cattle, but fertility responses have been variable in studies when AI was performed by professional inseminators. The aims of this study were to investigate whether the use of a sanitary sheath at the time of AI carried out by nonprofessional (do-it-yourself, or DIY) inseminators improves pregnancy rates to AI in pasture-based dairy cows and whether effects of sheaths are greater in cows with contaminated vulvas and in cows at increased risk of extended calving to conception intervals. Lactating dairy cows located in 10 pasture-based herds in a subtropical region of northern Australia were inseminated by herd-based DIY inseminators and assigned to be inseminated with (n = 3,655) or without (n = 3,969) a sanitary sheath, with potential effects assessed using multivariable logistic regression. Overall, use of a sheath at the time of AI did not significantly affect pregnancy rates to AI (36.3% for those inseminated without a sheath vs. 36.8% for those inseminated with a sheath; odds ratio: 1.02; 95% confidence interval: 0.92-1.11). Effects of using a sheath on pregnancy rates to AI varied by herd, with lower pregnancy rates with the use of sheaths in 1 herd and some evidence of increases in 3 herds. Unexpectedly, there was evidence that the effect of sheath on pregnancy rates was less positive (or more negative) when the vulva was classified as dirty before any cleaning of the vulva before insemination compared with when the vulva was classified as clean (interaction odds ratio: 0.75; 95% CI: 0.56-1.00). Interactions between sheath and other explanatory variables that could affect fertility were not significant; thus, there was no compelling evidence that the effect of using a sheath was modified by any of these variables. We conclude that the use of sheaths during AI of pasture-based dairy cows by DIY inseminators does not, on average, markedly improve pregnancy rates to AI. However, responses may vary between herds, and the response to sheaths may be inferior (i.e., less positive or more negative) when a cow's vulva is contaminated with feces or discharge at the time of AI compared with when the vulva is clean.

Effect of post artificial insemination treatment with two different progesterone intravaginal devices on conception and synchronization of the returning estrus in Japanese Black cows.

The objective of this study was to evaluate the effects of post artificial insemination (AI) treatment with intravaginal progesterone device (P4 device) on conception rate, synchronization of returning estrus and plasma P4 concentration in Japanese Black cows. Nineteen cows were treated with DIB (1.0 g P4) from Day 12 to 19 (Day 0=day of the first AI), 27 cows were treated with a CIDR (1.9 g P4) from Day 12 to 19, and 33 cows were not treated after the first AI (control). Estrous behavior was daily examined between Day 20 and 25, and cows returning to estrus were inseminated (the second AI). On Day 19, plasma P4 concentration was not different among DIB, CIDR and control groups. There was no significant difference in conception rate after the first AI among three groups (DIB: 63.2%, CIDR: 66.7% and control: 72.7%). In non-pregnant cows, there was no significant difference in the proportion of cows showed returning estrus between Day 20 and 25 (DIB: 57.1%, CIDR: 22.2% and control: 44.4%), and day of returning estrus was not synchronized. The overall conception rate after the first and second AI was not different among the groups. In conclusion, post-AI treatment with intravaginal devices containing 1.0 and 1.9 g P4 from Day 12 to 19 neither increased plasma P4 concentration nor improved fertility and synchronization of the returning estrus in Japanese Black cows.

Morphometry and depth of inseminating catheter penetration in prolific and non- prolific ewes at different ages: A post mortem study.

The ewe's cervix structure is highly variable among females, which may limit the application of transcervical artificial insemination (TCAI). This work aimed to study the variation in cervix anatomy in two different prolificacy breeds (D'man a prolific and Boujaâd a non-prolific sheep). Reproductive tracts were collected from D'man (n = 64) and Boujaâd (n = 187) females aged between 1 and 4 years. The weight of the uterus, length of the cervix and depth of penetration of the inseminating pipette were recorded. The number of cervical rings was counted and their arrangement graded. The cervix anatomy of D'man ewes differs from that of Boujaâd. The cervix morphology of Boujaâd ewes is more complex compared to D'man ewes; this complexity depends on the age of the ewe. No significant difference was found regarding the depth of penetration of the insemination catheter for the two breeds at 1, 2 and 4 years old (P > 0.05). However, at 3 years old, the highest depth of penetration (P < 0.05) was recorded in D'man ewes. In addition, for both breeds, grade 2 was the most frequent (P < 0.05), and the cervix became less complicated for TCAI with advanced age (4 years old). To conclude, the cervix of D'man ewes is less complex and more favorable to TCAI. In addition, to overcome the complexity of the cervix in Boujaâd sheep, a selection of 4-year-old ewes will make the penetration of the artificial insemination catheter possible.

Comparison between an exclusive in vitro-produced embryo transfer system and artificial insemination for genetic, technical, and financial herd performance.

The objective of this study was to implement an in vitro-produced embryo transfer (IVP-ET) system in an existing stochastic dynamic dairy simulation model with multitrait genetics to evaluate the genetic, technical, and financial performance of a dairy herd implementing an exclusive IVP-ET or artificial insemination (AI) system. In the AI system, sexed semen was used on the genetically best heifers only. In the IVP-ET system, all of the animals in the herd were impregnated with female sexed embryos created through in vitro fertilization of oocytes collected from animals of superior genetics for different traits of interest. Each donor was assumed to yield on average 4.25 transferable embryos per collection. The remaining animals in the herd were used as recipients and received either a fresh embryo or a frozen embryo when fresh embryos were not available. Selection of donors was random or based on the greatest estimated breeding value (EBV) of lifetime net merit (NM$), milk yield, or daughter pregnancy rate. For both the IVP-ET and AI systems, culling of surplus heifer calves not needed to replace culled cows was based on the lowest EBV for the same traits. A herd of 1,000 milking cows was simulated 15 yr over time after the start of the IVP-ET system. The default cost to produce and transfer 1 embryo was set at $165. Prices of fresh embryos at which an exclusive IVP-ET system financially breaks even with the comparable AI system in yr 15 and for an investment period of 15 yr were also estimated. More surplus heifer calves were sold from the IVP-ET systems than from the comparable AI systems. The surplus calves from the IVP-ET systems were also genetically superior to the surplus calves from the comparable AI systems, which might be reflected in their market value as a premium price. The most profitable scenario among the 4 IVP-ET scenarios in yr 15 was the one in which NM$ was maximized in the herd. This scenario had an additional profit of $8/cow compared with a similar AI scenario that maximized NM$, provided that surplus heifer calves could be sold at a premium price based on the superiority of the EBV of NM$. For the IVP-ET system to be at least as profitable as the comparable AI system during a 15-yr investment period, the surplus calves from the IVP-ET system needed to be sold at the premium prices. The break-even price of fresh embryos was estimated to be $84 for the exclusive IVP-ET system. This resulted in the same profit as the AI system, which maximized NM$ for a 15-yr investment period and in which heifer calves were sold at a premium price.

Assessment of the usage of biodegradable polymeric matrix in vaginal devices to sustain progesterone release in cows.

The usage of timed artificial insemination (TAI) at a low cost leading to better reproductive rates has been the aim of several research groups in the field. Usually during TAI protocols, sustained progesterone (P4 ) release devices are employed. Most devices are constituted of a nylon skeleton covered with a silicon layer with P4 . A device based on biopolymers was developed in order to reduce costs and decrease its environmental impact. In this study, we compared the kinetics of sustained progesterone release among devices manufactured with a polymeric blend made of polyhydroxybutyrate-valerate (PHBV) and poly-ε-caprolactone (PCL) (DISP) which were compared with DIB® (Internal Bovine Device) used as the control. In the in vitro and in vivo progesterone release tests, two types of biopolymer-based devices with a superficial area of 147 cm2 were used: DISP8 (46% PHBV, 46% PCL and 8% P4 ; n = 4), DISP10 (45% PHBV, 45% PCL, 10% P4 ; n = 4) and DIB® (1 g P4 , 120 cm2 area; n = 3). The in vitro tests were carried out according to USP XXIII specifications and were performed in a dissolutor sink using an alcohol/water mixture (60/40 v/v) as a release media and samples were collected at 2 min, 2, 4, 8, 12, 24, 48, 60, 72, 84 and 96 h. P4 concentrations were measured through spectrophotometry in a 244 nm long wave. Three to 3 comparisons of angular coefficients of the straight lines obtained by regression analysis of accumulated P4 concentrations as a function of square root of time were carried out. Furthermore, the diffusion coefficient values of P4 were also determined for DISP8 and DISP10. The results showed that the concentrations of P4 were higher in the DISP10 (774.63 ± 45.26 µg/cm2 /t1/2 ) compared to DISP8 (566.17 ± 3.68 µg/cm2 /t1/2 ) (P < 0.05). However, both DISP10 and DISP8 P4 concentrations did not differ from DIB® (677.39 ± 16.13 µg/cm2 /t1/2 ). For the analysis of released quantities per day of the in vitro test, four periods were considered: 0-24, 24-48, 48-72 and 72-96 h. In the first 24 h, DISP8 released significantly less P4 than DISP10 or DIB®, which did not differ among them. Between 24 and 48 h, DISP10 released significantly more P4 than DIB®. DISP8 released an intermediate P4 amount and did not differ significantly from DIB® or DISP10. Between 48 and 72 h, P4 quantity released by DISP10 was significant higher (P < 0.01) than that of DIB® and DISP8, which did not differ among themselves. Between 72 and 96 h, DISP10 released significantly more P4 than DIB®, and DISP8 released an intermediate amount which did not differ from DIB® or DISP10 (P < 0.01). There was interaction between treatment and time (P = 0.0024). The diffusion coefficient values were: 1.36 × 10-8 (cm2 /s) for DISP10 and 1.12 × 10-8 (cm2 /s) for DISP8. For the in vivo test, ovariectomized crossbred cows received DIB® (n = 4) or DISP8 (n = 8) in an alternate design with a non-balanced sequence (cross-over) added of measures repeated in time referring to 16 days of blood samples collection. Samples were analyzed through radioimmunoassay in solid phase using the commercial kit of DPC (Diagnostics Products Corporation). Plasma concentrations of P4 peaked at 4 h after the placement of the device, this being the only time in which plasma P4 concentrations differed between DIB® (11.45 ± 1.96) compared with DISP8 (9.23 ± 1.15 ng/mL) (P = 0.027). On day 8, plasma P4 concentrations were similar for DIB® (2.44 ± 0.09) and DISP8 (1.89 ± 0.13 ng/mL) (P = 0.58) showing that both devices were able to keep P4 concentrations above 1 ng/mL in the plasma of the cow during the 16 day in vivo test. In conclusion, devices manufactured with the blend of PHBV/PCL biopolymers can sustain the release of P4 in a similar manner as silicon.

Esclerosis múltiple y embarazo: estudio unicéntrico prospectivo y comparativo / Multiple sclerosis and pregnancy: a single-centre prospective comparative study

Introducción: La esclerosis múltiple (EM) es una enfermedad autoinmune que afecta preferentemente a mujeres en edad fértil. Durante el embarazo y puerperio, cambia la tasa anual de brotes (TAB) de EM, sin modificar la evolución a largo plazo. Analizamos la repercusión del embarazo en pacientes con EM, y comparamos sus resultados obstétricos con embarazos de mujeres sanas. Métodos: Estudio unicéntrico, observacional descriptivo, de diseño longitudinal prospectivo. Se analizan los datos globales de una cohorte de pacientes con EM que han dado a luz entre enero de 2007 y julio de 2012, con un seguimiento de 2 años posparto. Los resultados obstétricos se compararon con un grupo control de 58 embarazadas sanas, elegidas al azar de nuestro centro durante el mismo período de tiempo. Resultados: Un total de 35 pacientes con EM, 40 partos. Grupo control: 58 mujeres, 60 partos. EDSS preembarazo: 0,7. TAB 2 años preembarazo: 0,5. Durante el embarazo: 0,3, a los 2 años posparto: 0,4. Doce pacientes recibían FME previo al embarazo, 4 iniciaron la gestación con FME. No hubo diferencias estadísticamente significativas en la TAB ni en la EDSS entre períodos preembarazo, embarazo y posparto. Al comparar con grupo control, no hubo diferencias en edad materna, semanas de gestación, peso al nacer, porcentaje de cesáreas, ni complicaciones obstétricas. En pacientes con EM hubo mayor porcentaje de tratamientos por infertilidad y menor porcentaje de lactancia, ambos estadísticamente significativos. Conclusiones: Nuestro trabajo confirma que el embarazo no repercute negativamente en el curso de la EM y que no existe mayor morbilidad obstétrica comparado con mujeres sanas

Introduction: Multiple sclerosis (MS) is a autoimmune disorder which preferentially affects young women of childbearing age. During pregnancy, the annualized relapse rate (AAR) is modified, but pregnancy has no harm effect on the long-term course of the disease. We aimed to study the clinical course of our MS patients during pregnancy, and compare their obstetrics outcomes with a control group of non-MS patients. Methods: A single centre prospective observational study was conducted. We assessed the reproductive history, MS history, pregnancy course and new-born outcome of a cohort of MS patients who had had a pregnancy between january 2007 and july 2012. We compared the global outcomes with a control cohort of 58 age-matched healthy pregnancies. Results: Complete data from 35 consecutive women were analyzed, 40 deliveries. Control groups: 58 patients, 60 deliveries. EDSS at pregnancy 0,7. ARR before pregnancy 0,5. During pregnancy 0,3, after pregnancy 0,4. Twelve patients were on disease-modifying drugs (DMD) before pregnancy, 4 prenatal exposure occurs. The comparison between relapse rate and EDSS before, during and after delivery showed no statistically significant difference. In addition, compared to control group, there were also no differences in the obstetric outcomes. In MS cohort, we found a higher incidence of assisted reproductive treatments and lower breastfeeding rate, both statistically significant. Conclusions: Our series confirms that pregnancy has no negative long term impact on the progression of MS and also suggest that there is no additional morbidity in the pregnancy, comparing to the rest of the population

The effect of slow release insemination on pregnancy rates: report of two randomized controlled pilot studies and meta-analysis.

PURPOSE: A modified application technique of intrauterine insemination (IUI) is slow release insemination (SRI), first described by Muharib et al. (Hum Reprod 7(2):227-229, 1992), who postulated higher pregnancy rates with a slow release of spermatozoa for 3 h. METHODS: To investigate this approach, two randomized controlled, cross-over pilot studies were performed from 2004 to 2006 in Israel and Germany to compare SRI with the standard bolus IUI. We aimed to present the results and perform a meta-analysis on available data for SRI. Univariate comparisons of pregnancy rates were performed using one-tailed z tests for method superiority. For meta-analysis, a fixed-effect Mantel-Haentzel weighted average of relative risk was performed. RESULTS: Fifty treatment cycles (IUI: n = 25, SRI: n = 25) were performed in Germany, achieving four pregnancies (IUI: 4%, SRI: 12%, p > 0.05). Thirty-nine treatment cycles (IUI: n = 19, SRI: n = 20) were performed in Israel achieving six pregnancies (IUI: 10.5%, SRI: 20%; p > 0.05). Meta-analysis of all eligible studies for SRI (n = 3) revealed a combined relative risk for pregnancy after SRI of 2.64 (95% CI 1.04-6.74), p = 0.02). CONCLUSIONS: In conclusion, these results lend support to the hypothesis that the pregnancy rate might be improved by SRI compared to the standard bolus technique.

Optimization of post-cervical artificial insemination in gilts: Effect of cervical relaxation procedures and catheter type.

Post-cervical (pC) artificial insemination (AI) has been successfully developed for application in multiparous sows, although it has proved problematic in gilts. This study analyzes the use of pC-AI in gilts by two experiments. In the first experiment, the efficiency of pC-AI in gilts was evaluated using a multi-ring multiparous catheter (MpC), which led to 23.1% of the gilts being successfully inseminated. In gilts where insemination was not possible using an MpC, two alternatives were applied before a second attempt at insemination: 1) Vetrabutin Chlorhydrate (VC) was intramuscularly injected in order to relax the cervix; or 2) Warm extender (WE) was deposited in the cervix to modify the cervical muscle dynamics. After the application of these treatments, the success rates achieved with the MpC were 34.2% and 23.8% for VC and WE, respectively. No statistically significant differences were found in the reproductive parameters measured [farrowing (%), litter size and fecundity index] between the use of MpC, or the MpC combined with VC or WE, compared with gilts inseminated by cervical AI (control group). In the second experiment, new catheters based on the anatomical characteristics of gilts (GpC) were used, and the rate of successful pC-AI application were compared (experiment 2a): a) MpC: control; b) GpC1: multi-ring catheter of Ø 16 mm and inner cannula of Ø 3.5 mm; c) GpC2: a multi-ring catheter of Ø with an inner cannula of Ø 2.5 mm. The highest rate of successful cannula penetration was reached in the GpC2 group (60.3%) followed by GpC1 (37.0%) and MpC (19.6%) (p < 0.05). There were no differences in the above mentioned reproductive parameters using the three catheters compared with cervical AI method (control group). Moreover, prior cervical AI did not improve subsequent pC-AI application 24 h later (experiment 2b). In conclusion, Vetrabutin Chlorhydrate, warm extender or the new catheters can be considered as useful tools for improving the success rate of pC-AI technique in gilts.

A new device to inseminate cows at the base of the uterine horns.

A new device (Chapingo device) to deposit semen at the base of the uterine horns of cattle was developed at Universidad Autonoma Chapingo, Mexico. Nine Holstein heifers were inseminated by transvaginal laparoscopy, using a laparoscope for cattle and the Chapingo device. A dose of sexed semen (2.1 × 106 spermatozoa) was deposited at the base of the uterine horn ipsilateral to the ovary where the preovulatory follicle was identified. Insemination was achieved in all the heifers, taking on average 13.7 ± 3.1 min per animal. In all cases, it was possible to see both ovaries, the base of the uterine horns and the oviducts. After the procedure, none of the heifers showed any type of complications such as haemorrhage, adhesions or trauma. On days 21 and 22 after insemination, four of the nine heifers (44.4%) returned into oestrus; on day 30 after insemination, one heifer was found to be pregnant by ultrasound. The results show the feasibility of generating pregnancies by transvaginal laparoscopy in heifers inseminated with sexed semen.

Technical Note: A new device for cervical insemination of sheep - design and field test.

Deep semen deposition, avoiding retrograde flow, lesions and stress, has proved to be very important in the success of sheep AI. The objective of the present study was to develop a new, suitable anti-retrograde flow device for sheep cervical AI (DARIO) that enables deep deposition of semen into the cervix without any modifications to the procedures currently used, and to compare the fertility, fecundity, and prolificacy rates between DARIO and a traditional catheter. Field tests were performed on 16 farms actively participating in the non-profit National Association of Rasa Aragonesa Breeders´ genetic selection scheme and where sheep management was similar. A total of 242 AI lots were considered, including 1,299 ewes; 126 lots (662 ewes) were inseminated using DARIO, and 116 lots (637 ewes) using a traditional commercially-available catheter (control group). Several factors affecting AI results were included in the model for mean comparison between DARIO and control groups (farm and ram as random factors; catheter, year and photoperiod as fixed effects; catheter × photoperiod interaction). The type of catheter had a significant effect on fertility ( < 0.01) and fecundity rates ( < 0.01) but no significant effect was detected on the prolificacy rate ( = 0.45). For fertility rate (percentage of ewes lambing after AI), means ± SE for DARIO and control groups were 59.44 ± 2.13% and 49.60 ± 2.48%, respectively; for fecundity rates, means ± SE for DARIO and control groups were 0.99 ± 0.04 and 0.82 ± 0.05 lambs/inseminated ewe, respectively, and, for prolificacy rates, means ± SE for DARIO and control groups were 1.68 ± 0.04 and 1.63 ± 0.04 lambs/ewe lambing, respectively. Fertility rate was greater in the decreasing photoperiod ( = 0.01). Significant effects were found for both year ( < 0.05) and farm ( < 0.01) on fertility, fecundity, and prolificacy rates. Neither ram nor catheter × photoperiod showed any significant effects on the variables investigated ( > 0.05). Overall, the use of DARIO instead of the traditional commercially-available catheter increased both fertility and fecundity rates; the marginal mean differences were 9.05 pregnant ewes per 100 inseminated and 0.15 lambs per inseminated ewe, respectively.

Development of a new device for artificial insemination in cynomolgus macaques.

In cynomolgus macaques, an important animal species for biomedical research, efficient reproduction has been hampered partly due to the difficulties of artificial insemination (AI) using straw tubes developed for humans or farm animals, because cynomolgus macaques have a complex cervical canal structure. In this study, taking into consideration the unique structure of the macaque cervical canal, we developed a novel device for AI, comprised of a syringe and an outer cylinder. At 24 and 48 h after using this device to inject semen into one female, viable sperm were observed in the oviduct where the sperm meets the oocytes. We then attempted AI using this new device on 10 females that were at pre-ovulation, and pregnancy was successful in three animals (30% pregnancy rate). These results show that the newly developed device can be used for AI in cynomolgus macaques.

Ovarian responses of dairy buffalo cows to timed artificial insemination protocol, using new or used progesterone devices, during the breeding season (autumn-winter).

This study evaluated the effect of new or used P4 devices on the ovarian responses of dairy buffalo that were administered an estradiol (E2) plus progesterone (P4)-based timed artificial insemination (TAI) protocol during the breeding season. On the first day of the TAI protocol, 142 cows were randomly assigned to receive one of the following: a new device (New; 1.0 g of P4; n = 48); a device that had previously been used for 9 days (Used1x, n = 47); or a device that had previously been used for 18 days (Used2x, n = 47). Ultrasound was used to evaluate the following: the presence of a corpus luteum (CL); the diameter of the dominant follicle (ØDF) during protocol; ovulatory response; and pregnancies per AI (P/AI). Despite similar responses among the treatments, there was a significant positive association of the ØDF during TAI protocol with ovulatory responses and number of pregnancies. In conclusion, satisfactory ovarian responses and a satisfactory pregnancy rate were achieved when grazing dairy buffalo were subjected to the TAI protocol in breeding season, independent of whether a new or used P4 device was used. Furthermore, the presence of the larger follicle was associated with a higher ovulation rate and higher P/AI following TAI.

A rapid and effective nonsurgical artificial insemination protocol using the NSET™ device for sperm transfer in mice without anesthesia.

Artificial insemination (AI) is an assisted reproductive technique that is implemented successfully in humans as a fertility treatment, performed extensively for commercial breeding of livestock, and is also successful in laboratory rodents. AI in the mouse may be especially useful for breeding of transgenic or mutant mice with fertility problems, expansion of mouse colonies, and as an alternative to in vitro fertilization. Nonsurgical AI techniques for the mouse have been described previously but are not often implemented due to technical difficulties. Here we compare various protocols for preparation of CD1 recipients prior to AI for naïve (in estrus), ovulation-induced, and superovulated females. Timing of hormone administration relative to sperm delivery is also compared. An improved protocol for nonsurgical AI in mice is described, which incorporates a convenient hormone administration schedule for female recipients and rapid, non-stressful sperm transfer without the need for anesthesia or analgesia.

Follicle and corpus luteum size and vascularity as predictors of fertility at the time of artificial insemination and embryo transfer in beef cattle / Tamanho e vascularização do folículo e corpo lúteo como preditores de fertilidade após inseminação artificial e transferência de embriões em gado de corte

Two ultrasound based fertility prediction methods were tested prior to embryo transfer (ET) and artificial insemination (AI) in cattle. Female bovines were submitted to estrous synchronization prior to ET and AI. Animals were scanned immediately before ET and AI procedure to target follicle and corpus luteum (CL) size and vascularity. In addition, inseminated animals were also scanned eleven days after insemination to target CL size and vascularity. All data was compared with fertility by using gestational diagnosis 35 days after ovulation. Prior to ET, CL vascularity showed a positive correlation with fertility, and no pregnancy occurred in animals with less than 40% of CL vascularity. Prior to AI and also eleven days after AI, no relationship with fertility was seen in all parameters analyzed (follicle and CL size and vascularity), and contrary, cows with CL vascularity greater than 70% exhibit lower fertility. In inseminated animals, follicle size and vascularity was positive related with CL size and vascularity, as shown by the presence of greater CL size and vascularity originated from follicle with also greater size and vascularity. This is the first time that ultrasound based fertility prediction methods were tested prior to ET and AI and showed an application in ET, but not in AI programs. Further studies are needed including hormone profile evaluation to improve conclusion.

Duas técnicas de predição de fertilidade, baseadas em ultrassonografia, foram testadas no momento da transferência de embriões (TE) e inseminação artificial (IA) em bovinos. Fêmeas bovinas foram submetidas a protocolos de sincronização de estro para TE e IA. Os animais foram escaneados por ultrassonografia imediatamente antes do procedimento de TE e IA para identificar o tamanho e vascularização do folículo e corpo lúteo (CL). Além disso, os animais inseminados foram escaneados onze dias após a inseminação para identificar o tamanho e vascularização do CL. Todos os dados foram comparados com a fertilidade utilizando-se do diagnóstico gestacional 35 dias após a ovulação. No momento da TE, a vascularização do CL apresentou-se positivamente relacionada com a fertilidade, sendo que animais com menos de 40% de vascularização do CL não ficaram gestantes. No momento da IA, assim como onze dias após a IA, nenhuma relação foi encontrada entre fertilidade e os parâmetros analizados (tamanho e vascularização do folículo e CL), enquanto que contrariamente, houve uma queda na fertilidade em vacas com vascularização do CL acima de 70%. Nos animais inseminados, o tamanho e vascularização do folículo foi positivamente relacionado ao tamanho e vascularização do CL, demonstrado pela presença de um CL maior e mais vascularizado proveniente de um folículo maior e mais vascularizado. O presente estudo é o primeiro a mostrar métodos de predição de fertilidade baseados em ultrassonografia no momento da IA e TE, demonstrando uma aplicabilidade prática no momento da TE. Novos estudos são necessários para suportar os resultados mostrados, incluindo análises hormonais.

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).