Investigation of in vitro measurable sperm attributes and their influence on electroejaculated bull semen with a fixed‐time artificial insemination protocol in Australian Bos indicus cattle

Increasing use of fixed‐time artificial insemination (FTAI) in beef cattle production has presented an opportunity for the use of fresh or chilled semen as an alternative to standard cryopreserved semen. The objective of this study was to examine in vitro sperm function and pregnancy rate of electroejaculated semen, chilled and stored for 48 hr, compared to conventionally cryopreserved semen with an optimized FTAI protocol in Brahman cattle. Semen from three Brahman bulls was collected, and aliquots were extended in either chilled (at 5°C) or frozen (LN₂) in a Tris‐egg yolk extender base with 2.4% or 7.0% glycerol, respectively. Semen samples were assessed 48 hr after collection or post‐thaw and warming, for sperm motility, in vitro sperm function and fertilizing ability, and used in a FTAI programme. The overall pregnancy rates was significantly different (p < .01) after FTAI with frozen (n = 173; 53.2%) and chilled semen (n = 174; 31.6%). In contrast, the in vitro sperm assessment showed that the chilled semen had significantly faster motility (p < .05), a higher proportion of progressively motile spermatozoa (p < .05), with significantly higher proportions of acrosome intact, viable spermatozoa (p < .01). This study showed that reasonable pregnancy rates in Brahman cattle can be achieved using FTAI with chilled semen collected using electroejaculation and stored for up to 48 hr. However, improvements in semen extenders are required in consideration of semen collection method to improve the longevity of sperm fertilizing ability to significantly increase FTAI output using chilled storage of bull semen.     

ACE inhibition in goats under fixed‐time artificial insemination protocol increases the pregnancy rate and twin births

To assess the effect of the angiotensin‐converting enzyme (ACE) inhibition on the efficiency of the fixed‐time artificial insemination (TAI), 69 goats were divided randomly into two groups: enalapril (n = 35) and control (n = 34). In the experiment, all animals underwent the protocol of fixed‐time artificial insemination for 12 days. Enalapril group received enalapril maleate dissolved in saline (Enalapril, Lab Teuto Ltda) subcutaneously at the following doses: 0.2 mg/kg/day in D0‐D2; 0.3 mg/kg/day in D3‐D6 and 0.4 mg/kg/day in D7‐D11. The control group received the corresponding volume of 0.9% saline solution. We performed a single insemination 36 hr after sponge removal using frozen semen from two adult male goats with recognized fertility. The ultrasound pregnancy diagnosis was 30 days after the artificial insemination (AI). There was significant increase in pregnancy rates and twinning as well as a decrease in foetal loss in animals receiving enalapril (p < .01). The use of ACE inhibitors during the TAI protocol was shown to be a promising alternative to increase the efficiency of such reproductive biotechnology.     

Fertility of lactating dairy cows inseminated with sex‐sorted or conventional semen after Ovsynch,Presynch–Ovsynch and Double‐Ovsynch protocols

The objective was to compare pregnancy per artificial insemination (P/AI) with conventional (CS) or sex‐sorted semen (SS) in dairy cows subjected to one of the three timed AI protocols. Cows (n = 356) were randomly assigned to synchronization with Ovsynch (OVS), Presynch–Ovsynch (PO) or Double‐Ovsynch (DO) and inseminated on Day 77 ± 3 postpartum with either frozen‐thawed SS (n = 182) or CS (n = 184) of the same bull. More cows were cyclic at the beginning of breeding Ovsynch increased (p < 0.01) with presynchronization and it was greater for DO than PO (OVS = 78.5%, PO = 85.1%, DO = 95.6%). Overall, P/AI for SS and CS increased with presynchronization (p < 0.05) on Days 31 (OVS = 35.5%, PO = 47.1%, DO = 48.3%) and 62 (OVS = 30.1%, PO = 43.8%, DO = 43.9%). Regardless of synchronization treatments, insemination with SS reduced P/AI (p < 0.02) on Days 31 (38.1% vs. 50.6%) and 62 (34.5% vs. 45.6%) compared with CS. No interaction was observed between synchronization treatment and type of semen for P/AI, although in cows receiving CS, P/AI was numerically greatest for PO (OVS = 42.0%, PO = 59.3%, DO = 49.0%), and in cows receiving SS, it was numerically greatest for those inseminated following DO (OVS = 27.9%, PO = 35.5%, DO = 47.6%). Thus, presynchronization improved P/AI in cows inseminated with sex‐sorted or conventional semen.     

Effects of supplementation of iodixanol to semen extender on quality and fertilization ability of frozen–thawed Thai native bull sperm

This study investigates the effects of iodixanol supplementation in varied concentrations to Tris egg yolk (TEY) extender on the quality and fertilization ability of frozen–thawed sperm of Thai native bulls. Each ejaculate was divided into four different groups, as follows: sperm were treated with TEY extender (control group) and TEY extender supplemented with three different concentrations of iodixanol (1.25%, 2.50% and 5.00%). Semen straws were frozen in liquid nitrogen vapor. After thawing, sperm motility characteristics, viability, plasma membrane integrity and acrosome integrity were determined. Also, frozen–thawed spermatozoa from all groups were used for in vitro fertilization and artificial insemination (AI) in natural estrus Thai native cows. The results showed that the post‐thaw quality of the 2.50% iodixanol group was superior to the other iodixanol groups (P < 0.05). However, iodixanol had no beneficial effect on post‐thaw sperm in vitro fertilization ability and pregnancy rate after AI (P > 0.05). It can be concluded that the supplementation of 2.50% iodixanol extender significantly improves the progressive motility, viability, plasma membrane integrity and acrosome integrity of cryopreserved semen from Thai native bulls, but it has no beneficial effect on in vitro fertilization ability and pregnancy rate after AI.     

Combined cryopreservation of canine ejaculates collected at a one-hour interval increases semen doses for artificial insemination without negative effects on post-thaw sperm characteristics

A limiting factor in canine artificial insemination (AI) is the low number of insemination doses obtained per ejaculate. In this study, semen was collected from dogs (n = 28) either once and frozen directly after collection or the same dogs were submitted to a dual semen collection with a 1-hr interval and the two ejaculates were combined for cryopreservation. We hypothesized that combining two ejaculates increases semen doses per cryopreservation process without negative effects on semen characteristics. Total sperm count was lower in semen from a single semen collection in comparison with the combination of the first and second ejaculate of a dual semen collection (p < .001). The percentage of motile and membrane-intact spermatozoa determined by computer-assisted sperm analysis (CASA) in raw semen did not differ between single and combined dual ejaculates and was reduced (p < .001) by cryopreservation to the same extent in single (motility 73.7 ± 1.8%, membrane integrity 65.6 ± 2.2%) and combined dual ejaculates (motility 72.7 ± 2.3%, membrane integrity 64.6 ± 2.5%). The percentage of spermatozoa with morphological defects increased after cryopreservation (p < .001) but was similar in single and combined dual ejaculates. The CASA sperm velocity parameters decreased with cryopreservation (p < .001) but did not differ between single and combined dual ejaculates. The number of insemination doses increased from 2.7 ± 0.4 for single to 4.7 ± 0.8 for combined dual ejaculates (p < .01), based on 100 million motile spermatozoa per frozen-thawed semen dose. In conclusion, combining two ejaculates collected at short interval for one cryopreservation process increases the number of AI doses without compromising semen quality.     

Addition of oxytocin to semen extender improves both sperm transport to the oviduct and conception rates in pigs following AI

Oxytocin (OXT) contained in boar semen is known to produce uterine contraction; therefore, we hypothesized that the co‐injection of OXT with sperm would improve artificial insemination (AI) using liquid or frozen‐thawed boar sperm. We initially examined whether OXT added to semen extender improved sperm transport to the oviduct. Although the addition of OXT did not affect the fresh or frozen‐thawed sperm motility or acrosomal integrity, it significantly increased the number of sperm in the oviduct at 6 h after AI injection with OXT, as compared with the control (P < 0.05). Moreover, some sperm were observed in the sperm reservoir of the isthmus in the OXT treatment group, whereas few sperm were observed in the control. When OXT was added to the semen extender immediately prior to AI, the conception rates were significantly higher in both fresh semen and frozen‐thawed semen than in the control group (P < 0.05: liquid, 87.5% vs. 70.5%; frozen‐thawed, 89.8% vs. 75.0%). From these results, we concluded that the addition of OXT to the semen extender assisted in sperm transportation from the uterus to the oviduct, which resulted in improved reproductive performance.     

Timed artificial insemination in crossbred mares: Reproductive efficiency and costs

Timed artificial insemination (TAI) has boosted the use of conventional artificial insemination (CAI) by employing hormonal protocols to synchronize oestrus and ovulation. This study aimed to evaluate the efficiency of a hormonal protocol for TAI in mares, based on a combination of progesterone releasing intravaginal device (PRID), prostaglandin (PGF_2α) and human chorionic gonadotropin (hCG); and compare financial costs between CAI and TAI. Twenty-one mares were divided into two groups: CAI group (CAIG; n = 6 mares; 17 oestrous cycles) and TAI group (TAIG; n = 15 mares; 15 oestrous cycles). The CAIG was subjected to CAI, involving follicular dynamics and uterine oedema monitoring with ultrasound examinations (US), and administration of hCG (1,600 IU) when the dominant follicle (DF) diameter's ≥35 mm + uterine oedema + cervix opening. The AI was performed with fresh semen (500 × 10⁶ cells), and embryo was recovered on day 8 (D8) after ovulation. In TAI, mares received 1.9 g PRID on D0. On D10, PRID was removed and 6.71 mg dinoprost tromethamine was administered. Ovulation was induced on D14 (1,600 IU of hCG) regardless of the DF diameter's, and AI was performed with fresh semen (500 × 10⁶ cells). On D30 after AI, pregnancy was confirmed by US. The pregnancy rate was 80.0% in TAIG and 82.3% in CAIG (p > .05). The TAI protocol resulted in 65% reduction in professional transport costs, and 40% reduction in material costs. The TAI was as efficient as CAI, provided reduction in costs and handlings, and is recommended in mares.     

Fertility of Holstein cows and heifers submitted to timed artificial insemination and receiving one or two doses (12 h apart) of semen

The objective of this retrospective study was to assess the effect of receiving a single (n = 50,285) or double (n = 4392) artificial insemination (AI), 12 h apart, within a timed artificial insemination protocol on pregnancy per AI (P/AI) in nulliparous heifers (inseminated with either sex-sorted or conventional semen) and pluriparous Holstein cows in a commercial dairy herd. Also, this study aimed to investigate the relationship between temperature-humidity index (THI) and time of the first AI and fertility. Fertility of cows receiving two AI with normothermia (THI <68) was higher (p < .05) than cows receiving a single AI (42.9% vs. 36.4%). P/AI of cows receiving two AI with severe heat stress (THI >85) was higher (p < .05) than cows receiving a single AI (21.0% vs. 12.6%). Regardless of heat stress conditions, applying the first AI in the morning increased (p < .05) P/AI in cows with double AI than in cows whose first AI occurred in the afternoon (38.4 vs. 33.3%). With moderate heat stress, and sexed-sorted semen, P/AI to timed AI was higher (65.0 vs. 51.9%; p < .05) in heifers receiving double AI than those serviced once. It was concluded that double AI, 12 h apart, enhanced fertility at timed AI than herd mates with a single AI, particularly with heat stress at breeding.     

Effect of delaying the time of insemination with sex-sorted semen on pregnancy rate in Holstein heifers

The objective of the study was to evaluate the interval from onset of oestrus to time of artificial insemination (AI) to obtain the optimum pregnancy rate with sex-sorted semen in Holstein heifers. Heifers in oestrus were detected and inseminated only by using heat–rumination neck collar comprised electronic identification tag at the age of 13–14 months. Heifers (n = 283) were randomly assigned to one of three groups according to the timing of insemination at 12–16 hr (G1, n = 97), at 16.1–20 hr (G2, n = 94) and at 20.1–24 hr (G3, n = 92) after reaching the activity threshold. The mean duration of oestrus was 18.6 ± 0.1 hr, and mean peak activity was found at 7.5 ± 0.1 hr after activity threshold. The mean interval from activity threshold to ovulation was 29.4 ± 0.4 hr. The overall pregnancy per AI (P/AI) was 53.0% at 29–35 days and 50.9% at 60–66 days after AI. There was a significant reduction between G1 (13.8 ± 1.4 hr) and G3 (7.9 ± 1.4 hr) related to the intervals from AI to ovulation time. Sex-sorted semen resulted in significantly higher P/AI at 29–35 days when heifers inseminated in G3 (60.9%) after oestrus than those inseminated in G1 (49.5%) and G2 (48.9%). In terms of fertility, when the temperature–humidity index (THI) was below the threshold value (THI ≤65) at the time of AI, there was a tendency (≤65; 57.2% vs. > 65; 47.1%) for high pregnancy rate. There was no effect of sire on P/AI. In addition, the interaction of the technician with the time of AI was found significant, and three-way interaction of technician, sire and time of AI was tended to be significant on pregnancy rate. Thus, in addition to delaying the time of insemination (between 20.1 and 24 hr) after oestrous detection, THI and experienced technician were also found to be critical factors in increasing fertility with the use of sex-sorted semen in Holstein heifers.     

Influence of constant long days on ejaculate parameters of rabbits reared under natural environment conditions of Mediterranean area

Since rabbit bucks are usually housed under constant long daylight in artificial insemination (AI) centers, the main purpose of this study was to investigate whether constant long day influenced ejaculate parameters of rabbits housed in AI centers in the Spanish Mediterranean area. The study was carried out in Murcia, Spain (37° N). Twenty commercial hybrid male rabbits, aged between 14 and 15 weeks, were randomly allotted to two groups and housed under either natural day length (n=10, ND) or a constant 16-h daylight exposure of 16 h (n=10, CLD). Other management conditions, such as air temperature or reproductive handling, were identical for both groups. Two successive ejaculates were collected twice weekly from every male, and the first one was used to monitor ejaculate characteristics. Measurement of semen production, in terms of ejaculate and semen volume, sperm concentration and total sperm per ejaculate, and sperm quality, in terms of motility index, viability, morphology and acrosome integrity, was assessed in 783 ejaculates collected during 15 months (from October to December). No differences (P>0.05) in either semen production or sperm quality were shown among ejaculates collected from rabbits housed under ND and CLD conditions. A limited influence of season was observed (P<0.01); semen volume and motility index were highest and lowest, respectively, during summer. The increase of air temperature and humidity index (THI) had a significant detrimental effect (P<0.01) on both sperm production and quality parameters with a lag of 6 and 3 weeks, respectively. On the basis of these findings, annual variations of semen production and sperm quality in male rabbits seems more related to THI than to daylight length under conditions of AI management in the Mediterranean area of Spain.     

Pregnancy and offspring sex ratio following insemination with SexedULTRA and conventional semen in cows in a commercial beef operation

Pregnancy rate per AI (PR/AI) and breeding season pregnancy rates between insemination with sexed semen (SS; at 18 hr after the onset of oestrus) and conventional semen (CS; at 12 hr after the onset of oestrus,) and offspring gender ratio between two groups were compared. Angus cross cows (n = 686, during 2019 and 2020 breeding seasons) were oestrus-synchronized using Select-Synch + CIDR protocol and were observed thrice daily for oestrus until 72 hr after PGF2α administration. Cows expressed oestrus (n = 513) were inseminated with either SS (n = 246; SexedULTRA 4M™; y chromosome-bearing sperm) or CS (n = 267). Cows (n = 173) that failed to express oestrus at 72 hr after PGF2α received 100 μg of GnRH and CS insemination concomitantly. Two weeks later, cows were penned with natural service sires (bull:cow ratio 1:25) for 45 days. Pregnancy was diagnosed 30 days after bull removal. Calves' gender was determined at birth. For cows that expressed oestrus, PR/AI did not differ (p > .1) between SS (65.0%) and CS (66.7%) groups. The overall PR/AI differed (p < .05) between SS (65.0%) and CS (56.4%) groups. The natural service PR differed (p < .001) but breeding season PR (p > .05) did not differ between SS vs. CS groups. Bull:heifer gender ratio following AI was 88:12 and 52:48 for SS and CS groups, respectively, with an overall 66:34 ratio. Bull:heifer gender ratio for the two breeding seasons was 79:21 and 52:48 for SS and CS groups, respectively, with an overall 62:38 ratio. In conclusion, the fertility of SS insemination at 18 hr after onset of oestrus was 97% of CS insemination at 12 hr after onset of oestrus. Though breeding season pregnancy did not differ between SS and groups, preferred calf gender was 25 percentage points greater for SS over CS application. The gender accuracy was 88%.     

Cryopreservation of llama semen using a combination of permeable cryoprotectants

Semen cryopreservation is not available for massive use in South American Camelids (SACs) due to the lack of an efficient protocol and the low pregnancy rates obtained with artificial insemination (AI). The use of a single cryoprotectant (CP) is commonly used in SACs frozen semen. The objective of the study was to evaluate the combined cryoprotective capacity of two permeable CPs at different stages of the cryopreservation protocol in llama semen. Sixteen ejaculates from 4 llama males were analysed, and sperm quality was assayed in raw semen, at 5°C, after equilibration of samples with the CPs and when samples were thawed. The following CPs and combination were used: 6% glycerol (GL), 6% dimethylformamide (DMF) and the combination of both CPs: 3% GL and 3% DMF. A Kruskal–Wallis test and an experimental factorial design, considering one factor with four levels (raw semen, 6% GL, 6% DMF and GL/DMF), were used. Total sperm motility and live sperm with intact acrosomes remained unchanged after equilibration of samples (p > .05). A significant decrease in the percentage of functional membrane, motile and live sperm with intact acrosomes was observed when samples were thawed (GL, DMF and GL/DMF). Nevertheless, the cryopreservation protocols used preserved sperm DNA quality; thus, sperm chromatin condensation and DNA fragmentation were unaffected (p > .05) when GL, DMF and GL/DMF were used. To conclude, no superiority was found between the use of a single or a combination of permeable cryoprotectants to freeze llama semen.     

Comparison of pregnancy per AI of heifers inseminated with sex-sorted or conventional semen after oestrus detection or timed artificial insemination

The objective of the study was to compare the fertility after using sex-sorted or conventional semen either with oestrus detection (EST) or timed artificial insemination (TAI) in Holstein heifers. Holstein heifers were randomly assigned to one of the following treatments in a 2 × 2 factorial design. Heifers in the EST group were inseminated with sex-sorted (n = 114) or conventional semen (n = 100) after spontaneous or induced oestrus. Heifers in the TAI, subjected to the 5-day Cosynch+Progesterone protocol (GnRH+P4 insertion-5d-PGF_2α+P4 removal-1d-PGF_2α-2d-GnRH+TAI), were inseminated with sex-sorted (n = 113) or conventional semen (n = 88). Statistical analyses were performed using PROC GLIMMIX procedure of SAS 9.4 (SAS Institute Inc., Cary, NC). Overall P/AI was 60.7% for EST and 54.2% for TAI regardless of types of semen and 68.1% for conventional and 48.9% for sex-sorted semen regardless of insemination strategies. Fertility of heifers inseminated with either sex-sorted (53.5%; 44.2%) or conventional (69.0%; 67.0%) semen did not differ between EST and TAI respectively. Besides, the interaction between the semen type and the insemination strategy was not significant for P/AI. The embryonic loss was significantly greater with sex-sorted semen (17.1%) compared to conventional semen (1.6%). There was no sire effect with sex-sorted semen on P/AI (52.6% vs. 46.2%) and embryonic loss (16.4% vs. 18.0%). As expected, sex-sorted semen resulted in more female calves (89.8% vs. 51.6%) than conventional semen. Thus, sex-sorted semen can be used with 5-day Cosynch+Progesterone protocol to eliminate the inadequate oestrus detection and to increase female calves born in dairy heifers.     

浅谈猪精液保存在人工授精中的发展与展望

在世界上主要的猪肉生产国中,猪人工授精(AI)技术的应用率在90%以上。尽管猪冷冻精液技术在不断进步,但由于其效率不高,在猪场实际生产中并不多见使用。在猪人工授精技术中,液态精液是目前应用最广泛的精液,与传统的牛冷冻精液不同,其独特的精子微胶囊保护膜技术,可以对猪精子进行更加完善的液态保存,因此液态精液是目前猪人工授精使用的主要方式。文章综述了猪精液保存技术在人工授精中的应用,并展望了今后的发展方向,为人工授精技术助力恢复生猪生产提供参考。     

Current challenges in sheep artificial insemination: A particular insight

Ovine artificial insemination (OAI) is not commonly performed because of specific problems related to semen application techniques, leading to highly variable results. The ideal methodology (frozen‐thawed semen/vaginal route) is unfeasible under field conditions due to the cervix morphology of the ewe, which prevents the process of intrauterine insemination necessary to obtain acceptable results. Currently, OAI commercial programmes use superficial cervical insemination, CAI (vaginal), with chilled semen (15°C) and intrauterine insemination, LAI (laparoscopic), with frozen‐thawed semen. The ability to improve upon these contrasting techniques may be derived from examining certain poorly studied factors such as insemination time, productive state of females and alternatives of seminal preservation, some of which we reviewed in this work. This interim solution will remain in use until AI by the vaginal route with frozen‐thawed semen is developed, but it poses new challenges in optimizing the freezing of the sperm and adapting the cervical (CAI) and/or transcervical intrauterine AI (TCAI). In this review, we address the current problems and evaluate their methodological (mechanical) and chemical (dilation) alternatives. Currently, TCAI is a methodologically complex technique with poor fertility results, so further studies are needed to improve the logistics of this procedure and the results of its application.     

Oxytocin increases pregnancy rates after fixed time artificial insemination in Kazak ewes

It is probable that reduced pregnancy rates in ewes after fixed time artificial insemination (FTAI) is attributable, in part, to the reduced number of normal spermatozoa that colonize the oviduct. Administration of oxytocin stimulates both cervical dilation and uterine/oviductal contractility. The hypothesis that oxytocin can enhance sperm transport into the uteri and the oviducts, and thereby increase pregnancy rates, was tested in the present study. Oestrus was synchronized in 199 multiparous Kazak ewes using intravaginal flurogestone-impregnated sponge. The sponge was left in the vagina for 12 days followed with an injection of 330 IU of eCG at sponge removal. Each ewe was intracervically inseminated twice at 50 hr and 62 hr after the removal of sponges using an insemination catheter containing 0.25 ml of diluted semen. Semen was collected from seven Texel rams and all the ejaculates were pooled and diluted in ultra-high temperature-treated commercial skimmed milk without (Control group, 0.05 ml of saline per mL milk, n = 144) or with oxytocin supplement (Oxytocin group, 0.5 U of oxytocin per ml milk, n = 55). Pregnancy status was determined by transabdominal ultrasound examination 45 days after insemination. Lambing performance was recorded at delivery. Significant differences were observed between the Oxytocin group and the Control group in terms of the pregnancy rate and the fecundity rate (85.5% and 92.7% versus 68.8% and 72.9%, respectively). In conclusion, low dose oxytocin supplementation of semen extender significantly increased pregnancy and fecundity rates in oestrus-synchronized Kazak ewes after FTAI.     

Reproductive performance of immobilized cryopreserved bovine semen used for timed artificial insemination

The SpermVital^® technology comprises embedding of spermatozoa within an alginate gel to facilitate release of sperm cells over a prolonged period in utero after AI. The aim of this study was to examine whether the survival time of spermatozoa is extended when applying this immobilization technology in combination with cryopreservation. Sperm cell survival (acrosome and plasma membrane integrity) was studied in vitro for 48 hr at physiological temperature. One dose of SpermVital^® (SV) semen was compared with single doses of Biladyl^® (B) processed semen as well as double doses of B (B double). B double was obtained by adding a second B dose the following day, thereby mimicking double AI. Furthermore, reproductive performance applying single early timed AI (TAI) with SV following oestrus synchronization was studied in a field trial. Double insemination (TAI on two consecutive days) with B semen served as control. Number of acrosome‐intact live sperm cells decreased over time in vitro for all treatments (p < .05). There was no difference between SV sperm cell survival and B double after 24 hr (p > .05). However, after 48 hr, SV sperm cell survival was higher than B double (p < .05). Moreover, multivariate analysis showed that the outcome of single early TAI with SV was not significantly different from B double (p > .05). Likelihood of pregnancy and calving in the heifer group was higher than in the cow group (p < .05). These results imply that spermatozoa immobilized in alginate gel have prolonged survival.     

Antibiotic resistance in microorganisms isolated in a bull semen stud

Many microorganisms from various sources may be present in ejaculates of bulls. This study identified and isolated bacteria from bull sperm samples in a commercial stud and evaluated their resistance to antibiotics. The number of colony-forming units was determined in semen samples collected at distinct steps during freezing and thawing. The minimum inhibitory concentration and the minimum bactericidal concentration were determined for four antibiotics commonly used in commercial studs. A total of 135 microorganisms from 25 genera were isolated. After a sensitivity test, all evaluated microorganisms (n = 55) were resistant to penicillin and most of them were resistant to tylosin and lincomycin (n = 54). Resistance to all tested antibiotics was observed in 22% of all isolates, whereas only 3.9% of the isolates were inhibited by the tested antibiotics at the concentrations recommended by the international legislation. As the isolated microorganisms presented high resistance to frequently used antibiotics, sensitivity tests should be periodically conducted in commercial bull semen studs to prevent the use of contaminated semen in artificial insemination.     

Fertility in Dairy Cows After Artificial Insemination Using Sex‐Sorted Sperm or Conventional Semen

The aim of this study was to compare pregnancy per artificial insemination (P/AI) after timed AI with sex‐sorted sperm (SS) or conventional semen (CS) in lactating dairy cows. Cyclic cows (n = 302) were synchronized by Ovsynch and randomly assigned into two groups at the time of AI. Cows with a follicle size between 12 and 18 mm and clear vaginal discharge at the time of AI were inseminated with either frozen‐thawed SS (n = 148) or CS (n = 154) of the same bull. A shallow uterine insemination was performed into the uterine horn ipsilateral to the side of probable impending ovulation. Pregnancy per AI on Day 31 tended (p = 0.09) to be less for SS (31.8%) than CS (40.9%). Similarly, P/AI on Day 62 was less (p = 0.01) for cows inseminated with SS (25.7%) compared with CS (39.0%). The increased difference in fertility between treatments from Days 31 to 62 was caused by the greater (p = 0.02) pregnancy loss for cows receiving SS (19.2%) than CS (4.8%). Cow parity (p = 0.02) and season (p < 0.01) when AI was performed were additional factors affecting fertility. Primiparous cows had greater P/AI than multiparous cows both on Day 31 (41.7% vs 25.0% in SS and 53.0% vs 31.8% in CS groups) and on Day 62 (33.3% vs 20.5% in SS and 48.5% vs 31.8% in CS groups). During the hot season of the year, P/AI on Day 31 was reduced (p = 0.01) in the SS group (19.6%) when compared with the rates during the cool season (38.1%). In conclusion, sex‐sorted sperm produced lower fertility results compared to conventional semen even after using some selection criteria to select most fertile cows.     

Resynchronization of ovulation with new and reused intravaginal progesterone‐releasing devices without previous pregnancy diagnosis in Bos taurus indicus cows subjected to timed‐artificial insemination

The study aimed to evaluate pregnancy per artificial insemination (P/AI) of cows subjected to synchronization and resynchronization in ovulation protocols using intravaginal progesterone‐releasing insert (P4) before pregnancy diagnosis (PD) and the relationship of PR with the diameter of preovulatory follicles (ØPOF) before TAI. Cows (n = 378) were distributed into two groups: a resynchronization group with new devices (GRN; n = 185) and resynchronization group with used devices (GRU; n = 193). On Day 0, both groups received a new P4 and estradiol benzoate (EB). On D8, P4 removal + D‐cloprostenol + eCG + estradiol cypionate (EC) was done. On d10, TAI was conducted. On d32, cows were resynchronized and divided into two groups, GRN (n = 185) and GRU (n = 193). The GRN group received a new P4 + EB, and the GRU group received a used P4 + EB. On d40, the P4 was removed + PD. The non‐pregnant cows received D‐cloprostenol + eCG + EC. US was done again on d42 to determine ØPOF before the second TAI. The P/AI of the GRN and GRU groups after synchronization were 56.2% and 57.0% (p = 0.87), respectively, and those after resynchronization were 58.0% and 37.3% (p < 0.008), respectively. The P/AI of the GRN and GRU groups observed after TAI (synchronization + resynchronization) were 81.6% and 73.1%, respectively (p = 0.047). No difference (p = 0.067) in ØPOF between the pregnant and non‐pregnant cows in the GRN was found, whereas the GRU group showed a significant difference (p = 0.003). Resynchronization protocols optimized the P/AI in both groups. New intravaginal devices resulted in greater P/AI and P/AI accumulation in resynchronization as compared with the GRU; the ØPOF was related with P/AI.