Use of a semen extender containing antibiotic to improve the fertility of a stallion with seminal vesiculitis due to Pseudomonas aeruginosa

A breeding trial was conducted to determine if a semen extender containing polymixin-B sulfate would improve the fertility of a stallion with seminal vesiculitis due to Pseudomonas aeruginosa . Twenty-three mares were bred to the stallion by one of three methods: artificial insemination with raw semen (Group 1, n = 10), artificial insemination with semen mixed 1:1 with a nonfat dry skim milk/glucose extender containing 1000 units/ml polymixin-B sulfate (Group 2, n = 9), or natural service immediately following infusion of the uterus with 100 ml of the same extender (Group 3, n = 4). Artificial breedings contained a minimum insemination dose of 500 x 10(6) progressively motile spermatozoa. All mares were bred every other day while in estrus. Pregnancy status was determined by transrectal ultrasound examination 15 d after the last breeding. First-cycle pregnancy rate for Group 2 mares (78%) was greater (P < 0.01) than for Group 1 mares (10%). There was a tendency (P = 0.10) for the pregnancy rate of Group 3 mares (50%) to be greater than Group 1 mares. The use of a semen extender containing polymixin-B sulfate improved the fertility of this stallion.     

Follicular dynamics and concentrations of steroids and gonadotropins in lactating cows and nulliparous heifers

Differences in follicular development and circulating hormone concentrations, between lactating cows and nulliparous heifers, that may relate to differences in fertility between the groups, were examined. Multiparous, cyclic, lactating Holstein cows (n=19) and cyclic heifers (n=20) were examined in the winter, during one estrous cycle. The examinations included ultrasound monitoring and daily blood sampling. Distributions of two-wave and three-wave cycles were similar in the two groups: 79 and 21% in cows, 70 and 30% in heifers, respectively. Cycle lengths were shorter by 2.6 days in heifers than in cows, and in two-wave than in three-wave cycles. The ovulatory follicle was smaller in heifers than in cows (13.0+/-0.3 mm versus 16.5+/-0.05 mm). The greater numbers of large follicles in cows than in heifers corresponded well to the higher concentrations of FSH in cows. The duration of dominance of the ovulatory follicle tended to be longer in cows than in heifers. Estradiol concentrations around estrus and the preovulatory LH surge were higher in heifers than in cows (20 versus 9 ng/ml). Progesterone concentrations were higher in heifers than in cows from Day 3 to Day 16 of the cycle. Circulating progesterone did not differ between two-wave and three-wave cycles. The results revealed differences in ovarian follicular dynamics, and in plasma concentrations of steroids and gonadotropins; these may account for the differences in fertility between nulliparous heifers and multiparous lactating cows.     

Side of gestation in dairy heifers affects subsequent sperm transport and pregnancy rates after deep insemination into one uterine horn

Effects of side of previous gestation on sperm transport and pregnancy rates after deep cornual insemination were evaluated in 1686 Friesian cows in their first lactational period. Only single ovulating animals were used. At insemination, semen was deposited deep into the uterine horn ipsilateral or contralateral to the preovulatory follicle. A total of 876 cows (52%) ovulated in the ovary ipsilateral to the postgravid horn, and 810 cows ovulated in the contralateral ovary. Semen was deposited into the previously nongravid uterine horn of 832 cows, and into the gravid horn of 854 cows. The pregnancy rate was higher (P < 0.00001) for semen deposition into the previously nongravid horn (46.6%) than for semen deposition into the gravid horn (35.7%). For inseminations ipsilateral to the side of impending ovulation, pregnancy rates were higher (P = 0.0004) when ovulations occurred on the opposite side to the postgravid horn than on the same side. Pregnancy rates were higher (P = 0.002) for contralateral inseminations when ovulations occurred on the same side to the postgravid horn than on the opposite side; they were higher (P = 0.0001) for total ipsilateral than for total contralateral inseminations. There was no difference between ipsilateral and contralateral inseminations (P = 0.64) when ovulation occurred ipsilateral to the postgravid horn, but pregnancy rates were higher (P < 0.00001) when ipsilateral insemination was carried out into the nonpostgravid horn. Results indicate that the side of gestation in dairy heifers affects subsequent pregnancy rates after deep insemination into one uterine horn, possibly by affecting sperm transport.     

Effect of the metabolic environment at key stages of follicle development in cattle: focus on steroid biosynthesis

Cellular mechanisms that contribute to low estradiol concentrations produced by the preovulatory ovarian follicle in cattle with a compromised metabolic status are largely unknown. To gain insight into the main metabolic mechanisms affecting preovulatory follicle function, two different animal models were used. Experiment 1 compared Holstein-Friesian nonlactating heifers (n = 17) and lactating cows (n = 16) at three stages of preovulatory follicle development: 1) newly selected dominant follicle in the luteal phase (Selection), 2) follicular phase before the LH surge (Differentiation), and 3) preovulatory phase after the LH surge (Luteinization). Experiment 2 compared newly selected dominant follicles in the luteal phase in beef heifers fed a diet of 1.2 times maintenance (M, n = 8) or 0.4 M (n = 11). Lactating cows and 0.4 M beef heifers had higher concentrations of β-hydroxybutyrate, and lower concentrations of glucose, insulin, and IGF-I compared with dairy heifers and 1.2 M beef heifers, respectively. In lactating cows this altered metabolic environment was associated with reduced dominant follicle estradiol and progesterone synthesis during Differentiation and Luteinization, respectively, and in 0.4 M beef heifers with reduced dominant follicle estradiol synthesis. Using a combination of RNA sequencing, Ingenuity Pathway Analysis, and qRT-PCR validation, we identified several important molecular markers involved in steroid biosynthesis, such as the expression of steroidogenic acute regulatory protein (STAR) within developing dominant follicles, to be downregulated by the catabolic state. Based on this, we propose that the adverse metabolic environment caused by lactation or nutritional restriction decreases preovulatory follicle function mainly by affecting cholesterol transport into the mitochondria to initiate steroidogenesis.     

Feeling the ovaries prior to insemination. Clinical implications for improving the fertility of the dairy cow

During the periovulatory period in dairy cattle, the largest ovarian follicle can be felt by palpation per rectum as a firm/soft follicle (young preovulatory follicle), a very soft follicle separating it from the remainder of the ovary (mature preovulatory follicle), or an evacuated follicle (follicle associated with ovulation). Because any one of these three follicle types may be present at the time of artificial insemination, the objective of this study was to identify possible differences between the effects of a firm/soft, very soft, or evacuated ovarian follicle on fertility. Out of a study sample of 2365 inseminations, very soft, firm/soft, and evacuated follicles were recorded in 1689 (71%), 593 (25%), and 83 (3.5%) inseminations, respectively. Logistic regression analysis indicated no significant effects of largest follicle type, vaginal discharge, season, days in milk, parity, synchronized or natural estrus, and semen-providing bull on the pregnancy rate. The only variable included in the final logistic regression model was the interaction season-follicle type. This interaction determined that the likelihood of pregnancy decreased significantly by factors of 0.86 or 0.82 in cows with a firm/soft follicle inseminated during the cool or warm period, respectively, and by a factor of 0.09 in cows with evacuated follicles inseminated during the warm period, using as reference cows with a very soft follicle inseminated during the cool period (yielding the highest pregnancy rate). As an overall conclusion, the state of the periovulatory follicle at insemination was clearly related to fertility and masked the effects of factors commonly affecting fertility such as parity, days in milk at AI and inseminating bull. More importantly they suggest that by including ovarian follicle checks in artificial insemination routines, the success of this procedure could be improved.     

Effect of insemination with doses of 2 or 15 million frozen-thawed spermatozoa and semen deposition site on pregnancy rate in dairy cows

The effects of low-dose artificial insemination (AI) on pregnancy rates have seldom been studied in lactating dairy cows. We evaluated the pregnancy results after AI with doses of 2 and 15 million frozen-thawed spermatozoa and the effect of semen deposition in lactating dairy cows. A total of 284 first inseminations with 2 million spermatozoa and 312 first inseminations with 15 million spermatozoa were performed on 480 dairy farms. Low-dose inseminations (2 million spermatozoa) under field conditions in commercial dairy herds, without estrus synchronization, generally resulted in significantly reduced pregnancy rates compared with normal doses (15 million spermatozoa). The bull x technician effect on fertility was statistically significant. This finding indicates that there is a high variability in fertility among bulls using 2 million spermatozoa per dose. The semen deposition site did not influence pregnancy rates. It is concluded that a dose of 2 million frozen-thawed spermatozoa is probably too low for most bulls to achieve acceptable pregnancy rates in dairy cows.     

Effects of different artificial insemination techniques and sperm doses on fertility of normal mares and mares with abnormal reproductive history

The effects of different artificial insemination (AI) techniques and sperm doses on pregnancy rates of normal Hanoverian breed mares and mares with a history of barrenness or pregnancy failure using fresh or frozen-thawed sperm were investigated. The material included 187 normal mares (148 foaling and 39 young maiden mares) and 85 problem mares with abnormal reproductive history. Mares were randomly allotted into groups with respect to AI technique (routine AI into the uterine body, transrectally controlled deep intracornual AI ipsilateral to the preovulatory follicle, or hysteroscopic AI onto the uterotubal junction ipsilateral to the preovulatory follicle), storage method of semen (fresh, frozen-thawed), AI volume (0.5, 2, 12 ml), and sperm dose (50 x 10(6) or 300 x 10(6) progressively motile sperm (pms) for fresh semen and 100 or 800 x 10(6) frozen-thawed sperm with >35% post-thaw motility). The mares were inseminated once per cycle, 24 h after hCG administration when fresh semen was used, or 30 h for frozen-thawed semen. Differences in pregnancy rates between treatment groups were analyzed by Chi-squared test, and for most relevant factors (insemination technique, mare, semen, and stallion) expectation values and confidence intervals were calculated using multivariate logistic models. Neither insemination technique, volume, sperm dose, nor mare or stallion had significant effects (P > 0.05) on fertility. Type of semen, breeding mares during foal heat, and an interaction between insemination technique, semen parameters, and mares did have significant effects (P < 0.05). In problem mares, frozen semen AI yielded significantly lower pregnancy rates than fresh semen AI (16/43, 37.2% versus 25/42, 59.5%), but this was not the case in normal mares. In normal mares, hysteroscopic AI with fresh semen gave significantly (P < 0.05) better pregnancy rates than uterine body AI (27/38, 71% versus 18/38, 47.3%), whereas in problem mares this resulted in significantly lower pregnancy rates than uterine body AI (5/15, 33.3% versus 16/19, 84.2%). Our results demonstrate that for problem mares, conventional insemination into the uterine body appears to be superior to hysteroscopic insemination and in normal mares, the highest pregnancy rates can be expected by hysteroscopic insemination.     

Effect of the inseminate and the site of insemination on the uterus and pregnancy rates of mares

In this review, effects of the composition of the inseminate on uterine response and pregnancy rates in mares are discussed. The inseminate can differ for volume, sperm concentration, total sperm numbers, presence, absence, or proportion of seminal plasma, and extender composition. Semen can be used as fresh, cooled, or frozen. The site of semen deposition also plays a role; semen is deposited either into the uterine body (standard artificial insemination (AI)) or into the tip of the uterine horn ipsilateral to the preovulatory follicle (deep AI) using the hysterocopical or transrectally guided techniques. In addition to pregnancy rates, some uterine responses to the inseminate are considered including myometrial contractions, transport and elimination of sperm, and uterine inflammation, which is reflected as numbers of polymorphonuclear leukocytes, enzyme levels, and presence of intrauterine fluid. Reproductively normal and abnormal mares are compared.     

The effect of semen extender,seminal plasma and raw semen on uterine and ovarian blood flow in mares

Transrectal color Doppler sonography was used to evaluate the effect of intrauterine infusion of skim milk semen extender, seminal plasma and raw semen on the endometrium and blood flow in the uterine and ovarian arteries in mares. Six Trotter mares (mean age: 12 years) were examined during estrus in three cycles. Each mare received an intrauterine infusion of 20 ml of skim milk semen extender, seminal plasma or raw semen during estrus in one of three cycles. Blood flow measurements in both uterine and ovarian arteries and the determination of intrauterine fluid via sonography were performed before each infusion and 1, 3, 6, 12, and 24 h after infusion. Forty-eight hours later, the intrauterine infusion and measurements were repeated using the same time intervals. Changes in blood flow were detected using transrectal color Doppler sonography and were evaluated using the mean time-averaged maximum velocity (TAMV) of the blood flow. Cytological and bacteriological examination of uterine swabs performed 48 h after the second infusion revealed less inflammation and bacterial growth in mares infused with skim milk semen extender than in those infused with seminal plasma or raw semen. There was an increase in intrauterine fluid as early as 1 h after infusion of any of the substances. The infusion of skim milk semen extender had no effect on uterine blood flow. Within 1 h after infusion of seminal plasma or raw semen, there was an increase in the TAMV values of both uterine arteries (P<0.05). In contrast, ovarian blood flow increased only in the artery ipsilateral to the preovulatory follicle and only after the infusion of raw semen (P<0.05). In conclusion, the changes in uterine perfusion observed after intrauterine infusion may be associated with endometrial inflammation and vasodilatory components in the seminal plasma, whereas the changes seen in ovarian blood flow are possibly attributable to the interaction between sperm and oviduct.     

Antibodies to egg yolk in blood serum of rabbits and cattle and cervical mucus of cattle inseminated artificially.

Serum titers to egg yolk were induced in 6 rabbits by intravaginal deposition of an egg-yolk citrate extender used for artificial insemination of cattle. There was no effect of the low serum titers to egg yolk on fertility of the inseminated rabbits. Titers to egg-yolk semen extender were found in 3% of 59 cows of normal fertility compared to 29% of 14 repeat breeder cows of low fertility, all previously inseminated with semen diluted with egg yolk-citrate extender. Four of 6 cervical mucus samples (67%) from the repeat breeder cows had high titers to egg yolk, but only one also had a positive titer in blood serum.     

Factors affecting the yield of viable embryos by superovulated Holstein-Friesian cows

GnRH treatment (250 ug) 48 h after prostaglandin F(2alpha) in 40 superovulated cows induced a release of LH (increment > 5 ng/ml) in only 13 of the older cows. Eleven of these cows did not yield viable embryos. Thirty-two of 75 cows had preovulatory surge levels of LH 48 h after prostaglandin treatment. Plasma progesterone concentrations were determined in 140 cows at the time that superovulation was initiated. Eighty-four of these donors were superovulated with 40 mg of FSH and 56 donors with 48 mg of FSH. There was no relationship (P > 0.05) between the concentration of progesterone at the start of superovulation with either ovulation rate determined by palpation per rectum or the number of viable embryos per flush. These parameters were also unaffected (P > 0.05) by age of the donor or the dose of FSH. In another group of donors, treatment with 40 mg FSH was compared over a 3-d (n = 28) and a 4-d (n = 18) interval. The donors treated with FSH over a 3-d period had similar ovulation rates but yielded less viable embryos (1.5 v 5.8, P < 0.05). The fertility rate of 33 cows, inseminated 60 and 72 h after prostaglandin, was comparable to the fertility rate of 18 cows inseminated at 60, 72 and 84 h after prostaglandin treatment.     

Effect of L-carnitine administration on the seminal characteristics of oligoasthenospermic stallions

The effect of orally administered l-carnitine on the quality of semen obtained from stallions with different semen qualities was investigated. Four stallions with proven fertility (high motility group, HM) and with normal seminal characteristics (>50% progressive motility and > 80 x 10(6) spermatozoa/ml), and four questionable breeders (low motility group, LM) with <50% of sperm progressive motility and < 80 x 10(6) spermatozoa/ml, received p.o. 20 g of l-carnitine for 60 days. Blood and semen samples were collected before treatment (T0) and after 30 (T1) and 60 days (T2). Semen evaluation were performed on five consecutive daily ejaculates (n = 120 ejaculates) and conventional semen analysis was carried out on each ejaculate, both at collection and after refrigeration for 24, 48, and 72 h. Furthermore l-carnitine, acetylcarnitine, pyruvate, and lactate concentrations, and carnitine acetyltransferase activity (CAT) were determined both in raw semen and seminal plasma. There were an increase in progressive motile spermatozoa only in the LM group (26.8 +/- 12.9, 39.1 +/- 15.5, and 48.8 +/- 8.6 for T0, T1, and T2, respectively). Free seminal plasma carnitine concentration was higher in the LM group compared to the HM one. Both pyruvate and lactate were higher in the LM group. Raw semen and seminal plasma carnitine and acetylcarnitine levels correlate positively with both sperm concentration and progressive motility; moreover, acetylcarnitine content was positively correlated with total motile morphologically normal spermatozoa. In conclusion, oral administration of l-carnitine to stallions with questionable seminal characteristics may improve spermatozoa kinetics and morphological characteristics; whereas, it seem to be ineffective in normospermic animals.     

Low semen dose intracornual insemination of cows at fixed time after PGF2alpha treatment or at spontaneous estrus

The numbers of spermatozoa per insemination and the site of semen deposition in the uterine horn appear to interact to influence pregnancy rate. In two experiments, the effect of a single low dose (2 x 10(6) spermatozoa) intracornual insemination (LD-ICI) on bovine pregnancy rate was compared with that of intracornual (SD-ICI) and conventional (SD-AI) inseminations of 40 x 10(6) spermatozoa. In Experiment 1, 157 cows were treated twice with PGF(2)alpha at a 14-day interval and inseminated at a fixed time (80-82 h) after the second PGF(2)alpha injection using LD-ICI (n=44), SD-ICI (n=61) or SD-AI (n=52). In LD-ICI and SD-ICI groups, semen was deposited in the horn ipsilateral to the ovulatory follicle close to the utero-tubal junction (LD-ICI-UTJ, n=33 and SD-ICI-UTJ, n=41) or in the middle part of the horn (LD-ICI-MH, n=11 and SD-ICI-MH, n=20). Pregnancy rates after LD-ICI-UTJ, LD-ICI-MH, SD-ICI-UTJ and SD-ICI-MH were 27%, 27%, 39% and 35%, respectively (P>0.05). The total pregnancy rate after LD-ICI (27%) did not differ (P>0.05) from that after SD-ICI (37%) or SD-AI (34%). In Experiment 2 (field trial), 362 cows were allotted, at spontaneous estrus, to LD-ICI-UTJ (n=86), LD-ICI-MH (n=97) or SD-AI (n=179). Pregnancy rates after LD-ICI and SD-AI were 47% and 45%, respectively (P>0.05). After LD-ICI-UTJ, the pregnancy rate (54%) did not differ significantly (P>0.05) to that obtained after LD-ICI-MH (41%) and after SD-AI (45%). The results of the study show that the single intracornual insemination of cows with 2 x 10(6) spermatozoa at fixed time, 80-82 h after the second PGF(2)alpha injection or at spontaneous estrus resulted in similar pregnancy percentage as intracornual and conventional inseminations with 40 x 10(6) spermatozoa per semen dose. With intracornual insemination using low or standard dose of spermatozoa, the pregnancy rates were not significantly affected by the exact site of semen deposition in the uterine horn, near the utero-tubal junction or in the middle part.     

Effect of length of progesterone exposure during ovulatory wave development on pregnancy rate

The objective was to determine the effects of the duration of progesterone exposure during the ovulatory wave on fertility (pregnancy rate) in beef cattle. We tested the hypothesis that short-progesterone exposure during the growing and early-static phase of the ovulatory follicle (analogous to the ovulatory wave of 3-wave cycles) is associated with higher fertility than a longer duration of exposure (analogous to the ovulatory wave of 2-wave cycles). Three to 5 days after ovulation, beef heifers (n = 172) and suckled beef cows (n = 193) were given an intravaginal progesterone-releasing device (CIDR) and 2.5 mg estradiol - 17β +50 mg progesterone im to induce a new follicular wave. Cattle were allocated to short- or long-progesterone exposure groups (for 3 and 6 d after wave emergence, respectively) after which prostaglandin F_2α was administered and CIDR were removed. Forty-eight hours later, all cattle were given 12.5 mg pLH and artificially inseminated (AI) with frozen-thawed semen. The diameter of the two largest follicles and the corpus luteum were measured by transrectal ultrasonography at CIDR removal, insemination, and 36 h after insemination. Pregnancy diagnosis was done ultrasonically 38 and 65 d post-AI. There was no difference in pregnancy rates in short- vs long-progesterone exposure in heifers (53 vs 47%, P = 0.44) or cows (63 vs 58%, P = 0.51). However, the diameter of the ovulatory follicle at CIDR removal and AI was smaller in short- than in long-progesterone groups (P < 0.02), and larger in cows than in heifers (P < 0.006). In conclusion, short-progesterone exposure during the growing and early-static phase of the ovulatory follicle (similar to 3-wave cycles) was not associated with higher fertility than a longer progesterone exposure (similar to 2-wave cycles).     

Fertilization rates in superovulating cows after deposition of semen on the infundibulum, near the uterotubal junction or after insemination with high numbers of sperm

Three experiments were conducted with 105 superovulating Holstein dairy cows in attempts to improve the fertilization rate. Cows were superovulated with follicle-stimulating hormone (FSH) and time of estrus was regulated with prostaglandin F(2)alpha (PGF(2)alpha). Semen was deposited on each infundibulum through a laparoscope inserted through the flank (Experiment 1) or near the uterotubal junctions through flexible tubing passed through the cervix and uterine horns (Experiment 2). In the third experiment, high numbers of sperm in fresh semen were deposited in the uterus. Cows were necropsied and ova were recovered and examined about 3.5 d after the beginning of estrus. Deposition of 0.5 ml of frozen-thawed semen on each infundibulum (Experiment 1) reduced both ovum recovery and fertilization. In ten cows inseminated on the infundibulum, ova representing 43% of ovulation points were recovered and 9% of these recovered ova were fertilized. In ten control cows, ova representing 80% of ovulation points were recovered and 62% of them were fertilized. In a 2 x 2 experiment with 36 superovulating cows (Experiment 2), 1 ml of diluted fresh or frozen semen was deposited either near the uterotubal junction or in the uterine body. The overall fertilization rate was 61%, with no significant effect of site of semen deposition or type of semen used. In Experiment 3, 2 or 3 ml of neat semen (average of 4.4 billion sperm) was deposited in the uterus of 12 cows; 183 of 197 intact ova (93%) were fertilized. In 56 control cows inseminated with 0.5 to 1.5 ml of frozen diluted semen (average of 70 million sperm), 502 of 947 intact ova were fertilized (53%, P<0.001). Insemination with high numbers of fresh sperm overcame problems of sperm loss or sperm transport and improved the fertilization rate.     

Successful pregnancies with directional freezing of large volume buck semen

Artificial insemination with frozen-thawed buck semen shows variable results which depend on many factors related to semen quality and the cryopreservation processing. We conducted experiments based on a new freezing method, directional freezing, of large volumes (8 ml). In the first experiment semen from three Saanen bucks, ages 1-2-years-old and genetically selected for milk improvement, was frozen individually. Two to three-years-old Saanen females (n = 164) were synchronized with controlled internal drug release (CIDR), pregnant mare serum gonadotrophin (PMSG) and prostaglandin. Double cervical inseminations were performed with frozen-thawed semen and fresh semen as control. In the second experiment we used pooled, washed frozen semen to examine the effect of washed seminal plasma. The motility after washing was 80-90% and after thawing was 55-65% for all bucks. The sperm concentration increased with the collections and the advance into the breeding season from 1.9 x 10(9) to 4.4 x 10(9) cell/ml average. Two inseminations were carried out at 8h intervals. The first insemination was performed at 32 h after CIDR withdrawal with fresh and frozen-thawed semen. Pregnancy rates were assessed by ultrasonography conducted 40 and 90 days post-insemination (from three bucks). Results were 58, 67, 50% with fresh semen, and for frozen semen were 33, 37 and 53%; these results were significantly different in one of the three bucks (P < 0.005). In the second experiment with pooled, washed semen the pregnancy rate was 41.6%, which compared with the average results of the frozen semen in the first experiment 38.9% no significant difference was found. We conclude that freezing buck semen in large volumes (8 ml) is possible. Cryobanking of buck semen will facilitate a genetic breeding program in goats and preservation of biodiversity. Washed semen did not improve the fertility of the semen when Andromed bull extender is used.     

Site of semen deposition in cattle: a review

The breeding of cattle using conventional artificial insemination methods involves the deposition of semen in the uterine body. However, it has been recently proposed by several authors that the site of semen deposition be changed to the uterine horns. This suggestion is based on 2 facts: the acceptance that the major preovulatory sperm reservoir may be the uterotubal junction rather than the cervical canal, and the lack of accuracy by inseminators in depositing semen. In over 50% of cases, inseminators were not sufficiently trained to deposit semen into the uterine body, so that intracervical insemination was often performed resulting in reduced fertility. The advantage of deep uterine insemination, whether bicornual or unicornual, is that it favors the deposition of semen nearer to the uterotubal junction and thus reduces the incidence of cervical deposition. This review updates the literature on the ideal site of semen deposition, including cervical, uterine body, cornual and intraperitoneal. Also analyzed are the effects of right vs. left side activity of the female reproductive tract on the optimum site of semen deposition as it affects fertilization. Finally, the question of whether the clinical training of inseminators should be reevaluated is discussed.     

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%).     

Fixed time deep intracornual insemination of heifers at synchronized estrus

The aim of the study was to determine the efficiency of single fixed time deep intracornual insemination using 2 x 10(6) spermatozoa compared with single standard dose deep intracornual insemination and single and dual standard dose (40 x 10(6)) uterine body (conventional) insemination in heifers at synchronized estrus. Estrus was synchronized in 275 virgin heifers by administration of two doses of PGF(2)alpha 14 days apart. Deep intracornual inseminations with low (ICI-LD1, n=102) and standard (ICI-SD1, n=56) dose of semen and the single standard dose conventional inseminations (AI-SD1, n=66) were performed 80-82 h after the second PGF(2)alpha treatment. Ultrasonography was used to identify the first dominant (presumed ovulatory) follicle, and semen was deposited either close to the utero-tubal junction (n=69 in ICI-LD1 and n=23 in ICI-SD1) or in the middle part of the uterine horn (n=28 in ICI-LD1 and n=28 in ICI-SD1) ipsilateral to the ovary bearing the first dominant follicle. The dual standard dose conventional inseminations were performed 72 and 96 h after the second PGF(2)alpha treatment (AI-SD2, n=51). The pregnancy rate in the ICI-LD1 group (68.0%) did not differ significantly (P>0.05) from the ICI-SD1 group (56.9%) or the AI-SD2 group (65.9%) and was significantly higher (P<0.05) than in the AI-SD1 group (54.2%). The site of intacornual deposition of semen, near the utero-tubal junction or in the middle of the horn, had no effect on the pregnancy rate. The pregnancy rate in all the groups was not affected by the intensity of expression of estrous signs.     

A new antibiotic combination for frozen bovine semen, 3. Evaluation of fertility

Field fertility (nonreturn rate) studies were performed independently by three artificial insemination organizations to evaluate bovine semen processed for freezing using the antibiotics gentamicin, tylosin and Linco-Spectin at concentrations of 500 ug, 100 ug, and 300/600 ug, respectively, per milliliter of neat semen and per milliliter of nonglycerol portion of the extender. The antibiotic combination including penicillin, dihydrostreptomycin, polymyxin B sulfate, with/without Linco-Spectin (500 units/ml, 2000 ug/ml, 1000 units/ml and 300/600 ug/ml, respectively) was used as the control treatment. Results indicated no significant effect on seminal quality as measured by field fertility under the conditions of these experiments using heated whole milk or egg yolk-sodium citrate seminal extenders. Use of the new antibiotic combination has been adopted by Certified Semen Services.