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Fertilization of IVF/ICSI using sibling oocytes from couples with subfertile male or unexplained infertility.

The significance of the performance of conventional in vitro fertilization and intracytoplasmic sperm injection (IVF/ICSI) using sibling oocytes from couples with subfertile male or unexplained infertility was evaluated. A total of 410 sibling oocyte cumulus-corona complexes (OCCC) from 21 couples with subfertile male (group A) and 11 unexplained infertile couples (group B) were randomly divided, in order of retrieval, into two groups inseminated either by conventional IVF or by ICSI. The treatment outcomes and the influence of infertility factors on fertilization in each group were compared. The results showed that although the two pronuclear (2PN) fertilization rate per injected sibling oocytes was significantly higher after ICSI (group A: 68.2% +/- 28.8%; group B: 66.2% +/- 24.9%) than after conventional IVF (group A: 41.8% +/- 32.7%; group B: 40.1% +/- 22.1%), the other variables studied included: the fertilization rates of per allocated sibling oocytes IVF/ICSI, the fertilization rates of sibling oocytes IVF/ICSI after excluding failed IVF fertilization cycles, as well as the cleavage rates of normal fertilization were not statistically significant (P>0.05). Similarly, though the total fertilization failure rate in the IVF group (group A: 42.9%; group B: 36.4%) was significantly higher than in the ICSI group (group A: 4.8%; group B: 0), we did not cancel cycles due to the normal fertilization of sibling oocytes. Embryo transfer was possible in all 32 couples. There were 10 clinical pregnancies in the two groups. We also discovered a possible association between some semen parameters and sperm functions of group A, and women age and duration of infertility of group B and fertilization. It is suggested that adoption of the split IVF/ICSI technology in the above cases may help eliminate fertilization failures. This is also a useful method to investigate the effect of single factor on the employment of assisted reproductive technology.

Embryo Transfer↗

[[Determinants of the decline in the total marital fertility rate in the early 1970s]].

The author examines "the relationship between the total marital fertility rate and the cohort marital fertility pattern in Japan since the late 1960s. We calculated parity-duration-year-specific marital fertility rates and conducted comparisons among them cohort by cohort and period by period. In addition, we carried out a decomposition of the change in total marital fertility rates into the tempo and quantum components on two different assumptions about future completed fertility. By calculating the first-birth, second-birth and third-birth timing indices, we could specify the effect due to a change of birth timing for each parity on the variation of the total marital fertility rate." It is found that "given that the drop in the total fertility rate in the early 1970s is largely attributable to a decrease in the total marital fertility rate, most of the drop in the total fertility rate in this period is also attributable to the change in second-birth timing over marriage cohorts since the middle 1960s and the temporal disturbance in second-birth timing in marriage cohorts of the early 1970s." (SUMMARY IN ENG)

Asia↗

Fertilization alters the spatial distribution and the density of voltage-dependent sodium current in the egg of the ascidian Boltenia villosa.

The spatial distribution of voltage-dependent ionic currents was characterized in Boltenia villosa eggs before and after fertilization using two-microelectrode voltage clamp of paired animal-vegetal halves of eggs (merogones) made surgically. Major voltage-dependent conductances in the Boltenia egg are a transient inward Na current, a transient inward Ca current, and an inwardly rectifying K current. These currents were randomly distributed along the animal-vegetal axis in the unfertilized egg. When paired merogones (surgically prepared egg fragments) were made at the vegetal cap stage, 15-30 min after fertilization, Ca and K currents remained randomly distributed along the animal-vegetal axis. In contrast, the relative Na current density was found to be twofold lower in the vegetal vs the animal merogones made at the vegetal cap stage. By making pairs of merogones from unfertilized eggs and subsequently fertilizing one merogone of a pair, we showed that this change in current density ratio was due to a loss of absolute Na current density in the vegetal hemisphere shortly after fertilization. These results also show that this loss was intrinsic to the vegetal hemisphere, rather than being determined solely by the point of sperm entry. A second decrease in Na current was observed during the hour before first cleavage, 60-120 min after fertilization (M.L. Block and W.J. Moody, 1987, J. Physiol. 393, 619-634), both in fertilized eggs and in animal merogones fertilized after isolation. This second loss of Na current was not observed in vegetal merogones fertilized after isolation or in either animal or vegetal merogones made from fertilized eggs at the vegetal cap stage. Possible mechanisms for te rapid (complete by 40 min after fertilization) and the late (occurring from ca. 60 to 120 minutes after fertilization) Na current losses are discussed.

Action Potentials↗

Analysis of human spermatozoal fertilizing ability using zona-free ova.

An in vitro fertilization assay employing zona-free hamster eggs was used to analyze human spermatozoal fertilizing ability. Human spermatozoa were preincubated for 18 to 20 hours in Biggers, Whitten, and Whittingham's medium (1971) at a concentration of 1 X 10(7) sperm/ml prior to the addition of zona-free superovulated hamster eggs. Eggs were examined microscopically 2 hours later for evidence of swelling or decondensing sperm heads in the cytoplasm. A total of 6266 eggs were examined in assays for both suspected fertile and infertile donors; 50 eggs/sample were examined. The percentage fertilization was found to range from 14% to 100% in the suspected fertile group with an average of 56.3%. The sperm concentration in this fertile group ranged from 22 to 303 million/ml with an average of 114. The suspected infertile samples yielded fertilization rates of 10% or less and an average count of 50.6 million/ml. These data suggest that human spermatozoa fuse with the vitelline membrane of zona-free hamster eggs and decondense with varying efficiencies. The percentage of fertilization in this cross-species system did not show a significant correlation with sperm concentration or motility. However, suspected infertile samples always yielded 10% or less fertilization in this assay. This method may have potential value as a diagnostic tool in evaluating human spermatozoal fertilizing capacity which avoids the ethical and logistcal problems associated with fertilization of human eggs in vitro.

Animals↗

Cumulus contributions during bovine fertilization in vitro.

A mandatory step in performing micromanipulation techniques, studying sperm-oocyte interactions and evaluating morphological aspects of oocyte quality is the removal of cumulus cells from oocytes or zygotes at various stages. In cattle, cumulus removal shortly before fertilization in vitro strongly decreases sperm penetration rates. This study was conducted to evaluate the function of the cumulus oophorus during bovine fertilization in vitro. The importance of cumulus secretions during IVF was investigated by inseminating cumulus-denuded oocytes (CDOs) in fertilization medium supplemented with individual cumulus secretions, such as progesterone or hyaluronic acid. None of these substances increased the fertilization rate of CDOs. However, fertilizing CDOs in cumulus-conditioned medium or on a cumulus monolayer partially restored the reduction in fertilization rate (P<0.05). The fertilization rate of CDOs inseminated on a cumulus monolayer further increased when physical contact between the gametes and the monolayer was prevented by fertilizing them inside a culture plate insert placed on the monolayer (P<0.05). Finally, the importance of reactive oxygen species (ROS) generation and O(2) concentration during IVF was studied. Luminol-dependent chemiluminescence revealed a higher ROS load in conditioned medium of cumulus-enclosed oocytes (CEOs) than in that of CDOs after sperm-oocyte co-incubation (P<0.05). Furthermore, lowering the external O(2) concentration from 20 to 5% decreased the fertilization rate of both CEOs and CDOs, but had a higher impact on CEOs (P<0.05). In conclusion, this study provides evidence that the cumulus oophorus benefits the fertilizing ability of penetrating spermatozoa by creating a complex microenvironment of both cumulus secretions and metabolic products around the oocyte. Gap junctional communication between the oocyte and corona cells as well as sperm trapping by the cumulus oophorus seem to be essential factors in supporting fertilization.

Animals↗

The importance of porcine sperm parameters on fertility in vivo.

It would be desirable to use semen parameters to predict the in vivo fertilizing capacity of a particular ejaculate. In animal production, an ejaculate is divided into multiple doses for artificial insemination (AI); therefore, it would be economically beneficial to know the functional quality (i.e., fertility) of the semen before it is inseminated. To identify a predictive assay of the fertilizing capacity of a porcine ejaculate, we performed 4 rapid assays of sperm quality (motility, viability, physiological status as assessed by chlortetracycline fluorescence, and ATP content) on samples from 9 ejaculates, before and after a thermal stress test (42.5 degrees C, 45 min). These parameters were subsequently correlated with in vivo fertility resulting from AI with 2 sperm doses, 3 x 10(9) or 0.3 x 10(9) motile cells in 70 mL (optimal or suboptimal sperm number per insemination, respectively) from these same ejaculates. No parameter was correlated to the fertility rates obtained after inseminating with the optimal semen doses, either before or after the thermal stress test (P > 0.05). However, with respect to the animals inseminated with the suboptimal semen dose, sperm motility (the percentage of motile spermatozoa as assessed visually by microscopy) prior to thermal stress was well-correlated to fertility rates (r = 0.783, P = 0.01). The percentage of spermatozoa displaying the chlortetracycline Pattern AR (acrosome reaction) was also statistically related to fertility (r = 0.05, P = 0.04), but the biological importance of this relationship is questionable given the small variation among ejaculates (range: 0 to 2%). No other sperm parameter was significantly related to fertility rates in this group (P > 0.05). These data, therefore, indicate that sperm motility is a useful indicator of sperm fertilizing capacity in vivo. Moreover, to identify a predictor of semen fertility it is critical that the number of spermatozoa used during insemination is sufficiently low to detect differences in sperm fertilizing efficiency.

Adenosine Triphosphate↗

The cost of a successful delivery with in vitro fertilization.

BACKGROUND: The use of in vitro fertilization has engendered considerable debate about who should have the procedure, whether health insurance should cover the cost, and if so, to what extent. We investigated the cost of a successful delivery with in vitro fertilization. METHODS: We calculated the cost per successful delivery with in vitro fertilization (defined as at least one live birth) for a general population of couples undergoing in vitro fertilization and for two subgroups: couples with a diagnosis of tubal disease (who have a better chance of success), and couples in which the woman is over the age of 40 years and the man has a low sperm count (who have a lower chance of success). Information on charges per cycle of in vitro fertilization was obtained from six facilities across the country; delivery rates with this procedure were estimated from the literature. RESULTS: On average, the cost incurred per successful delivery with in vitro fertilization increases from $66,667 for the first cycle of in vitro fertilization to $114,286 by the sixth cycle. The cost increases because with each cycle in which fertilization fails, the probability that a subsequent effort will be successful declines. Sensitivity analyses indicated that the cost per delivery ranges from $44,000 to $211,940. For couples with a better chance of successful in vitro fertilization (i.e., those with a diagnosis of tubal disease), it costs $50,000 per delivery for the first cycle and $72,727 for the sixth. For couples in which the woman is older and there is a diagnosis of male-factor infertility, the cost rises from $160,000 for the first cycle to $800,000 for the sixth. CONCLUSIONS: The debate about insurance coverage for in vitro fertilization must take into account ethical judgments and social values. But analyses of costs and cost effectiveness help elucidate the economic implications of using in vitro fertilization and thus inform the policy discussion.

Adult↗

Placental villitis of unclear etiology during ovum donor in vitro fertilization pregnancy.

OBJECTIVE: Preliminary observations by a single pathologist at our institution revealed a 75% incidence of villitis of unexplained etiology in ovum donor in vitro fertilization pregnancies. Because the incidence of villitis of unexplained etiology in the general population is approximately 10%, we conducted a controlled study to compare the incidence of villitis of unexplained etiology in ovum donor in vitro fertilization pregnancies to that in in vitro fertilization pregnancies that do not use donated ova. STUDY DESIGN: Placental specimens of ovum donor in vitro fertilization pregnancies were matched randomly with pregnancies that resulted from both fresh and frozen/thawed native oocyte in vitro fertilization from March 5, 1995, to October 10, 2001, and examined in a blinded fashion by a single pathologist (D. J. R.) for villitis of unexplained cause. The incidence of villitis of unexplained etiology was analyzed in 27 patients who underwent ovum donor in vitro fertilization versus 37 patients who underwent native oocyte in vitro fertilization. RESULTS: Villitis of unexplained cause occurred in 22.2% of ovum donor in vitro fertilization pregnancies, 10.8% of native oocyte in vitro fertilization pregnancies (fresh and frozen/thawed combined), and 14.3% of frozen/thawed cycles (P=.21). CONCLUSION: Although the incidence was not statistically different than in in vitro fertilization that used native maternal oocytes, there was a 2-fold increase in villitis of unexplained cause in the ovum donor in vitro fertilization placentas, which suggests that immune-related disorders may be increased in ovum donor pregnancies.

Adult↗

The recent rise in Palestinian fertility: permanent or transient?

The purpose of this paper is to describe trends in fertility and fertility change in the West Bank and Gaza Strip during the years 1968-1992, with a particular focus on the recent rise in fertility. The paper is based primarily on vital registration data. The findings show that the fertility transition has been well underway in the West Bank since 1985, with a lull during the Intifada period. No indication of a fertility transition in the Gaza Strip was found. An examination of the age pattern of fertility reveals that Palestinian fertility has been increasingly shifting toward younger ages, with a particular concentration in the age group 15-24 during the Intifada period. A decomposition analysis of the fertility rates shows that the recent rise in fertility is essentially due to changes in marital structure, but also to increased fertility within marriage among younger women. No evidence of increased fertility among older women is found. Some implications of the findings are discussed.

Arabs↗

Phenotypic traits as reliable indicators of fertility in male broiler breeders.

Genetic selection procedures applied to improve broiler performance may negatively impact the subsequent reproductive efficiency of breeders, particularly in males. Identification of traits that reliably indicate individual male fertility would facilitate selection for reproduction. We hypothesized that physical traits, such as comb area, relative testicular weight, and testicular weight asymmetry, may correlate with fertility in two male-selected primary broiler breeder strains (A and B). Thirty males per strain, individually housed with an average of 10 females, were evaluated at five age periods within the 30-to-50-wk breeding cycle. Flock fertility by candling eggs at Day 19 of incubation and sample fertility by visual assessment of the germinal disc were determined. Sperm penetration (SP) through the perivitelline layer was assessed. Comb area was evaluated by image analysis at 40 and 50 wk, and relative testicular weight was measured at 50 wk. Strain A sample and flock fertility (P < 0.001) and SP values (P < 0.0001) were significantly lower than Strain B. Both strains had a significant decline of fertility and SP with age (P < 0.0001). Strain A comb area correlated with sample fertility (P < 0.05), flock fertility (P < 0.05), and relative testicular weight (P < 0.01). Conversely, Strain B relative testicular weight correlated with sample fertility (P < 0.0001) and flock fertility (P < 0.001). Significant correlations were not found between testicular weight asymmetry and other reproductive traits. Results suggest comb area may be a reliable indicator of male fertility in Strain A.

Age Factors↗

The inheritance of modifiers conferring self-fertility in the partially self-incompatible perennial, Campanula rapunculoides L. (Campanulaceae).

The role of partial self-incompatibility in plant breeding system evolution has received little attention. Here, we examine the genetic basis of modifiers conferring self-fertility in the creeping bellflower, Campanula rapunculoides L. (Campanulaceae), a partially self-incompatible herb. A survey of 35 individuals from two natural populations indicates that 45% of them are strongly self-incompatible, 40% intermediately self-incompatible, and 15% weakly self-incompatible and that some plants show a strong breakdown in self-incompatibility over floral age. We generated 101 F1 families by random crossing among 31 parental plants and estimated the heritability of self-fertility in day 1 and day 4 female-phase flowers, the genetic correlation between day 1 and day 4 self-fertility, and the coefficient of additive genetic variance of self-fertility. We use linear regression and data from additional crosses to examine whether there are significant maternal effects in the expression of self-fertility. We use Fain's test to determine if a major gene influences self-fertility and, finding no evidence, use data from additional crosses on an F2 generation to estimate the mean number and dominance of genes conferring self-fertility. These analyses indicate that the heritability (h2) of self-fertility is 0.24 in day 1 female-phase flowers and 0.44 in day 4 flowers, self-fertility is primarily additive but shows some recessive effects, and self-fertility is estimated to be controlled by four genetic factors. In addition, we have evidence that there may be maternal effects for self-fertility, especially for weakly self-incompatible plants. The significance of these results in the context of mating system evolution is discussed.

Biological Evolution↗

Timing of phosphorus fertilizer application within an irrigation cycle for perennial pasture.

Irrigated pastures are significant contributors of phosphorus (P) to inland watercourses, with much of the P coming from applied fertilizer. It was hypothesized that the timing of P fertilizer application relative to irrigation regulates P concentrations in runoff and infiltrating water. To test this hypothesis, a two-by-two factorial experiment was conducted on twelve 8- x 30-m border-irrigated bays growing perennial pasture. Phosphorus fertilizer in the form of single superphosphate (44 kg P ha(-1)) was surface-broadcast onto the bays when the nominal change in soil water deficit reached 0 or 50 mm (U.S. Class A pan evaporation minus rainfall). Following fertilizer application, the bays were again irrigated when the nominal soil water deficit between fertilizing and the subsequent irrigation reached either 0 or 50 mm. The volume of water applied, runoff volume, and changes in soil water content were recorded for the three irrigations following fertilizer application. Total phosphorus (TP) and filtrable reactive phosphorus (FRP, <0.45 microm) concentrations in runoff and at depths of 0.1, 0.3, and 0.6 m in the soil were also measured. Soil water content at fertilizer application had less effect on P concentrations in runoff and soil water than the additional time between fertilizing and irrigating. By allowing a deficit of 50 mm between fertilizer application and irrigation, the average concentration of P in runoff and moving below a soil depth of 0.1 m was approximately halved. To maximize fertilizer use efficiency and minimize environmental effects, a delay should occur between applying P fertilizer and irrigating perennial pasture.

Agriculture↗

The importance of seminal plasma on the fertility of subsequent artificial inseminations in swine.

Yorkshire x Landrace sows and gilts were used in a 3x2 factorial arrangement of treatments to determine the effect of uterine inflammation induced by either killed spermatozoa (KS) or bacterial lipopolysaccharide (LPS) on the fertility of a subsequent, optimally timed AI. Estrus was detected with a mature boar twice daily. Twelve hours after the first detection of estrus, females received intrauterine infusions of an inflammatory stimulus consisting of a 100-mL dose of extender containing 3x10(9) KS (n = 40), 20 microg of LPS (n = 40; positive control) or extender alone (n = 40; negative control). An insemination was performed 12 to 18 h later with 3x10(9) motile spermatozoa (i.e., fertile AI) suspended in either 100 mL of seminal plasma (SP; n = 60) or extender replenished with of estrogens (5 microg of estradiol-17beta, 4.5 microg of estrone sulfate, and 2 microg of estrone; n= 60). Transcutaneous ultrasound was performed at the time of fertile AI and again 24 h later to detect the presence or absence of preovulatory follicles. A fertile AI performed within 24 h before ovulation was considered optimal. Conception (CR) and farrowing rates (FR) were greater in females that received a fertile AI diluted with SP compared with extender (P<.01), and there was a significant (P<.05) treatment x fertile AI dilution medium interaction for both CR and FR. Females that received a fertile AI 12 h after infusion of extender had similar CR and FR regardless of fertile AI dilution medium. After inducing an inflammatory response with either KS or LPS, CR and FR were higher in females that received a fertile AI diluted with SP compared with fertile AI dilution with extender (P<.05). The effects of treatment and AI dilution media and their interactions were not significant for litter size in females that farrowed. These results show that the fertility of a subsequent AI can be impaired when semen is deposited into an inflamed environment created by an earlier AI, and this impairment was offset by inclusion of SP in the subsequent insemination.

Animals↗

Duration of the seasonal anestrus in sheep selected for fertility in a fall-lambing system.

Crossbred ewes (1/2 Dorset, 1/4 Rambouillet, and 1/4 Finnsheep) from a flock being selected for spring fertility, defined as ability to lamb following ram exposure in May and June in Virginia (37 degrees N latitude), were used to study the duration of the seasonal anestrus. In the first 3 yr of the study (1992, 1993, and 1995), mature ewes were divergently selected based on EBV for fertility, and the duration of anestrus was measured by continuously exposing the ewes to vasectomized rams equipped with marking harnesses from mid-January until approximately August 1. Only ewes that had lambed in the previous fall were used to ensure that ewes were in a comparable physiological state, and the same rams remained with the ewes in each year to avoid induction of estrus by introduction of novel rams. The duration of anestrus in high-fertility ewes (n = 26; mean fertility EBV of 12.6%) was 28.4 d, which was significantly less than the 70.2 d of anestrus observed for low-fertility ewes (n = 15; mean fertility EBV of .3%). Five high-fertility ewes did not exhibit a period of anestrus. The regression of number of days of anestrus on fertility EBV was -2.15 +/- .72 d/%. In yr 4 (1997), 11 high-fertility and two low-fertility ewes were evaluated. None of these ewes exhibited a clear seasonal anestrus; six unequivocally cycled continuously. Between January 23 and July 31, the mean duration of anestrus for these ewes was only approximately 11 d. The duration of anestrus for high-fertility ewes seems to be the shortest reported for temperate sheep breeds.

Anestrus↗

Effect of ammonium sulfate fertilization on bahiagrass quality and copper metabolism in grazing beef cattle.

To assess the impact of S fertilization on bahiagrass (Paspalum notatum) quality and Cu metabolism in cattle, two studies were conducted during the summer grazing season (1999 and 2000). Pasture replicates (16.2 ha; n = 2/treatment) received the same fertilizer treatment in each growing season, consisting of 1) 67 kg N/ha from ammonium sulfate (AS), 2) 67 kg N/ha from ammonium nitrate (AN), and 3) control (no fertilizer; C). Forage sampling was conducted at 28-d intervals following fertilization by the collection of whole plants (four samples/pasture) in randomly distributed 1-m2 grazing exclusion cages and analyzed for CP, in vitro organic matter digestibility, S, P, Ca, K, Mg, Na, Fe, Al, Mn, Cu, and Zn. To determine the effect of fertilizer treatment on liver trace mineral concentrations in grazing cattle, random liver tissue samples were collected (n = 12; four/treatment) at the start and end of the study period in 2000. Ammonium sulfate fertilization increased (P < 0.001) forage S concentration in both years. Plant tissue N concentrations were increased by N fertilization, regardless of source, in 2000, but not in 1999. Cows grazing AS pastures had lower (P < 0.05) liver Cu concentrations at the end of the study period in 2000 compared to AN and C. In Exp. 2, 37 Cu-deficient heifers grazing AS fertilized pastures were obtained from the same location and allocated to one of two treatments, consisting of supplements providing 123 mg/d of either inorganic (Cu sulfate; n = 12) or organic (Availa-Cu; n = 15) Cu. Treatments were delivered for 83 d. Liver Cu increased over time in all heifers regardless of treatment; however, heifers supplemented with Availa-Cu tended (P = 0.09) to have higher mean liver Cu concentrations than those receiving Cu sulfate. The results of these studies indicate that AS fertilization of bahiagrass increases forage S concentrations. When provided free-choice access to a complete salt-based trace mineral supplement, cows grazing AS-fertilized pastures had lower liver Cu concentrations than cows grazing pastures fertilized with AN; upon removal from high-S pastures, cattle were able to respond to Cu supplementation.

Ammonium Sulfate↗

Influence of herd environment on health and fertility and their relationship with milk production.

High levels of milk production in dairy cattle can have negative side effects on health and fertility traits. This paper explores the relationships among milk yield, health, and fertility traits both across and within herd environments on a national scale. A total of 456,574 lactations from 3904 herds recorded from 1995 to 1999 in The Netherlands were analyzed. Herd environment was defined by 41 variables derived from production records and the annual national agricultural survey. Principal components analysis reduced this set to 4 components: intensity, defined as average production per cow, average fertility, farm size, and relative performance indicating whether herds had good (poor) health and fertility despite a high (low) production. Both fertility and health were better for some traits in high-intensity herds and for other traits in low-intensity herds. In high-intensity herds, somatic cell count (SCC) levels were lower, drops in production occurred more often, and first service took place earlier but with lower success. High fertility occurred more often in herds located on sandy soils and in those that had lower SCC levels, had fewer drops in production and higher cow survival. On large farms, drops in production were less frequent and fertility was somewhat better. The within-herd analysis showed that the relationship of milk yield with health and fertility was stronger in herds with high production, fertility, or both. In herds with poor relative performance, there was no difference in production levels between animals with good health or fertility and those with poor health or fertility.

Animals↗

Relationship of bull fertility to sperm nuclear shape.

The relationship between sperm nuclear shape and bull fertility was determined. Two groups of bulls, 3 per group, were selected. Bulls differed in fertility based on lifetime nonreturn rates. Digital images of propidium iodide-stained sperm from each bull were collected and shape-evaluated by Fourier harmonic amplitudes 0 to 5. A discriminant function (P < .05) was constructed based on harmonic amplitudes and the 2 fertility groups. When individual sperm were classified as being of high or lower fertility, the percentage of each bull's sperm placed in the high-fertility group had a linear relationship (r = .89, P < .05) with fertility. To construct a plot of mean sperm shapes, a novel technique to automatically orient and identify the anterior tip of the sperm head was developed. The mean nuclear shape of high-fertility sperm was more elongated and tapered than those of lower fertility. A discriminant function (P < .05) was also constructed that separated the 6 bulls into 2 groups based only on the harmonic amplitudes or sperm nuclear shape. The bulls were correctly classified into the 2 fertility groups. A comparison of sperm chromatin structure analysis (SCSA) and harmonic amplitudes found that overall size variance, anterior roundness, and posterior taperedness of sperm nuclei were related to chromatin stability (P < .05). Some of the differences observed in sperm nuclear shape between the high- and lower-fertility bulls may be explained by varying levels of chromatin stability. However, sperm nuclear shape appears to contain additional information from chromatin stability alone. In this particular study, with 6 bulls, all with good chromatin quality, sperm nuclear shape was a better predictor of bull fertility.

Animals↗

Influence of seminal additives and packaging systems on fertility of frozen bovine spermatozoa.

The following recommendations and conclusions are based upon results of fertility and laboratory studies, and general trends from field investigations. Fertility results due to the addition of enzymes have been variable and contradictory. Flushing of ampules with dry, gaseous nitrogen prior to filling has become a routine practice in processing semen to be frozen. For control of Vibrio fetus and Leptospira pomona, 2,000 micrograms of streptomycin and 1,000 u polymyxin B sulfate should be added per milliliter of raw semen immediately after collection. The extender for initial dilution should contain the same concentration of antibiotics used for raw semen plus 500 u penicillin. The glycerol portion of the extender should contain 500 u penicillin per milliliter. The effect of addition of sugars on fertility has been highly variable. The primary beneficial effect is probably due to their cryoprotective properties. A myriad of concoctions have been added to bovine semen and the results have been highly variable with respect to both motility and fertility. Results of subsequent experiments have rarely proven that addition of exotic compounds or mixtures has been of value. Higher mean fertility was obtained with semen in straws in 14 of 21 comparisons with ampules. The differences in favor of straws ranged from 1.1 to 18.9; while the range in favor of ampules was .1 to 4.4 percentage points. Fertility obtained with pellets has ranged from minus 12.8 to plus 11.9 percentage points in nonreturn rate (NR), compared to the corresponding NR with semen in ampules. Fertility of semen in ampules was higher in five of eight studies. Fertility of pelleted semen has ranged from minus 9.5 to plus 6.0 percentage points compared with straws. Fertility was higher for semen in pellets in only one of five investigations. Pellets should not be used until the potential for pathogenic contamination and exchange of spermatozoa among pellets is eliminated. There is a potential for higher fertility with semen in straws as compared to other packaging systems, but the issue of liquid nitrogen (LN) entry and possible contamination of semen should be further investigated. In general, fertility obtained with semen frozen in the .25 ml straw has been equal to or higher than semen in larger packages. However, they cannot be unequivocally recommended due to other considerations. From laboratory studies, it appears that greater spermatozoan survival is obtained when semen frozen in straws is thawed in water at 35 C or above.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗