Number Of Live Offspring Research Articles

Prenatal to peripubertal exposure to Di(2-ethylhexyl) phthalate induced endometrial atrophy and fibrosis in female mice

Di(2-ethylhexy) phthalate (DEHP) is a widely used plasticizer that is ubiquitously found in the environment. Using a mouse model, we investigated the impact of early life DEHP exposure ranging from the prenatal to peripubertal developmental period of the female reproductive system. Pregnant female mice were allocated to three groups as follows: control, 100 mg/kg/day, and 500 mg/kg/day DEHP treatment. DEHP exposure was introduced through feeding during pregnancy (3 weeks) and lactation (3 weeks). After weaning, the offspring were also exposed to DEHP through feeding for another 2 weeks. Observations were conducted on female offspring at 10 and 24 weeks. The number of live offspring per dam was significantly lower in the high-DEHP-exposed group (500 mg/kg/day) compared to the control group (7.67 ± 1.24 vs. 14.17 ± 0.31; p < 0.05) despite no difference in pregnancy rates across the groups. Low-DEHP exposure (100 mg/kg/day) resulted to a decreased body weight (36.07 ± 3.78 vs. 50.11 ± 2.11 g; p < 0.05) and decreased left uterine length (10.60 ± 1.34 vs. 14.77 ± 0.82 mm; p < 0.05) in 24-week- old female mice. As early as 10 weeks, endometrial atrophy and fibrosis were observed, and endometrial cystic hyperplasia was noted in female mice at 24 weeks. Our study is the first to demonstrate that female mice exposed to DEHP in the early life developed endometrial fibrosis in the female offspring. Further studies on the consequences of these observations in fecundity and other reproductive functions are warranted.

P-003 Sperm retrieved by microdissection testicular sperm extraction (MD-TESE) is associated with altered sex ratio of offspring after intracytoplasmic sperm injection in men with non-obstructive azoospermia

Abstract Study question Does the sperm retrieved surgically by MD-TESE of men with Non-Obstructive Azoospermia affect the live birth sex ratio? Summary answer Our data indicates that sperm retrieved in men with non-obstructive azoospermia alters the sex ratio in favor of female offspring. What is known already Non-Obstructive Azoospermia is the most severe form of male infertility and is the most common cause of Azoospermia. Among the different surgical techniques, MD-TESE gives the highest rate of sperm retrieval and when combined with intracytoplasmic sperm injection (ICSI), live births had been achieved with variable success, Sex ratio of offspring can vary substantially due to several variables, however no studies have investigated the resultant birth sex ratio in this setting. Study design, size, duration Retrospective Cohort study. Data were collected between May 2009 and December 2019. A total of 100 consecutive men with non-obstructive Azoospermia underwent MD-TESE for sperm retrieval. 46 couples underwent ICSI and live birth rate, cumulative live birth rate and sex ratio of offspring analysed. Participants/materials, setting, methods One-hundred men underwent MD-TESE out of which 46 proceeded with ICSI. Demographic data including male and female age, testicular volumes and serum hormone values are given as mean ± SD with a range (minimum and maximum). Fertility outcomes including sperm retrieval, fertilization rate, number of embryos transferred, and live birth rates and cumulative live birth rates were recorded. Chi-square test was performed to compare the proportions. Main results and the role of chance Sperm retrieval was successful in 58%(58/100). Testicular volume and hormonal parameters did not vary among patients with positive or negative sperm retrieval. Histology gave best correlation with sperm retrieval. Hypospermatogenesis yielded the highest sperm retrieval 93%(26/29), followed by Maturation arrest 78%(7/9), then Hyalinosis 46% (6/13) and lastly Sertoli cell only 38%(19/50). 46 couples underwent ICSI cycles where the mean age of patients and their wives were 36.4 ± 3.7y and 33.0 ± 4.3y. Fertilization rate and mean number of transferred embryos were 51.4 % and 1.7. Live birth rate and Cumulative Live Birth rate per Embryo transfer were 60.5% (26/43) and 74.4% (32/43) and per started ICSI cycle were 56.5% (26/46) and 69.6% (32/46), with a twin rate of 15.3%(4/26). Mean gestational length and mean body weight at birth were 39.0 ± 1.4w and 3228.5 ± 5.5 g. Number of live offspring were 36 (Female: Male = 26:10) giving rise to sex ratio of 2.6 to1 in favor of female offspring (P &amp;lt; 0.05). There were no neonatal death, and one baby had phenylketonuria. Limitations, reasons for caution This is the first study to test the hypothesis of sex ratio variation related to the source of sperm; the strength of our study is that all procedures were performed by the same surgeon, so there are no operator-dependent differences. This finding needs to be confirmed in larger cohort studies. Wider implications of the findings It has been demonstrated that different stressors may alter the sex of the offspring. While in the present study the stressor is the parental hypogonadism associated with non-obstructive azoospermia, We hypothesise that testicular environment may direct spermatogenesis in favor of X carrying sperm around the time of sperm retrieval. Trial registration number N/A

INNOVATIVE ULTRA-RAPID VITRIFICATION METHOD FOR FIVE OR FEWER TESTICULAR SPERMATOZOA FROM NON-OBSTRUCTIVE AZOOSPERMIC MEN AFTER MICROSURGICAL TESTICULAR SPERM EXTRACTION (MICRO-TESE)

Clinical outcomes of ICSI when using only five or fewer testicular spermatozoa after cryopreservation have been unsuccessful and are considered to be inferior to those using testicular fresh spermatozoa from Micro-TESE. A possible cause of these poor results has been the lack of a successful freezing technique. In these cases, repeated Micro-TESE and simultaneous oocyte pick up has been the only available treatment. We developed an innovative ultra-rapid vitrification method for five or fewer testicular spermatozoa from non-obstructive azoospermic men after Micro-TESE. This study included 113 non-obstructive azoospermic men using Micro-TESE, it was conducted retrospectively at St. Mother Clinic between 2011 and 2018. Each motile spermatozoon was carefully aspirated tail first into the pipette, put into a 2-μl microdroplet media of the vitrification medium near the tip of the Cryotop (Kitazato Corporation, Tokyo, Japan) submerged in liquid nitrogen vapor for 2 min and then immediately plunged into liquid nitrogen. The vitrified spermatozoa were warmed by dipping them into a droplet media. Successfully recovered motile sperm were selected and used for ICSI. Number of patients, transfer cycles and collected sperms were 113, 192 and 560. Mean age of patients and their wives were 32.0±3.7y and 28.4±5.8y. Clinical pregnancy rate, miscarriage rate, live birth rate and number of live offspring were 24.0% (46/192), 19.6% (9/46), 19.3% (37/192) and 37 (Male: Female = 17: 20). Sperm recovery rate and survival rate were 90.3% (506/560) and 70.4% (356/506). Fertilization rate and mean number of transferred embryos were 51.6% (99/192) and 1.73 (1-2). Mean gestational weeks and mean body weight at birth were 39.23±5.27w and 2852.31±314.28g. No congenital anomalies were observed in any of the babies. This method for cryopreserving five or fewer spermatozoa from non-obstructive azoospermic men showed a recovery rate above 90% and a survival rate of about 70% which is the highest data ever been reported. The clinical pregnancy rate was 24.0% and 37 healthy babies were born.

P–023 Innovative ultra-rapid vitrification method for five or fewer testicular spermatozoa from non-obstructive azoospermic men after microsurgical testicular sperm extraction (Micro-TESE)

Abstract Study question What technique can be used to successfully cryopreserve five or fewer testicular spermatozoa from non-obstructive azoospermic men? Summary answer This method for cryopreserving five or fewer spermatozoa from non-obstructive azoospermic men showed a recovery rate above 90% and a survival rate of about 70%. What is known already Clinical outcomes of ICSI when using only five or fewer testicular spermatozoa after cryopreservation have been unsuccessful and are considered to be inferior to those using testicular fresh spermatozoa from Micro-TESE. A possible cause of these poor results has been the lack of a successful freezing technique. In these cases, repeated Micro-TESE and simultaneous oocyte pick up has been the only available treatment. Study design, size, duration Evaluation of the efficiency of cryopreservation by modified permeable cryoprotectant-free vitrification method (HTF supplemented with 0.1M sucrose and 10% SPS) for five or fewer testicular spermatozoa from 113 non-obstructive azoospermic men using Micro-TESE was conducted retrospectively at St. Mother Clinic between 2011 and 2018. Participants/materials, setting, methods This study included 113 non-obstructive azoospermic men. Each motile spermatozoon was carefully aspirated tail first into the pipette, put into a 2-μl microdroplet media of the vitrification medium near the tip of the Cryotop (Kitazato Corporation, Tokyo, Japan) submerged in liquid nitrogen vapor for 2 min and then immediately plunged in liquid nitrogen. The vitrified spermatozoa were warmed by dipping them into a droplet media. Successfully recovered motile sperm were selected and used for ICSI. Main results and the role of chance Number of patients, transfer cycles and collected sperms were 113, 192 and 560. Mean age of patients and their wives were 32.0±3.7y and 28.4±5.8y. Clinical pregnancy rate, miscarriage rate, live birth rate and number of live offspring were 24.0% (46/192), 19.6% (9/46), 19.3% (37/192) and 37 (Male: Female = 17: 20). Sperm recovery rate and survival rate were 90.3% (506/560) and 70.4% (356/506). Fertilization rate and mean number of transferred embryos were 51.6% (99/192) and 1.73 (1–2). Mean gestational weeks and mean body weight at birth were 39.23±5.27w and 2852.31±314.28g. No congenital anomalies were observed in any of the babies. Limitations, reasons for caution The maximum number of spermatozoa to which this method can be applied successfully is about 10. When the number of aspirated spermatozoa is over 10, some of them change direction and reach the mineral oil, and once this happens, they cannot be expelled out of the pipette. Wider implications of the findings: This technique is very useful for the cryopreservation of very small numbers of testicular spermatozoa (fewer than 10) in order to avoid or reduce Micro-TESE interventions. Trial registration number N/A

MON-035 Does Androgen Exposure Result in Germline Transmission of PCOS-Like Phenotypes and Can It Be Reversed?

Daughters of women with polycystic ovary syndrome (PCOS) are more likely to be diagnosed with PCOS, including reproductive and metabolic dysfunctions. Our recent research has demonstrated that dihydrotestosterone (DHT) exposure during late pregnancy results in transgenerational transmission of PCOS susceptibility to female offspring. But it remains unclear whether the transmission of the PCOS-like phenotypes is induced by in utero environment or via germ cell reprogramming, and whether treatment by exercise or androgen receptor blocker, flutamide, can prevent disease transmission. To model PCOS condition, donor mice were implanted with a continuous releasing DHT or vehicle pellet at 4 weeks of age. A subset of DHT exposed F0 donors had either free access to running wheels or were implanted with a slow-releasing flutamide pellet. Mice were exposed with or without treatment for either 6 weeks before IVF or 10 weeks prior to phenotypic testing. Here we present the phenotype of the F0 donors and the result of IVF to generate first (F1) and second (F2) generation offspring. Donors weigh more already after 2 weeks of DHT exposure and had more fat mass with larger adipocyte size, impaired glucose tolerance, and heavier kidney after 10 weeks of androgenization, which was reversed by both flutamide and exercise intervention. Moreover, DHT exposure increased circulating androgens and donors were completely acyclic. Simultaneous treatment with flutamide reversed the elevated androstenedione, testosterone, and restored estrus cyclicity, indicating that androgen receptor blocker can reverse hyperandrogenemia and reproductive dysfunction, whereas exercise failed to improve these phenotypes. After 6 weeks of exposure or treatment, donor oocytes were superovulated for IVF. Fewer oocytes per donor were found in androgenized + flutamide lineage, but no significant difference was observed in oocyte to two-cell embryo conversion rate after fertilization among all groups. Although the number of live offspring at weaning was similar among all groups, a trend of more F1 male than female offspring was found in both androgenized and androgenized + exercise lineage. Similar results were obtained in the F1 females when generating F2 offspring by IVF, which androgenized + flutamide lineage showed fewer oocytes per donor upon superovulation and more F2 male than female offspring was obtained in androgenized lineage at weaning. We here show that the androgenized donors develop clear PCOS-like phenotypes and give rise to more male than female F1 and F2 offspring. While blocking androgen receptor reverses both metabolic and reproductive disturbance in the donor, it also shows a negative impact on the donor and F1 female oocyte maturation process, although number of offspring via IVF is not affected. Excercise, however, only reverses the metabolic phenotypes in the F0 donor mice with no impact on IVF outcome.

Cryptorchidism and sex ratio are associated in dogs and pigs1

The objective of this study was to investigate whether the occurrence of cryptorchidism in a litter is associated with the sex ratio in this litter. The analysis included for dog 1,339 litters of 4 different breeds, of which 12.8% had 1 and 3.1% had 2 cryptorchids, and for pig 119,920 litters of 14 different populations, of which 2.2% had 1 and 0.2% had 2 cryptorchids. The data were analyzed using the GLIMMIX procedure of SAS. In dog, the presence of cryptorchids in a litter does not affect litter size or the stillborn rate. In pig, litter size increases significantly with an increasing number of cryptorchids in a litter. The stillborn rate in pig is larger in litters with cryptorchids than in litters without cryptorchids. To characterize the imbalance of the sexes in a litter, the sex ratio, defined as the number of live males divided by the number of live offspring, and the sex difference, defined as the number of live males minus the number of live females, were used as dependent variables in the analyses. In both dog and pig, the presence of 1 or 2 cryptorchids in a litter clearly goes together with a shift in both the sex ratio and the sex difference in a litter, in favor of male offspring. Our results in dog and pig suggest a common underlying mechanism affecting both cryptorchidism and sex ratio, which may hold true in other mammals as well.

149 LAPAROSCOPIC EMBRYO TRANSFER IN PIGS

In an effort to optimize the number of live offspring from cloned and transgenic pig embryos, embryos are surgically transferred into the isthmus region of the oviduct soon after micromanipulation. Surgical embryo transfer in pigs is successful but still an invasive process. Laparoscopic embryo transfer (Besenfelder et al. 1997 Theriogenology 47, 1051–1060) is much less invasive than surgery and is more adaptable to a variety of commercial embryo transfer conditions. Our goal was to develop the use the laparoscope as an alternative method of embryo transfer for micromanipulated embryos. Naturally cycling maternal white line donor and recipient females were used for laparoscopic embryo transfer. Donors were selected to be in estrus 0 to 24 h before recipients. Two- to four-cell embryos were surgically recovered via mid-ventral laparotomy and immediately prepared for laparoscopic transfer into the oviduct through the infundibulum or through puncture of the oviduct into the ampulla. Sixty-nine embryos (range, 13–20) were transferred into the oviduct via the infundibulum of four recipients. Two recipients farrowed (50%), one recipient spontaneously aborted on Day 27, and one returned to estrus on Day 25 of the estrous cycle. Twenty-one pigs were born (10.5 pigs/farrowed sow) resulting in 30% (21/69) of transferred embryos becoming live offspring. Ninety-seven embryos (range, 10-21) were transferred into the ampulla of the oviduct of six recipients. Four recipients farrowed (67%) and 2 returned to estrus on Day 20 and Day 28. Thirty-nine pigs were born (9.5/sow farrowed) resulting in 39% (38/97) of transferred embryos becoming live offspring. Observations indicate that it is much easier to find the oviduct and deliver embryos via puncture into the ampulla than find and insert the transfer catheter into the infundibulum. Using the efficiencies generated from these data, 1.1 more pigs can be expected per transfer by transferring embryos to the oviduct via puncture than into the oviduct via the infundibulum. These data demonstrate the effective use of the laparoscope for oviductal embryo transfer. Further work is needed to determine if the laparoscope improves the production of live offspring compared to surgical embryo transfer.

Inbreeding levels and prey abundance interact to determine fecundity in natural populations of two species of wolf spider

Long-term effective population size is expected, and has been shown, to correlate positively with various measures of population fitness. Here we examine the interacting effects of population size (as a surrogate for genetic factors) and prey consumption rates (as a surrogate for environmental quality) on fecundity in two sympatric species of wolf spider, Rabidosa punctulata and Rabidosa rabida. Population size was estimated in each of seven genetically isolated populations in each of 3 years using mark-recapture methods. Fecundity was estimated as the mean number of live offspring produced by ∼15 females sampled from each population of each species each year for 3 years. Prey consumption rates were estimated by sampling ∼300 spiders per population per year and assaying the proportion of spiders with prey. Larger populations have higher fecundity and more genetic diversity than smaller populations. Variation among populations in fecundity for a given year could be attributed most strongly to differences in population size, with variation in prey consumption rates and the interaction between population size and prey consumption playing smaller but still important roles. During the most stressful environmental conditions, the smallest populations of both species experienced disproportionately low-fecundity rates, more than doubling the estimated number of lethal equivalents during those years. The evidence presented in this paper for inbreeding-environment interactions at the population level and further evidence for a log-linear relationship between population size and fitness have important implications for conservation.

DETERMINATION OF PESTICIDES AND CHRONIC TEST WITH Daphnia magna FOR RAINWATER SAMPLES

Pesticides contained in rainwater were investigated in June and July 2001 in Yokohama. Although there were few farms and rice paddies near the sampling point, insecticides (fenitrothion and dichlorvos) and fungicides (chlorothalonil and futhalide) were detected by GC-MS and GC-FPD analysis. The maximum concentrations of dichlorvos and fenitrothion were 0.22 μg/L and 0.15 μg/L, respectively. Chronic tests with Daphnia magna were then conducted on two rainwater samples. Test solutions for these chronic tests were prepared by solid-phase extraction. In the case of the rainwater sampled on 25 July, the survival rate of D. magna was 100% after 21 days and no differences were found between the control and sample with regard to the average total number of live offspring produced per parent animal. However, a similar test with rainwater sampled on 14–15 June showed a marked toxic effect, since all the test animals were dead within 2 days. It is concluded that rainwater containing pesticides is an important ecological factor for freshwater plankton.

Ancillary Tests of Bull Semen Quality

Seminal quality examinations readily identify animals with low fertility, but rarely can these tests discriminate among males with moderate to high fertility. The new, automated semen analysis systems tend to be helpful in providing more reliable information. Such systems, especially the CASA systems that assess sperm morphology and motility, are innovative and certainly attractive. They may not, however, be cost effective. Similarly, although the combination of fluorescent staining and flow cytometry offers a very rapid and precise means of assessing the functional status of sperm organelles, it tends to be relatively expensive as a clinical tool. Such automated sperm quality assessments will likely become routine analyses at the larger semen-processing organizations. However, the true success of a breeding program can only be assessed by the number of live offspring. Thus, determination of true fertility, because of the complexity of the processes that unite the gametes, will continue to require more than a detailed examination of seminal cytology, no matter how sophisticated our methods become.

Reproductive Allocation in the Terrestrial Isopods Porcellio scaber and Oniscus asellus in a Metal-Polluted Environment

1. Terrestrial isopods (woodlice) exposed to toxic concentrations of metals can suffer early mortality and reduced body size, suggesting that the cost of metal detoxification causes trade-offs in resource allocation. This study investigates whether woodlice from populations that persist in sites heavily polluted with metals have a level of reproductive success different from that of woodlice from less polluted sites. 2. Two species of woodlice, Porcellio scaber Latreille and Oniscus asellus L., were collected from eight sites at different distances from an industrial smelting works at Avonmouth, England. The sites represented a continuum of concentrations of Zn, Cd, Pb and Cu in the woodlice, from background concentrations to a grossly contaminated site close to the smelter. Gravid females were reared in the laboratory until the release of the brood. 3. For both species there was no significant difference among sites in the number of live offspring per adult mass, the degree of brood-pouch mortality in affected adults, or the reproductive allocation. 4. Significant differences were found among sites in the proportion of adults experiencing some degree of brood-pouch mortality, and in the mean dry mass of offspring per adult mass. However, these differences could not be attributed to contamination by metals. 5. Females of both species that survive to reproduce in the most polluted sites around the smelter show no evidence of detrimental effects on reproduction. However, high concentrations of Zn at these sites causes early mortality of woodlice and therefore reduces the size of the breeding population.

Reproductive and developmental toxicity study of a new antineoplastic agent, S-1 (IV)--Perinatal and postnatal study in rats by oral administration

S-1 is a newly developed antineoplastic agent consisting in a molar ratio of 1:0.4:1 mixture of tegafur (FT), 5-chloro-2, 4, dihydroxypyridine (CDHP) and potassium oxonate (Oxo) was administered orally to SD rats at doses of 0, 1, 4 and 7 mg/kg/day (as a dose of FT) during the perinatal and postnatal periods to examine its effect on dams and postnatal growth of the offspring. A group as the control was treated only with medium (0.5% hydroxypropyl methylcellulose) solution. The administration of 7 mg/kg/day to dams caused suppression in body weight gains and in food consumption during the treatment period. No adverse effects of S-1 on the length of gestation, gestation index, delivery and nursing ability were found. The administration of 4 and 7 mg/kg/day caused suppression in body weight gains in offspring of both sexes. Significant decrease in kidney weights were observed in females of the 4 mg/kg/day group and in both sexes of the 7 mg/kg/day group. No adverse effects of S-1 were found in number of live offspring at birth, sex ratio of live offspring, number of dead offspring at birth, birth index, viability index, weaning index, incidence of external anomalies, general conditions, postnatal development, reflex responses, motor coordination, emotional behavior, learning ability, skeletons, necropsy findings or reproductive functions. No adverse effects of S-1 on F2 offspring were found in any treatment groups. Under the conditions of the present study, the non-observed effect dose levels of S-1 was 4 mg/kg/day for general toxicology of dams, 7 mg/kg/day for reproductive ability of dams, 1 mg/kg/day for postnatal growth in F1 offspring and 7 mg/kg/day for postnatal growth in F2 offspring.

Diethylstilbestrol-Induced Perinatal Lethality in the Rat. I. Relationship to Reduced Maternal Weight Gain1

An analysis was performed to determine the mechanism of depressed maternal weight gain and its effect on perinatal lethality following prenatal exposure to diethylstilbestrol (DES). Pregnant Sprague-Dawley rats were dosed orally by gavage with DES or corn oil (control) during various intervals of gestation. The maternal weight-gain patterns of control and treated dams and the number of live offspring were recorded. The amounts of feed and water intake and feces and urine output in pregnant dams were measured, and metabolic rate and thyroid hormone levels were also determined. DES (at 45 micrograms/kg/day) was embryo- and fetolethal during implantation and parturition, and there was an accompanying decline in maternal weight. Growth of adult males, nonpregnant females, and weanlings of both sexes was also depressed. During pregnancy, the net intake of feed and water was not altered by the drug, but maternal serum thyroxine and metabolic rate were significantly elevated. Reduced metabolic efficiency, then, is the likely mechanism for weight depression. Reduction of maternal weight gain during pregnancy by DES is a diagnostic indicator of fetolethality, but is probably not causally related to it.

Toxic effects of sodium selenite on pregnant mice and modification of the effects by vitamin E or reduced glutathione.

Toxic effects of sodium selenite (SS) on dams and/or conceptuses and modification of the effects by vitamin E (VE) or reduced glutathione (GSH) were investigated in pregnant mice during late gestation. Dose-dependent fetocidal effects and fetal growth retardation were observed in mothers injected SC with SS on day 12 but not on day 16 of gestation. On day 12, abortion occurred only at a dose level of 58.8 mumol/kg SS, whereas on day 16 it occurred at two dose levels: 27.2 and 40.0 mumol/kg SS. Pregnant mice were treated with or without VE (50 mg/kg, SC, daily from day 7 to day 11) and injected with a single dose of SS (46.2 mumol/kg, SC) on day 12. VE prevented abortion and maternal death, but none of the other effects such as fetal loss, fetal growth retardation, and decrease in body weight gain of the dams. Pretreatment with GSH (0, 2, or 5 mM/kg, SC) 20 min before SS injection on day 12 exacerbated maternal death, abortion, and decrease in body weight gain of the dams. No effect were observed in the number of live offspring, and in body weight of offspring.

Family size and evolution of Rh polymorphism

A reproductive compensation model has been proposed, where each family produces exactly the same number of live offspring. Under that condition, the frequency of d alleles in the Rh blood group always increases. This would account for populations with high frequency of d alleles.