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Precise pregnancy detection is critical for optimizing reproductive efficiency in animals. In mice, the conventional copulation plug method for pregnancy confirmation lacks reliability, particularly in inbred strains, with false-pregnancy rates exceeding 50 %. Here, to improve pregnancy detection efficiency in mice, we examined vaginal cell population changes using vaginal smear technique and analyzed MMP-9 expression in vaginal tract from day 0.5 to 6.5 of pregnancy. A total of forty-four female mice in the estrus stage were paired with sexually mature male mice for natural mating. The morning after mating was considered as embryonic day 0.5 (E0.5). Vaginal cytology was used to determine the vaginal cells of pregnant and non-pregnant mice from day 0.5 to 6.5. Furthermore, the protein and gene expression of MMP-9 were assessed through the utilization of immunohistochemistry and real-time PCR. Our findings reveal that pregnant mice display the higher number of leukocytes but the low number of nucleated/cornified epithelial cells in vagina compared to the non-pregnant counterparts. Furthermore, MMP-9 protein consistently presents in vaginal epithelial cells throughout the experimental period, with increased protein expression on day 4.5 of pregnancy followed by a decline on day 6.5 of pregnancy. This result is similar to MMP-9 gene expression, reaching the peak on day 5.5 of pregnancy. In conclusion, vaginal cytology proves effective for mouse pregnancy determination, while MMP-9 can serve as a potential biomarker for detecting pregnancy in mice.

There has been limited work done on estrus synchronization or induction in the Dasyprocta species. This experiment sought to apply three different methods of estrus synchronization to fifteen agoutis (Dasyprocta leporina) which were placed into three treatments. Treatment 1 received gonadorelin acetate (Fertiline®) injections subcutaneously (40 µg); treatment 2 received two doses of dinoprost tromethamine (Lutalyse®) injections intramuscularly (2.5 mg) nine days apart and; treatment 3 received gonadorelin acetate (Fertiline®) injections subcutaneously (40 µg), followed seven days later by dinoprost tromethamine (Lutalyse®) injections intramuscularly (2.5 mg) and a second gonadorelin acetate (Fertiline®) injection subcutaneously (40 µg) given two days after. Daily vaginal swabbing, followed by cytological analysis was done until estrus was noted in all animals. All animals were successfully synchronized, with estrus being observed 17.40 ± 0.24, 7.25 ± 0.25 and 7.60 ± 0.24 days post treatment for treatments 1, 2 and 3, respectively. Return to estrus in females from treatment 1 was later than the other two treatments (p

abstract: Initiating a breeding program using animals sourced from the wild can be challenging due to the time it takes to determine their basic reproductive traits in the ex situ environment. Often, the needs for such a program arise rapidly due to a catastrophic event. After multiple wildfires decimated much of the endangered Mt. Graham red squirrel’s (Tamiasciurus fremonti grahamensis) limited habitat, the U.S. Fish and Wildlife Service and the Phoenix Zoo (Phoenix, Arizona, USA) initiated a pilot breeding program using wild-sourced individuals, including the five (three males, two females) studied here. To inform the breeding program, we sought to characterize the reproductive biology of the Mt. Graham red squirrel by developing non-invasive and/or or minimally invasive methods. Our specific objectives were to: 1) characterize reproductive biology via fecal hormone metabolite analysis; 2) determine how reproductive hormones vary across the breeding season; 3) determine the relationship between the hormonal analysis and the physiological changes, and 4) identify a non-invasive or minimally invasive method to rapidly assess when females are close to ovulation. Results showed that follicular phases (number of days fecal estrogen metabolites (FEMs) were elevated) lasted ~26 days (range, 14–51 days). The only female to ovulate ovulated three times with fecal progestogen metabolites (FPMs) remaining elevated for ~21 days. Peak vulvar swelling in this female was associated with higher FEM and the peri-ovulatory period, but vaginal cytology results were less useful for predicting ovulation. For males, scrotum enlargement occurred in January and February and fecal androgen metabolites (FAMs) were elevated in March and June, with lowest concentrations in August, October, and December. Although no successful copulation was observed among paired squirrels, this study provides valuable information about Mt. Graham red squirrel reproductive biology and establishes non-invasive methods that can be used to determine reproductive readiness and make ex situ management decisions. Furthermore, this ex situ study is the first to characterize estrogen and progesterone metabolite patterns for an American tree squirrel (Tribe: Sciurini) across the breeding season.

Background: Women with human immunodeficiency virus have higher rates of abnormal cervical and vaginal cytology and, subsequently, of cervical and vaginal cancers. Although professional bodies currently advocate for indefinite cytology screening for women living with human immunodeficiency virus, these recommendations are based on expert opinion, not evidence-based. In the general population, women who have never had an abnormal cytology result can cease screening at age 65 years. This is due to the relatively low incidence of dysplasia in this group and the risk of false-positive results as women age, invasive follow-up testing, and destructive treatments of lesions that are unlikely to progress to cancer. What is unclear, however, is how human immunodeficiency virus-infected women over age 65 years who have no history of abnormal cytology should be screened to maximize benefit while reducing harms of overscreening. This is a crucial question, as women over age 65 years who are living with human immunodeficiency virus comprise a rapidly growing population., Objective: To describe the incidence of abnormal cervical and vaginal cytology results in women over the age of 65 years living with human immunodeficiency virus, with the goal of providing evidence for screening recommendations., Materials and Methods: A retrospective chart review was performed, identifying 69 women who received gynecologic follow-up in a county hospital system in Houston, Texas, between 2000 and 2018 and who met study criteria. Incidence of abnormal cytology after age 65 was determined by analyzing all available cytology results after age 65. Demographic and clinical risk factors, including human immunodeficiency virus-specific clinical risk factors, were analyzed. Matched cervical and vaginal pathology results, if conducted, were also evaluated. Statistical analyses were conducted using Stata 15, including χ 2 tests and Wilcoxon rank-sum tests for categorical and continuous variables, respectively. Estimates of the cumulative probability of developing an abnormal cytology result was calculated using the Kaplan-Meier method., Results: Among 69 women with no history of abnormal cervical cytology, 12 (17%) went on to develop abnormal cytology results, including 3 (4%) showing high-grade squamous intraepithelial lesions. The incidence rate was 3.5 cases per 100 woman-years (95% confidence interval, 1.58, 7.81). No demographic or gynecologic characteristics were associated with abnormal cytology. A CD4 count of <200 at the time of human immunodeficiency virus diagnosis or at the time of cytology was associated with an abnormal Papanicolaou test result (P < .0001, P = .031). Of women with pathology results in the county hospital system (n = 8), 4 (50%) had cervical intraepithelial neoplasia 2+ or vaginal intraepithelial neoplasia 2+. No women developed invasive cancer. However, 50% of women who had an abnormal Papanicolaou test result in the study period were lost to follow-up; outcomes for these patients are unknown., Conclusion: Given the relatively high proportion (4%) of women with high-grade squamous intraepithelial lesions/cervical intraepithelial neoplasia 2+/vaginal intraepithelial neoplasia 2+ during the study period, we agree with current screening recommendations for continued routine Papanicolaou testing after the age of 65 years in women with human immunodeficiency virus. More evidence from larger studies is needed to solidify evidence-based screening recommendations in this unique and growing population., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Determination of the timing of the estrus cycle is essential for fertile mating. There are physiological variations among breeds, between bitches, and between cycles of the same bitch. If serial monitoring and many tools are applied, the exact moment of ovulation could be pin-pointed. However, it leads to time and costly difficulties. Progesterone concentrations during estrus cycles follow a specific pattern, and it is largely used in timing of fertile period. Although it has similar pattern in general, it is likely that breed-specific differences exist. The aim of this study was to investigate the way of timing the fertile period for successful pregnancy in American Bully dogs based on vaginal cytology and progesterone assay with minimized cost. To identify the empirical relations among reproductive characteristics, we performed statistical analyses on data from proestrus-to-estrus 27 American Bully dogs referred for 7 months. We found the significant correlations between the cyclic changes of vaginal cytology and progesterone assay. The relationship of serum progesterone concentrations with the days from vaginal discharge onset was analyzed through linear regression assay. In conclusion, we addressed two standards in the timing of fertile period with a minimal number of progesterone assays in the breeding management of American Bully dogs., (Copyright © 2017. Published by Elsevier Inc.)

The goal was to increase pseudopregnant mice production by estrous cycle staging by visual examination before pairing and to determine the effect of such pseudopregnant recipients on embryo transfer. To compare methods of estrous cycle staging over 14 days, groups consisted of 10 females in proestrus-estrus and 10 vasectomized males; group 1: only daily visual observation; group 2: daily visual observation and cytological examination on day 1; group 3: daily visual observation and daily cytological examination. The average time to first vaginal plug was 1.8 days in group 1, 2.7 days in group 2, and 3.2 days in group 3, whereas the average time between consecutive vaginal plugs was 9.2 days (group 1), 10 days (group 2), and 9.25 days (group 3). The average time between consecutive estrous cycles was 9.7 days (group 1), 11.8 days (group 2), and 9.4 days (group 3). The congruence between visual and cytological examination in determining proestrus-estrus in group 2 was 100% and that for the four stages in group 3 was 79% with a range of 44% to 100%. From 162 plug-positive females originally selected in proestrus-estrus, 49%, 30%, 19%, and 2% were plug-positive on Day 1, Day 2, Day 3, and Day 4, respectively, showing that pseudopregnant mice production was significantly increased on the first 2 days. From 192 plug-positive females originally selected randomly, these values were 31%, 21%, 35%, 10%, and 3% on d1, d2, d3, d4, and d5, respectively. No significant differences were observed between groups with respect to embryo transfers with fresh or cryopreserved embryos although the number of pups born per litter was higher in group A with fresh (7.57 vs. 6.39) and cryopreserved-thawed embryos (5.0 vs. 4.38). Furthermore, the sex ratio and the genotype of the pups were not significantly affected., (Copyright © 2016 Elsevier Inc. All rights reserved.)