Evaluation of sperm quality in successive regular collections from captive black-and-gold howler monkeys (Alouatta caraya)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Differential distribution of sperm subpopulations and incidence of pleiomorphisms in ejaculates of captive howling monkeys (Alouatta caraya)

The aim of this study was to develop an objective method to determine the incidence of pleiomorphisms and its influence on the distribution of sperm morphometric subpopulations in ejaculates of howling monkeys (Alouatta caraya) by using a combination of computerized analysis system (ASMA) and principal component analysis (PCA) methods. Ejaculates were collected by electroejaculation methods on a regular basis from five individuals maintained under identical captive environmental, nutritional, and management conditions. Each sperm head was measured for dimensional parameters (Area [A, (square micrometers)], Perimeter [P, (micrometers)], Length [L, (micrometers)], and Width [W, (micrometers)]) and shape-derived parameters (Ellipticity [(L/W)], Elongation [(L - W)/(L + W)], and Rugosity [(4 pi A/P-2)]). PCA revealed two principal components explaining more than the 96 % of the variance. Clustering methods and discriminant analyzes were performed and seven separate subpopulations were identified. There were differences (P < 0.001) in the distribution of the seven subpopulations as well as in the incidence of abnormal pleiomorphisms (58.6 %, 49.8 %, 35.1 %, 66.4 %, and 55.1 %, P < 0.05) among the five donors tested. Our results indicated that differences among individuals related to the incidence of pleiomorphisms, and sperm subpopulational structure was not related to the captivity conditions or the sperm collection method, since all individuals were studied under identical conditions. In conclusion, the combination of ASMA and PCA is a useful clinical diagnostic resource for detecting deficiencies in sperm morphology and sperm subpopulations in A. caraya ejaculates that could be used in ex situ conservation programs of threatened species in Alouatta genus or even other endangered neotropical primate species.

Efficacy of sperm motility after processing and incubation to predict pregnancy after intrauterine insemination in normospermic individuals

Background: Intrauterine insemination (IUI) is widely used to treat infertility, and its adequate indication is important to obtain good pregnancy rates. To assess which couples could benefit from IUI, this study aimed to evaluate whether sperm motility using a discontinuous gradient of different densities and incubation in CO2 in normospermic individuals is able to predict pregnancy.Methods: A total of 175 couples underwent 175 IUI cycles. The inclusion criteria for women were as follows: 35 years old or younger (age range: from 27 to 35 years) with normal fallopian tubes; endometriosis grades I-II; unexplained infertility; nonhyperandrogenic ovulatory dysfunction. Men with normal seminal parameters were also included. All patients underwent ovarian stimulation with clomiphene citrate and human hMG or r-FSH. When one or (at most) three follicles measuring 18 to 20 mm were observed, hCG (5000 UI) or r-hCG (250 mcg) was administered and IUI performed 36-40 h after hCG. Sperm processing was performed using a discontinuous concentration gradient. A 20 microliters aliquot was incubated for 24 h at 37 degrees C in 5% CO2 following a total progressive motility analysis. The Mann-Whitney and Chi-square tests, as well as a ROC curve were used to determine the cutoff value for motility.Results: Of the 175 couples, 52 (in 52 IUI cycles) achieved clinical pregnancies (CP rate per cycle: 29.7%). The analysis of age, duration and causes of infertility did not indicate any statistical significance between pregnancy and no pregnancy groups, similar to the results for total sperm count and morphology analyses, excluding progressive motility (p < 0.0001). The comparison of progressive motility after processing and 24 h after incubation between these two groups indicated that progressive motility 24 h after incubation was higher in the pregnancy group. The analysis of the progressive motility of the pregnancy group after processing and 24 h after incubation has not shown any motility difference at 24 h after incubation; additionally, in couples who did not obtain pregnancy, there was a statistically significant decrease in progressive motility 24 h after incubation (p < 0.0001). The ROC curve analysis generated a cutoff value of 56.5% for progressive motility at 24 h after incubation and this cutoff value produced 96.1% sensitivity, 92.7% specificity, 84.7% positive predictive value and 98.3% negative predictive value.Conclusions: We concluded that the sperm motility of normospermic individuals 24 h after incubation at 37 degrees C in 5% CO2, with a cutoff value of 56.5%, is predictive of IUI success.

Effect of testicular thermoregulation on the quality of buffalo sperm

The process of spermatic division and differentiation (spermatogenesis) occurs with intratesticular temperature lower that the corporal temperature and for that is essential that the testicular thermoregulation mechanism occurs properly. For evaluation of the scrotal surface temperature can be used the infrared thermography or testicular sensors, besides that, can be evaluated the blood flux in the spermatic cord through the Doppler ultrasonography. Thus, the aim of this study is to analyze the testicular thermoregulation in adult buffaloes through scrotal thermography and Doppler ultrasound of testicular artery and verify its effect on sperm quality. For that were used seven healthy buffaloes, with age of 3 and 4 years, of the Murrah breed. The animals were subjected to 3 semen collections using artificial vagina, with one day of interval. In addiction, the retal temperature measurement (RT) with dry bulb thermometer, the measurement of scrotal surface temperature (SST) and body surface temperature (BST) through infrared thermography and the pulsatility (PI) and resistivity (RI) index of testicular artery by Doppler ultrasonography, were performed using 2 distinct moments: animals previously placed to shade (M1) and animals subjected to 4 hours of sun (M2). All parameters were compared by T test and the correlations were performed by Pearson test using the In Stat Graph Pad 3 (R) program. The significant level considered was 5%. There was an increase (p<0,05) of RT, SST, SNT and RI in M2. increasing trend was observed (0,05>p>0,01) PI and RI between M1 and M2. There was a low correlation between SST and semen quality. The results of this study allow us to conclude that adult buffaloes have low ability to perform body and testicular thermoregulation in situations of enviromental heat stress. However, this low capacity of testicular temperature maintenance demonstrated no correlation with the sperm kinetic parameters and sperm morphological defects in buffalo spermatozoa.