Use of Infrared Thermography during Ejaculation Process and Its Link with Semen Quality and Freezability in Dogs.

This study aimed to describe the thermal variation of external reproductive tracts during ejaculation in relation to sperm quality in dogs. Forty-six adult fertile dogs were monitored using a thermal camera before, during and after the semen collection, taking into account penile and scrotal temperatures as reproductive thermal patterns while eye and perianal temperatures were recorded as complementary thermal patterns of behavioral response. The parameters were classified depending on age (≤4 years and >4 years), body weight (BW) (≤75 kg and >75 kg), sperm concentration (CON) (≤300 million and >300 million), total testicular volume (TTV) (≤600 cm3 and >600 cm3) and total ejaculation time (TET) (≤800 s and >800 s) of the animals from which semen was collected successfully. Heavier males (p < 0.05) that have more consistent testicles (p < 0.01) as well as quicker ejaculate responders (p < 0.001) and lower scrotal temperature had better semen (Δ motility) freezability. The lower eye temperature prior to the ejaculation (p < 0.01), lower scrotal temperature following ejaculation (p < 0.01), and conversely, higher penile temperature during the ejaculation (p < 0.001) had a higher sperm concentration. Furthermore, the sperm freezability was negatively correlated with total ejaculation time (r = -0.39, p < 0.05) and sperm abnormalities were lower in the ejaculate of dogs having a higher temperature of the scrotum, bulbus and penis. In conclusion, infrared monitoring throughout semen collection in dogs can provide information on behavioral reactions during human manipulation, as well as semen quality and testicular functionality.

Semen collection by trans-rectal digital stimulation and insemination campaign in goat.

The overall purpose of this study was to describe a method of semen collection via trans-rectal digital massage (TDM) and to carry out a related fertility trial in Angora goat. Sixteen Angora bucks (ranging 1-4 years) and 28 nulliparous does (1-2 years) were used in this study. Semen samples were collected via trans-rectal massage from 85.71% of the bucks in multiple attempts (18/21). The mean values of volume, pH, mass motility, total motility, concentration, viability, abnormal spermatozoa rate and ejaculation time were 0.64 ± 0.09 ml, 6.3 ± 0.21, 2.7 ± 0.34, 58.18 ± 5.1%, 3.68 ± 0.31 × 109 /ml, 71.38 ± 7.12%, 18.22 ± 2.48% and 3.4 ± 0.33 min respectively. Oestrus was detected with teaser buck and confirmed by using infrared thermography and ultrasonography (US). The success rate of synchronisation was found as 71.4% (20/28). On Day 21, pregnancy diagnosis was performed trans-rectally with US and the pregnancy rate was determined as 78.57% (11/14). TDM method of semen collection seems to be easily applicable to the buck and it could be a good alternative to collect semen as well as its use in artificial insemination campaign. Thermal monitoring is found to be a valuable tool to monitor the response to hormonal driven ovulatory synchronisation in Angora does during timed artificial insemination.

Effects of phthalates on bovine primary testicular culture and spermatozoa.

Among environmental endocrine-active chemicals, phthalates, commonly known as plasticizers, disrupt the development of the male reproductive tract. In this study, the effects of phthalates (DIBP, BBP, DINP, DBP, DEP, DEHP and DMP) were evaluated on cultures of bovine primary male reproductive cells (n = 3) and spermatozoa (n = 4). Epididymal (caput and corpus epididymis), testicular (parenchymal and mediastinal/tubular) and vas deferens cells (VDC) were prepared from samples collected from slaughterhouse. Second part of caput epididymis which have fewer amount of principal cells, were found to be less affected compared to the first part except DEHP; while corpus epididymis was found to be more affected with IC50 values below 0.976 ng/mL (except for DEP at 4.97 ng/mL). In testicular parenchymal cells, IC50 ranged from 0.15 to 4.11 ng/mL and for mediastinum from 0.01 to 7.31 ng/mL; where cytotoxic effects were more evident in mediastinal section. Least cytotoxic and even proliferational effects (DEHP, DMP and DEP) were observed in VDC, the muscular tube carrying sperm from epididymis to the ejaculatory duct. Least spermiotoxic phthalate was DBP (3.928 ng/mL); while DINP (0.550 ng/mL) induced highest cytotoxic effect on bovine spermatozoa. Differences in the cellular structure and/or the androgen receptor distribution effect the toxicity of phthalates. Our preliminary findings on bovine spermatozoa indicate possible morphological and motility alterations; which challenges further investigation of the transition of phthalates on semen straws used in cryopreservation. Increase of exposure to environmental contaminants raise the issue of the requirement of a new perspective on reproductive health, species and tissue specific differences should further be emphasized.

Effect of polyvinyl alcohol on survival and function of angora buck spermatozoa following cryopreservation.

The aim of this study was to examine effects of polyvinyl alcohol (PVA) on buck semen quality. Seventy-five ejaculates were collected and diluted in Tris-egg yolk extender containing one of three PVA co-polymers of 9, 18 and 100 kDa. Five different concentrations 0.001, 0.01, 0.1, 1 and 2% of the PVA co-polymers were added to the extender with respected to the decreasing glycerol concentrations of 5, 4, 3, 3, 2% respectively. Following freeze-thaw, sperm motility, viability, acrosome-intact spermatozoa and mitochondrial membrane potentials were analysed. During freezing, sperm seeding temperature were recorded with a cryo-thermometer. PVA 2% glycerol group gained 8.2 ±â€¯1 °C latent heat plateau difference comparing to control. Highest motility was found in PVA 18 kDa with regardless of the dosage (P < 0.001). All PVA copolymers gained higher motility independently in all other dosage groups (except PVA 2%) comparing to control (P < 0.001). Live spermatozoa rate between treatment groups were statistically insignificant (P = 0.953), however, when moribund sperm were gated out PVA 9 induced better protection with respect to other groups (P < 0.05). Intact acrosome rate was statically higher in PVA groups (P < 0.002) and subgroups (P < 0.001). Mitochondrial membrane potential was higher in all experimental groups comparing to control group (P < 0.001). PVA co-polymer concentrations of 0.01, 0.1, 1 and 2% v/v (PBS: PVA) decreased the concentration of glycerol required for freezing in a 100 ml volume by 0, 1, 2, 2, and 3% v/v from the control dose (5%), respectively. In conclusion, synthetic PVA-derived ice blocking agents offer new opportunities for improving the post-thaw buck sperm quality.

The use of infrared thermography to detect the stages of estrus cycle and ovulation time in anatolian shepherd dogs.

BACKGROUND: The aim of the study is to evaluate the effectiveness of thermographic monitoring, using the temperature changes of perianal and perivulvar areas for the determination of estrus in Anatolian Shepherd bitches. Fifteen bitches were used in the study. Blood and vaginal smear samples were collected and thermographic monitoring of perianal and perivulvar areas were carried out starting from proestrus to early diestrus. Also, external signs of estrus were investigated. Smear samples were evaluated by light microscopy after Diff-Quik staining method and superficial and keratinized superficial cells were determined as percentage (S + KS%). Progesterone and luteinizing hormone measurements were done by radioimmunoassay. The difference in temperature between perianal and perivulvar areas was evaluated through thermographic images by FLIR ResearchIR Software. RESULTS: According to the results obtained from the study, differences between progesterone and S + KS% were statistically significant (P < 0,05). Although temperature showed increase and decrease with progesterone and S + KS%, the differences were not important statistically (P > 0,05). Serum luteinizing hormone levels did not sign any difference (P > 0,05). CONCLUSIONS: As a result, thermographic monitoring alone is not enough for estrus detection in Anatolian Shepherd bitches. However, it can be used to assist the actual estrus detection technique in terms of providing some foreknowledge by evaluating the differences in temperature.

Vulvar thermal pattern following synchronization of estrus is linked to fertility after timed artificial insemination in goat.

Aim of this study was to study the vulvar thermal pattern variation during the timed artificial insemination protocol in Angora goat and identify the relationship with the successful rate. Does (36 adult healthy females) were synchronized using PGF2α at the day 0, 11 days of progesterone impregnated sponges intra-vaginally, PMSG 48 h before sponges withdraw (day 11) and the intra-cervical inseminations were carried out 48 h later (Timed Artificial Insemination: TAI) with chilled semen. Vulvar (VST) and perivulvar (PST) areas were considered to evaluate the thermal pattern during the protocol at the day 0 and at the TAI using a thermo camera (E60, FLIR System). Differences of temperature (ΔT) between the surfaces were calculated for each time. The does were monitored for pregnancy, delivery time and prolificacy. Pregnant (P) and non-pregnant (NP) does were compared in terms of VST, PST and ΔT using two ways ANOVA considering time and pregnancy as sources of variability. VST was lower than PST in all the monitored does (P < 0.05) (34.79 ± 0.14 vs 36.58 ± 0.14 °C) and without differences between P and NP at day 0 (35 ± 0.18 vs 36.39 ± 0.22 °C). Significant difference (P < 0.05) between P and NP does was recorded at TAI in terms of VST (33.89 ± 0.31 vs 35.40 ± 0.24 °C) and ΔT (-3.16 ± 0.34 vs -1.62 ± 0.26 °C). In conclusion thermal emission by glabrous surfaces in goat may be used to identify the right response induced by hormonal treatments and to optimize the application of assisted reproductive techniques at the field level.