Improved sperm cryosurvival in diluents containing amides versus glycerol in the Przewalski's horse (Equus ferus przewalskii)

Two studies were conducted to understand sperm cryosensitivity in an endangered equid, the Przewalski's horse (Equus ferns przewalski), while testing the cryoprotectant ability of formamides. The first assessed the toxicity of permeating cryoprotectants (glycerol, methylformamide IMF] and dimethylformamide [DMF]) to Przewalski's horse spermatozoa during liquid storage at 4 C. The second examined the comparative influence of three diluents (with or without formamides) on cryosurvival of sperm from the Przewalski's versus domestic horse. When Przewalski's horse spermatozoa were incubated at 4 C in INRA 96 with differing concentrations of glycerol, MF or DMF or a combination of these amides, cells tolerated all but the highest concentration (10% v/v) of MF alone or in combination with DMF, both of which decreased (P < 0.05) motility traits. There was no effect of cryoprotectants on sperm acrosomal integrity. In the cryosurvival study, average sperm motility and proportion of cells with intact acrosomes in fresh ejaculates were similar (P> 0.05) between the Przewalski's (67%, 84%, respectively) and domestic (66%, 76%) horse donors. Sperm from both species were diluted in lactose-EDTA-glycerol (EQ), Botu-Crio (BOTU; a proprietary product containing glycerol and MF) or SM (INRA 96 plus 2% [v/v] egg yolk and 2.5% [v/v] MF and DMF) and then frozen over liquid nitrogen vapor. After thawing, the highest values recovered for total and progressive sperm motility, acrosomal integrity and mitochondria] membrane potential were 42.4%, 21.8%, 88.7% and 25.4 CN (CN = mean JC-1 fluorescence intensity/cell on a channel number scale), respectively, in the Przewalski's and 49.3%, 24.6%, 88.9% and 25.8 CN, respectively, in the domestic horse. Although sperm progressive motility and acrosome integrity did not differ (P> 0.05) among treatments across species, mitochondrial membrane potential was higher (P< 0.05) in both species using EQ compared to BOTU or SM media. Additionally, Przewalski's stallion sperm expressed higher (P < 0.05) post-thaw total motility in BOTU and SM compared to EQ whereas there were no differences among freezing diluents in the domestic horse. In summary, Przewalski's stallion sperm benefit from exposure to either MF or DMF as an alternative cryoprotectant to glycerol. Overt sperm quality appears similar between the Przewalski's and domestic horse, although the total motility of cells from the former appears more sensitive to certain freezing diluents. Nonetheless, post-thaw motility and acrosomal integrity values for Przewalski's horse spermatozoa mimic findings in the domestic horse in the presence of INRA 96 supplemented with 2% (v/v) egg yolk and a combined 2.5% concentration of MF and DMF. Published by Elsevier Inc.

Sperm fertility and viability following 48 h of refrigeration: Evaluation of different extenders for the preservation of bull semen in liquid state

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

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.

Values of 13,14-Dihydro-15-Keto-PGF(2 alpha), Progesterone and Oestradiol After Injection of Prostaglandin at Different Periods of Buffalo Puerperium

Knowledge of the effectiveness of prostaglandins in uterine involution process led to the development of protocols with its analogues in postpartum period. However, this hormone mechanism of action is not yet fully elucidated. Thus, the objective of this study was to verify if chloprostenol administration, at early or intermediary puerperium, can induce changes on progesterone, PGFM and oestradiol plasma concentrations. 30 Murrah postpartum buffaloes were randomly divided into three groups: CONT (saline, n = 10); CLO2 (chloprostenol at days 2 and 5 postpartum, n = 10) and; CLO15 (chloprostenol at days 15 and 20 postpartum, n = 10). Blood samples were collected from jugular vein to measure progesterone, PGFM and oestradiol plasma concentrations at days 2, 7, 14, 21 and 28 postpartum. CLO2 group presented lower progesterone and PGFM plasma concentrations in relation to CONT and CLO15 groups (0.23 +/- 0.00 and 0.32 +/- 0.11, 0.19 +/- 0.00 and 0.23 +/- 0.11, 0.23 +/- 0.00 and 0.30 +/- 0.19, for groups CONT, CLO2 and CLO15, respectively; P < 0.05). There was no significant difference in oestradiol plasma concentration between experimental groups (P > 0.05). Prostaglandin synthetic analogue administration induced hormonal changes in postpartum buffaloes, which can partially explain its positive effect under reproductive function of this specie.

Comparison of Botu-bov and Tris as Freezing Extenders of Buffalo Sperm Recovered from Epididymal Cauda

Valuable genetic material can be preserved by the cryopreservation of epididymal sperm. This study evaluated the viability of pre-freezing and post-thawed sperm samples recovered from the epididymal cauda of buffaloes. Epididymides from eight Murrah buffaloes with 18 months of age were used. Semen samples were diluted in two different freezing extenders: Botu-bov (BB) and Tris (TRIS). Immediately after slaughter, both testicles from each animal were collected and transported at 4 degrees C for six hours interval. In laboratory, the removed epididymides were flushed to obtain sperm and the fractions were diluted in both freezing extenders (BB and TRIS). Semen doses were analyzed before and after frozen at -196 degrees C. BB and TRIS pre-freezing results were 38.54 +/- 22.33%(b) and 14.17 +/- 12.78%(a) for total motility (TM), 25.00 +/- 16.12(a) and 9.44 +/- 9.11(a) for progressive motility (PM), 7.21 +/- 0.98(a) and 5.09 +/- 2.65(a) for percentage of rapid cells (RAP), 91.08 +/- 12.53(b) and 63.33 +/- 31.47(a) for velocity of trajectory (VAP), 73.54 +/- 20.17(b) and 49.50 +/- 9.11(a) for linear progressive velocity (VSL), 172.21 +/- 24.55(a) and 116.94 +/- 59.48(a) for curvilinear velocity (VCL), respectively (P < 0.05). BB and TRIS post-thawing results were 42.25 +/- 21.50(b) and 17.62 +/- 19.46(a) for TM, 27.25 +/- 24.86(a) and 18.00 +/- 13.68(a) for PM, 7.35 +/- 0.98(a) and 6.26 +/- 1.13(a) for RAP, 91.42 +/- 16.86(a) and 75.96 +/- 13.17(a) for VAP, 67.96 +/- 12.13(a) and 60.04 +/- 10.42(a) for VSL, 177.54 +/- 23.53(b) and 141.29 +/- 24.97a for VCL, respectively (P < 0.05). The sperm recovered from the epididymal cauda, after 6 h storage of epididymides at 5 degrees C ensures sperm preservation demonstrating that the diluent Botu-bov had higher total motility both pre-and post-freezing when compared with TRIS. Additionally, the sperm frozen with the diluent Botu-bov showed higher values of VSL at post-thawing. These findings may reflect in improvement of conception rates.

Adding Motility Stimulants to Improve Freezing of Buffalo Sperm Recovered from Epididymal Cauda

The cryopreservation of epididymal sperm is important to preserve genetic material from valuable buffalo bulls. This study evaluated the viability of post-thawed sperm samples recovered from the epididymal cauda adding motility inductors. For that, were used epididymides from eight Murrah buffaloes with 18 months of age. Semen samples were submitted to three different conditions: (CT - control) without adding medium, (SPERM) adding Sperm Talp medium, and (FERT) adding Fert Talp medium. Immediately after slaughter, both testicles from each animal were collected and transported at 4 degrees C at maximum six hours interval. In laboratory, the removed epididymides was flushed to obtain sperm and diluted in the freezing extender. Each buffalo sperm were divided and fractions were submitted to all conditions (CT, SPERM and FERT). Semen doses were frozen at -196 degrees C. CT, SPERM and FERT post-thawing results were 13.63 +/- 8.91, 38.77 +/- 8.91 and 42.83 +/- 8.91 for total motility, 7.30 +/- 8.74, 24.87 +/- 8.74 and 29.70 +/- 8.74 for progressive motility, 6.04 +/- 0.92, 6.74 +/- 0.92 and 6.93 +/- 0.92 for percentage of rapid cells (P < 0.05). In conclusion, diluted semen supplementation with Sperm or Fert talp increases the motility of cauda epididymal sperm of buffalo bulls.

Uterine Involution, Fluid Accumulation and Ovarian Activity after Injection of Prostaglandin at Different Periods of Buffalo Puerperium

This study aimed to evaluate the effect of chloprostenol administration, at early or intermediary puerperium, under uterine involution, intrauterine fluid accumulation and ovarian activity return. 30 Murrah postpartum buffaloes were randomly divided into three groups: CONT (saline, n = 10); CLO2 (chloprostenol at days 2 and 5 postpartum, n = 10) and; CLO15 (chloprostenol at days 15 and 20 postpartum, n = 10). Gynecological exams were performed at days 2, 7, 14, 21 and 28 postpartum, when uterine involution degree (1 to 3 scale, by transrectal palpation), intrauterine fluid accumulation (0 to 3 scale, by ultrasound exam) and ovarian activity (B-mode ultrasound exam) were evaluated. CLO2 group presented higher uterine involution (2.00 +/- 0.23, 1.66 +/- 0.23, 1.58 +/- 0.23 for groups CLO2, CONT and CLO15, respectively) and faster ovarian activity return in relation to groups CONT and CLO15 (P < 0.05). Groups CLO2 and CLO15 showed lower intrauterine fluid accumulation compared to CONT group (2.04 +/- 0.20, 1.58 +/- 0.20, 1.92 +/- 0.20 for groups CONT, CLO2 and CLO15, respectively; P < 0.05). Prostaglandin analogue administration in postpartum buffalo benefited uterine involution, lochia expulsion and ovarian activity return, improving reproductive efficiency in this specie.

Sperm Bundles in the Seminal Vesicle of the Crematogaster victima (Smith) Adult Males (Hymenoptera: Formicidae)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Assessment of in vitro sperm characteristics and their importance in the prediction of conception rate in a bovine timed-AI program

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Exploratory analysis of differences in sperm morphology in Nelore and Gir (Bos indicus) bulls

The aim of this study was to evaluate the effects of breed and season on semen quality parameters of zebu bulls. Data (1,632 registers) of semen production from Gir (n = 4) and Nelore (n = 15) bulls were collected between October 2005 and November 2009. The ejaculates were collected twice a week during various seasons (summer, fall, winter, and spring) and evaluated for the following semen parameters: ejaculate volume, sperm concentration, sperm motility, forward progressive motility (FPM), and sperm morphology. Factor analysis was used to determine the relationship among variables. The effect of breed (Gir and Nelore) and season and their cross effect on each parameter and extracted factor were tested using ANOVA. A negative correlation (P < 0.05) was observed between FPM and proximal droplet, as well as with abnormal loose head, abnormal small head, pouch formation, abnormal mid-peace, and strongly folded tail. Gir bull sperm showed more major defects, detached acrosome, and minor FPM (P < 0.01), whereas Nelore bulls showed a higher number of sperm with normally loose head.