Improved the in vitro blastocyst development in oocytes recovered from cyclic buffaloes.

This study examined the impact of cyclicity (with or without cycle corpus luteum; CL) on oocyte quality and embryonic development in buffaloes. We collected oocytes from the ovaries of slaughtered buffaloes (N = 158 cyclic; n = 316 ovaries and N = 177 acyclic; n = 353 ovaries). Blood progesterone concentration and number of oocytes per ovary were higher in cyclic buffaloes. Cyclic buffalo ovaries produce higher oocytes with I + II and fewer III + IV grades. Oocytes from cyclic buffaloes had a higher maturation rate based on cumulus expansion, cleavage rate and embryo development to the 8-cell, morula and blastocyst stages than acyclic buffaloes. In conclusion, oocytes recovered from the ovaries of the cyclic buffaloes showed improved oocyte competence and subsequent in vitro blastocyst development.

Exploring testicular miRNA profiles during developmental stages in Dezhou donkeys: A preliminary insight.

Testicular development and spermatogenesis are complex phenomena controlled by various genetic factors, including miRNA-based post-transcriptional gene expression regulation. Exploring the miRNA expression patterns during testicular development in Dezhou donkeys would enhance our understanding of equine fertility and spermatogenesis. In this investigation, we examined the testicular miRNA profiles at various stages of development. The experimental animals were divided into three groups based on their developmental stages: 2 months old (juvenile: n = 3), 12 months old (adolescent; n = 3) and 24 months old (adult; n = 3) donkeys. Total RNA was extracted from dissected testicles for miRNA sequencing and analysis. In total, 586 miRNAs, including 451 known miRNAs and 135 novel miRNAs, were identified. Among identified miRNAs, 315 displayed age-dependent expression differences. The levels of miRNA expression in the juvenile group were significantly higher than in the adolescent or adult groups. The MiR-483 exhibited the maximum fold change between juvenile and adolescent groups. Several screened genes, including SLC45A4 and TFCP2L1, have been linked to male reproductive pathways in donkeys. In addition, miR-744 was predicted to regulate SPIN2B, a gene implicated in spermatocyte cell cycle progression and genomic integrity of spermatozoa. These results contribute to our comprehension of microRNA regulation during testicular development and spermatogenesis in Dezhou donkeys. The identified microRNAs and their target genes have the potential to serve as biomarkers for evaluating the reproductive capacity of stud donkeys.

Administration of melatonin prior to modified synchronization protocol improves the productive and reproductive efficiency of Chinese crossbred buffaloes in low breeding season.

Introduction: Melatonin is a neurohormone involving various biological processes, including restoration of cyclicity in animals with seasonal breeding patterns. The use of melatonin in different forms has gained broader acceptance in different species, particularly in summer anestrous buffaloes. Objectives: The objective of the current study was to evaluate the melatonin effect on the reproductive and productive performance of crossbred buffaloes during the low breeding season. Methods: Sixty-five cyclic and reproductively sound crossbred buffaloes were randomly allocated to three groups: the G1 (n = 20) served as the control group and received no single melatonin, G2 received melatonin (n = 22; 18 mg/50 kg, body weight) once prior to synchronization and G3 group was administered multiple melatonin injections (n = 23; 6 mg/50 kg body weight) for three consecutive days before the start of the synchronization protocol. The reproductive performance, milk yield traits, and serum immunoglobulin M (IgM) and melatonin levels were evaluated in treated and untreated crossbred buffaloes. Results: The results revealed that a single dose of melatonin administration has (p < 0.05) improved estrus response, ovulation occurrence and follicular growth in crossbred buffaloes compared to control groups. Higher pregnancy rates were observed in both melatonin-treated buffalo groups compared to the control. Following the administration of melatonin, serum IgM level increased in G2 and G3; however, an increment in melatonin level (p < 0.05) was detected in the G2 group only as compared to the control group subsequent day of melatonin administration. The milk compositions were not affected by melatonin administration except for milk urea nitrogen and somatic cell count (SCC). The melatonin administration (p < 0.05) decreased the somatic cell count in buffalo milk compared to untreated. Conclusion: In conclusion, single or multiple doses of melatonin before initiating the synchronization protocol improved the ovulation, ovulatory follicle diameter and pregnancy rates in crossbred buffaloes during the low breeding season. Moreover, the administration of melatonin enhanced the IgM values along milk traits in terms of milk protein, MUN and somatic cell count in treated buffaloes.

Administration of GnRH at day 20 of AI and embryonic losses in synchronized crossbred buffalo.

Introduction: Following the application of different artificial insemination and synchronization protocols, the synchronized buffaloes had a higher incidence of early embryonic or fetal death, thus impairing reproductive performance. Therefore, there is a need to devise a synchronizing program that can improve conception, allow early pregnancy diagnosis, reduce early embryonic losses, and provide an early clue for pregnancy diagnosis and establishment. The present study aimed (1) to determine the effect of administration of different GnRH doses at day 20 of artificial insemination (AI) on the reproductive performance of buffaloes and (2) to observe the influence of synchronization protocol, parity, age, milk production, and body condition score (BCS) on early embryonic loss and progesterone (P4) levels in crossbred buffaloes. Methods: Crossbred buffaloes (n = 101) were synchronized using the GPGMH protocol. At day 20 of AI, the synchronized buffaloes were randomly divided into four groups and administrated GnRH doses (0, 100, 150, and 200 µg). The buffaloes were monitored for ovarian dynamics, P4 level, non-return rate, embryonic/fetal losses, and pregnancy rates. The previously synchronized buffaloes were also classified for synchronization protocol (with or without GnRH), parity (nulli- or multiparous), milk production (high or low), BCS (low, medium, or good) or age (>3 or < 3 years) groups for observing the embryonic loss and P4 level variations. Results: The results indicated no difference (P > 0.05) in CL size, P4 level, pregnancy rate and embryo/fetal losses across the treatment groups at different observation periods. There was a high (P < 0.05) incidence of early embryonic mortality in aged, multiparous, low BCS and low milk-producing buffaloes treated without GnRH. Conclusion: The data suggest that GnRH 200 µg at day 20 of AI improves embryo survival and pregnancy maintenance in crossbred buffaloes.

Follicular Dynamics during Estrous Cycle of Pubertal, Mature and Postpartum Crossbred (Nili Ravi × Jianghan) Buffaloes.

The follicular dynamics is used as a reliable indicator for reproductive management in livestock. However, the follicular dynamics (follicle wave emergence, estrus cycle length, diameter of dominant follicle, follicular growth and atretic phases) during the estrous cycle of crossbred (Nili Ravi-Jianghan) buffalo is still unexplored. Therefore, the present study aimed to observe the follicular dynamics in estrous cycle of crossbred buffaloes at different physiological stages (pubertal; n = 28, sexual mature; n = 22 and postpartum; n = 18). In the present study, the follicular dynamics were ultrasonically examined at 12 h intervals throughout an estrous cycle during the breeding season. The results indicate that about 86.76% (59/68) crossbred buffaloes, irrespective of physiological stage, exhibited two follicular waves in estrous cycle with an average estrus cycle length was 20.7 ± 0.4 days. The estrus cycle length was significantly shorter (p < 0.05) in pubertal buffaloes (19.4 ± 0.4 days) compared with sexual mature (21.5 ± 0.3 days) and postpartum (21.9 ± 0.4 days) buffaloes. The first follicular wave emerged on same day during one- (pubertal vs. postpartum), two- (pubertal vs. mature vs. postpartum) or three-wave (mature vs. postpartum) estrous cycle buffaloes. The maximum diameter of dominant follicle (DF) in pubertal, sexually mature and postpartum crossbred buffaloes was 9.6 ± 2.0 mm, 10.6 ± 0.5 mm and 12.6 ± 0.7 mm with growth rate of 1.08 ± 0.04 mm/day, 0.92 ± 0.04 mm/day, and 0.9 ± 0.07 mm/day, respectively. In conclusion, similar to other buffalo breeds, Nili Ravi-Jianghan crossbred buffaloes showed the two-wave follicular pattern dominantly with an average duration of ~20 days estrous cycle. The observed follicular dynamics can be used as a reliable indicator for synchronization and fixed-time artificial insemination (FTAI) programs to improve the fertility of crossbred buffaloes.

Effect of Different Synchronization Regimens on Reproductive Variables of Crossbred (Swamp × Riverine) Nulliparous and Multiparous Buffaloes during Peak and Low Breeding Seasons.

The present study was conducted to examine the effect of conventional the Ovsynch protocol (OVS) and a modified Ovsynch synchronization (GPGMH) protocol on the follicular dynamics, estrus, ovulation, and pregnancy in nulliparous and multiparous crossbred (swamp × riverine) buffaloes during different seasons. GPGMH or OVS protocols were used to synchronize nulliparous (n = 128; GPGMH = 94, OVS = 34) and multiparous (n = 154; GPGMH = 122, OVS = 32) buffaloes during the peak (n = 186; GPGMH = 143, OVS = 43) and low breeding (n = 96; GPGMH = 73, OVS = 23) seasons. Buffaloes were monitored for follicular dynamics, estrus response, ovulation, and pregnancy rates. The results showed that protocol, parity, and season had significant effects on estrus, ovulation, and pregnancy variables, and interactions among parity and protocol, season and protocol, and season and parity were observed for few of reproductive indices in the crossbred buffaloes. There were no significant (p > 0.05) interaction for protocol, parity and season. In multiparous buffaloes, the application of the GPGMH protocol significantly (p < 0.05) increased the interaction to the interval to estrus onset after the second GnRH, estrus response, ovulation rate, and pregnancy rate, and lowered (p < 0.05) the silent estrus when compared with the conventional OVS protocol. During the peak breeding season, the application of the GPGMH protocol significantly (p < 0.05) improved the interaction to the estrus response, ovulation rate, and pregnancy rate, while it lowered (p < 0.05) the silent estrus incidence when compared to the conventional OVS protocol. In conclusion, the GPGMH protocol, in comparison to the OVS protocol, improves the follicular dynamics, estrus response, ovulation, and pregnancy rates in crossbred multiparous buffaloes during the peak breeding seasons.

Dietary Supplementation of Capsaicin Enhances Productive and Reproductive Efficiency of Chinese Crossbred Buffaloes in Low Breeding Season.

The present study investigated the role of dietary capsaicin (CPS) supplementation on milk yield (liters/head) and milk composition (total solids, lactose, albumin, protein, fat, milk urea nitrogen (MUN), somatic cell count (SCC) and somatic cell score (SCS), serum metabolites (lipoprotein esterase (LPL) and aspartate aminotransferase (AST)), and reproductive physiology (follicular development, estrus response, ovulation and pregnancy) following synchronization during the low breeding season. One hundred (n = 100) crossbred buffaloes were randomly assigned to four dietary groups consisting of CPS supplementation dosages (0, 2, 4 or 6 mg/kg of total mixed ration; TMR) as CPS-0 (n = 26), CPS-2 (n = 22), CPS-4 (n = 25) and CPS-6 (n = 27), respectively, in a 30-day feed trial. The results revealed that the CPS-4 group of buffaloes had a better estrus rate (72%) along with improved (p < 0.05) ovulatory follicle diameter (13.8 mm), ovulation rate (68%) and pregnancy rate (48%) compared to other treatment groups. Milk yield improved (p < 0.05) in CPS-4 supplemented buffaloes after day 20 of the trial, comparatively. There was a significant effect (p < 0.05) of milk sampling day (day 30) on total milk solids, lactose, milk protein and MUN levels, whereas lactose, MUN, SCC and SCS were influenced by supplementation dosage (CPS-4). Glucose levels were affected in buffaloes by sampling time (artificial insemination (AI) and 50-day post-AI) and CPS-dose (CPS-4 and CPS-6), respectively. LPL level changed in CPS-2 and CPS-4 groups at AI time and 50 days after AI. In addition, the AST level was different in CPS-4 at AI time and 50 days after AI. Therefore, our data suggest that a medium dose (~4 mg/kg of TMR) of CPS provided a better response in the form of milk yield, milk composition, serum metabolites and reproductive performance in crossbred buffaloes during the low breeding season.

GPGMH, a New Fixed Timed-AI Synchronization Regimen for Swamp and River Crossbred Buffaloes (Bubalus bubalis).

The crossbreeding of Swamp and River type buffalo breeds is practiced for the improvement of milk yield and reproductive performance in swamp buffalo herds. This study aimed to modify the Ovsynch synchronization protocol (GPG) and improve the fixed-timed artificial insemination (FTAI) for better reproductive performance of crossbred buffaloes. Comparison of four conventional synchronization protocols [pregnant mare gonadotropin-prostaglandin F2α-gonadotropin-releasing hormone (PmPG), gonadotropin-releasing hormone-prostaglandin F2α-gonadotropin-releasing hormone (GPG), prostaglandin F2α-gonadotropin-releasing hormone-prostaglandin F2α-estradiol benzoate (PGPE), and progesterone-pregnant mare gonadotropin-prostaglandin F2α-gonadotropin-releasing hormone (P4PmPG)] in crossbred buffaloes showed that the GPG protocol treated buffaloes displayed higher (P < 0.05) estrus response with an increasing tendency in ovulation (84.6%) and pregnancy rates (30.8%) than PmPG, PGPE, and P4PmPG treated buffaloes. Buffaloes treated with a dose of 0.4 (mg/kg) mifepristone combined with GPG, exhibited higher (P < 0.05) estrous response (82.4%), ovulation (94.1%), and pregnancy (47.1%) rates compared with other doses (0, 0.3, or 0.5 mg/kg) groups. Injection of mifepristone along second GnRH injection in buffaloes improved (P < 0.05) pregnancy rate (35.3%) when compared to before or after the second GnRH of GPG protocol. Single AI after 24 h of mifepristone or second GnRH injection seems the best time to enhance the pregnancy rates in buffaloes compared to double or other single AI times in the modified GPGMH protocol. In comparison, GPGMH reduced the follicular cyst incidence (P < 0.05) with increasing ovulation (P > 0.05) and pregnancy rates (P > 0.05) than the P4GPG and GPG protocols in crossbred buffaloes. The current study supported that new synchronization protocol (modified of GPG protocol; GPGMH) by the inclusion of mifepristone (with a dose of 0.4 mg/kg along second GnRH), AI after 24 h of mifepristone or second GnRH, and human chorionic gonadotropin (hCG at day 5 of AI) enhance the ovulation and pregnancy rates in crossbred buffaloes.

Protective effect of coenzyme Q10 against bisphenol-A-induced toxicity in the rat testes.

The present study was conducted to investigate the antioxidant, histomorphometric, histochemical, immunohistochemical, biochemical, and cytological effects of coenzyme Q10 (CoQ10) against bisphenol-A (BPA)-induced testicular toxicity in rats. A total of 40 adult male Wistar rats were divided into five equal groups. The control group remained untreated. The vehicle control group was administered corn oil (2 ml/kg/day), the BPA group was given BPA (100 mg/kg/day), the CoQ10 group was supplemented with CoQ10 (10 mg/kg/day), and the rats in the CoQ10-BPA group received CoQ10 (10 mg/kg/day) followed by BPA (100 mg/kg/day) 1 h later. The treatments were administered by oral gavage for 14 days. Results showed that the seminiferous tubule diameters (STDs) and seminiferous epithelium heights (SEHs) at stages VII-VIII and XII-XIV, number of undifferentiated embryonic cell transcription factor-1 (UTF-1) positive cells per tubule, UTF-1 positive tubules (%), plasma glutathione (GSH), and serum superoxide dismutase activities, testicular GSH activity and sperm viability (%) decreased whereas the number of terminal dUTP nick end labeling (TUNEL) positive cells per tubule, TUNEL positive tubules (%), testicular and serum malondialdehyde (MDA) levels, and the rate of mid-piece sperm abnormality increased in the BPA administered group. However, while the STDs at stages VII-VIII and XII-XIV, SEHs at stages VII-VIII, plasma GSH, and serum SOD activities increased, serum MDA level decreased in the CoQ10-BPA group. In conclusion, these results suggest a protective effect of CoQ10 against BPA-induced testicular toxicity in rats.

Effects of long-term release GnRH agonist "deslorelin" on testicular HSP expression, accessory sex glands and testicular functions in adult male rats.

The objective of the present was to determine the effect of long-term release GnRH agonists "deslorelin" on suppression and restoration of testicular and accessory sex glands functions, and expression of HSP in testes of adult male rats. A group of twenty-eight male rats and fifty-six female rats were kept for eleven months. The male rats were subdivided into treatment (n = 18; deslorelin, an analogue of GnRH, 4.7 mg, S.C; six months) and control (n = 10; untreated), and the adult female rats were introduced with either treatment or control male rats at the 2nd, 6th and 11th months post implant insertion. At 6th month of deslorelin implants insertion, six male rats from treatment and five rats from control group were sacrificed. The remaining (twelve treatment and five control) male rats were sacrificed at 11 months. The testicular dimension were measured monthly in both treatment and control rats. The blood samples were collected for testosterone and HSP70 antibody, whereas, the testes and accessory glands were isolated for histological examination at each sacrificial time. The results showed that testicular dimension were significantly lesser in treatment group until 9 months post treatment. HSP70 protein expression was negligible at 6 months in treatment group but its intensity increased in spermatids 11 months of treatment similar to control group. Significantly lower testosterone concentrations with poor semen quality, and smaller litter size were observed in treatment group. The histological picture of accessory sex glands and seminiferous tubules shown a variable integrity in treatment group than control at 6 months implant insertion. In conclusion, the subcutaneous application of 4.7 mg of the GnRH-analogue deslorelin represents a practicable, like in the female rats, method to suppress testicular, accessory sex glands functions, testicular HSP expression and fertility in male rats. Moreover, the suppressive effects of deslorelin, continued until 11th months after removal of the implant.

Seasonal variations in sperm acrosome reaction, osmotic tolerance and serum testosterone concentrations in rams.

The study was conducted to evaluate the seasonal dynamics in the sperm acrosome reaction (AR), osmotic tolerance and serum testosterone concentration in rams. Blood and semen samples were collected from six mature rams during the winter, spring, summer and autumn seasons. The AR in sperm samples was induced by using lysophosphatidylcholine (LPC) and calcium ionophore (CI). The AR was monitored at various incubation times (15, 30, 60, 120 and 180 min) post-induction. To determine the osmotic tolerance through hypo-osmotic swelling test (HOST) the sperm were exposed to different osmotic conditions (50, 100, 290, 500 and 1000 mOsm/L). The serum testosterone concentration was assessed using a solid phase enzyme-linked immunosorbent assay (ELISA). The proportion of acrosome reacted sperm was greater (P < 0.05) during the winter season in the LPC and control groups, whereas, it was greater (P < 0.05) in two different seasons (winter and summer) at all incubation time points (15-180 min) in the CI group. The live HOST +ve, total HOST +ve and total live sperm were least (P < 0.05) during the winter season. The greatest membrane integrity (P < 0.05) was in the autumn season. The peak (P < 0.05) serum testosterone concentration was during the summer season. In conclusion, the osmotic tolerance and sensitivity of ram sperm to undergo the AR in response to LPC and CI vary with season. The increased testosterone concentration in the summer season might be the reason for a greater membrane integrity during the subsequent autumn season in Kivircik ram sperm.

Dietary quercetin maintains the semen quality in rabbits under summer heat stress.

This study focused to determine beneficial impact of feeding quercetin supplemented diet on semen quality in summer heat imposed rabbits. Twelve heat stressed (HS) adult rabbits bucks were either fed with basal diet (HS; n = 06) or quercetin supplemented diet (QU-HS; n = 06) for a period of 56 days. Semen samples were collected and evaluated for volume, osmolality, morphology, concentration, motility, motion kinetics, viability, acrosome integrity, mitochondrial potential, and seminal plasma MDA level. Semen volume, concentration, motility and sperm kinetics parameters were affected by diet supplementation. Diet affected the sperm mitochondrial potential and day of treatment affected the viable sperm percentage. There was an effect of diet, day of treatment and diet by day interaction on acrosome reaction rate. Sperm head abnormalities were influenced by diet provision, sperm mid-piece abnormalities were affected by diet and day of treatment, whereas, the effect of diet and diet by day of treatment interaction were observed for total sperm abnormalities. There was an effect of diet and diet by day interaction for seminal plasma MDA level. In conclusions, quercetin reduces the damaging effects of HS and maintains the semen quality by lowering the oxidative stress in rabbits.

Quercetin supplemented diet improves follicular development, oocyte quality, and reduces ovarian apoptosis in rabbits during summer heat stress.

The present study was designed to test the modulatory effect of dietary quercetin on follicle population, apoptosis, in vitro maturation rate and quality of oocytes in heat stressed female rabbits. A total of thirty-four New Zealand White heat stress (HS) exposed female rabbits were either fed with quercetin supplemented diet (QU-HS) or non-supplemented (HS) diet. Firstly, laparotomy was performed for oocyte retrieval and then, oocyte grading and COCs dimensional assessments were conducted. The A and B-grade oocytes were submitted for in vitro maturation. Thereafter, the ovaries were collected from rabbits and were processed for follicular population estimation and granulosa cells apoptosis. The results showed that follicle number, retrieved oocytes and A-grade oocytes were higher in QU-HS, comparatively. A significant difference was observed in A-grade oocytes dimensions between QU-HS and HS treatment groups. The oocyte maturation rate was same across the groups. The quercetin supplementation significantly improved primordial and antral stage follicles. A greater number of apoptotic cells were observed in primary and antral follicles in the HS group. In conclusion, the quercetin provision improves the follicular development, minimize granulosa cells apoptosis, and maintain the oocyte competence in HS rabbits.

Royal jelly supplementation in semen extender enhances post-thaw quality and fertility of Nili-Ravi buffalo bull sperm.

Two experiments were conducted to evaluate the effect of royal jelly (RJ) on post-thaw sperm quality, in vitro and in vivo fertility rate of cryopreserved buffalo bull sperm. The semen was collected from three mature regular donor buffalo bulls, ejaculates were pooled and semen evaluated initially. In Experiment 1, the ejaculates were extended in tris-citric acid diluter supplemented with different RJ concentrations (0, 0.05, 0.1, 0.2, 0.3 or 0.4%). The diluted semen was cooled to 4°C, packaged into 0.5 mL straws and frozen using standard procedure. The straws were thawed and assessed for sperm progressive motility, viability, plasma membrane, acrosome, and chromatin integrity. The results indicated that sperm progressive motility was significantly greater (P<0.05) in 0.05, 0.1, 0.2 and 0.3% RJ than 0.4% RJ supplemented and control groups. The sperm viability, plasma membrane and acrosome integrity were significantly improved (P<0.05) in 0.1% RJ supplemented group the compared to other treatment groups. In Experiment 2, cryopreserved sperm with 0.1% RJ supplementation and control (without RJ supplementation) were used to observe the in vitro fertilizing potential and in vivo fertility. In vitro fertilization method was applied to assess the cleavage rate; whereas, AI was performed in buffalo during in vivo fertility trial. The buffaloes were inseminated 12h after standing estrus and pregnancy diagnosis was performed through ultrasonography. The results revealed that the cleavage rate was higher (P<0.05) in 0.1% RJ as compared to control group. However, the pregnancy rate was similar (P>0.05) between 0.1% RJ supplemented and control groups. It is concluded that supplementation of RJ in freezing extender can improve the cryosurvival rate and in vitro fertilizing capacity of buffalo bull sperm.

Protective effect of cholesterol-loaded cyclodextrin pretreatment against hydrogen peroxide induced oxidative damage in ram sperm.

Three experiments were conducted to determine the protective effect of cholesterol-loaded cyclodextrin (CLC) against hydrogen peroxide (H2O2) or cryo-induced damage in ram sperm. In Experiment 1, the fresh ejaculates were either treated with CLC or remained untreated. Both CLC treated and untreated samples were then incubated with 0, 250 or 500 µM H2O2 at 35°C for 12 h. After incubation period of 12 h, the motility, viability and membrane integrity remained higher in CLC treated sperm even in the presence of 250 or 500 µM H2O2. The H2O2 treatment affected all the sperm parameters adversely (P<0.05). However, compared to CLC untreated counterpart, the motility, viability and membrane integrity remained higher (P<0.05) in treated sperm, even in the presence of 250 or 500 µM H2O2 during 12 h of incubation. In Experiment 2, semen was cryopreserved in the presence or absence of CLC. The post-thaw results revealed that CLC treated sperm has higher (P<0.05) motility, viability and membrane integrity compared to the control. In Experiment 3, lipid peroxidation levels were assessed by determining malondialdehyde (MDA) concentrations during the H2O2-induced oxidative stress in CLC treated and untreated sperm. However, no difference (P>0.05) in MDA level was observed among the groups at any stage of incubation. In conclusion, the CLC incorporation in ram sperm membrane may protects it against H2O2 or cryo-induced oxidative damage. The cryoprotective influence of CLC on ram sperm might be resulted from, at least partly, its antioxidative property.

Comparison of two different cryopreservation protocols for freezing goat semen.

In this study, two different semen cryopreservation protocols were compared to freeze goat semen. The ejaculates (n=12) were collected by using electro-ejaculator from six mature bucks (two ejaculates per each buck). Each ejaculate was divided into two groups as Protocol 1 (P1) and Protocol 2 (P2). In P1, semen was diluted directly in an extender containing 15% egg yolk, 300mM Tris, 28mM glucose, 95mM citric acid 5% glycerol to a concentration of 200×10(6)sperm/mL. In P2, after the removal of seminal plasma by centrifugation, the semen sample was diluted with the first portion of milk extender consist of 100mg/mL skimmed milk powder and 27.75mM glucose (without glycerol) to a concentration of 400×10(6)sperm/mL. The second portion of the milk extender containing 14% glycerol was added to semen gradually in order to achieve sperm concentration 200×10(6)sperm/mL and 7% glycerol level in the final volume. Extended semen was loaded in 0.25mL straws, held for 2h at 4°C, frozen in nitrogen vapor and stored in liquid nitrogen. Post-thaw motility and live sperm rate (mean±SEM) were significantly lower (P<0.05) in P1 as compared to P2 (47.50±1.23% vs. 55.63±1.72%; 80.04±1.29% vs. 84.04±1.08%, respectively). However, live intact, total intact, abnormal, reacted acrosome and DNA damaged sperm rates were similar (P>0.05) in both protocols. It was concluded that both protocols used in this study provided reasonable post-thaw parameters; however, P2 yielded better motility and live sperm rate compared to P1.

Exploring arising moments and mindfulness in occupational therapists working in diverse clinical practice areas.

An online survey was conducted with 72 Canadian occupational therapists to (1) explore if and how occupational therapists were aware of "arising moments" in clinical practice, which are moments that give rise to emotions, sensations, and thoughts, and (2) to examine therapists' responses to questions related to mindfulness. Participants described arising moments through an open-ended survey question. Framework analysis was used to develop key themes and subthemes. A modified Philadelphia Mindfulness Awareness (PHLMS) subscale with two additional survey questions "understanding of living in the moment and being present, and awareness of mindfulness" (sum = PAM) were used as indicators of mindfulness. There were significant differences in the means of PHLMS mindfulness among therapists working in diverse practice areas (F = 3.63, p = .009). Posthoc analysis revealed that therapists working in mental health had higher mean PHLMS-mindfulness scores than in all other groups. There were no significant differences in PAM mindfulness among the practice areas (ANOVA, F = 2.15, p = .08). However, posthoc tests showed that the participants in the mental health practice area had a significant difference with one group, general physical health. Findings have implications for practice in occupational therapy and education about mindfulness in occupational therapy.

Relationship of age to body weight, scrotal circumference, testicular ultrasonograms, and semen quality in Sahiwal bulls.

The objective of the present study was to determine the relationship of age to body weight (BW), scrotal circumference (SC), number of pixels of testicular ultrasonograms (NP), and semen quality in Sahiwal bulls. The study was based on 128 Sahiwal bulls of different age groups (from 0 to >100 months of age). Bulls were evaluated for SC, BW, and NP. Semen was evaluated once a week for five consecutive weeks from regularly collected donor bulls (n = 86) ranging in age from 25-30 to >100 months. Ejaculate volume, sperm motility, sperm concentration, sperm morphology, percent live sperms, sperm plasma membrane integrity, and normal acrosome were compared among different age groups. Mean SC and BW increased (P < 0.05) in a curvilinear manner from birth to >100 months of age. Mean NP of testicular ultrasonograms increased (P < 0.05) from 0 to 24 months and then plateaued until >100 months of age. Body weight, SC, and NP were positively correlated with age from birth until >100 months (r = 0.91, 0.87, and 0.40, respectively). Ejaculate volume (5.7 ± 0.2 vs. 4.6 ± 0.09 ml) and sperm concentration (1,281.6 ± 17.7 vs. 1,115.8 ± 55.9 × 10(6)/ml) increased (P < 0.05) in mature bulls compared to younger ones. However, motility (68.6 ± 0.3%), plasma membrane integrity (50.8 ± 1.0%), and normal acrosome (74.8 ± 0.5%) remained insignificant due to age. In six of eight age groups studied, morphological abnormalities were well within the range (18.1 ± 0.3%). In conclusion, the BW, SC, and NP of testicular ultrasonograms, ejaculate volume, and concentration increased with age. Moreover, semen quality is fairly independent of age except volume and concentration in Sahiwal bulls.

A long-term, open-label study to confirm the safety of topical diclofenac solution containing dimethyl sulfoxide in the treatment of the osteoarthritic knee.

To assess the long-term safety of a topical solution of diclofenac sodium in a vehicle containing dimethyl sulfoxide (TDiclo), subjects with radiologically confirmed, symptomatic osteoarthritis of the knee(s) applied the clinical dose of TDiclo (40 drops, four times daily) to each painful knee for up to 52 weeks. Safety assessment included adverse events, skin irritation scores of the treated knee(s), ocular examinations, and routine laboratory tests. There were 793 subjects (mean age, 62.5 years) who applied TDiclo to one or both (72%) knees for an average of 204 days, including 463 subjects for 6 months and 144 for 1 year. The most frequent adverse events were at the application site with no increase in incidence with prolonged exposure: dry skin (25.3%), contact dermatitis without vesicles (13.0%) or with vesicles (9.5%). Skin irritation score was 0 (normal) in 61.0% of subjects, 0.5 (dryness or flaking) in 23.9%, 1 or 2 (erythema without or with induration) in 6.9%, and 3 or 4 (erythema with induration and vesicles/bullae) in 8.2%. Subject dropouts included 114 (14.4%) with an application site skin adverse event. Individual subject laboratory test shift to abnormal occurred for hemoglobin (3.2%), aspartate aminotransferase (6.4%), alanine aminotransferase (7.3%), and creatinine (4.2%), but few shifts (less than 0.3% per variable) were clinically significant. No increased risk of cardiovascular or cataract events was noted. This long-term study of TDiclo revealed a safety profile comparable to that shown in multiple, shorter, well-controlled, double-blind trials with the predominant adverse effect noted being an application site reaction.

Efficacy and safety of topical diclofenac containing dimethyl sulfoxide (DMSO) compared with those of topical placebo, DMSO vehicle and oral diclofenac for knee osteoarthritis.

While topical non-steroidal anti-inflammatory drugs are considered safe, their long-term efficacy for osteoarthritis has been suspect. We conducted a 12-week, double-blind, double-dummy, randomized controlled trial of topical diclofenac (TDiclo) in a vehicle solution containing dimethyl sulfoxide (DMSO) in 775 subjects with radiologically confirmed, symptomatic primary osteoarthritis of the knee. This 5-arm study compared TDiclo with a placebo solution, the DMSO vehicle, oral diclofenac (ODiclo) and the combination of TDiclo+ODiclo for relieving the signs and symptoms of knee osteoarthritis. Subjects applied study solution, 40 drops four times daily, and took one study tablet daily for 12 weeks. Co-primary efficacy variables were WOMAC pain and physical function and a patient overall health assessment. Secondary variables were WOMAC stiffness and patient global assessment (PGA) of the knee osteoarthritis. TDiclo was superior to placebo for pain (-6.0 vs. -4.7, P=0.015), physical function (-15.8 vs. -12.3, P=0.034), overall health (-0.95 vs. -0.37, P<0.0001), and PGA (-1.36 vs. -1.01, P=0.016), and was superior to DMSO vehicle for all efficacy variables. No significant difference was observed between DMSO vehicle and placebo or between TDiclo and ODiclo. The commonest adverse event associated with TDiclo was dry skin (18.2%). Fewer digestive system and laboratory abnormalities were observed with TDiclo than with ODiclo. Addition of TDiclo to ODiclo did not increase the incidence of systemic adverse events. TDiclo in DMSO vehicle is an effective treatment option for knee osteoarthritis with efficacy similar to, but tolerability better than ODiclo. DMSO vehicle was no more efficacious than placebo.