scholarly journals Neutrophil recruitment and phagocytosis of boar spermatozoa after artificial insemination of sows, and the effects of inseminate volume, sperm dose and specific additives in the extender

Reproduction ◽  
2003 ◽  
pp. 357-367 ◽  
Author(s):  
A Matthijs ◽  
B Engel ◽  
H Woelders

In this study the recruitment of leucocytes and phagocytosis of spermatozoa after artificial insemination of multiparous sows was investigated. In Expt 1, groups of sows received either no inseminate (n = 6) or inseminates with various concentrations of spermatozoa and seminal plasma or different inseminate volumes (n = 9 per group). In Expt 2, groups of sows received inseminates containing no addition, caffeine + CaCl(2), or excess EDTA (n = 6 per group). Leucocytes and spermatozoa were counted in the collected backflow from the vulva, and in the PBS flushings of the genital tract of sows killed at 4 h after insemination. Tissue homogenates were checked for remaining spermatozoa. Leucocyte recruitment did not depend on the presence of seminal plasma or spermatozoa. In the control groups about 43% of the inseminated spermatozoa were found in the backflow and 5% in the genital tract. Many spermatozoa could be recognized inside polymorphonuclear leucocytes. With an inseminate volume of 20 ml instead of 80 ml, fewer spermatozoa were found in the backflow and more (non-phagocytosed) spermatozoa were recovered in the uterus (P < or = 0.05). With a sperm dose of 0.24 x 10(9) instead of 2.4 x 10(9), a higher percentage of the inseminated spermatozoa was recovered in the oviducts (P < or = 0.05). The use of caffeine + CaCl(2) resulted in lower recruitment of leucocytes (P < or = 0.05) and a higher number of non-phagocytosed spermatozoa in the uterus (P < or = 0.01) compared with controls. The numbers of spermatozoa in the oviducts were not different. Insemination with excess EDTA had no positive effects on the number of spermatozoa in the genital tract.

2004 ◽  
Vol 16 (2) ◽  
pp. 255
Author(s):  
H. Funahashi

Fertility of boar spermatozoa as determined following artificial insemination seems to be maintained during liquid preservation at 10–15°C for several days, although prolonged liquid preservations reduce the pregnancy rate rapidly. However, it is not clear if spermatozoa can penetrate into oocytes in an IVF system even after a prolonged liquid preservation. Oxidative stress could also be one of the possible detrimental factors in liquid preservation of spermatozoa. In the present study, fertility of liquid-preserved spermatozoa was examined using an IVM-IVF system. Whether cysteine can improve the fertility was also determined. Spermatozoa (from four Berkshires) was resuspended at 1×108 cells mL−1 in Modena solution containing 15% (v/v) boar seminal plasma and 0 or 5mM cysteine after washing 3 times. Sperm suspensions (1mL) were then preserved at 10°C for 22 days following a program for cooling down (to 15°C for 4h, keeping at 15°C for 12h and then to 10°C for 6h). At Days 1, 8, 15 and 22 after the start of preservation, spermatozoa (5×105 cells mL−1) were co-cultured with IVM oocytes in an IVM/IVF system (Funahashi et al., 1997 Biol Reprod 57, 49–53). Viability and functional status of spermatozoa were also examined at Days 8 and 15 of preservation by using LIVE/DEAD sperm viability kit and CTC fluorescence assay. Data (mean±SEM) from 4–6 replicates were analyzed by ANOVA and Fisher’s protected LSD test. When spermatozoa that had been preserved without cysteine (Cys−) were used, penetration rates were not different (P&gt;0.05) from those with cysteine (Cys+) at Day 8 of preservation (91.4±3.4% in Cys− and 99.3±0.7% in Cys+), but lower (P&lt;0.02) at Days 15 and 22 (72.6±13.6% and 33.8±8.4% in Cys−; 94.8±2.1% and 71.1±10.8% in Cys+, respectively). Both viability and proportion of uncapacitated live cells were higher (P&lt;0.05) in Cys+ than Cys− at Days 8 and 15. These results demonstrate that boar spermatozoa can penetrate into oocytes in vitro even after a liquid preservation at 10°C for 22 days and that cysteine can improve the viability and penetrability in vitro of spermatozoa during liquid preservation. Supported by the Ito Foundation.


2011 ◽  
Vol 48 (4) ◽  
pp. 164-168 ◽  
Author(s):  
Tetsuji OKAZAKI ◽  
Teiichi AKIYOSHI ◽  
Masakazu KAN ◽  
Hisanori TESHIMA ◽  
Masayuki SHIMADA

2003 ◽  
Vol 83 (2) ◽  
pp. 221-227 ◽  
Author(s):  
A. Morrier ◽  
F. Castonguay ◽  
J. L. Bailey

Seminal plasma aids sperm transport and contains factors beneficial for sperm function. In artificial insemination, however, diluting the semen reduces the concentration of seminal plasma. To test the hypothesis that supplemental seminal plasma in extended ram semen improves conservation at 5°C, we added various concentrations of seminal plasma to semen during storage, and investigated subsequent sperm function in vitro. Semen was divided into three aliquots, extended in a commercial diluent (Triladyl) supplemented with 0, 10 or 25% (vol:vol) ovine seminal plasma and cooled to 5°C. After 8 and 24 h at 5°C, sperm were suspended in a modified synthetic oviduct fluid (SOF-m) at 39°C to mimic the female genital tract at insemination. Sperm aliquots were assessed for motility and chlortetracycline fluorescence after 0, 4 and 8 h in the SOFm. No significant differences were observed due to seminal plasma supplementation during conservation at 5°C or incubation in SOF-m at 39°C. However, decreased sperm motility and fewer non-capacitated sperm were observed concomitant with an augmentation of capacitated and acrosome-reacted cells during incubation in SOF-m. Therefore, the hypothesis that diluent supplementation with homologous seminal plasma improves ram sperm conservation or subsequent sperm function was not supported. Key words: Ovine, ram, sperm, motility, viability, chlortetracycline fluorescence, artificial insemination, SOF


2021 ◽  
pp. 097275312110000
Author(s):  
Madhava Sai Sivapuram ◽  
Vinod Srivastava ◽  
Navneet Kaur ◽  
Akshay Anand ◽  
Raghuram Nagarathna ◽  
...  

Background: Type 2 diabetes needs a better understanding of etiological factors and management strategies based on lifestyle and constitutional factors, given its high association rate with many cardiovascular, neurological disorders, and COVID-19 infection. Purpose: The present study was undertaken to investigate the effect of Diabetes-specific integrated Yoga lifestyle Protocol (DYP) on glycemic control and lipid profiles of diabetic adults. Along with the DYP intervention, the individuals residing in Chandigarh and Panchkula union territories in the northern part of India were assessed for Ayurveda-based body–mind constitutional type. Ayurveda describes body–mind constitution as “ prakriti,” which has been discussed from two angles, namely physiological and psychological as body and mind are correlated. Methods: Cluster sampling of waitlist control study subjects was used as the sampling method for the study. A total of 1,215 registered subjects (81 diabetic) responded in randomly selected clusters in Chandigarh and Panchkula. Ayurveda physicians did Ayurveda body–mind constitutional assessment called prakriti assessment (physiological body–mind constitution assessment) in 35 participants (23 diabetic, 12 prediabetic) as a part of the study. Results: A group of 50 subjects was randomly selected for yoga intervention out of 81 diabetes mellitus adults, and 31 subjects were enrolled as waitlist controls. A significant decrease in the glycosylated hemoglobin levels from 8.49 ± 1.94% to 7.97 ± 2.20% in the intervention group was noticed. The lipid profiles of the DYP intervention and control groups were monitored. Three-month follow-up results of lipid profile diagnostic tests in intervention and control groups showed a significant difference between the two groups ( P < 0.05). Most diabetic and prediabetic individuals were found to have pitta dosha ( pitta controls all heat, metabolism, and transformation in the mind and body) as dominant constitution type. Conclusion: The study results demonstrated significant positive effects of yoga in diabetic individuals. This study has indicated the evidence for the safety and efficacy of the validated DYP for community-level interventions to prevent maladies like brain damage and stroke.


2017 ◽  
Vol 29 (8) ◽  
pp. 1576 ◽  
Author(s):  
Rocío Fernández-Gago ◽  
Manuel Álvarez-Rodríguez ◽  
Marta E. Alonso ◽  
J. Ramiro González ◽  
Beatriz Alegre ◽  
...  

Seminal plasma could have positive effects on boar semen after thawing. In the present study we investigated changes in the motility and chromatin structure in spermatozoa over 4 h incubation (37°C) of boar semen thawed in the presence of 0%, 10% or 50% seminal plasma from good-fertility boars. Cryopreserved doses were used from seven males, three of which were identified as susceptible to post-thawing chromatin alterations. Motility was analysed by computer-aided sperm analysis every hour, and data were used in a two-step clustering, yielding three subpopulations of spermatozoa (slow non-linear, fast non-linear, fast linear). Chromatin structure was analysed using a sperm chromatin structure assay and flow cytometry to determine the DNA fragmentation index (%DFI) as a percentage, the standard deviation of the DFI (SD-DFI) and the percentage of high DNA stainability (%HDS), indicating chromatin compaction. Thawing without seminal plasma resulted in a rapid loss of motility, whereas seminal plasma helped maintain motility throughout the incubation period and preserved the subpopulation comprising fast and linear spermatozoa. The incidence of chromatin alterations was very low in samples from non-susceptible males, whereas samples from males susceptible to post-thawing chromatin alterations exhibited marked alterations in %DFI and %HDS. Seminal plasma partly prevented these alterations in samples from susceptible males. Overall, 50% seminal plasma was the most efficient concentration to protect motility and chromatin. Some changes were concomitant with physiological events reported previously (e.g., semen thawed with 50% seminal plasma increased the production of reactive oxygen species and yielded higher fertility after AI). Thawing in the presence of seminal plasma could be particularly useful in the case of samples susceptible to post-thawing chromatin damage.


2021 ◽  
Vol 8 ◽  
Author(s):  
Eduardo G. Aisen ◽  
Wilfredo Huanca López ◽  
Manuel G. Pérez Durand ◽  
Edita Torres Mamani ◽  
Juan C. Villanueva Mori ◽  
...  

The viscous seminal plasma (SP) is currently a major impediment to the handling of ejaculate and the development of some biotechnologies in South American camelids. The vas deferens-collected spermatozoa of alpacas is a useful technique to avoid this problem. On the other hand, SP contains a large protein component that has been implicated in the function of spermatozoa within the female reproductive tract. In this sense, the low fertility achieved using transcervical insemination with frozen-thawed spermatozoa in alpacas could be improved by adding SP. This study aimed to evaluate the effect of the whole SP on some in vitro parameters of alpaca spermatozoa after the freezing-thawing-process and the fertility after artificial insemination. It would contribute to a better understanding of the interaction between thawed sperm cells and SP. Spermatozoa were obtained by surgically diverted vas deferens. The samples were diluted with a Tris-based extender, packaged in straws, and frozen. At thawing, each straw was divided into two post-thawing conditions: with the addition of 10% of PBS (control) or with 10% SP (treatment). The sperm cells were evaluated using dynamic parameters, sperm cell morphology, and morphometry. Fertility was assessed by an artificial insemination trial. All in vitro parameters were analyzed by ANOVA. A heterogeneity test was scheduled for the fertility trial. After the freezing-thawing process, motility and plasma membrane functionality was improved when SP was added. No differences were found for post-thaw viability between the control and treatment samples. The percentage of normal cells was higher with SP at post-thawing, and a decrease of the presence of bent tailed spermatozoa with a droplet in the SP group was observed. The length of the head spermatozoa was 3.4% higher in the samples with PBS compared to those in which SP was added. Females pregnant at day 25 post-insemination were 0/12 (with SP inside the straw) and 1/10 (without SP inside the straw). In conclusion, the presence of 10% SP at post-thawing improves sperm cells' motility, functionality, and morphology, indicating that it would be beneficial to improve the frozen-thawed alpaca's physiology spermatozoa. More fertility trials must be developed to increase this knowledge.


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