Kim, Jin-Hee;Chung, Ee-Yung;Choi, Ki-Ho;Park, Kwan-Ha;Park, Sung-Woo
한국패류학회지
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제26권1호
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pp.33-43
/
2010
The ultrastructure of germ cells during spermatogenesis and some characteristics of sperm morphology in male Mytilus coruscus, which was collected on the coastal waters of Gyeokpo in western Korea, were investigated by transmission electron microscope observations. The morphology of the spermatozoon has a primitive type and is similar to those of other bivalves in that it contains a short midpiece with five mitochondria surrounding the centrioles. The morphologies of the sperm nucleus type and the acrosome shape of this species have an oval and modified cone shape, respectively. In particular, the axial rod is observed between the nucleus and acrosome of the sperm. The spermatozoon is approximately $45-50{\mu}m$ in length including a sperm nucleus (about $1.46{\mu}m$ in length), an acrosome (about $3.94{\mu}m$ in length) and tail flagellum (approximately $40-45{\mu}m$). The axoneme of the sperm tail flagellum consists of nine pairs of microtubules at the periphery and a pair at the center. The axoneme of the sperm tail shows a 9+2 structure. Some special charateristics of sperm morphology of this species in the genus Mytilus are (1) acrosomal morphology, (2) the number of mitochondria in the midpiece of the sperm, and (3) the existence of a satellite. The axial rod appears in the acrosome and sperm nucleus as one of the characteristics seen in several species of the subclass Pteriomorphia, unlikely the subclass Heterodonta containing axial filament instead of the axial rod. The number of mitochondria in the midpiece of the sperm of this species in the family Mytilidae are five, as one of common characteristics appeared in most species in the family Mytilidae. Most of Mytilus species contain a satellite body which is attached to the proximal centriole in the middle piece of the sperm, as one of common characteristics of sperm morphology in the family Mytilidae.
This study was carried out determine whether Unopette can be used for the observation of sperm morphology and sperm Concentration. Rabbit sperm and frozen-thawed bovine sperm were observed with phase contrast microscope after dilution with Unopette acooriding to duration of preservation at 3~5$^{\circ}C$. Sperm using Unopette showed high normal sperm(%) than sperm using hematoxylin-eosin until 48 hours. Sperm using Unopette revealed no difference in sperm concentration until 24 hours, as compared with control sperm. As a result, Unopette was assessed as appropriate solution for preservation in terms of morphological observation and sperm concentration.
Park, Yong-Seog;Park, Sol;Ko, Duck Sung;Park, Dong Wook;Seo, Ju Tae;Yang, Kwang Moon
Clinical and Experimental Reproductive Medicine
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제41권3호
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pp.132-136
/
2014
Objective: The presence of sperm-head vacuoles has been suspected to be deleterious to the outcomes of assisted reproductive technology (ART). It is difficult to accurately distinguish morphologically abnormal sperm with vacuoles under a light microscope. This study was performed to analyze the result of the observation of sperm-head vacuoles using Papanicolaou staining under a light microscope and whether the male partner's age affects these vacuoles. Methods: Sperm morphology with vacuoles was evaluated using Papanicolaou staining and observed under a light microscope ($400{\times)$) in 980 men. The normal morphology was divided into three categories (group A, <4% of normal morphology; group B, 4%-14% of normal morphology; and group C, >14% of normal morphology). The criteria for the sperm-head vacuoles were those given in the World Health Organization manual. For the analysis of the age factor, the participants were divided into the following groups: 26-30 years, 31-35 years, 36-40 years, 41-45 years, and 46-50 years. Results: The percentage of sperm-head vacuoles increased with normal sperm morphology (group A vs. groups B, C) (p<0.05). In the case of the age factor, a statistically significant difference was not observed across any of the age groups. Conclusion: A majority of the sperm-head vacuoles showed a statistically significant difference among normal morphology groups. Therefore, we should consider the probability of the percentage of sperm-head vacuoles not increasing with age but with abnormal sperm morphology. A further study is required to clarify the effect of the sperm-head vacuoles on ART outcomes.
The aim of this study was to investigate the effect of sperm preservation according to the various kinds of commercially available semen extenders and to investigate the effect of sperm preservation according to the various temperature storages of fresh-extended porcine semen. To investigate the effect of sperm preservation according to the various kinds of commercially available semen extenders, porcine semens diluted in 3 semen extenders, Beltsville Thawing Solution(BTS), Androhep and Kiev, were cooled at $8^{\circ}C$ storage temperature with a controlled cooling rate of $2-4^{\circ}C$/h. Motility, progressive motility, normal sperm(%) and sperm morphology were assessed comparatively. In motility and progressive motility, Androhep extenders revealed better result than other extenders. In normal sperm(%) and sperm morphology, 3 semen extenders revealed similar results. To investigate the effect of sperm preservation according to the various temperature storages of fresh-extended porcine semen, porcine semens diluted in BTS extender, were cooled at 3 storage temperatures($8^{\circ}C$, $12^{\circ}C$ and $17^{\circ}C$) with a controlled cooling rate of $2-4^{\circ}C$/h. Motility, progressive motility, normal sperm(%) and sperm morphology were assessed comparatively. In motility and progressive motility, $8^{\circ}C$ treatment group revealed better result than $12^{\circ}C$ and $17^{\circ}C$ treatment groups. In normal sperm(%) and sperm morphology, 3 temperatures of treatment groups revealed similar results.
Nongbua, Thanapol;Utta, Apirak;Am-in, Nutthee;Suwimonteerabutr, Junpen;Johannisson, Anders;Morrell, Jane M
Asian-Australasian Journal of Animal Sciences
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제33권9호
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pp.1411-1420
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2020
Objective: The aim of study was to investigate the effects of season and single layer centrifugation (SLC) before cryopreservation on post-thaw bull sperm quality in Thailand. Methods: Semen was collected from 6 bulls (Bos indicus) in summer, rainy season and winter 2014 through 2016. Semen characteristics, sperm morphology, sperm kinematics, viability, chromatin structure and mitochondrial membrane were evaluated. Meteorological data were available from the local meteorological station; Results: Season had an effect on semen characteristics in the raw ejaculate, with higher proportions of normal spermatozoa and lower abnormalities in winter than in the other two seasons. Sperm kinematics, viability, DNA fragmentation index, and mitochondrial membrane potential were not different between seasons. Sperm samples selected by SLC had greater normal morphology and a lower proportion with bent tails than controls and higher values of progressive motility (PRO), beat cross frequency, linearity, straightness, wobble (WOB), and lower values of slow motility, velocity average path (VAP), velocity curved line, and amplitude of lateral head displacement than controls. In addition, SLC-selection had a favorable effect on PRO, VAP, and WOB that differed among seasons. Conclusion: Our results suggested that these bulls were well adapted to their location, with season having an effect on sperm morphology. Moreover, SLC could be used prior to cryopreservation, regardless of season, to enhance normal morphology and kinematics of bull sperm samples without adversely affecting other parameters of sperm quality. However, there was considerable variation among bulls in DNA fragmentation index, mitochondrial membrane potential and sperm viability. In addition, SLC had a positive effect on sperm morphology and sperm kinematics, which could be expected to influence fertility.
In male reproducible health, fertility and IVF (in-vitro fertilization), semen analysis has been most important. Semen analysis can be divided into concentration, motional and morphological analysis of sperm. The existing method which was developed earlier to analyze semen concentrated on the sperm motility analysis. To provide more useful and precise solutions for clinical problems such as infertility, semen analysis must include sperm morphological analysis. But the traditional tools for semen analysis are subjective, imprecise, inaccurate, difficult to standardize, and difficult to reproduce. Therefore, with the help of development of microcomputers and image processing techniques, we developed a new sperm morphology analyzer to overcome these problems. In this study the agreement on percent normal morphology was studied between different observers and a computerized sperm morphology analyzer on a slide-by-slide basis using strict criteria. Slides from 30 different patients from the SNUH andrology laboratory were selected randomly. Microscopic fields and sperm cells were chosen randomly and percent normal morphology was recorded. The ability of sperm morphology analyzer to repeat the same reading for normal and abnormal cells was studied. The results showed that there was no significant bias between two experienced observers. The limits of agreement were 4.1%${\sim}$-3.8%. The Pearson correlation coefficient between readers was 0.79. Between the manual and sperm morphology analyzer, the same findings were reported. In this experiments the slides were stained by two different methods, PAP and Diff-Quik staining methods. The limits of agreement were 7.2%${\sim}$-5.7% and 6.0%${\sim}$-6.3%, respectively. The Pearson correlation coefficients ware 0.76 and 0.91, respectively. The limits of agreement was tighter below 20% normal forms. In the experiments of repeatability, 52 cells stained by PAP and Diff-Quik staining methods were analyzed three times in succession. Estimating pairwise agreement, the kappa statistic for the pairs were 0.76, 0.81, 0.86, and 0.75, 0.88, 0.88 respectively. In this study it was shown that there was good agreement between manual and computerized assessment of normal and abnormal cells. The repeatability and agreement per slide of computerized sperm morphology analyzer was excellent. The computer's ability to classify normal morphology per slide is promising. Based on results obtained, this system can be of clinical value both in andrology laboratories and IVF units.
The recovery of epididymal sperm in animals is considered as one of the important tools to preserve high value or endangered species. However, there are no appropriate castrating indicators such as months of age in bull, sperm morphology, and motility, particularly in young Korean native bull (Hanwoo). Therefore, this study aimed to investigate sperm number, morphology, and motility of sperm in the epididymis tail of young Hanwoo bulls at 8 and 15 months of age. After castration, epididymal tails were collected and minced with blades to recover sperm. In experiments 1 and 2, sperm number, morphology, and motility were examined. Total number of sperm and percentage of normal sperm from bulls at 8 months of age was lower than that of bulls at 15 months of age after collection (P<0.05). Percentage of abnormal head, tail, proximal cytoplasmic droplet, dead and damaged acrosome of sperm from bulls at 8 months of age were higher than those of bulls at 15 months of age (P<0.05). In experiment 3, sperm motility from bulls at 8 and 15 months of age were examined before freezing and after thawing. Frozen-thawed sperm at 8 months of age showed low total motility and motile sperm with ${\geq}25{\mu}m/sec$ compared to those at 15 months of age and commercially-used sperm (P<0.05). In conclusion, sperm derived from the epididymal tail of bulls at 8 months of age showed high abnormal morphology and poor motility, which are not adequate for AI and IVF. On the other hand, sperm derived from the epididymal tail of bulls at 15 months of age showed high normal morphology and motility.
Spermatogenesis, ultrastructure, and sperm morphology of the Pacific oyster (Crassostrea gigas) were investigated with TEM and SEM. C. gigas sperm were primitive consisting of a head midpiece and tail. Sperm size (head and midpiece) was about 1.78 ${\mu}m$. Sperm morphology was similar to a sharp of a small water jar with a rough surface. Sperm had both anterior nuclear fossa (anf) and posterior nuclear fossa (pnf). Acrosome forms had a hat-like shape. The axial rod was projected in front of the acrosome. C. gigas sperm had four large mitochondria in the midpiece.
Kim, Hee-Sun;Kang, Moon Joo;Kim, Sung Ah;Oh, Sun Kyung;Kim, Hoon;Ku, Seung-Yup;Kim, Seok Hyun;Moon, Shin Yong;Choi, Young Min
Clinical and Experimental Reproductive Medicine
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제40권1호
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pp.23-28
/
2013
Objective: The aim of the present study was to examine the relationship among male age, strict morphology, and sperm chromatin structure and condensation. Methods: Sperm samples from a total of 100 men underwent semen analysis, and sperm chromatin structure and condensation were assessed with toluidine blue (TB) and aniline blue (AB) tests. Results: Prevalence of strict morphology of less than 4%, and abnormal sperm chromatin structure and condensation did not show any statistically significant differences according to male age (p=0.605, p=0.235, and p=0.080). No significant correlation was demonstrated among age of male partners, strict morphology, and abnormal sperm chromatin structure using TB and AB tests. However, abnormal sperm chromatin condensation was positively associated with sperm chromatin structure (r=0.594, p=0.000) and showed negative correlation with strict morphology (r=-0.219, p=0.029). Conclusion: The tests for sperm chromatin condensation showed a significant association with strict morphology. Further study is needed to elucidate the relationship between clinical outcome and sperm chromatin tests.
Objective: To investigate the effects of sperm morphology and their co-incubation with oocytes on the outcome of IVF and ICSI. Design: Strict morphology of washed sperm was assessed by Diff-Quick staining method before or after insemination. And the relationships between strict morphology and outcome (fertilization, embryo development and pregnancy) of IVF(with co-incubation) and ICSI (without co-incubation) were determined. Patients: Two-hundreds-and-sixty-three cycles of IVF and ninety-six cycles of ICSI were analyzed in order to clarify the influence of strict sperm morphology of spermatozoa on outcome of IVF and ICSI. These were divided into four groups. according to fertilization method and sperm morphology(Group 1: IVF, ${\geq}$12%, n:227; Group 2: IVF, <12%, n:36; Group 3: ICSI, ${\geq}$ 12%, n=48; Group 4: ICSI, <12%, n=48). Results: The fertilization rates of better morphology groups were higher than those of poor groups: Group 1(68.1%) > Group 2(62.1%), Group 3(78.1%) > Group 4(71.5%). There was no difference in embryo cleavage rates among four groups (>90%), Regarded with the good embryo rates, Group 1(56.8%) was significantly higher than Group 2(42.3%)(P<0.01), but there was no difference between Group 3(64.7%) and Group 4(61.2%). The pregnancy rates were also higher in better morphology groups as well as fertilization rates: Group 1(34.8%)> Group 2(16.7%)(p<0.05), Group 3(40.0%) > Group 4(23.0%)(p=0.08). Conclusion: Co-incubation with poor morphology sperm might adversely affect the quality of embryos. And strict sperm morphology may represent the ability to establish successful pregnancy. In short, the strict sperm morphology can be a good predictor of IVF and ICSI outcome.
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