231 SPERM SUBPOPULATIONS IN THE KOALA (PHASCOLARCTOS CINEREUS) EJACULATE

2009 ◽  
Vol 21 (1) ◽  
pp. 213
Author(s):  
N. Satake ◽  
S. D. Johnston ◽  
W. V. Holt

Koala semen contains a heterogeneous mixture of sperm morphotypes, mainly attributable to extreme degree of shape variability displayed by the hooked sperm head. By analogy with other species, we anticipate that the morphotypes may exhibit correspondingly different sperm-motility behaviors, largely caused by the differences in hydrodynamic interactions with the suspending media. This trend has been shown in human spermatozoa where motility behavior was demonstrably correlated with the sperm head morphology (Overstreet et al. 1981). In this study, we have investigated the heterogeneity of koala sperm motility profiles in semen in an effort to determine whether distinct sperm subpopulations within ejaculates are recognizable by the use of computer-assisted sperm motility analysis. Ejaculates from 5 males were collected by electroejaculation, then diluted and transported in Tris-citrate-glucose (TCG) diluent. Spermatozoa were washed through a 35–60% Percoll gradient to separate seminal plasma and the majority of the prostatic bodies from spermatozoa. Spermatozoa from the washed pellet were then diluted in TCG at 35°C, incubated for 10 min, and video recorded using a negative phase ×10 objective. Sperm motion parameters were then analyzed using the Hobson sperm tracker (Hobson Vision Systems, UK: Holt et al. 1996 J. Androl. 17, 587–596). Multivariate pattern analysis (PATN; CSIRO Australia; Abaigar 1999 Biol. Reprod. 60, 32–41) was used to distinguish 3 sperm subgroups, consistently shown in each ejaculate, within the data (1936 tracks × 6 kinetic parameters; VCL, VAP, MAD, BCF, ALH, LIN). After group allocation by PATN, all parameters showed significant differences between each of the groups (P < 0.0001). Group 1, approximately 25% of the sperm tracks, showed profiles of spermatozoa with fast, non-linear motility (VCL 106.88 ± 28.15; BCF 3.23 ± 3.81; LIN 14.08 ± 10.20). Group 2, approximately 27% of sperm tracks, showed profiles of fast, linear motility (VCL 63.92 ± 13.50; BCF 7.90 ± 3.42; LIN 28.10 ± 12.15). Group 3, 48% of sperm tracks, showed profiles of slow, non-linear or circular patterns of motility (VCL 39.05 ± 11.92; BCF 0.02 ± 0.35; LIN 5.15 ± 4.88). The recognition of 3 clearly identifiable subgroups supports our hypothesis that heterogeneity of sperm motility patterns exists within koala ejaculates. These may be a reflection of the heterogeneity in sperm-head morphotypes in koala semen, but that remains to be investigated in more detail. The clear distinctions between these groups, and the observation that all 3 subpopulations exist in each of the ejaculates, also suggest that the spermatozoa exhibit functional differences, possibly related to biochemical or maturational status. Many thanks to Dr. Michael Pyne and Dr. Vere Nicholson and their teams and animals at Currumbin Wildlife Sanctutary and Dreamwolrd QLD for all their help and support for the collection of samples.

2009 ◽  
Vol 50 (3) ◽  
pp. 306-311 ◽  
Author(s):  
Chih-Wei Wang ◽  
Chun-Jung Juan ◽  
Yi-Jui Liu ◽  
Hsian-He Hsu ◽  
Hua-Shan Liu ◽  
...  

Background: Although the ABC/2 formula has been widely used to estimate the volume of intracerebral hematoma (ICH), the formula tends to overestimate hematoma volume. The volume-related imprecision of the ABC/2 formula has not been documented quantitatively. Purpose: To investigate the volume-dependent overestimation of the ABC/2 formula by comparing it with computer-assisted volumetric analysis (CAVA). Material and Methods: Forty patients who had suffered spontaneous ICH and who had undergone non-enhanced brain computed tomography scans were enrolled in this study. The ICH volume was estimated based on the ABC/2 formula and also calculated by CAVA. Based on the ICH volume calculated by the CAVA method, the patients were divided into three groups: group 1 consisted of 17 patients with an ICH volume of less than 20 ml; group 2 comprised 13 patients with an ICH volume of 20 to 40 ml; and group 3 was composed of 10 patients with an ICH volume larger than 40 ml. Results: The mean estimated hematoma volume was 43.6 ml when using the ABC/2 formula, compared with 33.8 ml when using the CAVA method. The mean estimated difference was 1.3 ml, 4.4 ml, and 31.4 ml for groups 1, 2, and 3, respectively, corresponding to an estimation error of 9.9%, 16.7%, and 37.1% by the ABC/2 formula ( P<0.05). Conclusion: The ABC/2 formula significantly overestimates the volume of ICH. A positive association between the estimation error and the volume of ICH is demonstrated.


2020 ◽  
Vol 151 ◽  
pp. 01010
Author(s):  
Berlin P. Pardede ◽  
Iman Supriatna ◽  
Yudi Yudi ◽  
Muhammad Agil

This study aimed to analyze the effect of the age of bulls on sperm motility and DNA fragmentation and its impact on fertility. Ninety-six frozen semen straw from eight bulls were divided into four groups based on age (group-1: 5-6 years; group-2: 7-8 years; group-3: 9-10 years; group-4: 11-12 years). Total and progressive motility were detected by using computer-assisted semen analysis (CASA), while DNA fragmentation was detected by Toluidine blue staining. Over 500 artificial insemination services in the field were used for fertility rate analysis. The results of the analysis of total motility, progressive, and DNA fragmentation in all age groups still meet the minimum standard for artificial insemination programs. Analysis of progressive motility and DNA fragmentation showed significant differences in each age group (P<0.01), whereas analysis of total motility showed no significant differences in group-2 (7-8 years) and group-3 (9-10 years) (P>0.01). Increased age in bulls correlated significantly with increased sperm DNA fragmentation (P<0.01), decreased total and progressive motility (P<0,01), and potentially reduced the fertility rate (P<0.01). In conclusion, although the quality of frozen semen still meets the standards for artificial insemination programs, the age factor in bulls needs to be considered for achieving maximum fertility.


2015 ◽  
Vol 27 (7) ◽  
pp. 1049 ◽  
Author(s):  
Severine Larroze ◽  
Daniel B. Pickford ◽  
William V. Holt

We have developed and validated a computer-assisted sperm-motility assessment (CASA) method for use with the emerging amphibian model Silurana tropicalis. The testicular sperm-activation method was validated by analysing activation replicate coefficients of variation, effects of tracking time settings on velocity distributions and the relative partitioning of differentially motile sperm subpopulations between matched right and left testes. Two major sperm subpopulations were identified using multivariate pattern analysis and their relative frequencies were consistent between samples from matched right and left testes and from randomly drawn subsets of six frogs sampled from the total set of 16 frogs. The power of this approach for detecting treatment effects targeting the hypothalamic–pituitary–­gonadal axis was investigated by injecting a group of frogs with 100 IU human chorionic gonadotrophin (hCG) 2 h before sampling and comparing their sperm-subpopulation frequencies with non-injected controls. While parametric analysis across sperm samples failed to detect treatment effects, subpopulation analysis showed that hCG significantly increased the proportion of progressive and non-sinuous spermatozoa compared with controls (Chi square = 6.40, DF = 1, P = 0.011). This demonstrated the potential value of analysing objectively measured sperm behaviour as an endpoint.


2021 ◽  
Vol 16 (2) ◽  
pp. 171-181
Author(s):  
Sudipta Chowdhury ◽  
Samarendra Nath Banerjee

The genotoxicity of the ethanolic extract of betel nut was evaluated using sarcoma 180 tumour bearing mouse considering sperm motility, sperm viability, biochemical estimation of fructose in seminal fluid and sperm head morphology assays. Sperm head morphology was studied by H-E staining and Toluidine blue staining method. But Toluidine blue staining method is a reliable method to evaluate the DNA damage of sperms. Ethanolic BNE (betel nut extract) can suppress the percentage of sperm motility, sperm viability and seminal fructose level. In addition, it can also enhance the percentage of DNA damaged sperms. Moreover, histological sections of testes have been studied in control and BNE treated sarcoma 180 tumour bearing mice to highlight the potential toxic effect of BNE. The significant decreasing rate of seminal fructose concentration, sperm motility as well as viability and increasing rate of sperm head abnormality in different doses of treated series may be as a result of different toxic alkaloid ingredients present in BNE. Therefore, the results showed the potential of the BNE to induce different types of germ cell abnormalities in tumour bearing male mice.


Author(s):  
Yu Tsung Wu ◽  
Panos Papaspyridakos ◽  
Kiho Kang ◽  
Matthew Finkelman ◽  
Yukio Kudara ◽  
...  

The aims of this study were to evaluate the effect of (i) the different surgical guide designs and (ii) implant placement location on the accuracy of fully guided implant placement in single edentulous sites using an in vitro study model. Forty-five partially edentulous models were scanned and divided into three groups: group 1, tooth-supported full-arch surgical guide; group 2, three different tooth-supported shortened surgical guides (SSGs); and group 3, tooth-supported full-arch surgical guide with a crossbar. All surgical guides were printed and used for fully guided implant placement. A total of 180 implants (60 per group) were placed, and scanbodies were positioned on all models, and postoperative surface scan files (STL) files were obtained. Superimposition of preoperative and postoperative STL files was performed, and the accuracy of implant position was evaluated. The interaction between group and implant location was statistically significant for angle, 3D offset at the base, and at the tip (p&lt;0.001). The post-hoc tests showed a statistically significantly higher deviation for group 2 compared to group 3 for all outcomes for implants #4 (p&lt;0.05) and #7 (p&lt;0.05). There was also a statistically significant difference in all outcomes between groups 1 and 3 for implant #7 (p&lt;0.05). All surgical guide designs presented satisfactory performance with clinically acceptable levels of deviation. However, SSGs presented higher accuracy for guided implant placement in a single-edentulous site, whereas a full-arch surgical guide with a crossbar presented superior outcomes when two or more guided implants were placed simultaneously.


2010 ◽  
Vol 22 (1) ◽  
pp. 291
Author(s):  
M. Filliers ◽  
T. Rijsselaere ◽  
P. Bossaert ◽  
P. Anastasi ◽  
M. Hoogewijs ◽  
...  

Semen can be collected from tomcats by urethral catheterization (CT) after medetomidine administration, offering a novel approach to obtain sperm for in vitro fertilization. This study was designed to determine motion characteristics of CT sperm samples by means of computer-assisted sperm analysis (CASA) and to compare sperm quality parameters and in vitro fertilizing capacity of CT spermatozoa with those of spermatozoa retrieved after epididymal slicing (EP). Semen was collected in 17 adult cats by urethral catheterization as reported by Zambelli et al. (2008 Theriogenology 69, 485-490), after which the cat was orchidectomized. Motility (subjective and objective by means of CASA), morphology [eosin/nigrosin (E/N)], plasma membrane integrity (E/N and SYBR14-PI staining), and acrosomal status (FITC-PSA staining) of fresh CT and EP samples were evaluated and compared between both methods with a paired t-test or the Wilcoxon Rank test, dependent on normality of the data. In vitro maturation (24 h), fertilization (18 h), and culture (6h) of grade I to III oocytes were carried out as described by Pope et al. (2009 Theriogenology 71, 864-871). Twenty-four hours after in vitro insemination, fertilization rates were assessed for group 1 (CT; n = 148) and group 2 (EP; n = 159) presumptive zygotes. The distribution of presumptive zygotes between CT and EP over the different developmental stages was compared using Pearson chi-square test. Results showed that total and progressive motility as well as the percentage of normal spermatozoa were higher for EP sperm compared with CT sperm (P < 0.01). Epididymal sperm had a lower percentage of spermatozoa with an intact acrosome (P < 0.01), whereas CT sperm contained more spermatozoa with tail abnormalities (P < 0.01). Other sperm parameters did not differ (P > 0.05) between collection techniques. In group 1, 84% of in vitro-matured oocytes (metaphase II) were penetrated (40.2% cleaved, 24.4% with 2 pronuclei, 12.2% with >2 pronuclei, 7.3% with expanded sperm head), whereas in group 2, penetration rate was 88.5% (42.7% cleaved, 21.8% with 2 pronuclei, 16.7% with >2 pronuclei, 7.3% with expanded sperm head). No difference (P > 0.05) in in vitro fertilizing capacity between spermatozoa collected by means of the 2 methods was found. In conclusion, semen collection by means of CT yields fertilization results similar to epididymal slicing, despite the fact that several sperm variables were different. Because CT is repeatable and easy to perform and does not require a trained male/queen in heat, it may be preferable for routine IVF experiments with fresh spermatozoa. The first author is a research fellow of the Fund for Scientific Research-Flanders (Belgium).


2020 ◽  
Vol 28 (3) ◽  
pp. 230949902096567
Author(s):  
Jun Ho Nam ◽  
Suk-Kyoon Song ◽  
Myung-Rae Cho ◽  
Dae-Won Kang ◽  
Won-Kee Choi

Purpose: We have analyzed the surgical outcomes of primary total knee arthroplasty (TKA) using computer-assisted (CA) navigation in terms of postoperative coronal alignment depending on preoperative lateral femoral bowing. Methods: We conducted a retrospective study of patients who have undergone navigated primary TKA from January 2016 through January 2020. Two hundred and ninety-nine cases with lateral femoral bowing of 3° or less were assigned to group 1, 95 cases of lateral femoral bowing between 3° and 5° were assigned to group 2, and 89 cases with lateral femoral bowing of more than 5° were assigned to group 3. The postoperative mechanical hip–knee–ankle (mHKA) angle was measured from scanograms, which were taken 3 months after surgery. The appropriate range of coronal alignment was set as 0 ± 3°. Results: The number of outliers of mHKA occurred was 31 cases (10.4%) in group 1, 17 cases (17.9%) in group 2, and 17 cases (19.1%) in group 3. There was a significant correlation between the degree of lateral femoral bowing and the occurrence rate of mHKA outliers. Multiple variables logistic regression analysis showed occurrence rate of outliers in group 3 to be 2.04 times higher than group 1. After adjusting the patient’s age, sex, body mass index, and preoperative HKA deformity, the occurrence rate of outliers in group 3 was still 1.96 times higher than group 1. Conclusion: The benefit of CA navigation during TKA in obtaining coronal alignment within 0 ± 3° may be lessened when the preoperative lateral femoral bowing is severely advanced.


Reproduction ◽  
2001 ◽  
pp. 265-273 ◽  
Author(s):  
T Abaigar ◽  
M Cano ◽  
AR Pickard ◽  
WV Holt

Subjective and objective semen assessments were performed on 18 male Mohor gazelles (Gazella dama mhorr). Sperm motility assessments combined with sperm plasma membrane and acrosomal integrity evaluations were undertaken as part of a captive breeding programme. The primary objective was to test methodology for short-term preservation of gazelle semen for artificial insemination (storage in N-[Tris(hydroxymethyl)methyl]-2-aminoethanesulphonic acid-Tris diluent (TEST) for up to 96 h at 17 degrees C). However, the secondary objective was to investigate phenotypic and genotypic influences on semen quality within this small population, which was established in 1971 with only 12 genetic founders. Sperm motility was measured by computer-assisted semen assessment and the data were analysed using a pattern analysis technique to detect and quantify naturally occurring sperm subpopulations within the semen samples. Four sperm subpopulations distinguishable by their motion characteristics were detected. The relative frequencies of two subpopulations (population 2: highly motile, non-linear; and population 4: poorly motile, non-linear) in fresh semen were correlated with the maximum voltage used during electroejaculation. The frequency of subpopulation 2 was negatively correlated with maximum voltage (r = -0.875, P < 0.0001) and the frequency of subpopulation 4 was positively correlated (r = 0.727, P < 0.005). The frequencies of all subpopulations varied significantly among the animals sampled (chi-squared = 2577.6, degrees of freedom = 54, P < 0.0001) and subpopulation 4 was also correlated with body weight (r = -0.59, P < 0.005). Semen stored at 17 degrees C retained motility, plasma membrane and acrosomal integrity for 48 h, but these measures decreased thereafter. The frequency of a sperm subpopulation showing uncoordinated but active motility increased significantly over the first 48 h and then decreased.


Author(s):  
P. Bagavandoss ◽  
JoAnne S. Richards ◽  
A. Rees Midgley

During follicular development in the mammalian ovary, several functional changes occur in the granulosa cells in response to steroid hormones and gonadotropins (1,2). In particular, marked changes in the content of membrane-associated receptors for the gonadotropins have been observed (1).We report here scanning electron microscope observations of morphological changes that occur on the granulosa cell surface in response to the administration of estradiol, human follicle stimulating hormone (hFSH), and human chorionic gonadotropin (hCG).Immature female rats that were hypophysectcmized on day 24 of age were treated in the following manner. Group 1: control groups were injected once a day with 0.1 ml phosphate buffered saline (PBS) for 3 days; group 2: estradiol (1.5 mg/0.2 ml propylene glycol) once a day for 3 days; group 3: estradiol for 3 days followed by 2 days of hFSH (1 μg/0.1 ml) twice daily, group 4: same as in group 3; group 5: same as in group 3 with a final injection of hCG (5 IU/0.1 ml) on the fifth day.


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