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Spermiogram and sperm head morphometry assessed by multivariate cluster analysis results during adolescence （12-18 years） and the effect of varicocele

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Asian Journal of Andrology 2016年第6期18卷 824-830页

作者： Fernando Vasquez Caries Soler Patricia Camps anthony valverde Almudena Garcia-Molina Research Group on Reproductive and Sexual Health Health Sciences Division Universidad del Norte EO. Box 1569 Barranquilla Colombia Department of Functional Biology and Physical Anthropology University of Valencia 46100 Burjassot Valencia Spain R+D Department Proiser R+D Scientific Park University of Valencia 46980 Paterna Valencia Spain Technological Institute of Costa Rica Agronomy School Animal Reproduction Department San Carlos Campus 223-21001 San Carlos Costa Rica

This work evaluates sperm head morphometric characteristics in adolescents from 12 to 18 years of age, and the effect of varicocele. Volunteers between 150 and 224 months of age （mean 191, n = 87）, who had reached oigarche by 12 years old, were recruited in the area of Barranquilla, Colombia. Morphometric analysis of sperm heads was performed with principal component （PC） and discriminant analysis, Combining seminal fluid and sperm parameters provided five PCs： two related to sperm morphometry, one to sperm motility, and two to seminal fluid components. Discriminant analysis on the morphometric results of varicocele and nonvaricocele groups did not provide a useful classification matrix. Of the semen-related PCs, the most explanatory （40%） was related to sperm motility. Two PCs, including sperm head elongation and size, were sufficient to evaluate sperm morphometric characteristics. Most of the morphometric variables were correlated with age, with an increase in size and decrease in the elongation of the sperm head. For head size, the entire sperm population could be divided into two morphometric subpopulations, SP1 and SP2, which did not change during adolescence. In general, for varicocele individuals, SP1 had larger and more elongated sperm heads than SP2, which had smaller and more elongated heads than in nonvaricocele men. In summary, sperm head morphometry assessed by CASA-Morph and multivariate cluster analysis provides a better comprehension of the ejaculate structure and possibly sperm function. Morphometric analysis provides much more information than data obtained from conventional semen analysis.

关键词： adolescence CASA-Morph system seminal quality sperm head morphometry spermiogram subpopulation

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Afterword to Sperm morphometrics today and tomorrow special issue in Asian Journal of Andrology

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Asian Journal of Andrology 2016年第6期18卷 895-897页

作者： Caries Soler Trevor G Cooper anthony valverde Jesfis L Yanliz Department of Functional Biology and Physical Anthropology University of Valencia 46100 Burjassot Spain Department of R+D Proiser R+D Science Park University of Valencia 46980 Paterna Spain Tuen Mun New Territories Hong Kong SAR China Department of Animal Production and Food Science Higher Polytechnic School of Huesca University of Zaragoza Ctra Cuarte S/N 22071 Huesca Spain

The problems associated with the subjective assessment of human sperm morphology have been well aired in another Asian Journal of Andrology Special Issue1 that marked the publication of the 5th edition of the WHO Semen analysis manual, and contrary views have subsequently been presented） However, the vagaries of the eye-brain system in assessing whether a sperm head is large or small can be eliminated by objective assessment where definitive structures are defined by their dimensions. These can then be classified automatically into as many categories as the data permit, conventionally on the basis of preset upper and lower limits, but also by more comprehensive analysis as discussed here.

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Morphometry and subpopulation structure of Holstein bull spermatozoa： variations in ejaculates and cryopreservation straws

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Asian Journal of Andrology 2016年第6期18卷 851-857页

作者： anthony valverde Hector Arenan Maria Sancho Jesus Contell Jesus Yaniz Alejandro Fernandez Caries Soler Technological Institute of Costa Rica San Carlos Campus S Xenetica Fontao Fontao-Esperante 27210 Lugo Spain R+D Department Proiser R+D Scientific Park University of Valencia 46980 Paterna Spain Department of Animal Production and Food Science Politechnical School of Huesca University of Zaragoza 22071 Huesca Spain Department of Functional Biology and Physical Anthropology University of Valencia 46100 Burjassot Spain

Sperm quality is evaluated for the calculation of sperm dosage in artificial reproductive programs. The most common parameter used is motility, but morphology has a higher potential as a predictor of genetic quality. Morphometry calculations from CASA-Morph technology improve morphological evaluation and allow mathematical approaches to the problem. Semen from 28 Holstein bulls was collected by artificial vagina, and several ejaculates were studied. After general evaluation, samples were diluted, packaged in 0.25 ml straws, and stored in liquid nitrogen. Two straws per sample were thawed, and slides were processed and stained with Diff-Quik. Samples were analyzed by a CASA-Morph system for eight morphometric parameters. In addition to the ＂classical＂ statistical approach, based on variance analysis （revealing differences between animals, ejaculates, and straws）, principal component （PC） analysis showed that the variables were grouped into PC1, related to size, and PC2 to shape. Subpopulation structure analysis showed four groups, namely, big, small, short, and narrow from their dominant characteristics, representing 31.0%, 27.3%, 24.1%, and 17.7% of the total population, respectively. The distributions varied between animals and ejaculates, but between straws, there were no differences in only four animals. This modern approach of considering an ejaculate sperm population as divided into subpopulations reflecting quantifiable parameters generated by CASA-Morph systems technology opens a new view on sperm function. This is the first study applying this approach to evaluate different ejaculates and straws from the same individual. More work must be done to improve seminal dose calculations in assisted reproductive programs.

关键词： cryopreservation Holstein bull sperm morphometry subpopulations

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Morphometric comparison by the ISAS CASA-DNAf system of two techniques for the evaluation of DNA fragmentation in human spermatozoa

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Asian Journal of Andrology 2016年第6期18卷 835-839页

作者： Sara Sadeghi Almudena Garcia-Molina Ferran Celma anthony valverde Sogol Fereidounfar Caries Soler Department of Functional Biology and Physical Anthropology University of Valencia 46100 Burjassot Valencia Spain R+D Department Proiser R+D Scientific Park Universitat de Valencia 46980 Paterna Valencia Spain Technological Institute of Costa Rica San Carlos Campus School of Agronomy 223-21001 Alajuela Costa Rica Department of Botany Kharazmi University Tehran Iran

DNA fragmentation has been shown to be one of the causes of male infertility, particularly related to repeated abortions, and different methods have been developed to analyze it. In the present study, two commercial kits based on the SCD technique （Halosperm~ and SDFA） were evaluated by the use of the DNA fragmentation module of the ISAS vl CASA system. Seven semen samples from volunteers were analyzed. To compare the results between techniques, the Kruskal-Wallis test was used. Data were used for calculation of Principal Components （two PCs were obtained）, and subsequent subpopulations were identified using the Halo, Halo/Core Ratio, and PC data. Results from both kits were significantly different （P 〈 0.001）. In each case, four subpopulations were obtained, independently of the classification method used. The distribution of subpopulations differed depending on the kit used. From the PC data, a discriminant analysis matrix was obtained and a good a posterioriclassification was obtained （97.1% for Halosperm and 96.6% for SDFA）. The present results are the first approach on morphometric evaluation of DNA fragmentation from the SCD technique. This approach could be used for the future definition of a classification matrix surpassing the current subjective evaluation of this important sperm factor.

关键词： DNA fragmentation Halosperm morphometry principal component analysis SDFA subpopulation

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Sperm kinematic, head morphometric and kinetic- morphometric subpopulations in the blue fox （Alopex lagopus）

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Asian Journal of Andrology 2017年第2期19卷 154-159页

作者： Caries Soler Jesus Contell Lorena Bori Maria Sancho Almudena Garcia-Molina anthony valverde Jan Segarvall Department of Functional Biology and Physical Anthropology University of Valencia 46100 Burjassot Spain R+D Department Proiser R+D Scientific Park University of Valencia 46980 Paterna Spain Technological Institute of Costa Rica San Carlos Campus School of Agronomy 223-21001 Alajuela Costa Rica Finnish Fur Breeders Association ProFur 65101 Vaasa Finland

This work provides information on the blue fox ejaculated sperm quality needed for seminal dose calculations. Twenty semen samples, obtained by masturbation, were analyzed for kinematic and morphometric parameters by using CASA-Mot and CASA-Morph system and principal component （PC） analysis. For motility, eight kinematic parameters were evaluated, which were reduced to PC1, related to linear variables, and PC2, related to oscillatory movement. The whole population was divided into three independent subpopulations： SP1, fast cells with linear movement; SP2, slow cells and nonoscillatory motility; and SP3, medium speed cells and oscillatory movement. In almost all cases, the subpopulation distribution by animal was significantly different. Head morphology analysis generated four size and four shape parameters, which were reduced to PC 1, related to size, and PC2, related to shape of the cells. Three morphometric subpopulations existed. SPI： large oval cells; SP2： medium size elongated cells; and SP3. small and short cells. The subpopulation distribution differed between animals. Combining the kinematic and morphometric datasets produced PC1, related to morphometric parameters, and PC2, related to kinematics, which generated four sperm subpopulations - SPI.. high oscillatory motility, large and short heads; SP2; medium velocity with small and short heads; SP3. slow motion small and elongated cells; and SP4. high linear speed and large elongated cells. Subpopulation distribution was different in all animals. The establishment of sperm subpopulations from kinematic, morphometric, and combined variables not only improves the well-defined fox semen characteristics and offers a good conceptual basis for fertility and sperm preservation techniques in this species, but also opens the door to use this approach in other species, included humans.

关键词： integration of motility and morphology principal component analysis sperm morphometry subpopulation

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Updating semen analysis:a subpopulation approach

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Asian Journal of Andrology 2020年第1期22卷 118-119页

作者： Almudena Garcia-Molina anthony valverde Daznia Bompart Carina Caldeira Alberto Vendrell Carles Soler Proiser R+D S.L. Scientific ParkUniversitat de ValenciaPaterna 46980ValenciaSpain Department of Functional Biology and Physical Anthropology University of ValenciaBurjassot 46100ValenciaSpain Technological Institute of Costa Rica San Carlos CampusSchool of AgronomyAlajuela 223-21001Costa Rica

Dear Editor,Reproductive problems affect 8%-12%of human populations worldwide;40%-50%of all infertility cases are due to the male,and up to 2%of men have suboptimal sperm quality.

关键词： dating cases infertility

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Dog sperm head morphometry： its diversity and .=volution

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Asian Journal of Andrology 2017年第2期19卷 149-153页

作者： Caries Soler Ana Alambiaga Maria A Marti Almudena Garcia-Molina anthony valverde Jesus Contell Marcos Campos Department of Functional Biology and Physical Anthropology University of Valencia 46100 Burjassot Spain R+D Department Proiser R+D Scientific Park University of Valencia 46980 Paterna Spain 3Technological Institute of Costa Rica San Carlos Campus School of Agronomy 223-21001 Alajuela Costa Rica Department of Medicine and Animal Surgery University CardenaI-Herrera-CEU 46113 Moncada Valencia Spain Global Veterinaria (Reprovalcan) 46500 Sagunto Valencia Spain Clinica Veterinaria Sangueso 46009 Valencia Spain

Dogs have been under strong artificial selection as a consequence of their relationship with man. Differences between breeds are evident that could be reflected in seminal characteristics. The present study was to evaluate differences in sperm head morphometry between seven well-defined breeds of dog： the British Bulldog, Chihuahua, German Shepherd, Labrador Retriever, Spanish Mastiff, Staffordshire Terrier, and Valencian Rat Hunting dog. Semen samples were obtained by masturbation and smears stained with Diff-Quik. Morphometric analysis （CASA-Morph） produced four size and four shape parameters. Length, Ellipticity, and Elongation showed higher differences between breeds. MANOVA revealed differences among all breeds. Considering the whole dataset, principal component analysis （PCA） showed that PC1 was related to head shape and PC2 to size. Procluster analysis showed the British Bulldog to be the most isolated breed, followed by the German Shepherd. The PCA breed by breed showed the Chihuahua, Labrador Retriever, Spanish Mastiff, and Staffordshire Terrier to have PC1 related to shape and PC2 to size, whereas the British Bulldog, Valencia Rat Hunting dog, and German Shepherd had PC1 related to size and PC2 to shape. The dendrogram for cluster groupings and the distance between them showed the British Bulldog to be separated from the rest of the breeds. Future work on dog semen must take into account the large differences in the breeds＇ sperm characteristics. The results provide a base for future work on phylogenetic and evolutionary studies of dogs, based on their seminal characteristics.

关键词： Canis familiaris cluster analysis diversity sperm morphometry

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