Pogonias courbina sperm characteristcs in its first reproductive season.

Southern black drum (Pogonias courbina) is a species distributed along the western Atlantic Ocean, and it is the largest Sciaenidae observed in the coast of Rio Grande do Sul state, Brazil. However, it is listed as a vulnerable species at The IUCN Red List of Threatened Species™, and their fishing is prohibited. The objective of this study was to determine the sperm characteristics of P. courbina. Sperm samples of five young males (two-year-old fish) were collected through abdominal pressure. The sperm kinetics parameters were sperm motility (MOT) 10.7 ± 5.6%, curvilinear velocity (VCL) 120.07 ± 16.16 mm s ± 1, average path velocity (VAP) 75.64 ± 23.78 mm s ± 1, straight-line velocity (VSL) 62.49 ± 15.83 mm s ± 1, straightness (STR) 83.9 ± 5.3%, wobble (WOB) 61.9 ± 12.7%, beat cross frequency (BCF) 42.981 ± 4.627 Hz and progression (PRG) 1,805.4 ± 564.5 µm. The proportion of normal spermatozoa was 35.6 ± 6.1%. About the abnormalities observed, 22.7% occurred in the tail (short tail = 0.6 ± 0.5%, distally curled tail = 2.4 ± 1.6%, strongly curled tail = 1.9 ± 1.3%, broken tail = 7.9 ± 5.1%, folded tail = 5.5 ± 0.8%, loose tail = 4.4 ± 1.9%); 14.2% occurred in the head (degenerate head = 4.2 ± 1.6%, microcephaly = 1.8 ± 2.5%, loose head = 8.2 ± 2.1%) and 27.5% of the spermatozoa showed cytoplasmatic gouts (proximal gout = 20.0 ± 8.4%, distal gout = 7.5 ± 2.8%). Besides that, a correlation analysis was performed between sperm morphology and kinetics parameters, and the spermatozoa were measured for the morphometric parameters. There was a positive correlation between BCF and normal spermatozoa (r =  0.9269). A negative correlation occurred between BCF and loose head (r =  -0.9047); WOB and strongly curled tail (r =  -0.8911); and PROG and strongly curled tail (r =  -0.9191). The morphometric measures found for the head were length of 2.50 ± 0.21 µm and width of 2.12 ± 0.22 µm, and for the tail it was length of 37.97 ± 2.01 µm. It was possible to verify that the animals have sperm characteristics that indicate reproductive aptitude, but an abnormal behavior on sperm activation and high presence of the cytoplasmic gout abnormality indicates that the animals are not fully mature in their first reproductive season. This work contributes to a better understanding of the P. courbina spermatic parameters, what can be allies to recovery this species population in nature and promote its production in fish farms.

Sperm head morphometry and chromatin alterations in bulls with different conception rates.

The objective of this investigation was to analyze timed-AI conception rates (CRs) of different sires in light of their conventional semen quality parameters, sperm head morphometry, and chromatin alterations. Semen was collected in the field from six Angus bulls and used for the timed-AI of 890 suckled multiparous Nellore cows at a single farm. Semen batches were evaluated on the following in vitro parameters: sperm motility, concentration, and morphology, sperm head morphometry, and chromatin alteration types. The overall CR was 49% and Bulls 1 (43%) and 2 (40%) presented reduced (P < 0.05) pregnancies per AI compared to Bull 6 (61%), even though no differences were observed between their conventional semen quality parameters. Bull 1, however, presented higher (P = 0.0001) shape factor, smaller (P = 0.0025) antero-posterior symmetry, and elevated (P = 0.0141) Fourier 1 parameter, whereas Bull 2 exhibited a higher (P = 0.0023) percentage of chromatin alteration along the central axis of the sperm head. In conclusion, bulls with varying CRs may present sperm head morphometric differences and/or chromatin alterations while not presenting differences in conventional in vitro semen quality parameters. Although further studies are needed to elucidate the concrete implications of chromatin alterations on field fertility, sperm morphometric differences and chromatin alterations may be at least partially causative of the lower pregnancies per timed-AI of certain sires.

Effect of incubation and analysis temperatures on sperm kinematics and morphometrics during human semen analysis / Efecto de las temperaturas de incubación y análisis sobre la cinemática y la morfometría de los espermatozoides durante el análisis de semen humano

Introduction: Human semen analysis must be performed after the liquefaction of the ejaculate. This takes place about 30min after ejaculation and samples must be maintained in the lab during this time. The temperatures for this incubation and the final analysis of motility are crucial but seldom taken into account. This study aims to examine the effect of these temperatures on various sperm parameters both manually (sperm count, motility, morphology, viability, chromatin condensation and maturation and DNA fragmentation) and CASA (kinematics and morphometrics, using an ISAS®v1 CASA-Mot and CASA-Morph systems, respectively) analyzed. Methods: Seminal samples from thirteen donors were incubated for 10min at 37°C followed by additional 20min at either room temperature (RT, 23°C) or 37°C and then examined following WHO 2010 criteria. Results: The data obtained show that there were no significant differences (P>0.05) in the subjective sperm quality parameters with incubation temperature. On the other hand, the head sperm morphometric parameters were significantly higher after room temperature incubation showing, in addition, lower ellipticity (P<0.05). Furthermore, kinematic parameters were evaluated both at RT and 37°C for the two incubation temperatures. In general, the four temperature combinations showed that kinematic parameters followed this order: RT-RT Conclusions: Our results showed that temperature control during both incubation and analysis is needed for accurate semen analysis, recommending the use of 37°C during the entire process. (AU)

Introducción: El análisis de semen humano debe realizarse después de la licuefacción del eyaculado. Esto ocurre aproximadamente a los 30minutos después de la eyaculación. Las temperaturas para esta incubación y el análisis final de la motilidad son cruciales, pero rara vez se tienen en cuenta. Este estudio tiene como objetivo examinar el efecto de estas temperaturas en varios parámetros de los espermatozoides tanto de forma manual (recuento de espermatozoides, motilidad, morfología, viabilidad, condensación y maduración de la cromatina y fragmentación del ADN) como CASA (cinemática y morfometría, utilizando un CASA-Mot ISAS®v1 y Sistemas CASA-Morph, respectivamente) analizados. Métodos: Las muestras seminales de 13 donantes se incubaron durante 10minutos a 37°C, seguidas de 20minutos adicionales a temperatura ambiente (TA, 23°C) o a 37°C y luego se examinaron siguiendo los criterios de la OMS 2010. Resultados: Los datos obtenidos muestran que no hubo diferencias significativas (p>0,05) en los parámetros subjetivos de calidad del esperma con la temperatura de incubación. Por otro lado, los parámetros morfométricos de la cabeza de los espermatozoides fueron significativamente más altos después de la incubación a temperatura ambiente, mostrando, además, una elipticidad más baja (p<0,05). Además, los parámetros cinemáticos se evaluaron tanto a temperatura ambiente como a 37°C para las dos temperaturas de incubación. En general, las cuatro combinaciones de temperatura mostraron que los parámetros cinemáticos siguieron este orden: RT-RT < RT-37 < 37-37 < 37-RT (temperaturas de incubación y análisis, respectivamente). Conclusiones: Nuestros resultados mostraron que el control de la temperatura durante la incubación y el análisis es necesario para un análisis de semen preciso, recomendando el uso de 37°C durante todo el proceso. (AU)

Identification of Honey Bee Sperm Structures Following the use of Various Staining Techniques.

Introduction: Bees are currently artificially inseminated on a large scale for breeding and research purposes. The sperm of bees has a complex and varied structure, and determination of specific morphological defects in it is very difficult. Its comprehensive analysis by inspecting morphology and morphometry is an important tool for improving honey bee lines. The staining technique should interfere with the cells as little as possible while clearly showing the boundaries of the head and other elements. In this study, a comparative analysis of the morphometry of sperm was performed with various techniques for staining drone semen. Material and Methods: Semen was collected from 150 sexually mature Buckfast bee drones by artificially everting the copulatory organ. The morphology and morphometry of the sperm were assessed on slides prepared by three staining methods according to the protocols described online, using the Sperm Class Analyzer system. The lengths of the acrosome, nucleus, head in total, midpiece, tail without midpiece, tail with midpiece, and entire sperm were measured. Results: The most details of the drone sperm structure could be seen when stained with the eosin-nigrosin complex. This method made it possible to identify all structures and revealed the uneven distribution of sperm proteins in different parts of the tail. With the Sperm Stain method fewer details of the sperm structure were recognisable, and the fewest were with SpermBlue. Conclusion: The staining method, and thus the chemical reagents used, affect the dimensions of drone sperm. Given the great research potential of modified spermatozoa of insects, a standard for slide preparation for the evaluation of morphological and morphometric semen parameters should be established, as this would facilitate result comparison between laboratories and increase the value of morphological analysis of sperm for predicting and assessing fertility.

Effect of incubation and analysis temperatures on sperm kinematics and morphometrics during human semen analysis.

INTRODUCTION: Human semen analysis must be performed after the liquefaction of the ejaculate. This takes place about 30min after ejaculation and samples must be maintained in the lab during this time. The temperatures for this incubation and the final analysis of motility are crucial but seldom taken into account. This study aims to examine the effect of these temperatures on various sperm parameters both manually (sperm count, motility, morphology, viability, chromatin condensation and maturation and DNA fragmentation) and CASA (kinematics and morphometrics, using an ISAS®v1 CASA-Mot and CASA-Morph systems, respectively) analyzed. METHODS: Seminal samples from thirteen donors were incubated for 10min at 37°C followed by additional 20min at either room temperature (RT, 23°C) or 37°C and then examined following WHO 2010 criteria. RESULTS: The data obtained show that there were no significant differences (P>0.05) in the subjective sperm quality parameters with incubation temperature. On the other hand, the head sperm morphometric parameters were significantly higher after room temperature incubation showing, in addition, lower ellipticity (P<0.05). Furthermore, kinematic parameters were evaluated both at RT and 37°C for the two incubation temperatures. In general, the four temperature combinations showed that kinematic parameters followed this order: RT-RT

Bull spermatozoa selected by thermotaxis exhibit high DNA integrity, specific head morphometry, and improve ICSI outcome.

BACKGROUND: Sperm migration by thermotaxis is a guidance mechanism that operates along the oviduct and it has proved to be a valid method for selecting spermatozoa with low DNA fragmentation (SDF) in mice, humans, and stallions. This study aimed to analyse if bull spermatozoa could be selected by thermotaxis and to assess their quality in terms of SDF as well as determine the presence of a specific sperm subpopulation based on sperm morphometry and assess their fertilizing capacity by ICSI. METHODS: We used frozen-thawed sperm from 6 bulls and sperm selection by thermotaxis was performed with TALP medium supplemented with 25 mmol/L of HEPES and 5 mmol/L of caffeine. In these conditions, sperm selection was achieved, obtaining a net thermotaxis of 3.6%. Subsequently, we analysed the SDF of the migrated and not-migrated spermatozoa using the neutral COMET assay, and we evaluated the size of the sperm head using Hemacolor® staining with Motic Images Plus 3 software. Additionally, migrated and not-migrated spermatozoa by thermotaxis were used to fertilize bovine in vitro matured (IVM) oocytes by ICSI, a very inefficient procedure in cattle that is only successful when the oocyte is artificially activated. RESULTS: The results showed lower SDF (χ², P < 0.001, 13.3% reduction, n = 8) and lower head size parameters (length and width, P < 0.01; and perimeter and area, P < 0.001; n = 4) in those spermatozoa migrated in comparison to those not-migrated. The distribution of sperm subpopulations structure varied between groups, highlighting cluster 2, characterized by spermatozoa with small head size, and high ellipticity and elongated heads, as the most abundant in the thermotaxis migrated group. When performed ICSI (without oocyte artificial activation) with the thermotactic sperm, the blastocyst rate was 32.2% ± 9.3% in the group microinjected with the thermotactic spermatozoa vs. 8.3% ± 7.8% in the group of not-migrated sperm (χ², P < 0.05). CONCLUSION: Our results showed that bull sperm selection by thermotaxis has a much higher DNA integrity, small and elongated head size parameters, and different sperm subpopulation structure than the not-selected spermatozoa. Additionally, we evidenced that thermotactic spermatozoa improve ICSI success rates.

Influence of Prolactin Secretion Changes on Sperm Head Size and Freezability in Ibex and Mouflon.

Aim: This work examined the influence of induced changes in prolactin (PRL) secretion on sperm cryoresistance of ibex and the mouflon. Materials and Methods: PRL secretion was modified in a first experiment by the use of bromocriptine (BCR, dopamine agonist) during the non-breeding season, and in a second experiment by the use of sulpiride (SLP, dopamine D2-receptor antagonist) during the rutting season. Slow and ultra-rapid freezing protocols were used to cryopreserve sperm samples. Results: BCR decreased blood plasma PRL concentrations, whereas SLP increased them. Cryoresistance ratios (CRs) for curvilinear velocity (VCL), straight-line velocity (VSL), and average path velocity (VAP) in BCR-treated mouflons were lower than in controls using slow-freezing (p < 0.05), while CRs of motility and morphologically normal sperm of BCR-treated mouflons were greater than controls with ultra-rapid freezing (p < 0.05). BCR increased the head sperm dimensions in ibexes (p < 0.001); conversely, BCR decreased the head dimensions in mouflons (p < 0.001). CR-motility, CR-amplitude of lateral head displacement (ALH), CR-viability, and CR-acrosome integrity in SLP-treated mouflons were lower than in controls with slow-freezing (p < 0.01); CR-viability and CR-acrosome were lower than controls with ultra-rapid freezing (p < 0.05). In ibexes, CR-ALH was lower for SLP-treated (p < 0.05). SLP treatment increased head dimensions in ibexes (p < 0.001) but did not affect the sperm head of mouflons. Conclusion: Our findings show that high levels of blood plasma PRL negatively affect the cryoresistance of ibex and mouflon sperm.

Human kinematic and morphometric sperm subpopulation analysis using CASA technology: A new approach to spermatozoa classification / Análisis cinemático y morfométrico de subpoblaciones de espermatozoides humanos utilizando tecnología CASA: un nuevo enfoque para la clasificación de los espermatozoides

Introduction: Semen analysis is a clinical method aimed at determining the fertility of a male individual. The traditional subjective method lacks the reliability that can be achieved by computer-assisted sperm analysis (CASA) technology. Unfortunately, this technology has only been used when taking into consideration individually different sperm characteristics. The aim of this work is to present an integrative mathematical approach that considers different seminal variables to establish human sperm subpopulations. Methods: Samples were obtained from thirteen volunteers via masturbation and were analyzed by the routine subjective method and two objective systems, CASA Motility (CASA-Mot) and CASA Morphology (CASA-Morph). Results: Seminogram variables were reduced to three principal components (PC) showing two subpopulations. Kinematics and morphometric variables each rendered three PCs for four subpopulations. Conclusions: These results lay the foundations for future studies including different geographical, social, ethnic and age range conditions with the aim of achieving a definitive view of the human semen picture. (AU)

Introducción: El análisis de semen es el método clínico para determinar la fertilidad masculina. El método subjetivo tradicional carece de la fiabilidad, que se puede obtener con el uso de la tecnología del análisis de semen asistido por ordenador (CASA). Desafortunadamente, esta tecnología se ha venido utilizando únicamente teniendo en cuenta de forma independiente las diversas características de los espermatozoides. El objetivo del presente estudio es presentar una aproximación matemática que incluye diversas variables seminales para definir las posibles subpoblaciones espermáticas. Métodos: Las muestras se obtuvieron por masturbación de 13 voluntarios, que se analizaron de forma subjetiva, así como con 2 sistemas objetivos, para el análisis de la movilidad (CASA-Mot) y la morfología (CASA-Morph). Resultados: Tanto las variables cinemáticas como las morfométricas rindieron 3 componentes principales y 4 subpoblaciones. Conclusión: Estos resultados sientan las bases para estudios futuros que incluyan diferencias geográficas, sociales, étnicas o de rango de edad con el ánimo de obtener una definición concluyente sobre las características seminales de la especie humana. (AU)

Human kinematic and morphometric sperm subpopulation analysis using CASA technology: A new approach to spermatozoa classification.

INTRODUCTION: Semen analysis is a clinical method aimed at determining the fertility of a male individual. The traditional subjective method lacks the reliability that can be achieved by computer-assisted sperm analysis (CASA) technology. Unfortunately, this technology has only been used when taking into consideration individually different sperm characteristics. The aim of this work is to present an integrative mathematical approach that considers different seminal variables to establish human sperm subpopulations. METHODS: Samples were obtained from thirteen volunteers via masturbation and were analyzed by the routine subjective method and two objective systems, CASA Motility (CASA-Mot) and CASA Morphology (CASA-Morph). RESULTS: Seminogram variables were reduced to three principal components (PC) showing two subpopulations. Kinematics and morphometric variables each rendered three PCs for four subpopulations. CONCLUSIONS: These results lay the foundations for future studies including different geographical, social, ethnic and age range conditions with the aim of achieving a definitive view of the human semen picture.

Morphometry of Boar Spermatozoa in Semen Stored at 17 °C-The Influence of the Staining Technique.

The aim of the study was to assess the morphometry of sperm during storage of liquid boar semen at 17 °C. An attempt was also made to evaluate the suitability of three staining methods for assessment of boar sperm morphometry. The study was carried out on 20 Landrace boars. Semen was collected from the boars every 5 days by the manual method. Four ejaculates from each boar were analysed (80 ejaculates in total). Analyses were performed five times: at 1 h, 24 h, 48 h, 96 h, and 168 h after semen collection. Blisters with insemination doses were opened immediately before the analyses. From each insemination dose, smears were prepared for morphometric evaluation of sperm, which were stained by three methods (eosin-nigrosin-EN, eosin-gentian-EG, and SpermBlue-SB). Morphometric measurements of 15 randomly selected sperm with normal morphology were performed on each slide. The morphometric measurements included the following parameters: sperm head length, width, area, and perimeter; tail length; and total sperm length. The results of the morphometric measurements were used to calculate the head shape index. The morphometric dimensions of the sperm were shown to change during storage of semen at 17 °C. The extent of these changes, however, depended on the staining method used, as the three methods result in different morphometric dimensions of sperm, in the case of both the head and the tail. In the slides stained by the eosin-nigrosin method, the dimensions of the head and tail were smaller at every time of storage than in the slides stained by the SpermBlue and eosin-gentian methods.

What is the importance of sperm subpopulations?

The study of sperm subpopulations spans three decades. The origin, meaning, and practical significance, however, are less clear. Current technology for assessing sperm morphology (CASA-Morph) and motility (CASA-Mot) has enabled the accurate evaluation of these features, and there are many options for data classification. Subpopulations could occur as a result of the stage of development of each spermatozoon in the subpopulation. Spermatogenesis might contribute to the production of these subpopulations. Insights from evolutionary biology and recent molecular research are indicative of the diversity among male gametes that could occur from unequal sharing of transcripts and other elements through cytoplasmic bridges between spermatids. Sperm cohorts exiting the gonads would contain different RNA and protein contents, affecting the spermatozoon physiology and associations with the surrounding environmental milieu. Subsequently, these differences could affect how spermatozoa interact with the environmental milieu (maturation, mixing with seminal plasma, and interacting with the environmental milieu, or female genital tract and female gamete). The emergence of sperm subpopulations as an outcome of evolution, related to the reproductive strategies of the species, genital tract structures, and copulatory and fertilization processes. This kind of approach in determining the importance of sperm subpopulations in fertilization capacity should have a practical impact for conducting reproductive technologies, inspiring and enabling new ways for the more efficient use of spermatozoa in the medical, animal breeding, and conservation fields. This manuscript is a contribution to the Special Issue in memory of Dr. Duane Garner.

A Preliminary Study on Semen Collection, Its Evaluation, and Testicular and Sperm Morphometries in The Wild Proboscis Monkey (Nasalis Larvatus).

INTRODUCTION: The proboscis monkey (Nasalis larvatus) is an endangered species with a declining population. This article describes the first successful attempt at sperm collection and evaluation, and the testicular and sperm morphometries of the wild proboscis monkey in Sabah, Malaysia. MATERIAL AND METHODS: Eight semen collection procedures using electro-ejaculation and digital manipulation were conducted in three wild adult male proboscis monkeys. A total of 21 ejaculates were collected. The testicular biometry was measured with the aid of ultrasonography. Sample evaluation included semen volume and pH and sperm concentration, viability, and abnormality. The sperm morphometry was undertaken using phase contrast microscopy. RESULTS: The mean (±SD) total testicular volume of these animals was 5.77 cm3 (±1.58). Semen collection by electro-ejaculation resulted in an 84% success rate, while digital manipulation did not result in any ejaculation. Each animal showed different semen characteristics, where the volume was 5-540 µL, pH 8-9, and sperm concentration 0.041-83.00 ×106/mL. The percentage of abnormal sperm was high at 76.8% (±89.60), largely due to midpiece abnormality. Normal sperm had a spherical head and long tail with a head : midpiece : tail length ratio of 1 : 2: 8. CONCLUSION: The social status of these animals may contribute to the generally low quality of the semen. The techniques and data from this study are useful for future conservation and application of assisted reproductive technology in this species.

Spermatozoon head size - the main differentiating feature between spermatozoa of blue and white Arctic fox (Vulpes lagopus).

Morphology and sperm morphometry, this is an important determinant of male reproductive capacity. Morphometric data may provide relevant information in studies focused on evolutionary biology, sperm quality assessment, including prediction of the potential fertility, semen cryopreservation, or the effect of reprotoxicants. The paper presents the morphometric analysis of spermatozoa from two colour morphs of Arctic fox (Vulpes lagopus), and attempts to determine the relationship between selected quality indicators and dimensions and shape of spermatozoa. The research material consisted of ejaculates collected once by manual stimulation from 20 one-year-old Arctic foxes (10 individuals of the blue morph and 10 of the white morph). Ejaculates were analysed for standard parameters (volume, sperm concentration, total number of spermatozoa in the ejaculate) and used for the preparation of microscopic specimens. It was found that, the dimensions of spermatozoa from Arctic foxes depend on the male colour morphs. Spermatozoa from white Arctic foxes were significantly longer (by 1.82 µm) and had larger heads (0.32 µm longer and 0.15 µm wider) compared to spermatozoa from blue Arctic foxes (P<0.05). The interactions between particular sperm dimensions indicated the occurrence of gametes differing in shape. The all correlation coefficients between the morphometric traits of spermatozoa were statistically significant. Our research proved that in the blue Arctic foxes, sperm dimensions (tail length and total sperm length) can be related to the percentage of spermatozoa with primary changes (respectively: r = -0.68 and r = -0.75; at P <0.05). However, in the case of white Arctic foxes, these characteristics depend on the ejaculate volume (respectively: r = 0.65 and r = 0.68; at P <0.05).

Sperm morphometry is affected by increased inbreeding in the Retinta cattle breed: A molecular approach.

The effect of inbreeding depression on sperm motility is well documented, but its influence on sperm morphometry has been scarcely examined to date. Here, we combined the use of computer-assisted sperm morphometry analysis (CASMA) with a SNP-based genomic approach to determine and characterize the effect of inbreeding on the sperm shape of a highly inbred cattle population. We determined seven morphometric parameters on frozen-thawed sperm samples of 57 Retinta bulls: length (L, µm), width (W, µm), area (A, µm2 ), perimeter (P, µm), ellipticity (ELI; L/W), elongation (L-W)/(L + W) and perimeter-to-area shape factor (p2a; P2 /4 × π × A). The comparison of highly inbred (HI) and lowly inbreed (LI) individuals based on runs of homozygosity (ROH) inbreeding values (F ROH ) showed no differences between groups. An additional two-step unsupervised sperm subpopulation analysis based on morphometric parameters showed significant differences in the abundance of different sperm subpopulations between groups (p < 0.05). This analysis revealed that HI bulls harbored a higher percentage of narrow-head sperm as opposed to the higher percentage of large- and round-headed sperm detected in LI. A further genomic characterization revealed 23 regions differentially affected by inbreeding in both groups, detecting six genes (SPAG6, ARMC3, PARK7, VAMP3, DYNLRB2, and PHF7) previously related to different spermatogenesis-associated processes.

Spermatogenic Activity and Sperm Traits in Post-Pubertal and Adult Tomcats (Felis catus): Implication of Intra-Male Variation in Sperm Size.

Tomcats are considered to be adults at 1 year of age, although many reach sexual maturity at an earlier age. Nevertheless, we still know little about whether the spermatogenic activity and sperm quality of mature under one-year-old tomcats differ from those of tomcats that are over one-year-old. This study aims to evaluate the spermatogenic activity, sperm traits, and their relationships in mature tomcats at two different ages. Sixteen tomcats showing complete spermatogenesis and spermatozoa in their epididymal caudae were used and classified according to their age as post-pubertal (<1 year old) or adult (˃1 year old). Our results show that adult cats have higher epididymal sperm concentration and lower coefficient of variation in sperm head width and ellipticity than post-pubertal cats. However, they do not differ in their testicular and epididymal mass, spermatogenesis, and sperm traits such as motility, mitochondrial activity, morphology, morphometry, as well as plasma membrane, acrosome, and DNA integrity. Reduced intra-male variation of sperm head ellipticity is associated with higher testis mass, epididymis mass, and sperm concentration. Interestingly, low intra-male variation in sperm head size is associated with increased Sertoli cell function and reduced post-meiotic germ cell loss. These findings increase our knowledge about feline reproductive physiology and provide new insights into the functional significance of low intra-male variation in sperm size and shape in tomcats.

New insights in Cercopithecinae spermatozoa.

Sperm morphometric and morphologic data have been shown to represent useful tools for monitoring fertility, improving assisted reproduction techniques and conservation of genetic material as well as detecting inbreeding of endangered primates. We provide here for the first time sperm morphologic and morphometric data from Cercopithecus neglectus, Cercopithecus cephus, Papio papio and critically endangered Cercopithecus roloway, as well as comparative data from other Cercopithecinae species, i.e. Allochrocebus lhoesti, Mandrillus sphinx and Papio anubis. Following collection from the epididymis, spermatozoa were measured for each species for the following parameters: head length, head width, head perimeter, head area, midpiece length and total flagellum length, and the head volume, ellipticity, elongation, roughness and regularity were then calculated. Our data are consistent with both the general morphology and the morphometric proportions of Cercopithecinae sperm. Some specificities were observed, with C. cephus displaying a narrow head (width = 2.76 ± 0.26 µM) and C. roloway displaying a short midpiece (6.65 ± 0.61 µM). This data set represents an important contribution, especially for Cercopithecus roloway, one of the most endangered monkeys in the world, and further data on additional specimens coupled to data on mating systems and reproductive ecology should allow a better understanding of the mechanisms underlying these morphological differences across primate species.

Evaluation of the Morphometry of Sperm from the Epididymides of Dogs Using Different Staining Methods.

Evaluation of sperm morphometry is an important criterion in the diagnosis of a male animal's suitability for breeding. The aim of the study was to evaluate the morphometry of sperm from the epididymides of dogs subjected to routine castration using various staining methods. The study was carried out on semen collected from ten healthy dogs. Gonads were obtained from each dog during routine castration at a veterinary surgery. Then, the epididymides (caput, corpus, cauda) were isolated from the gonads, semen was collected from them and microscope slides were prepared. The slides for evaluation of sperm morphometry were prepared by four methods: DiffQuik, SpermBlue, eosin-nigrosin and eosin-gentian. A total of 2400 sperm were analyzed (240 sperm from the dog). The sperm collected from the caput and corpus of the epididymis were found to have larger heads and tails than those collected from the cauda of the epididymis. The staining method was shown to affect the morphometry of sperm taken from the epididymides of dogs. The staining methods differentiate the dimensions of the head of sperm in different parts of the epididymis but do not affect the length of the sperm tail. The occurrence of differences in the head dimensions of sperm may be linked to the use of different fixatives and chemical reagents in the staining procedure. Sperm stained by the EN method had the smallest head and tail dimensions. The greatest head area was noted in the sperm stained by the EG method. In the slides stained by the SB method, the sperm heads were relatively long but narrow. The methods used are suitable for the evaluation of sperm structure, and the possibility of using all four methods enables a full characterization of sperm collected from the caput, corpus and cauda of the epididymides of dogs.

Sperm head morphometry and chromatin condensation are in constant change at seminiferous tubules, epididymis, and ductus deferens in bulls.

The aim of this study was to evaluate the sperm head morphometry and chromatin condensation at different regions of the reproductive tract in bulls. Sperm smears of seminiferous tubules (ST), epididymis head (EH), body (EB), and tail (ET), and ductus deferens (DD) were stained with toluidine blue. Afterwards, the sperm head morphometry and chromatin alteration types were evaluated by a computational image analysis. Overall, spermatozoa of ST had lower (P < 0.05) area (A), perimeter (P), width (W), length (L), ellipticity (E), and Fourier harmonics (F0, F1, and F2). The chromatin decondensation (CD) and heterogeneity (CH) were higher (P < 0.05) in the ST region and decreased (P < 0.0001) during the migration along the reproductive tract (ST - DD direction). Considering the factors extracted (Factors 1 and 2) by the principal component analysis, the parameters A, P, W, L, and F0 were responsible for ∼36% of the Factor 1, while the E, F0, F1, and anterior-posterior symmetry (APS) contributed ∼27% to Factor 2. Both, CD and CH were associated with Factor 1 in the EH and ET regions and Factor 2 in the ST. Also, a well-defined difference between sperm heads collected from the ST and DD regions was observed by canonical analysis. The distribution of each chromatin alteration type was recorded. The proportion of normal sperm was lower (P < 0.05) in ST compared to other regions. Moreover, the chromatin influenced the morphometry and sperm heads with whole chromatin alteration type showed a smaller (P < 0.05) A, P, W, L, and E. In summary, the epididymal maturation is important for chromatin compaction and final morphometry of the sperm head. Also, the identification and quantification of the sperm chromatin condensation in different regions of reproductive tract can be used as potential biomarkers to predict the fertility in bulls.

Evaluation of sperm morphometry of rabbits (Oryctolagus cuniculus f. domesticus).

Sperm morphology and morphometry are considered parameters in fertility diagnosis. They are especially important in the case of species for which there is no standard with respect to morphometric sperm parameters. It is then crucial to apply the staining technique that has the least influence on the sperm structure and provides the most detailed image, so as to enable measurements. The aim of the research was to assess the morphometric parameters of rabbit sperm using silver nitrate staining. The staining process revealed a detailed image of the spermatozoon head and tail, thus enabling precise measurements. From these basic morphometric parameters, four additional shape indices characterizing the sperm head were calculated: ellipticity, elongation, roughness and regularity. These parameters more precisely characterize the shape of the sperm head. Silver nitrate staining can be used as an independent technique in assessment of sperm structure or to supplement routine diagnostics.

Effect of deep transfer and multi-task learning on sperm abnormality detection.

Analyzing the abnormality of morphological characteristics of male human sperm has been studied for a long time mainly because it has many implications on the male infertility problem, which accounts for approximately half of the infertility problems in the world. Yet, detecting such abnormalities by embryologists has several downsides. To clarify, analyzing sperms through visual inspection of an expert embryologist is a highly subjective and biased process. Furthermore, it takes much time for a specialist to make a diagnosis. Hence, in this paper, we proposed two deep learning algorithms that are able to automate this process. The first algorithm uses a network-based deep transfer learning approach, while the second technique, named Deep Multi-task Transfer Learning (DMTL), employs a novel combination of network-based deep transfer learning and multi-task learning to classify sperm's head, vacuole, and acrosome as either normal or abnormal. This DMTL technique is capable of classifying all the aforementioned parts of the sperm in a single prediction. Moreover, this is the first time that the concept of multi-task learning has been introduced to the field of Sperm Morphology Analysis (SMA). To benchmark our algorithms, we employed a freely-available SMA dataset named MHSMA. During our experiments, our algorithms reached the state-of-the-art results on the accuracy, precision, and f0.5, as well as other important metrics, such as the Matthews Correlation Coefficient on one, two, or all three labels. Notably, our algorithms increased the accuracy of the head, acrosome, and vacuole by 6.66%, 3.00%, and 1.33%, and reached the accuracy of 84.00%, 80.66%, and 94.00% on these labels, respectively. Consequently, our algorithms can be used in health institutions, such as fertility clinics, with further recommendations to practically improve the performance of our algorithms.