Effect of different sizes of nanocopper particles on rainbow trout (Oncorhynchus mykiss W.) spermatozoa motility kinematics.

In recent years, nanocopper (Cu NPs) has gained attention due to its antimicrobial properties and potential for industrial, agricultural, and consumer applications. But it also has several effects on the aquatic environment. Widespread use of various nanoproducts has raised concerns about impacts of different nanoparticle size on environment and biological objects. Spermatozoa is a model for studying the ecotoxic effects of pollutants on cells and organisms. This study aimed to investigate the effects of different sizes of copper nanoparticles on rainbow trout spermatozoa motility, and to compare their effects with copper ionic solution. Computer assisted sperm analysis (CASA) was used to detect movement parameters at activation of gametes (direct effect) with milieu containing nanocopper of primary particle size of 40-60, 60-80 and 100 nm. The effect of the elements ions was also tested using copper sulfate solution. All products was prepared in concentration of 0, 1, 5, 50, 125, 250, 350, 500, 750, and 1000 mg Cu L-1. Six motility parameters were selected for analysis. The harmful effect of Cu NPS nanoparticle was lower than ionic form of copper but the effect depends on the motility parameters. Ionic form caused complete immobilization (MOT = 0 %, IC100) at 350 mg Cu L-1 whilst Cu NPs solution only decreased the percentage of motile sperm (MOT) up to 76.4 % at highest concentration tested of 1000 mg Cu L-1 of 40-60 nm NPs. Cu NPs of smaller particles size had more deleterious effect than the bigger one particularly in percentage of MOT and for curvilinear velocity (VCL). Moreover, nanoparticles decrease motility duration (MD). This may influence fertility because the first two parameters positively correlate with fertilization rate. However, the ionic form of copper has deleterious effect on the percentage of MOT and linearity (LIN), but in some concentrations it slightly increases VCL and MD.

Siamese Neural Network for Keystroke Dynamics-Based Authentication on Partial Passwords.

The paper addresses issues concerning secure authentication in computer systems. We focus on multi-factor authentication methods using two or more independent mechanisms to identify a user. User-specific behavioral biometrics is widely used to increase login security. The usage of behavioral biometrics can support verification without bothering the user with a requirement of an additional interaction. Our research aimed to check whether using information about how partial passwords are typed is possible to strengthen user authentication security. The partial password is a query of a subset of characters from a full password. The use of partial passwords makes it difficult for attackers who can observe password entry to acquire sensitive information. In this paper, we use a Siamese neural network and n-shot classification using past recent logins to verify user identity based on keystroke dynamics obtained from the static text. The experimental results on real data demonstrate that keystroke dynamics authentication can be successfully used for partial password typing patterns. Our method can support the basic authentication process and increase users' confidence.

The Effect of Copper and Copper Oxide Nanoparticles on Rainbow Trout (Oncorhynchus mykiss W.) Spermatozoa Motility after Incubation with Contaminants.

The study aimed to analyse the effect of copper nanoparticles of similar particle size of Cu and CuO and copper ions (CuSO4) on the motility parameters of rainbow trout spermatozoa after long-term exposure and compare its harmful effect. Nanoproducts of Cu and CuO (Cu NPs, CuO NPs) of primary particle size around 50 nm and ionic solution of CuSO4 were used for the study. Suspension of concentrations 0, 1, 5, 10, 25, and 50 mg Cu·L-1 of Cu NPs, CuO NPs, and CuSO4 was dissolved in an artificial seminal plasma. Milt was mixed with the prepared solution and stored in a fridge, at 6 °C, for 96 h. At the defined incubation time, spermatozoa were activated for movement, and six motility parameters were evaluated using an automated system (CASA). Increasing concentrations of Cu NPs, CuO NPs, and CuSO4 in an incubation medium in parallel decreased the percentage of motile sperm (MOT). The effect of Cu NPs and ionic copper on MOT was more deleterious than that of CuO NPs. Copper products slightly increased the velocity (VCL) compared to the control, particularly up to 24 h of storage. Linearity (LIN) was improved by three tested products, particularly CuO NPs. Generally, the motility duration was prolonged when the sperm was incubated with copper products compared to the control. Nanoproducts made from different compounds of the same elements of similar particle size have a different effect on cells. Cu NPs were more harmful than CuO NPs. The effect of Cu NPs was similar to an ionic form of CuSO4. When incubated, the copper nanoproducts and ionic form exert a slightly positive effect on spermatozoa velocity, linearity, and motility duration, particularly up to 24 h of storage.

Survival of Embryos and Fry of Sea Trout (Salmo trutta m. trutta) Growing from Eggs Exposed to Different Concentrations of Selenium during Egg Swelling.

Adequate selenium (Se) availability enhances the health and growth of organisms, but overdose of it can be harmful and pathogenic. The study's objective was to analyse the impact of short-term exposure of sea trout fertilised eggs to inorganic selenium (SeO2) at concentrations from 0 to 32 mg Se L-1 to find the optimal and toxic dose of Se on early fish development. Se accumulated in the body, embryos' survival rate, and growth in the first four months of life was examined. Swelling of fertilised eggs in water supplemented with Se at a concentration from 0.5 to 8 mg Se L-1 was associated with a slightly positive impact on the hatching rate. At higher Se concentration, a harmful effect on the survival of the embryo was observed. The survival of fry was similar in all groups, while the fry length and weight correlated positively with Se concentration in its body. Immersion of fertilised eggs in water enriched with Se during egg swelling can constitute a method to supplement the element to non-feeding stages of fish. In selenium-poor areas, this innovative method can be implemented in aquaculture to improve breeding outcomes. Se concentration should be adjusted to the chemical compound, fish species, and Se's content in the yolk.

Reproductive Potential of Stone Moroko (Pseudorasbora parva, Temminck et Schlegel, 1846) (Teleostei: Cypriniformes: Gobionidae) Inhabiting Central Europe.

Similar to other invasive species, stone moroko is extending its global distribution. The present study aimed to assess the reproductive potential of stone moroko fish in a new habitat in Poland based on analysing the sexual cycle and fecundity. Fish morphometric data, age, and gonadal structures were analysed. Fish age ranged from 0+ to 5+ years. Most females and males (93% and 60%, respectively) had reached sexual maturity in the first year of their life, with the smallest length of 25 mm and 28 mm, respectively. The mean, standard length of the body was 50 mm. The spawning season was spread over four months from late-April to mid-August. Females laid eggs in portions, and the absolute and relative fecundity was 1372 and 1691, respectively. Stone moroko males were ready to spawn for a longer time period than females. The present study shows greater reproductive potential of stone moroko population in the central Europe than the native population, suggesting its successful colonisation in the new habitat.

DNA methylation stability in fish spermatozoa upon external constraint: Impact of fish hormonal stimulation and sperm cryopreservation.

Highly differentiated mature spermatozoa carry not only genetic but also epigenetic information that is to be transmitted to the embryo. DNA methylation is one epigenetic actor associated with sperm nucleus compaction, gene silencing, and prepatterning of embryonic gene expression. Therefore, the stability of this mark toward reproductive biotechnologies is a major issue in animal production. The present work explored the impact of hormonal induction of spermiation and sperm cryopreservation in two cyprinids, the goldfish (Carassius auratus) and the zebrafish (Danio rerio), using LUminometric Methylation Assay (LUMA). We showed that while goldfish hormonal treatment did increase sperm production, it did not alter global DNA methylation of spermatozoa. Different sperm samples repeatedly collected from the same males for 2 months also showed the same global DNA methylation level. Similarly, global DNA methylation was not affected after cryopreservation of goldfish spermatozoa with methanol, whereas less efficient cryoprotectants (dimethylsulfoxide and 1,2-propanediol) decreased DNA methylation. In contrast, cryopreservation of zebrafish spermatozoa with methanol induced a slight, but significant, increase in global DNA methylation. In the less compact nuclei, that is, goldfish fin somatic cells, cryopreservation did not change global DNA methylation regardless of the choice of cryoprotectant. To conclude, global DNA methylation is a robust parameter with respect to biotechnologies such as hormonal induction of spermiation and sperm cryopreservation, but it can be altered when the best sperm manipulation conditions are not met.

Effect of copper nanoparticles and ions on spermatozoa motility of sea trout (Salmo trutta m. Trutta L.).

Nanoproducts are being increasingly used in various industrial products, leading to a greater risk of water pollution through their discharge into environment as production byproducts. Increased levels of environmental pollution with nanoproducts pose a threat to all living organisms. Nanopollutants may have toxic effects on gametes and fertilization process in species with external fertilization, thereby reducing effectiveness of reproduction or greatly impairing it. The objective of the present study was to determine the effect of copper nanoparticles (Cu and CuO) and copper ions (CuSO4·5H2O) on the spermatozoa motility of sea trout and compare their harmful effects. Copper nanoparticles (NPs) of primary particle size <100 nm and CuO NPs of particle size <50 nm as well as a solution of CuSO4·5H2O were used for the study. Concentration of the products tested (CuNPs, CuONPs, and CuSO4·5H2O) was expressed as mg Cu L-1 in the media regardless of dissolution and aggregation. A suspension with concentration up to 500 mg L-1 in an aquatic environment was tested. Spermatozoa motility under direct contact with copper at 10 s post activation was evaluated using an automated system (CASA). Copper products in an aqueous environment primarily cause a reduction of spermatozoa velocity (VCL). A significant reduction of velocity in CuSO4·5H2O salt solution occurred at concentration starting from 8 mg L-1, whereas in solutions containing CuNPs and CuONPs this process is observed from 50 mg L-1. Reduction of percentage of motile spermatozoa occurred in CuNPs from a concentration of 125 mg L-1, while in CuONPs and CuSO4·5H2O, the effect began at concentration from 250 mg L-1. In a solution of CuSO4·5H2O at a concentration of 500 mg L-1, strong agglutination of sperm cells occurred and movement ceased (immobilization IC100). Exposure to CuONPs decreased the motility rate to below IC50, while for CuNPs, the motility rate reached 80%. Changes in linearity (LIN) and amplitude of head displacement (ALH) differed according to Cu products. Cu as CuSO4·5H2O and CuONPs impaired the LIN, while CuNPs lowered ALH. Duration of motility increased at low CuSO4·5H2O concentration. The effect of Cu ion on sea trout spermatozoa motility was more harmful than that of copper nanoparticles. The various copper products used affected motility parameters differently. Contamination of aqueous environment with copper primarily caused reduction in male gamete velocity, which may impair reproduction. Various copper products had different effects on LIN, ALH, and motility duration, which can also affect fertilization.

Inhibitory effect of K+ ions and influence of other ions and osmolality on the spermatozoa motility of European burbot (Lota lota L.).

BACKGROUND: In fish with external fertilization, two main start-up mechanisms of the path that blocks or activates the spermatozoan motility apparatus are known. The main factor managing the path is osmolality or potassium ion. In burbot from the European and North American population, contradictory findings regarding the factors influencing the onset of spermatozoa motility were reported. The objective of the current study was to determine the effect of potassium and osmolality on the spermatozoa activation of European burbot, Lota lota (Actinopterygii, Gadiformes, Lotidae). Moreover, the influence of pH, as well as sodium ion concentrations on spermatozoa motility was investigated. Seven parameters characterising motility were traced by means of computer-assisted sperm analysis (CASA). PRINCIPAL FINDINGS: The spermatozoa of European burbot are K+ ion-sensitive. A 6-mM KCl solution significantly decreased motility, and above 12-mM (50 mOsm kg-1) totally ceased spermatozoa movement. Sucrose and Na+ solutions inhibited spermatozoa movement only at concentrations > 450-480 mOsm kg-1. Greater differences in the percentage of motile sperm between individuals were noted in solutions containing high concentrations of chemicals triggering sperm motility. The optimum osmolality for spermatozoa motility is in the range of 100-200 mOsm kg-1. The burbot spermatozoa were motile over a wide range of pH values with the best activation at pH 9. CONCLUSION: It was demonstrated that the spermatozoa of European burbot are inhibited by K+ ions similarly as in North American burbot. Other electrolyte and non-electrolyte solutions inhibit spermatozoa movement only if their osmolality is greater than that of the physiological osmolality of seminal plasma. The data provided on basic knowledge of burbot spermatozoa allow to ensure appropriate conditions during artificial reproduction and scientific research.

Spermatozoa Motility Parameters of the Pumpkinseed (Lepomis gibbosus) in a Standardized Solution and Ambient Water.

The sunfish are classified as an invasive species after their introduction into a new habitat. Milt quality and spermatozoa movement in several sunfish were described. The aim of this study was to determine the basic milt characteristics and spermatozoa motility parameters in pumpkinseed males after using ambient water and a standardized solution to examine the parameters in mimicked natural conditions and in controlled conditions allowing interspecies data comparisons. Nine spermatozoa motility parameters were traced using computer-assisted sperm analysis (CASA) after activation with 30 mM NaCl and ambient water at 20°C. Spermatozoa motility parameters on activation, except for ALH, differed after using saline solution and ambient water. The CASA parameters were higher in the first medium. On the other hand, spermatozoa motility was high in both media, with an average of 89.6 and 85.2% in saline solution and ambient water, respectively. In the media, initial spermatozoa motility was characterized by a mean curvilinear velocity of 122.4 and 108.5 µm s-1, and a linearity of 79.7 and 67.1%, respectively. The active motility phase in the saline solution was approx. 8 min was longer than that measured in ambient water (mean 3 min). The pumpkinseed spermatozoa reveal good motility parameters, especially the high percentage of motile sperm, linearity, long duration of spermatozoa movement, as well as good velocity. It can be assumed that apart from reproductive strategy, spermatozoa motility parameters are valuable features of the male reproductive biology which may contribute to reproductive success.

Spermatozoa concentration influences cryopreservation success in sea trout (Salmo trutta m. trutta L.).

Variation among individuals is substantial for spermatozoa concentration in fresh milt in sea trout (Salmo trutta m. trutta L.). The objective of the present study was to examine effects of spermatozoa concentration in this species on subsequent cryopreservation success. Milt with high spermatozoa concentration was diluted with seminal plasma to obtain concentrations ranging between 6 and 24 × 10(9) mL(-1) with steps of 2 × 10(9) mL(-1). Diluted milts were cryopreserved in 0.25-mL straws with extender (0.3 M glucose) containing 10% methanol and 10 % (vol/vol) supplement of hen egg yolk. The dilution ratio was 1:3 (milt:cryomedium). Cryopreservation efficacies were assessed according to evaluation of motility of frozen/thawed spermatozoa and quantification of fertilizing ability. Percentage of motility of frozen/thawed spermatozoa was influenced by spermatozoa concentration in the cryomedium (P < 0.05). The highest motility was observed in samples with 3.0 to 4.0 × 10(9) spermatozoa per mL of cryomedium, which corresponds to 12 to 16 × 10(9) spermatozoa per mL in fresh milt. Higher sperm concentrations and lower sperm concentrations in cryomedium reduced the effectiveness of cryopreservation when compared with the optimum. Cryopreservation success measured according to fertilization rate was in agreement with results for motility of frozen/thawed spermatozoa, but the optimum could not be determined with statistical precision because of differences in fertilization rate among individual donor males. However, a significant positive correlation was found between postthaw motility and fertilization rate and between cryopreserved spermatozoa velocity and fertilization rate (P < 0.05). In sea trout, cryopreservation efficiency is influenced by spermatozoa concentration in cryomedium. Individual adjustment of the dilution ratio, based on initial spermatozoa density, is recommended in the freezing protocol. Maximum cryoresistance of the cell was obtained when spermatozoa concentration in cryomedium ranged from 3.0 to 4.0 × 10(9) mL(-1).

Effect of pH and cation concentrations on spermatozoan motility of sea trout (Salmo trutta m. trutta L.).

Environmental conditions during external fertilization in fish have a significant effect on spermatozoan motility (MOT) and fertilization ability. Even in the same family of fish, spermatozoa might differ in sensitivity to ions present in the external medium. Elucidation of such differences within a species would help to understand spermatozoan biology and to determine external conditions that would optimize spermatozoan MOT and successful fertilization. Objectives of the current study were to determine the effects of pH and of sodium, potassium, and calcium ion concentrations in the activation solution on sea trout spermatozoan MOT. Six parameters characterizing MOT (MOT, curvilinear velocity [VCL], linearity, amplitude of lateral head displacement, beat cross frequency, and duration of MOT) in spermatozoa activated in prepared buffers were traced by computer-assisted sperm analysis. Sea trout spermatozoa were motile over a wide range of pH values, and increasing pH did influence MOT, VCL, linearity, amplitude of lateral head displacement, and MOT duration. The optimum pH for sperm MOT was established at approximately 10. Increasing K(+) ion concentration within the observed range caused a decrease in MOT and VCL. Spermatozoan movement ceased at 8 mM KCl concentrations. In Ca(2+) buffers, sperm were motile within the range of 0 to 70 mM CaCl(2) concentration; although beyond 8 mM concentration, VCL and MOT gradually declined. Spermatozoan aggregation was observed at the highest ion concentrations tested. Increasing CaCl(2) concentration affected MOT pattern from initiation to termination of spermatozoan movement in a similar manner as changes associated with increasing pH. At concentrations of CaCl(2) higher than 0.5 mM and in buffers with pH values 10 to 11, movement of spermatozoa was characterized by high initial linearity followed by its gradual reduction. In contrast to the effects of KCl and CaCl(2), increasing NaCl concentration up to 90 mM Na(+) concentration prolonged the duration of spermatozoan movement and, up to 60 mM Na(+) concentration, slightly increased sperm velocity as well. Above the concentration of 90 mM NaCl, these parameters decreased; and at 240 mM of Na(+), spermatozoa did not activate.

Body size versus gonad maturation form in under-yearling precocious males of the sea trout (Salmo trutta m. trutta L.).

The study was aimed at analysing body size in relation to form of gonad maturation (amount of mature germ cells) in 329 under-yearling sea trout males. The fish, aged 7 months, were caught in late October-early November in 3 streams located in north-western Poland. Each stream supported fish belonging to a different sib group. Standard histological techniques and a computer image analysis programme were used to detect the class of gonad maturation and percentage of the gonad area occupied by tubules with active spermatogenesis. Gonad maturation forms were distinguished based on the latter criteria. Gonads with developing germ cells occupying less than 90% of gonad area were classified as incomplete forms of gonad maturation, others as complete maturation forms. In each sib-groups analysed, even the smallest individual were already precocious, their gonads being incompletely mature. The smallest maturing male measured 7.1 cm in length. The average size of an incompletely maturing individual was slightly smaller than that of the completely mature one but the difference lacked statistical significance (P > 0.05). The sib-group of smaller fish contained less precocious, and the gonads of the more precocious were incompletely mature, compared to the sib-group of larger fish (P < 0.001). It seems that the incomplete form of gonad maturation (defected maturation) occurs at a smaller critical fish size than the complete gonad maturation form. Incomplete maturation is more frequent smaller individuals and possibly in among slow-growing groups of fish.

Histology of salmonid testes during maturation.

The commonly applied classification systems of fish gonad maturity divide the maturation process into certain stages. However, the scales do not entirely reflect the continuity of the maturation process. Based on light microscope observations, the paper describes a comprehensive pattern of testicular transformations during maturation. The study was carried out on precocious underyearling and 1-year-old males of sea trout (Salmo trutta m. trutta L.), 1-year-old males of salmon (Salmo salar L.), and males of brown trout (Salmo trutta m. fario L.) aged from 7 months to 4 years. A total of 821 gonads collected during all seasons of the year were examined. The fish were fixed in Bouin's fluid. Histological slides of the mid-part of the gonad were made using the standard paraffin technique. The 3-6 microm sections were stained with Heidenhain haematoxylin. Histological changes of testes during maturation were similar in the three species studied. Immature and resting gonads contained type A spermatogonia in lobules only. The appearance of cystic structures containing type B spermatogonia in the lobules signalled the beginning of the sexual cycle in male gonads. Type B spermatogonia underwent synchronous mitotic divisions resulting in an increase in the total number of spermatogonia. As the spermatogenesis continued, the gonads showed a gradual increase in the number of cysts containing cells at all the spermatogenetic stages: type B spermatogonia, primary and secondary spermatocytes, spermatids, and spermatozoa. The well-formed spermatozoa were released to the lobule lumen once the Sertoli cells and spermatozoa connections broke up and the cyst disappeared. This was a continuous process observed throughout the spawning season. The spermatozoa were moved to the efferent duct. While some of the germ cells were completing spermatogenesis, the lobules contained less and less cysts with type B spermatogonia, primary and secondary spermatocytes, and spermatids; eventually all the cells completed spermatogenesis. At the end of maturation, vacuoles, up to 18.9 microm in final diameter (brown trout), appeared in the Sertoli cells. The vacuoles were visible in the lobule wall epithelium for a prolonged period of time. In most salmonid individuals examined, the reproductive cycles were observed to overlap. In some fish, the preparation for another cycle began very early, i.e., at the and of preceding spermatogenesis, which had not been observed before. Gonad maturation in some males was incomplete.