Amplitude Of Lateral Head Research Articles

Effect of Skim Nanoparticles on the Motility and Kinematics of Simmental Cattle Frozen Semen

Nanotechnology has positively impacted the development of extenders used in the cryopreservation of spermatozoa in livestock. The inclusion of nano skim in the extender is expected to preserve semen quality after thawing. This study aimed to determine the optimal concentration of nano skim. Various concentrations of nano skim (0%, 1.66%, 3.33%, 5%, 6.66%, 8.33%, and 10%) with the addition of 10% nano egg yolk in each extender were compared. Fresh semen samples were collected from three three-year-old Simmental bulls using an artificial vagina. The study parameters included motility (total motility, progressive motility) and kinematics (velocity curve linear (VCL), velocity straight linear (VSL), velocity average pathway (VAP), amplitude lateral head (ALH), beat cross frequency (BCF), straightness (STR), linearity (LIN), and wobble (WOB). A completely randomized design was employed, and data were analyzed using analysis of variance (ANOVA). Fresh semen dilution results indicated that only the control group (0%) and the nano skim group with concentrations of 6.66% and 8.33% were eligible for further processing into frozen semen. The results showed no significant difference between the three treatment groups on sperm motility and kinematics of Simmental cattle after thawing (p0.05). The control group exhibited the highest percentage of total motility, VCL, VAP, and ALH values, while the 6.66% nano skim group outperformed the 8.33% nano skim group. The highest percentage of progressive motility and VSL value was observed in the 6.66% nano-skim group compared to the 8.33% nano-skim and control groups. The highest values of BCF, WOB, LIN, and STR was observed in the nano-skim group at 8.33% compared to the nano-skim group at 6.66% and control. The study concluded that 6.66% nano skim was the optimal concentration to maintain the quality of frozen semen of Simmental cattle.

Quercetin in semen extender improves frozen-thawed spermatozoa quality and in-vivo fertility in crossbred Kamori goats.

This study investigated the antioxidant effect of quercetin-treated semen on frozen-thawed spermatozoa quality and in-vivo fertility in crossbred Kamori goats. In total, 32 ejaculates from four fertile bucks were diluted in Tris-based egg yolk extender with varying levels of quercetin (0, 1, 5, 10, and 15 μM). Qualified semen samples were pooled and frozen in French straws. The results revealed that the addition of quercetin in the semen extender increased (p < 0.05) frozen-thawed sperm total motility (TM), progressive motility (PM), rapid velocity (RV), average path velocity (VAP), straight line velocity (VSL), curvilinear velocity (VCL), and amplitude of lateral head (ALH) displacement in contrast to the control group. Quercetin supplementation had no effect on beat cross frequency (BCF), straightness (STR), and linearity (LIN) (p > 0.05). Quercetin showed significantly higher (p < 0.05) plasma membrane and acrosome integrity and viability (p < 0.05) of spermatozoa in contrast to the control group. Quercetin in the semen extender significantly increased (p < 0.05) superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), ascorbate peroxidase (APX), and total antioxidant capacity (TAC) levels while reduced (p < 0.05) the contents of total oxidant status (TOS) and malondialdehyde (MDA), which were in contrast to the control group. Ultrasound results revealed that 24 out of 30 (80%) goats were found pregnant when semen was treated with 5 μM quercetin while the control group showed 18 out of 30 (60%) animals were pregnant. Thus, the study concluded that 5 μM quercetin-treated semen was found to be efficient, showed increased antioxidant status, and reduced oxidant production, leading to improved spermatozoa quality and in-vivo fertility in goats.

Post-thaw sperm morphokinetics and mitochondrial genes profile of buck semen supplemented with selenium in two different forms

Background The quality characteristics of semen is a determinant factor for improving fertility of rabbit. Objectives The current investigation was done to evaluate post-thaw sperm morphokinetics and mitochondrial genes expression profile of buck semen supplemented with selenium in two different forms. Materials and methods Sexually mature bucks (n=16) that aged three months on average were used for collecting two ejaculates which were evaluated using computer assisted semen analysis (CASA) program. The semen samples of all experimental animals which recorded progressive motility greater than or equal to 70% were pooled for cryopreservation. The pooled semen was divided into three groups that were supplemented with selenium nanoparticles and selenium in normal form in addition to the control group. Morphological characteristics as well as CASA parameters were assessed after freezing for 1 week. Moreover, enzymatic activity assays were performed to measure the antioxidant capacity of cryopreserved buck semen. Transcriptional profile of mitochondrial activity and antioxidant defense regulating genes was conducted using quantitative real-time polymerase chain reaction (PCR). Results and conclusion The addition of selenium in normal and Nano forms has significantly (P ≤0.05) enhanced some of CASA parameters such as DCL (µm), DAP (µm), DSL (µm), VCL (µm/s), VAP (µm/s), Amplitude of lateral head (ALH) (µ), BCF (Hz), and VSL (µm/s) during prefreezing period compared with control group. Moreover, the data presented in the present study indicated a significant (P ≤0.05) improvement of post-thaw total and progressive sperm motility in the two groups supplemented with normal and Nano compared with control group. The post-thaw level of total antioxidant capacity (TAC) and percentage of live sperm were higher in the two groups supplemented with selenium and Nano selenium than the control group. The expression profile of candidate genes regulating mitochondrial activity (ATP Synthase F1 Subunit Alpha (ATP5A1), NADH dehydrogenase subunit1 (ND1), NADH dehydrogenase subunit2 (ND2), and Carnitine palmitoyltransferase (CPT2)) was increased significantly (P ≤0.05) in semen supplemented with selenium in normal and Nano forms compared with the control group. In conclusion, the data of this investigation demonstrated enhancement of CASA parameters during pre-freezing post thaw total and progressive sperm motility in the two groups supplemented with normal and nano selenium. The proportion of sperm viability and the level of total antioxidant capacity were enhanced in the two groups supplemented with selenium which was coupled with up-regulation of mitochondrial transcripts. It seems the both two forms shared the same mechanism on improving post-thaw physical and molecular qualities of rabbit sperm.

Analysis of Motility Characteristic of Pesisir Bulls Sexed Semen with Different Pre-Freezing Method Based on Computer Assisted Sperm Analyzer (CASA)

This study aimed to find the best combination between pre-freezing method and layer of post sexing on semen quality of Pesisir Bulls. This study used 2 Pesisir Bulls. The semen was evaluated using Computer Assisted Sperm Analyzer (CASA). This study used factorial randomized block design with 2 factors. Factor A was sexed semen layer of BSA column with 2 level, whereas Factor B was pre-freezing method with 3 level. Analysis of Variance (ANOVA) were used in this study for data Analysis. According to the analysis of variance, there is no interaction of the combination of two factors on each paramters. Pre-freezing method showed highly statistically significant effect (P&lt;0.01) on the quality of motility on upper layer and bottom layer which was about (52.86-72.93%; 52.32-65.50%) and about (40.17-56.99%; 40.33-53.18%) on progressive motility. The value of Distance Curve Line on (39.15-43.24 μm; 49.25-53.86 μm), Distance Average Path (22.38-24.60 μm; 26.52-29.94 μm), Distance Straight Line (17.32-19.07 μm; 20.93-24.25 μm). The value of Velocity Curve Line (88.67- 97.50 μm/s; 109.85-117.90 μm/s), Velocity Average Path (50.98- 55.63 μm/s; 59.32- 64.69μm/s), Velocity Straight Line (34.45-42.37 μm/s; 46.80-50.62 μm/s), Beat Cross Frequancy (23.91-25.17 Hz; 25.63-27.35 Hz), Straightness (0.75-0.77; 0.77-0.80), and Wobble (0.56-0.57; 0.53-0.55), highly statistically significant (P&lt;0.01) on post sexing layers. Meanwhile, the value of Amplitude of Lateral Head (4.24-4.94 μm; 4.25-4.44 μm) showed highly significant effect (P&lt;0.01) on factor A and factor B treatment, and value Linearity was about (0.42-0.44; 0.43-0.44) showed not significant effect (P&gt;0,05) on every factor. According to the study, it can be concluded that the best treatment was treatment modified procedure layer by space the straw 16 cm every layers and The quality of post sexing X- and Y- spermatozoa motility effect on motility characteristic such as DCL, DAP, DSL, VCL, VAP, VSL, STR, WOB, ALH and BCF.

Semen Cryopreservation Quality and Sperm Kinematics of Saanen Goats Using Different Diluents

The success of artificial insemination (AI) in small ruminants, especially goats, depends on the quality of frozen semen. Therefore, the current study aimed to determine the quality of various diluents, including tris-egg yolk, AndroMed®, and OviXcell®, on semen quality. The fresh semen samples from three male Saanen goats aged 1.5-2 years were collected and the mean individual motility of samples was recorded at 70%. The cryopreservation quality of the semen was evaluated based on motility, viability, abnormality, and total sperm motility (TSM) indexes. The present laboratory experiment was performed with 3 treatments and 10 repetitions. The treatments in this study were T0 (tris-egg yolk), T1 (AndroMed®), and T2 (OviXcell®). The results showed no significant difference in the parameters of motility, viability, abnormality, and TSM among the treatment group. The kinematic parameters’ average path length, velocity curved linear, and linearity showed a significant difference in all treatment groups. However, there were no significant differences among the three groups in terms of motility, progressiveness, distance curved line, distance straight line, average velocity path, velocity straight line, straightness, amplitude lateral head, beat cross frequency, and wobble kinematic parameters. Motility was higher in T2 than in T0 and T1, viability was higher in T1 than in T0 and T2, and abnormality was lower in T1 than in T0 and T2. In conclusion, the use of various diluents, such as tris-egg yolk, AndroMed®, and OviXcell®, can maintain the quality of frozen spermatozoa for over 24 hours, including motility, viability, abnormality, and TSM. Kinematic parameters obtained using CASA IVOS II can provide relevant information for various parameters using these diluents.

Evaluation of equine semen frozen in extenders free of animal proteins

Various countries are reticent regarding importation of semen that contains egg-yolk, limiting its commercialization. Egg-yolk composition is complex, varying with the diet of the chicken and egg-yolk granules are similar in size to sperm, interfering with analysis. Therefore, development of a chemically defined cryopreservation extender that maintains seminal parameters is relevant. Twelve ejaculates from 4 stallions of proven fertility (n= 4; r=3) were diluted in one of 5 extenders: 1) EDTA-glucose with 11% lactose, Equex, 20% egg-yolk and 5% dimethylformamide (EY); 2) casein salt and cyclodextrin cholesterol extender (CE); 3) CE with 3% dimethylformamide (CE-3); 4) OptiXcell (OP); 5) BioXcell (BIO). Cryopreservation was carried out in a Cryogen, Neovet automatic freezing machine: -0.5°C/minute to 5°C, 180 minutes equilibration, then -5°C/minute to -20°C and -20°C/minute to -100°C, then plunging in N2. Thawing was carried out at 37°C for 1 minute and sperm kinematic parameters (CASA, AndroVision, Minitube), viability and acrosome status (FITC-PNA-PI stain), membrane lipoperoxidation (BODIPY stain) and DNA fragmentation (SCD) were evaluated. Data were analyzed using ANOVA or Friedman test and results are reported as mean ± SD. Higher (P<0.05) percentages of total and progressively motile sperm were observed in samplescryopreserved in EY (29.25 ± 11 and 21.17 ± 10), CE-3 (30.4 ± 15 and 22 ± 12) and OP (23.1 ± 14 and 15.5 ± 10) compared to those in CE (4.75 ± 3 and 1.58 ± 2) and BIO (8 ± 4 and 3.25 ± 3). Straight-line, curvilinear and average-path velocity parameters, amplitude of lateral head and beat-cross frequency were significantly higher in samples frozen with EY, CE-3 and OP compared to CE and BIO media. Straight-line and average-path velocities were also significantly higher in OP compared to EY medium. Live acrosome-intact sperm were significantly higher in EY (45 ± 14) compared to the rest of the extenders; with CE-3 (25 ± 10) being significantly higher than CE (8 ± 5), OP (9 ± 5) and BIO (13 ± 5). Live, non-peroxidized sperm were significantly higher in EY (47 ± 15) compared to the other extenders; with CE-3 (29 ± 15) being significantly higher than CE (14±10), OP (21 ± 15) and BIO (23 ± 14). No significant differences were observed in the percentages of sperm with intact DNA in any of the extenders: EY (95 ± 4); CE (95 ± 2); CE-3 (93 ± 8); OP (94 ± 7) and BIO (94 ± 5). To conclude, the casein salt and cyclodextrin cholesterol extender with 3% dimethylformamide, without egg-yolk could be a suitable alternative for extenders with animal proteins. Further studies are ongoing to evaluate in vivo fertility of ejaculates cryopreserved in this extender.

Effect of modified CEP-3 diluents with aqueous soybean extract on liquid semen quality in Ongole crossbred bull.

Egg yolk (EY) is commonly used as an extracellular cryoprotectant in semen diluents but has some negative effects. Therefore, this study aimed to assess the potential of lecithin derived from plants, such as soybeans, as an alternative extracellular cryoprotectant and to characterize liquid semen quality of Ongole crossbred bulls using a modified caudal epididymis plasma-3 [CEP-3 (m)] as a base diluent and aqueous soybean extract (ASE). A bull with progressive motility (PM) of fresh semen >70% was used. Two soybean extracts were also used, namely, ASE 1 and ASE 2, obtained by extraction procedures 1 and 2, respectively. The study was conducted using an experimental design with 11 treatments and ten replications, with diluents comprising different levels of ASE 1 and ASE 2, as well as a positive control with 10% EY. The parameters measured were motility (M) and its kinetic parameters, including PM, M, velocity curve linear, velocity straight linear, velocity average pathway, linearity, straightness, wobble, amplitude lateral head beat cross frequency, and hyperactivity using computer-assisted sperm analysis, viability, and spermatozoa abnormalities. The CEP-3(m) diluent formula and ASE 1 at a 30% level maintained the PM of spermatozoa up to day 5 (40.7% ± 16.1%) of cold storage. Meanwhile, the CEP-3(m) diluent formula and ASE 2 could only maintain PM >40% until day 3 (42.1% ± 13.5%) of cold storage at a 30% level. The CEP-3(m) diluent and ASE 1 at a level of 25%-30% supported spermatozoa life (viability) up to day 5 with a value >80% (81.8 ± 3.5; 86.4 ± 2.6). The abnormality value of spermatozoa in various diluents during cold storage on days 0-5 was below 20%. Soybean extracts 1 and 2 can substitute EYs as extracellular cryoprotectants in modified CEP-3 basic diluents. Soybean extract 1 can support the life of spermatozoa up to day 5 but may cause the viscosity and movement of spermatozoa to be hyperactive. Soybean extract 2 can support the life of spermatozoa up to the 3rd day of cold storage and produces progressive (non-rotating) movement patterns. Further, research is recommended with higher levels of ASE 2.

Exogenous GSH Supplementation to Raw Semen Alters Sperm Kinematic Parameters in Infertile Patients.

Glutathione is an important antioxidant found in all mammalian cells. Sperm motility is positively correlated with seminal reduced glutathione (GSH) levels, and infertile men are known to have lower GSH levels. Studies on GSH supplementation in improving sperm functions in infertility patients are limited. Here, we re-investigate the effect of exogenous GSH supplementation on human sperm motility and kinematic parameters. Residual semen samples from 71 infertility patients who came for routine semen analysis for infertility assessment were studied. Liquefied raw semen was supplemented with GSH (0-10 mM) for 1 h. The untreated sample was the blank control. Only a 5 mM concentration was tested in all 71 samples. After two washes, the sperm was incubated and then analyzed for sperm motility and kinematic parameters by computer-assisted semen analysis (CASA), followed by adenosine triphosphate (ATP), reactive oxygen species (ROS) levels, free thiols, and DNA damage analyses. At 2 hrs post-treatment, GSH supplementation significantly altered many of the kinematics, compared to the control. Straight line velocity (VSL) (p = 0.0459), curvilinear velocity (VCL) (p < 0.0001), average path velocity (VAP) (p < 0.0001), and lateral head amplitude (ALH) (p < 0.0001) were decreased, whereas straightness (STR) (p = 0.0003), linearity (LIN) (p = 0.0008), and beat cross frequency (BCF) (p = 0.0291) were increased in 5 mM group. Wobble (WOB) (p = 0.4917), motility (MOT) (p = 0.9574), and progressive motility (PROG) (p = 0.5657) were unchanged. ATP level was significantly increased in the 5 mM group (p < 0.05). It is concluded that exogenous GSH supplementation does alter sperm kinematics in humans. These altered kinematic parameters together with increased energy (ATP) may have a positive role in influencing the success rates of ART procedures.

Effect of Detergents Based on Sodium Dodecyl Sulfate on Functional Metrics of Frozen-Thawed Collared Peccary (Pecari tajacu) Semen.

We evaluated the effects of detergents based on sodium dodecyl sulfoxide (SDS) on the functional parameters of collared peccary frozen-thawed sperm. Semen aliquots from ten individuals were diluted in a Tris-egg yolk-glycerol extender alone or with 0.5% Equex STM® paste or SDS (at 0.1%, 0.3% or 0.5% (v/v) concentration). Samples were fast frozen in liquid nitrogen with a post-thaw evaluation of motility, membrane functionality and integrity, mitochondrial activity, sperm binding ability and thermal resistance. The treatments without SDS (41.8 ± 3.5%) and those containing Equex (41.8 ± 4.4%) or 0.1% SDS (41.2 ± 5.5%) provided greater sperm motility (p < 0.05) than those containing SDS 0.3% (30.5 ± 4.7%) and 0.5% (31.2 ± 6.3%). Immediately after thawing, only treatments containing 0.1% SDS effectively preserved sperm straightness (STR) when compared to the negative control. All treatments preserved the amplitude of lateral head (ALH) and straightness (STR) during a thermal resistance test (p > 0.05), but SDS 0.5% impaired the membrane functionality and mitochondrial activity after thawing (p < 0.05). All treatments provided a similar recovery of sperm binding ability after thawing (p < 0.05). Our results showed that the addition of 0.1% SDS to the Tris-yolk-glycerol extender optimized the freeze-thaw recovery of peccary semen.

Semen parameters are seriously affected in acephalic spermatozoa syndrome

BackgroundPrevious studies have reported that some patients with headless spermatozoa have poor semen quality, but there has been no published systematic analysis of semen quality in patients with different proportions of headless spermatozoa in semen. We aimed to explore the association of acephalic spermatozoa syndrome and semen quality in men with distinct proportions of headless spermatozoa.Material and methodsSemen parameter values in patients for whom headless spermatozoa were found in the ejaculates was studied and compared to that of 413 age-matched prenatal examination patients. All semen samples were analyzed following the same methodology in a single laboratory.ResultsAll semen parameter values except semen volume were negatively (P < 0.05) correlated with the proportion of headless spermatozoa. The semen samples were divided into four groups on the basis of the proportion of headless spermatozoa (PHS) as follows: 0 < PHS ≤ 5% (n = 172, Group A1); 5 < PHS ≤ 10% (n = 76, Group A2); 10 < PHS ≤ 20% (n = 71, Group B); and PHS > 20% (n = 71, Group C). In Group A1, only one semen parameter value (progressive motility) was lower than those of the control group, but in Group A2, this increased to five (sperm vitality, normal sperm morphology, sperm motility, VCL (curvilinear velocity) and ALH (amplitude of lateral head displacement)). Worse still, all semen parameter values were significantly lower in Group B and Group C than in the control group (P < 0.05).ConclusionsSemen samples containing headless spermatozoa tend to have lower quality than samples without headless spermatozoa. Increases in the proportion of headless spermatozoa in semen are associated with decreased semen quality. We suggest that headless spermatozoa should be seriously assessed and accurately counted in semen analysis, especially for ejaculate in which the proportion of headless spermatozoa exceeds 5%.

Aroclor 1254 impairs sperm quality, fertilization ability, and embryo development of rainbow trout (Oncorhynchus mykiss).

The polychlorinated biphenyls (PCBs) in aquatic environment adversely affect non-target organisms, including fish. Especially, the male reproduction and next generation can be damaged through high exposure to these pollutants. Hence, the sperm cells were exposed to sublethal concentrations of Aroclor 1254 (0, 1, 5, 10, or 25mg/l) for 4h. The sperm quality parameters were analyzed by SCA (Sperm Class Analyzer). The fertility, eyeing, and hatching rates were determined as gamete markers. Lipid peroxidation (malondialdehyde-MDA), glutathione (GSH), and antioxidant enzymes [superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT)] were measured for determination of oxidative stress. Our results showed that Aroclor 1254 negatively affected the motility rate and duration, fertilization rate, embryogenesis, and hatching and also triggered antioxidant defense mechanisms at the highest concentration (25mg L-1). Furthermore, linear speed (VSL), linearity index (LIN), and amplitude lateral head (ALH) were significantly changed after exposure to 25mg L-1, and the lowest concentrations (1 and 10mg L-1) did not significantly affect the motility and fertilizing capacity. The embryogenesis and hatching were significantly affected by sperm exposure to 1, 10, and 25mg L-1 of Aroclor 1254. Consequently, Aroclor 1254 causes potential hazards in male germ cells, and the exposure of sperm cells to pollutants can adversely affect next generation of wild populations.

P-022 Relationship between sperm movement characteristics assessed by a sperm motility analysis system (SMAS) and normal fertilization rates in vitro

Abstract Study question Is there a relationship between movement characteristics of human sperm assessed by SMAS and normal fertilization rates in vitro? Summary answer The curvilinear path wobble of motile sperm before insemination is predictive of normal fertilization rates after in vitro fertilization (IVF). What is known already Assessment of semen parameters is very important in both the diagnosis of male infertility and the selection of methods for treatment with conventional-IVF (C-IVF) or intracytoplasmic sperm injection (ICSI). Several studies have reported that objectively movement characteristics of motile sperm assessed by SMAS are related to the functional capacity of human sperm. However, there are no established the objective indicators and criteria for sperm motility characteristics assessed by SMAS. In this study, we investigated the utility of the movement characteristics of motile sperm measured by SMAS as an indicator of normal fertilization rates. Study design, size, duration Retrospective experimental trial conducted in a single IVF center between January 2018 and February 2020. Sperm movement characteristic analysis was performed using the SMAS in 300 IVF cycles using sperm from 270 men. At least three metaphase II oocytes were inseminated in C-IVF. The SMAS estimates were measured in semen and before insemination. This study was approved by the Ethics Committee of the Institutional Review Board (IRB) of Kinutani Women’s Clinic, Hiroshima, Japan. Participants/materials, setting, methods Motile sperm from at least 3 different fields were observed in each sample and their parameters were examined. First, we investigated the correlation between the normal fertilization rates and sperm motility characteristics. Next, the group comprising 300 cycles was classified into a group of a ≥ 60% normal fertilization (group 1: 229 cycles) and a group with &amp;lt;60% normal fertilization (group 2: 71 cycles) to assess the sperm characteristics described below. Main results and the role of chance The following indicators were evaluated: amplitude lateral head displacement (ALH), best cross frequency (BCF), curvilinear velocity (VCL), straight-line velocity (VSL), average path velocity (VAP), linearity (LIN=VSL/VCL), straightness (STR=VSL/VAP), and curvilinear path wobble (WOB=VAP/VCL).There were significant correlations between normal fertilization rates and semen parameters, including VCL (r = 0.014; P &amp;lt; 0.05) and ALH (r = 0.177; P &amp;lt; 0.05). However, there were no significant correlations for other parameters. Moreover, there were no significant correlations between normal fertilization rates and sperm parameters before insemination. The SMAS estimates in semen and sperm before insemination were compared between group 1 and group 2. There were no significant differences in most baseline characteristics across groups, except for the number of normally fertilized oocytes. However, there were significant differences in sperm WOB before insemination between group 1 and group 2 (0.589 vs. 0.572; P &amp;lt; 0.05). Sperm motility before insemination was more curvilinear in group 1 than in group 2. The optimal cut-off value for WOB of motile sperm before insemination for predicting≥60% normal fertilization was 0.587 (AUC was 0.60; P &amp;lt; 0.05). These results were suggested that the WOB of sperm before insemination is useful as an indicator in determining the selection of methods for fertilization in vitro. Limitations, reasons for caution The main limitations of this study are its retrospective design and small sample size. A prospective study is necessary to further clarify the prediction criteria. Wider implications of the findings This study suggests that the WOB of motile sperm before insemination may be used as one of the indicators to predict good fertilization rates after C-IVF. Trial registration number N.A.

Comparison of sperm motility and density in four different goldfish (Carassius auratus) types

Different goldfish types play an important role both in ornamental fish farming and science. Considering its historical background, the goldfish is a suitable model animal for the study of artificial selection as well as for developmental biological studies. Sperm motility and cell density is an important parameter in determining sperm quality. The aim of our study was to examine the effects of different goldfish types on the sperm quality. Several sperm motility parameters (progressive motility (pMOT, %), straight line velocity (VSL, μm s-1), curvilinear velocity (VCL, μm s-1), linearity (LIN, %), amplitude lateral Head Displacement (ALH, μm), Beat Cross Frequency (BCF, Hz)) of four different goldfish types (Common goldfish-“wild type” N=5, Shubunkin N=4, Black Moor N=4, Oranda N=5) was compared during 60 hours (at 12-hour intervals) at refrigerated storage (4 °C). The variability of sperm density was also investigated in all types. A similar cell concentration was determined in the four goldfish variants (Common goldfish 2.01*1010 ± 3.46*109; Shubunkin 1.71*1010 ± 3.25*109; Black Moor 1.66*1010 ± 3.02*109; Oranda 1.56*1010 ± 5.83*109). Statistically significant difference between the 4 goldfish types in the motility parameters and cell density was not noted. However, a decreasing tendency in Black Moor sperm motility parameters (pMOT, VCL and VSL) was observable, as well as a reduced spermatozoa density in Oranda was also recorded. Our results can contribute to the improvement of the common hatchery propagation of goldfish. Future studies can add more evidence of the possible effects of artificial selection on the reproduction in different goldfish types.

Efficacy of soursop juice extender on rooster semen quality, oxidative activity and spermatozoa kinematics.

African medicinal plant like soursop (Annona muricate L.) within annonaceae are known for their biological, therapeutic, and pharmacological properties with little or no toxicity. The use of such plant requires good knowledge of the toxicity dosage, purity, suitable extraction solvent and adverse effects. The leaves, seeds, fruits, barks, and roots of African medicinal plants have been used for various nutraceuticals and functional effects according to African folk medicine. The aim of this study is to evaluate the semen quality, oxidative activity and spermatozoa kinematics of rooster semen in soursop juice extender. About 30 roosters were used for the in vitro analysis. Semen was collected twice a week for 2 weeks through dorsal-abdominal massage technique. The evaluation was done hourly until semen quality declined at the 5th-hour. The pooled semen was allotted to seven treatments of semen extenders as undiluted semen, dextrose saline, 10% soursop juice extender, 20% soursop juice extender, 30% soursop juice extender, 40% soursop juice extender, and 50% soursop juice extender for the study. The percentage motility, progressive motility, nonprogressive motility, curvilinear velocity, average path velocity, straight line velocity, linearity, straightness, amplitude of lateral head, beat cross frequency and wobble were analyzed using computer aided sperm analysis. Oxidative status (antioxidant activity and lipid peroxidation) was determined by assay. Result of rooster semen at room temperature and after 1-hour dilution showed that percentage motility, nonprogressive motility, and average path velocity were significantly (P < 0.05) reduced by different soursop juice extenders compared to undiluted semen. After 2-hour dilution of rooster semen, nonprogressive motility, average path velocity, curvilinear velocity, straight line velocity, wobble, liveability and amplitude of lateral head parameters were significantly (P < 0.05) increased by different soursop juice extenders compared to undiluted semen. Antioxidant activity and lipid peroxidation in both room temperature and after 5-hour dilution were affected by different soursop juice extenders in rooster semen. In conclusion, supplementation of soursop juices as an extender to rooster undiluted semen played an improvement role on spermatozoa fertility and oxidative status during processing or preserving ejaculates for insemination.

The Use of Animal's Body, Scrotal Temperature and Motion Monitoring in Evaluating Boar Semen Production Capacity.

Simple SummaryThe evaluation of male animals’ fertilizing capacity is based on laboratory-performed tests, as well as on the in vivo and in vitro reproductive outcome. Boar fertility is strongly associated with a pig farm’s financial status and income. The scientific community attempts to increase the predictive reliability of male fertility to ensure the profitability of the pig industry. In this direction, a different approach, using biomedical thermal and motion sensors, is utilized. This study investigates the correlation of collected data by biomedical sensors with variables well known as their predictable value semen quality. Strong correlations are revealed, encouraging the further application of biomedical methods in the assessment of boars’ fertilizing ability.Biomedical measurements by specialized technological equipment have been used in farm animals to collect information about nutrition, behavior and welfare. This study investigates the relation of semen quality (CASA analysis, viability, morphology, membrane biochemical activity and DNA fragmentation) with boar behavior during ejaculation. Sensors were placed on the boar’s body. Movement features were collected using an inertial measurement unit (IMU), comprising an accelerometer, a gyroscope and a magnetometer. Boar, scrotal and dummy temperatures were measured by an infrared (IR) camera and an IR thermometer, while the face salivation of the boar was recorded by a moisture meter (also based on IR technology). All signals and images were logged on a mobile device (smartphone or tablet) using a Bluetooth connection and then transferred wirelessly to the cloud. The data files were then processed using scripts in MATLAB 2021a (MathWorks, Natick, Massachusetts) to derive the necessary indices. Ninety-four ejaculates from five boars were analyzed in this study. The statistical analysis was performed in the Statistics and Machine Learning Toolbox of MATLAB 2021a using a linear mixed effects model. Significant and strong negative correlations (R2 > 0.5, p ≤ 0.05) were observed between boar, dummy and scrotal temperature with the progressive, rapid and slow movement of spermatozoa, VCL (curvilinear velocity), VSL (straight line velocity) and ALH (amplitude of lateral head displacement) kinematics. The volume of the ejaculate was correlated with the scrotal and dummy temperature. Dummy’s temperature was negatively correlated with BCF (beat/cross-frequency), viability and total time of ejaculation, while it was positively correlated with abnormal morphology. Body temperature was negatively correlated with BCF. Positive correlations were noticed between VAP (average path velocity) and total time of ejaculation with body acceleration features, as well as between the overall dynamic body acceleration (ODBA) and total time of ejaculation. In conclusion, the use of biomedical sensors can support the evaluation of boar sperm production capacity, providing valuable information about semen quality.

In vivo and in vitro evolution of the effects of cypermethrin on turbot (Scophthalmus maximus, Linnaeus, 1758) spermatozoa

Synthetic pyrethroid pesticide is commonly used in agricultural activities in the Black Sea region during reproduction period of turbot. In this sense, in vivo and in vitro studies have shown that cypermethrin (CYP) could be one of the environmental factors affecting decreasing turbot stocks. In this study, effects of in vivo and in vitro administration of CYP, a synthetic pyrethroid, on sperm kinematics motility (MOT), progressive motility (PM), curvilinear velocity (VCL), straight line velocity (VSL), average path velocity (VAP), linearity (LIN), straight line velocity (STR), amplitude of lateral head (ALH), beat cross frequency (BCF), oxidative stress biomarkers malondialdehyde (MDA), reduced glutathione (GSH), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and lipid peroxidation (LPO) and also histopathological alterations in gonads were investigated in spermatazoa of turbot (Schopthalmus maximus). Broodstock was supplied from culture origin and used in spawning season, additionally, two (0, 0.187 and 0.218 ppb) and three (0, 1.025, 2.05 and 4.1 ppb) different CYP concentrations were performed for in vivo and in vitro studies, respectively. In vivo and in vitro studies, significant reductions were found in sperm MOT, PM, VCL, VSL, VAP, LIN, and ALH properties depend on the increase in CYP concentrations (p < 0.05). Besides, activities of GSH, GPx, SOD, and CAT increased. In terms of histological alterations, no difference was observed among groups (0, 0.187 and 0.218 ppb) in the maturity stage of the germ cells. According to obtained results, sperm kinematics was affected significantly with increased the dose levels of CYP (p < 0.05).

Derangement in Sperm Kinematics following Pre-pubertal Exposure to Cannabis sativa: Ameliorating effect of Vitamin C.

Background of study: Motility is an important function of the male gamete which enables it to actively reach and penetrate the female gamete for fertilization. Long term use of Cannabis sativa has been associated with decreased male fertility indices including sperm motility. The characterization of the spermatozoa movement has not been evaluated especially in rats pre-pubertally exposed to cannabis sativa. Methodology: Forty pre-pubertal male rats were randomly assigned into 4 groups of ten rats each. Group 1 served as the control, group 2 was the low dose cannabis group exposed to cannabis smoke for 5 minutes daily, group 3 was the high dose cannabis group exposed to cannabis smoke for 10 minutes daily while group 4 was exposed to cannabis smoke for 10 minutes daily and treated with 2.8mg/kg vitamin C daily. Duration of administration and treatment was 28 days after which animals were euthanized and cauda epididymis from each testis was dissected out for evaluation of sperm motility indices. Resuts: The results showed significant derangement in sperm kinetics including total motility (P&lt;0.05), progressivity (P&lt;0.05), velocity of active path (P&lt;0.05), curvilinear velocity (P&lt;0.05), amplitude of lateral head (P&lt;0.05), beat cilia frequency (P&lt;0.05) and wobbling rate (P&lt;0.05) in the high dose group compared with control. These abnormal sperm kinematics were significantly ameliorated by vitamin C. Conclusion: In conclusion, cannabis smoke impairs several aspects of the kinematics of sperms in pre-pubertally exposed rats but which were ameliorated by vitamin C.

Seminal attributes of Vechur bulls (Bos indicus), the smallest humped dairy cattle

The study was undertaken at Mattupatti semen station to objectively evaluate the seminal attributes of frozenthawed semen of Vechur bulls, the smallest humped dairy cattle in the world, using the computer assisted semen analyzer (CASA) and Flow cytometer. The mean motility and progressive motility of the frozen-thawed semenestimated via CASA was 77.25±9.56 and 64.0±4.21% respectively. Mean values obtained for the sperm kinetic parameters like path velocity (VAP), progressive velocity (VSL), track speed (VCL), lateral head amplitude (ALH),beat cross frequency (BCF), straightness (STR) and linearity (LIN) were 82.56±12.12, 76.22±10.38, 107.77±21.42, 4.39±0.91, 20.87±3.81, 92.37±3.25 and 72.87±6.64% respectively. Flow cytometry analysis of the functional sperm parameters like sperm viability, live sperms with intact acrosome, sperms with normal mitochondrial membranepotential (MMP) and sperms with altered DNA were 81.67±4.55, 76.16±9.76, 82.40±5.61 and 11.75±0.3% respectively. The mean value of the HOS reactive sperms was 71.81±5.71%. Mean values for the length and width of the sperm head, length of mid-piece, tail and overall sperm length measured by computer image analysis was 8.99±0.36, 5.07±0.23, 14.21±0.35, 45.12±1.53 and 68.32±1.41 μm respectively. Study concluded that seminal attributes of Vechur bulls could be estimated with CASA and Flow cytometer and the parameters were comparable to other indigenous, crossbreds and exotic dairy cattle.

Functional Impairment of Bovine Spermatozoa by Fungicide Mancozeb: an in vitro Exposure Study

Agrochemicals have been identified as endocrine disruptors affecting many bodily functions of animals, especially the fertility of males. Evidence of various agrochemicals affecting male reproductive functions are plenty. However, evidence on disruptive effects of fungicides on sperm functions is scarce. Therefore, an in vitro study was conducted to investigate acute effects of the fungicide Mancozeb on spermatozoa functions using a bovine spermatozoa model. Semen samples were obtained from fertility proven Jersey bulls and were exposed to series of Mancozeb concentrations (0.01 μg/ml to 10 μg/ml), and the changes occurred in sperm motility parameters, acrosine proteolytic activity and acrosome integrity were assessed at different time intervals. Pure and commercial forms of Mancozeb were in the study, and sperm motility parameters were measured using computer assisted sperm analysis (CASA). Mancozeb concentration of 1 μg/ml with 2 hours exposure significantly (p≤0.05) reduced the total sperm motility and the sperm progressive motility. Velocity parameters like average path, straight line, curvy linear and amplitude lateral head displacement were significantly (p≤0.05) reduced after 2 hours in both pure and commercial Mancozeb treatments of 1 μg/ml. However, the beat cross frequency and elongation of the sperm were significantly (p≤0.05) reduced only with commercial Mancozeb. Further, the acrosome integrity and proteolytic activity of the acrosome enzymes were significantly (p≤0.05) compromised by both Mancozeb treatments. The study revealed that Mancozeb (&gt;1 ug/ml) could reduce the sperm motility and impairment of acrosome activity significantly. Nevertheless, commercial Mancozeb formulation has the highest impact on the sperm functions. Thus, Mancozeb may affect the fertilization capacity of the spermatozoa under acute exposure.

Improvement of pregnancy rate after deep uterine artificial insemination with frozen-thawed cauda epididymal spermatozoa in Hanwoo cattle

In the present study, we examined if deep uterine artificial insemination (DUAI) can improve the pregnancy rate of artificial insemination (AI) using epididymal spermatozoa (ES) in Hanwoo cattle. The estrus cycles of 88 Hanwoo cows were synchronized, and 17 cows were artificially inseminated using the DUAI method with ES, 20 cows were artificially inseminated via the uterine body (BUAI) method with ES, and as a control, 51 cows were inseminated by using the BUAI method with ejaculated spermatozoa from 1 proven bull after frozen thawing. The pregnancy rate of the DUAI method (58.8%) was higher than that of the BUAI method (25.0%, p = 0.0498). The motility of ES was examined immediately after thawing and after 3 and 6 h of incubation. The rapid progressive sperm motility of the control group was significantly higher than that of the ES group immediately after thawing and after 3 and 6 h of incubation (p < 0.05). The straight line velocity and average path velocity of the ES group after 6 h of incubation were significantly lower than those in the control group (p < 0.05). The linearity and amplitude of lateral head of ES were lower than those at 6 h (p < 0.05). The flagellar beat cross frequency and hyperactivation of ES were lower than the control spermatozoa immediately after thawing and at 3 h (p < 0.05). These motility parameters suggested that ES had a low motility and fertilization ability compared to the control spermatozoa. After frozen-thawing and 3 h of incubation, the percentage of live spermatozoa with intact acrosomes in the ES was significantly lower than that in ejaculated spermatozoa (p < 0.05). Our findings suggested that the DUAI method can overcome the low pregnancy rate of ES, despite the low motility, viability, and fertilization ability of ES.