Fertilization rate, motility, lipid peroxidation and pH changes after chilled storage of rainbow trout (Oncorhynchus mykiss) eggs and spermatozoa by a RMPI medium

The aim of this study was to evaluate the effects of Roswell Park Memorial Institute(RPMI) 1,640 medium on chilled storage of eggs and spermatozoa of rainbow trout (Oncorhynchus mykiss). After 3 days of storage, eggs in RPMI 1,640 media (pH 8.2, 9 and 10), Cortland medium and coelomic fluid were inseminated with fresh sperma‐tozoa (Experiment 1). Eggs in RPMI 1,640 medium at pH 8.2 shown the lowest thio‐barbituric acid‐reactive substances (TBARS, 0.053 ± 0.003 nmol/ml) and pH changes (from 8.20 ± 0.01 to 8.18 ± 0.01), the highest fertilization rate (82 ± 3%). Undiluted and diluted spermatozoa at ratios of 1:2 and 1:9 with RPMI 1,640 media (pH 8.2, 9 and 10) and Cortland medium were inseminated with fresh eggs (Experiment 2). Spermatozoa in RPMI 1,640 medium at pH 9 (1:9) caused the lowest TBARS content (0.037 ± 0.002 nmol/ml) and pH changes (from 9.00 ± 0.01 to 8.98 ± 0.01), the high‐est fertilization rate (77 ± 2%) and motility parameters. Based on Experiments 1 and 2, eggs and spermatozoa were stored for another 3 days in RPMI 1,640 medium at pH 8.2 and 9 (1:9) respectively (Experiment 3). Fertilization rate of storage eggs and spermatozoa in Experiment 3 was 79 ± 5%, showing successfully storage of rainbowtrout gametes with the same medium...

Effects of dietary Ferula elaeochytris root powder concentrations on haematology, serum biochemical parameters, spermatozoa parameters, and oxidative status in tissues of males goldfish (Carassius auratus)

This study evaluates the effects of Ferula elaeochytris, a traditional medicinal herb, root powder (FRP) concentrations (0, 0.5%, 1%, 0.5% and 1%) on serum biochemical parameters, hematological profiles, oxidative stress conditions in liver, muscle, gonad, testicular and stripped spermatozoa, and also spermatozoa parameters of adult male goldfish (Carassius auratus). To the best of our knowledge, this is the first time that the effects of FRP on these parameters as being associated with its essential oil contents have been determined. Some important bioactive compounds such as such as 14-beta-H-Pregna, alpha-curcumene, limonene have been determined in FRP. Following a 60-day feeding trial, blood samples were taken and then females were introduced to tanks together with changing photoperiod and water temperature for 10 days to promote spermiation. Results showed that RBC, haemoglobin, and haematocrit levels increased in 1% of FRP fed fish compared control group (P 0.05). Also serum total protein and albumin levels in this group slightly increased (P 0.05). However, blood parameters were negatively affected in 1% of FRP. Spermatozoa parameters dramatically decreased, even spermiation success could not be achieved in some fish in fish fed with 1% of FRP. Also, 1% of FRP in diet induced oxidative stress conditions in the tissues of this group. These conditions in gonad had a different pattern than those in liver in muscle. The results revealed that dietary 1% of FRP could be an advantageous additive while FRP at levels of >0.5% in diet was useless for or has deleterious effects on fish health and reproduction. Based on these results, we conclude that a supplement suitable concentration and analysis of major compounds of medicinal herbs which would use should be taken into consideration when planning to use them in fish diets

Comparison of milt and blood parameters including testosterone and gonadotropins levels of sex-reversed female rainbow trout (Oncorhynchus mykiss) by different steroid hormones

All-female progeny is preferable in the reproduction of rainbow trout. Using sex-reversed (SR) female rainbow trout by steroid hormones as broodstock in the reproduction is the most appropriate way to all-female progeny. This study compared mature SR female rainbow trout by ethisterone (ET), 11 beta-Hydroxyandrostenedione (HD) and 17 alpha-Methyltestosterone (MT) in terms of spermatological parameters and health status, for the first time. Spermatozoa concentration, motility characteristics, seminal plasma pH, testosterone and gonadotropins in seminal plasma and blood serum, blood parameters of SR females were evaluated, comparing with testicular and stripped milt, and blood from normal males (NM). The SR females by HD had greater gonadosomatic index and better motility characteristics compared with other groups. Also, blood parameters of this group were more similar to those in NM. The SR females by ET had the lowest haematocrit and the highest glucose and activities of the enzymes compared with the other groups. The measured hormones were significantly the lowest in both seminal plasma and serum of the ET group. These findings suggested that using of HD for obtaining SR female should be preferred due to having the better quality spermatozoa and health status

DNA damage, oxidative stress, decreased viability and motility in common carp (Cyprinuscarpio l.) spermatozoa induced by tryptophan, phenylalanine and cysteine amino acids during short-term storage

Free amino acids are found in spermatozoa and seminal plasma. Their qualitative and quantitative presence have been affected by many factors. Under in vitro storage conditions, their effects on spermatozoa in animal species and also human have been concerned over the last decades. In this study, the effects of tryptophan, phenylalanine and cysteine on DNA integrity, lipid peroxidation, viability and motility parameters in spermatozoa of the common carp (Cyprinuscarpio) were assessed during short-term storage. Spermatozoa were incubated at 1, 5, 25, and 50 mM of these amino acids in vitro at 4 °C in a time‐dependent manner (6, 24, and 48 h). The supplementation of cysteine, phenylalanine and >5 mM of tryptophan to the incubation medium attenuated the spermatozoa motility parameters and viability. Particularly, >1 mM of cysteine completely inhibited progressive motility and decreased viability to 5 mM of tryptophan to the incubation medium attenuated the spermatozoa motility parameters and viability. Particularly, >1 mM of cysteine completely inhibited progressive motility and decreased viability to <50%. Cysteine and phenylalanine at all concentrations significantly caused the increases in lipid peroxidation products in spermatozoa and seminal plasma, and hereby induced the percentage of DNA damage in spermatozoa. Cysteine had more detrimental effects than phenylalanine and tryptophan. Our data provide evidence that these amino acids are not beneficial for the maintaining of carp spermatozoa at 4 °C

Effect of alpha-lipoic acid on oxidative stress, viability and motility in the common carp (Cyprinus carpio) spermatozoa after short-term storage and cryopreservation

İnanan, Burak EvrenAs known for different metabolic functions, α-lipoic acid (ALA) has been tested for spermatozoa preservation of animals as well as of human, but not for fish spermatozoa. The present study determined the effects of ALA on short and long-term (cryopreservation) preservation of common carp (Cyprinus carpio) spermatozoa, for the first time. For that, spermatozoa were diluted in extenders containing 0 (control), 0.025, 0.05, 0.1, 0.5, 1, 2, 5, and 10 mM of ALA concentrations in both short-term preservation and cryopreservation. Spermatozoa motility parameters by computer-assisted semen analysis, viability, lipid peroxidation and catalase activity in spermatozoa were conducted in both 2nd and 120th hours of short-term storage and post-thaw samples. Higher percentages of total spermatozoa motility (80 ± 3) and viability (87 ± 3) were observed in 0.5 mM ALA group after 120 h of incubation. In post-thaw samples, higher percentages of these parameters were in 1 mM ALA group (74 ± 3 and 83 ± 2, respectively). Moreover, the results have shown that the addition of ALA until concentrations of 2 mM improved especially spermatozoa curvilinear velocity, maintained viability, and suppressed excessive lipid peroxidation during the preservations. In conclusion, the additions of 0.5 mM ALA for short-term preservation and 1 mM ALA for cryopreservation were the optimal concentrations, and shown the protective effects on common carp spermatozoa, when considering all measured parameters together

In vitro effects of fenugreek, sunflower, green cardamom and seed extracts on motility parameters and oxidative stress of common carp (Cyprinus carpio L.) spermatozoa

İnanan, Burak Evren( Aksaray, Yazar )Bu çalışmanın amacı, üç farklı bitki familyasına ait olan kakule (Trigonella foenum-graecum, Fabaceae), Ayçiçeği (Helianthus annuus, Asteraceae) ve çemen otu (Elettaria cardamomum, Zingiberaceae) tohumu ekstraktlarının in vitro koşullar altında, sazan balığı (Cyprinus carpio L.) sperm hücrelerinin motilite parametrelerine ve oksidatif stres durumlarına etkilerini belirlemektir. Bu amaçla, %0,1, %0,5, %1 ve %2 oranlarındaki tohum ekstratları ile muamele edilen sperm örnekleri 4°C’de muhafaza edilerek, 2. ve 48. saatte ölçümleri yapılmıştır. Sperm motilite parametrelerinden kavisli hareket hızı (VCL), ortalama hızı (VAP), ileri doğru doğrusal hareket hızı (VSL), doğruluk (STR), doğrusallık (LIN), sperm başı titreşim hareketi (WOB) tayinleri bilgisayar yardımlı sperm analizi (CASA) sistemi ile gerçekleştirilmiştir. Elde edilen sonuçlara göre, Ayçiçeği ve çemen otu tohum ekstraktları, test edilen oranların tamamında motilite parametrelerini azaltıcı yönde tesir etmiştir. Buna karşın, 48. saatte %0,5 kakule tohumu ekstraktı kontrol grubu dâhil tüm gruplara kıyasla motilite parametrelerini yükseltmiştir. Ayrıca bu grupta, oksidatif stres göstergesi olarak ölçülen TBARS değerleri azalmıştır. %2 tohum ekstraktlarının tümü negatif olarak sperm örneklerini etkilemiştir. Özellikle, %2 Ayçiçeği tohum ekstratı grubu yüksek oksidatif strese neden olmuştur. Bu çalışmanın sonuçları, 4°C’de sazan balığı sperm muhafazasında, ayçiçeği ve çemen otuna göre kakule tohum ekstraktının kullanımının daha uygun olduğunu göstermiştir.The aim of the present study is to assess the effects Fenugreek (Trigonella foenum-graecum, Fabaceae), sunflower (Helianthus annuus, Asteraceae), green cardamom (Elettaria cardamomum, Zingiberaceae) and seed extracts, which they belonged to three different plant families, on common carp (Cyprinus carpio L.) spermatozoa motility parameters and oxidative stress conditions. For this reason, sperm samples treated with seed extracts at the ratios of 0.1%, %0.5, 1%, and 2%, were incubated at 4°C and sampled at 2nd and 48th hours. Among sperm motility parameters, curvilinear velocity (VCL), average path velocity (VAP), straight-line velocity (VSL), straightness STR), linearity (LIN), and wobble (WOB) were determined by computer-assisted sperm analysis (CASA). According to the results, sunflower and green cardamom seed extracts have attenuated motility parameters at all tested concentrations. However, motility parameters measured in %0.5 of fenugreek seed extract were higher than all other groups including the control at 48th hour. Also, TBARS values as an oxidative stress indicator in this group were decreased. 2% of all seed extracts had negative effects on the sperm samples. Particularly, 2% sunflower seed extract caused the higher oxidative stress. These results indicate that fenugreek seed extract is more proper for the maintenance of common carp spermatozoa at 4°C, comparing to those of sunflower and green cardamom

Seasonal and age-related changes in semen characteristics and composition of seminal plasma in sex-reverse female rainbow trout (Oncorhynchus mykiss) in comparison with normal males

WOS: 000447110900019PubMed: 30077522All-male or all-female progeny is preferable in reproduction of fish species. In rainbow trout, all female progeny is desirable, and the most common and healthful approach is to use sex reversed (SR) females as broodstock by including androgen hormones through the diet at the larval stage. The purpose of this study was to determine the differences and changes of seminal plasma ions (Na+ K+ Ca2+, Mg2+ and Cl-) and pH in 2 to 3 and 3 to 4 year-old SR females, compared with 2 to 3 year-old normal (N) males at the beginning, middle, and end of the spawning season (SS). Additionally, osmolality, spermatozoa concentration, and sperm motility characteristics were determined. Percentages of sperm motility of SR females were negatively correlated with seminal plasma pH and sperm concentrations (r = -0.479 and r = -0.534, respectively, P 0.05). Furthermore, using of 2 to 3 year-old rather than 3 to 4 year-old SR females for reproduction could provide an advantage in terms of sperm motility characteristics. These findings could be useful for not only understanding of SR of semen from sexually reversed females but also for design of a maturation medium both for further studies and aquaculture production.Mugla Sitki Kocman University Research Fund [2012/92]; Scientific and Technical Research Council of Turkey (TUBITAK) [1130606]This research was supported by Mugla Sitki Kocman University Research Fund (Grant number: 2012/92) and partially by the Scientific and Technical Research Council of Turkey (TUBITAK) (Grant number: 1130606). We would like to thank all the personnel in the fish farms who assisted in the sampling

Effect of semen extender supplementation with cysteine on postthaw sperm quality DNA damage and fertilizing ability in the common carp Cyprinus carpio

Amino acids have an important biological role for prevention of cell damage during cryopreservation. The objective of this study is to determine the effects of cysteine on postthaw sperm motility, duration of sperm motility, DNA damage, and fertility in the common carp (Cyprinus carpio). Sperm collected from 10 individuals was cryopreserved in extenders containing different cysteine concentrations (2.5, 5, 10, and 20 mM). Semen samples diluted at the ratio of 1:9 by the extenders were subjected to cryopreservation. After dilution, the semen was aspirated into 0.25-mL straws; the straws were placed on the tray, frozen in nitrogen vapor, and plunged into liquid nitrogen. DNA damage was evaluated by comet assay after cryopreservation. Our results indicated that an increase in the concentration of cysteine caused a significant increase in the motility rate and duration of sperm in the common carp (C carpio; P < 0.05). Comparing all concentrations of cysteine, the best concentration of cysteine was 20 mM. Higher postthaw motility (76.00 ± 1.00%) and fertilization (97.00 ± 1.73%) rates were obtained with the extender at the concentration of 20 mM. Supplementation of the extender with cysteine was increased the fertilization and hatching rate and decreased DNA damage. Consequently, cysteine affected the motility, fertilization, and DNA damage positively, and extenders could be supplemented with cysteine

Effects of semen extender supplemented with L methionine and packaging methods straws and pellets on post thaw goldfish Carassius auratus sperm quality and DNA damage

BACKGROUND: Amino acids protect spermatozoa against cell damage during cryopreservation due to have antioxidant property and found in seminal plasma at high concentration. OBJECTIVE: The aim of the present work was to analyse the effect of extender supplementation with L-methionine on post-thawed sperm motility, duration and DNA damage and also it was tested the feasibility of using straws and pellets method for the cryopreservation of goldfish (Carassius auratus) sperm. MATERIALS AND METHODS: Extenders were supplemented with different L-methionine concentrations of 1 mM; 1.5 mM; 3 mM; 6 mM. Semen samples diluted at the ratio of 1 to 9 by the extenders were subjected to cryopreservation. After dilution the semen was aspirated into 0.25 ml straws and 0.1 ml pellets, the straws and pellets were placed on the tray, frozen in nitrogen vapor and plunged into liquid nitrogen. DNA damage was determined with comet assay after cryopreservation. RESULTS: Our results indicated that an increase in the concentration of L-methionine caused a significant increase in the motility rate and duration of sperm in goldfish (C. auratus) (p<0.05). In addition, duration and percentage of motility in pellets were higher than in straws. Comparing all concentrations of L-methionine, the best concentration of L-methionine was 1.5 mM. Highest post-thaw motility (45.00 +/- 7.07%) and duration of motility (17.00 +/- 0.71s) were obtained with the extender at concentration 1.5 mM in pellets. Addition of the extender with L-methionine was reduced DNA damage compared to control group. CONCLUSION: Consequently, pellets could be used for goldfish sperm cryopreservation and the tested amino acid affected the motility parameters, and semen extenders could be supplement with L-methionine