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Sample records for abalone sperm lysin

  1. Lead behavior in abalone shell

    Science.gov (United States)

    Hirao, Yoshimitsu; Matsumoto, Akikazu; Yamakawa, Hiroshi; Maeda, Masaru; Kimura, Kan

    1994-08-01

    In order to gain information about the behavior of heavy metals in biological assimilation processes in a marine food chain and to investigate the possibility that lead pollution in a marine environment can be estimated by measurement of a small number of key materials from such a food chain, muscle and shell were analyzed from abalone ( Haliotis) from a shallow water locality in a Japanese coastal region. Lead concentrations in muscle were about 26 ppb for abalone of approximately 3 years old and decreased systematically with increasing age of animals sampled, to about 3.3 ppb for a specimen approximately 8 years old. Lead concentrations in shell material gradually decreased also, from 150 ppb to 82 ppb in the oldest specimen. The decrease of concentration in tissues with increasing age indicates that a mechanism for exclusion of lead during tissue growth becomes more efficient with age. Along the food chain in which abalone is the final stage, lead was enriched at the first stage, from seawater to algae, by a factor of 100. Lead was diminished at all subsequent stages of the chain. Tissue of artificially cultured abalone had four times higher lead values compared to abalone grown in natural conditions, and this appears to reflect the fact that lead concentration was three times higher in seawater in the cultured environment.

  2. Evidence of green fluorescent protein and growth hormone expression in red abalone (Haliotis rufescens larvae

    Directory of Open Access Journals (Sweden)

    Mancilla-Sánchez Edgar

    2017-02-01

    Full Text Available The red abalone Haliotis rufescens is a highly appreciated mollusk in the national and international markets. Due to its natural over-exploitation and low growth rate, several genetic improvements were made, however special efforts are needed to increase its production. This study presents transgenic abalone’s larvae expressing green fluorescent protein (GFP fused to Cobia (Rachycentron canadum Growth Hormone (GH using sperm media transgenesis technique (SMT, pAcGFP1-N vector under the control of cytomegalovirus (CMV promoter. Sperms were exposed to three voltages (0.5, 0.75 and 1.0 Kv using a micropulser electroporator (Bio-Rad®. The highest GFP-GH expression average (40% was obtained in abalone larvae at 0.75 v. GFP and GH transgenes were positively detected by PCR, western blot and confocal microscope, respectively.

  3. Overexploitation of Abalone at Libong Island, Trang Province, Southern Thailand

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    Chanyut Sudtongkong

    2012-09-01

    Full Text Available Abalone is generally known as a fishery resource of high economic value. The wild abalone from Libong Island iswidely known for its potential as a cocktail-size and high-quality broodstock for hatcheries. The high market price andexternal demand have encouraged local fisherman to catch the wild abalone without proper management, resulting in a nearextinction crisis in the abalone population in this area. The present evaluation of abalone management at Libong Island,Trang Province, Thailand, was conducted using local user perceptions. Sixteen performance indicators included effectiveness indicators, equity indicators, and sustainable indicators. These were measured to determine whether the abalonemanagement activities had achieved the set objectives in terms of better conditions for abalone cultivation and sustainability.The results revealed that the abalone population has undergone degrading and decline due to lack of proper managementmeasures in this area. The findings suggest that practical management is needed for the abalone population at Libong Island.

  4. Abalone farm discharges the withering syndrome pathogen into the wild

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    Kevin eLafferty

    2013-12-01

    Full Text Available An intracellular bacterium Candidatus Xenohaliotis californiensis, also called Withering-Syndrome Rickettsia-Like Organism (WS-RLO, is the cause of mass mortalities that are the chief reason for endangerment of black abalone (Haliotis cracherodii. Using a real-time PCR assay, we found that a shore-based abalone farm in Santa Barbara, California, discharged WS-RLO DNA into the ocean. Several other shore-based abalone farms discharge effluent into critical habitat for black abalone in California and this might affect the recovery of wild black abalone. Existing regulatory frameworks exist that could help protect wild species from pathogens released from shore-based aquaculture.

  5. First attempts to cryopreserve red abalone (Haliotis rufescens oocytes

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    Ramírez Torrez, A.

    2015-01-01

    Full Text Available Overall, few advances in the cryopreservation of complex cells such as oocytes, embryo or tissue have been registered and in less quantity have been reported for aquatic species. Abalone has high economic interest worldwide and the conservation of abalone germplasm may help to enhance its culture and develop repopulation programs. In this work, we reported the cytotoxic effect of two concentration of trehalose (0.2 and 0.4 M on red abalone oocytes incubated for 10, 15 and 20 min. Also, we reported the cryopreservation of red abalone oocytes using a 3-steps cryopreservation protocol and 5 thawing protocols. Significant differences on cytotoxic effect were found (p<0.01. However, none of the cryoprotectant was optimum to cryopreserve red abalone oocyte. In conclusion, it is necessary to find an appropriate method to dehydrate or make the cryoprotectant penetrate on the abalone oocyte before proceeding to cryopreservation.

  6. Cryopreservation of microencapsulated canine sperm.

    Science.gov (United States)

    Shah, Shambhu; Otsuki, Tsubasa; Fujimura, Chika; Yamamoto, Naoki; Yamashita, Yasuhisa; Higaki, Shogo; Hishinuma, Mitsugu

    2011-03-01

    The objective was to develop a method for cryopreserving microencapsulated canine sperm. Pooled ejaculates from three beagle dogs were extended in egg yolk tris extender and encapsulated using alginate and poly-L-lysine at room temperature. The microcapsules were cooled at 4 °C, immersed in pre-cooled extender (equivalent in volume to the microcapsules) to reach final concentration of 7% (v/v) glycerol and 0.75% (v/v) Equex STM paste, and equilibrated for 5, 30 and 60 min at 4 °C. Thereafter, microcapsules were loaded into 0.5 mL plastic straws and frozen in liquid nitrogen. In Experiment 1, characteristics of microencapsulated canine sperm were evaluated after glycerol addition at 4 °C. Glycerol exposure for 5, 30 and 60 min did not significantly affect progressive motility, viability, or acrosomal integrity of microencapsulated sperm compared with pre-cooled unencapsulated sperm (control). In Experiment 2, characteristics of frozen-thawed canine microencapsulated sperm were evaluated at 0, 3, 6, and 9 h of culture at 38.5 °C. Pre-freeze glycerol exposure for 5, 30, and 60 min at 4 °C did not influence post-thaw quality in unencapsulated sperm. Post-thaw motility and acrosomal integrity of microencapsulated sperm decreased more than those of unencapsulated sperm (P < 0.05) following glycerol exposure for 5 min. However, motility, viability and acrosomal integrity of microencapsulated sperm after 30 and 60 min glycerol exposure were higher than unencapsulated sperm cultured for 6 or 9 h (P < 0.05). In conclusion, since microencapsulated canine sperm were successfully cryopreserved, this could be a viable alternative to convention sperm cryopreservation in this species. Copyright © 2011 Elsevier Inc. All rights reserved.

  7. Heat inactivation of a norovirus surrogate in cell culture lysate, abalone meat, and abalone viscera.

    Science.gov (United States)

    Park, Shin Young; Bae, San-Cheong; Ha, Sang-Do

    2015-03-01

    The current study examined the effects of temperature and heat treatment duration on murine norovirus-1 (MNV-1) from both viral cell culture lysate (7-8 log10 PFU) and experimentally contaminated abalone meat and viscera (5-6 log10 PFU) as a model of human norovirus (NoV). MNV-1 titers in cell culture lysate, abalone meat, and abalone viscera were gradually reduced to 1.93-4.55, 1.79-3.00, and 2.26-3.26 log10 PFU/ml, respectively, after treatment at 70 °C for 1-10 min. Treatment at 85 °C for 1-5 min gradually reduced MNV-1 titers in abalone meat to 2.71-4.15 log10 PFU/ml. MNV-1 titers in abalone viscera were gradually reduced to 2.91-3.46 log10 PFU/ml after treatment at 85 °C for 1-3 min. No significant difference (P > 0.05) was found in MNV-1 titers in the abalone meat and viscera among treatment groups (70 °C for 5 min, 70 °C for 3 min, and 85 °C for 1 min). Complete inactivation of MNV-1 in cell culture lysate was determined at 85 °C for ≥1 min and 100 °C for ≥0.5 min. Complete inactivation of MNV-1 in abalone was determined at 100 °C for ≥0.5 min for meat, and 85 °C for 5 min and 100 °C for ≥0.5 min for viscera. At treatments at 70 °C, the Td-values (3 log reduction time) were significantly lower (P 0.05) between abalone meat (1.78) and abalone viscera (1.33) at treatments at 85 °C. This study suggests that 100 °C for ≥0.5 min could potentially be used to inactivate NoV in molluscan shellfishes, including viscera.

  8. Short Communication Towards an abalone weaning diet: evaluation ...

    African Journals Online (AJOL)

    Survival of post-settlement abalone when 'weaning' them from diatoms onto macroalgae/artificial feed remains unpredictable for farmers. It is hypothesised that spirulina algae, which are high in protein, may be a suitable feed ingredient for weaning abalone. Over a period of 27 days, the growth and survival of juvenile (4–6 ...

  9. Spionid polychaetes infesting cultured abalone Haliotis midae in ...

    African Journals Online (AJOL)

    CA Simon, A Ludford, S Wynne. Abstract. A study was conducted to identify and quantify the spionid polychaetes that infest cultured abalone Haliotis midae at aquaculture facilities on the west (Farm A), south (Farms B and C) and ... The abalone were infested by Dipolydora capensis, Polydora hoplura and a Boccardia sp.

  10. Short Communication Growth of post-weaning abalone Haliotis ...

    African Journals Online (AJOL)

    The effect of five formulated feeds supplemented with fresh wild seaweed on the growth of post-weaning juvenile abalone Haliotis midae (6–20 mm shell length) was investigated by means of a growth trial at a commercial abalone farm over a period of 11 months. The experiment included 10 diet treatments with two ...

  11. Feeding preferences and the nutritional value of tropical algae for the abalone Haliotis asinina.

    Directory of Open Access Journals (Sweden)

    Alex R Angell

    Full Text Available Understanding the feeding preferences of abalone (high-value marine herbivores is integral to new species development in aquaculture because of the expected link between preference and performance. Performance relates directly to the nutritional value of algae--or any feedstock--which in turn is driven by the amino acid content and profile, and specifically the content of the limiting essential amino acids. However, the relationship between feeding preferences, consumption and amino acid content of algae have rarely been simultaneously investigated for abalone, and never for the emerging target species Haliotis asinina. Here we found that the tropical H. asinina had strong and consistent preferences for the red alga Hypnea pannosa and the green alga Ulva flexuosa, but no overarching relationship between protein content (sum of amino acids and preference existed. For example, preferred Hypnea and Ulva had distinctly different protein contents (12.64 vs. 2.99 g 100 g(-1 and the protein-rich Asparagopsis taxiformis (>15 g 100 g(-1 of dry weight was one of the least preferred algae. The limiting amino acid in all algae was methionine, followed by histidine or lysine. Furthermore we demonstrated that preferences can largely be removed using carrageenan as a binder for dried alga, most likely acting as a feeding attractant or stimulant. The apparent decoupling between feeding preference and algal nutritive values may be due to a trade off between nutritive values and grazing deterrence associated with physical and chemical properties.

  12. Mortality of the abalone Haliotis mariae (Haliotidae: Mollusca in aquaculture

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    Gilha Yoon

    2015-01-01

    Full Text Available The Omani abalone, Haliotis mariae, is the only species of abalone found in Omani waters. Given the rarity of this species and the high price it can fetch on the market (US$ 150 kg-1 dry weight, the wild abalone fishery in the Dhofar region has been regarded as a valuable income source for the past decade. The present study was undertaken set to investigate the mortality of abalone held at the Mirbat Research Center, through bacteriological and histopathological based investigations and challenge tests. Only the adult wild abalone that had been kept for a year in the hatchery, visually, appeared to be clear of disease symptoms. Infected individuals typically were swollen around the mouth, had fluid tinged with blood, bubbles in the intestines, and, very weak adhesive strength. The foot area (muscle of diseased animals was noticeably very soft and individuals that were seen lying upside down on the bottom of the tank subsequently died. On dissection, the intestinal organs released bubbles and a foul smelling odour. Identification of the isolated bacteria using various identification methods indicated that individuals were infected with Staphylococcus sciuri. Histopathology of infected individuals revealed spongiosis of the tissues with evident bacterial infection. Neither of these histopathological conditions were seen in healthy abalone. The study concludes that the bacterium Staphylococcus sciuri may be the likely cause of abalone mortalities.

  13. Analysis of microbiota on abalone (Haliotis discus hannai) in South Korea for improved product management.

    Science.gov (United States)

    Lee, Min-Jung; Lee, Jin-Jae; Chung, Han Young; Choi, Sang Ho; Kim, Bong-Soo

    2016-10-03

    Abalone is a popular seafood in South Korea; however, because it contains various microorganisms, its ingestion can cause food poisoning. Therefore, analysis of the microbiota on abalone can improve understanding of outbreaks and causes of food poisoning and help to better manage seafood products. In this study, we collected a total of 40 abalones from four different regions in March and July, which are known as the maximum abalone production areas in Korea. The microbiota were analyzed using high-throughput sequencing, and bacterial loads on abalone were quantified by real-time PCR. Over 2700 species were detected in the samples, and Alpha- and Gammaproteobacteria were the predominant classes. The differences in microbiota among regions and at each sampling time were also investigated. Although Psychrobacter was the dominant genus detected on abalone in both March and July, the species compositions were different between the two sampling times. Five potential pathogens (Lactococcus garvieae, Yersinia kristensenii, Staphylococcus saprophyticus, Staphylococcus warneri, and Staphylococcus epidermidis) were detected among the abalone microbiota. In addition, we analyzed the influence of Vibrio parahaemolyticus infection on shifts in abalone microbiota during storage at different temperatures. Although the proportion of Vibrio increased over time in infected and non-infected abalone, the shifts of microbiota were more dynamic in infected abalone. These results can be used to better understand the potential of food poisoning caused by abalone consumption and manage abalone products according to the microbiota composition. Copyright © 2016 Elsevier B.V. All rights reserved.

  14. Gradual Ordering in Red Abalone Nacre

    Energy Technology Data Exchange (ETDEWEB)

    Gilbert, P. U. P. A.; Metzler, Rebecca A.; Zhou, Dong; Scholl, Andreas; Doran, Andrew; Young, Anthony; Kunz, Martin; Tamura, Nobumichi; Coppersmith, Susan N.

    2008-09-03

    Red abalone (Haliotis rufescens) nacre is a layered composite biomineral that contains crystalline aragonite tablets confined by organic layers. Nacre is intensely studied because its biologically controlled microarchitecture gives rise to remarkable strength and toughness, but the mechanisms leading to its formation are not well understood. Here we present synchrotron spectromicroscopy experiments revealing that stacks of aragonite tablet crystals in nacre are misoriented with respect to each other. Quantitative measurements of crystal orientation, tablet size, and tablet stacking direction show that orientational ordering occurs not abruptly but gradually over a distance of 50 {micro}m. Several lines of evidence indicate that different crystal orientations imply different tablet growth rates during nacre formation. A theoretical model based on kinetic and gradual selection of the fastest growth rates produces results in qualitative and quantitative agreement with the experimental data and therefore demonstrates that ordering in nacre is a result of crystal growth kinetics and competition either in addition or to the exclusion of templation by acidic proteins as previously assumed. As in other natural evolving kinetic systems, selection of the fastest-growing stacks of tablets occurs gradually in space and time. These results suggest that the self-ordering of the mineral phase, which may occur completely independently of biological or organic-molecule control, is fundamental in nacre formation.

  15. Sperm competition, sperm numbers and sperm quality in muroid rodents.

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    Laura Gómez Montoto

    Full Text Available Sperm competition favors increases in relative testes mass and production efficiency, and changes in sperm phenotype that result in faster swimming speeds. However, little is known about its effects on traits that contribute to determine the quality of a whole ejaculate (i.e., proportion of motile, viable, morphologically normal and acrosome intact sperm and that are key determinants of fertilization success. Two competing hypotheses lead to alternative predictions: (a sperm quantity and quality traits co-evolve under sperm competition because they play complementary roles in determining ejaculate's competitive ability, or (b energetic constraints force trade-offs between traits depending on their relevance in providing a competitive advantage. We examined relationships between sperm competition levels, sperm quantity, and traits that determine ejaculate quality, in a comparative study of 18 rodent species using phylogenetically controlled analyses. Total sperm numbers were positively correlated to proportions of normal sperm, acrosome integrity and motile sperm; the latter three were also significantly related among themselves, suggesting no trade-offs between traits. In addition, testes mass corrected for body mass (i.e., relative testes mass, showed a strong association with sperm numbers, and positive significant associations with all sperm traits that determine ejaculate quality with the exception of live sperm. An "overall sperm quality" parameter obtained by principal component analysis (which explained 85% of the variance was more strongly associated with relative testes mass than any individual quality trait. Overall sperm quality was as strongly associated with relative testes mass as sperm numbers. Thus, sperm quality traits improve under sperm competition in an integrated manner suggesting that a combination of all traits is what makes ejaculates more competitive. In evolutionary terms this implies that a complex network of genetic

  16. NUTRITION REQUIREMENT OF CULTURED ABALONE POST LARVAE AND JUVENILES: A REVIEW

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    Wa Iba

    2008-06-01

    Full Text Available Abalone aquaculture attracts wide attention nowadays because of its high market value and depleted of wild stocks. China and Japan are the main producer of abalone from aquaculture followed by other countries such as New Zealand, Australia and US. Most of cultured abalone are temperate species but considerable research efforts have been made to culture tropical abalone in the Philippines, Thailand, and Indonesia. Most of abalone culture still depends on natural food but recent studies have been conducted to develop artificial diet for abalone. This will not only reduce wild harvest of macrolagae but also develop high nutrition diet at low cost. Successful abalone aquaculture is determined by correct nutrition supplement in the diet. As other cultured animal, abalone requires balanced nutrition of carbohydrate, protein, lipid, vitamins, and minerals. As herbivores, abalone can utilize carbohydrate efficiently as energy source and thus only requires low level of protein (range from 27%—40%. Lipid requirements range from 3%—5% while some minerals such as calcium and phosphorus in artificial feed are only needed in small amount, 0.5% of calcium in diets and 0.7% of phosphorus in the diet can improve the growth rate of abalone. There is not available information of vitamin upplementation in the diet but it is suggested that natural food meets the requirement.

  17. Concentration and retention of Toxoplasma gondii surrogates from seawater by red abalone (Haliotis rufescens)

    Science.gov (United States)

    Schott, Kristen C; Krusor, Colin; Tinker, M. Tim; Moore, James G.; Conrad, Patricia A.; Shapiro, Karen

    2016-01-01

    Small marine snails and abalone have been identified as high- and low-risk prey items, respectively, for exposure of threatened southern sea otters to Toxoplasma gondii, a zoonotic parasite that can cause fatal encephalitis in animals and humans. While recent work has characterized snails as paratenic hosts for T. gondii, the ability of abalone to vector the parasite has not been evaluated. To further elucidate why abalone predation may be protective against T. gondii exposure, this study aimed to determine whether: (1) abalone are physiologically capable of acquiring T. gondii; and (2) abalone and snails differ in their ability to concentrate and retain the parasite. Abalone were exposed to T. gondii surrogate microspheres for 24 h, and fecal samples were examined for 2 weeks following exposure. Concentration of surrogates was 2–3 orders of magnitude greater in abalone feces than in the spiked seawater, and excretion of surrogates continued for 14 days post-exposure. These results indicate that, physiologically, abalone and snails can equally vector T. gondii as paratenic hosts. Reduced risk of T. gondii infection in abalone-specializing otters may therefore result from abalone's high nutritional value, which implies otters must consume fewer animals to meet their caloric needs.

  18. Sperm Production Rate, Gonadal and Extragonadal Sperm ...

    African Journals Online (AJOL)

    Five healthy West African Dwarf (WAD) rams, 1.5 to 2.5 years of age and weighing between 15 kg to 20 kg were used to determine daily sperm production, gonadal and exragonadal sperm reserves. Gonadal and extragonadal sperm reserves were estimated by the haemocytometric method, while the daily sperm production ...

  19. The steadily increasing demand for South African abalone Haliotis ...

    African Journals Online (AJOL)

    denise

    of kelp beds, with potentially large ecological and commercial effects. The aims of this investigation were to estimate the potential harvest (standing stock) ...... DAY, E. 1998 — Ecological interactions between abalone (Haliotis midae) juveniles and sea urchins (Parechinus angulosus), off the south-west coast of South Africa.

  20. Abalone Haliotis spp. are in considerable demand worldwide ...

    African Journals Online (AJOL)

    spamer

    tauropine, as a result of condensation reactions be- tween pyruvate and the amino acids arginine, glycine, alanine and tauropine respectively, catalysed ...... 20: 567–570. JAMES, D. G. and J. OLLEY 1970 — Moisture and pH changes as criteria of freshness in abalone and their relationship to texture of the canned product.

  1. Abalone Haliotis spp. are in considerable demand worldwide ...

    African Journals Online (AJOL)

    spamer

    Abalone Haliotis spp. are in considerable demand worldwide, especially in the Far East. This has en- couraged industries that involve not only harvesting natural populations but also, as such populations have declined, setting up artificial cultures of appropriate species (Lo-Chai Chen 1990, Britz 1996, Knauer et al. 1996).

  2. Assessing the ecosystem effects of the abalone Haliotis midae from ...

    African Journals Online (AJOL)

    The South African abalone Haliotis midae is commercially exploited and seriously threatened by overfishing. This not only affects the species itself but potentially the functioning of the ecosystem because of associated changes in community structure. The nature of effects that can follow the loss or reduction of a species ...

  3. Book Review: Abalone of the World: Biology, Fisheries and Culture ...

    African Journals Online (AJOL)

    Book Title: Abalone of the World: Biology, Fisheries and Culture. Book Authors: Edited by S.A. Shepherd, M.J. Tegner & S.A. Guzman del Proo. Fishing News Books (Division of Blackwell Scientific. Publications Ltd.) Oxford, U.K. (1992). 608 pages. ISBN 0-85238-181-6.

  4. Book Review Abalone of the World: Biology, Fisheries and Culture

    African Journals Online (AJOL)

    Although this book originated from the First International. Symposium on Abalone Biology, which was held in La Paz,. Mexico in 1989, it is far more than a collection of symposium papers. The symposium papers are included in the volume but, in addition to this, the editors solicited review articles on several of the more ...

  5. Book Review Abalone of the World: Biology, Fisheries and Culture

    African Journals Online (AJOL)

    Edited by S.A. Shepherd, M.J. Tegner and S.A.. Guzman del Proo. Fishing News Books (Division of Blackwell Scientific. Publications ltd.) Oxford, U.K. (1992). 608 pages. Price: £65. ISBN 0-85238-181-6. Although this book originated from the First International. Symposium on Abalone Biology, which was held in La Paz,.

  6. Aquaculture-oriented genetic researches in abalone: Current status ...

    African Journals Online (AJOL)

    Taghwo

    2013-06-26

    Jun 26, 2013 ... *CS, Cold shock; HS, heat shock; HPS, hydropressure shock; CB, cytochlasin B; 6-DMAP, 6-dimethylaminopurine. ..... Induction of gynogenetic diploid by inhibiting second meiosis in the Pacific abalone. Nippon Suisan Gakkaishi 56: 1755-1763. Fujino K, Arai K, Kudo M, Suzuki T, Masumori M, Takahashi K ...

  7. The South African commercial abalone Haliotis midae fishery began ...

    African Journals Online (AJOL)

    spamer

    The South African commercial abalone Haliotis midae fishery began in 1949 and soon covered approxi- mately 580 km of coastline from Cape Columbine to. Quoin Point (Fig. 1). The catches in the 1960s were extremely high, and certainly greater than were sus- tainable, but fell dramatically in the 1970s. Although.

  8. Abalone water-soluble matrix for self-healing biomineralization of tooth defects

    Energy Technology Data Exchange (ETDEWEB)

    Wen, Zhenliang [Institute of Biomedical and Pharmaceutical Technology, Fuzhou University, Fuzhou 350002 (China); Chen, Jingdi, E-mail: ibptcjd@fzu.edu.cn [Institute of Biomedical and Pharmaceutical Technology, Fuzhou University, Fuzhou 350002 (China); Wang, Hailiang [The Affiliated Stomatological Hospital, Fujian Medical University, Fuzhou 350002 (China); Zhong, Shengnan; Hu, Yimin; Wang, Zhili [Institute of Biomedical and Pharmaceutical Technology, Fuzhou University, Fuzhou 350002 (China); Zhang, Qiqing, E-mail: zhangqiq@126.com [Institute of Biomedical and Pharmaceutical Technology, Fuzhou University, Fuzhou 350002 (China); Institute of Biomedical Engineering, Chinese Academy of Medical Science & Peking Union Medical College, Tianjin 300192 (China)

    2016-10-01

    Enamel cannot heal by itself if damaged. Hydroxyapatite (HAP) is main component of human enamel. Formation of enamel-like materials for healing enamel defects remains a challenge. In this paper, we successfully isolated the abalone water-soluble matrix (AWSM) with 1.53 wt% the abalone water-soluble protein (AWSPro) and 2.04 wt% the abalone water-soluble polysaccharide (AWSPs) from abandoned abalone shell, and self-healing biomineralization of tooth defects was successfully achieved in vitro. Based on X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), hot field emission scanning electron microscopy (HFESEM) and energy dispersive spectrometer (EDS) analysis, the results showed that the AWSM can efficiently induce remineralization of HAP. The enamel-like HAP was successfully achieved onto etched enamel's surface due to the presence of the AWSM. Moreover, the remineralized effect of eroded enamel was growing with the increase of the AWSM. This study provides a solution to the resource waste and environmental pollution caused by abandoned abalone shell, and we provides a new method for self-healing remineralization of enamel defects by AWSM and develops a novel dental material for potential clinical dentistry application. - Graphical abstract: In this paper, we successfully isolated the abalone water-soluble matrix (AWSM) with 1.53 wt% abalone water-soluble protein (AWSPro) and 2.04 wt% abalone water-soluble polysaccharide (AWSPs) from abandoned abalone shell, and self-healing biomineralization of tooth defects was successfully achieved in vitro by self-organized. Display Omitted - Highlights: • Provides a solution to the resource waste and environmental pollution caused by abandoned abalone shell. • The abalone shell water-soluble matrix contains protein and polysaccharide. • The abalone water-soluble matrix can efficiently induce remineralization of HAP by self-organized. • Achieved self-healing biomineralization of tooth defects in

  9. Complete mitochondrial genome of Pacific abalone (Haliotis discus hannai) from Korea.

    Science.gov (United States)

    Yang, Eun Chan; Nam, Bo-Hye; Noh, Seung-Jae; Kim, Young-Ok; Kim, Dong-Gyun; Jee, Young Ju; Park, Jun-Hyung; Noh, Jae Hoon; Yoon, Hwan Su

    2015-01-01

    We have sequenced and characterized the complete mitochondrial DNA of an economically and ecologically important Pacific abalone, Haliotis discus hannai (Haliotidae, Gastropoda). The mitogenome of the Pacific abalone is 16,886 nt total length with a 39.6% G+C composition. Thirty-seven genes were identified including 13 protein-coding, 2 rRNA and 22 tRNA genes. We compared the mitogenome of the Pacific abalone to a putative relative species, H. rubra.

  10. Management and Risk Assessment of Sabellid Polychaete Infestations in Abalone Mariculture Facilities

    OpenAIRE

    Kuris, Armand M.

    1996-01-01

    In October 1993, growers at some commercial abalone mariculture facilities (AMFs) in Southern California brought an interesting sabellid polychaete to our attention. The worm infested shells of cultured red abalone, Haliotis rufescens. Heavily infested abalone ceased growth. Their shells exhibited domed teratology often lacking respiratory holes. The worms appeared to bore into the shell. We discovered that all AMFs in California were infested; several were so heavily infest...

  11. Sperm preparation for fertilization

    NARCIS (Netherlands)

    Gadella, B.M.

    2014-01-01

    Description This book contains 19 chapters that discuss theoretical and applied andrology for domestic, zoo and wild animals. Topics include semen and its constituents; sperm production and harvest; determinants of sperm morphology; sperm preparation for fertilization; practical aspects of semen

  12. Sperm proteasomes degrade sperm receptor on the egg zona pellucida during mammalian fertilization.

    Directory of Open Access Journals (Sweden)

    Shawn W Zimmerman

    Full Text Available Despite decades of research, the mechanism by which the fertilizing spermatozoon penetrates the mammalian vitelline membrane, the zona pellucida (ZP remains one of the unexplained fundamental events of human/mammalian development. Evidence has been accumulating in support of the 26S proteasome as a candidate for echinoderm, ascidian and mammalian egg coat lysin. Monitoring ZP protein degradation by sperm during fertilization is nearly impossible because those few spermatozoa that penetrate the ZP leave behind a virtually untraceable residue of degraded proteins. We have overcome this hurdle by designing an experimentally consistent in vitro system in which live boar spermatozoa are co-incubated with ZP-proteins (ZPP solubilized from porcine oocytes. Using this assay, mimicking sperm-egg interactions, we demonstrate that the sperm-borne proteasomes can degrade the sperm receptor protein ZPC. Upon coincubation with motile spermatozoa, the solubilized ZPP, which appear to be ubiquitinated, adhered to sperm acrosomal caps and induced acrosomal exocytosis/formation of the acrosomal shroud. The degradation of the sperm receptor protein ZPC was assessed by Western blotting band-densitometry and proteomics. A nearly identical pattern of sperm receptor degradation, evident already within the first 5 min of coincubation, was observed when the spermatozoa were replaced with the isolated, enzymatically active, sperm-derived proteasomes. ZPC degradation was blocked by proteasomal inhibitors and accelerated by ubiquitin-aldehyde(UBAL, a modified ubiquitin protein that stimulates proteasomal proteolysis. Such a degradation pattern of ZPC is consistent with in vitro fertilization studies, in which proteasomal inhibitors completely blocked fertilization, and UBAL increased fertilization and polyspermy rates. Preincubation of intact zona-enclosed ova with isolated active sperm proteasomes caused digestion, abrasions and loosening of the exposed zonae, and

  13. FORMULASI PAKAN BUATAN DENGAN BAHAN BAKU RUMPUT LAUT UNTUK PERTUMBUHAN ABALON Haliotis squamata

    Directory of Open Access Journals (Sweden)

    I Nyoman Adiasmara Giri

    2016-11-01

    Full Text Available Abalon merupakan salah satu jenis moluska laut yang memiliki potensi ekonomis untuk dikembangkan usaha budidayanya. Teknologi pembenihan abalon Haliotis squamata telah dikembangkan dan berhasil memproduksi benih secara massal. Selain benih, ketersediaan pakan yang sesuai sangat menentukan keberhasilan budidaya pembesaran abalon. Selain makan pakan segar berupa makroalga, abalon juga dapat menerima pakan buatan dengan baik. Penelitian ini bertujuan untuk mendapatkan formula pakan dengan bahan baku tepung rumput laut untuk pertumbuhan H. squamata. Tiga pakan percobaan diformulasi dengan proporsi tepung rumput laut Ulva sp., Gracilaria sp., dan Sargassum sp. yang berbeda dibuat berbentuk remah dengan kandungan protein dan lemak yang sama.  Sebagai kontrol digunakan pakan Gracilaria sp. segar asal tambak.  Sebanyak 16 buah bak beton berukuran 2,0 x 0,5 x 0,5 m3 dengan ketinggian air 25 cm dan dilengkapi dengan aerasi digunakan sebagai wadah percobaan.  Benih abalon hasil pembenihan di hatcheri dengan ukuran bobot 5,30 ± 1,07 g; panjang cangkang 3,35 ± 0,29 cm ditebar dengan kepadatan 388 ekor per bak.  Abalon diberi pakan percobaan 2 kali sehari selama 168 hari. Jumlah pakan disesuaikan dengan respons makan dari abalon. Percobaan dirancang dengan rancangan acak lengkap dengan 4 perlakuan pakan dan setiap perlakuan terdiri atas 4 ulangan.  Hasil percobaan menunjukkan bahwa pertumbuhan abalon yang diberi pakan buatan tidak berbeda nyata, tetapi pakan dengan proporsi tepung Ulva sp. 35% cenderung memberikan pertumbuhan yang lebih baik. Pertumbuhan abalon terbaik diperoleh pada perlakuan kontrol.  Kandungan protein daging abalon dipengaruhi oleh pakan percobaan. Kandungan protein daging abalon yang diberi pakan buatan signifikan lebih tinggi dibandingkan dengan kontrol.

  14. Differentially-Expressed Genes Associated with Faster Growth of the Pacific Abalone, Haliotis discus hannai.

    Science.gov (United States)

    Choi, Mi-Jin; Kim, Gun-Do; Kim, Jong-Myoung; Lim, Han Kyu

    2015-11-18

    The Pacific abalone Haliotis discus hannai is used for commercial aquaculture in Korea. We examined the transcriptome of Pacific abalone Haliotis discus hannai siblings using NGS technology to identify genes associated with high growth rates. Pacific abalones grown for 200 days post-fertilization were divided into small-, medium-, and large-size groups with mean weights of 0.26 ± 0.09 g, 1.43 ± 0.405 g, and 5.24 ± 1.09 g, respectively. RNA isolated from the soft tissues of each group was subjected to RNA sequencing. Approximately 1%-3% of the transcripts were differentially expressed in abalones, depending on the growth rate. RT-PCR was carried out on thirty four genes selected to confirm the relative differences in expression detected by RNA sequencing. Six differentially-expressed genes were identified as associated with faster growth of the Pacific abalone. These include five up-regulated genes (including one specific to females) encoding transcripts homologous to incilarin A, perlucin, transforming growth factor-beta-induced protein immunoglobulin-heavy chain 3 (ig-h3), vitelline envelope zona pellucida domain 4, and defensin, and one down-regulated gene encoding tomoregulin in large abalones. Most of the transcripts were expressed predominantly in the hepatopancreas. The genes identified in this study will lead to development of markers for identification of high-growth-rate abalones and female abalones.

  15. Accurate sperm morphology assessment predicts sperm function.

    Science.gov (United States)

    Abu Hassan Abu, D; Franken, D R; Hoffman, B; Henkel, R

    2012-05-01

    Sperm morphology has been associated with in vitro as well as in vivo fertilisation. The study aimed to evaluate the possible relation between the percentage of spermatozoa with normal morphology and the following sperm functional assays: (i) zona-induced acrosome reaction (ZIAR); (ii) DNA integrity; (iii) chromatin condensation; (iv) sperm apoptosis; and (v) fertilisation rates. Regression analysis was employed to calculate the association between morphology and different functional tests. Normal sperm morphology correlated significantly with the percentages of live acrosome-reacted spermatozoa in the ZIAR (r = 0.518; P sperm apoptosis (r = -0.395; P = 0.0206; n = 34) and necrosis (r = -0.545; P = 0.0009; n = 34). Negative correlations existed between for the acrosome reaction, and DNA integrity, while negative associations were recorded with the percentages of CMA(3) -positive spermatozoa, apoptotic and necrotic spermatozoa. Sperm morphology is related to sperm dysfunction such as poor chromatin condensation, acrosome reaction and DNA integrity. Negative and significant correlations existed between normal sperm morphology and chromatin condensation, the percentage of spermatozoa with abnormal DNA and spermatozoa with apoptotic activity. The authors do not regard sperm morphology as the only test for the diagnosis of male fertility, but sperm morphology can serve as a valuable indicator of underlying dysfunction. © 2011 Blackwell Verlag GmbH.

  16. Expanding lysine industry: industrial biomanufacturing of lysine and its derivatives.

    Science.gov (United States)

    Cheng, Jie; Chen, Peng; Song, Andong; Wang, Dan; Wang, Qinhong

    2018-04-13

    L-Lysine is widely used as a nutrition supplement in feed, food, and beverage industries as well as a chemical intermediate. At present, great efforts are made to further decrease the cost of lysine to make it more competitive in the markets. Furthermore, lysine also shows potential as a feedstock to produce other high-value chemicals for active pharmaceutical ingredients, drugs, or materials. In this review, the current biomanufacturing of lysine is first presented. Second, the production of novel derivatives from lysine is discussed. Some chemicals like L-pipecolic acid, cadaverine, and 5-aminovalerate already have been obtained at a lab scale. Others like 6-aminocaproic acid, valerolactam, and caprolactam could be produced through a biological and chemical coupling pathway or be synthesized by a hypothetical pathway. This review demonstrates an active and expansive lysine industry, and these green biomanufacturing strategies could also be applied to enhance the competitiveness of other amino acid industry.

  17. A comparison of DNA compaction by arginine and lysine peptides: A physical basis for arginine rich protamines

    OpenAIRE

    DeRouchey, Jason; Hoover, Brandon; Rau, Donald C

    2013-01-01

    Protamines are small, highly positively charged peptides used to package DNA to very high densities in sperm nuclei. Tight DNA packing is considered essential to minimize DNA damage by mutagens and reactive oxidizing species. A striking and general feature of protamines is the almost exclusive use of arginine over lysine for the positive charge to neutralize DNA. We have investigated whether this preference for arginine might arise from a difference in DNA condensation by arginine and lysine ...

  18. Reduced disease in black abalone following mass mortality: Phage therapy and natural selection

    Science.gov (United States)

    VanBlaricom, Glenn R.

    2014-01-01

    Black abalone, Haliotis cracherodii, populations along the NE Pacific ocean have declined due to the rickettsial disease withering syndrome (WS). Natural recovery on San Nicolas Island (SNI) of Southern California suggested the development of resistance in island populations. Experimental challenges in one treatment demonstrated that progeny of disease-selected black abalone from SNI survived better than did those from naïve black abalone from Carmel Point in mainland coastal central California. Unexpectedly, the presence of a newly observed bacteriophage infecting the WS rickettsia (WS-RLO) had strong effects on the survival of infected abalone. Specifically, presence of phage-infected RLO (RLOv) reduced the host response to infection, RLO infection loads, and associated mortality. These data suggest that the black abalone: WS-RLO relationship is evolving through dual host mechanisms of resistance to RLO infection in the digestive gland via tolerance to infection in the primary target tissue (the post-esophagus) coupled with reduced pathogenicity of the WS-RLO by phage infection, which effectively reduces the infection load in the primary target tissue by half. Sea surface temperature patterns off southern California, associated with a recent hiatus in global-scale ocean warming, do not appear to be a sufficient explanation for survival patterns in SNI black abalone. These data highlight the potential for natural recovery of abalone populations over time and that further understanding of mechanisms governing host–parasite relationships will better enable us to manage declining populations.

  19. Sperm Surface Proteomics 2

    NARCIS (Netherlands)

    Brewis, I.A.; Gadella, B.M.|info:eu-repo/dai/nl/115389873

    2017-01-01

    This contribution will focus exclusively on the total (global) protein composition (the proteome) of the sperm surface. Immune responses directed towards sperm surface proteins may cause infertility since functionally intact sperm are under immune attack. The immune attack can be achieved directly

  20. A comparison of DNA compaction by arginine and lysine peptides: a physical basis for arginine rich protamines.

    Science.gov (United States)

    DeRouchey, Jason; Hoover, Brandon; Rau, Donald C

    2013-04-30

    Protamines are small, highly positively charged peptides used to package DNA at very high densities in sperm nuclei. Tight DNA packing is considered essential for the minimization of DNA damage by mutagens and reactive oxidizing species. A striking and general feature of protamines is the almost exclusive use of arginine over lysine for the positive charge to neutralize DNA. We have investigated whether this preference for arginine might arise from a difference in DNA condensation by arginine and lysine peptides. The forces underlying DNA compaction by arginine, lysine, and ornithine peptides are measured using the osmotic stress technique coupled with X-ray scattering. The equilibrium spacings between DNA helices condensed by lysine and ornithine peptides are significantly larger than the interhelical distances with comparable arginine peptides. The DNA surface-to-surface separation, for example, is some 50% larger with polylysine than with polyarginine. DNA packing by lysine rich peptides in sperm nuclei would allow much greater accessibility to small molecules that could damage DNA. The larger spacing with lysine peptides is caused by both a weaker attraction and a stronger short-range repulsion relative to that of the arginine peptides. A previously proposed model for binding of polyarginine and protamine to DNA provides a convenient framework for understanding the differences between the ability of lysine and arginine peptides to assemble DNA.

  1. Antibiotic resistance monitoring in Vibrio spp. isolated from rearing environment and intestines of abalone Haliotis diversicolor.

    Science.gov (United States)

    Wang, R X; Wang, J Y; Sun, Y C; B L Yang; A L Wang

    2015-12-30

    546 Vibrio isolates from rearing seawater (292 strains) and intestines of abalone (254 strains) were tested to ten antibiotics using Kirby-Bauer diffusion method. Resistant rates of abalone-derived Vibrio isolates to chloramphenicol (C), enrofloxacin (ENX) and norfloxacin (NOR) were 40%) to kanamycin (KNA), furazolidone (F), tetracycline (TE), gentamicin (GM) and rifampin (RA). 332 isolates from seawater (n=258) and abalone (n=74) were resistant to more than three antibiotics. Peaked resistant rates of seawater-derived isolates to multiple antibiotics were overlapped in May and August. Statistical analysis showed that pH had an important effect on resistant rates of abalone-derived Vibrio isolates to RA, NOR, and ENX. Salinity and dissolved oxygen were negatively correlated with resistant rates of seawater-derived Vibrio isolates to KNA, RA, and PG. Copyright © 2015 Elsevier Ltd. All rights reserved.

  2. A family of microbial lysine transporter polypeptides

    DEFF Research Database (Denmark)

    2017-01-01

    The present invention provides a genetically modified microbial cell for production of lysine, comprising a transgene encoding a polypeptide capable of exporting lysine from the cell. The genetically modified microbial cell for production of lysine may be further characterized by genetic modifica......The present invention provides a genetically modified microbial cell for production of lysine, comprising a transgene encoding a polypeptide capable of exporting lysine from the cell. The genetically modified microbial cell for production of lysine may be further characterized by genetic...... a novel family of lysine transporter polypeptides; and the use of said polypeptide to enhance production of extracellular lysine in a microbial cell....

  3. Daily sperm production

    DEFF Research Database (Denmark)

    Kyjovska, Zdenka Orabi; Boisen, Anne Mette Zenner; Jackson, Petra

    2013-01-01

    instillation with Printex90. Body and testicle weight, sperm content per g testicular parenchyma and daily sperm production (DSP) were assessed. The protocol for assessment of DSP was optimized for application in mice (C57BL/6J) and the influence of different parameters was studied. Maternal particulate...... exposure did not affect DSP statistically significantly in the F1 generation, although TiO2 tended to reduce sperm counts. Overall, time-to-first F2 litter increased with decreasing sperm production. There was no effect on sperm production in the F2 generation originating after TiO2 exposure. F2 offspring......, whose fathers were prenatally exposed to Printex90, showed lowered sperm production. Furthermore, we report statistically significant differences in sperm production between mouse strains....

  4. Species Profiles. Life Histories and Environmental Requirements of Coastal Fishes and Invertebrates (Pacific Southwest). BLACK, GREEN, and RED ABALONES.

    Science.gov (United States)

    1985-03-01

    abalone landings abalones grow fast. Under optimal and ex-vissel value in California, conditions in a laboratory culture , 1965-1982 some juvenile red...Lithothamnion sp. Hymenena sp. Plocamium coccineum Tr i daea sp. Rhodymenia sp. Lithothamnion sp. Porphyra sp. Prionitis sp. Rhodymenia sp...Fecundacion artifi- Abalones, shells, and sea otters: cal y dessarrollo embrionario de dynamics of prey populations in Haliotis fulgens Philippi, 1845

  5. Status of abalone fishery and experiential mariculture as a resource conservation strategy in Carot, Anda, Pangasinan

    Directory of Open Access Journals (Sweden)

    Emmanuel C. Capinpin, Jr.

    2013-06-01

    Full Text Available The study described the abalone f ishery in Carot, Anda, Pangasinan to develop mariculture and to reseed a part of the harvest as a resource conservation strategy. The abalone fishery of Anda is artisanal or smallscale, typif ied by f ishers gleaning or free-diving on shallow rocky areas which are the typical habitat of abalone. Low densities of 1.67 to 8 individuals per 250 m2 were observed. Local f ishers have knowledge of productive f ishing areas. Hence, cage culture of abalone in these areas could be a viable resource conservation strategy as they serve as reproductive reserves to supply larvae for continued productivity of the f ishing grounds. Abalone mariculture following the Farmer Field School (FFS concept was explored to address both resource management and economic needs. As a resource enhancement activity, mariculture guarantees that cultured abalone are allowed to grow to maturity before harvested, while some are retained to restock a marine sanctuary. Sincemariculture makes possible the aggregation of individuals, the probability that fertilization would take place is increased. As supplemental source of livelihood, abalone is a high value commodity and its culture can help augment the income of f ishers. Small abalone (3-4 cm can be cultured further for 3-4 months to increase their size and weight. Mariculture should be done from November to May to avoid the rainy season and improve survivorship. The experiential activity was successful because it became a means for the f ishers to experienceresource management. Under the FFS, the researcher became a facilitator and mentored the cooperators in learning from their experience. The lessons sharpened the f ishers’ skills in observation, problem-solving, decision-making, and critical thinking. This enabled them to gain an appreciation of their resource.

  6. Using Local Ecological Knowledge and Environmental Education in Resource Management of Abalone in Carot, Anda, Pangasinan

    Directory of Open Access Journals (Sweden)

    Emmanuel C. Capinpin, Jr.

    2012-06-01

    Full Text Available The objectives of the present study were to (1 determine the local ecological knowledge (LEK of abalone gatherers through interviews and mentoring, and assess the correspondence between scientific information and LEK, so that areas where local knowledge may be most useful in resource management could be identified, and (2 to empower selected gatherers/farmers with knowledge and technical skills through environmental education to help develop or build their capacity to become sustainable resource managers. The LEK of abalone fishers was determined using three complementary approaches – group interview, individual interview, and mentoring sessions. Local fishers possess a wealth of knowledge about the interactions of species gained through many years of observations, and this knowledge may be useful in guiding biologists in ecological restoration or management regimes. Additionally, the fishers’ LEK, validated by modern scientific ecological findings, could be a source of important and effective ideas in resource management. The knowledge of the abalone gatherers about important abalone fishing grounds should help in pinpointing critical areas that need to be managed. Abalone mariculture in cages should be set up in these areas to routinely create dense breeding populations which can help in enhancing recovery and in providing fishers with a source of additional income. The continued enforcement of marine protected areas and the periodic release or reseeding of abalone in sanctuaries could also be considered viable resource management options. Other recommendations for resource management based on gathered local knowledge and lessons learned from the environmental education (EE seminars are also presented.

  7. Potential mechanisms of phthalate ester embryotoxicity in the abalone Haliotis diversicolor supertexta

    Energy Technology Data Exchange (ETDEWEB)

    Zhou Jin [L-304, Life Sciences Division, Graduate School at Shenzhen, Tsinghua University, Shenzhen University Town, Xili, Shenzhen City 518055 (China); Cai Zhonghua, E-mail: caizh@sz.tsinghua.edu.cn [L-304, Life Sciences Division, Graduate School at Shenzhen, Tsinghua University, Shenzhen University Town, Xili, Shenzhen City 518055 (China); Key Laboratory of Aquatic-Ecology, Tianjin Agricultural University, Lishui Road 112, Tianjin 300384 (China); Xing Kezhi [Key Laboratory of Aquatic-Ecology, Tianjin Agricultural University, Lishui Road 112, Tianjin 300384 (China)

    2011-05-15

    The effects and associated toxicological mechanisms of five phthalate esters (PAEs) on abalone embryonic development were investigated by exposing the embryos to a range of PAEs concentrations (0.05, 0.2, 2 and 10 {mu}g/mL). The results showed that PAEs could significantly reduce embryo hatchability, increase developmental malformations, and suppress the metamorphosis of abalone larvae. The possible toxicological mechanisms of PAEs to abalone embryos included, affecting the Na{sup +}-K{sup +}-pump and Ca{sup 2+}-Mg{sup 2+}-pump activities, altering the peroxidase (POD) level and the malondialdehyde (MDA) production, damaging the extraembryonic membranes structure, as well as disrupting endocrine-related genes (gpx, cyp3a, and 17{beta}-hsd 12) expression properties. Taken together, this work showed that PAEs adversely affected the embryonic ontogeny of abalone. The abilities of PAEs affecting the osmoregulation, inducing oxidative stress, damaging embryo envelope structure, and causing physiological homeostasis disorder, are likely to be a part of the common mechanisms responsible for their embryonic toxicity. - Highlights: > PAEs affected abalone hatchability, morphogenesis and metamorphosis behavior. > The toxicity of the five PAEs to embryogenesis was ranked as DBP > DEP > DMP > DOP > DEHP. > The osmoregulation disorder and oxidative damage are the potential mechanisms. - Potential mechanisms of PAEs on abalone embryogenesis are osmoregulation disorder, oxidative damage and physiological dysfunction.

  8. Sperm preparation for fertilization

    OpenAIRE

    Gadella, B.M.

    2014-01-01

    Description This book contains 19 chapters that discuss theoretical and applied andrology for domestic, zoo and wild animals. Topics include semen and its constituents; sperm production and harvest; determinants of sperm morphology; sperm preparation for fertilization; practical aspects of semen cryopreservation; evaluation of semen in the andrology laboratory; genetic aspects of male reproduction; emerging techniques and future development of semen evaluation and handling and applied androlo...

  9. Rheotaxis guides mammalian sperm

    Science.gov (United States)

    Miki, Kiyoshi; Clapham, David E

    2013-01-01

    Background In sea urchins, spermatozoan motility is altered by chemotactic peptides, giving rise to the assumption that mammalian eggs also emit chemotactic agents that guide spermatozoa through the female reproductive tract to the mature oocyte. Mammalian spermatozoa indeed undergo complex adaptations within the female (the process of capacitation) that are initiated by agents ranging from pH to progesterone, but these factors are not necessarily taxic. Currently, chemotaxis, thermotaxis, and rheotaxis have not been definitively established in mammals. Results Here, we show that positive rheotaxis, the ability of organisms to orient and swim against the flow of surrounding fluid, is a major taxic factor for mouse and human sperm. This flow is generated within 4 hours of sexual stimulation and coitus in female mice; prolactin-triggered oviductal fluid secretion clears the oviduct of debris, lowers viscosity, and generates the stream that guides sperm migration in the oviduct. Rheotaxic movement is demonstrated in capacitated and uncapacitated spermatozoa in low and high viscosity medium. Finally, we show that a unique sperm motion we quantify using the sperm head's rolling rate reflects sperm rotation that generates essential force for positioning the sperm in the stream. Rotation requires CatSper channels, presumably by enabling Ca2+ influx. Conclusions We propose that rheotaxis is a major determinant of sperm guidance over long distances in the mammalian female reproductive tract. Coitus induces fluid flow to guide sperm in the oviduct. Sperm rheotaxis requires rotational motion during CatSper channel-dependent hyperactivated motility. PMID:23453951

  10. Effect of abalone farming on sediment geochemistry in the Shallow Sea near Wando, South Korea

    Science.gov (United States)

    Kang, Jeongwon; Lee, Yeon Gyu; Jeong, Da Un; Lee, Jung Sick; Choi, Yang Ho; Shin, Yun Kyung

    2015-12-01

    Wando County has grown up to 93% of the total abalone produced in South Korea since the late 1990s; however, this production has been decreasing in recent years. The objectives of this study were to understand the potential contamination risks of abalone farming and to examine the influence of intensive abalone farming on sediment quality by analyzing grain-size composition, organic matter (total organic carbon (TOC), total nitrogen (TN), total sulfur (TS)) and heavy metal content, pH, and 210Pb geochronology. The results of organic matter analysis from surface and core sediment (length: 64 cm) showed that the area around the abalone farm had oxic marine-to-brackish conditions, but that the area directly below an abalone cage (location 7) had reductive conditions, with a C/S ratio of ~2. The average TN levels in the surface and core sediments were 0.25% and 0.29%, respectively, and this was predominantly due to the use of seaweed for feed. The low sediment pH (surface, 7.23; core, 7.04), indicates that acidification of the bottom sediment has gradually increased since the initiation of abalone farming and is likely due to the continuous accumulation of uneaten feed and feces. Heavy metal pollution was not apparent based on the examination of EF and Igeo, although the excess metal flux of Ni, Pb, Cu, Co, As, and Cd increased toward surface of the sediment core. These sediment changes may be caused by the rapid accumulation (sedimentation rate: 1.45 cm/year) of sludge discharged from the abalone farm and may be controlled by tidal currents, physiography, water depth, and tidal ranges.

  11. Genetic differentiation between fake abalone and genuine Haliotis species using the forensically informative nucleotide sequencing (FINS) method.

    Science.gov (United States)

    Ha, Wai Y; Reid, David G; Kam, Wan L; Lau, Yuk Y; Sham, Wing C; Tam, Silvia Y K; Sin, Della W M; Mok, Chuen S

    2011-05-25

    Abalones ( Haliotis species) are a popular delicacy and commonly preserved in dried form either whole or in slices or small pieces for consumption in Asian countries. Driven by the huge profit from trading abalones, dishonest traders may substitute other molluscan species for processed abalone, of which the morphological characteristics are frequently lost in the processed form. For protection of consumer rights and law enforcement against fraud, there is a need for an effective methodology to differentiate between fake and genuine abalone. This paper describes a method (validated according to the international forensic guidelines provided by SWGDAM) for the identification of fake abalone species using forensically informative nucleotide sequence (FINS) analysis. A study of the local market revealed that many claimed "abalone slice" samples on sale are not genuine. The fake abalone samples were found to be either volutids of the genus Cymbium (93%) or the muricid Concholepas concholepas (7%). This is the first report of Cymbium species being used for the preparation and sale as "abalone" in dried sliced form in Hong Kong.

  12. Epipodial Tentacle Gene Expression and Predetermined Resilience to Summer Mortality in the Commercially Important Greenlip Abalone, Haliotis laevigata.

    Science.gov (United States)

    Shiel, Brett P; Hall, Nathan E; Cooke, Ira R; Robinson, Nicholas A; Strugnell, Jan M

    2017-04-01

    "Summer mortality" is a phenomenon that occurs during warm water temperature spikes that results in the mass mortality of many ecologically and economically important mollusks such as abalone. This study aimed to determine whether the baseline gene expression of abalone before a laboratory-induced summer mortality event was associated with resilience to summer mortality. Tentacle transcriptomes of 35 greenlip abalone (Haliotis laevigata) were sequenced prior to the animals being exposed to an increase in water temperature-simulating conditions which have previously resulted in summer mortality. Abalone derived from three source locations with different environmental conditions were categorized as susceptible or resistant to summer mortality depending on whether they died or survived after the water temperature was increased. We detected two genes showing significantly higher expression in resilient abalone relative to susceptible abalone prior to the laboratory-induced summer mortality event. One of these genes was annotated through the NCBI non-redundant protein database using BLASTX to an anemone (Exaiptasia pallida) Transposon Ty3-G Gag Pol polyprotein. Distinct gene expression signatures were also found between resilient and susceptible abalone depending on the population origin, which may suggest divergence in local adaptation mechanisms for resilience. Many of these genes have been suggested to be involved in antioxidant and immune-related functions. The identification of these genes and their functional roles have enhanced our understanding of processes that may contribute to summer mortality in abalone. Our study supports the hypothesis that prestress gene expression signatures are indicative of the likelihood of summer mortality.

  13. KERAGAAN PERTUMBUHAN DAN REPRODUKSI ABALON Haliotis squamata Reeve (1846 TURUNAN KETIGA

    Directory of Open Access Journals (Sweden)

    Gusti Ngurah Permana

    2018-01-01

    Full Text Available Pengamatan pertumbuhan dan reproduksi abalon Haliotis squamata dilakukan di hatcheri Balai Besar Riset Budidaya Laut dan Penyuluhan Perikanan (BBRBLPP Gondol, Bali. Tujuan dari penelitian ini untuk memperoleh informasi tentang keragaan pertumbuhan dan performansi reproduksi abalon turunan ketiga. Induk H. squamata turunan kedua hasil seleksi yang digunakan untuk menghasilkan benih turunan ketiga mempunyai ukuran panjang cangkang 6,5-7,0 cm. Benih dipelihara dalam bak beton berukuran 2,5 m x 1,2 m x 1,0 m yang diberikan feeding plate sebagai substrat penempelan dan dilengkapi dengan sistem aerasi dan sistem air mengalir. Pakan yang diberikan pada awal pemeliharaan adalah diatom jenis Nitzschia sp. dan Melosira sp. yang telah ditumbuhkan terlebih dahulu pada feeding plate sebelum penebaran benih. Benih F-3 dipelihara sampai menjadi calon induk untuk diamati perkembangan reproduksinya. Pengambilan sampel pertumbuhan dilakukan setiap 10 hari. Pengamatan reproduksi dilakukan pada saat abalon mulai tumbuh gonad sampai matang gonad stadia-III. Hasil penelitian menunjukkan bahwa pertumbuhan abalon sangat dipengaruhi ketersediaan pakan pada plate terutama pada hari ke-50. Proporsi jantan-betina abalon F-3 (3,3:1 meningkat dibandingkan dengan F-0 dari alam (2,5:1 menunjukkan ketidakseimbangan jumlah individu yang dapat disebabkan oleh tekanan seleksi. Abalon turunan ketiga pada umur 16 bulan mulai matang gonad dan dapat digunakan sebagai induk untuk pemijahan. Observation on the growth and reproduction development of Haliotis squamata had been undertaken in the hatchery of the Institute for Mariculture Research and Development (IMRAD Gondol, Bali. The research was aimed to study of the growth and reproduction performance of filial-3 abalone in supporting seed production in hatchery. Larvae were obtained from natural spawning of filial-2 abalone broodstock with the length shell of 6.5-7.0 cm in the hatchery. Larvae were reared in 2.5 m x 1.2 m x 1.0 m

  14. Evaluating potential conservation conflicts between two listed species: sea otters and black abalone.

    Science.gov (United States)

    Raimondi, Peter; Jurgens, Laura J; Tinker, M Tim

    2015-11-01

    Population consequences of endangered species interacting as predators and prey have been considered theoretically and legally, but rarely investigated in the field. We examined relationships between spatially variable populations of a predator, the California sea otter, Enhydra lutris nereis, and a prey species, the black abalone, Haliotis cracherodii. Both species are federally listed under the Endangered Species Act and co-occur along the coast of California. We compared the local abundance and habitat distribution of black abalone at 12 sites with varying densities of sea otters. All of the populations of abalone we examined were in the geographic area currently unaffected by withering disease, which has decimated populations south of the study area. Surprisingly, our findings indicate that sea otter density is positively associated with increased black abalone density. The presence of sea otters also correlated with a shift in black abalone to habitat conferring greater refuge, which could decrease illegal human harvest. These results highlight the need for a multi-species approach to conservation management of the two species, and demonstrate the importance of using field-collected data rather than simple trophic assumptions to understand relationships between jointly vulnerable predator and prey populations.

  15. Preparation, characterisation and use for antioxidant oligosaccharides of a cellulase from abalone (Haliotis discus hannai) viscera.

    Science.gov (United States)

    Tao, Zhi-Peng; Sun, Le-Chang; Qiu, Xu-Jian; Cai, Qiu-Feng; Liu, Guang-Ming; Su, Wen-Jin; Cao, Min-Jie

    2016-07-01

    In China, abalone (Haliotis discus hannai) production is growing annually. During industrial processing, the viscera, which are abundant of cellulase, are usually discarded or processed into low-value feedstuff. Thus, it is of interest to obtain cellulase from abalone viscera and investigate its application for preparation of functional oligosaccharides. A cellulase was purified from the hepatopancreas of abalone by ammonium sulfate precipitation and two-steps column chromatography. The molecular weight of the cellulase was 45 kDa on SDS-PAGE. Peptide mass fingerprinting analysis yielded 103 amino acid residues, which were identical to cellulases from other species of abalone. Substrate specificity analysis indicated that the cellulase is an endo-1,4-β-glucanase. Hydrolysis of seaweed Porphyra haitanensis polysaccharides by the enzyme produced oligosaccharides with degree of polymerisation of two to four, whose monosaccharide composition was 58% galactose, 4% glucose and 38% xylose. The oligosaccharides revealed 2,2'-diphenyl-1-picrylhydrazyl free radical as well as hydrogen peroxide scavenging activity. It is feasible and meaningful to utilise cellulase from the viscera of abalone for preparation of functional oligosaccharides. © 2015 Society of Chemical Industry. © 2015 Society of Chemical Industry.

  16. Evaluating potential conservation conflicts between two listed species: Sea otters and black abalone

    Science.gov (United States)

    Raimondi, Peter T.; Jurgens, Laura J.; Tinker, M. Tim

    2015-01-01

    Population consequences of endangered species interacting as predators and prey have been considered theoretically and legally, but rarely investigated in the field. We examined relationships between spatially variable populations of a predator, the California sea otter, Enhydra lutris nereis, and a prey species, the black abalone, Haliotis cracherodii. Both species are federally listed under the Endangered Species Act and co-occur along the coast of California. We compared the local abundance and habitat distribution of black abalone at 12 sites with varying densities of sea otters. All of the populations of abalone we examined were in the geographic area currently unaffected by withering disease, which has decimated populations south of the study area. Surprisingly, our findings indicate that sea otter density is positively associated with increased black abalone density. The presence of sea otters also correlated with a shift in black abalone to habitat conferring greater refuge, which could decrease illegal human harvest. These results highlight the need for a multi-species approach to conservation management of the two species, and demonstrate the importance of using field-collected data rather than simple trophic assumptions to understand relationships between jointly vulnerable predator and prey populations.

  17. KERAGAAN PERTUMBUHAN DAN VARIASI GENETIK ABALON Haliotis squamata Reeve (1846 HASIL SELEKSI F-1

    Directory of Open Access Journals (Sweden)

    Gusti Ngurah Permana

    2015-12-01

    Full Text Available Produksi benih abalon Haliotis squamata skala massal di hatcheri telah berhasil dilakukan di Balai Besar Penelitian dan Pengembangan Budidaya Laut Gondol, Bali. Permasalahan utama dalam budidaya abalon adalah pertumbuhan yang lambat. Keadaan tersebut diduga karena pengaruh faktor genetik dan lingkungan. Penelitian ini bertujuan mengetahui keragaan pertumbuhan dan variasi genetik abalon tumbuh cepat hasil seleksi individu. Hasil penelitian ini diketahui bahwa pembentukan populasi F-1 mempunyai pertumbuhan yang lebih baik dengan F-1 kontrol. Peningkatan bobot yang dicapai 22,15 g atau 17,93% lebih baik dibandingkan F-1 kontrol. Keragaman genetik F-1 terseleksi yang ditunjukkan dari nilai heterozigositas adalah (Ho. 0,023 terjadi penurunan 21,7% jika dibandingkan F-0. Hal ini dapat terjadi karena hilangnya beberapa allele dalam proses seleksi. Terdapat hubungan antara jumlah heterozigot pada lokus tertentu dengan pertumbuhan abalon. Hasil ini diharapkan dapat mendukung upaya meningkatkan produksi benih yang mempunyai performa fenotipe dan genotipe unggul sehingga dapat mendukung kegiatan budidaya abalon yang berkelanjutan.

  18. Involvement of Antizyme Characterized from the Small Abalone Haliotis diversicolor in Gonadal Development.

    Directory of Open Access Journals (Sweden)

    Wei-Dong Li

    Full Text Available The small abalone Haliotis diversicolor is an economically important mollusk that is widely cultivated in Southern China. Gonad precocity may affect the aquaculture of small abalone. Polyamines, which are small cationic molecules essential for cellular proliferation, may affect gonadal development. Ornithine decarboxylase (ODC and antizyme (AZ are essential elements of a feedback circuit that regulates cellular polyamines. This paper presents the molecular cloning and characterization of AZ from small abalone. Sequence analysis showed that the cDNA sequence of H. diversicolor AZ (HdiODCAZ consisted of two overlapping open reading frames (ORFs and conformed to the +1 frameshift property of the frame. Thin Layer chromatography (TLC analysis suggested that the expressed protein encoded by +1 ORF2 was the functional AZ that targets ODC to 26S proteasome degradation. The result demonstrated that the expression level of AZ was higher than that of ODC in the ovary of small abalone. In addition, the expression profiles of ODC and AZ at the different development stages of the ovary indicated that these two genes might be involved in the gonadal development of small abalone.

  19. Sperm storage potential and daily sperm production of brown male ...

    African Journals Online (AJOL)

    Sperm storage potential and daily sperm production of brown male Japanese quails for three different physiological age groups. ... positively correlated (r=0.91 P<0.01). This study revealed that the brown male Japanese quail has its peak reproduction potential at puberty. Keywords: Quail sperm storage, sperm production ...

  20. Sperm form and function in the absence of sperm competition.

    Science.gov (United States)

    van der Horst, Gerhard; Maree, Liana

    2014-03-01

    Sperm competition is a post-copulatory, sexual selection force that, together with phylogeny and fertilization mode, has been regarded as one of the main factors explaining the diversity in sperm size across species. This universal sperm selection mechanism favors traits that enhance a male's fertilizing ability and paternity success. Surprisingly, however, sperm characteristics and semen quality in monogamous species, with low risk of sperm competition, have barely received any attention. In this review, we consider sperm competition and monogamy as two ends of the selective spectrum, and discuss its effect on sperm structure and function. We address the issue of a lack of sperm competition by comparing sperm traits of essentially monogamous species--their largely degenerative sperm features and high degree of polymorphisms could be norms for monogamous species. Further, the level of sperm competition in humans is discussed by comparing its mating strategy, relative testis size, and sperm traits to other primate species. In terms of sperm concentration, sperm swimming speed, and sperm morphology, humans seem to be closer aligned to the low-risk sperm competition situation in gorillas than to promiscuous chimpanzees. © 2013 Wiley Periodicals, Inc.

  1. Isolation, identification, and antioxidant activity of polysaccharides from the shell of abalone (Haliotis discus hannai Ino).

    Science.gov (United States)

    Wang, Z L; Liang, H B; Guo, W; Peng, Z F; Chen, J D; Zhang, Q Q

    2014-07-04

    In this study, two antioxidative substances, a homogeneous polysaccharide [abalone shell polysaccharide (ASP-1), corresponding to the first peak by size exclusion chromatography] and a non-polysaccharide compound [abalone shell compound (ACS-2), corresponding to the second peak by size exclusion chromatography], were extracted from the abalone (Haliotis discus hannai Ino) shell. We primarily focused on the investigation of ASP-1. As a heteropolysaccharide, ASP-1 is comprised of 9.3% uronic acid and 86.4% saccharide, the latter including mannose, ribose, rhamnose, glucose, galactose, arabinose, and two unknown monosaccharides, NO1 and NO2, with a mass ratio of 9.5:10.1:2.2:18.2:21.8:5.5:16.5:16.2. The antioxidant activity assays indicated that 5.0 mg/mL ASP-1 has significant scavenging effects on superoxide radicals (86.2%) compared to the positive control of ascorbic acid (95.6%).

  2. Biokinetic behavior of Tc in the red abalone, Haliotis rufescens: a reassessment

    International Nuclear Information System (INIS)

    Beasley, T.M.; Lorz, H.V.; Gonor, J.J.

    1981-01-01

    The biokinetic behavior of /sup 95m/Tc in the red abalone, Haliotis rufescens, is reviewed in light of recent experiments on other molluscs. Additional experimentation has confirmed that, when uptake is directly from labeled seawater, abalone exhibit concentration factors in excess of 100. Bivalve molluscs under the same experimental conditions have concentration factors that do not exceed 2. However, uptake and loss kinetics cannot be described by a single compartment model as had been previously advanced. Assimilation of /sup 95m/Tc by the abalone following a single feeding of labeled macroalga, Nereocystis pyrifera, is high (approx. 45%) and loss kinetics are similar to those observed following direct uptake from seawater

  3. Lysine requirements of growing emus.

    Science.gov (United States)

    Mannion, P F; Kent, P B; Barram, K M; Trappett, P C; Blight, G W; Sales, J

    1999-05-01

    1. The lysine requirement of growing emus between 23 and 65 d of age was determined according to growth response variables. 2. The optimal lysine requirement of emus was found to be 0.83 and 0.90 g/MJ ME for growth rate and gain:food ratio respectively. These findings are in accordance with the recommended value of 0.80 g/MJ ME, but is lower than the recommended value for ostriches (1.02 g/MJ ME) and higher than determined values for broilers (0.75 g/MJ ME) of the same age range.

  4. Transcriptional responses of metallothionein gene to different stress factors in Pacific abalone (Haliotis discus hannai).

    Science.gov (United States)

    Lee, Sang Yoon; Nam, Yoon Kwon

    2016-11-01

    A novel metallothionein (MT) gene from the Pacific abalone H. discus hannai was characterized and its mRNA expression patterns (tissue distribution, developmental expression and differential expression in responsive to various in vivo stimulatory treatments) were examined. Abalone MT shares conserved structural features with previously known gastropod orthologs at both genomic (i.e., tripartite organization) and amino acid (conserved Cys motifs) levels. The 5'-flanking regulatory region of abalone MT gene displayed various transcription factor binding motifs particularly including ones related with metal regulation and stress/immune responses. Tissue distribution and basal expression patterns of MT mRNAs indicated a potential association between ovarian MT expression and sexual maturation. Developmental expression pattern suggested the maternal contribution of MT mRNAs to embryonic and early larval developments. Abalone MT mRNAs could be significantly induced by various heavy metals in different tissues (gill, hepatopancreas, muscle and hemocyte) in a tissue- and/or metal-dependent fashion. In addition, the abalone MT gene was highly modulated in responsive to other non-metal, stimulatory treatments such as immune challenge (LPS, polyI:C and bacterial injections), hypoxia (decrease from normoxia 8 ppm-2 ppm), thermal elevation (increase from 20 °C to 30 °C), and xenobiotic exposure (250 ppb of 17α-ethynylestradiol and 0.25 ppb of 2,3,7,8-tetrachlorodibenzodioxin) where differential expression patterns were toward either up- or down-regulation depending on types of stimulations and tissues examined. Taken together, our results highlight that MT is a multifunctional effector playing in wide criteria of cellular pathways especially associated with development and stress responses in this abalone species. Copyright © 2016 Elsevier Ltd. All rights reserved.

  5. Genomic Characterization of a Novel Phage Found in Black Abalone (Haliotis cracherodii) Infected with Withering Syndrome

    Science.gov (United States)

    Closek, C. J.; Langevin, S.; Burge, C. A.; Crosson, L.; White, S.; Friedman, C. S.

    2016-02-01

    Withering syndrome (WS), caused by the bacterium Candidatus Xenohaliotis californiensis, a Rickettsia-like organism (RLO), infects many species of abalone. Black abalone (Haliotis cracherodii), one of two endangered species of abalone, has experienced high population losses along the California coast due to WS. Recently, we observed reduced pathogenicity and mortality events in RLO-infected abalone when a novel bacteriophage (phage) was also present. To better understand phage-bacterium dynamics and develop more informative diagnostic tools, we sequenced the genome of the novel phage associated with the RLO responsible for WS. Metagenomic sequencing libraries were prepared with extracted genomic DNA from two experimentally infected H. cracherodii and phage sequences were enriched using hydroxyapatite chromatography normalization. Normalized libraries were individually barcoded and sequenced with Illumina MiSeq. Raw sequence reads were processed using VIrominer and de novo assembly produced one single phage-like contig (35.7Kb) from the experimentally infected abalone. This highly divergent genome had closest homology with a virus associated with abalone shriveling syndrome (SS). Of the 34 predicted ORFs, overlapping homology with the SS virus ranged from 20-72%, demonstrating the phage sequenced is genetically distinct from any known phage. The phage-like sequences represented a significant portion of the total reads sequenced ( 2 million of the 12 million paired-end reads; 17%) and we obtained 94,000X coverage across the novel phage genome. Beyond characterization of this novel phage, which appears to reduce pathogenicity of the RLO, the genome enabled us to develop quantitative PCR and in situ hybridization assays as diagnostic tools. These tools allow us to detect and quantify this phage in the endangered H. cracherodii.

  6. Low Sperm Count

    Science.gov (United States)

    ... problem such as an inherited chromosomal abnormality, a hormonal imbalance, dilated testicular veins or a condition that blocks ... Elevated temperatures impair sperm production and function. Although studies are limited and are inconclusive, frequent use of ...

  7. Abalone visceral extract inhibit tumor growth and metastasis by modulating Cox-2 levels and CD8+ T cell activity

    Directory of Open Access Journals (Sweden)

    II Kim Jae

    2010-10-01

    Full Text Available Abstract Background Abalone has long been used as a valuable food source in East Asian countries. Although the nutritional importance of abalone has been reported through in vitro and in vivo studies, there is little evidence about the potential anti-tumor effects of abalone visceral extract. The aim of the present study is to examine anti-tumor efficacy of abalone visceral extract and to elucidate its working mechanism. Methods In the present study, we used breast cancer model using BALB/c mouse-derived 4T1 mammary carcinoma and investigated the effect of abalone visceral extract on tumor development. Inhibitory effect against tumor metastasis was assessed by histopathology of lungs. Cox-2 productions by primary and secondary tumor were measured by real-time RT-PCR and immunoblotting (IB. Proliferation assay based on [3H]-thymidine incorporation and measurement of cytokines and effector molecules by RT-PCR were used to confirm tumor suppression efficacy of abalone visceral extract by modulating cytolytic CD8+ T cells. The cytotoxicity of CD8+ T cell was compared by JAM test. Results Oral administration of abalone visceral extract reduced tumor growth (tumor volume and weight and showed reduced metastasis as confirmed by decreased level of splenomegaly (spleen size and weight and histological analysis of the lung metastasis (gross analysis and histological staining. Reduced expression of Cox-2 (mRNA and protein from primary tumor and metastasized lung was also detected. In addition, treatment of abalone visceral extract increased anti-tumor activities of CD8+ T cells by increasing the proliferation capacity and their cytolytic activity. Conclusions Our results suggest that abalone visceral extract has anti-tumor effects by suppressing tumor growth and lung metastasis through decreasing Cox-2 expression level as well as promoting proliferation and cytolytic function of CD8+ T cells.

  8. From untargeted LC-QTOF analysis to characterisation of opines in abalone adductor muscle: Theory meets practice.

    Science.gov (United States)

    Venter, Leonie; Jansen van Rensburg, Peet J; Loots, Du Toit; Vosloo, Andre; Lindeque, Jeremie Zander

    2017-12-15

    Abalone have a unique ability to use pyruvate, various amino acids and dehydrogenases, to produce opines as means to prevent the accumulation of NADH during anaerobic conditions. In this study, the theoretical masses, formulae and fragment patterns of butylated opines were used to predict which of these compounds could be found in the abalone adductor muscle using untargeted liquid chromatography quadrupole time-of flight-mass spectrometry. These findings were validated using synthesised opine standards. In essence alanopine, lysopine, strombine and tauropine produced in abalone adductor muscle could be characterised using the highest identification confidence levels. Copyright © 2017 Elsevier B.V. All rights reserved.

  9. Microarray analysis of gene expression in disk abalone Haliotis discus discus after bacterial challenge.

    Science.gov (United States)

    De Zoysa, Mahanama; Nikapitiya, Chamilani; Oh, Chulhong; Lee, Youngdeuk; Whang, Ilson; Lee, Jae-Seong; Choi, Cheol Young; Lee, Jehee

    2011-02-01

    In this study, we investigated the gene expression profiling of disk abalone, Haliotis discus discus challenged by a mixture of three pathogenic bacteria Vibrio alginolyticus, Vibrio parahemolyticus, and Listeria monocytogenes using a cDNA microarray. Upon bacteria challenge, 68 (1.6%) and 112 (2.7%) gene transcripts changed their expression levels ≥2 or ≤2 -fold in gills and digestive tract, respectively. There were 46 tissue-specific transcripts that up-regulated specifically in the digestive tract. In contrast, only 13 transcripts showed gill-specific up-regulation. Quantitative real-time PCR was performed to verify microarray data and results revealed that candidate genes namely Krüppell-like factor (KLF), lachesin, muscle lim protein, thioredoxin-2 (TRx-2), nuclear factor interleukin 3 (NFIL-3) and abalone protein 38 were up-regulated. Also, our results further indicated that bacteria challenge may activate the transcription factors or their activators (Krüppell-like factor, inhibitor of NF-κB or Ik-B), inflammatory cytokines (IL-3 regulated protein, allograft inflammatory factor), other cytokines (IFN-44-like protein, SOCS-2), antioxidant enzymes (glutathione-S-transferase, thioredoxin-2 and thioredoxin peroxidase), and apoptosis-related proteins (TNF-α, archeron) in abalone. The identification of immune and stress response genes and their expression profiles in this microarray will permit detailed investigation of the stress and immune responses of abalone genes. Copyright © 2010 Elsevier Ltd. All rights reserved.

  10. Genome Sequence ofArcobactersp. Strain LA11, Isolated from the AbaloneHaliotis discus.

    Science.gov (United States)

    Mizutani, Yukino; Tanaka, Reiji

    2017-03-16

    Arcobacter sp. strain LA11 was isolated from the gut of the abalone Haliotis discus Here, we present the annotation and analysis of the draft genome of this strain, which is involved in nitrogen metabolism. Copyright © 2017 Mizutani and Tanaka.

  11. Más sobre Abalone y algo sobre tuercas: los tetrahexos

    OpenAIRE

    Rupérez, José Antonio; García, Manuel

    2012-01-01

    Comentamos los problemas planteados en anterior artículo sobre jugadas del ABALONE, describiéndolo de nuevo y presentamos el puzle de los Polihexos, fácil de construir y entretenido de resolver; útil para plantear ejercicios de combinatoria y simetrías.

  12. Sperm length, sperm storage and mating system characteristics in bumblebees

    DEFF Research Database (Denmark)

    Baer, Boris; Schmid-Hempel, Paul; Høeg, Jens Thorvald

    2003-01-01

    Multiple insemination induces sperm competition and may select for longer, faster moving sperm in species where sperm is short-lived and egg fertilization takes place almost immediately after ejaculation. Here we report the first detailed analysis of sperm length in social insects with long......-term storage of sperm, using three bumblebee species with different mating systems as models. We show that individual males produce only one size-class of sperm, but that sperm length is highly variable among brothers, among unrelated conspecific males, and among males of different species. Males of Bombus...... hypnorum, a species with multiple-mating queens, have longer sperm than males of B. terrestris and B. lucorum whose queens are single mated. Although the sample size on the species level was too small to perform a phylogenetic analysis, this finding supports the hypothesis that, all other things being...

  13. Subversive practices of sperm donation - globalizing Danish sperm

    DEFF Research Database (Denmark)

    Willum Adrian, Stine

    During the past two decades, Denmark has developed in to an important destination for fertility travellers in need of donor sperm. Furthermore, two of the largest sperm banks in Europe have been established in Denmark, exporting sperm globally. This development has taken place at the same time...

  14. Genome sequence of pacific abalone (Haliotis discus hannai): the first draft genome in family Haliotidae.

    Science.gov (United States)

    Nam, Bo-Hye; Kwak, Woori; Kim, Young-Ok; Kim, Dong-Gyun; Kong, Hee Jeong; Kim, Woo-Jin; Kang, Jeong-Ha; Park, Jung Youn; An, Cheul Min; Moon, Ji-Young; Park, Choul Ji; Yu, Jae Woong; Yoon, Joon; Seo, Minseok; Kim, Kwondo; Kim, Duk Kyung; Lee, SaetByeol; Sung, Samsun; Lee, Chul; Shin, Younhee; Jung, Myunghee; Kang, Byeong-Chul; Shin, Ga-Hee; Ka, Sojeong; Caetano-Anolles, Kelsey; Cho, Seoae; Kim, Heebal

    2017-05-01

    Abalones are large marine snails in the family Haliotidae and the genus Haliotis belonging to the class Gastropoda of the phylum Mollusca. The family Haliotidae contains only one genus, Haliotis, and this single genus is known to contain several species of abalone. With 18 additional subspecies, the most comprehensive treatment of Haliotidae considers 56 species valid [ 1 ]. Abalone is an economically important fishery and aquaculture animal that is considered a highly prized seafood delicacy. The total global supply of abalone has increased 5-fold since the 1970s and farm production increased explosively from 50 mt to 103 464 mt in the past 40 years. Additionally, researchers have recently focused on abalone given their reported tumor suppression effect. However, despite the valuable features of this marine animal, no genomic information is available for the Haliotidae family and related research is still limited. To construct the H . discus hannai genome, a total of 580-G base pairs using Illumina and Pacbio platforms were generated with 322-fold coverage based on the 1.8-Gb estimated genome size of H . discus hannai using flow cytometry. The final genome assembly consisted of 1.86 Gb with 35 450 scaffolds (>2 kb). GC content level was 40.51%, and the N50 length of assembled scaffolds was 211 kb. We identified 29 449 genes using Evidence Modeler based on the gene information from ab initio prediction, protein homology with known genes, and transcriptome evidence of RNA-seq. Here we present the first Haliotidae genome, H . discus hannai , with sequencing data, assembly, and gene annotation information. This will be helpful for resolving the lack of genomic information in the Haliotidae family as well as providing more opportunities for understanding gastropod evolution. © The Authors 2017. Published by Oxford University Press.

  15. Healthy Sperm: Improving Your Fertility

    Science.gov (United States)

    Healthy Lifestyle Getting pregnant Healthy sperm aren't always a given. Understand how lifestyle factors can affect your ... of retrieving and storing sperm before treatment. Adopting healthy lifestyle practices to promote your fertility — and avoiding things ...

  16. Molecular identification of disk abalone (Haliotis discus discus) tetraspanin 33 and CD63: Insights into potent players in the disk abalone host defense system.

    Science.gov (United States)

    Priyathilaka, Thanthrige Thiunuwan; Bathige, S D N K; Herath, H M L P B; Lee, Sukkyoung; Lee, Jehee

    2017-10-01

    Tetraspanins are a superfamily of transmembrane proteins involved in a diverse range of physiological processes including differentiation, adhesion, signal transduction, cell motility, and immune responses. In the present study, two tetraspanins, CD63 and tetraspanin 33 (TSPAN33) from disk abalone (AbCD63 and AbTSPAN33), were identified and characterized at the molecular level. The coding sequences for AbCD63 and AbTSPAN33 encoded polypeptides of 234 and 290 amino acids (aa) with predicted molecular mass of 25.3 and 32.5 kDa, respectively. The deduced AbCD63 and AbTSPAN33 protein sequences were also predicted to have a typical tetraspanin domain architecture, including four transmembrane domains (TM), short N- and C- terminal regions, a short intracellular loop, as well as a large and small extracellular loop. A characteristic CCG motif and cysteine residues, which are highly conserved across CD63 and TSPAN33 proteins of different species, were present in the large extracellular loop of both abalone tetraspanins. Phylogenetic analysis revealed that the AbCD63 and AbTSPAN33 clustered in the invertebrate subclade of tetraspanins, thus exhibiting a close relationship with tetraspanins of other mollusks. The AbCD63 and AbTSPAN33 mRNA transcripts were detected at early embryonic development stages of disk abalone with significantly higher amounts at the trochophore stage, suggesting the involvement of these proteins in embryonic development. Both AbCD63 and AbTSPAN33 were ubiquitously expressed in all the tissues of unchallenged abalones analyzed, with the highest expression levels found in hemocytes. Moreover, significant induction of AbCD63 and AbTSPAN33 mRNA expression was observed in immunologically important tissues, such as hemocytes and gills, upon stimulation with live bacteria (Vibrio parahaemolyticus and Listeria monocytogenes), virus (viral hemorrhagic septicemia virus), and two potent immune stimulators [polyinosinic:polycytidylic acid (poly I:C) and

  17. Sperm whale clicks

    DEFF Research Database (Denmark)

    Møhl, Bertel; Wahlberg, Magnus; Madsen, Peter T.

    2000-01-01

    . A sound generator weighing upward of 10 tons and with a cross-section of 1 m is expected to generate high-intensity, directional sounds. This prediction from the Norris and Harvey theory is not supported by published data for sperm whale clicks ~source levels of 180 dB re 1 mPa and little, if any....... This implicates sonar as a possible function of the clicks. Thus, previously published properties of sperm whale clicks underestimate the capabilities of the sound generator and therefore cannot falsify the Norris and Harvey theory.......In sperm whales ~Physeter catodon L. 1758! the nose is vastly hypertrophied, accounting for about one-third of the length or weight of an adult male. Norris and Harvey @in Animal Orientation and Navigation, NASA SP-262 ~1972!, pp. 397–417# ascribed a sound-generating function to this organ complex...

  18. Subversive practices of sperm donation - globalizing Danish sperm

    DEFF Research Database (Denmark)

    Willum Adrian, Stine

    During the past two decades, Denmark has developed in to an important destination for fertility travellers in need of donor sperm. Furthermore, two of the largest sperm banks in Europe have been established in Denmark, exporting sperm globally. This development has taken place at the same time...... as the use of donated sperm continuously has been debated as an ethical issue, and increasingly been regulated. In this presentation I will discuss how Denmark became a destination for fertility travelling (sperm donation) as a result of various subversive strategies of family making. The article inquires...... into how the bending of boundaries by “inappropriate parents”, fertility travellers, private sperm banks and fertility clinics have been part in negotiating the changes of the legislation in practice, and thus been part of developing a Danish industry of sperm banking. The presentation is based on a multi...

  19. Optimization of lysine metabolism in Corynebacterium glutamicum

    DEFF Research Database (Denmark)

    Rytter, Jakob Vang

    Commercial pig and poultry production use the essential amino acid lysine as a feed additive with the purpose of optimizing the feed utilization. Lysine is produced by a fermentation process involving either Corynebacterium glutamicum or Escherichia coli. The global annual production is around 1......,000,000 tons. The aim of this project is to optimize the yield of lysine in C. glutamicum using metabolic engineering strategies. According to a genome scale model of C. glutamicum, theoretically there is much room for increasing the lysine yield (Kjeldsen and Nielsen 2009). Lysine synthesis requires NADPH...... the PPP, increasing the NADPH synthesis and enabling increased lysine production. Synthetic promoter libraries (SPL) enable fine tuning of the expression of genes. To test the feasibility of SPL in C. glutamicum four constitutive SPLs and one inducible SPL were constructed. The libraries were placed...

  20. Availability of intestinal microbial lysine for whole body lysine homeostasis in human subjects.

    Science.gov (United States)

    Metges, C C; El-Khoury, A E; Henneman, L; Petzke, K J; Grant, I; Bedri, S; Pereira, P P; Ajami, A M; Fuller, M F; Young, V R

    1999-10-01

    We have investigated whether there is a net contribution of lysine synthesized de novo by the gastrointestinal microflora to lysine homeostasis in six adults. On two separate occasions an adequate diet was given for a total of 11 days, and a 24-h (12-h fast, 12-h fed) tracer protocol was performed on the last day, in which lysine turnover, oxidation, and splanchnic uptake were measured on the basis of intravenous and oral administration of L-[1-(13)C]lysine and L-[6,6-(2)H(2)]lysine, respectively. [(15)N(2)]urea or (15)NH(4)Cl was ingested daily over the last 6 days to label microbial protein. In addition, seven ileostomates were studied with (15)NH(4)Cl. [(15)N]lysine enrichment in fecal and ileal microbial protein, as precursor for microbial lysine absorption, and in plasma free lysine was measured by gas chromatography-combustion-isotope ratio mass spectrometry. Differences in plasma [(13)C]- and [(2)H(2)]lysine enrichments during the 12-h fed period were observed between the two (15)N tracer studies, although the reason is unclear, and possibly unrelated to the tracer form per se. In the normal adults, after (15)NH(4)Cl and [(15)N(2)]urea intake, respectively, lysine derived from fecal microbial protein accounted for 5 and 9% of the appearance rate of plasma lysine. With ileal microbial lysine enrichment, the contribution of microbial lysine to plasma lysine appearance was 44%. This amounts to a gross microbial lysine contribution to whole body plasma lysine turnover of between 11 and 130 mg. kg(-1). day(-1), depending on the [(15)N]lysine precursor used. However, insofar as microbial amino acid synthesis is accompanied by microbial breakdown of endogenous amino acids or their oxidation by intestinal tissues, this may not reflect a net increase in lysine absorption. Thus we cannot reliably estimate the quantitative contribution of microbial lysine to host lysine homeostasis with the present paradigm. However, the results confirm the significant presence of

  1. Druggability of methyl-lysine binding sites

    Science.gov (United States)

    Santiago, C.; Nguyen, K.; Schapira, M.

    2011-12-01

    Structural modules that specifically recognize—or read—methylated or acetylated lysine residues on histone peptides are important components of chromatin-mediated signaling and epigenetic regulation of gene expression. Deregulation of epigenetic mechanisms is associated with disease conditions, and antagonists of acetyl-lysine binding bromodomains are efficacious in animal models of cancer and inflammation, but little is known regarding the druggability of methyl-lysine binding modules. We conducted a systematic structural analysis of readers of methyl marks and derived a predictive druggability landscape of methyl-lysine binding modules. We show that these target classes are generally less druggable than bromodomains, but that some proteins stand as notable exceptions.

  2. KARAKTERISASI DAN EVALUASI POPULASI ABALON Haliotis squamata SECARA MOLEKULER, MORFOMETRIK, DAN BIOLOGI

    Directory of Open Access Journals (Sweden)

    Gusti Ngurah Permana

    2017-10-01

    Full Text Available Abalon merupakan salah satu komoditas penting gastropoda laut. Tingginya permintaan abalon ini mengakibatkan menipisnya stok di alam. Oleh karena itu, upaya keberhasilan budidaya abalon perlu didukung oleh jenis unggul. Indikasi awal suatu jenis unggul dapat dilakukan dengan menganalisis potensi genetik yang dimiliki. Penelitian ini dilakukan dengan analisis gen 16S rRNA, karakter morfolologi, dan biologi dianalisis secara deskriptif dengan metode kajian pustaka. Hasil yang diperoleh menunjukkan keragaman inter populasi Haliotis squamata mendeteksi adanya tujuh haplotipe yang terbagi dalam dua kelompok. Penyertaan H. diversicolor sebagai outgroup dalam pengujian memperlihatkan bahwa populasi H. squamata dari Pulau Bali dan beberapa lokasi di Pulau Jawa berada dalam satu kelompok yang terpisah dengan outgroup. Hasil ini kongruen dengan analisis morfometrik terdapat perkembangan pertumbuhan cangkang yang asimetri pada populasi Banten. Pertumbuhan asimetri merupakan indikasi spesifik untuk populasi Banten atau merupakan gejala abnormalitas yang dapat diakibatkan oleh faktor penurunan kualitas genetik atau lingkungan. Karakter biologi terlihat proporsi daging dan gonad berbeda pada populasi Banten dengan indikasi adanya pertumbuhan asimetri. Rasio gonad dan daging populasi Banyuwangi berbeda nyata (P<0,05 dengan populasi lainnya. Abalone is arguably one of the highly valued and sought-after marine gastropods. However, the over-exploitation of this species has exhausted its wild stock. To overcome this challenge, the culture technique and management of this species must be established and continually improved. One of the ways is through producing superior broodstocks. An initial assessment of a genetically superior broodstock can be done using the potential genetic analysis. This recent research employed the analysis to study the species’ 16S rRNA gene. To complement the study, the morphometric and biological characteristics of the species were

  3. Hemoglobin Labeled by Radioactive Lysine

    Science.gov (United States)

    Bale, W. F.; Yuile, C. L.; DeLaVergne, L.; Miller, L. L.; Whipple, G. H.

    1949-12-08

    This paper reports on the utilization of tagged epsilon carbon of DL-lysine by a dog both anemic and hypoproteinemic due to repeated bleeding plus a diet low in protein. The experiment extended over period of 234 days, a time sufficient to indicate an erythrocyte life span of at least 115 days based upon the rate of replacement of labeled red cell proteins. The proteins of broken down red cells seem not to be used with any great preference for the synthesis of new hemoglobin.

  4. Sperm head's birefringence: a new criterion for sperm selection.

    Science.gov (United States)

    Gianaroli, Luca; Magli, M Cristina; Collodel, Giulia; Moretti, Elena; Ferraretti, Anna P; Baccetti, Baccio

    2008-07-01

    To investigate the characteristics of birefringence in human sperm heads and apply polarization microscopy for sperm selection at intracytoplasmic sperm injection (ICSI). Prospective randomized study. Reproductive Medicine Unit, Società Italiana Studi Medicina della Riproduzione, Bologna, Italy. A total of 112 male patients had birefringent sperm selected for ICSI (study group). The clinical outcome was compared with that obtained in 119 couples who underwent a conventional ICSI cycle (control group). The proportion of birefringent spermatozoa was evaluated before and after treatment in relation to the sperm sample quality. Embryo development and clinical outcome in the study group were compared with those in the controls. Proportion of birefringent sperm heads, rates of fertilization, cleavage, pregnancy, implantation, and ongoing implantation. The proportion of birefringent spermatozoa was significantly higher in normospermic samples when compared with oligoasthenoteratospermic samples with no progressive motility and testicular sperm extraction samples. Although fertilization and cleavage rates did not differ between the study and control groups, in the most severe male factor condition (oligoasthenoteratospermic with no progressive motility and testicular sperm extraction), the rates of clinical pregnancy, ongoing pregnancy, and implantation were significantly higher in the study group versus the controls. The analysis of birefringence in the sperm head could represent both a diagnostic tool and a novel method for sperm selection.

  5. Gene expression profiling in respond to TBT exposure in small abalone Haliotis diversicolor.

    Science.gov (United States)

    Jia, Xiwei; Zou, Zhihua; Wang, Guodong; Wang, Shuhong; Wang, Yilei; Zhang, Ziping

    2011-10-01

    In this study, we investigated the gene expression profiling of small abalone, Haliotis diversicolor by tributyltin (TBT) exposure using a cDNA microarray containing 2473 unique transcripts. Totally, 107 up-regulated genes and 41 down-regulated genes were found. For further investigation of candidate genes from microarray data and EST analysis, quantitative real-time PCR was performed at 6 h, 24 h, 48 h, 96 h and 192 h TBT exposure. 26 genes were found to be significantly differentially expressed in different time course, 3 of them were unknown. Some gene homologues like cellulose, endo-beta-1,4-glucanase, ferritin subunit 1 and thiolester containing protein II CG7052-PB might be the good biomarker candidate for TBT monitor. The identification of stress response genes and their expression profiles will permit detailed investigation of the defense responses of small abalone genes. Published by Elsevier Ltd.

  6. Immune priming and portal of entry effectors improve response to vibrio infection in a resistant population of the European abalone.

    Science.gov (United States)

    Dubief, Bruno; Nunes, Flavia L D; Basuyaux, Olivier; Paillard, Christine

    2017-01-01

    Since 1997, populations of the European abalone Haliotis tuberculata suffer mass mortalities attributed to the bacterium Vibrio harveyi. These mortalities occur at the spawning season, when the abalone immune system is depressed, and when temperatures exceed 17 °C, leading to favorable conditions for V. harveyi proliferation. In order to identify mechanisms of disease resistance, experimental successive infections were carried out on two geographically distinct Brittany populations: one that has suffered recurrent mortalities (Saint-Malo) and one that has not been impacted by the disease (Molène). Furthermore, abalone surviving these two successive bacterial challenges and uninfected abalone were used for several post-infection analyses. The Saint-Malo population was found to be resistant to V. harveyi infection, with a survival rate of 95% compared to 51% for Molène. While in vitro quantification of phagocytosis by flow cytometry showed strong inhibition following the first infection, no inhibition of phagocytosis was observed following the second infection for Saint-Malo, suggesting an immune priming effect. Moreover, assays of phagocytosis of GFP-labelled V. harveyi performed two months post-infection show an inhibition of phagocytosis by extracellular products of V. harveyi for uninfected abalone, while no effect was observed for previously infected abalone from Saint-Malo, suggesting that the effects of immune priming may last upwards of two months. Detection of V. harveyi by qPCR showed that a significantly greater number of abalone from the susceptible population were positive for V. harveyi in the gills, indicating that portal of entry effectors may play a role in resistance to the disease. Collectively, these results suggest a potential synergistic effect of gills and hemolymph in the resistance of H. tuberculata against V. harveyi with an important involvement of the gills, the portal of entry of the bacteria. Copyright © 2016 Elsevier Ltd

  7. PENILAIAN PENGARUH PENAMBAHAN LYSINE PADA NASI

    Directory of Open Access Journals (Sweden)

    Ignatius Tarwotjo

    2012-11-01

    Full Text Available Pengaruh penambahan lysine pada mutu protein nasi dilakukan pada tikus putih dengan mengukur Protein Efficiency Ratio. Nasi dan Nasi dengan sayur beserta laukpauk, seperti dikonsumsi oleh kebanyakan keluarga di Indonesia, yang berasnya lebih dulu ditambahi butiran premix berisi lysine, thiamine dan riboflavin ternaya menghasilkan Protein Efficiency Ratio lebih tinggi dari pada yang tidak ditambahi.

  8. Microbial production of lysine from sustainable feedstock

    DEFF Research Database (Denmark)

    Wang, Zhihao; Grishkova, Maria; Solem, Christian

    2014-01-01

    Lysine is produced in a fermentation process using Corynebacterium glutamicum. And even though production strains have been improved for decades, there is still room for further optimization.......Lysine is produced in a fermentation process using Corynebacterium glutamicum. And even though production strains have been improved for decades, there is still room for further optimization....

  9. Availability of phosphorus from selected inorganic phosphate to juvenile abalone, Haliotis discus hannai ino.

    Science.gov (United States)

    Tan, Bei-Ping; Mai, Kang-Sen; Xu, Wei

    2002-06-01

    The availability of phosphorus to juvenile abalone, Haliotis discus hannai from primary, secondary and tertiary calcium phosphate, primary sodium or potassium phosphate separately or in combination was determined in a 120-day feeding trial. Seven semi-purified diets were formulated to contain relatively constant dietary phosphorus, ranging from 0.78% 0.82%. The concentrations of dietary phosphorus were presumed to be slightly below the requirement for abalone. A brown alga, Laminaria japonica, was used as a control diet. Abalone juveniles of silimar size (average weight 1.18 g; average shell length 18.74 mm) were distributed in a single-pass, flow-through system using a completely randomized design with eight treatments and three replicates each treatment. The survival ranged from 71.1% to 81.1%, and was not significantly (P>0.05) related to dietary treatment. However, the weight gain rate (41.72% 65.02%), daily increment in shell length (36.87 55.07 μm) and muscle RNA-DNA ratio (3.44 4.69) were significantly (Pdietary treatment. Soft body alkaline phosphatase activity (10.9 19.8 U/g wet tissue) and carcass levels of lipid (7.71% 9.33%) and protein (46.68% 49.35%) were significantly (PDietary treatment had significant effect (Pdietary treatments. The results indicated that among these inorganic compounds, primary calcium, potassium and sodium phosphates separately or in combination could be utilized effectively by juvenile abalone as dietary phosphorus sources (availability ranging from 87% 97%). However, secondary and tertiary calcium phosphates were found to be low in availability, the values being 45% and 77%, repectively.

  10. Abalone, Haliotis mariae (Wood, 1828, Hatchery and Seed Production Trials in Oman

    Directory of Open Access Journals (Sweden)

    Khalfan M. Al-Rashdi

    2008-01-01

    Full Text Available Hatchery and seed production trials for the Omani endemic abalone Haliotis mariae were carried out at the land-based Mirbat Abalone Seed Production Station in Oman between 1999 and 2000. The methods developed for broodstock conditioning, induction of spawning and fertilization, larval settlement, and the handling of small juveniles are shown. Abalone collected in the post-monsoon period and held for 2 months matured faster than those collected before the monsoon and held for 6 months. Spawning induction of males and females had 63% and 11% success rates respectively, and the morphology of early larval stages is shown. Survival rates of veliger larvae introduced to settlement plates ranged from 35.9% to 73.7%, but the survival of post-larvae was low at 0.1% to 3.6%. The high mortality rate was attributed to invasions of filamentous green- and coralline algae on settlement plates and occurrence of low quantity of diatoms as food. Juveniles reacted best to 2% ethanol as anaesthetic, dropping off culture plates within 4 min and recovering within 17 min. Cultured abalone reached an average shell length of 52.9 mm over 13 months, which translates to an increment of 4.1 mm.mon-1. The overall conclusion of these preliminary research trials confirms  that H. mariae can be cultured successfully in Oman. Further studies on the standardization of the techniques would help in stock enhancement programmes and commercial farming.

  11. Two ferritin subunits from disk abalone (Haliotis discus discus): cloning, characterization and expression analysis.

    Science.gov (United States)

    De Zoysa, Mahanama; Lee, Jehee

    2007-09-01

    Ferritin plays a key role in cellular iron metabolism, which includes iron storage and detoxification. From disk abalone, Haliotis discus discus, the cDNA that encodes the two ferritin subunits abalone ferritin subunit 1 (Abf1) and abalone ferritin subunit 2 (Abf2) were cloned. The complete cDNA coding sequences for Abf1 and Abf2 contained 621 and 549 bp, encoding for 207 and 183 amino acid residues, respectively. The H. discus discus Abf2 subunit contained a highly conserved motif for the ferroxidase center, which consists of seven residues of a typical vertebrate heavy-chain ferritin with a typical stem-loop structure. Abf2 mRNA contains a 27 bp iron-responsive element (IRE) in the 5'UTR position. This IRE exhibited 96% similarity with pearl and Pacific oyster and 67% similarity with human H type IREs. However, the Abf1 subunit had neither ferroxidase center residues nor the IRE motif sequence; instead, it contained iron-binding region signature 2 (IBRS) residues. Recombinant Abf1 and Abf2 proteins were purified and the respective sizes were about 24 and 21 kDa. Abf1 and Abf2 exhibited iron-chelating activity 44.2% and 22.0%, respectively, at protein concentration of 6 microg/ml. Analysis of tissue-specific expression by RT-PCR revealed that Abf1 and Abf2 ferritin mRNAs were expressed in various abalone tissues, such as gill, mantle, gonad, foot and digestive tract in a wide distribution profile, but Abf2 expression was more prominent than Abf1.

  12. Evolution of large body size in abalones (Haliotis): Patterns and implications

    Science.gov (United States)

    Estes, J.A.; Lindberg, D.R.; Wray, C.

    2005-01-01

    Kelps and other fleshy macroalgae - dominant reef-inhabiting organisms in cool - seasmay have radiated extensively following late Cenozoic polar cooling, thus triggering a chain of evolutionary change in the trophic ecology of nearshore temperate ecosystems. We explore this hypothesis through an analysis of body size in the abalones (Gastropoda; Haliotidae), a widely distributed group in modern oceans that displays a broad range of body sizes and contains fossil representatives from the late Cretaceous (60-75 Ma). Geographic analysis of maximum shell length in living abalones showed that small-bodied species, while most common in the Tropics, have a cosmopolitan distribution, whereas large-bodied species occur exclusively in cold-water ecosystems dominated by kelps and other macroalgae. The phylogeography of body size evolution in extant abalones was assessed by constructing a molecular phylogeny in a mix of large and small species obtained from different regions of the world. This analysis demonstrates that small body size is the plesiomorphic state and largeness has likely arisen at least twice. Finally, we compiled data on shell length from the fossil record to determine how (slowly or suddenly) and when large body size arose in the abalones. These data indicate that large body size appears suddenly at the Miocene/Pliocene boundary. Our findings support the view that fleshy-algal dominated ecosystems radiated rapidly in the coastal oceans with the onset of the most recent glacial age. We conclude with a discussion of the broader implications of this change. ?? 2005 The Paleontological Society. All rights reserved.

  13. Comparative evidence for the evolution of sperm swimming speed by sperm competition and female sperm storage duration in passerine birds.

    Science.gov (United States)

    Kleven, Oddmund; Fossøy, Frode; Laskemoen, Terje; Robertson, Raleigh J; Rudolfsen, Geir; Lifjeld, Jan T

    2009-09-01

    Sperm swimming speed is an important determinant of male fertility and sperm competitiveness. Despite its fundamental biological importance, the underlying evolutionary processes affecting this male reproductive trait are poorly understood. Using a comparative approach in a phylogenetic framework, we tested the predictions that sperm swim faster with (1) increased risk of sperm competition, (2) shorter duration of female sperm storage, and (3) increased sperm length. We recorded sperm swimming speed in 42 North American and European free-living passerine bird species, representing 35 genera and 16 families. We found that sperm swimming speed was positively related to the frequency of extrapair paternity (a proxy for the risk of sperm competition) and negatively associated with clutch size (a proxy for the duration of female sperm storage). Sperm swimming speed was unrelated to sperm length, although sperm length also increased with the frequency of extrapair paternity. These results suggest that sperm swimming speed and sperm length are not closely associated traits and evolve independently in response to sperm competition in passerine birds. Our findings emphasize the significance of both sperm competition and female sperm storage duration as evolutionary forces driving sperm swimming speed.

  14. Adipic acid increases plasma lysine but does not improve the efficiency of lysine utilization in swine.

    Science.gov (United States)

    van Kempen, T A; van Heugten, E; Trottier, N L

    2001-09-01

    Adipic acid, upon catabolism, results in intermediates that bear a structural similarity to lysine degradation products. The objectives of this research were to determine whether adipic acid affects lysine concentrations in plasma and to evaluate whether adipic acid improves the efficiency of lysine utilization in pigs. In Exp. 1, nursery pigs (n = 14) were fed (for a period of 7 d) either a standard nursery diet or the same diet supplemented with 1% adipic acid to assess effects on plasma amino acid concentrations (plasma collected on d 7). In Exp. 2, nursery pigs (n = 56) were fed (for a period of 15 d) either a control diet or the same diet but deficient in either lysine, threonine, or tryptophan with or without supplemental adipic acid to assess the effects of adipic acid on the efficiency of amino acid utilization. The results from Exp. 1 showed that adipic acid increased plasma lysine (by 18%) but not alpha-amino adipic acid, an intermediate in lysine degradation. Experiment 2 demonstrated that adipic acid did not increase the efficiency of utilization of lysine, threonine, or tryptophan. The lack of effects on alpha-amino adipic acid in Exp. 1 and the lack of a positive effect on the efficiency of utilization of lysine, threonine, and tryptophan suggest that adipic acid does not inhibit the mitochondrial uptake of lysine and(or) its degradation in the mitochondrion. It is concluded that feeding adipic acid increases plasma lysine but does not improve the efficiency of lysine utilization.

  15. Matrix metalloproteinases (MMPs) inhibitory effects of an octameric oligopeptide isolated from abalone Haliotis discus hannai.

    Science.gov (United States)

    Nguyen, Van-Tinh; Qian, Zhong-Ji; Ryu, Bomi; Kim, Kil-Nam; Kim, Daekyung; Kim, Young-Mog; Jeon, You-Jin; Park, Won Sun; Choi, Il-Whan; Kim, Geun Hyung; Je, Jae-Young; Jung, Won-Kyo

    2013-11-01

    Abalone (Haliotis discus hannai) is a marine gastropod, and an important fishery and food industrial resource that is massively maricultured in Asia, Africa, Australia and America. However, its health benefits have rarely been studied for nutraceutical and pharmaceutical application. In this study, the purified abalone oligopeptide (AOP) with anti-matrix metalloproteinases (anti-MMPs) effects was isolated from the digests of abalone intestine using recycle HPLC with a JAI W253 column and an OHpak SB-803 HQ column. The AOP was identified as Ala-Glu-Leu-Pro-Ser-Leu-Pro-Gly (MW=782.4 Da) with a de novo peptide sequencing technique using a tandem mass spectrometer. The AOP exhibited a specific inhibitory effect against MMP-2/-9 activity and attenuated protein expression of p50 and p65 in the human fibrosarcoma (HT1080) cells, dose-dependently. The results presented illustrate that the AOP could inhibit MMP-2/-9 expression in HT1080 cells via the nuclear factor-kappaB (NF-κB)-mediated pathway. This data suggest that the AOP from H. discus hannai intestine may possess therapeutic and preventive potential for the treatment of MMPs-related disorders such as angiogenesis and cardiovascular diseases. Copyright © 2013 Elsevier Ltd. All rights reserved.

  16. Sperm cell proteomics.

    Science.gov (United States)

    Oliva, Rafael; de Mateo, Sara; Estanyol, Josep Maria

    2009-02-01

    The spermatozoon is an accessible cell which can be easily purified and therefore it is particularly well suited for proteomic analysis. It is also an extremely differentiated cell with very marked genetic, cellular, functional and chromatin changes as compared to other cells, and has profound implications for fertility, embryo development and heredity. The recent developments in MS have boosted the potential for identification and study of the sperm proteins. Catalogues of hundreds to thousands of spermatozoan proteins in human and in model species are becoming available setting up the basis for subsequent research, diagnostic applications and the development of specific treatments. The present article reviews the available scientific publications dealing with the composition and function of the sperm cell using an MS proteomic approach.

  17. Cryopreservation of Fish Sperm

    Science.gov (United States)

    Kurokura, Hisashi

    Present status of research activities in cryopreservation of fish gamete in aquaculture field was introduced. More than 59 fish species have been reported in the research histories and nearly half of them were studied during recent 10 years. This means that the research activities are increasing, though commercial profit have not obtained yet. Fish species of which sperm can successfully cryopreserved is still limited comparing to numerous species in telost. One of the major obstacle for improvement of the technique is existence of wide specie specific variance in the freezing tolerance of fish sperm. The varianc can possibly be explaind thorugh the informations obtained by the studies in comparative spermatology, which is recently activated field in fish biology.

  18. Morphology, Structure of Dimorphic Sperm, and Reproduction in the Hermaphroditic Commensal Bivalve Pseudopythina tsurumaru (Galeommatoidea: Kellidae)

    DEFF Research Database (Denmark)

    Lützen, Jørgen; Jespersen, Åse; Takahashi, Tohru

    2004-01-01

    Galeommatoide, commensal bivalve, reproduction, dimorphic sperm, sperm ultrastructure, spermatozeugma......Galeommatoide, commensal bivalve, reproduction, dimorphic sperm, sperm ultrastructure, spermatozeugma...

  19. Relative testis size and sperm morphometry across mammals: no evidence for an association between sperm competition and sperm length.

    OpenAIRE

    Gage, Matthew J G; Freckleton, Robert P

    2003-01-01

    Understanding why there is extensive variation in sperm form and function across taxa has been a challenge because sperm are specialized cells operating at a microscopic level in a complex environment. This comparative study collates published data to determine whether the evolution of sperm morphometry (sperm total length and separate component dimensions) is associated with sperm competition (when different males' sperm mix and compete for a female's ova) across 83 mammalian species. We use...

  20. Sperm function test

    Directory of Open Access Journals (Sweden)

    Pankaj Talwar

    2015-01-01

    Full Text Available With absolute normal semen analysis parameters it may not be necessary to shift to specialized tests early but in cases with borderline parameters or with history of fertilization failure in past it becomes necessary to do a battery of tests to evaluate different parameters of spermatozoa. Various sperm function tests are proposed and endorsed by different researchers in addition to the routine evaluation of fertility. These tests detect function of a certain part of spermatozoon and give insight on the events in fertilization of the oocyte. The sperms need to get nutrition from the seminal plasma in the form of fructose and citrate (this can be assessed by fructose qualitative and quantitative estimation, citrate estimation. They should be protected from the bad effects of pus cells and reactive oxygen species (ROS (leukocyte detection test, ROS estimation. Their number should be in sufficient in terms of (count, structure normal to be able to fertilize eggs (semen morphology. Sperms should have intact and functioning membrane to survive harsh environment of vagina and uterine fluids (vitality and hypo-osmotic swelling test, should have good mitochondrial function to be able to provide energy (mitochondrial activity index test. They should also have satisfactory acrosome function to be able to burrow a hole in zona pellucida (acrosome intactness test, zona penetration test. Finally, they should have properly packed DNA in the nucleus to be able to transfer the male genes (nuclear chromatic decondensation test to the oocyte during fertilization.

  1. Cytometry of mammalian sperm

    International Nuclear Information System (INIS)

    Gledhill, B.L.

    1983-01-01

    Male germ cells respond dramatically to a variety of insults and are important reproductive dosimeters. Semen analyses are very useful in studies on the effects of drugs, chemicals, and environmental hazards on testicular function, male fertility and heritable germinal mutations. The accessibility of male cells makes them well suited for analytical cytology. We might automate the process of determining sperm morphology but should not do so solely for increased speed. Rather, richer tangible benefits will derive from cytometric evaluation through increased sensitivity, reduced subjectivity, standardization between investigators and laboratories, enhanced archival systems, and the benefits of easily exchanged standardized data. Inroads on the standardization of assays for motility and functional integrity are being made. Flow cytometric analysis of total DNA content of individual sperm is an insensitive means to detect exposure to reproductive toxins because of the small size and low frequency of the DNA content errors. Flow cytometry can be applied to determine the proportions of X- and Y-sperm in semen samples

  2. Cytometry of mammalian sperm

    Energy Technology Data Exchange (ETDEWEB)

    Gledhill, B.L.

    1983-10-11

    Male germ cells respond dramatically to a variety of insults and are important reproductive dosimeters. Semen analyses are very useful in studies on the effects of drugs, chemicals, and environmental hazards on testicular function, male fertility and heritable germinal mutations. The accessibility of male cells makes them well suited for analytical cytology. We might automate the process of determining sperm morphology but should not do so solely for increased speed. Rather, richer tangible benefits will derive from cytometric evaluation through increased sensitivity, reduced subjectivity, standardization between investigators and laboratories, enhanced archival systems, and the benefits of easily exchanged standardized data. Inroads on the standardization of assays for motility and functional integrity are being made. Flow cytometric analysis of total DNA content of individual sperm is an insensitive means to detect exposure to reproductive toxins because of the small size and low frequency of the DNA content errors. Flow cytometry can be applied to determine the proportions of X- and Y-sperm in semen samples.

  3. Black, green, and red abalones. Species profiles: life histories and environmental requirements of coastal fishes and invertebrates (Pacific Southwest. )

    Energy Technology Data Exchange (ETDEWEB)

    Ault, J.S.

    1985-03-01

    Black, green, and red abalones (Haliotis cracherodii, H. fulgens, and H. rufescens, respectivley) are of commercial and ecological importance and are distributed widely along the California coast. The abalones are morphologically similar; species are distinguished by particular shell sculpture, color, and body characteristics. Their latitudinal and bathymetric distribution is stratified and most closely related to temperature. Small juveniles eat mainly microflora; adults eat primarily drift macro-algae, preferring specific brown or red algae, when available. Spawning occurs during summer; gonad ripening depends on food quality and quantity and water temperature. Larvae are lecithotrophic and remain planktonic for periods of 5 to 14 days after hatching; settling is substrate specific. Postlarvae and adults require hard substrate for attachment. Juveniles are cryptic, adults usually more exposed. Growth rates are similar, although maximum size varies with species. Increases in shell length and body weight correlate positively with food abundance and temperature. Below depths of 6 m, sea urchins are major competitors for food and space. Predation by invertebrates is low. Decreased abalone production from central California is associated with range expansion and increased predation by sea otters, the major source of abalone mortality. General declines in California landings are due to mortality from improper picking and replacement, habitat degradation, and perhaps overfishing. Commercial and sport diving efforts have increased sharply, whereas annual landings of abalones declined from 1965 to 1982.

  4. Alternative Splicing Profile and Sex-Preferential Gene Expression in the Female and Male Pacific Abalone Haliotis discus hannai.

    Science.gov (United States)

    Kim, Mi Ae; Rhee, Jae-Sung; Kim, Tae Ha; Lee, Jung Sick; Choi, Ah-Young; Choi, Beom-Soon; Choi, Ik-Young; Sohn, Young Chang

    2017-03-09

    In order to characterize the female or male transcriptome of the Pacific abalone and further increase genomic resources, we sequenced the mRNA of full-length complementary DNA (cDNA) libraries derived from pooled tissues of female and male Haliotis discus hannai by employing the Iso-Seq protocol of the PacBio RSII platform. We successfully assembled whole full-length cDNA sequences and constructed a transcriptome database that included isoform information. After clustering, a total of 15,110 and 12,145 genes that coded for proteins were identified in female and male abalones, respectively. A total of 13,057 putative orthologs were retained from each transcriptome in abalones. Overall Gene Ontology terms and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways analyzed in each database showed a similar composition between sexes. In addition, a total of 519 and 391 isoforms were genome-widely identified with at least two isoforms from female and male transcriptome databases. We found that the number of isoforms and their alternatively spliced patterns are variable and sex-dependent. This information represents the first significant contribution to sex-preferential genomic resources of the Pacific abalone. The availability of whole female and male transcriptome database and their isoform information will be useful to improve our understanding of molecular responses and also for the analysis of population dynamics in the Pacific abalone.

  5. Cryopreservation of epididymal stallion sperm.

    Science.gov (United States)

    Olaciregui, M; Gil, L; Montón, A; Luño, V; Jerez, R A; Martí, J I

    2014-02-01

    Any event that makes semen collection or mating impossible, such as death, castration, or injury, may terminate a stallion's breeding career. Fortunately, stallion sperm which are capable of fertilization can be harvested from the epididymis, and frozen for future use. However, the fertility of frozen-thawed epididymal sperm has been found to be lower than that of ejaculated sperm. Therefore, this study aimed to optimize the fertility of frozen epididymal stallion sperm by investigating the effects of different cryoprotectants and freezing protocols on sperm quality. Dimethylformamide was tested alone or combination with pasteurized egg yolk as substitute of fresh egg yolk. In addition, the effect of the pre-freeze stabilization on sperm quality was analyzed. Heterospermic samples obtained from stallion epididymis were collected and cryopreserved in lactose-egg-yolk extender or in the same extender with varying content of cryoprotectant and content of egg yolk, stabilized and no-stabilized. Sperm motility, viability, hypoosmotic swelling test (HOST) and acrosome integrity were evaluated post-thawing. No improvement was observed on the replacement of fresh yolk by pasteurized egg yolk, whereas the results suggest that dimethylformamide is a cryoprotectant suitable for cryopreservation of equine epididymal semen, even better than glycerol. In addition, we found that the stabilization before freezing on epididymal stallion sperm, can improve sperm quality parameters. Copyright © 2014 Elsevier Inc. All rights reserved.

  6. Daily Sperm Production, Gonadal and Extra-Gonadal Sperm ...

    African Journals Online (AJOL)

    SH

    significantly (P<0.05) higher in rabbits fed prebiotic and symbiotic diets than those fed diet 3 and the control. The daily sperm production and sperm ... The amount of good quality live spermatozoa produced by the testis and the ... production and reproductive performance of rabbits. Effects of nutrition on reproduction.

  7. Unraveling the Sperm Bauplan: Relationships Between Sperm Head Morphology and Sperm Function in Rodents.

    Science.gov (United States)

    Varea-Sánchez, María; Tourmente, Maximiliano; Bastir, Markus; Roldan, Eduardo R S

    2016-07-01

    Rodents have spermatozoa with features not seen in other species. Sperm heads in many rodent species bear one or more apical extensions known as "hooks." The process by which hooks have evolved, together with their adaptive significance, are still controversial issues. In order to improve our understanding of the biological meaning of these sperm head adaptations, we analyzed hook curvature angles, hook length, and overall hook shape in muroid rodents by using geometric morphometrics. We also searched for relationships between hook design and measurements of intermale competition to assess whether postcopulatory sexual selection was an important selective force driving changes in this sperm structure. Finally, we sought possible links between aspects of sperm hook design and sperm velocity as a measure of sperm performance. Results showed that one hook curvature angle is under strong selective pressure. Similarly, hook length appears to be strongly selected by sexual selection, with this selective force also exhibiting a stabilizing role reducing intermale variation in this trait. The adaptive significance of changes in hook structure was supported by the finding that there are strong and significant covariations between hook dimensions and shape and between hook design and sperm swimming velocity. Overall, this study strongly suggests that postcopulatory sexual selection has an important effect on the design of the sperm head that, in turn, is important for enhancing sperm velocity, a function crucial to reaching the vicinity of the female gamete and winning fertilizations under competitive situations. © 2016 by the Society for the Study of Reproduction, Inc.

  8. Factors influencing boar sperm cryosurvival.

    Science.gov (United States)

    Roca, J; Hernández, M; Carvajal, G; Vázquez, J M; Martínez, E A

    2006-10-01

    Optimal sperm cryopreservation is a prerequisite for the sustainable commercial application of frozen-thawed boar semen for AI. Three experiments were performed to identify factors influencing variability of postthaw sperm survival among 464 boar ejaculates. Sperm-rich ejaculate fractions were cryopre-served using a standard freezing-thawing procedure for 0.5-mL plastic straws and computer-controlled freezing equipment. Postthaw sperm motility (assessed with a computer-assisted semen analysis system) and viability (simultaneously probed by flow cytometry analysis after triple-fluorescent stain), evaluated 30 and 150 min postthaw, were used to estimate the success of cryopreservation. In the first experiment, 168 unselected ejaculates (1 ejaculate/boar), from boars of 6 breeds with a wide age range (8 to 48 mo), were cryopreserved over a 12-mo period to evaluate the predictive value of boar (breed and age), semen collection, transport variables (season of ejaculate collection, interval between collections, and ejaculate temperature exposure), initial semen traits, and sperm quality before freezing on sperm survival after freezing-thawing. In Exp. 2, 4 ejaculates from each of 29 boars, preselected according to their initial semen traits and sperm quality before freezing, were collected and frozen over a 6-mo period to evaluate the influence of interboar and intraboar ejaculate variability in the survival of sperm after cryopreservation. In Exp. 3, 12 ejaculates preselected as for Exp. 2, from each of 15 boars with known good sperm cryosurvival, were collected and frozen over a 12-mo period to estimate the sustainability of sperm cryosurvival between ejaculates over time. Boar and semen collection and transport variables were not predictive of sperm cryosurvival among ejaculates. Initial semen traits and sperm quality variables observed before freezing explained 23.2 and 10.9%, respectively, of the variation in postthaw sperm motility and viability. However, more that

  9. Hyaluronic acid binding ability of human sperm reflects cellular maturity and fertilizing potential: selection of sperm for intracytoplasmic sperm injection.

    Science.gov (United States)

    Huszar, Gabor; Ozkavukcu, Sinan; Jakab, Attila; Celik-Ozenci, Ciler; Sati, G Leyla; Cayli, Sevil

    2006-06-01

    The current concepts of sperm biochemical markers and the central role of the HspA2 chaperone protein, a measure of sperm cellular maturity and fertilizing potential, are reviewed. Because HspA2 is a component of the synaptonemal complex, low HspA2 levels and increased frequency of chromosomal aneuploidies are related in diminished maturity sperm. We also suggest a relationship between HspA2 expression in elongating spermatids and events of late spermiogenesis, such as cytoplasmic extrusion and plasma membrane remodeling that aid the formation of the zona pellucida binding and hyaluronic acid binding sites. The presence of hyaluronic acid receptor on the plasma membrane of mature sperm, coupled with hyaluronic acid coated glass or plastic surfaces, facilitates testing of sperm function and selection of single mature sperm for intracytoplasmic sperm injection. The frequencies of sperm with chromosomal disomy are reduced approximately fourfold to fivefold in hyaluronic acid selected sperm compared with semen sperm, comparable to the increase in such abnormalities in intracytoplasmic sperm injection offspring. Hyaluronic acid binding also excludes immature sperm with cytoplasmic extrusion, persistent histones, and DNA chain breaks. Hyaluronic acid mediated sperm selection is a novel technique that is comparable to sperm zona pellucida binding. Hyaluronic acid selected sperm will also alleviate the risks related to intracytoplasmic sperm injection fertilization with sperm of diminished maturity that currently cause worldwide concern.

  10. Induced High Lysine Mutants in Barley

    DEFF Research Database (Denmark)

    Doll, Hans; Køie, B.; Eggum, B. O.

    1974-01-01

    variety. Comparisons of six high lysine mutants with the parent variety showed that grain yield and seed size of the mutants are reduced between 10 and 30 per cent. However, the most promising mutant had the lowest reduction in grain yield, and the absolute lysine yield of this mutant was some 30 per cent...... above that of the parent variety. Feeding tests with rats revealed substantial increases in the biological value of the high lysine mutant protein. Also the net protein utilization was improved but less so because of a somewhat reduced digestibility of the mutant protein....

  11. Methods of sperm vitality assessment.

    Science.gov (United States)

    Moskovtsev, Sergey I; Librach, Clifford L

    2013-01-01

    Sperm vitality is a reflection of the proportion of live, membrane-intact spermatozoa determined by either dye exclusion or osmoregulatory capacity under hypo-osmotic conditions. In this chapter we address the two most common methods of sperm vitality assessment: eosin-nigrosin staining and the hypo-osmotic swelling test, both utilized in clinical Andrology laboratories.

  12. Social imaginaries, sperm and whiteness

    DEFF Research Database (Denmark)

    Andreassen, Rikke

    2017-01-01

    This article analyses narratives about so-called Viking babies and Viking sperm. Over the last few years an increasing number of British single women and lesbian couples have been creating families by becoming pregnant with Danish donor sperm, termed ‘Viking sperm’. Through analyses of British...

  13. Effect of Abalone Hydrolysates Encapsulated by Double Emulsion on the Physicochemical and Sensorial Properties of Fresh Cheese.

    Science.gov (United States)

    Choi, HeeJeong; Kim, Soo-Jin; Lee, Sang-Yoon; Choi, Mi-Jung

    2017-01-01

    The intake of dietary salt through food now exceeds current nutritional recommendations and is thought to have negative effects on human health, such as the increasing prevalence of hypertension. This study was performed to investigate whether W 1 /O/W 2 double emulsions can be used to enhance the saltiness of cheese without increasing the salt content (W 1 is distilled water or 1% abalone hydrolysate, and W 2 is 1% NaCl or 1% abalone hydrolysate + 1% NaCl solution). We also investigated the effect of adding abalone hydrolysate to the double emulsion as a saltiness enhancer. The cheeses were physico-chemically evaluated to determine curd yield, pH value, moisture content, color, texture, salt release rate, and sensory properties. No significant differences were observed in curd yield, pH value, moisture content, lightness, or redness between the cheeses made with and without the double emulsion. However, in the evaluation of salt release rate, fresh cheese made with double emulsion (W 1 = distilled water, W 2 = 1% NaCl + 1% abalone hydrolysate) was detected earlier than the control or the other treatments. In the sensory evaluation, fresh cheese made with the double emulsion showed higher scores for saltiness and overall preference than the control or the other treatments. We concluded that abalone hydrolysate encapsulated in a double emulsion (W 1 is water and W 2 is abalone hydrolysate and NaCl solution) could enhance the saltiness of fresh cheese while maintaining the same salt concentration, without altering its physical properties.

  14. Reactive lysine content in commercially available pet foods

    NARCIS (Netherlands)

    Rooijen, van C.; Bosch, G.; Poel, van der A.F.B.; Wierenga, P.A.; Alexander, L.; Hendriks, W.H.

    2014-01-01

    The Maillard reaction can occur during processing of pet foods. During this reaction, the e-amino group of lysine reacts with reducing sugars to become unavailable for metabolism. The aim of the present study was to determine the reactive lysine (RL; the remaining available lysine) to total lysine

  15. Sperm preparation for ART

    Directory of Open Access Journals (Sweden)

    Schill Wolf-Bernhard

    2003-11-01

    Full Text Available Abstract The onset of clinical assisted reproduction, a quarter of a century ago, required the isolation of motile spermatozoa. As the indication of assisted reproduction shifted from mere gynaecological indications to andrological indications during the years, this urged andrological research to understand the physiology of male germ cell better and develop more sophisticated techniques to separate functional spermatozoa from those that are immotile, have poor morphology or are not capable to fertilize oocytes. Initially, starting from simple washing of spermatozoa, separation techniques, based on different principles like migration, filtration or density gradient centrifugation evolved. The most simple and cheapest is the conventional swim-up procedure. A more sophisticated and most gentle migration method is migration-sedimentation. However, its yield is relatively small and the technique is therefore normally only limited to ejaculates with a high number of motile spermatozoa. Recently, however, the method was also successfully used to isolate spermatozoa for intracytoplasmic sperm injection (ICSI. Sperm separation methods that yield a higher number of motile spermatozoa are glass wool filtration or density gradient centrifugation with different media. Since Percoll® as a density medium was removed from the market in 1996 for clinical use in the human because of its risk of contamination with endotoxins, other media like IxaPrep®, Nycodenz, SilSelect®, PureSperm® or Isolate® were developed in order to replace Percoll®. Today, an array of different methods is available and the selection depends on the quality of the ejaculates, which also includes production of reactive oxygen species (ROS by spermatozoa and leukocytes. Ejaculates with ROS production should not be separated by means of conventional swim-up, as this can severely damage the spermatozoa. In order to protect the male germ cells from the influence of ROS and to stimulate

  16. Genome Sequences of a Novel Picorna-Like Virus from Pacific Abalone (Haliotis discus hannai) in South Korea.

    Science.gov (United States)

    Kim, Jong-Oh; Lee, Jong Kyu; Seo, Yong Bae; Lim, Han-Kyu; Kim, Gun-Do

    2017-09-07

    We report the draft genome sequences of a novel member of the Picornavirales isolated from Pacific abalone ( Haliotis discus hannai ). The full length of the assembled draft genome sequences, obtained by use of a next-generation sequencing technique, were 8,019 nucleotides, including an RNA-dependent RNA polymerase gene (5,088 nucleotides) and a capsid protein gene (2,553 nucleotides). This genome sequence will be useful for understanding viral disease of Pacific abalone. Copyright © 2017 Kim et al.

  17. Relationship between sperm parameters and intracytoplasmic sperm injection outcome

    Directory of Open Access Journals (Sweden)

    Shahla Chaichian

    2015-12-01

    Full Text Available Objectives: With the adventure of intracytoplasmic sperm injection (ICSI technique, great progresses have developed in the treatment of infertility. Concentration on the properties of male’s gamete has been encouraged by the increasing concerns about the causes of ICSI failure. We hence conducted this study to investigate the probable association of sperm parameters with ISCI outcome. Methods: A total of 523 couples referred to Isfahan Fertility and Sterility Center from January 2007 to June 2008 for ICSI. Semen analysis was performed before ICSI procedure according to the WHO criteria. Patients were assigned into successful ICSI (case and failed ICSI (control groups. Sperm parameters were then compared between the 2 groups. Results: One hundred and six patients (20% had successful ICSI results (case group compared with 417 couples (80% with undesirable ICSI outcomes (control group. Among evaluated factors, sperm agglutination (p = 0.007, sperm concentration (p = 0.043, leukocytospermia (p = 0.026 and head abnormality of sperm (p = 0.019 showed statistically significant differences between two groups with differing ICSI results. None of the other semen parameters revealed significant differences between these two groups. Conclusion: Our study showed that some sperm parameters are associated with desirable ICSI outcome. However, it is unclear whether these associations are causal.

  18. Orientation of sea urchin sperms in static magnetic fields: Compared to human sperms

    Science.gov (United States)

    Sakhnini, Lama; Dairi, Maheen

    In this study we report on magnetic orientation of sea urchin and human sperms. The sea urchin and human sperms became oriented parallel to the magnetic field (1 T maximum). The human sperms were totally oriented with magnetic field at about 600 mT. However, the sea urchin sperms show different behavior due to morphological differences between them and the human sperms.

  19. GROW-OUT OF ABALONE Haliotis squamata IN FLOATING CAGES FED DIFFERENT PROPORTIONS OF SEAWEED AND WITH REDUCTION OF STOCKING DENSITY

    Directory of Open Access Journals (Sweden)

    I Nyoman Adiasmara Giri

    2014-06-01

    Full Text Available Abalone is a herbivore marine animal which feeds on seaweed. Abalone culture has a good prospect in terms of price, market share and simple culture technique. Thus, a study was conducted with the aim of finding out an effective and efficient abalone culture technique in terms of feed use and density. In this study, a 42 cm diameter plastic container with a 22 cm height was used. Three vertically arranged containers were used as the experimental group which were put into a net box and hung onto a raft so that the containers were placed in a 4 m depth below the sea surface. The juvenile of abalones being used came from a hatchery production that has been adapted to cages environment with Gracilaria sp. and Ulva sp. feed. The initial density of abalones was 450 for each container, with the initial weight of 2.6-3.2 g and the 2.5-2.7 cm shell lengths. The abalones were fed with Gracilaria sp. and Ulva sp. seaweeds with different Gracilaria sp./Ulva sp. proportions, i.e. 100/0% (A; 80/20% (B; and 60/40% (C as the treatments. Each treatment consisted of two replications. After three months of rearing period, densities of abalones were reduced to be 190 for each experimental unit. Weight and shell length of abalones were measured every month by measuring 25 abalone samples from each experimental unit. The result of the experiment showed that the increase in the Ulva sp. proportion in the feed increased the growth of abalones and decreased the feed conversion. Feeding with Gracilaria sp./Ulva sp. proportion of 60%/40% allowed the best growth of abalones. The decrease of abalone density in the experimental unit after three months of rearing also produced an increase in their growth.

  20. THE EVOLUTION OF SPERM SIZE IN BIRDS.

    Science.gov (United States)

    Briskie, James V; Montgomerie, Robert; Birkhead, Tim R

    1997-06-01

    Sperm size varies enormously among species, but the reasons for this variation remain obscure. Since it has been suggested that swimming velocity increases with sperm length, earlier studies proposed longer (and therefore faster) sperm are advantageous under conditions of intense sperm competition. Nonetheless, previous work has been equivocal, perhaps because the intensity of sperm competition was measured indirectly. DNA profiling now provides a more direct measure of the number of offspring sired by extrapair males, and thus a more direct method of assessing the potential for sperm competition. Using a sample of 21 species of passerine birds for which DNA profiling data were available, we found a positive relation between sperm length and the degree of extrapair paternity. A path analysis, however, revealed that this relationship arises only indirectly through the positive relationship between the rate of extrapair paternity and length of sperm storage tubules (SSTs) in the female. As sperm length is correlated positively with SST length, an increase in the intensity of sperm competition leads to an increase in sperm length only through its effect on SST length. Why females vary SST length with the intensity of sperm competition is not clear, but one possibility is that it increases female control over how sperm are used in fertilization. Males, in turn, may respond on an evolutionary time scale to changes in SST size by increasing sperm length to prevent displacement from rival sperm. Previous theoretical analyses predicting that sperm size should decrease as sperm competition becomes more intense were not supported by our findings. We suggest that future models of sperm-size evolution consider not only the role of sperm competition, but also how female control and manipulation of ejaculates after insemination selects for different sperm morphologies. © 1997 The Society for the Study of Evolution.

  1. Sperm structure and sperm transfer in Pseudopythina subsinuata (Bivalvia, Galeommatoidea)

    DEFF Research Database (Denmark)

    Jespersen, Åse

    2009-01-01

    to the elongate cells. Most females contain one to three "sperm trees", structures consisting of a short stem and numerous branches. They are firmly implanted in the abfrontal part of the gill filament and protrude into the posterior part of the suprabranchial (brooding) chamber. Implantation of the trees causes...... the receptacle and thereupon fuse. A similar process is known in the allied P. tsurumaru, but the resulting structure ("sperm-carrying body") is not attached to the gills....

  2. Myoinositol: does it improve sperm mitochondrial function and sperm motility?

    Science.gov (United States)

    Condorelli, Rosita A; La Vignera, Sandro; Bellanca, Salvatore; Vicari, Enzo; Calogero, Aldo E

    2012-06-01

    To evaluate whether an improvement in mitochondrial membrane potential was associated with sperm motility amelioration and greater sperm recovery after the swim-up procedure. A second purpose was to evaluate the effects of myoinositol (MYO) on sperm apoptosis, quality of chromatin compaction, and DNA integrity. Spermatozoa from 20 normozoospermic men and 20 patients with oligo-astheno-teratozoospermia were incubated in vitro with 2 mg/mL of MYO or phosphate-buffered saline as a control for 2 hours. After this incubation period, sperm motility was evaluated. Flow cytometry was used to analyze the mitochondrial membrane potential, phosphatidylserine externalization, chromatin compactness, and DNA fragmentation. We also evaluated the total number of motile spermatozoa recovered after swim-up after incubation with MYO or phosphate-buffered saline. MYO significantly increased the percentage of spermatozoa with progressive motility in both normozoospermic men and patients with oligo-astheno-teratozoospermia. Motility improvement in the latter group was associated with a significant increase in the percentage of spermatozoa with high mitochondrial membrane potential. MYO had no effects on mitochondrial function in spermatozoa from normozoospermic men. Sperm phosphatidylserine externalization, chromatin compactness, and DNA fragmentation were unaffected by MYO in both groups. After incubation with MYO, the total number of spermatozoa recovered after swim-up had improved significantly in both groups. These data show that MYO increases sperm motility and the number of spermatozoa retrieved after swim-up in both normozoospermic men and patients with abnormal sperm parameters. In patients with oligo-astheno-teratozoospermia, the improvement in these parameters was associated with improved sperm mitochondrial function. These findings support the use of MYO in both in vivo- and in vitro-assisted reproductive techniques. Copyright © 2012 Elsevier Inc. All rights reserved.

  3. Sperm Proteome Maturation in the Mouse Epididymis.

    Science.gov (United States)

    Skerget, Sheri; Rosenow, Matthew A; Petritis, Konstantinos; Karr, Timothy L

    2015-01-01

    In mammals, transit through the epididymis, which involves the acquisition, loss and modification of proteins, is required to confer motility and fertilization competency to sperm. The overall dynamics of maturation is poorly understood, and a systems level understanding of the complex maturation process will provide valuable new information about changes occurring during epididymal transport. We report the proteomes of sperm collected from the caput, corpus and cauda segments of the mouse epididymis, identifying 1536, 1720 and 1234 proteins respectively. This study identified 765 proteins that are present in sperm obtained from all three segments. We identified 1766 proteins that are potentially added (732) or removed (1034) from sperm during epididymal transit. Phenotypic analyses of the caput, corpus and cauda sperm proteomes identified 60 proteins that have known sperm phenotypes when mutated, or absent from sperm. Our analysis indicates that as much as one-third of proteins with known sperm phenotypes are added to sperm during epididymal transit. GO analyses revealed that cauda sperm are enriched for specific functions including sperm-egg recognition and motility, consistent with the observation that sperm acquire motility and fertilization competency during transit through the epididymis. In addition, GO analyses revealed that the immunity protein profile of sperm changes during sperm maturation. Finally, we identified components of the 26S proteasome, the immunoproteasome, and a proteasome activator in mature sperm.

  4. Transport of lysine and hydroxylysine in Streptococcus faecalis.

    Science.gov (United States)

    Friede, J D; Gilboe, D P; Triebwasser, K C; Henderson, L M

    1972-01-01

    Data are presented which support the view that l-lysine is transported by two systems in Streptococcus faecalis. The system with the higher affinity for l-lysine appears to be specific for l-lysine among the common amino acids and to require an energy source. The second system transports both l-lysine and l-arginine and does not appear to require an energy source. Both of these systems will accept hydroxy-l-lysine as a substrate as shown by the energy requirement for hydroxy-l-lysine transport and by the inhibition of uptake by l-arginine as well as by l-lysine. The affinity of both systems appears to be considerably lower for hydroxy-l-lysine than for l-lysine. A mutant of S. faecalis which is resistant to the growth inhibitory action of hydroxy-l-lysine appears to differ from the parent strain by having a defective l-lysine-specific transport system. In this mutant, hydroxy-l-lysine is not readily transported via the l-lysine-specific system because of the mutation or via the second system because of the high concentration of l-arginine present in the growth medium. This overall lack of transport prevents hydroxy-l-lysine from reaching inhibitory levels within the cell.

  5. Amplified fragment length polymorphism analysis to assess crossover interference and homozygosity in gynogenetic diploid Pacific abalone (Haliotis discus hannai).

    Science.gov (United States)

    Nie, H-T; Li, Q; Kong, L-F

    2014-06-01

    Recombination analysis in gynogenetic diploids is a powerful tool for assessing the degree of inbreeding, investigating crossover events and understanding chiasma interference during meiosis. To estimate the marker-centromere recombination rate, the inheritance pattern of 654 amplified fragment length polymorphism (AFLP) markers was examined in the 72-h veliger larvae of two meiogynogenetic diploid families in the Pacific abalone (Haliotis discus hannai). The second-division segregation frequency (y) of the AFLP loci ranged from 0.00 to 0.96, with 23.9% of loci showing y-values higher than 0.67, evidencing the existence of interference. The average recombination frequency across the 654 AFLP loci was 0.45, allowing estimation of the fixation index of 0.55, indicating that meiotic gynogenesis could provide an effective means of rapid inbreeding in the Pacific abalone. The AFLP loci have a small proportion (4.4%) of y-values greater than 0.90, suggesting that a relatively low or intermediate degree of chiasma interference occurred in the abalone chromosomes. The information obtained in this study will enhance our understanding of the abalone genome and will be useful for genetic studies in the species. © 2014 Stichting International Foundation for Animal Genetics.

  6. Effect of drying process assisted by high-pressure impregnation on protein quality and digestibility in red abalone (Haliotis rufescens).

    Science.gov (United States)

    Cepero-Betancourt, Yamira; Oliva-Moresco, Patricio; Pasten-Contreras, Alexis; Tabilo-Munizaga, Gipsy; Pérez-Won, Mario; Moreno-Osorio, Luis; Lemus-Mondaca, Roberto

    2017-10-01

    Abalone (Haliotis spp.) is an exotic seafood product recognized as a protein source of high biological value. Traditional methods used to preserve foods such as drying technology can affect their nutritional quality (protein quality and digestibility). A 28-day rat feeding study was conducted to evaluate the effects of the drying process assisted by high-pressure impregnation (HPI) (350, 450, and 500 MPa × 5 min) on chemical proximate and amino acid compositions and nutritional parameters, such as protein efficiency ratio (PER), true digestibility (TD), net protein ratio, and protein digestibility corrected amino acid score (PDCAAS) of dried abalone. The HPI-assisted drying process ensured excellent protein quality based on PER values, regardless of the pressure level. At 350 and 500 MPa, the HPI-assisted drying process had no negative effect on TD and PDCAAS then, based on nutritional parameters analysed, we recommend HPI-assisted drying process at 350 MPa × 5 min as the best process condition to dry abalone. Variations in nutritional parameters compared to casein protein were observed; nevertheless, the high protein quality and digestibility of HPI-assisted dried abalones were maintained to satisfy the metabolic demands of human beings.

  7. Habitat Preferences of Juvenile Abalone (Haliotis mariae Wood, 1828) Along the Dhofar Coast of Oman and Implications for Conservation

    OpenAIRE

    Schalk Willem Petrus de Waal; Mohammed Balkhair; Ali Al-Mashikhi; Salem Khoom

    2012-01-01

    Surveys were conducted along the eastern Dhofar coast of Oman to investigate densities and habitat preferences of juvenile Haliotis mariae (< 3 cm SL). Average density was 0.62 m-2 (SD 0.56); average urchin density was 3.4 urchins m-2 (SD 3.9). Relationships between juvenile abalone densities and small boulders (

  8. Sperm fractions obtained following density gradient centrifugation in human ejaculates show differences in sperm DNA longevity

    Directory of Open Access Journals (Sweden)

    Jaime Gosálvez

    2014-06-01

    Conclusion: 1 Unnecessary incubation of spermatozoa prior to artificial insemination or in vitro fertilization, should be avoided, since sperm DNA longevity is significantly reduced after ex vivo sperm handling and 2 Although sperm selection by DCG significantly reduces the baseline levels of SDF of sperm in Fraction 3, sperm DNA longevity in this fraction was ultimately lower following 24 h incubation when compared to sperm recovered from non-centrifuged NSS.

  9. Bio-inspired passive actuator simulating an abalone shell mechanism for structural control

    International Nuclear Information System (INIS)

    Yang, Henry T Y; Lin, Chun-Hung; Bridges, Daniel; Randall, Connor J; Hansma, Paul K

    2010-01-01

    An energy dispersion mechanism called 'sacrificial bonds and hidden length', which is found in some biological systems, such as abalone shells and bones, is the inspiration for new strategies for structural control. Sacrificial bonds and hidden length can substantially increase the stiffness and enhance energy dissipation in the constituent molecules of abalone shells and bone. Having been inspired by the usefulness and effectiveness of such a mechanism, which has evolved over millions of years and countless cycles of evolutions, the authors employ the conceptual underpinnings of this mechanism to develop a bio-inspired passive actuator. This paper presents a fundamental method for optimally designing such bio-inspired passive actuators for structural control. To optimize the bio-inspired passive actuator, a simple method utilizing the force–displacement–velocity (FDV) plots based on LQR control is proposed. A linear regression approach is adopted in this research to find the initial values of the desired parameters for the bio-inspired passive actuator. The illustrative examples, conducted by numerical simulation with experimental validation, suggest that the bio-inspired passive actuator based on sacrificial bonds and hidden length may be comparable in performance to state-of-the-art semi-active actuators

  10. Effects of dietary carbohydrates sources on lipids compositions in abalone, Haliotis discus hannai Ino

    Science.gov (United States)

    Wang, Weifang; Mai, Kangsen; Zhang, Wenbing; Xu, Wei; Ai, Qinghui; Yao, Chunfeng; Li, Huitao

    2009-09-01

    A study was conducted to evaluate the effects of dietary carbohydrates on triglyceride, cholesterol and fatty acid concentrations in abalone, Haliotis discus hannai Ino. Six semi-purified diets with different carbohydrates (dextrin, heat-treated wheat starch, wheat starch, corn starch, tapioca starch and potato starch, respectively), all containing a carbohydrate level of 33.5%, were fed to abalone (initial shell length: 29.98 mm ± 0.09 mm; initial weight: 3.42 g ± 0.02 g) for 24 weeks in a recirculation system. The results indicate that serum triglyceride concentrations were significantly ( P starch and wheat starch than those fed with corn starch, and serum cholesterol concentrations were significantly ( P starch than those fed with corn starch. Fatty acid C20:4n-6 in the foot muscles were significantly ( P starch, corn starch, tapioca starch and potato starch. Fatty acid C20:4n-6 in hepatopancreas was significantly ( P starch than those fed with corn starch, tapioca starch and potato starch. Fatty acid C22:6n-3 in the foot muscles were significantly ( P starch than those fed with wheat starch and potato starch.

  11. Nutrient uptake efficiency of Gracilaria chilensis and Ulva lactuca in an IMTA system with the red abalone Haliotis rufescens

    Directory of Open Access Journals (Sweden)

    Juan Macchiavello

    2014-07-01

    Full Text Available The current study examined the nutrient uptake efficiency of Ulva lactuca and Gracilaria chilensis cultivated in tanks associated with the wastewater of a land-based abalone culture. The experiments evaluated different seaweed stocking densities (1200, 1900, 2600, and 3200 g m-2 and water exchange rates (60, 80, 125, and 250 L h-1. The results show that both U. lactuca and G. chilensis were efficient in capturing and removing all of the inorganic nutrients originating from the abalone cultivation for all of the tested conditions. Furthermore, an annual experiment was performed with U. lactuca, cultivated at a stocking density of 1900 g m-2 and at a water exchanged rate of 125 L h-1, in order to evaluate seasonal changes in the nutrient uptake efficiency, productivity, and growth rate associated with the wastewater of a land-based abalone culture. The results confirmed high uptake efficiency during the entire year, equivalent to a 100% removal of the NH4, NO3, and PO4 produced by the land-based abalone culture. The growth rate and productivity of U. lactuca presented a marked seasonality, increasing from fall until summer and varying from 0.5 ± 0.2% to 2.6 ± 0.2% d-1 and 10 ± 6.1% to 73.6 ± 8.4% g m-2 d-1 for sustainable growth rate and productivity, respectively. We conclude that there is sufficient evidence that demonstrates the high possibility of changing the traditional monoculture system of abalone in Chile, to a sustainable integrated multi-trophic aquaculture system, generating positive environmental externalities, including the use of U. lactuca as a biofiltration unit.

  12. Involvement of homocysteine, homocysteine thiolactone, and paraoxonase type 1 (PON-1) in the etiology of defective human sperm function.

    Science.gov (United States)

    Aitken, R J; Flanagan, H M; Connaughton, H; Whiting, S; Hedges, A; Baker, M A

    2016-03-01

    This study reports, for the first time, the significant (p ≤ 0.01) accumulation of homocysteine residues in low density, defective sperm suspensions isolated from patients attending an infertility clinic. This overabundance of homocysteine was not related to a deficiency in folate availability but may have been a reflection of the oxidative stress that characterizes such defective sperm populations. Direct addition of the homocysteine cyclic congener, homocysteine thiolactone, to human spermatozoa resulted in the rapid induction of mitochondrial reactive oxygen species (ROS) generation (p homocysteine molecule was less active and took 24 h to stimulate mitochondrial ROS production possibly because of the need to convert this compound to the corresponding thiolactone before it could exert a measureable biological effect. Thiolactone was also effective in suppressing the carboxymethylation of key proteins in the sperm tail, which are thought to be involved in the regulation of sperm movement. The major enzyme responsible for removing thiolactone from proteins, paraoxonase (PON-1), was shown to be a major target for alkylation by lipid aldehydes, such as 4-hydroxynonenal, generated as a consequence of oxidative stress. Exposure of human spermatozoa to such aldehydes resulted in a dose-dependent accumulation of homocysteine in spermatozoa (p homocysteine thiolactone to interact with the epsilon-amino group of lysine residues on sperm proteins, triggering a raft of significant biological changes in these cells that ultimately compromise sperm function. © 2016 American Society of Andrology and European Academy of Andrology.

  13. Digestible reactive lysine in selected milk-based products.

    Science.gov (United States)

    Rutherfurd, S M; Moughan, P J

    2005-01-01

    Reactive lysine contents, true ileal reactive lysine digestibility, and true ileal digestible reactive lysine contents were determined in a wide range of processed milk products. A previously validated assay based on determining reactive lysine in both food and ileal digesta, after reaction of these materials with O-methylisourea, was applied. Semisynthetic diets containing milk products as the sole sources of protein and including chromic oxide as an indigestible marker were fed to growing rats. Digesta from the terminal ileum were collected posteuthanasia and, with samples of the diets, analyzed for reactive lysine (homoarginine) contents. True reactive lysine digestibility was determined after correcting for endogenous lysine loss at the terminal ileum of rats fed an enzyme hydrolyzed casein-based diet, followed by ultrafiltration (5000 Da) of the digesta. Digestible total lysine (determined using conventional methods) was also determined. The true ileal reactive lysine digestibility was high (>91%) in all the milk products tested, but was highest in the UHT milk (100%) and lowest in the infant formulas (91 to 93%). Total lysine digestibility (conventional measurement) significantly underestimated reactive lysine digestibility for all the products tested. The mean underestimation ranged from 1.3 to 7.1% units. The mean digestible total lysine content was significantly different from the available lysine content for most of the products examined. In some cases this difference was small (milk, whole milk protein, lactose hydrolyzed milk powder, and a sports formula) the difference was greater (6.5 to 14%). This would suggest firstly that total lysine and total lysine digestibility determined using conventional methods were inaccurate when applied to some milk-based foods, and secondly that some of the milk products have undergone lysine modification. In general, milk proteins are a highly digestible source of amino acids and lysine.

  14. Molecular cloning of metal-responsive transcription factor-1 (MTF-1 and transcriptional responses to metal and heat stresses in Pacific abalone, Haliotis discus hannai

    Directory of Open Access Journals (Sweden)

    Sang Yoon Lee

    2017-07-01

    Full Text Available Abstract Background Metal-responsive transcription factor-1 (MTF-1 is a key transcriptional regulator playing crucial roles in metal homeostasis and cellular adaptation to diverse oxidative stresses. In order to understand cellular pathways associated with metal regulation and stress responses in Pacific abalone (Haliotis discus hannai, this study was aimed to isolate the genetic determinant of abalone MTF-1 and to examine its expression characteristics under basal and experimentally stimulated conditions. Results The abalone MTF-1 shared conserved features in zinc-finger DNA binding domain with its orthologs; however, it represented a non-conservative shape in presumed transactivation domain region with the lack of typical motifs for nuclear export signal (NES and Cys-cluster. Abalone MTF-1 promoter exhibited various transcription factor binding motifs that would be potentially related with metal regulation, stress responses, and development. The highest messenger RNA (mRNA expression level of MTF-1 was observed in the testes, and MTF-1 transcripts were detected during the entire period of embryonic and early ontogenic developments. Abalone MTF-1 was found to be Cd inducible and highly modulated by heat shock treatment. Conclusion Abalone MTF-1 possesses a non-consensus structure of activation domains and represents distinct features for its activation mechanism in response to metal overload and heat stress. The activation mechanism of abalone MTF-1 might include both indirect zinc sensing and direct de novo synthesis of transcripts. Taken together, results from this study could be a useful basis for future researches on stress physiology of this abalone species, particularly with regard to heavy metal detoxification and thermal adaptation.

  15. Sperm cleanup and centrifugation processing for cryopreservation.

    Science.gov (United States)

    Sieme, Harald; Oldenhof, Harriëtte

    2015-01-01

    Fertility rates with artificial insemination are highest with good-quality sperm samples. Therefore, nonviable sperm, cellular debris, and seminal plasma are preferably removed from semen samples prior to use or for preservation. Such compounds are sources where reactive oxygen species are generated during storage or upon cryopreservation, impairing sperm function. In this chapter we describe methods to remove seminal plasma and cellular debris from sperm samples, and for selecting morphologically normal motile sperm. The methods that are described here include: ordinary centrifugation, sperm swim-up, glass wool and Sephadex filtration/adherence, and single-layer as well as discontinuous two-layer iodixanol density gradient centrifugation.

  16. Experimental evidence for the effects of polyphenolic compounds from Dictyoneurum californicum Ruprecht (Phaeophyta: Laminariales) on feeding rate and growth in the red abalone Haliotus rufescens Swainson

    Science.gov (United States)

    Winter, Frank C.; Estes, James A.

    1992-01-01

    The effects of polyphenolic compounds from brown algae on grazing and growth rate of the California red abalone Haliotis rufescens Swainson were examined. Abalone consumed three phenolic-poor algal species, Laminaria sinclarii (Harvey) Farlow, Macrocystis pyrifera Agardh, and Nereocystis luetkeana Postels et Ruprecht (mean phenolic content = 0.52% dry mass), at a greater rate than two phenolic-rich species, Dictyoneurum californicum Ruprecht and Cystoseira osmundacea Agardh (mean phenolic content = 4.60% dry mass). This inverse relationship between phenolic content and consumption rate also existed after the algae were macerated and the liquid portion of the blended slurry incorporated in agar discs. However, the correlation between grazing rate and phenolic content imprpve d in this latter experiment, thus suggesting that abalone grazing was deterred significantly by the morphology of L. sinclarii and, to a lesser extent, of M. pyrifera. Polyphenolics extracted from D. californicum reduced abalone grazing rates by 90% when incorporated into agar discs at a concentration of 6 mg·ml−1. Although abalone were unable to maintain body mass when fed ad libitum on macerated M. pyrifera incorporated into agar discs, polyphenolics from D. californicum further inhibited shell growth when added to the discs at 5 mg·ml−1. The abalone ate less of the phenol-containing discs than of the discs lacking phenolics. Our results support findings of several prior studies that polyphenolic compounds from brown algae deter grazing by coastal zone herbivores in the northeast Pacific Ocean.

  17. Effect of dietary lipid on the growth, fatty acid composition and Δ5 Fads expression of abalone ( Haliotis discus hannai Ino) hepatopancreas

    Science.gov (United States)

    Li, Mingzhu; Mai, Kangsen; Ai, Qinghui; He, Gen; Xu, Wei; Zhang, Wenbing; Zhang, Yanjiao; Zhou, Huihui; Liufu, Zhiguo

    2015-04-01

    This study investigated the effect of dietary lipid on the growth, fatty acid composition and Δ5 fatty acyl desaturase genes ( Fads) expression of juvenile abalone ( Haliotis discus hannai Ino) hepatopancreas. Six purified diets were formulated to contain tripalmitin (TP), olive oil (OO, 72.87% 18:1n-9), grape seed oil (GO, 68.67% 18:2n-6), linseed oil (LO, 70.48% 18:3n-3), ARA oil (AO, 41.81% ARA) or EPA oil (EO, 44.09% EPA and 23.67% DAH). No significant difference in survival rate was observed among abalone fed with different diets. Weight gain rate ( WGR) and daily growth rate of shell length ( DGR SL) were significantly increased in abalone fed with diets containing OO, AO and EO, but decreased in abalone fed with LO diet ( P Fads in hepatopancreas of abalone was enhanced by high concentration of 18:3n-3 and suppressed by dietary LC-PUFAs; however it was not affected by dietary high concentration of 18:1n-9 or 18:2n-6. These results provided valuable information for understanding the synthesis of LC-PUFAs and nutritional regulation of Δ5 Fads expression in abalone.

  18. Efficient Production of Enantiopure d-Lysine from l-Lysine by a Two-Enzyme Cascade System

    Directory of Open Access Journals (Sweden)

    Xin Wang

    2016-10-01

    Full Text Available The microbial production of d-lysine has been of great interest as a medicinal raw material. Here, a two-step process for d-lysine production from l-lysine by the successive microbial racemization and asymmetric degradation with lysine racemase and decarboxylase was developed. The whole-cell activities of engineered Escherichia coli expressing racemases from the strains Proteus mirabilis (LYR and Lactobacillus paracasei (AAR were first investigated comparatively. When the strain BL21-LYR with higher racemization activity was employed, l-lysine was rapidly racemized to give dl-lysine, and the d-lysine yield was approximately 48% after 0.5 h. Next, l-lysine was selectively catabolized to generate cadaverine by lysine decarboxylase. The comparative analysis of the decarboxylation activities of resting whole cells, permeabilized cells, and crude enzyme revealed that the crude enzyme was the best biocatalyst for enantiopure d-lysine production. The reaction temperature, pH, metal ion additive, and pyridoxal 5′-phosphate content of this two-step production process were subsequently optimized. Under optimal conditions, 750.7 mmol/L d-lysine was finally obtained from 1710 mmol/L l-lysine after 1 h of racemization reaction and 0.5 h of decarboxylation reaction. d-lysine yield could reach 48.8% with enantiomeric excess (ee ≥ 99%.

  19. Variability in sperm form and function in the context of sperm competition risk in two Tupinambis lizards

    OpenAIRE

    Blengini, Cecilia S; Sergio, Naretto; Gabriela, Cardozo; Giojalas, Laura C; Margarita, Chiaraviglio

    2014-01-01

    In polyandrous species, sperm morphometry and sperm velocity are under strong sexual selection. Although several hypotheses have been proposed to explain the role of sperm competition in sperm trait variation, this aspect is still poorly understood. It has been suggested that an increase in sperm competition pressure could reduce sperm size variation or produce a diversity of sperm to maximize male fertilization success. We aim at elucidating the variability of sperm morphometric traits and v...

  20. Radioactive Lysine in Protein Metabolism Studies

    Science.gov (United States)

    Miller, L. L.; Bale, W. F.; Yuile, C. L.; Masters, R. E.; Tishkoff, G. H.; Whipple,, G. H.

    1950-01-09

    Studies of incorporation of DL-lysine in various body proteins of the dog; the time course of labeled blood proteins; and apparent rate of disappearance of labeled plasma proteins for comparison of behavior of the plasma albumin and globulin fractions; shows more rapid turn over of globulin fraction.

  1. Lysine and arginine requirements of Salminus brasiliensis

    Directory of Open Access Journals (Sweden)

    Jony Koji Dairiki

    2013-08-01

    Full Text Available The objective of this work was to determine the dietary lysine (DL and dietary arginine (DA requirements of dourado (Salminus brasiliensis, through dose-response trials using the amino acid profiles of whole carcasses as a reference. Two experiments were carried out in a completely randomized design (n=4. In the first experiment, groups of 12 feed-conditioned dourado juveniles (11.4±0.2 g were stocked in 60 L cages placed in 300 L plastic indoor tanks in a closed circulation system. Fish were fed for 60 days on diets containing 1.0, 1.5, 2.0, 2.5, 3.0, or 3.5 % dietary lysine. In the second experiment, dourado juveniles (27.0±0.8 g were fed for 60 days on semipurified diets containing arginine at 1.0, 1.5, 2.0, 2.5 or 3.0%, in similar conditions to those of the first experiment. Optimal DL requirements, as determined by broken-line analysis method for final weight, weight gain and specific growth rate, were 2.15% DL or 5% lysine in dietary protein, and 1.48% DA or 3.43% arginine in dietary protein. The best feed conversion ratio is attained with 2.5% DL or 5.8% lysine in dietary protein and 1.4% DA or 3.25% arginine in dietary protein.

  2. Lysine acetylation of major Chlamydia trachomatis antigens

    Directory of Open Access Journals (Sweden)

    Jelena Mihailovic

    2016-03-01

    Our data show that important Ct antigens could be post-translationally modified by acetylation of lysine residues at multiple sites. Further studies are needed to investigate total acetylome of Ct and the impact PTMs might have on Ct biology and pathogenicity.

  3. Immunomodulatory activity of chicken NK-lysin peptides

    Science.gov (United States)

    Chicken NK-lysin (cNK-lysin), the chicken homologue of human granulysin, is a cationic amphiphilic antimicrobial peptide (AMP) produced by cytotoxic T cells and natural killer cells. We have previously demonstrated that cNK-lysin and cNK-2, which is a synthetic peptide incorporating core alpha-helic...

  4. Female sperm storage in reptiles.

    Science.gov (United States)

    Sever, David M; Hamlett, William C

    2002-02-01

    Internal fertilization and oviparity most likely are symplesiomorphies for modern reptiles, and viviparity has evolved independently numerous times in Sauria and Serpentes. Oviducal sperm storage is known in females of all taxa except Amphisbaenia. However, in Rhynchocephalia and Crocodilia, sperm storage is poorly studied, and specialized sperm storage tubules (Ssts) are unknown. We use the molecular phylogenetic hypothesis [(Chelonia+Archosauria) (Squamata)] to trace evolution of sperm storage characters. Ssts arose independently in Chelonia and Squamata. Turtles possess albumen-secreting glands in the anterior half of the oviduct (the tuba or isthmus), and the most distal of these glands also serve as Ssts; in addition, some turtles possess Ssts in the adjacent segment of the oviduct, the uterus. Squamates lack albumen-secreting glands, and the ancestral state is possession of Ssts in the posterior infundibulum (uterine tube). Secondarily, iguanids have evolved vaginal Ssts. In this paper, we present the first ultrastructural observations on vaginal Ssts in lizards, using Anolis sagrei (Polychrotidae). Proximally, the neck of these simple tubular glands continues the alternation of ciliated and secretory cells lining the lumen of the vagina. However, the epithelial cells of the distal sperm storage area are neither secretory nor ciliated. The Ssts of Anolis are more similar to those of birds more than to infundibular receptacles in snakes and lizards. Copyright 2002 Wiley-Liss, Inc.

  5. Psychosocial counselling of identifiable sperm donors

    NARCIS (Netherlands)

    Visser, M.; Mochtar, M.H.; de Melker, A.A.; van der Veen, F.; Repping, S.; Gerrits, T.

    2016-01-01

    STUDY QUESTION: What do identifiable sperm donors feel about psychosocial counselling? SUMMARY ANSWER: Identifiable sperm donors found it important that psychosocial counselling focused on emotional consequences and on rules and regulations and they expected to have access to psychosocial

  6. Psychosocial counselling of identifiable sperm donors

    NARCIS (Netherlands)

    Visser, M. [=Marja; Mochtar, M. H.; de Melker, A. A.; van der Veen, F.; Repping, S.; Gerrits, T.

    2016-01-01

    What do identifiable sperm donors feel about psychosocial counselling? Identifiable sperm donors found it important that psychosocial counselling focused on emotional consequences and on rules and regulations and they expected to have access to psychosocial counselling at the time that

  7. Variation in sperm morphology among Afrotropical sunbirds

    Czech Academy of Sciences Publication Activity Database

    Omotoriogun, T. C.; Laskemoen, T.; Rowe, M.; Albrecht, Tomáš; Bowie, R. C. K.; Sedláček, O.; Hořák, D.; Ottosson, U.; Lifjeld, J. T.

    2016-01-01

    Roč. 158, č. 1 (2016), s. 155-166 ISSN 0019-1019 Institutional support: RVO:68081766 Keywords : comparative analysis * Nectariniidae * phylogenetic signal * sperm competition * sperm size Subject RIV: EG - Zoology Impact factor: 2.279, year: 2016

  8. Antibacterial activity and immune responses of a molluscan macrophage expressed gene-1 from disk abalone, Haliotis discus discus.

    Science.gov (United States)

    Bathige, S D N K; Umasuthan, Navaneethaiyer; Whang, Ilson; Lim, Bong-Soo; Won, Seung Hwan; Lee, Jehee

    2014-08-01

    The membrane-attack complex/perforin (MACPF) domain-containing proteins play an important role in the innate immune response against invading microbial pathogens. In the current study, a member of the MACPF domain-containing proteins, macrophage expressed gene-1 (MPEG1) encoding 730 amino acids with the theoretical molecular mass of 79.6 kDa and an isoelectric point (pI) of 6.49 was characterized from disk abalone Haliotis discus discus (AbMPEG1). We found that the characteristic MACPF domain (Val(131)-Tyr(348)) and transmembrane segment (Ala(669)-Ile(691)) of AbMPEG1 are located in the N- and C-terminal ends of the protein, respectively. Ortholog comparison revealed that AbMPEG1 has the highest sequence identity with its pink abalone counterpart, while sequences identities of greater than 90% were observed with MPEG1 members from other abalone species. Likewise, the furin cleavage site KRRRK was highly conserved in all abalone species, but not in other species investigated. We identified an intron-less genomic sequence within disk abalone AbMPEG1, which was similar to other mammalian, avian, and reptilian counterparts. Transcription factor binding sites, which are important for immune responses, were identified in the 5'-flanking region of AbMPEG1. qPCR revealed AbMPEG1 transcripts are present in every tissues examined, with the highest expression level occurring in mantle tissue. Significant up-regulation of AbMPEG1 transcript levels was observed in hemocytes and gill tissues following challenges with pathogens (Vibrio parahemolyticus, Listeria monocytogenes and viral hemorrhagic septicemia virus) as well as pathogen-associated molecular patterns (PAMPs: lipopolysaccharides and poly I:C immunostimulant). Finally, the antibacterial activity of the MACPF domain was characterized against Gram-negative and -positive bacteria using a recombinant peptide. Taken together, these results indicate that the biological significance of the AbMPEG1 gene includes a role in

  9. Boundaries of Ethics, Sperm on the Border –The Globalization of Danish Sperm

    DEFF Research Database (Denmark)

    Willum Adrian, Stine

    During the past two decades, Denmark has developed into an important destination for fertility travelers in need of donor sperm. A variety of different clinics providing sperm with different modes of clinical care have developed, and an increasing number of their customers come from abroad. One...... of the reasons why Danish fertility clinics have enabled women from Denmark and abroad to turn to sperm donation is, that two of the largest sperm banks in Europe are Danish. In contrast to many other countries where sperm is in demand, the Danish sperm banks are able to provide enough sperm for the national...... market as well as clinics worldwide. Furthermore, one of the sperm banks market sperm for self-insemination and deliver it to private customers by UPS. Although sperm donation is an old and low technology compared to other reproductive technologies, this development has taken place at the same time...

  10. Bioenergetics of mammalian sperm capacitation.

    Science.gov (United States)

    Ferramosca, Alessandra; Zara, Vincenzo

    2014-01-01

    After ejaculation, the mammalian male gamete must undergo the capacitation process, which is a prerequisite for egg fertilization. The bioenergetics of sperm capacitation is poorly understood despite its fundamental role in sustaining the biochemical and molecular events occurring during gamete activation. Glycolysis and mitochondrial oxidative phosphorylation (OXPHOS) are the two major metabolic pathways producing ATP which is the primary source of energy for spermatozoa. Since recent data suggest that spermatozoa have the ability to use different metabolic substrates, the main aim of this work is to present a broad overview of the current knowledge on the energy-producing metabolic pathways operating inside sperm mitochondria during capacitation in different mammalian species. Metabolism of glucose and of other energetic substrates, such as pyruvate, lactate, and citrate, is critically analyzed. Such knowledge, besides its obvious importance for basic science, could eventually translate into the development of novel strategies for treatment of male infertility, artificial reproduction, and sperm selection methods.

  11. Microfluidic single sperm entrapment and analysis

    NARCIS (Netherlands)

    de Wagenaar, B.; Berendsen, Johanna Theodora Wilhelmina; Berendsen, J.T.W.; Bomer, Johan G.; Olthuis, Wouter; van den Berg, Albert; Segerink, Loes Irene

    2015-01-01

    Selection of healthy spermatozoa is of crucial importance for the success rates of assisted reproduction technologies (ART) such as in vitro fertilization and intra-cytoplasmic sperm injection. Although sperm selection for ART procedures is predominantly based on sperm motility, successful

  12. Genetic characterization of five hatchery populations of the Pacific Abalone (Haliotis discus hannai) using microsatellite markers.

    Science.gov (United States)

    An, Hye Suck; Lee, Jang Wook; Kim, Hyun Chul; Myeong, Jeong-In

    2011-01-01

    The Pacific abalone, Haliotis discus hannai, is a popular food in Eastern Asia. Aquacultural production of this species has increased because of recent resource declines, the growing consumption, and ongoing government-operated stock release programs. Therefore, the genetic characterization of hatchery populations is necessary to maintain the genetic diversity of this species and to develop more effective aquaculture practices. We analyzed the genetic structures of five cultured populations in Korea using six microsatellite markers. The number of alleles per locus ranged from 15 to 64, with an average of 23.5. The mean observed and expected heterozygosities were 0.797 and 0.904, respectively. The inbreeding coefficient F(IS) ranged from 0.054 to 0.184 (mean F(IS) = 0.121 ± 0.056). The genetic differentiation across all populations was low but significant (overall F(ST) = 0.009, P discus hannai fisheries and the aquaculture industry.

  13. The combination between cations and sulfated polysaccharide from abalone gonad (Haliotis discus hannai Ino).

    Science.gov (United States)

    Guo, Yiming; Cong, Shuang; Zhao, Jun; Dong, Yingying; Li, Tingting; Zhu, Beiwei; Song, Shuang; Wen, Chengrong

    2018-05-15

    Effects of cations (Na + , K + , Ca 2+ , Mg 2+ ) on the structure of abalone gonad sulfated polysaccharide (AGSP) were studied by means of Congo red test, X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR) and scanning electron microscope (SEM). The results showed that the local ordered helical conformation was observed in AGSP. The cations could combine with anionic groups in AGSP by ionic bonds, which caused the weakening of electrostatic repulsion and crosslinking of molecular chains. Furthermore, the effects of divalent cations on the conformation were more obvious than that of monovalent cations, and divalent cations led to the aggregation of AGSP due to the more interaction sites. In short, AGSP molecular chains were crosslinked through ionic bonds after adding cations, and, divalent cations could induce the aggregation of AGSP by electrostatic interactions. This study will provide valuable insights for the further research on AGSP conformation. Copyright © 2018 Elsevier Ltd. All rights reserved.

  14. Antimicrobial peptide, hdMolluscidin, purified from the gill of the abalone, Haliotis discus.

    Science.gov (United States)

    Seo, Jung-Kil; Go, Hye-Jin; Kim, Chan-Hee; Nam, Bo-Hye; Park, Nam Gyu

    2016-05-01

    A 4.7 kDa antimicrobial peptide was purified from the acidified gill extract of the Abalone, Haliotis discus, by cation-exchange and C18 reversed-phase high performance liquid chromatography (HPLC). Comparison of the amino acid sequences and molecular weight of this peptide with those of other known antimicrobial peptides revealed that this antimicrobial peptide have high sequence homology with that of cgMolluscidin and was designated hdMolluscidin. hdMolluscidin is composed of 46 amino acid residues containing several dibasic residue repeats like KK or K-R. hdMolluscidin showed potent antimicrobial activity against both Gram-positive bacteria including Bacillus subtilis and Staphylococcus aureus (minimal effective concentrations [MECs]; 0.8-19.0 μg/mL) and Gram-negative bacteria including Aeromonas hydrophila, Escherichia coli, Pseudomonas aeruginosa, Salmonella enterica, Shigella flexneri, and Vibrio parahemolyticus ([MECs]; 1.0-4.0 μg/mL) without hemolytic activity. However, hdMolluscidin did not show any significant activity against Candida albicans. The secondary structural prediction suggested that hdMolluscidin might not form an ordered or an amphipathic structure. hdMolluscidin did not show membrane permeabilization or leakage ability. The full-length hdMolluscidin cDNA contained 566-bp, including a 5'-untranslated region (UTR) of 63-bp, a 3'-UTR of 359-bp, and an open reading frame of 144-bp encoding 47 amino acids (containing Met). cDNA study of hdMolluscidin suggests that it is expressed as a mature peptide. Our results indicate that hdMolluscidin could relate to the innate immune defenses in abalone and it may not act directly on bacterial membrane. Copyright © 2016 Elsevier Ltd. All rights reserved.

  15. Characterization and spatiotemporal expression of gonadotropin-releasing hormone in the Pacific abalone, Haliotis discus hannai.

    Science.gov (United States)

    Kim, Tae Ha; Kim, Mi Ae; Kim, Kyeong Seop; Kim, Jae Won; Lim, Han Kyu; Lee, Jung Sick; Sohn, Young Chang

    2017-07-01

    Gonadotropin-releasing hormone (GnRH) is a key neuropeptide regulating reproduction in humans and other vertebrates. Recently, GnRH-like cDNAs and peptides were reported in marine mollusks, implying that GnRH-mediated reproduction is an ancient neuroendocrine system that arose prior to the divergence of protostomes and deuterostomes. Here, we evaluated the reproductive control system mediated by GnRH in the Pacific abalone Haliotis discus hannai. We cloned a prepro-GnRH cDNA (Hdh-GnRH) from the pleural-pedal ganglion (PPG) in H. discus hannai, and analyzed its spatiotemporal gene expression pattern. The open reading frame of Hdh-GnRH encodes a protein of 101 amino acids, consisting of a signal peptide, a GnRH dodecapeptide, a cleavage site, and a GnRH-associated peptide. This structure and sequence are highly similar to GnRH-like peptides reported for mollusks and other invertebrates. Quantitative polymerase chain reaction demonstrated that Hdh-GnRH mRNA was more strongly expressed in the ganglions (PPG and cerebral ganglion [CG]) than in other tissues (gonads, gills, intestine, hemocytes, muscle, and mantle) in both sexes. In females, the expression levels of Hdh-GnRH mRNA in the PPG and branchial ganglion (BG) were significantly higher at the ripe and partial spent stages than at the early and late active stages. In males, Hdh-GnRH mRNA levels in the BG showed a significant increase in the partial spent stage. Unexpectedly, Hdh-GnRH levels in the CG were not significantly different among the examined stages in both sexes. These results suggest that Hdh-GnRH mRNA expression profiles in the BG and possibly the PPG are tightly correlated with abalone reproductive activities. Copyright © 2017 Elsevier Inc. All rights reserved.

  16. Solea senegalensis sperm cryopreservation: New insights on sperm quality.

    Directory of Open Access Journals (Sweden)

    Marta F Riesco

    Full Text Available Cryopreservation of Senegalese sole sperm can represent an alternative to overcome some reproductive problems of this species. However, it is important to guarantee the safe use of cryopreserved sperm by selecting an appropriate protocol according to a high demand quality need to be ensured. It has been demonstrated that traditional assays such as motility and viability do not provide enough information to identify specific damage caused by cryopreservation process (freezing and thawing. Specific tests, including lipid peroxidation and DNA damage, should be performed. In the present study, motility and lipid peroxidation were performed as specific tests allowing us to discard cryopreservation conditions such as methanol as internal cryoprotectant and bovine serum albumin as external cryoprotectant. In addition, a caspase 3/7 detection by flow cytometry was performed to analyze apoptosis activity in the best selected conditions. Moreover, new highly sensitive tests based on transcript number detection have recently been described in fish sperm cryopreservation. For this reason, a transcript level detection assay was performed on certain oxidative and chaperone genes related to fertilization ability and embryo development (hsp70, hsp90BB, hsp90AA, gpx to select the best cryopreservation conditions. DMSO+ egg yolk proved to be the best cryoprotectant combination in terms of transcript level. This study describes an optimized cryopreservation protocol for Solea senegalensis sperm demonstrating for the first time that transcript degradation is the most sensitive predictor of cell status in this species after cryopreservation.

  17. Global analysis of protein lysine succinylation profiles in common wheat.

    Science.gov (United States)

    Zhang, Yumei; Wang, Guangyuan; Song, Limin; Mu, Ping; Wang, Shu; Liang, Wenxing; Lin, Qi

    2017-04-20

    Protein lysine succinylation is an important post-translational modification and plays a critical regulatory role in almost every aspects of cell metabolism in both eukaryotes and prokaryotes. Common wheat is one of the major global cereal crops. However, to date, little is known about the functions of lysine succinylation in this plant. Here, we performed a global analysis of lysine succinylation in wheat and examined its overlap with lysine acetylation. In total, 330 lysine succinylated modification sites were identified in 173 proteins. Bioinformatics analysis showed that the modified proteins are distributed in multiple subcellular compartments and are involved in a wide variety of biological processes such as photosynthesis and the Calvin-Benson cycle, suggesting an important role for lysine succinylation in these processes. Five putative succinylation motifs were identified. A protein interaction network analysis revealed that diverse interactions are modulated by protein succinylation. Moreover, 21 succinyl-lysine sites were found to be acetylated at the same position, and 33 proteins were modified by both acetylation and succinylation, suggesting an extensive overlap between succinylation and acetylation in common wheat. Comparative analysis indicated that lysine succinylation is conserved between common wheat and Brachypodium distachyon. These results suggest that lysine succinylation is involved in diverse biological processes, especially in photosynthesis and carbon fixation. This systematic analysis represents the first global analysis of lysine succinylation in common wheat and provides an important resource for exploring the physiological role of lysine succinylation in this cereal crop and likely in all plants.

  18. Bacterial Lysine Decarboxylase Influences Human Dental Biofilm Lysine Content, Biofilm Accumulation and Sub-Clinical Gingival Inflammation

    Science.gov (United States)

    Lohinai, Z.; Keremi, B.; Szoko, E.; Tabi, T.; Szabo, C.; Tulassay, Z.; Levine, M.

    2012-01-01

    Background Dental biofilms contain a protein that inhibits mammalian cell growth, possibly lysine decarboxylase from Eikenella corrodens. This enzyme decarboxylates lysine, an essential amino acid for dentally attached cell turnover in gingival sulci. Lysine depletion may stop this turnover, impairing the barrier to bacterial compounds. The aims of this study were to determine biofilm lysine and cadaverine contents before oral hygiene restriction (OHR), and their association with plaque index (PI) and gingival crevicular fluid (GCF) after OHR for a week. Methods Laser-induced fluorescence after capillary electrophoresis was used to determine lysine and cadaverine contents in dental biofilm, tongue biofilm and saliva before OHR and in dental biofilm after OHR. Results Before OHR, lysine and cadaverine contents of dental biofilm were similar and 10-fold greater than in saliva or tongue biofilm. After a week of OHR, the biofilm content of cadaverine increased and that of lysine decreased, consistent with greater biofilm lysine decarboxylase activity. Regression indicated that PI and GCF exudation were positively related to biofilm lysine post-OHR, unless biofilm lysine exceeded the minimal blood plasma content in which case PI was further increased but GCF exudation was reduced. Conclusions After OHR, lysine decarboxylase activity seems to determine biofilm lysine content and biofilm accumulation. When biofilm lysine exceeds minimal blood plasma content after OHR, less GCF appeared despite more biofilm. Lysine appears important for biofilm accumulation and the epithelial barrier to bacterial proinflammatory agents. Clinical Relevance Inhibiting lysine decarboxylase may retard the increased GCF exudation required for microbial development and gingivitis. PMID:22141361

  19. No evidence for sperm priming responses under varying sperm competition risk or intensity in guppies

    Science.gov (United States)

    Evans, Jonathan P.

    2009-07-01

    Sperm competition theory predicts that males should tailor their investment in ejaculates according to the number of rival males competing to fertilize a female’s eggs. Research spanning several taxa supports this prediction by showing that males are often sensitive to the level of sperm competition and adjust their investment in sperm numbers accordingly. More recent work has revealed that males may also tailor the quality of sperm according to the number of males competing for fertilization. Here I test for both effects in guppies ( Poecilia reticulata) in an experiment that simultaneously evaluates the risk and intensity models of sperm competition. The experiment determined whether male guppies adjust the number (stripped ejaculate size) and quality (sperm velocity and viability) of sperm that are primed over a 3-day period according to experimental changes in the perceived level of sperm competition. A total of 136 focal males were initially stripped of all retrievable sperm and assayed for these sperm traits before being allocated at random to one of four treatments simulating different levels of sperm competition risk and intensity. During the 3-day treatment phase, focal males had visual and olfactory access to a sexually receptive (initially virgin) female maintained with different numbers of stimulus males to simulate variation in the risk and intensity of sperm competition. Following this, males were assayed again for the sperm traits. Contrary to predictions, there was no significant change in any of the measured variables among treatments, although qualitatively the patterns for sperm velocity and viability did conform to expectation. The lack of any trend for the number of sperm primed was unequivocal and future work examining the effects of sperm competition on sperm production should focus on whether males differentially allocate sperm numbers among matings that differ in the level of sperm competition.

  20. Allograft inflammatory factor-1 in disk abalone (Haliotis discus discus): molecular cloning, transcriptional regulation against immune challenge and tissue injury.

    Science.gov (United States)

    De Zoysa, Mahanama; Nikapitiya, Chamilani; Kim, Yucheol; Oh, Chulhong; Kang, Do-Hyung; Whang, Ilson; Kim, Se-Jae; Lee, Jae-Seong; Choi, Cheol Young; Lee, Jehee

    2010-08-01

    Here, we report the identification and characterization of allograft inflammatory factor-1 (AIF-1) from disk abalone Haliotis discus discus that was denoted as AbAIF-1. The full-length cDNA of AbAIF-1 consists of a coding region (453 bp) for 151 amino acids with a 17 kDa molecular mass. Analysis of AbAIF-1 sequence showed that it shares characteristic two EF hand Ca(+2)-binding motifs. Results from phylogenetic analysis further confirm that AbAIF-1 is a member of the AIF-1 family similar to invertebrate and vertebrate counterparts suggesting it has high evolutional conservation. Tissue-specific expression and transcriptional regulation of AbAIF-1 were analyzed after bacteria (Vibrio alginolyticus, Vibrio parahemolyticus and Lysteria monocytogenes), viral hemorrhagic septicemia virus (VHSV) immune challenge and during tissue injury by quantitative real-time PCR. It is shown that the expression of AbAIF-1 mRNA was expressed ubiquitously in all selected tissues in constitutive manner showing the highest level in hemocytes. Upon bacteria and VHSV challenge, AbAIF-1 showed the significant up-regulation in hemocytes than gills. After the tissue injury in shell and mantle, AbAIF-1 and antioxidant selenium-dependant glutathione peroxidase (SeGPx) transcripts were significantly upregulated in abalone hemocytes. Taken together, these findings suggest that AIF-1 could response against the pathogenic challenge or tissue injury in abalone like mollusks. Also, AbAIF-1 may involve in wound healing and shell repair after the tissue injury of abalone. Copyright 2010 Elsevier Ltd. All rights reserved.

  1. Transcriptional up-regulation of a novel ferritin homolog in abalone Haliotis discus hannai Ino by dietary iron.

    Science.gov (United States)

    Wu, Chenglong; Zhang, Wenbing; Mai, Kangsen; Xu, Wei; Wang, Xiaojie; Ma, Hongming; Liufu, Zhiguo

    2010-11-01

    A novel cDNA encoding ferritin (HdhNFT) was cloned from the hepatopancreas of abalone, Haliotis discus hannai Ino. The deduced protein contains 171 amino acid residues with a predicted molecular mass (MW) about 19.8 kDa and theoretical isoelectric point (pI) of 4.792. Amino acid alignment revealed that HdhNFT shared high similarity with other known ferritins. The HdhNFT contained a highly conserved motif for the ferroxidase center, which consists of seven residues of a typical vertebrate heavy-chain ferritin with a typical stem-loop structure. HdhNFT mRNA contains a 27 bp iron-responsive element (IRE) in the 5'-untranslated region. This IRE exhibited 82.14% similarity with abalone H. discus discus and 78.57% similarity with Pacific oyster Crassostrea gigas IREs. By real-time PCR assays, the mRNA transcripts of HdhNFT were found to be higher expressed in kidney, hepatopancreas, gill, mantle and muscle than in haemocytes and gonad. Moreover, mRNA expression levels of HdhNFT in the hepatopancreas and haemocytes were measured by real-time PCR in abalone fed with graded levels of dietary iron (29.2, 65.7, 1267.2 and 6264.7 mg/kg). Results showed that the expression of the HdhNFT mRNA increased with dietary iron contents. Furthermore, the maximum value of the HdhNFT mRNA was found in the treatment with 6264.7 mg/kg of dietary iron. These data indicated that dietary iron can up-regulate HdhNFT at transcriptional level in abalone. Copyright © 2010 Elsevier Inc. All rights reserved.

  2. Effects of temperature and salinity on survival, growth and DNA methylation of juvenile Pacific abalone, Haliotis discus hannai Ino

    Science.gov (United States)

    Kong, Ning; Liu, Xiao; Li, Junyuan; Mu, Wendan; Lian, Jianwu; Xue, Yanjie; Li, Qi

    2017-09-01

    Temperature and salinity are two of the most potent abiotic factors influencing marine mollusks. In this study, we investigated the individual and combined effects of temperature and salinity on the survival and growth of juvenile Pacific abalone, Haliotis discus hannai Ino, and also examined the DNA methylation alteration that may underpin the phenotypic variation of abalone exposed to different rearing conditions. The single-factor data showed that the suitable ranges of temperature and salinity were 16-28°C at a constant salinity of 32, and 24-40 at a constant temperature of 20°C, respectively. The two-factor data indicated that both survival and growth were significantly affected by temperature, salinity and their interaction. The optimal temperature-salinity combination for juveniles was 23-25°C and 30-36. To explore environment-induced DNA methylation alteration, the methylation-sensitive amplified polymorphism (MSAP) technique was used to analyze the genomic methylation profiles of abalone reared in optimal and adverse conditions. Neither temperature nor salinity induced evident changes in the global methylation level, but 67 and 63 differentially methylated loci were identified in temperature and salinity treatments, respectively. The between-group eigen analysis also showed that both temperature and salinity could induce epigenetic differentiation in H. discus hannai Ino. The results of our study provide optimal rearing conditions for juvenile H. discus hannai Ino, and represent the first step toward revealing the epigenetic regulatory mechanism of abalone in response to thermal and salt stresses.

  3. Histone Lysine Methylation and Neurodevelopmental Disorders

    Directory of Open Access Journals (Sweden)

    Jeong-Hoon Kim

    2017-06-01

    Full Text Available Methylation of several lysine residues of histones is a crucial mechanism for relatively long-term regulation of genomic activity. Recent molecular biological studies have demonstrated that the function of histone methylation is more diverse and complex than previously thought. Moreover, studies using newly available genomics techniques, such as exome sequencing, have identified an increasing number of histone lysine methylation-related genes as intellectual disability-associated genes, which highlights the importance of accurate control of histone methylation during neurogenesis. However, given the functional diversity and complexity of histone methylation within the cell, the study of the molecular basis of histone methylation-related neurodevelopmental disorders is currently still in its infancy. Here, we review the latest studies that revealed the pathological implications of alterations in histone methylation status in the context of various neurodevelopmental disorders and propose possible therapeutic application of epigenetic compounds regulating histone methylation status for the treatment of these diseases.

  4. Optimization of Direct Lysine Decarboxylase Biotransformation for Cadaverine Production with Whole-Cell Biocatalysts at High Lysine Concentration.

    Science.gov (United States)

    Kim, Hyun Joong; Kim, Yong Hyun; Shin, Ji-Hyun; Bhatia, Shashi Kant; Sathiyanarayanan, Ganesan; Seo, Hyung-Min; Choi, Kwon Young; Yang, Yung-Hun; Park, Kyungmoon

    2015-07-01

    Cadaverine (1,5-diaminopentane) is an important industrial chemical with a wide range of applications. Although there have been many efforts to produce cadaverine through fermentation, there are not many reports of the direct cadaverine production from lysine using biotransformation. Whole-cell reactions were examined using a recombinant Escherichia coli strain overexpressing the E. coli MG1655 cadA gene, and various parameters were investigated for the whole-cell bioconversion of lysine to cadaverine. A high concentration of lysine resulted in the synthesis of pyridoxal-5'-phosphate (PLP) and it was found to be a critical control factor for the biotransformation of lysine to cadaverine. When 0.025 mM PLP and 1.75 M lysine in 500 mM sodium acetate buffer (pH6) were used, consumption of 91% lysine and conversion of about 80% lysine to cadaverine were successfully achieved.

  5. Cat fertilization by mouse sperm injection.

    Science.gov (United States)

    Jin, Yong-Xun; Cui, Xiang-Shun; Yu, Xian-Feng; Lee, Sung-Hyun; Wang, Qing-Ling; Gao, Wei-Wei; Xu, Yong-Nan; Sun, Shao-Chen; Kong, Il-Keun; Kim, Nam-Hyung

    2012-11-01

    Interspecies intracytoplasmic sperm injection has been carried out to understand species-specific differences in oocyte environments and sperm components during fertilization. While sperm aster organization during cat fertilization requires a paternally derived centriole, mouse and hamster fertilization occur within the maternal centrosomal components. To address the questions of where sperm aster assembly occurs and whether complete fertilization is achieved in cat oocytes by interspecies sperm, we studied the fertilization processes of cat oocytes following the injection of cat, mouse, or hamster sperm. Male and female pronuclear formations were not different in the cat oocytes at 6 h following cat, mouse or hamster sperm injection. Microtubule asters were seen in all oocytes following intracytoplasmic injection of cat, mouse or hamster sperm. Immunocytochemical staining with a histone H3-m2K9 antibody revealed that mouse sperm chromatin is incorporated normally with cat egg chromatin, and that the cat eggs fertilized with mouse sperm enter metaphase and become normal 2-cell stage embryos. These results suggest that sperm aster formation is maternally dependent, and that fertilization processes and cleavage occur in a non-species specific manner in cat oocytes.

  6. Etiologies of sperm oxidative stress

    Directory of Open Access Journals (Sweden)

    Parvin Sabeti

    2016-04-01

    Full Text Available Sperm is particularly susceptible to reactive oxygen species (ROS during critical phases of spermiogenesis. However, the level of seminal ROS is restricted by seminal antioxidants which have beneficial effects on sperm parameters and developmental potentials. Mitochondria and sperm plasma membrane are two major sites of ROS generation in sperm cells. Besides, leukocytes including polymer phonuclear (PMN leukocytes and macrophages produce broad category of molecules including oxygen free radicals, non-radical species and reactive nitrogen species. Physiological role of ROS increase the intracellular cAMP which then activate protein kinase in male reproductive system. This indicates that spermatozoa need small amounts of ROS to acquire the ability of nuclear maturation regulation and condensation to fertilize the oocyte. There is a long list of intrinsic and extrinsic factors which can induce oxidative stress to interact with lipids, proteins and DNA molecules. As a result, we have lipid peroxidation, DNA fragmentation, axonemal damage, denaturation of the enzymes, over generation of superoxide in the mitochondria, lower antioxidant activity and finally abnormal spermatogenesis. If oxidative stress is considered as one of the main cause of DNA damage in the germ cells, then there should be good reason for antioxidant therapy in these conditions

  7. Proteomic profiling of eggs from a hybrid abalone and its parental lines: Haliotis discus hannai Ino and Haliotis gigantea.

    Science.gov (United States)

    Di, Guilan; Luo, Xuan; Huang, Miaoqin; Chen, Jun; Kong, Xianghui; Miao, Xiulian; Ke, Caihuan

    2015-12-01

    Proteomic analysis was performed on the eggs of hybrid abalone and their corresponding parental lines. A total of 915 ± 19 stained protein spots were detected from Haliotis discus hannai♀ × H. discus hannai♂ (DD), 935 ± 16 from H. gigantea♀ × H. gigantea♂ (GG) and 923 ± 13 from H. gigantea♀ × H. discus hannai♂ (GD). The spots from DD and GD were clustered together. The distance between DD and GG was maximal by hierarchical cluster analysis. A total of 112 protein gel spots were identified; of these, 59 were abalone proteins. The proteins were involved in major biological processes including energy metabolism, proliferation, apoptosis, signal transduction, immunity, lipid metabolism, electron carrier proteins, protein biosynthesis and decomposition, and cytoskeletal structure. Three of 20 differential expression protein spots involved in energy metabolism exhibited as upregulated in GD, 13 spots exhibited additivity, and four spots exhibited as downregulated in the offspring. Eleven protein spots were expressed at the highest level in DD. The proteins involved in stress responses included superoxide dismutase, peroxiredoxin 6, thioredoxin peroxidase and glutathione-S-transferase. Two of seven differential expression protein spots involved in response to stress exhibited as upregulated in GD, three exhibited additivity, and two exhibited as downregulated. These results might suggest that proteomic approaches are suitable for the analysis of hybrids and the functional prediction of abalone hybridization. © 2015 Stichting International Foundation for Animal Genetics.

  8. BRICHOS domain-containing leukocyte cell-derived chemotaxin 1-like cDNA from disk abalone Haliotis discus discus.

    Science.gov (United States)

    Kim, Yucheol; De Zoysa, Mahanama; Lee, Youngdeuk; Whang, Ilson; Lee, Jehee

    2010-11-01

    A BRICHOS domain-containing leukocyte cell-derived chemotaxin 1-like cDNA was cloned from the disk abalone (Haliotis discus discus) and designated as AbLECT-1. A full-length (705 bp) of AbLECT-1 cDNA was composed of a 576 bp open reading frame that translates into a putative peptide of 192 amino acids. Deduced amino acid sequence of AbLECT-1 had 15.5- and 27.8% identity and similarity to human LECT-1, respectively. Quantitative real-time PCR analysis results showed that the mRNA of AbLECT-1 was constitutively expressed in abalone hemocytes, gills, mantle, muscle, digestive tract and hepatopancreas in a tissue-specific manner. Moreover, the AbLECT-1 transcription level was induced in hemocytes after challenge with Vibrio alginolyticus, Vibrio parahemolyticus, and Listeria monocytogenes suggesting that it may be involved in immune response reactions in abalone. Copyright 2010 Elsevier Ltd. All rights reserved.

  9. Isolation and identification of lactic acid bacteria from abalone (Haliotis asinina as a potential candidate of probiotic

    Directory of Open Access Journals (Sweden)

    YAYAN SOFYAN

    2010-01-01

    Full Text Available Sarkono, Faturrahman, Sofyan Y. 2010. Isolation and identification of lactic acid bacteria from abalone (Haliotis asinina as a potential candidate of probiotic. Nusantara Bioscience 2: 38-42. The purpose of this study was to isolate, select and characterize lactic acid bacteria (LAB from abalone as a potential candidate probiotic in abalone cultivation system. Selective isolation of LAB performed using de Man Rogosa Sharpe medium. LAB isolate that potential as probiotics was screened. Selection was based on its ability to suppress the growth of pathogenic bacteria, bacterial resistance to acidic conditions and bacterial resistance to bile salts (bile. Further characterization and identification conducted to determine the species. The results showed that two of the ten isolates potential to be developed as probiotic bacteria that have the ability to inhibit several pathogenic bacteria such as Eschericia coli, Bacillus cereus dan Staphylococus aureus, able to grow at acidic condition and bile tolerance during the incubation for 24 hour. Based on the API test kit, the both of isolate identified as members of the species Lactobacillus paracasei ssp. paracasei.

  10. Sperm competition promotes diversity of sperm bundles in Ohomopterus ground beetles

    Science.gov (United States)

    Takami, Yasuoki; Sota, Teiji

    2007-07-01

    Diversification of sperm morphology has been investigated in the context of sperm competition, but the adaptive significance of sperm bundles is still unclear. In analyzing 10 taxa of the genus Carabus subgenus Ohomopterus and one related Carabus ground beetles, we found that dimorphic sperm bundles occurred in most species with varied degrees of bimodality, whereas sperm were generally monomorphic. Comparative analyses with phylogenetically independent contrasts revealed that the sizes of large and small sperm bundles evolved more rapidly than, and were not correlated with, the length of sperm, suggesting more intense selection on sperm bundle sizes and their independent responses to different evolutionary forces. The size of large sperm bundles was positively correlated with male genital morphology (pertinent to displacement of rival spermatophores) and postcopulatory guarding duration as well as male body length, suggesting that larger sperm bundles have been favored when the risk of spermatophore displacement is high. Larger sperm bundles may be advantageous because of their ability to migrate more rapidly into the spermatheca. In contrast, no clear association was detected between the small sperm bundle size and mating traits despite its rapid diversification. The present study provides the first record of heteromorphic sperm bundles, the diversity of which may be promoted by sperm competition.

  11. Phenotypic engineering of sperm-production rate confirms evolutionary predictions of sperm competition theory.

    Science.gov (United States)

    Sekii, Kiyono; Vizoso, Dita B; Kuales, Georg; De Mulder, Katrien; Ladurner, Peter; Schärer, Lukas

    2013-04-22

    Sperm production is a key male reproductive trait and an important parameter in sperm competition theory. Under sperm competition, paternity success is predicted to depend strongly on male allocation to sperm production. Furthermore, because sperm production is inherently costly, individuals should economize in sperm expenditure, and conditional adjustment of the copulation frequency according to their sperm availability may be expected. However, experimental studies showing effects of sperm production on mating behaviour and paternity success have so far been scarce, mainly because sperm production is difficult to manipulate directly in animals. Here, we used phenotypic engineering to manipulate sperm-production rate, by employing dose-dependent RNA interference (RNAi) of a spermatogenesis-specific gene, macbol1, in the free-living flatworm Macrostomum lignano. We demonstrate (i) that our novel dose-dependent RNAi approach allows us to induce high variability in sperm-production rate; (ii) that a reduced sperm-production rate is associated with a decreased copulation frequency, suggesting conditional adjustment of mating behaviour; and (iii) that both sperm production and copulation frequency are important determinants of paternity success in a competitive situation, as predicted by sperm competition theory. Our study clearly documents the potential of phenotypic engineering via dose-dependent RNAi to test quantitative predictions of evolutionary theory.

  12. Involvement of clip-domain serine protease in the anti-Vibrio immune response of abalone (Haliotis discus hannai)-Molecular cloning, characterization and functional analysis.

    Science.gov (United States)

    Hu, Jian-Jian; Chen, Yu-Lei; Duan, Xue-Kun; Jin, Teng-Chuan; Li, Yue; Zhang, Ling-Jing; Liu, Guang-Ming; Cao, Min-Jie

    2018-01-01

    Vibrio parahemolyticus (V. parahemolyticus) is a major pathogen for abalone, an important economical shellfish in coastal area of China. There is little known about the abalone innate immune system against pathogen infection. Clip-domain serine proteases (cSPs) are increasingly recognized to play important roles in host immune defense in invertebrates. In this study, we cloned a cSP (Hdh-cSP) from abalone (Haliotis discus hannai). We found out that Hdh-cSP was widely expressed in multiple tissues of abalone, with highest level in the immune-like organ, hepatopancreas. V. parahemolyticus infection induced significantly elevated expression of Hdh-cSP in addition to better-characterized innate immune component genes including Rel/NF-κB, allograft inflammatory factor (ALInFa), macrophage expressed protein (MEP) and caspase-8. Importantly, the silencing of Hdh-cSP reduced the expression of these genes, suggesting that Hdh-cSP was an upstream regulatory factor in V. parahemolyticus infection. Further analysis showed that apoptosis of hemocytes was inhibited when the transcription of Hdh-cSP was knocked down, suggesting that Hdh-cSP participated in cell apoptosis by regulation of caspase 8 expression in V. parahemolyticus infection. Therefore, our study established an important role of cSP in the innate immunity against V. parahemolyticus infection in abalone. Copyright © 2017 Elsevier Ltd. All rights reserved.

  13. Elucidating the effects of arginine and lysine on a monoclonal antibody C-terminal lysine variation in CHO cell cultures.

    Science.gov (United States)

    Zhang, Xintao; Tang, Hongping; Sun, Ya-Ting; Liu, Xuping; Tan, Wen-Song; Fan, Li

    2015-08-01

    C-terminal lysine variants are commonly observed in monoclonal antibodies (mAbs) and found sensitive to process conditions, especially specific components in culture medium. The potential roles of media arginine (Arg) and lysine (Lys) in mAb heavy chain C-terminal lysine processing were investigated by monitoring the lysine variant levels under various Arg and Lys concentrations. Both Arg and Lys were found to significantly affect lysine variant level. Specifically, lysine variant level increased from 18.7 to 31.8 % when Arg and Lys concentrations were increased from 2 to 10 mM. Since heterogeneity of C-terminal lysine residues is due to the varying degree of proteolysis by basic carboxypeptidases (Cps), enzyme (basic Cps) level, pH conditions, and product (Arg and Lys) inhibition, which potentially affect the enzymatic reaction, were investigated under various Arg and Lys conditions. Enzyme level and pH conditions were found not to account for the different lysine variant levels, which was evident from the minimal variation in transcription level and intracellular pH. On the other hand, product inhibition effect of Arg and Lys on basic Cps was evident from the notable intracellular and extracellular Arg and Lys concentrations comparable with Ki values (inhibition constant) of basic Cps and further confirmed by cell-free assays. Additionally, a kinetic study of lysine variant level during the cell culture process enabled further characterization of the C-terminal lysine processing.

  14. On the origin of sperm epigenetic heterogeneity.

    Science.gov (United States)

    Laurentino, Sandra; Borgmann, Jennifer; Gromoll, Jörg

    2016-05-01

    The influence of epigenetic modifications on reproduction and on the function of male germ cells has been thoroughly demonstrated. In particular, aberrant DNA methylation levels in sperm have been associated with abnormal sperm parameters, lower fertilization rates and impaired embryo development. Recent reports have indicated that human sperm might be epigenetically heterogeneous and that abnormal DNA methylation levels found in the sperm of infertile men could be due to the presence of sperm populations with different epigenetic quality. However, the origin and the contribution of different germ cell types to this suspected heterogeneity remain unclear. In this review, we focus on sperm epigenetics at the DNA methylation level and its importance in reproduction. We take into account the latest developments and hypotheses concerning the functional significance of epigenetic heterogeneity coming from the field of stem cell and cancer biology and discuss the potential importance and consequences of sperm epigenetic heterogeneity for reproduction, male (in)fertility and assisted reproductive technologies (ART). Based on the current information, we propose a model in which spermatogonial stem cell variability, either intrinsic or due to external factors (such as endocrine action and environmental stimuli), can lead to epigenetic sperm heterogeneity, sperm epimutations and male infertility. The elucidation of the precise causes for epimutations, the conception of adequate therapeutic options and the development of sperm selection technologies based on epigenetic quality should be regarded as crucial to the improvement of ART outcome in the near future. © 2016 Society for Reproduction and Fertility.

  15. Sperm competition and reproductive mode influence sperm dimensions and structure among snakes.

    Science.gov (United States)

    Tourmente, Maximiliano; Gomendio, Montserrat; Roldan, Eduardo R S; Giojalas, Laura C; Chiaraviglio, Margarita

    2009-10-01

    The role of sperm competition in increasing sperm length is a controversial issue, because findings from different taxa seem contradictory. We present a comparative study of 25 species of snakes with different levels of sperm competition to test whether it influences the size and structure of different sperm components. We show that, as levels of sperm competition increase, so does sperm length, and that this elongation is largely explained by increases in midpiece length. In snakes, the midpiece is comparatively large and it contains structures, which in other taxa are present in the rest of the flagellum, suggesting that it may integrate some of its functions. Thus, increases in sperm midpiece size would result in more energy as well as greater propulsion force. Sperm competition also increases the area occupied by the fibrous sheath and outer dense fibers within the sperm midpiece, revealing for the first time an effect upon structural elements within the sperm. Finally, differences in male-male encounter rates between oviparous and viviparous species seem to lead to differences in levels of sperm competition. We conclude that the influence of sperm competition upon different sperm components varies between taxa, because their structure and function is different.

  16. No evidence of trade-offs in the evolution of sperm numbers and sperm size in mammals.

    Science.gov (United States)

    Tourmente, M; Delbarco Trillo, J; Roldan, E R S

    2015-10-01

    Post-copulatory sexual selection, in the form sperm competition, has influenced the evolution of several male reproductive traits. However, theory predicts that sperm competition would lead to trade-offs between numbers and size of spermatozoa because increased costs per cell would result in a reduction of sperm number if both traits share the same energetic budget. Theoretical models have proposed that, in large animals, increased sperm size would have minimal fitness advantage compared with increased sperm numbers. Thus, sperm numbers would evolve more rapidly than sperm size under sperm competition pressure. We tested in mammals whether sperm competition maximizes sperm numbers and size, and whether there is a trade-off between these traits. Our results showed that sperm competition maximizes sperm numbers in eutherian and metatherian mammals. There was no evidence of a trade-off between sperm numbers and sperm size in any of the two mammalian clades as we did not observe any significant relationship between sperm numbers and sperm size once the effect of sperm competition was taken into account. Maximization of both numbers and size in mammals may occur because each trait is crucial at different stages in sperm's life; for example size-determined sperm velocity is a key determinant of fertilization success. In addition, numbers and size may also be influenced by diverse energetic budgets required at different stages of sperm formation. © 2015 European Society For Evolutionary Biology. Journal of Evolutionary Biology © 2015 European Society For Evolutionary Biology.

  17. Temporal trends in sperm count

    DEFF Research Database (Denmark)

    Levine, Hagai; Jørgensen, Niels; Martino-Andrade, Anderson

    2017-01-01

    , method of measuring SC and semen volume, exclusion criteria and indicators of completeness of covariate data]. The slopes of SC and TSC were estimated as functions of sample collection year using both simple linear regression and weighted meta-regression models and the latter were adjusted for pre......-determined covariates and modification by fertility and geographic group. Assumptions were examined using multiple sensitivity analyses and nonlinear models. OUTCOMES: SC declined significantly between 1973 and 2011 (slope in unadjusted simple regression models -0.70 million/ml/year; 95% CI: -0.72 to -0.69; P ...BACKGROUND: Reported declines in sperm counts remain controversial today and recent trends are unknown. A definitive meta-analysis is critical given the predictive value of sperm count for fertility, morbidity and mortality. OBJECTIVE AND RATIONALE: To provide a systematic review and meta-regression...

  18. Nuclear microscopy of sperm cell elemental structure

    Energy Technology Data Exchange (ETDEWEB)

    Bench, G.S.; Balhorn, R.; Friz, A.M.; Freeman, S.P.H.T.

    1994-09-28

    Theories suggest there is a link between protamine concentrations in individual sperm and male fertility. Previously, biochemical analyses have used pooled samples containing millions of sperm to determine protamine concentrations. These methods have not been able to determine what percentage of morphologically normal sperm are biochemically defective and potentially infertile. Nuclear microscopy has been utilized to measure elemental profiles at the single sperm level. By measuring the amount of phosphorus and sulfur, the total DNA and protamine content in individual sperm from fertile bull and mouse semen have been determined. These values agree with results obtained from other biochemical analyses. Nuclear microscopy shows promise for measuring elemental profiles in the chromatin of individual sperm. The technique may be able to resolve theories regarding the importance of protamines to male fertility and identify biochemical defects responsible for certain types of male infertility.

  19. Sperm interaction with the female reproductive tract.

    Science.gov (United States)

    Druart, X

    2012-08-01

    Sperm transit in the female tract is a critical event for the success of fertilization. From their deposition in the vagina to final migration in the oviduct, sperm pass through the different compartments of the genital tract in which they encounter different environments. The cervix and the uterotubal junction (UTJ) are two barriers with different relative importance according to the species. The protein composition, the degree of glycosylation and the hydration of the cervical mucus change during the oestrous cycle. Several sperm surface proteins are associated with their migration through the cervical mucus and the UTJ. Data regarding the interaction of sperm with secretions of the epithelial tissue lining the different compartments of the female genital tract during the sperm transit are reviewed, with a particular emphasis on the migration of sperm through the cervix. © 2012 Blackwell Verlag GmbH.

  20. Enzymatic production of 5-aminovalerate from L-lysine using L-lysine monooxygenase and 5-aminovaleramide amidohydrolase.

    Science.gov (United States)

    Liu, Pan; Zhang, Haiwei; Lv, Min; Hu, Mandong; Li, Zhong; Gao, Chao; Xu, Ping; Ma, Cuiqing

    2014-07-11

    5-Aminovalerate is a potential C5 platform chemical for synthesis of valerolactam, 5-hydroxyvalerate, glutarate, and 1,5-pentanediol. It is a metabolite of l-lysine catabolism through the aminovalerate pathway in Pseudomonas putida. L-Lysine monooxygenase (DavB) and 5-aminovaleramide amidohydrolase (DavA) play key roles in the biotransformation of L-lysine into 5-aminovalerate. Here, DavB and DavA of P. putida KT2440 were expressed, purified, and coupled for the production of 5-aminovalerate from L-lysine. Under optimal conditions, 20.8 g/L 5-aminovalerate was produced from 30 g/L L-lysine in 12 h. Because L-lysine is an industrial fermentation product, the two-enzyme coupled system presents a promising alternative for the production of 5-aminovalerate.

  1. Molecular evolution of the lysine biosynthetic pathways.

    Science.gov (United States)

    Velasco, A M; Leguina, J I; Lazcano, A

    2002-10-01

    Among the different biosynthetic pathways found in extant organisms, lysine biosynthesis is peculiar because it has two different anabolic routes. One is the diaminopimelic acid pathway (DAP), and the other over the a-aminoadipic acid route (AAA). A variant of the AAA route that includes some enzymes involved in arginine and leucine biosyntheses has been recently reported in Thermus thermophilus (Nishida et al. 1999). Here we describe the results of a detailed genomic analysis of each of the sequences involved in the two lysine anabolic routes, as well as of genes from other routes related to them. No evidence was found of an evolutionary relationship between the DAP and AAA enzymes. Our results suggest that the DAP pathway is related to arginine metabolism, since the lysC, asd, dapC, dapE, and lysA genes from lysine biosynthesis are related to the argB, argC, argD, argE, and speAC genes, respectively, whose products catalyze different steps in arginine metabolism. This work supports previous reports on the relationship between AAA gene products and some enzymes involved in leucine biosynthesis and the tricarboxylic acid cycle (Irvin and Bhattacharjee 1998; Miyazaki et al. 2001). Here we discuss the significance of the recent finding that several genes involved in the arginine (Arg) and leucine (Leu) biosynthesis participate in a new alternative route of the AAA pathway (Miyazaki et al. 2001). Our results demonstrate a clear relationship between the DAP and Arg routes, and between the AAA and Leu pathways.

  2. Swedish sperm donors are driven by altruism, but shortage of sperm donors leads to reproductive travelling.

    Science.gov (United States)

    Ekerhovd, Erling; Faurskov, Anders; Werner, Charlotte

    2008-01-01

    Swedish legislation requires that sperm donors are identifiable to offspring. In Denmark sperm donors remain anonymous. The aim of this study was to examine sperm donation in Sweden by identifying socio-demographic backgrounds, motivations and attitudes among donors and to describe options and plans of sperm recipients. Furthermore, the willingness of Swedish health care providers to assist in treatment abroad, where sperm from an anonymous donor were to be used, was assessed. The extent of travelling to Denmark for reproductive purposes was also examined. Thirty Swedish sperm donors completed a questionnaire and were interviewed about their backgrounds, motivations and attitudes. Thirty couples where the infertility workup had shown azoospermia were interviewed about their options for achieving parenthood. The willingness to assist in fertility treatment abroad and the extent of reproductive cross border travelling were assessed by interviewing health care providers and by contacting Danish clinics. Almost all donors were Caucasian. The main motivation for sperm donors was to help others. Owing to shortage of sperm donors many Caucasian recipients intended to have treatment abroad. For most non-Caucasian recipients sperm from a donor of appropriate ethnicity were not available in Sweden. Whether the sperm donor was anonymous or identifiable was not of major importance to most sperm recipients. Health care providers expressed unanimous willingness to assist in treatment with sperm from an anonymous donor. Our inquiry indicated that more than 250 Swedish sperm recipients travel to Denmark annually. Identifiable sperm donors are driven by altruistic motives, but shortage of sperm donors leads to reproductive travelling. Recruitment strategies to increase the number of sperm donors in Sweden are therefore warranted.

  3. Ejaculated mouse sperm enter cumulus-oocyte complexes more efficiently in vitro than epididymal sperm.

    Science.gov (United States)

    Li, Honggang; Hung, Pei-Hsuan; Suarez, Susan S

    2015-01-01

    The mouse is an established and popular animal model for studying reproductive biology. Epididymal mouse sperm, which lack exposure to secretions of male accessory glands and do not precisely represent ejaculated sperm for the study of sperm functions, have been almost exclusively used in studies. We compared ejaculated and epididymal sperm in an in vitro fertilization setting to examine whether ejaculated sperm enter cumulus-oocyte complexes more efficiently. In order to prepare sperm for fertilization, they were incubated under capacitating conditions. At the outset of incubation, ejaculated sperm stuck to the glass surfaces of slides and the incidences of sticking decreased with time; whereas, very few epididymal sperm stuck to glass at any time point, indicating differences in surface charge. At the end of the capacitating incubation, when sperm were added to cumulus-oocyte complexes, the form of flagellar movement differed dramatically; specifically, ejaculated sperm predominantly exhibited increased bending on one side of the flagellum (a process termed pro-hook hyperactivation), while epididymal sperm equally exhibited increased bending on one or the other side of the flagellum (pro-hook or anti-hook hyperactivation). This indicates that accessory sex gland secretions might have modified Ca2+ signaling activities in sperm, because the two forms of hyperactivation are reported to be triggered by different Ca2+ signaling patterns. Lastly, over time, more ejaculated than epididymal sperm entered the cumulus oocyte complexes. We concluded that modification of sperm by male accessory gland secretions affects the behavior of ejaculated sperm, possibly providing them with an advantage over epididymal sperm for reaching the eggs in vivo.

  4. Artificial insemination with donor sperm (AID): heterogeneity in sperm banking facilities in a single country (Belgium)

    Science.gov (United States)

    Thijssen, A.; Dhont, N.; Vandormael, E.; Cox, A.; Klerkx, E.; Creemers, E.; Ombelet, W.

    2014-01-01

    Due to the high inflow of foreign patients seeking cross-border reproductive care in Belgium and the increased number of lesbian couples and single women who call for artificial insemination with donor sperm (AID), Belgian sperm banks nowadays face a shortage in donor sperm. However, since there is no central registration system for sperm donors in Belgium, no figures are currently available supporting this statement. Therefore a study was performed to obtain a detailed overview of the sperm banking facilities in Belgium. Questionnaires were sent to all Belgian centres for assisted reproduction with laboratory facilities (n = 18) to report on their sperm banking methods. The results showed that 82% of the centres rely partially or completely on foreign donor sperm. Moreover, four of the thirteen centres that have their own sperm bank use imported donor sperm in > 95% AID cycles. Our results show that in 63% of the Belgian AID cycles imported Danish donor sperm is being used. Donor recruitment is mainly performed through the centre’s website (61%) or by distributing flyers in the centre (46%) and 9 to 180 potential donors have been recruited per centre in 2013. Eventually, 15 to 50% of these candidate donors were accepted. Different criteria for donor acceptance are handled by the centres: donor age limits range from 18-25 to 36-46 years old, and thresholds for sperm normality differ considerably. We can conclude that a wide variation in methods associated with sperm banking is observed in Belgian centres. PMID:25009728

  5. Intraspecific variation in sperm length is negatively related to sperm competition in passerine birds.

    Science.gov (United States)

    Kleven, Oddmund; Laskemoen, Terje; Fossøy, Frode; Robertson, Raleigh J; Lifjeld, Jan T

    2008-02-01

    Spermatozoa are among the most diversified cells in the animal kingdom, but the underlying evolutionary forces affecting intraspecific variation in sperm morphology are poorly understood. It has been hypothesized that sperm competition is a potent selection pressure on sperm variation within species. Here, we examine intraspecific variation in total sperm length of 22 wild passerine bird species (21 genera, 11 families) in relation to the risk of sperm competition, as expressed by the frequency of extrapair paternity and relative testis size. We demonstrate, by using phylogenetic comparative methods, that between-male variation in sperm length within species is closely and negatively linked to the risk of sperm competition. This relationship was even stronger when only considering species in which data on sperm length and extrapair paternity originated from the same populations. Intramale variation in sperm length within species was also negatively, although nonsignificantly, related to sperm competition risk. Our findings suggest that postcopulatory sexual selection is a powerful evolutionary force reducing the intraspecific phenotypic variation in sperm-size traits, potentially driving the diversification of sperm morphology across populations and species.

  6. Sperm cryopreservation update: Cryodamage, markers, and factors affecting the sperm freezability in pigs.

    Science.gov (United States)

    Yeste, Marc

    2016-01-01

    Cryopreservation is the most efficient method for long-term preservation of mammalian sperm. However, freeze-thawing procedures may strongly impair the sperm function and survival and thus decrease the reproductive performance. In addition, the sperm resilience to withstand cryopreservation, also known as freezability, presents a high individual variability. The present work summarizes the principles of cryoinjury and the relevance of permeating and nonpermeating cryoprotective agents. Descriptions about sperm cryodamage are mainly focused on boar sperm, but reference to other mammalian species is also made when relevant. Main cryoinjuries not only regard to sperm motility and membrane integrity, but also to the degradation effect exerted by freeze-thawing on other important components for sperm fertilizing ability, such as mRNAs. After delving into the main differences between good and poor freezability boar ejaculates, those protein markers predicting the sperm ability to sustain cryopreservation are also mentioned. Moreover, factors that may influence sperm freezability, such as season, diet, breed, or ejaculate fractions are discussed, together with the effects of different additives, like seminal plasma and antioxidants. After briefly referring to the effects of long-term sperm preservation in frozen state and the reproductive performance of frozen-thawed boar sperm, this work speculates with new research horizons on the preservation of boar sperm, such as vitrification and freeze-drying. Copyright © 2016 Elsevier Inc. All rights reserved.

  7. Varicocele Negatively Affects Sperm Mitochondrial Respiration.

    Science.gov (United States)

    Ferramosca, Alessandra; Albani, Denise; Coppola, Lamberto; Zara, Vincenzo

    2015-10-01

    To evaluate the effect of varicocele on oxidative stress, sperm mitochondrial respiratory efficiency, sperm morphology, and semen parameters. A total of 20 patients with varicocele and 20 normozoospermic subjects without varicocele (control group) were recruited from a medical center for reproductive biology. The levels of serum reactive oxygen metabolites and seminal lipid peroxides were assessed for both control and varicocele subjects. Sperm deoxyribonucleic acid fragmentation was measured by sperm chromatin dispersion test. Mitochondrial respiratory activity was evaluated with a polarographic assay of oxygen consumption carried out in hypotonically treated sperm cells. In this study, varicocele patients were compared with men without varicoceles. Oxidative stress was observed in the serum and seminal fluid of varicocele patients. These patients showed an increase of 59% (P <.05) in serum reactive oxygen metabolites and a 3-fold increase in the level of sperm lipid peroxides. A parallel and significant increase (a 2-fold increase; P <.05) in the degree of sperm deoxyribonucleic acid fragmentation was also observed. Varicocele patients showed a 27% decrease (P <.05) in mitochondrial respiratory activity in comparison to the control group. A 32% increase (P <.05) in sperm midpiece defects and a 41% decrease (P <.05) in sperm concentration and motility were also observed. Men with varicocele have increased markers of oxidative stress and decreased mitochondrial respiratory activity. These results correlated with abnormalities in semen parameters. For morphology, these correlated with midpiece defects. Copyright © 2015 Elsevier Inc. All rights reserved.

  8. Hypercholesterolemia Impaired Sperm Functionality in Rabbits

    Science.gov (United States)

    Monclus, Maria A.; Cabrillana, Maria E.; Clementi, Marisa A.; Espínola, Leandro S.; Cid Barría, Jose L.; Vincenti, Amanda E.; Santi, Analia G.; Fornés, Miguel W.

    2010-01-01

    Hypercholesterolemia represents a high risk factor for frequent diseases and it has also been associated with poor semen quality that may lead to male infertility. The aim of this study was to analyze semen and sperm function in diet-induced hypercholesterolemic rabbits. Twelve adult White New Zealand male rabbits were fed ad libitum a control diet or a diet supplemented with 0.05% cholesterol. Rabbits under cholesterol-enriched diet significantly increased total cholesterol level in the serum. Semen examination revealed a significant reduction in semen volume and sperm motility in hypercholesterolemic rabbits (HCR). Sperm cell morphology was seriously affected, displaying primarily a “folded head”-head fold along the major axe-, and the presence of cytoplasmic droplet on sperm flagellum. Cholesterol was particularly increased in acrosomal region when detected by filipin probe. The rise in cholesterol concentration in sperm cells was determined quantitatively by Gas chromatographic-mass spectrometric analyses. We also found a reduction of protein tyrosine phosphorylation in sperm incubated under capacitating conditions from HCR. Interestingly, the addition of Protein Kinase A pathway activators -dibutyryl-cyclic AMP and iso-butylmethylxanthine- to the medium restored sperm capacitation. Finally, it was also reported a significant decrease in the percentage of reacted sperm in the presence of progesterone. In conclusion, our data showed that diet-induced hypercholesterolemia adversely affects semen quality and sperm motility, capacitation and acrosomal reaction in rabbits; probably due to an increase in cellular cholesterol content that alters membrane related events. PMID:20976152

  9. Dietary ascorbic acid modulates the expression profile of stress protein genes in hepatopancreas of adult Pacific abalone Haliotis discus hannai Ino.

    Science.gov (United States)

    Wu, Chenglong; Wang, Jia; Xu, Wei; Zhang, Wenbing; Mai, Kangsen

    2014-12-01

    This study was conducted to investigate the effects of dietary ascorbic acid (AA) on transcriptional expression patterns of antioxidant proteins, heat shock proteins (HSP) and nuclear factor kappa B (NF-κB) in the hepatopancreas of Pacific abalone Haliotis discus hannai Ino (initial average length: 84.36 ± 0.24 mm) using real-time quantitative PCR assays. L-ascorbyl-2-molyphosphate (LAMP) was added to the basal diet to formulate four experimental diets containing 0.0, 70.3, 829.8 and 4967.5 mg AA equivalent kg(-1) diets, respectively. Each diet was fed to triplicate groups of adult abalone in acrylic tanks (200 L) in a flow-through seawater system. Each tank was stocked with 15 abalone. Animals were fed once daily (17:00) to apparent satiation for 24 weeks. The results showed that the dietary AA (70.3 mg kg(-1)) could significantly up-regulate the expression levels of Cu/Zn superoxide dismutase (SOD), catalase (CAT), glutathione-S-transferase (GST), feritin (FT) and heat shock protein 26 (HSP26) in the hepatopancreas of abalone in this treatment compared to the controls. However, the expression levels of Mn-SOD, glutathione peroxidase (GPX), thioredoxin peroxidase (TPx), selenium-binding protein (SEBP), HSP70 and HSP90 were significantly down-regulated. Compared with those in the group with 70.3 mg kg(-1) dietary AA, the expression levels of CAT, GST and HSP26 were decreased in abalone fed with very high dietary AA (4967.5 mg kg(-1)). In addition, significant up-regulations of expression levels of Mn-SOD, GPX, TPx, SEBP, FT, HSP70, HSP90 and NF-κB were observed in abalone fed with apparently excessive dietary AA (829.8 and 4967.5 mg kg(-1)) as compared to those fed 70.3 mg kg(-1) dietary AA. These findings showed that dietary AA influenced the expression levels of antioxidant proteins, heat shock proteins and NF-κB in the hepatopancreas of abalone at transcriptional level. Levels of dietary AA that appeared adequate (70.3 mg kg(-1)) reduced the oxidative stress

  10. Sperm competition selects beyond relative testes size in birds

    OpenAIRE

    Lüpold S; Linz GM; Rivers JW; Westneat DF; Birkhead TR

    2009-01-01

    Sperm morphology varies considerably across taxa and postcopulatory sexual selection is thought to be one of the main forces responsible for this diversity. Several studies have investigated the effects of the variation in sperm design on sperm function but the consequences of variation in sperm design on testis morphology have been overlooked. Testes size or architecture may determine the size of the sperm they produce and selection for longer sperm may require concomitant adaptations in the...

  11. Current status and potential of morphometric sperm analysis

    OpenAIRE

    Maroto-Morales, Alejandro; García-Álvarez, Olga; Ramón, Manuel; Martínez-Pastor, Felipe; Fernández-Santos, M. Rocío; Soler, Ana J.; Garde, José Julián

    2016-01-01

    The spermatozoon is the most diverse cell type known and this diversity is considered to reflect differences in sperm function. How the diversity in sperm morphology arose during speciation and what role the different specializations play in sperm function, however, remain incompletely characterized. This work reviews the hypotheses proposed to explain sperm morphological evolution, with a focus on some aspects of sperm morphometric evaluation; the ability of morphometrics to predict sperm cr...

  12. Single blastocyst transfer after ICSI from ejaculate spermatozoa, percutaneous epididymal sperm aspiration (PESA) or testicular sperm extraction (TESE)

    OpenAIRE

    Nilsson, Staffan; Waldenström, Urban; Engström, Ann-Britt; Hellberg, Dan

    2007-01-01

    Purpose: To investigate the outcome of IVF following intracytoplasmic sperm injection (ICSI) from ejaculate, percutaneous epididymal sperm aspiration (PESA) and testicular sperm extraction (TESE), with subsequent blastocyst culture and single blastocyst transfer.

  13. Biomarkers of arginine and lysine excess.

    Science.gov (United States)

    Luiking, Yvette C; Deutz, Nicolaas E P

    2007-06-01

    Arginine supplementation is used in several disease states. In arginine-deficient states, supplementation is a logical choice of therapy. However, the definition of an arginine-deficient state is complex. For example, plasma arginine levels could be within normal range but intracellular arginine levels could be reduced because of membrane transport problems. Lysine competes with arginine for transport into the cell. In these situations, arginine supplementation of higher than required levels is proposed. Arginine has several important functions in metabolism as it is a precursor of metabolically active components such as nitric oxide (NO), ornithine, creatine, and polyamines. Supplementing arginine in excess could potentially overstimulate metabolism via enhanced production of NO. NO is a reactive component that, via production of radicals, will inactivate proteins. NO is also a powerful vasodilator, which could lead to severe hemodynamic instability. A good marker for excess supplementation of arginine or lysine could be an increased or reduced production rate of NO. However, NO production is difficult to measure because NO is a very labile component and is rapidly oxidized in blood. Stable isotope-labeled arginine and citrulline are used to trace the arginine-NO route. During supplementation of arginine in septic pigs or patients in septic shock, NO production, measured with stable isotope technology, is enhanced.

  14. Structural characterization and osteogenic bioactivity of a sulfated polysaccharide from pacific abalone (Haliotis discus hannai Ino).

    Science.gov (United States)

    Song, Shuang; Zhang, Bao; Wu, Sufeng; Huang, Lu; Ai, Chunqing; Pan, Jinfeng; Su, Yi-Cheng; Wang, Zhongfu; Wen, Chengrong

    2018-02-15

    Bone morphogenic protein-2 (BMP-2) is known to promote osteogenesis. To find novel adjuvants to enhance the activity of BMP-2, the present study investigated the structure BMP-2-induced osteogenic activity of a water-soluble polysaccharide from the gonad of pacific abalone (Haliotis discus hannai Ino) named AGSP. Through analysis of aldobiouronic acids released from AGSP, monosaccharide composition comparison of AGSP and its reduced product, and methylation analysis and NMR analysis of AGSP and its desulfated derivative, the main structure residue of AGSP was determined as →3)-GlcA(1→3)-Gal(1→ with sulfated branches comprised of prevelant Gal and minor Glc, and →4)-β-GlcA(1→2)-α-Man(1→ residue was also found. AGSP possessed a sulfate content of 12.4% with a relative molecular weight of 6.6kDa. AGSP strengthened alkaline phosphatase activity induced by BMP-2 in a dose dependent manner at 10-200μg/mL with 425% enhancement being observed at 200μg/mL, indicating AGSP could be an adjuvant candidate to enhance osteogenic activity of BMP-2. Copyright © 2017 Elsevier Ltd. All rights reserved.

  15. Effects of polysaccharides from abalone viscera (Haliotis discus hannai Ino) on MGC 803 cells proliferation.

    Science.gov (United States)

    Zhang, Rui-Juan; Shi, Yan; Zheng, Jing; Mao, Xiao-Mei; Liu, Zhi-Yu; Chen, Qing-Xi; Wang, Qin

    2018-01-01

    The polysaccharides (AVP) was obtained from abalone (Haliotis discus hannai Ino) viscera, using the alkaline protease to enzymolysis, sevage method and repeated freezing and thawing method to remove protein and hydrogen peroxide method to depigment. The total sugar content was 46.27±1.5% and uronic acid, sulfate radical, hexosamine and protein contents were 17.44±0.22%, 16.98±0.15%, 0.65±0.02% and 1.64±0.13% in AVP respectively. The main monosaccharide compositions of AVP were d-galactose, d-xylose, d-mannose, d-glucose and d-glucuronic acid. MTT assay showed AVP had a significant anti-tumor activity to gastric carcinoma cells, especially to MGC 803, while it had no influence upon proliferation of normal stomach cells GES 1. The results of Morphological changes, cell migration ability and AO/EB staining indicated that MGC803 cells underwent apoptosis in a dose-dependent manner induced by AVP. Moreover, the western blotting results showed that the expressions of survivin, Bcl-2 and VEGF were decreased, while the expression of Bax and p53 were increased in a dose-dependent manner of AVP. The results suggested that AVP might be a potential anti-tumor agent securely and naturally. Copyright © 2017 Elsevier B.V. All rights reserved.

  16. Characterization of defensin gene from abalone Haliotis discus hannai and its deduced protein

    Science.gov (United States)

    Hong, Xuguang; Sun, Xiuqin; Zheng, Minggang; Qu, Lingyun; Zan, Jindong; Zhang, Jinxing

    2008-11-01

    Defensin is one of preserved ancient host defensive materials formed in biological evolution. As a regulator and effector molecule, it is very important in animals’ acquired immune system. This paper reports the defensin gene from the mixed liver and kidney cDNA library of abalone Haliotis discus hannai Ino. Sequence analysis shows that the gene sequence of full-length cDNA encodes 42 mature peptides (including six Cys), molecular weight of 4 323 Da, and pI of 8.02. Amino acid sequence homology analysis shows that the peptides are highly similar (70% in common) to other insects defensin. Because of a typical insect-defensin structural character of mature peptide in the secondary structure, the polypeptide named Haliotis discus defensin (hd-def), a novel of antimicrobial peptides, belongs to insects defensin subfamily. The RT-PCR result of Haliotis discus defensin shows that the gene can be expressed only in the hepatopancreas by Gram-negative and positive bacteria stimulation, which is ascribed to inducible expression. Therefore, it is revealed that the Haliotis discus defensin gene expression was related to the antibacterial infection of Haliotis discus hannai Ino.

  17. Underwater adhesion of abalone: The role of van der Waals and capillary forces

    International Nuclear Information System (INIS)

    Lin, A.Y.M.; Brunner, R.; Chen, P.Y.; Talke, F.E.; Meyers, M.A.

    2009-01-01

    The observation of the pedal foot of the red abalone Haliotis rufescens reveals the presence of micrometer-scaled setae terminating in nanometer-sized cylindrical fibrils, with some resemblance to those found on the gecko foot. Atomic force microscopy (AFM) pull-off force measurements on a single seta are compared with theoretical estimates for van der Waals attraction obtained through the Johnson-Kendall-Roberts (JKR) equation, approximately 600 nN, and show agreement. The use of the JKR equation is justified through an analysis of the shape of the fibril extremities (parabolic) as well as their diameter (∼200 nm). Measurements under varying humidity conditions indicate that additional capillary interactions play a role, since the pull-off force increases with humidity. It is proposed that both van der Waals and capillary forces play a role in the attachment mechanism of H. rufescens, effectively enabling suction to reach its theoretical limit. Bulk pull-off force measurements on entire live animals yield an average detachment stress of 115 kPa, consistent with theoretical estimates. The setae and nanoscale fibril terminations enable compliance to surfaces with a variety of roughnesses, effectively sealing the interface, in addition to providing capillary and van der Waals forces.

  18. Theoretical characterization of a model of aragonite crystal orientation in red abalone nacre

    International Nuclear Information System (INIS)

    Coppersmith, S N; Gilbert, P U P A; Metzler, R A

    2009-01-01

    Nacre, commonly known as mother-of-pearl, is a remarkable biomineral that in red abalone consists of layers of 400 nm thick aragonite crystalline tablets confined by organic matrix sheets, with the [0 0 1] crystal axes of the aragonite tablets oriented to within ±12 deg. from the normal to the layer planes. Recent experiments demonstrate that greater orientational order develops over a distance of tens of layers from the prismatic boundary at which nacre formation begins. Our previous simulations of a model in which the order develops because of differential tablet growth rates (oriented tablets growing faster than misoriented ones) yield patterns of tablets that agree qualitatively and quantitatively with the experimental measurements. This paper presents an analytical treatment of this model, focusing on how the dynamical development and eventual degree of order depend on model parameters. Dynamical equations for the probability distributions governing tablet orientations are introduced whose form can be determined from symmetry considerations and for which substantial analytic progress can be made. Numerical simulations are performed to relate the parameters used in the analytic theory to those in the microscopic growth model. The analytic theory demonstrates that the dynamical mechanism is able to achieve a much higher degree of order than naive estimates would indicate

  19. Current status and potential of morphometric sperm analysis

    Directory of Open Access Journals (Sweden)

    Alejandro Maroto-Morales

    2016-01-01

    Full Text Available The spermatozoon is the most diverse cell type known and this diversity is considered to reflect differences in sperm function. How the diversity in sperm morphology arose during speciation and what role the different specializations play in sperm function, however, remain incompletely characterized. This work reviews the hypotheses proposed to explain sperm morphological evolution, with a focus on some aspects of sperm morphometric evaluation; the ability of morphometrics to predict sperm cryoresistance and male fertility is also discussed. For this, the evaluation of patterns of change of sperm head morphometry throughout a process, instead of the study of the morphometric characteristics of the sperm head at different stages, allows a better identification of the males with different sperm cryoconservation ability. These new approaches, together with more studies employing a greater number of individuals, are needed to obtain novel results concerning the role of sperm morphometry on sperm function. Future studies should aim at understanding the causes of sperm design diversity and the mechanisms that generate them, giving increased attention to other sperm structures besides the sperm head. The implementation of scientific and technological advances could benefit the simultaneous examination of sperm phenotype and sperm function, demonstrating that sperm morphometry could be a useful tool for sperm assessment.

  20. Current status and potential of morphometric sperm analysis.

    Science.gov (United States)

    Maroto-Morales, Alejandro; García-Álvarez, Olga; Ramón, Manuel; Martínez-Pastor, Felipe; Fernández-Santos, M Rocío; Soler, A Josefa; Garde, José Julián

    2016-01-01

    The spermatozoon is the most diverse cell type known and this diversity is considered to reflect differences in sperm function. How the diversity in sperm morphology arose during speciation and what role the different specializations play in sperm function, however, remain incompletely characterized. This work reviews the hypotheses proposed to explain sperm morphological evolution, with a focus on some aspects of sperm morphometric evaluation; the ability of morphometrics to predict sperm cryoresistance and male fertility is also discussed. For this, the evaluation of patterns of change of sperm head morphometry throughout a process, instead of the study of the morphometric characteristics of the sperm head at different stages, allows a better identification of the males with different sperm cryoconservation ability. These new approaches, together with more studies employing a greater number of individuals, are needed to obtain novel results concerning the role of sperm morphometry on sperm function. Future studies should aim at understanding the causes of sperm design diversity and the mechanisms that generate them, giving increased attention to other sperm structures besides the sperm head. The implementation of scientific and technological advances could benefit the simultaneous examination of sperm phenotype and sperm function, demonstrating that sperm morphometry could be a useful tool for sperm assessment.

  1. The growing landscape of lysine acetylation links metabolism and cell signalling

    DEFF Research Database (Denmark)

    Choudhary, Chuna Ram; Weinert, Brian Tate; Nishida, Yuya

    2014-01-01

    Lysine acetylation is a conserved protein post-translational modification that links acetyl-coenzyme A metabolism and cellular signalling. Recent advances in the identification and quantification of lysine acetylation by mass spectrometry have increased our understanding of lysine acetylation...

  2. Bioavailability of lysine in heat-treated foods and feedstuffs

    NARCIS (Netherlands)

    McArtney Rutherfurd, S.

    2010-01-01

    During the processing of foodstuffs, lysine can react with other compounds present to form nutritionally unavailable derivatives, the most common example of which are Maillard products. Maillard products can cause serious problems when determining the available lysine content of processed foods or

  3. Effects of infused methionine, lysine and rumen-protected ...

    African Journals Online (AJOL)

    Keratin contains about l}Vo arginine, thus a lysine-induced arginine deficiency may depress fibre production as in the study of Sahlu & Fernandez (1992) with. Angora goats. Supply of both methionine and lysine appeared to limit wool growth of sheep limit fed high roughage diets contain- ing non-protein nitrogen as the ...

  4. Threonine and lysine requirements for maintenance in chickens ...

    African Journals Online (AJOL)

    The maintenance requirement for threonine and lysine were estimated in two different experiments by measuring the nitrogen balance of adult male cockerels. Measured amounts of a diet first-limiting in threonine or lysine were fed by intubation each day for 4 d to give a range of intakes (unbalanced series) of from 0 to 239 ...

  5. Antibiotic and surfactant effects on lysine accumulation by Bacillus ...

    African Journals Online (AJOL)

    The effects of antibiotics and surfactants on lysine accumulation in the culture broth of three strains of Bacillus megaterium (B. megaterium SP 86, B. megaterium SP 76 and B. megaterium SP 14) were investigated. Lincomycin, neomycin and tetracycline stimulated lysine increase in B. megaterium SP 76 and B. megaterium ...

  6. effects of dietary chromium tripicolinate and lysine on growth

    African Journals Online (AJOL)

    AISA

    These results show that CrPic has minimal effects on growth efficiency, while lysine affects significantly growth performance, carcass characteristics and most of plasma metabolites in growing-finishing pigs. Key-words : Pig, chromium, lysine, growth, metabolites, USA. RESUME. EFFETS DU TRIPICOLINATE DE CHROME ...

  7. Digestible lysine levels in diets supplemented with ractopamine

    Directory of Open Access Journals (Sweden)

    Evelar de Oliveira Souza

    2011-10-01

    Full Text Available In order evaluate digestible lysine levels in diets supplemented with 20 ppm of ractopamine on the performance and carcass traits, 64 barrows with high genetic potential at finishing phase were allotted in a completely randomized block design with four digestible lysine levels (0.80, 0.90, 1.00, and 1.10%, eight replicates and two pigs per experimental unit. Initial body weight and pigs' kinship were used as criteria in the blocks formation. Diets were mainly composed of corn and soybean meal supplemented with minerals, vitamins and amino acids to meet pigs' nutritional requirements at the finishing phase, except for digestible lysine. No effect of digestible lysine levels was observed in animal performance. The digestible lysine intake increased linearly by increasing the levels of digestible lysine in the diets. Carcass traits were not influenced by the dietary levels of digestible lysine. The level of 0.80% of digestible lysine in diets supplemented with 20 ppm ractopamine meets the nutritional requirements of castrated male pigs during the finishing phase.

  8. Antibiotic and surfactant effects on lysine accumulation by Bacillus ...

    African Journals Online (AJOL)

    STORAGESEVER

    2008-05-16

    May 16, 2008 ... The effects of antibiotics and surfactants on lysine accumulation in the culture broth of three strains of. Bacillus megaterium (B. megaterium SP 86, B. megaterium SP 76 and B. megaterium SP 14) were investigated. Lincomycin, neomycin and tetracycline stimulated lysine increase in B. megaterium SP 76.

  9. Analysis of Grain Protein, Tryptophan and Lysine Contents of Quality ...

    African Journals Online (AJOL)

    Maize proteins, however, have poor nutritional value for humans, because of reduced content of essential amino acids such as lysine, tryptophan and threonine. Maize proteins contain on an average about 2% lysine, which is less than one-half of the concentration recommended for human nutrition. Therefore, healthy diets ...

  10. A cysteine protease (cathepsin Z) from disk abalone, Haliotis discus discus: Genomic characterization and transcriptional profiling during bacterial infections.

    Science.gov (United States)

    Godahewa, G I; Perera, N C N; Lee, Sukkyoung; Kim, Myoung-Jin; Lee, Jehee

    2017-09-05

    Cathepsin Z (CTSZ) is lysosomal cysteine protease of the papain superfamily. It participates in the host immune defense via phagocytosis, signal transduction, cell-cell communication, proliferation, and migration of immune cells such as monocytes, macrophages, and dendritic cells. Hence, CTSZ is also acknowledged as an acute-phase protein in host immunity. In this study, we sought to identify the CTSZ homolog from disk abalone (AbCTSZ) and characterize it at the molecular, genomic, and transcriptional levels. AbCTSZ encodes a protein with 318 amino acids and a molecular mass of 36kDa. The structure of AbCTSZ reveals amino acid sequences that are characteristic of the signal sequence, pro-peptide, peptidase-C1 papain family cysteine protease domain, mini-loop, HIP motif, N-linked glycosylation sites, active sites, and conserved Cys residues. A pairwise comparison revealed that AbCTSZ shared the highest amino acid homology with its molluscan counterpart from Crassostrea gigas. A multiple alignment analysis revealed the conservation of functionally crucial elements of AbCTSZ, and a phylogenetic study further confirmed a proximal evolutionary relationship with its invertebrate counterparts. Further, an analysis of AbCTSZ genomic structure revealed seven exons separated by six introns, which differs from that of its vertebrate counterparts. Quantitative real time PCR (qPCR) detected the transcripts of AbCTSZ in early developmental stages and in eight different tissues. Higher levels of AbCTSZ transcripts were found in trochophore, gill, and hemocytes, highlighting its importance in the early development and immunity of disk abalone. In addition, we found that viable bacteria (Vibrio parahaemolyticus and Listeria monocytogenes) and bacterial lipopolysaccharides significantly modulated AbCTSZ transcription. Collectively, these lines of evidences suggest that AbCTSZ plays an indispensable role in the innate immunity of disk abalone. Copyright © 2017. Published by Elsevier

  11. Cathepsin L is an immune-related protein in Pacific abalone (Haliotis discus hannai)--Purification and characterization.

    Science.gov (United States)

    Shen, Jian-Dong; Cai, Qiu-Feng; Yan, Long-Jie; Du, Cui-Hong; Liu, Guang-Ming; Su, Wen-Jin; Ke, Caihuan; Cao, Min-Jie

    2015-12-01

    Cathepsin L, an immune-related protein, was purified from the hepatopancreas of Pacific abalone (Haliotis discus hannai) by ammonium sulfate precipitation and column chromatographies of SP-Sepharose and Sephacryl S-200 HR. Purified cathepsin L appeared as two bands with molecular masses of 28.0 and 28.5 kDa (namely cathepsin La and Lb) on SDS-PAGE under reducing conditions, suggesting that it is a glycoprotein. Peptide mass fingerprinting (PMF) analysis revealed that peptide fragments of 95 amino acid residues was high similarity to cathepsin L of pearl oyster (Pinctada fucata). The optimal temperature and pH of cathepsin L were 35 °C and pH 5.5. Cathepsin L was particularly inhibited by cysteine proteinase inhibitors of E-64 and leupeptin, while it was activated by metalloproteinase inhibitors EDTA and EGTA. The full-length cathepsin L cDNA was further cloned from the hepatopancreas by rapid PCR amplification of cDNA ends (RACE). The open reading frame of the enzyme was 981 bp, encoding 327 amino acid residues, with a conserved catalytic triad (Cys134, His273 and Asn293), a potential N-glycosylation site and conserved ERFNIN, GNYD, and GCGG motifs, which are characteristics of cathepsin L. Western blot and proteinase activity analysis revealed that the expression and enzyme activity of cathepsin L were significantly up-regulated in hepatopancreas at 8 h following Vibrio parahaemolyticus infection, demonstrating that cathepsin L is involved in the innate immune system of abalone. Our present study for the first time reported the purification, characterization, molecular cloning, and tissue expression of cathepsin L in abalone. Copyright © 2015 Elsevier Ltd. All rights reserved.

  12. Gonadal Transcriptome Analysis of Pacific Abalone Haliotis discus discus: Identification of Genes Involved in Germ Cell Development.

    Science.gov (United States)

    Yu, Lingyun; Xu, Dongdong; Ye, Huan; Yue, Huamei; Ooka, Shioh; Kondo, Hidehiro; Yazawa, Ryosuke; Takeuchi, Yutaka

    2018-04-03

    Little is known about the molecular mechanisms governing gonadal developmental processes in abalones. Here, we conducted transcriptome analysis of Pacific abalone Haliotis discus discus for gene discovery in the brain, ovary, testis, and unfertilized eggs. Among the annotated unigenes, 48.6% of unigenes were identified by Venn diagram analysis as having universal or tissue-specific expression. Twenty-three genes with gonad-biased gene ontology (GO) terms were first obtained. Secondly, 36 genes were found by screening known gene names related to germ cell development. Finally, 17 genes were obtained by querying the annotated unigene database for zygotically expressed gonadal genes (ovary and testis) and maternally expressed gonadal genes (ovary, testis, and unfertilized eggs) using keywords related to reproduction. To further verify tissue distribution pattern and subcellular localization of these genes, RT-PCR and in situ hybridization were performed using a unigene encoding a germ cell marker, vasa, as control. The results showed that vasa was expressed mainly in the early developmental stages of germ cells in both sexes. One of the candidate genes, vitelline envelope zona pellucida domain protein 12 (ZP12), was expressed in the primordial germ cells of immature gonad and early developmental stages of germ cells of the adult female. The results obtained from the present study suggest that vasa and ZP12 are involved in germ cell development of Pacific abalone and that ZP12 is an especially useful germ cell-specific marker in immature adults. The current gonadal transcriptome profile is an extensive resource for future reproductive molecular biology studies of this species.

  13. Observation of sperm-head vacuoles and sperm morphology under light microscope.

    Science.gov (United States)

    Park, Yong-Seog; Park, Sol; Ko, Duck Sung; Park, Dong Wook; Seo, Ju Tae; Yang, Kwang Moon

    2014-09-01

    The presence of sperm-head vacuoles has been suspected to be deleterious to the outcomes of assisted reproductive technology (ART). It is difficult to accurately distinguish morphologically abnormal sperm with vacuoles under a light microscope. This study was performed to analyze the result of the observation of sperm-head vacuoles using Papanicolaou staining under a light microscope and whether the male partner's age affects these vacuoles. Sperm morphology with vacuoles was evaluated using Papanicolaou staining and observed under a light microscope (400×) in 980 men. The normal morphology was divided into three categories (group A, 14% of normal morphology). The criteria for the sperm-head vacuoles were those given in the World Health Organization manual. For the analysis of the age factor, the participants were divided into the following groups: 26-30 years, 31-35 years, 36-40 years, 41-45 years, and 46-50 years. The percentage of sperm-head vacuoles increased with normal sperm morphology (group A vs. groups B, C) (p<0.05). In the case of the age factor, a statistically significant difference was not observed across any of the age groups. A majority of the sperm-head vacuoles showed a statistically significant difference among normal morphology groups. Therefore, we should consider the probability of the percentage of sperm-head vacuoles not increasing with age but with abnormal sperm morphology. A further study is required to clarify the effect of the sperm-head vacuoles on ART outcomes.

  14. Mitochondrial respiratory efficiency is positively correlated with human sperm motility.

    Science.gov (United States)

    Ferramosca, Alessandra; Provenzano, Sara Pinto; Coppola, Lamberto; Zara, Vincenzo

    2012-04-01

    To correlate sperm mitochondrial respiratory efficiency with variations in sperm motility and with sperm morphologic anomalies. Sperm mitochondrial respiratory activity was evaluated with a polarographic assay of oxygen consumption carried out in hypotonically-treated sperm cells. A possible relationship among sperm mitochondrial respiratory efficiency, sperm motility, and morphologic anomalies was investigated. Mitochondrial respiratory efficiency was positively correlated with sperm motility and negatively correlated with the percentage of immotile spermatozoa. Moreover, midpiece defects impaired mitochondrial functionality. Our data indicate that an increase in sperm motility requires a parallel increase in mitochondrial respiratory capacity, thereby supporting the fundamental role played by mitochondrial oxidative phosphorylation in sperm motility of normozoospermic subjects. These results are of physiopathological relevance because they suggest that disturbances of sperm mitochondrial function and of energy production could be responsible for asthenozoospermia. Copyright © 2012 Elsevier Inc. All rights reserved.

  15. Sperm motility of externally fertilizing fish and amphibians.

    Science.gov (United States)

    Browne, R K; Kaurova, S A; Uteshev, V K; Shishova, N V; McGinnity, D; Figiel, C R; Mansour, N; Agney, D; Wu, M; Gakhova, E N; Dzyuba, B; Cosson, J

    2015-01-01

    We review the phylogeny, sperm competition, morphology, physiology, and fertilization environments of the sperm of externally fertilizing fish and amphibians. Increased sperm competition in both fish and anurans generally increases sperm numbers, sperm length, and energy reserves. The difference between the internal osmolarity and iconicity of sperm cells and those of the aquatic medium control the activation, longevity, and velocity of sperm motility. Hypo-osmolarity of the aquatic medium activates the motility of freshwater fish and amphibian sperm and hyperosmolarity activates the motility of marine fish sperm. The average longevity of the motility of marine fish sperm (~550 seconds) was significantly (P amphibian sperm in general and anurans reversion from internal to external fertilization. Our findings provide a greater understanding of the reproductive biology of externally fertilizing fish and amphibians, and a biological foundation for the further development of reproduction technologies for their sustainable management.

  16. Mitochondrial PKA mediates sperm motility.

    Science.gov (United States)

    Mizrahi, Rashel; Breitbart, Haim

    2014-12-01

    Mitochondria are the major source of ATP to power sperm motility. Phosphorylation of mitochondrial proteins has been proposed as a major regulatory mechanism for mitochondrial bioenergetics. Sperm motility was measured by a computer-assisted analyzer, protein detection by western blotting, membrane potential by tetramethylrhodamine, cellular ATP by luciferase assay and localization of PKA by immuno-electron microscopy. Bicarbonate is essential for the creation of mitochondrial electro-chemical gradient, ATP synthesis and sperm motility. Bicarbonate stimulates PKA-dependent phosphorylation of two 60kDa proteins identified as Tektin and glucose-6-phosphate isomerase. This phosphorylation was inhibited by respiration inhibition and phosphorylation could be restored by glucose in the presence of bicarbonate. However, this effect of glucose cannot be seen when the mitochondrial ATP/ADP exchanger was inhibited indicating that glycolytic-produced ATP is transported into the mitochondria and allows PKA-dependent protein phosphorylation inside the mitochondria. Bicarbonate activates mitochondrial soluble adenylyl cyclase (sAC) which catalyzes cAMP production leading to the activation of mitochondrial PKA. Glucose can overcome the lack of ATP in the absence of bicarbonate but it cannot affect the mitochondrial sAC/PKA system, therefore the PKA-dependent phosphorylation of the 60kDa proteins does not occur in the absence of bicarbonate. Production of CO2 in Krebs cycle, which is converted to bicarbonate is essential for sAC/PKA activation leading to mitochondrial membrane potential creation and ATP synthesis. Copyright © 2014 Elsevier B.V. All rights reserved.

  17. Children conceived after intracytoplasmic sperm injection (ICSI)

    DEFF Research Database (Denmark)

    Mau, C; Juul, A; Main, K M

    2004-01-01

    The aim of the study was to evaluate current medical knowledge about children born after intracytoplasmic sperm injection (ICSI) with respect to congenital malformations, chromosome abnormalities and postnatal growth.......The aim of the study was to evaluate current medical knowledge about children born after intracytoplasmic sperm injection (ICSI) with respect to congenital malformations, chromosome abnormalities and postnatal growth....

  18. Short communication Relationship between sperm plasma ...

    African Journals Online (AJOL)

    Matshidiso MB. Masenya

    2017-01-04

    Jan 4, 2017 ... Abstract. Sperm quality plays an important role in determining fertility. The aim of the study was to examine the relationship between sperm plasma membrane integrity and morphology, and fertility following artificial insemination (AI). A total of 16 ejaculates were collected from three Large White boars using ...

  19. Relationship between sperm plasma membrane integrity and ...

    African Journals Online (AJOL)

    Sperm quality plays an important role in determining fertility. The aim of the study was to examine the relationship between sperm plasma membrane integrity and morphology, and fertility following artificial insemination (AI). A total of 16 ejaculates were collected from three Large White boars using the gloved hand ...

  20. Mass-specific metabolic rate and sperm competition determine sperm size in marsupial mammals.

    Directory of Open Access Journals (Sweden)

    Maximiliano Tourmente

    Full Text Available Two complementary hypotheses have been proposed to explain variation in sperm size. The first proposes that post-copulatory sexual selection favors an increase in sperm size because it enhances sperm swimming speed, which is an important determinant of fertilization success in competitive contexts. The second hypothesis proposes that mass-specific metabolic rate acts as a constraint, because large animals with low mass-specific metabolic rates will not be able to process resources at the rates needed to produce large sperm. This constraint is expected to be particularly pronounced among mammals, given that this group contains some of the largest species on Earth. We tested these hypotheses among marsupials, a group in which mass-specific metabolic rates are roughly 30% lower than those of eutherian mammals of similar size, leading to the expectation that metabolic rate should be a major constraint. Our findings support both hypotheses because levels of sperm competition are associated with increases in sperm size, but low mass-specific metabolic rate constrains sperm size among large species. We also found that the relationship between sperm size and mass-specific metabolic rate is steeper among marsupials and shallower among eutherian mammals. This finding has two implications: marsupials respond to changes in mass-specific metabolic rate by modifying sperm length to a greater extent, suggesting that they are more constrained by metabolic rate. In addition, for any given mass-specific metabolic rate, marsupials produce longer sperm. We suggest that this is the consequence of marsupials diverting resources away from sperm numbers and into sperm size, due to their efficient sperm transport along the female tract and the existence of mechanisms to protect sperm.

  1. Mass-specific metabolic rate and sperm competition determine sperm size in marsupial mammals.

    Science.gov (United States)

    Tourmente, Maximiliano; Gomendio, Montserrat; Roldan, Eduardo R S

    2011-01-01

    Two complementary hypotheses have been proposed to explain variation in sperm size. The first proposes that post-copulatory sexual selection favors an increase in sperm size because it enhances sperm swimming speed, which is an important determinant of fertilization success in competitive contexts. The second hypothesis proposes that mass-specific metabolic rate acts as a constraint, because large animals with low mass-specific metabolic rates will not be able to process resources at the rates needed to produce large sperm. This constraint is expected to be particularly pronounced among mammals, given that this group contains some of the largest species on Earth. We tested these hypotheses among marsupials, a group in which mass-specific metabolic rates are roughly 30% lower than those of eutherian mammals of similar size, leading to the expectation that metabolic rate should be a major constraint. Our findings support both hypotheses because levels of sperm competition are associated with increases in sperm size, but low mass-specific metabolic rate constrains sperm size among large species. We also found that the relationship between sperm size and mass-specific metabolic rate is steeper among marsupials and shallower among eutherian mammals. This finding has two implications: marsupials respond to changes in mass-specific metabolic rate by modifying sperm length to a greater extent, suggesting that they are more constrained by metabolic rate. In addition, for any given mass-specific metabolic rate, marsupials produce longer sperm. We suggest that this is the consequence of marsupials diverting resources away from sperm numbers and into sperm size, due to their efficient sperm transport along the female tract and the existence of mechanisms to protect sperm.

  2. Testicular Damage following Testicular Sperm Retrieval

    DEFF Research Database (Denmark)

    Fedder, Jens; Marcussen, Niels; Fedder, Maja D.K.

    2017-01-01

    The aim of this study was to evaluate the possible development of histological abnormalities such as fibrosis and microcalcifications after sperm retrieval in a ram model. Fourteen testicles in nine rams were exposed to open biopsy, multiple TESAs, or TESE, and the remaining four testicles were...... left unoperated on as controls. Three months after sperm retrieval, the testicles were removed, fixed, and cut into 1/2 cm thick slices and systematically put onto a glass plate exposing macroscopic abnormalities. Tissue from abnormal areas was cut into 3 μm sections and stained for histological...... evaluation. Pathological abnormalities were observed in testicles exposed to sperm retrieval (≥11 of 14) compared to 0 of 4 control testicles. Testicular damage was found independently of the kind of intervention used. Therefore, cryopreservation of excess sperm should be considered while retrieving sperm....

  3. BMI in relation to sperm count

    DEFF Research Database (Denmark)

    Sermondade, N; Faure, C; Fezeu, L

    2013-01-01

    BACKGROUND The global obesity epidemic has paralleled a decrease in semen quality. Yet, the association between obesity and sperm parameters remains controversial. The purpose of this report was to update the evidence on the association between BMI and sperm count through a systematic review...... with meta-analysis. METHODS A systematic review of available literature (with no language restriction) was performed to investigate the impact of BMI on sperm count. Relevant studies published until June 2012 were identified from a Pubmed and EMBASE search. We also included unpublished data (n = 717 men...... studies were included in the meta-analysis, resulting in a sample of 13 077 men from the general population and attending fertility clinics. Data were stratified according to the total sperm count as normozoospermia, oligozoospermia and azoospermia. Standardized weighted mean differences in sperm...

  4. Histone H4 Lysine 20 methylation

    DEFF Research Database (Denmark)

    Jørgensen, Stine; Schotta, Gunnar; Sørensen, Claus Storgaard

    2013-01-01

    of histones have emerged as key regulators of genomic integrity. Intense research during the past few years has revealed histone H4 lysine 20 methylation (H4K20me) as critically important for the biological processes that ensure genome integrity, such as DNA damage repair, DNA replication and chromatin...... compaction. The distinct H4K20 methylation states are mediated by SET8/PR-Set7 that catalyses monomethylation of H4K20, whereas SUV4-20H1 and SUV4-20H2 enzymes mediate further H4K20 methylation to H4K20me2 and H4K20me3. Disruption of these H4K20-specific histone methyltransferases leads to genomic...

  5. Recombinant perlucin nucleates the growth of calcium carbonate crystals: molecular cloning and characterization of perlucin from disk abalone, Haliotis discus discus.

    Science.gov (United States)

    Wang, Ning; Lee, Youn-Ho; Lee, Jehee

    2008-02-01

    Perlucin is well known as an important functional protein regulating pearl formation and shell biomineralization. In this study, we cloned the perlucin gene from the abalone Haliotis discus discus cDNA library. The full-length cDNA of the abalone H. discus discus perlucin gene consisted of 1038 bp nucleotides, encoding a putative signal peptide of 22 amino acids and a mature protein of 129 amino acids, which shared 55% identity with the homologous protein in greenlip abalone. The mature protein coding sequence was inserted into pMal-c2X expression vector and it expressed the recombinant protein in E. coli (Rosetta-gammi DE3). The maltose binding protein (MBP) fusion perlucin successfully promoted calcium carbonate precipitation and directed calcite crystal morphological modification. The successful expression of active recombinant perlucin suggested that recombinant perlucin gene transfer has the capability by color modification to improve the pearl's value. In the view of molecular structure, perlucin was a typical C-type lectin, which contained one highly conserved carbohydrate recognition domain. Reverse transcription polymerase chain reaction (RT-PCR) results showed that perlucin gene was expressed not only in the mantle, but also in the gill and digestive tract. Further characterization of perlucin in abalone non-self recognition and disease resistance is promising and anticipated.

  6. Effects of dietary supplementation of probiotic Shewanella colwelliana WA64, Shewanella olleyana WA65 on the innate immunity and disease resistance of abalone, Haliotis discus hannai Ino.

    Science.gov (United States)

    Jiang, Hai-Feng; Liu, Xiao-Lin; Chang, Ya-Qing; Liu, Ming-Tai; Wang, Gao-Xue

    2013-07-01

    The effects of dietary administration of two probiotics, Shewanella colwelliana WA64 and Shewanella olleyana WA65, on the innate immunity of abalone (Haliotis discus hannai Ino), and survival of juvenile abalone challenged with Vibrio harveyi have been studied. Two groups of abalone were fed with three different diets: one control, and two diets supplemented with 10(9) cell g(-1) of probiotic WA64 (WA64 diet) and WA65 (WA65 diet) for up to four weeks. Results showed that abalone fed diets containing S. colwelliana WA64 and S. olleyana WA65 had led to an enhanced cellular and humoral immune response, notably higher haemocytes, respiratory burst activity, serum lysozyme activity and total protein levels were recorded after one week of probiotic administration. On the other hand, mortality after the challenges with V. harveyi in the group fed with control diet ranged from 77 to 80%, while mortality rates observed in the groups fed with diets supplemented with WA64 and WA65 ranged from 27 to 50% and 30-43%, respectively. The results demonstrated potential for S. colwelliana WA64 and S. olleyana WA65 to improve innate immunity and disease resistance in H. discus hannai. Copyright © 2013 Elsevier Ltd. All rights reserved.

  7. Cascading effects from survival to physiological activities, and gene expression of heat shock protein 90 on the abalone Haliotis discus hannai responding to continuous thermal stress.

    Science.gov (United States)

    Park, Kiyun; Lee, Jung Sick; Kang, Ju-Chan; Kim, Jae Won; Kwak, Ihn-Sil

    2015-02-01

    Increasing temperatures can be a significant stressor for aquatic organisms. Abalones, a type of large marine gastropods, are the most commercially important species in aquaculture for Asia. To evaluate the potential ecological risk posed by temperature stress, we measured biological responses such as survival rate, adhesion ability (falling rate), and foot abnormalities in the abalone Haliotis discus hannai. Additionally, biochemical and molecular responses were evaluated in H. discus hannai exposed to various temperature gradients. The survival rate was reduced in abalones exposed to relative high temperatures (more than 26 °C). Increased temperature stress induced a higher falling rate and abnormal foot structure. Furthermore, increased antioxidant enzyme activities were observed in abalones exposed to relative high temperatures (26 and 28 °C). The activities of superoxide dismutase were induced in a time-dependent manner after high temperature stress. Generally, heat shock protein 90 also increased significantly in H. discus hannai exposed to temperature gradients (more than 24 °C) for 12 h. These results provide valuable information regarding stress responses to increased temperatures, in H. discus hannai: adverse biological and molecular outcomes could be utilized as risk assessments and stress monitoring of marine ecosystems under increased water temperatures. Copyright © 2014 Elsevier Ltd. All rights reserved.

  8. [Construction of SSH library from haemocyte of variously colored abalone challenged with bacteria and differential expression analysis of macrophage expressed protein].

    Science.gov (United States)

    Ren, Hong-Lin; Xu, Dan-Dan; Qiao, Kun; Cai, Ling; Huang, Wei-Bin; Zhang, Nai; Wang, Ke-Jian

    2008-08-01

    Abalones are considered to be the most precious delicacy from the sea, and become very important commercial seafood in aquaculture worldwide. Variously colored abalone (Haliotis diversicolor Reeve, 1846) has been widely cultured on the southeast coast for more than twenty years. However, abalone culture frequently suffers from bacterial infection and mass mortality of reared abalones causes serious economic losses. Unfortunately, knowledge of the defense mechanism in this animal is still lacking. In this study, using suppression subtractive hybridization (SSH) technology, a forward SSH library was constructed from haemocytes of H. diversicolor, with the content of 1.37x10(6) pfu and the recombinant rate of 98.18%. After the recombinant plasmids were sequenced, partial cDNA of macrophage expressed protein (MEP) was recognized based on BLAST searches in NCBI, with the size of 1,551 bp, and continuously encoding 517 amino acids. Semi-quantitative PCR and quantitative real-time PCR results showed that MEP cDNA was distinctly up-regulated in haemocytes of the bacterial-challenged group compared to the unchallenged group. The gene information obtained from this library will provide new insights into the immune mechanism of H. diversicolor and facilitate future study of target genes involved in the response to invading microorganisms.

  9. Relationship between Porcine Sperm Motility and Sperm Enzymatic Activity using Paper-based Devices

    Science.gov (United States)

    Matsuura, Koji; Huang, Han-Wei; Chen, Ming-Cheng; Chen, Yu; Cheng, Chao-Min

    2017-04-01

    Mammalian sperm motility has traditionally been analyzed to determine fertility using computer-assisted semen analysis (CASA) systems. To develop low-cost and robust male fertility diagnostics, we created a paper-based MTT assay and used it to estimate motile sperm concentration. When porcine sperm motility was inhibited using sperm enzyme inhibitors for sperm enzymes related to mitochondrial activity and glycolysis, we simultaneously recorded sperm motility and enzymatic reactivity using a portable motility analysis system (iSperm) and a paper-based MTT assay, respectively. When using our paper-based MTT-assay, we calculated the area mean value signal intensity (AMV) to evaluate enzymatic reactivity. Both sperm motility and AMV decreased following treatment with iodoacetamide (IODO) and 3-bromopyruvic acid (3BP), both of which are inhibitors of glycolytic enzymes including glyceraldehyde-3-phosphate dehydrogenase (GAPDH). We found a correlation between recorded motility using iSperm and AMV from our paper-based assay (P coupled with catalysis of GAPDH and was promoted by electron transfer from NADH. Based on this inhibitor study, sperm motility can be estimated using our paper-based MTT-assay.

  10. The influence of benign prostatic hyperplasia on sperm morphological features and sperm DNA integrity in dogs.

    Science.gov (United States)

    Flores, R B; Angrimani, Dsr; Rui, B R; Brito, M M; Abreu, R A; Vannucchi, C I

    2017-04-01

    Benign prostatic hyperplasia (BPH) has a high incidence in older intact dogs. Due to the increased prostatic oxidative stress and hormonal imbalance of BPH, sperm damage can arise, such as sperm morphological alterations and DNA fragmentation. This study aimed to compare the reproductive potential of healthy dogs and those affected by benign prostatic hyperplasia. Ten dogs were assigned to two experimental groups: dogs without BPH (control; n = 5) and dogs diagnosed with BPH (n = 5), based on clinical signs and ultrasonographic findings. Three semen collections were performed from each dog within one month and analysed using computer-assisted sperm analysis (CASA) and functional tests. Control group showed higher percentage of sperm DNA integrity (95 ± 1.8%) compared to the BPH group (79.2 ± 6.4%). On the other hand, the percentage of minor sperm defects, amplitude of lateral sperm head displacement of the spermatozoa and medium sperm mitochondrial activity were higher in the BPH group. In conclusion, BPH decreases sperm DNA integrity, increases mitochondrial activity, as well as modifies sperm movement pattern. Therefore, a careful sperm analysis of aged dogs with BPH is required before a reproductive programme can be established for such patients. © 2016 Blackwell Verlag GmbH.

  11. Lysine Acetylation and Deacetylation in Brain Development and Neuropathies.

    Science.gov (United States)

    Tapias, Alicia; Wang, Zhao-Qi

    2017-02-01

    Embryonic development is critical for the final functionality and maintenance of the adult brain. Brain development is tightly regulated by intracellular and extracellular signaling. Lysine acetylation and deacetylation are posttranslational modifications that are able to link extracellular signals to intracellular responses. A wealth of evidence indicates that lysine acetylation and deacetylation are critical for brain development and functionality. Indeed, mutations of the enzymes and cofactors responsible for these processes are often associated with neurodevelopmental and psychiatric disorders. Lysine acetylation and deacetylation are involved in all levels of brain development, starting from neuroprogenitor survival and proliferation, cell fate decisions, neuronal maturation, migration, and synaptogenesis, as well as differentiation and maturation of astrocytes and oligodendrocytes, to the establishment of neuronal circuits. Hence, fluctuations in the balance between lysine acetylation and deacetylation contribute to the final shape and performance of the brain. In this review, we summarize the current basic knowledge on the specific roles of lysine acetyltransferase (KAT) and lysine deacetylase (KDAC) complexes in brain development and the different neurodevelopmental disorders that are associated with dysfunctional lysine (de)acetylation machineries. Copyright © 2017 The Authors. Production and hosting by Elsevier Ltd.. All rights reserved.

  12. Myeloperoxidase-mediated protein lysine oxidation generates 2-aminoadipic acid and lysine nitrile in vivo.

    Science.gov (United States)

    Lin, Hongqiao; Levison, Bruce S; Buffa, Jennifer A; Huang, Ying; Fu, Xiaoming; Wang, Zeneng; Gogonea, Valentin; DiDonato, Joseph A; Hazen, Stanley L

    2017-03-01

    Recent studies reveal 2-aminoadipic acid (2-AAA) is both elevated in subjects at risk for diabetes and mechanistically linked to glucose homeostasis. Prior studies also suggest enrichment of protein-bound 2-AAA as an oxidative post-translational modification of lysyl residues in tissues associated with degenerative diseases of aging. While in vitro studies suggest redox active transition metals or myeloperoxidase (MPO) generated hypochlorous acid (HOCl) may produce protein-bound 2-AAA, the mechanism(s) responsible for generation of 2-AAA during inflammatory diseases are unknown. In initial studies we observed that traditional acid- or base-catalyzed protein hydrolysis methods previously employed to measure tissue 2-AAA can artificially generate protein-bound 2-AAA from an alternative potential lysine oxidative product, lysine nitrile (LysCN). Using a validated protease-based digestion method coupled with stable isotope dilution LC/MS/MS, we now report protein bound 2-AAA and LysCN are both formed by hypochlorous acid (HOCl) and the MPO/H 2 O 2 /Cl - system of leukocytes. At low molar ratio of oxidant to target protein N ε -lysine moiety, 2-AAA is formed via an initial N ε -monochloramine intermediate, which ultimately produces the more stable 2-AAA end-product via sequential generation of transient imine and semialdehyde intermediates. At higher oxidant to target protein N ε -lysine amine ratios, protein-bound LysCN is formed via initial generation of a lysine N ε -dichloramine intermediate. In studies employing MPO knockout mice and an acute inflammation model, we show that both free and protein-bound 2-AAA, and in lower yield, protein-bound LysCN, are formed by MPO in vivo during inflammation. Finally, both 2-AAA and to lesser extent LysCN are shown to be enriched in human aortic atherosclerotic plaque, a tissue known to harbor multiple MPO-catalyzed protein oxidation products. Collectively, these results show that MPO-mediated oxidation of protein lysyl

  13. Office-based sperm concentration: A simplified method for ...

    African Journals Online (AJOL)

    Methods: Semen samples from 51 sperm donors were used. Following swim-up separation, the sperm concentration of the retrieved motile fraction was counted, as well as progressive motile sperm using a standardised wet preparation. The number of sperm in a 10 μL droplet covered with a 22 × 22 mm coverslip was ...

  14. Tenacibaculum haliotis sp. nov., isolated from the gut of an abalone Haliotis discus hannai.

    Science.gov (United States)

    Kim, Young-Ok; Park, In-Suk; Park, Sooyeon; Nam, Bo-Hye; Park, Ji-Min; Kim, Dong-Gyun; Yoon, Jung-Hoon

    2017-09-01

    A Gram-stain-negative, non-flagellated, gliding, non-spore-forming bacterial strain, designated RA3-2T, was isolated from the gut of an abalone (Haliotis discus hannai) collected from the sea around Jeju island, South Korea, and subjected to a polyphasic taxonomic study. RA3-2T grew optimally at 20 °C and in the presence of 2.0-3.0 % (w/v) NaCl. Phylogenetic trees based on 16S rRNA gene sequences indicated that RA3-2T fell within the clade comprising the type strains of species of the genus Tenacibaculum, clustering with the type strains of Tenacibaculum soleae, Tenacibaculum ovolyticum and Tenacibaculum dicentrarchi; showing 16S rRNA gene sequence similarity values of 96.2-96.8 %. The novel strain exhibited 16S rRNA gene sequence similarity values of 93.5-96.9 % to the type strains of the other species of the genus Tenacibaculum. RA3-2T contained MK-6 as the predominant menaquinone and iso-C15 : 0 and iso-C15 : 0 3-OH as the major fatty acids. The major polar lipids of RA3-2T were phosphatidylethanolamine, two unidentified lipids, one unidentified aminophospholipid and one unidentified glycolipid. The DNA G+C content of RA3-2T was 31.7 mol%. The differential phenotypic properties, together with the phylogenetic data, revealed that RA3-2T is separated from other species of the genus Tenacibaculum with validly published names. On the basis of the data presented, RA3-2T is considered to represent a novel species of the genus Tenacibaculum, for which the name Tenacibaculum haliotis sp. nov. is proposed. The type strain is RA3-2T (=KCTC 52419T=NBRC 112382T).

  15. Exploring Spatiotemporal Trends in Commercial Fishing Effort of an Abalone Fishing Zone: A GIS-Based Hotspot Model.

    Directory of Open Access Journals (Sweden)

    M Ali Jalali

    Full Text Available Assessing patterns of fisheries activity at a scale related to resource exploitation has received particular attention in recent times. However, acquiring data about the distribution and spatiotemporal allocation of catch and fishing effort in small scale benthic fisheries remains challenging. Here, we used GIS-based spatio-statistical models to investigate the footprint of commercial diving events on blacklip abalone (Haliotis rubra stocks along the south-west coast of Victoria, Australia from 2008 to 2011. Using abalone catch data matched with GPS location we found catch per unit of fishing effort (CPUE was not uniformly spatially and temporally distributed across the study area. Spatial autocorrelation and hotspot analysis revealed significant spatiotemporal clusters of CPUE (with distance thresholds of 100's of meters among years, indicating the presence of CPUE hotspots focused on specific reefs. Cumulative hotspot maps indicated that certain reef complexes were consistently targeted across years but with varying intensity, however often a relatively small proportion of the full reef extent was targeted. Integrating CPUE with remotely-sensed light detection and ranging (LiDAR derived bathymetry data using generalized additive mixed model corroborated that fishing pressure primarily coincided with shallow, rugose and complex components of reef structures. This study demonstrates that a geospatial approach is efficient in detecting patterns and trends in commercial fishing effort and its association with seafloor characteristics.

  16. Habitat Preferences of Juvenile Abalone (Haliotis mariae Wood, 1828 Along the Dhofar Coast of Oman and Implications for Conservation

    Directory of Open Access Journals (Sweden)

    Schalk Willem Petrus de Waal

    2012-01-01

    Full Text Available Surveys were conducted along the eastern Dhofar coast of Oman to investigate densities and habitat preferences of juvenile Haliotis mariae ( 50 cm Ø, respectively. B values were highest for urchins (6 times that for small boulders, and for boulders <50 cm Ø. B values for boulder habitats decreased as boulder size increased. Urchin utilisation by juvenile abalone as shelter ranged between geographic areas from a minimum of 15.5% to a maximum of 47.6%. The proportion of total habitat that is preferred by more than 97% of juvenile abalone found, including urchins and boulders < 50 cm Ø, comprises 29% of surveyed substratum. While the role urchins play on wild juvenile H. mariae has not proved vital, it is definitely significant. Although juvenile densities are low and are not currently limited by the availability of suitable habitat, it is crucial to identify and conserve those microhabitats that support recruitment of H. mariae. The abundance of these areas should be among the criteria used in selecting protected conservation areas.

  17. Tumor suppressor QM-like gene from disk abalone (Haliotis discus discus): molecular characterization and transcriptional analysis upon immune challenge.

    Science.gov (United States)

    Oh, Chulhong; De Zoysa, Mahanama; Nikapitiya, Chamilani; Whang, Ilson; Kim, Yu Cheol; Kang, Do-Hyung; Heo, Soo-jin; Choi, Young-Ung; Choi, Cheol Young; Lee, Jae-Seong; Lee, Jehee

    2010-09-01

    We describe molecular characterization and transcriptional analysis of the gene encoding tumor suppressor QM-like protein, AbQM, in the disk abalone Haliotis discus discus. The full-length cDNA (765-bp) of AbQM was found to consist of a 654-bp ORF coding for a 218 amino acid protein of a 25 kDa molecular mass with a 10.2 isoelectric point. Analysis of AbQM sequence revealed the presence of characteristic motifs, including the ribosomal protein L10 signature, SH3-binding motif and two antibiotic binding sites. Phylogenetic analysis confirmed that AbQM is closely related to other mollusk QM proteins, and altogether they form a mollusk QM protein sub-family which displays evolutionary conservation from yeast to human. Tissue-specific expression and transcriptional regulation of AbQM was analyzed by quantitative real-time PCR in response to bacterial (Vibrio alginolyticus and Vibrio parahemolyticus, Listeria monocytogenes) and viral (viral hemorrhagic septicemia virus, VHSV) challenge. AbQM transcripts were found to be expressed ubiquitously in all examined tissues in a constitutive manner, as similar expression levels were detected in hemocytes, mantle, digestive tract and muscle. Upon bacterial and VHSV challenge, AbQM showed significant up-regulation in gills, but not in hemocytes. Taken together, these findings suggest that AbQM in abalone-like mollusks can respond to and facilitate a defensive effect against pathogenic infection. Copyright 2010 Elsevier Ltd. All rights reserved.

  18. Seasonal and algal diet-driven patterns of the digestive microbiota of the European abalone Haliotis tuberculata, a generalist marine herbivore.

    Science.gov (United States)

    Gobet, Angélique; Mest, Laëtitia; Perennou, Morgan; Dittami, Simon M; Caralp, Claire; Coulombet, Céline; Huchette, Sylvain; Roussel, Sabine; Michel, Gurvan; Leblanc, Catherine

    2018-03-27

    Holobionts have a digestive microbiota with catabolic abilities allowing the degradation of complex dietary compounds for the host. In terrestrial herbivores, the digestive microbiota is known to degrade complex polysaccharides from land plants while in marine herbivores, the digestive microbiota is poorly characterized. Most of the latter are generalists and consume red, green, and brown macroalgae, three distinct lineages characterized by a specific composition in complex polysaccharides, which represent half of their biomass. Subsequently, each macroalga features a specific epiphytic microbiota, and the digestive microbiota of marine herbivores is expected to vary with a monospecific algal diet. We investigated the effect of four monospecific diets (Palmaria palmata, Ulva lactuca, Saccharina latissima, Laminaria digitata) on the composition and specificity of the digestive microbiota of a generalist marine herbivore, the abalone, farmed in a temperate coastal area over a year. The microbiota from the abalone digestive gland was sampled every 2 months and explored using metabarcoding. Diversity and multivariate analyses showed that patterns of the microbiota were significantly linked to seasonal variations of contextual parameters but not directly to a specific algal diet. Three core genera: Psychrilyobacter, Mycoplasma, and Vibrio constantly dominated the microbiota in the abalone digestive gland. Additionally, a less abundant and diet-specific core microbiota featured genera representing aerobic primary degraders of algal polysaccharides. This study highlights the establishment of a persistent core microbiota in the digestive gland of the abalone since its juvenile state and the presence of a less abundant and diet-specific core community. While composed of different microbial taxa compared to terrestrial herbivores, the digestive gland constitutes a particular niche in the abalone holobiont, where bacteria (i) may cooperate to degrade algal polysaccharides to

  19. Advanced sperm selection techniques for assisted reproduction.

    Science.gov (United States)

    McDowell, Simon; Kroon, Ben; Ford, Emily; Hook, Ysanne; Glujovsky, Demián; Yazdani, Anusch

    2014-10-28

    Assisted reproductive technologies (ART) such as in vitro fertilisation (IVF) and intracytoplasmic sperm injection (ICSI) bring together gametes outside of the body to enhance the probability of fertilisation and pregnancy. Advanced sperm selection techniques are increasingly being employed in ART, most commonly in cycles utilising ICSI. Advanced sperm selection techniques are thought to improve the chance that structurally intact and mature sperm with high DNA integrity are selected for fertilisation. Advanced sperm selection strategies include selection according to surface charge; sperm apoptosis; sperm birefringence; ability to bind to hyaluronic acid; and sperm morphology under ultra-high magnification. These techniques theoretically improve ART outcomes. To evaluate the impact of advanced sperm selection techniques on ART outcomes. Systematic search of electronic databases (Cochrane Menstrual Disorders and Subfertility Group Specialised Register, the Cochrane Central Register of Controlled Trials (CENTRAL), MEDLINE, EMBASE, PsycINFO, Cumulative Index to Nursing and Allied Health Literature (CINAHL), Latin American and Caribbean Health Science Information Database (LILACS)), trials registers (ClinicalTrials.gov, Current Controlled Trials, World Health Organization International Clinical Trials Registry Platform), conference abstracts (Web of Knowledge) and grey literature (OpenGrey) for relevant randomised controlled trials. We handsearched the reference lists of included studies and similar reviews. The search was conducted in May 2014. We included randomised controlled trials (RCTs) comparing an advanced sperm selection technique versus standard IVF or ICSI or versus another advanced sperm selection technique. We excluded studies of sperm selection using ultra-high magnification (intracytoplasmic morphologically selected sperm injection, or IMSI), as they are the subject of a separate Cochrane review. Quasi-randomised and pseudo-randomised trials were

  20. Redox regulation of mammalian sperm capacitation

    Directory of Open Access Journals (Sweden)

    Cristian O′Flaherty

    2015-01-01

    Full Text Available Capacitation is a series of morphological and metabolic changes necessary for the spermatozoon to achieve fertilizing ability. One of the earlier happenings during mammalian sperm capacitation is the production of reactive oxygen species (ROS that will trigger and regulate a series of events including protein phosphorylation, in a time-dependent fashion. The identity of the sperm oxidase responsible for the production of ROS involved in capacitation is still elusive, and several candidates are discussed in this review. Interestingly, ROS-induced ROS formation has been described during human sperm capacitation. Redox signaling during capacitation is associated with changes in thiol groups of proteins located on the plasma membrane and subcellular compartments of the spermatozoon. Both, oxidation of thiols forming disulfide bridges and the increase on thiol content are necessary to regulate different sperm proteins associated with capacitation. Reducing equivalents such as NADH and NADPH are necessary to support capacitation in many species including humans. Lactate dehydrogenase, glucose-6-phospohate dehydrogenase, and isocitrate dehydrogenase are responsible in supplying NAD (P H for sperm capacitation. Peroxiredoxins (PRDXs are newly described enzymes with antioxidant properties that can protect mammalian spermatozoa; however, they are also candidates for assuring the regulation of redox signaling required for sperm capacitation. The dysregulation of PRDXs and of enzymes needed for their reactivation such as thioredoxin/thioredoxin reductase system and glutathione-S-transferases impairs sperm motility, capacitation, and promotes DNA damage in spermatozoa leading to male infertility.

  1. Sperm Preservation using Freeze-Drying Method

    Directory of Open Access Journals (Sweden)

    TAKDIR SAILI

    2005-03-01

    Full Text Available Since the discovery of cryopreservation method for bull semen, cryopreservation become an alternative method for maintaining gamet resources of certain animal which is threatened or near extinction. This technology was then applied to the preservation of embryo, oocyte, ovary and testis. The application of intracytoplasmic sperm injection (ICSI for which sperm motility is unnecessary had supported the effort to create simplified method such as freeze-drying for sperm preservation. Due to the benefit of ICSI over the conventional in vitro fertilization (IVF the spermatozoon could be mechanically driven to pass through the zona pellucida and entering the cytoplasm of oocytes prior to fertilization. The freeze-drying method is an alternative method in sperm preservation which ignored the motility of sperm. The sperm resulted from this technique is in drying state, therefore, it might be stored in room temperature or in refrigerator. Many reports have claimed that freeze-dried sperm which is not motile but has an intact DNA was able to fertilize oocytes, even produced offspring in mouse.

  2. Redox regulation of mammalian sperm capacitation

    Science.gov (United States)

    O’Flaherty, Cristian

    2015-01-01

    Capacitation is a series of morphological and metabolic changes necessary for the spermatozoon to achieve fertilizing ability. One of the earlier happenings during mammalian sperm capacitation is the production of reactive oxygen species (ROS) that will trigger and regulate a series of events including protein phosphorylation, in a time-dependent fashion. The identity of the sperm oxidase responsible for the production of ROS involved in capacitation is still elusive, and several candidates are discussed in this review. Interestingly, ROS-induced ROS formation has been described during human sperm capacitation. Redox signaling during capacitation is associated with changes in thiol groups of proteins located on the plasma membrane and subcellular compartments of the spermatozoon. Both, oxidation of thiols forming disulfide bridges and the increase on thiol content are necessary to regulate different sperm proteins associated with capacitation. Reducing equivalents such as NADH and NADPH are necessary to support capacitation in many species including humans. Lactate dehydrogenase, glucose-6-phospohate dehydrogenase, and isocitrate dehydrogenase are responsible in supplying NAD (P) H for sperm capacitation. Peroxiredoxins (PRDXs) are newly described enzymes with antioxidant properties that can protect mammalian spermatozoa; however, they are also candidates for assuring the regulation of redox signaling required for sperm capacitation. The dysregulation of PRDXs and of enzymes needed for their reactivation such as thioredoxin/thioredoxin reductase system and glutathione-S-transferases impairs sperm motility, capacitation, and promotes DNA damage in spermatozoa leading to male infertility. PMID:25926608

  3. Predicting post-translational lysine acetylation using support vector machines

    DEFF Research Database (Denmark)

    Gnad, Florian; Ren, Shubin; Choudhary, Chunaram

    2010-01-01

    Lysine acetylation is a post-translational protein modification and a primary regulatory mechanism that controls many cell signaling processes. Lysine acetylation sites are recognized by acetyltransferases and deacetylases through sequence patterns (motifs). Recently, we used high-resolution mass...... spectrometry to identify 3600 lysine acetylation sites on 1750 human proteins covering most of the previously annotated sites and providing the most comprehensive acetylome so far. This dataset should provide an excellent source to train support vector machines (SVMs) allowing the high accuracy in silico...

  4. Effects of dietary chromium tripicolinate and lysine on growth ...

    African Journals Online (AJOL)

    A la fin de l'essai, les taux des acides gras non estérifiés ont été élevés par la lysine (lys quadratique, p < 0,08), de même que ceux des protéines totales (lys quadratique, p < 0,02). Les valeurs de l'azote de l'urée étaient également élevées par la lysine (lys linéaire, p < 0,0002). L'effet de l'interaction de CrPic et de la lysine

  5. HDAC inhibitors induce global changes in histone lysine and arginine methylation and alter expression of lysine demethylases.

    Science.gov (United States)

    Lillico, Ryan; Sobral, Marina Gomez; Stesco, Nicholas; Lakowski, Ted M

    2016-02-05

    Histone deacetylase (HDAC) inhibitors are cancer treatments that inhibit the removal of the epigenetic modification acetyllysine on histones, resulting in altered gene expression. Such changes in expression may influence other histone epigenetic modifications. We describe a validated liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to quantify lysine acetylation and methylation and arginine methylation on histones extracted from cultured cells treated with HDAC inhibitors. The HDAC inhibitors vorinostat, mocetinostat and entinostat induced 400-600% hyperacetylation in HEK 293 and K562 cells. All HDAC inhibitors decreased histone methylarginines in HEK 293 cells but entinostat produced dose dependent reductions in asymmetric dimethylarginine, not observed in K562 cells. Vorinostat produced increases in histone lysine methylation and decreased expression of some lysine demethylases (KDM), measured by quantitative PCR. Entinostat had variable effects on lysine methylation and decreased expression of some KDM while increasing expression of others. Mocetinostat produced dose dependent increases in histone lysine methylation by LC-MS/MS. This was corroborated with a multiplex colorimetric assay showing increases in histone H3 lysine 4, 9, 27, 36 and 79 methylation. Increases in lysine methylation were correlated with dose dependent decreases in the expression of seven KDM. Mocetinostat functions as an HDAC inhibitor and a de facto KDM inhibitor. Copyright © 2015 Elsevier B.V. All rights reserved.

  6. The stochastic dance of circling sperm cells: sperm chemotaxis in the plane

    Energy Technology Data Exchange (ETDEWEB)

    Friedrich, B M; Juelicher, F [Max Planck Institute for the Physics of Complex Systems, Noethnitzer Strasse 38, 01187 Dresden (Germany)], E-mail: ben@pks.mpg.de, E-mail: julicher@pks.mpg.de

    2008-12-15

    Biological systems such as single cells must function in the presence of fluctuations. It has been shown in a two-dimensional experimental setup that sea urchin sperm cells move toward a source of chemoattractant along planar trochoidal swimming paths, i.e. drifting circles. In these experiments, a pronounced variability of the swimming paths is observed. We present a theoretical description of sperm chemotaxis in two dimensions which takes fluctuations into account. We derive a coarse-grained theory of stochastic sperm swimming paths in a concentration field of chemoattractant. Fluctuations enter as multiplicative noise in the equations for the sperm swimming path. We discuss the stochastic properties of sperm swimming and predict a concentration-dependence of the effective diffusion constant of sperm swimming which could be tested in experiments.

  7. The stochastic dance of circling sperm cells: sperm chemotaxis in the plane

    Science.gov (United States)

    Friedrich, B. M.; Jülicher, F.

    2008-12-01

    Biological systems such as single cells must function in the presence of fluctuations. It has been shown in a two-dimensional experimental setup that sea urchin sperm cells move toward a source of chemoattractant along planar trochoidal swimming paths, i.e. drifting circles. In these experiments, a pronounced variability of the swimming paths is observed. We present a theoretical description of sperm chemotaxis in two dimensions which takes fluctuations into account. We derive a coarse-grained theory of stochastic sperm swimming paths in a concentration field of chemoattractant. Fluctuations enter as multiplicative noise in the equations for the sperm swimming path. We discuss the stochastic properties of sperm swimming and predict a concentration-dependence of the effective diffusion constant of sperm swimming which could be tested in experiments.

  8. How accurate is sperm morphology as an indicator of sperm function?

    Science.gov (United States)

    Franken, D R

    2015-08-01

    Sperm morphology has been consistently correlated with fertilisation success or failure. The clinical relevance of the percentage normal spermatozoa has been a widely discussed topic amongst infertility specialists and scientists. This study aimed to evaluate the role of sperm morphology as an indicator of additional sperm functions among 114 andrology referrals. The sperm functions that were investigated included chromatin packaging quality (CMA3 test (n = 109), zona-induced acrosome reaction (ZIAR test; n = 36), hemizona assay (HZI; n = 36) and progressive motility (n = 47). Chromatin packaging quality had a negative and significant (P = 0.0001, r = -0.74) correlation with the percentage normal spermatozoa, while progressive motility had a significant and positive correlation (P = 0.0001, 0.59). Accurate sperm morphology scoring as described by the WHO 2010 manual can therefore be used as an indicator of specific sperm functions. © 2014 Blackwell Verlag GmbH.

  9. Tenacity of exogenous human papillomavirus DNA in sperm washing.

    Science.gov (United States)

    Brossfield, J E; Chan, P J; Patton, W C; King, A

    1999-07-01

    Sperm cells have been shown to take up exogenous DNA readily. The hypothesis was that sperm washing would remove exogenous viral DNA infecting sperm cells. The objective was to compare three types of sperm washing procedures for their capacity to remove exogenous human papillomavirus (HPV) DNA from infected sperm. Prewashed sperm were equally divided and sperm in one portion were exposed to L1 HPV DNA fragments for 30 min at 37 degrees C. Untreated washed sperm served as the control. After transfection, the sperm were washed by either centrifuge, two-layer Isolate colloid wash, or test-yolk buffer procedures. Sperm parameters were measured on a Hamilton Thorn HTM-C analyzer. Sperm DNA were extracted and polymerase chain reaction (PCR) was carried out targeting the L1 consensus gene of HPV and the designated sentinel gene, 17q21 spanning the D17S855 gene. Amplified products were analyzed in 2% agarose gel electrophoresis. PCR analyses detected the consensus L1 HPV gene in sperm after they were processed through either of the three procedures. Controls were negative for the L1 gene. Extracted DNA were verified by PCR amplification of 17q21 spanning the D17S855 gene. Transfected sperm had higher percentages of total motility and progression compared with the control. Centrifuged, washed, transfected sperm exhibited a greater curvilinear velocity and hyperactivation. The data showed that washing would not remove exogenous HPV DNA from sperm cells. The viral DNA was tenaciously bound to the sperm, suggesting an internalization into the sperm. The viral DNA also increased the motility of the sperm by affecting the velocity and progression of the sperm, which suggested either an increase in metabolism, an enhancement of the calcium-regulated motility mechanism, or an artifact of PCR reagents. More studies are needed to elucidate the mechanism of DNA stimulated sperm motility.

  10. Direct action of endocrine disrupting chemicals on human sperm

    DEFF Research Database (Denmark)

    Schiffer, Christian; Müller, Astrid; Egeberg, Dorte L

    2014-01-01

    sperm. We show that structurally diverse EDCs activate the sperm-specific CatSper channel and, thereby, evoke an intracellular Ca(2+) increase, a motility response, and acrosomal exocytosis. Moreover, EDCs desensitize sperm for physiological CatSper ligands and cooperate in low-dose mixtures to elevate...... Ca(2+) levels in sperm. We conclude that EDCs interfere with various sperm functions and, thereby, might impair human fertilization....

  11. Sperm immobilization by dental focus microorganisms.

    Science.gov (United States)

    Linossier, A; Thumann, A; Bustos-Obregon, E

    1982-01-01

    Focal infections and their ability to produce alterations in different tissues have been in dispute for long time. The purpose of this work was to observe "in vitro" the effect of an Escherichia coli filtrate obtained from open pulpar necrosis on human sperm motility. It was observed that the E. coli filtrate produced a loss in sperm motility. The immobilizating factor was studied and characterized as a heat-stable, resistant to lyophilization and non-dializable substance, which could via blood stream reach the male reproductive system and affect sperm motility.

  12. Ovarian fluid mediates the temporal decline in sperm viability in a fish with sperm storage.

    Directory of Open Access Journals (Sweden)

    Clelia Gasparini

    Full Text Available A loss of sperm viability and functionality during sperm transfer and storage within the female reproductive tract can have important fitness implications by disrupting fertilization and impairing offspring development and survival. Consequently, mechanisms that mitigate the temporal decline in sperm function are likely to be important targets of selection. In many species, ovarian fluid is known to regulate and maintain sperm quality. In this paper, we use the guppy Poecilia reticulata, a highly polyandrous freshwater fish exhibiting internal fertilization and sperm storage, to determine whether ovarian fluid (OF influences the decline in sperm viability (the proportion of live sperm in the ejaculate over time and whether any observed effects depend on male sexual ornamentation. To address these questions we used a paired experimental design in which ejaculates from individual males were tested in vitro both in presence and absence of OF. Our results revealed that the temporal decline in sperm viability was significantly reduced in the presence of OF compared to a saline control. This finding raises the intriguing possibility that OF may play a role in mediating the decline in sperm quality due to the deleterious effects of sperm ageing, although other possible explanations for this observation are discussed. Interestingly, we also show that the age-related decline in sperm viability was contingent on male sexual ornamentation; males with relatively high levels of iridescence (indicating higher sexual attractiveness exhibited a more pronounced decline in sperm viability over time than their less ornamented counterparts. This latter finding offers possible insights into the functional basis for the previously observed trade-off between these key components of pre- and postcopulatory sexual selection.

  13. Effect of vitamins and bivalent metals on lysine yield in Bacillus ...

    African Journals Online (AJOL)

    The effects of vitamins and bivalent metals on lysine accumulation in Bacillus strains were investigated. Biotin enhanced lysine production in all the Bacillus strains, while folic acid and riboflavin stimulated lysine yields in Bacillus megaterium SP 86 only. All bivalent metals stimulated lysine accumulation in B. megaterium ...

  14. Enzymatic production of 5-aminovalerate from l-lysine using l-lysine monooxygenase and 5-aminovaleramide amidohydrolase

    OpenAIRE

    Liu, Pan; Zhang, Haiwei; Lv, Min; Hu, Mandong; Li, Zhong; Gao, Chao; Xu, Ping; Ma, Cuiqing

    2014-01-01

    5-Aminovalerate is a potential C5 platform chemical for synthesis of valerolactam, 5-hydroxyvalerate, glutarate, and 1,5-pentanediol. It is a metabolite of l-lysine catabolism through the aminovalerate pathway in Pseudomonas putida. l-Lysine monooxygenase (DavB) and 5-aminovaleramide amidohydrolase (DavA) play key roles in the biotransformation of l-lysine into 5-aminovalerate. Here, DavB and DavA of P. putida KT2440 were expressed, purified, and coupled for the production of 5-aminovalerate ...

  15. Use of testicular sperm for intracytoplasmic sperm injection in men with high sperm DNA fragmentation: a SWOT analysis

    Science.gov (United States)

    Esteves, Sandro C; Roque, Matheus; Garrido, Nicolás

    2018-01-01

    Spermatozoa retrieved from the testis of men with high levels of sperm DNA fragmentation (SDF) in the neat semen tend to have better DNA quality. Given the negative impact of SDF on the outcomes of Assisted Reproductive Technology (ART), an increased interest has emerged about the use of testicular sperm for intracytoplasmic sperm injection (Testi-ICSI). In this article, we used a SWOT (strengths, weaknesses, opportunities, and threats) analysis to summarize the advantages and drawbacks of this intervention. The rationale of Testi-ICSI is bypass posttesticular DNA fragmentation caused by oxidative stress during sperm transit through the epididymis. Hence, oocyte fertilization by genomically intact testicular spermatozoa may be optimized, thus increasing the chances of creating a normal embryonic genome and the likelihood of achieving a live birth, as recently demonstrated in men with high SDF. However, there is still limited evidence as regards the clinical efficacy of Testi-ICSI, thus creating opportunities for further confirmatory clinical research as well as investigation of Testi-ICSI in clinical scenarios other than high SDF. Furthermore, Testi-ICSI can be compared to other laboratory preparation methods for deselecting sperm with damaged DNA. At present, the available literature supports the use of testicular sperm when performing ICSI in infertile couples whose male partners have posttesticular SDF. Due to inherent risks of sperm retrieval, Testi-ICSI should be offered when less invasive treatments for alleviating DNA damage have failed. A call for continuous monitoring is nonetheless required concerning the health of generated offspring and the potential complications of sperm retrieval. PMID:28440264

  16. Use of testicular sperm for intracytoplasmic sperm injection in men with high sperm DNA fragmentation: a SWOT analysis.

    Science.gov (United States)

    Esteves, Sandro C; Roque, Matheus; Garrido, Nicolás

    2018-01-01

    Spermatozoa retrieved from the testis of men with high levels of sperm DNA fragmentation (SDF) in the neat semen tend to have better DNA quality. Given the negative impact of SDF on the outcomes of Assisted Reproductive Technology (ART), an increased interest has emerged about the use of testicular sperm for intracytoplasmic sperm injection (Testi-ICSI). In this article, we used a SWOT (strengths, weaknesses, opportunities, and threats) analysis to summarize the advantages and drawbacks of this intervention. The rationale of Testi-ICSI is bypass posttesticular DNA fragmentation caused by oxidative stress during sperm transit through the epididymis. Hence, oocyte fertilization by genomically intact testicular spermatozoa may be optimized, thus increasing the chances of creating a normal embryonic genome and the likelihood of achieving a live birth, as recently demonstrated in men with high SDF. However, there is still limited evidence as regards the clinical efficacy of Testi-ICSI, thus creating opportunities for further confirmatory clinical research as well as investigation of Testi-ICSI in clinical scenarios other than high SDF. Furthermore, Testi-ICSI can be compared to other laboratory preparation methods for deselecting sperm with damaged DNA. At present, the available literature supports the use of testicular sperm when performing ICSI in infertile couples whose male partners have posttesticular SDF. Due to inherent risks of sperm retrieval, Testi-ICSI should be offered when less invasive treatments for alleviating DNA damage have failed. A call for continuous monitoring is nonetheless required concerning the health of generated offspring and the potential complications of sperm retrieval.

  17. Motile sperm organelle morphology examination (MSOME) and sperm head vacuoles: state of the art in 2013.

    Science.gov (United States)

    Perdrix, Anne; Rives, Nathalie

    2013-01-01

    BACKGROUND Approximately 10 years after the first publication introducing the motile sperm organelle morphology examination (MSOME), many questions remained about sperm vacuoles: frequency, size, localization, mode of occurrence, biological significance and impact on male fertility potential. Many studies have tried to characterize sperm vacuoles, to determine the sperm abnormalities possibly associated with vacuoles, to test the diagnostic value of MSOME for male infertility or to question the benefits of intracytoplasmic morphologically selected sperm injection (IMSI). METHODS We searched PubMed for articles in the English language published in 2001-2012 regarding human sperm head vacuoles, MSOME and IMSI. RESULTS A bibliographic analysis revealed consensus for the following findings: (i) sperm vacuoles appeared frequently, often multiple and preferentially anterior; (ii) sperm vacuoles and sperm chromatin immaturity have been associated, particularly in the case of large vacuoles; (iii) teratozoospermia was a preferred indication of MSOME and IMSI. CONCLUSION The high-magnification system appears to be a powerful method to improve our understanding of human spermatozoa. However, its clinical use remains unclear in the fields of male infertility diagnosis and assisted reproduction techniques (ARTs).

  18. Importance of sperm morphology during sperm transport and fertilization in mammals

    Directory of Open Access Journals (Sweden)

    Francisco A García-Vázquez

    2016-01-01

    Full Text Available After natural or artificial insemination, the spermatozoon starts a journey from the site of deposition to the place of fertilization. However, only a small subset of the spermatozoa deposited achieves their goal: to reach and fertilize the egg. Factors involved in controlling sperm transport and fertilization include the female reproductive tract environment, cell-cell interactions, gene expression, and phenotypic sperm traits. Some of the significant determinants of fertilization are known (i.e., motility or DNA status, but many sperm traits are still indecipherable. One example is the influence of sperm dimensions and shape upon transport within the female genital tract towards the oocyte. Biophysical associations between sperm size and motility may influence the progression of spermatozoa through the female reproductive tract, but uncertainties remain concerning how sperm morphology influences the fertilization process, and whether only the sperm dimensions per se are involved. Moreover, such explanations do not allow the possibility that the female tract is capable of distinguishing fertile spermatozoa on the basis of their morphology, as seems to be the case with biochemical, molecular, and genetic properties. This review focuses on the influence of sperm size and shape in evolution and their putative role in sperm transport and selection within the uterus and the ability to fertilize the oocyte.

  19. A test of the sexy-sperm and good-sperm hypotheses for the evolution of polyandry

    OpenAIRE

    Kathryn B. McNamara; Emile van Lieshout; Leigh W. Simmons

    2014-01-01

    The sexy-sperm hypothesis proposes that polyandrous females will have a selective advantage over monandrous females as their sons will be sired by males with competitively superior sperm and will inherit this trait. Although the good-sperm hypothesis also predicts that offspring will be sired by males with competitively superior sperm, it additionally assumes a positive correlation between offspring sperm quality and the general viability of both sons and daughters. We examined the potential ...

  20. ß-Lysine discrimination by lysyl-tRNA synthetase

    DEFF Research Database (Denmark)

    Gilreath, Marla S; Roy, Hervé; Bullwinkle, Tammy J

    2011-01-01

    guided by the PoxA structure. A233S LysRS behaved as wild type with a-lysine, while the G469A and A233S/G469A variants decreased stable a-lysyl-adenylate formation. A233S LysRS recognized ß-lysine better than wildtype, suggesting a role for this residue in discriminating a- and ß-amino acids. Both...

  1. Arginine and Lysine Transporters Are Essential for Trypanosoma brucei.

    Science.gov (United States)

    Mathieu, Christoph; Macêdo, Juan P; Hürlimann, Daniel; Wirdnam, Corina; Haindrich, Alexander C; Suter Grotemeyer, Marianne; González-Salgado, Amaia; Schmidt, Remo S; Inbar, Ehud; Mäser, Pascal; Bütikofer, Peter; Zilberstein, Dan; Rentsch, Doris

    2017-01-01

    For Trypanosoma brucei arginine and lysine are essential amino acids and therefore have to be imported from the host. Heterologous expression in Saccharomyces cerevisiae mutants identified cationic amino acid transporters among members of the T. brucei AAAP (amino acid/auxin permease) family. TbAAT5-3 showed high affinity arginine uptake (Km 3.6 ± 0.4 μM) and high selectivity for L-arginine. L-arginine transport was reduced by a 10-times excess of L-arginine, homo-arginine, canavanine or arginine-β-naphthylamide, while lysine was inhibitory only at 100-times excess, and histidine or ornithine did not reduce arginine uptake rates significantly. TbAAT16-1 is a high affinity (Km 4.3 ± 0.5 μM) and highly selective L-lysine transporter and of the compounds tested, only L-lysine and thialysine were competing for L-lysine uptake. TbAAT5-3 and TbAAT16-1 are expressed in both procyclic and bloodstream form T. brucei and cMyc-tagged proteins indicate localization at the plasma membrane. RNAi-mediated down-regulation of TbAAT5 and TbAAT16 in bloodstream form trypanosomes resulted in growth arrest, demonstrating that TbAAT5-mediated arginine and TbAAT16-mediated lysine transport are essential for T. brucei. Growth of induced RNAi lines could partially be rescued by supplementing a surplus of arginine or lysine, respectively, while addition of both amino acids was less efficient. Single and double RNAi lines indicate that additional low affinity uptake systems for arginine and lysine are present in T. brucei.

  2. Studies on lysine production by Bacillus megaterium | Ekwealor ...

    African Journals Online (AJOL)

    A Lysine-producing strain recovered from soil was found to produce large amount of the amino acid. The bacterium identified as Bacillus megaterium SP 14 accumulated a lysine yield of 3.56 mg/ml in a broth culture in 96 h. Fermentation experiments show that 8.0% (w/v) glucose and 4.0% (w/v) ammonium chloride used as ...

  3. Intracellular pH in sperm physiology.

    Science.gov (United States)

    Nishigaki, Takuya; José, Omar; González-Cota, Ana Laura; Romero, Francisco; Treviño, Claudia L; Darszon, Alberto

    2014-08-01

    Intracellular pH (pHi) regulation is essential for cell function. Notably, several unique sperm ion transporters and enzymes whose elimination causes infertility are either pHi dependent or somehow related to pHi regulation. Amongst them are: CatSper, a Ca(2+) channel; Slo3, a K(+) channel; the sperm-specific Na(+)/H(+) exchanger and the soluble adenylyl cyclase. It is thus clear that pHi regulation is of the utmost importance for sperm physiology. This review briefly summarizes the key components involved in pHi regulation, their characteristics and participation in fundamental sperm functions such as motility, maturation and the acrosome reaction. Copyright © 2014 Elsevier Inc. All rights reserved.

  4. Function of the sperm nuclear matrix.

    Science.gov (United States)

    Shaman, Jeffrey A; Yamauchi, Yasuhiro; Ward, W Steven

    2007-01-01

    Mammalian spermatozoa contain some of the most highly compact chromatin. This is due to the DNA binding proteins, the protamines, which replace most of the histones during spermiogenesis. This chromatin, however, shares some features with somatic cell chromatin. One of these is the organization of DNA into loop domains attached at their bases to a proteinaceous nuclear matrix. Several groups have shown that the sites at which DNA associates with the sperm nuclear matrix contain chromatin structures that are linked with specific functions. Recent data also suggest that the sperm nuclear matrix plays essential roles in the paternal pronucleus of the newly fertilized oocyte, suggesting that the sperm cell provides more information to the new embryo than solely the genetic material it delivers. Here, we will review these data which together give insight into the functional significance and requirements of sperm nuclear structure.

  5. Life after Death. Sperm, Property, and Hope

    DEFF Research Database (Denmark)

    Kroløkke, Charlotte; Willum Adrian, Stine

    A tragic workplace accident led in 2010 to the death of Mark Edwards. Subsequently, his wife Joycelyn Edwards requested for the New South Wales Superior Court´s permission to extract sperm posthumous from her husband. She got the permission and one year later, the Court decided to grant her access...... Danish legal and bioethical discussions on the use of dead men’s sperm, and similar debates in New South Wales that led to the Edwards court ruling....... to the sperm her husband had deposited. Following a vivid media debate has taken place while the legislation was processed in the Parliament changing the legislation by legalizing the use of sperm from a dead man. Freezing technologies and extraction techniques has with the development of new reproductive...

  6. Mammalian Sperm Motility: Observation and Theory

    KAUST Repository

    Gaffney, E.A.

    2011-01-21

    Mammalian spermatozoa motility is a subject of growing importance because of rising human infertility and the possibility of improving animal breeding. We highlight opportunities for fluid and continuum dynamics to provide novel insights concerning the mechanics of these specialized cells, especially during their remarkable journey to the egg. The biological structure of the motile sperm appendage, the flagellum, is described and placed in the context of the mechanics underlying the migration of mammalian sperm through the numerous environments of the female reproductive tract. This process demands certain specific changes to flagellar movement and motility for which further mechanical insight would be valuable, although this requires improved modeling capabilities, particularly to increase our understanding of sperm progression in vivo. We summarize current theoretical studies, highlighting the synergistic combination of imaging and theory in exploring sperm motility, and discuss the challenges for future observational and theoretical studies in understanding the underlying mechanics. © 2011 by Annual Reviews. All rights reserved.

  7. Rat sperm motility analysis: methodologic considerations

    Science.gov (United States)

    The objective of these studies was to optimize conditions for computer-assisted sperm analysis (CASA) of rat epididymal spermatozoa. Methodologic issues addressed include sample collection technique, sampling region within the epididymis, type of diluent medium used, and sample c...

  8. Is lithium essential for epididymal sperm maturation?

    Science.gov (United States)

    Halder, Tanmoy; Datta, Uttam; Basu, Siddhartha; Mukherjee, Prasenjit

    2016-11-01

    A wider biological role of ultratrace element lithium in the mammalian reproduction has been reported, however, presence of lithium in the epididymal luminal fluid (ELF) and its influence on sperm during maturation events in the epididymal regions are still unknown. A pilot study was carried out in Jamunapari buck which revealed that levels of lithium in the ELF diminished gradually and significantly (Psperm were observed, except spermatozoan motility that was found absent in the caput epididymis. Therefore, we hypothesize that levels of lithium in the epididymal regions is one of the motility initiation and/or regulatory factor for epididymal sperm maturation essential for acquiring fertilizing competence of sperm cells, hence, lithium could also be considered as one of the biomarker of sperm maturation in any species. Copyright © 2016 Elsevier Ltd. All rights reserved.

  9. Biofortification of rice with lysine using endogenous histones.

    Science.gov (United States)

    Wong, H W; Liu, Q; Sun, S S M

    2015-02-01

    Rice is the most consumed cereal grain in the world, but deficient in the essential amino acid lysine. Therefore, people in developing countries with limited food diversity who rely on rice as their major food source may suffer from malnutrition. Biofortification of stable crops by genetic engineering provides a fast and sustainable method to solve this problem. In this study, two endogenous rice lysine-rich histone proteins, RLRH1 and RLRH2, were over-expressed in rice seeds to achieve lysine biofortification. Their protein sequences passed an allergic sequence-based homology test. Their accumulations in rice seeds were raised to a moderate level by the use of a modified rice glutelin 1 promoter with lowered expression strength to avoid the occurrence of physiological abnormalities like unfolded protein response. The expressed proteins were further targeted to protein storage vacuoles for stable storage using a glutelin 1 signal peptide. The lysine content in the transgenic rice seeds was enhanced by up to 35 %, while other essential amino acids remained balanced, meeting the nutritional standards of the World Health Organization. No obvious unfolded protein response was detected. Different degrees of chalkiness, however, were detected in the transgenic seeds, and were positively correlated with both the levels of accumulated protein and lysine enhancement. This study offered a solution to the lysine deficiency in rice, while at the same time addressing concerns about food safety and physiological abnormalities in biofortified crops.

  10. The sperm epigenome: implications for the embryo.

    Science.gov (United States)

    Gannon, John R; Emery, Benjamin R; Jenkins, Timothy G; Carrell, Douglas T

    2014-01-01

    Recent advances, including the human genome project and numerous studies of cancer and other diseases, have shown that the genetic code is not simply limited to the sequence of the four bases of DNA but also includes epigenetic programming, heritable changes that affect gene expression [Riggs A, Martinssen R, Russo V (2007) Introduction. In: Riggs A, Martinssen R, Russo V (eds) Epigenetics mechanisms of gene regulation. Cold Spring Harbor Press, New York]. The science of epigenetics is important in understanding many diseases and biological processes, including in identifying the causes of disease and better understanding the mechanisms by which the environment can affect gene expression [Carrell Fertil Steril 97 (2):267-274, 2012]. This chapter will focus on the epigenome of sperm and particularly highlight the potential role of the sperm epigenome in embryogenesis.The sperm epigenome is unique and highly specialized because of the unique nature and function of sperm and because of the diverse requirements for successful fertilization. Due to the need for motility, sperm chromatin must be compacted and highly organized. During spermiogenesis the chromatin is packaged tightly into the sperm head by the replacement of most histones with protamines. This allows for protection of the DNA from the hostile environment in the female reproductive tract. Remaining histones can have chemical modifications to the tails of the protein that either facilitate or repress gene transcription. Sperm, like embryonic stem cells, have a unique pattern of histone modifications that includes both activating and silencing marks in the promoters of genes associated with development. These bivalent marks, along with DNA hypomethylation, comprise a unique state in which the key genes are "poised" for possible activation in embryogenesis. Sperm epigenetic abnormalities have been linked with multiple diseases including male factor infertility and poor embryogenesis.

  11. A beta-lysine adenylating enzyme and a beta-lysine binding protein involved in poly beta-lysine chain assembly in nourseothricin synthesis in Streptomyces noursei.

    Science.gov (United States)

    Grammel, Nicolas; Pankevych, Kvitka; Demydchuk, Julia; Lambrecht, Klaus; Saluz, Hans-Peter; Krügel, Hans

    2002-01-01

    Nourseothricins (syn. Streptothricins), a group of nucleoside peptides produced by several streptomycete strains, contain a poly beta-lysine chain of variable length attached in amide linkage to the amino sugar moiety gulosamine of the nucleoside portion. We show that the nourseothricin-producing Streptomyces noursei contains an enzyme (NpsA) of an apparent M(r) 56,000 that specifically activates beta-lysine by adenylation but does not bind to it as a thioester. Cloning and sequencing of npsA from S. noursei including its flanking DNA regions revealed that it is closely linked to the nourseothricin resistance gene nat1 and some other genes on the chromosome possibly involved in nourseothricin biosynthesis. The deduced amino-acid sequence revealed that NpsA is a stand-alone adenylation domain with similarity to the adenylation domains of nonribosomal peptide synthetases (NRPS). Further analysis revealed that S. noursei contains a beta-lysine binding enzyme (NpsB) of about M(r) 64,100 which can be loaded by NpsA with beta-lysine as a thioester. Analysis of the deduced amino-acid sequence from the gene (npsB) of NpsB showed that it consists of two domains. The N-terminal domain of approximately 100 amino-acid residues has high similarity to PCP domains of NRPSs whereas the 450-amino-acid C-terminal domain has a high similarity to epimerization (E)-domains of NRPSs. Remarkably, in this E-domain the conserved H-H-motif is changed to H-Q, which suggests that either the domain is nonfunctional or has a specialized function. The presence of one single adenylating beta-lysine activating enzyme in nourseothricin-producing streptomycete and a separate binding protein suggests an iteratively operating NRPS-module catalyses synthesis of the poly beta-lysine chain.

  12. Seasonal changes in reproductive activity, sperm variables and sperm freezability in Blanca Andaluza bucks

    Energy Technology Data Exchange (ETDEWEB)

    Gallego-Calvo, L.; Gatica, M.C.; Santiago-Moreno, J.; Guzmán, J.L.; Zarazaga, L.

    2015-07-01

    Interest in the preservation of endangered breeds such as the Blanca Andaluza goat, has increased and some steps should be therefore taken to ensure it. The study was designed to determine the seasonal reproductive pattern of Blanca Andaluza bucks, and whether this affects the quality of their semen and its freezability over the year. Seven bucks were used and their body weight, testicular weight, plasma testosterone concentration and fresh sperm quality determined every week. The collected sperm was cryopreserved and stored; it was then thawed and the same sperm quality variables measured every fortnight. High plasma testosterone concentrations were recorded during the summer and autumn, and low concentrations were recorded during winter and spring (p<0.001). No differences were seen between seasons in terms of the percentage of bucks ejaculating, the percentage of active bucks, or ejaculate volume. However, the sperm concentration, the total number of sperm per ejaculate, and the values for most fresh sperm variables were lower during the winter period (at least p<0.05). After freezing-thawing, the quality of winter-collected sperm was better, in some respects, than that of summer-collected sperm (at least p<0.05). These results reveal that Blanca Andaluza bucks show seasonal reproductive activity in terms of their plasma testosterone concentration, but no clear change in their sexual behaviour between seasons was observed. The values of fresh sperm variables also vary over the year, reaching their lowest during winter. However, after freezing-thawing, winter-collected sperm is of overall better quality than sperm collected during the summer. (Author)

  13. Seasonal changes in reproductive activity, sperm variables and sperm freezability in Blanca Andaluza bucks

    Directory of Open Access Journals (Sweden)

    Lourdes Gallego-Calvo

    2015-12-01

    Full Text Available Interest in the preservation of endangered breeds such as the Blanca Andaluza goat, has increased and some steps should be therefore taken to ensure it. The study was designed to determine the seasonal reproductive pattern of Blanca Andaluza bucks, and whether this affects the quality of their semen and its freezability over the year. Seven bucks were used and their body weight, testicular weight, plasma testosterone concentration and fresh sperm quality determined every week. The collected sperm was cryopreserved and stored; it was then thawed and the same sperm quality variables measured every fortnight. High plasma testosterone concentrations were recorded during the summer and autumn, and low concentrations were recorded during winter and spring (p<0.001. No differences were seen between seasons in terms of the percentage of bucks ejaculating, the percentage of active bucks, or ejaculate volume. However, the sperm concentration, the total number of sperm per ejaculate, and the values for most fresh sperm variables were lower during the winter period (at least p<0.05. After freezing-thawing, the quality of winter-collected sperm was better, in some respects, than that of summer-collected sperm (at least p<0.05. These results reveal that Blanca Andaluza bucks show seasonal reproductive activity in terms of their plasma testosterone concentration, but no clear change in their sexual behaviour between seasons was observed. The values of fresh sperm variables also vary over the year, reaching their lowest during winter. However, after freezing-thawing, winter-collected sperm is of overall better quality than sperm collected during the summer.

  14. Oxidative stress in zebrafish (Danio rerio sperm.

    Directory of Open Access Journals (Sweden)

    Mary Hagedorn

    Full Text Available Laboratories around the world have produced tens of thousands of mutant and transgenic zebrafish lines. As with mice, maintaining all of these valuable zebrafish genotypes is expensive, risky, and beyond the capacity of even the largest stock centers. Because reducing oxidative stress has become an important aspect of reducing the variability in mouse sperm cryopreservation, we examined whether antioxidants might improve cryopreservation of zebrafish sperm. Four experiments were conducted in this study. First, we used the xanthine-xanthine oxidase (X-XO system to generate reactive oxygen species (ROS. The X-XO system was capable of producing a stress reaction in zebrafish sperm reducing its sperm motility in a concentration dependent manner (P0.05, whereas superoxide dismutase (SOD and vitamin E did not (P0.05, producing a mean 2.0 to 2.9-fold increase in post-thaw motility. Fourth, we examined the effect of additional cryoprotectants and CAT on fresh sperm motility. Cryoprotectants, 8% methanol and 10% dimethylacetamide (DMA, reduced the motility over the control value (P0.05. Zebrafish sperm protocols should be modified to improve the reliability of the cryopreservation process, perhaps using a different cryoprotectant. Regardless, the simple addition of CAT to present-day procedures will significantly improve this process, assuring increased and less variable fertilization success and allowing resource managers to dependably plan how many straws are needed to safely cryopreserve a genetic line.

  15. Viscoelasticity promotes collective swimming of sperm

    Science.gov (United States)

    Tung, Chih-Kuan; Harvey, Benedict B.; Fiore, Alyssa G.; Ardon, Florencia; Suarez, Susan S.; Wu, Mingming

    From flocking birds to swarming insects, interactions of organisms large and small lead to the emergence of collective dynamics. Here, we report striking collective swimming of bovine sperm, with sperm orienting in the same direction within each cluster, enabled by the viscoelasticity of the fluid. A long-chain polyacrylamide solution was used as a model viscoelastic fluid such that its rheology can be fine-tuned to mimic that of bovine cervical mucus. In viscoelastic fluid, sperm formed dynamic clusters, and the cluster size increased with elasticity of the polyacrylamide solution. In contrast, sperm swam randomly and individually in Newtonian fluids of similar viscosity. Analysis of the fluid motion surrounding individual swimming sperm indicated that sperm-fluid interaction is facilitated by the elastic component of the fluid. We note that almost all biological fluids (e.g. mucus and blood) are viscoelastic in nature, this finding highlights the importance of fluid elasticity in biological function. We will discuss what the orientation fluctuation within a cluster reveals about the interaction strength. Supported by NIH Grant 1R01HD070038.

  16. Oxidative stress negatively affects human sperm mitochondrial respiration.

    Science.gov (United States)

    Ferramosca, Alessandra; Pinto Provenzano, Sara; Montagna, Daniela Domenica; Coppola, Lamberto; Zara, Vincenzo

    2013-07-01

    To correlate the level of oxidative stress in serum and seminal fluid and the level of sperm deoxyribonucleic acid (DNA) fragmentation with sperm mitochondrial respiratory efficiency. Sperm mitochondrial respiratory activity was evaluated with a polarographic assay of oxygen consumption carried out in hypotonically treated sperm cells. A possible relationship between sperm mitochondrial respiratory efficiency, the level of oxidative stress, and the level of sperm DNA fragmentation was investigated. Sperm motility was positively correlated with mitochondrial respiration but negatively correlated with oxidative stress and DNA fragmentation. Interestingly, sperm mitochondrial respiratory activity was negatively affected by oxidative stress and DNA fragmentation. Our data indicate that sperm mitochondrial respiration is decreased in patients with high levels of reactive oxygen species by an uncoupling between electron transport and adenosine triphosphate synthesis. This reduction in mitochondrial functionality might be 1 of the reasons responsible for the decrease in spermatozoa motility. Copyright © 2013 Elsevier Inc. All rights reserved.

  17. Physiological performance of juvenile Haliotis rufescens and Haliotis discus hannai abalone exposed to the withering syndrome agent.

    Science.gov (United States)

    González, Roxana C; Brokordt, Katherina; Lohrmann, Karin B

    2012-09-15

    Withering syndrome (WS) is a serious chronic disease caused by infection with the bacterium Candidatus Xenohaliotis californiensis, a Rickettsiales-like organism (WS-RLO) that affects multiple abalone species in both natural and farmed populations. However, there is no available information regarding the effects of this disease on the physiological performance of infected abalone. We studied the effect of different levels of infection on components of energy balance and physiological indices (rates of absorption and assimilation, O/N ratio, and scope for growth) in the abalone species Haliotis rufescens and Haliotis discus hannai. Juveniles were exposed to C. X. californiensis transmission for 130 days, during which time the presence/absence of WS-RLOs was evaluated by PCR (following DNA sequencing-based confirmation of 100% identity with the sequence of C. X. californiensis from California), and the prevalence and intensity of infection were evaluated via histological analysis. Among H. rufescens juveniles exposed to the bacterium, 92% became infected (positive by histology), and the intensity of infection ranged from low (degree 1) to moderate (degree 2). In contrast, no H. discus hannai juveniles were positive for WS-RLO by histology, although 23% were positive by PCR, possibly indicating incipient WS-RLO infection that did not develop during the experimental period or to mere presence of WS-RLO DNA in the sample. Infection of H. rufescens juveniles by WS-RLOs negatively affected all components of the energy balance and physiological indices, such as scope for growth and the O/N ratio, in direct relation to the degree of infection. The most strongly affected functions were the rate of ingestion, standard metabolism, and production of feces, which were reduced by 60-80% in the most highly infected individuals. The reduced energy intake in the organisms produced a strong energy imbalance such that the energy available for growth was reduced by 49% in infected

  18. Polaribacter haliotis sp. nov., isolated from the gut of abalone Haliotis discus hannai.

    Science.gov (United States)

    Kim, Young-Ok; Park, In-Suk; Park, Sooyeon; Nam, Bo-Hye; Park, Ji-Min; Kim, Dong-Gyun; Yoon, Jung-Hoon

    2016-12-01

    A Gram-stain-negative, aerobic, non-motile and coccoid, ovoid or rod-shaped bacterial strain, designated RA4-7T, was isolated from the gut of an abalone (Haliotis discus hannai) collected from the sea around Jeju island, South Korea, and subjected to a polyphasic taxonomic study. RA4-7T grew optimally at 25 °C, at pH 7.0-7.5 and in the presence of 2.0-3.0 % (w/v) NaCl. The phylogenetic trees based on 16S rRNA gene sequences showed that RA4-7T represented a member of the genus Polaribacter. RA4-7T exhibited 16S rRNA gene sequence similarity values of 97.64 and 97.23 % to Polaribacter atrinae WP25T and Polaribacter dokdonensis DSW-5T, respectively, and of 93.83-96.99 % to the type strains of the other species of the genus Polaribacter. RA4-7T contained MK-6 as the predominant menaquinone and summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c), iso-C15 : 0 3-OH and iso-C15 : 0as the major fatty acids. The major polar lipids detected in RA4-7T were phosphatidylethanolamine and one unidentified lipid. The DNA G+C content of RA4-7T was 30.5 mol% and its DNA-DNA relatedness values with the type strains of P. atrinae and P. dokdonensis were 16 and 11 %, respectively. Differential phenotypic properties, in combination with its phylogenetic and genetic distinctiveness, revealed that RA4-7T is separated from species of the genus Polaribacter with validly published names. On the basis of the data presented, RA4-7T represented a novel species of the genus Polaribacter, for which the name Polaribacterhaliotis sp. nov. is proposed. The type strain is RA4-7T (=KCTC 52418T=NBRC 112383T).

  19. Effects of ocean acidification with pCO2diurnal fluctuations on survival and larval shell formation of Ezo abalone, Haliotis discus hannai.

    Science.gov (United States)

    Onitsuka, Toshihiro; Takami, Hideki; Muraoka, Daisuke; Matsumoto, Yukio; Nakatsubo, Ayumi; Kimura, Ryo; Ono, Tsuneo; Nojiri, Yukihiro

    2018-03-01

    This study assessed the effects of constant and diurnally fluctuating pCO 2 on development and shell formation of larval abalone Haliotis discus hannai. The larvae was exposed to different pCO 2 conditions; constant [450, 800, or 1200 μatm in the first experiment (Exp. I), 450 or 780 μatm in the second experiment (Exp. II)] or diurnally fluctuating pCO 2 (800 ± 400 or 1200 ± 400 μatm in Exp. I, 450 ± 80, 780 ± 200 or 780 ± 400 μatm in Exp. II). Mortality, malformation rates or shell length of larval abalone were not significantly different among the 450, 800, and 800 ± 400 μatm pCO 2 treatments. Meanwhile, significantly higher malformation rates and smaller shells were detected in the 1200 and 1200 ± 400 μatm pCO 2 treatments than in the 450 μatm pCO 2 treatment. The negative impacts were greater in the 1200 ± 400 μatm than in the 1200 μatm. Shell length and malformation rate of larval abalone were related with aragonite saturation state (Ω-aragonite) in experimental seawater, and greatly changed around 1.1 of Ω-aragonite which corresponded to 1000-1300 μatm pCO 2 . These results indicate that there is a pCO 2 threshold associated with Ω-aragonite in the seawater, and that pCO 2 fluctuations produce additional negative impacts on abalone when above the threshold. Clear relationships were detected between abalone fitness and the integrated pCO 2 value over the threshold, indicating that the effects of OA on development and shell formation of larval abalone can be determined by intensity and time of exposure to pCO 2 over the threshold. Copyright © 2017 Elsevier Ltd. All rights reserved.

  20. A novel potentiometric biosensor for determination of L-lysine in commercial pharmaceutical L-lysine tablet and capsule.

    Science.gov (United States)

    Yarar, Saniye; Karakuş, Emine

    2016-01-01

    The construction of an L-lysine biosensor on ammonium-selective poly(vinylchloride) (PVC) membrane electrode is described in this study. The construction procedure occurs in two stages: (I) the preparation of ammonium-selective poly(vinylchloride) (PVC) membrane electrode and (II) the chemical immobilization of lysine oxidase on this ammonium-selective electrode by using glutaraldehyde. The ammonium ions produced after enzymatic reaction were determined potentiometrically. The sensitivity of the lysine biosensor against ammonium ions and lysine were studied. The response time, linear working range, reproducibility and life time of the biosensor were also determined. The interfering effect of other amino acids on the biosensor performance was also studied and potentiometric selectivity coefficients were calculated. Although the biosensor responded mainly against tyrosine, a lot of amino acids and ascorbic acid that can be present in some real samples did not show any important interference. Additionally, lysine assay in commercial pharmaceutical lysine tablets and capsules was also successfully carried out. The results were in good agreement with previously reported values.

  1. Phenotypic plasticity in sperm traits in scorpionflies (Mecoptera : Panorpidae): Consequences of larval history and seasonality on sperm length and sperm transfer

    NARCIS (Netherlands)

    Vermeulen, Andreas; Engels, Sierk; Engqvist, Leif; Sauer, Klaus Peter

    2009-01-01

    We examined effects of seasonality, larval food availability and larval rearing density on sperm length, sperm transfer rates and body size in the bivoltine scorpionfly Panorpa vulgaris. Males of the first annual generation were larger and had larger sperm. Comparing individuals of two summer

  2. Sexual selection and the evolution of sperm quality.

    Science.gov (United States)

    Fitzpatrick, John L; Lüpold, Stefan

    2014-12-01

    Sperm experience intense and varied selection that dramatically impacts the evolution of sperm quality. Selection acts to ensure that sperm are fertilization-competent and able to overcome the many challenges experienced on their way towards eggs. However, simply being able to fertilize an egg is not enough to ensure male fertility in most species. Owing to the prevalence of female multiple mating throughout the animal kingdom, successful fertilization requires sperm to outcompete rival sperm. In addition, females can actively influence sperm quality, storage or utilization to influence male fertility. This review provides an overview of how these selective forces influence the evolution of sperm quality. After exploring the link between sperm traits and male fertility, we examine how post-mating competition between rival ejaculates influences the evolution of sperm quality. We then describe how complex genetic, social and sexual interactions influence sperm quality, focusing on the importance of seminal fluid and interactions between sperm and the female's reproductive tract. In light of the complexities of selection on sperm traits, greater use of multivariate approaches that incorporate male-male, sperm-sperm and sperm-female interactions to study sperm quality will enhance our understanding of how selection acts on sperm traits and factors influencing male fertility. Because the metric of male reproductive success--fertilization--is the same across the animal kingdom, we argue that information about sperm evolution gained from non-human animals has enormous potential to further our understanding of the factors that impact human fertility. © The Author 2014. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  3. Variation in sperm morphometry and sperm competition among barn swallow (Hirundo rustica) populations

    Czech Academy of Sciences Publication Activity Database

    Laskemoen, T.; Albrecht, Tomáš; Bonisoli-Alquati, A.; Cepák, J.; De Lope, F.; Hermosell, I. G.; Johannessen, L. E.; Kleven, O.; Marzal, A.; Mousseau, T. A.; Moller, A. P.; Robertson, R. J.; Rudolfsen, G.; Saino, N.; Vortman, Y.; Lifjeld, J. T.

    2013-01-01

    Roč. 67, č. 2 (2013), s. 301-309 ISSN 0340-5443 R&D Projects: GA ČR(CZ) GAP506/12/2472 Institutional support: RVO:68081766 Keywords : Barn swallow * Extrapair paternity * Hirundo rustica * Sperm competition * Sperm size Subject RIV: EG - Zoology Impact factor: 3.049, year: 2013

  4. Toward microfluidic sperm refinement: continuous flow label-free analysis and sorting of sperm cells

    NARCIS (Netherlands)

    de Wagenaar, B.; Dekker, Stefan; van den Berg, Albert; Segerink, Loes Irene

    2015-01-01

    This manuscript reports upon the development of a microfluidic setup to detect and sort sperm cells from polystyrene beads label-free and non-invasively. Detection is performed by impedance analysis. When sperm cells passed the microelectrodes, the recorded impedance (19.6 ± 5.7 Ω) was higher

  5. Sperm depletion as a consequence of increased sperm competition risk in the European bitterling, Rhodeus amarus

    Czech Academy of Sciences Publication Activity Database

    Smith, C.; Pateman-Jones, C.; Zieba, G.; Przybylski, M.; Reichard, Martin

    2009-01-01

    Roč. 77, č. 5 (2009), s. 1227-1233 ISSN 0003-3472 Institutional research plan: CEZ:AV0Z60930519 Keywords : alternative mating tactics * European bitterling * mating system * sexual conflict * sexual selection * sneaking * sperm competition * sperm economy Subject RIV: EG - Zoology Impact factor: 2.890, year: 2009

  6. Accountings of Selecting Sperm on the (ethical) border

    DEFF Research Database (Denmark)

    Willum Adrian, Stine

    During the past years, Denmark has become a destination for fertility travellers, in need for donated sperm. Today, treatment is possible no matter of marital status or sexuality. Furthermore, users of sperm donation can choose between anonymous and non-anonymous sperm, with either basic informat......During the past years, Denmark has become a destination for fertility travellers, in need for donated sperm. Today, treatment is possible no matter of marital status or sexuality. Furthermore, users of sperm donation can choose between anonymous and non-anonymous sperm, with either basic...

  7. What use is an infertile sperm? A comparative study of sperm-heteromorphic Drosophila

    DEFF Research Database (Denmark)

    Holman, Luke; Freckleton, Robert P; Snook, Rhonda R

    2007-01-01

    Sperm size and number are important determinants of male reproductive success. The genus Drosophila exhibits a remarkable diversity of sperm production strategies, including the production of multiple sperm morphs by individual males, a phenomenon called sperm heteromorphism. Sperm......-heteromorphic Drosophila species in the obscura group produce large numbers of infertile "parasperm" in addition to fertile eusperm. Parasperm have been hypothesized to perform a number of roles in place of fertilization, predominantly focused on their potential function in postcopulatory sexual selection. However......, the evolutionary significance of parasperm remains unknown. Here we measured several male and female morphological, behavioral, and life-history traits in 13 obscura group species to test competing hypotheses of parasperm function using comparative methods. We found that parasperm size was unrelated to female...

  8. iTRAQ-Based Identification of Proteins Related to Muscle Growth in the Pacific Abalone, Haliotis discus hannai.

    Science.gov (United States)

    Huang, Jianfang; You, Weiwei; Luo, Xuan; Ke, Caihuan

    2017-10-25

    The abalone Haliotis discus hannai is an important aquaculture species that is grown for human consumption. However, little is known of the genetic mechanisms governing muscle growth in this species, particularly with respect to proteomics. The isobaric tag for relative and absolute quantitation (iTRAQ) method allows for sensitive and accurate protein quantification. Our study was the first to use iTRAQ-based quantitative proteomics to investigate muscle growth regulation in H. discus hannai . Among the 1904 proteins identified from six samples, 125 proteins were differentially expressed in large specimens of H. discus hannai as compared to small specimens. In the large specimens, 47 proteins were upregulated and 78 were downregulated. Many of the significant Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, including these differentially expressed proteins, were closely related to muscle growth, including apoptosis, thyroid hormone signaling, regulation of the actin cytoskeleton, and viral myocarditis ( p discus hannai .

  9. Thyroxin Is Useful to Improve Sperm Motility

    Directory of Open Access Journals (Sweden)

    Mendeluk Gabriela Ruth

    2016-07-01

    Full Text Available Background The aim of this study was to evaluate the non-genomic action of thyroxin on sperm kinetic and its probable use to improve sperm recovery after applying an en- richment method like “swim-up” in comparison with the available one, pentoxifylline. Materials and Methods This is an experimental study. A total of 50 patients were re- cruited, followed by infertility consultation. Conventional sperm assays were performed according to World Health Organization criteria-2010 (WHO-2010. A Computer Aided Semen Analysis System was employed to assess kinetic parameters and concentrations. Number of the motile sperm recovered after preparation technique was calculated. Results Addition of T4 (0.002 µg/ml to semen samples increased hypermotility at 20 minutes (control: 14.18 ± 5.1% vs. 17.66 ± 8.88%, P<0.03, data expressed as mean ± SD and remained unchanged after 40 minutes. Significant differences were found in the motile sperm recovered after swim-up (control: 8.93×106 ± 9.52× 06vs. 17.20×106 ± 21.16×106, P<0.03, achieving all of the tested samples a desirable threshold value for artificial insemination outcome, while adding pentoxifylline increased the number of recovered sperm after swim-up in 60% of the studied cases. No synergism between two treatments could be determined. Conclusion We propose a new physiological tool to artificially improve insemination. The discussion opens windows to investigate unknown pathways involved in sperm ca- pacitation and gives innovative arguments to better understand infertility mechanisms.

  10. Endocannabinoids and Human Sperm Cells

    Directory of Open Access Journals (Sweden)

    Giovanna Zolese

    2010-10-01

    Full Text Available N-acylethanolamides (NAEs are naturally occurring signaling lipids consisting of amides and esters of long-chain polyunsaturated fatty acids. Usually they are present in a very small amounts in many mammalian tissues and cells, including human reproductive tracts and fluids. Recently, the presence of N-arachidonoylethanolamide (anandamide, AEA, the most characterised member of endocannabinoids, and its congeners palmitoylethanolamide (PEA and oleylethanolamide (OEA in seminal plasma, oviductal fluid, and follicular fluids was demonstrated. AEA has been shown to bind not only type-1 (CB1 and type-2 (CB2 cannabinoid receptors, but also type-1 vanilloid receptor (TRPV1, while PEA and OEA are inactive with respect to classical cannabinoid CB1 and CB2 but activate TRPV1 or peroxisome proliferator activate receptors (PPARs. This review concerns the most recent experimental data on PEA and OEA, endocannabinoid-like molecules which appear to exert their action exclusively on sperm cells with altered features, such as membrane characteristics and kinematic parameters. Their beneficial effects on these cells could suggest a possible pharmacological use of PEA and OEA on patients affected by some forms of idiopathic infertility.

  11. Marine reserves help preserve genetic diversity after impacts derived from climate variability: Lessons from the pink abalone in Baja California

    Directory of Open Access Journals (Sweden)

    Adrián Munguía-Vega

    2015-07-01

    Full Text Available Genetic diversity is crucial for the adaptation of exploited species like the pink abalone (Haliotis corrugata, faced with threats from climate change, overfishing and impacts associated with aquaculture production. While marine reserves are commonly used to mitigate risks to marine populations, the duration, size, location and larval connectivity needed for a reserve to help conserve genetic resources is still poorly understood. Here, we examine the effects of fishing, reserves, and restocking on the genetic diversity of 10 populations from central Baja California, Mexico, and Southern California, USA. We demonstrate that each population shows characteristic genetic signatures according to recent management decisions. We found high allelic diversity, particularly rare alleles, a larger effective population size and a lack of a recent genetic bottleneck in pink abalones within a small (0.8 km2, recently established (5 years reserve in Baja California, compared to other fished sites after a climatic bottleneck. Higher diversity may result from the presence of older animals in the reserve. Due to its location, the reserve may also act as an important hub connecting distant populations via larval dispersal. In contrast, a population from California showed genetic isolation, loss of allelic diversity and high relatedness, consistent with the collapse of fisheries in the 1990s and their lack of recovery thereafter. In addition, a fished area in Baja California with a history of restocking for over a decade showed an increase in frequency of related individuals and high genetic differentiation from nearby sites that were consistent with the production of larvae from a few adults in the laboratory. A network of strategically placed small marine reserves that considers ocean circulation patterns could help to maintain genetic diversity and connectivity of exploited populations.

  12. Kazal-type proteinase inhibitor from disk abalone (Haliotis discus discus): molecular characterization and transcriptional response upon immune stimulation.

    Science.gov (United States)

    Wickramaarachchi, W D Niroshana; De Zoysa, Mahanama; Whang, Ilson; Wan, Qiang; Lee, Jehee

    2013-09-01

    Proteinases and proteinase inhibitors are involved in several biological and physiological processes in all multicellular organisms. Proteinase inhibitors play a key role in regulating the activity of the respective proteinases. Among serine proteinase inhibitors, kazal-type proteinase inhibitors (KPIs) are widely found in mammals, avians, and a variety of invertebrates. In this study, we describe the identification of a kazal-type serine proteinase inhibitor (Ab-KPI) from the disk abalone, Haliotis discus discus, which is presumably involved in innate immunity. The full-length cDNA of Ab-KPI includes 600 bp nucleotides with an open reading frame (ORF) encoding a polypeptide of 143 amino acids. The deduced amino acid sequence of Ab-KPI contains a putative 17-amino acid signal peptide and two tandem kazal domains with high similarity to other kazal-type SPIs. Each kazal domain consists of reactive site (P1) residue containing a leucine (L), and a threonine (T) located in the second amino acid position after the second conserved cysteine of each domain. Temporal expression of Ab-KPI was assessed by real time quantitative PCR in hemocytes and mantle tissue following bacterial and viral hemorrhagic septicemia virus (VHSV) challenge, and tissue injury. At 6 h post-bacterial and -VHSV challenge, Ab-KPI expression in hemocytes was increased 14-fold and 4-fold, respectively, compared to control samples. The highest up-regulations upon tissue injury were shown at 9 h and 12 h in hemocytes and mantle, respectively. The transcriptional modulation of Ab-KPI following bacterial and viral challenges and tissue injury indicates that it might be involved in immune defense as well as wound healing process in abalone. Copyright © 2013 Elsevier Ltd. All rights reserved.

  13. Ferritin from the Pacific abalone Haliotis discus hannai: Analysis of cDNA sequence, expression, and activity.

    Science.gov (United States)

    Qiu, Reng; Kan, Yunchao; Li, Dandan

    2016-02-01

    Ferritin plays an important role in iron homeostasis due to its ability to bind and sequester large amounts of iron. In this study, the gene encoding a ferritin (HdhFer2) was cloned from Pacific abalone (Haliotis discus hannai). The full-length cDNA of HdhFer2 contains a 5'-UTR of 121 bp, an ORF of 516 bp, and a 3'-UTR of 252 bp with a polyadenylation signal sequence of AATAAA and a poly(A) tail. It also contains a 31 bp iron-responsive element (IRE) in the 5'-UTR position, which is conserved in many ferritins. HdhFer2 consists of 171 amino acid residues with a predicted molecular weight (MW) ∼19.8 kDa and a theoretical isoelectric point (PI) of 4.84. The deduced amino acid sequence of HdhFer2 contains two ferritin iron-binding region signatures (IBRSs). HdhFer2 mRNA was detected in a wide range of tissues and was dominantly expressed in the gill. Infection with the bacterial pathogen Vibrio anguillarum significantly upregulated HdhFer2 expression in a time-dependent manner. Recombinant HdhFer2 (rHdhFer2) purified from Escherichia coli was able to bind ferrous iron in a concentration-dependent manner. In summary, these results suggest that HdhFer2 is a crucial protein in the iron-withholding defense system, and plays an important role in the innate immune response of abalone. Copyright © 2016 Elsevier Ltd. All rights reserved.

  14. Protamine-like sperm nuclear basic proteins in the primitive frog Ascaphus truei and histone reversions among more advanced frogs.

    Science.gov (United States)

    Kasinsky, H E; Gutovich, L; Kulak, D; Mackay, M; Green, D M; Hunt, J; Ausio, J

    1999-12-01

    Sperm nuclear basic proteins (SNBPs) that condense chromatin are very diverse. In animals, evolution of SNBPs has proceeded from lysine-rich histone H type in sponges to more arginine-rich protamine-like PL and protamine P types. Yet sporadic PL/P to H reversions are known to occur in both protostomes and deuterostomes. To determine why this is the case, we have examined SNBPs in eleven anuran species. We find that sperm of the primitive, internally fertilizing archeobatrachian frog A. truei (family Ascaphidae) has PL/P type (42 mol % arginine), with an electrophoretic profile similar to SNBPs in another archeobatrachian, externally fertilizing Leiopelma hochstetteri (family Leiopelmatidae). Cytochemistry of sperm nuclei in the advanced, externally fertilizing neobatrachian frogs Crinia signifera and C. deserticola (family Myobatrachidae) indicates that they have reverted to H type SNBPs. This is also known to be the case in externally fertilizing Rana (family Ranidae) and Silurana, a subgenus of Xenopus (family Pipidae). Such a trend, from PL/P type SNBPs in two archeobatrachians to sporadic reversions to H type in more advanced frogs, parallels the ultrastructural simplification from complex A. truei introsperm to neobatrachian aquasperm that Jamieson et al. (1993. Herpetologica 49:52-65) attribute as a secondary reversion to external fertilization.

  15. Experimental evolution of sperm competitiveness in a mammal

    Directory of Open Access Journals (Sweden)

    Simmons Leigh W

    2011-01-01

    Full Text Available Abstract Background When females mate with multiple partners, sperm from rival males compete to fertilise the ova. Studies of experimental evolution have proven the selective action of sperm competition on male reproductive traits. However, while reproductive traits may evolve in response to sperm competition, this does not necessarily provide evidence that sperm competitive ability responds to selection. Indeed, a study of Drosophila failed to observe divergence in sperm competitive ability of males in lines selected for enhanced sperm offence and defence. Results Adopting the naturally polygamous house mouse (Mus domesticus as our vertebrate model, we performed an experimental evolution study and observed genetic divergence in sperm quality; males from the polygamous selection lines produced ejaculates with increased sperm numbers and greater sperm motility compared to males from the monogamous lines. Here, after 12 generations of experimental evolution, we conducted competitive matings between males from lineages evolving under sperm competition and males from lineages subject to relaxed selection. We reduced variation in paternity arising from embryo mortality by genotyping embryos in utero at 14 days gestation. Our microsatellite data revealed a significant paternity bias toward males that evolved under the selective regime of sperm competition. Conclusion We provide evidence that the sperm competitiveness phenotype can respond to selection, and show that improved sperm quality translates to greater competitive fertilisation success in house mice.

  16. Analysis of sperm quality in recombinant inbred mouse strains: correlation of sperm head shape with sperm abnormalities and with the incidence of supplementary spermatozoa in the perivitelline space.

    Science.gov (United States)

    Krzanowska, H; Styrna, J; Wabik-Sliz, B

    1995-07-01

    Recombinant inbred strains were developed from reciprocal crosses between two inbred strains of mice (CBA and KE) differing in sperm head shape, proportion of normal sperm heads (CBA, 95%; KE, 78%) and fertilization efficiency (CBA, 100% of fertilized ova; KE, 72%), to determine whether the indices of sperm morphology and function were correlated. The following parameters were analysed in recombinant inbred and progenitor strains: index of sperm head shape (head width in the middle of its length/head length), percentage of abnormal sperm heads, percentage of spermatozoa with progressive movements, efficiency of penetration of hyaluronic acid polymer (Sperm Select) and percentage of fertilized ova after mating males from the tested strains with females from an outbred stock. For each investigated character, recombinant inbred strains, recombinant inbred EXCB and CBXE, could be divided into at least three categories: KE-like, CBA-like and intermediate, suggesting that in each case a minimum of two genes was involved. Recombinant strains derived from the reciprocal crosses of progenitor strains differed only with respect to the proportion of abnormal sperm heads, showing the involvement of the Y chromosome in determining this character. Penetration into Sperm Select was significantly correlated both with fertilization efficiency and sperm motility, while correlation with the proportion of normal spermatozoa did not reach the level of significance. However, there was a significant negative correlation of both sperm abnormalities and the incidence of supplementary spermatozoa in the perivitelline space with the index of sperm head shape.(ABSTRACT TRUNCATED AT 250 WORDS)

  17. Nutritional consequences of interspecies differences in arginine and lysine metabolism.

    Science.gov (United States)

    Ball, Ronald O; Urschel, Kristine L; Pencharz, Paul B

    2007-06-01

    Differences in lysine and arginine requirements among various species such as omnivores (humans, pigs, rats, dogs), carnivores (cats), herbivores (rabbits, horses), ruminants (cattle), poultry, and fish, are covered in detail in this article. Although lysine is classified as an indispensable amino acid across species, the classification of arginine as either an indispensable or dispensable amino acid is more ambiguous because of differences among species in rates of de novo arginine synthesis. Because lysine is most often the limiting amino acid in the diet, its requirement has been extensively studied. By use of the ideal protein concept, the requirements of the other indispensable amino acids can be extrapolated from the lysine requirement. The successful use of this concept in pigs is compared with potential application of the ideal protein concept in humans. The current dietary arginine requirement varies widely among species, with ruminants, rabbits, and rats having relatively low requirements and carnivores, fish, and poultry having high requirements. Interspecies differences in metabolic arginine utilization and reasons for different rates of de novo arginine synthesis are reviewed in detail, as these are the primary determinants of the dietary arginine requirement. There is presently no dietary requirement for humans of any age, although this needs to be reassessed, particularly in neonates. A thorough understanding of the factors contributing to the lysine and arginine requirements in different species will be useful in our understanding of human amino acid requirements.

  18. Psychosocial counselling of identifiable sperm donors.

    Science.gov (United States)

    Visser, M; Mochtar, M H; de Melker, A A; van der Veen, F; Repping, S; Gerrits, T

    2016-05-01

    What do identifiable sperm donors feel about psychosocial counselling? Identifiable sperm donors found it important that psychosocial counselling focused on emotional consequences and on rules and regulations and they expected to have access to psychosocial counselling at the time that donor-offspring actually sought contact. Most studies on sperm donors are on anonymous donors and focus on recruitment, financial compensation, anonymity and motivations. There is limited knowledge on the value that identifiable sperm donors place on psychosocial counselling and what their needs are in this respect. We performed a qualitative study from March until June 2014 with 25 identifiable sperm donors, who were or had been a donor at the Centre for Reproductive Medicine of the Academic Medical Centre in Amsterdam any time between 1989 and 2014. We held semi-structured in-depth interviews with identifiable sperm donors with an average age of 44 years. The interviews were fully transcribed and analysed using the constant comparative method of grounded theory. Twelve out of 15 donors (former donors ITALIC! n = 8, active donors ITALIC! n = 7) who had received a counselling session during their intake procedure found it important that they had been able to talk about issues such as the emotional consequences of donation, disclosure to their own children, family and friends, future contact with donor-offspring and rules and regulations. Of the 10 former donors who had received no counselling session, 8 had regretted the lack of intensive counselling. In the years following their donation, most donors simply wanted to know how many offspring had been born using their sperm and had no need for further counselling. Nevertheless, they frequently mentioned that they were concerned about the well-being of 'their' offspring. In addition, they would value the availability of psychosocial counselling in the event that donor-offspring actually sought contact. A limitation of our study is its

  19. Molluskan fasciclin-1 domain-containing protein: molecular characterizationand gene expression analysis of fasciclin 1-like protein from disk abalone (Haliotis discus discus).

    Science.gov (United States)

    Premachandra, H K A; De Zoysa, Mahanama; Nikapitiya, Chamilani; Lee, Youngdeuk; Wickramaarachchi, W D N; Whang, Ilson; Lee, Jehee

    2013-06-15

    Cell-to-cell contacts play a key role in multicellular systems and organisms. Fasciclin-1 (FAS-1) is a lipid-linked membrane associated glycoprotein that is a member of a newly recognized family of cell adhesion molecules sharing features with the immunoglobulins, cadherins, integrins, and selectins. Here, we report the identification and molecular characterization of a novel FAS-1 domain-containing cDNA from disk abalone (Haliotis discus discus), including its gene expression profile and immune response to bacterial stimuli and tissue injuries. Designated as Abfac1, the 909bp open reading frame (ORF) encodes 303 amino acid (aa) residues with a predicted molecular mass of 33kDa and isoelectric (pI) value of 4.9. The aa sequence contains two FAS-1 domains and three conserved regions, FRa motif, H-box, and FRb motif. Phylogenetic analysis showed the closest relation to Jellyfish cell adhesion protein. In healthy abalone, Abfac1 expression is highest in hepatopancreas followed by mantle and lowest in digestive gland. In immune-stimulated abalones, relative Abfac1 mRNA expression was increased in hemocytes by ~11-fold at 48h after the Vibrio parahaemolyticus infection, by 3.1-fold at 6h after the Listeria monocytogenes infection and by ~9-fold at 6h after the LPS injection. Similarly, tissue injuries caused significant increase of relative mRNA expression by 3.5-fold in hemocytes and by ~10-fold in mantle at 12h post-injury. These results suggest that the novel member of the FAS-1 domain-containing protein family, Abfac1, may be involved in immune response and cell adhesion in disk abalone. Copyright © 2013 Elsevier B.V. All rights reserved.

  20. Structural and functional characterization of a novel molluskan ortholog of TRAF and TNF receptor-associated protein from disk abalone (Haliotis discus discus).

    Science.gov (United States)

    Lee, Youngdeuk; Elvitigala, Don Anushka Sandaruwan; Whang, Ilson; Lee, Sukkyoung; Kim, Hyowon; Zoysa, Mahanama De; Oh, Chulhong; Kang, Do-Hyung; Lee, Jehee

    2014-09-01

    Immune signaling cascades have an indispensable role in the host defense of almost all the organisms. Tumor necrosis factor (TNF) signaling is considered as a prominent signaling pathway in vertebrate as well as invertebrate species. Within the signaling cascade, TNF receptor-associated factor (TRAF) and TNF receptor-associated protein (TTRAP) has been shown to have a crucial role in the modulation of immune signaling in animals. Here, we attempted to characterize a novel molluskan ortholog of TTRAP (AbTTRAP) from disk abalone (Haliotis discus discus) and analyzed its expression levels under pathogenic stress. The complete coding sequence of AbTTRAP consisted of 1071 nucleotides, coding for a 357 amino acid peptide, with a predicted molecular mass of 40 kDa. According to our in-silico analysis, AbTTRAP resembled the typical TTRAP domain architecture, including a 5'-tyrosyl DNA phosphodiesterase domain. Moreover, phylogenetic analysis revealed its common ancestral invertebrate origin, where AbTTRAP was clustered with molluskan counterparts. Quantitative real time PCR showed universally distributed expression of AbTTRAP in selected tissues of abalone, from which more prominent expression was detected in hemocytes. Upon stimulation with two pathogen-derived mitogens, lipopolysaccharide (LPS) and polyinosinic:polycytidylic acid (poly I:C), transcript levels of AbTTRAP in hemocytes and gill tissues were differentially modulated with time. In addition, the recombinant protein of AbTTRAP exhibited prominent endonuclease activity against abalone genomic DNA, which was enhanced by the presence of Mg(2+) in the medium. Collectively, these results reinforce the existence of the TNF signaling cascade in mollusks like disk abalone, further implicating the putative regulatory behavior of TTRAP in invertebrate host pathology. Copyright © 2014 Elsevier Ltd. All rights reserved.

  1. Effect of air-temperature and diet composition on the drying process of pellets for japanese abalone (Haliotis discus hannai) feeding

    OpenAIRE

    Vega-Gálvez, Antonio; Lemus-Mondaca, Roberto; Plaza-Sáez, Daniel; Pérez-Won, Mario

    2011-01-01

    The aim of this research was to study the effect of air-temperature and diet composition on the mass transfer kinetics during the drying process of pellets used for Japanese Abalone (Haliotis discus hannai) feeding. In the experimental design, three temperatures were used for convective drying, as well as three different diet compositions (Diets A, B and C), in which the amount of fishmeal, spirulin, algae, fish oil and cornstarch varied. The water diffusion coefficient of the pellets was det...

  2. Sperm Donation and the Right to Privacy.

    Science.gov (United States)

    Hallich, Oliver

    2017-07-01

    Sperm donation is an increasingly common method of assisted reproduction. In the debate on sperm donation, the right to privacy - construed as a right that refers to the limits of the realm of information to which others have access - plays a pivotal role with regard to two questions. The first question is whether the sperm donor's right to privacy implies his right to retain his anonymity, the second is whether the gamete recipients' right to privacy entitles them to withhold information about the circumstances of their conception from their donor-conceived offspring. In this contribution, I tackle these two interrelated questions. In part (1), I defend the view that there is a prima facie right of sperm donors to remain anonymous. Part (2) widens the perspective by taking into consideration the welfare of donor-conceived offspring. I argue that anonymity may harm the child only if the gametes' recipients decide to disclose information about the circumstances of her birth to the child. Non-disclosure of these circumstances, however, is morally problematic because it may not necessarily harm, but wrong the child. In section (3), I attempt to rebut some arguments in defense of non-disclosure. In part (4), I defend the view that the best practice of sperm donation would be 'direct donation', i.e. that the identity of the donor is known from the time of conception. Part (5) concludes.

  3. Sperm length evolution in the fungus-growing ants

    DEFF Research Database (Denmark)

    Baer, B.; Dijkstra, M. B.; Mueller, U. G.

    2009-01-01

    Eusocial insects offer special opportunities for the comparative study of sperm traits because sperm competition is absent (in species with obligatory monandry) or constrained (in lineages where queens mate multiply but never remate later in life). We measured sperm length in 19 species of fungus......-growing ants, representing 9 of the 12 recognized genera, and mapped these onto the ant phylogeny. We show that average sperm length across species is highly variable and decreases with mature colony size in basal genera with singly mated queens, suggesting that sperm production or storage constraints affect...... the evolution of sperm length. Sperm length does not decrease further in multiply mating leaf-cutting ants, despite substantial further increases in colony size. In a combined analysis, sexual dimorphism explained 63.1% of the variance in sperm length between species. As colony size was not a significant...

  4. Short communication Sperm DNA damage in relation to lipid ...

    African Journals Online (AJOL)

    Leyland Fraser

    2017-03-08

    LPO) and sperm DNA damage following freezing-thawing of boar semen in different extenders. The comet assay was used to measure the extent of sperm DNA damage in a cryoprotectant-free extender or in cryoprotectant-based ...

  5. Sperm viability staining in ecology and evolution: potential pitfalls

    DEFF Research Database (Denmark)

    Holman, Luke

    2009-01-01

    The causes and consequences of variation in sperm quality, survival and ageing are active areas of research in ecology and evolution. In order to address these topics, many recent studies have measured sperm viability using fluorescent staining. Although sperm viability staining has produced...... a number of interesting results, it has some potential pitfalls that have rarely been discussed. In the present paper, I review the major findings of ecology and evolution studies employing sperm viability staining and outline the method's principle limitations. The key problem is that the viability assay...... may itself kill sperm, which is likely to confound many common experimental designs in addition to producing artificially low estimates of sperm viability. I further suggest that sperm number should be routinely measured in sperm viability studies, as it may be an important but overlooked source...

  6. Prudent sperm use by leaf-cutter ant queens

    DEFF Research Database (Denmark)

    Den Boer, Susanne Petronella A; Baer, Boris; Dreier, Stéphanie Agnès Jeanine

    2009-01-01

    In many species, females store sperm between copulation and egg fertilization, but the consequences of sperm storage and patterns of sperm use for female life history and reproductive success have not been investigated in great detail. In hymenopteran insect societies (ants, bees, wasps......), reproduction is usually monopolized by one or relatively few queens, who mate only during a brief period early in life and store sperm for later use. The queens of some ants are particularly long-lived and have the potential to produce millions of offspring during their life. To do so, queens store many sperm...... cells, and this sperm must remain viable throughout the years of storage. Queens should also be under strong selection to use stored sperm prudently when fertilizing eggs. We used the leaf-cutter ant Atta colombica to investigate the dynamics of sperm use during egg fertilization. We show that queens...

  7. Pulmonary exposure to carbonaceous nanomaterials and sperm quality

    DEFF Research Database (Denmark)

    Skovmand, Astrid; Lauvas, Anna Jacobsen; Christensen, Preben

    2018-01-01

    inflammation is a potential modulator of endocrine function. The aim of this study was to investigate the effects of pulmonary exposure to carbonaceous nanomaterials on sperm quality parameters in an experimental mouse model.Methods: Effects on sperm quality after pulmonary inflammation induced by carbonaceous....... Pulmonary inflammation was determined by differential cell count in bronchoalveolar lavage fluid. Epididymal sperm concentration and motility were measured by computer-assisted sperm analysis. Epididymal sperm viability and morphological abnormalities were assessed manually using Hoechst 33,342/PI...... flourescent and Spermac staining, respectively. Epididymal sperm were assessed with regard to sperm DNA integrity (damage). Daily sperm production was measured in the testis, and testosterone levels were measured in blood plasma by ELISA.Results: Neutrophil numbers in the bronchoalveolar fluid showed...

  8. Gulf of Mexico sperm whale photo-ID catalog

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Photo-identification data on sperm whales occupying the north central Gulf of Mexico have been collected during vessel surveys. Photographs of sperm whales are taken...

  9. Intracytoplasmic sperm injection (ICSI) and chromosomally abnormal spermatozoa

    NARCIS (Netherlands)

    P.A. in 't Veld; F.J.M. Broekmans (Frank); H.F. de France; P.L. Pearson; M.H. Pieters; R.J. van Kooij

    1997-01-01

    textabstractAn infertile couple was referred for intracytoplasmic sperm injection (ICSI) because of primary infertility and oligoasthenoteratozoospermia (OAT) in the male. It was observed that although the sperm cells presented with an unusual head size and multiple

  10. The feasibility of bomb radiocarbon analysis to support an age-at-length relationship for red abalone, Haliotis rufescens Swainson in northern California

    Energy Technology Data Exchange (ETDEWEB)

    Leaf, R T; Andrews, A H; Cailliet, G M; Brown, T A

    2009-01-07

    Analysis of bomb generated radiocarbon ({sup 14}C) changes in a red abalone, Haliotis rufescens Swainson shell was used to investigate age-at-length relationships derived from data from a previous multi-year, multi-site tag-recapture study. Shell carbonate was extracted from four successive growth trajectory locations in a single shell with a length of 251 mm MSL. Extraction locations were based on VBGF predictions and chosen to span the initial rise of the {sup 14}C-bomb pulse that is known to have occurred in surface ocean waters during 1958 {+-} 1 y in the northeast Pacific. The close temporal correspondence of the red abalone sample series to regional {Delta}{sup 14}C records demonstrated the utility of the technique for validating age-at-length relationships for the red abalone. The findings provided support for a mean VBGF derived age of 32 y (range 30 to 33 y) for the specimen; however, the analysis of {sup 14}C data indicated that the specimen could be older.

  11. Molecular insights of the first gastropod TLR counterpart from disk abalone (Haliotis discus discus), revealing its transcriptional modulation under pathogenic stress.

    Science.gov (United States)

    Elvitigala, Don Anushka Sandaruwan; Premachandra, H K A; Whang, Ilson; Nam, Bo-Hye; Lee, Jehee

    2013-08-01

    Toll-like receptors (TLRs) are well-characterized pattern recognition receptors of innate immunity, known to induce immune responses against the pathogens by interacting with evolutionarily conserved pathogen-associated molecular patterns (PAMPs). In this study, a novel TLR homolog from disk abalone (Haliotis discus discus) was identified and characterized at molecular level. The open reading frame (ORF) of AbTLR is 3804 bp in length and encodes a 1268 amino acid peptide with a calculated molecular mass of 143.5 kDa. The deduced protein shows typical TLR domain architecture, with leucine rich repeats (LRR) and the toll-interleukin receptor (TIR) domain. Phylogenetic analysis revealed a close evolutionary relationship for AbTLR to its invertebrate counterparts, with close clustering to the molluscan homologs. Quantitative real-time PCR detected ubiquitous transcription of AbTLR in healthy tissues, but with highest levels in hemocytes. Differential transcriptional modulation of AbTLR was observed in abalone hemocytes and gills upon immune challenge, whereby Vibrio parahaemolyticus and purified lipopolysaccharide (LPS) enhanced the transcript level prominently. In addition, the viral hemorrhagic septicemia virus induced AbTLR transcription in hemocytes and gills, representing the first evidence of viral-induced immune response in mollusks to date. Collectively, our findings support a putative role for AbTLR in abalone antiviral and antibacterial defense. Copyright © 2013 Elsevier Ltd. All rights reserved.

  12. Transcriptome-Wide Single Nucleotide Polymorphisms (SNPs for Abalone (Haliotis midae: Validation and Application Using GoldenGate Medium-Throughput Genotyping Assays

    Directory of Open Access Journals (Sweden)

    Rouvay Roodt-Wilding

    2013-09-01

    Full Text Available Haliotis midae is one of the most valuable commercial abalone species in the world, but is highly vulnerable, due to exploitation, habitat destruction and predation. In order to preserve wild and cultured stocks, genetic management and improvement of the species has become crucial. Fundamental to this is the availability and employment of molecular markers, such as microsatellites and Single Nucleotide Polymorphisms (SNPs . Transcriptome sequences generated through sequencing-by-synthesis technology were utilized for the in vitro and in silico identification of 505 putative SNPs from a total of 316 selected contigs. A subset of 234 SNPs were further validated and characterized in wild and cultured abalone using two Illumina GoldenGate genotyping assays. Combined with VeraCode technology, this genotyping platform yielded a 65%−69% conversion rate (percentage polymorphic markers with a global genotyping success rate of 76%−85% and provided a viable means for validating SNP markers in a non-model species. The utility of 31 of the validated SNPs in population structure analysis was confirmed, while a large number of SNPs (174 were shown to be informative and are, thus, good candidates for linkage map construction. The non-synonymous SNPs (50 located in coding regions of genes that showed similarities with known proteins will also be useful for genetic applications, such as the marker-assisted selection of genes of relevance to abalone aquaculture.

  13. The effect of different polychlorinated biphenyls on two aquatic models, the green alga Pseudokirchneriella subcapitata and the haemocytes from the European abalone Haliotis tuberculata.

    Science.gov (United States)

    Halm-Lemeille, Marie-Pierre; Abbaszadeh Fard, Elham; Latire, Thomas; Ferard, Jean-François; Costil, Katherine; Lebel, Jean-Marc; Bureau, Ronan; Serpentini, Antoine

    2014-09-01

    The present study was conducted to determine the toxicity of different polychlorinated biphenyls (PCBs) on the green algae, Pseudokirchneriella subcapitata and the haemocytes from the European abalone, Haliotis tuberculata. Using the algal growth inhibition test, the green algae median Effective Concentration (EC50) values ranged from 0.34μM for PCB28 to more than 100μM for PCBs 101 and 153. Considering the MTT viability test, the abalone EC50 values ranged from 1.67μM for PCB153 to 89μM for PCB28. Our results in contrast to previous observation in vertebrates did not show significant differences between the dioxin like- and non dioxin like-PCBs toxicities regardless of the model used. However, our results demonstrated that the toxicities of PCBs were species dependent. For example, PCB28 was the most toxic compound for P. subcapitata whereas PCBs 1, 180 and 153 were less toxic for that species. On the contrary, PCB153 was reported as the most toxic for H. tuberculata haemocytes and PCB28 the least toxic. To investigate the mode of action of these compounds, we used an in silico method. Our results suggested that PCBs have a non-specific mode of action (e.g., narcosis) on green algae, and another mode of action, probably more specific than narcosis, was reported for PCBs on the abalone haemocytes. Copyright © 2014 Elsevier Ltd. All rights reserved.

  14. Does breastfeeding influence future sperm quality and reproductive hormones?

    DEFF Research Database (Denmark)

    Laustsen, J M; Jensen, M S; Thulstrup, Ane Marie

    2011-01-01

    was not statistically significantly associated with sperm concentration, total sperm count, sperm motility or morphology, oligozoospermia, follicle-stimulating hormone, inhibin B, luteinizing hormone, sex hormone-binding globulin (SHBG), the calculated level of free testosterone, free oestradiol, the free testosterone...... testosterone nor free oestradiol was different between the two groups. This study shows no association between breastfeeding and sperm quality or reproductive hormones and a strong association is unlikely. A larger study would be needed to detect more subtle effects....

  15. Study of sperm proteins in different mammalian species

    OpenAIRE

    Pohlová, Alžběta

    2016-01-01

    Reproduction is an essential feature of all animals and a fundamental step to produce new generations. Study of sperm proteins is crucial for understanding of the sperm-egg recognition. We searched out sperm surface proteins involving in the zona pellucida (ZP) binding and studied whether these proteins are preserved throughout mammalian species. Indirect immunofluorescent technique was used to test a panel of monoclonal antibodies prepared against boar sperm surface proteins on spermatozoa o...

  16. Sperm retrieval outcomes with microdissection testicular sperm extraction (micro-TESE) in men with cryptozoospermia.

    Science.gov (United States)

    Alrabeeah, K; Wachter, A; Phillips, S; Cohen, B; Al-Hathal, N; Zini, A

    2015-05-01

    Several studies support of the use of testicular rather than ejaculated spermatozoa for intracytoplasmic sperm injection (ICSI) in couples with virtual azoospermia or cryptozoospermia, although this approach remains controversial. We sought to evaluate sperm retrieval outcomes with microdissection testicular sperm extraction (micro-TESE) in men with cryptozoospermia. We conducted a retrospective study of 24 consecutive micro-TESEs in men with cryptozoospermia. We also evaluated the outcomes of seven consecutive TESAs (testicular sperm aspiration) in cryptozoospermic men during the same time period (January 2007 and September 2014). Micro-TESE and TESA were performed on the day prior to ICSI. Final assessment of sperm recovery (reported on the day of ICSI) was recorded as (i) successful (available spermatozoa for ICSI) or (ii) unsuccessful (no spermatozoa for ICSI). The decision to perform a unilateral or bilateral micro-TESE was guided by the intra-operative evaluation of sperm recovery from the first testicle. A unilateral procedure was performed in 87.5% (21/24) and 57% (4/7) of the micro-TESE and TESA cohorts, respectively. Sperm recovery was successful in 96% (23/24) of the men who underwent micro-TESE and 43% (3/7) of the men who underwent TESA (p TESE and TESA groups were comparable [33% (6/18) and 50% (1/2), respectively]. The data indicate that micro-TESE is a highly successful sperm retrieval technique for men with cryptozoospermia and few of these men will require a bilateral procedure. Moreover, sperm retrieval rates are higher with micro-TESE than TESA in this group of men. © 2015 American Society of Andrology and European Academy of Andrology.

  17. Sperm cell surface dynamics during activation and fertilization

    NARCIS (Netherlands)

    Boerke, A.|info:eu-repo/dai/nl/304822922

    2013-01-01

    Before the sperm cell can reach the oocyte it needs to be activated and to undergo a series of preparative steps. The sperm surface dynamics was studied in relation to this activation process and the modifications and removal of sperm surface components havebeen investigated. Bicarbonate-induced

  18. Evolutionary genetics and ecology of sperm-dependent parthenogenesis

    NARCIS (Netherlands)

    Beukeboom, L.W.; Vrijenhoek, R.C.

    1998-01-01

    Sperm-dependent (or pseudogamous) forms of parthenogenetic reproduction occur in a wide variety of animals. Inheritance is typically clonal and matroclinous (of female descent), but sperm are needed to initiate normal development. As opposed to true parthenogenesis (i.e., sperm-independent

  19. Effect of 4-nonylphenol on the sperm dynamic parameters ...

    African Journals Online (AJOL)

    4-Nonylphenol (NP) is a compound that causes endocrine disruption and affects sperm quality of mammals and fish. However, the effects of NP on the sperm and fertilization rate of amphibians remain unknown. This study investigates the in vivo and in vitro effects of NP on the sperm dynamic parameters and fertilization ...

  20. Sperm storage and duration of fertility in female ostriches ( Struthio ...

    African Journals Online (AJOL)

    , the duration of sperm storage and the fertile period following separation of sexes were investigated by egg break-out and by counting the sperm in the perivitelline membrane (spermOPVL) above the germinal disc (GD) region. Fertilisation ...

  1. LOCALIZATION OF SP22 ON HUMAN SPERM OF DIFFERING QUALITY

    Science.gov (United States)

    LOCALIZATION OF SP22 ON HUMAN SPERM OF DIFFERING QUALITY. AE Lavers*1, GR Klinefelter2, DW Hamilton1, KP Roberts1, 1University of Minnesota, Minneapolis, MN and 2US EPA, Research Triangle Park, NC. SP22 is a sperm membrane protein that has been implicated in sperm function d...

  2. Sperm DNA fragmentation, recurrent implantation failure and recurrent miscarriage

    Directory of Open Access Journals (Sweden)

    Carol Coughlan

    2015-01-01

    Full Text Available Evidence is increasing that the integrity of sperm DNA may also be related to implantation failure and recurrent miscarriage (RM. To investigate this, the sperm DNA fragmentation in partners of 35 women with recurrent implantation failure (RIF following in vitro fertilization, 16 women diagnosed with RM and seven recent fathers (control were examined. Sperm were examined pre- and post-density centrifugation by the sperm chromatin dispersion (SCD test and the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL assay. There were no significant differences in the age of either partner or sperm concentration, motility or morphology between three groups. Moreover, there were no obvious differences in sperm DNA fragmentation measured by either test. However, whilst on average sperm DNA fragmentation in all groups was statistically lower in prepared sperm when measured by the SCD test, this was not seen with the results from the TUNEL assay. These results do not support the hypothesis that sperm DNA fragmentation is an important cause of RIF or RM, or that sperm DNA integrity testing has value in such patients. It also highlights significant differences between test methodologies and sperm preparation methods in interpreting the data from sperm DNA fragmentation tests.

  3. Ovarian fluid of receptive females enhances sperm velocity

    Science.gov (United States)

    Gasparini, Clelia; Andreatta, Gabriele; Pilastro, Andrea

    2012-05-01

    The females of several internal fertilizers are able to store sperm for a long time, reducing the risk of sperm limitation. However, it also means that males can attempt to mate outside females' receptive period, potentially increasing the level of sperm competition and exacerbating sexual conflict over mating. The guppy ( Poecilia reticulata), an internally fertilizing fish, is a model system of such competition and conflict. Female guppies accept courtship and mate consensually only during receptive periods of the ovarian cycle but receive approximately one (mostly forced) mating attempt per minute both during and outside their sexually receptive phase. In addition, females can store viable sperm for months. We expected that guppy females would disfavour sperm received during their unreceptive period, possibly by modulating the quality and/or quantity of the components present in the ovarian fluid (OF) over the breeding cycle. Ovarian fluid has been shown to affect sperm velocity, a determinant of sperm competition success in this and other fishes. We found that in vitro sperm velocity is slower in OF collected from unreceptive females than in OF from receptive females. Visual stimulation with a potential partner prior to collection did not significantly affect in vitro sperm velocity. These results suggest that sperm received by unreceptive females may be disfavoured as sperm velocity likely affects the migration process and the number of sperm that reach storage sites.

  4. Thinking though eggs, sperm and embryos- diffractive readings at the fertility clinic and sperm bank

    DEFF Research Database (Denmark)

    Willum Adrian, Stine

    questions may an analytical starting point in eggs, sperm and embryos, enable me to ask, and what diffractive stories of creation can I tell? These questions not only intrigue me, because I am deeply fascinated about new creation stories that may be told when assisted reproductive technologies are at use...... stories are told about egg, sperm and embryos (Adrian 2006), my current work has moved to look at negotiations of ethical borders and boundaries in relation to globalization of Danish donor sperm. This move in research questions is not only empirically based. It is also a shift indebted to my...

  5. Posttesticular sperm maturation, infertility, and hypercholesterolemia

    Directory of Open Access Journals (Sweden)

    Marjorie Whitfield

    2015-01-01

    Full Text Available Cholesterol is a key molecule in the mammalian physiology of especial particular importance for the reproductive system as it is the common precursor for steroid hormone synthesis. Cholesterol is also a recognized modulator of sperm functions, not only at the level of gametogenesis. Cholesterol homeostasis regulation is crucial for posttesticular sperm maturation, and imbalanced cholesterol levels may particularly affect these posttesticular events. Metabolic lipid disorders (dyslipidemia affect male fertility but are most of the time studied from the angle of endocrine/testicular consequences. This review will focus on the deleterious effects of a particular dyslipidemia, i.e., hypercholesterolemia, on posttesticular maturation of mammalian spermatozoa.

  6. Involvement of opsins in mammalian sperm thermotaxis

    Science.gov (United States)

    Pérez-Cerezales, Serafín; Boryshpolets, Sergii; Afanzar, Oshri; Brandis, Alexander; Nevo, Reinat; Kiss, Vladimir; Eisenbach, Michael

    2015-01-01

    A unique characteristic of mammalian sperm thermotaxis is extreme temperature sensitivity, manifested by the capacity of spermatozoa to respond to temperature changes of functional approaches that opsins are present in human and mouse spermatozoa at specific sites, which depend on the species and the opsin type, and that they are involved in sperm thermotaxis via two signalling pathways—the phospholipase C and the cyclic-nucleotide pathways. Our results suggest that, depending on the context and the tissue, mammalian opsins act not only as photosensors but also as thermosensors. PMID:26537127

  7. Fate of the sperm mitochondria, and the incorporation, conversion, and disassembly of the sperm tail structures during bovine fertilization.

    Science.gov (United States)

    Sutovsky, P; Navara, C S; Schatten, G

    1996-12-01

    Sperm incorporation and the conversion of the sperm-derived components into zygotic structures during in vitro fertilization of bovine oocytes was explored by combining ultrastructural studies with observations of the fertilizing sperm tagged with a mitochondrion-specific vital dye MitoTracker green FM. The zygotes fertilized by the MitoTracker-labeled sperm were fixed at various times after fertilization and then processed for immunocytochemistry to examine the distribution of DNA, microtubules, and sperm tail components, including the fibrous sheath and axonemal microtubules. We show here that the complete incorporation of the sperm, but not sperm-oocyte binding and oocyte activation, depends upon the integrity of oocyte microfilaments and is inhibited by the microfilament disrupter cytochalasin B. After sperm incorporation, the mitochondria are displaced from the sperm's connecting piece, and the sperm centriole is exposed to the egg cytoplasm. This event is followed by the formation of the microtubule-based sperm aster, which is responsible for the union of male and female pronuclei. Concomitantly, the major structure of the sperm principal piece, the fibrous sheath, disappears. After the first mitosis, the compact mitochondrial sheath can be seen in one of the blastomeres. An aggregate of the sperm mitochondria is observed at the entry of the second mitosis, although they remain in the vicinity of the nucleus and can later be seen at one pole of the metaphase spindle. The mitochondrial cluster is occasionally found in one of the blastomeres in the early-stage four-cell embryos, but it is no longer detected by the beginning of the third mitotic cycle. These data suggest that the disassembly of the sperm tail during bovine fertilization occurs as a series of precisely orchestrated events involving the destruction (fibrous sheath and mitochondrial sheath) and transformation (DNA, sperm centriole) of particular sperm structures into zygotic and embryonic

  8. Structural aspects of the solvation shell of lysine and acetylated lysine: A Car-Parrinello and classical molecular dynamics investigation

    International Nuclear Information System (INIS)

    Carnevale, V.; Raugei, S.

    2009-01-01

    Lysine acetylation is a post-translational modification, which modulates the affinity of protein-protein and/or protein-DNA complexes. Its crucial role as a switch in signaling pathways highlights the relevance of charged chemical groups in determining the interactions between water and biomolecules. A great effort has been recently devoted to assess the reliability of classical molecular dynamics simulations in describing the solvation properties of charged moieties. In the spirit of these investigations, we performed classical and Car-Parrinello molecular dynamics simulations on lysine and acetylated-lysine in aqueous solution. A comparative analysis between the two computational schemes is presented with a focus on the first solvation shell of the charged groups. An accurate structural analysis unveils subtle, yet statistically significant, differences which are discussed in connection to the significant electronic density charge transfer occurring between the solute and the surrounding water molecules.

  9. Effect of transfection and co-incubation of bovine sperm with exogenous DNA on sperm quality and functional parameters for its use in sperm-mediated gene transfer.

    Science.gov (United States)

    Arias, María Elena; Sánchez-Villalba, Esther; Delgado, Andrea; Felmer, Ricardo

    2017-02-01

    Sperm-mediated gene transfer (SMGT) is based on the capacity of sperm to bind exogenous DNA and transfer it into the oocyte during fertilization. In bovines, the progress of this technology has been slow due to the poor reproducibility and efficiency of the production of transgenic embryos. The aim of the present study was to evaluate the effects of different sperm transfection systems on the quality and functional parameters of sperm. Additionally, the ability of sperm to bind and incorporate exogenous DNA was assessed. These analyses were carried out by flow cytometry and confocal fluorescence microscopy, and motility parameters were also evaluated by computer-assisted sperm analysis (CASA). Transfection was carried out using complexes of plasmid DNA with Lipofectamine, SuperFect and TurboFect for 0.5, 1, 2 or 4 h. The results showed that all of the transfection treatments promoted sperm binding and incorporation of exogenous DNA, similar to sperm incorporation of DNA alone, without affecting the viability. Nevertheless, the treatments and incubation times significantly affected the motility parameters, although no effect on the integrity of DNA or the levels of reactive oxygen species (ROS) was observed. Additionally, we observed that transfection using SuperFect and TurboFect negatively affected the acrosome integrity, and TurboFect affected the mitochondrial membrane potential of sperm. In conclusion, we demonstrated binding and incorporation of exogenous DNA by sperm after transfection and confirmed the capacity of sperm to spontaneously incorporate exogenous DNA. These findings will allow the establishment of the most appropriate method [intracytoplasmic sperm injection (ICSI) or in vitro fertilization (IVF)] of generating transgenic embryos via SMGT based on the fertilization capacity of transfected sperm.

  10. Effects of waterborne Cu and Cd on anti-oxidative response, lipid peroxidation and heavy metals accumulation in abalone Haliotis discus hannai ino

    Science.gov (United States)

    Lei, Yanju; Zhang, Wenbing; Xu, Wei; Zhang, Yanjiao; Zhou, Huihui; Mai, Kangsen

    2015-06-01

    The aim of this study was to compare the effects of waterborne copper (Cu) and cadmium (Cd) on survival, anti-oxidative response, lipid peroxidation and metal accumulation in abalone Haliotis discus hannai. Experimental animals (initial weight: 7.49 g ± 0.01 g) were exposed to graded concentrations of waterborne Cu (0.02, 0.04, 0.06, 0.08 mg L-1) or Cd (0.025, 0.05, 0.25, 0.5 mg L-1) for 28 days, respectively. Activities of the anti-oxidative enzymes (catalase, CAT; superoxide dismutase, SOD; glutathione peroxidases, GPx; glutathione S-transferase, GST), contents of the reduced glutathione (GSH) and malondiadehyde (MDA) in the hepatopancreas, and metal accumulation in hepatopancreas and muscles were analyzed after 0, 1, 3, 6, 10, 15, 21, 28 days of metal exposure, respectively. Results showed that 0.04 mg L-1, 0.06 mg L--1 and 0.08 mg L-1 Cu caused 100% death of abalone on the 21st, 10th and 6th day, respectively. However, no dead abalone was found during the 28-day waterborne Cd exposure at all experimental concentrations. Generally, activities of SOD and GST in hepatopancreas under all Cu concentrations followed a decrease trend as the exposure time prolonged. However, these activities were firstly increased and then decreased to the control level and increased again during Cd exposure. Activities of CAT in all Cu exposure treatments were higher than those in the control. These activities were firstly increased and then decreased to the control level and increased again during Cd exposure. Contents of MDA in hepatopancreas in all Cu treatments significantly increased first and then decreased to the control level. However, the MDA contents in hepatopancreas were not significantly changed during the 28-day Cd exposure. The metals accumulation in both hepatopancreas and muscles of abalone significantly increased with the increase of waterborne metals concentration and exposure time. These results indicated that H. discus hannai has a positive anti-oxidative defense

  11. Two duplicated chicken-type lysozyme genes in disc abalone Haliotis discus discus: molecular aspects in relevance to structure, genomic organization, mRNA expression and bacteriolytic function.

    Science.gov (United States)

    Umasuthan, Navaneethaiyer; Bathige, S D N K; Kasthuri, Saranya Revathy; Wan, Qiang; Whang, Ilson; Lee, Jehee

    2013-08-01

    Lysozymes are crucial antibacterial proteins that are associated with catalytic cleavage of peptidoglycan and subsequent bacteriolysis. The present study describes the identification of two lysozyme genes from disc abalone Haliotis discus discus and their characterization at sequence-, genomic-, transcriptional- and functional-levels. Two cDNAs and BAC clones bearing lysozyme genes were isolated from abalone transcriptome and BAC genomic libraries, respectively and sequences were determined. Corresponding deduced amino acid sequences harbored a chicken-type lysozyme (LysC) family profile and exhibited conserved characteristics of LysC family members including active residues (Glu and Asp) and GS(S/T)DYGIFQINS motif suggested that they are LysC counterparts in disc abalone and designated as abLysC1 and abLysC2. While abLysC1 represented the homolog recently reported in Ezo abalone [1], abLysC2 shared significant identity with LysC homologs. Unlike other vertebrate LysCs, coding sequence of abLysCs were distributed within five exons interrupted by four introns. Both abLysCs revealed a broader mRNA distribution with highest levels in mantle (abLysC1) and hepatopancreas (abLysC2) suggesting their likely main role in defense and digestion, respectively. Investigation of temporal transcriptional profiles post-LPS and -pathogen challenges revealed induced-responses of abLysCs in gills and hemocytes. The in vitro muramidase activity of purified recombinant (r) abLysCs proteins was evaluated, and findings indicated that they are active in acidic pH range (3.5-6.5) and over a broad temperature range (20-60 °C) and influenced by ionic strength. When the antibacterial spectra of (r)abLysCs were examined, they displayed differential activities against both Gram positive and Gram negative strains providing evidence for their involvement in bacteriolytic function in abalone physiology. Copyright © 2013 Elsevier Ltd. All rights reserved.

  12. The maintenance of sperm variability: context-dependent selection on sperm morphology in a broadcast spawning invertebrate.

    Science.gov (United States)

    Johnson, Darren W; Monro, Keyne; Marshall, Dustin J

    2013-05-01

    Why are sperm so variable despite having a singular, critical function and an intimate relationship with fitness? A key to understanding the evolution of sperm morphology is identifying which traits enable sperm to be successful fertilizers. Several sperm traits (e.g., tail length, overall size) are implicated in sperm performance, but the benefits of these traits are likely to be highly context dependent. Here, we examined phenotypic selection on sperm morphology of a broadcast spawning tube worm (Galeolaria gemineoa). We conducted laboratory experiments to measure the relationship between average sperm morphology and relative fertilization success across a range of sperm environments that were designed to approximate the range of sperm concentrations and ages encountered by eggs in nature. We found that the strength and form of multivariate selection varied substantially across our environmental gradients. Sperm with long tails and small heads were favored in high-concentration environments, whereas sperm with long heads were favored at low concentrations and old ages. We suggest variation in the local fertilization environment and resulting differences in selection can preserve variability in sperm morphology both within and among males. © 2012 The Author(s). Evolution © 2012 The Society for the Study of Evolution.

  13. Exogenous neurotensin modulates sperm function in Japanese Black cattle.

    Science.gov (United States)

    Umezu, Kohei; Hiradate, Yuuki; Oikawa, Toshinori; Ishiguro, Hirotoshi; Numabe, Takashi; Hara, Kenshiro; Tanemura, Kentaro

    2016-08-25

    Recently, the conception rates after artificial insemination have been pointed out to decline continuously. To overcome this problem, the control of frozen and thawed sperm quality is required. However, the mechanism of bovine sperm functional regulation is still largely unknown. In mammals, the ejaculated sperm are capable of showing fertilizing ability during migration in the female reproductive organs. It is well known that these female organs secrete several factors contributing to sperm capacitation. We previously reported that neurotensin (NT) secreted from the oviduct and cumulus cells enhanced sperm capacitation and acrosome reaction in mice. In this study, we confirmed the expression of the NT receptor (NTR1) in the bovine sperm neck region and the secretion of NT in the bovine uterus and oviduct. The similar expression patterns of NT and NTR1 suggests a conserved mechanism of sperm functional regulation between mouse and cattle. Thus, we examined the effects of exogenous NT on the bovine sperm functions. First, we showed that NT induced sperm protein tyrosine phosphorylation in a dose-dependent manner, suggesting that NT enhances sperm capacitation. Second, we showed that NT induced acrosome reactions of capacitated sperm in a dose-dependent manner, suggesting that NT facilitates acrosome reaction. Finally, we used a computer-aided sperm analysis system to show that NT did not have a great effect on sperm motility. These results suggest that NT acts as a facilitator of sperm capacitation and acrosome reaction in the female reproductive tracts in cattle, highlighting the importance of NT-mediated signaling to regulate sperm functions.

  14. Improving sperm banking efficiency in endangered species through the use of a sperm selection method in brown bear (Ursus arctos) thawed sperm.

    Science.gov (United States)

    Anel-Lopez, L; Ortega-Ferrusola, C; Álvarez, M; Borragán, S; Chamorro, C; Peña, F J; Morrell, J; Anel, L; de Paz, P

    2017-06-26

    Sperm selection methods such as Single Layer Centrifugation (SLC) have been demonstrated to be a useful tool to improve the quality of sperm samples and therefore to increase the efficiency of other artificial reproductive techniques in several species. This procedure could help to improve the quality of genetic resource banks, which is essential for endangered species. In contrast, these sperm selection methods are optimized and focused on farm animals, where the recovery task is not as important as in endangered species because of their higher sperm availability. The aim of this study was to evaluate two centrifugation methods (300 x g/20 min and 600 x g/10 min) and three concentrations of SLC media (Androcoll-Bear -80, 65 and 50%) to optimise the procedure in order to recover as many sperm with the highest quality as possible. Sperm morphology could be important in the hydrodynamic relationship between the cell and centrifugation medium and thus the effect of sperm head morphometry on sperm yield and its hydrodynamic relationship were studied. The samples selected with Androcoll-Bear 65% showed a very good yield (53.1 ± 2.9) although the yield from Androcoll-Bear 80% was lower (19.3 ± 3.3). The latter showed higher values of motility than the control immediately after post-thawing selection. However, both concentrations of colloid (65 and 80%) showed higher values of viable sperm and viable sperm with intact acrosome than the control. After an incubation of 2 h at 37 °C, the samples from Androcoll-Bear 80% had higher kinematics and proportion of viable sperm with intact acrosome. In the morphometric analysis, the sperm selected by the Androcoll-Bear 80% showed a head with a bigger area which was more elongated than the sperm from other treatments. We conclude that sperm selection with Androcoll-Bear at either 65% or 80% is a suitable technique that allows a sperm population with better quality than the initial sample to be obtained. We recommend the

  15. Bacteriophage phi11 lysin: Physicochemical characterization and comparison with phage phi80α lysin.

    Science.gov (United States)

    Filatova, Lyubov Y; Donovan, David M; Foster-Frey, Juli; Pugachev, Vladimir G; Dmitrieva, Natalia F; Chubar, Tatiana A; Klyachko, Natalia L; Kabanov, Alexander V

    2015-06-01

    Phage lytic enzymes are promising antimicrobial agents. Lysins of phages phi11 (LysPhi11) and phi80α (LysPhi80α) can lyse (destroy) cells of antibiotic-resistant strains of Staphylococcus aureus. Stability of enzymes is one of the parameters making their practical use possible. The objectives of the study were to investigate the stability of lysins of phages phi11 and phi80α in storage and functioning conditions, to identify optimum storage conditions and causes of inactivation. Stability of the recombinant LysPhi11 and LysPhi80α was studied using turbidimetry. CD-spectroscopy, dynamic light scattering, and electrophoresis were used to identify causes of inactivation. At 37°C, pH 7.5 and concentration of NaCl not higher than 150mM, LysPhi11 molecules contain a high percentage of random coils (43%). However, in spite of this the enzyme has high activity (0.4-0.8OD600nms(-1)mg(-1)). In storage conditions (4°C and 22°C, pH 6.0-9.0, 10-500mM NaCl) LysPhi11 is inactivated by a monomolecular mechanism. The optimum storage conditions for LysPhi11 (4°C, pH 6.0-7.5, 10mM NaCl) were selected under which the time of the enzyme half-inactivation is 120-160 days. LysPhi80α stability is insufficient: at 37°C the enzyme loses half of its activity almost immediately; at 4°C and 22°C the time of half-inactivation of LysPhi80α varies in the range from several hours to 3 days. Despite the common properties in the manifestation of antistaphylococcal activity the kinetic behavior of the enzymes is different. LysPhi11 is a more promising candidate to be used as an antimicrobial agent. Copyright © 2015 Elsevier Inc. All rights reserved.

  16. Effect of sorafenib on sperm count and sperm motility in male Swiss albino mice

    Directory of Open Access Journals (Sweden)

    Surekha Devadasa Shetty

    2015-01-01

    Full Text Available The issue of male germ line mutagenesis and the effects on developmental defects in the next generation has become increasingly high profile over recent years. Mutagenic substance affects germinal cells in the testis. Since the cells are undergoing different phases of cell division and maturation, it is an ideal system to study the effect of chemotherapeutic agents. There are lacunae in the literature on the effect of sorafenib on gonadal function. With background, a study was planned to evaluate the effects of sorafenib on sperm count and sperm motility in male Swiss albino mice. Male Swiss albino mice were used for the study. The animals were segregated into control, positive control (PC and three treatment groups. PC received oral imatinib (100 mg/kg body weight and treatment groups received 25, 50, and 100 mg/kg body weight of sorafenib orally for 7 consecutive days at intervals of 24 h between two administrations. The control group remained in the home cage for an equal duration of time to match their corresponding treatment groups. The animals were sacrificed at the end of 1 st , 2 nd , 4 th , 5 th , 7 th , and 10 th weeks after the last exposure to drug, respectively. Sperm suspensions were prepared and introduced into a counting chamber. Total sperm count and motility were recorded. There was a significant decrease in sperm count and sperm motility by sorafenib which was comparable with the effect of PC imatinib. Sorafenib adversely affects sperm count and sperm motility which are reversible after discontinuation of treatment.

  17. Study of Sperm Parameters and Sperm Fertility in Mice were Exposed to Tamoxifen during Embryonic Development

    Directory of Open Access Journals (Sweden)

    J Soleimanirad

    2017-05-01

    Full Text Available Introduction: Tamoxifen is steroidal drug, which mainly treats breast cancer and also used to stimulate ovulation. The purpose of the present study was the evaluation of sperm parameters and fertility of mice whose mothers had received tamoxifen during pregnancy. Methods: In this study, 30 female and 15 male mice of NMRI were selected for mating. After mating female mice were randomly divided into two groups, the first group (control and second group (experimental. All of which contained 15 mice. From the day 13th day of pregnancy, experimental group has received tamoxifen with the dosage of 5 mg/kg for 7 days. After childbirth of the mated mice, male infants were selected. After reaching the age of puberty (6-8Weeks, adult mice were sacrificed by the cervical dislocation. After take sperm, sperm parameters (count, normality and motility, and sperm fertility was performed. In this study SPSS software and statistical t-test was used (p <0.001. Results: Studies showed that sperm parameters and sperm fertilization were significantly different. The number of sperm in the control group was 83.50±28.20 million, and in the experimental group was 60±14.14 million. There was a decrease in average sperm count in the experimental group compared with the control group (p <0.001. Our findings from in vitro fertilization culture media showed that embryos formation and oocyte disruption between control and experimental groups significantly different (p <0.001. Conclusion: The results showed that tamoxifen exposure during development can cause histological changes in the seminiferous tubules, which can lead to infertility.

  18. Antioxidant activity of maillard reaction products from lysine-glucose ...

    African Journals Online (AJOL)

    Maillard reaction (MR) was carried out in L-lysine-D-glucose (Lys-Glu) model system heated at 120°C for 0 to 10 h without pH control. Optical property (UV-Vis absorbance and fluorescence) development of MR was monitored. Antioxidant activity of maillard reaction products (MRPs) was investigated by a series of in vitro ...

  19. Lysine-Grafted MCM-41 Silica as an Antibacterial Biomaterial

    Science.gov (United States)

    Villegas, María F.; Garcia-Uriostegui, Lorena; Rodríguez, Ofelia; Izquierdo-Barba, Isabel; Salinas, Antonio J.; Toriz, Guillermo; Vallet-Regí, María; Delgado, Ezequiel

    2017-01-01

    This paper proposes a facile strategy for the zwitterionization of bioceramics that is based on the direct incorporation of l-lysine amino acid via the ε-amino group onto mesoporous MCM-41 materials. Fourier transform infrared (FTIR) studies of lysine-grafted MCM-41 (MCM-LYS) simultaneously showed bands at 3080 and 1540 cm−1 and bands at 1625 and 1415 cm−1 corresponding to -NH3+/COO− pairs, which demonstrate the incorporation of the amino acid on the material surface keeping its zwitterionic character. Both elemental and thermogravimetric analyses showed that the amount of grafted lysine was 8 wt. % based on the bioceramic total weight. Moreover, MCM-LYS exhibited a reduction of adhesion of S. aureus and E. coli bacteria in 33% and 50%, respectively at physiological pH, as compared with pristine MCM-41. Biofilm studies onto surfaces showed that lysine functionalization elicited a reduction of the area covered by S. aureus biofilm from 42% to only 5% (88%). This research shows a simple and effective approach to chemically modify bioceramics using single amino acids that provides zwitterionic functionality, which is useful to develop new biomaterials that are able to resist bacterial adhesion. PMID:28952559

  20. Effect of low protein diets and lysine supplementation on growth ...

    African Journals Online (AJOL)

    The present study was to assess the effect of feeding low protein diet with or without supplemental lysine to meet NRC (1998) requirement on growth performance, carcass trait, meat composition, and meat quality of pigs. An experiment of 126 days was conducted on 21 crossbred Landrace pigs (average weight 11.72 ...

  1. Enhancement of Monoclonal Antibody Production by Lysine-Containing Peptides

    Czech Academy of Sciences Publication Activity Database

    Franěk, František; Eckschlager, T.; Hermann, K.

    2003-01-01

    Roč. 19, č. 1 (2003), s. 169-174 ISSN 8756-7938 R&D Projects: GA MŠk OC 844.10 Institutional research plan: CEZ:AV0Z5038910; CEZ:MSM 111300005 Keywords : Monoclonal Antibody * Lysine-Containing Peptides Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 1.488, year: 2003

  2. Lysine metabolism in antisense C-hordein barley grains

    DEFF Research Database (Denmark)

    Schmidt, Daiana; Rizzi, Vanessa; Gaziola, Salete A

    2015-01-01

    The grain proteins of barley are deficient in lysine and threonine due to their low concentrations in the major storage protein class, the hordeins, especially in the C-hordein subgroup. Previously produced antisense C-hordein transgenic barley lines have an improved amino acid composition, with ...

  3. Detection of salt bridges to lysines in solution in barnase

    DEFF Research Database (Denmark)

    Hansen, Poul Erik; Williamson, Michael P.; Hounslow, Andrea M.

    2013-01-01

    We show that salt bridges involving lysines can be detected by deuterium isotope effects on NMR chemical shifts of the sidechain amine. Lys27 in the ribonuclease barnase is salt bridged, and mutation of Arg69 to Lys retains a partially buried salt bridge. The salt bridges are functionally important....

  4. Therapeutic use of chimeric bacteriophage (phage) lysins in staphylococcal endophthalmitis

    Science.gov (United States)

    Purpose: Phage endolysins are peptidoglycan hydrolases that are produced at the end of the phage lytic cycle to digest the host bacterial cell wall, facilitating the release of mature phage progeny. The aim of this study is to determine the antimicrobial activity of chimeric phage lysins against cli...

  5. Effect of Low Protein-Methionine-and-Lysine-Supplemented Diets ...

    African Journals Online (AJOL)

    Two experiments were conducted to investigate the effect of supplementing low CP diets with methionine and lysine on broiler performance, carcass measure and their immune response against Infectious Bursa Disease (IBD) virus. In Experiment 1, ten diets were formulated. Diet 1 (control diet) contained 23.0% CP and ...

  6. Identification of catechols as histone-lysine demethylase inhibitors

    DEFF Research Database (Denmark)

    Nielsen, Anders L; Kristensen, Line H; Stephansen, Karen B

    2012-01-01

    Identification of inhibitors of histone-lysine demethylase (HDM) enzymes is important because of their involvement in the development of cancer. An ELISA-based assay was developed for identification of inhibitors of the HDM KDM4C in a natural products library. Based on one of the hits with affinity...

  7. Effect Of Sprouting On Available Lysine Content Of Cowpea ( Vigna ...

    African Journals Online (AJOL)

    This study was conducted to determine the effect of sprouting on available Lysine content of cowpea (Vigna unguiculata) flour and the performance of the flour used for producing “moi – moi” (steamed bean cake). Cowpea seed was subjected to sprouting for different periods of 1 day, 2 days and 3 days for samples B, C and ...

  8. Lysine-Grafted MCM-41 Silica as an Antibacterial Biomaterial

    Directory of Open Access Journals (Sweden)

    María F. Villegas

    2017-09-01

    Full Text Available This paper proposes a facile strategy for the zwitterionization of bioceramics that is based on the direct incorporation of l-lysine amino acid via the ε-amino group onto mesoporous MCM-41 materials. Fourier transform infrared (FTIR studies of lysine-grafted MCM-41 (MCM-LYS simultaneously showed bands at 3080 and 1540 cm−1 and bands at 1625 and 1415 cm−1 corresponding to -NH3+/COO− pairs, which demonstrate the incorporation of the amino acid on the material surface keeping its zwitterionic character. Both elemental and thermogravimetric analyses showed that the amount of grafted lysine was 8 wt. % based on the bioceramic total weight. Moreover, MCM-LYS exhibited a reduction of adhesion of S. aureus and E. coli bacteria in 33% and 50%, respectively at physiological pH, as compared with pristine MCM-41. Biofilm studies onto surfaces showed that lysine functionalization elicited a reduction of the area covered by S. aureus biofilm from 42% to only 5% (88%. This research shows a simple and effective approach to chemically modify bioceramics using single amino acids that provides zwitterionic functionality, which is useful to develop new biomaterials that are able to resist bacterial adhesion.

  9. protein, tryptophan and lysine contents in quality protien maize

    African Journals Online (AJOL)

    owner

    study of protein, tryptophan and lysine composition of quality protein maize varieties (9). The tryptophan content of eleven superior QPM genotypes was much higher than those of wheat,. * Jimma University, College of Agriculture and Veterinary Medicine, Department of Crop Sciences, P.O. Box 307,. Jimma , Ethiopia.

  10. Growth responses to dietary lysine at high and low ambient temperatures in male turkeys

    NARCIS (Netherlands)

    Veldkamp, T.; Ferket, P.; Kwakkel, R.P.; Kogut, J.; Verstegen, M.W.A.

    2003-01-01

    Several researchers have postulated that dietary lysine requirements for turkeys are dependent upon ambient temperature. To test and quantify this hypothesis, a factorial experiment was designed with four dietary lysine levels (75, 90, 105, and 120% of NRC lysine recommendations) from 1 d of age

  11. Optimization of lysine production in Corynebacteriumglutamicum ATCC15032 by Response surface methodology

    Directory of Open Access Journals (Sweden)

    Mehrnaz Haghi

    2017-03-01

    Discussion and conclusion: According to the results, the proposed culture media by response surface methodology causes 1400 times increase in the lysine production compared with M9 culture media and methionine had an important role in the production of lysine, probably by inhibiting the other metabolic pathway which has common metabolic precursor with lysine production metabolic pathway.

  12. Microscopic analysis of MTT stained boar sperm cells

    Directory of Open Access Journals (Sweden)

    B.M. van den Berg

    2015-06-01

    Full Text Available The ability of sperm cells to develop colored formazan by reduction of MTT was used earlier to develop a spectrophotometric assay to determine the viability of sperm cells for several mammalian species. It was the objective of the present study to visualize microscopically the location of the formazan in boar sperm cells. The MTT staining process of boar sperm cells can be divided into a series of morphological events. Incubation of the sperm cells in the presence of MTT resulted after a few min in a diffuse staining of the midpiece of the sperm cells. Upon further incubation the staining of the midpiece became more intense, and gradually the formation of packed formazan granules became more visible. At the same time, a small formazan stained granule appeared medially on the sperm head, which increased in size during further incubation. After incubation for about 1 h the midpiece granules were intensely stained and more clearly distinct as granules, while aggregation of sperm cells occurred. Around 90% of the sperm cells showed these staining events. At the end of the staining the formazan granules have disappeared from both the sperm cells and medium, whereas formazan crystals appeared as thin crystal threads, that became heavily aggregated in the incubation medium. It was concluded that formazan is taken up by lipid droplets in the cytoplasm. Further, the use of the MTT assay to test for sperm viability should be regarded as a qualitative assay, whereas its practical use at artificial insemination (AI Stations is limited.

  13. Single bovine sperm sex typing by amelogenin nested PCR.

    Science.gov (United States)

    Colley, A; Buhr, M; Golovan, S P

    2008-10-01

    Sex-sorted bovine semen has become a valuable tool in animal production for sex preselection. Development of novel sperm sexing technologies, or evaluation of the quality of existing methods, often requires a single-sperm, sex-typing method that is reliable and easy to perform. In the present study, we report the development, validation, and application of a simple, reliable, and cost-effective method for single-sperm sex typing using nested polymerase chain reaction (PCR), based on the amelogenin gene. Several hundred single sperm were isolated using a simple manual technique, or a high-speed flow-sorter, and were successfully sex-typed using the amelogenin nested PCR. Based on the pooled results of individual sperm, there was no significant difference in the semen sex ratio of unsorted (44.6% X-sperm and 55.4% Y-sperm) or X/Y-sorted semen (91.4% X-sperm and 94.0% Y-sperm), as compared to the expected ratio in unsorted semen or the post-sorting reanalysis data, respectively. The amelogenin single-sperm sexing method was an adaptable, accurate, and reliable tool for single-sperm sex typing.

  14. Increased competitiveness of nematode sperm bearing the male X chromosome.

    Science.gov (United States)

    LaMunyon, C W; Ward, S

    1997-01-07

    Male offspring, which cannot reproduce independently, represent a cost of sexual reproduction. This cost is eliminated by the production of hermaphroditic offspring in the self-fertilizing nematode Caenorhabditis briggsae. However, these hermaphrodites can outcross by mating with males. Half the sperm received from males contain no sex chromosome and therefore give rise to male progeny. Mating with males should thus impose the cost of making male offspring. We found that male sperm took immediate precedence over hermaphrodite sperm, resulting in maximized outcrossing, but the appearance of male progeny was delayed after mating. This delay is caused by the male X-bearing sperm outcompeting their nullo-X counterparts. The competitive advantage of X-bearing sperm over nullo-X sperm is limited to sperm from males; it did not occur in a mutant hermaphrodite that produces both types of sperm. The chromosomal effect on sperm competitiveness in C. briggsae, which has not been observed in other species, suggests that the X chromosome has evolved a form of meiotic drive, selfishly increasing the competitiveness of sperm that bear it over those that do not. Thus, the multiple levels of sperm competitiveness found in C. briggsae maximize outcrossing after mating while delaying the cost of making male offspring.

  15. The sperm of Hylodinae species (Anura, Leptodactylidae ...

    Indian Academy of Sciences (India)

    Home; Journals; Journal of Biosciences; Volume 31; Issue 3. The sperm of Hylodinae species (Anura, Leptodactylidae): Ultrastructural characteristics and their relevance to interspecific taxonomic relationships. O Aguiar-Jr A A Giaretta S M Recco-Pimentel. Articles Volume 31 Issue 3 September 2006 pp 379-388 ...

  16. Comparative evaluation of the sperm characteristics and ...

    African Journals Online (AJOL)

    Comparative evaluation of the sperm characteristics and morphology of adult Wistar rats fed either low or normal protein-energy diets and orally dosed with ... of adult Wistar rats orally dosed aqueous extracts of C. australis seed (LPSE and NPSE) and stem (LPST and NPST), 300mg of extract/kg body weight of rat/day, ...

  17. Exposures that may affect sperm DNA integrity

    DEFF Research Database (Denmark)

    Håkonsen, L B; Spano, M; Bonde, J P

    2012-01-01

    Prenatal lifestyle exposures are linked to alterations in conventional semen characteristics. Sperm DNA integrity is another marker of semen quality shown to be altered in mice prenatally exposed to chemicals. From a Danish pregnancy cohort established in 1984-1987, sons were selected for a follow...

  18. Comparative evaluation of the sperm characteristics and ...

    African Journals Online (AJOL)

    olayemitoyin

    Comparative evaluation of the sperm characteristics and morphology of adult Wistar rats fed either low or normal protein-energy diets and orally dosed with aqueous Cuscuta australis extracts. 1. Omirinde, J.O., *. 1. Ozegbe, P.C. and. 2. Oyeyemi, M.O.. Departments of 1Veterinary Anatomy and 2Veterinary Surgery and ...

  19. Psychosocial counselling in donor sperm treatment

    NARCIS (Netherlands)

    Visser, M.

    2018-01-01

    For decades, donor sperm treatment is offered to men and women to build a family. In daily life, parents, children and donors have to deal with the consequences of this treatment. The studies of this thesis show that there are gaps in knowledge about specialist psychosocial counselling and guidance

  20. Cryopreservation of mutton snapper ( Lutjanus analis sperm

    Directory of Open Access Journals (Sweden)

    EDUARDO G. SANCHES

    2013-09-01

    Full Text Available This study aimed to develop a protocol of semen cryopreservation of the mutton snapper Lutjanus analis. The interaction between three extenders ( pH 6.1; 7.8 and 8.2 , two concentrations of dimethyl sulfoxide ( DMSO, 5 and 10% and three cooling rates ( -90; -60 and -30°C.min−1 on the sperm motility rate and motility time were analyzed by a factorial experiment. A sample of 30 fishes ( 1,261 ± 449 g collected in the nature was kept in floating net cages. The semen was frozen by using cryogenic straws, in nitrogen vapour and transferred, later, to liquid nitrogen. Fertilization test was accomplished to evaluate the viability of the cryopreserved sperm. The highest sperm motility rate and motility time ( P < 0.05 was achieved by combining extender C ( pH 8.2 with DMSO ( 10% and cooling rate of -60°C.min−1 ( P < 0.05 . The use of cryopreserved sperm presented fertilization rates higher than 59% validating the present protocol for mutton snapper.

  1. Sperm Quality and Fertility Following Prolonged Immobilization ...

    African Journals Online (AJOL)

    Sperm Quality and Fertility Following Prolonged Immobilization Stress in Wistar Rats. ... All the females that were mated with the control males became pregnant and produced litters. On the ... These results therefore, confirm and expand existing literatures reporting impaired fertility following long-term exposure to stress.

  2. The relationship between acridine orange fluorescence of sperm nuclei and the fertilizing ability of human sperm.

    Science.gov (United States)

    Hoshi, K; Katayose, H; Yanagida, K; Kimura, Y; Sato, A

    1996-10-01

    To determine whether the outcome of IVF can be predicted by acridine orange (AO) nuclear fluorescence of sperm. Based on the fact that AO nuclear fluorescence color after acid treatment reflects maturity (green fluorescence) or immaturity (yellow to red fluorescence) of spermatozoa, the relationships between sperm maturity and the outcome of IVF, subzonal insemination, or intracytoplasmic sperm injection (ICSI) were investigated. The IVF program at the Obstetrics and Gynecology Hospital, Fukushima Medical College. Sixty-eight patients undergoing 68 IVF treatment cycles. Acridine orange fluorescence of sperm nuclei and successful oocyte fertilization. conventional semen parameters (sperm concentration and percentages of motile or morphologically normal spermatozoa in semen) did not correlate with the incidence of spermatozoa with green AO fluorescent (mature) nuclei. When > or = 50% of spermatozoa in semen samples exhibited green AO nuclear fluorescence, IVF was always successful. When green AO nuclear fluorescence was green AO fluorescent spermatozoa were able to bind efficiently human zona pellucida. When the incidence of green AO fluorescent spermatozoa was fertilized by ICSI. The spermatozoa which fertilized oocytes in vivo and in IVF were limited to those whose nuclei exhibited green AO fluorescence. Intracytoplasmic sperm injection may be the method of choice when the incidence of green AO nuclear fluorescence is low regardless of the results of semen analysis.

  3. Microsurgical and Percutaneous Epididymal Sperm Aspiration for Sperm Collection from Live Mice.

    Science.gov (United States)

    Boersma, Auke; Olszanska, Olga; Walter, Ingrid; Rülicke, Thomas

    2015-09-01

    Spermatozoa for in vitro fertilization of mouse oocytes and other methods of assisted reproduction typically are collected from the cauda epididymis of euthanized male mice. As an alternative to this terminal protocol, we developed and examined 2 methods for collecting sperm from anesthetized male mice without decreasing subsequent fertility: microsurgical epididymal sperm aspiration and, as a refinement, percutaneous epididymal sperm aspiration. Collected sperm was evaluated in terms of motility, concentration and in vitro fertilization ability. After recovery, both treated and untreated control male mice underwent in vivo fertility testing and subsequent histologic analysis of the treated male reproductive tract (epididymis and testis). In vitro fertilization using sperm recovered by the 2 collection methods was successfully achieved in all cases. The in vivo fertility test and the histologic analysis revealed no impairment of fertility and no permanent histologic alteration in the treated mice. Therefore, we recommend both techniques as simple and effective methods for recovering high-quality epididymal mouse sperm without having to euthanize fertile male mice.

  4. Effect of environmental pH on sperm kinematic characteristics

    DEFF Research Database (Denmark)

    Alipour, Hiva; Dardmeh, Fereshteh; Dissing, Marianne Charlotte

    2015-01-01

    Semen preparation medium have an important role in assisted reproduction techniques and their composition influences sperm binding and motility. Some studies have assessed the influence of pH on sperm kinetics. However, no study to date has assessed the effect of environmental pH on subtle...... differences in the details of the sperm movement (kinematics) of human sperm provided by computerized sperm analysis systems. This study was designed to assess the effect of two different media pH levels on kinematic parameters of the human sperm. Samples were prepared using the 40%/80% Pureception (Sage, USA....... Linearity and Wobble showed significant difference after 24 hours. This study demonstrated a difference in the sperm motion pattern and velocity in different environmental pH levels. Based on these findings, further investigations are required to elucidate knowledge about possible effect of marginal p...

  5. Sperm cells as vectors in the production of transgenic animals

    Energy Technology Data Exchange (ETDEWEB)

    Prince, R.M.

    1993-04-28

    Transgenic animals are used in industry and in biomedical research in order to provide in vivo experimental model systems. Sperm cells have been reported used as vectors in the production of transgenic animals before, however no approach has of yet proven to be successful. Fertilizing eggs with genetically modified sperm would be advantageous in that sperm are readily accessible and stable, and eggs can be fertilized by modified sperm cells in vivo. Recent elucidations regarding the unique manner of DNA packaging in sperm chromatin by protamines has provided us with the insight for developing a method of introducing foreign DNA into sperm which is likely to succeed where others have failed. We have developed a method for mimicking the in vivo system of sperm chromatin toroid subunits in vitro, concentrating these toroids, and fluorescent visualization. Our present work concerns development of a method to successfully deliver DNA across the cell membranes and into the nucleus.

  6. Left-handed sperm removal by male Calopteryx damselflies (Odonata).

    Science.gov (United States)

    Tsuchiya, Kaori; Hayashi, Fumio

    2014-01-01

    Male genitalia in several insect species are asymmetry in right and left shape. However, the function of such asymmetric male genitalia is still unclear. We found that the male genitalia of the damselfly Calopteryx cornelia (Odonata: Calopterygidae) are morphologically symmetric just after emergence but asymmetric after reproductive maturation. Males remove rival sperm stored in the female bursa copulatrix (single spherical sac) and the following spermatheca (Y-shaped tubular sac) prior to their own ejaculation to prevent sperm competition. Males possess the aedeagus with a recurved head to remove bursal sperm and a pair of spiny lateral processes to remove spermathecal sperm. The right lateral process is less developed than the left, and sperm stored in the right spermathecal tube are rarely removed. Experiments involving surgical cutting of each lateral process demonstrated that only the left process functions in spermathecal sperm removal. Thus, males of C. cornelia are left-handed in their sperm removal behaviour at copulation.

  7. Response of midpiece vesicles on human sperm to osmotic stress

    DEFF Research Database (Denmark)

    Abraham-Peskir, Joanna V; Chantler, Eric; Uggerhøj, Erik

    2002-01-01

    BACKGROUND: We investigated the osmotic response of midpiece vesicles (MPV) on human sperm. METHODS: Light microscopy, transmission X-ray microscopy and computer-aided semen analysis was used to investigate sperm in normozoospermic semen from healthy donors, separated from semen and suspended...... in hyper- or hypo-osmotic solutions. RESULTS: The mean incidence of MPV-bearing sperm in semen was 31% (range 8-71; n = 47). MPV were morphologically different from cytoplasmic droplets. The incidence of MPV-bearing sperm in separated populations increased reversibly after washing in Sperm Preparation...... Medium but not after washing in seminal plasma. There was an inverse relationship between medium osmolality and both MPV-bearing sperm incidence and MPV diameter. However, initial osmolality in semen from different donors did not correlate with incidence of MPV-bearing sperm. Furthermore, a direct...

  8. Sperm storage induces an immunity cost in ants

    DEFF Research Database (Denmark)

    Baer, Boris; Armitage, Sophie A O; Boomsma, Jacobus J

    2006-01-01

    Ant queens are among the most long-lived insects known. They mate early in adult life and maintain millions of viable sperm in their sperm storage organ until they die many years later. Because they never re-mate, the reproductive success of queens is ultimately sperm-limited, but it is not known...... what selective forces determine the upper limit to sperm storage. Here we show that sperm storage carries a significant cost of reduced immunity during colony founding. Newly mated queens of the leaf-cutting ant Atta colombica upregulate their immune response shortly after completing their nest burrow......, probably as an adaptive response to a greater exposure to pathogens in the absence of grooming workers. However, the immune response nine days after colony founding is negatively correlated with the amount of sperm in the sperm storage organ, indicating that short-term survival is traded off against long...

  9. COMP-1 promotes competitive advantage of nematode sperm.

    Science.gov (United States)

    Hansen, Jody M; Chavez, Daniela R; Stanfield, Gillian M

    2015-03-19

    Competition among sperm to fertilize oocytes is a ubiquitous feature of sexual reproduction as well as a profoundly important aspect of sexual selection. However, little is known about the cellular mechanisms sperm use to gain competitive advantage or how these mechanisms are regulated genetically. In this study, we utilize a forward genetic screen in Caenorhabditis elegans to identify a gene, comp-1, whose function is specifically required in competitive contexts. We show that comp-1 functions in sperm to modulate their migration through and localization within the reproductive tract, thereby promoting their access to oocytes. Contrary to previously described models, comp-1 mutant sperm show no defects in size or velocity, thereby defining a novel pathway for preferential usage. Our results indicate not only that sperm functional traits can influence the outcome of sperm competition, but also that these traits can be modulated in a context-dependent manner depending on the presence of competing sperm.

  10. The biology of lysine acetylation integrates transcriptional programming and metabolism

    Directory of Open Access Journals (Sweden)

    Mujtaba Shiraz

    2011-03-01

    Full Text Available Abstract The biochemical landscape of lysine acetylation has expanded from a small number of proteins in the nucleus to a multitude of proteins in the cytoplasm. Since the first report confirming acetylation of the tumor suppressor protein p53 by a lysine acetyltransferase (KAT, there has been a surge in the identification of new, non-histone targets of KATs. Added to the known substrates of KATs are metabolic enzymes, cytoskeletal proteins, molecular chaperones, ribosomal proteins and nuclear import factors. Emerging studies demonstrate that no fewer than 2000 proteins in any particular cell type may undergo lysine acetylation. As described in this review, our analyses of cellular acetylated proteins using DAVID 6.7 bioinformatics resources have facilitated organization of acetylated proteins into functional clusters integral to cell signaling, the stress response, proteolysis, apoptosis, metabolism, and neuronal development. In addition, these clusters also depict association of acetylated proteins with human diseases. These findings not only support lysine acetylation as a widespread cellular phenomenon, but also impel questions to clarify the underlying molecular and cellular mechanisms governing target selectivity by KATs. Present challenges are to understand the molecular basis for the overlapping roles of KAT-containing co-activators, to differentiate between global versus dynamic acetylation marks, and to elucidate the physiological roles of acetylated proteins in biochemical pathways. In addition to discussing the cellular 'acetylome', a focus of this work is to present the widespread and dynamic nature of lysine acetylation and highlight the nexus that exists between epigenetic-directed transcriptional regulation and metabolism.

  11. Sperm flagellum volume determines freezability in red deer spermatozoa.

    Directory of Open Access Journals (Sweden)

    José Luis Ros-Santaella

    Full Text Available The factors affecting the inter-individual differences in sperm freezability is a major line of research in spermatology. Poor sperm freezability is mainly characterised by a low sperm velocity, which in turn is associated with low fertility rates in most animal species. Studies concerning the implications of sperm morphometry on freezability are quite limited, and most of them are based on sperm head size regardless of the structural parts of the flagellum, which provides sperm motility. Here, for the first time, we determined the volumes of the flagellum structures in fresh epididymal red deer spermatozoa using a stereological method under phase contrast microscopy. Sperm samples from thirty-three stags were frozen and classified as good freezers (GF or bad freezers (BF at two hours post-thawing using three sperm kinetic parameters which are strongly correlated with fertility in this species. Fourteen stags were clearly identified as GF, whereas nineteen were BF. No significant difference in sperm head size between the two groups was found. On the contrary, the GF exhibited a lower principal piece volume than the BF (6.13 µm3 vs 6.61 µm3, respectively, p = 0.006. The volume of the flagellum structures showed a strong negative relationship with post-thawing sperm velocity. For instance, the volume of the sperm principal piece was negatively correlated with sperm velocity at two hours post-thawing (r = -0.60; p<0.001. Our results clearly show that a higher volume of the sperm principal piece results in poor freezability, and highlights the key role of flagellum size in sperm cryopreservation success.

  12. Sperm antibody production in female sterility.

    Science.gov (United States)

    Mettler, L; Scheidel, P; Shirwani, D

    1974-01-01

    A review of the immunological implications in reproductive physiology is presented. Although attempts have been made to ascribe the antigenicity of semen to individual components, it has not been possible to isolate the human semen antigen responsible for infertility. In monkeys total ejaculates and seminal plasma have shown higher antigenicity than washed spermatozoa. In bulls some evidence of such antigens have been found in the seminal plasma. They are iron-binding proteins resembling lactoferrin. Most investigators have found no evidence for any participation of the ABO blood group antigens in cases of sterility. On the surface of human spermatozoa histo-incompatibility antigens have been detected. Transplantation antigens may be related to sterility. However, an immulogic tolerance of the maternal organism exists against the genetically foreign fetal tissue. Autoimmune spermagglutinating antibodies have been detected in the sera and in the seminal plasma of males with sterility. An obstruction of the seminal pathways may facilitate the production of such antibodies against retained sperm. Isoimmunity in females against seminal components has been shown in cases of sterility; however, fertile women have also been shown to have such conditions. In a group of infertile women spermagglutination activity was detected in 7.5% of cases. In another series of 46 cases with primary unexplained infertility agglutinating antibodies were found in 17.4%. Other investigators have also reported higher rates than the authors. The sperm immobilization test seems to be more sensitive than the agglutination test. No sera were found positive with both tests. With immunofluorescent techniques humoral sperm antibodies have been found to be the IgM and IgG fractions. Each acts on a different part of the spermatozoa. The only promising therapy against humoral sperm antibodies is avoidance of sperm contact over a long period of time. Reported results have been conflicting. Cortisone

  13. Genomic characterization and expression profiles upon bacterial infection of a novel cystatin B homologue from disk abalone (Haliotis discus discus).

    Science.gov (United States)

    Premachandra, H K A; Wan, Qiang; Elvitigala, Don Anushka Sandaruwan; De Zoysa, Mahanama; Choi, Cheol Young; Whang, Ilson; Lee, Jehee

    2012-12-01

    Cystatins are a large family of cysteine proteinase inhibitors which are involved in diverse biological and pathological processes. In the present study, we identified a gene related to cystatin superfamily, AbCyt B, from disk abalone Haliotis discus discus by expressed sequence tag (EST) analysis and BAC library screening. The complete cDNA sequence of AbCyt B is comprised of 1967 nucleotides with a 306 bp open reading frame (ORF) encoding for 101 amino acids. The amino acid sequence consists of a single cystatin-like domain, which has a cysteine proteinase inhibitor signature, a conserved Gly in N-terminal region, QVVAG motif and a variant of PW motif. No signal peptide, disulfide bonds or carbohydrate side chains were identified. Analysis of deduced amino acid sequence revealed that AbCyt B shares up to 44.7% identity and 65.7% similarity with the cystatin B genes from other organisms. The genomic sequence of AbCyt B is approximately 8.4 Kb, consisting of three exons and two introns. Phylogenetic tree analysis showed that AbCyt B was closely related to the cystatin B from pacific oyster (Crassostrea gigas) under the family 1.Functional analysis of recombinant AbCyt B protein exhibited inhibitory activity against the papain, with almost 84% inhibition at a concentration of 3.5 μmol/L. In tissue expression analysis, AbCyt B transcripts were expressed abundantly in the hemocyte, gill, mantle, and digestive tract, while weakly in muscle, testis, and hepatopancreas. After the immune challenge with Vibrio parahemolyticus, the AbCyt B showed significant (P<0.05) up-regulation of relative mRNA expression in gill and hemocytes at 24 and 6 h of post infection, respectively. These results collectively suggest that AbCyst B is a potent inhibitor of cysteine proteinases and is also potentially involved in immune responses against invading bacterial pathogens in abalone. Copyright © 2012 Elsevier Ltd. All rights reserved.

  14. A new approach to sperm preservation based on bioenergetic theory.

    Science.gov (United States)

    Froman, D P; Feltmann, A J

    2010-04-01

    To date, attempts to preserve chicken sperm have been based on a trial-and-error experimental approach. The present work outlines the development of an alternative approach based on empiricism and bioenergetic theory. In previous work, we found fowl sperm motility to be dependent on mitochondrial calcium cycling, phospholipase A(2), and long-chain fatty acids as an endogenous energy source. It is noteworthy that fowl sperm reside within the sperm storage tubules (SST) of the oviduct over an interval of days to weeks after insemination. In this regard, a model for in vivo sperm storage was developed and tested in additional previous research. Sperm penetration of the SST, sperm residence within the SST, and sperm egress from the SST can be explained in terms mitochondrial function. Understanding sperm function and longevity in terms of bioenergetics presented the possibility that sperm could be inactivated by disrupting mitochondrial calcium cycling and could thereby be preserved. However, this possibility also posed a problem: maintenance of the inner membrane potential of the mitochondrion within inactivated sperm. This report describes a series of experiments in which fowl sperm were inactivated by treatment with the calcium chelator tetrasodium 1,2-bis-(o-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid, and then reactivated by treatment with calcium ions. The effect of tetrasodium 1,2-bis-(o-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid on mitochondrial calcium cycling was confirmed by flow cytometry and confocal microscopy. When treated sperm were cooled to 10 degrees C, inactivated sperm could be reactivated throughout a 5-h storage interval. When stored sperm were held for 3 h before reactivation and insemination, fertility was 88% of the control. Storage did not affect hatchability. In summary, short-term storage was realized by manipulating mitochondrial function. We propose that 1) complex V consumes ATP within inactivated sperm and, by doing so, maintains

  15. Effects of cryopreservation on sperm viability, synthesis of reactive oxygen species, and DNA damage of bovine sperm.

    Science.gov (United States)

    Gürler, H; Malama, E; Heppelmann, M; Calisici, O; Leiding, C; Kastelic, J P; Bollwein, H

    2016-07-15

    The objective was to examine if there are relationships between alterations in sperm viability, reactive oxygen species (ROS) synthesis, and DNA integrity induced by cryopreservation of bovine sperm. Four ejaculates were collected from each of six bulls. Each ejaculate was diluted and divided into two aliquots; one was incubated for 24 hours at 37 °C, and the other frozen, thawed, and incubated for 24 hours at 37 °C. Analyses of quality of sperm were performed after 0, 3, 6, 12, and 24 hours of incubation. Progressive motile sperm was determined with computer assisted sperm analysis. Percentages of plasma membrane- and acrosome-intact sperm, sperm with a high mitochondrial membrane potential, sperm showing a high degree of DNA fragmentation (%DFI), and their reactive oxygen species content were assessed with dichlorofluorescein-diacetate, dihydrorhodamine, diaminofluorescein diacetate, and mitochondrial superoxide indicator using flow cytometry. Although all other sperm parameters showed alterations (P  0.05, 0.91 ± 0.23) in nonfrozen sperm. Cryopreservation induced changes of all sperm parameters (P reactive oxygen species. Copyright © 2016 Elsevier Inc. All rights reserved.

  16. Assessment of sperm DNA fragmentation in stallion (Equus caballus) and donkey (Equus asinus) using the sperm chromatin dispersion test.

    Science.gov (United States)

    Cortés-Gutiérrez, E I; Crespo, F; Serres-Dalmau, C; Gutiérrez de las Rozas, A L; Dávila-Rodríguez, M I; López-Fernández, C; Gósalvez, J

    2009-10-01

    Sperm DNA fragmentation (sDF) is an important parameter to assessing sperm quality. Information about sperm quality is not available for donkeys, especially in some breeds at risk of extinction. The objectives of this research were to test the four commercial variants of sperm chromatin dispersion test (SCD; sperm Halomax test), originally developed to assess sDF in boars, bulls, rams and stallions, in order to scrutinize their applicability in the study of sDF in a donkey breed at risk of extinction (Zamorano-Leonesa), for which there is no specific test available to analyze sperm at present. Only the SCD test, originally developed for stallions, produced stable and consistent results, and was deemed suitable to assess DNA fragmentation in sperm samples from donkeys. Image analysis was used to compare differences between the SCD methodology applied to stallion and donkey semen samples processed under the same experimental conditions. The extent of SCD in the SCD test was approximately 20% lower in donkey sperm than in stallion sperm. Yet, the ratio of chromatin sperm dispersion achieved in fragmented and unfragmented nuclei did not differ significantly between species. These data suggest that a similar protein depletion treatment can cause differences in protein removal in equivalent cells from different species and that sperm chromatin may be organized differently in stallions and donkeys.

  17. Motile sperm organelle morphology examination (MSOME: intervariation study of normal sperm and sperm with large nuclear vacuoles

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    Baruffi Ricardo LR

    2010-06-01

    Full Text Available Abstract Background Although the motile sperm organelle morphology examination (MSOME was developed only as a selection criterion, its application as a method for classifying sperm morphology may represent an improvement in evaluation of semen quality, with potential clinical repercussions. The present study aimed to evaluate individual variations in the motile sperm organelle morphology examination (MSOME analysis after a time interval. Methods Two semen samples were obtained from 240 men from an unselected group of couples undergoing infertility investigation and treatment. Mean time interval between the two semen evaluations was 119 +/- 102 days. No clinical or surgical treatment was realized between the two observations. Spermatozoa were analyzed at greater than or equal to 8400× magnification by inverted microscope equipped with DIC/Nomarski differential interference contrast optics. At least 200 motile spermatozoa per semen sample were evaluated and percentages of normal spermatozoa and spermatozoa with large nuclear vacuoles (LNV/one or more vacuoles occupying >50% of the sperm nuclear area were determined. A spermatozoon was classified as morphologically normal when it exhibited a normal nucleus (smooth, symmetric and oval nucleus, width 3.28 +/- 0.20 μm, length 4.75 +/- 0.20 μm/absence of vacuoles occupying >4% of nuclear area as well as acrosome, post-acrosomal lamina, neck and tail, besides not presenting cytoplasm around the head. One examiner, blinded to subject identity, performed the entire study. Results Mean percentages of morphologically normal and LNV spermatozoa were identical in the two MSOME analyses (1.6 +/- 2.2% vs. 1.6 +/- 2.1% P = 0.83 and 25.2 +/- 19.2% vs. 26.1 +/- 19.0% P = 0.31, respectively. Regression analysis between the two samples revealed significant positive correlation for morphologically normal and for LNV spermatozoa (r = 0.57 95% CI:0.47-0.65 P P Conclusions The significant positive correlation and

  18. Decline of semen quality among Chinese sperm bank donors within 7 years (2008-2014

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    Li Wang

    2017-01-01

    Full Text Available Semen from 5210 sperm bank donors was analyzed and trends in semen quality were evaluated at Shandong Human Sperm Bank between 2008 and 2014. After 2-7 days of abstinence, semen samples were collected. Measurements of semen volume, sperm concentration, sperm forward motility, and total sperm count were performed. There were significant declining trends in semen volume, sperm concentration, sperm forward motility, and total sperm count. Our results indicate that the quality of semen in this cohort of sperm donors had decreased during the study period.

  19. Sperm morphology and function in passerine birds: insights from intra- and interspecific studies

    OpenAIRE

    Laskemoen, Terje

    2010-01-01

    Sperm cells are the most variable animal cells, and a tremendous variation in sperm phenotypes exists among species, from minute amoeboid sperm to giant sperm in some species of fruit flies. Much effort has been devoted to the study of sperm evolution. Sperm competition, when sperm from two or more males compete over fertilization of a set of ova, is thought to be the major force driving the evolution of sperm. There are several ways in which sperm competition can effect the evolution of sper...

  20. Human sperm head vacuoles are physiological structures formed during the sperm development and maturation process.

    Science.gov (United States)

    Tanaka, Atsushi; Nagayoshi, Motoi; Tanaka, Izumi; Kusunoki, Hiroshi

    2012-08-01

    To clarify whether human sperm vacuoles affected intracytoplasmic sperm injection (ICSI) success rates. Retrospective study. A private infertility clinic. Spermatozoa and spermatids were obtained from 11 normozoospermic, 10 oligozoospermic or asthenozoospermic, 4 obstructive azoospermic, and 3 nonobstructive azoospermic men. Differential interference contrast observation and intracytoplasmic injection of morphologically selected sperm. Incidence, size, and position of vacuoles of sperm cells were recorded. Ability of fertilization and blastocyst development were compared between cells with and without vacuoles. More than 97.4% of ejaculated, 87.5% of epididymal, 87.5% of testicular spermatozoa, and more than 90.0% of Sc-Sd2 spermatids had vacuoles of various sizes. The incidence of vacuoles on ejaculated cells was significantly higher than that on the other types of cells, but there was no difference between sperm from normozoospermic men and those from the other donors. Removal of plasma membrane and/or acrosome did not affect the incidence of vacuoles. Although more than 60% of spermatozoa had small vacuoles in the acrosomal regions, 52.6% of Sb1-2 spermatids had large vacuoles. After injection of a motile spermatozoon with large and small vacuoles, 60.9% and 85.7% of metaphase II oocytes could be normally fertilized, respectively, and almost half of the zygotes developed to the blastocyst stage. When using sperm without vacuoles, the fertilization rate was 80.0%, but only 25% of them developed to the blastocyst stage. Human sperm head vacuoles did not affect ICSI outcomes. Crown Copyright © 2012. Published by Elsevier Inc. All rights reserved.

  1. Effect of hydroxylysine on the biosynthesis of lysine in Streptococcus faecalis.

    Science.gov (United States)

    Gilboe, D P; Friede, J D; Henderson, L M

    1968-03-01

    We were able to show that two lysine-independent mutants of Streptococcus faecalis ATCC 8043 contained the enzymes for the usual bacterial pathway for lysine biosynthesis. Because of this synthetic capacity, one mutant, the Lys(+)OHLys(s) strain, could not grow in the presence of hydroxylysine without a lysine supplement. Both lysine and hydroxylysine inhibited the first enzyme of the pathway, aspartokinase. Unlike the Escherichia coli enzyme, S. faecalis dihydrodipicolinic acid synthetase was not inhibited by either lysine or hydroxylysine. Both amino acids caused the repression of dihydrodipicolinic acid synthetase and diaminopimelic acid decarboxylase. Failure of Lys(+)OHLys(s) strain to grow in hydroxylysine-supplemented medium was caused by the mimicking of lysine control by hydroxylysine. Because hydroxylysine could not completely substitute for lysine and lysine could not be synthesized, the organism did not grow. We tested three lysine analogues and found that they prevented lysine-depletion lysis in the Lsy(-)OHLys(s) strain, as did hydroxylysine. Each analogue seemed to support cell wall mucopeptide synthesis, although ornithine did not. Preliminary data indicated that these analogues like hydroxylysine, have growth-inhibitory action on the Lys(+)OHLys(s) strain, but not the Lys(+)OHLys(r) strain. The nature of the specificity of the lysine-adding enzyme for cell wall mucopeptide synthesis is discussed.

  2. Sperm vacuoles are linked to capacitation and acrosomal status.

    Science.gov (United States)

    Montjean, Debbie; Belloc, Stephanie; Benkhalifa, Moncef; Dalleac, Alain; Ménézo, Yves

    2012-10-01

    Is the presence of nuclear vacuoles really a negative parameter? As sperm vacuoles are associated with acrosomal and capacitation status, they appear to be a reflection of normal sperm physiology. The selection of sperm under a high magnification has been proposed as a strategy to increase the success rates of ICSI, through a better selection of sperm for injection. The presence of vacuoles on the sperm head is said to be a negative parameter. We incubated processed sperm for 90 min with two strong inducers of acrosome reaction (AR), i.e. hyaluronic acid (HA) and follicular fluid (FF) and studied the evolution of nuclear vacuoles, sperm morphology and chromatin compaction. We tested the effect of incubating sperm samples with HA and FF for 90 min at 37°C on nuclear vacuoles. Both HA and FF strongly induce AR after 90 min, without significantly modifying sperm nuclear condensation and morphology (Bartoov's criteria). We simultaneously observed a highly significant decrease in the presence of vacuoles. This is a descriptive study based on in vitro manipulations. Although intracytoplasmic morphologically selected sperm injection may be of benefit for couples with specific treatment indications, the results of this study make it difficult to justify its large-scale application. Funding was granted by Laboratoire d'Eylau, Unilabs.

  3. Factors affecting sperm quality before and after mating of calopterygid damselflies.

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    Kaori Tsuchiya

    Full Text Available Damselflies (Odonata: Zygoptera have a more complex sperm transfer system than other internally ejaculating insects. Males translocate sperm from the internal reproductive organs to the specific sperm vesicles, a small cavity on the body surface, and then transfer them into the female. To examine how the additional steps of sperm transfer contribute to decreases in sperm quality, we assessed sperm viability (the proportion of live sperm at each stage of mating and after different storage times in male and female reproductive organs in two damselfly species, Mnais pruinosa and Calopteryx cornelia. Viability of stored sperm in females was lower than that of male stores even just after copulation. Male sperm vesicles were not equipped to maintain sperm quality for longer periods than the internal reproductive organs. However, the sperm vesicles were only used for short-term storage; therefore, this process appeared unlikely to reduce sperm viability when transferred to the female. Males remove rival sperm prior to transfer of their own ejaculate using a peculiar-shaped aedeagus, but sperm removal by males is not always complete. Thus, dilution occurs between newly received sperm and aged sperm already stored in the female, causing lower viability of sperm inside the female than that of sperm transferred by males. If females do not remate, sperm viability gradually decreases with the duration of storage. Frequent mating of females may therefore contribute to the maintenance of high sperm quality.

  4. The Semen pH Affects Sperm Motility and Capacitation.

    Science.gov (United States)

    Zhou, Ji; Chen, Li; Li, Jie; Li, Hongjun; Hong, Zhiwei; Xie, Min; Chen, Shengrong; Yao, Bing

    2015-01-01

    As the chemical environment of semen can have a profound effect on sperm quality, we examined the effect of pH on the motility, viability and capacitation of human sperm. The sperm in this study was collected from healthy males to avoid interference from other factors. The spermatozoa cultured in sperm nutrition solution at pH 5.2, 6.2, 7.2 and 8.2 were analyzed for sperm total motility, progressive motility (PR), hypo-osmotic swelling (HOS) rate, and sperm penetration. Our results showed that these parameters were similar in pH 7.2 and 8.2 sperm nutrition solutions, but decreased in pH 5.2 and 6.2 solutions. The HOS rate exhibited positive correlation with the sperm total motility and PR. In addition, the sperm Na(+)/K(+)-ATPase activity at different pHs was measured, and the enzyme activity was significantly lower in pH 5.2 and 6.2 media, comparing with that in pH 8.2 and pH 7.2 solutions. Using flow cytometry (FCM) and laser confocal scanning microscopy (LCSM) analysis, the intracellular Ca2(+ )concentrations of sperm cultured in sperm capacitation solution at pH 5.2, 6.2, 7.2 and 8.2 were determined. Compared with that at pH 7.2, the mean fluorescence intensity of sperm in pH 5.2 and 6.2 media decreased significantly, while that of pH 8.2 group showed no difference. Our results suggested that the declined Na(+)/K(+)-ATPase activity at acidic pHs result in decreased sperm movement and capacitation, which could be one of the mechanisms of male infertility.

  5. DNA fragmentation and sperm head morphometry in cat epididymal spermatozoa.

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    Vernocchi, Valentina; Morselli, Maria Giorgia; Lange Consiglio, Anna; Faustini, Massimo; Luvoni, Gaia Cecilia

    2014-10-15

    Sperm DNA fragmentation is an important parameter to assess sperm quality and can be a putative fertility predictor. Because the sperm head consists almost entirely of DNA, subtle differences in sperm head morphometry might be related to DNA status. Several techniques are available to analyze sperm DNA fragmentation, but they are labor-intensive and require expensive instrumentations. Recently, a kit (Sperm-Halomax) based on the sperm chromatin dispersion test and developed for spermatozoa of different species, but not for cat spermatozoa, became commercially available. The first aim of the present study was to verify the suitability of Sperm-Halomax assay, specifically developed for canine semen, for the evaluation of DNA fragmentation of epididymal cat spermatozoa. For this purpose, DNA fragmentation indexes (DFIs) obtained with Sperm-Halomax and terminal deoxynucleotidyl transferase-mediated nick-end labeling (TUNEL) were compared. The second aim was to investigate whether a correlation between DNA status, sperm head morphology, and morphometry assessed by computer-assisted semen analysis exists in cat epididymal spermatozoa. No differences were observed in DFIs obtained with Sperm-Halomax and TUNEL. This result indicates that Sperm-Halomax assay provides a reliable evaluation of DNA fragmentation of epididymal feline spermatozoa. The DFI seems to be independent from all the measured variables of sperm head morphology and morphometry. Thus, the evaluation of the DNA status of spermatozoa could effectively contribute to the completion of the standard analysis of fresh or frozen semen used in assisted reproductive technologies. Copyright © 2014 Elsevier Inc. All rights reserved.

  6. Marine environmental impact assessment of abalone, Haliotis discus hannai, cage farm in Wan-do, Republic of Korea

    Science.gov (United States)

    Oh, Hyun-Taik; Jung, Rae-Hong; Cho, Yoon-Sik; Hwang, Dong-Woon; Yi, Yong-Min

    2015-12-01

    To assess the marine environmental impacts of abalone, Haliotis discus hannai, cage farms in Wan-do, we monitored the benthic environment on top of the sediment underneath cage farm stations and reference stations. We applied two methods for this assessment. One was the A- and B-investigation of the MOM system (Modeling-On fish farm-Monitoring) developed in Norway. The other was a general environmental monitoring method which is widely used. In this study, we found benthic animals in all samples that belonged to condition 1 which were based on group 1(presence of macrofauna) of the B-investigation method. The values of redox potential (group 2-pH, redox potential) in all samples were above +65 mV belonging to condition 1. Based on sensory results (group 3-gas, color, odor, thickness of deposits), five out of seven experiment samples showed condition 1 while stations 2 and 7 showed condition 2, which have been cultured for 10 years in semi-closed waters. As group 2 takes precedence over group 3, the level of the conditions for B-investigation results consequently showed condition 1 in all stations. We found that pollutants and trace metals in the sediment underneath cage farms were lower than the pollution standard. This led us to conclude that the environmental impacts of the cage farms in this study were not significant.

  7. iTRAQ-Based Identification of Proteins Related to Muscle Growth in the Pacific Abalone, Haliotis discus hannai

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    Jianfang Huang

    2017-10-01

    Full Text Available The abalone Haliotis discus hannai is an important aquaculture species that is grown for human consumption. However, little is known of the genetic mechanisms governing muscle growth in this species, particularly with respect to proteomics. The isobaric tag for relative and absolute quantitation (iTRAQ method allows for sensitive and accurate protein quantification. Our study was the first to use iTRAQ-based quantitative proteomics to investigate muscle growth regulation in H. discus hannai. Among the 1904 proteins identified from six samples, 125 proteins were differentially expressed in large specimens of H. discus hannai as compared to small specimens. In the large specimens, 47 proteins were upregulated and 78 were downregulated. Many of the significant Kyoto Encyclopedia of Genes and Genomes (KEGG pathways, including these differentially expressed proteins, were closely related to muscle growth, including apoptosis, thyroid hormone signaling, regulation of the actin cytoskeleton, and viral myocarditis (p < 0.05. Our quantitative real-time polymerase chain reaction (qRT-PCR analyses suggested that the alterations in expression levels observed in the differentially expressed proteins were consistent with the alterations observed in the encoding mRNAs, indicating the repeatability of our proteomic approach. Our findings contribute to the knowledge of the molecular mechanisms of muscle growth in H. discus hannai.

  8. Effects of abalone (Haliotis discus hannai Ino) gonad polysaccharides on cholecystokinin release in STC-1 cells and its signaling mechanism.

    Science.gov (United States)

    Zhao, Jun; Zhou, Da-Yong; Yang, Jing-Feng; Song, Shuang; Zhang, Ting; Zhu, Ce; Song, Yan-Qing; Yu, Chen-Xu; Zhu, Bei-Wei

    2016-10-20

    Abalone gonad polysaccharide (AGP) -31, -32 and -33 prepared in this study had the molecular weight (MW) of 37.8, 32.2 and 27.5kDa, respectively. They all contained mannose, rhamnose, glucuronic acid, glucose, galactose, xylose, arabinose, and fucose, with very similar monosaccharide profile. All the three polysaccharides could significantly increase the secretion of cholecystokinin (CCK) in STC-1 cells. Among them, AGP-32 showed the strongest effect. However, the low-MW fragments of AGP-32 showed significantly lower activity than AGP-32 itself. It was also found that the inhibitors on calcium-sensing receptor (CaSR), protein kinase A (PKA), Ca(2+)⁄calmodulin-dependent protein kinase (CaMK) II, p38- mitogen-activated protein kinases (MAPK), and an intracellular calcium chelator all inhibited AGP-induced CCK secretion. To conclude, Ca(2+)/calmodulin (CaM)/CaMK, cyclic adenosine monophosphate (cAMP)/PKA and MAPK pathways are all involved in AGP-induced CCK secretion. Copyright © 2016 Elsevier Ltd. All rights reserved.

  9. Proteomic analysis of muscle between hybrid abalone and parental lines Haliotis gigantea Reeve and Haliotis discus hannai Ino.

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    Di, G; Luo, X; You, W; Zhao, J; Kong, X; Ke, C

    2015-06-01

    To understand the potential molecular mechanism of heterosis, protein expression patterns were compared from hybrids of Haliotis gigantea (G) and Haliotis discus hannai (D) using two-dimensional gel electrophoresis (2-DE) and matrix-assisted laser desorption/ionization time-of-flight/time-of-flight analyses. Expression differences were observed in muscle samples from the four groups with 673±21.0 stained spots for H. discus hannai ♀ × H. discus hannai ♂ (DD), 692±25.6 for H. gigantea ♀ × H. gigantea ♂ (GG), 679±16.2 for H. discus hannai ♀ × H. gigantea ♂ (DG) (F1 hybrid) and 700±19 for H. gigantea ♀ × H. discus hannai ♂ (GD) (F1 hybrid). Different 2-DE image muscle protein spots had a mirrored relationship between purebreds and the F1 hybrid, suggesting that all stained spots in F1 hybrid muscle were on 2-DEs from parents. DD and DG clustered together first, and then clustered with GD, whereas the distance of DD and GG was maximal according to hierarchical cluster analysis. We identified 136 differentially expressed protein spots involved in major biological processes, including energy metabolism and stress response. Most energy metabolism proteins were additive, and stress-induced proteins displayed additivity or over-dominance. In these 136 identified protein spots, hybrid offspring with additivity or over-dominance accounted for 68.38%. Data show that a proteomic approach can provide functional prediction of abalone interspecific hybridization.

  10. Molecular identification and functional analysis of two variants of myeloid differentiation factor 88 (MyD88) from disk abalone (Haliotis discus discus).

    Science.gov (United States)

    Priyathilaka, Thanthrige Thiunuwan; Bathige, S D N K; Lee, Seongdo; Lee, Jehee

    2018-02-01

    Myeloid differentiation factor 88 (MyD88) is a crucial adaptor protein of the Toll-like receptor (TLR)- and interleukin 1 receptor-mediated signaling pathways and is involved in a diverse array of inflammatory responses via NF-κB activation. In the present study, two MyD88 variants were identified from disk abalone (Haliotis discus discus) and designated AbMyD88-2 and AbMyD88-X. The deduced AbMyD88-2 and AbMyD88-X comprised 433 and 354 amino acids with predicted molecular masses of 48.85 kDa and 40.17 kDa, respectively. AbMyD88-2 and AbMyD88-X possessed typical MyD88 domain structural features including an N-terminal death domain (DD) and C-terminal toll interleukin 1 receptor (TIR) domain similar to those in mammals. Expression analysis of AbMyD88-2 and AbMyD88-X mRNA at different early embryonic developmental stages of abalone by qPCR revealed that their constitutive expression at all developmental stages analyzed with the considerably higher values at the 16-cell (AbMyD88-2) and morula stages (AbMyD88-X). In unchallenged disk abalones, AbMyD88-2 was highly expressed in muscles, while AbMyD88-X mRNA was predominantly transcribed in hemocytes. Moreover, AbMyD88-2 and AbMyD88-X mRNA were differentially modulated in abalone hemocytes after a challenge with live bacteria (Vibrio parahaemolyticus, Listeria monocytogenes), virus (viral hemorrhagic septicemia virus), and pathogen-associated molecular patterns (lipopolysaccharides and Poly I:C). Overexpression of AbMyD88-2 and AbMyD88-X in HEK293T cells induced the activation of the NF-κB promoter. AbMyD88-2 and AbMyD88-X involvement in inflammatory responses was characterized by their overexpression in RAW264.7 murine macrophage cells. These results revealed comparatively higher NO (Nitric oxide) production, induction of inflammatory mediator genes (iNOS and COX2), and proinflammatory genes (IL1β, IL6 and TNFα) expression in abalone MyD88s-overexpressing cells than in mock control in the presence or absence of LPS

  11. Sperm vacuoles cannot help to differentiate fertile men from infertile men with normal sperm parameter values.

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    Gatimel, N; Léandri, R D; Marino, L; Esquerre-Lamare, C; Parinaud, J

    2014-11-01

    Can the assessment of sperm vacuoles at high magnification contribute to the explanation of idiopathic infertility? The characteristics of sperm head vacuoles (number, area, position) are no different between fertile controls and patients with unexplained infertility. Until now, the assessment of sperm head vacuoles has been focused on a therapeutic goal in the intracytoplasmic morphologically selected sperm injection (IMSI) procedure, but it could be pertinent as a new diagnostic tool for the evaluation of male fertility. This diagnostic test study with blind assessment included a population of 50 fertile men and 51 men with idiopathic infertility. They were selected from September 2011 to May 2013. Fertile men were within couples who had a spontaneous pregnancy in the last 2 years. Infertile men were within couples who had unexplained infertility and were consulting in our centre. After analysis of conventional sperm parameters, we investigated the number, position and area of sperm head vacuoles at high magnification (×6000) with interference contrast using an image analysis software. We also carried out a nuclear status analysis by terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling assay (TUNEL), sperm chromatin structure assay (SCSA) and aniline blue staining. Concerning the vacuoles data, we did not find any significant difference between the two populations. We found no significant correlation between the vacuolar parameters (mean number of vacuoles, relative vacuole area and percentage of spermatozoa with large vacuoles) and either conventional semen parameters, male age or the data from the aniline blue staining, SCSA assay and TUNEL assay. Despite the fact all of the vacuole parameters values were identical in fertile and infertile men, we cannot totally exclude that a very small cause of unexplained infertilities could be related to an excess of sperm vacuoles. In line with its widely debated use as a therapeutic tool, sperm vacuole

  12. Effects of Synthetic Serum Supplementation in Sperm Preparation Media on Sperm Capacitation and Function Test Results

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    Ying-Fu Shih

    2016-01-01

    Full Text Available Albumin supplementation of culture media induces sperm capacitation in assisted reproduction technique cycles. Synthetic serum supplementation is clinically used to replace albumin for preventing transmission of infectious agents. However, the effects of synthetic serum supplementation on sperm capacitation have rarely been investigated. Spermatozoa from 30 men with normal basic semen analysis results were collected, divided into five aliquots, and cultured in capacitating conditions in four combinations of two synthetic serum supplements, serum substitute supplement (SSS and serum protein substitute (SPS, and two fertilization media, Quinns Advantage™ Fertilization (QF and human tubular fluid (HTF media. Reactive oxygen species (ROS levels in spermatozoa were measured through chemiluminescence. Furthermore, acrosome reaction and western blotting for tyrosine phosphorylation were used to evaluate sperm capacitation. HTF+SSS had significantly higher ROS levels than QF+SPS did (11,725 ± 1,172 versus 6,278 ± 864 relative light units. In addition, the spermatozoa cultured in QF+SPS had lower motility, acrosome reaction rates, and tyrosine phosphorylation levels compared with those cultured in HTF+SSS. In conclusion, the effects of synthetic serum supplementation on sperm capacitation varied according to the combination of media. These differences may lead to variations in spermatozoon ROS levels, thus affecting sperm function test results.

  13. Sperm Impairment by Sperm Agglutinating Factor Isolated from Escherichia coli: Receptor Specific Interactions

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    Kiranjeet Kaur

    2013-01-01

    Full Text Available In an earlier work done in our laboratory, we have been able to isolate a sperm agglutinating strain of Escherichia coli from the semen sample of a male attending infertility clinic. Further, factor responsible for sperm agglutination (SAF was isolated and purified, and, using SAF as a tool, corresponding SAF binding receptor from human spermatozoa has been purified. Characterization of SAF and SAF binding receptor using MALDI-TOF showed homology to glutamate decarboxylase and MHC class I molecule, respectively. Coincubation of SAF with spermatozoa not only resulted in spermagglutination but could also compromise other sperm parameters, namely, Mg2+ dependent ATPase activity and apoptosis. Intravaginal administration of SAF could lead to infertility in Balb/c mice. SAF induced impairment of sperm parameters, and infertility was observed to be due to interaction of SAF with sperm surface receptor component as, when purified receptor was introduced, receptor completely inhibited all the detrimental effects induced by SAF. From these results, it could be concluded that interaction of SAF with spermatozoa is receptor mediated.

  14. Evaluation of Lasting Effects of Heat Stress on Sperm Profile and Oxidative Status of Ram Semen and Epididymal Sperm

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    Thais Rose dos Santos Hamilton

    2016-01-01

    Full Text Available Higher temperatures lead to an increase of testicular metabolism that results in spermatic damage. Oxidative stress is the main factor responsible for testicular damage caused by heat stress. The aim of this study was to evaluate lasting effects of heat stress on ejaculated sperm and immediate or long-term effects of heat stress on epididymal sperm. We observed decrease in motility and mass motility of ejaculated sperm, as well as an increase in the percentages of sperm showing major and minor defects, damaged plasma and acrosome membranes, and a decrease in the percentage of sperm with high mitochondrial membrane potential in the treated group until one spermatic cycle. An increased enzymatic activity of glutathione peroxidase and an increase of stressed cells were observed in ejaculated sperm of the treated group. A decrease in the percentage of epididymal sperm with high mitochondrial membrane potential was observed in the treated group. However, when comparing immediate and long-term effects, we observed an increase in the percentage of sperm with low mitochondrial membrane potential. In conclusion, testicular heat stress induced oxidative stress that led to rescuable alterations after one spermatic cycle in ejaculated sperm and also after 30 days in epididymal sperm.

  15. Matrine Inhibits Mouse Sperm Function by Reducing Sperm [Ca2+]i and Phospho-ERK1/2

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    Tao Luo

    2015-01-01

    Full Text Available Background: Matrine is a bioactive alkaloid that has a variety of pharmacological effects and is widely used in Chinese medicine. However, its effects on male reproduction are not well known. In this study, we aimed to investigate the in vitro toxicity of matrine on mature mouse sperm. Methods: Mouse cauda epididymal sperm were exposed to matrine (10-200 µM in vitro. The viability, motility, capacitation, acrosome reaction and fertilization ability of the mouse sperm were examined. Furthermore, the intracellular calcium concentration ([Ca2+]i, calcium (Catsper and potassium (Ksper currents, and phosphorylation of extracellular signal regulated kinases 1/2 (p-ERK1/2 of the sperm were analyzed. Results: After exposure to 100 µM or more of matrine, mouse cauda epididymal sperm exhibited a significant reduction in total motility, progressive motility, linear velocity and acrosome reaction rate induced by Ca2+ ionophore A23187. As a result, the fertilization ability of mouse sperm was remarkably decreased by matrine. Our data further demonstrated that matrine significantly reduced sperm [Ca2+]i and [Ca2+]i-related p-ERK1/2; however, both the CatSper and KSper currents, which are thought to interactively regulate Ca2+ influx in sperm, were not affected by matrine. Conclusion: Our findings indicate that matrine inhibits mouse sperm function by reducing sperm [Ca2+]i and suppressing the phosphorylation of ERK1/2.

  16. Presence and function of dopamine transporter (DAT in stallion sperm: dopamine modulates sperm motility and acrosomal integrity.

    Directory of Open Access Journals (Sweden)

    Javier A Urra

    Full Text Available Dopamine is a catecholamine with multiple physiological functions, playing a key role in nervous system; however its participation in reproductive processes and sperm physiology is controversial. High dopamine concentrations have been reported in different portions of the feminine and masculine reproductive tract, although the role fulfilled by this catecholamine in reproductive physiology is as yet unknown. We have previously shown that dopamine type 2 receptor is functional in boar sperm, suggesting that dopamine acts as a physiological modulator of sperm viability, capacitation and motility. In the present study, using immunodetection methods, we revealed the presence of several proteins important for the dopamine uptake and signalling in mammalian sperm, specifically monoamine transporters as dopamine (DAT, serotonin (SERT and norepinephrine (NET transporters in equine sperm. We also demonstrated for the first time in equine sperm a functional dopamine transporter using 4-[4-(Dimethylaminostyryl]-N-methylpyridinium iodide (ASP(+, as substrate. In addition, we also showed that dopamine (1 mM treatment in vitro, does not affect sperm viability but decreases total and progressive sperm motility. This effect is reversed by blocking the dopamine transporter with the selective inhibitor vanoxerine (GBR12909 and non-selective inhibitors of dopamine reuptake such as nomifensine and bupropion. The effect of dopamine in sperm physiology was evaluated and we demonstrated that acrosome integrity and thyrosine phosphorylation in equine sperm is significantly reduced at high concentrations of this catecholamine. In summary, our results revealed the presence of monoamine transporter DAT, NET and SERT in equine sperm, and that the dopamine uptake by DAT can regulate sperm function, specifically acrosomal integrity and sperm motility.

  17. Proteins associated with critical sperm functions and sperm head shape are differentially expressed in morphologically abnormal bovine sperm induced by scrotal insulation

    NARCIS (Netherlands)

    Saadi, H.A.S.; Riemsdijk, van E.L.C.; Dance, A.L.; Rajamanickam, G.D.; Kastelic, J.P.; Thundathil, J.C.

    2013-01-01

    The objective was to investigate expression patterns of proteins in pyriform sperm, a common morphological abnormality in bull sperm. Ejaculates were collected from sexually mature Holstein bulls (n = 3) twice weekly for 10 weeks (pre-thermal insult samples). Testicular temperature was elevated in

  18. Mitogen-activated protein kinase 1 from disk abalone (Haliotis discus discus): Roles in early development and immunity-related transcriptional responses.

    Science.gov (United States)

    Perera, N C N; Godahewa, G I; Lee, Jehee

    2016-12-01

    Mitogen-activated protein kinase (MAPK) is involved in the regulation of cellular events by mediating signal transduction pathways. MAPK1 is a member of the extracellular-signal regulated kinases (ERKs), playing roles in cell proliferation, differentiation, and development. This is mainly in response to growth factors, mitogens, and many environmental stresses. In the current study, we have characterized the structural features of a homolog of MAPK1 from disk abalone (AbMAPK1). Further, we have unraveled its expressional kinetics against different experimental pathogenic infections or related chemical stimulants. AbMAPK1 harbors a 5' untranslated region (UTR) of 23 bps, a coding sequence of 1104 bps, and a 3' UTR of 448 bp. The putative peptide comprises a predicted molecular mass of 42.2 kDa, with a theoretical pI of 6.28. Based on the in silico analysis, AbMAPK1 possesses two N-glycosylation sites, one S_TK catalytic domain, and a conserved His-Arg-Asp domain (HRD). In addition, a conservative glycine rich ATP-phosphate-binding loop and a threonine-x-tyrosine motif (TEY) important for the autophosphorylation were also identified in the protein. Homology assessment of AbMAPK1 showed several conserved regions, and ark clam (Aplysia californica) showed the highest sequence identity (87.9%). The phylogenetic analysis supported close evolutionary kinship with molluscan orthologs. Constitutive expression of AbMAPK1 was observed in six different tissues of disk abalone, with the highest expression in the digestive tract, followed by the gills and hemocytes. Highest AbMAPK1 mRNA expression level was detected at the trochophore developmental stage, suggesting its role in abalone cell differentiation and proliferation. Significant modulation of AbMAPK1 expression under pathogenic stress suggested its putative involvement in the immune defense mechanism. Copyright © 2016 Elsevier Ltd. All rights reserved.

  19. Molluscan death effector domain (DED)-containing caspase-8 gene from disk abalone (Haliotis discus discus): molecular characterization and expression analysis.

    Science.gov (United States)

    Lee, Youngdeuk; De Zoysa, Mahanama; Whang, Ilson; Lee, Sukkyoung; Kim, Yucheol; Oh, Chulhong; Choi, Cheol Young; Yeo, Sang-Yeob; Lee, Jehee

    2011-02-01

    The caspase family represents aspartate-specific cysteine proteases that play key roles in apoptosis and immune signaling. In this study, we cloned the first death effector domain (DED)-containing molluscan caspase-8 gene from disk abalone (Haliotis discus discus), which is named as hdCaspase-8. The full-length hdCaspase was 2855 bp, with a 1908 bp open reading frame encoding 636 amino acids. The hdCaspase-8 had 72 kDa predicted molecular mass with an estimated isoelectric point (PI) of 6.0. The hdCaspase-8 amino acid sequence contained the characteristic feature of an N-terminal two DED, a C-terminal catalytic domain and the caspase family cysteine active site ⁵¹³KPKLFFLQACQG⁵²⁴. Phylogenetic analysis results showed that hdCaspase-8 is more similar to the invertebrate Tubifex tubifex (sludge worm) caspase-8. Real-time RT-PCR results showed that hdCaspase-8 constitutively and ubiquitously expressed in all tested tissue of unchallenged disk abalone. The basal expression level of hdCaspase-8 in gill tissue was higher than all other tested tissues. The hdCaspase-8 mRNA expression in gill and hemocytes was significantly up-regulated by exposure to bacteria (Vibrio alginolyticus, Vibrio parahemolyticus and Listeria monocytogenes) and VHSV (viral hemorrhagic septicemia virus), as compared to control animals. These results suggest that hdCaspase-8 may be involved in immune response reactions in disk abalone. Copyright © 2010 Elsevier Ltd. All rights reserved.

  20. Metabolic engineering for L-lysine production by Corynebacterium glutamicum.

    Science.gov (United States)

    de Graaf, A A; Eggeling, L; Sahm, H

    2001-01-01

    Corynebacterium glutamicum has been used since several decades for the large-scale production of amino acids, esp. L-glutamate and L-lysine. After initial successes of random mutagenesis and screening approaches, further strain improvements now require a much more rational design, i.e. metabolic engineering. Not only recombinant DNA technology but also mathematical modelling of metabolism as well as metabolic flux analysis represent important metabolic engineering tools. This review covers as state-of-the-art examples of these techniques the genetic engineering of the L-lysine biosynthetic pathway resulting in a vectorless strain with significantly increased dihydrodipicolinate synthase activity, and the detailed metabolic flux analysis by 13C isotopomer labelling strategies of the anaplerotic enzyme activities in C. glutamicum resulting in the identification of gluconeogenic phosphoenolpyruvate carboxykinase as a limiting enzyme.

  1. MOLECULAR DYNAMICS SIMULATION OF LYSINE DENDRIMER AND SEMAX PEPTIDES INTERACTION

    Directory of Open Access Journals (Sweden)

    E. V. Popova

    2016-07-01

    Full Text Available The paper deals with the possibility of complex formation of therapeutic Semax peptides with lysine dendrimer by molecular modeling methods. Dendrimers are often used for delivery of drugs and biological molecules (e.g., DNA, peptides and polysaccharides. Since lysine dendrimers are less toxic than conventional synthetic dendrimers (e.g., polyamidoamine (PAMAM dendrimer, we chose them and studied two systems containing dendrimer and the different number of Semax peptides. The study was carried out by molecular dynamics method. It was obtained that the stable complexes were formed in both cases. The equilibrium structures of these complexes were investigated. These complexes can be used in the future in therapy of various diseases as Semax peptides have significant antioxidant, antihypoxic and neuroprotecting action.

  2. Method-related estimates of sperm vitality.

    Science.gov (United States)

    Cooper, Trevor G; Hellenkemper, Barbara

    2009-01-01

    Comparison of methods that estimate viability of human spermatozoa by monitoring head membrane permeability revealed that wet preparations (whether using positive or negative phase-contrast microscopy) generated significantly higher percentages of nonviable cells than did air-dried eosin-nigrosin smears. Only with the latter method did the sum of motile (presumed live) and stained (presumed dead) preparations never exceed 100%, making this the method of choice for sperm viability estimates.

  3. Methods for cryopreservation of guinea fowl sperm.

    Directory of Open Access Journals (Sweden)

    Éva Váradi

    Full Text Available Conservation of indigenous poultry species is an important part of the new Hungarian agricultural strategy. Semen cryopreservation is the most practical method for the long term storage of poultry genetic material. The objective was to compare four protocols for cryopreservation of guinea fowl sperm (slow and fast programmable, freezing in nitrogen vapor, and pellet and three cryoprotectants (10% ethylene glycol, 6% dimethyl-formamide and 6% dimethyl-acetamide. The efficiency of the methods was examined by in vitro tests (subjective motility scoring, sperm concentration, morphological and live/dead sperm analysis with eosin-aniline staining. Thereafter, the two most promising methods were tested by artificial insemination of frozen-thawed semen (3 times a week for 3 weeks using 300 million spermatozoa/hen, followed by candling of incubated eggs, assessment of fertilization, embryonic death, and hatching rate. The survival rate of live, intact spermatozoa was greatest (p≤0.05 in pellet method and the slow programmable protocol (with 10% ethylene glycol (28.6 and 23.5%. The two best protocols (based on in vitro assessment of post-thaw semen quality were subsequently tested in vivo with artificial insemination. The pellet method yielded a 64% fertility rate compared to slow protocol with only 30% fertility. Regardless, both freezing protocols significantly increased embryonic deaths compared to the control group (16,7; 9,1 and 8,3%, respectively. During the 3-week in vivo trial, fertility increased and early embryonic death decreased over time. According to the results the guinea fowl sperm could tolerate the fast freezing in pellet better than the slower freezing rates and resulted acceptable fertility rate.

  4. Formation and development of sperms in angiosperms

    Directory of Open Access Journals (Sweden)

    S. Tatintseva

    2014-01-01

    Full Text Available Spermiogelnesis has been studied in a large number of Angiosperm species characterizing different levels of phylogenetic system. The formation and development of male gamets can be described as formation of sperm cells that undergo ontogenesis which stimulates changes up to full maturity of the pollen grain. The process of ontogenesis is aimed at creating a suitable delivery system by the pollen tube to a female gamete.

  5. Sperm navigation along helical paths in 3D chemoattractant landscapes

    Science.gov (United States)

    Jikeli, Jan F.; Alvarez, Luis; Friedrich, Benjamin M.; Wilson, Laurence G.; Pascal, René; Colin, Remy; Pichlo, Magdalena; Rennhack, Andreas; Brenker, Christoph; Kaupp, U. Benjamin

    2015-08-01

    Sperm require a sense of direction to locate the egg for fertilization. They follow gradients of chemical and physical cues provided by the egg or the oviduct. However, the principles underlying three-dimensional (3D) navigation in chemical landscapes are unknown. Here using holographic microscopy and optochemical techniques, we track sea urchin sperm navigating in 3D chemoattractant gradients. Sperm sense gradients on two timescales, which produces two different steering responses. A periodic component, resulting from the helical swimming, gradually aligns the helix towards the gradient. When incremental path corrections fail and sperm get off course, a sharp turning manoeuvre puts sperm back on track. Turning results from an `off' Ca2+ response signifying a chemoattractant stimulation decrease and, thereby, a drop in cyclic GMP concentration and membrane voltage. These findings highlight the computational sophistication by which sperm sample gradients for deterministic klinotaxis. We provide a conceptual and technical framework for studying microswimmers in 3D chemical landscapes.

  6. Aspermy, sperm quality and radiation in Chernobyl birds.

    Science.gov (United States)

    Møller, Anders Pape; Bonisoli-Alquati, Andrea; Mousseau, Timothy A; Rudolfsen, Geir

    2014-01-01

    Following the Chernobyl nuclear power plant accident, large amounts of radionuclides were emitted and spread in the environment. Animals living in such contaminated areas are predicted to suffer fitness costs including reductions in the quality and quantity of gametes. We studied whether aspermy and sperm quality were affected by radioactive contamination by examining ejaculates from wild caught birds breeding in areas varying in background radiation level by more than three orders of magnitude around Chernobyl, Ukraine. The frequency of males with aspermy increased logarithmically with radiation level. While 18.4% of males from contaminated areas had no sperm that was only the case for 3.0% of males from uncontaminated control areas. Furthermore, there were negative relationships between sperm quality as reflected by reduced sperm velocity and motility, respectively, and radiation. Our results suggest that radioactive contamination around Chernobyl affects sperm production and quality. We are the first to report an interspecific difference in sperm quality in relation to radioactive contamination.

  7. Progress and challenges of fish sperm vitrification: A mini review.

    Science.gov (United States)

    Xin, Miaomiao; Siddique, Mohammad Abdul Momin; Dzyuba, Borys; Cuevas-Uribe, Rafael; Shaliutina-Kolešová, Anna; Linhart, Otomar

    2017-08-01

    To survive low temperature is required for a long-term storage (cryopreservation), cells should be vitrified to a state in which intracellular water is solidified without ice crystal formation. Two different approaches are described for fish sperm cryopreservation: 1) sperm conventional cryopreservation, in which extracellular water is partially crystallized and 2) sperm vitrification, in which both intra- and extra-cellular liquids are vitrified. Sperm vitrification has been applied to some fish species with limited success. Traditional vitrification requires rapid cooling/warming rates, small sample carriers, and using high permeable cryoprotectant concentrations. The latter cause cytotoxic effects which must be well managed and will require continuous effort to match an appropriate cryoprotectant with suitable apparatus and warming methods. Novel cryoprotectant-free sperm vitrification approach has been applied to several fishes. This review summarizes development of basic procedures and discusses advantages and disadvantages of vitrification when applied it to fish sperm. Copyright © 2017 Elsevier Inc. All rights reserved.

  8. Mating duration and sperm precedence in the spider Linyphia triangularis

    DEFF Research Database (Denmark)

    Weldingh, Ditte L.; Toft, Søren; Larsen, Ole Næsbye

    2011-01-01

    , especially the males, are able to influence the outcome of mating for their own benefit. We studied the linyphiid spider Linyphia triangularis in which mating follows a strict sequence during which the male inducts two droplets of sperm and transfers them to the female. We performed sperm competition...... experiments (sterile-male technique) including four treatments, in which the copulation of the first male was interrupted at prescribed phases of the mating sequence, while the second male was allowed a complete mating. Second males spent a shorter time than first males on the behaviours prior to sperm...... transfer, but the amount of sperm (2 droplets) and the time spent in sperm transfer were independent of the females’ mating status. The proportion of females accepting the second male depended on the mating duration of the first male, i.e. whether the first male had transferred one or two sperm droplets...

  9. Association between chronic pain and the sperm motion characteristics

    DEFF Research Database (Denmark)

    Dardmeh, Fereshteh; Alipour, Hiva; Nielsen, Hans Ingolf

    2015-01-01

    Sex hormones play an important role in pain in many chronic pain conditions. Relationship between chronic pain and sperm quality has not been investigated thoroughly and may provide an insight to better understanding, management and treatment of cases where chronic pain and male sub-fertility co......-exist. Neat (fresh semen) and processed sperm from 15 males with orthopedic chronic pain (CP) were assessed and compared with 15 healthy age matched controls. Sperm analysis was performed using the SCA computer-aided sperm analyzer. There was no significant difference in any parameters of the neat semen...... significantly higher in the processed sample of the CP group. This study demonstrated that chronic pain does not affect the sperm morphology, total concentration and motility based on conventional analysis but has significant influence at the level of sperm motion kinetics which could prove to be clinically...

  10. Accountings of Selecting Sperm on the (ethical) border

    DEFF Research Database (Denmark)

    Willum Adrian, Stine

    During the past years, Denmark has become a destination for fertility travellers, in need for donated sperm. Today, treatment is possible no matter of marital status or sexuality. Furthermore, users of sperm donation can choose between anonymous and non-anonymous sperm, with either basic...... will analyze how women and their partners crossing the Danish border accounts for their donor choice, and their practices of choosing. I will be exploring the question: How are women and their partners accounting for choosing a sperm donor, crossing the (ethical) border to Denmark? The use of the term......-sited ethnographic fieldwork at a Danish private fertility clinic, and a Danish sperm bank. The presentation will mainly draw on interviews with fertility travellers going through treatment with donor sperm in Denmark....

  11. Some Reflections on Intracytoplasmic Morphologically Selected Sperm Injection

    Directory of Open Access Journals (Sweden)

    Thomas Ebner

    2014-07-01

    Full Text Available Although intracytoplasmic sperm injection (ICSI allows proper fertilization in most cases of male sub fertility, it is one of the most unphysiological techniques in assisted reproductive technologies (ART. Thus, over the last decade, researchers have tried to improve sperm observation with higher-resolution microscopy techniques such as the intracytoplasmic morphologically selected sperm injection (IMSI technique. In order to identify literatures for this review, the PubMed database was searched from 2000 onwards using the terms IMSI, motile sperm organelle morphology examination (MSOME and sperm vacuole. Approximately 10 years after the introduction of the MSOME and IMSI procedures, several questions related to the prevalence, origin, location, and clinical consequences of sperm vacuoles have not yet been clarified. It seems that IMSI as a routine application is not state of the art and the only confirmed indications for IMSI are recurrent implantation failure following ICSI and severe male factor.

  12. Acetylation site specificities of lysine deacetylase inhibitors in human cells

    DEFF Research Database (Denmark)

    Schölz, Christian; Weinert, Brian Tate; Wagner, Sebastian A

    2015-01-01

    Lysine deacetylases inhibitors (KDACIs) are used in basic research, and many are being investigated in clinical trials for treatment of cancer and other diseases. However, their specificities in cells are incompletely characterized. Here we used quantitative mass spectrometry (MS) to obtain acety......1-α, providing a possible mechanistic explanation of its adverse, pro-inflammatory effects. Our results offer a systems view of KDACI specificities, providing a framework for studying function of acetylation and deacetylases....

  13. PLMD: An updated data resource of protein lysine modifications.

    Science.gov (United States)

    Xu, Haodong; Zhou, Jiaqi; Lin, Shaofeng; Deng, Wankun; Zhang, Ying; Xue, Yu

    2017-05-20

    Post-translational modifications (PTMs) occurring at protein lysine residues, or protein lysine modifications (PLMs), play critical roles in regulating biological processes. Due to the explosive expansion of the amount of PLM substrates and the discovery of novel PLM types, here we greatly updated our previous studies, and presented a much more integrative resource of protein lysine modification database (PLMD). In PLMD, we totally collected and integrated 284,780 modification events in 53,501 proteins across 176 eukaryotes and prokaryotes for up to 20 types of PLMs, including ubiquitination, acetylation, sumoylation, methylation, succinylation, malonylation, glutarylation, glycation, formylation, hydroxylation, butyrylation, propionylation, crotonylation, pupylation, neddylation, 2-hydroxyisobutyrylation, phosphoglycerylation, carboxylation, lipoylation and biotinylation. Using the data set, a motif-based analysis was performed for each PLM type, and the results demonstrated that different PLM types preferentially recognize distinct sequence motifs for the modifications. Moreover, various PLMs synergistically orchestrate specific cellular biological processes by mutual crosstalks with each other, and we totally found 65,297 PLM events involved in 90 types of PLM co-occurrences on the same lysine residues. Finally, various options were provided for accessing the data, while original references and other annotations were also present for each PLM substrate. Taken together, we anticipated the PLMD database can serve as a useful resource for further researches of PLMs. PLMD 3.0 was implemented in PHP + MySQL and freely available at http://plmd.biocuckoo.org. Copyright © 2017 Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, and Genetics Society of China. Published by Elsevier Ltd. All rights reserved.

  14. Molecular identification and functional delineation of a glutathione reductase homolog from disk abalone (Haliotis discus discus): Insights as a potent player in host antioxidant defense.

    Science.gov (United States)

    Herath, H M L P B; Wickramasinghe, P D S U; Bathige, S D N K; Jayasooriya, R G P T; Kim, Gi-Young; Park, Myoung Ae; Kim, Chul; Lee, Jehee

    2017-01-01

    Glutathione reductase (GSR) is an enzyme that catalyzes the biochemical conversion of oxidized glutathione (GSSG) into the reduced form (GSH). Since the ratio between the two forms of glutathione (GSH/GSSG) is important for the optimal function of GSH to act as an antioxidant against H 2 O 2 , the contribution of GSR as an enzymatic regulatory agent to maintain the proper ratio is essential. Abalones are marine mollusks that frequently encounter environmental factors that can trigger the overproduction of reactive oxygen species (ROS) such as H 2 O 2 . Therefore, we conducted the current study to reveal the molecular and functional properties of a GSR homolog in the disk abalone, Haliotis discus discus. The identified cDNA sequence (2325 bp) has a 1356 bp long open reading frame (ORF), coding for a 909 bp long amino acid sequence, which harbors a pyridine nucleotide-disulfide oxidoreductase domain (171-246 aa), a pyridine nucleotide-disulfide oxidoreductase dimerization domain, and a NAD(P)(+)-binding Rossmann fold superfamily signature domain. Four functional residues: the FAD binding site, glutathione binding site, NADPH binding motif, and assembly domain were identified to be conserved among the other species. The recombinant abalone GSR (rAbGSR) exhibited detectable activity in a standard glutathione reductase activity assay. The optimum pH and optimal temperature for the reaction were found to be 7.0 and 50 °C, respectively, while the ionic strength of the medium had no effect. The enzymatic reaction was vastly inhibited by Cu +2 and Cd +2 ions. A considerable effect of cellular protection was detected with a disk diffusion assay conducted with rAbGSR. Moreover, an MTT assay and flow cytometry confirmed the significance of the protective role of rAbGSR in cell function. Furthermore, AbGSR was found to be ubiquitously distributed in different types of abalone tissues. AbGSR mRNA expression was significantly upregulated in response to three immune challenges

  15. Molecular characterization of kappa class glutathione S-transferase from the disk abalone (Haliotis discus discus) and changes in expression following immune and stress challenges.

    Science.gov (United States)

    Sandamalika, W M Gayashani; Priyathilaka, Thanthrige Thiunuwan; Liyanage, D S; Lee, Sukkyoung; Lim, Han-Kyu; Lee, Jehee

    2018-04-02

    Glutathione S-transferase (GST; EC 2.5.1.18) isoenzymes represent a complex group of proteins that are involved in phase II detoxification in several organisms. In this study, GST kappa (GSTκ) from the disk abalone (Haliotis discus discus; AbGSTκ) was characterized at both the transcriptional and functional levels to determine its potential capacity to perform as a detoxification agent under conditions of different stress. The predicted AbGSTκ protein consists of 227 amino acids, with a predicted molecular weight of 25.6 kDa and a theoretical isoelectric point (pI) of 7.78. In silico analysis reveals that AbGSTκ is a disulfide bond formation protein A (DsbA), consisting of a thioredoxin domain, GSH binding sites (G-sites), and a catalytic residue. In contrast, no hydrophobic ligand binding site (H-site), or signal peptides, were detected. AbGSTκ showed the highest sequence identity with the orthologue from pufferfish (Takifugu obscurus) (60.0%). In a phylogenetic tree, AbGSTκ clustered closely together with other fish GSTκs, and was evolutionarily distanced from other cytosolic GSTs. The predicted three-dimensional structure clearly demonstrates that the dimer adopts a butterfly-like shape. A tissue distribution analysis revealed that GSTκ was highly expressed in the digestive tract, suggesting it has detoxification ability. Depending on the tissue and time, AbGSTκ showed different expression patterns, and levels of expression, following challenge of the abalone with immune stimulants. Enzyme kinetics of the purified recombinant proteins demonstrated its conjugating ability using 1-Chloro-2,4-dinitrobenzene (CDNB) and glutathione (GSH) as substrates, and suggested it has a low affinity for both substrates. The optimum temperature and pH for the rAbGSTκ GSH: CDNB conjugating activity were found to be 35 °C and pH 8, respectively indicating that the abalone is well adapted to a wide range of environmental conditions. Cibacron blue (100 μM) was

  16. Puma (Puma concolor) epididymal sperm morphometry.

    Science.gov (United States)

    Cucho, Hernán; Alarcón, Virgilio; Ordóñez, César; Ampuero, Enrique; Meza, Aydee; Soler, Carles

    2016-01-01

    The Andean puma (Puma concolor) has not been widely studied, particularly in reference to its semen characteristics. The aim of the present study was to define the morphometry of puma sperm heads and classify their subpopulations by cluster analysis. Samples were recovered postmortem from two epididymides from one animal and prepared for morphological observation after staining with the Hemacolor kit. Morphometric data were obtained from 581 spermatozoa using a CASA-Morph system, rendering 13 morphometric parameters. The principal component (PC) analysis was performed followed by cluster analysis for the establishment of subpopulations. Two PC components were obtained, the first related to size and the second to shape. Three subpopulations were observed, corresponding to elongated and intermediate-size sperm heads and acrosomes, to large heads with large acrosomes, and to small heads with short acrosomes. In conclusion, puma spermatozoa showed no uniform sperm morphology but three clear subpopulations. These results should be used for future work in the establishment of an adequate germplasm bank of this species.

  17. Hyperbranched lysine-arginine copolymer for gene delivery.

    Science.gov (United States)

    Peng, Qi; Zhu, Jianjun; Yu, Yongsheng; Hoffman, Lee; Yang, Xingkun

    2015-01-01

    Based on the reactivity of amine groups and carboxyl groups of L-lysine and L-arginine, thermal polymerization of these two natural amino acids results in hyperbranched lysine-arginine copolymers (P-lys-argX, where X refers to the relevant molar ratio of arginine to lysine). Hyperbranched polylysine (P-lys) and two derivatives (P-lys-arg0.10 and P-lys-arg0.20) have been prepared. The arginine-rich hyperbranched polymers can interact with plasmid DNA to form nano-sized particles. The polyplexes were physicochemically analyzed by agarose gel electrophoresis, dynamic light scattering, and zeta potential measurements. Furthermore, their transfection efficiency was assessed, employing COS-7, 293T, and HeLa cell lines. It was found that P-lys showed poorly in its ability of condensation with DNA and transfection efficiency. On the other hand, arginine-rich products resulted to significant enhancement of its transfection efficiency, which is dependent on the content of arginine in the polymers, and the cell line used. P-lys-arg0.20 exhibited better transfection efficiency under all the condition studied. Besides, P-lys-arg0.20 showed lower toxicity in COS-7 cells.

  18. Chemical mechanisms of histone lysine and arginine modifications.

    Science.gov (United States)

    Smith, Brian C; Denu, John M

    2009-01-01

    Histone lysine and arginine residues are subject to a wide array of post-translational modifications including methylation, citrullination, acetylation, ubiquitination, and sumoylation. The combinatorial action of these modifications regulates critical DNA processes including replication, repair, and transcription. In addition, enzymes that modify histone lysine and arginine residues have been correlated with a variety of human diseases including arthritis, cancer, heart disease, diabetes, and neurodegenerative disorders. Thus, it is important to fully understand the detailed kinetic and chemical mechanisms of these enzymes. Here, we review recent progress towards determining the mechanisms of histone lysine and arginine modifying enzymes. In particular, the mechanisms of S-adenosyl-methionine (AdoMet) dependent methyltransferases, FAD-dependent demethylases, iron dependent demethylases, acetyl-CoA dependent acetyltransferases, zinc dependent deacetylases, NAD(+) dependent deacetylases, and protein arginine deiminases are covered. Particular attention is paid to the conserved active-site residues necessary for catalysis and the individual chemical steps along the catalytic pathway. When appropriate, areas requiring further work are discussed.

  19. Treating boar sperm with cholesterol-loaded cyclodextrins or cyclodextrins prior to cryopreservation: effects on post-thaw in vitro sperm quality of sperm cryopreserved in different freezing extenders.

    OpenAIRE

    BLANCH TORRES, EVA

    2016-01-01

    [EN] Cryopreserved boar sperm is not used extensively for artificial insemination due to poor fertility rates of the sperm after freezing and thawing. The sperm membrane is damaged when cooled from body temperature to 5 ºC (cold shock), as well as during the freeze-thaw process. Increasing the cholesterol content of boar sperm membranes could increase their post-thaw survival, similarly to other species that are cold shock sensitive. Cholesterol can be easily added to sperm membranes using ch...

  20. A dictionary learning approach for human sperm heads classification.

    Science.gov (United States)

    Shaker, Fariba; Monadjemi, S Amirhassan; Alirezaie, Javad; Naghsh-Nilchi, Ahmad Reza

    2017-12-01

    To diagnose infertility in men, semen analysis is conducted in which sperm morphology is one of the factors that are evaluated. Since manual assessment of sperm morphology is time-consuming and subjective, automatic classification methods are being developed. Automatic classification of sperm heads is a complicated task due to the intra-class differences and inter-class similarities of class objects. In this research, a Dictionary Learning (DL) technique is utilized to construct a dictionary of sperm head shapes. This dictionary is used to classify the sperm heads into four different classes. Square patches are extracted from the sperm head images. Columnized patches from each class of sperm are used to learn class-specific dictionaries. The patches from a test image are reconstructed using each class-specific dictionary and the overall reconstruction error for each class is used to select the best matching class. Average accuracy, precision, recall, and F-score are used to evaluate the classification method. The method is evaluated using two publicly available datasets of human sperm head shapes. The proposed DL based method achieved an average accuracy of 92.2% on the HuSHeM dataset, and an average recall of 62% on the SCIAN-MorphoSpermGS dataset. The results show a significant improvement compared to a previously published shape-feature-based method. We have achieved high-performance results. In addition, our proposed approach offers a more balanced classifier in which all four classes are recognized with high precision and recall. In this paper, we use a Dictionary Learning approach in classifying human sperm heads. It is shown that the Dictionary Learning method is far more effective in classifying human sperm heads than classifiers using shape-based features. Also, a dataset of human sperm head shapes is introduced to facilitate future research. Copyright © 2017 Elsevier Ltd. All rights reserved.

  1. Parallel Evolution of Sperm Hyper-Activation Ca2+ Channels.

    Science.gov (United States)

    Cooper, Jacob C; Phadnis, Nitin

    2017-07-01

    Sperm hyper-activation is a dramatic change in sperm behavior where mature sperm burst into a final sprint in the race to the egg. The mechanism of sperm hyper-activation in many metazoans, including humans, consists of a jolt of Ca2+ into the sperm flagellum via CatSper ion channels. Surprisingly, all nine CatSper genes have been independently lost in several animal lineages. In Drosophila, sperm hyper-activation is performed through the cooption of the polycystic kidney disease 2 (pkd2) Ca2+ channel. The parallels between CatSpers in primates and pkd2 in Drosophila provide a unique opportunity to examine the molecular evolution of the sperm hyper-activation machinery in two independent, nonhomologous calcium channels separated by > 500 million years of divergence. Here, we use a comprehensive phylogenomic approach to investigate the selective pressures on these sperm hyper-activation channels. First, we find that the entire CatSper complex evolves rapidly under recurrent positive selection in primates. Second, we find that pkd2 has parallel patterns of adaptive evolution in Drosophila. Third, we show that this adaptive evolution of pkd2 is driven by its role in sperm hyper-activation. These patterns of selection suggest that the evolution of the sperm hyper-activation machinery is driven by sexual conflict with antagonistic ligands that modulate channel activity. Together, our results add sperm hyper-activation channels to the class of fast evolving reproductive proteins and provide insights into the mechanisms used by the sexes to manipulate sperm behavior. © The Author 2017. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.

  2. Good Quality - the Routinization of Sperm Banking in China

    DEFF Research Database (Denmark)

    Wahlberg, Ayo

    Good Quality is an assemblage ethnography of how sperm banking came to be routinized in China. Based on 8 years of episodic fieldwork at China’s oldest and largest sperm bank in Changsha, Hunan province, the book meticulously chronicles how, beginning in the early 1980s, a unique style of sperm...... banking would emerge in China, shaped by the cultural, juridical, economic and social configurations that make up China’s restrictive reproductive complex....

  3. A sterile sperm caste protects brother fertile sperm from female-mediated death in Drosophila pseudoobscura

    DEFF Research Database (Denmark)

    Holman, Luke; Snook, Rhonda R

    2008-01-01

    Spermicide (i.e., female-mediated sperm death) is an understudied but potentially widespread phenomenon that has important ramifications for the study of sexual conflict, postcopulatory sexual selection, and fertility [1, 2]. Males are predicted to evolve adaptations against spermicide, but few...... antispermicidal mechanisms have been definitively identified. One such adaptation may be the enigmatic infertile sperm morphs or "parasperm" produced by many species, which have been hypothesized to protect their fertile brother "eusperm" from spermicide [2, 3]. Here, we show that female Drosophila pseudoobscura...... reproductive tracts are spermicidal and that the survival of eusperm after exposure to the female tract is highest when males produce many parasperm. This study clarifies the adaptive significance of infertile sperm castes, which has remained elusive in Drosophila and other taxa despite much recent interest [2...

  4. On the relative effect of spawning asynchrony, sperm quantity and sperm quality on paternity under sperm competition in an external fertilizer

    Directory of Open Access Journals (Sweden)

    Torvald Blikra Egeland

    2015-07-01

    Full Text Available How much of a fitness benefit is obtained by dominant males of external fertilizers from releasing ejaculates in synchrony with female egg-release when engaging in sperm competition, and what is the most important sperm trait for paternity in these situations? The Arctic charr (Salvelinus alpinus is an external fertilizer experiencing intense male-male competition over reproductive opportunities including sperm competition. To compensate for their disadvantage the sneaker males, which often spawn out of synchrony with the female, produce more and faster sperm than the guarding males. We used controlled in vitro fertilization trials with experimentally produced dominant and subordinate, sneaker males to test what effect relative synchrony in gamete release, sperm quality (i.e., motility and velocity and sperm quantity have on a male’s fertilization success in pair-wise sperm competitions. When the sneaker males released ejaculates after the guarding male there was no overall difference in fertilization success. The quality (i.e., motility and velocity of a male’s sperm relative to that of the competing male was the best predictor of male fertilization success regardless of their mating tactic and spawning synchrony. The relative number of sperm cells also had an effect on fertilization success, but mainly when the dominant and sneaker male ejaculated synchronously. Our close imitation of natural sperm competition in charr shows that the sneaker males of external fertilizing species may fully compensate for their disadvantaged mating role by producing ejaculates of higher quality - an adjustment strangely not met by dominants.

  5. Synthesis and Phase Behavior of Poly(N-isopropylacrylamide)-b-Poly(L-Lysine Hydrochloride) and Poly(N-Isopropylacrylamide-co-Acrylamide)-b-Poly(L-Lysine Hydrochloride)

    NARCIS (Netherlands)

    Spasojevic, Milica; Vorenkamp, Eltjo; Jansen, Mark R. P. A. C. S.; de Vos, Paul; Schouten, Arend Jan

    The synthesis of poly(N-isopropylacrylamide)-b-poly(L-lysine) and poly(N-isopropylacrylamide- co-acrylamide)-b-poly(L-lysine) copolymers was accomplished by combining atom transfer radical polymerization (ATRP) and ring opening polymerization (ROP). For this purpose, a di-functional initiator with

  6. Molecular and structural insight into lysine selection on substrate and ubiquitin lysine 48 by the ubiquitin-conjugating enzyme Cdc34

    DEFF Research Database (Denmark)

    Suryadinata, Randy; Holien, Jessica K; Yang, George

    2013-01-01

    The attachment of ubiquitin (Ub) to lysines on substrates or itself by ubiquitin-conjugating (E2) and ubiquitin ligase (E3) enzymes results in protein ubiquitination. Lysine selection is important for generating diverse substrate-Ub structures and targeting proteins to different fates; however, t...

  7. Evaluation of methods to determine sperm density for the european eel, anguilla anguilla

    DEFF Research Database (Denmark)

    Sørensen, Sune Riis; Gallego, V.; Pérez, L.

    2013-01-01

    , computer-assisted sperm analysis (CASA) and flow cytometry (FCM), using Neubauer Improved haemocytometer as benchmark. Initially, relationships between spermatocrit, haemocytometer counts and sperm motility were analysed, as well as the effect of sperm dilution on haemocytometer counts. Furthermore...

  8. Sperm density required for inducing gynogenetic haploidy in scallop Chlamys nobilis

    Digital Repository Service at National Institute of Oceanography (India)

    Goswami, U.

    Experiments conducted for standardizing sperm density for artificial insemination and inducing gynogenesis by ultra-violet rays irradiated sperms in Chlamys nobilis showed that 10:1 ratio of sperms to eggs, gave maximum rate of fertilization...

  9. An update on sperm retrieval techniques for azoospermic males

    Directory of Open Access Journals (Sweden)

    Sandro C. Esteves

    2013-01-01

    Full Text Available The use of non-ejaculated sperm coupled with intracytoplasmic sperm injection has become a globally established procedure for couples with azoospermic male partners who wish to have biological offspring. Surgical methods have been developed to retrieve spermatozoa from the epididymides and the testes of such patients. This article reviews the methods currently available for sperm acquisition in azoospermia, with a particular focus on the perioperative, anesthetic and technical aspects of these procedures. A critical analysis of the advantages and disadvantages of these sperm retrieval methods is provided, including the authors' methods of choice and anesthesia preferences.

  10. Vitrification of neat semen alters sperm parameters and DNA integrity.

    Science.gov (United States)

    Khalili, Mohammad Ali; Adib, Maryam; Halvaei, Iman; Nabi, Ali

    2014-05-06

    Our aim was to evaluate the effect of neat semen vitrification on human sperm vital parameters and DNA integrity in men with normal and abnormal sperm parameters. Semen samples were 17 normozoospermic samples and 17 specimens with abnormal sperm parameters. Semen analysis was performed according to World Health Organization (WHO) criteria. Then, the smear was provided from each sample and fixed for terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining. Vitrification of neat semen was done by plunging cryoloops directly into liquid nitrogen and preserved for 7 days. The samples were warmed and re-evaluated for sperm parameters as well as DNA integrity. Besides, the correlation between sperm parameters and DNA fragmentation was assessed pre- and post vitrification. Cryopreserved spermatozoa showed significant decrease in sperm motility, viability and normal morphology after thawing in both normal and abnormal semen. Also, the rate of sperm DNA fragmentation was significantly higher after vitrification compared to fresh samples in normal (24.76 ± 5.03 and 16.41 ± 4.53, P = .002) and abnormal (34.29 ± 10.02 and 23.5 ± 8.31, P neat ejaculates has negative impact on sperm parameters as well as DNA integrity, particularly among abnormal semen subjects. It is, therefore, recommend to process semen samples and vitrify the sperm pellets.

  11. Average sperm count remains unchanged despite reduction in maternal smoking

    DEFF Research Database (Denmark)

    Priskorn, L; Nordkap, L; Bang, A K

    2018-01-01

    STUDY QUESTION: How are temporal trends in lifestyle factors, including exposure to maternal smoking in utero, associated to semen quality in young men from the general population? SUMMARY ANSWER: Exposure to maternal smoking was associated with lower sperm counts but no overall increase in sperm...... temporal trends. Parental age increased, and exposure in utero to maternal smoking declined from 40% among men investigated in 1996-2000 to 18% among men investigated in 2011-2016. Exposure to maternal smoking was associated with lower sperm counts but no overall increase in sperm counts was observed...

  12. Sex-sorting sperm using flow cytometry/cell sorting.

    Science.gov (United States)

    Garner, Duane L; Evans, K Michael; Seidel, George E

    2013-01-01

    The sex of mammalian offspring can be predetermined by flow sorting relatively pure living populations of X- and Y-chromosome-bearing sperm. This method is based on precise staining of the DNA of sperm with the nucleic acid-specific fluorophore, Hoechst 33342, to differentiate between the subpopulations of X- and Y-sperm. The fluorescently stained sperm are then sex-sorted using a specialized high speed sorter, MoFlo(®) SX XDP, and collected into biologically supportive media prior to reconcentration and cryopreservation in numbers adequate for use with artificial insemination for some species or for in vitro fertilization. Sperm sorting can provide subpopulations of X- or Y-bearing bovine sperm at rates in the 8,000 sperm/s range while maintaining; a purity of 90% such that it has been applied to cattle on a commercial basis. The sex of offspring has been predetermined in a wide variety of mammalian species including cattle, swine, horses, sheep, goats, dogs, cats, deer, elk, dolphins, water buffalo as well as in humans using flow cytometric sorting of X- and Y-sperm.

  13. Sperm quality in New Zealand: Is the downward trend continuing?

    Science.gov (United States)

    Birdsall, Mary A; Peek, John; Valiapan, Sumithra

    2015-10-16

    To investigate whether the decline in sperm concentration in New Zealand sperm donors observed from 1987 to 2007 continued in the period 2008-2014. A retrospective study from 2008 to 2014. The first semen sample of 285 men presenting as sperm donors in Auckland and Wellington was analysed for sperm concentration, seminal fluid volume and the percentage of motile sperm. These results were compared to results from 1987 to 2007 from the same clinics. The decline in semen volume and sperm concentration observed between 1987 and 2007 did not continue in 2008-2014. Sperm concentration decreased from 1987 until some time between 1997 and 2001, and has remained stable at an average of 62x106/ml between 2001 and 2014. Sperm motility declined significantly (8%) in the period 2008-2014, but there was no significant change over the total period studied, between 1987 and 2014. After a decline between 1987 and sometime during 1997-2001, the sperm concentration in men presenting as donors remained unchanged between 2002 and 2014, suggesting semen quality has not changed in New Zealand men over the last decade.

  14. Mass spectrometry profiling of oxysterols in human sperm identifies 25-hydroxycholesterol as a marker of sperm function

    Directory of Open Access Journals (Sweden)

    Chiara Zerbinati

    2017-04-01

    Full Text Available Cholesterol is a main lipid component of sperm cell that is essential for sperm membrane fluidity, capacitation, and acrosomal reaction. Recent data obtained in bovine sperm showed that sperm capacitation is associated to the formation of oxysterols, oxidized products of cholesterol. The aim of this study was to profile oxysterol content in human semen, and to investigate their potential role in sperm pathophysiology. Among the 12 oxysterols analyzed, 25-hydroxycholesterol (25-HC resulted the most represented in normozoospermic samples, and its concentration positively correlated with spermatozoa number. We detected Cholesterol 25-hydroxylase, the enzyme responsible for 25-HC production, in human spermatozoa at the level of the neck and the post acrosomal area. Upon incubation with spermatozoa, 25-HC induced calcium and cholesterol transients in connection with the acrosomal reaction. Our results support a role for 25-HC in sperm function.

  15. Mass spectrometry profiling of oxysterols in human sperm identifies 25-hydroxycholesterol as a marker of sperm function.

    Science.gov (United States)

    Zerbinati, Chiara; Caponecchia, Luisa; Puca, Rosa; Ciacciarelli, Marco; Salacone, Pietro; Sebastianelli, Annalisa; Pastore, Antonio; Palleschi, Giovanni; Petrozza, Vincenzo; Porta, Natale; Rago, Rocco; Carbone, Antonio; Iuliano, Luigi

    2017-04-01

    Cholesterol is a main lipid component of sperm cell that is essential for sperm membrane fluidity, capacitation, and acrosomal reaction. Recent data obtained in bovine sperm showed that sperm capacitation is associated to the formation of oxysterols, oxidized products of cholesterol. The aim of this study was to profile oxysterol content in human semen, and to investigate their potential role in sperm pathophysiology. Among the 12 oxysterols analyzed, 25-hydroxycholesterol (25-HC) resulted the most represented in normozoospermic samples, and its concentration positively correlated with spermatozoa number. We detected Cholesterol 25-hydroxylase, the enzyme responsible for 25-HC production, in human spermatozoa at the level of the neck and the post acrosomal area. Upon incubation with spermatozoa, 25-HC induced calcium and cholesterol transients in connection with the acrosomal reaction. Our results support a role for 25-HC in sperm function. Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.

  16. Sperm-surface ATP in boar spermatozoa is required for fertilization: relevance to sperm proteasomal function.

    Science.gov (United States)

    Yi, Young-Joo; Park, Chang-Sik; Kim, Eui-Sook; Song, Eun-Sook; Jeong, Ji-Hyeon; Sutovsky, Peter

    2009-01-01

    Extracellular ATP has been implicated in a number of cellular events, including mammalian sperm function. The complement of ATP-dependent sperm proteins includes six subunits of the 26S proteasome, a multi-subunit protease specific to ubiquitinated substrate-proteins. Proteolysis of ubiquitinated proteins by the 26S proteasome is necessary for the success of mammalian fertilization, including but not limited to acrosomal exocytosis (AE) and sperm-zona pellucida (ZP) penetration. The 26S proteasome is uniquely present on the sperm acrosomal surface during mammalian, ascidian, and invertebrate fertilization. The proteasome is a multi-subunit protease complex of approximately 2 MDa composed of the 19S regulatory complex and a 20S proteolytic core. Integrity of the 19S complex is maintained by six 19S ATPase subunits (PSMC1 through PSMC6). Consequently, we hypothesized that fertilization will be blocked by the depletion of sperm-surface associated ATP (ssATP). Depletion of ssATP by the Solanum tuberosum apyrase, a 49 kDa, non-cell permeant enzyme, significantly reduced the ATP content measured by an adapted luminescence-ATP assay from which all permeabilizing agents were excluded. Addition of active apyrase to porcine in vitro fertilization (IVF) medium caused a concentration dependent reduction in the overall fertilization rate. No such outcomes were observed in control groups using heat-inactivated apyrase. Apyrase treatment altered the band pattern of 19S ATPase subunits PSMC1 (Rpt2) and PSMC4 (Rpt3) in Western blotting, suggesting that it had an effect on the integrity of the sperm proteasomal 19S complex. Apyrase only altered the proteasomal core activities slightly, since these activities are not directly dependent on external ATP. In contrast, sperm treatment with MG132, a specific inhibitor of the proteasomal core chymotrypsin-like activity, inhibited the target proteolytic activity, but also induced a compensatory elevation in proteasomal peptidyl

  17. Honey bee males and queens use glandular secretions to enhance sperm viability before and after storage

    DEFF Research Database (Denmark)

    Den Boer, Susanne Petronella A; Boomsma, Jacobus Jan; Baer, Boris

    2009-01-01

    on sperm survival. After sperm storage, the queens also provide sperm with secretions from spermathecal glands and we show that these secretions have a comparable positive effect on sperm viability. SDS gels show that the proteomic profiles of accessory gland secretion and spermathecal fluid secretion...... hardly overlap, which suggests that males and females use different proteins to enhance sperm viability during, respectively, ejaculation and final sperm storage....

  18. Evidence for biased use of sperm sources in wild female giant cuttlefish (Sepia apama)

    OpenAIRE

    Naud, Marie-José; Shaw, Paul W; Hanlon, Roger T; Havenhand, Jon N

    2005-01-01

    In species where females store sperm from their mates prior to fertilization, sperm competition is particularly probable. Female Sepia apama are polyandrous and have access to sperm from packages (spermatangia) deposited by males onto their buccal area during mating and to sperm stored in internal sperm-storage organs (receptacles) located below the beak. Here, we describe the structure of the sperm stores in the female's buccal area, use microsatellite DNA analyses to determine the genetic d...

  19. Sperm survival in the female reproductive tract in the fly Scathophaga stercoraria (L.)

    OpenAIRE

    Bernasconi, G.; Hellriegel, B.; Heyland, A.; Ward, P. I.

    2002-01-01

    While sperm competition risk favours males transferring many sperm to secure fertilizations, females of a variety of species actively reduce sperm numbers reaching their reproductive tract, e.g. by extrusion or killing. Potential benefits of spermicide to females include nutritional gains, influence over sperm storage and paternity, and the elimination of sperm bearing somatic mutations that would lower zygote fitness.We investigated changes in sperm viability after in vivo and in vitro expos...

  20. Application of artificial insemination technique to eupyrene and/or apyrene sperm in Bombyx mori

    OpenAIRE

    Sahara, Ken; Takemura, Yoko

    2003-01-01

    The silkworm, Bombyx mori, has a dimorphic sperm system. The eupyrene sperm is the sperm to fertilize eggs and the apyrene sperm plays a crucial role for assisting fertilization. Heat-treated (331C for 96h) Daizo (DH) males, one of the strains in the silkworm, produce only eupyrene sperm, while in triploid males only apyrene sperm are functional. Though both types of males are found to be sterile, double copulation of the two males with a single female greatly increases fertili...

  1. Implementing an open-access CASA software for the assessment of stallion sperm motility: Relationship with other sperm quality parameters.

    Science.gov (United States)

    Giaretta, Elisa; Munerato, Mauro; Yeste, Marc; Galeati, Giovanna; Spinaci, Marcella; Tamanini, Carlo; Mari, Gaetano; Bucci, Diego

    2017-01-01

    Setting an open-access computer assisted sperm analysis (CASA) may benefit the evaluation of motility in mammalian sperm, especially when economic constraints do not allow the use of a commercial system. There have been successful attempts to develop such a device in Zebra fish sperm and the system has been used in very few studies on mammalian spermatozoa. Against this background, the present study aimed at developing an open-access CASA system for mammalian sperm using the horse as a model and based upon the Image J software previously established for Zebra fish sperm. Along with determining the sperm progressive motility and other kinetic parameters (such as amplitude of lateral head displacement), the "results" window was adjusted to simplify subsequent statistical analyses. The path window was enriched with colored sperm trajectories on the basis of the subpopulation they belong to and a number that allowed the sperm track to be associated to the sperm motility data shown in the "results" window. Data obtained from the novel plugin (named as CASA_bgm) were compared with those of the commercial CASA Hamilton-Thorn IVOS Vers.12, through Bland Altman's plots. While the percentage of total and progressive motile sperm, VCL, VAP, VSL, LIN and STR and ALH were in agreement with those obtained with the commercial system, BCF significantly differed between the two systems probably due to their settings. Interestingly, a positive and significant correlation between the percentages of total motile sperm evaluated through CASA_bgm and those showing high mitochondrial membrane potential evaluated by JC-1 staining was found. In conclusion, CASA_bgm ImageJ plugin could be useful and reliable for stallion sperm motility analysis and it is our aim to apply this system to other mammalian species. Copyright © 2016 Elsevier B.V. All rights reserved.

  2. Anti-allergic effects of a nonameric peptide isolated from the intestine gastrointestinal digests of abalone (Haliotis discus hannai) in activated HMC-1 human mast cells.

    Science.gov (United States)

    Ko, Seok-Chun; Lee, Dae-Sung; Park, Won Sun; Yoo, Jong Su; Yim, Mi-Jin; Qian, Zhong-Ji; Lee, Chang-Min; Oh, Junghwan; Jung, Won-Kyo; Choi, Il-Whan

    2016-01-01

    The aim of the present study was to examine whether the intestine gastrointestinal (GI) digests of abalone [Haliotis discus hannai (H. discus hannai)] modulate inflammatory responses and to elucidate the mechanisms involved. The GI digests of the abalone intestines were fractionated into fractions I (>10 kDa), II (5-10 kDa) and Ⅲ (<5 kDa). Of the abalone intestine GI digests (AIGIDs), fraction Ⅲ inhibited the passive cutaneous anaphylaxis (PCA) reaction in mice. Subsequently, a bioactive peptide [abalone intestine GI digest peptide (AIGIDP)] isolated from fraction Ⅲ was determined to be 1175.2 Da, and the amino acid sequence was found to be PFNQGTFAS. We noted that the purified nonameric peptide (AIGIDP) attenuated the phorbol‑12‑myristate 13-acetate plus calcium ionophore A23187 (PMACI)-induced histamine release and the production of pro-inflammatory cytokines, such as tumor necrosis factor-α (TNF-α), interleukin (IL)-1β and IL-6 in human mast cells (HMC-1 cells). In addition, we also noted that AIGIDP inhibited the PMACI‑induced activation of nuclear factor‑κB (NF-κB) by suppressing IκBα phosphorylation and that it suppressed the production of cytokines by decreasing the phosphorylation of JNK. The findings of our study indicate that AIGIDP exerts a modulatory, anti-allergic effect on mast cell-mediated inflammatory diseases.

  3. First characterisation of the populations and immune-related activities of hemocytes from two edible gastropod species, the disk abalone, Haliotis discus discus and the spiny top shell, Turbo cornutus.

    Science.gov (United States)

    Donaghy, Ludovic; Hong, Hyun-Ki; Lambert, Christophe; Park, Heung-Sik; Shim, Won Joon; Choi, Kwang-Sik

    2010-01-01

    The disk abalone Haliotis discus discus and the spiny top shell Turbo cornutus are edible gastropod species of high economic value, mainly in Asia. Mortality outbreaks and variations in worldwide stock abundance have been reported and suggested to be associated, at least in part, with pathogenic infections. Ecology, biology and immunology of both species are currently not well documented. The characterisation of the immune systems of these species is necessary to further assess the responses of H. discus discus and T. cornutus to environmental, chemical and disease stresses. In the present study, we investigated the morphology and immune-related activities of hemocytes in both species using light microscopy and flow cytometry. Two types of hemocytes were identified in the disk abalone hemolymph, blast-like cells and hyalinocytes; whereas four main hemocyte types were distinguished in the spiny top shell, blast-like cells, type I and II hyalinocytes, and granulocytes. Flow cytometric analysis also revealed differences between cell types in immune-related activities. Three subsets of hemocytes, defined by differing lysosomal characteristics, were observed in the hemolymph of the spiny top shell, and only one in the disk abalone. Phagocytic activity was higher in H. discus discus hemocytes than in T. cornutus hemocytes, and the kinetics of PMA-stimulated oxidative activity was different between hemocytes of the disk abalone and the spiny top shell. Finally our results suggest for the first time a predominant mitochondrial origin of oxidative activity in gastropod hemocytes. Copyright 2009 Elsevier Ltd. All rights reserved.

  4. Seasonal variation of sperm quality and the relationship between spermatocrit and sperm concentration in yamú Brycon amazonicus

    DEFF Research Database (Denmark)

    Cruz-Casallas, Pablo E.; Medina-Robles, Víctor M.; Velasco-Santamaría, Yohana M.

    2007-01-01

    and sperm concentration were examined over two reproductive seasons. Activation time, spermatocrit, and sperm concentration were significantly higher in 2003 than in 2004. Spermatocrit and sperm concentration showed a significant positive relationship (r2=0.79). During the 2004 reproductive season......, a significant variation was observed for spermatocrit and sperm concentration between the initial period (February 15th to March 14th) and the middle and final periods (March 15th to April 14th and April 15th to May 14th, respectively). Neither the ion concentrations nor the glucose and cholesterol...

  5. Flow cytometric sexing of spider sperm reveals an equal sperm production ratio in a female-biased species

    DEFF Research Database (Denmark)

    Vanthournout, Bram; Deswarte, K; Hammad, H

    2014-01-01

    that other factors influence sex ratio variation. In this paper, we investigate whether this additional variation can be explained by the unequal production of male- and female-determining sperm cells during sperm production. Using flow cytometry, we show that males produce equal amounts of male- and female......-determining sperm cells; thus bias in sperm production does not contribute to the sex ratio bias observed in this species. This demonstrates that other factors such as parental genes suppressing endosymbiont effects and cryptic female choice might play a role in sex allocation in this species....

  6. Effect of postthaw storage time and sperm-to-egg ratio on fertility of cryopreserved brook trout sperm.

    Science.gov (United States)

    Nynca, J; Dietrich, G J; Dobosz, S; Zalewski, T; Ciereszko, A

    2015-01-15

    The aim of this study was to test the influence of postthaw storage time on sperm motility parameters of brook trout (n = 9). Furthermore, we examined the effect of sperm-to-egg ratios of 300,000:1 and 600,000:1 on fertility of postthaw, cryopreserved, brook trout sperm. The application of a cryopreservation procedure produced very high postthaw sperm motility (56.8 ± 4.0%). The cryopreserved sperm of brook trout could be stored up to 60 minutes without loss of the percentage of sperm motility (52.0 ± 9.0%). The fertilization capacity of brook trout postthaw sperm was comparable with the fertilization rate of fresh semen at a sperm-to-egg ratio as low as 300,000:1 (42.4 ± 14.0% and 36.5 ± 11.0% for eyed and hatched stages, respectively). The possibility of postthaw semen storage for the prolonged time and the obtainment of high fertilization rate at low sperm-to-egg ratio can lead to the significant improvement of brook trout semen cryopreservation procedure. Copyright © 2015 Elsevier Inc. All rights reserved.

  7. Sperm-egg recognition in the mouse: characterization of sp56, a sperm protein having specific affinity for ZP3.

    Science.gov (United States)

    Cheng, A; Le, T; Palacios, M; Bookbinder, L H; Wassarman, P M; Suzuki, F; Bleil, J D

    1994-05-01

    Recognition between mammalian gametes occurs when the plasma membrane of the sperm head binds to the zona pellucida (ZP), an extracellular coat surrounding eggs. ZP3, one of three glycoproteins in the ZP, is the egg protein recognized by sperm. A mouse sperm surface protein, sp56 (M(r) = 56,000), has been identified on the basis of its specific affinity for ZP3 (Bleil, J. D., and P. M. Wassarman. 1990. Proc. Natl. Acad. Sci. USA. 87:5563-5567). Studies presented here were designed to characterize mouse sperm sp56 and to further test whether or not this protein specifically recognizes ZP3. sp56 was purified by both ZP3 affinity chromatography and by ion exchange chromatography followed by size-exclusion chromatography. The purified native protein eluted from size-exclusion columns as a homomultimer (M(r) approximately 110,000). Each monomer of the protein contains intramolecular disulfide bonds, consistent with its extracellular location. Immunohistochemical and immunoblotting studies, using monoclonal antibodies, demonstrated that sp56 is a peripheral membrane protein located on the outer surface of the sperm head plasma membrane, precisely where sperm bind ZP3. Results of crosslinking experiments demonstrated that the ZP3 oligosaccharide recognized by sperm has specific affinity for sp56. Collectively, these results suggest that sp56 may be the sperm protein responsible for sperm-egg recognition in the mouse.

  8. Methodical investigations on the determination of metabolic lysine requirements in broiler chickens. 1

    International Nuclear Information System (INIS)

    Bergner, H.; Nguyen Thi Nhan; Wilke, A.

    1987-01-01

    For the estimation of lysine requirement 128 male broiler chickens were used at an age of 7 to 21 days posthatching. They received a lysine-deficient diet composed of wheat and wheat gluten. To this basal diet L-lysine-HCL was supplemented successively resulting in 8 lysine levels ranging from 5.8 to 23.3 g lysine per kg dry matter (DM) (2.2 to 8.7 g lysine per 16 g N). At the end of the two-week feeding period of the experimental diets 14 C-lysine was injected intravenously 1.5 and 5.5 hours after feed withdrawal. During the following 4 hours the exretion of CO 2 and 14 CO 2 was measured. The highest daily gain of 21.5 g was observed in animals fed 13.3 g lysine-kg DM. Lysine concentrations exceeding 18.3 g/kg DM depressed body weight gain. The CO 2 excretion was not influenced by lysine intake. 14 CO 2 excretion was low with diets low in lysine content and increased 3 to 4 times with diets meeting the lysine requirement. Based on measurements 1.5 to 5.5 hours after feed withdrawal the saturation value for lysine was reached at 13.3 g/kg DM. This value was lowered (10.8 g/kg DM), however, if the estimation was carried out 5.5 to 9.5 hours after feed withdrawal. These results suggest a higher metabolic lysine requirement during the earlier period after feed intake. Both, reduced weight gain and non linearity in 14 CO 2 excretion in diets exceeding a lysine content of 18.3 g/kg DM indicate a limited capacity of the organism to degrade excessive lysine. According to the results a lysine requirement betwen 10.8 and 13.3 g/kg DM (27% CP and 660 EFU/sub hen//kg DM) was estimated for broiler chickens 3 weeks posthatching. (author)

  9. The predictive value of parameters of clinical presentations for sperm yield in patients with nonobstructive azoospermia receiving microdissection testicular sperm extraction

    Directory of Open Access Journals (Sweden)

    Ming-Hsuan Ku

    2017-12-01

    Conclusion: Clinical presentations or phenotypes can be used as predictive factors for successful sperm retrieval during mTESE in patients with NOA. Hypogonadotropic hypogonadism and cases with UDT history have a higher chance of sperm retrieval. Initial testicular needle biopsy, if available, can provide valuable information about chances of sperm retrieval. Hypospermatogenesis predicts high sperm yield rate, and LMA can have best upgrade results of sperm yield after mTESE.

  10. Air pollution and quality of sperm: a meta-analysis.

    Science.gov (United States)

    Fathi Najafi, Tahereh; Latifnejad Roudsari, Robab; Namvar, Farideh; Ghavami Ghanbarabadi, Vahid; Hadizadeh Talasaz, Zahra; Esmaeli, Mahin

    2015-04-01

    Air pollution is common in all countries and affects reproductive functions in men and women. It particularly impacts sperm parameters in men. This meta-analysis aimed to examine the impact of air pollution on the quality of sperm. The scientific databases of Medline, PubMed, Scopus, Google scholar, Cochrane Library, and Elsevier were searched to identify relevant articles published between 1978 to 2013. In the first step, 76 articles were selected. These studies were ecological correlation, cohort, retrospective, cross-sectional, and case control ones that were found through electronic and hand search of references about air pollution and male infertility. The outcome measurement was the change in sperm parameters. A total of 11 articles were ultimately included in a meta-analysis to examine the impact of air pollution on sperm parameters. The authors applied meta-analysis sheets from Cochrane library, then data extraction, including mean and standard deviation of sperm parameters were calculated and finally their confidence interval (CI) were compared to CI of standard parameters. The CI for pooled means were as follows: 2.68 ± 0.32 for ejaculation volume (mL), 62.1 ± 15.88 for sperm concentration (million per milliliter), 39.4 ± 5.52 for sperm motility (%), 23.91 ± 13.43 for sperm morphology (%) and 49.53 ± 11.08 for sperm count. The results of this meta-analysis showed that air pollution reduces sperm motility, but has no impact on the other sperm parameters of spermogram.

  11. Clinical aspects of sperm DNA fragmentation detection and male infertility.

    Science.gov (United States)

    Evenson, Donald P; Wixon, Regina

    2006-03-15

    Over the past 25 years, various methods have been developed to measure sperm DNA strand breaks in situ. Currently, there are four major tests of sperm DNA fragmentation, including the Comet, Tunel, sperm chromatin structure assay (SCSA) and the acridine orange test (AOT). The Comet assay is a light microscope technique where the sperm cells are mixed with melted agarose and then placed on a glass slide. The cells are lysed and then subjected to horizontal electrophoresis. The Tunel assay, another light microscope technique, transfers labeled nucleotide to the 3'OH group of a broken DNA strand with the use of terminal deoxynucleotidyl transferase. The fluorescence intensity of each scored sperm is determined as a "yes" or "no" for sperm on a light microscope slide or by channels of fluorescent intensity in a flow cytometer. The light microscope-based AOT, uses the metachromatic properties of acridine orange to stain sperm cells. The SCSA treats sperm with low pH to denature DNA at the sites of DNA strand breaks, followed by acridine orange (AO) staining of green for native DNA and red for denatured DNA as measured by flow cytometry (FCM) as well as % sperm with high DNA stainability (HDS: immature sperm with intact DNA related to decreased fertilization rates). The SCSA method has defined a 27-30% DNA fragmentation index (DFI) as the point in which a man is placed into a statistical category of taking a longer time to in vivo pregnancy, intra uterine insemination (IUI) and more routine in vitro fertilization (IVF) cycles or no pregnancy. Current data suggest that intracytoplasmic sperm injection (ICSI) may help overcome the diminished pregnancy prognosis with high DFI over the other ART or natural methods.

  12. Rosiglitazone Improves Stallion Sperm Motility, ATP Content, and Mitochondrial Function.

    Science.gov (United States)

    Swegen, Aleona; Lambourne, Sarah Renay; Aitken, R John; Gibb, Zamira

    2016-11-01

    Media used for equine sperm storage often contain relatively high concentrations of glucose, even though stallion spermatozoa preferentially utilize oxidative phosphorylation (OXPHOS) over glycolysis to generate ATP and support motility. Rosiglitazone is an antidiabetic compound that enhances metabolic flexibility and glucose utilization in various cell types, but its effects on sperm metabolism are unknown. This study investigated the effects of rosiglitazone on stallion sperm function in vitro, along with the possible role of AMP-activated protein kinase (AMPK) in mediating these effects. Spermatozoa were incubated with or without rosiglitazone, GW9662 (an antagonist of peroxisome proliferator-activating receptor-gamma), and compound C (CC; an AMPK inhibitor). Sperm motility, viability, reactive oxygen species production, mitochondrial membrane potential (mMP), ATP content, and glucose uptake capacity were measured. Samples incubated with rosiglitazone displayed significantly higher motility, percentage of cells with normal mMP, ATP content, and glucose uptake capacity, while sperm viability was unaffected. The percentage of spermatozoa positive for mitochondrial ROS was also significantly lower in rosiglitazone-treated samples. AMPK localized to the sperm midpiece, and its phosphorylation, was increased in rosiglitazone-treated spermatozoa. CC decreased sperm AMPK phosphorylation and reduced sperm motility, and successfully inhibited the effects of rosiglitazone. Inclusion of rosiglitazone in a room temperature sperm storage medium maintained sperm motility above 60% for 6 days, attaining significantly higher motility than sperm stored in control media. The ability of rosiglitazone to substantially alleviate the time-dependent deterioration of stallion spermatozoa by diverting metabolism away from OXPHOS and toward glycolysis has novel implications for the long-term, functional preservation of these cells. © 2016 by the Society for the Study of Reproduction

  13. Pentoxifylline effects on capacitation and fertility of stallion epididymal sperm.

    Science.gov (United States)

    Guasti, P N; Monteiro, G A; Maziero, R R D; Carmo, M T; Dell'Aqua, J A; Crespilho, A M; Rifai, E A; Papa, F O

    2017-04-01

    The aims of this study were to determinate whether pentoxifylline (PTX) increases the motion parameters of fresh and frozen-thawed equine epididymal spermatozoa, to evaluate the tyrosine phosphorylation of frozen-thawed epididymal sperm in the presence of PTX and to determine whether the PTX-treatment of stallion epididymal sperm prior to freezing improves the fertility response of mares to a reduced number of spermatozoa per insemination dose. Fifty epididymis were flushed with a skim milk based extender with or without PTX. The pre-treatment with PTX enhanced the sperm motility after being harvested (Psperm kinematics between control and treated samples (P>0.05). Plasma membrane integrity did not differ between control and PTX group after recovery and after thawing (P>0.05), as observed in tyrosine phosphorylation, which the PTX treatment did not alter the percentage of tail-associated immunofluorescence of cryopreserved epididymal sperm (P>0.05). For the fertility trial, different insemination groups were tested: 800×10 6 epididymal sperm (C800); 100×10 6 epididymal sperm (C100); 100×10 6 epididymal sperm recovered in an extender containing PTX (PTX100). The conception rates for C800; C100 and PTX100 were 68.7% (11/16); 31.5% (5/16) and 50% (8/16), respectively. The conception rate did not differ among groups (P>0.05), however, a low number of animals was used in this study. A trend toward significance (P=0.07) was observed between C800 and C100 groups. In conclusion, PTX has no deleterious effect on sperm motility, viability and capacitation of cryopreserved stallion epididymal sperm. The conventional artificial insemination with 100×10 6 sperm recovered with PTX ensures acceptable conception rates and maximize the limited number of doses of cryopreserved stallion epididymal sperm. Copyright © 2017 Elsevier B.V. All rights reserved.

  14. Conformational Studies of ε- CBz- L- Lysine and L- Valine Block Copolypeptides

    Directory of Open Access Journals (Sweden)

    Ajay Kumar

    2010-01-01

    Full Text Available Conformational studies of ε-CBz-L-lysine and L-valine block copoylpeptides using x- ray diffraction and CD spectra are described. The block copolypeptides contain valine block in the center and on both side of the valine are ε-CBz-L-lysine blocks. The conformation of the copolypeptides changes with increases in the chain length of ε- CBz-L- lysine blocks. When length of ε- CBZ- L- lysine blocks is 9, the block copolypeptide has exclusive beta sheet structure. With the increase in chain length of ε-CBz-L-lysine blocks from 9 to 14, the block copolypeptide shows presence of both alpha helix and beta sheet components. With further increase in chain length of ε- CBz- L- lysine blocks, the beta sheet component disappears and block copolypeptides exhibits exclusive α -helix conformation.

  15. Molecular characterization of collagen IV evidences early transcription expression related to the immune response against bacterial infection in the red abalone (Haliotis rufescens).

    Science.gov (United States)

    Chovar-Vera, Ornella; Valenzuela-Muñoz, Valentina; Gallardo-Escárate, Cristian

    2015-02-01

    Collagen IV has been described as a structural protein of the basement membrane, which as a whole forms a specialized extracellular matrix. Recent studies have indicated a possible relationship between collagen IV and the innate immune response of invertebrate organisms. The present study characterized the alpha-1 chain of collagen IV in the red abalone Haliotis rufescens (Hr-ColIV) and evaluated its association with the innate immune response against Vibrio anguillarum. To further evidence the immune response, the matrix metalloproteinase-1 (Hr-MMP-1) and C-type lectin (Hr-CLEC) genes were also assessed. The complete sequence of Hr-ColIV was composed of 6658 bp, with a 5'UTR of 154 bp, a 3'UTR of 1177 bp, and an ORF of 5327 bp that coded for 1776 amino acids. The innate immune response generated against V. anguillarum resulted in a significant increase in the transcript levels of Hr-ColIV between 3 and 6 hpi, whereas Hr-MMP-1 and Hr-CLEC had the highest transcript activity 6 and 12 hpi, respectively. The results obtained in this study propose a putative biological function for collagen IV involved in the early innate immune response of the red abalone H. rufescens. Copyright © 2014 Elsevier Ltd. All rights reserved.

  16. HdhCTL1 is a novel C-type lectin of abalone Haliotis discus hannai that agglutinates Gram-negative bacterial pathogens.

    Science.gov (United States)

    Zhang, Jian; Qiu, Reng; Hu, Yong-hua

    2014-12-01

    C-type lectins (CTLs) are Ca(2+)-dependent carbohydrate recognition proteins, which play important roles in the innate immunity of both vertebrates and invertebrates. In this study, we identified and characterized a C-type lectin (named HdhCTL1) from Pacific abalone, Haliotis discus hannai. HdhCTL1 is composed of 176 amino acid residues and shares low (23.9%) identity with the known CTL of abalone. HdhCTL1 possesses a putative signal peptide and a carbohydrate-recognition domain (CRD) typical of CTLs. The CRD of HdhCTL1 contains four disulfide bond-forming cysteine residues that are highly conserved in CTLs. HdhCTL1 mRNA was detected in a wide range of tissues and expressed abundantly in the digestive gland. Experimental infection with the bacterial pathogen Vibrio anguillarum significantly upregulated HdhCTL1 expression in a time-dependent manner. Recombinant HdhCTL1 (rHdhCTL1) purified from Escherichia coli was able to agglutinate Gram-negative bacterial pathogens. The agglutinating ability of rHdhCTL1 was abolished in the presence of mannose. These results suggest that HdhCTL1 is a novel CTL which is likely to be involved in host defense against bacterial infection. Copyright © 2014 Elsevier Ltd. All rights reserved.

  17. Molecular characterization and mRNA expression during metal exposure and thermal stress of copper/zinc- and manganese-superoxide dismutases in disk abalone, Haliotis discus discus.

    Science.gov (United States)

    Kim, Keun-Yong; Lee, Sang Yoon; Cho, Young Sun; Bang, In Chul; Kim, Ki Hong; Kim, Dong Soo; Nam, Yoon Kwon

    2007-11-01

    Complementary DNAs encoding copper/zinc superoxide dismutase (Cu/Zn-SOD; SOD1) and manganese superoxide dismutase (Mn-SOD; SOD2) were isolated from disk abalone, Haliotis discus discus. The open reading frame sequences of Cu/Zn- and Mn-SODs encoded 154 and 226 amino acids, respectively. Multiple sequence alignments using the deduced amino acid sequences revealed that both abalone SODs showed considerable sequence similarities with their orthologues from diverse aerobic organisms, in which the amino acid residues forming metal ligands were highly conserved. All phylogenetic trees for both SOD genes inferred from maximum likelihood and Bayesian inference analyses presented the monophyletic status of Teleostei and Aves/Tetrapoda clades, and recovered relatively close genetic affiliation of H. discus discus with some molluscan species. Expression of both SODs at mRNA levels were highly modulated in various tissues (gill, muscle and hepatopancreas from juveniles, and haemocytes from adults) by experimental exposures to heavy metals (copper, zinc and cadmium) and also by thermal treatments (elevation of temperature). The mRNA levels of both SODs were increased in general during the metal or thermal treatments; however, the transcriptional responses of SOD genes were quite variable depending upon isoforms and tissues based on semi-quantitative and/or real-time RT-PCR assays.

  18. Ultrastructural description of new Rickettsia-like organisms in the commercial abalone Haliotis tuberculata (Gastropoda: Haliotidae) from the NW of Spain.

    Science.gov (United States)

    Azevedo, Carlos; Conchas, Ramón F; Tajdari, Jessica; Montes, Jaime

    2006-08-30

    Rickettsia-like organisms (RLOs) were found in the commercially farmed abalone Haliotis tuberculata in the northwestern region of the Atlantic Coast of Spain and are described from light and transmission electron microscopy observations. The RLOs measured approximately 1.6 x 0.9 microm and were found in intracytoplasmic, spherical to ellipsoidal vacuoles (up to 8 microm) in the epithelial cells of the digestive diverticulae. The morphological ultrastructure of these organisms was typically prokaryotic, including a plasmalemma and a thin Gram-negative type cell wall. Several ultrastructural changes were observed in the epithelial cells of the host containing the RLOs. The nuclei became pycnotic and several basophilic dense inclusions appeared in the cytoplasm. In addition, the host cell appeared lysed and was ruptured in advanced stages of infection. It was impossible to ascertain whether the RLOs are responsible for this disease, as a haplosporidian infection was also present. We can only conclude that the presence of RLOs simultaneously with a haplosporidian parasite may contribute to the mortality of the abalone host.

  19. Coupling sperm mediated gene transfer and sperm sorting techniques: a new perspective for swine transgenesis.

    Science.gov (United States)

    De Cecco, Marco; Spinaci, Marcella; Zannoni, Augusta; Bernardini, Chiara; Seren, Eraldo; Forni, Monica; Bacci, Maria Laura

    2010-09-15

    Flow cytometric separation of X and Y chromosome-bearing spermatozoa has been demonstrated to be effective in pigs, allowing the use of boar sexed semen in in vitro trials. Sperm Mediated Gene Transfer (SMGT) is a widely used and efficient technique for the creation of transgenic animals. The present research intended to prove that it is possible to associate sperm sexing with the SMGT technique in order to speed up the assessment of homozygous lines of transgenic pigs. In the first experiment, the sorting protocol was modified in order to obtain the highest DNA uptake by sorted spermatozoa. In the second experiment, spermatozoa that had undergone only sperm sorting, only SMGT, or both procedures (Sorted-SMGT) were used for in in vitro fertilization of in vitro matured oocytes. In the third experiment, transformed blastocysts of the desired gender (male) were obtained with Sorted-SMGT in an in vitro fertilization trial. The method we developed here allowed us to produce transgenic swine blastocysts of pre-determined gender, giving a positive answer at the aim to couple SMGT and sperm sorting in swine, obtaining fertile spermatozoa able to produce transgenic embryos of pre-determined gender. Copyright 2010 Elsevier Inc. All rights reserved.

  20. Birefringence characteristics in sperm heads allow for the selection of reacted spermatozoa for intracytoplasmic sperm injection.

    Science.gov (United States)

    Gianaroli, Luca; Magli, M Cristina; Ferraretti, Anna P; Crippa, Andor; Lappi, Michela; Capitani, Serena; Baccetti, Baccio

    2010-02-01

    To verify clinical outcome after injection of spermatozoa that have undergone the acrosome reaction (reacted spermatozoa) vs. those still having an intact acrosome (nonreacted spermatozoa). Prospective, randomized study. Reproductive Medicine Unit, Italian Society for the Study of Reproductive Medicine, Bologna, Italy. According to a prospective randomization including 71 couples with severe male factor infertility, intracytoplasmic sperm injection (ICSI) was performed under polarized light that permitted analysis of the pattern of birefringence in the sperm head. Twenty-three patients had their oocytes injected with reacted spermatozoa, 26 patient's oocytes were injected with nonreacted spermatozoa, and in 22 patients both reacted and nonreacted spermatozoa were injected. Intracytoplasmic sperm injection was performed under polarized light to selectively inject acrosome-reacted and acrosome-nonreacted spermatozoa. Rates of fertilization, cleavage, pregnancy, implantation, and ongoing implantation. There was no effect on the fertilizing capacity and embryo development of either type of sperm, whereas the implantation rate was higher in oocytes injected with reacted spermatozoa (39.0%) vs. those injected with nonreacted spermatozoa (8.6%). The implantation rate was 24.4% in the group injected with both reacted and nonreacted spermatozoa. The delivery rate per cycle followed the same trend. Spermatozoa that have undergone the acrosome reaction seem to be more prone to supporting the development of viable ICSI embryos. Copyright 2010 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  1. Towards microfluidic sperm refinement : impedance-based analysis and sorting of sperm cells

    NARCIS (Netherlands)

    de Wagenaar, B.; Dekker, Stefan; de Boer, Hans L.; Bomer, Johan G.; Olthuis, Wouter; van den Berg, Albert; Segerink, Loes Irene

    2016-01-01

    The use of high quality semen for artificial insemination in the livestock industry is essential for successful outcome. Insemination using semen with a high number of sperm cells containing morphological defects has a negative impact on fertilization outcome. Therefore, semen with a high number of

  2. Cryopreservation of European catfish Silurus glanis sperm: Sperm motility, viability and hatching success of embryos

    Czech Academy of Sciences Publication Activity Database

    Linhart, Otomar; Rodina, Marek; Flajšhans, Martin; Gela, David; Kocour, Martin

    2005-01-01

    Roč. 51, č. 3 (2005), s. 250-261 ISSN 0011-2240 R&D Projects: GA ČR GA524/03/0178; GA AV ČR IBS5045314 Institutional research plan: CEZ:AV0Z50450515 Keywords : fish * sperm * aquaculture Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 1.765, year: 2005

  3. Sperm maturation in dogs: sperm profile and enzymatic antioxidant status in ejaculated and epididymal spermatozoa.

    Science.gov (United States)

    Angrimani, D S R; Lucio, C F; Veiga, G A L; Silva, L C G; Regazzi, F M; Nichi, M; Vannucchi, C I

    2014-09-01

    Spermatozoa become more susceptible to the attack of reactive oxygen species during maturation. To avoid oxidative damage, the epididymis must provide the necessary antioxidant protection. The aim of this study was to compare the canine sperm profile and the enzymatic antioxidant status of the ejaculated fractions and samples collected from the different segments of the epididymis (caput, corpus and cauda). Five adult dogs were used, and after 1-3 weeks, subsequently to bilateral orchiectomy and epididymal storage, sperm samples were collected from the different segments of the epididymis. Samples were evaluated for conventional microscopy and computer-assisted motility analysis: sperm plasma membrane permeability and the activity of the antioxidant enzymes catalase, glutathione peroxidase (GPx) and superoxide dismutase (SOD). Samples collected from the caput and corpus showed lower values for most of the motility variables evaluated, indicating different levels of immaturity. Catalase activity was observed only in ejaculated samples. Conversely, GPx activity was higher in the cauda epididymidis. Correlations were found between SOD and GPx and SOD and sperm motility in the epididymal cauda and corpus, highlighting the importance of the enzymes for the protection of spermatozoa during the transit along the epididymis. © 2013 Blackwell Verlag GmbH.

  4. Antimicrobial activity of bovine NK-lysin-derived peptides on bovine respiratory pathogen Histophilus somni

    OpenAIRE

    Dassanayake, Rohana P.; Falkenberg, Shollie M.; Briggs, Robert E.; Tatum, Fred M.; Sacco, Randy E.

    2017-01-01

    Bovine NK-lysins, which are functionally and structurally similar to human granulysin and porcine NK-lysin, are predominantly found in the granules of cytotoxic T-lymphocytes and NK-cells. Although antimicrobial activity of bovine NK-lysin has been assessed for several bacterial pathogens, not all the important bacterial pathogens that are involved in the bovine respiratory disease complex have been studied. Therefore the objective of the present study was to evaluate the antimicrobial activi...

  5. [Artificial insemination with donor sperm in the Netherlands: future-proof?].

    Science.gov (United States)

    Kop, P A L Femke; Janssens, Pim M W; Mochtar, Monique H

    2014-01-01

    In recent years much has changed in care for artificial insemination with donor sperm (AID). Since new laws and regulations were implemented, a large number of sperm banks have closed and the total number of sperm donors and their availability have decreased. Long waiting times and the use of sperm donors recruited by foreign commercial sperm banks can indicate a shortage of sperm donors. The fact that the internet offers women the possibility of ordering donor sperm and starting treatment without the intervention of a sperm bank means that future donor-conceived children may be prevented from obtaining the identity of their sperm donor as stipulated in the Dutch law on donor information in the context of artificial insemination. In order to comply with this law, an active recruitment policy is needed for Dutch sperm donors, to prevent waiting lists and treatments outside Dutch sperm banks. Only then can current AID care be guaranteed in the Netherlands in the future.

  6. Beyond Histones: New Substrate Proteins of Lysine Deacetylases in Arabidopsis Nuclei

    Directory of Open Access Journals (Sweden)

    Magdalena Füßl

    2018-04-01

    Full Text Available The reversible acetylation of lysine residues is catalyzed by the antagonistic action of lysine acetyltransferases and deacetylases, which can be considered as master regulators of their substrate proteins. Lysine deacetylases, historically referred to as histone deacetylases, have profound functions in regulating stress defenses and development in plants. Lysine acetylation of the N-terminal histone tails promotes gene transcription and decondensation of chromatin, rendering the DNA more accessible to the transcription machinery. In plants, the classical lysine deacetylases from the RPD3/HDA1-family have thus far mainly been studied in the context of their deacetylating activities on histones, and their versatility in molecular activities is still largely unexplored. Here we discuss the potential impact of lysine acetylation on the recently identified nuclear substrate proteins of lysine deacetylases from the Arabidopsis RPD3/HDA1-family. Among the deacetylase substrate proteins, many interesting candidates involved in nuclear protein import, transcriptional regulation, and chromatin remodeling have been identified. These candidate proteins represent key starting points for unraveling new molecular functions of the Arabidopsis lysine deacetylases. Site-directed engineering of lysine acetylation sites on these target proteins might even represent a new approach for optimizing plant growth under climate change conditions.

  7. Minoxidil specifically decreases the expression of lysine hydroxylase in cultured human skin fibroblasts.

    Science.gov (United States)

    Hautala, T; Heikkinen, J; Kivirikko, K I; Myllylä, R

    1992-01-01

    The levels of lysine hydroxylase protein and the levels of the mRNAs for lysine hydroxylase and the alpha- and beta-subunits of proline 4-hydroxylase were measured in cultured human skin fibroblasts treated with 1 mM-minoxidil. The data demonstrate that minoxidil decreases the amount of lysine hydroxylase protein, this being due to a decrease in the level of lysine hydroxylase mRNA. The effect of minoxidil appears to be highly specific, as no changes were observed in the amounts of mRNAs for the alpha- and beta-subunits of proline 4-hydroxylase. Images Fig. 1. Fig. 2. Fig. 3. PMID:1314568

  8. Evolving minimum standards in responsible international sperm donor offspring quota

    NARCIS (Netherlands)

    Janssens, Pim M. W.; Thorn, Petra; Castilla, Jose A.; Frith, Lucy; Crawshaw, Marilyn; Mochtar, Monique; Bjorndahl, Lars; Kvist, Ulrik; Kirkman-Brown, Jackson C.

    2015-01-01

    An international working group was established with the aim of making recommendations on the number of offspring for a sperm donor that should be allowable in cases of international use of his sperm. Considerations from genetic, psychosocial, operational and ethical points of view were debated. For

  9. Sperm DNA damage in relation to lipid peroxidation following ...

    African Journals Online (AJOL)

    This study investigated the relationships between lipid peroxidation (LPO) and sperm DNA damage following freezing-thawing of boar semen in different extenders. The comet assay was used to measure the extent of sperm DNA damage in a cryoprotectant-free extender or in cryoprotectant-based extenders after single ...

  10. Reproductive tract modifications of the boar sperm surface

    NARCIS (Netherlands)

    Gadella, Bart M|info:eu-repo/dai/nl/115389873

    2017-01-01

    The sperm cell has a unique, polarized and segregated surface that is modified extensively by the changing environments in both the male and the female reproductive tracts. The sperm cannot refresh its surface, as protein translation and membrane recycling by intracellular vesicular transport have

  11. Sperm use economy of honeybee (Apis mellifera) queens

    DEFF Research Database (Denmark)

    Baer, Boris; Collins, Jason; Maalaps, Kristiina

    2016-01-01

    the fecundity and longevity of queens and therefore colony fitness. We quantified the number of sperm that honeybee (Apis mellifera) queens use to fertilize eggs. We examined sperm use in naturally mated queens of different ages and in queens artificially inseminated with different volumes of semen. We found...

  12. Modifications in sperm quality of Wister Albino Rats by Ethanol ...

    African Journals Online (AJOL)

    The effect of a well known herb, Phyllanthus amarus on the sperm characteristics in male albino rats was studied. This was an investigation of the age-long claim by the locales in the rural communities in the southern states of Nigeria where this plant is consumed religiously that it affects sperm quality, hence sexual potency ...

  13. Microscopic analysis of MTT stained boar sperm cells

    African Journals Online (AJOL)

    tulyasys

    2015-06-08

    Jun 8, 2015 ... Microscopic analysis of MTT stained boar sperm cells. B.M. van den Berg*. Barex Biochemical Products, Seb. Centenweg 45, 1602ML Enkhuizen, The Netherlands. Abstract. The ability of sperm cells to develop colored formazan by reduction of MTT was used earlier to develop a spectrophotometric.

  14. Modifications in sperm quality of Wister Albino Rats by Ethanol ...

    African Journals Online (AJOL)

    Prof. Ogunji

    Abstract. The effect of a well known herb, Phyllanthus amarus on the sperm characteristics in male albino rats was studied. This was an investigation of the age-long claim by the locales in the rural communities in the southern states of Nigeria where this plant is consumed religiously that it affects sperm quality, hence ...

  15. A Study of Anti-Sperm Antibodies among Infertile Subjects ...

    African Journals Online (AJOL)

    Background: Anti-sperm antibodies (ASA) can interfere with sperm functions and fertility and may be found in the blood, lymph or local secretions such as seminal and cervico-vaginal fluids in both men and women. Objective: to evaluate the contribution of ASA to infertility in male and female subjects investigated for ...

  16. Cephalopods in the diet of sperm whales caught commercially off ...

    African Journals Online (AJOL)

    A collection from stomach contents of 30 sperm whales Physeter catodon comprised a total of 46 cephalopods belonging to six families. Nine species were identified, including Ommastrephes bartramii, which is recorded for the first time in the diet of sperm whales caught off South Africa, and Todarodes filippovae, which has ...

  17. The effect of sperm morphology and testicular spermatozoa ...

    African Journals Online (AJOL)

    Objective. To determine the correlation between sperm morphology groups (strict criteria) and testicular spermatozoa, and day 2 and 3 embryo quality in intracytoplasmic sperm injection (ICSI) and in vitro fertilisation (IVF) cases. Methods. A retrospective study was done of 2 402 IVF and ICSI-fertilised embryos classified as ...

  18. Evidence for decreasing sperm count in African population from ...

    African Journals Online (AJOL)

    Purpose: This meta-analysis, following our previous reports those documented an overall 57% diminution in mean sperm concentration around the globe over past 35 years and 32.5% decline in past 50 years in European population, attempts to report the declining trend of sperm concentrations in African population ...

  19. in human sperm motility and level of calcium and magnesium

    African Journals Online (AJOL)

    J. Valsa

    2015-11-06

    Nov 6, 2015 ... Abstract A detailed sperm motility study for 24 h after collection was done. The level of calcium and magnesium in seminal plasma during this period was also seen to understand the role of these electrolytes on sperm motility. Good care was taken in selection of subjects (young and healthy), collection and ...

  20. Maintenance of sperm variation in a highly promiscuous wild bird.

    Science.gov (United States)

    Calhim, Sara; Double, Michael C; Margraf, Nicolas; Birkhead, Tim R; Cockburn, Andrew

    2011-01-01

    Postcopulatory sexual selection is an important force in the evolution of reproductive traits, including sperm morphology. In birds, sperm morphology is known to be highly heritable and largely condition-independent. Theory predicts, and recent comparative work corroborates, that strong selection in such traits reduces intraspecific phenotypic variation. Here we show that some variation can be maintained despite extreme promiscuity, as a result of opposing, copulation-role-specific selection forces. After controlling for known correlates of siring success in the superb fairy-wren (Malurus cyaneus), we found that (a) lifetime extra-pair paternity success was associated with sperm with a shorter flagellum and relatively large head, and (b) males whose sperm had a longer flagellum and a relatively smaller head achieved higher within-pair paternity. In this species extrapair copulations occur in the same morning, but preceding, pair copulations during a female's fertile period, suggesting that shorter and relatively larger-headed sperm are most successful in securing storage (defense), whereas the opposite phenotype might be better at outcompeting stored sperm (offense). Furthermore, since cuckolding ability is a major contributor to differential male reproductive output, stronger selection on defense sperm competition traits might explain the short sperm of malurids relative to other promiscuous passerines.

  1. Maintenance of sperm variation in a highly promiscuous wild bird.

    Directory of Open Access Journals (Sweden)

    Sara Calhim

    Full Text Available Postcopulatory sexual selection is an important force in the evolution of reproductive traits, including sperm morphology. In birds, sperm morphology is known to be highly heritable and largely condition-independent. Theory predicts, and recent comparative work corroborates, that strong selection in such traits reduces intraspecific phenotypic variation. Here we show that some variation can be maintained despite extreme promiscuity, as a result of opposing, copulation-role-specific selection forces. After controlling for known correlates of siring success in the superb fairy-wren (Malurus cyaneus, we found that (a lifetime extra-pair paternity success was associated with sperm with a shorter flagellum and relatively large head, and (b males whose sperm had a longer flagellum and a relatively smaller head achieved higher within-pair paternity. In this species extrapair copulations occur in the same morning, but preceding, pair copulations during a female's fertile period, suggesting that shorter and relatively larger-headed sperm are most successful in securing storage (defense, whereas the opposite phenotype might be better at outcompeting stored sperm (offense. Furthermore, since cuckolding ability is a major contributor to differential male reproductive output, stronger selection on defense sperm competition traits might explain the short sperm of malurids relative to other promiscuous passerines.

  2. Heritability of sperm length in the bumblebee Bombus terrestris

    DEFF Research Database (Denmark)

    Baer, Boris; de Jong, Gerdien; Schmid-Hempel, Regula

    2006-01-01

    Sperm length is highly variable, both between and within species, but the evolutionary significance of this variation is poorly understood. Sexual selection on sperm length requires a significant additive genetic variance, but few studies have actually measured this. Here we present the first est...

  3. Evidence for decreasing sperm count in African population from ...

    African Journals Online (AJOL)

    Abstract. Purpose: This meta-analysis, following our previous reports those documented an overall 57% diminution in mean sperm con- centration around the globe over past 35 years and 32.5% decline in past 50 years in European population, attempts to report the declining trend of sperm concentrations in African ...

  4. Diisopropyl fluorophosphate labeling of sperm-associated proteinases

    International Nuclear Information System (INIS)

    Odem, R.R.; Willand, J.L.; Polakoski, K.L.

    1990-01-01

    Proteinase inhibitors have been shown to be capable of preventing various aspects of fertilization. Diisopropyl fluorophosphate (DFP) is an irreversible inhibitor of trypsin-like enzymes that is commercially available in a radiolabeled form. The experiments described herein were designed to determine if DFP would prevent sperm function in live, motile sperm and to identify the sperm proteins bound with DFP. DFP at 5 mM concentrations had no observable effect on sperm motility, but inhibited the penetration of zona-free hamster ova by human sperm (5.5%) compared to controls (33.5%). Acid extracts of motile sperm that had been incubated with radiolabeled DFP and collected by the swim-up procedure demonstrated the presence of radiolabeled DFP, and the autoradiography of the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gels of these extracts localized the uptake of radiolabeled DFP to proteins in the molecular weight region of the proacrosin-acrosin system. Acid-extracted proteinases from semen samples incubated with DFP demonstrated a concentration-dependent inhibition of both esterolytic hydrolysis of benzoyl-arginine ethyl ester on spectrophotometric analysis and proteolytic activity on gelatin SDS-PAGE zymography. DFP-labeled proteins were precipitated by highly specific antibodies to proacrosin. These results demonstrated that DFP is capable of inhibiting sperm function, and that it associates with the proacrosin-acrosin system in live motile sperm

  5. Diisopropyl fluorophosphate labeling of sperm-associated proteinases

    Energy Technology Data Exchange (ETDEWEB)

    Odem, R.R.; Willand, J.L.; Polakoski, K.L. (Washington Univ. School of Medicine, St. Louis, MO (USA))

    1990-02-01

    Proteinase inhibitors have been shown to be capable of preventing various aspects of fertilization. Diisopropyl fluorophosphate (DFP) is an irreversible inhibitor of trypsin-like enzymes that is commercially available in a radiolabeled form. The experiments described herein were designed to determine if DFP would prevent sperm function in live, motile sperm and to identify the sperm proteins bound with DFP. DFP at 5 mM concentrations had no observable effect on sperm motility, but inhibited the penetration of zona-free hamster ova by human sperm (5.5%) compared to controls (33.5%). Acid extracts of motile sperm that had been incubated with radiolabeled DFP and collected by the swim-up procedure demonstrated the presence of radiolabeled DFP, and the autoradiography of the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gels of these extracts localized the uptake of radiolabeled DFP to proteins in the molecular weight region of the proacrosin-acrosin system. Acid-extracted proteinases from semen samples incubated with DFP demonstrated a concentration-dependent inhibition of both esterolytic hydrolysis of benzoyl-arginine ethyl ester on spectrophotometric analysis and proteolytic activity on gelatin SDS-PAGE zymography. DFP-labeled proteins were precipitated by highly specific antibodies to proacrosin. These results demonstrated that DFP is capable of inhibiting sperm function, and that it associates with the proacrosin-acrosin system in live motile sperm.

  6. A sperm competition model for the European bitterling (Rhodeus amarus)

    Czech Academy of Sciences Publication Activity Database

    Smith, Carl; Reichard, Martin

    2013-01-01

    Roč. 150, č. 14 (2013), s. 1709-1730 ISSN 0005-7959 R&D Projects: GA ČR GA206/09/1163 Institutional support: RVO:68081766 Keywords : Acheilognathinae * alternative mating tactics * mating system * sneaking * sperm competition * sperm motility Subject RIV: EG - Zoology Impact factor: 1.401, year: 2013

  7. Sperm banking for male cancer patients: social and semen profiles

    Directory of Open Access Journals (Sweden)

    Tatiana C.S. Bonetti

    2009-04-01

    Full Text Available PURPOSE: Report the characteristics of cryopreserved semen from a cohort of male cancer patients, attitudes towards cryopreservation and outcomes of semen samples based on a 12-year cryopreservation program. MATERIAL AND METHODS: Data from 98 male cancer patients whose sperm samples were banked were evaluated. Demographic parameters, semen characteristics, destination of sperm banked samples and questionnaires answered by the patients regarding cryopreservation time were evaluated. RESULTS: The cancer diagnoses were testicle (56.1%, prostate (15.3%, Hodgkin’s lymphomas (9.2%, non-Hodgkin’s lymphomas (7.1%, leukemia (3.1% and other malignancies (9.2%. The patients with testicular cancer presented lower sperm concentration (p < 0.001; however, there were no differences with the percentage of normozoospermic patients among cancer type groups (p = 0.185. A shorter time between cancer diagnosis and sperm banking was observed for testicular and prostate cancer patients (p < 0.001. Most of the patients (89.5% favored sperm banking as a fertility preservation method. CONCLUSIONS: Although less than 20% of banked sperm samples were disposed of, the majority of patients related sperm banking with safe for fertility preservation. Our results show that all male cancer patients of reproductive age facing cancer treatment could be offered sperm banking.

  8. CatSper channel, sperm function and male fertility.

    Science.gov (United States)

    Singh, Akhand Pratap; Rajender, Singh

    2015-01-01

    A number of physiological events, such as sperm hyperactivation, chemotaxis towards the egg, capacitation and acrosome reaction, are triggered by activation of sperm ion channels in response to a diverse range of chemical cues. Cation channel of sperm (CatSper), a sperm-specific ion channel, is unique in orchestrating the events for fertilization, and seems to be exclusively evolved for sperm function and male fertility. CatSper acts as a polymodal, chemosensory calcium channel and plays a vital role in the regulation of sperm hyperactivation. CatSper knockout models and application of patch clamp recordings have shown that it is indispensable for male fertility, and mutations and deletions in CatSper gene(s) may lead to infertility. In fact, mutations in CatSper1 and 2 have been identified in infertile individuals; however, CatSper3 and 4 have not been explored. Restricted localization and expression of CatSper in sperm offer an added advantage to developing gamete-based safe non-hormonal contraceptives. This review concisely covers identification, structure, function, and mechanism of action of CatSper channels. The functional importance of this complex ion channel in sperm motility and male fertility is highlighted for further research on male fertility, infertility, and contraception. Copyright © 2014 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.

  9. Enhancement of Sexual Libido and Sperm Production in rabbits ...

    African Journals Online (AJOL)

    Enhancement of Sexual Libido and Sperm Production in rabbits using Clomiphene citrate (Clomid®) ... The results obtained in Trial II showed that sperm concentration was higher (P<0.05) in animals treated with 6.25mg of the drug than their counterparts treated with 12.50mg of the drug. The Chinchilla bucks were of higher ...

  10. Human sperm swimming in a high viscosity mucus analogue.

    Science.gov (United States)

    Ishimoto, Kenta; Gadêlha, Hermes; Gaffney, Eamonn A; Smith, David J; Kirkman-Brown, Jackson

    2018-02-17

    Remarkably, mammalian sperm maintain a substantive proportion of their progressive swimming speed within highly viscous fluids, including those of the female reproductive tract. Here, we analyse the digital microscopy of a human sperm swimming in a highly viscous, weakly elastic mucus analogue. We exploit principal component analysis to simplify its flagellar beat pattern, from which boundary element calculations are used to determine the time-dependent flow field around the sperm cell. The sperm flow field is further approximated in terms of regularized point forces, and estimates of the mechanical power consumption are determined, for comparison with analogous low viscosity media studies. This highlights extensive differences in the structure of the flows surrounding human sperm in different media, indicating how the cell-cell and cell-boundary hydrodynamic interactions significantly differ with the physical microenvironment. The regularized point force decomposition also provides cell-level information that may ultimately be incorporated into sperm population models. We further observe indications that the core feature in explaining the effectiveness of sperm swimming in high viscosity media is the loss of cell yawing, which is related with a greater density of regularized point force singularities along the axis of symmetry of the flagellar beat to represent the flow field. In turn this implicates a reduction of the wavelength of the distal beat pattern - and hence dynamical wavelength selection of the flagellar beat - as the dominant feature governing the effectiveness of sperm swimming in highly viscous media. Copyright © 2018. Published by Elsevier Ltd.

  11. ROLE OF APOPTOSIS IN THE EVALUATION OF SPERM QUALITY ...

    African Journals Online (AJOL)

    Results In the fertile men (control group), none of the sperms was positive for DNA strand breaks, while in the oligozoospermic patients 0.9 0.7% of the sperms were positive for DNA strand breaks. This result was statistically highly ... dans les traitements avancés de fertilisation. African Journal of Urology Vol.8(1) 2002: 6-12 ...

  12. Sperm competition-induced plasticity in the speed of spermatogenesis.

    Science.gov (United States)

    Giannakara, Athina; Schärer, Lukas; Ramm, Steven A

    2016-03-08

    Sperm competition between rival ejaculates over the fertilization of ova typically selects for the production of large numbers of sperm. An obvious way to increase sperm production is to increase testis size, and most empirical work has focussed on this parameter. Adaptive plasticity in sperm production rate could also arise due to variation in the speed with which each spermatozoon is produced, but whether animals can respond to relevant environmental conditions by modulating the kinetics of spermatogenesis in this way has not been experimentally investigated. Here we demonstrate that the simultaneously hermaphroditic flatworm Macrostomum lignano exhibits substantial plasticity in the speed of spermatogenesis, depending on the social context: worms raised under higher levels of sperm competition produce sperm faster. Our findings overturn the prevailing view that the speed of spermatogenesis is a static property of a genotype, and demonstrate the profound impact that social environmental conditions can exert upon a key developmental process. We thus identify, to our knowledge, a novel mechanism through which sperm production rate is maximised under sperm competition.

  13. Alkaline phosphatase in boar sperm function.

    Science.gov (United States)

    Bucci, D; Isani, G; Giaretta, E; Spinaci, M; Tamanini, C; Ferlizza, E; Galeati, G

    2014-01-01

    Alkaline phosphatase (AP) catalyses the detachment of phosphate residues from different substrates. Its activity has been demonstrated in seminal plasma and spermatozoa from porcine and other mammalian species; anyway, the role of AP in male reproduction has not been clarified yet and the aim of this study was to determine AP function in boar sperm capacitation and in vitro fertilization (IVF). AP activity was assayed in seminal plasma and in uncapacitated and in vitro capacitated (IVC) spermatozoa; in addition, capacitation was studied in presence of different doses of AP (1.2 and 2.5 IU/mL). The effect of different doses of AP (1.2 and 2.5 IU/mL) on several sperm parameters after IVC (viability, acrosome integrity with FITC-PSA, capacitation status with CTC staining, tyrosine phosphorylation) and on fertilizing ability during IVF were also evaluated. High AP activity was detected in seminal plasma, in particular in sperm-rich fraction; a lower activity was detected in uncapacitated spermatozoa while a significant decrease was evidenced after IVC. Viability was not changed by AP supplementation of the capacitating medium, whereas acrosome integrity and capacitation status were significantly affected by 1.2 and 2.5 doses, with a dose-dependent decrease in acrosome-reacted cells as well as in CTC B pattern displaying cells. As for sperm head protein phosphorylation, a decrease in relative fluorescence was detected in AP 2.5 group, if compared with capacitated one. After IVF, a dose-dependent decrease in penetrated oocytes was recorded, with an increase in monospermic zygote rate. In conclusion, we demonstrated that AP activity decreases under capacitating condition and that addition of AP to spermatozoa during capacitation results in a depression of the capacitating process and IVF. We can infer that AP plays a role in keeping spermatozoa quiescent until they are ejaculated and in modulating the acquisition of the fertilizing ability. © 2013 American Society of

  14. Methods useful for evaluation of human sperm

    Czech Academy of Sciences Publication Activity Database

    Kubátová, Alena; Čapková, Jana; Pěknicová, Jana

    2012-01-01

    Roč. 67, Issue Supplement s1 (2012), s. 27-27 ISSN 1046-7408. [13th International Symposium for Immunology of reproduction "From the roots to the tops of Reproductive Immunology". 22.06.2012-24.06.2012, Varna] R&D Projects: GA ČR(CZ) GA523/09/1793; GA ČR(CZ) GAP503/12/1834 Institutional research plan: CEZ:AV0Z50520701 Keywords : human sperm * immunofluorescence test * human seminal plasma proteins * flow cytometry Subject RIV: EC - Immunology

  15. Sperm DNA fragmentation and oxidation are independent of malondialdheyde

    Science.gov (United States)

    2011-01-01

    Background There is clinical evidence to show that sperm DNA damage could be a marker of sperm quality and extensive data exist on the relationship between DNA damage and male fertility status. Detecting such damage in sperm could provide new elements besides semen parameters in diagnosing male infertility. We aimed to assess sperm DNA fragmentation and oxidation and to study the association between these two markers, routine semen parameters and malondialdehyde formation. Methods Semen samples from 55 men attending the Histology-Embryology Laboratory of Sfax Faculty of Medicine, Tunisia, for semen investigations were analysed for sperm DNA fragmentation and oxidation using flow cytometry. The Sperm was also assessed spectrophotometrically for malondialdehyde formation. Results Within the studied group, 21 patients were nonasthenozoospermic (sperm motility ≥ 50%) and 34 patients were considered asthenozoospermic (sperm motility < 50%). A positive correlation was found between sperm DNA fragmentation and oxidation (p = 0.01; r = 0.33). We also found a negative correlation between sperm DNA fragmentation and some sperm parameters: total motility (p = 0.001; r = -0.43), rapid progressive motility (type a motility) (p = 0.04; r = -0.27), slow progressive motility (type b motility) (p = 0.03; r = -0.28), and vitality (p < 0.001; r = -0.65). Sperm DNA fragmentation was positively correlated with coiled tail (p = 0.01; r = 0.34). The two parameters that were found to be correlated with oxidative DNA damage were leucocytes concentrations (p = 0.01; r = 0.38) and broken neck (p = 0.02; r = 0.29). Sperm MDA levels were negatively correlated with sperm concentration (p < 0.001; r = -0.57), total motility (p = 0.01; r = -0.35) and type a motility (p = 0.03; r = -0.32); but not correlated with DNA fragmentation and DNA oxidation. Conclusions Our results support the evidence that oxidative stress plays a key role in inducing DNA damage; but nuclear alterations and

  16. Sperm aggregation by water extracts from two Bursera species.

    Science.gov (United States)

    Serrano, H; Garcia-Suarez, M D

    2001-01-01

    Some plants have more than the common utility value, as is the case of some members of the Bursera species such as the Mexican copal, a plant used for worship. Water extracts of several plants have vaginal contraceptive properties. The authors evaluated the sperm agglutinating activity of two Bursera species on human and boar sperm. Extracts from stems and leaves were obtained. Capacitated sperm samples were used in all cases. There were different agglutinating capacities, which were not observed in the vehicle-only samples. The most frequent sperm agglutination response was that involving the heads. Agglutinating activity was higher from stem- than leaf-derived extracts. The results indicate that proteins present in the extracts are responsible for the aggregation of sperm heads.

  17. Shape and shear guide sperm cells spiraling upstream

    Science.gov (United States)

    Kantsler, Vasily; Dunkel, Jorn; Goldstein, Raymond E.

    2014-11-01

    A major puzzle in biology is how mammalian sperm determine and maintain the correct swimming direction during the various phases of the sexual reproduction process. Currently debated mechanisms for sperm long range travel vary from peristaltic pumping to temperature sensing (thermotaxis) and direct response to fluid flow (rheotaxis), but little is known quantitatively about their relative importance. Here, we report the first quantitative experimental study of mammalian sperm rheotaxis. Using microfluidic devices, we investigate systematically the swimming behavior of human and bull sperm over a wide range of physiologically relevant shear rates and viscosities. Our measurements show that the interplay of fluid shear, steric surface-interactions and chirality of the flagellar beat leads to a stable upstream spiraling motion of sperm cells, thus providing a generic and robust rectification mechanism to support mammalian fertilization. To rationalize these findings, we identify a minimal mathematical model that is capable of describing quantitatively the experimental observations.

  18. Testicular Damage following Testicular Sperm Retrieval: A Ram Model Study

    Directory of Open Access Journals (Sweden)

    Jens Fedder

    2017-01-01

    Full Text Available The aim of this study was to evaluate the possible development of histological abnormalities such as fibrosis and microcalcifications after sperm retrieval in a ram model. Fourteen testicles in nine rams were exposed to open biopsy, multiple TESAs, or TESE, and the remaining four testicles were left unoperated on as controls. Three months after sperm retrieval, the testicles were removed, fixed, and cut into 1/2 cm thick slices and systematically put onto a glass plate exposing macroscopic abnormalities. Tissue from abnormal areas was cut into 3 μm sections and stained for histological evaluation. Pathological abnormalities were observed in testicles exposed to sperm retrieval (≥11 of 14 compared to 0 of 4 control testicles. Testicular damage was found independently of the kind of intervention used. Therefore, cryopreservation of excess sperm should be considered while retrieving sperm.

  19. Bending the law and crossing borders choosing Danish sperm

    DEFF Research Database (Denmark)

    Willum Adrian, Stine

    In recent years, Denmark has become a European destination for couples and single women wanting sperm donation. One reason is that the current regulation is liberal in the sense that it enables single women and lesbians to be treated. Since neither private sperm banks nor clinics run by midwifes...... have to apply to the law, it is even possible to choose between anonymous and non-anonymous donors. In this presentation, I focus on how the fertility travellers seeking sperm donation in Denmark, and the employees at Danish fertility clinics and sperm banks, negotiate the ethical implications of using...... sperm donation. Inspired by Karen Barad’s theory agential realism, I will analyze how ethics is material-discoursively negotiated in practice. The presentation is based on an ethnographic fieldwork carried out at fertility clinics in Denmark. It includes observations from the clinics, and interviews...

  20. Human sperm steer with second harmonics of the flagellar beat.

    Science.gov (United States)

    Saggiorato, Guglielmo; Alvarez, Luis; Jikeli, Jan F; Kaupp, U Benjamin; Gompper, Gerhard; Elgeti, Jens

    2017-11-10

    Sperm are propelled by bending waves traveling along their flagellum. For steering in gradients of sensory cues, sperm adjust the flagellar waveform. Symmetric and asymmetric waveforms result in straight and curved swimming paths, respectively. Two mechanisms causing spatially asymmetric waveforms have been proposed: an average flagellar curvature and buckling. We image flagella of human sperm tethered with the head to a surface. The waveform is characterized by a fundamental beat frequency and its second harmonic. The superposition of harmonics breaks the beat symmetry temporally rather than spatially. As a result, sperm rotate around the tethering point. The rotation velocity is determined by the second-harmonic amplitude and phase. Stimulation with the female sex hormone progesterone enhances the second-harmonic contribution and, thereby, modulates sperm rotation. Higher beat frequency components exist in other flagellated cells; therefore, this steering mechanism might be widespread and could inspire the design of synthetic microswimmers.