scholarly journals Cryopreservation of spermatozoa of indigenous stinging catfish, Heteropneustes fossilis (Bloch) for ex-situ conservation

2020 ◽  
Vol 32 (1) ◽  
pp. 11-22
Author(s):  
MD. RAFIQUL ISLAM SARDER ◽  
MD. ABUL KALAM AZAD ◽  
K.M. SHAHRIAR NAZRUL ◽  
MOHAMMAD RASHED
Keyword(s):  
Ex Situ Conservation ◽  
Egg Yolk ◽  
Ringer Solution ◽  
The Other ◽  
Ex Situ ◽  
Nacl Solution ◽  
Heteropneustes Fossilis ◽  
Significant Difference ◽  
Cryopreserved Sperm ◽  
Cryopreservation Protocol

An experiment was conducted to develop cryopreservation protocol for spermatozoa of stinging catfish, Heteropneustes fossilis and to use the cryopreserved sperm in its breeding trials. The activation of sperm motility at various concentrations of NaCl solution was tested and complete activation and inhibition of sperm were obtained at 0.4% and 0.9% to 1% NaCl solution respectively. In toxicity test, sperms were incubated with DMSO, methanol and ethanol at 5, 10, and 15% concentrations where DMSO and methanol produced better motility at 5 and 10% concentration with Alsever’s solution and egg-yolk citrate at 5 and- 10 min incubation period. Three extenders- Alsever’s solution, egg-yolk citrate and Ginsburg Fish Ringer solution and three cryoprotectants- DMSO, methanol and ethanol were used for cryopreservation of sperm, and among the diluents, Alsever’s solution with 10% DMSO showed best performance producing 77.50±3.22% post-thaw motility. On the other hand, egg-yolk citrate and Ginsburg Fish Ringer solution along with 10% DMSO produced 63.75±2.39% and 62.50±3.22% post-thaw motility, respectively. Sperm preserved with Alsever’s solution plus 10% DMSO produced 52.5±3.34% and 38.0±2.39%fertilization and hatching, and those preserved with Alsever’s solution plus 10% methanol produced 46.9±3.11% and 32.7±2.70% fertilization and hatching respectively. The fry produced using cryopreserved and fresh sperm grew well and no significant difference (p>0.05) was found between two groups.

Development of sperm cryopreservation protocol of endangered spiny eel, Mastacembelus armatus (Lacepede 1800) for ex-situ conservation

Cryobiology ◽  
2016 ◽  
Vol 73 (3) ◽  
pp. 316-323 ◽  
Author(s):  
M. Mahfujur Rahman ◽  
M. Rahmat Ali ◽  
M. Rafiqul Islam Sarder ◽  
M. Fazlul Awal Mollah ◽  
Najmus Sakib Khan
Keyword(s):  
Ex Situ Conservation ◽  
Ex Situ ◽  
Sperm Cryopreservation ◽  
Mastacembelus Armatus ◽  
Cryopreservation Protocol

scholarly journals DNA fragmentation and membrane damage of bocachico Prochilodus magdalenae (Ostariophysi: Prochilodontidae) sperm following cryopreservation with dimethylsulfoxide and glucose

2012 ◽  
Vol 10 (3) ◽  
pp. 577-586 ◽  
Author(s):  
José Gregorio Martínez ◽  
Víctor Atencio García ◽  
Sandra Pardo Carrasco
Keyword(s):  
Dna Fragmentation ◽  
Ex Situ Conservation ◽  
Sperm Quality ◽  
Membrane Damage ◽  
Egg Yolk ◽  
Ex Situ ◽  
Important Species ◽  
Chicken Egg ◽  
Nitrogen Vapor ◽  
Chicken Egg Yolk

The endangered bocachico Prochilodus magdalenae is a native freshwater fish of Colombia, the most captured species locally and one of the most important species for ex-situ conservation (germplasm banks). The aim of this study was to examine the effect of three concentrations of Dimethylsulfoxide (DMSO) (5%, 10%, 15%) and three of glucose (305, 333, 361 mM) in the extender on spermatic DNA fragmentation (F-DNA) (by Halomax®, Chromatin dispersion) and membrane damage (D-Me) (by eosin-nigrosin staining). After assessment of sperm quality by computer analysis of motility, one part of semen from males was diluted separately with three parts of extender and filled into 0.5 ml straws. Freezing was carried out in liquid nitrogen vapor dry shipper for 30 minutes and thawed at 60ºC for 8 seconds in a water bath and evaluated for the percentage of cells found with F-DNA and D-Me. The results demonstrated that cryopreservation causes greater F-DNA (13.62 ± 1.6% to 28.91 ± 3.25) and D-Me (24.27 ± 1.1% to 58.33 ± 2.81%) when compared with pre-freezing semen (PFS) (6.71 ± 1.54% and 2.34 ± 0.5%, respectively for F-DNA and D-Me). A significant interaction was found between DMSO and glucose concentration in this experiment. Use of extender: 10% DMSO + 305 mM glucose + 12% chicken egg yolk and, 10% DMSO + 333 mM glucose + 12% chicken egg yolk, allow for lower F-DNA and D-Me during cryopreservation of bocachico semen. A high correlation between F-DNA and D-Me was found (r = 0.771).

Effects of Different Molting Procedures on Incidence of Salmonella Infection in Flocks of Naturally Contaminated Laying Hens in a Commercial Egg-Producing Farm by Detection of Yolk Antibodies to Salmonella in Eggs

2006 ◽  
Vol 69 (12) ◽  
pp. 2883-2888 ◽  
Author(s):  
TOSHIYUKI MURASE ◽  
KAORI CHIBA ◽  
TOMOKO SATO ◽  
KOICHI OTSUKI ◽  
PETER S. HOLT
Keyword(s):  
Wheat Bran ◽  
Egg Production ◽  
Egg Yolk ◽  
The Other ◽  
Feed Withdrawal ◽  
Two Samples ◽  
Significant Difference ◽  
Before And After ◽  
Specific Pathogen ◽  
Egg Yolks

Indirect enzyme-linked immunosorbent assays (ELISAs) have been applied to detect immunoglobulin Y antibodies to different serotypes of Salmonella in the yolks of chicken eggs with heat-extracted antigens of Salmonella enterica serotypes Agona (SA), Cerro (SC), Enteritidis (SE), Montevideo (SM), and Putten (SP). The egg yolk samples examined were classified as positive if their ELISA absorbance values exceeded the value for eggs from specific-pathogen-free flocks by more than two standard deviations. Of 30 egg yolk samples from three flocks vaccinated with a killed SE vaccine, 29 were antibody positive by the ELISA assay for the SE antigen. Four to 29 of the 29 yolk samples showed positive results for the other serovars, although the absorbance values for SE were higher than those obtained for the other serotypes in each of the yolk samples. All 30 yolks from three flocks that were not administered any SE vaccines were found to be antibody negative for SE, and two samples were determined to be positive for SC. Thirty-nine or 40 eggs were obtained from each of four layer flocks in a commercial egg production farm where the laying houses were naturally contaminated with SA, SC, SM, SP, Salmonella serovar Infantis (SI), and untypeable strains. The ELISA absorbance values for SM in the egg yolks obtained from the two flocks molted through feed withdrawal when the birds restarted laying were significantly (P < 0.05) higher than those observed in the yolks obtained before the molt. In egg yolks from the two other flocks that were molted through a wheat bran diet, there was no significant difference between the absorbance values before and after the molt. The observations in the present study provide further evidence to suggest that a molt initiated through the administration of a wheat bran diet can reduce the risk for Salmonella problems in a commercial egg-producing setting.

scholarly journals KAJIAN ELONGASI PADA TANAMAN IN VITRO GAHARU (Aquilaria beccariana van Tiegh)

2015 ◽  
Vol 2 (2) ◽  
pp. 65
Author(s):  
Yusuf Sigit Ahmad Fauzan ◽  
Edhi Sandra ◽  
Daru Mulyono
Keyword(s):  
In Vitro Propagation ◽  
Ex Situ Conservation ◽  
Shoot Elongation ◽  
The Other ◽  
Ex Situ ◽  
Average Height ◽  
Best Response ◽  
Speed Up ◽  
Number Of Segments

The population density of natural agarwood (Aquilaria beccariana) in Indonesia decreased to less than one tree per hectare. Efforts have been carried out on ex situ conservation of agarwood despite facing many obstacles. In vitro propagation is one alternative to speed up the recovery of natural agarwood populations. The purpose of this study was to obtain optimal elongation media for in vitro culture with addition of auxin and cytokinin, namely IBA, BAP and kinetin. The results showed that the best auxin-cytokinin combination was IBA 0.1 mg/L and BAP 0.05 mg/L. This combination increased the height and number of segments of A. beccariana with an average height of 1.64 cm and average number of sections of 6.40. It is suggested that this combination of IBA and BAP was the most effective compared to the other treatments. In addition, the combination of IBA 0 mg/L and BAP 0.03 mg/L gave rise to the best response to increase the number of shoots with an average of 1.91 shoots.Keywords: Aquilaria beccariana, shoot, elongation, auxin, cytokinin ABSTRAKKepadatan populasi gaharu (Aquilaria beccariana) alam di Indonesia kurang dari satu pohon per hektar. Upaya pelestarian gaharu ex situ telah banyak dilakukan tetapi masih banyak kendala. Perbanyakan gaharu in vitro merupakan salah satu cara alternatif untuk mempercepat pemulihan populasi gaharu alam. Tujuan penelitian ini adalah untuk memperoleh media elongasi yang optimal pada kultur in vitro gaharu dengan penambahan kombinasi zat pengatur tumbuh auksin dan sitokinin. Pada penelitian ini digunakan auksin IBA, serta sitokinin BAP dan Kinetin. Hasil penelitian elongasi diperoleh kombinasi auksin dan sitokinin terbaik yaitu, IBA 0,1 mg/L dan BAP 0,05 mg/L. Kombinasi ini meningkatkan tinggi dan jumlah ruas Aquilaria beccariana dengan tinggi rata-rata sebesar 1,64 cm dan jumlah ruas rata-rata sebesar 6,40 ruas. Pada kombinasi dan taraf ini diduga mekanisme kerja IBA dan BAP paling efektif dibanding perlakuan yang lain. Sedangkan kombinasi IBA 0 mg/L dan BAP 0,03 mg/L memberikan respon terbaik terhadap peningkatan jumlah tunas dengan rata-rata sebanyak 1,91 tunas.Kata Kunci: Aquilaria beccariana, tunas, elongasi, auksin, sitokinin

scholarly journals Ultra-low temperature conservation of Brazilian Pine embryogenic cultures

2014 ◽  
Vol 86 (4) ◽  
pp. 2057-2064 ◽  
Author(s):  
GRASIELA DEMARCHI ◽  
VALDIR M. STEFENON ◽  
NEUSA STEINER ◽  
FELIPE N. VIEIRA ◽  
LIRIO L. DAL VESCO ◽  
...  
Keyword(s):  
Low Temperature ◽  
Cell Viability ◽  
Biochemical Composition ◽  
Ex Situ Conservation ◽  
Ex Situ ◽  
Embryogenic Cultures ◽  
Morphological Alterations ◽  
New Horizons ◽  
Immature Seeds ◽  
Cryopreservation Protocol

This study aimed to establish a cryopreservation protocol for embryogenic cultures of A. angustifolia, enabling the ex situ conservation of the species. Embryogenic cultures were established from immature seeds and treated with variations of the cryoprotectant solutions SuDG, SoD and PVS2 prior to immersion in liquid nitrogen. Cell viability was evaluated after 30, 60 and 90 days of re-growth. The highest re-growth without morphological alterations and with normal biochemical composition was obtained with the PVS2 solution with 40 min immersion in ethanol (-20 °C). This procedure opens new horizons for the ex situ conservation of the species genetic.

25 A preliminary study of the effects of breed and nanowater as extender diluent on ram semen characteristics post-thawing

2020 ◽  
Vol 32 (2) ◽  
pp. 138
Author(s):  
M. Murawski ◽  
J. Szymanowicz ◽  
T. Schwarz ◽  
P. Bartlewski
Keyword(s):  
Sperm Morphology ◽  
Egg Yolk ◽  
Plasma Generator ◽  
Deionized Water ◽  
Ex Situ ◽  
Survival Times ◽  
Becton Dickinson ◽  
Significant Difference ◽  
Semen Extender ◽  
Difference Test

This study examined the effect of genotype and extender diluent on the characteristics of frozen-thawed ram semen. Twenty ejaculates collected from 10 rams aged 4-12 years (2 Polish Lowland (PON), 4 synthetic line BCP (Berrichon du Cher×Charolais×PON/Polish Merino), and 4 SCP (Suffolk×Charolais×PON/Polish Merino) in mid-breeding season were divided into six equal portions each and frozen in semen extender prepared by mixing commercial Triladyl extender (MiniTub GmbH) with deionized water (Aqua Purificata, Prolab) or nanowater (NW; deionized water declusterized in a cold plasma generator for 15, 30, 45, 60, or 90 minutes (15’, 30’, 45’, 60’, or 90’, respectively); Nanotechnology Systems) and Gallus domesticus egg yolk (1:3:1 vol/vol/vol). All semen samples were evaluated for progressive motility (Sperm Class Analyzer) and sperm morphology (Nikon Eclipse 80i microscope, Nikon Corp.). In addition, the ex situ survival time at 37°C and extender content of ALT (alanine transferase), ALP (alkaline phosphatase), and aspartate aminotransferase were measured, and the numbers of apoptotic, necrotic, and live spermatozoa were determined post-thawing with flow cytometry (BD Accuri™ C6 Plus, Becton Dickinson). Data were analysed by two-way analysis of variance and least significant difference test (SigmaPlot, Systat Software Inc.). The PON exceeded (P<0.05) SCP rams in the occurrence of mid-piece and double tail defects using NW30’ and NW45’ extenders. The proportion of spermatozoa with proximal droplets was greater (P<0.05) in PON than in SCP or BCP rams with NW15’ and NW30’ extenders, respectively, but the percentage of sperm with detached heads was greater (P<0.05) in BCP and SCP compared with PON using NW15’ extender. Overall, spermatozoa in PON rams’ semen had survival times greater by 25-50min (P<0.05) compared with those for semen from BCP and SCP synthetic lines. The ALT levels were significantly elevated in SCP compared with PON and BCP rams only in NW90’ extender. The numbers of necrotic spermatozoa were greater (P<0.05) in SCP and BCP compared with PON rams, and the numbers of apoptotic spermatozoa were greater (P<0.05) in BCP than in SCP rams with NW45’, NW60’, and NW90’ extenders. Live spermatozoa were more prevalent (P<0.05) in PON than in BCP rams in NW45’ extender post-thawing. To summarise, semen from PON rams had higher survival times and fewer spermatozoa with detached heads but more mid-piece defects and sperm with double tails post-thawing compared with BCP and SCP breeds. The number of necrotic cells was greater in semen from BCP and SCP rams compared with PON. It can be concluded that breed-related differences in post-thaw sperm morphology were apparent in extenders containing NW15-60’, whereas the variations in cell necrosis/apoptosis were primarily seen with the use of NW45-90’ extenders.

scholarly journals Cryofreezing of Grass Carp (Ctenopharyngodon idella) Spermatozoa for Ex Situ Conservation

2013 ◽  
Vol 21 (1-2) ◽  
pp. 141-150 ◽  
Author(s):  
MRI Sarder ◽  
MA Hossain ◽  
MS Hossain
Keyword(s):  
Grass Carp ◽  
Egg Yolk ◽  
The Other ◽  
Ctenopharyngodon Idella ◽  
Ex Situ ◽  
Optimum Level ◽  
Dilution Ratio ◽  
Other Hand ◽  
Grass Carp Ctenopharyngodon Idella ◽  
Thaw Period

Experiments were conducted to develop and standardize the protocols for cryopreservation of sperm of grass carp (Ctenopharyngodon idella). Seven extenders such as Alsever’s solution, Urea egg-yolk, Egg-yolk citrate, Kurokura- 1, Kurokura-2, 0.9% NaCl and 0.6% Glucose and five cryoprotectants i.e. DMSO, methanol, ethanol, DMA and glycerol were employed for finding suitable combinations. Cryodiluents were prepared by mixing the cryoprotectants at 10% concentration of the extenders (v/v). Milt and cryodiluents were mixed at a ratio of 1:9 for Alsever’s solution, Kurokura-1, Kurokura-2, 0.9% NaCl and 0.6% glucose, and 1:4 for urea egg-yolk and egg-yolk citrate. Among the 35 combinations of extenders and cryoprotectants, Alsever’s solution with ethanol and methanol, urea egg-yolk and egg-yolk citrate with DMSO found suitable for preservation and it produced 74 ± 2.44%, 72 ± 2.54%, 76 ± 2.44% and 75 ± 2.23% spermatozoan motility at the post-thaw period respectively. Rest of the combinations, on the other hand, produced <60% motility at the post-thaw period and glycerol was found to be clotted after freezing. The dilution of milt with cryodiluent has been tested using six dilution ratios (1:2, 1:4, 1:7, 1:9, 1:15, 1:20) and found that 1:9 dilution ratio produced the highest post-thaw spermatozoan motility with Alsever’s solution (>75%) and 1:4 with urea eggyolk and egg-yolk citrate (>70%). As an optimum level of cryoprotectant, 10% concentration was found effective to produce significantly highest (P<0.05) percentage of spermatozoan motility compared to those of other four concentrations (5, 15, 20 and 30%).DOI: http://dx.doi.org/10.3329/pa.v21i1-2.16763 Progress. Agric. 21(1 & 2): 141 - 150, 2010

scholarly journals Effect of physiological levels of caffeine on Ca2+ handling and fatigue development in Xenopus isolated single myofibers

2009 ◽  
Vol 296 (5) ◽  
pp. R1512-R1517 ◽  
Author(s):  
Joelle I. Rosser ◽  
Brandon Walsh ◽  
Michael C. Hogan
Keyword(s):  
Generation Time ◽  
Relaxation Times ◽  
Ringer Solution ◽  
The Other ◽  
Force Generation ◽  
Muscle Performance ◽  
Tension Development ◽  
Fatigue Development ◽  
Significant Difference ◽  
Ergogenic Effects

The purpose of the present study was to determine whether exposure to exogenous physiological concentrations of caffeine influence contractility, Ca2+ handling, and fatigue development in isolated single Xenopus laevis skeletal muscle fibers. After isolation, two identical contractile periods (separated by 60-min rest) were conducted in each single myofiber ( n = 8) at 20°C. During the first contractile period, four fibers were perfused with a noncaffeinated Ringer solution, while the other four fibers were perfused with a caffeinated (70 μM) Ringer solution. The order was reversed for the second contractile period. The single myofibers were stimulated during each contractile period at increasing frequencies (0.16, 0.20, 0.25, 0.33, 0.50, and 1.0 tetanic contractions/s), with each stimulation frequency lasting 2 min until fatigue ensued, defined in this study as a fall in tension development to 66% of maximum. Tension development and free cytosolic [Ca2+] (fura-2 fluorescence spectroscopy) were simultaneously measured. There was no significant difference in the peak force generation, time to fatigue, cytosolic Ca2+ levels, or relaxation times between the noncaffeinated and caffeinated trials. These results demonstrate that physiological levels of caffeine have no significant effect on Xenopus single myofiber contractility, Ca2+ handling, and fatigue development, and suggest that any ergogenic effects of physiological levels of caffeine on muscle performance during contractions of moderate to high intensity are likely related to factors extraneous to the muscle fiber.

scholarly journals Comparison of growth performance between cryopreserved and fresh sperm-originated fry of Barbonymus gonionotus

1970 ◽  
Vol 7 (1) ◽  
pp. 145-149 ◽  
Author(s):  
F Rahman ◽  
MRI Sarder ◽  
MA Rouf
Keyword(s):  
Growth Performance ◽  
Egg Yolk ◽  
Citrate Solution ◽  
Control Groups ◽  
Significant Difference ◽  
Cryopreserved Sperm ◽  
Survival And Growth ◽  
And Control ◽  
Silver Barb ◽  
Glass Aquarium

This experiment was conducted to compare the growth performance of silver barb fry produced from cryopreserved sperm with those produced from fresh sperm. Cryopreserved sperm used for fry production was preserved with three extenders, Alsever's solution, urea egg-yolk, egg-yolk citrate solution and one cryoprotectant, DMSO. Cryodiluents were prepared by mixing the cryoprotectants at 10% concentration of the extender (% v/v). Fry produced with fresh sperm was considered as control. For comparing the growth, 60 fry of 15 day-old for each treatment of both cryopreserved and control groups were stocked to glass aquarium (50 cm x 30 cm x 28 cm) and reared them for ten weeks. Growth of fry in terms of length and weight increment in both cryopreserved and control groups were measured weekly. The growth pattern was more or less similar for all the treatments and there was no significant difference (P>0.05) between them. The survival rate of fry produced from cryopreserved sperm was 82.3% and that from fresh sperm 88%, and there was no significant difference (P>0.05) between two groups. It is therefore, concluded that the use of cryopreserved sperm does not impair survival and growth of fry. Keywords: Silver barb; Cryopreservation; Growth; Survival DOI: 10.3329/jbau.v7i1.4977 J. Bangladesh Agril. Univ. 7(1): 145-149, 2009

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