9 BIRTH OF CALVES AFTER ARTIFICIAL INSEMINATION WITH CRYOPRESERVED BOVINE EPIDIDYMAL SPERMATOZOA HARVESTED FROM POSTMORTEM BULLS

2009 ◽  
Vol 21 (1) ◽  
pp. 105 ◽  
Author(s):  
C. A. Guerrero ◽  
G. Gentry ◽  
J. Saenz ◽  
K. R. Bondioli ◽  
R. A. Godke
Keyword(s):  
Sperm Concentration ◽  
Egg Yolk ◽  
Beef Cows ◽  
Gestation Length ◽  
Epididymal Sperm ◽  
Average Birth Weight ◽  
Male Gametes ◽  
Potential Source ◽  
Mixed Breed ◽  
The Mean

Recently, interest in the preservation of epididymal sperm as a potential source of valuable genes for genome resource banks has increased. The ability to recover and cryopreserve postmortem epididymal sperm will allow the use of valuable gametes from a breeding male that dies unexpectedly. The objective of this study was to produce pregnancies and live births from AI by using frozen–thawed bovine caudal epididymal sperm. Paired testes were obtained from mature, mixed-breed beef bulls (n = 3) from a local abattoir and transported to the laboratory (within 5 h postmortem) in a Styrofoam box at a temperature that ranged between 28 and 29°C. The mean ± SEM weights of the pair of testes and caudae epididymides were 345 ± 9g and 10 ± 2g, respectively. Epididymal sperm from each bull were harvested by multiple incisions from the caudae epididymides, pooled, and then rinsed with 4 mL of egg yolk Tris-glucose citric acid monohydrate extender (EYT-GC). Harvested sperm samples were placed in a refrigerator at 4°C for 4 h before the addition of cryoprotectant. Samples were diluted slowly over a period of 30 min 1:1 in EYT-GC extender containing 14% glycerol to obtain a final concentration of 7% glycerol. Sperm concentration was adjusted to 35 million/straw and loaded into previously cooled 0.5-mL plastic straws. Straws were placed 2 cm above liquid nitrogen (LN2) vapors for 10 min and then plunged into LN2. Sperm were stored for 1 year before thawing. Luteal-phase crossbred beef cows were synchronized with a single 25-mg injection (i.m.) of prostaglandin, and those displaying standing estrus (n = 6) were each inseminated with 1 straw by a single technician from 1 bull 12 to 18 h after the onset of standing estrus. Straws were thawed for AI in a 37°C water bath for 40 s. Three of the 6 cows inseminated (50%) were diagnosed as pregnant by transrectal ultrasonography at 45 days post-AI. All 3 pregnant cows (100%) delivered healthy singleton calves (2 males and 1 female, with an average birth weight of 37 ± 2.5 kg), resulting in a mean gestation length of 286 ± 1.9 days (range: 282–288 days). We can conclude that epididymal sperm can be extracted by 5 h postmortem from bull testes, frozen, and subsequently used for the production of live offspring from AI. Further research is needed to improve this technology to optimize the utilization of valuable bovine male gametes.

215 EPIDIDYMAL SPERM CRYOPRESERVATION OF ONE SOMALIA WILD ASS (EQUUS AFRICANUS SOMALIENSIS) USING SIX DIFFERENT EXTENDERS

2006 ◽  
Vol 18 (2) ◽  
pp. 215 ◽  
Author(s):  
M. Álvarez ◽  
F. Martínez-Pastor ◽  
V. García-Macías ◽  
S. Borragán ◽  
M. Celada ◽  
...  
Keyword(s):  
Species Differences ◽  
Sperm Concentration ◽  
Egg Yolk ◽  
Red List ◽  
Biological Resource ◽  
Epididymal Sperm ◽  
Sperm Cryopreservation ◽  
The Poor ◽  
Wild Ass ◽  
Viable Sperm

The Somalia wild ass (Equus africanus somaliensis) is a critically endangered taxon (IUCN 2004 red list) which could benefit from biological resource banking. In this work, we studied the effect of different extenders applied to the cryopreservation of epididymal sperm obtained from one male of this subspecies. This animal (13 years old; housed in Cabarceno Park, Cantabria, Spain) was castrated because of very aggressive behavior with other mature males. Genitalia were dissected and weighed (testicles: right, 166 g, and left, 179 g; cauda epididymis: right, 9.3 g, and left, 11.8 g). Sperm were flushed from the cauda epididymis, yielding 15 mL of sample. Sperm concentration was 15 × 109 spermatozoa/mL, totaling 225 × 109 (allowing 4500 doses at 50 × 106 sperm/dose). Sperm motility (TM = % total motile; PM = % progressive; VAP = average path velocity) was assessed by CASA (Microptic, Barcelona, Spain). Viability (VIAB = % viable sperm) and acrosomal status (ACR = % viable spermatozoa with intact acrosomes) were assessed using propidium iodide (37 μmol/L) and PNA-FITC (1 ng/L) and flow cytometry. Chemicals were purchased from Sigma (Madrid, Spain). Part of the sample was divided into six aliquots and diluted 1:1 with different extenders: UL4: Tes-Tris-Fructose (TTF), 10% egg yolk (EG), and 4% glycerol (G); UL8: TTF, 20% EG, and 8% G; AND4: Andromed® (Minitüb, Tiefenbach, Germany) and 4% G; AND7: Andromed® and 7% G; GENT: Gent 1045; and INRA: INRA96 and 4% G. Andromed, Gent, and INRA are commercial extenders. Samples were cooled to 5°C (−0.2°C/min) and then diluted to 200 × 106 sperm/mL. Samples were packed (0.5-mL straws) and frozen using a biofreezer (from 5°C to −15°C at −15°C/min, and from −15°C to −100°C at −25°C/min). Samples were thawed at 65°C for 6 s, and assessed as for pre-freezing (Table 1). Post-thawing motility recovery using AND7 was excellent. The highest viability recovery was achieved by UL4, although that in AND7 was similar. The poor results of equine commercial extender Gent 1045 in this species are remarkable. Our results highlight the importance of species differences in the field of sperm cryopreservation. It is necessary to carry out continuous research for optimizing cryopreservation protocols in order to create germplasm banks for wild species. Table 1. Quality assessment results

160 FREEZABILITY OF ELECTROEJACULATED AND EPIDIDYMAL SPERMATOZOA FROM BLUE WILDEBEEST (CONNOCHAETES TAURINUS)

2009 ◽  
Vol 21 (1) ◽  
pp. 179
Author(s):  
M. Mata-Campuzano ◽  
M. Alvarez ◽  
S. Borragan ◽  
F. Martinez-Pastor ◽  
M. Nicolas ◽  
...  
Keyword(s):  
Behavioral Problems ◽  
Sperm Concentration ◽  
Egg Yolk ◽  
Domestic Species ◽  
Ruminant Species ◽  
Male Gametes ◽  
Nature Park ◽  
Sample Recovery ◽  
Epididymal Spermatozoa ◽  
Connochaetes Taurinus

Maintenance of population viability, especially endangered species, requires preservation of as much genetic variability as possible, therefore genetic resource banks are very important. For male gametes, preservation of all available sources (ejaculates and epididymal) are useful. Information regarding sperm characteristics of most wild ruminant species is limited compared to that from domestic species. The objective of this work was to characterize the freezability of electroejaculated and epididymal spermatozoa from a wildebeest (5 year old; housed in Cabárceno Nature Park, Cantabria, Spain) that was castrated because of behavioral problems. After general anesthesia (ethorfine + xilazine, 1.8 mL + 0.5 mg kg–1) with dart, semen was collected by electroejaculation (3 V and 75 mA). Sperm concentration was 250 × 106 mL–1 (total spermatozoa: 1128.6 × 106). After castration, epididymides were disected and spermatozoa were collected by making several incisions in the caudal epididymis. Concentration was 12 441 × 106 spermatozoa mL–1 (total spermatozoa: 24 882 × 106). Samples were diluted to 200 × 106 spermatozoa mL–1 (TesT-Fructose-Egg yolk-Glycerol-Antibiotics) and chilled to 5°C during 2 h. Diluted semen was packaged in 0.25-mL straws and frozen from 5°C to –100°C (–20°C min–1) in a programmable cell freezer (Kryo 10, Planer). Straws were plunged into liquid nitrogen until analysis and thawed in a water bath (65°C, 6 s). Fresh, pre-freezing and post-thawed samples were analysed for motility (total motility TM, %; progressive motility PM, %; path velocity VAP, μm s–1; track speed VCL, μm s–1; progressive velocity VSL, μm s–1) using a CASA (ISAS, Proiser, Valencia, Spain). Viability (VIAB %) (SYBR-14 and propidium iodide) and mitochondrial membrane potential (MIT %) (JC1) were assessed by flow cytometry. Post-thawing results for electroejaculated v. epididymal samples were, respectively: TM: 87.0 v. 64.6%; PM: 68.7 v. 33.4%; VAP: 95.9 v. 49.8 μm s–1; VCL: 108.3 v. 71.6 μm s–1; VSL: 86.7 v. 40.2 μm s–1; VIAB: 57.0 v. 73.9%; MIT: 59.5 v. 77.5%. Motility parameters were higher for the electroejaculated sample; however, viability was higher for the epididymal sample. Recovery rates (post-thawed value/pre-freezing value × 100) for electroejaculated v. epididymal samples were: TM: 97.2 v. 93.4%; PM: 113.3 v. 103.2%; VAP: 88.9 v. 122.0 μm s–1; VCL: 87.5 v. 126.0 μm s–1; VSL: 93.4 v. 125.3 μm s–1; VIAB: 75.0 v. 97.7%; MIT: 69.2 v. 95.9%). These rates suggest a good freezability of electroejaculated and epididymal spermatozoa in blue wildebeest. This work was supported in part by Cantur. 3 Supported by Juan de la Cierva program (MICINN, Spain).

scholarly journals Viability of carabao (Bubalus bubalis carabanensis) epididymal sperm from post mortem testes in semen extender at refrigerated temperature

2017 ◽  
Vol 4 (2) ◽  
pp. 5
Author(s):  
Lean Angelo B. San Diego ◽  
Marvin Bryan S. Salinas ◽  
Angelica C. Bumanlag ◽  
Marlon B. Ocampo ◽  
Lerma C. Ocampo
Keyword(s):  
Ambient Temperature ◽  
Sperm Concentration ◽  
Bubalus Bubalis ◽  
Post Mortem ◽  
Epididymal Sperm ◽  
Progressive Motility ◽  
Animal Genetic Resources ◽  
The Mean ◽  
Semen Extender ◽  
Excellent Source

Post mortem epididymal sperm (ES) is an excellent source of germplasm for conservation of animal genetic resources. In this study,  the viability of post mortem ES from carabao after collection at ambient temperature (AT) and after 24 hr and 48 hr of storage at refrigeration temperature (RT) were evaluated. ES were collected through slicing of epididymides from 7 carabaos. The mean ES volume was 0.4 ml with sperm concentration of 2.5×109 cells/ml. The mean percentage livability of fresh ES, after 24 hr and 48 hr of storage at RT were 81.93%, 65.93% and 43.7%, respectively. The mean percentage abnormalities of fresh ES, after 24 hr and 48 hr of storage at RT were 44.15%, 38.5% and 43.47%, respectively. The mean percentage motility of fresh ES, after 24 hr and 48 hr of storage at RT were 60.0%, 50.0% and 17.24%, respectively, after analysis through conventional means. Through CASA, the mean percentage motility was observed to be lower significantly at 26.12%, 37.94% and 18.32% for fresh ES, after 24 hr and 48 hr of storage at RT, respectively. The mean progressive motility of fresh ES, after 24 hr and 48 hr storage at RT were 10.02%, 16.47% and 10.87%, respectively. The results suggest that ES potential for use in fertilization studies remained viable when used immediately or after storage at RT for the first 24 hr.

Characteristics and freezability of Assam Hill goat semen

2017 ◽  
Author(s):  
S. Deori ◽  
B. C. Deka ◽  
R. K. Biswas ◽  
N. Nahardeka ◽  
A. Arangasamy ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Sperm Concentration ◽  
Egg Yolk ◽  
Mean Values ◽  
Acceptable Quality ◽  
Sperm Abnormality ◽  
Semen Characteristics ◽  
The Mean ◽  
Live Sperm

Assam Hill goat (AHG) is an important goat germplasm found in Assam and its adjoining areas of India. The study was designed with an objective to study the semen characteristics and freezability of AHG buck semen using Tris -Egg yolk-Citrate-Fructose diluent. The mean values of fresh semen characteristics in AHG bucks viz., ejaculate volume (ml), initial sperm motility (%), sperm concentration (x106/ml), live sperm (%), sperm abnormality (%), HOST-reacted sperm (%) and intact acrosome (%) recorded were 0.39 ± 0.01, 77.97 ± 0.73, 3201.00 ± 143.78, 83.02 ± 0.65, 7.66 ± 0.73, 66.95 ± 0.74 and 93.34 ± 0.51, respectively. Mean values for post-thaw semen characteristics i.e., sperm motility (%), live sperm (%), HOST-reacted sperm (%) and intact acrosome (%) were 55.39 ± 0.97, 71.01 ± 0.78, 54.77 ± 0.55 and 82.16 ± 0.43, respectively. It can be concluded that AHG bucks donate acceptable quality of semen which can be frozen successfully in Tris-Egg yolk-Citrate-Fructose diluents for using in Artificial Insemination.

scholarly journals EPIDIDYMAL SPERMATOZOA QUALITY OF ETAWA CROSSBREED GOAT IN TRIS EXTENDER SUPPLEMENTED WITH VARIOUS LACTOSE CONCENTRATIONS

2017 ◽  
Vol 11 (1) ◽  
pp. 15-18
Author(s):  
Muhammad Riyadhi ◽  
Agus Setiawan ◽  
Herdis Herdis ◽  
Muhammad Rizal
Keyword(s):  
Sperm Concentration ◽  
Egg Yolk ◽  
Control Group ◽  
Spermatozoa Motility ◽  
The Mean ◽  
Epididymal Spermatozoa ◽  
Live Sperm

This research was conducted to investigate the effect of various concentrations of lactose supplementation in Tris extender for maintaining the quality of Etawa crossbreed goat epididymal spermatozoa stored at 3-5° C. Semen in the control group was diluted with a tris extender containing 20% egg yolk without lactose. Semen in the test groups was diluted with a tris extender containing 20% egg yolk and added with 0.3% (0.3 g per 100 mL extender) and 0.6% lactose for group TL1 and TL2, respectively. Parameters evaluated of the fresh epididymal spermatozoa were motility, concentration, percentage of live, and abnormality of spermatozoa, while for diluted-spermatozoa were motility and percentage of live spermatozoa. Spermatozoa observation was conducted until it reaches 40% motility. The results showed that the mean percentage of motility, live sperm, concentration, and abnormality of epididymal spermatozoa were 70%; 81%; 3,220x106 cells/mL; and 4.30%, respectively in all group. After dilution, the percentage of motility and live spermatozoa were also 70% and 81.00±1.58%, respectively in all groups. The decreasing of spermatozoa motility was observed on day 4 of storage, in which percentage of spermatozoa motility in control group (40.00±0.00%) was significantly lower (P<0.05) than those in TL1 (44.00±2.24%) and TL2(45.00±0.00%) groups. Percentage of live spermatozoa in control (63.20±2.68%) was not significantly different (P>0.05) than TL1(65.40±1.95%) and TL2 (65.60±1.95%). In conclusion, the supplementation of lactose into Tris extender could maintain the epididymal spermatozoa of Etawa crossbreed for 3 days of storage at 3-5° C.

Effect of Tomato Juice in Tris-Yolk-Citrate Extender onRefrigeration Preservation of Cauda Epididymal Spermatozoa of Buck

2019 ◽  
Vol 15 (01) ◽  
pp. 71-74
Author(s):  
L M Chaudhary ◽  
C T Khasatiya ◽  
Amarjeet Amarjeet ◽  
A B Yede
Keyword(s):  
Adverse Effect ◽  
Egg Yolk ◽  
Physical Characteristics ◽  
Tomato Juice ◽  
The Mean ◽  
Epididymal Spermatozoa ◽  
Sperm Traits

The study was carried out on the preservation of epididymal spermatozoa of buck at refrigerated temperature without and with tomato juice as a supplement in Tris egg yolk citrate extender. The eight pairs of testicles including epididymis (total 16) from slaughtered bucks were collected within 2–4 hours of their slaughter. Sperms collected from cauda epididymis were preserved at refrigerated temperature up to 48 hours in tris egg yolk citrate extender at 300 million sperm/mL with different concentration of tomato juice (0%, 4%, 6%, 8%, and 10%) and the physical characteristics of spermatozoa were assessed to know the effect of tomato juice (Tj). The mean dead, abnormal and HOST non-reacted spermatozoa increased significantly (p less than0.05) at every 12-hour intervals of preservation in the dilutor without and with different concentration of tomato juice. Tomato juice exerted an adverse effect on physical characteristics of sperm during refrigeration preservation. All the three sperm traits studied however revealed significant (p less thaN 0.01) positive interrelationships with correlations of 0.31 to 0.72.

The effects of Pb on sperm parameters and sperm DNA fragmentation of men investigated for infertility

2020 ◽  
Vol 31 (4) ◽  
Author(s):  
G.U.S. Wijesekara ◽  
D.M.S. Fernando ◽  
S. Wijeratne
Keyword(s):  
Dna Fragmentation ◽  
Seminal Plasma ◽  
Sperm Concentration ◽  
Sperm Parameters ◽  
World Health ◽  
Sperm Dna Fragmentation ◽  
Progressive Motility ◽  
Sperm Dna ◽  
The Mean ◽  
Pb Concentration

AbstractBackgroundLead (Pb) is one of the metals most prevalent in the environment and is known to cause infertility and deoxyribonucleic acid (DNA) fragmentation. This study aimed to determine the association between seminal plasma Pb and sperm DNA fragmentation in men investigated for infertility.MethodsMale partners (n = 300) of couples investigated for infertility were recruited after informed consent was obtained. Sperm parameters were assessed according to the World Health Organization (WHO) guidelines. Seminal plasma Pb was estimated by atomic absorption spectrophotometry after digestion with nitric acid.ResultsIn Pb-positive and -negative groups the sperm parameters and sperm DNA fragmentation were compared using independent sample t-test and the Mann-Whitney U-test, respectively. The mean [standard deviation (SD)] age and duration of infertility were 34.8 (5.34) years and 45.7 (35.09) months, respectively, and the mean Pb concentration was 15.7 μg/dL. In Pb positives compared to Pb negatives the means (SD) of sperm count, progressive motility viability and normal morphology were lower (p > 0.05) but the DNA fragmentation was significantly higher 39.80% (25.08) than Pb negatives 22.65% (11.30). Seminal plasma Pb concentration and sperm DNA fragmentation had a positive correlation (r = 0.38, p = 0.03). A negative correlation was observed between sperm DNA fragmentation and sperm concentration, progressive motility, total motility and viability. When the DNA fragmentation was ≥30% sperm concentration and viability decreased (p < 0.05).ConclusionsPb in seminal plasma had a significant effect on sperm DNA fragmentation but not with other sperm parameters.

Cryopreservation of Spix's yellow-toothed cavy epididymal sperm using Tris- and coconut water-based extenders supplemented with egg yolk or Aloe vera

Cryobiology ◽  
2021 ◽  
Author(s):  
Samara Sandy Jeronimo Moreira ◽  
Andreia Maria da Silva ◽  
Ana Liza Paz Souza ◽  
Erica Camila Gurgel Praxedes ◽  
João Batista Freire de Souza Junior ◽  
...  
Keyword(s):  
Aloe Vera ◽  
Egg Yolk ◽  
Coconut Water ◽  
Epididymal Sperm ◽  
Water Based

scholarly journals 158 Provision of artificial shade before weaning influences performance of Angus and Brangus cow-calf pairs

2020 ◽  
Vol 98 (Supplement_4) ◽  
pp. 129-130
Author(s):  
Gleise Medeiros da Silva ◽  
Tessa M Schulmeister ◽  
Federico Podversich ◽  
Federico Tarnonsky ◽  
Maria E Zamora ◽  
...  
Keyword(s):  
Body Condition Score ◽  
Average Daily Gain ◽  
Beef Cows ◽  
Paspalum Notatum ◽  
Gestation Length ◽  
Long Term Effects ◽  
Weaning Weight ◽  
Randomized Design ◽  
Condition Score ◽  
The Impact

Abstract A completely randomized design study with a 2 × 2 factorial arrangement of treatments evaluated the impact of artificial shade (SHADE or NO SHADE) and breed (ANGUS vs. BRANGUS) on performance of cows, nursing calves, and subsequent offspring. Twenty-four Angus and 24 Brangus black-hided pregnant cows (579 ± 8 kg BW; 6.5 yr; approximately 85 d of gestation) and their nursing calves were randomly allocated to 12 ‘Pensacola’ bahiagrass pastures (Paspalum notatum Flüggé; 1.3 ha; n = 4 pairs/pasture), with or without access to artificial shade (NO SHADE BRANGUS [NSB], NO SHADE ANGUS [NSA], SHADE BRANGUS [SB], and SHADE ANGUS [SA]) for 56 d during summer. Body condition score (BCS) of cows and BW of pairs were obtained on d -1, 0, 55, and 56 (weaning weight). Following weaning, calves were randomly allocated to 4 pens (n = 12/pen) equipped with GrowSafe feed bunks for 14 d to measure feed intake (DMI) and efficiency (G:F). A shade × breed interaction (P &lt; 0.05) was observed for average daily gain (ADG) and BCS of cows, with SB being greatest (P ≤ 0.05). Pre-weaning calf ADG tended to be greater (P = 0.10) for SHADE vs. NO SHADE. Weaning weight and BW 14-d post-weaning were lesser for NSB vs. NSA, SA, and SB, whereas no differences in DMI, ADG, or G:F were observed (P &gt; 0.11). Gestation length was greater for SHADE vs. NO SHADE cows (292 vs. 274; P = 0.02), but calf birth weight was not different. Providing artificial shade to pregnant-lactating beef cows positively impacted the growth of Brangus but not Angus cows. However, weaning BW of calves from Angus cows regardless of shade access did not differ from that of Brangus calves with shade. Further research should investigate the potential long-term effects of shade on the subsequent offspring.

Surface culture of Steinernema sp. on two solid media and their pathogenicity against Galleria mellonella

2021 ◽  
Vol 95 ◽  
Author(s):  
C.I. Cortés-Martínez ◽  
A.I. Rodríguez-Hernández ◽  
M.R. López-Cuellar ◽  
N. Chavarría-Hernández
Keyword(s):  
Galleria Mellonella ◽  
Egg Yolk ◽  
Infective Juvenile ◽  
Surface Culture ◽  
Solid Media ◽  
Significant Difference ◽  
Bacterial Lawn ◽  
The Mean

Abstract The use of native entomopathogenic nematodes as biocontrol agents is a strategy to decrease the environmental impact of insecticides and achieve sustainable agriculture crops. In this study, the effect of the surface culture of Steinernema sp. JAP1 over two solid media at 23–27°C on infective juvenile (IJ) production and pathogenicity against Galleria mellonella larvae were investigated. First, the bacterial lawn on the surface of the media with egg yolk (P2) or chicken liver (Cl) were incubated in darkness at 30°C for 48 and 72 h, and 100 surface-sterilized IJs were added. Four harvests were conducted within the next 35 days and the mean accumulated production was superior on Cl (210 × 103 IJs) than on P2 (135 × 103 IJs), but the productivity decreased up to 10% when the incubation time of the bacterial lawn was of 72 h. The mean pathogenicity of in vitro- and in vivo-produced IJs were of 47–64% and 31%, respectively. It is worth noting that none of the two solid media had a statistically significant difference in IJ pathogenicity. Considering that the maximum multiplication factor of IJs on solid media was 2108 and that the pathogenicity against G. mellonella was outstanding, Steinernema sp. has a good potential for in vitro mass production.

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