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).

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.

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.

Assessment of Pomegranate Juice as Antioxidant in Extender on Cauda Epididymis Spermatozoal Quality of Buck at Refrigerated Temperature

2019 ◽  
Vol 15 (02) ◽  
pp. 26-29
Author(s):  
Amarjeet Amarjeet ◽  
C T Khasatiya ◽  
L Chaudhary
Keyword(s):  
Detrimental Effect ◽  
Egg Yolk ◽  
Antioxidant Effect ◽  
Pomegranate Juice ◽  
Antioxidant Additive ◽  
Epididymal Spermatozoa ◽  
The Dead

The present investigation was carried out to study the refrigeration preservation of the cauda epididymal retrieved spermatozoa of buck in Tris egg yolk citrate (TEYC) dilutor containing pomegranate juice as antioxidant additive. The retrieved cauda epididymal spermatozoa extended in TEYC dilutor were studied in five groups by adding different concentration of pomegranate juice as additive (0% as control T1 group and 5%, 10%, 15% and 20% as treatment T2, T3, T4 and T5 groups, respectively) and storing at refrigerated temperature up to 48 hr. The results showed that the control extender had the least dead, abnormal and HOS non-reacted sperm percent among all treatments tested and that with increasing the pomegranate juice concentration in dilutor, the percentage of the dead, abnormal and HOST non-reacted spermatozoa increased significantly. The same trend was observed at all 12 hourly storage intervals indicating its detrimental effect on epididymal sperms of bucks at refrigeration temperature. The dead, abnormal, and HOST non-reacted sperm were significantly and positively interrelated with each other (r = 0.53-0.83). It was concluded that the inclusion of pomegranate juice in TEYC dilutor did not show any beneficial/antioxidant effect on epididymal sperms of buck in fresh or refrigerated semen and in fact all the levels of pomegranate juice (5% to 20%) were detrimental to cauda epididymal spermatozoa of a buck.

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.

scholarly journals 209FUNCTIONAL ASSESSMENT OF WHITE RHINOCEROS CERATHOTERIUM SIMUM EPIDIDYMAL SPERMATOZOA BEFORE AND AFTER CRYOPRESERVATION

2004 ◽  
Vol 16 (2) ◽  
pp. 226 ◽  
Author(s):  
F. Martinez-Pastor ◽  
F. Olivier ◽  
T. Spies ◽  
L. Anel ◽  
P. Bartels
Keyword(s):  
Plasma Membrane ◽  
Assisted Reproduction ◽  
Phase Contrast ◽  
Egg Yolk ◽  
Phase Contrast Microscopy ◽  
White Rhinoceros ◽  
Contrast Microscopy ◽  
Before And After ◽  
Epididymal Spermatozoa

Biological Resource Banks represent a potentially valuable tool for species conservation. It is, however, necessary to understand the species-specific cryopreservation process and its consequences for spermatozoa to aid in the development of assisted reproduction as a future conservation tool. The aim of this study was to assess the in vitro functionality of white rhinoceros Cerathoterium simum epididymal spermatozoa both before and after cryopreservation. Testes from a harvested white rhino bull were removed and transported at 5°C to the laboratory within 4h. The cauda epididymis was dissected out and flushed with 2mL of Tris-citrate egg yolk extender (fraction A, Biladyl, Minitüb, Germany). A 0.1mL aliquot was removed for analysis and the balance (9mL; 2mL fraction A+7mL sperm sample) mixed with an additional 27.2mL of Tris-citrate egg yolk with glycerol (fraction B, Bidadyl). The extended sample was allowed to cool to 4°C over a 6-h period before an additional 29.2mL of cooled fraction B were added (final sperm concentration=150×106mL−1). Sperm samples were loaded into 0.25-mL straws and frozen over LN2 vapor (4cm for 20min) for later assessment. Sperm straws were thawed by placing the straws in water at 37°C for 30s. Pre-freeze and post-thaw evaluations were carried out in the same manner. Media used included: HEPES for washing (20mM HEPES, 355mM sucrose, 10mM glucose, 2.5mM KOH) and HEPES saline (197mM NaCl, instead of sucrose). An aliquot was diluted with HEPES (washing) and centrifuged for 5min at 600×g; the pellet was resuspended in HEPES saline. Sperm motility (total motility %, TM;; and progressive motility %, PM) was assessed using phase contrast microscopy (×200; 37°C). Sperm plasma membrane status was assessed using the fluorescent dye, propidium iodide (50ngmL−1 in HEPES saline;; 10min, RT). Percentage of cells with plasma membranes intact (unstained;; PMI) was recorded. Mitochondrial status was assessed with the fluorescent dye, JC-1 (7.5μM in HEPES saline;; 30min, 37°C). The % of cells with an orange-stained midpiece was recorded (active mitochondria;; MIT). Resilience to hypoosmotic shock (HOS test) was assessed by diluting a sample in 100mOsm/kg HEPES saline (1:20; 15min, RT). An aliquot was stained with PI to assess plasma membrane status (HOSPMI), and the rest was fixed with formaldehyde, and % coiled tails (positive endosmosis;; HOST) was estimated using phase contrast microscopy (×400). Evaluations of PMI, MIT and HOSPMI were performed using fluorescence microscopy (×400, 450–490nm excitation filter). The results indicated that quality was good pre-freezing (TM: 60%; PMI: 86%; MIT: 100%), except for a PM value of 15%. After thawing, although there was a drop in TM (30%), there was no decrease in PM (20%). Our in vitro functional assessment indicated a loss of quality between the pre-freeze and post-thaw evaluations, but PMI and MIT maintained their pre-thaw levels (60% and 72%, respectively). The HOS test, which indicates plasma membrane integrity, decreased from the pre-freeze level (91%) to a post-thaw value of 70%. HOSTPMI was 72% pre-freeze, and decreased to 54% post-thaw. In conclusion, epididymal spermatozoa from the white rhino may retain its functionality after cryopreservation in a commerically available cryo-extender (Bidadyl). The use of assisted reproduction techniques could someday play a role in the management and conservation of the white rhinoceros and related species.

Interpreting the diet of extinct ruminants: the case of a non-browsing giraffid

Paleobiology ◽  
1988 ◽  
Vol 14 (3) ◽  
pp. 287-300 ◽  
Author(s):  
Nikos Solounias ◽  
Mark Teaford ◽  
Alan Walker
Keyword(s):  
Shape Analysis ◽  
Feeding Habits ◽  
African Buffalo ◽  
Giraffa Camelopardalis ◽  
Ruminant Species ◽  
Modern Species ◽  
Quantitative Analyses ◽  
The Mean ◽  
Connochaetes Taurinus ◽  
Rule Out

Modern ruminant species have traditionally been placed in three broad dietary categories (browsers, grazers and intermediates) based on their observed feeding habits. Shape analysis of premaxillary outlines of 31 species of ruminants shows that their premaxillae differ according to their dietary category. Browsers have pointed premaxillae and grazers square ones. Intermediate feeders have intermediate outlines.The Miocene giraffid Samotherium boissieri has always been viewed as a specialized browser similar to the modern okapi, Okapia johnstoni. However, the premaxillary shape of S. boissieri falls very close to the mean of the grazers and is most similar to that of the African buffalo (Syncerus caffer), a committed grazer.Quantitative analyses of the microscopic wear patterns on the molars reveal significant differences between three modern species from the three dietary groups. S. boissieri has more microscopic scratches on its teeth than either the modern giraffe Giraffa camelopardalis (a browser) or Grant's gazelle Gazella granti (an intermediate feeder). In this respect, it is indistinguishable from the wildebeest Connochaetes taurinus which is a committed grazer.Both of these analyses suggest that this extinct giraffid was a grazer, although we cannot rule out the possibility that it was an intermediate feeder. It was definitely not a specialized browser as are both living members of the Giraffidae.

scholarly journals The expansion of the TRB and TRG genes in domestic goats (Capra hircus) is characteristic of the ruminant species

BMC Genomics ◽  
2020 ◽  
Vol 21 (1) ◽  
Author(s):  
Francesco Giannico ◽  
Serafina Massari ◽  
Anna Caputi Jambrenghi ◽  
Adriano Soriano ◽  
Angela Pala ◽  
...  
Keyword(s):  
T Cell ◽  
T Cell Receptor ◽  
Genomic Sequence ◽  
Genomic Structure ◽  
Genetic Research ◽  
Cell Receptor ◽  
Capra Hircus ◽  
Meat Production ◽  
Domestic Species ◽  
Ruminant Species

Abstract Background Goats (Capra hircus), one of the first domesticated species, are economically important for milk and meat production, and their broad geographical distribution reflects their successful adaptation to diverse environmental conditions. Despite the relevance of this species, the genetic research on the goat traits is limited compared to other domestic species. Thanks to the latest goat reference genomic sequence (ARS1), which is considered to be one of the most continuous assemblies in livestock, we deduced the genomic structure of the T cell receptor beta (TRB) and gamma (TRG) loci in this ruminant species. Results Our analyses revealed that although the organization of the goat TRB locus is broadly similar to that of the other artiodactyl species, with three in-tandem D-J-C clusters located at the 3′ end, a complex and extensive series of duplications have occurred in the V genes at the 5′ end, leading to a marked expansion in the number of the TRBV genes. This phenomenon appears to be a feature of the ruminant lineage since similar gene expansions have also occurred in sheep and cattle. Likewise, the general organization of the goat TRG genes is typical of ruminant species studied so far, with two paralogous TRG loci, TRG1 and TRG2, located in two distinct and distant positions on the same chromosome as result of a split in the ancestral locus. Each TRG locus consists of reiterated V-J-J-C cassettes, with the goat TRG2 containing an additional cassette relative to the corresponding sheep and cattle loci. Conclusions Taken together, these findings demonstrate that strong evolutionary pressures in the ruminant lineage have selected for the development of enlarged sets of TRB and TRG genes that contribute to a diverse T cell receptor repertoire. However, differences observed among the goat, sheep and cattle TRB and TRG genes indicate that distinct evolutionary histories, with independent expansions and/or contractions, have also affected each ruminant species.

scholarly journals Evaluation of Modified BACTEC 12B Radiometric Medium and Solid Media for Culture of Mycobacterium aviumsubsp. paratuberculosis from Sheep

1999 ◽  
Vol 37 (4) ◽  
pp. 1077-1083 ◽  
Author(s):  
R. J. Whittington ◽  
I. Marsh ◽  
S. McAllister ◽  
M. J. Turner ◽  
D. J. Marshall ◽  
...  
Keyword(s):  
Egg Yolk ◽  
Standard Test ◽  
Johne's Disease ◽  
Johne’S Disease ◽  
Media Culture ◽  
Ruminant Species ◽  
Solid Media ◽  
Wide Range ◽  
Causative Bacterium ◽  
Culture Positive

Definitive diagnosis of Johne’s disease in ruminants depends on confirming the presence of the causative bacterium, Mycobacterium avium subsp. paratuberculosis, in tissues of the host. This is readily achieved in most ruminant species by culture. However, culture of clinical specimens from sheep in many countries has been unrewarding. Such a culture from sheep was achieved recently in Australia by using a radiometric culture medium. The aims of the present study were to evaluate the culture of M. aviumsubsp. paratuberculosis from sheep by using modified BACTEC 12B radiometric medium, to determine the sensitivity of culture in relation to histopathology, and to evaluate a range of solid media. Culture of M. avium subsp. paratuberculosisfrom sheep with Johne’s disease is a sensitive method of diagnosis: intestinal tissues from all 43 animals with multibacillary disease and all 22 animals with paucibacillary disease were culture positive, while 98% of feces from 53 animals with multibacillary disease and 48% of feces from 31 animals with paucibacillary disease were culture positive. Of sheep without histological evidence of Johne’s disease from infected flocks, intestinal tissue from 32% of 41 were culture positive, while feces from 17% of 41 were culture positive. Consequently, culture is recommended as the “gold standard” test for detection of ovine Johne’s disease. Of the wide range of solid media that were evaluated, only modified Middlebrook 7H10 and 7H11 agars, which were very similar in composition to modified BACTEC 12B medium, yielded growth of ovine strains of M. avium subsp.paratuberculosis. The sensitivity of detection of M. avium subsp. paratuberculosis on solid media was slightly lower than that in modified BACTEC 12B radiometric medium. Both egg yolk and mycobactin J were essential additives for growth of ovine strains of M. avium subsp.paratuberculosis in both liquid and solid media.

scholarly journals Effect of split ejaculation and seminal extenders on longevity of donkey semen preserved at 5° C

2000 ◽  
Vol 52 (4) ◽  
pp. 372-378 ◽  
Author(s):  
S.L.V. Mello ◽  
M. Henry ◽  
M.C. Souza ◽  
S.M.P. Oliveira
Keyword(s):  
Phase Contrast ◽  
Room Temperature ◽  
Sperm Morphology ◽  
Sperm Concentration ◽  
Egg Yolk ◽  
The Rich ◽  
Student’S T ◽  
The One ◽  
Artificial Vagina ◽  
Student’S T Test

The aim of this study was to evaluate the longevity of donkey sperm comparing the rich seminal fraction and the whole semen in two extenders, Kenney and modified Baken extenders. Semen of five donkeys were collected through an open-end artificial vagina once a week for five consecutive weeks. The two first jets (rich fraction) of semen were collected separately from the rest of the ejaculate. Whole semen samples were obtained mixing proportionally part of the rich with part of the poor seminal fractions. Seminal samples were immediately diluted 1:1 in each extender and maintained at room temperature during sperm concentration analysis. Samples were further diluted to rich 50×10(6) sperm per ml, cooled in a refrigerator at the initial rate of -0.6° C/min and preserved at 5° C. Total motility (TM), progressive motility (PM) and sperm vigor (V) were examined after final dilution and cooling, and every 24 hours up to the decrease of total motility under 10%. Sperm morphology was evaluated using a phase contrast microscope directly after dilution, on days 3, 6 and 9 post collection. It was used a 2×2 factorial design in a randomised bloc experiment, and means were compared by Student’s t test. Longevity did not vary between the rich seminal fraction and the whole semen for both extenders used. TM, PM, V and sperm morphology were better preserved in the extender with egg yolk (modified Baken extender) than in the one with skimmed milk (Kenney) in both seminal fractions.

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