Studies on Liquefaction Time and Proteins Involved in the Improvement of Seminal Characteristics in Dromedary Camels (<i>Camelus dromedarius</i>)

In most of domestic animals, semen biology is well studied, but it is not so well studied in camelids particularly in dromedary camels. In camelids, the ejaculated semen is highly viscous, so before its evaluation and processing for preservation, it is necessary to be liquefied. Our study was designed to evaluate the effect of three different enzymes (0.1% Collagenase, 0.5 X Accutase, 0.1% Trypsin-EDTA - Ethylene-diamine-tetra-acetic acid) on seminal viscosity at different time intervals along with its effects on seminal parameters. Semen was collected from the six adult healthy male camels using female camel as dummy. Bovine artificial vagina was used for the semen collection and ejaculate was kept at 37ºC for further processing after its collection. All three enzymes were diluted in 1x phosphate buffered saline (PBS) and the semen samples were kept at 37ºC. The samples were evaluated at different time intervals 0, 5, 10, 15, 30, 60, 120 and 240 min (T0–T240) for viscosity and it was evaluated by pipette method. Immediately after collection, the viscosity was very high, and no sperm mass motility was seen. However, after enzyme treatment viscosity was showed declining trend in all treatment groups compared to control. In trypsin treated group maximum motility was reached in 15 minutes. Live-dead percent was observed to be similar among treatment groups. Observations on liquefaction of semen revealed that use of enzymes decreasing the viscosity but with different rates.

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“…In a previous study, complete liquefaction was observed at 23.89±1.49H varying from 18 to 41H without any enzyme treatment (Mal et al, 2016).…”

Section: Discussionmentioning

confidence: 84%

Effect of different proteolytic enzymes on liquefaction of semen of dromedary camel

et al. 2018

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“…Algunos investigadores aplicaron licuefacción mecánica, agitación 5 , agitación 12,13 , punción y pipeteo 14 y centrifugación 15 , últimamente, exposición a ultrasonido con éxito para eliminar viscosidad y mejorar la preservación enfriada a corto plazo del semen de dromedario 11 . Otros autores complementaron el diluyente de semen con diferentes enzimas a fin de eliminar viscosidad, uso de catalizadores de descomposición con peróxido de hidrógeno 16 , catalizadores de descomposición de carbohidratos de cadena larga 17,18 , enzimas proteolíticas 14,15,[19][20][21][22] . Avances recientes del análisis de semen asistido por computadora (ASAC) demostraron su capacidad para una estimación precisa de función espermática y heterogeneidad en comparación con la evaluación tradicional del semen 23 .…”

Section: Introductionunclassified

Parámetros cinéticos de espermatozoides en semen fresco y crioconservado de alpaca (Vicugna pacosL.)

Apaza-Callisaya¹,

Loza-Murguia²,

Quispe-Paxipati³

et al. 2020

J. Selva Andina Anim. Sci.

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La crioconservación de los epz de alpaca es importante para la mejora genética. El presente trabajo tuvo como propósito evaluar los parámetros cinéticos de spz en semen fresco y crioconservado en dilutores, mediante el equipo ASAC. Se utilizaron 3 alpacas machos, que fueron entrenados durante un mes en la colección de semen con la vagina artificial. El esperma se congeló empleando dos diluyentes (AndroMed comercial) y (AndroMed comercial, ácido cítrico, gentamicina y glicerol). El semen tuvo las siguientes características, volumen 1.5±0.7 mL, viscosidad 5.9±1.5, pH de 7.1, con tendencia a alcalinidad y el color que predominó fue el blanco translucido. Los promedios de parámetros cinéticos en semen fresco fueron VCL (25.8±4.2 µm/s), VSL (10.3±2.5 µm/s) y VAP (18.9±3.2 µm/s). Los parámetros cinéticos en semen crioconservado fuero el dilutor 2 VCL (8.4±1.4 µm/s), el dilutor 1 (7.2±1.2 µm/s), VSL el dilutor 2 (3.4±0.8 µm/s), dilutor 1 (2.9±0.7µm/s), VAP dilutor 2 (5.8±1.0 µm/s), dilutor 1 (4.8±0.8 µm/s). Se concluye que el dilutor 2 constituye una alternativa más para la crioconservación de semen de alpaca.

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“…Elimination of semen viscosity is one of the major topics that gained considerable attention of camelidae reproduction researchers over the past recent years (Rateb et al., 2020). Among other postulations, presence of proteins in the seminal plasma has been suggested to contribute to camelides semen viscosity (Kershaw‐Young et al., 2013; Mal et al., 2016). In the present results, non‐purified (T 0 ) protease‐liquefied semen showed a marked improvement in post‐thaw sperm motility compared to that of control.…”

Section: Discussionmentioning

confidence: 99%

“…Enzymatic liquefaction of camel semen, particularly by using proteolytic enzymes, has been considered the most effective for elimination of camel semen viscosity and improving motility and kinematics of spermatozoa (Kershaw‐Young et al., 2017; Mal et al., 2016; Monaco et al., 2016). Nevertheless, protease‐based semen liquefaction was recently reported to drastically affect sperm vitality, acrosome integrity and sperm cell surface glycocalyx pattern especially after exposure to the stress of freezing/thawing cycle (El‐Bahrawy et al., 2017; Rateb et al., 2019).…”

Section: Introductionmentioning

confidence: 99%

Purification of cryopreserved camel spermatozoa following protease‐based semen liquefaction by lectin‐functionalized DNA‐defrag magnetic nanoparticles

Rateb

2020

Reprod Domestic Animals

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Although incorporating proteases into sperm medium is considered the most effective procedure to eliminate camel semen viscosity, it drastically affects viability, morpho‐functional properties and, hence, fertilization potential of spermatozoa. The present work aimed at evaluating adequacy of employing magnetic nanoparticles‐based sperm purification technique for eluting impaired and apoptotic camel spermatozoa from cryopreserved semen doses following protease‐based semen liquefaction. Thirty cryopreserved semen doses (50 x 106 sperm/straw) representing the following liquefaction treatments: control (untreated), 0.1 mg/ml papain or 5 U/ml bromelain were used (n = 10 straws per treatment). Immediately after thawing (38°C for 40 s), sperm concentration of each straw within treatment was readjusted to 15 x 106 sperm/mL by dilution in PBS (37°C). Sperm physical and cytological properties were then assessed (non‐purified semen). Thereafter, each specimen was subjected to lectin‐functionalized DNA‐defrag magnetic nanoparticles sperm purification, and the same sperm traits were re‐evaluated after undergoing purification (purified semen). Sperm DNA fragmentation level within each group, prior to and after magnetic nano‐purification, was also determined by fluorescent imaging. The results showed a dramatic improvement (p < .05) in post‐thaw motility (%), viability (%), normal sperm (%), intact acrosome (%) and HOST‐reacted (%) spermatozoa in protease‐liquefied semen following sperm magnetic nano‐purification. Additionally, the highest (p < .05) DNA fragmentation level was recorded in all cryopreserved semen groups prior to purification, whereas the lowest (p < .05) was observed in the protease‐treated specimens after magnetic nano‐purification. These results indicate that protease‐based semen liquefaction prior to cryopreservation in conjunction with magnetic nano‐purification post‐thawing holds potential for reducing the proportion of damaged and dead spermatozoa, hence improving camel sperm fertilization competence.

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Studies on Liquefaction Time and Proteins Involved in the Improvement of Seminal Characteristics in Dromedary Camels (Camelus dromedarius)

Cited by 13 publications

References 24 publications

Effect of different proteolytic enzymes on liquefaction of semen of dromedary camel

Effect of different proteolytic enzymes on liquefaction of semen of dromedary camel

Parámetros cinéticos de espermatozoides en semen fresco y crioconservado de alpaca (Vicugna pacosL.)

Purification of cryopreserved camel spermatozoa following protease‐based semen liquefaction by lectin‐functionalized DNA‐defrag magnetic nanoparticles

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