Fluorometric Determination of Avian Sperm Viability and Concentration

Bilgili, S. F.; Renden, J. A.

doi:10.3382/ps.0632275

Cited by 117 publications

(36 citation statements)

References 10 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…The progressive decline in the spermatozoa concentration was recorded with the increase in frequency of collection which is in agreement to the previous studies on poultry birds (Santayana 1985) and turkey (Zahraddeen et al 2005). The acrosomal and plasma membrane integrity of Indian Red Jungle Fowl spermatozoa remained similar (P>0.05) in all collection frequencies according to previous studies in other poultry birds (Bilgili and Renden 1984;Bilgili et al 1985;Donoghue et al 1995;. It is the same for the proportion of morphologically normal sperm.…”

Section: Discussionsupporting

confidence: 79%

Semen characteristics of the Indian Red Jungle Fowl (Gallus gallus murghi)

et al. 2015

View full text Add to dashboard Cite

The Indian Red Jungle Fowl is a wild native gallus subspecies of Southern Asia. Semen has never been studied in this species. In order to better know the male reproductive capacities, experiments were conducted to study the semen characteristics, impact of ejaculate collection frequencies, and timing of collection on sperm quality parameters. Mean sperm concentration 800 million/mL, total sperm per ejaculate (0.015 billion), motility (63.5 %), live/total sperm (92.4 %), intact acrosome (75.5 %), and plasma membrane integrity (89.2 %) were recorded. Percentage of abnormal sperm (head, mid-piece, and tail) was 8.1 % and recovered mainly mid-piece abnormalities. The motile sperm percentage was positively correlated with intact acrosomes (r=0.34) and plasma membrane integrity (r=0.41). Total sperm per ejaculate (billion) was maximum at 72 h of collection followed by 24 and 48 h of collection. Daily and weekly sperm production (billion) was found maximum at 24 h of collection compared to 12, 48, and 72 h of collection. Sperm motility was higher at 24, 48, and 72 h of collection compared to 12 h of collection, but the number of live sperm were higher at 12 h of collection compared to 24, 48, and 72 h. Sperm concentration was better in the morning time, while the values for sperm viability and plasma membrane integrity were higher in the semen collected at evening time. In conclusion, the Indian Red Jungle Fowl shows a semen production quantitatively relatively low for the species as compared to domestic chicken and contrasted parameters of quality. The semen production is affected by the frequency of collection with an optimum for a daily collection preferentially held in the evening period. These results may now be used for artificial insemination and conservation program.

show abstract

Section: Discussionsupporting

confidence: 79%

Semen characteristics of the Indian Red Jungle Fowl (Gallus gallus murghi)

et al. 2015

View full text Add to dashboard Cite

show abstract

“…Initial studies of PM integrity using fluorescent probes in spermatozoa, used ethidium bromide (BILGILI;RENDEN, 1984;HALANGK et al, 1984), which exhibits DNA specificity and only penetrates cells with damaged PM. EB fluorescence increases when it intercalates into the double helix (PAL et al, 1998).…”

Section: Pm Integritymentioning

confidence: 99%

Techniques for sperm evaluation using fluorescent probes

2015

View full text Add to dashboard Cite

A variety of laboratory tests were developed to obtain more reliable results of sperm evaluation and increase the accuracy of sperm fertility predictions. These tests detected damage of sperm specific compartments or organelles, which cannot be detected in routine sperm analysis. The use of fluorescent probes and detection using fluorescent microscopy or flow cytometry is an important tool but a more precise and accurate laboratory test is needed. Propidium iodide and 6-carboxyfluorescein diacetate are used for evaluations of plasmatic membrane integrity. Fluorescein isothiocyanate, associated with conjugated lecithin Psium sativum or Arachis hypogaea, are used for evaluations of acrosome integrity. Two probes, MitoTracker or Rhodamine123, are generally used to measure the absence or presence of mitochondrial potential. However, a better option is 5,5'; 6,6' -tetrachloro -1,1'; 3,3' -tetraetilbenzimidazolil-carbocyanine (JC-1) dye, which assesses not only the presence of mitochondrial potential and distinguished spermatozoa with poorly and highly functional mitochondria. Two techniques, TUNEL or COMETA, and the Acridine Orange Test (AOT) dye are used to evaluate chromatin integrity. A fluorescence technique based on chlortetracycline (CTC) or Merocyanine 540 is used to estimate whether sperm pass by or through the capacitation process. This review focuses on the fluorescent probes that are most widely used to evaluate plasma membrane integrity, capacitation, acrosome integrity, chromatin integrity and mitochondrial potential. Key words: Fluorescence microscopy, sperm, cryopreservation ResumoDiversos testes laboratoriais têm sido desenvolvidos com o intuito de obter resultados confiáveis nas avaliações espermáticas, aumentando a qualidade e a confiabilidade do sêmen utilizado em técnicas de reprodução assistida. Esses testes têm permitido detectar danos em compartimentos e organelas específicas da célula espermática, que não são detectados nas análises de rotina. Dentre esses testes, o uso de sondas fluorescentes e sua detecção em microscópio de epifluorescência ou citometria de fluxo se tornaram ferramenta importante quando uma avaliação mais acurada é necessária. Para a avaliação da integridade de membrana plasmática, pode ser utilizado o iodeto de propídio, associado ao diacetato de 6-carboxifluoresceína. As avaliações de integridade acrossomal podem ser feitas através do isotiocianato de fluoresceína, associado às lecitinas conjugadas, como a Psium sativum ou Arachis hypogaea. O potencial mitocondrial pode ser avaliado quanto à ausência ou presença através da sonda Mitotracker ou Rodamina 123. Outra opção ainda melhor pode ser observada utilizando o corante iodeto de 5,5'; 6,6' -tetracloro -1,1'; 3,3' -tetraetilbenzimidazolil-carbocianina (JC-1), este corante avalia a presença do potencial mitocondrial e avalia quanto à classificação do grau. Para avaliar a integridade

show abstract

“…The semen to be used in the afternoon was stored at 4°C and occasionally agitated. Sperm concentration (Cecil, 1982) and viability (Bugili & Renden, 1984) was determined immediately before each artificial insemination (AI) period. For AI, the tip of a micropipettor (Gilson Pipetman, Rainin Instru.…”

Section: Introductionmentioning

confidence: 99%