Research Article
Volume 5 Issue 6 - 2020
The Effect of Ergot Alkaloid Consumption on Bull Sperm Chromatin Structure and Acrosome Integrity
AN DeCarlo1, CR Burnett1, P Rowan1, R Johnston1, RE Ricks1, WC Bridges Jr2 and Scott L Pratt1*
1Animal and Veterinary Sciences Department, Clemson University, Clemson, South Carolina, USA
2Mathematical Sciences Department, Clemson University, Clemson, South Carolina, USA
*Corresponding Author: Scott L Pratt, Animal and Veterinary Sciences Department, Clemson University, Clemson, South Carolina, USA.
Received: April 20, 2020; Published: May 20, 2020




Abstract

Bull sperm chromatin and acrosomal integrity when consuming ergot alkaloids (EA) was assessed. Bulls consumed rations containing or lacking EA and were subjected to semen collection every 21 to 28 days beginning at the start of feeding. Sperm samples, both fixed in formalin and post-thaw were analyzed. To determine chromatin damage, a variation of sperm chromatin structure assay (SCSA) was utilized. This technique was validated by exposing ~1 X 106 formalin fixed sperm to UV light at 254 nm, 302 nm or 365 nm for 0, 30, 60 and 90 minutes, in triplicate, to induce chromatin damage prior to staining. Sperm membranes of fixed and post-thaw samples were permeabilized with a 0.1% triton solution, then incubated with a staining buffer containing 6 mg/ml Acridine Orange stain solution and pelleted. Cells were resuspended in Agilent cell chip buffer and analyzed with the Agilent Cell Chip Assay. To evaluate acrosomal integrity, sperm samples from all years were incubated in the presence or absence of PNA-Alexa Fluor 594 (1 mg/ml) and counterstained with DAPI. Smears were produced and sperm were counted from five randomly chosen fields from each slide using a 40x objective on an Axio Imager 2 fluorescence microscope. Sperm were evaluated as percent stained out of total cell number. The SCSA was effective using the Agilent Bioanalyzer Cell Chip Assay; however, EA treatment exhibited no effect on chromatin structure. Sperm labeled with PNA-Alexa Fluor 594 showed 4 staining patterns, and EA altered these patterns, indicating exposure impacts acrosomal integrity.

Keywords: Acridine Orange; Acrosome; Chromatin; Peanut Agglutinin; Semen

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Citation: Scott L Pratt., et al. “The Effect of Ergot Alkaloid Consumption on Bull Sperm Chromatin Structure and Acrosome Integrity”.EC Veterinary Science 5.6 (2020): 12-21.

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