66 DEVELOPMENTAL HEALTH ASSESSMENT OF OFFSPRING PRODUCED FROM MAGNETIC NANOPARTICLES USING NEAR INFRARED ANALYSIS OF PLASMA

Abstract

In vitro fertilization is optimized when there is a homogenous population of viable spermatozoa, not subjected toxic waste products of apoptotic cells. In a previous study, we developed a “nanopurification” technique to magnetically target and remove non-viable spermatozoa from a boar insemination dose. Nanopurified semen has successfully been studied with IVF in swine and bovine but lacks health data regarding offspring produced from exposed semen. Developmental health performance in mammals is typically assessed through measurements of immune related biomolecules in plasma (e.g. immunoglobulins), quantifying each variable with a specific analytical assay. Recent developments in aqueous based near infrared spectroscopy (NIR), aquaphotomics, have been shown to distinguish reproductive stages (e.g. oestrus, diestrus) in blood serum. Thus, application of aquaphotomics may be ideal for analysis of offspring resulting from fertilization with nanopurified semen, using serum or plasma. Our study objective was to identify holistic differences in blood plasma by characterising NIR spectral profiles in offspring produced from nanopurified semen. Extended boar semen doses were mixed with or without specific nanoparticles to target non-viable spermatozoa. Semen doses were exposed to an electromagnetic field, noninvasively separating non-viable spermatozoa from the insemination dose. Six gilts were bred with (n = 3) or without (n = 3) nanopurified semen. Following birth and weaning, 20 offspring of equal sexes were randomly selected from control and nanopurified litters (10/group) for growth and developmental measurements up until market weight. Blood plasma was collected from offspring at market weight for NIR analysis. Spectral data were collected with a quartz cuvette and ASD FieldSpec^® 3 spectrophotometer (ASD Inc., Boulder, CO, USA). Chemometric analysis (Unscrambler^® X version 10.4; CAMO Software, Oslo, Norway) included a Savistsky-Golay 1st and 2nd derivative for detection of distinct spectral features. Principal component analysis and partial least-squares block-discrimination were used to examine treatment effects, in a blind experiment. Plasma spectral profiles from control and nanopurified offspring contained 6 shared peaks at 1360, 1373, 1402, 1404, 1422, and 1428 nm. Principle components 1 and 2 accounted for 96.26% of the total variance, with no separation of principal component analysis scores for plasma spectra between groups. Partial least-squares discriminant analysis metrics (slope = 0.026, SECV = 0.52) and Students t-test showed no significant difference (P = 0.57) between groups. Results indicate blood plasma content is not influenced in nanopurified offspring when compared with the control. In addition, solute NIR has shown to be a valuable promising tool for assessing complex aqueous solutions in swine. Further effects on growth and development from offspring born from nanopurified continue to be investigated.

This work was supported by USDA-ARS Biophotonics Initiative grant #58–6402–3-018.