Measuring haemolysis in cattle serum by direct UV–VIS and RGB digital image-based methods

Abstract

A simple, rapid procedure is required for the routine detection and quantification of haemolysis, one of the main sources of unreliable results in serum analysis. In this study, we compared two different approaches for the rapid determination of haemolysis in cattle serum. The first consisted of estimating haemolysis via a simple direct ultraviolet–visible (UV–VIS) spectrophotometric measurement of serum samples. The second involved analysis of red, green, blue (RGB) colour data extracted from digital images of serum samples and relating the haemoglobin (Hb) content by means of both univariate (R, G, B and intensity separately) and multivariate calibrations (R, G, B and intensity jointly) using partial least squares regression and artificial neural networks. The direct UV–VIS analysis and RGB-multivariate analysis using neural network methods were both appropriate for evaluating haemolysis in serum cattle samples. The procedures displayed good accuracy (mean recoveries of 100.7 and 102.1%, respectively), adequate precision (with coefficients of variation from 0.21 to 2.68%), limit of detection (0.14 and 0.21 g L–1, respectively), and linearity of up to 10 g L–1.

Assessment of Babesia spp. prevalence in various domestic animals across Southern Punjab, Pakistan

“…Genomic DNA was extracted from randomly selected samples using the Wiz prep® Genomic DNA purification kit. A set of oligonucleotide primers, GAU9 (5'-CTGTCGTACCGTTGGTTGAC-3') and GAU10 (Reverse) (3'-CGCACGGACGGAGACCGA-5'), targeting a 541 bp amplicon of the Babesia 18S rRNA gene, was used as described by (Larrán et al, 2022). The PCR reaction mixture (25 µl) contained 1X Taq Buffer, 0.2 mM dNTPs, 1.5 mM MgCl2, 2.5 U/l Taq polymerase (Fermentas, UK), 4 mM of each primer, 2 µl of DNA template, and DNasefree deionized water.…”

“…DNA amplification was performed using a thermal cycler (Gene Amp® PCR system 2700 Applied Biosystems Inc., UK). The thermo-profile described by (Larrán et al, 2022) was modified for the current study, with an initial denaturation at 94°C for 10 minutes, followed by 40 cycles of denaturation at 94°C for 30 seconds, annealing at 62°C for 1 minute, and elongation at 72°C for 1 minute. The final extension was done at 72°C for 5 minutes.…”