A study on IL8RB gene polymorphism as a potential immuno-compromised adherent in exaggeration of parenteral and mammo-crine oxidative stress during mastitis in buffalo

Abstract

The genetic markers in inflammatory responses during mastitis afford a reasonable way for improving milk production in the Egyptian buffalo breed. Among them is the interleukin 8 Receptor Gene (IL8RB) (CXCR2); a chemokine receptor gene augments the neutrophil migration during infection. To understand its role better during mastitis in Egyptian buffalos, twenty-five dairy animals representing the normal, sub-clinically, clinically and chronically affected buffalos were randomly selected from different districts. Screening criteria for mastitis were based on somatic cell count and California mastitis test assays on their milk samples. Biochemically, mastitis induced an increase in milk lactate dehydrogenase, alkaline phosphatase and catalase activities and serum malanoaldehyde concentration. The total antioxidant concentrations, however, decreased in serum and milk during mammary inflammation. The protein profiling of milk whey proved an accelerated mammary inflammatory influx of blood-borne proteins during mastitis. The genomic DNAs were extracted from blood samples and the CXCR2 sequence of 1246 bp covering a part of intron 1, exon 2 and a part of 3'UTR were submitted to Genbank (accession # KY399457.1). The study clearly defined the presence of four SNPs. Three were detected as synonymous substitutions in coding region and one in the 3'UTR region. Only SNP C/A at c.127 was found to be highly associated with mastitis. In conclusion, the results warrant the potential correlation between the genetic SNP variance for certain genes and the incidence of mastitis in buffalo breed.

Keywords: Buffalo; IL8RB; Mastitis; Oxidative stress; Polymorphism; SDS-PAGE.

Graphical abstract

Fig. 1

The protein separation pattern of milk whey using SDS PAGE electrophoresis in normal during mastitis. Lanes 1 is whey milk from control group (10 µL) loading volume. Lane 2 is whey milk from control group (15 µL). Lane 3 is whey milk from mastitic group (10 µL). Lane 4 is whey milk from mastitic group (15µL). The concentration of loaded protein was 1 µg/µL. Lane M is a wide range protein molecular weight marker.

Fig. 2a

Serum activities of lactate dehydrogenase enzyme (LDH) in normal and mastitis samples.

Fig. 2b

Determination of total antioxidant capacity (TAC) in serum in normal and mastitis samples.

Fig. 2c

The serum malondialdehyde (MDA) levels in normal and mastitis samples.

Fig. 2d

Determination of catalase enzyme activity in serum in normal and mastitis samples showing significant elevation in its activity in control samples when compared to other groups.

Fig. 2e

Serum activities of alkaline phosphatase enzyme (AP) in normal and mastitis samples showing significant decrease in its activity in control samples when compared to other groups. The columns with the different letter are significantly differed at P ≤ 0. 01. Two columns with the same letter are not significantly differed at P ≤ 0. 01.

Fig. 3a

LDH activities in control milk and during mastitis show a significant increase in milk LDH activity during mastitis.

Fig. 3b

Determination of TAC in normal milk samples and during mastitis with significant elevation in its value in control milk samples.

Fig. 3c

The levels of MDA in milk in normal samples and during mastitis with observed significant elevation in its level during mastitis.

Fig. 3d

Determination of catalase activities in normal milk samples and during mastitis showing significant elevation in its activity during mastitis.

Fig. 3e

The alkaline phosphatase activities in normal milk samples and during mastitis with doubling of its activity during mastitis. The columns with the different letter are significantly differed at P ≤ 0.01. Two columns with the same letter are not significantly differed at P ≤ 0.01.