Effect of In Ovo Injection of L-Arginine in Different Chicken Embryonic Development Stages on Post-Hatchability, Immune Response, and Myo-D and Myogenin Proteins

SIMPLE SUMMARY: In the current study, we hypothesized that the in ovo injection of L-arginine (L-Arg) at different stages of embryonic development, which would have positive effects on the survival rate, hatching rate, immunoglobulin M (IgM) levels, heat shock proteins (HSPs) such as HSP-47, HSP-60, and HSP-70, and muscle development markers as well: Mainly, myoblast determination protein (myoD) and myogenin in pectoral muscles. As indicated, the in ovo injection of L-Arg resulted in an increased hatch rate and weight, survival rate, higher levels of IgM, and myogenin and MyoD expression in the muscles. At the same time, a decrease in the level of HSP-47, HSP-60, and HSP-70 expressions in the tissues was observed on the 14th day of injection compared to the eighth and 18th day of the injection period. In addition, the in ovo injection of L-Arg decreased the serum glutamate oxaloacetate transaminase (SGOT) and serum glutamate pyruvate transaminase (SGPT) concentration in serum as well micronuclei and nuclear abnormality in the blood on the 14th day of the incubation period. Hence, the 14th day would be a suitable day for the injection of L-Arg to promote the hatching rate and muscle growth of broiler chickens. ABSTRACT: The aim of this study was to evaluate the effect of in ovo injection with different ratios of L-arginine (L-Arg) into Ross broiler eggs at three different embryonic developmental stages (eighth day (d), 14th day, and 18th day) on the survival, hatchability, and body weight (BW) of one-day-old hatched chicks. Additionally, we have analyzed the levels of serum glutamate oxaloacetate transaminase (SGOT) and serum glutamate pyruvate transaminase (SGPT), the protein expression of heat shock proteins (HSPs), and we have also determined micronuclei (MN) and nuclear abnormality (NA). In addition, the genotoxic effect was observed in peripheral blood cells such as the presence of micronuclei and nuclear abnormalities in the experimental groups. The results showed that survival and hatching rates as well as body weight were increased on the 14th day of incubation compared to the eighth and 18th day of incubation at lower concentrations of L-Arg. Moreover, the levels of SGOT and SGPT were also significantly (p < 0.05) increased on the 14th day of incubation at the same concentration (100 μg/μL/egg) of injection. In addition, immunoglobulin (IgM) levels were increased on the 14th day of incubation compared to other days. The protein expressions of HSP-47, HSP-60, and HSP-70 in the liver were significantly down-regulated, whereas the expression of myogenin and myoblast determination protein (MyoD) were significantly up-regulated on the 14th day after incubation when treated with all different doses such as 100 μg, 1000 μg, and 2500 μg/μL/egg, namely 3T1, 3T2, and 3T3, respectively. However, the treatment with low doses of L-Arg down-regulated the expression levels of those proteins on the 14th day of incubation. Histopathology of the liver by hematoxylin and eosin (H&E) staining showed that the majority of liver damage, specifically intracytoplasmic vacuoles, were observed in the 3T1, 3T2, and 3T3 groups. The minimum dose of 100 μg/mL/egg on the 14th day of incubation significantly prevented intracytoplasmic vacuole damages. These results demonstrate that in ovo administration of L-Arg at (100 μg/μL/egg) may be an effective method to increase chick BW, hatch rate, muscle growth-related proteins, and promote the immune response through increasing IgM on the 14th day of the incubation period.