EXPRESSION PROFILING, CHARACTERIZATION AND EVALUATION OF HEAT SHOCK PROTEIN 70 (HSP 70) IN VECHUR, KASARGODE (BOS INDICUS) AND CROSSBRED (BOS INDICUS × BOS TAURUS) CATTLE E M Muhammed 1 and T V Aravindakshan 1 Assistant Professor Department of Animal Breeding and Genetics College of Veterinary and Animal Sciences Kerala Veterinary and Animal Sciences University Pookode, Wayanad, Kerala, India M: W:kvasu.ac.in

INTRODUCTION  Global warming, food security and spread of diseases are imminent threats to mankind.  According to the Inter Governmental Panel on Climate Change (IPCC), increase in global average surface temperature by 2100 may be between 1.8 ° C and 4 ° C  Reduction in quality and quantity of animal source food (ASF) may hamper global food security  Susceptible animals may contribute to the spread of contagious as well as zoonotic diseases.  Thus, stress tolerance is a prerequisite for sustainable animal agriculture.

 Vechur is the smallest breed of cattle originated in Kerala state of India  Vechur milk is abundant in beta casein A2, Arginine and have more antibacterial effect than antibiotic ampicillin  Vechur urine possess immunomodulatory property  Vechur cattle are heat tolerant- a property mediated mainly through concerted efforts at physiological, biochemical, cellular and molecular levels  The objective of our study was to validate the role of Vechur DNA in heat tolerance

ANIMAL RESPONSE TO ENVIRONMENTAL STRESS Adaptation Achieved through genetic change (i.e. changes are heritable) over time (generations) and are rarely reversible. Come about via natural selection Acclimation Functional compensation (i.e. physiological or phenotypic change) over a period of days to weeks as in controlled experiments Acclimatisation Functional compensation (i.e. physiological or phenotypic change) over a period of days to weeks as in seasonal climate change

2 YEAR OLD VECHUR AND CROSSBRED COW

30 lactating cows 10 Vechur 10 crossbred 10 Kasargode Physiological, Haematological and biochemical responses Gene and protein expression studies Experimental design 6 Vechur 6 Kasargode 6 crossbred

METEOROLOGICAL DATA  Temperature  Relative humidity  Wind speed  Intensity of solar radiation  Temperature humidity index (THI)  Heat load index (HLI)

RESPONSE VARIABLES Physiological Haematological mRNA Protein Rectal Temperature, Respiratory rate, Pulse rate Total leukocyte count, Differential count, Total RBC count, PCV, ESR, Platelet count, MCV, MCH, MCHC Biochemical Serum cortisol Sequencing-HSP70 and ATP1A1 gene Expression profiling-HSP 70 and ATP1A1 gene SDS PAGE 2D gel electrophoresis

Results

Ta, RH, WS and SR from 8.00 a.m to 2 p.m

THI and HLI from 8.00 a.m to 2 p.m

Relative expression (fold changes) of HSP70 gene after different levels of heat stress within Vechur, Kasargode and crossbred cattle

Relative expression (fold changes) of HSP 70 gene after different levels of heat tress between different genetic groups studied 8.am

Relative expression (fold changes) of ATP1A1 gene after different levels of heat stress within Vechur, Kasargode and crossbred cattle

Relative expression (fold changes) of ATP1A1 gene after different levels of heat tress between different genetic groups studied 8.am

Comparative expression profiles (fold changes) of HSP 70 and ATP1A1 in Vechur, Kasargode and crossbred cattle at different levels of heat stress

Phylogenetic tree constructed based on HSP 70 gene of Vechur cattle with similar sequences in the database

Phylogenetic tree constructed based on ATP1A1 gene of Vechur cattle with similar sequences in the database

Animal model for stress response studies

CONCLUSION  Crossbred animals experienced significant changes in physiological, biochemical, genomic and proteomic expression during exposure to heat stress  Vechur and Kasargode animals experienced similar but less pronounced alterations indicative of superior thermotolerance compared to crossbreds  The enhanced thermal sensitivity of crossbreds indicated that they are reaching at their tolerance limit even at the prevailing climatic conditions  The findings may be applicable in current livestock selection programmes especially in hot and humid climate as well as to plan future strategies in predicted global warming scenario

The heat shock response: Life on the verge of death (Richter et al, Science)

Acknowledgements  Kerala Veterinary and Animal Sciences University, Kerala, India  Department of Animal Husbandry, Kerala, India  National Institute of Animal Nutrition and Physiology, Bangalore, India  University of Western Australia, Perth, Australia  Crawford funding, Australia

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