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PHYSIOLOGICAL, BIOCHEMICAL AND MOLECULAR RESPONSES TO THERMAL
STRESS IN ONGOLE CATTLE
Dr.D.B.V. Ramana, Pr. Scientist (LPM)
Central Research Institute for Dryland Agriculture, Hyderabad-500 059
NATURAL DISASTERS DUE TO CLIMATE CHANGE
CLIMATE CHANGE AND LIVESTOCK
Changes in rainfall amounts and patterns and temperatures
Increase in global average surface temperature between 1.8 and 4.0°C
Approximately 20-30 percent of plant and animal species are expected to be risk of extinction
Resources poor are the most vulnerable and worst affected
Livestock are particularly important for increasing the resilience of vulnerable poor people
Climate change scenarios and projections for India (based on a regional climate modeling system known as PRECIS & IPCC)
Annual mean surface temperature (2.5°C to 5°C) will rise by the end of the century, with warming more pronounced in north India
More than 20% rise in summer monsoon rainfall is projected over all states except Punjab, Rajasthan and Tamil Nadu
overall increase in the intensity of rainfall of between 1 mm and 4 mm/day, except for small areas in north-west India
Range of maximum temperatures in any one season is predicted to vary more widely
Precipitation is expected to increase substantially, particularly over the west coast of India and west central India.
Hydrological cycle is likely to be altered and the severity of droughts and intensity of floods are likely to increase
There will be a general reduction in the quantity of run-off available for agricultural use and drinking water
IMPORTANCE OF LIVESTOCK: GLOBAL AND INDIAN SCENARIO
Livestock systems occupy about 30% of the planet’s ice-free terrestrial surface area
Livestock sector employing approximately 1.3 billion people globally
Livestock provides livelihoods for 600 million smallholding farmers in the developing countries
Livestock production is therefore a key component of world agriculture with multitude of purposes
Indian livestock sector contributes about 6% of the total economy and 25% to agricultural economy
It is estimated that 65% of labor force depends on agriculture and 80% of them are involved in livestock
Region Population
Coastal Andhra 96915
Rayalaseema 83621
Telangana 77125
Total cattle in
Andhra Pardesh
257661
Integrated sample Survey Report-A.P.-2008-10
Ongole cattle status in India
THERMAL STRESS AND IMPACT ON ANIMAL
THERMAL STRESS AND PHYSIOLOGICAL RESPONSES
10
11
12
13
14
15
16
17
18
65
70
75
80
85
90Re
spir
ation
rat
e
THI
40
42
44
46
48
50
52
54
56
58
60
65
70
75
80
85
90
Puls
e ra
te
THI
January February March April May June July August september October November December99.5
100
100.5
101
101.5
102
102.5
103
65
70
75
80
85
90
Rect
al t
empe
ratu
re
THI
THERMAL STRESS AND BIOCHEMICAL RESPONSES
134
135
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144
65
70
75
80
85
90So
dium
(m M
ol/L
)
THI
3
3.2
3.4
3.6
3.8
4
4.2
4.4
65
70
75
80
85
90
Pota
ssiu
m (m
Mol
/L)
THI
January February March April May June July August september October November December91
92
93
94
95
96
97
98
99
100
101
65
70
75
80
85
90
Chlo
ride
(mM
ol/L
)
THI
45
50
55
60
65
70
65
70
75
80
85
90G
luco
se (m
g/dL
)
THI
25
27
29
31
33
35
37
65
70
75
80
85
90
Chol
este
rol (
mg/
dL)
THI
January February March April May June July August september October November December2
2.22.42.62.8
33.23.43.63.8
4
65
70
75
80
85
90
Crea
tine
(mg/
dL)
THI
THERMAL STRESS AND MOLECULAR RESPONSES
HSP27, HSP 70 and HSP 90 expression studies
MC1R gene amplification by using genomic DNA from Ongole cattle
MC1R gene expression
Real time expression analysis reveled that the expression of MC1R gene was relatively upregulated at the culture conditions of 40˚C when compared to control (37˚C) and 25˚C
In Vitro studies
• Heparinized blood samples were processed for lymphocyte isolation using Histopaque
Diluted blood
Histopaque
Pelleted RBCWBC layer
PlasmaCentrifuge 400xg
30 min
Lymphocyte Culture
• WBCs were isolated by centrifugation and washed in RPMI medium
• Live cell count was determined using trypan blue dye• Cell count was adjusted to 5x106/ml • Cells were cultured in RPMI medium containing 10% FCS and
streptomycin, at 450C for 4h.• Cells were processed at 1, 2, and 4h
for RNA and protein isolation.– RNA was isolated using trizol method– Whole cell proteins were extracted
under denaturing conditions
Live cells
Dead cell
0h 1h 2h 4h Ladder HSP 70 (103bp)
HSP70 Expression: At mRNA level
• A gradual increase in mRNA levels was observed during the first two hours followed by a decline at 4th hour
0h 1h 2h 4h Ladder
0h 1h 2h 4h Ladder
HSP 27 (238bp)
HSP 90 (179bp)
Expression of HSP27 and HSP90: At mRNA level
• Expression of both HSP27 and HSP 90 was similar to that of HSP 70
HSP70 Expression: At protein level
Marker 0h 1h 2h 4h 97.466
43
29
18.4
• Increase in the levels of protein followed a similar trend, reaching a maximum at 2 hours followed by a decrease at 4th hour
Acute heat stress (ambient temperature of 40ºC ) induced significant production of ROS, antioxidative enzymes [SOD, catalase (CAT) and lipid peroxidase)
After removal of the heat stress, antioxidative enzymes gradually approached to pre-heat stress exposure levels in a time-dependent manner
Enzymes Control Immediately after exposure to heat
stress for 4 hrs
12h after exposure to heat
stress
SOD (102 U/L) 9.80 ± 1.18 14.60 ± 0.85 9.98 ± 1.21
Catalase (kU/l) 24.30 ± 2.58 40.92 ± 3.24 25.46 ± 1.82
Lipid peroxidase (µM)(TBARS)
0.96 ± 0.06 1.07 ± 0.09 0.97 ± 0.13
In Vivo studies on heat stress recovery In Ongole Bulls
Thank you ….
