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
damarla97@gmail.com
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
Ongole cattle status in India
Region
Population
Coastal Andhra
96915
Rayalaseema
83621
Telangana
77125
Total
cattle
in
257661
Andhra Pardesh
Integrated sample Survey Report-A.P.-2008-10
THERMAL STRESS AND IMPACT ON ANIMAL
THERMAL STRESS AND PHYSIOLOGICAL RESPONSES
18
90
85
16
15
80
THI
Respiration rate
17
14
75
13
12
70
11
10
65
60
90
58
56
85
52
80
THI
Pulse rate
54
50
48
75
46
44
70
42
40
65
103
90
85
102
101.5
80
101
75
THI
Rectal temperature
102.5
100.5
70
100
99.5
65
January
February
March
April
May
June
July
August
september
October
November December
THERMAL STRESS AND BIOCHEMICAL RESPONSES
144
90
142
85
141
140
80
THI
Sodium (m Mol/L)
143
139
138
75
137
136
70
134
65
4.4
90
4.2
85
4
3.8
80
3.6
75
THI
Potassium (mMol/L)
135
3.4
70
3.2
3
101
65
90
99
85
98
97
80
THI
Chloride (mMol/L)
100
96
95
75
94
93
70
92
91
65
January
February
March
April
May
June
July
August
september
October
November December
65
85
60
80
55
75
50
70
45
37
65
90
THI
90
35
85
33
80
THI
Glucose (mg/dL)
Cholesterol (mg/dL)
70
31
75
29
70
25
65
4
3.8
3.6
3.4
3.2
3
2.8
2.6
2.4
2.2
2
90
85
80
THI
Creatine (mg/dL)
27
75
70
65
January
February
March
April
May
June
July
August
september
October
November December
THERMAL STRESS AND MOLECULAR RESPONSES
HSP27, HSP 70 and HSP 90 expression studies
MC1R gene expression
MC1R gene amplification by using genomic DNA from Ongole cattle
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
Centrifuge 400xg
30 min
Plasma
WBC layer
Pelleted RBC
Histopaque
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
Dead cell
for RNA and protein isolation.
– RNA was isolated using trizol method
– Whole cell proteins were extracted
under denaturing conditions
Live cells
HSP70 Expression: At mRNA level
0h
1h
2h
4h
Ladder
HSP 70 (103bp)
• A gradual increase in mRNA levels was observed during the
first two hours followed by a decline at 4th hour
Expression of HSP27 and HSP90: At mRNA level
0h
1h
2h
4h
Ladder
HSP 27 (238bp)
0h
1h
2h
4h
Ladder
HSP 90 (179bp)
• Expression of both HSP27 and HSP 90 was similar to that of HSP 70
HSP70 Expression: At protein level
97.4
66
Marker
0h
1h
2h
4h
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
In Vivo studies on heat stress recovery In Ongole Bulls
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
12h after
exposure to heat exposure to heat
stress for 4 hrs
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
Thank you ….