CSIRO PUBLISHING

Animal Production Science

http://dx.doi.org/10.1071/AN16472

Responses of dairy cows to short-term heat stress in

controlled-climate chambers

J. B. Garner A,B,E, M. Douglas A, S. R. O. Williams A, W. J. Wales A, L. C. Marett A,

K. DiGiacomo B, B. J. Leury B and B. J. Hayes C,D
A
Dairy Production Sciences, Agriculture Research Division, Department of Economic Development, Jobs,
Transport and Resources, 1301 Hazeldean Road, Ellinbank, Vic. 3821, Australia.
B
Faculty of Veterinary and Agricultural Sciences, The University of Melbourne, 142 University Street, Parkville,
Vic. 3053, Australia.
C
BioSciences Research Division, Department of Economic Development, Jobs, Transport and Resources,
AgriBio, 5 Ring Road, Bundoora, Vic. 3083, Australia.
D
Queensland Alliance for Agriculture and Food Innovation, Centre for Animal Science, University of Queensland,
Qld 4072, Australia.
E
Corresponding author. Email: [email protected]

Abstract. The objective of the present research was to describe the physiological and production responses of lactating
dairy cows during and after sudden exposure to temperate-climate heat-wave conditions, compared with cows in
thermoneutral conditions. Twelve lactating multiparous Holstein–Friesian dairy cows were housed in controlled-climate
chambers for 4 days. Six were exposed to a short-term temperature and humidity challenge (THc, diurnal temperature
and humidity fluctuations inducing moderate heat stress; temperature humidity index 74–84) and six cows were exposed
to thermoneutral conditions (THn, temperatur humidity index 55–61). Cows were also measured during a 7-day pre-
experimental and 14-day post-experimental period. Physiological indicators of heat stress were measured, including rectal
and vaginal temperature and respiration rate, which indicated that the THc in controlled-climate chambers induced moderate
heat stress. The cows exposed to the 4-day THc reduced their milk yield by 53% and their dry-matter intake by 48%,
compared with the cows in the THn treatment. Milk yield of THc cows returned to pre-experimental milk yield by Day 7 and
dry-matter intake by Day 4 of the post-experimental period. The short-term heat challenge induced metabolic adaptations
by mobilising adipose tissue, as indicated by increased non-esterified fatty acids, and amino acids from skeletal muscle,
as indicated by increased urea nitrogen to compensate for reduced nutrient intake and increased energy expenditure.
Endocrine responses included greater prolactin concentrations, which is associated with thermoregulation and water
metabolism. The cows exposed to THc displayed production and physical responses that facilitated lower metabolic
heat production and greater heat dissipation in an attempt to maintain homeostasis during the short-term heat exposure.
These results indicated that the conditions imposed on the cows in the controlled-climate chambers were sufficient to
induce heat-stress responses and adversely affected production in the lactating dairy cow, and the delay between the return
to normal feed intake and milk yield following the heat challenge suggests a period of metabolic recovery was occurring.

Additional keywords: endocrine regulation, heat acclimation, moderate heat stress, stress responses, thermal physiology,
thermoregulation.

Received 21 July 2016, accepted 3 February 2017, published online 9 March 2017

Introduction certain thresholds lead to significant declines in feed intake,

Globally, climatic conditions are forecast to become warmer milk production and fertility. These declines are associated
and more variable (IPCC 2014). Consequently, heat stress is with physiological changes, including increased core body
becoming a significant issue for the global dairy industry, as temperature, respiration and panting rates, sweating, metabolic
lactating dairy cows are the most susceptible of domesticated and endocrine-system changes that are important to dairy cow
production animals to the deleterious effects of elevated health and productivity (Collier et al. 1982; Wheelock et al.
temperature and humidity above their thermoneutral zone 2010).
(Kadzere et al. 2002; Bernabucci et al. 2010; Renaudeau et al. Previous research that has formed the basis of our
2012). In dairy cattle, increases in temperature and humidity understanding of heat-stress physiology in the lactating dairy
(often combined as a temperature humidity index, THI) above cow has primarily been conducted on cattle bred in tropical,

Journal compilation
CSIRO 2017 www.publish.csiro.au/journals/an

B Animal Production Science J. B. Garner et al.

subtropical, arid and semiarid climates, which differ significantly winter (June–July 2015) at the Ellinbank Dairy Research
from conditions in temperate climates (Kadzere et al. 2002; Centre, Victoria, Australia.
Renaudeau et al. 2012). Conditions that induce heat stress in
temperate climates are generally characterised by an acute
increase in temperature and humidity ranging from one to Experimental facilities
several days (Ominski et al. 2002). The incidence of sporadic The cows were adapted to the experimental facilities before the
thermal stress that occurs in temperate climates may contribute to commencement of the experiment. The cows were trained in and
significant losses in production and health, as the animals have not acclimatised to the controlled-climate chambers before the
achieved physiological acclimation to chronic high temperature experiment, which involved feeding and handling the cows in
and humidity (Beede and Collier 1986). During short-term heat the controlled-climate chambers for ~5 h a day, 5 days a week for
stress, lactating dairy cows initiate processes to compensate for 4 weeks (with the doors open at ambient conditions). The cows
increased heat load by increasing heat dissipation, reducing feed were also introduced to the experimental diet and outdoor feeding
intake and, subsequently, milk yield between 24 and 48 h after facility for 7 days before the experiment. During the pre- and post-
heat exposure (Gaughan et al. 2009). It has been recognised that experimental periods, the cows were housed in an outdoor,
milk production lost as a result of mild to moderate heat stress is an automatic feed-intake recording facility where they had 24-h
important issue for some temperate climates in the northern access to feed, water and bare paddocks for rest, except during
hemisphere. However, there is very little information describing milking twice daily for ~1.5 h, as described by Garner et al.
the responses of dairy cattle acclimated to an Australian temperate (2016).
climate during short periods of elevated temperature and During the experimental period, the cows were housed
humidity or the subsequent recovery period after exposure. individually in climate-controlled respiration chambers (No
The objective of the present research was to describe the Pollution Industrial Systems, Edinburgh, UK), as described by
production and physiological responses of lactating dairy cattle in Garner et al. (2016).
a temperate climate to moderate, short-term, heat challenge in The conditions in the controlled-climate chambers for the THc
controlled-climate chambers. treatment were designed to remain above THI 74 and not exceed
We hypothesised (1) that sudden exposure to a 4-day heat THI 84, so as to impose a moderate level of heat stress. The
challenge (THc) in controlled-climate chambers will cause climatic conditions programmed into the control system were
declines in milk production and feed intake, and will induce 25C and 60% RH (THI 74) between 1800 hours and 0600 hours,
the physiological indicators of stress, compared with cows in 30C and 50% RH (THI 80) between 0600 hours and 1200 hours,
thermoneutral conditions (THn) in controlled-climate chambers, and 33C and 50% RH (THI 84) between 1200 hours and 1800
and (2) that the cows will display a period of metabolic recovery hours. The THn treatment conditions were programmed at a
following the heat challenge, compared with cows exposed to the constant 14C and 70% RH (THI 59). The cycle of 12 h light
thermoneutral conditions. followed by 12 h dark was controlled manually. The THI was
calculated using the following equation (Yousef 1985):
THI ¼ Tdb þ ð0:36 · TdpÞ þ 41:2; ð1Þ
Materials and methods
where Tdb = hourly dry-bulb temperature (C); Tdp is dew point
Cows and design
temperature (C),
Twelve multiparious non-pregnant Holstein–Friesian cows
(mean
s.d.; 6.4
1.02 years of age, 3.8
1.07 lactations, Tdp ¼ ð237:3 · bÞ=ð1:0  bÞ; b ¼ ½logðRH=100:0Þ
261
24.9 days in milk; 638
34.7 kg bodyweight) were þ ð17:27 · TdbÞ=ð237:3 þ TdbÞ=17:27:
randomly assigned to temperature humidity challenge (THc)
treatment or thermoneutral control (THn) in two groups of six Measurements and sampling
cows, each containing three THc- and three THn-exposed cows. The physiological measurements recorded were respiration
Within each group, cows were then randomly assigned to one of rate (breaths per minute, assessed by counting visible flank
six controlled-climate chambers. The total length of the movements), rectal temperature (212 772 large-animal digital
experiment was 26 consecutive days, with three experimental thermometer; Shoof International, Cambridge, New Zealand),
periods consisting of (1) 7-day pre-experimental period outside in and skin temperature of the flank, neck and udder (AR300+
ambient conditions, (2) 4-day experimental period of THn or THc non-contact infrared thermometer, Smartsensor, Houston, TX,
in controlled-climate chambers and (3) 14-day post-experimental USA). During the pre- and post-experimental periods, respiration
period outside in ambient conditions. During all experimental rate, and rectal and skin temperatures were recorded twice a day
periods, cows were offered a cubed diet ad libitum. The cubes at 0600 hours and 1500 hours, while standing in a race, both on
contained 74% lucerne hay, 25% crushed barley and 1% minerals 2 days per week. The cows were accustomed to standing in this
(calcium, phosphorus and magnesium) and they were formulated race as they usually walk through the race twice daily, on exiting
to meet the cows’ expected requirements for energy, protein and the dairy after milking. During the experimental period in the
minerals. controlled-climate chambers, respiration rate was recorded three
Animal use was approved by the Agricultural Research times daily, at 0600 hours, 1200 hours and 1500 hours, and rectal
and Extension Animal Ethics Committee for the Department and skin temperatures were recorded twice daily at 0600 hours
of Economic Development, Jobs, Transport and Resources, and 1500 hours. The area of skin where the measurement
Victoria, Australia. The experiment was conducted during was taken was marked on each cow with spray paint, so the
Chamber induced heat stress and its effects on dairy cattle Animal Production Science C

measurement was taken from the same place each time. Intra- glucose. Plasma NEFA concentrations were measured using a
vaginal blank CIDRs (Zoetis, Melbourne, Vic., Australia) were kit assay (Wako C NEFA kit, Novachem, Melbourne, Vic.,
modified to house temperature-recording buttons (DS1922 L Australia), as modified by Johnson and Peters (1993). Plasma
iButton; Thermodata, Warrnambool, Vic., Australia) and were glucose concentrations were measured as described by Trinder
inserted into the vagina of each cow for the duration of the (1969) by using the Infinity glucose oxidase liquid stable
experiment. Intra-vaginal temperature was recorded every reagent (Thermo Scientific, Melbourne, Vic., Australia) as per
30-min during the pre- and post-experimental periods and the manufacturer’s instructions. Plasma urea concentrations were
every 15 min during the experimental period. measured using the Infinity urea liquid stable reagent (Thermo
While cows were in the controlled-climate chambers, feed Scientific) as per the manufacturer’s instructions. Plasma analysis
offered and refused were recorded twice daily. Samples of feed for b-hydroxybutyrate was conducted using a b-hydroxybutyrate
offered and refused were collected twice daily at 0600 hours calorimetric assay kit (Cayman Chemical Co., Ann Arbor, MI,
and 1500 hours, for dry matter (DM) and analysis of chemical USA) as per the manufacturer’s instructions. The absorbance of
composition. Samples were processed and analysed for DM, all metabolites was measured on a Thermo Multiskanner plate
crude protein, neutral detergent fibre, acid detergent fibre, reader (Vantaa, Finland). The intra-assay coefficient of variation
lignin, ash, starch and estimated metabolisable energy according for the NEFA, BHBA, urea nitrogen and glucose were 4.1%,
to methods described by Garner et al. (2016). The nutritive 12%, 11.9% and 1.4% respectively.
characteristics of the lucerne cubes with crushed barley grain Blood plasma was analysed for concentrations of insulin
was DM 94.7%, crude protein 18.1% DM, neutral detergent (double-antibody radioimmunoassay, Tindal et al. 1978),
fibre 39.2% DM, acid detergent fibre 30.7% DM, lignin 6.7% growth hormone (double-antibody radioimmunoassay Downing
DM, ash 7.7% DM, starch 6.8% DM and estimated metabolisable et al. 1995), insulin-like growth factor 1 (double-antibody
energy 9.9 (MJ/kg). radioimmunoassay with human recombinant IGF-I, ARM4050,
Cows were milked twice daily at 0600 hours and 1500 hours, Amersham-Pharmacia Biotech, Buckinghamshire, England, and
with yields recorded automatically in the milking parlour at antihuman IGF-I antiserum, AFP4892898, Torrance, CA, USA,
each milking during the pre- and post-experimental periods following acid-ethanol extraction and cryoprecipitation, Breier
(MM25; DeLaval International, Tumba, Sweden). Whereas in et al. 1991), cortisol (radioimmunoassay using ImmuchemTM
the controlled-climate chambers, the cows were milked using Coated Tube Cortisol 1251 RIA kits, Biomedicals, Asse-
an inbuilt milking system (same clusters and pulsators as the Relegem, Belgium), leptin (double-antibody : radioimmunoassay,
milking parlour), and milk yields were recorded manually. Blache et al. 2000) and prolactin (homologous double antibody
Samples for milk-composition analyses were taken from six RIA using standard NIADDK-oPrl-l2 and antiserum R160,
milkings (3 · AM and 3 · PM) each week during pre- and Miller et al. 1995). The intra-assay coefficients of variation for
post-experimental periods, and from each milking during the the insulin, growth hormone, IGF-1, cortisol, leptin and prolactin
experimental period. Samples were analysed for fat, protein were 4.6%, 7.9%, 4.8%, 5.1%, 6.5% and 9.7% respectively.
and lactose by a near-infrared milk analyser (Model 2000,
Bentley Instruments, Chaska, MN, USA). Energy-corrected Statistical analyses
milk (ECM) was calculated using the equation of Tyrrell and
All statistical analyses were performed using ANOVA in
Reid (1965):
GENSTAT (17th edition; VSN International, Hemel Hempstead,
ECM ðkg=cow:dayÞ UK). Data were analysed for differences of means within
experimental period between the THc and THn treatment
milk yield ðkgÞ · ð376 · fat% þ 209 · protein% þ 948Þ
¼ : groups. The pre-experimental values were fitted as the covariate
3138 for the analyses. Significance was declared when P < 0.05 and
Water intake was recorded automatically in the controlled- trends were declared when P < 0.10, unless otherwise indicated.
climate chambers by using calibrated flow meters in the water Results are presented as mean values and pooled standard error
troughs. Bodyweights were recorded daily during the pre- and of the mean.
post-experimental periods using walk-over scales. Body
condition score was assessed using an 8-point scale (Earle Results
1976) once during the pre- and post-experimental periods.
During the experimental period, excrement was collected and Production measurements
weighed twice daily (0600 hours and 1500 hours), using a faecal Dry-matter intake (DMI) was not different among the treatment
chute and urine was kept separate using a urine separator. groups during the pre-experimental period. Mean DMI during the
Blood samples were taken by coccygeal venipuncture at 1600 experimental period was lower (P < 0.001) for the THc-exposed
hours into two vacutainers containing lithium heparin and EDTA, cows than the cows exposed to THn, with the THc-exposed cows
once on Day 5 of the pre-experimental period, once on Day 4 of having lower feed intake than the THn-exposed cows on Days 2,
experimental period and once on Day 11 of the post-experimental 3 and 4 of the heat challenge (Table 1, Fig. 1a), and, on Day 4, this
period. Blood samples were placed immediately on ice and decline in feed intake was 48% lower than for the THn-exposed
centrifuged within 15 min of collection at 531g for 15 min at cows. There was a trend (P < 0.06; Table 1, Fig. 1a) for the
4C, and plasma was removed and stored at 20C until analysis. THc-exposed cows to have a lower DMI during the post-
Plasma was analysed for concentrations of non-esterified fatty experimental period than that of the THn-exposed cows. The
acids (NEFA), b-hydroxybutyrate (BHBA), urea nitrogen and digestibility of DM was not different among treatment groups.
D Animal Production Science J. B. Garner et al.
Table 1. Production parameters of the thermoneutral control (THn) and temperature humidity challenge (THc) cows during the pre-experimental, experimental and post-experimental periods
ECM, energy-corrected milk. P-values are between treatments and within period. Data presented are treatment-group means with pooled standard error of the mean (s.e.m.). Trends are identified at *P < 0.10, and

s.e.m.
Post-experimental (14 days)

0.31
0.56

0.31
0.08
0.07
0.08
0.02
0.03
3.36
However, the faeces DM % was lower for the THc-exposed cows

–
–
–
–
1
during the experimental period (Table 1). The intake of water was
lower (P < 0.05) for the THc-exposed cows than for the THn-

17.4**

5.58
4.04
4.92
1.00
0.69
0.84
23.5*
THc

18.7
exposed cows on Day 4 of the experimental period. There was a

–
–
–
–
663
trend (P < 0.10) for the THc-exposed cows to consume a smaller
proportion (33.5%) of their total daily DM during the day of the

5.44
3.96
4.81
1.10
0.68
0.85
final day of the treatment in the controlled-climate chambers than
THn
24.4
18.1
21.1

–
–
–
–
667
the proportion (42.5%) consumed by THn-exposed cows. During
the post-experimental period, the THc-exposed cows returned
s.e.m.

1.35
1.03

0.46
0.12
0.09
0.06
0.03
0.12
to their pre-experimental DMI by Day 4 and the THn-exposed
1.1

5.0
0.9
3.0
7.0
–
cows returned to pre-experimental measurements by Day 3
(Fig. 1a).
0.53***
0.36***
12.6***

10.6***
7.9***

4.32**

0.52**
Day 4
THc

72.6**
5.3**
Milk yield of the THn- and THc-exposed cows was not
6.31

4.86

–
65.9

21.7
different during the pre-experimental period. The THc-exposed
cows had a lower milk and ECM yield during the experimental
5.63
3.87
4.96
1.02
0.71
1.02

period (P < 0.001) than did the THn-exposed cows (Table 1,

THn
24.2

23.2

66.3

19.6
92.4
7.9
–
17

Fig. 1b). By Day 4 of the treatment, the milk and ECM yields of
the THc cows were 53% and 54% lower than those of the THn-
s.e.m.
1.65
1.16
1.68

0.28
0.12
0.09
0.07
0.03
0.04

5.8
0.6
1.9
4.9
–

exposed cows (Table 1). The mean percentage of milk protein was
higher for the THc-exposed cows than the THn cows during
statistical difference is identified at **P < 0.05, and ***P < 0.001

the experimental period on Days 3 (P < 0.05) and 4 (P < 0.05).

10.6***
9.7***

4.23**

0.61**
0.44**
0.51**
Day 3

13.1**

5.1**
THc

5.80

4.83

However, the yield of protein was lower for the THc-exposed

52.4

21.1
78.3

cows during the experimental period than that for the THn-
exposed cows on Days 3 (P < 0.001) and 4 (P < 0.001;
5.10
3.81
4.89
0.89
0.66
0.86
THn
21.6
17.6
20.8

62.9

21.3
87.2
7.9
–

Table 1). The mean yield of fat was lower for the THc-
exposed cows than for the THn-exposed cows during the
experimental period on Days 3 (P < 0.05) and 4 (P < 0.001),
s.e.m.
1.32
1.68
2.36

0.49
0.13
0.11
0.13
0.07
0.10

2.0
0.5
1.6
5.6
–

as was the mean yield of lactose during the experimental period

on Days 3 (P < 0.001) and 4 (P < 0.05; Table 1). Once cows were
16.4**

14.2**

6.1**
Day 2
THc

4.40
1.14
5.33
0.61
0.55
0.74

returned to ambient conditions during the post-experimental

13.9

62.7

24.4
79.2
–

period, milk yield was lower (P < 0.05) for the THc-exposed
cows than the THn-exposed cows (Table 1, Fig. 1b), with the
4.80
3.94
5.16
0.89
0.71
0.96
THn
21.9
18.1
21.9

64.8

20.5
78.5
7.6
–

THc cows returning to their pre-experimental milk yield by Day 7

and the THn cows returning to pre-experimental measurements
s.e.m.

by Day 3 (Fig. 1b).

0.98
1.15

0.50
0.12
0.12
0.12
0.05
0.07
1.8

1.8
0.5
2.1
5.2
–

22.9**

Physiological measurements
Day 1

4.70
4.20
5.30
0.76
0.70
0.80
THc
20.3
16.5

63.6

85.1
7.4
–
18*

The vaginal and rectal temperatures of the THc-exposed cows

were higher (P < 0.001) than those of the THn-exposed cows
5.80
3.90
5.10
1.05
0.70
0.90

during each day of the experimental period (Fig. 2a, b, Table 2).
THn
21.5
17.7
23.1

61.5

16.6
76.7
7.8
–

The flank, neck and udder temperatures of the THc-exposed

cows were higher during each day of the experimental period
s.e.m.
1.54
1.28
1.04

0.41
0.14
0.13
0.05
0.04
0.06
Pre-experimental (7days)

17.7

than were those of the THn-exposed cows (Table 2). The

–
–
–
–

mean respiration rate of the THc-exposed cows was higher on

Treatment days 1 (P < 0.05), 3 (P < 0.05) and 4 (P < 0.10) than was
4.81
3.91
4.82
0.85
0.71
0.87
THc
22.6
18.2

–
–
–
–
664
21

that of the THn-exposed cows (Table 2).

The pre-experimental concentrations of NEFA (mmol/L),
4.57
3.82
4.82
0.82
0.69
0.88

glucose (mmol/L), plasma urea nitrogen (mmol/L) and BHBA

THn
22.3
18.2
19.9

–
–
–
–
667

(mmol/L), insulin (mU/mL), cortisol (ng/mL), leptin (ng/mL), GH

(ng/mL), IGF-1 (ng/mL) and prolactin (ng/mL) were not different
Daily water intake (L/day)

between THc and THn groups (Table 3). The THc-exposed cows
Daily faeces (kg DM/day)

on Day 4 of the heat challenge had an 83% increase (P < 0.05)

Daily urine (kg/day)
DM Intake (kg/day)
Milk yield (kg/day)

in plasma NEFA concentration compared with the THn-exposed

Milk composition

cows. There was a trend (P < 0.10) for the THc-exposed cows to
Lactose (kg)
Lactose (%)
ECM (kg/day)

Protein (kg)
Protein (%)

have a 42% increase in plasma urea concentration, 675% increase

Bodyweight
Fat (kg)
Fat (%)
Parameter

in plasma prolactin concentration and, a 24% reduction in plasma

DMD%

insulin concentration compared with the THn-exposed cows on

Day 4 of the heat challenge.
Chamber induced heat stress and its effects on dairy cattle Animal Production Science E

30 (a)

25

20

DMI (kg)
15
Heat challenge

10 Thermoneutral

Baseline Challenge Recovery

5

25 (b)

20
Milk yield (kg/day)

15

Heat challenge
10
Thermoneutral

Baseline Challenge Recovery

5
0 2 4 6 8 10 12 14 16 18 20 22 24 26
Experiment day

Fig. 1. (a) Dry-matter intake (DMI, kg) and (b) milk yield (kg) per experimental day with standard error
of the mean error bars. Experimental Days 1–4 coloured in blue for thermoneutral control (THn) and red
for temperature humidity challenge (THc) cows.

Controlled-climate chambers and ambient conditions cows on Day 4), and a proportion of this heat stress-induced
The cows in the THn treatment remained in thermoneutral decline in milk yield may be explained by the 48% reduction
conditions, ranging from 10.2C to 15.9C and from 61.5% to in feed intake (11.6 kg DM/day less than for the THn cows on
94.9% RH (THI = 55–61, Fig. 2a). The cows in the THc treatment Day 4). This is supported by pair-feeding experiments that
experienced cyclical daily temperatures and relative humidity (to identified that only a portion (35–50%) of the heat stress-
mimic diurnal patterns), ranging from 21.3C to 32.8C and from induced milk-yield decline can be explained by reduced
35.8% to 88.9% RH (THI = 69–83; Fig. 2b). During the pre- nutrient intake (Rhoads et al. 2009; Wheelock et al. 2010).
and post-experimental periods, the cows experienced ambient Wheelock et al. (2010) and Rhoads et al. (2009) measured a
weather conditions, ranging from daily averages of 4.1C to feed-intake reduction of 30% and 35% respectively, and a milk-
13.2C (mean 8.7C) and 66.7% to 99% RH (mean 89.1%), yield decline of 27.6% and 40% respectively, during heat stress;
with THI of 47–58 (mean 52.5). however, the pair-fed controls in both experiments were
consuming the same feed intake and they reduced their milk
Discussion yield by only 13.9% and 21% respectively. The remaining
proportion of loss in milk production may be a consequence
Production responses of changes in nutrient partitioning and thermoregulation. Heat-
This investigation demonstrated that sudden-onset short-term, stressed animals are known to have increased energy expenditure
moderate heat stress, similar to that which occurs in temperate for heat loss via panting and sweating (Fuquay 1981). It is
climates, can negatively affect production during and after estimated that maintenance energy requirements may increase
exposure in lactating dairy cows. Reductions in feed intake by up to 25% during heat stress to compensate for the energy-
and milk production are well known responses to heat stress expensive processes of thermoregulation (NRC 1989). In support
that lower metabolic heat production. In support of our first of the second hypothesis, we speculate, that following the TH
hypothesis, the THc-exposed cows by Day 4 of the experimental challenge, the cows experienced a period of metabolic recovery
period reduced their milk yield by 53% (9.1 L/day less than THn as there was a delay between the return to pre-experimental feed
F Animal Production Science J. B. Garner et al.

39.5 65
(a)
THn vaginal temperature
THn THI

39.0 60

Controlled-climate chambers THI

Vaginal temperature (°C)

38.5 55

41.0 (b) THc vaginal temperature 85

THc THI

40.5
80

40.0

75
39.5

39.0 70
9: 3: 9: 3: 9: 3: 9: 3: 9: 3: 9: 3: 9: 3: 9: 3:
30 30 30 30 30 30 30 30 30 30 30 30 30 30 30 30
:0 :0 :0 : :0 : :0 : : : :0 : :0 : :0 :
0 0 0 00 0 00 0 00 00 00 0 00 0 00 0 00
AM PM PM AM AM PM PM AM AM PM PM AM AM PM PM AM

Fig. 2. (a) Vaginal temperature of thermoneutral control (THn) cows and the temperature humidity
index (THI) of the controlled-climate chambers, and (b) vaginal temperature of the temperature
humidity challenge (THc) cows and the THI of the controlled-climate chambers during the 4-day
experimental period.

intake occurring on Day 4 and the return to baseline milk yield by night. This is in agreement with changes in feeding behaviour
Day 7 of the post-experimental period. previously reported from heat-stressed dairy cows (Schneider
The short-term heat exposure caused considerable reductions et al. 1988). This feeding pattern has been investigated as an
in milk production. The THc-exposed cows produced a 48% amelioration strategy to reduce the metabolic heat associated with
lower milk fat and 49% lower lactose and protein yield on Day 4 of fermentation during the hottest parts of the day (Ominski et al.
the treatment period than did the THn-exposed cows. This is 2002; Aharoni et al. 2005). The reduction in total DMI, and the
consistent with previous research that identified that heat-stressed shift to consume the majority of feed at night is thought to be a
cows secrete ~200–400 g less milk lactose than do pair-fed behavioural adaptation that reduces metabolic heat production
thermoneutral control cows (Wheelock et al. 2010; Baumgard by lowering heat of fermentation in the rumen, and thereby
et al. 2011). The amount of secreted lactose is associated with the reducing core body temperature.
amount of available glucose secreted from the liver and, as this
glucose is utilised at an increased rate by extramammary tissues, Physiological indicators of heat stress
the available glucose is preferentially utilised for processes other
The physiological indicators of stress measured during the
than milk secretion in the heat stressed cow (Baumgard and
experiment indicated that the cows in the THc treatment were
Rhoads 2013). The reduction in milk-fat yield of the THc
experiencing hyperthermia, which supports our first hypothesis.
cows is also consistent with another experiment reporting a
The diurnal THI range (69–83) imposed on the cows in the THc
reduction of 39% in the milk fat yield of Holstein cows
treatment was consistent with a mild to moderate heat-stress
exposed to 30C (McDowell et al. 1976). The decline in milk
THI range (Renaudeau et al. 2012). The body temperatures
fat during heat stress could be attributed to the reduced energy
were consistently higher in the THc cows than the THn cows,
intake (Collier 1985).
which indicated that the cows in the heat challenge were unable
to maintain normal body temperature and were suffering from a
Feeding pattern heat-stressed state, results of which are consistent with those of
In response to the TH challenge, cows altered their feed-intake previous studies under similar conditions (Rhoads et al. 2009;
pattern. On the final day of the experimental period, there was a Wheelock et al. 2010). We chose to mimic typical summer
trend that indicated that the THc-exposed cows consumed a lower conditions in southern Australia and, although the temperature
percentage of their total daily DM intake during the day, dropped to 25C for 12 h overnight, the THc cows still exhibited
preferring to consume the majority during the cooler period at elevated core body temperature and respiration rates on the
Chamber induced heat stress and its effects on dairy cattle Animal Production Science G

During baseline, cows in both groups were treated identically (housed outside in ambient conditions and allowed to eat ad libitum). Treatment Days 1–4 in controlled-climate chambers, cows were either

s.e.m.
exposed to the THc or the THn and were allowed to eat ad libitum. During recovery, cows in both groups were treated identically (housed outside in ambient conditions and allowed to eat ad libitum). P-values
are between treatments and within period. Data presented are treatment group means with pooled standard error of the mean (s.e.m.). Trends are identified at *P < 0.10, and statistical difference is identified
Physiological parameters of the thermoneutral control (THn) and temperature humidity challenge (THc) cows during the pre-experimental, experimentalal and post-

0.09
0.09

1.22
0.66
0.70
2.17
morning of each treatment day, which indicated that the heat

Post-experimental
stress was being maintained for the entire 24 h of each

(14 days)
treatment day. The vaginal temperature of the THc-exposed cows

THc
38.2
38.7

25.3
28.5
33.4
increased each day of the 4-day heat challenge, indicating

43
an accumulation of body heat as the elevated THI during the
night did not allow for sufficient heat dissipation to occur. The

THn
38.4
38.6

25.8
28.1
34.6
45
increase in respiration rate observed in the THc-exposed cows
is characteristic of heat-stressed cows attempting to maintain

s.e.m.
0.13
0.17
5.59

1.06

0.83
thermoregulation by increasing evaporative heat loss from the

0.8
respiratory-tract surface (Silanikove 2000). The THc cows also
had greater skin temperatures for each area of the body measured
40.4***
40.7***

39.1***
40.4***
41.3***
Day 4

at each time point during the treatment period, which indicated

THc

86*
that the THc cows were dissipating heat through their skin
surface, most likely due to increased vasodilation enabling the
transfer of heat from blood to the periphery (Choshniak et al.
THn
38.6
39.1

30.6
30.4
35.7
54

1982; Silanikove 2000). However, a proportion of this increase

in skin surface temperature will be attributed to the higher
s.e.m.

ambient temperature inside of the chambers for the heat-

0.14
0.17
5.58

1.32
0.98
1.11
challenged treatment. The elevated THI challenge in this
experiment initiated the characteristic physiological indicators
40.4***
40.6***

38.5***

of stress and heat-dissipation mechanisms.

Day 3

88.8**

36.9**

39.2**
THc

Metabolic and endocrine responses

THn
38.5
38.9

31.3
32.1
34.9

The THc-exposed cows displayed physiological changes,

56
at **P < 0.05, and ***P < 0.001

suggesting that adipose tissue was being mobilised to

experimental periods

Skin temperature (C)

compensate for reduced energy intake, as indicated by elevated

s.e.m.
0.15
0.13
5.17

1.28
0.82
0.82

concentrations of circulating NEFA (Bauman and Currie 1980).

Increased plasma NEFA concentrations in response to the TH
challenge may be associated with decreased skeletal-muscle
39.8***
40.1***

38.9***
39.6***
Day 2

37.6**
THc

glucose uptake, glucose oxidation and hepatic gluconeogenesis

74

(Randle 1998). However, others have reported that cows

exposed to prolonged heat-stress conditions do not mobilise
THn
38.5
38.9

31.2
32.1
35.2

adipose tissue to compensate for reduced energy intake, even

52

though they are in an negative energetic state and markedly lose

bodyweight, which has been attributed to the antilipolytic action
s.e.m.
0.13
0.05
5.13

1.06
0.72
0.88

of heat-stress increased basal insulin concentrations (Rhoads

et al. 2009; Shwartz et al. 2009). We speculate that, as there
was no heat stress-induced increase in insulin (THc cows had
39.3***

36.8***
38.1***
39.1***
Day 1

38.5**
80.6**
THc

24% lower plasma insulin than did the THn cows), adipose tissue
was being mobilised without being inhibited by the antilipolytic
actions of increased insulin concentration. The short-term heat
THn
38.5
38.2

29.3
30.9
34.9

exposure of the cows (4 days) may not have been sufficient time
42

for major shifts in metabolism or loss in bodyweight to occur.

In addition to adipose tissue, there was a trend suggesting
s.e.m.

7.32
0.04
0.09

0.72
0.36

that skeletal-muscle protein was being mobilised during heat

1.0
Pre-experimental

stress to support energy requirements. Plasma urea N (PUN) can

(7days)

initiate from three sources; amino acids from the diet, or a waste
THc
38.3
38.9

25.8
29.2
35.3

product of rumen ammonia formation into microbial protein

43

or hepatic deamination of amino acids from skeletal muscle

(Wheelock et al. 2010). The 43% increase in PUN of the cows
THn
38.2
38.8

26.6
28.9
33.1
43

in response to the THc indicated that amino acids were being

mobilised from protein sources in addition to adipose tissue.
RR (breaths/minute)

This increase in PUN is in agreement with Shwartz et al. (2009)

who measured a 47% increase from baseline PUN in response
Vaginal (C)

to 7 days of heat stress in lactating Holsteins.

Rectal ( C)
Parameter
Table 2.

In response to the heat exposure, there was a trend suggesting



Udder
Flank

an increase in plasma prolactin in the THc cows. Prolactin is

Neck

known to increase with rapid changes in air temperature and has

H Animal Production Science J. B. Garner et al.

Table 3. Concentrations of plasma metabolites and hormones for the thermoneutral control (THn) and temperature humidity challenge (THc)
cows on day 5 of the pre-experiment period, day 4 of the experimental period and day 11 of the post-experimental period
NEFA, non-esterified fatty acids. P-values are between treatments and within period. Data presented are treatment group means with pooled standard error of
the mean (s.e.m.). Trends are identified at *P < 0.10, and statistical difference is identified at **P < 0.05, and ***P < 0.001

Variable Pre-experimental Experimental Post-experimental

THn THc s.e.m. THn THc s.e.m. THn THc s.e.m.
NEFA (mM/L) 0.09 0.08 0.02 0.06 0.11** 0.01 0.07 0.07 0.01
Glucose (mmol/L) 4.47 4.27 0.13 4.29 4.49 0.11 4.21 4.45 0.18
Urea (mmol/L) 6.31 4.86 0.53 4.87 6.93* 0.79 6.06 6.26 0.55
BHB mM/L 0.36 0.37 0.04 0.45 0.45 0.02 0.43 0.47 0.03
Insulin (mU/mL) 15.0 15.1 2.18 26.4 20.1* 2.49 14.1 12.8 1.38
Cortisol (ng/mL) 4.71 3.00 1.93 3.70 3.22 0.93 4.68 3.41 0.85
GH (ng/mL) 2.08 2.83 0.33 3.06 3.47 0.55 2.34 2.44 0.23
Leptin (ng/mL) 0.25 0.23 0.07 0.35 0.29 0.06 0.42 0.41 0.09
IGF-1 (ng/mL) 33.5 28.7 5.35 33.1 29.3 3.18 40.0 39.3 2.80
Prolactin (ng/mL) 1.00 0.38 0.29 0.54 4.19* 1.52 1.12 1.09 0.55

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