Lung function measurements using spirometry in small cetaceans:
Tursiops truncatus and Phocoena phocoena
S.Gans
3274683
12-07-2013
Project Tutor University of Utrecht
dr. M.G. van Emst
Project Tutor Dolfinarium
C.E. van Elk DVM
Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
phocoena
Table of contents
Abstract…………………………………………………………………………………………………………….…………….3
Used abbreviations………………………………………………………………………………………………………….4
Introduction…………………………………………………………………………………………………………………….5 - 6
Aim of the study………………………………………………………………………………………………………………7
Materials and methods………………………………………………………………………………………………..…8 - 14
Animals……………………………………………………………………………………………………………………….…..8
Harbour porpoises…………………………………………………………………………………….….…….8
Bottlenose dolphins………………………………………………………………………….………….……..9
Equipment……………………………………………………………………………………………….……………..……….9
Measurements…………………………………………………………………………………………………………………9
Parameters …………………………………………………………………………………………………….…..10
Preparing the spirograph…………………………………………………………………………….………10
Harbour porpoises………………………………………………………………………….…………………..10
Bottlenose dolphins……………………………………………………………………..……………..……..10
Sick or healthy………………………………………………………………………………………………………...………11
Processing data…………………………………………………………………………………………………….………...11
Statistics…………………………………………………………………………………………………………………..……..14
Results………………………………………………………………………………………………………………………...15 - 41
Harbour porpoises……………………………………………………………………………………………………..……15
Overview results……………………………………………………………………………………………..….15
Flow/volume curves…………………………………………………………………………………………...15
Statistics and graphs…………………………………………………………………………………………...18
Correlation……………………………………………………………………………………………..18
Regression………………………………………………………………………………………………19
Inside versus outside the water………………………………………………………………25
Animals with confirmed pulmonary disorder versus healthy animals……..26
Bottlenose dolphins…………………………………………………………………………………………………………30
Overview results………………………………………………………………………………………………...30
Flow/volume curves……………………………………………………………………………………………30
Statistics and graphs………………………………………………………………………………….……….33
Correlation …………………………………………………………………………….……..………33
Inside versus outside the water …………………………………………………………….34
Animals with confirmed pulmonary disorder versus healthy animals…….37
Discussion………………………………………………………………………………………………………….………..42 - 45
Harbour porpoises…………………………………………………………………………………………………...........43
Bottlenose dolphins……………………………………………………………………………………………………..….44
Acknowledgements……………………………………………………………………………………………..….……...46
References……………………………………………………………………………………………………………..……….47
Attachments………………………………………………………………………………………………………...….…..48-86
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Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
phocoena
Abstract
Pulmonary function testing, such as spirometry is a validated method to evaluate lung
mechanics and resistance of the conducting airways. Until this moment there is very limited
experience in the use of spirometry in small cetaceans.
In this study 13 harbour porpoises (Phocoena phocoena) and 7 bottlenose dolphins (Tursiops
truncatus) were measured using a pneumotachograph specifically designed for this purpose.
They were measured inside as well as outside the water. Three of the animals had confirmed
respiratory disorders. The measurements were well tolerated and the results were
reproducible and reliable. The results represented a good overview of spirometric data in
small cetaceans. Correlations were found in the porpoises between body size and
spirometric data. Furthermore, differences were found in bottlenose dolphins while inside
or outside the water, and in both species in animals with confirmed lung disorders. Because
of the limited number of animals further studies are needed to confirm these findings.
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Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
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Used abbreviations
VC: Vital Capacity
TLC: Total Lung Capacity
TV: Tidal Volume
PEF: Peak Expiratory Flow
PIF: Peak Inspiratory Flow
Te: Expiration time
Ti: Inspiration time
Vins: Inspiration Volume
Vex: Expiration Volume
Df: Degrees of freedom
SD: Standard Deviation
df: degrees of freedom
SEM: Standard Error of the Mean
CT: Computed Tomography
β: regression coeficent
ρ: population correlation coefficient
P: calculated probability
t: test result
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Introduction
Like many species, cetaceans are perfectly adapted to their particular niche. Because of their
life in the water behavioral, physiological and anatomical features are different from
terrestrial animals. An extremely important part of the life of a cetacean is diving. 1, 2
The large toothed whales, such as sperm whales (Physeter macrocephalus) typically dive
approximately 400-800 meters for a duration of 40-60 minutes. Bottlenose dolphins on the
other hand, often dive for less than 5 minutes to shallow depths. The maximum depth
measured is 390 meters. Harbour porpoises dive to depths of approximately 14-40 meters. 3
To make diving possible there are marked changes in the delivery, storing and utilizing of
oxygen compared to terrestrial animals. 2, 4
It seems logical to assume a cetacean would have large lungs to carry much oxygen. This is
not the case, as large lungs that are filled with air would cost the animal a lot of energy to
overcome the large buoyancy during a dive. 1, 4 The lung volume of diving mammals is
therefore dictated by body size. 5 As marine mammals don’t have large lungs for the storage
of oxygen, they have to get their oxygen elsewhere when diving. Oxygen is transported and
stored in the body by reversibly binding to myoglobin found in the muscle, or to hemoglobin
found in the red blood cells. 2, 3, 6 7 Marine mammals store oxygen in their muscles by
increased myoglobin stores 2, 3, 7, the concentration of myoglobin is 10-30 times higher than
found in terrestrial animals. 3 They also possess more hemoglobin, because of an increased
blood volume 7 and also because they have a higher concentration of hemoglobin. 3, 7 The
amount of oxygen stored in the blood of small cetaceans ranges from normal to 3 times the
amount found in terrestrial animals.
As there is a limited amount of oxygen available during a dive, marine mammals have to be
careful how to spend it. They have developed mechanisms to preserve their storage of
oxygen for as long as possible. The first adaptation is a decrease in heart rate during dives.
Normal heart rates during a dive are 30-50 beats per minute. When ascending after a dive
the animals are tachycardic, with heart rates between 120-150 beats per minute. This
maximizes respiratory gas exchange and cardiac output and refills the muscles and blood
with oxygen for the next dive. The second adaptation is peripheral vasoconstriction; blood
flow is conserved for the brain and the heart while other organs receive almost no blood. 3, 7
Marine mammals can tolerate low concentrations of oxygen in the blood and have
adaptations in their brains for better hypoxemic tolerance. Their anaerobic metabolism is
much more developed. 3
The airways in cetaceans are reinforced with cartilage from the trachea to the level of the
alveolar sac. When the lung collapses, air is forced out of the alveoli into the reinforced
bronchi. 3, 5, 7 This prevents both air trapping and gas exchange. Stopping gas exchange is
very important for the prevention of decompression sickness. 3, 5 The reinforcements in the
airways also contribute to the fast inhale and exhale of large volumes of air. 5, 7
The blowhole located on the top of the heads of cetaceans is the most obvious adaptation of
their respiratory system for life in the water. The position of the blowhole allows the animals
to breath quickly with little to no interruption during locomotion. Cetaceans do not have
conchae. The lack of conchae contributes to the fast inhale and exhale of large volumes of
air. 2 Inside the entrance of the blowhole the nasal plug is located. This plug occludes the
airway when in neutral position and stops water from entering the respiratory system, even
at high pressures. When the animal breathes muscles contract, pulling the plug forward and
opening the airway. 8
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Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
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The larynx of cetaceans is the most specialized of all marine mammals. It has the form of an
elongated goose beak and separates the digestive tract form the respiratory tract. 9 Because
of this adaptation cetaceans are obligate nasal breathers. 2
Respiratory tract infections are an important factor in the mortality and morbidity in
dolphins. 10 Dolphins do not show symptoms of a disease until a late stage because they are
prey animals. 11 When they have a respiratory tract infection the symptoms are not specific.
Dolphins cough mainly when they have a problem in the upper airways. A higher breathing
frequency is only seen if a substantial part of the lung is damaged. 11 Often the only
symptoms are a lower appetite or anorexia. 11 12 It is very important to recognize an infection
at an early stage, because treatment is more difficult in a more advanced stage of disease. 13
Auscultation is very difficult in small cetaceans, because of their thick layer of blubber, their
fast breathing and the loud noises the animals produce. Blood work can show a leucocytosis
and left warded shift, but this test is not reliable because of the large variation in results
between animals. At this moment plasma fibrinogen is the most used indicator of an
infection in cetaceans. It can be used to identify an infection at an early stage. Plasma
fibrinogen is however not always reliable and it is not specific for the identification of a
respiratory infection. 11 CT-scanning is very useful in diagnosing an infection in the lungs.
However, using this technique is complicated, expensive, and the animals have to be taken
out of the water and transported to the scanner, which is very stressful.
Sputum culture can be a useful diagnostic tool and could deliver relevant information, but
often commensal bacteria are found. It is therefore very difficult to determine if the grown
micro-organism is the cause of the infection. 11 Bronchoscopy can be used to circumvent this
problem. A diagnosis can be made using a bronchial washing or by using a protected brush.
Bronchoscopy is however an invasive method and, just like using a CT-scan, the animals have
to be taken out of the water. 14, 15
Diagnosing a problem in the respiratory tract of dolphins is difficult and often invasive. A
non-invasive diagnostic technique that can be used to recognise a problem in the respiratory
tract of dolphins would therefore be extremely helpful. Spirometry is theoretically an ideal
tool that can be used to make a diagnosis, as it might be possible to document the integrity
of the respiratory system in morphological and physiological sense. Spirometry can be used
for both screening as for management of disease. As for now there is very limited experience
in the use of spirometry in small cetaceans.
In the Dolfinarium (marine mammal park located in the Nederlands) research has been
performed for about a decade trying to improve the quality of diagnosing and managing
respiratory disease in small cetaceans. 16 In these studies advanced techniques are used
applying techniques commonly used in human medicine. Apart from spirometry, endoscopy
and CT-scanning have been evaluated.
This research is funded by the Dolfinarium and by private contributions.
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Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
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Aim of the study
This study has several aims.
 The determination of spirometric parameters of healthy bottlenose dolphins and
porpoises.
 The determination of inter-individual and intra-individual variation of the
measurements, both with animals in and out of the water.
 Comparing and describing the spirometric parameters of healthy bottlenose dolphins
and porpoises.
 Performing measurements in diseased animals in order to determine which
parameter(s) might be useful to recognise pathology. Most of the time these will be
stranded animals.
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Materials and methods
Animals:
Harbour porpoises (Phocoena phocoena)
13 porpoises were measured, which were kept at two different locations (table 1 and 2). The
first is the Bruinvisbaai, here the healthy animals are located that cannot be returned into
the wild because of various reasons.
The second location is SOS Dolfijn where stranded animals are located for rehabilitation and
return to the wild.
Name
Sex
Age in months
Length in cm
Average weight
in kg
Amber
Berend
Ellen
Ester
Gerhard
Joelle
Jose
Siepy
Female
Male
Female
Female
Male
Female
Female
Female
108 (or older)
22
93
11,5
23
11
36
71
154
118
149
116
116
127
136
147
45,1
29,4
50,7
36,7
34
36,9
40,1
44
Table 1: animals measured at Bruinvisbaai, data arranged alphabetically
Name
Sex
Age in months
Length in cm
Average weight
in kg
Desiree
Renske
Sietske
Thom
Tonia
Female
Female
Female
Male
Female
58
70 (or older)
22
12
22
149
159
119
105
122
50,8
72
29,3
22,7
31,9
Table 2: animals measured at SOS Dolfijn, data arranged alphabetically
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Bottlenose dolphins (Tursiops truncatus)
7 bottlenose dolphins were measured. (table 3 and 4) These animals were also located on
two different locations. The first location is the Domein. The animals that perform in a show
for general public are housed here.
The second location is the Sickbay. Sick animals are kept separate from the rest of the
animals in this location. The length of the single female dolphin was unknown, so her length
was estimated based on the data of the other animals.
Name
Sex
Age in months
Length in cm
Weight in kg
Apollo
Juno
Nemo
Tlisaka
Tsalka
Tucker
Male
Male
Male
Male
Male
Male
292
352
319
148
184
388
285
274
262
275
274
289
225
208
223
217
217
232
Table 3: animals measured at the Domein, data arranged alphabetically
Name
Sex
Age in months
Length in cm
Weight in kg
Maaike*
Female
360
**
266
Table 4: animal measured in the Sickbay
*Maaike was pregnant in the first trimester when the measurement was performed.
**Due to her location in the sick bay it was not possible to measure this animal without
taking her out of the water, which was considered inappropriate just for this purpose.
Equipment:
A Masterscope (Viasys HC) modified spirometer manufactured by CareFusion using an
adapted pneumotachograph with conforming software and hardware. The sample frequency
of the pneumotachograph is 0,5 kiloHerz, Resistance= 36 Pascal/Liter/seconds.
A blowhole-mask has been designed and manufactured also especially for this study. The
mask fits both the porpoises and the dolphins. It is manufactured out of silicone and doesn’t
irritate the skin, does not leak air when put over the blowhole and can be disinfected. It is
connected to the pneumotachograph by a hose of eight centimetres diameter.
Measurements
The study was performed in the Dolfinarium from 02-11-2012 to 30-05-2013.The
measurements were performed on both animal species while in the water as well as outside.
The animals were only measured outside the water when the animals were taken out of the
water for other reasons. The animals were not taken out of the water specifically for this
experiment, therefore minimising stress and avoiding discomfort of the animal. As many as
possible representative and repeatable measurements were performed in every animal.
Length and weight of the animals were recorded. The breathing effort had to be
incorporated in the interpretation of the results. Therefore observations were made to
distinguish between a maximum effort breath and a tidal volume breath. Obviously this is a
subjective and qualitative observation. In diseased animals the spirometric results were
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Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
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correlated with disease activity as measured by routinely used methods. The gathered data
were evaluated immediately on technical quality and reliability, and were saved for further
analysis. Afterwards they were converted in workable tables and graphs.
A dierexperimentencommissie (DEC) appliance was not considered necessary by the
responsible veterinarian as performing spirometry is non-invasive and does not cause any
discomfort to the animals.
Parameters
1.
2.
3.
4.
5.
Vital features: species, weight, length, age, sex
Absence or presence of disease
Inside or outside of the water
Volume parameters: vital capacity, inspiratory capacity, expiratory capacity
Flow parameters: expiration time, inspiration time, expiratory to inspiratory ratio,
maximum expiratory and inspiratory flow
6. Further observations: shape and features of the flow/volume curves, subjective
judgement of the breathing technique
7. Integration of 1-6 including determination of averages, standard deviations, intraindividual and inter-individual variation
Preparing the spirograph
Before each measurement the measuring equipment was prepared and calibrated with a
three liter volume pump. The pneumotachograph needed fifteen minutes to reach working
temperature. The current ambient conditions were then added into the program. After this
the calibration of the spirograph was checked once more. This calibration measurement was
also used to calculate the correction factor. The use of a correction factor is necessary
because of the large, adapted pneumotachograph. The software has been designed for the
normal sized pneumotachograph for use in humans.
Measuring the animals
The trainers helped the animals to go on shore or remain steady in the water. The mask was
placed over the blowhole and was fit tightly to prevent air leakage. This was especially
important in the smaller animals. Measurements were performed by allowing the animal to
breathe a couple of times (mostly 1-4 times) with the mask on. Then the mask was removed,
so the animal could breathe freely preventing dead space ventilation as much as possible.
The mask was then placed back on the animal so more breathing efforts could be measured.
This was continued until a sufficient number of breathing efforts was recorded. As many
breathing cycles as were feasible under the circumstances were recorded. In between
different animals the mask and the hose were disinfected with alcohol.
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Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
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Sick or healthy
The animals were placed in four categories.
0. No signs or symptoms of disease
1. Most likely no pulmonary disorder
2. Most likely pulmonary disorder
3. Definite pulmonary disorder
In order to make this judgement a number of diagnostic tools was used.
1. Behaviour of the animals
2. Blood tests
3. Bronchoscopy
4. CT-scans
5. Necropsy
Processing data
The data were processed using Microsoft excel for mac 2011. When processing a series of
data the file containing the data of the calibration pump was processed first. Using this data
the correction factor was calculated. (see graph 1) The correction factor in this series of
measurements is 12, as a 3 liter volume pump was used. Graph 1 shows a volume of 0,25
liter which is 1/12th of three liter. Because of the calibration before the measurements this
value is the same in all measurements.
Graph 1: Flow volume curves of the calibration pump
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Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
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The timeframe in which the animal breathes was determined by flow versus time and
volume versus time curves. This facilitates searching for the breathing in the raw data. Graph
2 shows an animal exhaling approximately 30 seconds after the start of the measurement.
After this the animal inhales and exhales 4 times.
Graph 2 Flow (l/s) and Volume (l) versus Time (s), measurement of harbour porpoise (Thom)
Flow/volume curves were then constructed. Because of the sensitivity of the machine the
wind and small air vibrations are also measured, this causes the volume to shift during the
measurements. Sometimes the animal exhales more air than it inhales or vice versa. This
also affects the volume, as it does not return to zero after that breathing session. These
effects can be seen in graph 2. Because of these effects the volume had to be adjusted to
zero by hand before each breathing session. Then all flow/volume curves were plotted into
one graph according to international standards of the European Respiratory Society/
American Thoracic Society.
In some porpoises the flow is very low at the end of an inspiration. It is therefore difficult to
determine when the inspiration stops. Strictly spoken it ends when the flow reaches zero,
but because these are difficult measurements and the equipment is very sensitive, a fixed
value as the end of the inspiration was determined. As the mean flow of the porpoises is
around 7,5 l/s, 3 1/3 % is 0,25. Flow was considered zero when decreasing below this
threshold.
From the flow/volume curves the vital capacity (VC) and the volume of each inspiration or
expiration (Vins, Vex) could be determined. It was decided to only include the volumes of
the inspirations or expirations that were no less than 50% of the vital capacity. The other
breathing efforts were considered too small, this was most likely due to a reaction of the
animal to the mask. The mean and standard deviation (SD) of the Vins and Vex were
calculated.
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The peak expiration flow (PEF) and peak inspiration flow (PIF) could be determined with the
help of the graph or from the raw data. The shape of the flow/volume curve was decisive in
considering if a measurement was representative. If for example the curve has a very high
flow but a low volume compared to the other curves the flow value was not used.
The expiration and inspiration time (Te, Ti) could be determined from the raw data. The
mean and SD of the Te and Ti were calculated. The ratio of Te and Ti was also calculated.
Only the closed flow/volume curves were used for this, because the animal than exhales and
inhales the same amount of air. A curve was determined closed if the difference in
inspiration and expiration volumes was less than 10%. The mean and SD of the Te/Ti ratio
were calculated.
Measurements of obviously poor quality by any reason were disregarded.
Breathing cycles were excluded when the shape of their flow/volume curve was clearly
abnormal. In these measurements the animals inhaled or exhaled in two parts as shown in
graph 3. Incomplete breathing cycles were also rejected, for example only an exhalation.
Graph 3: Flow/Volume curve of a bottlenose dolphin (Tsalka)
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Statistics
SPSS statistics for mac version 21 and the SAS system were used to perform the statistics.
Correlation analysis and linear regression were performed on the data. The animals with
confirmed pulmonary disease were disregarded. Measurements performed in the water
were used whenever possible.
Correlation: Null hypothesis: the population correlation coefficient (ρ) is equal to zero
Alternative hypothesis: the population correlation coefficient (ρ) is not equal to zero
P values < 0,05 were considered significant
Regression: Null hypothesis: the regression coeficent (β) is equal to zero
Alternative hypothesis: β is not equal to zero
P values < 0,05 were considered significant
Differences between measurements done inside and outside the water were assessed using
the paired t-test.
Null hypotheses: there is no significant difference between measurements performed inside
or outside the water.
Alternative hypothesis: there is a significant difference between measurements performed
inside or outside the water.
P values < 0.05 were considered significant.
Statistical differences between animals with confirmed pulmonary disease and healthy
animals were assessed using the independent 2 sample t-test.
Both the measurements done inside and outside the water were used.
Null hypothesis: the two population means are equal
Alternative hypothesis: the two population means are not equal
P values < 0.05 were considered significant.
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Results
Harbour porpoises
Overview results
In the appendix an overview of the results is shown in tables 5 and 6
Flow/Volume curves
Shown in graphs 4-6 are the flow/volume curves of one healthy animal (Siepy) measured
outside the water, one healthy animal (Sietske) measured inside the water and one animal
(Renske) with a confirmed respiratory disorder.
The remaining flow/volume curves can be found in the attachments.
Graph 4: Flow/Volume curve, Siepy, 29-05-13, measured outside the water, healthy animal
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Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
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Graph 5: Flow/volume curve, Sietske, 02-05-13, measured inside the water, healthy animal
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Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
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Graph 6: Flow/Volume curve, Renske, 02-05-13, measured outside the water, confirmed pulmonary
disorder
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Statistics and graphs
Correlation
In table 7 the results of the linear correlations are shown.
VC
PEF
PIF
Te/Ti ratio
Mean Te
Mean Ti
Mean Vex
Mean Vins
Length in cm
Weight in kg
Age in months
ρ: 0.76
P: 0.01 *
ρ -0.64
P: 0.03 *
ρ: 0,66
P: 0.02 *
ρ: -0.61
P: 0.06
ρ: 0.35
P: 0.26
ρ: 0.50
P: 0.10
ρ: 0.66
P: 0.02 *
ρ: 0.76
P: 0.00 *
ρ: 0.79
P: 0.00 *
ρ: -0.58
P: 0.05
ρ: 0,61
P: 0,04*
ρ: -0.56
P: 0.09
ρ: 0.56
P: 0.06
ρ: 0.65
P: 0.02 *
ρ 0.68
P: 0.02 *
ρ 0.78
P: 0.00*
ρ: 0.64
P: 0.02 *
ρ: -0.56
P: 0.06
ρ: 0.49
P: 0.10
ρ: -0.38
P: 0.28
ρ: 0.17
P: 0.60
ρ: 0.28
P: 0.39
ρ: 0.52
P: 0.08
ρ 0.66
P: 0.02*
Table 7: results of the correlation statistics
* These values are considered statistically significant
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Regression
The data that were proved to correlate were then tested for linear regression.
Graphs 7 and 8 show the VC against the body size of the animals. A positive correlation is
present in both graphs.
Graph 7: VC (l) versus Length (cm)
Intersept (α): -4.17 Standard error of the mean (SEM): 1.68
β: 0.05
SEM: 0.01
P: 0.01 *
P: 0.03 *
Graph 8: VC (l) versus Weight (kg)
α: -1.50 SEM: 0.87 P: 0.11
β: 0.09
SEM: 0.02
P: 0.00 *
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Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
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Graph 9 shows the VC against the age of the animals, there is a positive correlation.
Graph 9: VC (l) versus Age (months)
α:1,11
SEM: 0.39
P: 0.02 *
β:0.02
SEM: 0.01
P: 0.02 *
Graphs 10 – 12 show the maximum flows against the body size of the animals. A negative and two
positive correlations are present.
Graph 10: PEF (l/s) versus Length (cm)
α: 6.48
SEM: 4.93
P: 0.22
β: -0.10
SEM: 0.04
P: 0.03 *
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Graph 11: PIF (l/s) versus Length (cm)
α: -6.45
SEM: 5.37
P: 0.26
β: 0.12
SEM: 0.04
P: 0.02 *
Graph 12: PIF (l/s) versus Weight (kg)
α: 1.05
SEM: 3.11
P: 0.74
β: 0.20
SEM: 0.08
P: 0.04 *
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Graph 13 shows the average Ti against the weight of the animals, a positive correlation is present.
Graph 14 – 18 show the mean volumes of the inspirations and expirations of the animals against
body size and age of the animals. In all these graphs positive correlations are present.
Graph 13: Mean Ti (s) versus Weight (kg)
α: 0.17 SEM: 0.08
P: 0.07
β: 0.01 SEM: 0.00
P: 0.02 *
Graph 14: Mean Vex (l) versus Length (cm)
α: -2.96
SEM: 1.61
P: 0.10
β: 0.04
SEM: 0.01
P: 0.02 *
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Graph 15: Mean Vex (l) versus Weight (kg)
α: -0.95
SEM: 0.86
P: 0.30
β: 0.07
SEM: 0.02
P: 0.02 *
Graph 16: Mean Vins (l) versus Length (cm)
α: -3.44
SEM: 1.37
P: 0.03 *
β: 0.04
SEM: 0.01
P: 0.00 *
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Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
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Graph 17: Mean Vins (L) versus Weight (kg)
α: -1.21
SEM: 0.72
P: 0.12
β: 0.07
SEM: 0.02
P: 0.00 *
Graph 18: Mean Vins (l) versus Age (Months)
α: 0.88
SEM: 0.31
P: 0.02 *
β: 0.02
SEM: 0.01
P: 0.02 *
* These values are considered statistically significant
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Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
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Inside versus outside the water
The paired t-test was used to determine if there were differences in the measurements
performed with the animals in the water versus those done with the animals outside of the
water.
In table 8 the results of the paired t test are shown. df is degrees of freedom.
VC
PEF
PIF
Mean Te/Ti ratio
Mean Te
Mean Ti
Mean Vex
Mean Vins
SD Te
SD Ti
SD Vex
SD Vins
SD Te/Ti ratio
df
t
P
95% confidence interval
8
8
8
4
8
8
8
8
8
8
8
8
4
0,67
0,35
0,72
-1,22
-1,25
0,33
0,48
0,30
-2,50
1,68
0,08
0,20
0,39
0,52
0,74
0,49
0,29
0,25
0,75
0,64
0,77
0,04*
0,13
0,94
0,85
0,72
-0,30 – 0,54
-1,49 – 2,02
-1,01 – 1,92
-0,82 – 0,32
-0,16 – 0,05
-0,09 – 0,12
-0,21 – 0,33
-0,28 – 0,36
-0,06 - -0,002
-0,01 – 0,08
-0,09 – 0,10
-0,12 – 0,14
-0,12 – 0,16
Table 8: results of the paired t-test of the porpoises
*These values are considered significant
In graph 19 the standard deviation of the expiration time of the animals is shown inside and
outside the water.
Graph 19: SD Te versus Location of Measurement (1:Inside, 2:Outside)
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Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
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Animals with confirmed pulmonary disorder versus healthy animals
The independent 2 sample t-test was used to determine if there were differences in the
means of the data obtained by measuring healthy (0-2) animals versus animals with a
confirmed pulmonary disorder (3)
In table 9 the results of the independent 2 sample t-test are shown.
VC
PEF
PIF
Mean Te/Ti ratio
Mean Te
Mean Ti
Mean Vex
Mean Vins
SD Te
SD Ti
SD Vex
SD Vins
SD Te/Ti ratio
df
t
P
95% confidence interval
21
21
21
16
21
21
21
21
20
21
20
21
14
-2,07
7,21
-2,74
4,13
2,48
-2,13
-0,85
-1,72
0,76
-2,16
-1,91
-1,73
0,01
0,05
0,00*
0,01*
0,00*
0,02*
0,045*
0,41
0,10
0,46
0,04*
0,07
0,10
1,00
-2,41 - 0,01
8,82 - 15,97
-9,10 - -1,24
0,41 - 1,28
0,03 - 0,36
-0,28 - -0,003
-1,46 - 0,62
-1,84 - 0,18
-0,04 - 0,08
-0,17 - -0,003
-0,43 - 0,02
-0,45 - 0,04
-0,23 - 0,23
Table 9: results of the 2 sample t-test of the porpoises
*These values are considered significant
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Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
phocoena
In graphs 20 - 25 the differences in the data from animals with a confirmed pulmonary
disorder and the data from healthy animals is shown.
Graph 20: PEF (l/s) versus pulmonary disorder (0: healthy 1: confirmed pulmonary disease)
Graph 21: PIF (L/S) versus pulmonary disorder (0: healthy 1: confirmed pulmonary disease)
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Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
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Graph 22: Mean Te/Ti Ratio versus pulmonary disorder (0: healthy 1: confirmed pulmonary disease)
Graph 23: Mean Te (S) versus pulmonary disorder (0: healthy 1: confirmed pulmonary disease)
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Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
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Graph 24: Mean Ti (S) versus pulmonary disorder (0: healthy 1: confirmed pulmonary disease)
Graph 25: SD Ti versus pulmonary disorder (0: healthy 1: confirmed pulmonary disease)
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Bottlenose dolphins
Overview results
In the appendix an overview of the results is shown in tables 10 and 11
Flow/Volume curves
Shown in graph 26-28 are the flow/volume curves of one healthy animal (Nemo) measured
outside the water, one healthy animal (Tlisala) measured inside the water and one animal
(Tsalka) with a confirmed respiratory disorder.
The remaining flow/volume curves can be found in the attachments.
Graph 26: Flow/Volume curve, Nemo, 30-05-13, measured outside the water, healthy animal
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Graph 27: Flow/Volume curve, Tlisala, 23-05-13, measured inside the water, healthy animal
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Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
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Graph 28: Flow/Volume curve Tsalka, 23-05-13, measured inside the water, confirmed pulmonary
disorder
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Statistics and graphs
Correlation
In table 12 the results of the linear correlations are shown.
VC
PEF
PIF
Te/Ti ratio
Mean Te
Mean Ti
Mean Vex
Mean Vins
Length in cm
Weight in kg
Age in months
ρ: 0.44
P: 0.39
ρ: -0.49
P: 0.32
ρ: 0,39
P: 0,44
ρ: 0.04
P: 0.95
ρ: 0.04
P: 0.98
ρ: 0.14
P: 0.80
ρ: 0.18
P: 0.73
ρ: 0.22
P: 0.68
ρ: 0.23
P: 0.66
ρ: -0.21
P: 0.70
ρ: 0,02
P: 0,97
ρ: -0.32
P: 0.60
ρ: -0.02
P: 0.97
ρ: 0.19
P: 0.72
ρ: 0.12
P: 0.83
ρ: 0.02
P: 0.97
ρ: -0.10
P: 0.86
ρ: -0.04
P: 0.94
ρ: -0,12
P: 0,82
ρ: -0.66
P: 0.23
ρ: -0.39
P: 0.45
ρ: -0.14
P: 0.79
ρ: -0.02
P: 0.97
ρ: -0.21
P: 0.60
Table 12: results of the correlation statistics
* These values are considered statistically significant
No data were statistically significant, so no regression line statistics were done.
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Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
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Inside versus outside the water
The paired t-test was used to determine if there were differences in the measurements
performed with the animals in the water versus those done with the animals outside of the
water.
Table 13 the results of the paired t test are shown.
VC
PEF
PIF
Mean Te/Ti ratio
Mean Te
Mean Ti
Mean Vex
Mean Vins
SD Te
SD Ti
SD Vex
SD Vins
SD Te/Ti ratio
df
t
P
95% Confidence Interval
4
4
4
3
4
4
4
4
4
4
4
4
2
0,29
-2,98
2,78
-1,28
-3,53
-1,0
0,74
1,08
-8,40
-0,58
1,42
0,19
-1,09
0,79
0,04*
0,05*
0,34
0,02*
0,37
0,50
0,34
0,00*
0,59
0,23
0,86
0,39
-1,89 – 2,33
-23,29 – 0,82
0,01 – 6,28*
-0,29 – 0,14
-0,33 - -0,39
-0,27 – 0,13
-0,75 – 1,3
-0,76 – 1,72
-0,07 - -0,04
-0,2 – 0,13
-0,22 – 0,69
-0,24 – 0,28
-0,35 – 0,21
Table 13: results of the paired t-test of the porpoises
*These values are considered significant
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Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
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In graph 29 and 30 the maximum flows of the animals inside and outside the water are
shown.
Graph 29: PEF (L/S) versus Location Measurement (1: Inside, 2: Outside)
Graph 30: PIF (L/S) versus Location Measurement (1: Inside, 2: Outside)
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Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
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In graphs 31 and 32 the mean and standard deviation of the expiration time from the
animals inside and outside the water are shown.
Graph 31: Mean Te (s) versus Location Measurement (1: Inside, 2: Outside)
Graph 32: SD Te (s) versus Location Measurement (1: Inside, 2: Outside)
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Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
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Animals with confirmed pulmonary disorder versus healthy animals
The independent 2 sample t-test was used to determine if there were differences in the
means of the data obtained by measuring healthy (0-2) animals versus animals with a
confirmed pulmonary disorder (3)
In table 14 the results of the independent 2 sample t-test are shown.
VC
PEF
PIF
Mean Te/Ti ratio
Mean Te
Mean Ti
Mean Vex
Mean Vins
SD Te
SD Ti
SD Vex
SD Vins
SD Te/Ti ratio
df
t
P
95% confidence interval
11
11
11
9
11
11
11
11
10
10
10
10
8
3,67
-2,60
2,35
-4,96
-0,58
1,02
3,26
2,60
-0,39
0,74
0,45
2,36
0,08
0,00*
0,03*
0,04*
0,00*
0,57
0,33
0,01*
0,03*
0,71
0,48
0,67
0,04*
0,08
1,62 - 6,49
-24,93 - -2,08
0,50 - 15,05
-0,52 - -0,19
-024 - 0,13
-0,11 - 0,29
0,74 - 3,83
0,47 - 5,66
-0,08 - 0,05
-0,08 - 0,16
-0,56 - 0,83
0,03 - 0,94
-0,30 – 0,02
Table 14: results of the 2 sample t-test of the porpoises
*These values are considered significant
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Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
phocoena
In graphs 33 – 39 the differences in the data from animals with a confirmed pulmonary
disorder and the data from healthy animals is shown.
Graph 33: VC (liters) versus pulmonary disorder (0: healthy 1: confirmed pulmonary disease)
Graph 34: PEF (L/S) versus pulmonary disorder (0: healthy 1: confirmed pulmonary disease)
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Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
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Graph 35: PIF (L/S) versus pulmonary disorder (0: healthy 1: confirmed pulmonary disease)
Graph 36: Mean Te/Ti ratio versus pulmonary disorder (0: healthy 1: confirmed pulmonary disease)
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Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
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Graph 37: Mean Vex (L) versus pulmonary disorder (0: healthy 1: confirmed pulmonary disease)
Graph 38: Mean Vins (L) versus pulmonary disorder (0: healthy 1: confirmed pulmonary disease)
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Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
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Graph 39: SD Vins versus pulmonary disorder (0: healthy 1: confirmed pulmonary disease)
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Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
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Discussion
Having a respiratory system is an unique feature of cetaceans in comparison with other
creatures living in the sea. It is obvious that the respiratory system has undergone major
adaptations in order to function in sea mammals. Long apneustic periods, deep diving with
barometric consequences and very limited time to ventilate are main features necessitating
adaptations. Research has teached us a lot about the respiratory physiology of sea
mammals. 3, 5, 6
This has increased our understanding of the physiologic and morphologic adaptations to a
great extend. However, until now, spirometry in cetaceans has never been systematically
explored. Spirometric data must be significantly altered in comparison to terrestrial animals,
as large volumes must be inhaled and exhaled in a very short period of time. Morphologic
changes in the conducting airways in cetaceans are obvious: the trachi-bronchial system has
a large diameter and has increased rigidity due to circular cartilage, which extends until the
gasexchange zone. 5 Measuring spirometric data will greatly increase our knowledge and
understanding of the ventilatory system of cetaceans. Furthermore as infections of the
respiratory system are common in these species 11, 13 it should be very helpful to use
spirometric measurements as a non-invasive diagnostic tool in screening the respiratory
system for disorders. There are no standard values of spirometry, neither is there a
standardized method of performing these measurements.
Spirometry was performed on harbour porpoises (Phocoena phocoena) as well as bottlenose
dolphins (Tursiops truncates) both while inside the water and out of the water. Because of
obvious logistic limitations this study could only be done in a small number of animals.
Because measurements of the static breathing volumes could not be performed, it cannot
be certain whether the measured VC (vital capacity) is equal tot the TLC (total lung capacity)
or if it is a TV (tidal volume). Earlier research showed the TV of small cetaceans is
approximately 80% of the TLC and therefore can be considered equal to the VC.2
There was some concern about a dead space effect because of the fact that the animals
inhale some of their exhaled air through the hose between the mask and the
pneumotachograph. The mask was disengaged from the animal after two or three breathing
cycles. No differences were measured between consecutive breathing cycles. It was
therefore assumed there was no significant dead space effect.
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Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
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Harbour porpoises
In both species of cetaceans there was an expiratory plateau fase in the flow volume curves
contrary of the situation in humans, where the curves show a downward slope in this
position. 17 This can easily be explained by the absence of dynamic compression at lower
expiratory volumes. The compression is prevented by the existence of cartilage up until the
air exchange zones, which increases rigidity.
In harbour porpoises the PEF is lower than the PIF, whereas in dolphins it is the other way
around. Further more the Te/Ti equilibrium is more than 1.0 in all porpoises and less than
1.0 in all dolphins. An explanation could be found in different morphology of the upper
airways. Apparently the dimensions and the shape of the flow volume curves is dependent
on the species.
The duration of an average breathing cycle is 0,8 seconds and is fairly constant. This means
that the animals are only vulnerable to predator attack for a short period.
The intra individual variation in volumes and times was low, which means that there might
be an opportunity to use this kind of parameters as a personal standard value. This could be
useful when screening for respiratory disorders.
Correlations were found between body size parameters and spirometric data. This is easily
explained as a larger animal breathes a larger volume of air using a higher flow.
There were no marked differences in any measured parameter between animals inside the
water or outside.
The pulmonary function of one sick animal with interstitial lung disease was measured,
which was seen in CT scanning and confirmed by necropsy. In this animal the PIF was lower
than the PEF, and the Te/Ti equilibrium was below 1.0.
The mean Te was lower and the mean Ti was higher than that of the healthy animals.
The duration of the breathing cycle and the VC remained unchanged. These differences are
probably due to altered mechanical properties of the lung tissue.
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Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
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Bottlenose dolphins
The average duration of an exhalation is 0,4 seconds and of an inhalation 0,5 seconds. A
breathing cycle takes therefore about 0,9 seconds. This is longer than was assumed in earlier
literature. 2, 7 The quantity of data is not sufficient to say anything about the influence of
gender. The measurements were very well tolerated. A complicating factor was the fact that
some animals were trained in the past to exhale in a cup for obtaining microbiological
material. They were used to an artificial and extremely powerful style of exhaling. These
measurements were disregarded, though some effect may still be seen from the remaining
measurements. Similar the porpoises, the intra individual variation was low.
In contrast to the results of porpoises, a difference between values measured inside the
water or outside was found. Outside the water the maximum flows were lower and the
mean Te was higher, as wel as the SD of the Te.
The most likely explanation is the difference in weight between porpoises and bottlenose
dolphins. Lying on a rigid structure outside the water the weight of the animal will cause
compression of the dependent lung zones which influences lung function measurements.
This phenomenon should be considered when evaluating lung function measurements in
cetaceans. This is especially true in heavier animals.
Measurements were done in two animals having a pulmonary disorder. One of them had a
tracheobronchitis confirmed with bronchoscopy. The other one was coughing and had an
inflammatory response, and was diagnosed of having pneumonia. This was confirmed by
ultrasound.
The VC, mean Vex and Mean Vins of the sick animals were decreased, as well as the PEF and
the PIF. The length of the total breathing cycle was not changed, but the Te/Ti equilibrium
was consistently increased. The SD of Vins was lower in animals with a confirmed lung
disease, suggesting a flow limitation.
As all animals were about the same size, no correlations were found between size
parameters and spirometric data.
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Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
phocoena
In conclusion, lung function measurements by means of spirometry in cetaceans is a feasible
method to obtain information about lung mechanics. The results of the measurements can
be understood by studying the morphological and physiological features of cetaceans.
The measured volumes are not extremely high in comparison to terrestrial mammals in
correlation to body size. 5 This means that other adaptation methods are vastly more
important in making prolonged apneustic periods possible. There were differences between
the two examined species in the Te/Ti, as well as PEF/PIF equilibrium. This was also seen in
the shape of the flow volume graphs. Results of pulmonary function measurements cannot
be extrapolated from one species to another. In both species animals with confirmed
respiratory disorders were measured. In all of these animals differences were observed in
comparison to healthy animals.
Pulmonary function testing might be helpful in screening and managing respiratory disease
in cetaceans. Both species were measured in the water and on land, in the dolphins there
were marked differences in the maximal flows, which can be explained by compression of
depending lung zones.
It must be emphasized that all results must be evaluated with great caution because of the
very limited number of subjects. Studying larger number of animals of both species and of
both sexes will be helpful to confirm the results of this study. Prospective studies of animals
with respiratory disorders must be performed in order to confirm the usability of spirometry
as a diagnostic tool. The combination of spirometry with other lung function tests may
possibly enhance the clinical usefulness. For example airway resistance measurements and
capnography could be added to spirometric data. Abnormality in lung function will never be
diagnostic without further examining the animal with microbiological investigations or
imaging techniques.
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Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
phocoena
Acknowledgements
First of al I wish to thank the staff of the Dolfinarium for the chance to perform this study, in
particular the trainers of the bottlenose dolphins and the harbour porpoises. They were very
enthusiastic and very helpful.
I wish to thank F.H.C. de Jongh for his help in understanding and processing the data.
I would like to thank the veterinarians of the Dolphinarium, C.E. van Elk and P. Bunskoek, for
their help during the study and for providing the data of the sick animals.
I wish to thank M. G. van Emst, for his support during the study.
I wish to thank N. Boeve-Epping for assistance with the lung function machine.
I wish to thank my family for their moral support.
I wish to thank Kevin van de Bergh and his team for designing and developing the mask.
And last but not least I wish to thank the animals, for their cooperation and (sometimes a bit
too much) enthusiasm.
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Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
phocoena
References
1. Reynolds, J. E., Wells, R. S. & Eide, S. D. in The bottlenose dolphin, biology and conservation 304
(University Press of Florida, 2000).
2. Perrin, W. F., Wursig, B. & Thewissen, J. G. M. in Encyclopedia of marine mammals 1414 (Acedemic
Press, 2002).
3. Ponganis, P. J. Diving mammals. Compr. Physiol. 1, 447-465 (2011).
4. Hoelzel, A. R. in Marine mammal biology an evolutionary approach 431 (Blackwell Science, 2002).
5. Kooyman, G. L. Respiratory adaptations in marine mammals. Integrative and Comparative Biology
13, 457-468 (1973).
6. Snyder, G. K. Respiratory adaptations in diving mammals. Respir. Physiol. 54, 269-294 (1983).
7. Ridgway, S. H. in Mammals of the sea, biology and medicine 812 (Thomas Books, 1972).
8. Lawrence, B. & Schevill, W. E. The functional anatomy of the delphinid nose . Bulletin of the
Museum of Comparative Zoology 14, 103-151 (1956).
9. Reynolds, J. E. & Rommel, S. A. in Biology of marine mammals 578 (Smithsonian Institution, 1999).
10. Sweeney, J. C. & Ridgway, S. H. Common diseases of small cetaceans. J. Am. Vet. Med. Assoc. 167,
533-540 (1975).
11. Dierauf, L. A. & Gulland, F. M. D. in CRC Handbook of Marine Mammal Medicine 1063 (CRC Press,
2001).
12. Eo, K. -. & Kwon, O. -. Two cases of bacterial pneumonia in bottle-nosed dolphins (Tursiops gillii)
at the Seoul Zoo, Korea. Pakistan Veterinary Journal 31, 260-262 (2011).
13. Jeraj, K. P. &Sweeney, J. C. Blowhole Cytology to Diagnose Early Respiratory Tract Disease in
Bottlenose Dolphins (IAAAM 27th Annual Conference Proceedings, Chattanooga, Tennessee, 1996).
14. Gans, S. J. M., van Kregten, E., Marik, A. M., Boeve-Epping, N. & van Elk, C. E. Protected brush
sampling of the respiratory tract of Tursiops truncatus to determine the microbial contamination in
healthy animals (IAAAM 43rd Annual Conference Proceedings, Atlanta, Georgia, 2012).
15. Tsang, K. W. et al. Bronchoscopy of cetaceans. J. Wildl. Dis. 38, 224-227 (2002).
16. Gans, S. J. M., van Elk, C. E., Epping, N., de Jongh, F. H. C. & Hoogsteden, H. C. Lung function
measurements in dolphins; a preliminary report (IAAAM 37th Annual Conference Proceedings,
Nassau, Bahamas, 2006).
17. Ranu, H., Wilde, M. & Madden, B. Pulmonary Function Tests. Ulster Med J 80 (2), 84-90 (2011).
Research project University of Utrecht: S.Gans
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Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
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Attachments
Extra tables
Name
Sex
Date
Age in Length
months in cm
Weight
in kg
Place
Health
Amber
Female 15-05-13
108
154
45,85 Inside
0
Amber
Female 29-05-13
108
154
44,3 Outside
0
Berend
Male
18-03-13
22
118
29,35 Outside
0
Ellen
Female 06-03-13
93
149
50,65 Outside
0
Ellen
Female 19-03-13
93
149
50,65 Inside
0
Ester
Female 06-03-13
11
116
37,4 Outside
0
Ester
Female 01-05-13
12
116
36,05 Inside
1
Gerhard Male
02-04-13
23
116
34 Outside
0
Gerhard Male
02-04-13
23
116
34 Inside
0
Joelle
Female 06-03-13
10
127
36,85 Outside
0
Joelle
Female 01-05-13
12
127
36,95 Inside
1
Jose
Female 15-05-13
36
136
40,15 Inside
0
Jose
Female 15-05-13
36
136
40,15 Outside
0
Siepy
Female 19-03-13
70
147
45,6 Inside
0
Siepy
Female 29-05-13
72
147
42,35 Outside
0
Desiree Female 13-03-13
58
149
50,8 Inside
0
Renske
Female 28-03-13
69
159
76,4 Inside
3
Renske
Female 02-05-13
71
159
67,5 Outside
3
Sietske
Female 28-03-13
21
119
28,3 Inside
2
Sietske
Female 02-05-13
23
119
30,3 Inside
0
Thom
Male
02-05-13
12
105
22,7 Inside
1
Thom
Male
02-05-13
12
105
22,7 Outside
1
Tonia
Female 08-04-13
22
122
31,9 Inside
1
Table 5: overview of the results of the porpoises, data arranged alphabetically
Research project University of Utrecht: S.Gans
VC
in l
PEF in PIF in Mean
l/s
l/s
Te/Ti
3 -9,74 10,78 1,19
1,9
-7,6 11,08 0,8 -4,14 4,58 1,7 -8,52 9,92 2,5 -6,79 8,28 1,18
1,2 -2,78
4,5 1,8
1,2 -4,24 5,32 1,2
2 -5,01
6,4 2 -5,05 7,19 1,42
1,4 -4,12 7,32 1,7
1,2 -3,84 6,52 1,2
2,1 -7,84 10,52 1,07
2,3 -4,84 5,52 1,07
1,7 -5,04 8,44 1,01
1,4 -7,19 9,17 1,93
4,4 -11,36 14,32 1,24
3 -16,32 13,56 0,61
3 -20,64 12,72 0,4
1,3
-6,8 7,24 1,4
1,5
-8,8 10,04 1,13
0,8
-3,6 5,24 1,52
1,5 -4,13 5,11 1,55
1,8 -6,35
9,9 -
48
Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
phocoena
Name
Sex
Date
SD
Te/Ti
Mean SD
Te
Te
Mean SD
Ti
Ti
Mean SD
Vex
Vex
Amber
Female 15-05-13
0,4 0,12 0,34 0,05 2,19 0,52
Amber
Female 29-05-13
0,39 0,09 0,28 0,03 1,57 0,25
Berend
Male
18-03-13
0,32 0,01 0,44 0,19 0,57 0,15
Ellen
Female 06-03-13
0,34 0,08
0,3 0,07 1,15 0,19
Ellen
Female 19-03-13 0,02 0,49 0,1 0,45 0,08 1,83 0,05
Ester
Female 06-03-13 0,14 0,69 0,13 0,42 0,02
0,9 0,25
Ester
Female 01-05-13 0,25 0,53 0,1 0,44 0,08 0,83 0,15
Gerhard Male
02-04-13
0,59 0,16 0,41 0,07 1,61 0,14
Gerhard Male
02-04-13 0,34 0,52 0,11 0,38 0,04 1,43 0,16
Joelle
Female 06-03-13 0,11 0,54 0,16 0,31 0,04 1,08 0,16
Joelle
Female 01-05-13 0,28 0,48 0,1 0,42 0,11 0,88 0,15
Jose
Female 15-05-13 0,09 0,39 0,04 0,37 0,03 1,81 0,25
Jose
Female 15-05-13 0,18 0,68 0,11
0,6 0,03 1,86 0,34
Siepy
Female 19-03-13 0,32 0,54 0,1 0,48 0,22 1,19 0,15
Siepy
Female 29-05-13 0,34 0,49 0,14 0,28 0,04 1,08 0,15
Desiree Female 13-03-13 0,18 0,56 0,12 0,46 0,08 3,67 0,64
Renske
Female 28-03-13 0,21
0,3 0,07 0,48 0,17 1,72 0,44
Renske
Female 02-05-13
0,26 0,08 0,55 0,14 1,93 0,38
Sietske
Female 28-03-13 0,36 0,36 0,08 0,26 0,02 0,99 0,21
Sietske
Female 02-05-13 0,12 0,33 0,03 0,29 0,03 1,41 0,09
Thom
Male
02-05-13 0,18 0,34 0,05 0,21 0,02 0,65 0,08
Thom
Male
02-05-13 0,25 0,52 0,11 0,29 0,04 0,84 0,06
- 0,52
Tonia
Female 08-04-13
0,37 0,14 1,91
Table 6: overview of the results of the porpoises, data arranged alphabetically
Research project University of Utrecht: S.Gans
Mean SD
Vins
Vins
2,48
1,88
0,66
1,2
2
1,2
0,91
1,47
1,63
1,05
0,86
1,85
2
1,36
1,24
3,5
2,13
2,45
1,04
1,35
0,61
0,96
1,35
0,31
0,28
0,13
0,3
0,35
0,19
0,16
0,41
0,17
0,18
0,21
0,27
0,1
0,22
0,09
0,8
0,56
0,33
0,16
0,11
0,04
0,31
0,27
49
Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
phocoena
Name
Sex
Date
Age in Length in Weight
months cm
in kg
Place
Health
VC
in l
PEF in PIF in Mean
l/s
l/s
Te/Ti
Apollo Male
07-05-13
292
285
225 Inside
0 12,8 -48,88 33,28
Apollo Male
30-05-13
292
285
225 Outside
0
10 -22,04 29,64
Juno
Male
07-05-13
352
247
208 Inside
0 7,9 -34,32 24,44
Juno
Male
30-05-05
352
247
208 Outside
0 7,3 -27,16 21,89
Nemo Male
23-05-13
319
262
223 Inside
0 8,1 -26,76 16,64
Nemo Male
30-05-05
319
262
223 Outside
0 9,8 -23,76 17,23
Tlisala Male
23-05-13
148
275
217 Inside
0 7,1 -30,91 25,25
Tsalka Male
09-05-13
184
274
217 Inside
0 11,4 -37,72 22,6
Tsalka Male
23-05-13
184
274
217 Inside
3 4,9 -22,99 17,41
Tsalka Male
30-05-05
184
274
217 Outside
3 4,7 -12,48
11
Tucker Male
07-05-13
388
289
232 Inside
1 8,5 -35,69 22,79
Tucker Male
30-05-05
388
289
232 Outside
1 9,3 -22,94 19,09
Maaike Female 02-11-12
360
270*
266 Inside
3 5,9 -17,08 18,12
Table 10: overview of the results of the bottlenose dolphins, data arranged alphabetically
*No length of this animal was known, this value has been estimated using the lengths of the other
animals.
Name
Sex
Date
SD Mean
Mean SD
Te/ti Te
SD Te Ti
Ti
Mean SD
Vex
Vex
Mean SD
Vins
Vins
Apollo Male
07-05-13 0,22 0,38 0,09 0,68 0,33
8,4
1,9
9
Apollo Male
30-05-13
- 0,62 0,15 0,51 0,14 6,74
1,3 6,96
Juno
Male
07-05-13 0,07 0,27 0,04 0,37 0,07 5,45 0,81 5,91
Juno
Male
30-05-05 0,12 0,41
0,1 0,44 0,17 5,11 0,83 5,08
Nemo Male
23-05-13 0,09 0,41 0,05 0,65 0,09 6,19 0,95
6,6
Nemo Male
30-05-05 0,06 0,43 0,08 0,69 0,12 6,16 0,29
7,1
Tlisala Male
23-05-13 0,11 0,29 0,04 0,35 0,08 4,52 0,46 4,81
Tsalka Male
09-05-13
- 0,43
- 0,66
- 6,83
- 11,42
Tsalka Male
23-05-13 0,06
0,3 0,03 0,32 0,04 3,49 0,53 3,79
Tsalka Male
30-05-05 0,25 0,63 0,08 0,58 0,16 3,85 0,66 3,63
Tucker Male
07-05-13 0,05 0,24 0,02 0,37 0,07 4,82 0,46 5,95
Tucker Male
30-05-05
- 0,42 0,08 0,53 0,18 5,13
0,4 6,08
Maaike Female 02-11-12 0,42 0,38 0,14
0,4 0,1
3,6 0,86 4,06
Table 11: overview of the results of the bottlenose dolphins, data arranged alphabetically
Research project University of Utrecht: S.Gans
1,78
1,57
1,07
1,2
0,9
1,16
0,83
0,62
0,57
1,47
1,25
1,1
50
0,68
0,95
0,76
0,75
0,67
0,66
0,9
1,04
1,09
0,76
1,24
Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
phocoena
Flow volume curves harbour porpoises
Horizontal: volume in liters
Vertical: flow in liters per second
Amber 15-05-13, inside, health: 0
Research project University of Utrecht: S.Gans
51
Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
phocoena
Amber 29-05-13, outside, health:0
Research project University of Utrecht: S.Gans
52
Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
phocoena
Berend, 18-03-13, outside, health:0
Research project University of Utrecht: S.Gans
53
Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
phocoena
Ellen, 06-03-13, outside, health:0
Research project University of Utrecht: S.Gans
54
Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
phocoena
Ellen, 19-03-13, inside, health:0
Research project University of Utrecht: S.Gans
55
Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
phocoena
Ester, 06-03-13, outside, health:0
Research project University of Utrecht: S.Gans
56
Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
phocoena
Ester, 01-05, 13, inside ,health:1
Research project University of Utrecht: S.Gans
57
Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
phocoena
Gerhard, 02-04-13, outside, health:0
Research project University of Utrecht: S.Gans
58
Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
phocoena
Gerhard, 02-04-13, inside, health:0
Research project University of Utrecht: S.Gans
59
Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
phocoena
Joelle, 06-03-13, outside, health:0
Research project University of Utrecht: S.Gans
60
Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
phocoena
Joelle, 01-05-13, inside, health:1
Research project University of Utrecht: S.Gans
61
Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
phocoena
Jose, 15-05-13, inside, health:0
Research project University of Utrecht: S.Gans
62
Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
phocoena
Jose, 15-05-13, outside, health:0
Research project University of Utrecht: S.Gans
63
Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
phocoena
Siepy, 19-03-12, inside, health:0
Research project University of Utrecht: S.Gans
64
Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
phocoena
Siepy, 29-05-13, outside, health:0
Research project University of Utrecht: S.Gans
65
Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
phocoena
Desiree, 13-03-13, inside, health:0
Research project University of Utrecht: S.Gans
66
Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
phocoena
Renske, 28-03-13, inside, health:3
Research project University of Utrecht: S.Gans
67
Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
phocoena
Renske, 02-05-13, outside, health:3
Research project University of Utrecht: S.Gans
68
Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
phocoena
Sietske, 28-03-13, inside, health:2
Research project University of Utrecht: S.Gans
69
Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
phocoena
Sietske, 02-05-13, inside, health:0
Research project University of Utrecht: S.Gans
70
Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
phocoena
Thom, 02-05-13, inside, health:1
Research project University of Utrecht: S.Gans
71
Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
phocoena
Thom, 02-05-13, outside, health:1
Research project University of Utrecht: S.Gans
72
Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
phocoena
Tonia, 08-04-13, inside, health:1
Research project University of Utrecht: S.Gans
73
Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
phocoena
Flow volume curves dolphins
Horizontal: volume in liters
Vertical: flow in liters per second
Apollo, 07-05-13, Inside, health:0
Research project University of Utrecht: S.Gans
74
Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
phocoena
Apollo, 30-05-13, outside, health:0
Research project University of Utrecht: S.Gans
75
Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
phocoena
Juno, 07-05-13, inside, health:0
Research project University of Utrecht: S.Gans
76
Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
phocoena
Juno, 30-05-13, outside, health:0
Research project University of Utrecht: S.Gans
77
Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
phocoena
Nemo, 23-05-13, inside, health:0
Research project University of Utrecht: S.Gans
78
Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
phocoena
Nemo, 30-05-13, outside, health:0
Research project University of Utrecht: S.Gans
79
Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
phocoena
Tlisala, 23-05-13, inside, health:0
Research project University of Utrecht: S.Gans
80
Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
phocoena
Tsalka, 09-05-13, inside, health:0
Research project University of Utrecht: S.Gans
81
Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
phocoena
Tsalka, 23-05-13, inside, health:3
Research project University of Utrecht: S.Gans
82
Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
phocoena
Tsalka, 30-05-13, outside, health:3
Research project University of Utrecht: S.Gans
83
Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
phocoena
Tucker, 07-05-13, inside, health:1
Research project University of Utrecht: S.Gans
84
Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
phocoena
Tucker, 30-05-13, outside, health:1
Research project University of Utrecht: S.Gans
85
Lung function measurements using spirometry in small cetaceans: Tursiops truncatus and Phocoena
phocoena
Maaike, 02-11-12, inside, health:3
Research project University of Utrecht: S.Gans
86