Avian Pathology

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Haematology of wild penguins (spenisciformes) in

the falkland islands

C.M. Hawkey , D.T. Horsley & I.F. Keymer

To cite this article: C.M. Hawkey , D.T. Horsley & I.F. Keymer (1989) Haematology of wild
penguins (spenisciformes) in the falkland islands, Avian Pathology, 18:3, 495-502, DOI:
10.1080/03079458908418621

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Avian Pathology, 18: 495-502, 1989

HAEMATOLOGY OF WILD PENGUINS (SPENISCIFORMES)

IN THE FALKLAND ISLANDS

C.M. HAWKEY1, D.T. HORSLEY2 and I.F. KEYMER3

1
Department of Veterinary Science, Institute of Zoology, Zoological
Society of London, Regents Park, London, NW1 4RY, England
2
Gresham's School, Holt, Norfolk, England
3
Edgefield, Melton Constable, Norfolk, NR24 2AL, England
SUMMARY
Haematological values were determined in 50 Rockhopper (Eudyptes
crestatus), 19 Gentoo (Pygoscelis papua) and 12 Magellanic (Spheniscus
magellanicus) penguins from various sites on the Falkland Islands.
Adult Magellanic penguins had significantly lower haemoglobin (Hb)
levels, packed cell volumes (PCV) and red cell counts (RBC) than
adults of the other two species. Hb, PCV and RBC values were also
lower in juvenile birds than in adults and lower in post-moult than in
pre-moult adults. Comparison of findings in wild Rockhopper and
Gentoo penguins with values obtained from captive birds showed slight
but significant differences in Hb and mean cell haemoglobin concen-
tration, and in the relative numbers of heterophils, lymphocytes,
monocytes and eosinophils present.

INTRODUCTION
The Sphenisciformes is one of the numerous avian orders for which little haemato-
logical information is available. Published studies on wild penguins include descrip-
tions of red cell parameters and differential white cell counts in Gentoo {Pygoscelis
papua), Chinstrap (P. antárctica) and Adelie (P. adeliae) penguins (Block and Murrish,
1974; Guard and Murrish, 1975; Myrcha and Kostelecka-Myrcha, 1980; Murrish,
1982; Zinsmeister and Van Der Heyden, 1987). Reference values for captive
adult Gentoo, Rockhopper {Eudyptes crestatus), King {Aptenodytes patagónica),
Black-footed {Spheniscus demersus) and Humboldt's {S. humboldti) penguins were
reported by Hawkey and Samour (1988). Seasonal and age-related haematological
changes have been investigated in wild pygoscelid penguins (Kostelecka-Myrcha and
Myrcha, 1980) and in captive Black-footed penguins (Stoskopf et al, 1980).
Limited clinical studies, including descriptions of lymphocytosis associated with
malarial infections in captive Black-footed penguins (Stoskopf and Beier, 1979) and

Received 29 July 1988

Accepted 12 December 1988
496 CM. Hawkey et al

of haematological changes in Gentoo penguins with bumblefoot (Hawkey et al,

1985) indicate that haematological testing is of diagnostic value in these birds and
studies on penguin physiology suggest interesting environmental adaptations
involving their blood cells (Block and Murrish, 1974; Guard and Murrish, 1975).
Acquisition of additional haematological data on members of the Sphenisciformes
is therefore of particular interest.
During a recent investigation of penguin mortality in the Falkland Islands (Keymer,
1987), the opportunity arose to obtain and test blood samples from a number of
clinically healthy Rockhopper, Gentoo and Magellanic (Spheniscus magellanicus)
penguins. Here are reported the results of the study and, where possible, the
findings are considered in relation to age, stage of moult and in comparison with
results on captive penguins of the same species.

MATERIALS AND METHODS

The study was carried out on 50 Rockhopper, 19 Gentoo and 12 Magellanic pen-
guins from various sites on the Falkland Islands during the 1986-87 breeding
season. Blood samples were obtained from the brachial vein of birds manually
restrained in a specially designed cylindrical holder, using disposable needles and
plastic syringes (Samour et al, 1983) and placed in commercially available plastic
tubes containing lithium heparin (100 IU/ml of blood). Samples were also obtained
for plasma biochemical tests and vitamin assays, the results of which are reported
elsewhere (Ghebremeskel er al, 1988a, b; Williamsei al, 1988).
Haematological tests on wild birds were carried out using field methods. Haemo-
globin (Hb) and red cell count (RBC) were estimated using a Compur Mini-
Photometer, Model Ml 100 (Centronic Sales Ltd). Because the nuclei of avian red
cells remain in suspension in the Drab kins solution used in this technique, it was
necessary to modify the technique by the addition of 20 /ul of 0.88 ammonia to
each cuvette. The Hb level was then read directly as oxyhaemoglobin. Red blood
cells were counted according to the manufacturer's instructions but, because the
results obtained are influenced by red cell size, recorded counts were corrected by
reference to a curve prepared in advance under laboratory conditions, using blood
samples obtained from healthy black-footed penguins. Comparison of RBC values
from wild penguins obtained in this way was considered to be valid but comparison
with results from captive penguins, obtained by standard laboratory techniques was
not done since validation was not possible.
Packed cell volume (PCV) was measured using a Compur Minihaematocrit centri-
fuge, Model Ml 101 (Centronic Sales Ltd).
Blood films were fixed in absolute methanol and transported to the laboratory for
staining and examination.
Haematological values on captive penguins were obtained from healthy birds in the
collection of the Zoological Society of London, using standard laboratory tech-
niques (Hawkey and Samour, 1988). Mean cell volume (MCV) and mean cell
haemoglobin concentration (MCHC) were calculated from standard formulae.
Slides were stained by the May Grunwald-Giemsa method. For normally distributed
data, significant differences (P<0.05) were determined using the one-way analysis
of variance and the r-test for two means. Values for percentage monocytes,
 Penguin haematology 497

eosinophils and basophils, which are not normally distributed, were compared by
the Kruskal-Wallis one-way analysis of variance.

RESULTS
Red cell values and differential white cell counts for wild adult penguins are shown
in Table 1. Significant differences between species were found for Hb (¿><0.01),
RBC (P = 0.01), MCV (P = 0.01) and MCHC (P= 0.01). These values did not differ
significantly between Gentoos and Rockhoppers but Magellanics had lower Hb
(P = 0.01), RBC (P = 0.01) and MCHC (P0.001) levels and a higher MCV (¿>= 0.02)
value than Gentoos and Rockhoppers. The relative number of heterophils
(P = 0.001) was significantly higher in Rockhoppers than in Gentoos. Blood para-
sites were not found.

Table 1. Haematological values on wild adult Rockhopper, Gentoo and Magellanic

penguins
Variable Rockhopper Gentoo Magellanic
Mean ± SD Mean ± SD Mean ± SD
Hb (g/dl) 16.4 ±2.7 17.2 ±2.6 13.9 ± 1.8
RBCixlO 12 / 1 ) 2.36 ±0.43 2.24 ±0.41 1.99 ±0.42
PCVU/1) 0.45 ± 0.05 0.45 ± 0.06 0.42 ± 0.04
MCV (fl) 195 ±29 206 ± 29 215 ±34
MCHC (g/dl) 36.6 ±4.1 37.8 ±3.5 33.1 ±2.8
Hetero (%) 29 ±8 41 ± 14 32 ± 14
Eos (%) 6.4 ±5.3 4.5 ±3.6 6.4 ±4.2
Basos {%) 0.6 ±1.2 0 0.1 ±0.3
Lymphs 63 ± 10 54 ± 15 60 ± 17
Monos 1.8 ± 1.0 0.9 ± 1.0 1.2 ± 1.1
n 33 - 43 13 -14 9 - 12
Hb = haemoglobin; RBC = red cell count; PCV = packed cell volume; MCV = mean
cell volume; MCHC = mean cell haemoglobin concentration; Hetero = heterophils;
Eos = eosinophils; Basos = basophils; Lymphs = lymphocytes; Monos = monocytes.

Comparison between adult and juvenile birds

Findings from wild juvenile Rockhopper and Gentoo penguins are shown in Table
2. Values for juvenile Magellanic penguins were not available. In both species
examined, the values for Hb, RBC and PCV were significantly higher in adults
than in juveniles but age differences in MCV and MCHC were not significant.
Influence of moulting
Wild adult Rockhopper penguins had significantly lower Hb, RBC, PCV, MCHC
and heterophil values and raised MCV and lymphocyte values at the end of the
moulting period (Table 3). The fall in MCHC was present during the moulting
period. Magellanic penguins showed the same trends but these were statistically
significant only for Hb and MCHC (Table 4). Moulting Gentoo penguins were not
studied.
498 CM. Hawkey et al.

Table 2. Haematological findings in wild juvenile Rockhopper and Gentoo penguins

Rockhopper • Gentoo
Mean±SD P* Mean ± SD P*
Hb (g/dl) 14.0 ±2.1 <0.05 13.1 ±2.1 <0.01
RBCtxlO12/1) 1.95 ±0.52 <0.05 1.62 ±0.24 <0.01
PCVU/1) 0.38 ±0.07 <0.01 0.37 ±0.05 <0.05
MCV (fl) 201 ±28 NS 232 ±25 NS
MCHC (g/dl) 37.1 ±3.8 NS 35.1 ±2.9 NS
n 1 5

* Compared with adult values in Table 1. Abbreviations as for Table 1.

Table 3. Influence of moulting on haematological values of adult Rockhopper

penguins

Pre-moult During moult Post-moult

Mean ± SD Mean ± SD P* Mean ± SD P*

Hb (g/dl) 18.4 ± 1.4 17.8 ± 1.9 NS 12.2 ± 1.6 <0.001

RBCUIO 1 2 / 1 ) 2.53 ±0.19 2.78 ±0.17 NS 1.59 ±0.32 <0.001
PCV (1/1) 0.46 ± 0.03 0.47 ± 0.04 NS 0.37 ±0.06 <0.001
MCV (fl) 184 ± 11 193 ±22 NS 236 ±28 <0.001
MCHC (g/dl) 39.7 ±3.6 35.4 ±3.2 <0.01 33.0 + 2.4 0.001
Hetero (%) 35 ± 9 27 ±9 NS 21 ± 7 <0.01
Eos (%) 7 ±6 8 +5 NS 3 ±3 NS
Basos (%) 1 ±1 1 ±1 NS 1 ±1 NS
Lymphs (%) 55 ± 12 62 ±8 NS 74 ± 7 <0.005
Monos (%) 2 ±2 2 +2 NS 2 ±2 NS
n 9-11 19 7

Compared with pre-moult values. Abbreviations as for Table 1.

Table 4. Influence of moulting on haematological values of wild adult Magellanic

penguins

Pre-moult Post-moult
Mean ± SD Mean ± SD P*

Hb (g/dl) 14.9 ± 1.8 12.6 ±0.5 <0.05

RBC(xlO 1 2 /') 2.14 ±0.49 1.78 ±0.15 NS
PCV (1/1) 0.42 ± 0.04 0.41 ±0.03 NS
MCV(fl) 204 ± 38 231 ±23 NS
MCHC (g/dl) 34.9 ± 1.9 30.9 ±2.0 <0.01
n 10 - 12 7

Compared with pre-moult values. Abbreviations as for Table 1.

 Penguin haematology 499

Comparison of wild and captive birds

Findings on captive adult Rockhopper and Gentoo penguins are shown in Table 5.
Information for captive adult Magellanics was not available. Values for RBC and
MCV were not included in this comparison because the method used for measuring
the RBC may not have been strictly comparable in the two groups. Compared with
captive Rockhoppers, wild Rockhoppers had significantly lower Hb and MCHC
values. Red cell differences between captive and wild Gentoos were not significant.
The relative number of heterophils was lower and the relative number of lympho-
cytes, monocytes and eosinophils higher in wild than in captive birds of both
species (Table 5).

Table 5. Haematological values for captive adult Rockhopper and Gentoo penguins
Rockhopper Gentoo
Mean ± SD P* Mean ± SD P*

Hb (g/1) 17.9 ± 1.1 <0.05 16.0 ± 1.4 NS

PCV (1/1) 0.47 ± 0.02 NS 0.44 ± 0.06 NS
MCHC (g/dl) 39.2 ±0.8 <0.05 36.5 ±3.0 NS
Hetero (%) 69 ± 12 <0.001 69 ± 10 <0.001
Eos (%) 1.1 ± 1.5 0.001 0.5 ± 1.7 0.002
Basos (%) 0.6 ±0.2 NS 0 NS
Lymphs (%) 27 ± 14 <0.001 30 ± 10 <0.001
Monos 0.1 ±0.3 <0.001 0 <0.01
n 12 - 15 10 - 14

Compared with adult values in Table 1. Abbreviations as for Table 1.

DISCUSSION
The limited information available on the haematology of penguins indicates that
the Hb and PCV values in healthy adults are high compared with those of most
other birds and that penguin red cells are amongst the largest so far found in
birds (Block and Murrish, 1974; Guard and Murrish, 1975; Myrcha and Kostelecka-
Myrcha, 1980; Bennett and Hawkey, 1988; Hawkey and Samour, 1988). The adult
Rockhopper and Gentoo penguins investigated in our survey also had relatively
high mean Hb and PCV levels, although significantly lower values were found in the
Magellanics. Ectoparasites are common in Magellanic penguins which, unlike
Rockhoppers and Gentoos, nest in burrows and the low Hb and comparative
hypochromia may have been caused by chronic blood loss due to ectoparasitism
because 50% of 12 Magellanics examined during the survey harboured ectoparasites
compared with only 9% of 67 Rockhoppers and none of the Gentoos. Ticks from
the Magellanics were identified as Ixodes (eratixodes) urie, White, 1852, syn /.
putus, Pickard-ambridge, 1876 and fleas from the Rockhopper as Parapsyllus
longicornis alginus, Jordon, 1942.
The MCV values recorded in all three species confirmed that penguins have rela-
tively large red cells; this observation may have interesting phylogenetic or eco-
logical implications (Bennett and Hawkey, 1988). The finding of lower Hb, RBC
500 CM. Hawkey et al.

and PCV values in juvenile compared with adult Rockhoppers and Gentoos is
consistent with reports of similar age-related differences in captive Black-footed
penguins and wild pygoscelid penguins (Kostelecka-Myrcha and Myrcha, 1980;
Stoskopf et al, 1980) and in other avian species including domestic fowl (Gallus
gallus) and geese (Amer spp.), quail (Cotumix spp.) and rosy (Phoenicopterus
ruber) and Chilean (P. chiliensis) flamingos (Hodges, 1977; Hawkey et al, 1984).
This presumably reflects the failure of erythropoietic activity to match growth
rate during the juvenile period.
Because a field technique for undertaking total white cell counts was not available,
it is not possible to comment on white cells in terms of their absolute numbers. It
was interesting to note that, in wild birds, the lymphocyte was the most numerous
white cell present (Table 5) whereas, in captive birds, heterophils outnumbered
lymphocytes (Table 1). This finding is in contrast to that of Zinsmeister and
Van Der Heyden (1987) who found more heterophils than lymphocytes in wild
Gentoo, Chinstrap and Adelie penguins. As a general rule, the absolute lympho-
cyte count is relatively stable in healthy adult birds while the heterophil count,
which can fluctuate under the influence of stress and microbial challenge, is much
more susceptible to variation at both the individual and the species level (Hawkey
and Samour, 1988). These observations raise the possibility that, in the birds
examined in this survey, exposure to bacteria was more common in the captive
group. However, the alternative possibility that lymphocytosis was present in the
wild birds cannot be ruled out.
The haematological differences found in pre- and post-moult Rockhoppers and
Magellanic penguins were consistent with those expected during a period of starva-
tion and increased protein utilisation for feather production. Relatively low Hb
levels were recorded at the end of the moult. In both species, this gave rise to low
MCHC values and, in the Magellanics, the degree of hypochromia was greater than
generally expected in healthy birds (Hawkey and Bennett, 1988). Parallel measure-
ments of plasma biochemical constituents in the same birds showed significantly
lower post-moult levels of total protein, albumin, iron, urate and potassium
(Ghebremeskel et al, 1988a). The post-moult reduction in mean plasma iron was
greater than 70% in both species, possibly as a result of plasma transferrin deple-
tion by utilisation during the moult-fast (Ghebremeskel et al, 1988a). Reduced
post-moult Hb and RBC values have been reported in mallard drakes (Anas platy-
rhynchos), but in these birds, which also show a 30% weight loss during moulting
although their food intake is increased (Woods, personal communication), the
MCHC and total plasma protein level were not affected (Driver, 1981). Clearly,
the physiological demands imposed by moulting are considerable.
Acknowledgements
We are grateful to the Falkland Islands Government and the World Wildlife Fund
for financing the expedition and to the Falkland Islands Development Corporation
(FIDC) for organisational help. Messrs Shane Wolsey and David Morgan of the
FIDC also helped considerably with the field work and Mr F. Cheong (Agricultural
Research and Development Centre) provided laboratory facilities in the Falklands.
The ticks were kindly identified for us by Dr K.H. Hyatt and the fleas by Annette
Thomas, both of the British Museum (Natural History), London.
 Penguin haematology 501

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logical aspects in mammals and birds. In: Animal Clinical Biochemistry: The Future,
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Antarctic Journal of the United States, 9: 98-99.
Driver, E.A. (1981). Hematological and blood chemical values of mallard, Anas p. platyrhyn-
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Ghebremeskel, K., Williams, C., Keymer, I.F., Horsley, D.T. and Gardner, D.A. (1988a).
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RESUME
Etudes hématologiques des pingouins sauvages
(Sphenisciformes) dans les îles Falkland
Les valeurs hématologiques ont été déterminées chez 50 gorfous sauteurs (Eudyptes
crestatus), 19 manchots papous (Pygoscelis papua) et 12 manchots de Magellan
(Spehniscus magellanicus) de différents sites des îles Falkland. Les pingouins
Magellanic adultes avaient des taux d'hémaglobines (Hb), des volumes globulaires
(PCV) et un nombre d'hématies (RBC) significativement inférieurs à ceux des
adultes des deux autres espèces. Les valeurs Hb, PCV et RBC étaient également
inférieures chez les jeunes comparés aux adultes, et chez ces derniers plus basses
après la mue qu'avant la mue. La comparaison des valeurs obtenues chez les pin-
gouins sauvages Rockhopper et Gentoo avec celles d'oiseaux captifs ont révélé
de légères différences non significatives en ce qui concerne le taux Hb, ainsi que
le nombre relatif d'hétérophiles, de lymphocytes, de monocytes et d'éosinophiles.

ZUSAMMENFASSUNG
Haematologische Untersuchungen bei wilden Pinguinen
(Sphenisciformes) der Falklandinseln
Haematologische Werte wurden bei 50 Felsenpinguinen (Eudyptes crestatus),
19 Eselspinguinen (Pygoscelis papua) und 12 Magellanpinguinen (Spheniscus
magellanicus) an verschiedenen Örtlichkeiten der Falklandinseln bestimmt. Erwach-
sene Magellanpinguine hatten signifikant niedere Haemoglobin (Hb) Werte und ein
niederes gepacktes Zellvolumen (PCV) sowie geringere Zahlen Roter Blutzellen
(RBC) als erwachsene Exemplare der zwei anderen Spezies. Hb, PCV und RBC-
konzentrationen waren auch bei den juvenilen Vögeln geringer als bei erwachsenen
und bei erwachsenen Tieren nach der Mauser geringer als vor dieser. Der Vergleich
der Befunde von wilden Felsenpinguinen und Eselspinguinen mit den Werten
gefangener Vögel ergab geringe aber signifikante Unterschiede bei Hb und der
mittleren Haemoglobinzellkonzentration sowie bei den relativen Heterophilen-,
Lymphozyten-, Monozyten- und Eosinophilenzahlen.

RESUMEN
Estudios hematológicos en los pingüinos salvajes
(Esfenisciformes) de las islas Malvinas
Se determinaron los valores hematológicos en 50 pingüinos roqueros (Eudyptes
crestatus), 19 pingüinos pájaro bobo (Pygoscelis papua) y 12 pingüinos del Maga-
llanes (Spheniscus magellanicus), procedentes de varias zonas de las islas Malvinas.
Los individuos adultos del pingüino del Magallanes tuvieron los niveles de hemo-
globina (Hb), el hematocrito (PCV) y el número de eritrocitos significativamente
más bajos que los de las otras dos especies. Dichos valores fueron también más
bajos en las aves jóvenes que en las adultas e inferiores en los adultos después de la
muda que antes de la misma. Hubo una diferencia ligera, aunque significativa, en
los niveles de Hb y de concentración media celular de hemoglobina, así como en el
número relativo de heterófilos, linfocitos, monocitos y eosinófilos entre los pingüi-
nos roqueros y pájaro bobo salvajes y las aves cautivas.