Avian Pathology (February 2006) 35(1), 38
/41

Acute poisoning of silver gulls (Larus novaehollandiae)

following urea fertilizer spillage
Shane R. Raidal1* and Susan M. Jaensch2
1
Division of Veterinary and Biomedical Sciences, Murdoch University, South Street, Murdoch, WA 6150, Australia, and
2
IDEXX Laboratories, PO Box 227, Rydalmere, Sydney, NSW 2116, Australia

Two episodes of accidental urea toxicosis are described in wild silver gulls (Larus novaehollandiae) following
spillage of fertilizer grade urea at a commercial shipping facility near Perth, Western Australia. In both cases,
urea spillage had been seen to contaminate freshwater wash-down pools on the wharves where ships were
being unloaded and gulls were seen to be drinking and washing in the pools nearby the spillages. Affected
birds were found moribund or dead. Necropsy and histopathological findings were non-specific and
consisted of mild to moderate congestion of visceral organs and brain. Analysis of a water sample collected
during Case 1 revealed a very high urea concentration of 4.124 mol/l (pH 5.5), and fluid from the
proventriculus of two birds had urea concentrations of 382 and 308 mmol/l, respectively. Nine birds were
examined during the second episode (Case 2) and, from heparinized heart blood samples collected (n 5), /

the mean plasma urea (288992.0 mmol/l), ammonia (43.9934.2 mmol/l) and uric acid (7.4591.99 mmol/l)
/ / /

concentrations were markedly elevated above the reference ranges for all bird species. Proventricular
contents (n 7) similarly contained high concentrations of urea (3949203 mmol/l) and ammonia (9.3915
/ / /

mmol/l). The probable mechanisms of urea and ammonia toxicity in these birds are discussed.

Introduction

Urea is a commonly used fertilizer in modern agriculture
and it has replaced other major nitrogen fertilizers such
as ammonium nitrate or ammonia because it is non-
explosive and comparatively safe to ship and handle. It is
also less corrosive to equipment and it can be used on
virtually all crops. Accidental acute poisoning by urea
containing fertilizer-contaminated water has been re-
ported in cattle (Horner, 1982; Caldow & Wain, 1991;
Campagnolo et al ., 2002; Villar et al ., 2003), and the
haemotoxic effects of diammonium phosphate and urea
fertilizers has been studied in the walking catfish Clarias
batrachus (Trivedi et al ., 1990). In poultry, accidentally
high concentrations of quaternary ammonium disinfec-
tants can be toxic (Dhillon et al ., 1982) and acute death
in falcons is a recognized risk during the procedure
known as ‘‘Schnather’’, which involves forcing the bird
to ingest ammonium chloride crystals (Samour et al .,
1995). However, there is little documentation of acci-
dental urea fertilizer poisoning of birds. In this paper we
present the clinicopathological findings that were present
in two episodes of acute urea poisoning of wild silver
gulls (Larus novaehollandiae ), associated with spillage of
urea at a shipping terminal.
birds had drunk rainwater contaminated with spilt urea at the shipping
terminal. A water sample collected from shallow puddles on the jetty
and three moribund silver gulls were collected. The birds died within
minutes of being captured and were submitted on ice for necropsy
examination with a post-mortem period of less than 12 h. Tissues
samples of visceral organs and brain were collected, fixed in formalin
and routinely processed for paraffin embedding and histopathological
examination.
Case 2. Dead and moribund silver gulls suspected of having ingested
urea fertilizer-contaminated water were collected by Conservation and
Land Management rangers from the same location as in Case 1 but 4
years later. Urea spillage had been seen to contaminate several
freshwater wash-down pools on the pier and, within hours, approxi-
mately 30 gulls were noticed to be dying after drinking and washing in
the pools near the spillage. Two dead and nine moribund birds were
collected and submitted to Murdoch University Veterinary Hospital for
necropsy examination but all birds had died at the time of accession.
The post-mortem period was estimated to be less than 1 h for all birds
submitted. Plasma samples were obtained from heparinized heart blood
(n/6) centrifuged at 2000 /g for 4 min, and fluid proventricular
contents (n/9) were collected from freshly dead birds and clarified by
centrifugation at 2000 /g for 4 min. These were submitted to determine
urea, ammonia and uric acid concentrations. Samples of visceral organs
and brain were fixed in formalin and routinely processed for paraffin
embedding and histopathological examination.

Materials and Methods

Case 1. About 30 to 40 silver gulls found dead or dying by stevedores at
the Fremantle Port Authority Bulk Cargo Jetty in Kwinana were
investigated by pollution control officers of the Western Australian
Department of Environment Protection. It appeared as though the
Urea, ammonia and uric acid analysis. Urea, ammonia and uric acid
concentrations in water and plasma samples were determined enzyma-
tically using a Cobas Mira (Roche Diagnostics, Rotkreuz, Switzerland)
automatic biochemistry analyser. The ammonia ultraviolet rate reduc-
tion was measured at 340 nm and 378C by the enzymatic conversion of

*To whom correspondence should be addressed. Tel: /61 8 93602418. Fax: /61 8 93104144. E-mail: raidal@murdoch.edu.au
Received 13 July 2005. Provisionally accepted 2 September 2005. Accepted 4 October
ISSN 0307-9457 (print)/ISSN 1465-3338 (online) # 2006 Houghton Trust Ltd
DOI: 10.1080/03079450500465718

ammonia to glutamate in the presence of a-ketoglutarate, NADH and
the glutamate dehydrogenase (GLHD). Urea was similarly measured
enzymatically in a two-step process involving first the enzymatic
hydrolysis of urea to ammonia using urease and then the conversion
of ammonia to glutamate in the presence of a-oxoglutarate, NADH and
GLDH. Uric acid was measured enzymatically by the modified Trinder
peroxide method (Trinder, 1969).
Results
In Case 1, analysis of the environmental water sample
revealed a very high urea concentration of 4.124
mol/l (pH 5.5) and fluid from the proventriculus
of two birds had urea concentrations of 382 and 308
mmol/l, respectively. Histopathological examination of
the brain and visceral organs demonstrated mild to
moderate diffuse congestion and low numbers of uni-
dentified cestodes in the alimentary tract. In one bird the
ureters were distended with unidentified adult trema-
todes associated with a moderate proliferative, lympho-
plasmacytic and heterophilic pyelonephritis.
Two birds in Case 2 had evidence of minor scavenging,
and the proventriculus of four birds was distended with
watery fluid. There was mild to moderate congestion of
visceral organs and the brain. Urea and ammonia
concentrations in heart blood and proventricular con-
tents were beyond the limits of the assay and needed to
be diluted 1:100. From heart blood samples (n/5) the
mean plasma urea (2889/92.0 mmol/l), ammonia (43.99/
34.1 mmol/l) and uric acid concentrations (7.459/1.99
mmol/l) were markedly elevated (Table 1). The stomach
contents (n/7) similarly contained markedly high con-
centrations of urea (3949/203 mmol/l) and ammonia
(9.39/15 mmol/l).
Histological examination demonstrated in three birds
a periportal pattern of mild microvesicular cytoplasmic
vacuolation of hepatocytes and moderate to marked
sinusoidal congestion in the liver. There was moderate to
marked congestion of other visceral organs and the
brain. Acute mild to moderate renal tubular degenera-
tion was present in the kidneys of two birds. Cross-
sections of occasional unidentified cestodes were present
in the small intestine of several birds and two birds had a
moderate lymphoplasmacytic enterocolitis associated
with moderate numbers of schistosome eggs. One of
these birds also had adult schistosomes present in major
pancreatic vessels consistent with Austrobilharzia terre-
galensis infection.
Table 1. Plasma and proventricular fluid urea, ammonia and uric acid concentrations in nine silver gulls (Larus novaehollandiae)
Urea (mmol/l)
Sample (normalB/1 mmol/l)
Bird 1 Stomach
Bird 1 Plasma
Bird 2 Plasma
Bird 3 Stomach
Bird 3 Plasma
Bird 4 Plasma
Bird 5 Stomach
Bird 6 Plasma
Bird 6 Stomach
Bird 7 Stomach
Bird 8 Stomach
Bird 9 Stomach
Urea poisoning in gulls 39
Discussion
Pathological examination of dead and dying birds can
provide important documentation of toxicological con-
tamination of the environment (Smit, 1980). The clinical
signs, massively elevated proventricular and plasma urea
concentrations compared with published reference
ranges for all bird species (Lumeij & Overduin, 1990;
Lumeij, 1994; Fudge, 2000) and histopathological find-
ings present in the gulls in both cases presented above,
were consistent with the observations of the birds having
been inadvertently poisoned with urea fertilizer-con-
taminated water. The urea concentrations seen in these
gulls would have had a serious detrimental effect on
protein function throughout the body (Morgan et al .,
2003). Marine elasmobranchs are the only animals that
are adapted to cope with plasma urea concentrations in
the 300 to 600 mM range (Part et al ., 1998; Withers et
al., 1994; Fines et al ., 2001), and they do so by
accumulating methylamines and other low-molecular-
weight osmolytes that offset the effects of urea on
proteins (Lee et al ., 1991; Withers et al., 1994).
Urea is readily hydrolysed to ammonia with exposure
to water and/or heat (Denmead et al ., 2004) and this
process can be accelerated by bacterial ureases in the
environment and gut (Pathe et al ., 2003). The concen-
trations of ammonia in the proventricular fluid (0.4 to 95
mmol/l) of affected birds was likely to be predominantly
due to the environmental hydrolysis of urea, while the
concentrations in the blood (1.2 to 36 mmol/l) would
reflect a combination of environmental and in-vivo
hydrolysis. Both urea and ammonia are rapidly absorbed
intact from the avian gastrointestinal tract (Karasawa,
1984; Karasawa & Koji, 1994). The rapid uptake of
ammonia expected with this gastrointestinal loading
would exceed the maximal renal excretion rate of
ammonia of 500 mmol/h per kg body weight (Long,
1982), and would result in rapid elevation of plasma
levels. Such high plasma concentrations of ammonia
alone would be enough to cause death in most animal
species.
Ammonia (NH3) is a natural byproduct of amino acid
metabolism and, because it is a base, it rapidly accepts an
H ion to form ammonium (NH4) at physiological pH
(Cameron & Heisler, 1983). Both ammonia and ammo-
nium are toxic and an effective excretion system is
essential to maintain cellular function. Hydrated NH4
has the same ionic radius as hydrated K ion (Knepper
et al ., 1989), and, due to its K -like behaviour, NH4
Ammonia (mmol/l) Uric acid (mmol/l)
(normalB/0.5 mmol/l) (normalB/1 mmol/l)
580 62.1
/
360 1.2 6.67
310 2.6 10.74
120 95.0
/
380 5.2 5.56
230 36.0 7.70
580 59.0
/
160 1.5 6.59
380 13.90
/
350 60.9
/
150 16.9
/
600 0.4
/
40 S. R. Raidal and S. M. Jaensch
can affect the membrane potential of neurons and other
cells (Binstock & Lecar, 1969; Cooper & Plum, 1987) as
well as result in toxic accumulations of glutamine and
disruption of essential cellular functions. Across the
animal kingdom the ammonia concentration of body
fluids is typically low (50 to 400 mmol/l) (Cooper &
Plum, 1987; Wood et al ., 2002; Cameron & Batterton,
1978; Weihrauch et al ., 1999), with avian normal plasma
ammonia ranging from 48 to 869 mmol/l in granivorous
and omnivorous birds (Karasawa & Kibe, 1983; Kar-
asawa & Nakata, 1988; Boren et al ., 1996). Concentra-
tions exceeding 1 mmol/l total ammonia (NH3 and
NH4) are usually toxic to mammalian cells (Hrnjez et
al ., 1999), and toxic effects have been identified in avian
lymphocytes at concentrations as low as 300 mmol/l
(Klucinski & Targowski, 1984).
The markedly elevated plasma uric acid concentra-
tions (7.459/1.99 mmol/l) observed in the birds from
Case 2 were 5 to 10 times the nominal reference range for
passerine birds (Fudge, 2000), most probably due to an
inadequate attempt by the liver to convert and excrete
the subjected ammonia load, with concurrent suppres-
sion of renal uric acid excretion by the ammonia
(Karasawa & Kibe, 1983). In birds excretion of metabo-
lically derived ammonia is achieved by first converting it
to the amino acids glutamine (via glutamate), glycine
and aspartate for incorporation into the purine synthetic
pathway. This pathway is much more energy expensive
than the urea cycle of mammals, with the main
advantage being the excretion of relatively insoluble
nitrogen-rich uric acid that, being about 40,000 times
less soluble than urea, is not reabsorbed from the cloaca
in the adult or from the allantois in the avian embryo.
Birds do not lack any of the urea cycle enzymes, and
arginine, an essential amino acid in birds, can still be
broken down by arginase to form urea. However,
arginase is predominantly mitochondrial and not cyto-
plasmic as in mammals and the highest concentrations
occur in the kidney rather than the liver (Traniello et al .,
1975; Kadowaki et al ., 1976; Kadowaki & Nesheim,
1978). Avian arginase can be upregulated in birds fed
high-protein diets and is inhibited by adenosine tripho-
sphate, so when energy is not limited urea is not
produced (Ruiz-Feria et al ., 2001; Koutsos et al .,
2001). In low ambient temperatures, some nectarivorous
birds may conserve energy by increasing urea excretion
and lowering uric acid excretion; hummingbirds can
even excrete up to 50% of waste nitrogen as ammonia
provided that water intake is not limited (Preest &
Beuchat, 1997; Roxburgh & Pinshow, 2002; McWhorter
et al ., 2003). However, this has not been demonstrated in
non-nectarivorous species (Sabat et al ., 2004).
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