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Toxicon 136 (2017) 1e5
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Toxicon

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Case report
Massive attack of honeybee on macaws (Ara ararauna and Ara
chloropterus) in Brazil e A case report

Elisane Lenita Milbradt a, *, Tarcísio Macedo Silva a, Alessandre Hataka a,
Carlos Roberto Teixeira b, Adriano Sakai Okamoto a, Raphael Lucio Andreatti Filho a

a Department of Veterinary Clinical Sciences, School of Veterinary Medicine and Animal Science, S~ao Paulo State University (UNESP), Botucatu, S~ao Paulo,
Brazil
b Department of Veterinary Surgery and Anesthesiology, School of Veterinary Medicine and Animal Science, S~ao Paulo State University (UNESP), Botucatu,
S~ao Paulo, Brazil
a r t i c l e i n f o

Article history:
Received 20 April 2017
Received in revised form
12 June 2017
Accepted 13 June 2017
Available online 16 June 2017

Keywords:
Honeybee
Envenomation
Lesions
Histopathology
Avian pathology
Macaw
* Corresponding author. Universidade Estadual Paul
- UNESP, Faculdade de Medicina Veterin�aria e Zootec
Walter Maurício Correa s/n, UNESP Campus de Botuc
Veterin�aria, Laborat�orio de Ornitopatologia, Botucatu
Brazil.

E-mail address: emilbradt@gmail.com (E.L. Milbra

http://dx.doi.org/10.1016/j.toxicon.2017.06.007
0041-0101/© 2017 Elsevier Ltd. All rights reserved.
a b s t r a c t

Three adult birds of the species Ara chloropterus and five of the species Ara ararauna from a conservation
breeding facility suffered a massive attack by honeybees. The A. chloropterus birds presented swollen
puncture lesions with stingers (mainly in the facial regions without feathers), swelling of the eyelids and
subcutaneous tissue, and respiratory distress, and they were treated with intramuscular injections of 1.67
mg/kg of promethazine and 10 mg/kg of hydrocortisone followed by removal of the stingers. Complete
remission of the clinical signs occurred 48 hours after start of treatment. The five A. ararauna birds died
before they arrived at the veterinary hospital, and the necropsies found stingers in the areas of the face
without feathers and the subcutaneous tissue, which were associated with erythema, bruising, and
swelling. Food content from the crop was found in the oral cavity and the tracheal lumen, and marked
congestion was observed in the heart, liver, spleen, lungs, kidneys, brain, and cerebellum. Among the
histopathological findings, significant swelling of the myocytes in the endocardium and vascular dilation
with erythroid repletion were observed, and there were multifocal areas of centrilobular necrosis
associated with severe congestion and hemorrhaging in the hepatic tissue. Severe acute tubular necrosis
and hydropic-vacuolar degeneration were observed in the kidneys. The clinical signs and pathological
findings suggest envenomation due to a massive bee attack, the first such report for Psittacidae.

© 2017 Elsevier Ltd. All rights reserved.
1. Introduction

Brazil may be considered the cradle of the hybrid Africanized
honeybee since African bees (Apis mellifera scutellata) imported in
the 1950s were cross-bred with European species (Apis mellifera
and Apis mellifera ligustica). The crossbreeding resulted in a bee that
produces more honey than the African bee but displays more
aggressive behavior than the European species, and this has led to
various types of accidents with severe and even fatal results for
both animals and humans (Almeida et al., 2011; Clarke et al., 2002;
Ferreira et al., 2012; Funayama et al., 2012). This hybrid spread
ista “Júlio de Mesquita Filho”
nia - FMVZ, Rua Prof. Doutor
atu, Departamento de Clínica
, S~ao Paulo, CEP: 18618-681,

dt).
quickly throughout South America, mainly due to the high repro-
ductive rate and migratory behavior of the species, which has
increased the number of accidents involving bees (Ferreira et al.,
2012).

The toxic effect of bee venom is usually associated with allergic
reactions, but massive attacks can be lethal due to the large amount
of venom collectively injected into the victim. Bee venom is
composed of various allergenic and pharmacologically active sub-
stances, including melittin (50e60% of the venom dry weight) and
phospholipase A2 (PA2) (11%e12% of the venom dry weight)
(Brochetto-Braga et al., 2006). Other substances are present in
smaller quantities but play an important role in the toxicity of the
venom, including apamin (2% of the venom dry weight), peptide
401 (1e2% of the venom dry weight), hyaluronidase (2% of the
venom dry weight), histamine, dopamine, adrenaline, norepi-
nephrine, and serotonin appear in lesser amounts, but play an
important role in the toxicity of venom (Han et al., 2000).

Among the birds belonging to the family Psittacidae, nine

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E.L. Milbradt et al. / Toxicon 136 (2017) 1e52
species are currently classified as members of the genus Ara
(Clements et al., 2013), including the blue-and-yellow (A. ararauna)
and red-and-green macaws (A. chloropterus), both of which are
frugivorous and relatively large with long lifespans (Sick, 2001).
A. ararauna is found in tropical regions in Central America, Brazil,
Bolivia and Paraguay, where it inhabits floodplains with buriti palm
groves, swamps, savannas and wetlands (IUCN, 2016; Jupter and
Parr, 1998; Sick, 2001). A. chloropterus is found in Brazil and other
South American countries in forested and dry terrain (Jupter and
Parr, 1998; Sick, 2001). The two species are already extinct in
some regions, while their populations are declining due to
shrinking habitat, hunting, and illegal wildlife trafficking in others
(IUCN, 2016; Jupter and Parr, 1998).

This article describes a case of a massive attack by Africanized
honeybees with a focus on the clinical and pathological aspects of
three and five specimens of A. chloropterus and A. ararauna,
respectively.

2. Case report

Three adult birds of the species A. chloropterus (mean weight of
1.5 kg) and five adults of the species A. ararauna (meanweight of 1.3
kg) were treated at the Wildlife Medicine and Research Center
(Centro de Medicina e Pesquisa em Animais Selvagens e CEMPAS)
at the School of Veterinary Medicine and Animal Science (Facul-
dade de Medicina Veterin�aria e Zootecnica e FMVZ e UNESP) in
Botucatu, S~ao Paulo, Brazil after suffering a massive attack by
honeybees.

The birds were from a conservation breeding facility located in
the state of S~ao Paulo, where they lived in 4-foot by 6-foot outdoor
enclosures surrounded by a screen of 2-inch galvanized wire. There
was no history of recent diseases or veterinary procedures, so the
diagnosis was based on the description of the attack, the observa-
tion of stingers during the clinical examination, and lesions
consistent with toxicosis caused by bee venomduring the necropsy.

The A. chloropterus birds presented swollen puncture lesions
with stingers (mainly in the facial regions without feathers),
swelling of the eyelids and subcutaneous tissue, and respiratory
distress. They were treated with 1.67 mg/kg of promethazine
(Pamergan®) and 10 mg/kg of hydrocortisone (Ariscorten®) by IM
injection followed by removal of the stingers (an average of 45
stings/bird). Approximately ten hours after receiving medication,
the birds showed alleviation of the respiratory distress, and they
could feed and drink water normally the following day. Two days
after the start of treatment, the birds showed no clinical signs and
were discharged from veterinary care.

The five A. ararauna birds were dead on arrival at the veterinary
hospital and were sent to the Laboratory of Ornithopathology
(FMVZ/S~ao Paulo State University e UNESP) for necropsy, and the
pathological changes observed during the procedure were similar
among all the examined birds (Fig. 1A). The external examination
found a large number of stingers (an average of 63 per bird) located
mainly in the non-feathered areas of the face that were associated
with erythema and edema, and food content from the crop was
found in the oral cavity and the tracheal lumen (Fig. 1B).

Examination of the subcutaneous tissue showed multifocal he-
matomas associated with the insertion of stingers and diffuse he-
matomas, and exposure of the coelomic cavity revealed marked
congestion in the heart, liver, spleen, lungs, and kidneys. Moreover,
hemorrhagic enteritis was observed, and when the skull cap was
removed, congestion was observed in the brain and cerebellum
(Fig. 1C).

Lung, heart, liver, kidney, skeletal muscle, and brain tissue
samples were fixed in 10% formalin, embedded in paraffin, and
processed for hematoxylin-eosin staining.
The lungs of all the birds (5/5) displayed swelling, congestion,
and severe bleeding. Three birds (3/5) were found to have a large
quantity of aspirated food in the lungs as well as moderate infiltrate
composed of heterophils, lymphocytes, and eosinophils (Fig. 1D).

The cardiac muscle tissue of all birds (5/5) showed significant
swelling of the myocytes, mainly in the endocardium, and vascular
dilation with erythroid repletion resulting in diffuse congestion.
The hepatic tissue of three birds (3/5) showed multifocal areas of
centrilobular necrosis associated with severe congestion and he-
patic hemorrhaging; the hepatocytes showed a high degree of
hydropic-vacuolar degeneration.

The kidneys of all birds (5/5) showed hydropic-vacuolar degen-
eration and acute tubular necrosis, which were associated with
pyknosis, karyorrhexis, and karyolysis of the nuclei of tubular cells,
cytoplasmic microvacuolation, vascular dilation, and erythroid
repletion.

Histology of the skeletal muscle tissue showed intense infiltrate
composed of heterophils, lymphocytes, and eosinophils in the
skeletal muscle fibers in a multifocal arrangement. The sarcomeres
located near the inflammatory infiltrate were swollen and degen-
erated (Fig. 1E).

The brains of four birds (4/5) showed severe congestion and
vasogenic edema, malacia, severe gliosis, and satellitosis. There was
also edema and neuropil vacuolation associated with severe
vascular congestion.

3. Discussion

The morphometric identification of the type of bee responsible
for the attack on the birds was hampered by the absence of samples
for identification, but the ferocity of the attacks and the predomi-
nance of Africanized honeybees in Brazil strongly suggest that this
strain was responsible.

Africanized honeybees attack in swarms, are ready to stingmuch
faster and with much less provocation, and are more persistent in
their attacks than their European counterparts (Tunget and Clark,
1993). This behavior is evidenced in the attack on the macaws,
since the swarm invaded the bird enclosure without any sign that
they had been provoked by the birds; this aggression has earned
Africanized honeybees the nickname “killer bees” in some countries
(Kim and Oguro, 1999). Like humans (de Oliveira et al., 2000) and
domestic animals (Oliveira et al., 2007),wild animalsmayalso suffer
massive attacks by Africanized honeybees, although no reports
involving parrots were known before the present case.

A. ararauna have blue dorsal feathers and yellow ventral
feathers as well as rows of black feathers in their facial and neck
regions (Forshaw, 2010; Jupter and Parr, 1998), and A. chloropterus
(Sick, 2001) have red feathers, green wings, and white faces with
narrow red lines (Jupter and Parr, 1998). Although both species
were in the same enclosure, the A. ararauna birds suffered a greater
number of stings that were concentrated in the facial region
without feathers. According to Vetter et al. (1999), Africanized
honeybees tend to attack the head and neck because, according to
Fitzgerald and Flood (2006), these regions are more exposed as the
bodies of most animals are covered with feathers or fur. It is known
that bee attacks can be triggered by provocation, strong odors, and
(in particular) dark colors, and moreover, when bees sting, they
release chemicals from their glands at the time and location of the
sting, which recruits and directs other bees in the swarm to the
same victim (Sherman, 1995).

The clinical signs presented in humans and animals attacked by
Africanized honeybees can vary from an allergic reaction to sys-
temic toxicity depending on the number of stings received and the
sensitivity and weight of the victim (Cardoso et al., 2003). Deaths
from multiple stings can result via three main mechanisms: direct



Fig. 1. A). Ara ararauna specimens. Note the blisters on the non-feathered facial regions and the regurgitated food adhering to the beak. B). Face of an Ara ararauna specimen. Note
the large number of stingers in the face (arrows) and facial blisters. C). Brain and cerebellum of Ara ararauna. Note the severe congestion of the blood vessels (arrows). D).
Photomicrograph of the lung of Ara ararauna. Note the intra-alveolar hemorrhage, aspirated food (short arrow), and infiltrate of heterophils (long arrow). HE. Lens: 40� magni-
fication. E). Photomicrograph of the skeletal muscle of Ara ararauna. Note the eosinophilic inflammatory infiltrate (short arrow) and the swelling of the muscle fibers (long arrow).
HE. Lens: 40� magnification.

E.L. Milbradt et al. / Toxicon 136 (2017) 1e5 3
toxicity of the venom, intravascular hemolysis caused by melittin,
or profound hypotension resulting from the inoculation and release
of endogenous histamine. Together, these mechanisms have a cu-
mulative, cascading effect that can result in multiple organ failure
represented by acute tubular necrosis and kidney failure, respira-
tory failure, rhabdomyolysis, myoglobinemia, damage to myocar-
dial cells, hepatocellular necrosis, disseminated intravascular
coagulation, and bleeding (Medeiros, 2003).

The affected birds were in adulthood with body weights be-
tween 1.3 kg (A. ararauna) and 1.5 kg (A. chloropterus), and the
A. ararauna specimens had a greater number of stingers (approxi-
mately 63/bird) than the A. chloropterus specimens (approximately
45/bird). In humans, the estimated lethal dose is approximately 500
stings per individual or 19 stings/kg body weight (Schumacher
et al., 1992; Vetter et al., 1999). The lethal dose for birds, espe-
cially those of the Psittacidae family, has not yet been determined
but seems to exceed the estimated dose for humans since the birds
that survived had received an average of 30 stings/kg of body
weight. The clinical signs presented by A. chloropterus birds, which
showed localized swelling at the sites of the stings and respiratory
difficulty, may be characteristic of an acute toxic reaction to the
large quantity of injected venom. According toWalker et al. (2005),
dogs that exhibited systemic toxic reactions to bee attacks pre-
sented vomiting, diarrhea, and respiratory difficulty associated
with acute respiratory distress syndrome.

Rahman et al. (2015) reported a massive bee (Apis mellifera)
attack on 16 pigeons (Columba livia), of which nine were adults and
seven were squabs. Twenty hours after the incident, only six adult
birds survived, and they were subsequently treated with prednis-
olone sodium succinate (10 mg/kg, IM), diphenhydramine hydro-
chloride (5 mg/kg, IM), and sulfadimidine sodium (0.2 mg/kg, IM).
The clinical signs presented by the pigeons that survived were
similar to those presented by the macaws, namely, palpebral
edema, congestion of the conjunctiva, generalized edema, edema
and erythema around the stingers, and cervical rigidity (Rahman
et al., 2015). According to these authors, five of the six surviving
pigeons died 12 hours after the start of treatment, and the number
of stingers found in each was 4, 5, 9, 10, and 11.

Honeybee venom is composed of enzymes, large peptides, and
proteins including melittin, PA2, apamin, peptide 401, hyaluroni-
dase, histamine, dopamine, adrenalin norepinephrine, serotonin,
and norepinephrine (Han et al., 2000). The swelling and congestion
found in organs during necropsymay result from the toxic effects of
the different venom components, including both the vasoactive
action of histamine and leukotrienes (Habermann, 1972) and the
tissue damage caused by melittin and PA2 (Prado et al., 2010).

In this study, the lungs of all birds (5/5) showed pulmonary
edema, diffuse congestion, and hemorrhaging; the congestion and
pulmonary edema may be associated with the large amount of
histamine injected with the venom or may be the result of
endogenous stimulus by other venom components(Schmidt, 1995).
These findings, which are characteristic of acute respiratory distress
syndrome, are also common in cases where dogs (Oliveira et al.,
2007) and humans (França et al., 1994) have suffered massive
attack by honeybees (Walker et al., 2005). Here, three birds (3/5)
presented contents in the lung parenchyma, which may indicate
that the birds suffered seizures during the agonal period that may
have been due to the action of the venom components, particularly
apamin, a neurotoxic polypeptide that can produce erratic invol-
untary movements culminating in seizures in high doses
(Habermann, 1972).

The cardiac muscle tissue was also affected by the venom,
showing necrosis, diffuse congestion, and myocardial infarction.
According to (Mjoy, 1988), the melittin, PA2, and histamine in the
venom stimulate endogenous secretion of adrenaline and norepi-
nephrine,whichmaycausemyocardial ischemia. Studies conducted
in rats have demonstrated the direct toxic action of honeybee
venom on the heart muscle, which can cause acute myocardial



E.L. Milbradt et al. / Toxicon 136 (2017) 1e54
infarction (Ferreira et al., 1995, 1994). It is believed that PA2 is
responsible for activating arachidonic acid metabolism, during
which leukotrienes and prostaglandins are released, and these
mediators may be responsible for endothelial relaxation, coronary
artery spasms, or myocardial infarction. Moreover, the venom sac
contains approximately 2 mg of histamine (Schumacher and Egen,
1995); considering the average number of stings and the weight
of the birds, each is estimated to have received, on average, 96 mg/kg
of histamine. In humans, it is known that 1 mg/kg of histamine can
produce serious cardiac disorders (Schumacher and Egen, 1995).

As in the heart, severemyositis in the skeletal muscles can result
from the action of melittin and PA2, which, alone or in synergy,
damage biological membranes by hydrolyzing phospholipids, thus
inducing pore formation and tissue lysis. Moreover, this may cause
hypercontraction and clumping of myofibrils followed by rupture,
lysis, and possible infiltration of phagocytic cells (Ownby et al.,
1997). Azevedo-Marques et al. (1992) reported that Africanized
honeybee venom induced myonecrosis in vivo in rats.

Damage to the hepatocytes is one of the consequences of
massive wasp and hornet attacks (Weizman et al., 1985), and three
birds in this study showed centrilobular necrosis associated with
severe congestion and hepatic hemorrhaging. Similar findings are
common in bee accidents involving humans (França et al., 1994),
but no change has been observed in the livers of dogs that have
suffered massive bee attacks (Oliveira et al., 2007).

Acute tubular necrosis is a commonly reported result of
poisoning by massive honeybee attacks (Grisotto et al., 2006). The
kidneys of four of the five (4/5) macaws displayed hydropic-
vacuolar degeneration and acute tubular necrosis, which may be
associated with several venom components. Melittin can compro-
mise renal blood flow in different ways: by damaging the vascular
endothelium (Schumacher and Egen, 1995), by causing vasocon-
striction (Koyama et al., 2002), by stimulating smooth muscle
contraction (França et al., 1994), by increasing the secretion of renin
(Churchill et al., 1990) and catecholamines (França et al., 1994), and
by releasing arachidonic acid (Levine and Hassid, 1977). Addition-
ally, PA2 is related to renal vasoconstriction (Imig and Deichmann,
1997) and can reduce renal blood flow by stimulating the endog-
enous release of catecholamines and vasoconstrictors (Grisotto
et al., 2006). Histamine may also reduce the rate of renal blood
flow, causing severe hypotension and acute kidney injury (Grisotto
et al., 2006). Catecholamines, such as norepinephrine and dopa-
mine, whether endogenous or from venom, can also alter renal
hemodynamics and cause hypertension ((Bourgain et al., 1998;
Bresolin et al., 2002; França et al., 1994; Munoz-Arizpe et al.,
1992; Sert et al., 1993). Therefore, kidney damage may be second-
ary; that is, it may be caused by the presence of large amounts of
myoglobin released by the rhabdomyolysis triggered by melittin
(Ownby et al., 1997; Zager, 1996).

The changes observed in the central nervous system of the
necropsied birds differed from those recorded in dogs, which show
only vascular congestion. However, an experimental study reported
neurotoxic action from bee venom in mice (Habermann and
Fischer, 1979; Oliveira et al., 2007).

4. Conclusion

The clinical signs and pathological findings observed in the birds
that suffered a massive attack from honeybees are similar to those
found in mammals and pigeons.

References

Almeida, R.A.M.D.B., Olivo, T.E.T., Mendes, R.P., Barraviera, S.R.C.S., Souza, L.D.R.,
Martins, J.G., Hashimoto, M., Fabris, V.E., Ferreira Junior, R.S., Barraviera, B., 2011.
Africanized honeybee stings: how to treat them. Rev. Soc. Bras. Med. Trop.
http://dx.doi.org/10.1590/S0037-86822011000600020.

Azevedo-Marques, M.M., Ferreira, D.B., Costa, R.S., 1992. Rhabdomyonecrosis
experimentally induced in Wistar rats by Africanized bee venom. Toxicon 30,
344e348. http://dx.doi.org/10.1016/0041-0101(92)90875-6.

Bourgain, C., Pauti, M.D., Fillastre, J.P., Godin, M., François, A., Leroy, J.P., Droy, J.M.,
Klotz, F., 1998. Massive poisoning by African bee stings. Press. Med. 27,
1099e1101.

Bresolin, N.L., Carvalho, F.L.C., Goes, J.E.C., Fernandes, V.R., Barotto, A.M., 2002. Acute
renal failure following massive attack by Africanized bee stings. Pediatr.
Nephrol. 17, 625e627. http://dx.doi.org/10.1007/s00467-002-0888-0.

Brochetto-Braga, M.R., Sergio Palma, M., Cano Carmona, E., Chaud-Netto, J.,
Rodrigues, A., Da Silva, G.P., 2006. Influence of the collection methodology on
the apis mellifera venom composition: peptide analysis. Sociobiology 759e770.

Cardoso, J.L.C., França, F.O.S., Wen, F.H., M�alaque, C.M.S., Haddad Jr., V., 2003. Ani-
mais peçonhentos no Brasil: biologia, clínica e terapêutica dos acidentes. Rev.
Inst. Med. Trop. Sao Paulo 45, S87. http://dx.doi.org/10.1590/S0036-
46652003000600009.

Churchill, P.C., Rossi, N.F., Churchill, M.C., Ellis, V.R., 1990. Effect of melittin on renin
and prostaglandin E2 release from rat renal cortical slices. J. Physiol. 428,
233e241.

Clarke, K.E., Rinderer, T.E., Franck, P., Quezada-Eu�an, J.G., Oldroyd, B.P., 2002. The
africanization of honeybees (Apis Mellifera L.) of the yucatan: a study of a
massive hybridization event across time. Evol. (N. Y) 56, 1462e1474. http://
dx.doi.org/10.1111/j.0014-3820.2002.tb01458.x.

Clements, J.F., Schulenberg, S.T., Iliff, M.J., Sullivan, B.L., Wood, C.L., Roberson, D.,
2013. The EBird/Clements Checklist of Birds of the World: Version 6.8. EBird/
Clements Checkl. Birds World Version 6.8.

de Oliveira, F.A., Guimar~aes, J.V., dos Reis, M.A., Teixeira, V.P., 2000. Acidente
humano por picadas de abelhas africanizadas. Rev. Soc. Bras. Med. Trop. 33,
403e405. http://dx.doi.org/10.1590/S0037-86822000000400012.

Ferreira, D.B., Costa, R.S., De Oliveira, J.A.M., Muccillo, G., 1995. An infarct-like
myocardial lesion experimentally induced in Wistar rats with Africanized bee
venom. J. Pathol. 177, 95e102. http://dx.doi.org/10.1002/path.1711770114.

Ferreira, D.B., Costa, R.S., Oliveira, J.S., Muccillo, G., 1994. Cardiac noradrenaline in
experimental rat envenomation with Africanized bee venom. Exp. Toxicol.
Pathol. 45, 507e511. http://dx.doi.org/10.1016/S0940-2993(11)80516-6.

Ferreira, R.S., Almeida, R.A.M.B., Barraviera, S.R.C.S., Barraviera, B., 2012. Historical
perspective and human consequences of africanized bee stings in the americas.
J. Toxicol. Environ. Heal. Part B 15, 97e108. http://dx.doi.org/10.1080/
10937404.2012.645141.

Fitzgerald, K.T., Flood, A.A., 2006. Hymenoptera stings. Clin. Tech. Small Anim. Pract.
21, 194e204. http://dx.doi.org/10.1053/j.ctsap.2006.10.002.

Forshaw, J., 2010. Parrots of theWorld, first ed. Princeton University Press, Princeton.
October 17, 2010.

França, F.O.S., Benvenuti, L.A., Fan, H.W., Santos, D.R.D.O.S., Hain, S., Picchi-
Martins, F., Cardoso, J.L.C., Kamiguti, A., Theakston, R., Warrell, D., 1994. Severe
and fatal mass attacks by “killer” bees (Africanized honey beesdApis mellifera
scutellata) in Brazil: clinicopathological studies with measurement of serum
venom concentrations. Q. J. Med. 87, 269e282. http://dx.doi.org/10.1093/
oxfordjournals.qjmed.a068927.

Funayama, J.C., Pucca, M.B., Roncolato, E.C., Bertolini, T.B., Campos, L.B., Barbosa, J.E.,
2012. Production of human antibody fragments binding to melittin and phos-
pholipase A2 in africanised bee venom: minimising venom toxicity. Basic Clin.
Pharmacol. Toxicol. 110, 290e297. http://dx.doi.org/10.1111/j.1742-
7843.2011.00821.x.

Grisotto, L.S.D., Mendes, G.E., Castro, I., Baptista, M.A.S.F., Alves, V.A., Yu, L.,
Burdmann, E.A., 2006. Mechanisms of bee venom-induced acute renal failure.
Toxicon 48, 44e54. http://dx.doi.org/10.1016/j.toxicon.2006.04.016.

Habermann, E., 1972. Bee and wasp venoms. Science 177, 314e322. http://
dx.doi.org/10.1126/science.177.4046.314.

Habermann, K., Fischer, E., 1979. Bee venom neurotoxin (apamin): iodine labeliing
and characterization of binding sites. Eur. J. Biochem. 94, 355e364.

Han, H.J., Lee, J.H., Park, S.H., Choi, H.J., Yang, I.S., Mar, W.C., Kang, S.K., Lee, H.J.,
2000. Effect of bee venom and its melittin on apical transporters of renal
proximal tubule cells. Kidney Blood Press Res. 23, 393e399 doi:kbr23393 [pii].

Imig, J.D., Deichmann, P.C., 1997. Afferent arteriolar responses to ANG II involve
activation of PLA2 and modulation by lipoxygenase and P-450 pathways. Am. J.
Physiol. Ren. Physiol. 273, 274e282.

IUCN, 2016. IUCN red list of threatened species. Version 2016-1. www.iucnredlist.
org.

Jupter, T., Parr, M., 1998. A Guide of the Parrots of the World, p. 584.
Kim, K.T., Oguro, J., 1999. Update on the status of Africanized honey bees in the

western states. West. J. Med. 170, 220e222.
Koyama, N., Hirata, K., Hori, K., Dan, K., Yokota, T., 2002. Biphasic vasomotor reflex

responses of the hand skin following intradermal injection of melittin into the
forearm skin. Eur. J. Pain 6, 447e453. http://dx.doi.org/10.1016/S1090-3801(02)
00029-0.

Levine, L., Hassid, A., 1977. Effects of phorbol-12,13-diesters on prostaglandin pro-
duction and phospholipase activity in canine kidney (MDCK) cells. Biochem.
Biophys. Res. Commun. 79, 477e484. http://dx.doi.org/10.1016/0006-291X(77)
90182-6.

Medeiros, C.R., 2003. Himen�opteros de importância m�edica. In: Cardoso, J.L.C.,
França, F.O., Wen, F.H., Sant�ana M�alaque, C.M., V.J.H. (Eds.), Animais Peçon-
hentos No Brasil: Biologia, Clínica E Terapêutica Dos Acidentes. S~ao Paulo,

http://dx.doi.org/10.1590/S0037-86822011000600020
http://dx.doi.org/10.1016/0041-0101(92)90875-6
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref3
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref3
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref3
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref3
http://dx.doi.org/10.1007/s00467-002-0888-0
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref5
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref5
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref5
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref5
http://dx.doi.org/10.1590/S0036-46652003000600009
http://dx.doi.org/10.1590/S0036-46652003000600009
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref7
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref7
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref7
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref7
http://dx.doi.org/10.1111/j.0014-3820.2002.tb01458.x
http://dx.doi.org/10.1111/j.0014-3820.2002.tb01458.x
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref9
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref9
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref9
http://dx.doi.org/10.1590/S0037-86822000000400012
http://dx.doi.org/10.1002/path.1711770114
http://dx.doi.org/10.1016/S0940-2993(11)80516-6
http://dx.doi.org/10.1080/10937404.2012.645141
http://dx.doi.org/10.1080/10937404.2012.645141
http://dx.doi.org/10.1053/j.ctsap.2006.10.002
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref15
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref15
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref15
http://dx.doi.org/10.1093/oxfordjournals.qjmed.a068927
http://dx.doi.org/10.1093/oxfordjournals.qjmed.a068927
http://dx.doi.org/10.1111/j.1742-7843.2011.00821.x
http://dx.doi.org/10.1111/j.1742-7843.2011.00821.x
http://dx.doi.org/10.1016/j.toxicon.2006.04.016
http://dx.doi.org/10.1126/science.177.4046.314
http://dx.doi.org/10.1126/science.177.4046.314
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref20
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref20
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref20
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref21
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref21
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref21
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref21
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref22
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref22
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref22
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref22
http://www.iucnredlist.org
http://www.iucnredlist.org
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref24
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref25
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref25
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref25
http://dx.doi.org/10.1016/S1090-3801(02)00029-0
http://dx.doi.org/10.1016/S1090-3801(02)00029-0
http://dx.doi.org/10.1016/0006-291X(77)90182-6
http://dx.doi.org/10.1016/0006-291X(77)90182-6
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref28
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref28
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref28
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref28
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref28
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref28
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref28
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref28
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref28
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref28


E.L. Milbradt et al. / Toxicon 136 (2017) 1e5 5
pp. 253e258.
Mjoy, E.J., 1988. Acute myocardial infarction after a wasp sting. Heart 59, 506e508.

http://dx.doi.org/10.1136/hrt.59.4.506.
Munoz-Arizpe, R.L., Valencia-Espinoza, L., Vel�asquez-Jones, L., Abarca-Franco, C.,

Gamboa-Marrufo, J., Valencia-Mayoral, P., 1992. Africanized bee stings and
pathogenesis of acute renal failure. Nephron 478. http://dx.doi.org/10.1159/
000186975.

Oliveira, E.C., Pedroso, P.M.O., Meirelles, A.E.W.B., Pescador, C.A., Gouvêa, A.S.,
Driemeier, D., 2007. Pathological findings in dogs after multiple Africanized bee
stings. Toxicon 49, 1214e1218. http://dx.doi.org/10.1016/j.toxicon.2007.01.020.

Ownby, C.L., Powell, J.R., Jiang, M.S., Fletcher, J.E., 1997. Melittin and phospholipase
A2 from bee (Apis mellifera) venom cause necrosis of murine skeletal muscle
in vivo. Toxicon 35, 67e80. http://dx.doi.org/10.1016/S0041-0101(96)00078-5.

Prado, M., Solano-Trejos, G., Lomonte, B., 2010. Acute physiopathological effects of
honeybee (Apis mellifera) envenoming by subcutaneous route in a mouse
model. Toxicon 56, 1007e1017. http://dx.doi.org/10.1016/j.toxicon.2010.07.005.

Rahman, M.M., Lee, S.-J., Kim, G.-B., Yang, D.K., Alam, M.M., Kim, S.-J., 2015. An
accidental fatal attack on domestic pigeons by honey bees in Bangladesh. J. Vet.
Med. Sci. 77, 1489e1493. http://dx.doi.org/10.1292/jvms.15-0183.

Schmidt, J.O., 1995. Toxinology of venoms from the honeybee genus Apis. Toxicon
33, 917e927. http://dx.doi.org/10.1016/0041-0101(95)00011-A.

Schumacher, M.J., Egen, N.B., 1995. Significance of africanized bees for public health.
Arch. Intern. Med. 155, 2038e2043. http://dx.doi.org/10.1001/archinte.1995.
00430190022003.
Schumacher, M.J., Schmidt, J.O., Egen, N.B., Dillon, K.A., 1992. Biochemical variability

of venoms from individual European and Africanized honeybees (Apis melli-
fera). J. Allergy Clin. Immunol. 90, 59e65. http://dx.doi.org/10.1016/S0091-
6749(06)80011-4.

Sert, M., Tetiker, T., Paydas, S., 1993. Rhabdomyolysis and acute renal failure due to
honeybee stings as an uncommon cause. Nephron 647.

Sherman, R.A., 1995. What physicians should know about Africanized honeybees.
West. J. Med. 163, 541e546. http://dx.doi.org/10.1016/S0190-9622(97)80334-9.

Sick, H., 2001. Ornitologia Brasileira.
Tunget, C.L., Clark, R.F., 1993. Invasion of the “killer” bees. Separating fact from

fiction. Postgrad. Med. 94, 92e94-98-102.
Vetter, R.S., Visscher, P.K., Camazine, S., 1999. Mass envenomations by honey bees

and wasps. West. J. Med. 170, 223e227.
Walker, T., Tidwell, A.S., Rozanski, E.A., Delaforcade, A., Hoffman, A.M., 2005. Im-

aging diagnosis: acute lung injury following massive bee envenomation in a
dog. Vet. Radiol. Ultrasound 46, 300e303. http://dx.doi.org/10.1111/j.1740-
8261.2005.00054.x.

Weizman, Z.V.I., Mussafi, H., Ishay, J.S., Shvil, Y., Goitein, K., Livni, N.,
Deckelbaum, R.J., 1985. Multiple Hornet Stings with Features Reye ’ S Syndrome
of 1407e1410.

Zager, R.A., 1996. Rhabdomyolysis and myohemoglobinuric acute renal failure.
Kidney Int. 49, 314e326. http://dx.doi.org/10.1038/ki.1996.48.

http://refhub.elsevier.com/S0041-0101(17)30187-3/sref28
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref28
http://dx.doi.org/10.1136/hrt.59.4.506
http://dx.doi.org/10.1159/000186975
http://dx.doi.org/10.1159/000186975
http://dx.doi.org/10.1016/j.toxicon.2007.01.020
http://dx.doi.org/10.1016/S0041-0101(96)00078-5
http://dx.doi.org/10.1016/j.toxicon.2010.07.005
http://dx.doi.org/10.1292/jvms.15-0183
http://dx.doi.org/10.1016/0041-0101(95)00011-A
http://dx.doi.org/10.1001/archinte.1995.00430190022003
http://dx.doi.org/10.1001/archinte.1995.00430190022003
http://dx.doi.org/10.1016/S0091-6749(06)80011-4
http://dx.doi.org/10.1016/S0091-6749(06)80011-4
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref38
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref38
http://dx.doi.org/10.1016/S0190-9622(97)80334-9
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref40
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref41
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref41
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref41
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref42
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref42
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref42
http://dx.doi.org/10.1111/j.1740-8261.2005.00054.x
http://dx.doi.org/10.1111/j.1740-8261.2005.00054.x
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref44
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref44
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref44
http://refhub.elsevier.com/S0041-0101(17)30187-3/sref44
http://dx.doi.org/10.1038/ki.1996.48

	Massive attack of honeybee on macaws (Ara ararauna and Ara chloropterus) in Brazil – A case report
	1. Introduction
	2. Case report
	3. Discussion
	4. Conclusion
	References