b e n i - s u e f u n i v e r s i t y j o u r n a l o f b a s i c a n d a p p l i e d s c i e n c e s 2 ( 2 0 1 3 ) 1 0 8 e1 1 3

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Full Length Article

Histopathological characterization of induced

Paederus dermatitis caused by Egyptian rove
beetles (Paederus alfierii)

Mohamed S. Ahmed a,*, Hessien A. Boraei b, Osama M. Rakha b

a
Pathology Department, Faculty of Veterinary Medicine, Kafrelsheikh University, Kafr El-Sheikh 33516, Egypt
b
Entomology Department, Faculty of Agriculture, Kafrelsheikh University, Kafr El-Sheikh 33516, Egypt

article info abstract

Article history: Outbreaks of Paederus dermatitis (PD) have been reported in Kafr El-Sheikh governorate
Received 20 January 2013 when Rove Beetles (genus Paederus) are accidentally brushed or crushed on the skin,
Accepted 10 March 2013 releasing haemolymph pederin. The aim of this study was to characterize the histopatho-
Available online 1 October 2013 logical changes of induced (PD) caused by Egyptian Rove Beetles (Paederus alfierii). Haemo-
lymph extracts (pederin) of one and three insects were applied on shaved skin of two groups
Keywords: of rats. Skin specimens were collected after 12 h, 3, 7, 14 and 30 days after the extract
Paederus dermatitis application and processed for histopathological examination. Gross changes were indicated
Pederin by erythema, eruptions, marked epidermal necrosis and large bullae formation. Micro-
Haemolymph scopically, there was initial eosinophilic infiltration followed by a rapid destruction of the
Paederus alfierii epidermis associated with exudation. The exudate penetrated the injured area of the
Skin epidermis forcing the degenerated cells apart and forming spaces which coalesced to form a
vesicle with severe infiltration of the inflammatory cells in the dermis. Vesicles then dried
forming crusts followed by repair which took a longer time in case of higher concentration.
Conclusion: Reviewing our literature, this is the first time in Egypt to study the histopath-
ological change of (PD) associated with P. alfierii. The induced dermatitis in this study is
relatively more severe compared to (PD) of other Rove Beetles occurring in other regions of
the world. The degree of dermatitis was directly correlated with the concentration of the
toxin. There was no systemic reaction of the topical application of pederin on skin.
Copyright 2013, Beni-Suef University. Production and hosting by Elsevier B.V. All rights reserved.

1. Introduction Staphylinidae, with more than 370,000 species. The dermatitis is

caused by a vesicant chemical contained in the body fluids of
Beetles are a group of insects belonging to the order Coleoptera. the beetles. The chemical in the body of both Oedemeridae and
There are three families, Oedemeridae, Meloidae and Meloidae is cantharidin. The third group of blister beetles

* Corresponding author. Tel.: þ20 1006892801, þ20 1003046551; fax: þ20 47 3 231311.
E-mail address: aosayedahmed@yahoo.com (M.S. Ahmed).
Peer review under the responsibility of Beni-Suef University

Production and hosting by Elsevier

2314-8535/$ e see front matter Copyright 2013, Beni-Suef University. Production and hosting by Elsevier B.V. All rights reserved.
http://dx.doi.org/10.1016/j.bjbas.2013.03.002
 b e n i - s u e f u n i v e r s i t y j o u r n a l o f b a s i c a n d a p p l i e d s c i e n c e s 2 ( 2 0 1 3 ) 1 0 8 e1 1 3
belongs to the genus Paederus (family Staphylinidae) which does
not bite or sting, but when crushed against the skin it releases a
potent vesicant and notorious toxin called pederin which re-
sults in itching, burning and erythema on human skin (Al-
Basheer et al., 2002; Davalos et al., 2002). The toxin is
dispersed throughout the body of beetles and is located in their
haemolymph (Qadir et al., 2006).
Although (PD) is a specific form of acute irritant contact
dermatitis caused by pederin, the mechanisms that lead to
these skin reactions are not fully understood. It would appear
that some content of the insect body fluid released by the slap
of the insect to the skin of the patient which evoked the in-
flammatory response observed. The causative insect in the
reported cases is nocturnal inhabit thus explaining the timing
of the bites (Oyedeji et al., 2006).
A major outbreak involving 2000 cases of vesicular
dermatitis was reported in Okinawa in 1966 (Armstrong and
Winfield, 1969). In Malaysia, cases were reported among 12
medical students who were residents of University Science of
Malaysia hostels in Kelantan (Mokhtar et al., 1993). An
outbreak of acute vesicating dermatitis was noted among staff
members on night shifts and some patients in the open wards
of a suburban hospital in Sri Lanka (Kamaladasa et al., 1997).
One hundred and fifty-six cases of Paederus dermatitis were
also reported among patients attending a dermatology clinic
in northern Iran from May to October 2001 (Zagari et al., 2003).
Thousands of high rise flat dwellers and dormitory students
were affected by a similar outbreak of dermatitis linearis in
Penang in 2002 (Rahmah and Norjaiza, 2008). Another
outbreak of PD occurred among 36 schoolchildren attending a
night tuition class in Malaysia (Rahmah and Norjaiza, 2008). In
Egypt, the rove beetle, Paederus alfierii, is an active predator of
several insect pests attacking a wide variety of cultivated
plants which causes necrotizing dermatitis and conjunctivitis
when left in contact with the human skin and eye respectively
(Morsy et al., 1996).
Students in the campus of Kafr El-sheikh University com-
plained from skin irritations like burns at morning without any
history of exposure to burning agents. Similar complain was also
recorded from local people, they mentioned that a marked in-
crease in P. alfierii population occurred annually during partic-
ular months (January to May) of the year and lesions like burns
were noticed on the exposed skin after crushing these insects at
the same time of insect population increase then decreased and
completely disappeared after June. Symptoms began as itching,
erythema, eruption and then vesicles appeared after few hours
to one to two days. The vesicles then oozed liquid materials
which in infected cases converted to pus for few days then dried
and formed crusts. Healing occurred nearly after one month
leaving scar in the affected areas. So, the aim of this study was to
characterize the histopathological changes of induced Paederus
dermatitis (PD) caused by the Egyptian Rove Beetles (P. alfierii).
2. Materials and methods
2.1. Beetles
Adult P. alfierii were collected from several fields in Kafr El-
Sheikh governorate for extraction of their haemolymph.
109
Beetles were ground in a small volume of water (30 ml dem-
ineralized water/insect). Extraction of the suspension pro-
ceeded with 100 ml ethyl acetate and repeated three times. The
solvent was removed from the extract in a centrifugal evap-
orator. The dry sample was again dissolved in 50 ml water and
extracted with 100 ml hexane in order to eliminate most of the
accumulated lipids, whereas pederin is left in the watery
phase (Rupert et al., 1995).
2.2. Experimental animals
Forty five adult albino rats of 80e100 g in weight were obtained
from faculty of medicine, Tanta University, Egypt. Rats were
housed in wire cages under standard conditions with free
access to drinking water and food. The rats were kept in
temperature-controlled room with 14 h Light and 10 h dark
cycles and were given standard diet and water adlibitum. The
rats were left for 2 weeks without any treatment for accli-
matization. The rats were further divided into three equal
groups after shaving small areas on their skin for application
of the insect’s haemolymph extract for only one time. In group
(A) fifteen rats were treated with an extract of only one insect,
group (B) fifteen rats were treated with extract of three insects
and control group (C) fifteen rats were treated with distilled
water.
2.3. Histopathology
Skin specimens from three rats of each group were collected
after 12 h on 3, 7, 14 and 30 days of the extract application and
fixed in 10% neutral buffered formalin. Samples were dehy-
drated by ascending grades of alcohol, cleared in xylene and
embedded in paraffin wax, mounted, sectioned at 5 mm,
stained with hematoxylin and eosin (H&E) and then examined
by light microscopy (Bancroft and Stevens, 1996).
3. Results
3.1. Gross findings
The gross changes appeared after application of the insect’s
extracts were the same for both treated groups, however these
signs were more severe and appeared earlier in the rats treated
with the extract of three insects. A rapid reaction was noticed
after 12 h of extracts application in the form of slight erythema
with redness at the site of application. After three days, the red
areas became elevated and slightly flabby. Moderate to
severely swelled skin areas were noticed after seven days of
extract application. After 14 days, the swelled areas formed
crusts and turned to be small injured to nearly healed after 30
days in the rats of group (A) while in the rats of group (B) there
were incomplete healing with crust formation (Figs. 1 & 2).
3.2. Microscopic findings
Twelve hours after the extract application, there was eosin-
ophilic dermatitis which was slight in the rats of group (A)
and moderate to severe in the rats of group (B), there were
infiltration of eosinophils and polymorphs in the dermis
110 b e n i - s u e f u n i v e r s i t y j o u r n a l o f b a s i c a n d a p p l i e d s c i e n c e s 2 ( 2 0 1 3 ) 1 0 8 e1 1 3

Fig. 1 e Photograph (group [A] after 30 days) showing Fig. 3 e Photomicrograph (group [A & B] after 12 h) showing
almost complete healing of the treated skin. slight infiltration of eosinophils {arrows} in the dermis
(H&E, 400).

(Fig. 3). After three days of application in rats of group (A)

After seven days of application in group (A) there was slight
there were slight necrotic changes of dermis with increased
necrosis of almost the whole epidermal layers leaving rem-
infiltration of eosinophils, polymorphs and lymphocytes
nants of the cellular nuclei and hyaline membranes accom-
accompanied with slight edematous changes (Fig. 4), but in
panied with infiltration of inflammatory cells in the necrotic
rats of group (B) there was a perifollicular and intrafollicular
areas and around the skin appendages with edema, while in
inflammatory infiltrate, mainly of lymphocytes accompanied
rat of group (B) there were complete epidermal necrosis and
by macrophages and granulocytes forming the image of
infiltration of large number of intra-epidermal neutrophils
“swarm of bees” (inflammatory cells around the hair follicle
forming a massive layer of necrotic tissue covering the un-
bulbs) in the dermis (Fig. 5). Marked to severe necrotic
derlying dermis (Fig. 8).
changes and edema were noticed in the epidermis with su-
After 14 days of application, in rats of group (A), there was
perficial perivascular infiltrate of lymphocytes and histio-
nearly complete healing of skin with formation of few layers
cytes. The exudate had penetrated the epidermis separating
of epidermal cells covered with sloughed necrotic scab (Fig. 9),
the necrosed epithelial cells and forming small chambers
but in rats of group (B) there were incomplete healing of the
which represented the beginning of vesicle formation. Inside
necrosed epidermis in which the neighboring healthy
the vesicle there was fluid with degenerated epithelial cells
epidermal cells propagated underneath the necrotic tissue
“ghosts”. The basal cell layer was intact or indistinct and
(Fig. 10).
there was even destruction of the dermo-epidermal junction
(Figs. 6 & 7).

Fig. 4 e Photomicrograph (group [A] after 3 days) showing

massive infiltration of eosinophils, polymorphs and
Fig. 2 e Photograph (group [B] after 30 days) showing lymphocytes {blue arrow} with edema {black arrow} in the
incomplete healing with crust formation. dermis (H&E, 400).
 b e n i - s u e f u n i v e r s i t y j o u r n a l o f b a s i c a n d a p p l i e d s c i e n c e s 2 ( 2 0 1 3 ) 1 0 8 e1 1 3 111

Fig. 5 e Photomicrograph (group [B] after 3 days) showing Fig. 7 e Photomicrograph (group [B] after 3 days) showing
perivascular, perifollicular {green arrows} and severe necrosis of epidermis {yellow arrow} with massive
intrafollicular {black arrows} inflammatory cells infiltrate inflammatory cell infiltration in the dermis {black arrow}
forming the image of “swarm of bees” (H&E, 400). and vesicle formation {blue arrow} (H&E, 400).

After 30 days of application, the rats of group (A) revealed

the allergic inflammatory responses of the skin accompanied
complete healing with formation of intact epidermis and skin
with release of some neutral serine proteases that led to
appendages with few numbers of inflammatory cells still
disruption of desmosomes, detachment of tonofilaments and
present in the dermis in and around dilated blood vessels
breakdown the homeostasis of epidermal cells allowing the
(Fig. 11). In the other hand, healing in the rats of group (B) was
exudates to enter the cells and form vesicles. This finding was
incomplete with formation of young epidermal cells covered
in agreement with Smith et al. (2002) who reported that ped-
with almost freely separated scabs (Fig. 12).
erin like cantharidin is absorbed by the lipid layers of
epidermal cell membranes and the main pathophysiological
changes observed in irritant contact dermatitis were skin-
4. Discussion
barrier disruption, epidermal cellular changes, and cytokine
release mainly from keratinocytes. Meanwhile, Davidson et al.
In the present study, local irritation and redness of skin
(2009) demonstrated the presence of histamine in the secre-
occurred after about 12 h of the insects’ extracts application
tion of some genera of beetles. Within a day, vesicles appeared
which then turned to vesicles after three days. This was due to
over the erythematous area, ruptured within few days

Fig. 6 e Photomicrograph (group [B] after 3 days) showing

severe necrosis of epidermis {yellow arrow}, degenerated Fig. 8 e Photomicrograph (group [B] after 7 days) showing
epithelial cells “ghosts” scattered in the vesicle {blue severe epidermal necrosis {star} and intense inflammatory
arrow} with destructed dermo-epidermal junction {black cells infiltration forming a massive layer of necrotic tissue
arrow} (H&E, 400). covering the underlying dermis {black arrow} (H&E, 400).
112 b e n i - s u e f u n i v e r s i t y j o u r n a l o f b a s i c a n d a p p l i e d s c i e n c e s 2 ( 2 0 1 3 ) 1 0 8 e1 1 3
Fig. 9 e Photomicrograph (group [A] after 14 days) showing
nearly complete healing of skin with formation of few
layers of immature epidermal cells {arrow} covered with
sloughed necrotic scab {star} (H&E, 200).
forming erosions or crusting or developing secondary infec-
tion (Borroni et al,. 1991, Lisa Moed, et al., 2001, Cardoso and
Haddad, 2005 and Padhi et al., 2007).
We thought that the urticating properties were due to the
secretion of an urticating liquid by cells located at the base of
hollow hairs and called trichogen cells that is in line with
Banney et al. (2000) and Wilkinson and Beck (2004) who
claimed that when pederin penetrates the skin, the irritating
liquid performs its action.
After the 3e4 days of first appearance of erythema and
eruption, vesicle formation was completed and then other
changes had followed. During the formation of the vesicle in
the epidermis, a more extensive process was found in the
Fig. 10 e Photomicrograph (group [B] after 14 days)
showing propagation of the neighboring healthy
epidermal cells {arrow} underneath the necrotic tissue
{star} with infiltration of large number of inflammatory
cells {arrow head} (H&E, 200).
Fig. 11 e Photomicrograph (group [A] after 30 days)
showing complete healing: Few inflammatory cell reaction
{black arrow} under a thin tightly formed epidermis {blue
arrow} in and around dilated dermal blood vessels {orange
arrow} (H&E, 400).
dermis. There was a perifollicular and intrafollicular inflam-
matory infiltrate, mainly of lymphocytes accompanied by
macrophages and granulocytes forming the image of “swarm
of bees” (inflammatory cells around the hair follicle bulbs).
This picture can be considered a pathognomonic feature for
the dermatitis caused by this insect. The inflammation led to
edema, microvesiculation, necrosis, and the appearance of
macrophages in the dermal papillae, parakeratosis and su-
perficial perivascular infiltrate of lymphocytes and histiocytes
are also evidence described by Khan et al. (2009).
After seven days, the edema regressed with severe
desquamation of the epidermal cells. Such desquamation has
been described earlier in a report from northern Iran (Bircher,
Fig. 12 e Photomicrograph (group [B] after 30 days)
showing incomplete healing: Formation of few layers of
epidermal cells {arrow} covered with almost freely
separated large scab {star} (H&E, 400).
 b e n i - s u e f u n i v e r s i t y j o u r n a l o f b a s i c a n d a p p l i e d s c i e n c e s 2 ( 2 0 1 3 ) 1 0 8 e1 1 3
2005). This desquamation and epidermal necrosis were prob-
ably attributed to a more potent toxin produced by the local
species of P. alfierii compared with the other species of Paede-
rus beetles in other countries. Higher potency of the toxin in
this study is also indicated by the presence of large vesicles
and necrosis in the skin of all treated rats, which is a rarely
reported finding in Paederus dermatitis (Nikbakhtzadeh and
Sadeghiani, 1999).
In this study, healing time ranged from 14e30 days in rats
treated with an extract of one insect to more than 30 days in
rats treated with an extract of three insects. This result was in
agreement with Zagari et al. (2003) who noticed that recovery
occurred within 8e10 days as the lesions dry and dark scales
appeared. After 14 days when the destructive process was at an
end, the demarcation between the living and the necrotic
epithelium became distinct. The living cells were growing
rapidly inward over the floor of the vesicle presented the same
characteristics as that growing over any defect. After four
weeks the black scales start to peel off, exposing new pink skin.
They added that in some cases the scars vanished after six
month, meanwhile; in one case scars were still apparent after
12 months. Whereas, Couppie et al. (1992) mentioned that the
uncomplicated lesions of Paederus dermatitis healed within
short time about 7e10 days. Dursteler and Nyquist (2004) also
found gradual resolution of the lesions with residual areas of
hypo- and hyper-pigmentation in the patients in Pakistan. The
acute vesicular lesions were found to undergo crusting within
few days and healed completely in 10e12 days (Davalos et al.,
2002). This finding can also be explained on the basis of a more
severe inflammatory reaction occurred against a more potent
toxin produced by the local species of beetles (P. alfierii).
It was obvious that the effect of the haemolymph extract
from three insects produced more pronounced effect than
that of one insect. The same chemical may produce different
responses depending upon its concentration, duration of
exposure, and individual responses, although; the nature,
concentration and duration of contact with a chemical are of
primary importance (Qadir et al., 2006).
‘It is worthy to mention that, there was no internal or
systemic involvement due to topical application of pederin
throughout the time of the experiment which was unlike the
findings reported by Nikbakhtzadeh and Tirgari (2008) who
recorded a new finding in only one individual who was suf-
fered from difficulty in breathing and explained that by two
possibilities. The first was that the patient was hypersensitive
to Pederin and the second that the affected site was the nose
and surrounding area leading to dyspnea. Meanwhile, no one
has reported any adverse systemic involvement with the
topical application of pederin.
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