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Ultrastructure of adhesive device in fly in

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Article in Parasitology Research · May 2006

DOI: 10.1007/s00436-005-0100-0 · Source: PubMed

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Parasitol Res (2006) 98: 477–481
DOI 10.1007/s00436-005-0100-0
ORIGINA L PA PER
K. L. Sukontason . N. Bunchu . R. Methanitikorn .
T. Chaiwong . B. Kuntalue . K. Sukontason
Ultrastructure of adhesive device in fly in families calliphoridae,
muscidae and sarcophagidae, and their implication as mechanical
carriers of pathogens
Received: 20 November 2005 / Accepted: 28 November 2005 / Published online: 14 January 2006
# Springer-Verlag 2006
Abstract The ultrastructure of adhesive device or the
pulvilli, pad-like structure between the tarsal claws of the
legs, is presented in the blowfly (Calliphoridae), housefly
and relatives (Muscidae), and flesh fly (Sarcophagidae)
through scanning electron microscopy. The blowfly
species were Chrysomya chani, Chrysomya nigripes,
Chrysomya pinguis, and Chrysomya villeneuvi; while
those of the housefly and relatives were Musca domestica
and Hydrotaea chalcogaster, respectively. Fresh fly
species included Boettcherisca peregrina and Liosarco-
phaga dux. Numerous tenent setae were observed on the
ventral side of the pulvilli. Two features of the tip of the
tenent setae were found as a spatula-like (in C. chani, C.
pinguis, C. nigripes, M. domestica, H. chalcogaster, B.
peregrina, and L. dux) and spoon-like tip (in C.
villeneuvi). Transmission electron microscopy of the
tenent setae in M. domestica revealed the electron-lucent
centrally located, suggesting an adhesive substance. These
results provided anatomical information that allow us to
not only understand the successful attachment of flies to
smooth surfaces but also clarify their role as a mechanical
carrier of microorganisms.
Introduction
Several species of flies in the families Calliphoridae
(blowflies), Muscidae (housefly and its relative), and
K. L. Sukontason (*) . N. Bunchu . R. Methanitikorn .
T. Chaiwong . K. Sukontason
Department of Parasitology, Faculty of Medicine,
Chiang Mai University,
Chiang Mai 50200, Thailand
e-mail: klikitvo@mail.med.cmu.ac.th
Tel.: +66-53-945342
Fax: +66-53-217144
B. Kuntalue
Electron Microscopy Research and Service Center (EMRSC),
Faculty of Science, Chiang Mai University,
Chiang Mai 50200, Thailand
Sarcophagidae (flesh flies) have been incriminated as a
public health concern in many parts of the world. Due to
their life cycle of synanthropic behavior, which is closely
associated with the human environment, along with their
feeding habit, adult flies in these groups could be
mechanical carriers of numerous pathogenic microorgan-
isms from filth to human food, thereby causing diseases
to humans (e.g., Greenberg 1971; Levine and Levine
1991; Sukontason et al. 2000; Sulaiman et al. 2000).
These pathogens (bacteria, virus, protozoa cyst, helminth
ova, or larva) have been mechanically carried by flies via
the external integument, particularly organs such as the
mouth parts, wings, legs, or adhesive device that come
into contact with filth (Greenberg 1973; Tan et al. 1997).
Study using scanning electron microscopy (SEM)
revealed that a large number of bacteria, Escherichia
coli 0157:H7, adhered to the surface of housefly mouth
parts and proliferated in the minute spaces of the
labellum, thereby suggesting a none simple mechanical
carrier (Kobayashi et al. 1999). The adhesive device of
the terminal structure between the tarsal claws of flies has
also been demonstrated through SEM as a mechanical
transporter of microorganisms (Greenberg 1973). In this
publication, we present the ultrastructure of the adhesive
device in eight fly species, commonly found in down-
town Chiang Mai, north Thailand, to reveal the func-
tional morphology of this organ as a mechanical carrier of
microorganisms.
Materials and methods
Three fly species used in this study were from laboratory
colonies maintained at the Department of Parasitology,
Faculty of Medicine, Chiang Mai University, Thailand.
They belonged to the family Calliphoridae (Chrysomya
nigripes), Muscidae (Musca domestica), and Sarcophagi-
dae (Liosarcophaga dux). The other five species were
obtained from a fly collection in Chiang Mai Province,
northern Thailand. They belonged to the family Call-
iphoridae (Chrysomya chani, Chrysomya pinguis, and
478

Fig. 1 SEM of the adhesive device of flies. Top view of the Fig. 4 SEM of the adhesive device of flies. Ventral view of the
tarsomere of the blowfly, C. chani, showing the tarsal claw (CL) and tarsomere of fresh fly B. peregrina showing the densely packed
pulvilli (PU) setae of the pulvilli (PU) and empodium (EP)

Fig. 2 SEM of the adhesive device of flies. Ventro-oblique view of Fig. 5 SEM of the tenent setae on the ventral surface of fly pulvilli.
the tarsomere of the housefly M. domestica showing the tarsal claw Spatula-like tip of the tenent setae of C. chani
and pulvilli. Arrow indicates small attached material

Fig. 3 SEM of the adhesive device of flies. Top view of the

tarsomere of the fresh fly B. peregrina showing the tarsal claw (CL) Fig. 6 SEM of the tenent setae on the ventral surface of fly pulvilli.
and pulvilli (PU) Spatula-like tip of the tenent setae of C. pinguis
 479

Fig. 7 SEM of the tenent setae on the ventral surface of fly pulvilli. Fig. 10 SEM of the tenent setae on the ventral surface of fly
Spatula-like tip of the tenent setae of C. nigripes pulvilli. Spatula-like tip of tenent setae of H. chalcogaster

Fig. 8 SEM of the tenent setae on the ventral surface of fly pulvilli. Fig. 11 SEM of the tenent setae on the ventral surface of fly
Spoon-like tip of the tenent setae of C. villeneuvi pulvilli. Spatula-like tip of tenent setae of B. peregrina

Fig. 9 SEM of the tenent setae on the ventral surface of fly pulvilli. Fig. 12 SEM of the tenent setae on the ventral surface of fly
Tenent setae of M. domestica showing spatula-like and pointed-like tip pulvilli. Spatula-like tip of tenent setae of L. dux
480
Fig. 13 Ultrathin cross section through the tenent setae of the hind
legs of M. domestica. Serial position of the tenent setae
Chrysomya villeneuvi), Muscidae (Hydrotaea chalcoga-
ster), and Sarcophagidae (Boettcherisca peregrina).
Adult flies of each species were killed by placing them in a
plastic bag put in a freezer that is set at 4°C for approximately
30 min. Flies killed individually were prepared for SEM
processing, as described by Sukontason et al. (2005). Only
M. domestica was selected as a subject model for inspection
of the ultrathin section using transmission electron micros-
copy (TEM); the procedure of Sukontason et al. (2005) was
followed.
Results
Basically, each fly leg was composed of a coxa, trochanter,
femur, tibia, and five tarsal segments. Dorsally viewed, the
pretarsus of the blowfly C. chani consisted of a pair of claws
with sharp apices and a pair of pulvilli underneath each claw,
by which, the empodium and unguitractor plate were not
clearly seen (Fig. 1). For the housefly M. domestica,
numerous tenent setae were seen on the ventral side of the
pulvilli and claws (Fig. 2). In the sarcophagid B. peregrina,
the dorsal view of the pulvilli represented a remarkably large,
Fig. 14 Ultrathin cross section through the tenent setae of the hind
legs of M. domestica. Higher magnification of tenent seta showing
an electron-dense thick wall, with the central electron-lucent
smooth pad (Fig. 3), while the ventral surface illustrated
heavily packed tenent setae (Fig. 4).
In the examination using higher magnification of the
pulvilli, the terminal setae of C. chani (Fig. 5), C. pinguis
(Fig. 6), and C. nigripes (Fig. 7) were as morphologically bent
outward as a spatula-like tip; however, those of C. villeneuvi
were as bent inwardly as a spoon-like tip (Fig. 8). The terminal
setae of M. domestica, H. chalcogaster, B. peregrina, and L.
dux were similar to spatula-like tips (Figs. 9, 10, 11, and 12,
respectively). However, some pointed tips were observed in
M. domestica (Fig. 9).
An ultrathin cross section through the tenent setae of M.
domestica revealed an electron-dense thick wall, in contrast
with the central electron-lucent (Figs. 13 and 14).
Discussion
The ultrastructure of the adhesive device has been investi-
gated using SEM in several insect groups, such as Coleoptera
(Betz 1996, 2003; Betz and Mumm 2001), Hymenoptera
(Federle et al. 2001; Federle and Endlein 2004), Strepsiptera
(Pohl and Beutel 2004), Diptera (McAlpine 1991; Gorb
1998; Niederegger et al. 2002; Niederegger and Gorb 2003;
Gaume et al. 2004), and Heteroptera (Gorb and Gorb 2004).
In insects, the adhesive device is not only used principally in
locomotion and attachment on a smooth surface but also in
catching and as a defense against predators (Betz and Mumm
2001; Betz and Kölsch 2004; Gorb 2004). In the blowfly
Calliphora vicina, the contact behavior role of the tenent
setae in attachment has been revealed using a high-speed
video recording (Niederegger et al. 2002; Niederegger and
Gorb 2003).
In this study, the SEM investigation showed two remark-
able features of the tip of the tenent setae: the spatula-like tip
(as seen in C. chani, C. pinguis, C. nigripes, M. domestica, H.
chalcogaster, B. peregrina, and L. dux) and spoon-like tip (as
seen in C. villeneuvi). The spatula-like tip in this study was
similar to type I setae in the syrphid fly Episyrphus balteatus
(Syrphidae) (Gorb 1998), kelp fly Amma blancheae (Coelo-
pidae) (McAlpine 1991), or blowfly Calliphora vimitoria
(Gaume et al. 2004). However, either the bearing spatula-like
or spoon-like tip of the tenent setae enabled flies to increase
the number of contact points for attachment to a surface (Gorb
and Gorb 2004).
The electron-lucent at the central region of the tenent setae
of M. domestica, as seen in TEM, was consistent with E.
balteatus (Gorb 1998). It has been suggested that the
electron-lucent areas may be due to lipid-containing sub-
stances, most of which are dissolved during the TEM
procedure (Gorb 1998). Investigation in Stenus (Coleoptera:
Staphylinidae) showed that the beetle appears to release
tarsal secretion through the tenent setae, within which lipid
and proteinaceous fraction are contained (Betz 2003). In C.
vicina, the tenent setae were reported as filled with secretion
(Gorb 1998). Likewise, an adhesive fluid was secreted by the
fly pad of C. vomitoria (Gaume et al. 2004). Investigation
on another blowfly species Calliphora erythrocephala
showed that the location of the secretion released opened at

the end of the tenent setae (Gorb 1998). Regarding this, the
electron-lucent area in the tenent setae of M. domestica
would imply that the substance secreted through their tip.
Such substances appeared to be important not only for the
successful attachment to smooth surfaces (Gorb 2004), but,
in view of the mechanical vector, also as a glue for
attaching microorganisms (see Fig. 2, arrow).
No remarkable differences in the microtopography of the
adhesive device were observed between sexes of the fly
species examined. These similarities would, therefore, be
involved by similar functions and/or roles displayed by
each adult sex. Investigations in northern Thailand showed
that there was no difference between either sex of adult M.
domestica in the role played as a mechanical carrier of 42
bacterial species; for example, coagulase-negative staphy-
lococci, E. coli, Viridans streptococci, nonfermentative
gram-negative bacilli, Klebsiella pneunoniae, Morganella
morganii, Enterobacter cloacae, Providencia stuartii, Pro-
teus mirabilis, Aeromonas sobria, Aeromonas hydrophila,
Pseudomonas aeruginosa, Vibrio cholerae non-01, and
Staphylococcus aureus (Sukontason et al., unpublished data).
In conclusion, this investigation revealed the morpholog-
ical features of adhesive devices in flies in the families
Calliphoridae, Muscidae, and Sarcophagidae, which are
commonly found in Thailand. These results provide ana-
tomical information that allows us to clarify the role of these
flies as a mechanical carrier of many microorganisms.
Acknowledgement This work received support from the Faculty of
Medicine Endowment Fund for Medical Research.
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