CIIAPTER-2
Characterization of hemolymph agglutinins in
centipedes
Abstract
Different species of centipedes belonging to order Scolopendroniorpha and
Jeophilomorpha were collected from different locations of Tamilnadu and Kerala,
ndia. Species belonging to Scolopendromorpha were the most commonly available
entipedes. The naturally occurring agglutinin activity in the hemolymph of
entipedes was assessed by hemagglutination assay (HA). All the species of
entipedes examined showed HA activity but the HA profile of each species was
ifferent. The different species of centipede surveyed for HA activity agglutinated
thbit erythrocytes much better than all other tested erythrocytes. Based on
magglutination (HA) titer, the species can be ranked as Rhysida nuda nuda
1ewport) > Rhysida longipes longipes (Newport) >
Scolopendra amazonica
ucherl) > Otostigmus nudus (Pocock) > Olostig,nus ceylonicus
(Hasse) >
tostigmus sps. > Sepedonophilus antiodus (Pocock). Since the common garden
ntipede Rhysida nuda nuda (Newport) showed a remarkably high HA titer, this was
:lected as a model for further characterization of the agglutinin. Among the various
;sues tested for HA, the titer obtained with hemolymph was the highest. Physical
rameters such as body length, weight and sex of the centipede had no influence on
e HA titer of the hemolymph.
Introduction
Noguchi (1903), first observed the presence of hemagglutinins in the
lomic fluid of the horseshoe crab Limulus polyphemus, an arachnid and in
55
Hornarus americanus, a crustacean. Since then a number of investigators purified and
characterized the agglutinins in the limulids of North America Limulus polyphemus
(Cohen et al., 1965, 1972; Watne et al., 1966; Cohen, 1968; Marchalonjs and
Edelman, 1968; Finstad et al., 1972; Roche and Monsigny, 1974; Oppenheim
et al.,
1974; Roche et al., 1975, 1977; Nowak and Barondes, 1975; Sela et al.,
1975; Kaplan
t al., 1977; Maget Dana et al., 1975; Robey and Liu, 1981), Japan
Tachypleus
rridentatus (Shimizu et al.,
1977, 1979; Shishikura and Sekiguchi, 1983, 1984;
Yamaguchi 1987; Fischer et al., 1994; Okino et al., 1995; Saito et al., 1995,
1997)
md India Carcinoscorpius rotundacauda (Bishayee and Dorai, 1980; Dorai
et al.,
982 a, 1982 b; Mohan et al., 1982; Srimal et al., 1985). Lectin from hemolymph of
imulids agglutinated both mammalian erythrocytes and bacteria (Pistole, 1976, 1978;
4c Sweegan and Pistole, 1982; Brandin and Pistole, 1983). Since the purified
gglutinins or lectins of limulidae were specific for sialic acids, they were
ommercially made available for diagnosis of sialoglyco-conjugates of tumor tissues
d bacterial pathogens. Agglutinins found in the hemolymph of arachnids were also
)und to be sialic acid-specific (Cohen et al., 1979; Brahmi and Cooper, 1980; Vasta
d Cohen, 1982, 1983, 1984; Vasta and Marchalonis, 1983; Ahmed et al., 1986;
hoang etal., 2001). The origin and source of the hemolymph lectin of arachnids are
)t known.
Another class of arthropod that has been extensively investigated for
molymph agglutinins are crustaceans (Cantacuzene, 1912,1919; Tyler and Metz,
)44,1945; Tyler and Scheer, 1945; Tyler, 1946; Faglioni et al., 1971; Miller et al.,
p 72; Comick and Stewart, 1973; Pauley, 1974 a, 1974 b; Hall and Rowland, 1974 a,
56
1974 b; Ghidalja et al., 1975; Hartman et
al., 1978; Kamiya and Ogata, 1982; Vasta
et al., 1983; Ravindranath and Cooper, 1984; Ravindranath and Paulson, 1987;
Kamiya et al., 1987; Ratanapo and Chulavatnatol, 1990; Umetsu
et al., 1991; Vargas-
Albores et al., 1992; Muramoto et al.,
1991, 1994, 1995; Mercy and Ravindranath,
1992, 1993; Muralj et al., 1994; Nalini et
al., 1993; Fragkiadakis and Stratakis,1995;
Zenteno et al., 2000; Comjnettj et al., 2002; Juan et al., 2005).
Class Insecta also has been extensively surveyed for agglutinins (Glaser,
1919; Gary et al., 1948; Bemheimer,1952; Briggs, 1958; Feir and Walz, 1964;
Vlarek, 1970; Scott, 1971; Anderson et al.,1 972;
Amirante, 1976; Amirante and
4azzalaj, 1978; Amirante et al., 1976; Komano et al.,
1980; Lackie, 1981; Flapner
nd Jermyn, 1981; Mauchamp, 1982; Hapner, 1983; Suzuki and Natori, 1983; Ingram
t al., 1983; Umetsu et al., 1984, 1993; Stebbins and Hapner,
1985; Stynen et al.,
985; Kitagaki etal., 1986; Pendland and Boucias, 1986, 1993; Qu
etal., 1987; Kubo
id Natori, 1987; Bellah et al., 1988; Ammanai et al., 1990; El Deeb
et al., 1990;
rubhoffer and Matha, 1991; Mc Kenzie and Preston, 1992; Flirayama et al., 1993,
)94; VoIf et al., 1995; Haq et al.,1996; Basseri, 2004; Jayalakshmi,
et al., 2005).
sect hemolymph also contains agglutinins binding to sialic acids, with diversity in
gar specificity. It is not clear whether any particular organ or tissue is specifically
volved in the production of the agglutinins.
Among myriapoda only millipedes had been studied by Basil-Rose (1999),
d it is suggested that midgut may be the site of synthesis of the agglutinin.
In spite of extensive investigation on the agglutinins of arachnids, crustaceans,
d insects, very little attempt had been made to identify and characterize the
57
agglutinin or lectin of class myriapoda. Although phylogenetically myriapoda are
closely related to insects (Tiegs and Manton, 1958), the perusal of literature showed
that very few studies had been undertaken to compare these closely related phyletic
groups for the characteristics of the agglutinins. Our objective was to characterize
hemolymph agglutinins and to identify its source in order to understand the biological
role of the agglutinins in the pathobiology and physiology of centipedes.
Fhe specific aims of this chapter are
1. To identify the presence of the hemolymph agglutinin in centipedes.
. To identify the possible source of hemolymph agglutinin in centipedes by
screening the tissues for the presence of agglutinins.
To select a particular species of centipede expressing a high level of HA titer that
it can be purified and characterized.
Materials and methods
Seven species of centipedes, Rhysida nuda nuda (Newport), Rhysida longipes
)ngipes (Newport) , Scolopendra amazonica (Bucherl), Otostiginus nudus (Pocock),
tostigmus ceylonicus (Hasse) , Otostigmus sps. , Sepedonophilus antipodus
ocock) were collected according to the availability from districts of Kerala and
amilnadu. They were maintained individually in separate plastic containers
)ntaining moist mud and decaying leaves and fed with insect larva, termites and
)ckroach. Acclimation of centipedes to the laboratory condition and breeding them
as difficult because of their cannibalistic nature.
58
Hemolymph collection
Hemolymph was obtained by cutting the hind leg off initially and then cutting
a leg close to the body and carefully squeezing the animal or by cuffing the antennae.
Before handling, the centipedes were narcotized with chloroform. Care should be
;aken while pressing to avoid contamination with gut contents (Willie et al., 1990).
he hemolymph was collected into eppendorf tubes kept on ice and stored in the
efrigerator. The hemolymph was clot free, and remained as such without any
rariation in I-IA titer for more than three months.
p reparation of tissue extract
Healthy centipedes were dissected and the tissues were removed and
Loroughly rinsed in cold Tris buffered saline (TBS) to remove the hemolymph. 100
g each of foregut, midgut, hindgut, fat body, ovary, nerve cord, malpighian tubule
id muscle were collected, ground well in a glass tissue grinder, suspended in Imi
'cold TBS and centrifuged for 10 mm. at 4000 g and the supernatant was assessed
r hemagglutinating activity.
paration of hemocytes
The hemocytes were separated using the method of Soderhall and Smith
83). The leg or antennae of the centipede was cut and the hemolymph was
Ilected in 1.35 ml of ice-cold anticoagulant buffer (citrate-EDTA : trisodium citrate
mM) and citric acid (26 mM) in 1:1 ratio by volume (200 ml), in 50 ml of NaCl
59
(71 MM), glucose (100 MM), and EDTA disodium salt (10 mM) at 4 0 C). The
mixture of hemolymph and buffer was shaken gently to assist rapid mixing and
centrifuged (200 x g, 2 mm. at 4 0C). The hemocyte pellet was resuspended in
1.5 ml
of iso-osmotic buffer (Tris-HCI (50 mM), NaCl (150 mM), and CaC1 2
(1 mM) pH
7.5).
N
Erythrocyte collection
Erythrocytes from several mammals were collected for hemagglutination
issay. Blood for this purpose was obtained by heart puncture (rat), or venipuncture of
he ear (rabbit), fore arm (man) or neck (horse, donkey, cow, buffalo, sheep, and
oat) or from the slaughter house (pig). Erythrocytes were collected directly in
riodified Alsevier's medium containing sodium citrate (30 mM, pH 7.1), sodium
hloride (77 mM), glucose (114 mM), neomycin sulfate (100 mg/ml), and
hloramphenicol (330 mg/ml). They were suspended and washed three times with
n volumes of Iris-buffered saline TBS, pH 7.5 (Tris-HC1 50 mM, NaCI 100 mM,
aC12 10 mM) and resuspended in the same as 1.5 % suspension.
[emagglutination assay
Hemagglutination assays were carried out as described by Ravindranath and
iulson (1987). Briefly, hemolymph and tissue extracts (25 jil) were serially diluted
ith 25 t1 of TBS in 'U' shaped microtiter wells (Rigid polystyrene, NUNC) and
ixed with 25 jil of 1.5 % erythrocyte suspension. The plates were incubated for one
on
hour at 30°C. The hemagglutination titer or HA titer (the units of agglutinin activity)
is the reciprocal of the highest dilution of the sample that gave agglutination.
Biochemical analysis
Water content
Known quantity of hemolymph or tissues was dried in a desiccator. The
lifference between the wet weight and dry weight gave the amount of water present
n the tissue (Passoneau and Williams, 1953; Mullainathan, 1979).
alcium content
Hemolymph calcium was measured following O-Cresolpthalein complexone
ethod (Calcium kit). Calcium in alkaline medium reacts with O-Cresolpthalein
omplexone to . from a purple coloured complex whose absorbance was proportional
calcium concentration. The interference due to magnesium and iron was
iminated by using 8-hydroxy quinoline. To the test tube of blank, standard and test
Id working reagent, calcium standard and serum respectively. Mix well and allow
em to stand at room temperature for 5 minutes. Measure the absorbance of
indard and test against blank on a spectrophotometer at 570 nm within 30 mm.
;timation of protein
The protein concentration was estimated by Folin-Ciocaltue method (Lowry
al., 1951). The principle of this method has two steps (1) The carbamyl group of
)tein molecules reacts with copper and potassium of the biuret reagent to give a
61
blue colored copper-potassium biuret complex (2) This complex together with
tyrosine and phenolic compounds present in the protein reduce the phosphomolybdate
of the Folin reagent to intensify the colour of the solution.
Ethanolic precipitate of 50 j.il of hemolymph was dissolved in 1 N NaOH: Jo
this 5 ml of reagent mixture (50 t1 of reagent A: 2 g of Na2 (CO3
) in 100 ml of 0.1 N
NaOH and 1 ml of reagent B: 500 mg of cupric solution in 1 % sodium or potassium
tartarate) was added and mixed well for 10 minutes. Then 0.5 ml of Folin phenol
reagent was added and mixed rapidly and after 30 minutes absorbancy was measured
at 500 nm.
Results
Distribution of centipede species in and around Kanyakumari
Seven species of centipedes Rhysida nuda nuda (Newport), Rhysida
(ongpes longzpes (Newport), Scolopendra amazonica (Bucherl), Otostigmus nudus
Pocock), Otostigmus ceylonicus (Hasse), Otostigmus sps and
Sepedonophilus
intipodus (Pocock) representing the class Chilopoda were collected from different
;ites of Kerala and Tamilnadu. The systematic account and distinguishing
norphological characteristics of different species are provided (Fig. 2.1-2.7). The
ite and season of collection are given in Table 2.1.
ollection of centipedes
Centipedes remain under stones or in burrows in the soil during summer
eason. Soon after monsoon they emerge out of their habitat and spread on the
mounding vegetation. They were collected after rain and brought to the laboratory
62
for study. Scolopendra amazonica (Bucherl), Otostigmus nudus
(Pocock),
Otostiginus ceylonicus (Hasse), Otostigmus sps. , and
Sepedonophilus antipodus
(Pocock) were found in rubber estates and Rhysida nuda nuda
(Newport), Rhysida
longipes longipes (Newport) were commonly found in gardens under the pots. It
was observed that use of pesticide in gardens gradually reduced the population of
centipedes. The animals were collected from various sites to survey the presence of
lectins.
Hemolymph agglutination in centipedes
Agglutinins are found in the hemolymph of Rhysida nuda nuda
(Newport),
?hysida longipes longipes (Newp ort), Scolopendra amazonica (Bucherl),
rudus (Pocock), Otostigmus ceylonicus (Hasse), Otostigmus sps. and
Otostigmus
Sepedonophilus
'ntipodus (Pocock). The HA titer differed among the seven species tested. The
bility of hemolymph to agglutinate erythrocyte of different mammalian species and
e agglutination titer are presented in Table 2.2.
lemagglutinating activity in tissues
of Rhysida nuda nuda
Rabbit erythrocytes were used as indicator cells in detecting agglutinin
tivity in tissue extracts of Rhysida nuda nuda. The agglutinin activity was
Dserved in almost all the tissues tested with varying HA titer (Table 2.3, Fig 2.8).
owever, the HA activity in the hemolymph was higher than the tested tissue
tracts.
IN
Thus it can be conveniently deduced, that agglutinin activity was high in the
hemolymph and it was ideal for further study.
Biochemical and Biological factors in relation to HA activity
Variation in weight and size and their respective biochemical constitution
such as water, protein and calcium content of the hemolymph, had no influence on
HA activity of hemolymph agglutinin of Rhysida nuda nuda (Table.2.4, 2.5)
Discussion
Centipedes are an economically important group of arthropods playing
ialuable role in the control of noxious pests in the terrestrial ecosystem (Yadav,
994). They particularly of the members of the family scolopendridae are often
reated as creatures of nuisance value owing to their poisonous nature and painful
ite. Geophilomorph centipedes are commonly known as earthiovers due to their
urrowing activity and habitat preferences. The microhabitat of all the centipedes is
e wet and moist areas, and they inhabit the environs below rotten barks, dry foliage,
tones, heap of cowdung etc. In captivity they feed on insect larva, cockroaches,
ioths and other insects, worms etc.
A survey of the agglutinins in the hemolymph of different chilopods by
ialyzing its hemagglutinating activity (HA) revealed the presence of agglutinins.
he agglutinins showed differential affinity with different species of erythrocytes.
he inability of the hemolymph agglutinin to agglutinate erythrocytes of some
ammalian species suggested that these erythrocyte membranes may express
fferent types of cell surface receptors which were not recognized by the agglutinins.
64
Chilopod agglutinins though capable of agglutinating a variety of erythrocytes, they
have specific affinity for rabbit erythrocytes that contain NeuGc and 9-0-acetyl
NeuAc on its glycocalyx (Pfeil et al., 1980). Rabbit erythrocyte specific lectin was
also reported in the hemolymph of millipedes, a myriapodan representative (Basil
Rose, 1999). Similar results were obtained with insect Extatosoma tiaratum
(Richards and Ratcliffe, 1990), Melanoplus sanguinzpes (Jurenka et al.,
1982),
Locusta migratoria (Drif and Brehelein, 1989) and Periplaneta americana
(Mercy
and Basil, 1991), Phiebotomus dubosqui (Voif et al., 2002), Anopheles stephensi
Basseri et al., 2004) and arachnids,
Carcinoscorpius rotundacauda (Dorai et
ii., 1982).
Among the various chilopodan hemolymph surveyed for HA Rhysida nuda
uda showed high HA titer with rabbit erythrocytes. In addition to hemolymph
gglutinin, agglutinin with ability to agglutinate rabbit erythrocyte was also observed
i all the tissues of the centipede R. nuda nuda. Similar observations have been
ported in the hemolymph and midgut gland extract of another myriapodan species hyropygus descriptus (Basil Rose, 1999), hemolymph, midgut and hindgut of
isects Anopheles gambiae (Mohamed et al., 1992), Glossina species (Ibrahim et al.,
84; Ingram and Molyneux, 1988; Maudlin and Welbum, 1987) and in crustaceanspatopancreas (Fragkiadakis and Stratakis, 1997). However in Rhysida nuda nuda
.e HA activity of the hemolymph with rabbit erythrocyte was higher when compared
ith the other tissues analysed. This indicates that the region of agglutinin activity
as localized in the hemolymph, which attributed to its physiological function.
NN
The hemolymph HA titer for rabbit erythrocytes differed with diverse genera
of centipedes. From our observation it could be summarized that agglutinins showed
diverse erythrocyte specificity and the functions attributed to agglutinin as
recognizing molecule was reflected in its binding specificity to erythrocytes.
Evidently, the agglutinin had a specific recognition role in pathological process.
Pathogens present in the vicinity of the different species of centipedes and their
Issociation with the hosts may be reflected in the difference or similarities in the HA
;iter. The variation in the titer may also reflect the difference in the rate of
)iosynthesis or utilization of the agglutinin for clearance of the pathogens. Even after
earing the organism in the laboratory for a month, the HA remained unaltered as it
vas for the freshly collected centipedes revealing that this agglutinin was a natural
.gglutinin with predetermined receptor specificity.
Since maximum
iemagglutinability was observed in the hemolymph of the centipede Rhysida nuda
uda with rabbit erythrocytes, we have decided to analyse the immunological role of
e hemolymph lectin of R. nuda nuda in the subsequent chapter.
'uture directions
The observation reported in this chapter compels further characterization of
ie agglutinin namely the sugar specificity of hemolymph agglutinin. Once the
pecific nature of the agglutinin is understood, undoubtly the immunological role of
gglutinin in centipedes could be studied.
TV
salient findings
[) Hemolymph of the centipede contained agglutinins that preferentially agglutinated
abbit erythrocytes.
) Though all the tissues tested, contained an agglutinin capable of agglutinating
abbit erythrocytes, maximum HA titer was observed in the hemolymph.
) Of the various species of centipedes analyzed of hemagglutinating ability, Rhysida
uda nuda hemolymph showed greater agglutinability than the other species tested.
) Season plays a major role in the availability of the centipedes.
Cannibalism among the centipedes was the major problem faced in collective
aring of these animals in the laboratory.
The poisonous . bites of centipedes led to severe itching and local inflammation.
67
Table 2. 1 Time and site of collection of centipedes
Site of collection
Species
Time of collection
Place
Rhysida nuda nuda
(Newport)
Throughout the
year
Rhysida longipes
longipes (Newport)
"
Scolopendra
amazonica (Bucherl)
Otostigmus nudus
(Pocock)
Otostigmus ceylonicus
(Hasse)
Otostigmus sps.
Sepedonop hi/us
antipodus (Pocock)
Feb - March
July - September
District
Poojapura
Red fields
Samayapuram
Nagercoil
Trivandrum
Coimbatore
Trichy
Kanyakumari
Nagercoil
Kanyakumari
Table 2.2 Hemagglutination titer of hemolymph agglutinins
from different species of centipedes of order Chilopoda against
different mammalian erythrocytes
Presence of hemagglutinin was identified by agglutinating vertebrate erythrocyte with
serial dilutions of the hemolymph. Hemolymph (25 p1) was serially diluted with Trisbuffered saline (pH 7.5) and mixed with 1.5% suspensions of rabbit erythrocytes. Titer was
determined as the reciprocal of the highest dilution of hemolymph giving complete
agglutination after 60 minutes at 30° C (room temperature).
thro-
type
10)
Centipedes
Rhysida Rhysida
longipes
nuda
nuda
longipes
Scolopendra Otostigmus Otostigmus Otostigmus Sepedonophilus
ceylonicus
sps.
amazonica nudus
antipodus
512
bit
16384
128
e
128
128
64
16
16
an A
32
16
64
an B
32
0
32
32
16
64
32
16
32
32
8
8
It
16
16
32
falo
16
16
128
nan 0
4
2
128
se
4
4
8
ikey
2
0
8
V
16
8
256
8
128
2
128
4
64
4
64
4
32
2
0
0
128
2
0
0
64
2
0
0
64
2
0
0
16
16
0
0
2
0
2
0
4
0
2
2
2
2
0
0
0
0
0
0
Table 2.3 Naturally occurring agglutinin in the tissues of
Rhysida n uda n uda
25tl of tissue extracts (concentration 100 mg /ml) were serially diluted in Tris
buffered saline (pH 7.5) and mixed with 1.5% suspension of rabbit erythrocytes. Titer is
determined as the reciprocal of the highest dilution of the extract giving complete
agglutination after 60 minutes at 30° C (room temperature).
Tissue
(n=5)
HA titer
Rabbit erythrocytes
Hemolymph
16384
Hemocytes
4096
Foregut
8192
Midgut
8192
Hindgut
4096
Muscle
2048
Nerve cord
1024
Appendage
32
Cuticle
16
Table 2.4 Biochemical study of hemolymph of
Rhysida nuda nuda
The water content of the hemolymph was estimated by the method of Passoneau and
William (1953) and Mullainadhan (1979), calcium content by O-Creso!pthalein complexone
method (Webster, 1962) and protein content by Foiin-Ciocaiteu method (Lowry et al., 1951).
Characteristics analyzed
(n= 10)
Water g%
Hemoiymph
20± 0.2
Total Protein (mg/ml)
4.5 ± 0.141
Total Calcium (mM/I)
14.5 ± 0.5 12
HA titer (Rabbit erythrocytes) 16384
Table 2.5 Hemagglutination (HA) titer of the hemolymph of
Rhysida nuda nuda in relation to length, weight
and sex
Hemolymph from Rhysida nuda nuda (Newport) of different length, weight
and sex were collected and assayed for HA.
HA titer
Characteristics analyzed
(n - 5)
Rabbit erythrocytes
Length- 3-8 cm
16384
Weight - 0.9-1.6 gm
16384
Sex- Male
16384
Female
16384
Fig 2.1 Rhysida nuda nuda
(Newport)
Systematic position
Class
Order
Family
Sub family
Tribe
Genus
Species
ADULT
Chilopoda
Scolopendromorpha
Scolopendridae
Otostigminae
Otostigmini
Rhsida
nuda nuda (Newport)
t
ADULT WITH LARVA
LARVA
I
Salient Features: Colour - Cephalic plate & tergite olive green to brownish,
sternites + legs; Body length- 45mm, including antennae and oral
legs; Head- Cephalic plate wider with a median notch;
Coxosternum- Dental plate of the maxillipede with 4+4 teeth;
Tergites- smooth, complete paramedian longitudinal furrows,
only end tergite laterally emarginated, without median furrow.
Sternites: Smooth, without paramedian longituidinal furrows.
1St tarsal segment, 19t"
Legs: 2-18 pairs with 2 spurs to the
20th without spur. Anal legs two and a half times longer.
one and
Coxopleura with 2 apical spines.
Fig 2.2 Rhysida longipes longipes (Newport)
Systematic position
Class
Order
Family
Sub family
Tribe
Genus
Species
ADULT
Chilopoda
Scolopendromorpha
Scolopendridae
Otostigminae
Otostigmini
Rhysida
longipes longipes (Newport)
ADULT WITH LARVA
Pik
-
rent Features
Tergites, except anterior segments, with a pair of
paramedian longitudinal furrows; emargination of
tergites beginning from 15th segment; process of coxopleura
with a lateral spine; prefemur of anal legs with a process; 1-12
pair of walking legs with 2 spurs to the 1t tarsal segment.
Fig 2.3 Scolopendra ama2onica (Bucherl)
Systematic position
Class
Sub Class
Order
Family
Sub family
Tribe
Genus
Species
Chilopoda
Epimorpha
Scolopendromorpha
Scolopendridae
Scolopendrinae
Scolopendrini
Scolopendra
amazonica (Bucherl)
ADULT
ADULT WITH EGG
OA
-
C __
LARVA -
F!
4
PARENTAL CARE
ADULT WITH LARVA
Salient features: First tergite anteriorly overlaid by cephalic plate; a pair
of spinules at the base of the claws of last leg 20 1h pair of
walking legs without tarsal spur. Colour green or yellow
usually with a darker cross bar on each body segment,
legs yellowish orange, large and robust centipedes.
Fig 2.4
Otostigmus nudus (Pocock)
Systematic position
Class
Order
Family
Sub family
Tribe
Genus
Species
Chilopoda
Scolopendromorpha
Scolopendridae
Otostigminae
Otostigmini
Otostigmus
nudus (Pocock)
---- - :--
Salient features: Colour-Cephalic plate and whole body black;
large spiracles oval or cycle form with their
long axes facing vertically or diagonally.
Fig 2.5
Otostigmus ceylonicus (Hasse)
Systematic position
Class
:
Order
:
Family
:
Sub family :
Tribe
:
Genus
:
Species
:
Chilopoda
Scolopendromorpha
Scolopendridae
Otostigminae
Otostigmini
Otostigmus
ceylonicus (Hasse)
Salient features: Colour-Cephalic plate & whole body
black; legs with alternate white and black
bands; spiracles oval form or cycle form.
Fig 2.6
Otostigmus sps.
Systematic position
Class
:
Order
:
Family
:
Sub family :
Tribe
:
Genus
:
Species
:
Chilopoda
Scolopendromorpha
Scolopendridae
Otostigminae
Otostigmini
Otostiginus
ND
I
a
Salient features: Colour-Cephalic plate white, whole body
black; legs with alternate white and black
bands; spiracles oval form or cycle form.
Fig 2.7 Sepedonophilus antipodus (Pocock)
Systematic position
Class
Order
Family
Genus
Species
..:
.
L
F '
-
Chilopoda
Geophilomorpha
Geophilidae
Sepedonophilus
antipodus (Pocock)
k
\.
Salient features: 9-200 mm in length, 27-191 pairs of legs,
slender body of 33 or more segments, fihiform
antennae with 14 segments, no ocelli, lack
Tomosvary organs, all tergites same length,
pleurites present, final pair of legs modified
into posterior antennae, lacks tarsal claw.
Fig 2.8 Naturally occurring agglutinin in the tissues
of Rhysida nuda nuda
4. F
4
3,5
3
• hemolymph
Ell foregut
2.5
U midgut
• hindgut
• muscle
4-
2
o nervecord
• appendage
• cuticle
I
1.5
0.5
0
hemolymph
foregut
midgut
hindgut
Tissues
muscle
nervecord
appendage
cuticle