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LIGHT MICROSCOPIC OBSERVATIONS OF THE
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MESENTERO-PROCTODEAL REGIONS IN ADULT
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CATOPSILLA POMONA (Fabricius, 1758)
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(LEPIDOPTERA: PIERIDAE)
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Running head: Light microscopic observations of the mesentero-proctodeal
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regions in Catopsilla pomana (Fabricius, 1758) (Lipidoptera: Pieridae)
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Pisit Poolprasert1,*, Ezra Mongkolchaichana1, Sinlapachai
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Senarat2, Jes Kettratad2, Watiporn Yenchum3 and Waristha
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Angsirijinda4
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Faculty of Science and Technology, Pibulsongkram Rajabhat University, Phitsanulok,
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65000, Thailand. E-mail: poolprasert_p@psru.ac.th
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Department of Marine Science, Faculty of Science, Chulalongkorn University, Bangkok,
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10330, Thailand.
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Bio-Analysis Laboratory, Department of Chemical Metrology and Biometry,
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National Institute of Metrology (Thailand), Pathum Thani, 10120, Thailand
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Department of Veterinary Medicine, Mahanakorn University of Technology, Bangkok,
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10530, Thailand
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* Corresponding author
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Abstract
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The aim of this study was to examine the basic histological structure of
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mesentero-proctodeal regions (mid and hindguts) of Catopsilia pomona
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(Fabricius, 1758) which were living in their natural habitat in the northern
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part of Thailand. All specimens were processed by standard histological
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technique. Each section was cut at six micrometers and stained with Harris's
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haematoxylin and eosin (H&E). For the longitudinal sections, the results
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revealed that the structural wall in both mid and hindguts could be obviously
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divided into four layers: mucosa, submucosa, muscularis and serosa,
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respectively. The mid gut was mainly shown to be two types of epithelial cell,
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based on histological localization: (i) simple columnar and (ii) simple
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cuboidal epitheliums. The hindgut was distinctly classified into two subparts:
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ilium and rectum. Ilium was simple squamous epithelium, whereas the
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rectum was simple squamous epithelium with 5-6 rectal papillaeThe results
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of this study showed details of the histology of mesentero-proctodeal regions
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which are considered the first report in Thailand.
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Keywords: Catopsilia pomona, Mesentero-proctodeal regions, Histology,
Thailand
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Introduction
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The lemon emigrant Catopsilia pomona (Fabricius, 1758) is a common species of
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butterfly in Thailand, belonging to the family Pieridae (order Lepidoptera).
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Recently, C. pomona has become a serious pest of several economic plants, e.g.,
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Cassia siamea, C. fistula, C. baderiana, C. alata, C. tora, Bauhimia spp., Butea
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monosperma, Sesbania grandiflora (FAO, 2007). It is also a foliage feeder which
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prefers blossoms and young nurseries of host plants (Fabricius, 1775; Ek-Amnuay,
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2006; Raju et al., 2012; Roy et al., 2014). From the literature reviews, about the
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biological studies of C. pomona in Thailand and other countries has been
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relatively under reported.
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The several researches of digestive tract have been reported in both the
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biology and ecology of adult insects (Burgess, 1880; Bordas, 1920; Pyle, 1940;
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Mortimer, 1965; Chauthani and Callahan, 1967; Le Grice, 1968; Judy and Gilbert,
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1970; Beals and Berberet, 1976; Gongalves, 1981). Based upon histological
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studies, some lepidopterans have been mainly examined such as Diacrisia obliqua
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(Kabir and Ameen, 1986), Papilio demoleus (Goyle, 1990) and Papilio polytes
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(Gaikwad et al., 2011). Kabir and Ameen (1986) and Goyle (1990) described that
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the mesentero-proctodeal regions has been well documented with the structure
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particular due to containing special cell types as the absorptive function.
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Therefore, investigation about normal histology of this organ will provided basic
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knowledge to apply for the nutritional physiology as well as agricultural pest
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management. However, investigation of the mesentero-proctodeal regions (mid
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and hindguts) in C. pomona (Fabricius, 1758) has not been reported. In this
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research, we examined the C. Pomona mesentero-proctodeal region of subjects
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inhabiting natural habitat in northern part of the Thailand.
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Materials and Methods
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Twenty-five pooled samples of adult C. pomona (total length about 5-7
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cm) were gathered from Lampang, Phitsanulok and Chiang Mai provinces,
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northern part of Thailand during October-December 2013. The whole abdomen of
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each butterfly was mainly dissected and then immediately fixed in Davidson's
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fixative for 36-48 hours. Each specimen was processed using standard histological
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techniques (Bancroft and Gamble, 2002). As to detail, the specimens were
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dehydrated in serially graded ethanol, clearing in xylene and were infiltrated in
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paraffin (m.p., 55-60 ̊C). The paraffin block was cut at 5-6 µm by rotary
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microtome and stained with Harris's haematoxylin and eosin (H&E). Afterwards,
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observations of histological structure were done under a light microscope.
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Results and Discussion
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Digestive tract of C. Pomona was divided to three parts by considering the
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structure of the tissues, including fore, mid and hindguts. This study reports only
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on the midgut and hindgut of the C. Pomona. In longitudinal sections, wherein the
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two walls of their regions could be distinctly divided into four layers: mucosa,
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submucosa, muscularis and serosa, respectively.
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Histological observations of C. pomona midgut
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The midgut was a long and elongate tube (Fig. 1) which began from the
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esophageal or stomodeal valves to the pyloric valve. Based on histological
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sections, the cardiac valve protruded into the hindgut. It consisted of two cell
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layers; inner and outer (Fig. 1A). In the outer cell layer, the epithelial cells were
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elongated and high simple columnar with predominant oval nucleus as basophilic.
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The cytoplasm was slightly stained as acidophilic, whereas the inner cell layer
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was simple cuboidal epithelium which presented a large round or spherical
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nucleus, along the length of the mid gut. Only epithelium cells lined the basement
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membrane and it was rarely surrounded by connective tissue (Fig. 1A-1B). Then,
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these epitheliums gradually increased in cell into simple columnar epithelium.
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The functional structure of the cardiac valve prevents the return of food
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(Lacombe, 1971).
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As to detail, histological structure of midgut showed as follows (Fig.
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1C-1D); mucosa layer: the mucosa was raised to lumen in longitudinal folds
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(100-120 µm). Moreover, it was constituted of two types of epithelium cells based
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on the histological characterization and localization. The upper longitudinal fold
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was lined with a simple columnar epithelium (about 15-20 µm). It presented the
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microvilli and was supported by a prominent basement membrane. The nuclei of
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epithelium cells were oval with prominent nucleolus as strongly basophilic
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stained. In the lower longitudinal fold, single-layered cuboidal epithelium was
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observed. Its rounded or oval nucleus with scattering chromatin was observed.
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Submucosa layer: this layer was slightly comprised of the thin layer of loose
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connective tissue. Muscularis layer: it was rarely constituted the thin layer of
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smooth muscle. Serosa: the histological structures were simple squamous
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epithelium.
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In addition, Malpighian tubules were observed between hindgut and
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midgut (Fig. 2A-B). Each Malpighian tubule showed a single layer of cuboidal
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epithelium. The nucleus of tubule was oval with scatting chromatin. Its cytoplasm
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also showed the acidophilic stained. Externally, it was surrounded by loose
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connective tissue. This characterization was similar to research on various insects
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including Lepidoptera (Standlea and Yonke, 1968; Mathur, 1972), Coleoptera
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(Areekul, 1957) and Hymenoptera (Caetano and Overal, 1984; Arab and Caetano,
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2002).
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Histological observations of Catopsilia pomona hindgut
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The hindgut had different histological structures from the mid gut. In the
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last part of mid gut a longitudinal fold was not present but it was jointed and
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changed into tall simple columnar epithelium. The histological structure between
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mid and hindguts also had a pyloric valve (Fig. 2A). The function of this structure
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was to move food into hindgut (Gillott, 1995).
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Generally, this organ was mainly divided into two subparts (Fig. 2B-C):
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Ileum and rectum. As to detail, ileum had a smaller lumen. Its overall histological
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structure was observed, the mucosa was slightly extended into the lumen and little
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branch. Epithelial cells were flat and poorly developed as simple squamous
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epithelium with covering by cuticle. This finding differs from some Lepidoptera,
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Papillio polytes polytes (Gaikwad et al., 2011) and P. demoleus which is
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composed of a single of cuboidal epithelial cells (Goyle, 1990). Submucosa layer,
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it rarely consisted of thin layer connective tissue. Muscularis layer was externally
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surrounded by smooth muscle in both outer circular and inner longitudinal muscle
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layers. The outer layer was serosa showing thin connective tissue. The function of
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this region concerns food digestion and nutrient absorption (Irvine et al., 1988;
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Schumaker et al., 1993; Serrão and Cruz-Landim, 1996; Serrão et al., 2004). For
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rectum, the mucosa layer extended more into the lumen with 5-6 rectal papillae.
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This result differs from Micropteryx spp. and Hepialus spp. with 3-4 rectal
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papillae (Mortimer, 1965) and 30-35 rectal papillae in Hyalophora cecropia (Judy
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and Gilbert, 1970). Each rectal papilla was a single layer of simple squamous
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epithelium and also presented oval nucleus. The epithelium was covered with
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cuticle. Other layers were similar with the prior region. The function of rectum is
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osmotic control (Serrão and Cruz-Landim, 1996; Serrão et al., 2004). In addition,
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Cruz-Landim (1985) reported that it also demonstrated water and nutrient
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absorption.
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Conclusions
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According to this research, we can conclude that the structural wall in
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mesentero-proctodeal regions could be distinctly separated into four layers;
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mucosa, submucosa, muscularis and serosa, respectively. The mid gut was lined
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by two types of epithelial cell (simple columnar and simple cuboidal epitheliums),
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whereas the hindgut was lined by simple squamous epithelium in both ileum and
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rectum. Other parts of the hindgut were similar to that described in the mid gut.
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This information here offered a basic knowledge for further development at the
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immunocytochemistry and ultrastructural level.
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Acknowledgements
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The authors would like to thanks Dr. Angoon Lewvanich for her help in
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taxonomic study and the members of the Fish Research Unit, Department of
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Pathobiology, Faculty of Science, Mahidol University, Bangkok and Biology
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Program, Faculty of Science and Technology, Pibulsongkram Rajabhat
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University, Phitsanulok for their technical support in laboratory. We also thank
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Dr. David V. Furman for critically reading the manuscript.
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Figure 1. Micrograph of midgut of C. pomona (A-C). CT = connective tissue,
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Ev = Esophageal valve, Fg = foregut, Ocl = Outer cell layer, Icl = Inner cell
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layer, Mg = Midgut, M = Mucosa, Sm = Submucosa, Mc = Muscularis, Msc =
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Simple columnar epithelium, Msce = Simple cuboidal epithelium.
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Figure 2. Micrograph of hindgut of C. pomona; (A-C). G = Ganglia, Hg =
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Hindgut, I = Ileum, Pv = Pyloric valve, M = Mucosa, Mc = Muscularis, Mg =
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Midgut, Mt = malphigian tubule, R = Rectum, Rp = Rectal papillae, S =
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Serosa.