VOLUME 5 NO. 1 JANUARY 2014 • pages 35-42 MALAYSIAN JOURNAL OF VETERINARY RESEARCH

DESCRIPTIONS ON THE MORPHOLOGY OF SOME OF THE MALAYSIAN DOMESTIC (Gallus domesticus) USING SCANNING ELECTRON MICROSCOPY

WAHAB A. RAHMAN* AND NURUL HUDA MANAP Faculty of Science and Food Technology Universiti Malaysia Trengganu, 21030 Kuala Trengganu, Malaysia. * Corresponding author: [email protected]

ABSTRACT. A total of fi ve of the can cause damage to the host, especially in nematodes, Acuaria spiralis, Gonyglonema severe infections. ingluvicola, Ascaridia galli, Heterakis Nematodes are elongated, cylindrical gallinarum and Oxyspirura mansoni were and un-segmented roundworms. They are recovered from the Malaysia domestic covered with a tough, non-cellular layer chicken, Gallus domesticus and studied called the cuticle and they have a well- for their morphology. Scanning electron developed alimentary tract. Most species microscopy was used to study and observe of nematodes are bisexual and they a single the outer surface of the nematodes. The worm (Butcher, Hogsette and Jacobs, morphological of the five species of 1997). nematodes were described and compared. In terms of size, they appear small, Keywords: morphology, nematodes, inconspicuous and seemingly unimportant Malaysian domestic chicken, scanning to humans. However, some nematodes electron microscopy can cause diseases of great importance to humans and domestic and wild plants and INTRODUCTION (Gerald and Larry, 1996) whereas some are benefi cial in attacking insect Nematodes are invertebrate roundworms pests, mostly sterilizing, or otherwise and they inhabit marine, freshwater, and debilitating their hosts (Grewall, Ehlers terrestrial environments. Nematodes and Shapiro-Ilan, 2005). According to belong to the phylum Nematoda (or Gerald and Larry (1996), nonparasitic Nemata) which includes parasites of plants nematodes may accidentally fi nd their way and of animals, including that of humans; into a vertebrate and become short-lived, they also feed on bacteria, fungi, algae and and they can become a pathogenic parasite. on other nematodes (Platt, 1994). Due to Most species are of bilateral its large number of species, nematodes symmetry (Croll, 1976). Lee (1965) stated are the most important group of helminth that the male nematode has a single parasites of poultry. These parasitic species testis and the female possesses ovarian

35 MALAYSIAN JOURNAL OF VETERINARY RESEARCH VOLUME 5 NO. 1 JANUARY 2014

tubules. Many species of nematodes are and laterally and known as the longitudinal oviparous, while some are ovoviviparous lines. Lateral lines contain the longitudinal or viviparous. The adult stage is reached canals of the excretory system. For the after 4 larval stage molts. Male nematodes muscular layer, which follows next and are small in size compared to female (Lee, lines the body cavity, consists of a number 1965). Eggs are the transmission stage; of cells having a basal contractile portion some produce the infective larvae that which is transversely striated, and a penetrate host body. Nematodes have six cytoplasmic portion which contains the stages in their life cycle; egg, four juvenile nucleus and it is also connected to the stages and adult (Croll and Matthews, nerve trunks in the dorsal or ventral line. 1977). This muscular layer is divided into four A typical nematode has a cylindrical quadrants by longitudinal lines (Soulsby, or fi lariform body with the extremities 1968). usually more or less attenuated or The present paper describes the truncated. The length varies from less morphology of fi ve species of nematodes than one millimeter to more than one in the Malaysian domestic chicken, Gallus meter, the ratio of length to breadth domesticus using scanning electron varying considerably. They have more microscopy. Morphological differences elastic and tough cuticle as compared with between the fi ve species were compared. cestodes and generally smooth, seldom spined, scaled or scattered with bosses MATERIALS AND METHODS and transversely, longitudinally or rarely obliquely striated (Yamaguti, 1963). Samples The body wall of nematodes comprise of the cuticle, hypodermis, and body wall The study was conducted on fi ve species of musculature. The outermost covering is nematode. The fi ve species were obtained the cuticle, a complex structure of great fresh from the Malaysian domestic chicken, functional significance to the animals. Gallus domesticus, obtained from various The cuticle also lines the stomodeum, farms in the northern parts of Peninsular proctodeum, excretory pore, and vagina. Malaysia. The fi ve species of nematodes A carbohydrate-rich coat, the surface i.e. Acuaria spiralis, Gonyglonema of the cuticle is found in free-living ingluvicola, Ascaridia galli, Oxyspirura and parasitic nematodes, 5 to 20 mm in mansoni and were thickness (Blaxter et al, 1992). The cuticle identifi ed as according to the descriptions is formed by an underlying sub-cuticle of Lancaster (1957), Lim (1971), Mustaffa- layer, called the hypodermis. This layer Babjee (1980) and Rahman et al. (2009). forms four longitudinal thickenings on the inner aspect, situated dorsally, ventrally

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Preservation of samples RESULTS AND OBSERVATIONS

The nematodes, both male and female From the observations using LM, Acuaria were preserved in 70% alcohol. Higher spiralis is characterised by having concentrations of alcohols were avoided ‘cordons’ (cuticular ridges or grooves) at which may result in structural dehydration the cuticle of the anterior part of the body artifacts, including shrinkage and body (Plate 1). The cordon may be non-recurrent surface distortions suffi cient to obscure or recurrent where it runs down the body features required for morphological and turn back forwards again. The lips identification and analysis, thereby are usually small and triangular in shape compromising precise morphometrics whereas the pharynx is cylindrical. Plate (Naem et al., 2010) 2 showed the anterior and posterior of the female worm. At the posterior end is the Light microscopy vulva (Plate 3). Gongylonema ingluvicola has The morphology of the nematodes was cuticular plates at the anterior end of their carried out by clearing in lacto-phenol with body (Plate 4) and the posterior part of an addition of a few drops of 10% lactic acid the male bears wide caudal alae and is and examined under the light microscope supported by numerous pedunculated at 10×, 20×, and 40× magnifi cations. The papillae (Plate 5). The female opens morphological structures were identifi ed posteriorly, showing the eggs and the vulva as according to the descriptions of Soulsby of this species (Plate 6). (1968). The anterior part of Ascaridia galli shows the presence of the cuticular plates

Pharynx

Lips

Plate 1. Lateral view of Cordon a male Acuaria spiralis showing the cordons at the anterior end

37 MALAYSIAN JOURNAL OF VETERINARY RESEARCH VOLUME 5 NO. 1 JANUARY 2014

Cordon Egg

Vulva

Anus

Plate 2. Anterior and posterior of female Plate 3. The vulva at the posterior part of Acuaria spiralis. the body of a female Acuaria spiralis.

Pharynx Esophagus

Lips

Cuticular plates

Plate 4. Anterior end, ventral view of male Plate 5. Posterior end of male Gongylonema Gongylonema ingluvicola showing the ingluvicola showing the caudal alae cuticular plates

Lips Vulva Cuticular plates

Egg

Plate 6. Vulva and eggs of Gongylonema Plate 7. Anterior part of female Ascaridia ingluvicola. galli showing cuticular plates

38 VOLUME 5 NO. 1 JANUARY 2014 MALAYSIAN JOURNAL OF VETERINARY RESEARCH

Anus Vulva

Egg

Plate 8. Posterior part of female Ascaridia Plate 9. Vulva and eggs of female Ascaridia galli showing the anus galli.

Spicules

Anus

Plate 10. Posterior end of male, Oxyspirura Plate 11. Posterior end of female Oxyspirura mansoni showing the two spicules of mansoni showing the anus different lengths

Lips

Egg Pharynx Vulva

Plate 12. Anterior end of Heterakis Plate 13. The vulva and eggs of Heterakis gallinarum, showing the bulb-structure at its gallinarum. alimentary tract

39 MALAYSIAN JOURNAL OF VETERINARY RESEARCH VOLUME 5 NO. 1 JANUARY 2014

only in the female (Plate 7). The vulva of covers the nematode body is a thin layer female Ascaridia galli is situated a short of hypodermis. distance anterior to the middle of the body In the present research, the anterior (Plate 8) with eggs in the uterus (Plate 9). end of Acuaria spiralis has four cuticular The left spicule of male Oxyspirura cordons beginning at sites of lips and mansoni is slender with the right long running to posterior end, not uniting and and stout at the posterior part (Plate 10). not forming marked curves. The cordons Female vulva of this species is situated in end before reaching the posterior end of the posterior part of the body (Plate 11). body (Skrjabin, 1969). Both sexes also have Heterakis gallinarum is characterised cordons; this species is easily separated by the presence of a bulb-like structure from others. The caudal end of male has at its alimentary tract (Plate 12), with the lateral alae and the posterior end of female presence of numerous eggs in the uterus is blunt. The vulva of this species lie in in the female (Plate 13). the posterior third of that body (Skrjabin, 1969). DISCUSSION Gongylonema ingluvicola is normally found in the upper digestive tracts of birds Bodies of nematodes are generally elongate and mammals. The cuticle of the anterior and cylindrical, tapering at both ends. A end is covered with large bosses (or certain degree of exists, cuticular plates), or irregular secutes and for the posterior end of males is commonly arranged in eight longitudinal rows (Gerald armed with special structures, for example & Larry, 1996). Gongylonema ingluvicola alae and papillae and are curved ventrally. has false lips and mouth capsule is present. Almost all males of nematodes are smaller Male left spicule is slander and have than females. Nematodes differ greatly in alate. Female vulva opens posteriorly and size, denpending on the species. Some of furthermore, lips of this species are small nematode species are microscopic and and also have a short pharynx with simple other species measure no more than 1 mm wall (Soulsby, 1968). (Cheng, 1964). Gerald and Larry (1996) stated According to Cheng (1964), the that the members of the Ascaridae are cuticle of parasitic nematodes is generally characterised by having three prominent smooth and the various structures such lips, valvulated bulb absent, and pre- as spines, bristles, warts, punctuations, cloacal sucker is sometimes present. papillae, striations and ridges may be It is large, thick, and yellowish-white present. The arrangements and positions with oral opening at the anterior end. of such structures are of taxonomic The mouth is surrounded by three large importance. Under the layer of cuticle that prominent lips and the esophagus is without a distinct posterior bulb. Male of

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this species is slightly smaller, and more and white in colour. Mouth surrounded slender than female. The tail of male has by three lips, a small buccal or pharynx. small caudal alae and bears a number of Esophagus ends in a well-developed bulb caudal papillae, most of which are short containing a valvular apparatus. The tail and thick. The posterior end is obliquely of male has large caudal alae extending truncated with pre-cloacal sucker oval or some distance down the sides of the body, circular on ventral side of tail. Spicules are bearing a number of caudal papillae and a well developed, protruding out at the anal prominent pre-cloacal sucker. The spicules opening. Female of this species is larger, are well developed, unequal, protruding stouter and vigorous than male. The tail out at anal opening. The tail of female tapers and the vulva is situated a short is also elongated, narrow and pointed. distance anterior to the middle of body. The vulva of the Heterakis gallinarum is The mouth of Oxyspirura mansoni situated at the middle of the body. is without defi ned lips and a short buccal capsule is usually present. The male is with REFERENCES or without caudal alae, preanal papillae simple, numerous and arranged in a linear 1. Anderson, R. (2000). Nematode Parasite of Vertebrates: Their Development and Transmission. Second Edition. row. The spicules of male are usually CABI Publishing, Wallingford, Oxon, UK. pp. 1-2, 23-25. unequal and the vulva in female may be 2. Blaxter, M.L., Page, A.P., Rudin, W., and Maizels, R.M. (1992). Nematode Surface Coats: Actively Evading either anterior or posterior. In male, the tail Immunity. Parasitol. Today 8: 243-47. is usually spiral, caudal alae absent and 3. Brown, H.W. and Neva. F.A. (1983). Basic Clinical Parasitology. Norwalk, Conn.: Appleton-Century-Crofts, the spicules unequal in length. In female, Inc. the vulva is located in front of the anus. 4. Butcher, G.D., Hogsette, J.A. and Jacobs, R.D. (1997). Nematode Parasites of Poultry (and where to fi nd them). The male tail is curved ventrally, without Science Department, Florida Cooperative caudal alae. The spicules are unequal, one Extension Service, Institute of Food and Agricultural Sciences, University of Florida. PS18. FL 32611. is slender and the other is long and stout. 5. Chan, Lai Keng and Siti Azizah Mohd. Nor. (2006). The vulva of this species is situated in the Teknik Mikroskopi dan Histologi. UniversitiSains Malaysia, Penang, Malaysia. pp. 90-124. posterior part of the body (Soulsby, 1968). 6. Campbell, Neil A and Reece, Jane B. (2005). . Soulsby (1968) stated that Heterakis Seventh Edition. Pearson Education, Inc., Benjamin Cummings, Sansome St., San Francisco. pp. 655-656. gallinarum has three well-defi ned lips, the 7. Cheng, Thomas C. (1964). The Biology of Animal esophagus with a short narrow anterior Parasites. W.B. Saunders Company, Philadelphia and London. pp. 390-472. portion (pharynx) and a long posterior 8. Chitwood, B.G. and Chitwood, M.B. (1937). An part ending in a bulb. The cuticle is usually Introduction to Nematology, sect. I, part I. Baltimore, Maryland: Monumental Printing Co. with lateral flanges. Esophagus has a 9. Chitwood, B.G. (1933). A revised classification of short narrow anterior portion (pharynx) nematode. J. Parasit., 20, 131 10. Chitwood, M.B. (1969). The systematic and biology of and with a long broader posterior portion some parasitic nematodes. Vol. III. New York, Academic ending in a well-developed bulb containing Press. pp. 223-244. a valvular apparatus. The worm is small

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