J Vet Clin 26(4) : 379-385 (2009) Light and Scanning Electron Microscopy of Tetragomphius procyonis (Ancylostomatoidea) from Wild Badger Hwa-Young Son, Yoon-Hee Oh, Hyeon-Cheol Kim* and Bae-Keun Park Institute of Veterinary Science and College of Veterinary Medicine, Chungnam National University, Daejeon 305-764, Korea *School of Veterinary Medicine, Kangwon National University, Chuncheon 201-100, Korea (Accepted : August 19, 2009) Abstract : Tetragomphius procyonis Baylis & Daubney, 1923 were obtained from the pancreatic duct of a naturally infected Eurasian badger, Meles meles, which was submitted to animal hospital for parasitic diagnosis from Gyeryongsan National Park in Korea. The hookworms were examined by light and scanning electron microscope. The length of body measured male 15.0-18.8 mm, female 21.5-25.5 mm, respectively. In both sex, the ventral cutting plates of oral margin are much reduced and elongated latero-dorsally, the dorsal cutting plate is located long follow doral margin of the oral opening. The buccal capsule is cup-shaped and thicken with four cusped tooth at its base. The copulatory bursa has elongated ventral lobes and their large rays are parallel, while the dorsal lobe with its supporting rays is slightly split in two. The slender spicules are filariform and very long (8.7-9.3 mm), and their tips are fused together. The hookworm has following characters: dorsal cone on the both sex, gubernaculum on the male and terminal spine on the female tail absent; vulva is opening in the juction of the fourths and fifths of the body; dorsal ray with two long stems. The eggs from the uterus are 16 cell stages and those from the rectal feces, 32 cell stages. The size of eggs is 77.48-83.45 × 50.75-63.38 µm. Key words : Tetragomphius procyonis, Eurasian badger, morphology. Introduction and badger differ from most hookworms. But the morphological study of those hookworms was carried out under the light The Ancylostomatoidea or hookworms are the parasites of microscopy. mammals, being most frequent in primates, carnivores and This paper reports morphological features of male and female ungulates, with a few species in other groups, including two of T. procyonis that were discovered from the pancreatic duct aquatic mammals (15). As with most zoological classifica- of wild badger in Korea under the light microscope and scan- tions there has been considerable discussion over specific nomen- ning electron microscope (SEM). clature and frequent changes have been proposed in specific names, arrangement and grouping of species (2,11,14,16,19). Materials and Methods The most recent review (13) proposed that the family Ancylo- stomatidae comprises two sub-families, Ancylostominae and Adult hookworms were harvested from naturally infected Bunostominae. The new classification of Ancylostomatidae Eurasian badger, Meles meles, transfered from Gyeryongsan proposed easily accommodates genera without cutting plates National Park. The worms were placed in lacto-phenol solu- or teeth (Globocephalus and Acheilostoma) or with small cutting tion (glycerin 20 ml, lactic acid 10 ml, phenol 10 g, D.W. 10 ml) plates (Tetragomphius and Bathmostomum). Also, Bunostom- for 24 hr to the transparency. The transparenced worms were inae is divided into tribes on the basis of morphological char- examined by light microscope. The parasites were previously acteristics, Acheilostominea and Bunostominea. The two washed with 0.1 M phosphate buffer pH 7.4 (PB) and fixed subgenera are placed in Acheilostominea. Acheilostoma is with 2.5% glutaraldehyde in PB at 4oC for 4 hrs. After wash- parasitic in large African thryonomid rodents, and Tetrago- ing with PB, the specimens were post-fixed with 1% osmum mphius is found in Asian badger and a raccoon. Genus Tet- tetroxide at 4oC for 4 hrs. Afterward, the specimens were ragomphius belong to subfamily Bunostominae and subgenera dehydrated in a graded ethyl alcohol series, dried by CO2 crit- Acheilostominea and has two species: T. procyonis (1) and T. ical point, coated with gold and examined by SEM. arctonycis (10). Those found in the pancreatic duct of raccoon The pancreas obtained from the badger was fixed in 10% neutral buffered formalin, and submitted to the histology service 1Corresponding author. for routine histological processing, paraffin-embedding, and E-mail : [email protected] H-E staining for the laboratory of the histopathological findings. 379 380 Hwa-Young Son, Yoon-Hee Oh, Hyeon-Cheol Kim and Bae-Keun Park Results Table 1. Comparison of Tetragomphius procyonis adult worm in relation to sex The total number of hookworm harvested from the main Male*(Mean) pancreatic duct was 91 worms of T. procyonis (male 35, female Body length 15.0-18.8 mm (16.7) 51, larvae 5) (Fig 1). But the larvae parasited in the branch of µ pancreatic duct were not counted. Histopathologically, the Body width 92.1-132.2 m (118.3) pancreatic duct was greatly dilated with severe periductal Distance of head to cervical papillae 123.2-191.4 µm (165.1) fibrosis but with minimal changes in the parenchyma of the Distance of head to esophageal end 194.3-221.8 µm (209.8) gland. The branches of pancreatic duct were infested with many Length of spicules 8.7-9.3 mm (9.1) of larvae (Fig 6). Female *(Mean) The sizes of adult hookworm were measured and summarized in Table 1; Male: The length of the body is 15.0-18.8 mm, the Body length 21.5-25.5 mm (23.7) maximum breadth is 92.1-132.2 µm (Fig 2). Excretory pore Body width 144.1-159.4 µm (151.25) and cervical papillae are situated 123.2-191.4 µm from anterior Distance of head to cervical papillae 128.9-231.8 µm (190.5) end. The esophagus is 194.3-221.8 µm. The spicules are 8.7- Distance of head to esophageal end 219.1-324.7 µm (248.3) 9.3 mm long (Figs 2, 3-3,5). Female: The length of the body Distance of vulva to tail end 1054.5-1440.8 µm (1288.6) µ is 21.5-25.5 mm, the maximum breadth is 144.1-159.4 m Distance of anus to tail end 37.6-66.68 µm (53.33) (Fig 2). Excretory pore and cervical papillae are situated 128.9- Size of eggs 77.48-83.45 × 50.75-63.83 µm 231.8 µm from anterior end. The esophagus is 219.1-324.7 µm *n=20 long. Distance of vulva to tail end is 1054.5-1440.8 µm. Dis- tance of anus to tail end is 37.6 - 66.68 µm. The eggs from the uterus are 16 cell stages (Fig 4-3) and those from the rectal feces, 32 cell stages (Fig 4-4). The size of eggs were mea- sured 77.48-83.45 × 50.75-63.38 µm. The features of cephalic region are generally the same in both males and female. The mouth opening is surrounded by 4 cephalic papillae and 2 amphidal pores (Figs 5-1,2,5). A pair of amphid is bilaterally identified by the presence of two lateral pores. The anterior end of worms is recurved dorsal direction (Fig 2), so that the well developed buccal capsule opens anterior-dorsally (Figs 3-1 and 5-1,2); it is relatively large and bears a pair of small cutting plates at its ventral margin and a small cutting plate, dorsally. The ventral plates of oral margin were much reduced and elongated latero-dor- sally. The dorsal cutting plate is located long follow dorsal margin of the oral opening. The buccal capsule is cup-shaped and thicken with four cusped tooth at its base (Figs 3-1 and 5-5,6). Those are a pair of three-cusped subventral teeth and a pair of roughly bifurcated subdorsal teeth. Those arise in the esophageal funnel and protrude into the buccal cavity. The dorsal cone does not found in the buccal capsule. A sim- ple excretory pore opens on the ventral portion between cer- vical papillae (Fig 5-8). The transverse cuticular striations of T. procyonis on the most of body surface are single, straight and deep, as shown in Fig 5-9. The cervical papillae are conical in shape and located bilaterally on the cuticule at the anterior-portion of esophageal end (Figs 3-2,5-8). Male: The copulatory bursa is well developed and shown in Figs 3-4,5,6 and 5-3,4. The striations on the surface of the Fig 1. T. procyonis (arrow) parasited in the pancreatic duct of bursa are not circular and are crossing. The bursa has elongated a badger. ventral lobes and their large rays are parallel, while the dorsal Fig 2. Light micrographs showing transparenced adult worms lobe with its supporting rays is slightly split in two. The same of T. procyonis. Note the long spicules (arrow). Vulva situated sized ventral rays run parallel and separated at the base of near the junction of the fourths and fifths (arrow). stem (Fig 3-4). Finger-shaped lateral rays are separated but Light and Scanning Electron Microscopy of Tetragomphius procyonis (Ancylostomatoidea) from Wild Badger 381 Fig 3. Light micrographs showing transparenced specimens of T. procyonis. 1. Lateral view of buccal capsule. The subventral (arrow head) and subdorsal teeth(arrow) arise in the esophageal funnel and protrude into the buccal cavity. 2. Cervical papilla (arrow) is located on the position of esophageal end bilaterally. E: esophagus. 3. Tail end of male. The very long spicles are slender and filaliform. 4. Male bursa and rays (lateral view). Finger-shaped lateral rays(□) are separated, but the medio- and postero-lateral rays are parallel.