Biodiversity of nematode composition of millipede trigoniulus corallius (Diplopoda: Trigoniulidae)

Journal of Science – 2015, Vol. 4 (4), 166 – 175<br /> <br /> Part D: Natural Sciences, Technology and Environment<br /> <br /> BIODIVERSITY OF NEMATODE COMPOSITION OF MILLIPEDE-TRIGONIULUS<br /> CORALLIUS (DIPLOPODA: TRIGONIULIDAE)<br /> Bui Thi Dung1, Pham Van Luc2, Hoang Van Hien1, Nguyen Van Duc3<br /> 1<br /> <br /> M.S., Vietnam Academy of Science and Technology, Vietnam<br /> Asoc. Prof. Dr., Vietnam Academy of Science and Technology, Vietnam<br /> 3<br /> PhD., Vietnam Academy of Science and Technology, Vietnam<br /> 2<br /> <br /> Information:<br /> Received: 09/07/2015<br /> Accepted: 05/08/2015<br /> Published: 07/09/2015<br /> <br /> ABSTRACT<br /> <br /> Millipede Trignoiulus corallines (Gervais, 1847) were collected in two national<br /> parks (Tam Dao and Cuc Phuong). The Skrjabin dissection method was used to<br /> investigate parasitic nematode in millipedes. The results showed the high<br /> prevalence of nematode infection in millipeded (83,33%; 30/36; intensity 1-137<br /> Keywords:<br /> nematode per millipede). Ten nematode species were identified (Rhigonema<br /> Nematode, millipede,<br /> critesi, Rhigonema nesoboli, Cattiena fansipanis, Heth bifidispiculum, Heth<br /> Trignoiulus corallines, Tam Dao,<br /> vietnamensis, Thelastoma collare, Thelastoma meadsi, Thelastoma medecassa,<br /> Cuc Phuong<br /> Rhabditis sp., Ichthyocephaloides sumbatus). Among them, two new species<br /> (Heth vietnamensis và Cattiena fansipanis) were previously described in<br /> Vietnam. The other species were described in other countries but are now first<br /> describes in Vietnamese specimens.<br /> <br /> In Vietnam, millipede Trignoiulus corallinus<br /> (Gervais, 1847) is common. However, there is a<br /> lack of information about parasites in millipede.<br /> Therefore, the present study on biodiversity of<br /> nematode species is necessary in order to assess<br /> the biodiversity of parasites in millipede and the<br /> species with potential for use as biological<br /> control.<br /> <br /> 1. INTRODUCTION<br /> Millipede (Diplopoda) plays an important role in<br /> the ecosystem. Millipede is involved in the<br /> decomposition of plants. They feed on leaves,<br /> rotten wood, fungi and plants with soft roots, and<br /> green moss (Hopkin & Read, 1992). Millipede is<br /> generally not harmful to humans and animals.<br /> They usually live in moist areas, so they are<br /> especially important for tropical forests.<br /> <br /> 2. MATERIALS AND METHODS<br /> <br /> Although millipede is a beneficial animal group<br /> they will become pests when developed with<br /> greater density. Millipede can destroy crops such<br /> as potatoes, tomatoes, and watermelon. There are<br /> many studies showing that millipede are harmful<br /> organisms and must be controlled. In the control<br /> measures, biological control method using<br /> parasitic nematodes to reduce millipede density is<br /> used (McKillup, van Harter, & Neves, 1991).<br /> <br /> Millipede (Trignoiulus corallines) was collected<br /> by hand in Tam Dao (Vinh Phuc province) and<br /> Cuc Phuong (Ninh Binh province) national parks<br /> (Fig. 1). Millipede was transferred to the<br /> laboratory for surgery.<br /> The surgical method: dissect from the anus to the<br /> head, and then the pointed needles attached at two<br /> sides. Intestines were moved to the saline solution<br /> 0.9% and examined under stereomicroscope to<br /> 166<br /> <br /> Journal of Science – 2015, Vol. 4 (4), 166 – 175<br /> <br /> Part D: Natural Sciences, Technology and Environment<br /> <br /> collect large nematode. After that, the continuous<br /> filtering method was used to detect small<br /> nematodes in intestinal fluid.<br /> <br /> the permanent slides were made follow the<br /> Seinhort method (Seinhort, 1959).<br /> Nematode were measured, photographed and<br /> drawn under Olympus-CH40 microscope.<br /> Nematode was identified using the Skrjabin key<br /> (Skrjabin, 1991).<br /> <br /> Nematode fixation: the collected nematode was<br /> fixed in 4% formaline solution. The nematode<br /> temporary slides were made with glycerine and<br /> <br /> Figure 1. Millipede - Trigoniulus corallinus (Gervais, 1847)<br /> <br /> 2. Species: R. nesoboli Garcia & Morffe, 2014<br /> <br /> 3. RESULTS AND DISCUSSION<br /> <br /> Family Carnoyidae Filipjeve, 1934<br /> <br /> 3.1 Prevalence of nematode infection in<br /> millipede in two national parks<br /> <br /> Genus Cattiena Hunt & Spiridonov, 2001<br /> <br /> Total thirty of 36 examined millipedes were<br /> infected by nematode, prevalence 83.33%. The<br /> prevalence of nematode infection in millipede was<br /> similar in Tam Dao and Cuc Phuong National<br /> Parks;<br /> 85.0%<br /> (17/20,<br /> intensity<br /> 1-137<br /> nematode/host), 81.3% (13/16, intensity 1-81<br /> nematode/host) respectively.<br /> <br /> 3. Species Cattiena fansipanis Malysheva &<br /> Luc, 2012<br /> Family Hethidae Skrjabin & Shikhobalova, 1951<br /> Genus Heth Cobb, 1898<br /> 4. Species: Heth bifidispiculum Adamson, 1983<br /> 5. Species: Heth vietnamensis Malysheva &<br /> Spiridonov, 2010<br /> <br /> 3.2 Nematode composition in millipede<br /> <br /> Family Thelastomatidae Travassos, 1929<br /> <br /> Ten nematode species belonging to 6 genus, 6<br /> families, 3 orders were identified. The<br /> classification below:<br /> <br /> Genus Thelastoma Leidy, 1849<br /> 6. Species: Thelastoma collare Upton, 1983<br /> 7. Species: Th. meadsi Clark, 1978<br /> 8. Species: Th. medecassa Waerebeke, 1967<br /> <br /> Order Rhabditida Chitwood, 1933<br /> Family Rigonematidae Chitwood, 1935<br /> Genus Rigonema Cobb, 1898<br /> <br /> Family Rhabditidae Öley, 1880<br /> <br /> 1. Species: Rhigonema critesi Ramirez, 1974<br /> <br /> Genus Rhabditis Dujadin, 1845<br /> 167<br /> <br /> Journal of Science – 2015, Vol. 4 (4), 166 – 175<br /> <br /> Part D: Natural Sciences, Technology and Environment<br /> <br /> Male. Body length 3.5-4.26 mm, body maximum<br /> width 0.256-0.28 mm. Oesophagus length 0.2720.292 mm, maximum at posterior end 0.1-0.12<br /> mm. Nerver ring to the anterior end 0.16-0.168<br /> mm. Two equal spicules or slightly longer<br /> different, simple structure and do not kintin,<br /> spicule length 0.288-0.32 mm. Gubernaculum is<br /> absent. Tail curved. Tail has 5-9 papilae.<br /> <br /> 9. Species Rhabditis sp.<br /> Order Spirurida Chitwood, 1933<br /> Family Ichthyocephalidae Travassos & Kloss,<br /> 1958<br /> Genus Ichtyocephaloides Artigas, 1926<br /> 10. Species: Ichthyocephaloides sumbatus<br /> Malysheva & Spiridonov, 2013<br /> <br /> Female. Body length 3.5-4.26 mm; body<br /> maximum width 0.256-0.28 mm. Oesophagus<br /> length 0.272-0.29 mm. Corpus measurement 0.10.116 x 0.12-0.14 mm. Nerve ring and expretory<br /> pore to anterior end 0.128-0.192 mm, 0.28-0.328<br /> mm, respectively. Vulva at the posterior end,<br /> vulva to the anterior end 2.6-3.44 mm. Tail is<br /> short and cone-shaped, length 0.148-0.16 mm.<br /> Body width at the anus 0.108-0.128 mm. Uterus<br /> extends posteriorly. Ovary is long and contains<br /> full eggs. Eggs have round-shaped, thick wall and<br /> measurement 0.074-0.084 x 0.06-0.066 mm.<br /> <br /> Morphological description of parasitic nematode<br /> in millipede<br /> 1. Rhigonema critesi Ramírez, 1974 (Figure<br /> 2)<br /> Body is thick and long. Cuticun has translucent<br /> striations. Bucal cavity is surrounded by cuticun<br /> ring with three lips and four papillae. Oesophagus<br /> is short and without isthmus. Corpus is round.<br /> Excretory pore opens ventrally at corpus-bulb<br /> junction.<br /> <br /> Figure 2. Rhigonema critesi Ramirez, 1974<br /> (a, male; b, female; c, head; d, vulva; e, male’s tail)<br /> <br /> 168<br /> <br /> Journal of Science – 2015, Vol. 4 (4), 166 – 175<br /> <br /> Part D: Natural Sciences, Technology and Environment<br /> <br /> 2. Rhigonema nesoboli García & Morffe,<br /> 2014 (Figure 3, only female)<br /> <br /> 0.24 mm. Corpus measurement 0.05x0.08 mm.<br /> Excretory pore opens ventrally at corpus-bulb<br /> junction. Vulva at posterior part of body, vulve to<br /> posterior end 1.15-1.23 mm. Tail sharp and length<br /> 0.35-0.46 mm.<br /> <br /> Female. Body length 2.6-3.5 mm. Buccal cavity is<br /> surrounded by cutin ring with three lips.<br /> Oesophagus is short, without isthmus. Corpus is<br /> round, without valve. Oesophagus length 0.21-<br /> <br /> Figure 3. Rhigonema nesoboli Garcia & Morffe, 2014<br /> (a, female’s head; b, vulva; c, tail)<br /> <br /> valve. One spicule with simple structure, spicule<br /> length 0.13-0.135 mm. Guberculatum located near<br /> spicule, length 0.73-0.77 mm.<br /> <br /> 3. Cattiena fansipanis Malysheva & Luc,<br /> 2012 (Fig. 4)<br /> Small body with clear striations. There are four<br /> cephalic papillae, cephalic has spinal ring. Large<br /> bucal cavity. Long tail with needle-shaped, and<br /> lateral curved.<br /> <br /> Female. Body length 3.2-3.9 mm. Nerve ring to<br /> anterior end 0.13-0.15 mm. Excretory pore opens<br /> at the middle of oesophagus. Oesophagus length<br /> 0.24-0.3 mm. Corpus has valve. Vulva at posterior<br /> end. Tail is sharp and needle-shaped, tail length<br /> 0.85 mm.<br /> <br /> Male: body length 1.63-2.15 mm. Nerve ring to<br /> the anterior end 0.18 mm. Excretory pore opens at<br /> the anterior part, at middle of oesophagus.<br /> Oesophagus length 0.26-0.33 mm. Corpus has<br /> <br /> 169<br /> <br /> Journal of Science – 2015, Vol. 4 (4), 166 – 175<br /> <br /> Part D: Natural Sciences, Technology and Environment<br /> <br /> Figure 4. Cattiena fansipanis Malysheva & Luc, 2012<br /> (a, male; b, male’s head; c, male’s tail; d, female; e, female’s head; f, female’s tail)<br /> <br /> length 0.35-0.42 mm, Corpus measurement 0.710.80 mm. Vulva open near the anus, vulva to<br /> posterior end 0.55-0.65 mm. Tail is long and<br /> sharp.<br /> <br /> 4. Heth bifidispiculum Adamson, 1983<br /> (Figure 5, only female)<br /> Female. Typical of species is prominent head that<br /> established cutin plate. Buccal cavity has four<br /> papilae. Body length 1.9-2.3 mm. Oeshophagus<br /> <br /> Figure 5. Heth bifidispiculum Adamson, 1983<br /> (a, female; b, female’s head; c, female’s tail)<br /> <br /> 170<br /> <br />