Int. J. Biotechnol. and Allied Sci. 2 (3): 276-278 ISSN: 1597-9784 © Academic and Scientific Research Institute Publication, 2007 Available online at http://www.ijobas.org

DISTRIBUTION OF VARIOUS STAGES OF DEVELOPMENT IN PARAGONIMUS SPECIES IN

Nworie,1 O. Onyeagba2, R.A., Ugbogu2, O.C., Oduna1 T.J and Eze1, K 1. Applied Microbiology Department P. M. B. 053 Nigeria. 2. Microbiology Department University P.M.B. 2000 Uturu, Nigeria

Accepted August 2007 ABSTRACT: A total of 1922 freshwater snails (Oncomelania nosophora) (1000 from and 922 from ) were screened for presence of developmental stages of Paragonimus species using concentration technique. An overall prevalence rates of 456 (45.6%) and 215 (22.23%) were recorded for the different stages of the parasite at Ikwo and Afikpo respectively.Out of 1000 freshwater snails examined in Ikwo 105 (26.91%) showed the presence of cercariae, none revealed miracidia 92 (33.59%) sporocysts while rediae and embryonated eggs had 74 (22.65%) and 185 (28.53%) respectively. Freshwater snails examined at Afikpo North depicted that the 57 (43.20%) cercariae were infected sporocysts 51 (38.54%) rediae 47 (35.59%) and embryonated eggs 96 (34.92%) of the Paragonimus were harboured by the snails. However miracidia stage was not found in the snails examined. The high level of Paragonimus developmental stages in snails examined cells for prompt education on the best way to control paragonimiasis.

Key words: Paragonimus, Oncomelania nosophora , miracidium, sporocyst, redia and cercaria Corresponding Author: Nworie, O.

INTRODUCTION Paragonimiasis is a zoonotic infection with a lung-dwelling flukes of the genus paragonima. These flukes have a complex life cycle involving snails, crabs and man (Defrain and Hooker, 2004). More than fifty species of paragonimus have been reported from different countries of the world but only nine are known to infect humans. The genus, paragonimus shows greatest diversity in relation to geographical distribution host etc. In Korea, the predominant species is P.westermani (Choi,1990) but in Africa especially in Nigeria, P.africanus and P.uterobilateralis are most reoccurring species (Asor et al., 2003) The developmental stages of this parasite in the freshwater snails are: egg, miracidium, sporocyst, redia and cercaria. The eggs are golden brown in colour, oval in shape and are provided with a flattened operculum. The size of the egg is between 80 to 120µm. Within the snail the egg is transformed into a ciliated miracidium which later develop into sporocyst. Sporocyst is a bag-like structure with embryonic germ cells; also the sporocyst in turn produces redia which is an elongated and non ciliated larva. The redia continues growing within the snail giving rise to several individuals of the tadpole-like stage called cercaria. The cercaria bears a tail which helps it in movement inside water but the tail is lost when developing into an infective stage, metacercaria which is only found in crustaceans and humans (Sachs and Voeiker,1982). This work was carried out to curb the role of snails in the transmission of paragonimiasis in Ebonyi State Nigeria.

MATERIALS AND METHODS Study area: The study was carried out in Ebonyi State South Eastern Nigeria. The state has a population of about 1.8 million and a total land area of about 5935km2 which gives a population density of over 300 persons per sq km. Ebonyi State is located between latitudes 7030' east and 8030' west and longitudes 5040', north and 6045', south within the rain forest zone of Nigeria. The fertile and rich soils of Ebonyi State encourage large scale agriculture. The area under study is noted for high snail availability and consumption. Sample size: A total of two thousand nine hundred and twenty two fresh water snails were collected and examined. Sample collection Live and healthy snails were bought and some collected from the riverine base Ikwo and Afikpo North LGAs of Ebonyi State. The collection was by scooping using net baskets.

276 Int. J. Biotechnol. and Allied Sci. Nworie et al.

Then the fresh water snails were placed in a small container filled with pond water. They were picked randomly in few numbers with sample bucket. The samples were carried to the laboratory for analysis. Methodology The methods of Davis et al., (1994) and Cheesbrough (2004) were employed in this work. The fresh water snails (Oncomelania nosophora) were examined using concentration technique. They were dissected using a pair of scissors and dissecting forceps after breaking the shell with a small hammer. The tissues of the snails were grinded using a mortar and pestle and were centrifuged at 2000 rpm for 10mins; the sediments were then examined using 40X objectives.

RESULTS AND DISCUSSION Snails (Oncomelina nosophora) are one of the sources of first class proteins. They are highly cherished by the people of Ebonyi State especially the Abakiliki zone. Unfortunately , some developmental stages of the fluke, Paragonimus are harboured by snails which could later be transmitted to crustaceans (crayfish and crabs) which man will consume thus resulting in Paragonimus infection. In this study, a total of 185(28.53%), 92(33.59%), 74(22.65%) and 105(26.91) embryonated eggs, sporocysts, rediae and cercariae respectively were observed in one thousand Oncomelina nosophora examined in Ikwo (Table 1). However these findings do not agree with the findings of Asor et al.,(2003) that reported 6.1% infection rate for Afropomus balanoides in the Cross River basin. At Afikpo, 96(34.92%) embryonated eggs, 51(38.54%) sporocysts, 47(35.59%) rediae and 57(43.20%) cercariae were known to have infected the nine hundred and twenty two snails tested in this area (Table II). The sporocyst and cercariae seem to be the most occurring developmental stages in the snails examined in both zones. The miracidia stage was not observed in the screened snails in both Ikwo and Afikpo zones. These snails pose a risk to humans who consume them as they are potential sources of Paragonimus infection. Earlier studies have implicated snails as intermediate hosts of Paragonimus (Amunarriz, 1991; Aka et al., 1991). Metacercarial infection of Paragonimus westermani has been reported in markets in Seoul (Cho et al., 1991; Choi, 1990). Paragonomiasis has been reported in various parts of Eastern and South eastern Nigeria (Nwokolo, 1972; Arene et al., 1998; Asor et al., 2003). The infection rate in this study varies from location to location, the highest infection rates were observed in Ekpa-Omaka in Ikwo LGA while Uawara had the highest infection rate in Afikpo North LGA. The farming practices of these localities expose them to eating these snails which serve as intermediate host for Paragonimus infection. There is need to under study the affected population in these areas and to increase the level of awareness of the potential of infection in this area.

Table 1. Distribution of the developmental stages of Paragonimus in snails harvested from Ikwo L.G.A. Location Number Number infected percentage infection (%) Examined EE M S R C EE M S R C Ekpa-Omaka 238 47 0 19 28 31 19.74 0 7.98 1.18 1.30 Ekpa – Elu 90 16 0 17 13 14 1.78 0 1.89 1 .44 1.56 Alike 186 34 0 11 07 17 1.83 0 5.91 3.76 9.14 Ameka 210 47 0 16 09 24 2.24 0 7.62 4.29 1.14 Okpoitumo 130 16 0 12 04 09 1.23 0 9.23 3.08 6.92 Ndufu Echara 146 25 0 17 13 10 1.71 0 1.16 8.90 6.85 Total 1000 185 0 92 74 105 28.53 0 33.59 22.65 26.91

Key: EE : Embryonated egg, M: Miracidium, S : Sporocyst, C : Cercaria

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Table 2. Distribution of the developmental stages of paragonimus in snails harvested from Afikpo North L.G.A. Location Number Examined Number infected Percentage infected (% ) EE M S R C EE M S R C Itim 132 16 0 09 10 11 1.21 0 6.82 7.58 8.33 Oziza 132 12 0 06 07 09 9.09 0 4.55 5.30 6.82 Anohia 132 21 0 13 10 11 1.59 0 9.85 7.58 8.33 Ugwuegu 137 19 0 11 08 09 1.37 0 8.03 5.84 6.57 Ohaisu 130 06 0 02 03 05 4.62 0 1.54 2.31 3.85 Mkpoghoro 130 02 0 00 00 01 1.54 0 0.00 0.00 0.77 Uawara 129 20 0 10 09 11 15.50 0 7.75 6.98 8.53 Total 922 96 0 51 47 57 34.92 0 38.54 35.59 43.20 Key: EE: Embryonated egg, M : Miracidium, S : Sporocyst, R : Redia, C : Cercaria

REFERENCES Aka, N. A.,Allabi, A. C., Dretfuss, G., Kinde, G. D., Tawo, L.R., Bouteille, B., Avode, G., Anagonous, S. Y., Ginafon, M., Masso0ugbodiji, A., Dumas, M (1991 ). Observations on the first case of human Paragonomiasis and potential intermediate hosts of Paragonimus species. Bull Soc. Pathology Ext, 92: 191 – 194. Amunarriz, M. (1991 ). Intermediate hosts of Paragonimus in the Eastern Amazomic regionof Ecuador. Trop. Med. Parasitology, 42: 164 – 166. Arene, F. O., Ibanga, E. and Asor, J.E. (1998 ). Epidemiology of paragonimiasis in Cross River Basin, Nigeria: Pargonimus uterobilateralis in Yakurr Local Government Area. Public Health, 112: 119 – 122 Asor, J.E., Ibanga, E.S and Arene, F.O.I (2003). The Epidemiology of Pulmonary Paragonomiasis in Cross River Basin Nigeria: Update on infection prevalence and distribution of the snail and crab intermediate hosts. Mary Slessor Journal of Medicine 3:5-12 Cheesbrough, M. ( 2002 ). District laboratory practice in tropical countries. Part 1, Cambridge University press, London. Pp. 178- 306 Choi, D. (1990 ) Paragonimus and Paragonmiasis in Korea. Kisaengehuung – hak Chapehi, 1: 79- 102. Cho, S.Y., Kang, S.V., Kong, Y. and Yong, H. J. (1991). Metacercarial infection of Paragonimus westermani in markets in Seoul. Korean J. Parasitol 29: 189 – 192. Davis, G.M, Chen, C.E., Kang, Z.B. AND Livi, V.Y. ( 1994 ). Snail hosts of paragonimus in Asia and America. Biomed. Environ – Sci, 7: 369 – 382. Defrain, M. and Hooker, R. (2005 ). North American Paragonimiasis: case report of serve clinical infection – selected reports Journals of pathology. Pathological society of Great Britain and Ireland, 121: 1368 – 1372. Nwokolo, C ( 1972 ). Out break of Paragonimiasis. In Eastern Nigeria. J Trop. Med. Hyg. 1:32 – 33. Sachs, R and Voeiker, J . ( 1982 ). Human paragonimiasis caused by paragonimus uterobilaterlis in Liberia and Guinea, West Africa. Tropen med parasitology, 33: 15 – 16.

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