032321027 Ezenwaka

Endo-Helminth Parasites of Anurans of Ahoada Main Town and Joinkrama, Ahoada, Rivers State, Nigeria

Ezenwaka, C.O.1; *Amuzie, C.C.2

1Department of Biology, Faculty of Science, Federal University Otuoke, Bayelsa State, Nigeria. 2Parasitology and Entomology Laboratory, Department of Animal and Environmental Biology, Rivers State University, Port Harcourt, Nigeria.

ARTICLE INFO ABSTRACT

Article No.: 032321027 Research into the species of anurans and their associated helminth parasites is essential for conservation and scientific purposes. Two reports on the helminth Type: Research parasites of anurans from Ahoada area of Rivers State, Nigeria, have been documented. This research, therefore examines anurans from unexplored locations in Ahoada (namely, Ahoada Main Town and Joinkrama). Search and capture techniques were employed in sampling and standard parasitological methods were Accepted: 23/03/2021 used. Helminth parasites were fixed in appropriate fixatives. Both anuran and Published: 25/03/2021 parasitic species were identified using applicable keys. Prevalence and mean intensity of infection were computed following conventional methods. Twenty-eight *Corresponding Author anuran species comprised of Hoplobatrachus occipitalis, Ptychadena bibroni, P. Amuzie, C.C. oxyrhynchus, P. pumilio, P. mascareniensis and Hymenochirus sp. were collected from Ahoada Main Town. These were infected with fourteen helminth parasite E-mail: nmaamuzie@ gmail. com species (namely monogeneans [Polystoma aeschlimanni and P. pricei], digeneans [Mesocoelium monodi, Diplodiscus fischthalicus, Ganeo africana and Keywords: Helminths; Metahaematoloechus exoterorchis], cestodes [Cylindrotaenia jaegerskioeldii], and Amphibians; Ahoada; Joinkrama; nematodes [encysted ascaridida larvae, Rhabdias sp., Oswaldocruzia hoepplii, Niger Delta Cosmocerca ornata, Chabaudus leberrei and Amplicaecum africanum]. At Joinkrama, thirty amphibian specimens comprised of toads (Sclerophrys maculata) and frogs (Hylarana galamensis, Hoplobatrachus occipitalis, Ptychadena bibroni and Ptychadena mascareniensis) were examined. Ten helminth species (namely, acanthocephalan cystacanths, cestodes (Cylindrotaenia jaegerskioeldii), trematodes (Diplodiscus fischthalicus, Mesocoelium monodi, Metahaematoloechus exoterorchis) and nematodes (Amplicaecum africanum, Cosmocerca ornata, Rhabdias sp., Rhabdias africanus and Oswaldocruzia hoepplii) were isolated from infected hosts. Prevalence of infection ranged from 6.25% to 100% at both locations; mean intensity ranged from one to twenty-one parasites per infected hosts at Ahoada Main Town and one to thirteen at Joinkrama. The altered nature of the habitats surveyed is thought to have influenced both the anuran and endohelminth parasites diversity recorded.

Greener Journal of Biological Sciences, vol. 11, no. 1, pp. 14-20, 2021 Ezenwaka and Amuzie / Greener Journal of Biological Sciences 15

INTRODUCTION Mesocoelium monodi and Mesocoelium sp.), one cestode (Baerietta jaegerskioeldii), acanthocephalan Anurans include frogs and toads and comprise of about cystacanths, one pentastomid (Raillietiella sp.) and three thirty families (Flament et al., 2011). Research into the monogeneans (Polystoma aeschlimanni, P. baeri and P. helminth parasites of anurans in Rivers State, Nigeria, pricei). has progressed significantly. Akani et al. (2011) Among the documented reports on helminth pioneered the research examining anuran species from parasites of anurans from Rivers State, Nigeria, we some residential areas in Port Harcourt, Rivers State, found information on anurans from the Ahoada area of Nigeria. Subsequently, other researchers including the State in Aisien et al. (2017b) and Amuzie et al. Amuzie et al. (2016), Aisien et al. (2017a), Amuzie (2019). This present research at Ahoada Main Town and (2017), Amuzie et al. (2019) and Okere et al. (2019) Joinkrama represent efforts to fully document the conducted studies in several locations of the State. species of anurans and their helminth parasites across The anuran species examined by these Rivers State and the entire Niger Delta region. researchers included the bufonids (Sclerophrys spp.), ptychadenids (Ptychadena spp.), hyperolids (Afrixalus MATERIALS AND METHODS spp. and Hyperolius spp.), pipids (Silurana tropicalis, Hymenochirus sp.), Ranids (Hylarana galamensis) and Sample Locations, Methods and Duration dicroglossids (Hoplobatrachus occipitalis) among others. The endo-helminth parasites of these host species were Anuran samples were taken from two locations: Ahoada also documented and a common observation among Main Town in Ahoada East Local Government Area and these reports is the greater diversity of nematode Joinkrama in Ahoada West Local Government Area, parasites above other helminth classes. For instance, Rivers State, Nigeria. Samples were collected from both Amuzie et al. (2019) reported eight nematode species locations using the search and capture technique from (Amplicaecum africanum, ascaridia larvae, temporary pools of water on grass fields (Plate 1). This Batracocamallanus siluranae, Chabaudus leberrei, was done between the hours of 8:00 to 10:00pm during Cosmocerca ornata, Oswaldocruzia hoepplii, Rhabdias each expedition. Sampling was conducted from May to africanus and two other Rhabdias spp.), six trematodes July, 2018, at Ahaoada Main Town, and in September, (Prosotocus exovitellosus, Ganeo africana, Diplodiscus 2020, at Joinkrama. Anurans were identified after fischthalicus, Metahaematoloechus exoterorchis, Roedel (2000).

Plate 1: Temporary pools of water sampled for anurans at Joinkrama, Ahoada West LGA, Rivers State, Nigeria

Laboratory Examination Laboratory, Department of Animal and Environmental Biology, Rivers State University, Port Harcourt, for Anuran samples were transported in wet and aerated examination. They were anesthetized and measured to containers to the Entomology and Parasitology determine the wet body weight (using a top-local 15

16 Ezenwaka and Amuzie / Greener Journal of Biological Sciences electronic balance) and snout-vent length (using a metre pumilio, P. mascareniensis and Hymenochirus sp. rule). Fourteen helminth parasite species were isolated from They were thereafter dissected for the isolation of the infected hosts. The parasites were monogeneans helminth parasites. The lungs, gastrointestinal tract, (Polystoma aeschlimanni and P. pricei), digeneans urinary bladder and body cavity were examined in Petri (Mesocoelium monodi, Diplodiscus fischthalicus, Ganeo dishes half-filled with 0.9% normal saline solution. africana and Metahaematoloechus exoterorchis), Parasites were picked up using pipettes and fixed cestodes (Cylindrotaenia jaegerskioeldii), and appropriately in well labeled bottles: nematodes were nematodes (encysted ascaridida larvae (Plate 2), stretched in hot water and fixed in 70% alcohol; Rhabdias sp., Oswaldocruzia hoepplii, Cosmocerca monogeneans, trematodes and cestodes were flattened ornata, Chabaudus leberrei and Amplicaecum in 5%formol saline under cover slip pressure and fixed in africanum). the same solution. The overall prevalence of infection with parasites ranged from 6.25% to 100% (Table 1). Among the Identification of Helminth Endoparasites and monogeneans, Polystoma aeschlimanni infected Computation of Prevalence and Mean Intensity of Ptychadena pumilio at a prevalence of 20% while P. Infection pricei infected Ptychadena mascareniensis at a prevalence of 12.5%; mean intensity of infection was two The helminth endoparasites were identified with the help per infected host for both parasites. The digeneans, of identification keys (Prudhoe and Bray, 1982; Khalil et Mesocoelium monodi and Metahaematoloechus al., 1994) using compound microscopes. The number of exoterorchis infected Ptychadena mascareniensis and helminthes recovered was recorded per predilection site. Hoplobatrachus occipitalis, respectively, at a prevalence Prevalence and mean intensity of infection were of 37.50%. Diplodiscus fischthalicus and Ganeo africana computed following the formula of Bush et al. (1997). both infected H. occipitalis at 6.3% prevalence. The only cestode, Cylindrotaenia jaegerskioeldii, was isolated from P. mascarenienesis at 25% RESULTS AND DISCUSSION prevalence and mean intensity of about five parasites per infected host. Among the nematodes, the highest Twenty-eight anurans were collected from Ahoada Main prevalence of 100% was reached by Ascaridia larva-2 in Town- Ahoada East- over the sampling period and Ptychadena oxyrhynchus, and Amplicaecum africanum included the following species: Hoplobatrachus in Ptychadena bibroni. occipitalis, Ptychadena bibroni, P. oxyrhynchus, P.

Table 1: Overall Prevalence (P%) and mean intensity of helminth infection in amphibians of Ahoada Main Town, Ahoada East L.G.A., Rivers State, Nigeria Parasite Host Predilection Site Prevalence (%) Mean intensity (±sem) Monogenea Polystoma P. pumilio Urinary bladder 20.00 2.0±0.00 aeschlimanni P. pricei P. mascareniensis Urinary bladder 12.50 2.0±0.00

Digenea M. monodi P. mascareniensis Small intestine 37.5 12.0±5.3 D. fischthalicus H. occipitalis Rectum 6.25 1.00±0.0 G. africana H. occipitalis Small intestine 6.25 2.00±0.0 M. exoterorchis H. occipitalis Lungs 37.5 12.5±2.2

Cestoda C. jaegerskioeldii P. mascareniensis Small intestine 25.0 4.5±2.5

Nematoda Ascarid larva 1 P. mascareniensis Body cavity 12.5 2.00±0.0 Ascarid larva 2 H. occipitalis Body cavity 56.25 6.4±1.8 P. oxyrhynchus Body cavity 100.0 7.00+0.0 Rhabdias sp. P. mascareniensis Lungs 12.5 21.0±0.0 O. hoepplii P. mascareniensis Small intestine 25.0 2.5 ±0.5 A. africanum P. mascareniensis Small intestine 12.5 1.00±0.0 P. bribroni Small intestine 100.0 4.00±0.0 H. occipitalis Small intestine 6.25 1.00±0.0 C. ornata P. mascareniensis Rectum 37.5 10.3±7.3 C. leberrei H. occipitalis Small intestine 6.25 2.00±0.0 ±standard deviation Ezenwaka and Amuzie / Greener Journal of Biological Sciences 17

At Joinkrama – Ahoada West LGA- a total of thirty in H. occipitalis. Whereas prevalence was 88% in H. amphibian specimens were captured and examined for occipitalis, it was 33% in S. maculata. Mean intensity of helminth parasites. These were comprised of toads infection was 13.5 parasites per infected host in H. (Sclerophrys maculata) and frogs (Hylarana galamensis, occipitalis, and 3.0 parasites per infected host in S. Hoplobatrachus occipitalis, Ptychadena bibroni and maculata. Ptychadena mascareniensis). A total of ten helminth Another trematode, M. monodi, infected H. species, including members of the Acanthocephala, galamensis, H. occipitalis and S. maculata, at Cestoda, Trematoda and Nematoda, were recovered prevalence rates of 20%, 22% and 66%, respectively. from the anuran hosts. The parasites were namely, Mean intensity of infection was higher in H. galamensis acanthocephalan cystacanths (Acanthocephala), (11 parasites per infected host), followed by H. Cylindrotaenia jaegerskioeldii (Cestoda), Diplodiscus occipitalis (5.5 parasites per infected host) and fischthalicus, Mesocoelium monodi, Sclerophrys maculata (1.5 parasites per infected host). Metahaematoloechus exoterorchis (Trematoda), Five nematode species were recovered from the Amplicaecum africanum, Cosmocerca ornata, Rhabdias specimens examined in this project. Amplicaecum sp., Rhabdias africanus and Oswaldocruzia hoepplii africanum was found only in Sclerophrys maculata; (Nematoda). prevalence and mean intensity of infection were 33% The acanthocephalan cystacanths were isolated and 3.0, respectively. Cosmocerca ornata was isolated from the body cavity of Ptychadena mascareniensis. from four of the five host species examined. These were Prevalence of infection was 100.0% and the mean H. galamensis, H. occipitalis, P. bibroni and S. maculata, intensity of infection was seven parasites per infected and both prevalence and mean intensity of infection host. The tapeworm, C. jagerskioeldii, infected only P. were highest in S. maculata. bibroni at a prevalence of 50.0% and mean intensity of Two species of Rhabdias were isolated, Rhabdias one parasite per infected host. africanus from S. maculata and an unidentified species Among the trematodes, Diplodiscus fischthalicus from Hylarana galamensis. Oswaldocruzia hoepplii was infected only Hoplobatrachus occipitalis at a prevalence isolated from Ptychadena bibroni at a prevalence of of 11% and mean intensity of one parasite per infected 100% and mean intensity of infection was 1.5 parasites host. Metahaematolechus exoterorchis infected both H. per infected host (Table 2). occipitalis and Sclerophrys maculata. However, both prevalence and mean intensity of infection were higher

18 Ezenwaka and Amuzie / Greener Journal of Biological Sciences

Table 2: Prevalence (%) and mean intensity (MI) of parasite infection in amphibian hosts of Joinkrama, Ahoada-West L.G.A., Rivers State, Nigeria Parasite Host(s) Predilection site Prevalence (%) MI ±Stdev

Acanthocephala Acanthocephalan Ptychadena mascareniensis Body cavity 100.0 7.0±0.0 cystacanths

Cestoda Cylindrotaenia Ptychadena bibroni Small intestine 50.0 1.0±0.0 jaegerskioeldii

Trematoda Diplodiscus Hoplobatrachus occipitalis Rectum 11.11 1.0±0.0 fischthalicus Metahaematoloechus H. occipitalis Lungs 88.89 13.5±17.4 exoterorchis Sclerophrys maculata Lungs 33.33 3.0±0.0 Mesocoelium monodi Hylarana galamensis Small intestine 20.0 11.33±12.1 H. occipitalis Small intestine 22.22 5.5±2.1 S. maculata Small intestine 66.67 1.5±0.7

Nematoda Amplicaecum africanum S. maculata Small intestine 33.33 3.0±0.0 Cosmocerca ornata H. galamensis Rectum 20.0 3.0±2.0 H. occipitalis Rectum 11.11 1.0±0.0 S. maculata Rectum 100.0 7.6±1.5 P. bibroni Rectum 50.0 1.0±0.0 Rhabdias sp. H. galamensis Lungs 53.33 8.1±9.0 Rhabdias africanus S. maculatus Lungs 33.33 3.0±0.0 Oswaldocruzia hoepplii P. bibroni Small intestine 100.0 1.5±0.7 Stdev: standard deviation

A B

Plate 2: Ascaridida larva- A, Type 1; B, Type 2- isolated from Anurans, Ahaoada Main Town, Ahoada East LGA, Rivers State, Nigeria

The anuran composition at both locations were similar Hylarana galamensis were found only at Joinkrama. This being comprised of species such as Hoplobatracchus disparity is thought to be due to the short duration of occipitalis, Ptychadena bibroni and Ptychadena sampling at both locations because similar altered mascareniensis. Ptychadena oxyrhynchus, P. pumilio habitats have been reported to support all of these and Hymenochirus sp. were only encountered at species (Amuzie and Akani, 2017; Amuzie et al., 2019). Ahoada Main Town, whereas Sclerophrys maculata and Habitat structure affects anuran diversity as a result of its Ezenwaka and Amuzie / Greener Journal of Biological Sciences 19 effect on the availability of preferred niches of varying cestodes, digeneans and nematodes. The altered nature species of anurans (Pearman, 1997). The undisturbed of the habitats surveyed is thought to have influenced rainforest would be expected to hold a higher diversity of both the anuran and endohelminth parasites diversity amphibian species than anthropogenically disturbed recorded. landscapes (Ernst et al., 2007). Anthropogenic alterations also affect parasite dynamics enhancing or reducing prevalence as the case REFERENCES may be. For instance, Roznik et al. (2015) reported that removal of rainforest canopy cover reduced the Akani, G.C., Luiselli, L., Amuzie, C.C. and Wokem, G.N. prevalence of the fungus Batrachochytrium (2011). Helminth community structure of three dendrobatidis in the rainforest frogs, Litoria rheocola. afrotropical anuran species: A test of the interactive- The parasites reported in the anuran samples examined versus-isolationist parasite communities hypothesis. in the present research reflect the parasite fauna of Web Ecology, 11, 11-19. 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Cite this Article: Ezenwaka, CO; Amuzie, CC (2021). Endo-Helminth Parasites of Anurans of Ahoada Main Town and Joinkrama, Ahoada, Rivers State, Nigeria. Greener Journal of Biological Sciences, 11(1): 14-20.