Molluscicidal Assessment of Aqueous Extract of Moringa Oleifera Lam Seed on Bulinus Snail for the Control of Schistosomiasis

South Asian Journal of Research in Microbiology

7(1): 30-39, 2020; Article no.SAJRM.58268 ISSN: 2582-1989

Molluscicidal Assessment of Aqueous Extract of Moringa oleifera Lam Seed on Bulinus Snail for the Control of Schistosomiasis

Umetiti Chukwuemeka Nnamdi1, Ene Paschal Chuka2*, Orji Nkeiru Mary Ann1, Umedum Chinelo U.3, Ugwu Kenneth Chukwudi1 and Ikeanumba Michael Okwudiri4

1Department of Biological Sciences, Chukwuemeka Odumegwu Ojukwu University, Anambra State, Nigeria. 2Department of Med. Lab. Tech, School of Health Technology, Oji River, Enugu State, Nigeria. 3Department of Microbiology, Chukwuemeka Odumegwu Ojukwu University, Anambra State, Nigeria. 4Department of Biology, Alvan Ikoku College of Education, Owerri, Imo State, Nigeria.

Authors’ contributions

This work was carried out in collaboration among all authors. Author UCN Designed the proposal, procured the materials and the reagent used for the study, involved in collection, processing of the samples, data entry and analysis. Author EPC Helped in collection and processing of the samples, wrote the manuscript and submitted it for publication. Author ONMA Supervised the study and reviewed the manuscript. Author UCU co-supervised the study and reviewed the manuscript. Author UKC helped in collection and processing of the samples. Author IMO helped in data analysis and result interpretation. All authors read and approved the final manuscript.

Article Information

DOI: 10.9734/SAJRM/2020/ v7i130162 Editor(s): (1) Dr. Luciana Furlaneto-Maia, Technological University of Parana, Brazil. Reviewers: (1) Anandaramajayan Nallathambi, Sri Lakshmi Narayana Institute of Medical Sciences, India. (2) Kiran, India. Complete Peer review History: http://www.sdiarticle4.com/review-history/58268

Received 10 April 2020 Original Research Article Accepted 16 June 2020 Published 01 July 2020

ABSTRACT Aims: This research is essentially on the Molluscicidal assessment of Aqueous extract of Moringa oleifera Lam seed on Bulinus Snail for the control of Schistosomiasis. Study Design: This is a controlled study where a total of 810 Bulinus snails were collected from three different streams with each stream representing a community from each of the three senatorial zones of Anambra state where schistosomiasis was endemic. Aqueous extract of Moringa oleifera (Lam) seed at different concentrations were used on the Bulinus snails to determine their molluscidal properties. ______

*Corresponding author: Email: [email protected]; Umetiti et al.; SAJRM, 7(1): 30-39, 2020; Article no.SAJRM.58268

Place and Duration of Study: Two hundred and Seventy (270) Bulinus snails were collected from streams representing each of the three senatorial zones of Anambra state viz; Obutu Lake, Omogho town, Orumba North Local Government Area (Anambra South), Agulu Lake, Agulu town, Aniocha Local Government Area (Anambra Central) and Omambra River, Omor town, Ayamelum Local Government Area (Anambra North) This research was carried out between November 2018 to April 2019. Methodology: Aqueous dilutions of the grinded Moringa oleifera (Lam) seed were exposed to the Bulinus snails from the various streams for 24 hours, after which the snails were removed from the experimental test solution and washed thoroughly with dechlorinated tap water and transferred to containers with fresh dechlorinated tap water for another 24 hours of recovery. The snails were incubated at 28 ± 5°C and fed with lettuce leaves. Results: Aqueous Moringa oleifera Lam seed extract was toxic to Bulinus adult snails in a dose dependent manner and the total Lethal Concentration (LC50 and LC90) values determined after 24 hours exposure from the whole streams were 468 ppm and 813 ppm respectively. Conclusion: Aqueous seed extract of Moringa oleifera Lam was toxic to Bulinus snail even at low concentrations.

Keywords: Bulinus; Moringa oleifera; schistosomiasis; LC50; LC90; streams; Anambra State.

ABBREVIATIONS Bulinus is a genus of small tropical fresh water snails, function as intermediate hosts for the LC50 : 50% Lethal Concentration schistosomiasis blood fluke [7]. LC90 : 90% Lethal Concentration NTDs : Neglected Tropical Diseases Control of Bulinus spp. comprises an important cMoL : Coagulant Moringa oleifera Lam element in integrated strategies for reducing the lectin spread of schistosomiasis, and the single WsMoL : Water-soluble Moringa oleifera Lam molluscicide recommended by the World Health lectin Organization is the organic compound PPM : Parts Per Million niclosamide. This molluscicide possesses low toxicity to mammals; however, it is toxic to fishes, 1. INTRODUCTION amphibians, and crustaceans [8,9]. Also due to the high cost, environmental contamination, and Schistosomiasis is a neglected tropical parasitic possible development of snail resistance to infection [1,2], with high prevalence in Nigeria [3]. chemical molluscicides, natural molluscicides are It is a snail-borne trematode infection, a blood rapidly being developed [10]. Many plant extracts flukes from the genus Schistosoma which are potential molluscicides that are according to [4], is endemic in 74-76 environmentally friendly, less toxic and are less developing countries especially in impoverished likely to cause snails to develop resistance [11]. communities without access to a sound public health system and has been ranked second to Moringa oleifera Lam plants have provided a malaria in terms of socio-economic and public number of useful clinical agents that prove to health significance [5,6]. have considerable potentials as source of new Snail intermediate hosts play active roles in the drugs [12]. The aqueous extract of Moringa transmission of Schistosomiasis infections in oleifera Lam seeds contain bioactive molecules Africa. They are the sites of an intense including saponins, lectins, [13], and volatile oils multiplication of parasites. Most of the diseases [14]. There are two types of lectins: coagulant caused by snail-borne trematodes are prevalent Moringa oleifera Lam lectin (cMoL) which is a in the tropic and sub-tropic regions of the world, basic protein and water-soluble; Moringa oleifera and the medical and economic burden of these lectin (WSMoL) which is an acidic protein [15, diseases are often neglected which is why they 16]. These proteins have ability to agglutinate are called Neglected Tropical Diseases (NTDs). erythrocytes [17] and show coagulant activity that reduces water turbidity [18]. Recent studies A lot of controls strategies have been employed showed that M. oleifera Lam lectins had ovicidal in combating this disease, the snail control effects on Aedes aegypti [19] and insecticidal strategies are considered a priority for the potential against Anagasta kuehniella (especially reduction of schistosomiasis transmission. cMoL) [20]. Also, the seed powder of Moringa

31 Umetiti et al.; SAJRM, 7(1): 30-39, 2020; Article no.SAJRM.58268 oleifera Lam had a molluscicidal activity against snail collection was made possible with the the snails Biomphalaria glabrata and Physa assistance of a wooden canoe. The scoop net marmorata [21,22]. Therefore, the use of was placed under the aquatic plants and medicinal plants which grow abundantly in areas intermittently checked for trapped snails. This where Schistosomiasis is endemic may become was repeatedly done until the numbers of snails a useful complement either as molluscicides or needed were obtained. chemotherapeutic for the control of the disease. Medicinal plants are promising choices that may 2.2.3 Criteria for snail selection grow the scope of molluscicides accessible for controlling of Bulinus snails as these plants are The snails must be from the above streams less costly, safer and having a high level of representing the three senatorial zones in degradability [23]. Anambra State, the snail must be motile, Only adult snails between 10 – 13 mm were collected, 2. METHODOLOGY embryos and small sized snails were left out. Only those snails free from any infection and 2.1 Study Area devoid of any strange colors were used.

The study area Anambra state is made up of 2.2.4 Procession of snails three senatorial zones viz: North, Central and South. The senatorial districts were used as Adult Bulinus snail which is a fresh water snails yardstick for partitioning of the study area. were used for this study. This was carried out in Anambra state is in the south eastern part of in a semi controlled environment with the Nigeria and lies on the latitude 5°40′ N and average daily temperature range of 28°C to 33°C 6°45N and 6°35′E and 6°30′E). Hydrologically, in open plastic tanks measuring 50 × 23 × 17 cm Anambra composition include rivers, streams, containing filtered de-chlorinated fresh water. lakes and ponds with River Niger and Anambra River being the largest and longest with several The snails were fed daily on dry previously boiled tributaries that give rise to many streams and lettuce. The snails from the various streams were streamlets. Thus the study was conducted in reared in separate tanks in the laboratory. Water three selected communities from the three in bowls was changed twice a week to eliminate L.G.A’s. The towns were Agulu (6.0923°N, faecal droppings and fouling. The snails were 7.0411°E), Omogho 6.3642°N, 6.8163°E) and kept for a period of two weeks to acclimatize to Omor (6.5224°N, 6.9281°E) Fig. 1. Other rivers laboratory conditions before use in the and streams include; Nkissi, Anakwaeze, Ulasi, experiment. Otolo, Ekulo, Aghammiri, Mamu, Obizi, Egi, Oyi, Ariba, Aroka. After 24 hours challenge with the aqueous Moringa oleifera Lam, the dead snails were 2.2 Study Design counted and the still viable snails were recovered 2.2.1 Sources of bulinus snail and introduced into fresh water for two weeks to check for after effects of the aqueous Moringa A total of 810 Bulinus snails were used for the oleifera Lam exposure. Within the 2 weeks dead study. Two hundred and Seventy (270) of them as well as unhealthy, algal infected snails were from streams representing each of the three removed as soon as discovered. senatorial zones of Anambra state viz; Obutu Lake, Omogho town, Orumba North Local 2.2.5 Sources and preparation of the Moringa Government Area (Anambra South), Agulu oleifera lam seeds Lake, Agulu town, Aniocha Local Government Area (Anambra Central) and Omambra River, The seeds used in the study were collected from Omor town, Ayamelum Local Government Area a 4 year’s old Moringa oleifera plant. The (Anambra North). Moringa oleifera seeds were collected washed and air dried. The seed-lobes were grinded into 2.2.2 Collection of snails powdery form. 10 g of powdery seed were weighed and added into 1000 mL of The snails were collected from the streams/rivers dechlorinated water. It was shaken vigorously in each of the senatorial zones with the help of and allowed to stand for 3 hours. The mixture the scoop net. The snail usually attaches itself was filtered and the filtrate collected and oven beneath aquatic plants like water lettuce. This dried for 24 hours. It was filtered to get the solid

32 Umetiti et al.; SAJRM, 7(1): 30-39, 2020; Article no.SAJRM.58268 concentrate. The seed powder was weighed and of Aqueous Moringa oleifera Lam seed in added directly to 1000 mL of filtered water in Omogho town, Orumba North Local Government individual beakers at eight different Area of Anambra south Senatorial Zone. The concentrations: various logarithm of concentrations were obtained, likewise the Probit Unit values for the 2.2.6 Molluscidal bioassay average death per each concentration were also The Bulinus spp adulticidal bioassay was obtained. The graph in Fig. 1 shows the performed according to the methods of the logarithm of the different concentrations plotted [24] following the recommendation of the Expert against the different probit units. These were Committee on the Bilharzia. used to compute the LC50 and LC90 values. After the computation, the LC50 and LC90 for aqueous In each assay, ten healthy adult snails with Moringa oleifera Lam seed extract in Omogho uniform size (shell diameter of 10 –13 mm) were Senatorial zone were calculated to be 467.7 ppm immersed for 24 hours in a glass jar containing and 812.8 ppm respectively. 200 ml of water. The treatments were: seed powder (0.2, 0.4, 0.6, 0.8, 1.0, 1.5 and 2.0 g/l, −1 Fig. 2 like its counterpart in Anambra south copper (II) carbonate (50 μg mL ; positive Senatorial Zone shows the results obtained from control) and just filtered tap water (negative Agulu town in Aniocha Local Government Area of control). After the exposure period of 24 hours, Anambra Central Senatorial Zone. The fatality of the snails were removed from the experimental aqueous Moringa oleifera seeds was seen to test solution and washed thoroughly with de- progressively increase as the concentration of chlorinated tap water and transferred to the aqueous Moringa oleifera seeds increased. containers with fresh de-chlorinated tap water for The two apex concentrations of 1500 and 2000 another 24 hours of recovery. The snails were ppm recorded 100% fatalities, while the least incubated at 28 ± 5°C and fed with lettuce concentration of 200 ppm had an average of leaves. 3.3% Bulinus spp. deaths. This signifies the Evaluation of snail survival was made after 14 toxicity of aqueous Moringa oleifera seeds even days (2 weeks) and survival rates (%) were at that low concentration. obtained for each treatment. Snails were considered dead if they could not move nor The various logarithms of concentrations were retract out of the shell, with their bodies and also obtained, so were the Probit Unit values for shells discolored [24]. the average death per each concentration. The graph also shows the logarithm of the different And then, the percentages of observed concentrations plotted against the different probit mortalities were recorded. Bioassays were units to compute the LC50 and LC90 values performed in triplicate for each concentration. obtained from the zone. After the computation, the LC50 and LC90 for aqueous Moringa oleifera 2.2.7 Calculation of Lethal Concentration (LC) Lam seed extract in Omogho Senatorial zone This was done using a combination of Microsoft were calculated to be 436.5 ppm and 794.3 ppm Excel, Probit table, Data Analysis and respectively. Regression. The scenario in Omor town, Ayamelum L.G.A, 2.3 Statistical Analysis Anambra North Senatorial Zone was what is seen in Fig. 3 where the various logarithm of The data collected were analyzed using SPSS concentrations and the Probit Unit values for the version 17. For the bioassay, Probit analysis by average death per each concentration were [25] was used to calculate the lethal obtained and used to calculate the LC50 and Concentration (LC). Data analysis was also done LC90 from the zone (Fig. 3). After the using Regression. Significant differences computation, the LC50 and LC90 for aqueous between treatments groups used in the Moringa oleifera Lam seed extract in Omogho bioassays were analyzed using Student’s t-test Senatorial zone were calculated to be 426.6 ppm (significance at P < 0.05). and 812.8 respectively.

3. RESULTS Fig. 4 is the collation of Percentage death of Bulinus spp treated with different concentrations Fig. 1 presents the average Percentage death of of aqueous Moringa oleifera Lam seed in all the Bulinus spp treated with different concentrations Senatorial Zones. Also, the various logarithm of

33 Umetiti et al.; SAJRM, 7(1): 30-39, 2020; Article no.SAJRM.58268

10 8 6 4 ProbitUnit 2 0 2.3 2.6 2.8 2.9 3 3.2 3.2 Logarithm of Concentration y = 5.27x + (-9.07)…

Fig. 1. LC50 and LC90 of Bulinus spp treated with different concentrations of aqueous Moringa oleifera seed in the triple tests done in Omogho town To compute LC50 To compute LC90 y = 5.27x + (-9.07) y = 5.27x + (-8.6) Where y = 5.0, x = 2.67 Where y = 6.28, x=2.91 Antilog 2.67 = 467.7 Antilog 2.91=812.8 LC50 = 468 ppm LC90 = 813 ppm

10 8 6 4 ProbitUnit 2 0 2.3 2.6 2.8 2.9 3 3.2 3.2 Logarithm of Concentration y = 4.8x + (-7.67)…

Fig. 2. LC50 and LC90 of Bulinus spp treated with different concentrations of aqueous Moringa oleifera seed in the triple tests done in Agulu town To compute LC50 To compute LC90 y = 4.8x + (-7.67) y = 4.8x + (-7.67) Where y = 5.0, x = 2.64 Where y = 6.28, x=2.90 Antilog 2.64 = 436.5 Antilog 2.90=794.3 LC50 = 436,5 ppm LC90 = 794.3 ppm concentrations and the Probit Unit values for the Anambra North, Central and South were 427, average death per each concentration were 437 and 468 ppm respectively, while obtained and used to calculate the LC50 and LC90 the LC90 were 813, 794 and 813 ppm from the zone. Like the calculations from the respectively. various zones the graph in Fig. 4 was used to calculate the LC50 and LC90 which were Fig. 6 shows a graph where the total mortality calculated to be 468 ppm and 813 ppm rate (%) of the Bulinus snail was plotted against respectively. the various concentration (PPM) of aqueous Moringa oleifera Lam seed. The responses of Fig. 5 shows the bar chart representation of LC50 snails tested in terms of probit death were found and LC90 of the various towns from the various to form a linear relationship between percentage senatorial zones at a glance. The LC50 for concentration (ppm) and probit unit of the

34 Umetiti et al.; SAJRM, 7(1): 30-39, 2020; Article no.SAJRM.58268 extracts. Aqueous Moringa oleifera seeds was Lam seed increases, the mortality rate of the toxic to Bulinus adults in a dose-dependent Bulinus snails also increases until a certain manner. What this means is that as the threshold was reached where all the Bulinus concentration of the aqueous Moringa oleifera snails were ultimately destroyed.

9 8 7 6 5 4

Pobit Unit Pobit 3 2 1 0 2.3 2.6 2.8 2.9 3 3.2 3.2 Logarithm of Concentration y = 4.65x + (-7.27)

Fig. 3. LC50 and LC90 of Bulinus spp treated with different concentrations of aqueous Moringa oleifera seed in the triple tests done in Omor town To compute LC50 To compute LC90 y = 4.65x + (-7.27) y = 4.65x + (-7.27) Where y = 5.0, x = 2.63 Where y = 6.28, x=2.91 Antilog 2.63 = 426.6 Antilog 2.91=812.8 LC50 = 426.6 ppm LC90 = 812.8 ppm

9 8 7 6 5 4

ProbitUnit 3 2 1 0 2.3 2.6 2.8 2.9 3 3.2 3.2 Logarithm of Concentration y = 5.34x + (-9.28)

Fig. 4. LC50 and LC90 of Bulinus spp treated with different concentrations of aqueous Moringa oleifera seed in the average triple tests done in all the zones To compute LC50 To compute LC90 y = 5.34x + (-9.28) y = 5.34x + (-9.28) Where y = 5.0, x = 2.67 Where y = 6.28, x=2.91 Antilog 2.63 = 467.7 Antilog 2.91=812.8 LC50 = 468 ppm LC90 = 813 ppm

35 Umetiti et al.; SAJRM, 7(1): 30-39, 2020; Article no.SAJRM.58268

1000 800 600 LC50 400 LC90 200 0 North Central South

Fig. 5. LC 50 and LC90 of all the zones at a glance

120 100 80 60 40 Mortality (%)Mortality 20 0 200 400 600 800 1000 1500 2000

Concentration (ppm)

Fig. 6. Mortality rates (%) of Bulinus at different concentrations of the aqueous Moringa oleifera seeds

Table 1 shows the survival rate of Bulinus snails scourge of schistosomiasis in these senatorial exposed to sub-lethal concentrations of M. zones worked on, which comprised all the oleifera Lam seeds (<600 ppm) after exposure senatorial zones that made up Anambra state. followed by 2 weeks of recovery. In this table there was a uniform progressive decrease in The aqueous Moringa oleifera Lam seeds extract survival rate as the sub-lethal concentration was toxic to Bulinus adult snails in a dose- increases. Most of the control recorded 100% dependent manner. This deduction was made Bulinus snail survival followed by concentration based on the calculated LC50 and LC90 in all the of 200 ppm where the Bulinus snail survival rate three senatorial zones studied. For every was greater than that of 400 ppm which in turn senatorial zone the experiment was done in was greater than 600 ppm. This pattern showed triplicate, and the average probit dead recorded. the same uniformity in all the zones. Also the As represented from Figs 1 – 4; LC 50 and 90 difference in the various concentrations were were calculated using a complex data analysis significantly greater to the next one preceding it involving regression and probit analysis. There (P<0.05), e.g, the survived Bulinus snails in the was the same uniform toxicity pattern in all the controls were significantly greater than the three senatorial zones studied in that the survived Bulinus snails in the 200 ppm group aqueous Moringa oleifera Lam seeds extract which in turn was significantly greater than the indicated low toxicity at ppm of 200, and as from snails in 400 ppm group etc. 1000 ppm the toxicity affected more than 90% percent of the snails. Concentrations greater 4. DISCUSSION than 1000 ppm were very fatal for all the Bulinus snails. The least concentration of 200 ppm in all The study revealed that the aqueous Moringa the pooled Bulinus spp treated with Moringa oleifera Lam seeds extract have the potential of oleifera seeds aqueous extract averaged 4% controlling Bulinus snails and by extension, the mortality, with a progressive increase in the

36 Umetiti et al.; SAJRM, 7(1): 30-39, 2020; Article no.SAJRM.58268 fatalities of the snails as the concentration Comparable perceptions were recorded by [26] increases. The LC50 and LC90 from the different who stated that exposure of B. alexandrina snails zones vary, though not significantly (P>0.05).The to methanol extracts of Euphorbia splendens, difference in this variation could not be Atriplex stylosa, and Guayacum officinalis led to ascertained in this work since there was no death a significant decrease in their survival and growth in the negative control but it might be due to the rates. The reduction in survival rate was due to individual differences in their various ecosystem the snails overcoming the destructive impact of and habitat. When pooled together, the values these toxic compounds through discharging it to determined from the three senatorial zones after the surrounding media or by biodegrading it to 24-h exposure were 468 ppm and 813 ppm for non-poisonous by-products [27]. LC and LC respectively. 50 90 This also correlated with the work of [28] whose results demonstrated that the grinded seed of Fig. 6 showed that the responses of snails Moringa oleifera Lam was lethal for B. glabrata tested, in terms of probit death, were found to (LC50 0.419 g/L and LC 1.021 g/l) and P. form a linear relationship between percentage 90 marmorata (LC 0.339 g/L and LC 0.789 g/L) concentration (ppm) and probit unit of the 50 90 but had no effect against M. tuberculatus . They extracts. This means that as the concentration of concluded that the seed powder of Moringa the aqueous Moringa oleifera seed increased, oleifera, never tested before with this purpose, the mortality rate of the Bulinus snails also has molluscicidal activity, being lethal to the increased until a certain threshold was reached snails B. glabrata and P. marmorata and non- where all the Bulinus snails were ultimately lethal to M. tuberculatus. Also [29] found that the destroyed. LC50 and LC90 values for Moringa oleifera leaves aqueous extract on Bulinus truncatus were 392 The above results agree with findings of [21], ppm and 595 ppm respectively. Also The LC who confirmed that the seed powder of Moringa 50 and LC90 values for Moringa oleifera leaves oleifera had a molluscicidal activity against the aqueous extract on cercariae of Schistosoma snails Biomphalaria glabrata and Physamar heamatobium were found to be 378 ppm and 618 morata and stated that the snails retracted into ppm respectively. Their study showed that the shell and suffered hemorrhage after treatment. effect of Moringa oleifera leaves on cercariae was more than its effect on B. truncates. At the Table 1 shows that survival rates of adult Bulinus highest and least concentration of M. oleifera snails were markedly reduced post their leaves (750 – 250) respectively, the death rate exposure to sub-lethal concentrations of the for the snails was found to be 100% and 10% aqueous seed extract. The main aim of this was respectively, compared with 98.6% and 20% for to see whether the snails will recover after cercariae at the same concentration. They exposure to sub-lethal concentrations (<600 concluded that the Balanites aegyptiaca ppm) by recovering and feeding them using fresh mesocarp aqueous extract and Moringa oleifera water and boiled lettuce. From the table, two leaves have the potential of controlling Bulinus snails from the negative control died for no truncatus and cerceriae of Schistosoma apparent reason from Omogho town, but the rest heamatobium. from other towns were intact. There was a decrease in snail survival after 2 weeks as the Studies also showed that Moringa oleifera lectins sub-lethal concentration increased which in had ovicidal effects on Aedes aegypti [19] and epidemiological term signifies that some snails insecticidal potential against Anagasta kuehniella though alive after addition of the extract, will later [20]. Also, the seed powder of Moringa oleifera succumb to the deleterious effects of the Moringa had a molluscicidal activity against the snails oleifera seed extract. On the other hand, some Biomphalaria glabrata and Physa marmorata will shake off the deleterious effect and thrive. [21,22].

Table 1. Survival rate (%) of Bulinus snails exposed to sub-lethal concentrations of M. oleifera lam seeds after exposure followed by 2 weeks of recovery

Concentration (ppm) Control 200 400 600 Omogho 98 93.4 89.1 53.5 Agulu 100 95.1 74 50.1 Ayamelum 100 83.8 69.1 43.7

37 Umetiti et al.; SAJRM, 7(1): 30-39, 2020; Article no.SAJRM.58268

These results agree with that of [30] who stated cercariacidal effects of Glinus lotoides that water extract of Moringa oleifera Lam seeds fruits. Res Notes. 2014;7:1- 22. had lethal action against Aedes aegypti larvae 2. Rizk MZ, Aly HF. Recent therapeutic and eggs. Also, [31] studied the ovicidal action of approaches in control of parasitic diseases Moringa oleifera flower extract and stated that it with special reference to schistosomiasis. had delayed the development of treated embryos Int J Adv Res. 2015;1:957–971. and reasoned that the flower extract caused 3. Ekwunife CA, Ukaga CN, Okafor FC. changes in the physiology of snails, which Urinary schistosomiasis in Anambra State, interfered with the production of eggs. Nigeria. Nig J Parasitolo. 2005;25:127-131. 4. World Health Organization. The control of It can be concluded that the seed powder of Schistosomiasis. World Health Moringa oleifera Lam never tested before with Organization Technical Report Series. this purpose, has molluscicidal activity, being 2017;830:34 – 39. lethal to Bulinus snails tested, even at low 5. Ekpo F, Laja-Deile A, Oluwole S, et al. concentration. Urinary schistosomiasis among pre-school children in rural community near Abeokuta, 5. CONCLUSIONS Nigeria. Para & Vect. 2010;3:58-68. 6. Nwoke BEB, Dozie INS, Nwoke JC, Aqueous seed extract of Moringa oleifera Lam is Amosike JC. Human schistosomiasis and toxic to Bulinus snail even at low concentration. Nigerian environment and climate change. Due to its non-toxic, nutritional and water purifier Bio-Res. 2004;2(1):103-114. adjuvant characteristics The characteristics of 7. Kane RA, Stothard JR Emery AM, et al. this plant are a starter for a new line of research Molecular characterization of freshwater for more specific products in the control of snails in the genus Bulinus: A role for aquatic snails, being safer and with low barcodes? Para & Vect. 2008;1:15. environmental impact which is associated to the 8. Salem HK, Omram NE, Eissam SH, et al. protection of biodiversity of other non-target Induction of teratogenesis of freshwater organism. snail (Biomphalaria alexandrina) using the molluscicide niclosamide. Afr J Sci. CONSENT AND ETHICAL APPROVAL 2014;2:255-268. 9. World Health Organization. The control of A letter was obtained from the Head of schistosomiasis. Report of a WHO Expert Department of Biological Sciences Committee. World Health Organization Chukwuemeka Odumegwu Ojukwu University to Technical Report Series. 2014;728:65-67. Anambra State Ministry of Education. Informed 10. Yang F, Long E, Wen J, et al. Linalool, consents were obtained from the various derived from Cinnamomum camphora leaf community leaders through a letter from the state extracts, possesses molluscicidal activity Ministry of Education to assist in the collection of against Oncomelania hupensis and inhibits materials/samples for this research work. infection of Schistosoma japonicum. Para ACKNOWLEDGEMENTS & Vect. 2014;7:40-47. 11. Rawani A, Ghosh A, Chandra G. The Authors acknowledge the support of the Laboratory evaluation of molluscicidal & various community leaders and Chief Priests of mosquito larvicidal activities of leaves the various rivers/lakes who consented for this of Solanum nigrum. Ind J. Med Res. research using their rivers/lakes and also to the 2014;140:285–295. various fishermen for allowing the use of their 12. Choy SY, Prasad KM, Wu TY. Utilization of canoes and aiding in the collection of the snails. plant-based natural coagulants as future alternatives towards sustainable water COMPETING INTERESTS clarification. J Env Sci. 2014;26:2178– 2189. Authors have declared that no competing 13. Araújo LC, Aguiar JS, Napoleão TH. interests exist. Evaluation of cytotoxic and anti- inflammatory activities of extracts and REFERENCES lectins from Moringa oleifera seeds. PLoS One. 2013;8:81973. 1. Kiros G, Erko B, Mekonnen Y. Laboratory 14. Kayode RM, Afolayan AJ. Cytotoxicity and assessment of molluscicidal and effect of extraction methods on the

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