Larvicidal Activity of Phyllanthus fraternus Powder in Suppressing Dermestes maculatus Degeer (Coleoptera: Dermestidae) Infestation on Smoked African Catfish (Clarias gariepinus)  

J. M. Adesina1 , A. R. Jose2 , O. O. Adetuyi3 , D. A. Olorunfemi4
1. Department of Crop, Soil and Pest Management Technology, Rufus Giwa Polytechnic, P. M. B. 1019, Owo, Ondo State, Nigeria
2. Department of Science Laboratory Technology, Rufus Giwa Polytechnic, P. M. B. 1019, Owo, Ondo State, Nigeria
3. Department of Fisheries and Aquaculture Technology, Rufus Giwa Polytechnic, P. M. B. 1019, Owo, Ondo State, Nigeria
4. Department of Agricultural Technology, Rufus Giwa Polytechnic, P. M. B. 1019, Owo, Ondo State, Nigeria
Author    Correspondence author
International Journal of Aquaculture, 2014, Vol. 4, No. 11   doi: 10.5376/ija.2014.04.0011
Received: 07 Feb., 2014    Accepted: 12 Mar., 2014    Published: 12 Apr., 2014
© 2014 BioPublisher Publishing Platform
This is an open access article published under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
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Adesina et al., 2014, Larvicidal Activity of Phyllanthus fraternus Powder in Suppressing Dermestes maculatus Degeer (Coleoptera: Dermestidae) Infestation on Smoked African Catfish (Clarias gariepinus), International Journal of Aquaculture, Vol.4, No.11: 67-72 (doi: 10.5376/ija.2014.04.0011)


The efficacy of the plant powder of Phyllanthus fraternus was evaluated under tropical storage condition against larvae of the leather beetle Dermestes maculatus.The experiment was in two sets, set A which are in concentration 0g, 1.0g, 2.0g, 2.5g, 3.0g/15g of smoked fish and in B 0g, 3.5g, 4.0g, 4.5g, 5.0g powder admixed with 20g of smoked fish; while all the treatment containing 0g were without plant powder which served as control, all the treatment were in triplicates. Newly emerged (0 - 72 hours old) larvae of D. maculatus was introduced. Evaluation of the potency of the plant powder was based on the larvae mortality and the adult emergence.  The weight losses in fish muscle in the treated and untreated samples were compared as index of fish damage during storage. The result showed that higher plant powder concentration were significantly (P<0.05) effective in killing larvae stage of the insect while at 1st and 2nd weeks after infestation (WAI) non-significant (P>0.05) was observed except for 1st WAI in set A treatment. Adult emergence was significantly (P<0.05) inhibited in treated fish in set A and weight loss due to insect infestation was greatly suppressed by the higher dosage rate of the plant powder compared to control. The phytochemical analysis of P. fraternus powder revealed the presence of Tannins, Saponins, Alkaloids and Kelakellani (glycosides), this might be responsible for the larvicidal activity of P. fraternus powder against larval of D. maculatus. This finding revealed that P. fraternus powder toxicity and efficacy is dosage dependent and suggested that the plant could be used by poor resource fish farmers, processors and marketers in protecting smoked-dried fish against D. maculatus during processing, transportation, marketing and storage.

Phyllanthus fraternus; Adult emergence; Efficacy; Larvae mortality; Toxicity; Phytochemical; Potency

Fish constitutes about 50% of total animal protein intake in Nigeria. Since the cost of beef increases daily and owing to its nutritional and health benefit of fish, its wide acceptability on the menu table of most Nigerians irrespective of socio-economic status, age and religious background could not be over emphasised.
Fish protein has a high biological value because it is composed of a very high profile of essential amino acids, notably lysine, methionine and isoleucine, which are relatively deficient in animal protein (Abolagba et al., 2011). It is a good source of thiamine, riboflavin, vitamins A, B, D, E and K and respectable amount of minerals such as phosphorus, calcium and iron, iodine, fluorine and magnesium (Yem et al., 2006); it is equally high in polysaturated fatty acids which are important factor for lowering blood cholesterol level (Conquer and Holub, 2002; Okunade, 2011).
The artisanal sector which contributes about 90 per cent of the total fish production in the country depends mainly on smoking and drying as the means of preserving their catch that cannot be sold fresh, probably because the process requires a minimum of capital investments.
Smoke-dried fish is susceptible to attack by insect especially beetles of the genera, Dermestes and Necrobia throughout processing, transportation, marketing and storage. A survey in the market in Ibadan, Nigeria in the period from January 1971 to July 1972 showed that Coleoptera, especially Dermestes maculatus (which accounted for 71.5% of the observed infestation) and N. rufipes (28.0%), infested a high proportion of the dried fish sold (Osuji, 1974). Losses caused by D. maculatus infestation are enormous. They include physical loss whereby the amount of fish available for human consumption is reduced, economic loss whereby the physical loss despite the amount of fish available for sale and/or the price commanded for insect damaged fish is below that for undamaged fish, and nutritional loss, which is a direct consequence of the physical and economic losses and causes the retail value of fish to increase beyond the purchasing power of the poor (Moses, 1992). Although many synthetic chemicals are effective against the pests of many stored products, but efforts to reduce losses through insect infestation on smoke-dried fish by the use of insecticides and pesticides have not been fully adopted due to the hazardous nature of these chemicals to health and toxicity at high doses to users (Balogun, 1992); higher costs and less susceptibility of dermestid larvae (Amusan and Okorie, 2002; Odeyemi et al., 2000).
Several attempts have been made by man to control beetle infestation in cured fish and therefore mitigate the food, economic and health losses caused by them. Currently, worldwide interest is centered on the development of alternative strategies, including the re-examination of using plant derivatives against agriculturally important insect-pests. It was reported that when dried fish was mixed with leaf, bark, seed powder, or oil extracts of selected plants, there was mortality of beetles, reduced oviposition rate, suppression of adult emergence and reduced fish damage rate (Don-Pedro, 1985; Don-Pedro, 1989; Adedire and Lajide, 1999; Weaver and Subramanyam, 2000; Okonkwo and Okoye, 2001; Anyaele and Amusan, 2003).
The plant Phyllantus fraternus is a pan tropical weed which belongs to the family Euphobiaceae (Mehta et al. 2013). It is known for its hepatoprotective properties (Rastogi and Mahrotra, 1990). The plant is a common weed of cultivated fields; found abundantly in rainy season (Khan and Khan, 2004) and spreads widely in West Africa and other parts of the world where they are employed widely in traditional medicine preparations (Umoh et al., 2013). In Nigeria, it is known among Ibibios and Efik’s as ‘‘oyomokiso aman ke edem’’, Yoruba as “eyin olobe”, Hausa as “geeron tsutsaayee” and Igbo as “Ite knwonwa nazu” and in English as “leaf flower” or “chamber bitter” (Etukudo, 2003; Okujagu et al., 2005). Itis usually used as infusion and drunk by Nigerians for health maintenance and it is considered as a wonder- working herb and has great economic importance (Etukudo, 2000) but the insecticidal properties are not yet to the exploited.
Therefore this present study aimed at determining the phytochemical and larvicidal activity of powder of this medicinal plant in order to evaluate its insecticidal potential in suppressing infestation and damage by D. maculatus on stored smoke-dried fish.
1 Materials and Methods
1.1 Experimental location
The study was conducted at the Biology Laboratory in the Department of Science Laboratory Technology, Rufus Giwa Polytechnic Owo, Ondo State, Nigeria (7° 11' 43'' N 5° 33' 57'' E) under ambient condition of 30±2°C temperature, 65% relative humidity and 12L: 12D photo regime.
1.2 Collection and Preparation of leave powder
Whole plant of P. fraternus was collected from within the polytechnic community and the identity of the plant was confirmed at the Forestry and Wood Technology Department of Rufus Giwa Polytechnic, Owo, Ondo State, Nigeria. The plant material was shade dried for two weeks, and thereafter milled using an electric grinder and sieved to pass through 1 mm2 perforations.. The plant powder was kept in a separate sterile plastic container and placed in a cool dry place for use.
1.3 Insect culture and maintenance
The initial source of D. maculatus culture used for this study was obtained from natural infested smoked catfish (C. gariepinus) collected from smoked fish market stall in Owo, Ondo State, Nigeria. It was maintained in a kilner jar covered with muslin cloth under laboratory conditions and kept at a temperature 30+ 2 under relative humidity 65 + 5%. All bioassay jars was disinfected in an oven at 80 for 2 hours and were allowed to cool at room temperature. New generation was prepared by removing newly emerged (0-72h old) larvae from a stock culture, and placed on fresh uninfected fish, while the parent adult was removed after 2-3weeks oviposition period.
Smoked samples of the fish species (Clarias gariepinns) were obtained from smoked fish market stall at Oja oba market, Owo, Ondo State, Nigeria. The fish samples showed no visible presence of neither adult or larvae of D. maculatus infestation. The cured fish species were sterilized thermally by heating at 10 for one hour in a hot air oven (Gallenkamp Oven) in the laboratory in order to kill any insect pests that may be present (Atijegbe, 2004), and allow to cool at room temperature in the laboratory.
1.4 Effect of P. fraternus powder on larvae and adult emergence of D. maculatus
The toxic effect of P. fraternus on larvae D. maculatus was carried out using 250ml plastic containers containing 15g of smoked fish with concentration of 1.0g 2.0g, 2.5g and 3.0g for set A experiment, while set B experiment were admixed with concentration of 3.5g, 4.0g, 4.5g and 5.0g.The smoked fish in control dish contained no plant powder. The containers were gently shaken for 2 min to ensure homogenous mixture (Adesina et al. 2012) of the smoked fish and treatment powder. Ten newly emerged (0-72h old) larvae of D. maculatus were introduced into each treated and control dish and covered. Each treatment was in triplicate. Larvae mortality was counted every 24hours for 5days. The insects were confirmed dead when there was no response to probing with sharp pin at the abdomen (Adedire et al., 2011). Daily observations were made until adult emergence. The number reaching adult stages was recorded and percentage weight was also recorded. The percentage reduction in adult emergence of F1 progeny was calculated using the formula:
Percentage adult emergence reduction = 100 × (No. of adult insect emerged in control dish No. of adult insect emerged in treated dish) / No. of adult insect emerged in control dish.
The % loss in weight was determined and recorded using the method described by Odeyemi and Daramola (2000).
% Weight loss = 100 × (initial weight of fish sample final weight of fish sample) / initial weight of fish sample.
1.5 Phytochemical Analysis of P. fraternus
Sample of the plant leaf powder was chemically tested for qualitative phytochemical constituents using standard procedures recommended by Trease and Evans (1989); Harbone (1984); Mehta et al. (2013) and Umoh et al. (2013). For tannin, 5 g of each portion of plant extract was stirred with 10 ml of distilled water and filtered as described by Trease and Evans (1998). Blue black, green, or blue-green precipitate formed following the addition of few drops of 5% ferric chloride was taken as evidence for the presence of tannins. Deposition of a red precipitate when aqueous solution of leaf extract was boiled with 1% (v/v) HCl was taken as evidence for the presence of phlobatannin. Salkowski’s test, as described by Sofowora (1993), was used to test for cardiac glycosides. Leaf extract (0.5 g) was dissolved in 2 ml of chloroform prior to the careful addition of 1% (v/v) H2SO4 to form a lower layer. A reddish-brown colour at the interface was taken as evidence for the cardiac glycoside. Borntrager’s test was used for the detection of anthraquinones (Heyde et al., 1984). Plant material (5 g) was shaken with 10 ml benzene and filtered. Ammonia solution (5 ml, 10%) was added to the filtrate and a pink, red, or violet colour which was formed in the ammoniacal (lower) phase was recorded as an indication of the presence of free anthraquinones.
1.6 Experimental Design and Data Analysis
The experiment was laid out in Complete Randomised Design (CRD) and each treatment was replicated three (3) times. Data were subjected to analysis of variance and where significant differences existed, treatment means were separated using Least Significant Difference (LSD) at 5% probability level. Data in percentage were arcsine transformed before analysis.
2 Results
Table 1 shows the mortality rate of D. maculatus larva over a period of 120h after infestation. The results revealed that the plant powder exert significant (P<0.05) larva mortality with increase in application rate over time exposure; except for 24h after infestation in treatment A. In all the experiments dishes treated with 3g and 5g P. fraternus had the highest larval mortality, while fish protected with 1g and 3.5g had the lowest mean values of larval mortality respectively in both treatments.

Table 1 Percentage larval mortality of D. maculatus treated with different concentrations of P. fraternus powder

The larval mortality week after infestation was presented in Table 2. The results show that at a week after infestation (WAI) no mortality was observed in dish treated with 2.5g and 3g leave powder. Same for 2nd WAI and the mortality rate was not that significantly different (P<0.05) when compared statistically in treatment A (Table 2). While in treatment B there was significant difference (P<0.05) at 1st WAI with 5g treatment having the highest larval mortality (83.85%) and at 2nd WAI there existed no significant difference (P>0.05) among the treatments; although, highest mortality rate was recorded at 4g and 4.5g (Table 2).

Table 2 Percentage larval mortality of D. maculatus treated with different concentration of P. fraternus powder weeks after infestation

Weight loss as a result of the activities of D. maculatus showed a trend that reflected the number of surviving larvae and adult that emerged from the respective treatment dishes. However, the plant powder suppressed weight loss, although the observed differences were not significant statistically between the treatments.
The phytochemical analysis of the plant shows that the plant possesses the following constituents: Alkaloids, Saponins, Tannins and Kelakellani (glycosides) (Table 3, 4).

Table 3 Adult emergence of D. maculatus and percentage weight loss due to infestation on fish samples treated with different concentration of P. fraternus powder

Table 4 Results of Phytochemical Screening of P. fraternus Leaves Powder

3 Discussion
The use of botanicals for the control of insect pests of stored products is an ancient practice. Although this may be the first attempt to screen the plant for insecticidal properties and the result obtained from this study revealed that P. fraternus was found to be effective against larval stage of D. maculatus. The result is in agreement with many other researchers on the use of botanicals against suppression of D. maculatus infestation on smoked-dried fish (Onu and Baba, 2003; Adebote et al., 2006; Abdullahi et al., 2011; Abdullahi et al., 2012; Mufutau, 2012; Ahmed et al., 2013). The study clearly indicated that the higher dosage level of both treatments were the most effective in the application rates compared to the untreated control. The low adult mortality of F1 generation of D. maculatus could be as a result of high mortality of late instar larvae resulting from the toxicity effect exhibited by the plant powder. The emergence of F1 adult from all the treated dishes can also be attributed to the hairness of larvae which prevent direct contact of the powder on the body surface of the larvae as compared to the adult with smooth cuticle (Kemabota et al., 2013).
The higher percentage of weight loss recorded in untreated fish suggests that the larval stage of the beetle is more destructive than the adult (Nwankwo et al., 2011). Alam (2004) reported that if D. maculatus are left undisturbed, they can consume all the flesh and soft tissue of dried fish until only bone and some hard tissue remain. The larvicidal activity of P. fraternus may be attributed to the presence of bioactive constituents present in the plant. Rastogi and Mahrotra, (1990) reported that P. fraternus possesses following chemical constituents—phyllanthin, hypophyllanthin, niranthin, nirtetralin, phyltetratralin, kaempferol- 4- rhamnopyranoside and erio dictylol – 7- rhamnopyranoside etc. these bioactive agents could possess among other pharmaceutical properties, a depolarizing neuromuscular blocking action which could result to the death of insect (Udoh et al. 1999).
The study indicated that P. fraternus exhibited toxic effect against larvae of D. maculatus causing significant mortality and suppressed adult emergence and weight loss compared to control. The use of P. fraternus among poor resource fish farmers, processors and marketers should be advocated since the plant is widely distributed and used among rural folks for its ethno medical importance.
The technical assistance rendered by Miss Adeseila Omobola Comfort of Science Laboratory Technology Department, Rufus Giwa Polytechnic, Owo, Ondo State, Nigeria in collecting the data needed for this study is highly appreciated by the authors.
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