Survey Of Male And Female Frogs (Pyxicephalus Edulis) And Assosciated Toxic And Non Toxic Elements


 The present research was performed to determine the concentration of heavy metals in the male and female specimens of the identified frog species: pyxicephalus edulis collected from the Igbekebo River in Igbekebo, Ese-odo local government. Adult frogs (male and female) were collected from the river bank, and sediment samples and water samples were also collected at five ( 5) separate locations in the river. The frogs were dried separately at 1050C for 6 hrs and then crushed into small particles (powder form). The sediment samples were air-dried for three days. Elemental components in frog samples and sediment samples were analyzed using Proton Induced X-Ray Emission (PIXE). Physiochemical parameters and heavy metals of the water samples were also analyzed. The findings showed that the concentrations of Si, P, Cl, Ni, Zn and Cd were higher in Male frog while Mg, Al, K, Ca, Ti, V, Cr, Mn, Fe, Co, Cu, Zr, Pb and Sn were higher in female frogs, the explanation for this variability is not known but may be due to variations in the genetic make-up of Male and Female frogs. The concentration of heavy metals in both male and female frogs was substantially higher relative to the available WHO limits. The mean concentration of elemental constituents in sediment was higher than the IAEA limit. The values of enrichment and the Igeo values were very high.


Introduction
The worldwide decline in amphibian species has shown over-exploitation as one explanation for a decrease in global amphibian populations (Stuart et al., 2008). It has been said by Niasse et al ., ( 2004) that 281 amphibian species have the primary threat of utilization, 54% of which are already listed in their categories as vulnerable, endangered or critically endangered. A recent research by Warketin et al. ( 2009) sums up troubling data on Asian frog collected for use, with Mohneke (2011) reporting that a total of two million, 7800 and 610 frog species have been collected annually by 302 frog collectors in the southwest countries of Nigeria.A food scarcity and Africa behind other developed countries is one of the most apparent consequences of the world's population growth (Lameed, 2008).As a result, the production and supply of animal protein for feeding ever-expanding population decreased signi cantly (Akpan et al ., ( 2009).
Analysis was carried out with a 1.7 MeV 5SDH Pelletron Accelerator for sample metal concentrations. The pellets of sample were then analyzed for the total content of the elements (expressed as mg/kg) by the Particle-Induced X-ray Emission (PIXE) technique (He et al., 1993;Johansson et al., 1995) The technical detection limit is from 0.1 to 10 mg/kg. Table pH and conductivity (Hanna 991300) meters have been used to measure the pH and electric conductivity of the ground sample. The nitrate and chloride were also determined using standard procedures (Ademoroti 1996).Detailed analyses had been reported by (Ediagbonya and Balogun ,2020a; Bioaccumulation factor Bioaccumulation factor (BAF) explains the intake and distribution of the element in the organism after exposure in a given environmental matrix (Subotić et al., 2013); and has been used here to determine the level of frog with a higher risk to health (DeForest et al., 2007). The concentration of the sediment elements in the frog was measured as a ratio between the concentration of the frog element and the concentration in sediment (Liao and Ling, 2003;Javed and Usmani, 2013) and Bio-accumulation factor (BAF). BAF < 1 indicates no contamination of the frog; 1 > BAF ≤ 10, the frog is tolerant and BAF > 10, a hyperaccumulator (Ávila et al., 2017).
Health Risk Assessment Risk was assessed using the target hazard quotient (THQ) and the hazard index (HI). The risk to human health derived from the ingestion of contaminated food as speci ed by FAO / WHO (2010) and the target hazard quotient (THQ) as set out in the USEPA Region III Risk-based Concentration Table (USEPA , 2015). The THQ was calculated using the formula given by (Singh et al., 2010)THQ = Efr × ED × FIR × C / RfDo × Baverage wt × ATn × 10 − 3 Efr is exposure frequency assumed to be 365 days year − 1, ED is exposure duration in 54 years an average lifetime for Nigeria, while the average body weight used was 65 kg for this study (Oguntona,1998) FIR is average daily consumption taken as 1.95 x 10 − 2 kg person − 1 day − 1 C is concentration of metal in frog sample in mg/kg, RfDo i is the oral reference dose ( Al, Ti,V,Cr,Mn,Co,Pb, Cd, Cu, Zn, Fe andNi (USEPA,2011,2013 ;RAIS, 2017); and ATn is average exposure time for non-carcinogens and is taken as 19710 days. The hazard index (HI) from the consumption of frog estimated as the sum of THQs of all the metals in the frog and was expressed as follows; HI = THQAl + THQTi + THQV + THQCr + THQMn + THQFe + THQCo + THQNi + THQCu + THQZn + THQCd + THQPb Geo-Accumulation Index (Igeo).
The Geo Accumulation (Igeo) index is intended to measure the extent of sediment pollution as calculated by (Loska et al.1997;Müller, 1979). It was used by Igeo = In[Cn/1.5xBn] (Ediagbonya and Ayedun,2018;Ediagbonya and Balogun,2020). The calculated metal 'n' content in sediments is Cn and the background concentration of the same metal is Bn.

Enrichment Factor (EF)
To assess the extent of pollution in soil and sediments, the enrichment factor (EF) is used to determine excessive metal concentrations in sediments and soil. Al and Si were used as reference elements in this analysis. Few authors have used the reference feature of these components (Schiff and Weisberg, 199;Ediagbonya et al 2020b). The metal EF is classi ed as follows, according to Ergin et al., (1991): Where X/Al is the heavy metal concentration ratio (X) to the concentration of Al. Wedephol was taken from the reference crustal ratio of the shale value or lithology (1968) Statistical data analysis For the statistical analyses in this report, IBM Statistical Package for Social Sciences (SPSS) version 24.0 was used. Descriptive statistics at the various sampling sites, such as range, mean, standard deviation for the psychochemical parameters as well as heavy metals. One direction Variance Analysis was used to conduct the spatial variation of heavy metal means at the various sample sites where substantial difference was found to distinguish signi cant means was used by the Duncan Multiple Range Test (DMRT). The physicochemical parameters, using the Pearson correlation, were also associated with the toxic elements.The multivariate analysis conducted for the source detection of heavy metals was the principal component analysis. The signi cance standard was set at p < 0.055. Phosphate 8.71 ± 0.14 5.28 ± 0.14 5.12 ± 0.14 5.28 ± 0.14 5.12 ± 0.14 0.000 Nitrate 6.46 ± 0.14 6.52 ± 0.14 5.25 ± 0.21 6.52 ± 0.14 5.    living on the urban site in the Ukraine in comparisons between metal bioavailability in urban and rural frogs. Frogs are more sensitive to environmental toxins than other vertebrates because they do not allow foreign substances to be obstructed by egg membranes or skin (Duellman 1994;Snodgrass et al. 2003).
frogs have been used as bioindicators for pollution monitorisation, they have the proclivity to accumulate heavy metals (Berzins and Bundy 2002;Haywood et al. 2004;Simon et al. 2012). The consumption of frog is a result of Its availability, the taste for people and the need to meet the demand for protein are some of the key reasons for the annual consumption of billions of frogs. Frogs are also collected for leather production and souvenirs, pet trade and cultural purposes, including traditional medicine (Kusrini and Alford, 2006;Gonwouo and Rodel, 2008). From Table 4  The mean comparison of metals in male and female frogs is shown in Table 5. There was a signi cant difference (p < 0.05) in the mean concentrations of Al, Cl, Fe, Cu and Zn, while the other elements did not show any signi cant differences (p > 0.05). In Al, Fe and Cu, females had higher concentrations than males, while in Cl and Zn, males had higher concentrations than females.