See discussions, stats, and author profiles for this publication at: https://www.researchgate.net/publication/340226118 Detection of cyanide in freshwater ﬁshes relative to sex dimorphism using landmark-based geometric morphometrics in Agusan del Sur, Philippines Research · February 2018 DOI: 10.12692/ijb/12.2.177-193 CITATION READS 1 908 1 author: Rex Saura Caraga State University 6 PUBLICATIONS 3 CITATIONS SEE PROFILE Some of the authors of this publication are also working on these related projects: DETECTION LEVELS OF CYANIDE IN FRESHWATER FISHES RELATIVE TO THEIR BODY SHAPE VARIATION USING LANDMARK-BASED GEOMETRIC MORPHOMETRICS IN BUNAWAN, AGUSAN DEL SUR View project All content following this page was uploaded by Rex Saura on 27 March 2020. The user has requested enhancement of the downloaded file. Int. J. Biosci. 2018 International Journal of Biosciences | IJB | ISSN: 2220-6655 (Print), 2222-5234 (Online) http://www.innspub.net Vol. 12, No. 2, p. 177-193, 2018 RESEARCH PAPER OPEN ACCESS Detection of cyanide in freshwater fishes relative to sex dimorphism using landmark-based geometric morphometrics in Agusan del Sur, Philippines Rex Bomvet Deleon Saura*1,3, Rex Jr. Masinading Andante2,3 1 Surigao State College of Technology-Mainit Campus, Surigao del Norte Philippines 2 DepEd-Senior High School Teacher, Bunawan National High School, Bunawan Agusan del Sur, Philippines 3 Graduate Student, College of Education, Caraga State University, Ampayon, Butuan City, Philippines Key words: Cyanide, Physico-chemical, Freshwater fish, Sex dimorphism, Allometry. http://dx.doi.org/10.12692/ijb/12.2.177-193 Article published on February 26, 2018 Abstract This study was done for public health and environmental concerns and aimed to determine the presence of cyanide in three fishes such as Channa striata, Oreochromis niloticus and Clarias batrachus, and described the allometry and sex dimorphism of Chevron Snakehead. The physico-chemical parameters, levels of Cyanide, Copper, Lead, Zinc and Iron in nine (9) stream tributaries of Agusan Marsh was evaluated and compared to the DENR freshwater quality standards. The research employed Cyanide Detection Tests, length-weight relationship, land-marking and relative warp analysis using tps software and PAST-Hotellings test. The result revealed that 82% were positive in cyanide, found in all two (2) samples O. niloticus, in C. batrachus four (4) is positive only one is negative, three (3) C. striata, were positive and only one (1) is negative. Negative allometric were observed to 15 male and 15 female Channa striata showed b value is 1.0798, strong positive correlation with r2 = 0.8908 and significant with p=0.00001. No shape variation among male and female C. striata . Hotelling’s test where T2 = 6.168, F = 1.1774, P = 1, means no signiﬁcant sexual dimorphism for the body shape of the C. striata. Moreover, lead, copper, zinc and iron and cyanide in water revealed in detectable levels and physico-chemical water quality of the streams were affected.Thus, landmark based geometric analysis were useful in describing sexual dimorphisms among fish species. In addition, freshwater fishes were sensitive and susceptible to bio accumulation of cyanide and growth affected by the water quality. * Corresponding Author: Rex Bomvet Deleon Saura email@example.com 177 Saura and Andante Int. J. Biosci. 2018 Introduction Furthermore, cases of cyanide leak from mining in Caraga Region have soundly ecosystem dome for Bunawan, Agusan del Sur had been reported in the organisms to live. “It is one of the 128 key biodiversity newspaper (Enerio, 2017). Thus, this study was area and the top 10 priority protected areas in the conducted and evaluated the presence of cyanide in Integrated Protected Areas System, abundant in water and three (3) selected common fish commodity fishes and endemic species of amphibians and such as Chevron Snakehead (Channa striata), Nile reptiles” (Ngilangil et al., 2014). Tilapia (Oreochromis niloticus) and catfish (Clarias Pertaining to industrial mining, Caraga Region, Philippines has several registered large scale mining companies based on the data from Mines and Geosciences Bureau-Surigao as of 2015, not only large batrachus) including base metals and its physicochemical analysis in nine (9) selected stream tributaries of Agusan Rivers. In addition, this study attempted to described the morphology of the scale mining but active small scale mining operations Chevron Snakehead (Channa striata) as to allometry also existed region wide. Since 1970’s, cyanide widely and compare its shape variations in relation to their used in the gold mining industries to harvest gold but sex a minute sized amount of cyanide can be deadly to landmark based using morphometrics software. differences using geometric morphometric humans and even smaller amounts can be lethal to fishes once contaminated water (Mineral Policy This provide significant information on the Center, 2000). occurrence of four base metals, cyanide and water quality of stream tributaries of Agusan Rivers and Impact of cyanide to aquatic organisms like fish lost accumulation of cyanide in the fish tissue of Chevron its equilibrium with too much mucous discharge on snakehead, Nile tilapia, and catfish samples which its gill filaments and skin (Ramzy, 2014). Increase included the allometry and body shape variation of water temperature, minor reduction of pH and Channa striata relative sex differences and provide dissolved oxygen, disturbs fitness and stress to fishes awareness for the environmental and public health (BFAR, 2014). Freshwater fish were the most cyanide-delicate group of aquatic organisms tested, with high mortality and adverse effects on swimming and fecundity (Eisler and Wiemeyer, 2004). Wildlife death likely occurs near mines processing copper gold due to formations of copper-cyanide complexes which is toxic to birds and bats (Donato et al., 2007). sectors, mining companies and local government for monitoring and formulation of policy and regulation to minimize the entry of toxic pollutants into water bodies. Methods and materials Study area In general, water pollutants like decayed plants can affects fish body growth and shape because of biological oxygen demand in decomposition process lowers dissolved oxygen level in water resulting fish ability to survive. Similarly fish accumulation of toxins can be transfer to other trophic level as they are eaten (William, 2017). Cyanide instigated severe initial The study was conducted in the Municipality of Bunawan, Agusan del Sur, Philippines shown in Fig. 1 with their respective coordinates. The ten (10) sampling stations for water samples was Magsagangsang Creek, Lucad Creek, Simulao-lower stream, Bunawan Brook River, Saugon creek, Bagnan suppression of specific growing proportion (Dixon & creek, Simulao River-upper stream , Singanan Creek, Leduc, fishes Koo Creek and Masapia falls, within three months contaminated with metals was impaired sense of smell, duration started from September to November 2017. resulting to lessen from escaping predator that the The collection of samples and field sampling proper problem could distress fish density (Bienkowski, 2013). was done once only. 1980). American study 178 Saura and Andante in lake Int. J. Biosci. 2018 Fig. 1. Map of study stations with coordinates in Bunawan, Agusan del Norte. Establishment of sampling stations The samples were carefully transferred to the The collection of water samples together with physic- prepared chemical parameters was done in nine (9) streams terephthalate) plastic bottles of nature spring to avoid and only one sampling stations in each stream but bubbles formation. The bottles was sealed tightly and except for Simulao river with two (2) station was labeled with information as to area, station, and date established upper and lower streams. A total of ten of collection and photograph for documentation. (10) sampling area was established. Establishment of Then all samples were placed in the bucket with sampling was only in the accessible portion of the splitter and ice for preservation. river. On the other hand, the collection of fish samples was obtained from local fisher folks of Bunawan, Agusan del Sur. Philippines. Collection and sampling procedures for water analysis Collection procedures for water samples was adopted and modified from https:www.env.go.jp>04- Guideline/Chapter 2 sampling and Veiga and Baker, 2004. The water samples were collected through depth of 1-50 cm of the water column of each sampling sites. About 0.5 L of water samples only reused 0.5 L PET (polyethylene Collection, data gathering and Preparation for fish biota samples Freshwater fish samples such as Nile Tilapia, chevron snakehead and Catfish was obtained from the local fisher folks Bunawan, Agusan del Sur. About 1.3 kilogram of each fish types was obtained as prescribed by the BFAR 13 for cyanide analysis and another thirty (30) samples of Chevron snakehead for landmark based geometric morphometric analysis were obtained. All Fish samples were washed with tap water then after were rinsed with distilled water. was collected in each stations using graduated The weight of each fish samples of three (3) species cylinder, a total of ten (10) bottles with 0.5 L water was determined using digital weighing scale. All samples each with only one replicate. The graduated samples for cyanide analysis were subject to cylinder will be rinsed with distilled water four times photograph for documentation using iPhone 5s before using over again. camera. 179 Saura and Andante Int. J. Biosci. 2018 Hereafter, the fresh samples of fish was placed in a Shape analysis zip lock and labeled with information as to area, The saved files of digitized samples were analyzed station, and date of collection and placed in the through TPS relative warp where Relative warp score bucket with ice and frozen. Moreover, the thirty (30) and deformation grid and PCA retrieved. Further Chevron snakehead sample collected for geometric analysis for sexual dimorphism was done using PAST morphometric analysis were separately placed in a software bucket with ice for geometric morphometric analysis. generations of histogram and discriminant plot. Analytical procedure for cyanide and base metals Data analysis and consolidation of data collected The collected water samples were immediately Ecological aspect of the sampling was described brought to Philsaga Mining Corporation, Assay through ocular and observation to the ecological Laboratory for Cyanide and Base metals Analysis. profile for of the the discriminant sampling sites. analysis The and allometry determination was determined using the equation The prepared fish samples were delivered to W=aL b (Pauly, 1983). At Philippine Bureau of Fisheries and Aquatic Resources where : (BFAR) Central Office-Manila thru BFAR Caraga for W= weight of fish (g) the cyanide detection and analytical procedure. The L = total length of fish (cm) physico- chemical analyses were done using the Hach a = constant (intercept) HQ40D. b = the length exponent (slope) Fish sample preparation for Land-marking The prepared fish samples the length of each fish samples was determined by measuring the fish from its snout down to the caudal fin using tape measure and weighed using digital weighing scale. The length and weight were recorded. The individual fish was applied with 10% formaldehyde to harden the fins and manual manipulation was done to make it more visible and wide. values of Riedel et al.,2007 presented in Table 2 and Pearson Correlation was used for r2 values and coefficient of determination with their corresponding interpretation shown in Table 3. Furthermore, The results of the cyanide analysis was analysed as negative or positive only and the concentration of cyanide in water of each sampling were compare only if it is above or below to the maximum contamination level. The left flank of each fish taken using cannon eos 1200 D for land-marking. The result was interpreted using linear regression Samples was dissected individually to determine the sexes and the sorted as to male and female. Land-marking selection and digitization The images of fish sample of each sex were sorted and replicated into two (2). Then the images were subjected to tpsUtil64 to create tps files images for digitization. A total of 16 landmarks were digitized using tpsDig2 software. The location of the landmarks and anatomical descriptions of each sex were represented in Figure 2 and Table 1. Assigning landmark were based on stickleback fish model of Anyfish.com. 180 Saura and Andante Results and discussion The sampling area were surrounded of agricultural land like rice fields, rubber tree and falcate plantation, compose of shrub to trees and coconut. East areas of Bunawan observed high to low range of mountains with perceived landscape of mining activities and during the actual survey fishing were observed. The fish caught by local fisher folks either for family consumption. Allometry The thirty (30) individual samples of chevron snakehead (Channa striata) were sorted by sexes and measured the total length and then weighed and recorded. Int. J. Biosci. 2018 The length ranged from 26.0 cm – 37.5 cm with 32.6 weight ranged from 218.66 g - 315.37 g with 273.7 g cm average mean for male while female ranged from average mean for male while 243.89 g – 320.55 g for 29.0 cm to 35.5 cm with 32.3 cm average mean. The female with 284.2 g average mean. Table 1. Description of the landmark points in the body shape of Channa striata. Coordinates 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 Location Snout Tip End of head length anterior junction of the dorsal fins Posterior junction of the dorsal fins Dorsal insertion of caudal fin Midpoint of caudal border of hypural plate Ventral insertion of caudal fin Posterior insertion of anal fin Anterior insertion of anal fin Dorsal base of pelvic fin anterior border of ectocoracoid bon anterior border of ectocoracoid bon Anterior margin through midline of orbit Posterior margin through midline of orbit Dorsal end of operculum Dorsal base of pectoral fin The results of Length- weight relationship of fifteen as shown in Fig. 4. The graph shown that b value (15) male and 15 female Channa striata collected in result is 1.0798 which means negative allometric, the month of October were and presented in graph as correlation coefficient or r2 is 0.8908 means strong shown in Fig. 3. positive correlation, p value is 0.00001 which means that there is significant association of length of the Furthermore, the length and weight relationship of collected samples. combined sexes of Channa striata were determined Table 2. Corresponding interpretation of the b values for linear regression (Riedel et al, 2007) b value Interpretation. b value Interpretation 3.0 <3.0 >3.0 Isometric growth- no change of body shape as an organism grow Negative Allometric- body shape more slender as it increase in length Positive Allometric- - body shape relatively stouter as it increase in length Table 3. The values of (r2) coefficient of determination with their corresponding interpretation. Coefficient of Determination ((r2) +.70 or higher +.40 to +.69 +.30 to +.39 +.20 to +.29 +.01 to +.19 0 -.01 to -.19 -.20 to -.29 -.30 to -.39 -.40 to -.69 -.70 or higher 181 Saura and Andante Interpretation Very strong positive relationship Strong positive relationship Moderate positive relationship weak positive relationship No or negligible relationship No relationship [zero order correlation] No or negligible relationship weak negative relationship Moderate negative relationship Strong negative relationship Very strong negative relationship Int. J. Biosci. 2018 In general, all samples of Channa striata data length-weight revealed negative allometric which means that the significant difference amid male and female Channa growth of fish become slender as it is increase in striata and in terms of their condition factor was poor length. revealed indicated 1.50 and 1.49 for and female respectively. significant associations; this entails that as length (Dumalagan et al., 2017). Another study of Channa increase the weight of fish also increase but in more striata collected in different river results showed that slender. Similar studies on the allometric growth was in Ganga River displayed isometric growth b equal to observed in chevron snakehead tributaries of Lake 3.012, in Yamuna b is 2.18 and in Gomti b value is Kilobidan, Agusan Marsh, Philippines in terms of 2.86 revealed a negative allometric shape of growth. The correlations coefficients relationship that there was no Table 4. Parameter concentration level of 10 stations collected last November, 2017. Parameters Stations Magsagangsang Creek Lucad Creek Simulao River-(lower) Bunawan Brook River Saugon creek Bagnan creek Simulao River-(upper) Singanan Creek Koo Creek Masapia Falls DO ppm 7.5 6.9 5.4 5.3 4.8 6.0 7.3 5.1 6.1 6.2 *pH 6.25 6.36 6.32 6.63 6.72 6.5 6.48 6.6 7.0 6.75 Saline ppm 500 620 1350 1590 1700 480 131 750 570 550 TDS Ppm 503 618 1335 1569 1674 494 1292 746 564 551 *TSS ppm 9.8 21 13.8 16.8 24.2 15.6 22.4 12.6 9.2 17.8 Conductivity μS/cm 1018 1245 2617 3060 3250 1003 2541 1494 1147 1113 Resistivity Ω⋅m 980 802 382 327 307 997 393 668 877 895 *Results given by Philsaga Mining Corp. Assay Lab of Bunawan, Agusan del Sur, Philippines. Table 5. Cyanide and Four Base Metals concentrations of ten (10) stations collected last October, 2017. Stations Magsagangsang Creek Lucad Creek Simulao River-(lower) Bunawan Brook River Saugon River Bagnan River Simulao River-(upper) Singanan Creek Koo Creek Masapia Falls *< presence. CN(ppm) <0.01 <0.01 0.01 0.01 0.01 0.01 0.02 0.01 0.01 <0.01 Cu (ppm) 0.02 0.01 <0.01 0.01 <0.01 0.01 <0.01 <0.01 <0.01 <0.01 Pb (ppm) 0.02 0.15 0.02 <0.01 <0.01 <0.01 0.02 0.02 0.01 0.02 Base Metals Zinc (ppm) 0.02 0.03 <0.01 <0.01 <0.01 <0.01 0.01 <0.01 <0.01 <0.01 Iron (ppm) <0.01 2.32 0.41 0.41 0.47 2.34 0.3 1.05 0.43 0.19 In addition r2 was 0.99, 0.94 and 0.97 from different Fish allometry affected by fish habitat and nutrition river respectively which length and weight were availability in the area. highly correlated (Khan, 2014).) Length-Weight Relationship of Channa striata is isometric b value is 3.0 and larger individuals having negative allometric relationship b less than 3, represents possible agelinked variation in growth form (Ali et al., 2013). 182 Saura and Andante Sexual dimorphism Relative warp analysis using tpsRelw ver.1.69 was used to determined differences in shape through deformation grid among specimens in term of sexes. The sample does not include pooled data analysis. Int. J. Biosci. 2018 Table 6. Range and Comparison of parameters concentrations to DENR standards. Parameters Present Data DENR DO, ppm pH Salinity, ppm TDS, ppm TSS, ppm Cyanide , ppm Cu , ppm Pb, ppm Zn, ppm Fe, ppm Conductivity μS/cm 4.8--7.50 6.25 --7.0 500 -- 1700 503—1674 9.2--24.20 <0.01-- 0.02 <0.01-- 0.02 <0.01-- 0.15 <0.01-- 0.03 <0.01---2.34 1003---3250 5 6.5-9.0 Nd 96-100 80 0.1 0.02 0.02 1.5 2 Nd 307---997 Nd Resistivity Legend : < presence , nd- no data Table 7. CDT results of fish samples positive in cyanide. FISH N Test Parameters Test Methods Specifications Results Nile Tilapia 2 Cyanide ISE-APHA 4500 positive Chevron Snakehead 3 Cyanide ISE-APHA 4500 Limit of Detection 1.209 E-2 mg/Kg Limit of Detection 1.209 E-2 mg/Kg Catfish 4 Cyanide ISE-APHA 4500 Limit of Detection 1.209 E-2 mg/Kg positive positive N-total number of samples. The PCA of each sex were shown in Figure 5 and Based on the cumulative report retrieved relative Figure between warp analysis, both of the specimen male or female deformation grid of the negative relative warp which have the same percentage of Principal components., is on the left side and positive relative warp on right PC1 is 33.15%, PC2= 21.98 %, PC3=11. 35% , side of each Principal Component. The shape PC4=8.56 %, and PC5 8.08% as shown in Fig. 5 and 6 variation is less among species which means that respectively. 6 which include Histogram shape differences also less base on 16 landmarks. Table 8. CDT results of fish samples negative in cyanide. Fish Samples N Test parameters Test Methods Specifications Results Chevron Snakehead 1 Cyanide ISE-APHA 4500 negative Catfish 1 Cyanide ISE-APHA 4500 Limit of Detection 1.209 E-2 mg/Kg Limit of Detection 1.209 E-2 mg/Kg negative N-total number of samples. The mean thin spline models illustration as express Furthermore, discriminant analysis using PAST by landmark showing the same grid formation of software revealed that there is no sexual dimorphism sixteen (16) anatomical land mark points of male and between male and female as presented in Fig. 8. The female as shown in Fig. 7. This implied that no graph showed the discriminant plot that overlaps anatomical landmark points were affected. which mean same shape differences between male 183 Saura and Andante Int. J. Biosci. 2018 and female Channa striata sample. Hotellings T2 test both in T. truncatus (Guidarelli et al., 2014). The results revealed no signiﬁcant sexual dimorphism for important difficult concerns why the sexes are the body shape of the Channa striata Hotelling’s T2 = frequently so alike in size this might tell to sperm 6.168, F = 1.1774, P = 1. competition, size buffering effects, and many other features (Parker, 1992). However, Chevron snakehead Similar result of sexual dimorphism for the shape has female are larger than male according to Chaudry of no significant or shape were the same of the mandible 2010. Fig. 2. Landmark points of female (A) and male (B) Channa striata. Fig. 3. Split Scattered Plot Diagram of Log Total Length and Log Total weight of Male and Female Channa striata. 184 Saura and Andante Int. J. Biosci. 2018 In general, negative allometry was observed in the Agency for Toxic Substances and Disease Registry samples that as fish grow in length it’s become provide results of the toxicity level Cyanide and its slender. Some factors may affects fish growth effects and fund out that, at 0.05 mg/dL is toxic to according to Tubb 1972, cyanide had various distinct blood, 0.3 100 and higher were deadly while in special effects on growing cichlids, outcome in erratic hydrogen cyanide at 546 ppm with 10 minutes time exposure and 110 ppm after one hour exposure are of contact and with the level tried cyanide upheld reduction in fish growth. Cyanide can both lethal. disturbs fitness and stress to fishes (BFAR, 2014). Fig. 4. Scattered Plot Diagram of Log Total Length and Log Total weight of Channa striata. Fig. 5. Five (5) Principal components (PC) implied grid and histogram of individual shape in Channa striata male species. 185 Saura and Andante Int. J. Biosci. 2018 At the same time, people who eat small amounts of of enormous phenotypic malleability evolutionary cyanide compounds in a short time may decease characters relative ecological physiognomies and this unless they rapidly take remedy therapy. Sex was studied on the fish Mesopristes cancellatus in dimorphism general Tagoloan River using tps relwarp revealed significant characteristics of external appearance, shape and sex dimorphism variation of body shape based on sizes among species Fishes were recognized on behalf landmark is the differences in generated (Leopoldo et al., 2014). Fig. 6. Five (5) Principal components (PC) implied grid and histogram of individual shape in Channa striata female species. The software was very useful in learning intraspeciﬁc Physico-chemical, cyanide and four based metals variation, or perhaps to examine micro-evolution or The water parameters such as Dissolved Oxygen, pH, distinguish among kind which may be interpreted in Salinity, Total Dissolve Solids, Total suspended terms of environment, genetic differences, sexual dimorphisms, instars or year classes and Principal components analysis (PCA) is a method that produces solids, conductivity and resistivity were determined presented in Table 4. The cyanide, Copper, Lead, Zinc hypothetical variables or components, accounting for and Iron of the water samples from 10 stations as much of the variation in the data as possible shown in Table 5. The results were compared to (Hammer, 2002) Wider uses of these ﬁndings can DENR standards as presented in Table 6. During the link to evolutionary biology, organization, and sampling proper water temperature were observed preservation (Bell and Jacquemin, 2017). Landmark- from 25 o C to 28.5 o C in the sampling stations. From based geometric morphometrics is a powerful approach to quantifying biological shape, shape variation, and co-variation of shape with other biotic the Table 4, the range of DO was 4.8 ppm of Saugon creek to 7.50 ppm of Magsagangsang creek. The pH or abiotic variables or factors. The resulting graphical level ranges from 6.25 and 7.0 of Bagnan creek and representations of shape changes were visually Koo creek respectively. attractive and instinctive (Webster and Sheets, 2010). 186 Saura and Andante Int. J. Biosci. 2018 Fig. 7. Mean thin plate spline models illustrating the same grid of Male and Female Channa striata. The salinity is ranging from 500 ppm observed from Magsagangsang creek to 3250 μS/cm which observed Magsagangsang creek to 1700 ppm recorded from from Saugon creek with minimum resistivity which Saugon creek. The range of TDS is 503 ppm to 1674 307 Ω⋅m and high resistivity of 997 Ω⋅m observed in ppm Magsagangsang and Saugon respectively. The Magsagangsang creek. conductivity is ranging from 1003 μS/cm recorded in Fig. 8. The discriminant plot of sexual dimorphism of Male and Female Channa striata. The result of the Laboratory analysis of all water positive samples in terms of cyanide and base metals concentrations from <0.01 to 0.02 ppm. Less than concentrations was presented in the Table 5. Based on 0.01 (<0.01) it doesn’t mean not detected but the report and result, all waters samples from the ten interpreted as presence of cyanide. On the other (10) stations of nine (9) stream tributaries were hand, the four base metals have the lowest 187 Saura and Andante of cyanide but varied in detectable Int. J. Biosci. 2018 concentrations of <0.01 ppm. Copper recorded upto while salinity, total dissolved and suspended solids 0.02 ppm of Magsagangsang creek. Lead and zinc top were the highest and exceed the acceptable level as concentrations observed at Lucad creek and Bagnan well as the conductivity was the highest inversely to river observed with 2.34 ppm Iron. resistivity relatively low compare to other area. However, all sampling area reached beyond the Furthermore, the water parameters such as Dissolved normal level of total dissolved solids. pH 6.25 ppm in Oxygen, pH, salinity, and total Dissolve Solids, Total Magsagangsang creek deviates minute level of pH to suspended resistivity. the accepted value. In Lucad creek the Lead (0.15 Including cyanide, Copper, Lead, Zinc and Iron were ppm) and Iron (2.32 ppm) exceed above acceptable compared quality and also the Iron (2.34 ppm) level in Bagnan. Cyanide standards as shown in Table 6. Comparison revealed and Copper and Zinc ranged from below detectable that Saugon dissolve oxygen was lower at 4.8 ppm, and equal to the accepted value. solids, with conductivity the DENR and freshwater Fig. 9. Percentage of Fish samples positive in cyanide. Total dissolved solids is relatively high and exceeded concentration level of cyanide and other four base DENR standards because the day before sampling metals or two hand way either diluted due increase in there was low pressure and weather conditions were volume or more added other metal complexes. The cloudy to rainy during sampling. High runoff of four base metals like Pb and Cu is a primary pollutant sediments due to wet weather conditions contribute while Zn and Fe were secondary pollutants that to high dissolved and suspended solids in fact water should be monitor otherwise adverse effect colour to in most area were turbid through ocular observation water and organisms. Lead should be 0.015 mg/L except in Masapia Falls. The turbidity it is a relative maximum concentration limits because potentially increase of TDS, TSS, (Fondriest Environmental Inc., affects metal development to infants and children, 2014) which also increase salinity and conductivity, deficits attention and learning, for adults it is kidney this was observed from the water sample. The target related problems (US EPA, 2017). analytes which is cyanide is very low in concentration observed may be because of the rainy weather Cyanide detection in fish sample conditions since it falls to wet season in the country Three (3) freshwater fish species such as Nile Tilapia that rises water level in water and affected the weighed 1,147.54 g with two (2) individual, Chevron 188 Saura and Andante Int. J. Biosci. snakehead weighed 2018 (4) In this study, landmark based geometric analysis individual and catfish weighed 1358.78 g with five (5) 1100.08 g with four revealed no shape differences of male and female individuals collected in the month of October with a Channa striata and also observed negative allometric total of 11 fish sample was sent to BFAR Manila for growth which so far accumulated cyanide as well as Cyanide Detection Test or CDT. The analysis used Ion other fish samples such as catfish and Nile tilapia due Selective Electrode reference to American Public to cyanide contamination of water from nine (9) Health Agency methods no. 4500 with a Limit of stream tributaries of Agusan Marsh. This implies that Detection 1.209 E-2 mg/Kg and results revealed that fishes greatly affected water quality and other Cyanide were found positive in all Tilapia samples, in ecological disturbance. Similarly, an analysis on Ibis three (3) samples of chevron snakeheads and in four fish (Ambassis interrupta) collected from Masao (4) Catfish as shown in Table 7. River, Butuan City, Philippines revealed the two (2) factors sides and interaction had significant In Table 8 showed the fish sample negative with differences with (P<0.0001) which influence by cyanide, one (1) in chevron snakehead sample and prevailing factors like existence of fish ponds, use of one (1) Catfish sample. feeds and publics home lengthways Masao River affecting water chemistry of river that might also Furthermore, the result revealed that 82% were positive in cyanide,18% is negative shown in Figure 9. Accordingly, “positive means cyanide (CN-) concentrations in fish is greater than 0.20 mg/L. Traces means cyanide (CN-) concentrations from 0.05 to 0.20 mg/L. Cyanide levels greater than zero (0) distress general phenotype of the fish (Medrano and Jumawan, 2016). Fluctuating asymmetry of fishes were susceptible near ecological changes that possibly manifested the size and forms which analyzed by utilizing Geometric Morphometric which vital in recognizing body differences and similarities of species in the same taxon (Berame et al, 2017). and less than 0.05 mg/L are scored BDL-below detectable limits. However, zero (0) values are Conclusion considered to be negative” (BFAR, 2001).While most Therefore the study concluded that pollutants such prosecutions deal with fish that tested as being lead, copper, zinc and iron and cyanide were already Positive, any level of cyanide found in the fish is an in detectable levels and contaminated water quality in indication that the fish were exposed to cyanide stream tributaries of Agusan Marsh. Further, land- coming from anthropogenic sources. mark based geometric morphometric were a useful tool in comparing the shape variation of Channa This indicates the fish samples accumulate striata relative to sex dimorphism. The relative warp detectable concentration of cyanide from water since analysis cyanide was also detected in water samples. Fishes component are aquatic differences with p=1 which means the body shape of organisms (Eisler and Wiemeyer, 2004). In water male and female were same in shape likewise cyanide can rise water temperature, minor reduction provided a strong positive correlations of length – of pH and dissolved oxygen disturbs fitness and weight relationship and both sexes have negative stress to fishes (BFAR, 2014). There are reports that allometry. Also, a positive accumulation of cyanide in even cyanide concentrations as low as 0.01-0.1 mg/L Channa striata, Oreochromis niloticus, and Clarias lethal to sensitive animals species present in waters. batrachus samples were observed thru cyanide Sodium cyanide (NaCN) evidently accumulated to detection tests. Thereby bio accumulation of cyanide most cyanide-delicate group of water species and absorption factors for various aquatic organisms (International Management Code, 2015). 189 Saura and Andante Cyanide similarly provided analysis first revealed five no principal significant pertains to the detectable levels of cyanide and growth of fish affected by primary and secondary pollutants such as base metals in the fish habitat. Int. J. Biosci. 2018 This study suggest to LGU in the monthly monitoring Bienkowski B. 2013. Environmental Health News as to cyanide analysis in water and fish and provide Fish Cannot Smell in Polluted Waters Retrieved strict implementation of cyanide waste disposal in November 13, 2017. mining industries. www.scientificamerican.com/article/fish-cannotsmell-in-polluted-waters/ References Ali A, Dahanukar N, Raghavan R. 2013. Lengthweight and length-length relationship of three species of snakehead fish, Channa diplogramma, C. marulius and C. striata from the riverine reaches of Lake Vembanad, Kerala, India. Bureau Of Mines And Geoscience–Surigao. 2015. Mining Industries. Retrieved October 13, 2017 from http://mgb.gov.ph/transparency/2015-05-04-07-0356/2015-05-04-07-57-31 http://dx.doi.org/10.11609/JoTT.o3353.4769-73 Anyfish. 2016. Modeling body and fin shape Chua K. 2015. TAXO4254: Channa striata Retrieved (morphometrics). Retrieved October 13, 2017 from November 13, 2017 from http://swordtail.tamu.edu/anyfish/Modeling_body_ https://taxo4254.wikispaces.com/Channa+striata. and_fin_shape_(morphometrics) Courtenay W, Williams J. 2004. Snakeheads Agency For Toxic Substances And Disease (Pisces, Channidae)—A Biological Synopsis and Risk Registry. 2015. Toxicological profile for Cyanide. Assessment. Retrieved November 13, 2017 from Atlanta, GA: U.S. Department of Health and Human https://pubs.usgs.gov/circ/2004/1251/report.pdf Services, Public Health Service. www.atsdr.cdc.gov/PHS/PHS.asp?id=70&tid=19 Bell J, Jacquemin morphological and J. 2017. functional Chaudhry S. 2010. Channa striata. The IUCN Red Evidence variation of among Bluegill Lepomis macrochirus populations across Grand Lake St Mary's watershed area, Journal of Freshwater Ecology 32(1), 1-18. http://dx.doi.org/10.1080/02705060.2017.1319429 List of Threatened Species 2010: e.T166563A6237224. Retrieved November 13, 2017. http://dx.doi.org/10.2305/IUCN.UK.20104.RLTS.T1 66563A6237224.en DAO-DENR, PHILIPPINES. 2016. Water Quality Guidelines and General Effluent Standards of 2016. Berame J, Busia M, Nova D. 2017. Describing Retrieved November 18, 2017 from body shape variation of bugwan fish (Hypseleotris http://server2.denr.gov.ph/uploads/rmdd/dao-2016- agilis) species using land mark-based geometric 08.pdf morphometric in Lake Mainit, Surigao del Norte, Philippines Journal of Biodiversity and Environmental Sciences 11(3), 113-121, 2017. Dixon G, Philippines. 2014.Cyanide. Retrieved October 13, 20 from www.bfar.da.gov.ph/cyanide.jsp BFAR, Philippines. 2001. Standard Operating Procedures For Cyanide Testing Used By The G. 1980. Chronic cyanide poisoning of rainbow trout and its effects on growth, respiration, BFAR, Leduc and liver histopathology Retrieved November 18, 2017 from https://link.springer.com/article/10.1007/BF010575 80 Donato B, Nichols O, Possingham H, Moore Philippines Cyanide Detection Test (Cdt) Laboratory M, Ricci Pf, Noller B. 2007. A critical review of the Network. Retrieved March 09, 2017 from effects of gold cyanide-bearing tailings solutions on http://ftp.nodc.noaa.gov/pub/data.nodc/coris/.../SO wildlife. P_CyanideTesting_CDT.pdf www.ncbi.nlm.nih.gov/pubmed/17540.445 190 Saura and Andante Int. J. Biosci. 2018 Dumalagan F, Garcines J, Boyles L. 2017. IT IS. 2015, Integrated Taxonomic Information Reproductive Biology, Length-Weight Relationship System: Channa striata. Retrieved on November 13, and Condition Factor of Channa striata (Bloch, 1793) 2017 from from tributaries of Lake Kilobidan, Agusan Marsh, www.itis.gov/servlet/SingleRpt/SingleRpt?search_to Philippines pic=TSN&search_value=166667.null Int'l Journal of Computing, Communications & Instrumentation Engg. (IJCCIE) 4(1). International Cyanide Management Code, 2015. Environmental & Health Effects. Online: http://dx.doi.org/10.15242/ijccie.ae0117114 February 14, 2017. Enerio B. 2017. EMB To Go After Miners Using www.cyanidecode.org/cyanide-facts/environmental- Banned Chemicals Retrieved October 13, 2017 health-effects www.sunstar.com.ph/cagayan-de- Khan S. 2014. A comparative study on length-weight oro/business/2017/09/08/emb-go-after-miners- relationship, length-length relationship and condition using-banned-chemicals-562914 factor of a freshwater snakehead, Channa striata from Eisler R, Wiemeyer S. 2004. Cyanide hazards to plants and animals from gold mining and related water issues. Retrieved Retrieved December 09, 2017 from www.ncbi.nlm.nih.gov/pubmed/15369321 Suspended Fundamentals Retrieved Solids of and Water Environmental December Clarity.” Measurements. 09, 2017 December 09, 2017 www.researchgate.net/publication/309513158_A_co mparative_study_on_lengthweight_relationship_len gthlength_relationship_and_condition_factor_of_a Fondriest Environmental, Inc. 2014. “Turbidity, Total the rivers Ganga,Yamuna and Gomti. Retrieved from Web.http://www.fondriest.com/environmental- _freshwater_snakehead_Channa_striata_from_the_ rivers_Ganga_Yamuna_and_Gomti Leopoldo A, Ihong M, Ihong M, Edorot R, Edorot J, Albutra Q. 2014. Describing Sexual Dimorphism in Mesopristes cancellatus in Tagoloan measurements/parameters/water-quality/turbidity- River Using Relative Warp Analysis. European total-suspended-solids-water-clarity. Journal of Zoological Research. 3, 15. Guidarelli G, Nicolosi P, Francesco M, Loy 2014. A. Fusco G, De Morphological variation and modularity in the mandible of three Mediterranean dolphin species, Italian Journal of www.researchgate.net/publication/306118130_Descr ibing_Sexual_Dimorphism_in_Mesopristes_cancella tus_in_Tagoloan_River_Using_Relative_Warp_Ana lysis Zoology 81(3), 354-367, Retrieved November 09, Macleod N. 2010. PalaeoMath: Part 21 - Principal 2017 from Warps, http://dx.doi.org/10.1080/11250003.2014.943685. Newsletter No.75. Retrieved November 13, 2017 from Relative Warps, and Procrustes PCA. www.palass.org/sites/default/files/media/palaeomat Hammer O. 2002. Morphometrics – brief notes. h_101/article_21/article_21.pdf Retrieved November 09, 2017 from http://folk.uio.no/ohammer/past/morphometry.pdf Medrano M, Jumawan J. 2016. Landmark-based geometric morphometric analysis of body shape Husin N. 2007 Morphological Genetic variation within population of Ibis fish, Ambassis Variability Of Malaysian Channa Spp Based On interrupta, collected from Masao River, Butuan City, Morphometric And Rapd Techniques. Retrieved Philippines Int’l Journal of Advances in Chemical November 13, 2017 from Engg., & Biological Sciences (IJACEBS) 3, Issue 2 http://eprints.usm.my/29331/1/Morphological_and (2016) ISSN 2349-1507 EISSN 2349-1515. Retrieved _genetic_variability_of_Malaysian_channa_spp_bas October 13, 2017 from. ed_on_morphometric_and_rapd_techniques.pdf http://iicbe.org/upload/4202AE0516113.pdf 191 Saura and Andante And Int. J. Biosci. 2018 Mineral Policy Center, 2000. Cyanide Leach Rohlf J. 2008. TpsRelw version 1.46 (Tps_Relative Mining Packet: Introduction. Online: February 19, Warps). Retrieved October 09, 2017 2017. http://life.bio.sunysb.edu/morph/ www.google.com.ph/url?sa=t&source=web&rct=j&ur l=https://www.earthworksaction.org/files/publicatio Rohlf J. 2008. TpsUtil version 1.40 (Tps_Utility). ns/Cyanide_Leach_Packet.pdf Retrieved October 09, 2017 http://life.bio.sunysb.edu/morph/ Ngilangil M, Boyles L, Sularte R. 2014. Abundance, Distribution and Conservation Status of Schutz H. 2007. Guide to Geometric Morphometrics Reptiles in Agusan Marsh, Bunawan, Agusan Del Sur, , version 0.4. Retrieved November 11, 2017 From Philippines: Retrieved November 14, 2017 from www.morphotools.net/softwareguide/GM%20guide http://dx.doi.org/10.15242/IJACEBS.C914015. %20v4%20LMs.pdf http://iicbe.org/upload/2528C91.4015.pdf Slice D, Bookstein F, Marcus L, Rohlf J, 2017. A Ortiz S. 2017. FISH BASE ORGANIZATION: List of Glossary for Geometric Morphometrics Updated Freshwater copy. Fishes reported from Philippines: Retrieved November 13, 2017 from Modified version 2017. Retrieved November 13, 2017 http://life.bio.sunysb.edu/morph/glossary/gloss1.ht from ml http://fishbase.org/country/CountryChecklist.php?s howAll=yes&what=list&trpp=50&c_code=608&cpres ence=Reported&sortby=alpha2&ext_CL=on&ext_pic =on&vhabitat=fresh Tah L, Gouli G, Da Costa K. 2012. Length-weight relationships for 36 freshwater fish species from two tropical reservoirs: Ayamé I and Buyo, Côte d’Ivoire Retrieved December 09, 2017. Park P, Aguirre W, Spikes D, Miyazaki J. 2013. www.scielo.sa.cr/scielo.php?script=sci_arttext&pid= Landmark-Based Geometric Morphometrics: What S0034-77442012000400036 Fish Shapes Can Tell Us about Fish Evolution. Tested Studies for Laboratory Teaching Proceedings of the Association for Biology Laboratory Education 34, 361-371. Retrieved October 13, 2017 from http://condor.depaul.edu/waguirre/park_et_al_13.p df Tubb R. 1977. Environmental Protection Agency, Office of Research and Development, Corvallis Environmental Research Laboratory, 1977 – 204. Retrieved December 09, 2017 from. https://books.google.com.ph/books?id=QCxGUyyI8 AUC&pg=PA160&lpg=PA160&dq=cyanide+affects+g Parker G. 1992. The Evolution Of Sexual Size rowth+fish&source=bl&ots=Z_2jaL_Xz7&sig=q_ISs Dimorphism In Fish. Journal of Fish Biology, 41: 1– SaoyQPESyYidlSfRAvrca0&hl=fil&sa=X&ved=0ahUK 20. Retrieved December 09, 2017, EwjmtJuktvvXAhULxrwKHUglBCcQ6AEISDAG#v=o http://dx.doi.org/10.1111/j.10958649.1992.Tb0386 nepage&q=cyanide%20affects%20growth%20fish&f= 4.X false Ramzy E. 2014. Toxicity and stability of sodium United States cyanide in fresh water fish Nile tilapia. Retreived Agency. 2017. September 08, 2017 from Quality Criteria. Online: March 16, 2017. http://dx.doi.org/110492914000198 Environmental National Protection Recommended State. or. WAIHI GOLD. 2015. Retrieved December Rohlf J. 2008. TpsDig version 2.12 (Tps_Digitize). 09, 2017 from Retrieved www.google.com.ph/url?q=http://arcweb.sos. October 09, http://life.bio.sunysb.edu/morph/. 2017 from www.waihigold.co.nz/mining/cyanide-ph-and-aciddrainage/cyanide/ 192 Saura and Andante Water Int. J. Biosci. Webster M, Introduction Morphometrics. 2018 Sheets to D. 2010. Landmark-Based in Quantitative A Practical Geometric Methods in Paleobiology, 163-188 P Paleontological Society Short Course, 193 Saura and Andante View publication stats Williams D. 2017. How Does Water Pollution Affect Fish? September 08, 2017 from https://sciencing.com/water-pollution-affect-fish4565696.