1 National University of Saint Augustine (UNSA)Laboratory of Aquatic Biology, Academic Department of Biology, Arequipa, Peruvian HYDRA VULGARIS PALLAS, 1766 (HYDROZOA: HYDRIDAE) AS BIOINDICATOR OF THE WATER QUALITY OF THE RIVER CHILI, AREQUIPA, PERU Ronald Huarachi Olivera; Rosaura Gonzalez Juarez
2 I. Introduction tentacles testicle egg ovaryHydra vulgaris Pallas ,1766
3 I. Introduction Hydra vulgaris Pallas, 1766 (Hydrozoa: Hydridae) “freshwater Hydra” Test organism Easy breeding in laboratory Rapid reproduction Primary structure (Mesoglea, ectoderm, endoderm) Potential for location of chemical compounds Morphological changes under growing conditions of toxicity
4 c) Early stage short tentacles d) Short tentacles late phase b) Tentacles with bulbs REVERSIBLE SUBLETHAL EFFECTS a) Normal e) Tulip f) disintegrated EFECTOS IRREVERSIBLES LETALES
5 I. Introduction Freshwater Hydra test América ÁfricaPOTENTIAL TOXIC WATER SAMPLES Asia Europe LC50 (Median Lethal Concentration) CE50 (Median Effective Concentration)
6 I. Introduction The Chili River basin Southwest PeruChilean-Peruvian Coastal Desert Peaks in the cordillera occidental Mouth at Quilca in the Pacific Ocean
7 II. Material and MethodsCollection of Hydra vulgaris Springs “Ojo del Milagro” Locality of Cerrillo, Characato district to the southeast of the province of Arequipa. T° m= 17,8°C January2011 10 Km from the center of the city. 71º27´43,3´´LO, 16º28´16,6´´ LS; 2550msnm). = 57,22 mm Geophysical Institute UNSA T° m= 17,9°C February 2011 = 174,21 mm
8 II. Material and MethodsCultivation and maintenance of H. vulgaris Aquatic Biology Laboratory (UNSA) Table 1. Reagents used in the preparation of the Hydra culture medium (Castillo 2004). CaCl2-H2O ,94 g N-tris (hydroximetil) metil 1-2 aminoetanosulfónico, Buffer TES 2,2 g Acid etilén-diamino - tetraacético (EDTA) ,080 g distilled water L pH= 7,0 + 0,1 T°= °C
9 II. Material and MethodsDistilled water with activated carbon filtration Preparation of the culture medium Culture of Hydra vulgaris Photoperiod 12 h. light / h. darkness luminous Intensity =800 lux pH=7,0+0,1 T°=20+2°C
10 II. Material and MethodsEcotoxicological bioassays Filter Paper Water sample Sample problem (100 mL) 29,4 mg CaCl2 H2O 22 mg N-tris Bridge Tingo (71°33´49´´ LO; 16°25´47´´LS; 2233 msnm) 17 to 21 January 2011 Bridge Tiabaya (71°35´39´´ LO; 16°27´25´´ LS; 2134 msnm) 31 January to February 2011
11 II. Material and MethodsToxic Reference Sample problem K2Cr2O7 Bridge Tingo Bridge Tiabaya Concentrations (mg/L) Concentrations (%) CN 0,156 0,313 0,625 1,25 2,5 5,0 10,0 CN 1,56 3,13 6,25 12,5 25 50 100
12 II. Material and MethodsProbit method Median Lethal Concentration (LC50) Chi-cuadrado Morphological changes of Hydras 24 h. 48 h. 72 h. 96 h. Table 2. Toxicological categories of aqueous samples according to results obtained using bioassays ecotoxicological H. vulgaris (Castro et al. 2002). LC50 % Toxicity category Category Units of Toxicity (UT) < Very Toxic > 4,00 Toxic ,0 – 3,99 51 – Moderately toxic ,33 – 1,99 76 – Slightly toxic ,01 – 1,32 > Non-toxic < 1,00
13 II. Material and MethodsMethodology of the physical-chemical analysis of the water samples from the river Chili. Water sampling (15 cm) 1.- El pH pH-meter Digital Pocket (Hanna). 2.- Temperature Digital thermometer 3.- Nitrogen ammoniacal Colorimetry method. 4.- Phosphates Colorimetry method. 5.- Dissolved oxygen Method of Winkler. 6.- Electric conductivity Conductímetro 7.- Total Dissolved Solids portátil (Hanna)
14 II. Material y métodos 8.-DBO5 (5ºC) Laboratory Servilab (20ºC). 9.-Chrome Hexavalent (method 8023: 1,5 Di Phenylcarbohydrazide) 10.-Turbidity Disco Secchi Turbidity, Tº, pH, Dissolved oxygen, N-NH4 y Phosphates 3 Zonas Haven 3 Areas Table Fast
15 Bioassay Tingo negative control to different days of exposure, replica 11er day º day 3er day º day
16 Bioassay Tiabaya positive control at different days of exposure, replica 31er day º day 3er day º day
17 III. Results and DiscussionUnits of toxicity (UT) 100 LC50% = Table 3. Mortality of H. vulgaris and Lethal Concentration Media (LC50%) due to the collected waters of Tingo River Chili - Arequipa, Peru. Concentrations (%) Time(h) LC50(%) , , , , Mortality (%) > > > ,
18 III. Results and Discussiona) 72 hours b) 96 hours Percentage of mortality and LC50 of the collected waters of the river Chili at the point of sampling of Tingo on Hydra vulgaris at the 72 to 96 hours.
19 III. Results y DiscussionTable 4. Mortality of H. vulgaris and lethal Media concentration (LC50%) due to the collected waters of Tiabaya in Chili River. Arequipa, Peru. Concentrations (%) Time LC , , , , (h) (%) Mortality (%) , , , ,
20 III. Results y Discussiona) 24 hours b) 48 hours c) 72 hours d) 96 hours Percentage of mortality and LC50 of the collected waters of the river Chili at the point of sampling of Tiabaya on H. vulgaris at 24, 48, 72 and 96 hours..
21 Table 5. Registration percentage of morphological changes of HTable 5. Registration percentage of morphological changes of H. vulgaris due to exposure to different concentrations and exposure to potassium bichromate times K2Cr2O7.
22 Table 6. Registration percentage of morphological changes of HTable 6. Registration percentage of morphological changes of H. vulgaris due to exposure to different concentrations and exposure times in bioassays with waters of Tingo, Arequipa, Peru.
23 Table 7. Registration percentage of morphological changes of HTable 7. Registration percentage of morphological changes of H. vulgaris due to exposure to different concentrations and exposure time in bioassays with waters of Tiabaya - Arequipa, Peru.
24 III. Results and DiscussionTable 8. Lethal Concentration Media (LC50%) , toxicological categories and units of toxicity (UT) concentration for the results of the bioassay with H. vulgaris at different exposure times depending on the two points of sampling of water. Sampling points Time LC50 % Toxicologicas Units of Toxicity (h) Categories (UT) Bridge Tingo , ,74 Bridge Tiabaya , ,62 Bridge Tiabaya , ,26 Bridge Tiabaya , ,61 Bridge Tiabaya , ,66 1= Very Toxic. 2 = Toxic =Moderately Toxic = Slightly Toxic. 5 = Non-Toxic
25 III. Results and DiscussionTable 9. Physical chemical parameters of the water of the two sampling points of water on Chili River, Arequipa, Peru 2011 and comparison with Peruvian environmental quality standards (ECAs). Categoría 3* Categoría 4* Parameter Analysis Unit Tingo Tiabaya Irrigation of Drink Of Conservation plants of low aquatic of the aquatic and high stem animals medium Transparency in situ cm , , Temperature in situ °C , , pH in situ , , ,5 -8, , ,5 OD Laboratory mg·L-1 5, , > > >5 N-NH4 Laboratory mg·L-1 0, , ,02 PO Laboratory mg·L-1 0, , ,5 DBO Laboratory mg·L -1 6, , < <10 CE Laboratory uS·cm < < TDS Laboratory ppm Cr Laboratory mg·L-1 0, , , ,05 *National Peruvian environmental quality (ECA) standards for water.
26 IV. Conclusions The results of the physicochemical River chili parameters, were compared with the Peruvian national standards of environmental quality (ACE) for water (categories 3 and 4) and high values of biochemical oxygen demand (BOD5) and NH4 (PO4) 3-, and low in dissolved oxygen (OD) for Tiabaya values were recorded. In Tingo, NH4 and phosphates were high. In Tiabaya, water was considered most toxic Tingo.
27 Collector Alata - Tiabaya
28 Collector Alata - Tiabaya
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31 V. ACKNOWLEDGEMENTS Cesar Bernabe Ortiz, Edwin Bocardo Delgado, Herbert Lazo Rodríguez, for the support they gave me in this research and friends of biology: Tito Cuadros, Norbert Tejada, Alex Dueñas, Marco Ríos, Juan Pablo Portilla, Raúl Lima, Edwin Bustamante, Erick Huamani, Yvonne Medina and Roberto Fernández for the help they gave me during the course of this work.
32 Thanks a lot