Bioindicator and biomarker for toxicity of pesticide

To measure the toxicity of pesticide influence the wildlife nearby, different organisms are needed in different situation.(1, 2) Different number of factors like gender, age and body weight  can affect the susceptibility of individual, thus population-level measure of toxicity are considered.(2, 3) There are different parameters in measure the toxicity of pesticide on the wildlife based on different situation like lipophilicity of the toxicant and expected toxicity on individual organisms.(4) Many pesticide may not show immediate effect in particular organisms, thus long term observe may need or proxy are needed for estimation in case of endangered species.(4)

LC50 as the most common test for acute toxicity tests
LC50 is measure of toxicity of toxicant with statistically derived dose that kill 50% of the sample population of a specific test-animal specified period through exposure.(5) LC50 is a comparative tool more than a predictive one, it enable the comparison of single species under different levels of same toxicant.(5) Different animal has different LC50 value on the same toxicant, such as human has higher LC50 value on theobromine when compare to dog because higher level of cytochrome P450 available for break down the compound.(6) LC50 in laboratory may has lot of differences when compare to the death of organism in habitat under pollution of chemical pesticide as soil conditions or water conditions can change the solubility of the chemical pesticide, the metabolite of chemical pesticide maybe more toxic than the original form after degraded in the soil and water etc.(7) Also, there may have some hidden variable on population decline of certain wildlife such as introduction of new invasive species in the same environment.(8)     

Different bioindicators used in different purpose.
For acute toxicity under aquatic environment, small organism such as Artemia salina can be used as they showed rapid response to toxicant.(9) For acute toxicity under soil environment, earthworm can be used to tested for toxicity after soil sprayed or mixed with chemical pesticide, death are counted on day 14 and day 56.(10) Other animal such as bees can be used for targeting certain species population decline under influence of pesticide.(11) Plants can be used for measuring strength of herbicide toxicity.(12) The ideal bioindicator should be abundant, easy to sample, territorial / sedentary, adequate size, well known biology, stability of populations and ubiquitous in space with time (13)

 

Different control group are used in different situation.
There are several control groups used during testing of pesticide toxicity which is negative control, positive control, carrier control and sham control.(14) Negative control is used when the toxicity of pesticide are not known, the control group are not administrate with toxicant.(15) Positive control is used when the toxicity of pesticide are known, the control group are administrated with toxicant and expected to have effect.(16) Carrier control are used when the pesticide are not lipophilic, the pesticide is dissolved in carrier such as corn oil, the control group are administrated with corn oil to ensure the vehicle doesn’t cause effect on it.(17) Sham control are used when physical actions like injections are performed, toxicologist goes through the motions like injection without actually given the toxicant to ensure the organism is reacted with toxicant rather than the physical action.(18)

Different biomarkers are selected for different pollutant.

Biomarkers are quantifiable biochemical, physiological, or histological changes in an organism, a strong footprint for finding contamination in the body.(19) It also help finding the cause-and-effect relationships between an exposure to contaminants and biological responses.(19, 20)

Biomarkers can be further divided into biomarker of exposure, biomarker of effect and biomarkers of susceptibility.(21-23) Biomarker of exposure indicated that organism are contaminated with pesticide but adverse effect are not related.(21) Biomarker of effect indicated that the organism is suffered from a known adverse effect.(22) Biomarker of susceptibility indicates there are natural characteristics that increases susceptibility to experience adverse effects like oncogene that able to activate the cancer process of the animal.(23)

 

Different pesticide has different chemical properties, so the organism has different biomarkers react to the toxicant.(24-29) The biomarkers mainly divided into metallic and organic pollutant.(24-29)

For metallic contaminant, the biomarkers are related to oxidative stress and metallic protein.(24-26) Metallothioneins are used as those metallic compounds tends to bind with this protein.(24) Oxidative stress are used as many metallic ion can disturb normal redox state in cellular environment, which cause toxic effects through the production of peroxides and free radicals that damage all components of the cell. (25) Stress Proteins (HSPs) are used as metallic contaminant cause stressful conditions in cellular environment, those protein are secreted in order to counter the damage done by metallic ion.(26)

For organic contaminant, the biomarkers are related to DNA mutation and lipid.(27-29) EROD activity are used as evidence of receptor-mediated induction of cytochrome P450-dependant monooxygenases, which indicate the activation of enzyme system to filter the toxin in the liver.(27) PAH biliary metabolites are used for evidence of polycyclic aromatic hydrocarbon present in bile secretion, which indicate there are organic pollutant dissolved in the lipid within the body.(28) DNA adducts are used as evidence for DNA damage in the cell, which indicate the organism are at risk for developing cancer under contamination of pesticide.(29)

 

Field monitoring as part of plan for chemical pesticide monitoring
Environment is constantly change, single species toxicity tests are inadequate for assessing ecological field impacts as they are co-varied together.(30, 31) Multiple floras and faunas gather as one ecosystem, which formed complicated biological interaction, thus cannot separate into individual components for investigate pesticide pollution.(30) Many pesticide contamination in aquatic environment are origin from diffuse source as those chemical are water soluble, therefore field monitoring help finding the cause and control the level of agricultural chemicals in the water.(31)    

  

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2.     Amado LL, Garcia ML, Ramos PB, Freitas RF, Zafalon B, Ferreira JLR, et al. A method to measure total antioxidant capacity against peroxyl radicals in aquatic organisms: Application to evaluate microcystins toxicity. Science of The Total Environment. 2009;407(6):2115-23.

3.     Costa LG, Giordano G, Cole TB, Marsillach J, Furlong CE. Paraoxonase 1 (PON1) as a genetic determinant of susceptibility to organophosphate toxicity. Toxicology. 2013;307:115-22.

4.     Sarin H. Prediction and characterization of toxic potential of occupational and environmental toxins & toxicants on the basis of conserved biophysical properties: Lipophilicity or hydrophilicity in context of molecular size and molecular charge [M.P.H.]. Ann Arbor: Icahn School of Medicine at Mount Sinai; 2014.

5.     Zolotarev KV, Nakhod KV, Mikhailov AN, Mikhailova MV. Correlation between LC50 for Adult Fish and Fish Embryos. Bulletin of Experimental Biology and Medicine. 2017;163(6):749-52.

6.     Gans JH, Korson R, Cater MR, Ackerly CC. Effects of short-term and long-term theobromine administration to male dogs. Toxicology and Applied Pharmacology. 1980;53(3):481-96.

7.     Mugni H, Paracampo A, Solis M, Fanelli S, Bonetto C. Acute toxicity of roundup to the nontarget organism Hyalella curvispina. Laboratory and field study. Toxicological & Environmental Chemistry. 2014;96(7):1054-63.

8.     Macek J, Sipek P. Azalea sawfly Nematus lipovskyi (Hymenoptera: Tenthredinidae), a new invasive species in Europe. European Journal of Entomology. 2015;112(1):180.

9.     Ozkan Y, Altinok I, Ilhan H, Sokmen M. Determination of TiO2 and AgTiO2 Nanoparticles in Artemia salina: Toxicity, Morphological Changes, Uptake and Depuration. Bulletin of Environmental Contamination and Toxicology. 2016;96(1):36-42.

10.   Selin-Rani S, Senthil-Nathan S, Thanigaivel A, Vasantha-Srinivasan P, Edwin E-S, Ponsankar A, et al. Toxicity and physiological effect of quercetin on generalist herbivore, Spodoptera litura Fab. and a non-target earthworm Eisenia fetida Savigny. Chemosphere. 2016;165:257-67.

11.   Decourtye A, Henry M, Desneux N. Overhaul pesticide testing on bees. Nature. 2013;497:188.

12.   Ray S, White W. Selected aquatic plants as indicator species for heavy metal pollution. Journal of Environmental Science and Health  Part A: Environmental Science and Engineering. 1976;11(12):717-25.

13.   Fossi MC, Sànchez-Hernàndez JC, Dìaz-Dìaz R, Lari L, Garcia-Hernàndez JE, Gaggi C. The lizard Gallotia galloti as a bioindicator of organophosphorus contamination in the Canary Islands. Environmental Pollution. 1995;87(3):289-94.

14.   Rohwer G. Treatment and Control Groups in a Dynamic Setting. International Statistical Review. 2016;84(1):63-78.

15.   Vieira Santos VS, Caixeta ES, Campos Júnior EOd, Pereira BB. Ecotoxicological effects of larvicide used in the control of Aedes aegypti on nontarget organisms: Redefining the use of pyriproxyfen. Journal of Toxicology and Environmental Health, Part A. 2017;80(3):155-60.

16.   Klimisch HJ, Andreae M, Tillmann U. A Systematic Approach for Evaluating the Quality of Experimental Toxicological and Ecotoxicological Data. Regulatory Toxicology and Pharmacology. 1997;25(1):1-5.

17.   Hallare A, Nagel K, Köhler H-R, Triebskorn R. Comparative embryotoxicity and proteotoxicity of three carrier solvents to zebrafish (Danio rerio) embryos. Ecotoxicology and Environmental Safety. 2006;63(3):378-88.

18.   Byzitter J, Lukowiak K, Karnik V, Dalesman S. Acute combined exposure to heavy metals (Zn, Cd) blocks memory formation in a freshwater snail. Ecotoxicology. 2012;21(3):860-8.

19.   Peakall DB. The role of biomarkers in environmental assessment (1). Introduction. Ecotoxicology. 1994;3(3):157-60.

20.   Forbes VE, Palmqvist A, Bach L. The use and misuse of biomarkers in ecotoxicology. Environmental Toxicology and Chemistry. 2006;25(1):272-80.

21.   Tyler CR, van der Eerden B, Jobling S, Panter G, Sumpter JP. Measurement of vitellogenin, a biomarker for exposure to oestrogenic chemicals, in a wide variety of cyprinid fish. Journal of Comparative Physiology B. 1996;166(7):418-26.

22.   Biggs ML, Kalman DA, Moore LE, Hopenhayn-Rich C, Smith MT, Smith AH. Relationship of urinary arsenic to intake estimates and a biomarker of effect, bladder cell micronuclei. Mutation Research/Reviews in Mutation Research. 1997;386(3):185-95.

23.   Costa LG, Cole TB, Vitalone A, Furlong CE. Measurement of paraoxonase (PON1) status as a potential biomarker of susceptibility to organophosphate toxicity. Clinica Chimica Acta. 2005;352(1):37-47.

24.   Cobbett C, Goldsbrough P. PHYTOCHELATINS AND METALLOTHIONEINS: Roles in Heavy Metal Detoxification and Homeostasis. Annual Review of Plant Biology. 2002;53(1):159-82.

25.   Sayeed I, Parvez S, Pandey S, Bin-Hafeez B, Haque R, Raisuddin S. Oxidative stress biomarkers of exposure to deltamethrin in freshwater fish, Channa punctatus Bloch. Ecotoxicology and Environmental Safety. 2003;56(2):295-301.

26.   Sanders BM. Stress Proteins in Aquatic Organisms: An Environmental Perspective. Critical Reviews in Toxicology. 1993;23(1):49-75.

27.   Gagnon MM, Rawson CA. Bioindicator species for EROD activity measurements: A review with Australian fish as a case study. Ecological Indicators. 2017;73:166-80.

28.   Vuorinen PJ, Keinänen M, Vuontisjärvi H, Baršienė J, Broeg K, Förlin L, et al. Use of biliary PAH metabolites as a biomarker of pollution in fish from the Baltic Sea. Marine Pollution Bulletin. 2006;53(8):479-87.

29.   Ching EWK, Siu WHL, Lam PKS, Xu L, Zhang Y, Richardson BJ, et al. DNA Adduct Formation and DNA Strand Breaks in Green-lipped Mussels (Perna viridis) Exposed to Benzo[a]pyrene: Dose- and Time-Dependent Relationships. Marine Pollution Bulletin. 2001;42(7):603-10.

30.   Webb TJ. Marine and terrestrial ecology: unifying concepts, revealing differences. Trends in Ecology & Evolution. 2012;27(10):535-41.

31.   Carter A. How pesticides get into water - and proposed reduction measures. Pesticide Outlook. 2000;11(4):149-56.

 

 

Figure 1: Artemia salina is one of the common bioindicator for ecotoxicology studies, it also used for children's toy

Figure 2: Bee is also one of the bioindicator for measuring pesticide toxicity