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      Cardiovascular Performance Measurement in Water Fleas by Utilizing High-Speed Videography and ImageJ Software and Its Application for Pesticide Toxicity Assessment

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          Abstract

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          With the advantages of easy culture, body transparency, and high sensitivity to chemical pollution, water fleas have been recognized as a good model for ecotoxicity studies. In this paper, we established ImageJ-based methods to measure cardiovascular performance by evaluating the heart rate and blood flow velocity in three water fleas for the first time. Among the three water fleas, Daphnia magna was identified as having the most robust heartbeat and blood flow rate and is therefore suitable for ecotoxicity assessment. Many important parameters like heart rate, blood flow rate, stroke volume, ejection fraction, fractional shortening, cardiac output heartbeat regularity can be extracted from videotaping and mathematical calculation. In utilizing those physiological parameters, the potential impacts of ambient water temperature and pesticide pollution on water fleas can be precisely measured.

          Abstract

          Water fleas are a good model for ecotoxicity studies, and were proposed for this purpose by the United States Environmental Protection Agency, due to their easy culture, body transparency, and high sensitivity to chemical pollution. Cardiovascular function parameters are usually used as an indicator of toxicity evaluation. However, due to the nature of the heart and blood flow, and the speed of the heartbeat, it is difficult to perform precise heartbeat and blood flow measurements with a low level of bias. In addition, the other cardiovascular parameters, including stroke volume, cardiac output, fractional shortening, and ejection fraction, have seldom been carefully addressed in previous studies. In this paper, high-speed videography and ImageJ-based methods were adopted to analyze cardiovascular function in water fleas. The heartbeat and blood flow for three water flea species, Daphnia magna, Daphnia silimis, and Moina sp., were captured by high-speed videography and analyzed using open-source ImageJ software. We found the heartbeat is species-dependent but not size-dependent in water fleas. Among the three water fleas tested, D. magna was identified as having the most robust heartbeat and blood flow rate, and is therefore suitable for the ecotoxicity test. Moreover, by calculating the diameter of the heart, we succeeded in measuring other cardiovascular parameters. D. magna were challenged with temperature changes and a pesticide (imidacloprid) to analyze variations in its cardiovascular function. We found that the heartbeat of D. magna was temperature-dependent, since the heartbeat was increasing with temperature. A similar result was shown in the cardiac output parameter. We also observed that the heartbeat, cardiac output, and heartbeat regularity are significantly reduced when exposed to imidacloprid at a low dose of 1 ppb (parts per billion). The blood flow rate, stroke volume, ejection fraction, and fractional shortening, on the contrary, did not display significant changes. In conclusion, in this study, we report a simple, highly accurate, and cost-effective method to perform physiological and toxicological assessments in water fleas.

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          Most cited references44

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          Reference right ventricular systolic and diastolic function normalized to age, gender and body surface area from steady-state free precession cardiovascular magnetic resonance.

          Recent advances in cardiovascular magnetic resonance (CMR) include improved image quality with steady-state free precession (SSFP) sequences and advanced post-processing of high temporal resolution ventricular function. We used these techniques to establish the reference values for right ventricular (RV) volumes and function. We studied 120 healthy subjects (60 men, 60 women; from 20 to 80 years) after exclusion of cardiovascular abnormality. Data were generated from SSFP cines, with three-dimensional modelling. Gender, body surface area (BSA), and age were independent predictors of several RV parameters. Normalized RV mass (RVM) and absolute and normalized RV volumes decreased significantly with age, whereas ejection fraction increased. For diastolic variables, absolute and normalized early peak filling rate (PFR(E)) decreased and absolute and normalized active peak filling rate (PFR(A)) in males increased, with decreased PFR(E)/PFR(A). Increasing BSA was associated with increased RVM, volumes, and PFR(E). Gender significantly influenced absolute and normalized mass and volumes, and absolute and normalized PFR(A). These data using state-of-the-art CMR show that normal values of RV systolic and diastolic parameters vary significantly by gender, BSA, and age. Appropriate reference ranges normalized to all three variables should be used in the determination of normality or severity of abnormality of RV dimensions and function.
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            Hazard identification of imidacloprid to aquatic environment.

            The use of a very effective insecticide against sucking pests, neonicotinoid imidacloprid, has been increasing extensively. For this reason elevated concentrations are expected in aquatic environment. Despite this fact, there is still a lack of data available on its possible risk for the environment. In this study, the potential hazards of imidacloprid and its commercial product Confidor SL 200 to aquatic environment were identified by the acute and chronic toxicity assessment using bacteria Vibrio fischeri, algae Desmodesmus subspicatus, crustacean Daphnia magna, fish Danio rerio and the ready biodegradability determination. We found out, that imidacloprid was not highly toxic to tested organisms in comparison to some other environmental pollutants tested in the same experimental set-up. Among the organisms tested, water flea D. magna proved to be the most sensitive species after a short-term (48 h EC50=56.6 mg L(-1)) and long-term exposure (21 d NOEC=1.25 mg L(-1)). On the contrary, the intensified toxicity of Confidor SL 200 in comparison to analytical grade imidacloprid was observed in the case of algae and slight increase of its toxicity was detected testing daphnids and fish. The activities of cholinesterase, catalase and glutathione S-transferase of daphnids were not early biomarkers of exposure to imidacloprid and its commercial product. Imidacloprid was found persistent in water samples and not readily biodegradable in aquatic environment. Due to increased future predicted use of commercial products containing imidacloprid and the findings of this work, we recommend additional toxicity and biodegradability studies of other commercial products with imidacloprid as an active constituent.
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              Left ventricular global systolic function assessment by echocardiography

              The left ventricle, with its thickened myocardial walls, unlike the right ventricle has no measurable geometric shape. It has a conical apex and its function quantification, needs intensive, 2D, 3D and M mode transesophageal echocardiography, which is described in this review.
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                Author and article information

                Journal
                Animals (Basel)
                Animals (Basel)
                animals
                Animals : an Open Access Journal from MDPI
                MDPI
                2076-2615
                05 September 2020
                September 2020
                : 10
                : 9
                : 1587
                Affiliations
                [1 ]Department of Bioscience Technology, Chung Yuan Christian University, Chung-Li 320314, Taiwan; fiorency_santoso@ 123456yahoo.co.id (F.S.); ferrysaputratj@ 123456gmail.com (F.S.)
                [2 ]Master Program of Nanotechnology, Chung Yuan Christian University, Chung-Li 320314, Taiwan
                [3 ]Papanin Institute for Biology of Inland Waters, Russian Academy of Sciences, Borok, Nekouzskii raion, Yaroslavl oblast 152742, Russia; krylovviacheslav@ 123456mail.ru
                [4 ]Faculty of Pharmacy, The Graduate School and Research Center for the Natural and Applied Sciences, University of Santo Tomas, Manila 1008, Philippines; alcastillo@ 123456ust.edu.ph
                [5 ]Department of Aquatic Biosciences, National Chiayi University, 300 Syuefu Rd. Chiayi 60004, Taiwan; s1070474@ 123456mail.ncyu.edu.tw
                [6 ]Department of Chemistry, Chung Yuan Christian University, Chung-Li 320314, Taiwan
                [7 ]Center for Nanotechnology, Chung Yuan Christian University, Chung-Li 320314, Taiwan
                Author notes
                [* ]Correspondence: hongthih@ 123456gmail.com (H.-T.L.); cdhsiao@ 123456cycu.edu.tw (C.-D.H.)
                Author information
                https://orcid.org/0000-0002-6167-3431
                https://orcid.org/0000-0002-6398-8672
                Article
                animals-10-01587
                10.3390/ani10091587
                7552287
                533d7862-0463-4bd2-9957-cb87d8ae3be5
                © 2020 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                : 04 August 2020
                : 04 September 2020
                Categories
                Article

                water flea,daphnia,cardiovascular performance,imagej
                water flea, daphnia, cardiovascular performance, imagej

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