Next Article in Journal
Survey of Microcystin-Producing Cyanobacteria in French Lakes of Various Trophic Status Using Environmental and Cyanobacterial Parameters and an Active Mussel Biomonitoring
Previous Article in Journal
Dual Proteomics Strategies to Dissect and Quantify the Components of Nine Medically Important African Snake Venoms
 
 
Search for Articles:
Title / Keyword
Author / Affiliation / Email
Journal
Article Type
 
 
Advanced Search
 
Section
Special Issue
Volume
Issue
Number
Page
 
Journals
Toxins
Volume 17
Issue 5
10.3390/toxins17050244
toxins-logo
Submit to this Journal Review for this Journal Propose a Special Issue
► Article Menu

Article Menu

share Share announcement Help format_quote Cite question_answer Discuss in SciProfiles
first_page
settings
Order Article Reprints
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:
Background:
Article

Toxicity Assessment of Catechins on Representative Aquatic Organisms and Terrestrial Plant

by
Khawaja Muhammad Imran Bashir
1,2,†,
Hye-Ryeon An
1,†,
Bertoka Fajar Surya Perwira Negara
3,
Gabriel Tirtawijaya
4,
Maria Dyah Nur Meinita
5,6,
Jae-Hak Sohn
7,
Dicky Harwanto
8,* and
Jae-Suk Choi
1,*
1
Department of Seafood Science and Technology, The Institute of Marine Industry, Gyeongsang National University, Tongyeong 53064, Republic of Korea
2
German Engineering Research and Development Center, LSTME-Busan Branch, Busan 46742, Republic of Korea
3
Department of Marine Science, University of Bengkulu, Jl. W.R Soepratman, Bengkulu 38371, Indonesia
4
Faculty of Biotechnology, University of Surabaya, Jalan Raya Kalirungkut Surabaya, Surabaya 60292, Indonesia
5
Faculty of Fisheries and Marine Science, Jenderal Soedirman University, Purwokerto 53123, Indonesia
6
Center for Maritime Bioscience Studies, Jenderal Soedirman University, Purwokerto 53123, Indonesia
7
Department of Food Science and Culinary Arts, College of Health and Welfare, Silla University, Busan 46958, Republic of Korea
8
Faculty of Fisheries and Marine Science, Diponegoro University, Semarang 50275, Indonesia
*
Authors to whom correspondence should be addressed.
†
These authors contributed equally to this work.
Toxins 2025, 17(5), 244; https://doi.org/10.3390/toxins17050244
Submission received: 11 April 2025 / Revised: 5 May 2025 / Accepted: 12 May 2025 / Published: 14 May 2025
Browse Figure
Versions Notes

Abstract

:
Catechins, renowned for their health benefits, have unexamined environmental impacts. This study assessed the toxicity of crude catechin and catechin hydrate on invertebrate larvae, plant, and microalgae. The survival rates of Daphnia magna Straus and Artemia salina L. were monitored every 24 h over a three-day period. The germination rate and radicle length of Lactuca sativa L. was measured every 24 h for four days. Inhibitory effects were evaluated in both freshwater and seawater cultures of Chlorella vulgaris Beijerinck, with cell density recorded every 24 h and yield inhibition calculated after 96 h. Results indicated that increasing catechin concentration and exposure duration decreased the survival rate of D. magna and A. salina. Daphnia magna was more sensitive to catechins than A. salina, with 24 h lethal concentration 50 (LC-50) values of 1174 µg/mL compared to 1895 µg/mL for crude catechin, and 54 µg/mL compared to 153 µg/mL for catechin hydrate. The germination rate and radicle length of L. sativa, along with the cell density of C. vulgaris, decreased with increasing catechin concentration, but remained higher even after prolonged exposure. At low catechin concentrations, C. vulgaris cell density exceeded control levels. This study demonstrates that catechins in aquatic environments can significantly impact ecosystems. At certain concentrations, catechins are toxic and potentially lethal to aquatic organisms. Conversely, at lower concentrations, catechins may promote microalgal growth, suggesting a fertilizing effect. Understanding these dynamics is crucial for maintaining the stability of aquatic ecosystems.
Keywords:
algae; cell density; eco-friendly; germination; plankton; toxicity
Key Contribution: The manuscript highlights the minimal environmental toxicity of catechins, specifically crude catechin and catechin hydrate, while demonstrating their potential to enhance growth at certain concentrations. This study provides valuable insights into the environmental applications of catechins, suggesting their dual role as safe substances and potential growth enhancers for plants and microalgae.

1. Introduction

Catechins are a type of flavonoid, which are plant polyphenols that include epicatechin (EC), epigallocatechin (EGC), epicatechin gallate (ECG), and epigallocatechin gallate (EGCG) as diastereoisomers [1]. Among these, catechin hydrate is a natural flavonoid known for its antioxidant and anti-inflammatory properties. It is beneficial not only for prevention but also for the treatment of diseases caused by oxidative damage, such as inhibiting cancer growth [2,3]. As important dietary components, catechins are found in plants like green tea, nuts, chocolate, red wine, and fruits such as apricots, black grapes, cocoa, and cherries [4,5]. These compounds serve as antioxidants, antimicrobials, and anticarcinogens [6,7]. Their efficiency as antioxidants surpasses many other substances, making them a cost-effective choice in food production [8]. Studies on both animals and humans have demonstrated that catechins impact lipid metabolism by reducing triglycerides, total cholesterol [9], and body fat accumulation [10,11,12]. By binding strongly to reactive oxygen species (ROS), they absorb free radicals [1]. As environmentally friendly plant polyphenols [13], even minimal doses can lead to health improvements [14].
The toxic effects of catechins on aquatic environments need to be evaluated. Freshwater and saltwater ecosystems have distinct characteristics that influence the sensitivity of their resident biota. Studies on the effects of catechins on both freshwater and marine organisms are crucial in understanding the broader ecological risks. In the present study, we utilized several organisms: Daphnia magna Straus neonates, Artemia salina L. nauplii, Lactuca sativa L. seeds, and Chlorella vulgaris Beijerinck. D. magna is a commonly used freshwater organism in environmental studies [15,16], particularly for assessing drug toxicity [17,18,19,20]. It is chosen for toxicity measurement due to its sensitive behavioral and physiological responses to environmental factors and chemicals, making it a biomarker in studies on phenotype, toxicology, and life cycle evolution [21]. Daphnia magna is a popular freshwater organism for bioassays in biological and environmental studies, with A. salina serving as the seawater equivalent. This species is used in food safety [22], medical safety [23], therapeutic and pharmacological [24,25,26], and eco-toxicity studies [15,16,27,28].
Lactuca sativa is a macro plant frequently used in bioassays and environmental quality bioindicator studies due to its quick and easy germination [29]. Its sensitivity to environmental changes makes it suitable for water [30] and soil quality studies [22], as well as studies on heavy metal extraction [31] and air pollution [32]. Lactuca sativa has also been used in bioherbicide [33] and allelochemical studies [34]. Microalgae, another aquatic organism, is useful for monitoring environmental pollution and conducting aquatic toxicity tests [16,35]. Chlorella vulgaris is economically and effectively used in toxicity research due to its sensitivity to toxicants and ease of laboratory cultivation [16].
Previous studies have indicated that catechins are more toxic to Gram-positive bacteria (Bacillus subtilis) than to Gram-negative bacteria (Escherichia coli) [1]. Additionally, high doses of catechins can cause liver side effects, though this is not the case when consumed as brewed tea, as beverage extracts, or in food form [36]. The appropriate concentration of catechins has been shown to reduce mortality in brine shrimp when exposed to toxic methanol concentrations [37].
Daphnia magna and A. salina are crucial components of aquatic ecosystems. Daphnia magna, found in freshwater, feeds on algae and phytoplankton, helping to control algal blooms (eutrophication) and serving as a vital food source for fish and invertebrates. Artemia salina, which inhabits brackish water and seawater, consumes microorganisms and detritus, contributing to ecosystem balance. It also supports the carbon and nitrogen cycles and is an important food source for fish and invertebrates. Both species play key roles in the aquatic food web and help maintain ecological stability.
Lactuca sativa, although not an aquatic macrophyte, shows potential for use in aquatic systems through its phytoremediation abilities. In terrestrial–aquatic integrated systems, it can absorb inorganic nitrogen compounds (e.g., nitrates) and other pollutants from aquatic organism waste, helping stabilize water chemistry and improve system efficiency. Chlorella vulgaris, a single-celled microalga, is a primary producer in aquatic environments. Through photosynthesis, it generates oxygen and organic matter that support the food chain. It also absorbs excess nutrients like nitrogen and phosphate, reducing eutrophication, and can capture heavy metals and toxins, making it effective for bioremediation in polluted waters.
By conducting this study, we aim to generate basic knowledge about the significant impacts of catechins on the stability of aquatic ecosystems. Understanding these impacts is essential, as catechins, whether naturally occurring or arising through environmental exposure, can potentially disrupt ecological balance, affect species survival, and alter water quality. This study is expected to contribute to environmental risk assessment and inform future strategies for managing the presence of these compounds in aquatic environments.

2. Results

2.1. HPLC Analysis of Crude Catechin and Catechin Hydrate

For the high-performance liquid chromatography (HPLC) analysis, catechin hydrate standard (50 µg/mL), crude catechin (200 µg/mL), and a combination of both were injected into the HPLC instrument. The resulting chromatogram is shown in Figure 1. Catechin retention times were observed to range from 3.0 to 3.5 min.
Catechin was successfully identified in the extract samples. In the chromatogram of the pure catechin hydrate standard (Figure 1a), a distinct peak was observed at a retention time characteristic of catechin. The extract sample (Figure 1b) exhibited a peak at the same retention time, indicating the presence of catechin. To further confirm this identification, a co-injection of the extract and catechin standard was performed (Figure 1c). The resulting chromatogram displayed a single intensified peak at the same retention time, with no peak splitting or additional signals. This co-elution demonstrated that the compound in the extract matched the standard completely. This confirms the identity of the compound as catechin, showing qualitative agreement and chromatographic consistency, and providing strong evidence for its presence in the extract.

2.2. Larvicidal Effects of Crude Catechin and Catechin Hydrate on the Survival of D. magna

The larvicidal effects on D. magna neonates are detailed in Table 1. The control group of neonates exhibited high survival rates of 100%, 94%, and 85% at 24, 48, and 72 h, respectively.
Crude catechin showed a larvicidal effect at concentrations of 2.0 and 3.0 mg/mL after 24 h, with the most significant effect observed at 3.0 mg/mL, resulting in only 10% survival. The lethal concentration 50 (LC-50), which is the concentration required to kill 50% of the organisms, was determined to be 1174 µg/mL at 24 h. At a concentration of 0.5 mg/mL, crude catechins showed a mild larvicidal effect, with survival rates of 43% at 48 h and 28% at 72 h. At 48 and 72 h, no neonates survived at concentrations of 2.0 or 3.0 mg/mL.
Catechin hydrate demonstrated the most severe larvicidal effect at a concentration of 0.5 mg/mL after 24 h, with 0% survival. At a concentration of 0.1 mg/mL for 24 h, the survival rate was 38%. The LC-50 at 24 h was found to be 54 µg/mL. After 72 h, all tested concentrations showed a larvicidal effect. Specifically, a concentration of 0.01 mg/mL resulted in a 40% survival rate, while 0.05 mg/mL resulted in a 20% survival rate. The LC-50 at 72 h was determined to be 7 µg/mL.

2.3. Larvicidal Effects of Crude Catechin and Catechin Hydrate on the Survival of A. salina

In this study, pure seawater was used as a negative control and did not kill A. salina after 24 h, with minimal mortality observed at 48 h (2%) and 72 h (7%). The larvicidal effects of crude catechin on A. salina nauplii are detailed in Table 2. After 24 h, a concentration of 0.5 mg/mL resulted in 100% survival, but higher concentrations induced mortality, with 3.0 mg/mL causing the highest mortality rate of 84%. The LC-50 at 24 h was determined to be 1895 µg/mL. At 48 h, concentrations of 2.0 and 3.0 mg/mL resulted in no surviving A. salina nauplii, while a concentration of 1.0 mg/mL had a low survival rate of 20%.
Catechin hydrate showed larvicidal effects at concentrations of 0.1 and 0.5 mg/mL after 24 h, with survival rates of 45% and 40%, respectively. The LC-50 at 24 h was 153 µg/mL. After 72 h, A. salina nauplii exposed to concentrations of 0.01 and 0.05 mg/mL showed survival rates of 42% and 23%, respectively. The LC-50 at 72 h was determined to be 9 µg/mL.

2.4. Inhibitory Effect of Crude Catechin and Catechin Hydrate on Germination of L. sativa

The toxicity and inhibition levels of crude catechin and catechin hydrate on the germination and radical growth of L. sativa are summarized in Table 3. Lactuca sativa seeds treated with 1.0 mg/mL crude catechin exhibited a higher germination rate of 87.5% compared to the control at 24 h. However, at a concentration of 50.0 mg/mL, germination did not occur. At 1.0 mg/mL, radicle growth was slower than that of the control, with lengths of 3.11 mm compared to 3.84 mm for the control. At 10.0 mg/mL, germination was 35% at 24 h and 75% at 96 h, both lower than that of the control.
Catechin hydrate resulted in longer radicle growth at concentrations of 0.1, 0.5, and 1.0 mg/mL. At 0.1 mg/mL, radical length was 4.47 mm at 24 h, compared to 4.10 mm for the control. At 5.0 mg/mL, radical length was 2.95 mm. After 48 h, a concentration of 0.1 mg/mL catechin hydrate resulted in a radical length of 13.26 mm, compared to 15.74 mm for the control. Germination rates were lower for catechin hydrate, with an 83.3% germination rate at 0.1 mg/mL compared to 93.3% for the control at 96 h.

2.5. Toxicity of Crude Catechin and Catechin Hydrate on Cell Density of Freshwater C. vulgaris

The effects of crude catechin and catechin hydrate on the cell density of freshwater C. vulgaris are detailed in Table 4. After 24 h, crude catechin at concentrations of 0.5 and 1.0 mg/mL significantly increased cell density, with values of 1.50 × 106 cells/mL and 1.42 × 106 cells/mL, respectively, compared to the control at 1.16 × 106 cells/mL. However, a concentration of 2 mg/mL showed no significant growth enhancement. At 72 h, a concentration of 1.0 mg/mL resulted in a lower cell density than that of the control, while 0.5 mg/mL showed no significant difference. Yield inhibitions for catechin hydrate were 12.29 ± 1.60% and 33.84 ± 0.60% at concentrations of 1.0 and 2.0 mg/mL, respectively, with lower concentrations showing approximately a four-fold increase in yield.
Catechin hydrate increased cell density at all concentrations after 24 h, with values ranging from 1.33 × 106 to 1.59 × 106 cells/mL. From 48 h to 96 h, a concentration of 1.0 mg/mL did not exceed the cell density of the control. From 72 h to 96 h, a concentration of 0.5 mg/mL did not exceed the control cell density, but lower concentrations resulted in increased cell density.

2.6. Toxicity of Crude Catechin and Catechin Hydrate on Cell Density of Seawater C. vulgaris

The effects of crude catechin on the cell density of seawater C. vulgaris are presented in Table 5. After 24 h, there were no significant changes in cell density compared to the control. However, significant changes were observed at 48 h, with concentrations of 0.5 and 2.0 mg/mL resulting in cell densities of 1.90 × 106 and 1.37 × 106 cells/mL, respectively. By 96 h, all seawater C. vulgaris treatments showed significantly different cell density values. Concentrations of 1.0 and 2.0 mg/mL resulted in lower cell density values (2.63 × 106 and 1.61 × 106 cells/mL, respectively) compared to that of the control (3.96 × 106 cells/mL). At 0.5 mg/mL, cell density increased by 2.61%, while concentrations of 1.0 and 2.0 mg/mL led to inhibitions of 19.90% and 35.26%, respectively.
When exposed to catechin hydrate, seawater C. vulgaris showed significant changes in cell density compared to the control after 24 h at all concentrations. The lowest concentration resulted in a cell density of 1.36 × 106 mg/mL, while at 0.1 mg/mL, cell density was 1.56 × 106 mg/mL. Higher concentrations (0.5 and 1.0 mg/mL) decreased cell density to 1.35 × 106 and 1.32 × 106 cells/mL, respectively. After 48 h, concentrations of 0.05 and 0.1 mg/mL showed no significant differences in cell density (2.21 × 106 and 2.20 × 106 cells/mL) compared to the control (2.29 × 106 mg/mL). After 96 h, all concentrations except 0.1 mg/mL were significantly different from that of the control. Yield inhibitions for concentrations of 0.5 and 1.0 mg/mL were 16.37 ± 1.53% and 35.97 ± 1.70%, respectively, while lower concentrations (0.05 and 0.1 mg/mL) increased cell density by 5.92% and 1.71%, respectively.

3. Discussion

In this study, D. magna was placed on 24-well plates with 2 mL of solution, minimizing the use of compounds and bioactive reagents. Crude catechins showed mild larvicidal effects at 2.0 mg/mL and high larvicidal effects at 3.0 mg/mL after 24 h, with survival rates of 45.0 ± 4.1% and 10.0 ± 4.1%, respectively (LC-50 at 24 h: 1174 µg/mL). At 48 h, 0.5 mg/mL showed mild effects, while 1.0 mg/mL showed high effects, and 2.0 and 3.0 mg/mL resulted in 100% mortality. Catechin hydrate at 0.5 mg/mL resulted in 0% survival at 24 h (LC-50 at 24 h: 54 µg/mL). At 0.1 mg/mL, it had mild larvicidal effects, with a survival rate of 38.0 ± 2.9%. The increase in mortality is likely due to the higher concentrations and longer exposure times [38]. Catechin hydrate was more toxic to D. magna than crude catechin, consistent with Hu et al. [36], who found fewer side effects from catechins in brewed tea or food than in pure form.
For A. salina nauplii, crude catechins at 0.5, 1.0, and 2.0 mg/mL did not show larvicidal effects up to 24 h, while 3.0 mg/mL did show effects (LC-50 at 24 h: 1895 µg/mL). At 48 h, 2.0 and 3.0 mg/mL resulted in 100% mortality, with 1.0 mg/mL showing mild effects (20.0 ± 4.1% survival rate). Catechin hydrate was more toxic to A. salina than crude catechin, with mild effects at 0.1 and 0.5 mg/mL after 24 h (survival rates of 45.0 ± 5.0% and 40.0 ± 5.0%, respectively). The LC-50 at 24 h was observed to be 153 µg/mL. At 48 h, 0.01 and 0.05 mg/mL showed no larvicidal effects, with a survival rate of 80% and 52%, respectively. The LC-50 at 48 h was 70 µg/mL. After 72 h, 0.01 mg/mL showed mild larvicidal effects, and 0.05 mg/mL showed high effects, with survival rates of 42% and 23%, respectively. The LC-50 at 72 h was noted to be 9 µg/mL.
Increasing both the catechin concentration and the exposure time showed reduced survival rates of D. magna and A. salina. Catechin hydrate was more toxic, likely due to its pure form and greater concentration of active compounds, resulting in stronger biological effects. Its higher solubility and bioavailability further enhance absorption by organisms, which intensifies its toxic impact. In contrast, crude catechin, being a crude extract, contains a mixture of various compounds, some of which may function as buffers or neutralizers of toxic effects [22,39]. The absence of these mitigating components in catechin hydrate could contribute to its more pronounced toxicity in zooplankton.
The larvicidal effect of crude catechin on A. salina was higher than that reported in Rodrigues et al. [39], who reported no toxic effects of Camellia sinensis L. Kuntze (green tea) on A. salina up to 1.0 mg/mL for 48 h, but lower than that reported in Silva et al. [22], who reported mild larvicidal effects of matcha green tea extract (C. sinensis), with an LC-50 at 24 h of 0.4 mg/mL. In addition, they observed an increase in mortality with increasing concentration (up to 1000 μg/mL) of matcha green tea extract. The larvicidal effects of crude catechin and catechin hydrate on A. salina appeared to be weaker than that on D. magna. This is probably because Daphnia sp. is more sensitive to environmental toxicity than Artemia sp., as shown by Ivorra et al. [40] and de la Vega et al. [15]. Harwanto et al. [16] also reported higher mortality in D. magna than in A. salina when exposed to the same concentrations of crude phlorotannin or pure phlorotannin.
Crude catechin also impacted L. sativa germination and radicle growth. Germination was relatively higher at 1.0 mg/mL (87.5 ± 15.0%) but decreased with higher concentrations, with significant inhibition observed at 50.0 mg/mL, resulting in 0% germination. Radicle growth was optimal at 1.0 mg/mL (18.02 ± 3.42 mm) and minimal at 10.0 mg/mL (4.71 ± 1.03 mm). Catechin hydrate led to faster radicle growth at 0.1 mg/mL (29.75 ± 1.00 mm at 96 h). The germination rate decreased with increasing concentrations, with the highest rate at 0.5 mg/mL (90.0 ± 0.00%) and the lowest at 10.0 mg/mL (33.3 ± 15.3%) at 96 h. This pattern suggests that higher catechin levels exert an inhibitory effect on early plant development. However, despite prolonged exposure, both germination rate and radical length remained elevated, indicating a potential resilience or adaptive physiological response in L. sativa, enabling partial recovery or continued growth even under suboptimal conditions. A previous study by Harwanto et al. [16] using phlorotannin and phloroglucinol showed lower survival rates for L. sativa compared to the present study, suggesting a lower toxicity of crude catechin and catechin hydrate. In this study, radical growth was slower for crude catechin but faster for catechin hydrate compared to phlorotannin and phloroglucinol.
Crude catechin inhibited the cell density of freshwater C. vulgaris at all tested concentrations after 24 h. At 48 h, the lowest concentration of catechin hydrate did not show significant changes. At 72 h, 0.5 mg/mL crude catechin and 0.05 and 0.1 mg/mL catechin hydrate increased cell density by 4.61%, 8.10% and 2.91%, respectively. This study found that C. vulgaris cell density decreased with increasing catechin concentration, indicating an inhibitory effect of catechin exposure. However, cell density continued to rise with longer exposure, suggesting a potential tolerance of C. vulgaris to catechin exposure. At the lowest catechin concentration, cell density was higher than in the control, indicating a potential stimulatory effect of catechin. Rashidinejad et al. [14] reported comparable results, showing that minimal doses of catechins can improve health, while higher doses can be toxic. Seawater C. vulgaris exposed to crude catechin and catechin hydrate showed similar patterns. Concentrations of 0.5 mg/mL crude catechin and 0.05 and 0.1 mg/mL catechin hydrate increased cell density by 2.61%, 5.92%, and 1.71%, respectively. A previous study by Harwanto et al. [16] also reported higher toxicity for phlorotannin and phloroglucinol compared to crude catechins and catechin hydrate. The results of the present study suggest that crude catechins and catechin hydrates can act as fertilizers at lower concentrations.
Catechins can induce oxidative stress by promoting the generation of reactive oxygen species (ROS), leading to cellular damage in various biological systems. This oxidative stress affects critical cellular components such as DNA, proteins, and lipids. In aquatic organisms like D. magna and A. salina, this oxidative stress may contribute to the observed toxicity, as both species serve as valuable models for assessing the overall cytotoxicity of compounds [15,16,17,18,19,21,22,23,24,27,40]. These crustaceans are commonly used to evaluate the potential toxic effects of chemicals, providing insights into the compound’s impact on multiple biological systems within an organism. The primary target organs and tissues for catechins in D. magna and A. salina include the digestive system, nervous system, reproductive organs, and epidermal tissues. The effects of catechins on these organs suggest that they may disrupt a wide range of physiological processes, leading to broader systemic toxicity. The observed increase in mortality of D. magna and A. salina with increasing catechin concentration and exposure time in this study is strongly suspected to be associated with elevated levels of ROS, which are known to induce cellular damage across various biological systems [21,22,23,24,27,40]. This finding is consistent with previous studies demonstrating that ROS levels tend to rise in response to higher concentrations and prolonged exposure to a variety of chemical agents. Similar patterns of ROS-induced toxicity have been reported with exposure to nano-titanium dioxide [15], extract of matcha green tea [22], dental restoration materials [23], and extract of Hyptis suaveolens [24].
Catechin compounds exhibited a range of phytotoxic effects on L. sativa and C. vulgaris primarily through their interaction with ROS and their influence on cellular processes. These compounds can act as both protective agents and phytotoxins, depending on their concentration. As phytotoxins, catechins induce ROS production, leading to oxidative stress and cell death in plants and algae. In L. sativa, the root system is a primary target of phytotoxins, where catechins or other phytotoxic compounds induce ROS production, leading to root cell death and inhibition of root growth [31,32,33,34,41]. In C. vulgaris, phytotoxins primarily target body cells or newly developing cells, especially those involved in cell division or growth processes [16,35,42]. Phytotoxins like those derived from catechins can induce ROS production, causing oxidative stress, damaging cell membranes, and ultimately disrupting cellular functions. They also affect chloroplasts, the main organelles responsible for photosynthesis, which can lead to cell death.
Research on the effects of catechins on aquatic organisms holds enormous potential to provide insights into how these natural compounds can impact aquatic ecosystems and environmental quality. Catechins are known for their various biological benefits, such as antioxidants, antimicrobial, and anti-inflammatory properties [1,2,3,4,5,6,7,8]. However, their effects on aquatic microorganisms, such as bacteria, algae, and protozoa, can vary significantly depending on the concentration, type of microorganism, and environmental quality. Some studies suggest that catechins have the potential to inhibit the growth of certain microorganisms, functioning as natural antimicrobial agents [22,39]. However, at certain concentrations, catechins may disrupt the microbial balance within the aquatic ecosystem, either by reducing microbial diversity or affecting specific populations crucial for biogeochemical cycles. This research demonstrates that crude catechins and catechin hydrate can influence the presence of zooplankton and algae, which are foundational to the aquatic food chain. Their impact on algae, whether through stimulating or inhibiting growth, affects oxygen production in the water and the stability of the aquatic ecosystem. A reduction in zooplankton populations could disrupt the ecological balance and decrease food availability for other organisms in the aquatic food web. The study also indicates that if catechins are applied in high concentrations or over extended periods [38,39], their effects on aquatic microorganisms may become more pronounced. If microorganisms beneficial to water quality, such as organic matter decomposers, are affected, it could interfere with processes like organic matter decomposition and nitrification, both of which are crucial for maintaining nutrient balance in aquatic environments.
It is important to note that further research is needed to better understand the bioaccumulation of catechins in the tissues of microorganisms, as well as the long-term impacts on aquatic microorganisms and the entire ecosystem. If catechins are used in large quantities, potential risks may include changes in the microbial community structure within aquatic environments, ultimately affecting the health and sustainability of the overall aquatic ecosystem. Additionally, the interactions between catechins and other pollutants or chemicals in the water need further exploration, as these compounds may have synergistic or antagonistic effects that could either exacerbate or improve environmental conditions.

4. Conclusions

This study demonstrates that the survival rate of zooplankton in both freshwater and seawater decreases with higher concentrations of and longer exposure times to catechin. In plants such as L. sativa, germination and radicle growth can still increase with extended catechin exposure, similar to the response in microalgae like C. vulgaris, which can increase its cell density even as catechin exposure time extends. Chlorella vulgaris can even grow faster than the control at low catechin concentrations, indicating a stimulatory effect of catechins. The use of A. salina, D. magna, L. sativa, and C. vulgaris in toxicity tests confirmed the larvicidal and inhibitory effects of various concentrations of crude catechins and catechin hydrate. Crude catechin and catechin hydrate were more larvicidal for D. magna than A. salina, while the inhibitory effect was greater in seawater C. vulgaris compared to freshwater C. vulgaris. The results of this study suggest that catechins, at appropriate concentrations, can function as fertilizers, enhancing the growth of terrestrial plants and microalgae.

5. Materials and Methods

5.1. Sample Preparation

Daphnia magna Straus were obtained from a local live feed breeder for fish. Adult D. magna were reared in a 2 L Erlenmeyer flask in the laboratory, with the rearing medium maintained at 25 ± 1.0 °C and aeration provided for oxygen supply. During rearing, C. vulgaris was provided daily as feed. D. magna neonates aged 24 h were harvested for use as test animals, following previous studies [16,43].
Dried brine shrimp eggs (Artemia salina L.; SWORM, Seoul, Republic of Korea) were placed in a 250 mL flask containing filtered seawater (32 ppt), with the density of dry brine shrimp eggs set at 1.0 g egg/L. The flasks were incubated at 20 °C with a constant light intensity of 40 μmol/m2/s and slight aeration. Artemia salina was harvested using a Pasteur pipette 24 h post-hatching for bioassays [16,27].
Lettuce seeds (Lactuca sativa L.; Korean cultivar, Hongbitjeokchimasangchu) were purchased from Danong Co., Ltd., Namyangju, Republic of Korea. Seeds were selected based on their intact profile, undamaged condition, and uniform size for growth and survival monitoring.
Chlorella vulgaris Beijerinck culture (FBCC-A49) was obtained from the Bioresources Culture Collection, Sangju, Republic of Korea, and divided into two portions for cultivation in freshwater and seawater (32 ppt) medium. A 250 mL Erlenmeyer flask containing 150 mL of Bold’s Basal Medium (BBM) was used as the culture medium [35,42]. Dense cultures were diluted with new sterile BBM every four days to maintain exponential growth. Cultivation took place in a shaking incubator (VS-8480; KOLAB, Korea Science Ltd., Uiwang, Republic of Korea) at 150 rpm, 23.0 ± 1.0 °C, with a light: dark cycle of 14:10 and a light intensity of 4000 lx, under aseptic conditions to avoid contamination.

5.2. Catechin Hydrate and Catechin Extracts

Catechin hydrate with 98% purity (HPLC grade) was purchased from Sigma Aldrich, St. Louis, MO, USA. Crude catechins used in this study were extracted from food-grade semi-purified green tea extract powder (Anhui Redstar Pharmaceutical Corp., Ltd., Xuan, Anhui, China) following the method reported by Vuong et al. [44]. Briefly, green tea powder was extracted into a solvent, concentrated, separated, isolated, and dried to produce crude catechins in powder form for further analysis. Catechin hydrate and crude catechin extracts at different concentrations were used to treat A. salina nauplii, D. magna neonates, L. sativa seeds, and C. vulgaris. The determination of concentration was based on the preliminary experiments. Crude catechin showed a lower toxic level compared to catechin hydrate. The effects on each organism varied, as did the duration of exposure required. Based on these factors, the concentration applied to arthropods and the observation period differed from those applied to L. sativa and C. vulgaris.

5.3. HPLC Analysis

The catechin content in the green tea extract from Anhui Redstar Pharmaceutical Corporation was analyzed using a 2424 Evaporative Light Scattering (ELC) Detector integrated with an Alliance HPLC System (Waters Corp., Milford, MA, USA) and a Platinum EPS C18 reversed-phase column (53 × 7 mm, 1.5 μm) from Analytical (Flanders, NJ, USA). Standard catechin was sourced from Sigma Aldrich, St. Louis, MO, USA. The mobile phase comprised water and acetonitrile at an 87:13 ratio, with 0.05% (v/v) trifluoroacetic acid. Samples were filtered through a 0.20 µm syringe filter (Agilent Technologies, Inc., Santa Clara, CA, USA) prior to injection, and the mobile phase was filtered with a 0.45 µm membrane filter (Agilent Technologies) before use. Catechin detection was performed at 210 nm. The sample injection volume was set at 20 µL with a flow rate of 1.0 mL/min, and results were quantified by comparing them with those of the standard solution.

5.4. A. salina and D. magna Lethality Test

The A. salina and D. magna were placed in 24-well plates modified to supply oxygen, based on a previous study by Harwanto et al. [16], as an external oxygen supply is necessary at high densities. The mortality test method for D. magna and A. salina in this study was based on previous similar studies [16,27,43], with slight modifications. The lethality test was performed to determine the toxic effects of exposure to crude catechins and catechin hydrate on D. magna and A. salina.
Twenty D. magna neonates were placed in each well containing 400 µL of distilled freshwater media, while 20 A. salina nauplii were placed in each well containing 400 µL of sterilized seawater media. Concentrations of crude catechins and catechin hydrate were 0.5, 1.0, 2.0, and 3.0 mg/mL for crude catechins and 0.01, 0.05, 0.1, and 0.5 mg/mL for catechin hydrate. Controls without crude catechin or catechin hydrate were also included. The media volume in each well was adjusted to 2 mL with freshwater or seawater [27]. Plates were maintained at 24 ± 1 °C. Survival rate observations were conducted at 24, 48, and 72 h. All experiments were repeated three times.
The survival rate was based on criteria specified by Kim and Choi [27] and Harwanto et al. [16]. Daphnia magna neonates and A. salina nauplii were considered dead if they showed no movement during observation. Concentration was classified as non-larvicide if the survival rate was >50%, mild larvicidal if >25% but <50%, highly larvicidal if <25%, and extremely larvicidal if no organisms survived.

5.5. L. sativa Germination Assays

The germination analysis followed the methods of Choi and Choi [41] and Harwanto et al. [16]. Filter papers with a diameter of 55 mm (Toyo Roshi Kaisha, Ltd., Tokyo, Japan) were placed in 6 cm-diameter sterile Petri dishes and treated with crude catechin or catechin hydrate at concentrations of 1.0, 10.0, and 50.0 mg/mL. The filter papers were then air-dried to ensure that the toxic properties were derived solely from the catechins absorbed on the filter paper.
Ten intact, uniform L. sativa seeds with no broken parts were evenly placed on each filter paper and saturated with 1% Tween 20 (L33109; Duksan Pure Chemicals, Ansan, Republic of Korea). Controls were treated with only 1% Tween 20 [45,46]. Since Tween 20 is non-toxic, it was utilized as a surfactant in this study [16]. The plates were placed in an incubator at 25 ± 1 °C in the dark, and germination rates and radicle length were recorded every 24 h for four days. Seeds were considered germinated when the radicle length was 1 mm long [16], measured by a digital caliper. The survival rate of the seeds was determined based on the percentage of germinated seeds.

5.6. Yield Inhibition Test of C. vulgaris

The yield inhibition test was based on previous studies by Qian et al. [35] and Xiong et al. [47], with slight modifications. Chlorella vulgaris was placed in 15 mL vials containing BBM medium with various concentrations of crude catechin (0.5, 1.0, and 2.0 mg/mL) or catechin hydrate (0.05, 0.1, 0.5, and 1.0 mg/mL). Control vials contained BBM solution without crude catechin or catechin hydrate. Each experiment was repeated three times. Vials were incubated for 72 h under aseptic conditions to avoid contamination. The optical density (OD) was evaluated at 24, 48, and 72 h at 680 nm (OD680) using a SPECTROstar® Nano spectrophotometer (BMG LABTECH, Offenburg, Germany).
Cell density (cells/mL) was measured using a hemocytometer (HSU-0650010; Paul Marienfeld GmbH & Co. KG., Lauda-Königshofen, Germany), under a microscope. Chlorella vulgaris with a high cell density was diluted to examine the association between optical density and cell density. The linear relationship between OD680 and cell density was determined by Equation (1).
Cell density = [(OD680 + 0.0282)/0.0004] × 10,000 (R2 = 0.9936)
The relationship between OD680 and dry cell weight (DCW) was also determined following Equation (2) [16,35].
DCW = 0.1244 + 680 + 0.017 (R2 = 0.9908)
The yield inhibition rate (Iy; %) was calculated following Equation (3), as reported by Xiong et al. [47].
% Iy = ((Nc − Nt)/Nc) × 100
where Nc is the DCW in the control group and Nt is the DCW in the treatment group.

5.7. Statistical Analysis

Data are presented as mean ± standard deviation (S.D.) of three independent experiments (n = 3). Experimental results were checked for normality and homogeneity before performing the Student’s t-test analysis. The t-test was used to compare each treatment group with the control group, with statistical significance set at p < 0.05. The LC-50, which is the effective toxicant concentration capable of killing 50% of the organisms, was determined using probit analysis. This involved deriving the LC-50 value through regression of the exposed solution concentration against the percentage of organism mortality [48].

Author Contributions

Conceptualization, K.M.I.B. and J.-S.C.; methodology, K.M.I.B., H.-R.A., M.D.N.M., D.H. and J.-S.C.; software, K.M.I.B. and D.H.; validation, K.M.I.B., G.T., M.D.N.M., J.-H.S., D.H. and J.-S.C.; formal analysis, K.M.I.B., H.-R.A., B.F.S.P.N. and D.H.; investigation, B.F.S.P.N., G.T., M.D.N.M. and J.-H.S.; resources, D.H. and J.-S.C.; data curation, K.M.I.B., H.-R.A., B.F.S.P.N., G.T., M.D.N.M. and D.H.; writing—original draft preparation, K.M.I.B., H.-R.A. and D.H.; writing—review and editing, K.M.I.B., M.D.N.M., J.-H.S., D.H. and J.-S.C.; visualization, K.M.I.B., M.D.N.M., J.-H.S., D.H. and J.-S.C.; supervision, J.-S.C.; project administration, J.-S.C.; funding acquisition, K.M.I.B. and J.-S.C. All authors have read and agreed to the published version of the manuscript.

Funding

This research received no external funding.

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

The original contributions presented in this study are included in this article. Further inquiries can be directed to the corresponding authors.

Conflicts of Interest

The authors declare no conflicts of interest.

Abbreviations

The following abbreviations are used in this manuscript:
LC-50Lethal concentration 50, the effective toxicant concentration capable of killing 50% of the organisms
DCWDry cell weight
ODOptical density
BBMBold’s Basal Medium
HPLCHigh-performance liquid chromatography
ECEpicatechin
EGCEpigallocatechin
ECGEpicatechin gallate
EGCGEpigallocatechin gallate

References

  1. Fathima, A.; Rao, J.R. Selective toxicity of catechin—A natural flavonoid towards bacteria. Appl. Microbiol. Biotechnol. 2016, 100, 6395–6402. [Google Scholar] [CrossRef] [PubMed]
  2. Al-Hazzani, A.A.; Alshatwi, A.A. Catechin hydrate inhibits proliferation and mediates apoptosis of SiHa human cervical cancer cells. Food Chem. Toxicol. 2011, 49, 3281–3286. [Google Scholar] [CrossRef] [PubMed]
  3. Kaur, R.; Rajput, R.; Nag, P.; Kumar, S.; Singh, M. Synthesis, characterization and evaluation of antioxidant properties of catechin hydrate nanoparticles. J. Drug Deliv. Sci. Technol. 2017, 39, 398–407. [Google Scholar] [CrossRef]
  4. Muzolf-Panek, M.; Gliszczyńska-Świgło, A.; Szymusiak, H.; Tyrakowska, B. The influence of stereochemistry on the antioxidant properties of catechin epimers. Eur. Food Res. Technol. 2012, 235, 1001–1009. [Google Scholar] [CrossRef]
  5. Kim, J.M.; Heo, H.J. The roles of catechins in regulation of systemic inflammation. Food Sci. Biotechnol. 2022, 31, 957–970. [Google Scholar] [CrossRef]
  6. Scalia, S.; Marchetti, N.; Bianchi, A. Comparative evaluation of different co-antioxidants on the photochemical- and functional-stability of epigallocatechin-3-gallate in topical creams exposed to simulated sunlight. Molecules 2013, 18, 574–587. [Google Scholar] [CrossRef]
  7. Sentkowska, A. Content of selenoaminoacids and catechins in Chinese green teas. Eur. Food Res. Technol. 2021, 247, 613–622. [Google Scholar] [CrossRef]
  8. Senanayake, S.P.J.N. Green tea extract: Chemistry, antioxidant properties and food applications—A review. J. Funct. Foods 2013, 5, 1529–1541. [Google Scholar] [CrossRef]
  9. Chan, P.T.; Fong, P.; Cheung, Y.L.; Huang, Y.; Kwok, W.; Ho, K.; Chen, Z.-Y. Biochemical and molecular action of nutrients jasmine green tea epicatechins are hypolipidemic in hamsters (Mesocricetus auratus) fed a high fat diet. J. Nutr. 1999, 129, 1094–1101. [Google Scholar] [CrossRef]
  10. Nagao, T.; Meguro, S.; Soga, S.; Otsuka, A.; Tomonobu, K.; Fumoto, S.; Chikama, A.; Mori, K.; Yuzawa, M.; Watanabe, H.; et al. Tea catechins suppress accumulation of body fat in humans. J. Oleo Sci. 2001, 5, 717–728. [Google Scholar] [CrossRef]
  11. Nagao, T.; Komine, Y.; Soga, S.; Meguro, S.; Hase, T.; Tanaka, Y.; Tokimitsu, I. Ingestion of a tea rich in catechins leads to a reduction in body fat and malondialdehyde-modified LDL in men. Am. J. Clin. Nutr. 2005, 81, 122–129. [Google Scholar] [CrossRef] [PubMed]
  12. Nagao, T.; Hase, T.; Tokimitsu, I. A green tea extract high in catechins reduces body fat and cardiovascular risks in humans. Obesity 2007, 15, 1473–1483. [Google Scholar] [CrossRef] [PubMed]
  13. Fang, Z.; Zhao, J.; Li, Y.; Wang, Y.; Qiu, T.; Wu, Y.; Dong, W.; Mailhot, G. Improving Fenton-like system with catechin, an environmental-friendly polyphenol: Effects and mechanism. Chem. Eng. J. 2021, 426, 127946. [Google Scholar] [CrossRef]
  14. Rashidinejad, A.; Boostani, S.; Babazadeh, A.; Rehman, A.; Rezaei, A.; Akbari-Alavijeh, S.; Shaddel, R.; Jafari, S.M. Opportunities and challenges for the nanodelivery of green tea catechins in functional foods. Food Res. Int. 2001, 142, 110186. [Google Scholar] [CrossRef]
  15. De la Vega, A.C.S.; Cruz-Alcalde, A.; Mazón, C.S.; Martí, C.B.; Diaz-Cruz, M.S. Nano-TiO2 phototoxicity in fresh and seawater: Daphnia magna and Artemia sp. as proxies. Water 2021, 13, 55. [Google Scholar] [CrossRef]
  16. Harwanto, D.; Negara, B.F.S.P.; Tirtawijaya, G.; Meinita, M.D.N.; Choi, J.S. Evaluation of toxicity of crude phlorotannins and phloroglucinol using different model organisms. Toxins 2022, 14, 312. [Google Scholar] [CrossRef]
  17. Fuertes, I.; Piña, B.; Barata, C. Changes in lipid profiles in Daphnia magna individuals exposed to low environmental levels of neuroactive pharmaceuticals. Sci. Total Environ. 2020, 733, 139029. [Google Scholar] [CrossRef]
  18. Szabelak, A.; Bownik, A. Behavioral and physiological responses of Daphnia magna to salicylic acid. Chemosphere 2021, 270, 128660. [Google Scholar] [CrossRef]
  19. Adamczuk, M. Environmentally realistic concentrations of ibuprofen influence life histories but not population dynamics of Daphnia magna. Sci. Total Environ. 2022, 848, 157783. [Google Scholar] [CrossRef]
  20. Cristóvão, M.B.; Bernardo, J.; Bento-Silva, A.; Ressureição, M.; Bronze, M.R.; Crespo, J.G.; Pereira, V.J. Treatment of anticancer drugs in a real wastewater effluent using nanofiltration: A pilot scale study. Sep. Purif. Technol. 2022, 288, 1206565. [Google Scholar] [CrossRef]
  21. Tkaczyk, A.; Bownik, A.; Dudka, J.; Kowal, K.; Ślaska, B. Daphnia magna model in the toxicity assessment of pharmaceuticals: A review. Sci. Total Environ. 2021, 763, 143038. [Google Scholar] [CrossRef] [PubMed]
  22. Silva, T.M.; Fracasso, D.S.; Visentin, A.P.V.; Cassini, C.; Scariot, F.J.; Danetti, S.; Echeverrigaray, S.; Moura, S.; Touguinha, L.B.; Branco, C.S.; et al. Dual effect of the herbal matcha green tea (Camellia sinensis L. Kuntze) supplement in EA.hy926 endothelial cells and Artemia salina. J. Ethnopharmacol. 2022, 298, 115564. [Google Scholar] [CrossRef] [PubMed]
  23. Milhem, M.M.; Al-Hiyasat, A.S.; Darmani, H. Toxicity testing of restorative dental materials using brine shrimp larvae (Artemia salina). J. Appl. Oral Sci. 2008, 16, 297–301. [Google Scholar] [CrossRef] [PubMed]
  24. Bezerra, J.W.A.; Costa, A.R.; da Silva, M.A.P.; Rocha, M.I.; Boligon, A.A.; da Rocha, J.B.T.; Barros, L.M.; Kamdem, J.P. Chemical composition and toxicological evaluation of Hyptis suaveolens (L.) Poiteau (LAMIACEAE) in Drosophila melanogaster and Artemia salina. S. Afr. J. Bot. 2017, 113, 437–442. [Google Scholar] [CrossRef]
  25. Minello, L.; Mattiello, G.S.; Cassini, C.; Touguinha, L.A.; Paim, L.F.; Moura, S.; Salvador, M.; Branco, C.S. Chemical characterization, in vivo safety profile and effects of the extract from unconventional food plant Acca sellowiana (O. Berg) Burret on endothelial cells under glucose overload. J. Pharm. Biomed. Anal. 2024, 248, 116304. [Google Scholar] [CrossRef]
  26. Oliveira, M.A.; dos Santos, C.A.L.; da Silva Teles, B.R.; Oliveira, C.V.B.; da Silva, V.B.; Pereira, A.L.G.; Rodrigues, V.L.C.; Fonseca, V.J.A.; dos Santos Santana, M.; Lima, C.M.G.; et al. Chemical composition, antifungal activity and toxicological evaluation of Lippia sidoides Cham. J. Agric. Food Res. 2024, 18, 101333. [Google Scholar] [CrossRef]
  27. Kim, Y.D.; Choi, J.-S. Larvicidal effects of Korean seaweed extracts on brine shrimp Artemia salina. J. Anim. Plant Sci. 2017, 27, 1039–1046. [Google Scholar]
  28. Balamurugan, V.; Ragavendran, C.; Arulbalachandran, D.; Alrefaei, A.F.; Rajendran, R. Green synthesis of silver nanoparticles using Pandanus tectorius aerial root extract: Characterization, antibacterial, cytotoxic, and photocatalytic properties, and ecotoxicological assessment. Inorg. Chem. Commun. 2024, 168, 112882. [Google Scholar] [CrossRef]
  29. Tigre, R.C.; Silva, N.H.; Santos, M.G.; Honda, N.K.; Falcão, E.P.S.; Pereira, E.C. Allelopathic and bioherbicidal potential of Cladonia verticillaris on the germination and growth of Lactuca sativa. Ecotoxicol. Environ. Saf. 2012, 84, 125–132. [Google Scholar] [CrossRef]
  30. Bertan, A.S.; Teles, A.G.D.X.; de Oliveira, B.C.T.; Ferreira, L.M.R.; Thiel, N.A.; Fieira, C.; Chiarelotto, M.; Manosso, F.C.; Pokrywiecki, J.C.; Tonial, I.B.; et al. Influence of the seasons on the water quality of the Marmeleiro River (Paraná, Brazil) using different bioindicators. Discov. Appl. Sci. 2020, 2, 1825. [Google Scholar] [CrossRef]
  31. Guzmán-Morales, A.R.; Cruz-La Paz, O.; Valdés-Carmenate, R.; Valdés-Hernández, P.A. Evaluation of heavy metal contamination and accumulation in lettuce (Lactuca sativa L.) plants. Cult. Trop. 2021, 42, e03. [Google Scholar]
  32. Izquierdo-Díaz, M.; Hansen, V.; Barrio-Parra, F.; De Miguel, E.; You, Y.; Magid, J. Assessment of lettuces grown in urban areas for human consumption and as bioindicators of atmospheric pollution. Ecotoxicol. Environ. Saf. 2023, 256, 114883. [Google Scholar] [CrossRef] [PubMed]
  33. De Moura, D.R.; Araujo, E.C.G.; Borges, C.H.A.; Araujo, L.H.B.; de Lima, T.V.; Silva, T.C.; da Nobrega, C.C. Alellopathic effects of aqueous extracts from Bambusa vulgaris Schrad. Ex J. C. Wendl. on seed germination and vigor from Lactuca sativa. Afr. J. Agric. Res. 2018, 13, 1954–1958. [Google Scholar]
  34. Souza, M.M.V.; Souza, A.D.V.; Rodrigues, A.A.; Batista, P.F.; Castro, S.T.; Silva, I.M.H.L.; Jakelaitis, A.; Costa, A.C.; Sales, J.F. The allelopathic effects of aqueous Talinum triangulare (Jacq.) wild extracts on the development of Lactuca sativa L. seedlings. Braz. J. Biol. 2024, 84, e279983. [Google Scholar] [CrossRef]
  35. Qian, L.; Qi, S.; Cao, F.; Zhang, J.; Zhao, F.; Li, C.; Wang, C. Toxic effects of boscalid on the growth, photosynthesis, antioxidant system and metabolism of Chlorella vulgaris. Environ. Pollut. 2018, 242, 171–181. [Google Scholar] [CrossRef]
  36. Hu, J.; Webster, D.; Cao, J.; Shao, A. The safety of green tea and green tea extract consumption in adults—Results of a systematic review. Regul. Toxicol. Pharmacol. 2018, 95, 412–433. [Google Scholar] [CrossRef]
  37. Chobot, V.; Huber, C.; Trettenhahn, G.; Hadacek, F. (±)-Catechin: Chemical weapon, antioxidant, or stress regulator? J. Chem. Ecol. 2009, 35, 980–996. [Google Scholar] [CrossRef]
  38. Ahmad, N.; Ahmad, R.; Alrasheed, R.A.; Almatar, H.M.A.; Al-Ramadan, A.S.; Amir, M.; Sarafroz, M. Quantification and evaluations of catechin hydrate polymeric nanoparticles used in brain targeting for the treatment of epilepsy. Pharmaceutics 2020, 12, 203. [Google Scholar] [CrossRef]
  39. Rodrigues, M.J.; Neves, V.; Martins, A.; Rauter, A.P.; Neng, N.R.; Nogueira, J.M.F.; Varela, J.; Barreira, L.; Custódio, L. In vitro antioxidant and anti-inflammatory properties of Limonium algarvense flowers’ infusions and decoctions: A comparison with green tea (Camellia sinensis). Food Chem. 2016, 200, 322–329. [Google Scholar] [CrossRef]
  40. Ivorra, L.; Cardoso, P.G.; Chan, S.K.; Tagulao, K.; Cruzeiro, C. Environmental characterization of 4,4′-dichlorobenzophenone in surface waters from Macao and Hong Kong coastal areas (Pearl River Delta) and its toxicity on two biological models: Artemia salina and Daphnia magna. Ecotoxicol. Environ. Saf. 2019, 171, 1–11. [Google Scholar] [CrossRef]
  41. Choi, J.-S.; Choi, I.S. Inhibitory effect of marine green algal extracts on the germination of Lactuca sativa seeds. J. Environ. Biol. 2016, 37, 207–213. [Google Scholar] [PubMed]
  42. Xiong, J.Q.; Kurade, M.B.; Jeon, B.H. Biodegradation of levofloxacin by an acclimated freshwater microalga, Chlorella vulgaris. Chem. Eng. J. 2017, 313, 1251–1257. [Google Scholar] [CrossRef]
  43. Na, J.; Kim, Y.; Song, J.; Shim, T.; Cho, K.; Jung, J. Evaluation of the combined effect of elevated temperature and cadmium toxicity on Daphnia magna using a simplified DEBtox model. Environ. Pollut. 2021, 291, 118250. [Google Scholar] [CrossRef] [PubMed]
  44. Vuong, Q.V.; Golding, J.B.; Nguyen, M.; Roach, P.D. Extraction and isolation of catechins from tea. J. Sep. Sci. 2010, 33, 3415–3428. [Google Scholar] [CrossRef]
  45. Vinayak, R.C.; Sudha, S.A.; Chatterji, A. Bio-screening of a few green seaweeds from India for their cytotoxic and antioxidant potential. J. Sci. Food Agric. 2011, 91, 2471–2476. [Google Scholar] [CrossRef]
  46. Kang, S.M.; Khan, A.L.; Hussain, J.; Ali, L.; Kamran, M.; Waqas, M.; Lee, I.J. Rhizonin A from Burkholderia sp. KCTC11096 and its growth promoting role in lettuce seed germination. Molecules 2012, 17, 7980–7988. [Google Scholar] [CrossRef]
  47. Xiong, J.Q.; Kurade, M.B.; Abou-Shanab, R.A.I.; Ji, M.K.; Choi, J.; Kim, J.O.; Jeon, B.H. Biodegradation of carbamazepine using freshwater microalgae Chlamydomonas mexicana and Scenedesmus obliquus and the determination of its metabolic fate. Bioresour. Technol. 2016, 205, 183–190. [Google Scholar] [CrossRef]
  48. Beula, M.; Ravikumar, S.; Ali, M.S. Mosquito larvicidal efficacy of seaweed extracts against dengue vector of Aedes aegypti. Asian Pac. J. Trop. Biomed. 2011, 1, S143–S146. [Google Scholar] [CrossRef]
Toxins 17 00244 g001
Figure 1. HPLC chromatogram of standard catechin at 50 µg/mL (a), crude catechin at 200 µg/mL (b), and combination of crude catechin (200 µg/mL) + standard catechin (50 µg/mL) (c).
Figure 1. HPLC chromatogram of standard catechin at 50 µg/mL (a), crude catechin at 200 µg/mL (b), and combination of crude catechin (200 µg/mL) + standard catechin (50 µg/mL) (c).
Toxins 17 00244 g001
Table 1. Larvicidal effects of crude catechin and catechin hydrate on the survival rate of D. magna neonates.
Table 1. Larvicidal effects of crude catechin and catechin hydrate on the survival rate of D. magna neonates.
Incubation Time (h)Survival Rate (%) of Daphnia magna Straus NeonatesLC-50
(µg/mL)
ControlCrude Catechin (mg/mL)
0.51.02.03.0
24100.00 ± 0.0073.00 ± 2.90 *64.00 ± 6.30 *45.00 ± 4.10 *10.00 ± 4.10 *1174
4894.00 ± 7.5043.00 ± 8.70 *10.00 ± 8.20 *0.00 ± 0.00 *0.00 ± 0.00 *
7285.00 ± 7.1028.00 ± 5.00 *4.00 ± 2.50 *0.00 ± 0.00 *0.00 ± 0.00 *
Incubation Time (h)ControlCatechin Hydrate (mg/mL)LC-50
(µg/mL)
0.010.050.10.5
24100.00 ± 0.0088.00 ± 2.90 *68.00 ± 7.60 *38.00 ± 2.90 *0.00 ± 0.00 *54
4892.00 ± 2.9070.00 ± 5.00 *55.00 ± 5.00 *28.00 ± 2.90 *0.00 ± 0.00 * 39
7277.00 ± 2.9040.00 ± 5.00 *20.00 ± 5.00 *7.00 ± 2.90 *0.00 ± 0.00 *7
Values are mean ± S.D. of at least three replications, where each replication contains 20 entities. LC-50: lethal concentration 50, the effective toxicant concentration capable of killing 50% of the organisms (confidence level of 95%). * Indicates significant differences based on Student’s t-test at p < 0.05 compared to control.
Table 2. Larvicidal effects of crude catechin and catechin hydrate on the survival rate of A. salina nauplii.
Table 2. Larvicidal effects of crude catechin and catechin hydrate on the survival rate of A. salina nauplii.
Incubation Time (h)Survival Rate (%) of Artemia salina L. NaupliiLC-50
(µg/mL)
ControlCrude Catechin (mg/mL)
0.51.02.03.0
24100.00 ± 0.00100.00 ± 0.0099.00 ± 2.5054.00 ± 8.50 *16.00 ± 4.80 *1895
4898.00 ± 2.9085.00 ± 4.10 *20.00 ± 4.10 *0.00 ± 0.00 *0.00 ± 0.00 *
7293.00 ± 2.9033.00 ± 8.70 *13.00 ± 6.50 *0.00 ± 0.00 *0.00 ± 0.00 *
Incubation Time (h)ControlCatechin hydrate (mg/mL)LC-50
(µg/mL)
0.010.050.10.5
24100.00 ± 0.0090.00 ± 0.00 *63.00 ± 2.90 *45.00 ± 5.00 *40.00 ± 5.00 *153
4897.00 ± 2.9080.00 ± 2.90 *52.00 ± 7.60 *37.00 ± 5.80 *28.00 ± 2.90 *70
7283.00 ± 2.9042.00 ± 2.90 *23.00 ± 2.90 *10.00 ± 10.00 *0.00 ± 0.00 *9
Values are mean ± S.D. of at least three replications, where each replication contains 20 entities. LC-50: lethal concentration 50, the effective toxicant concentration capable of killing 50% of the organisms (confidence level of 95%). * Indicates significant differences based on Student’s t-test at p < 0.05 compared to control.
Table 3. Inhibitory effects of crude catechin and catechin hydrate on the germination rate (%) and radical length (mm) of L. sativa seeds.
Table 3. Inhibitory effects of crude catechin and catechin hydrate on the germination rate (%) and radical length (mm) of L. sativa seeds.
Incubation Time (h)Germination Rate (%) of Lactuca sativa L. Seeds (Radicle Length of L. sativa Seeds in mm)
ControlCrude Catechin (mg/mL)
1.010.050.0
2482.50 ± 12.6087.50 ± 15.0035.00 ± 10.00 *0.00 ± 0.00 *
(3.84 ± 0.46)(3.11 ± 0.50)(1.95 ± 0.39 *)(0.00 ± 0.00 *)
4890.00 ± 8.1695.00 ± 10.0055.00 ± 5.80 *0.00 ± 0.00 *
(11.34 ± 2.66)(7.53 ± 1.25)(3.08 ± 0.41 *)(0.00 ± 0.00 *)
7295.00 ± 5.8097.50 ± 5.0075.00 ± 5.80 *0.00 ± 0.00 *
(17.72 ± 3.57)(12.67 ± 2.27)(3.64 ± 0.56 *)(0.00 ± 0.00 *)
9695.00 ± 5.8097.50 ± 5.0075.00 ± 5.80 *0.00 ± 0.00 *
(23.61 ± 4.64)(18.02 ± 3.42)(4.71 ± 1.03 *)(0.00 ± 0.00 *)
Incubation Time (h)ControlCatechin Hydrate (mg/mL)
0.10.51.05.010.0
2483.30 ± 5.8073.30 ± 5.8073.30 ± 5.8070.00 ± 10.0060.00 ± 17.3016.70 ± 11.60 *
(4.10 ± 0.43)(4.47 ± 0.07)(4.79 ± 0.55)(4.42 ± 0.37)(2.95 ± 0.41 *)(1.89 ± 0.08 *)
4890.00 ± 10.0083.30 ± 5.8080.00 ± 10.0070.00 ± 10.0073.30 ± 20.8033.30 ± 15.30 *
(15.74 ± 1.60)(13.26 ± 0.62 *)(13.40 ± 1.48)(12.95 ± 2.31)(4.33 ± 0.56 *)(2.40 ± 0.39 *)
7293.30 ± 5.8083.30 ± 5.8086.70 ± 5.8083.30 ± 15.3090.00 ± 10.0033.30 ± 15.30 *
(24.54 ± 2.48)(21.93 ± 1.84)(20.80 ± 3.52)(15.56 ± 1.31 *)(5.85 ± 0.92 *)(2.63 ± 0.60 *)
9693.30 ± 5.8083.30 ± 5.8090.00 ± 0.0083.30 ± 15.3090.00 ± 10.0033.30 ± 15.30 *
(33.03 ± 1.56)(29.75 ± 1.00 *)(26.53 ± 1.23 *)(20.23 ± 3.14 *)(6.08 ± 0.27 *)(2.72 ± 0.62 *)
Values are mean ± S.D. of at least three replications, where each replication contains 10 entities. * Indicates significant differences based on Student’s t-test at p < 0.05 compared to control.
Table 4. Toxicity of crude catechin and catechin hydrate on the cell density of freshwater C. vulgaris.
Table 4. Toxicity of crude catechin and catechin hydrate on the cell density of freshwater C. vulgaris.
Incubation Time (h)Cell Density of Freshwater Chlorella vulgaris Beijerinck (106 cells/mL)
ControlCrude Catechin (mg/mL)
0.51.02.0
01.08 ± 0.021.05 ± 0.061.05 ± 0.061.06 ± 0.01
241.16 ± 0.051.50 ± 0.04 *1.42 ± 0.06 *1.17 ± 0.06
482.02 ± 0.082.33 ± 0.07 *2.27 ± 0.07 *1.50 ± 0.04 *
723.71 ± 0.243.84 ± 0.103.31 ± 0.22 *1.58 ± 0.05 *
964.26 ± 0.144.60 ± 0.17 *3.40 ± 0.08 *1.90 ± 0.06 *
Yield inhibition−(4.61) ± 0.83%12.29 ± 1.60%33.84 ± 0.60%
Incubation Time (h)ControlCatechin Hydrate (mg/mL)
0.050.10.51.0
01.11 ± 0.051.13 ± 0.041.10 ± 0.061.12 ± 0.061.15 ± 0.05
241.28 ± 0.041.59 ± 0.10 *1.56 ± 0.29 *1.56 ± 0.07 *1.33 ± 0.02 *
482.02 ± 0.092.22 ± 0.272.09 ± 0.282.39 ± 0.04 *1.64 ± 0.06 *
723.56 ± 0.223.99 ± 0.33 *3.61 ± 0.352.82 ± 0.07 *1.64 ± 0.05 *
964.04 ± 0.184.74 ± 0.16 *4.28 ± 0.18 *3.10 ± 0.06 *1.68 ± 0.05 *
Yield inhibition−(8.10) ± 1.22%−(2.91) ± 1.03%13.85 ± 1.53%34.91 ± 1.26%
Values are mean ± S.D. of at least three replications. * Indicates significant differences based on Student’s t-test at p < 0.05 compared to control.
Table 5. Toxicity of crude catechin and catechin hydrate on the cell density of seawater C. vulgaris.
Table 5. Toxicity of crude catechin and catechin hydrate on the cell density of seawater C. vulgaris.
Incubation Time
(h)		Cell Density of Seawater Chlorella vulgaris Beijerinck (106 cells/mL)
ControlCrude Catechin (mg/mL)
0.51.02.0
01.06 ± 0.041.02 ± 0.041.04 ± 0.041.06 ± 0.05
241.18 ± 0.041.22 ± 0.041.36 ± 0.181.23 ± 0.08
481.77 ± 0.081.90 ± 0.08 *1.97 ± 0.13 *1.37 ± 0.25 *
723.06 ± 0.213.52 ± 0.26 *2.49 ± 0.25 *1.55 ± 0.20 *
963.96 ± 0.194.25 ± 0.19 *2.63 ± 0.19 *1.61 ± 0.16 *
Yield inhibition−(2.61) ± 1.21%19.90 ± 2.29%35.26 ± 1.06%
Incubation Time
(h)		ControlCatechin Hydrate (mg/mL)
0.050.10.51.0
01.12 ± 0.071.15 ± 0.061.09 ± 0.041.15 ± 0.091.12 ± 0.13
241.25 ± 0.041.36 ± 0.11 *1.56 ± 0.22 *1.35 ± 0.09 *1.32 ± 0.08 *
482.29 ± 0.082.21 ± 0.102.20 ± 0.171.91 ± 0.12 *1.46 ± 0.09 *
723.35 ± 0.163.57 ± 0.14 *3.41 ± 0.252.28 ± 0.13 *1.72 ± 0.08
963.87 ± 0.294.15 ± 0.16 *3.98 ± 0.182.77 ± 0.09 *1.50 ± 0.09 *
Yield inhibition−(5.92) ± 2.18%−(1.71) ± 2.21%16.37 ± 1.53%35.97 ± 1.70%
Values are mean ± S.D. of at least three replications. * Indicates significant differences based on Student’s t-test at p < 0.05 compared to control.
Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content.

Share and Cite

MDPI and ACS Style

Bashir, K.M.I.; An, H.-R.; Negara, B.F.S.P.; Tirtawijaya, G.; Meinita, M.D.N.; Sohn, J.-H.; Harwanto, D.; Choi, J.-S. Toxicity Assessment of Catechins on Representative Aquatic Organisms and Terrestrial Plant. Toxins 2025, 17, 244. https://doi.org/10.3390/toxins17050244

AMA Style

Bashir KMI, An H-R, Negara BFSP, Tirtawijaya G, Meinita MDN, Sohn J-H, Harwanto D, Choi J-S. Toxicity Assessment of Catechins on Representative Aquatic Organisms and Terrestrial Plant. Toxins. 2025; 17(5):244. https://doi.org/10.3390/toxins17050244

Chicago/Turabian Style

Bashir, Khawaja Muhammad Imran, Hye-Ryeon An, Bertoka Fajar Surya Perwira Negara, Gabriel Tirtawijaya, Maria Dyah Nur Meinita, Jae-Hak Sohn, Dicky Harwanto, and Jae-Suk Choi. 2025. "Toxicity Assessment of Catechins on Representative Aquatic Organisms and Terrestrial Plant" Toxins 17, no. 5: 244. https://doi.org/10.3390/toxins17050244

APA Style

Bashir, K. M. I., An, H.-R., Negara, B. F. S. P., Tirtawijaya, G., Meinita, M. D. N., Sohn, J.-H., Harwanto, D., & Choi, J.-S. (2025). Toxicity Assessment of Catechins on Representative Aquatic Organisms and Terrestrial Plant. Toxins, 17(5), 244. https://doi.org/10.3390/toxins17050244

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

Article Metrics

Back to TopTop