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Evaluation of the neurotoxic effects of chronic embryonic exposure with inorganic mercury on motor and anxiety-like responses in zebrafish (Danio rerio) larvae

Abu Bakar N, Mohd Sata NS, Ramlan NF, Wan Ibrahim WN, Zulkifli SZ, Che Abdullah CA, et al.

Neurotoxicol Teratol, 2017 Jan-Feb;59:53-61.
PMID: 27919701 DOI: 10.1016/j.ntt.2016.11.008

Chronic exposure to mercury (Hg) can lead to cumulative impairments in motor and cognitive functions including alteration in anxiety responses. Although several risk factors have been identified in recent year, little is known about the environmental factors that either due exposure toward low level of inorganic mercury that may led to the developmental disorders. The present study investigated the effects of embryonic exposure of mercury chloride on motor function and anxiety-like behavior. The embryo exposed to 6 different concentrations of HgCl2 (7.5, 15, 30, 100, 125, 250nM) at 5hpf until hatching (72hpf) in a semi-static condition. The mortality rate increased in a dose dependent manner where the chronic embryonic exposure to 100nM decreased the number of tail coiling, heartbeat, and swimming activity. Aversive stimulus was used to examine the effects of 100nM interferes with the development of anxiety-related behavior. No elevation in both thigmotaxis and avoidance response of 6dpf larvae exposed with 100nM were found. Biochemical analysis showed HgCl2 exposure affects proteins, lipids, carbohydrates and nucleic acids of the zebrafish larvae. These results showed that implication of HgCl2 on locomotor and biochemical defects affects motor performance and anxiety-like responses. Yet, the potential underlying mechanisms these responses need to be further investigated which is crucial to prevent potential hazards on the developing organism due to neurotoxicant exposure.
Fulltext Species composition and invasion risks of alien ornamental freshwater fishes from pet stores in Klang Valley, Malaysia

Saba AO, Ismail A, Zulkifli SZ, Halim MRA, Wahid NAA, Amal MNA

Sci Rep, 2020 10 14;10(1):17205.
PMID: 33057156 DOI: 10.1038/s41598-020-74168-9

The ornamental fish trade has been considered as one of the most important routes of invasive alien fish introduction into native freshwater ecosystems. Therefore, the species composition and invasion risks of fish species from 60 freshwater fish pet stores in Klang Valley, Malaysia were studied. A checklist of taxa belonging to 18 orders, 53 families, and 251 species of alien fishes was documented. Fish Invasiveness Screening Test (FIST) showed that seven (30.43%), eight (34.78%) and eight (34.78%) species were considered to be high, medium and low invasion risks, respectively. After the calibration of the Fish Invasiveness Screening Kit (FISK) v2 using the Receiver Operating Characteristics, a threshold value of 17 for distinguishing between invasive and non-invasive fishes was identified. As a result, nine species (39.13%) were of high invasion risk. In this study, we found that non-native fishes dominated (85.66%) the freshwater ornamental trade in Klang Valley, while FISK is a more robust tool in assessing the risk of invasion, and for the most part, its outcome was commensurate with FIST. This study, for the first time, revealed the number of high-risk ornamental fish species that give an awareness of possible future invasion if unmonitored in Klang Valley, Malaysia.
Baseline distribution and sources of selected agricultural runoff in the bottom water of an active cockle farming area, Bagan Pasir, Perak, Malaysia

Joni AAM, Mohamat-Yusuff F, Noor NAM, Mohamed KN, Ash'aari ZH, Kusin FM, et al.

Mar Pollut Bull, 2021 Jun;167:112276.
PMID: 33901978 DOI: 10.1016/j.marpolbul.2021.112276

This paper aims to study the spatial and temporal patterns of selected agricultural runoff, specifically in terms of glyphosate, nitrate, and ammonia in bottom water, as well as their possible sources, within an active cockle farming area in Bagan Pasir, Perak, Malaysia. Samples were taken along the cockle farming area from March to November 2019. Glyphosate was analyzed using HPLC with both extraction and derivatization methods using 9-fluorenyl-methyl chloroformate (FMOC-Cl), while nitrate and ammonia levels were determined using the standard Hach method. Generally, glyphosate, nitrate, and ammonia were present within the study site with the average concentration of 37.44 ± 12.27 μg/l, 1.65 ± 0.52 mg/l, and 0.37 ± 0.19 mg/l, respectively. The results suggest that glyphosate and nitrate might be derived from an inland source, while a uniform and low level of ammonia suggested might originate from lithogenic origins. Continuous monitoring remains encouraged.
Toxicity effect of Diuron on gill tissue structure and the tissue residue of blood cockles (Tegillarca granosa)

Mohamat-Yusuff F, Ibrahim DSM, Mukhtar A, Joni AAM, Kusin FM, Mohamed KN, et al.

Mar Pollut Bull, 2021 Dec;173(Pt B):113071.
PMID: 34741924 DOI: 10.1016/j.marpolbul.2021.113071

This study aimed to identify the toxicity effect of Diuron on Tegillarca granosa by determining accumulation in the soft tissues and alteration on the gill tissue structure. Wild stocks T. granosa were grouped to identify the initial Diuron concentration and 48-h exposure in different concentrations (0, 1, 2, 3, and 4 mg·L-1). The Diuron in tissue was extracted using an established procedure and identified using High-Performance Liquid Chromatography (HPLC). Gills were analysed for histopathological study; tissue structure was classified and scored using a developed scoring system. Diuron concentration in the soft tissue of exposed T. granosa was in the range 1.27-8.09 mg·kg-1, w/w and not proportionately increase along with the exposure concentration. The mean index values of the gill histopathology ranged from 5.25-7.67 and classified as moderate to severe. Gills characteristics and T. granosa protective behaviour potentially limit Diuron accumulation in the soft tissue.
Relationship between Pb and Cd accumulations in house crow, their habitat, and food content from Klang area, Peninsular Malaysia

Janaydeh M, Ismail A, Omar H, Zulkifli SZ, Bejo MH, Aziz NAA

Environ Monit Assess, 2017 Dec 27;190(1):47.
PMID: 29282545 DOI: 10.1007/s10661-017-6416-2

Heavy metal pollution has become a global concern due to accumulation in tissue and transferable effects to humans via the food chain. This study focused on monitoring the accumulation of cadmium (Cd) and lead (Pb) in surface soil and body content: bone, heart, brain, liver, lung, muscle, kidney, feathers, feces, and gizzard contents of house crow Corvus splendens in the Klang region, Malaysia. The results revealed the occurrence of Pb and Cd in all biological samples from house crows, food contents, and surface soil samples. Heart and kidney accrued high amounts of Cd, while high amounts of Pb were found to accumulate in bones and feathers. Major discrepancies were also discovered in the concentrations of metals between juvenile and adults, as well as female and male bird samples. Concentrations of Pb and Cd in house crow internal tissues correlated significantly with that of bird feathers, but none could be established with that of surface soil. In addition, a significant correlation was observed between Pb concentration in the internal tissues to that of the feces, but the same was not the case when compared with the surface soil concentration. Metal accrual in the house crows feathers and feces may be through a long-term transmission via the food chain, which are eliminated from feathers via molting. This may suggest the utility of molted breast feathers of house crow in the bio-monitoring of Cd and Pb contamination, whereas feces of house crow appear only to be suitable for the bio-monitoring of Pb contamination.
Fulltext Economic contribution and attitude towards alien freshwater ornamental fishes of pet store owners in Klang Valley, Malaysia

Saba AO, Ismail A, Zulkifli SZ, Shohaimi S, Azmai MNA

PeerJ, 2021;9:e10643.
PMID: 33520451 DOI: 10.7717/peerj.10643

Malaysia is one of the top ten countries in the world that produce freshwater ornamental fishes. This industry can offer better livelihood opportunities to many poor households. However, most of the produced ornamental fishes are alien to Malaysia. In this study, we explore the contribution of alien freshwater fishes to the income of ornamental fish store owners and their attitude towards alien freshwater fishes within Klang Valley, Malaysia. Using a structured questionnaire, we surveyed 70 pet stores out of which 54 (81.42%) store owners responded. Most of the pet store owners were male (72%), Chinese (83%), and the highest educational level was at the secondary level (79%). Most of the pet store owners reported a monthly income of RM 2001-RM 5000 (78%) and were married (73%). Using Chi-square (χ2) test, significant relationships (p < 0.05) existed between the attitude of store owners towards alien ornamental fish species versus educational level (χ2 = 16.424, p = 0.007) and contribution of alien ornamental fishes to the pet store owners' income (χ2 = 27.266, p = 0.003). Fish sales as the main income source also related significantly with the impact of fish selling business on income level (χ2 = 10.448, p = 0.007). This study showed that the ornamental fish sales contributed over half of the income (51-100%) from the businesses of store owners. Almost half of the respondents (42%) reported that alien ornamental fish was the highest contributor to their income from the ornamental fish sale. While the mismanagement of alien ornamental fishes could give various negative ecological impacts, the socio-economic benefits of these fishes cannot be denied.
Fulltext Seasonal variation of heavy metals and metallothionein contents in Asian swamp eels, Monopterus albus (Zuiew, 1793) from Tumpat, Kelantan, Malaysia

Sow AY, Ismail A, Zulkifli SZ, Amal MN, Hambali K

BMC Pharmacol Toxicol, 2019 Jan 29;20(1):8.
PMID: 30696486 DOI: 10.1186/s40360-019-0286-x

BACKGROUND: Levels of toxic metal exposure in indigenous inhabitants are key bioindicators of the severity of environmental contamination. This study measured the seasonal variation of heavy metals and metallothionein (MT) contents in Asian swamp eels (Monopterus albus) from a paddy field situated in Tumpat, Kelantan, Malaysia, to identify prevalence, patterns and associations and togain insight on the suitability of MT as a biomarker for metal exposure.
METHODS: Gill, muscle and liver tissues of M. albus (n = 50) sampled during the ploughing, seedling, growing and harvesting phases of rice growing were collected. The concentrations of copper (Cu), zinc (Zn), lead (Pb), nickel (Ni), and cadmium (Cd) in these tissues were determined by flame atomic absorption spectrometry. MT from each sample was isolated and purified, and subsequently quantitated using UV spectrophotometry. Associations between metal and MT concentrations, season and tissue type were evaluated using Pearson correlation and ANOVA with post-hoc Tukey HSD analysis.
RESULTS: Zn was present in higher quantities in gill and liver tissues, while Cu levels were elevated solely in liver. Patterns of non-essential metal accumulation were varied: Cd was detected in low concentrations in all tissues, while Pb and Ni were abundant in gill tissues across all seasons. MT concentration in liver tissue was consistently higher than that found in muscle or gill tissue, except during the growing phase. Moreover, significant correlations (P
Fulltext Survey on Heavy Metals Contamination and Health Risk Assessment in Commercially Valuable Asian Swamp Eel, Monopterus albus from Kelantan, Malaysia

Sow AY, Ismail A, Zulkifli SZ, Amal MN, Hambali KA

Sci Rep, 2019 04 23;9(1):6391.
PMID: 31015502 DOI: 10.1038/s41598-019-42753-2

This work investigates the metals concentration in the tissues of Asian swamp eel, Monopterus albus. Five selected tissues, including liver, gill, bone, skin, and muscle were examined for the concentration of Zn, Cu, Cd, Pb, and Ni. The concentrations of Cd and Pb were found high in the muscle tissues of the eels. Additionally, high amounts of Zn and Cu metals were observed in the liver, whereas the Cd, Pb, and Ni metals were highly detected in gill. The accumulation of Zn, Cu, Cd, Pb, and Ni in both skin and bone of the eel seems to vary between seasons. Low levels of Zn, Cu, and Ni were identified in the muscle tissues of the eels. This study revealed that the concentration of Cd and Pb in the muscle tissues of Asian swamp eels exceeded the permissible limits by the US EPA, suggesting the consumption of the muscle may be hazardous and can severely affect one's health.
Assessment of P-gp and MRP1 activities using MultiDrugQuant Assay Kit: a preliminary study of correlation between protein expressions and its functional activities in newly diagnosed acute leukaemia patients

Fazlina N, Maha A, Zarina AL, Hamidah A, Zulkifli SZ, Cheong SK, et al.

Malays J Pathol, 2008 Dec;30(2):87-93.
PMID: 19291917

Multidrug resistance (MDR) is believed to be responsible for poor response of patients towards chemotherapy particularly patients with acute myeloid leukemia (AML) and acute lymphoblastic leukemia (ALL). The best-characterized resistance mechanism is the one mediated by permeability-glycoprotein (P-gp) encoded by MDR1 gene, which is responsible for drug efflux. We studied P-gp and multidrug resistance-associated protein 1 (MRP1) expression and functional activities in 43 newly diagnosed acute leukemia cases (19 paediatric ALL cases and 24 adult AML cases). The expression and functional activities were examined using flow cytometry and MultiDrugQuant assay kit (involving calcein AM uptake and efflux). P-gp and MRP1 expression and its functional activities were observed in 68.4% of paediatric ALL. In adult AML cases, all cases expressed MRP1 and its functional activities but only 58.3% were positive for P-gp and its functional activities. We were able to show a significant correlation between the expression of the multidrug resistant protein (P-gp and MRP1) and their functional activity in adult AML and paediatric ALL samples.
Expression of multidrug resistance (MDR) proteins and in vitro drug resistance in acute leukemias

Fazlina N, Maha A, Jamal R, Zarina AL, Cheong SK, Hamidah H, et al.

Hematology, 2007 Feb;12(1):33-7.
PMID: 17364990

The expression of the multidrug resistance (MDR) proteins may influence the outcome of treatment in patients with acute leukemia. The aim of this study was to determine the IC50 of cytotoxic drugs (cytosine arabinoside, ara-C and daunorubicin, dnr) using the in vitro 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)2H-tetrazolium, inner salt (MTS) assay method. A total of 82 newly diagnosed acute leukemia cases (43 adult myeloid leukaemia, AML cases and 39 acute lymphoblastic leukaemia, ALL cases) and 16 relapsed cases (8 AML cases and 8 ALL cases) were studied. The MTS assay was performed using two cytotoxic drugs, dnr and ara-C. Cells were incubated with different concentrations of drugs for 4 days and the IC50 was extrapolated from the viability curve. In newly diagnosed cases, we found that childhood ALL samples showed higher IC50 values of dnr (0.040 +/- 2.320) compared to adult AML samples (0.021 +/- 0.158). In contrast, newly diagnosed adult AML samples showed higher IC50 values of ara-C (0.157 +/- 0.529) compared to childhood ALL samples (0.100 +/- 2.350). In relapsed cases, two samples of childhood ALL showed IC50 values of dnr (0.910 +/- 1.760) and ara-C (1.310 +/- 2.390), which was higher compared to childhood AML samples (0.129 +/- 0.214 and 0.210 +/- 0.003, respectively). However, there was no correlation between IC50 values of these drugs tested with clinical outcome. In conclusion, we found that MTS assay is an easy, rapid and non laborious method to study in vitro drug resistance in acute leukaemia cases.
Evaluation of the status and distributions of heavy metal pollution in surface sediments of the Langat River Basin in Selangor Malaysia

Kadhum SA, Ishak MY, Zulkifli SZ, Hashim RB

Mar Pollut Bull, 2015 Dec 15;101(1):391-396.
PMID: 26476862 DOI: 10.1016/j.marpolbul.2015.10.012

The concentration of four metals: Cd, Ni, Cr, and Sn, in the surface sediment samples from the Langat River were evaluated. Multivariate techniques were used to apportion the sources of the metals. The results showed that the highest concentration of metals in the Langat River were found at Jenjarom station, with the concentration of these metals decreasing in the order of Sn>Cr>Ni>Cd (114.27, 21.03, 7.84, 0.59 μg g(-1) dry weight). The level of pollution in the sediment was assessed using contamination factor (CF), pollution load index (PLI), geo-accumulation index (Igeo), and enrichment factor (EF). The results of the pollution assessment showed that the Langat River sediments have severe enrichment of Sn and moderate to severe enrichment of Cd. The results of the PLI for the Langat River suggest that the sampling stations are not polluted with the exception of the Jugra, Jenjarom, and Jalan Hulu Langat stations.
Investigating geochemical factors affecting heavy metal bioaccessibility in surface sediment from Bernam River, Malaysia

Kadhum SA, Ishak MY, Zulkifli SZ, Hashim RB

Environ Sci Pollut Res Int, 2017 May;24(14):12991-13003.
PMID: 28378310 DOI: 10.1007/s11356-017-8833-8

The present study applied the use of sequential extraction technique and simple bioaccessibility extraction test to investigate the bioavailable fractions and the human bioaccessible concentration of metals collected from nine stations in surface sediment of the Bernam River. The concentrations of total and bioaccessible metals from different stations were in the range of 0.30-1.43 μg g(-1) and 0.04-0.14 μg g(-1) for total cadmium and bioaccessibility of cadmium, respectively, 6.20-288 μg kg(-1) and 2.06-8.53 μg kg(-1) for total mercury and bioaccessibility of mercury, respectively, and 9.2-106.59 μg g(-1) and 0.4-2.75 μg kg(-1) for total tin and bioaccessibility of tin, respectively. The chemical speciation of Cd in most sampling stations was in the order of oxidisable-organic > acid-reducible > residual > exchangeable, while the chemical speciation of Hg was in the order of exchangeable > residual > oxidisable-organic > acid-reducible and the chemical speciation of Hg was in the order of residual > oxidisable-organic > acid-reducible > exchangeable. The principal component analysis showed that the main factors influencing the bioaccessibility of mercury in surface sediments were the sediment total organic matter, cation exchange capacity, and easily, freely, or leachable and exchangeable fraction, and the factors influencing the bioaccessibility of tin were the total tin and cation exchange capacity, while the bioaccessibility of Cd in surface sediments was influenced by the only factor which is the easily, freely, or leachable and exchangeable fraction.
Fulltext Microplastics Pollution as an Invisible Potential Threat to Food Safety and Security, Policy Challenges and the Way Forward

Usman S, Abdull Razis AF, Shaari K, Amal MNA, Saad MZ, Mat Isa N, et al.

Int J Environ Res Public Health, 2020 Dec 21;17(24).
PMID: 33371479 DOI: 10.3390/ijerph17249591

Technological advances, coupled with increasing demands by consumers, have led to a drastic increase in plastic production. After serving their purposes, these plastics reach our water bodies as their destination and become ingested by aquatic organisms. This ubiquitous phenomenon has exposed humans to microplastics mostly through the consumption of sea food. This has led the World Health Organization (WHO) to make an urgent call for the assessment of environmental pollution due to microplastics and its effect on human health. This review summarizes studies between 1999 and 2020 in relation to microplastics in aquatic ecosystems and human food products, their potential toxic effects as elicited in animal studies, and policies on their use and disposal. There is a paucity of information on the toxicity mechanisms of microplastics in animal studies, and despite their documented presence in food products, no policy has been in place so far, to monitor and regulates microplastics in commercial foods meant for human consumption. Although there are policies and regulations with respect to plastics, these are only in a few countries and in most instances are not fully implemented due to socioeconomic reasons, so they do not address the problem across the entire life cycle of plastics from production to disposal. More animal research to elucidate pathways and early biomarkers of microplastic toxicity that can easily be detected in humans is needed. This is to create awareness and influence policies that will address this neglected threat to food safety and security.
Time dependent effect of chronic embryonic exposure to ethanol on zebrafish: Morphology, biochemical and anxiety alterations

Ramlan NF, Sata NSAM, Hassan SN, Bakar NA, Ahmad S, Zulkifli SZ, et al.

Behav Brain Res, 2017 08 14;332:40-49.
PMID: 28559182 DOI: 10.1016/j.bbr.2017.05.048

Exposure to ethanol during critical period of development can cause severe impairments in the central nervous system (CNS). This study was conducted to assess the neurotoxic effects of chronic embryonic exposure to ethanol in the zebrafish, taking into consideration the time dependent effect. Two types of exposure regimen were applied in this study. Withdrawal exposure group received daily exposure starting from gastrulation until hatching, while continuous exposure group received daily exposure from gastrulation until behavioural assessment at 6dpf (days post fertilization). Chronic embryonic exposure to ethanol decreased spontaneous tail coiling at 24hpf (hour post fertilization), heart rate at 48hpf and increased mortality rate at 72hpf. The number of apoptotic cells in the embryos treated with ethanol was significantly increased as compared to the control. We also measured the morphological abnormalities and the most prominent effects can be observed in the treated embryos exposed to 1.50% and 2.00%. The treated embryos showed shorter body length, larger egg yolk, smaller eye diameter and heart edema as compared to the control. Larvae received 0.75% continuous ethanol exposure exhibited decreased swimming activity and increased anxiety related behavior, while withdrawal ethanol exposure showed increased swimming activity and decreased anxiety related behavior as compared to the respective control. Biochemical analysis exhibited that ethanol exposure for both exposure regimens altered proteins, lipids, carbohydrates and nucleic acids of the zebrafish larvae. Our results indicated that time dependent effect of ethanol exposure during development could target the biochemical processes thus leading to induction of apoptosis and neurobehavioral deficits in the zebrafish larvae. Thus it raised our concern about the safe limit of alcohol consumption for pregnant mother especially during critical periods of vulnerability for developing nervous system.
Study on a new mechanism of sterilization in imposex affected females of tropical marine neogastropod, Thais sp

Mohamat-Yusuff F, Zulkifli SZ, Otake T, Harino H, Ismail A

J Environ Biol, 2014 Sep;35(5):995-1003.
PMID: 25204078

The morphological expressions and histopathological analysis of the gonads of a tropical marine neogastropod species (Thais sp.) from East Malaysia revealed new evidence of mechanical sterility in the imposex affected females. The gradual development of imposex was classified into five stages (Stage 0 to Stage 4) with three types of sterility conditions; Type A caused prohibition of copulation and capsule formation; Type B prohibits the releasing process of eggs; and gonads in Type C are infertile. Further analysis is needed to confirm, if the gonad malformation in imposex affected snails is generated specifically by tributyltin (TBT) or by other possible factors. The levels of imposex incidence (stages and percentages) were greater in a marina and decreased with increasing distance from the marina. Organotin tissue burden across the sexes showed that dibutyltin (DBT) as well as TBT might be the elements inducing imposex in Thais sp. from Miri in East Malaysia.
Notes on the occurrence of the tropical eel Anguilla bicolor bicolor in Peninsular Malaysia, Malaysia

Arai T, Chino N, Zulkifli SZ, Ismail A

J Fish Biol, 2012 Mar;80(3):692-7.
PMID: 22380562 DOI: 10.1111/j.1095-8649.2011.03154.x

Previous studies indicated that a tropical freshwater eel Anguilla bicolor bicolor occurs in Africa, India, Sri Lanka, Bangladesh, Myanmar, Indonesia and Australia, but an intensive survey has indicated an extended distribution range for the species into Peninsular Malaysia. Thus, A. b. bicolor is a native subspecies of Malaysia.
Fulltext Embryonic toxicity of 3,4-dichloroaniline (3,4-DCA) on Javanese medaka (Oryzias javanicus Bleeker, 1854)

Ibrahim MA, Zulkifli SZ, Azmai MNA, Mohamat-Yusuff F, Ismail A

Toxicol Rep, 2020;7:1039-1045.
PMID: 32913717 DOI: 10.1016/j.toxrep.2020.08.011

Early-life exposure to toxic chemicals causes irreversible morphological and physiological abnormalities that may last for a lifetime. The present study aimed to determine the toxicity effect of 3,4-Dichloroaniline (3,4-DCA) on Javanese medaka (Oryzias javanicus) embryos. Healthy embryos were exposed to various 3,4-DCA concentrations for acute toxicity (5, 10, 25, 50, and 100 mg.L-1) and sublethal toxicity (0.10, 0.50, 1.25, 2.50, and 5.00 mg.L-1) for 96 h and 20 days respectively. Acute toxicity test revealed that the median lethal concentration (96h-LC50) was 32.87 mg.L-1 (95 % CI = 27.90-38.74, R2 = 0.95). Sublethal exposure revealed that 1.25 mg.L-1 at 3 days post-exposure (3 dpe) has a significant lower heartrate (120 ± 12.3 beats/min., p
Fulltext Impact of Diuron contamination on blood cockles (Tegillarca granosa Linnaeus, 1758)

Mohamat-Yusuff F, Zulkarnain Z, Anuar NZA, Joni AAM, Kusin FM, Mohamed KN, et al.

Mar Pollut Bull, 2020 Dec;161(Pt A):111698.
PMID: 33022498 DOI: 10.1016/j.marpolbul.2020.111698

Examination of the impact of Diuron contamination on blood cockles (Tegillarca granosa) was conducted by combining field screening at three sampling events and a toxicity test. Diuron was extracted using the liquid-liquid extraction (LLE) technique and analyzed using HPLC-UV. The median lethal concentration (LC50) of Diuron on T. granosa was tested under a 72-h exposure. Diuron in water samples ranged from not detected (ND) to 3910 ppb, which was the highest concentration detected in samples after the irrigation water was discharged from the paddy plantation. Diuron was not detected in sediment samples. Mortality of T. granosa ranged from 4.74 to 38.33% with the highest percentages recorded after the release of the irrigation water. The LC50 value of Diuron was 1.84 ppm. This study suggests that irrigation water from paddy plantation that drifts to coastal areas containing Diuron harms T. granosa at the study area.
Fulltext Reproductive Toxicity of 3,4-dichloroaniline (3,4-DCA) on Javanese Medaka (Oryziasjavanicus, Bleeker 1854)

Ibrahim MA, Zulkifli SZ, Azmai MNA, Mohamat-Yusuff F, Ismail A

Animals (Basel), 2021 Mar 12;11(3).
PMID: 33809309 DOI: 10.3390/ani11030798

Compound 3,4-dichloroaniline (3,4-DCA) is a metabolite of several urea herbicides and intermediate chemical of several industrial products. Moreover, 3,4-DCA has been frequently detected in aquatic ecosystems around the world. This aniline is more toxic than the parent chemicals, and it affects non-target organisms. This study evaluated a 21-day reproductive response of an emerging aquatic vertebrate model, Javanese medaka (Oryzias javanicus), exposed to 3,4-DCA. Fecundity and gonads histopathology were observed. The spawning rate and fertilisation reduced significantly in the highest exposed-group (250 µg/L). Gonadosomatic index (GSI) was significantly low in females exposed to 250 µg/L. No substantial structural alteration of male gonads. However, oocyte development and ovarian cell structure were disrupted in 250 µg/L exposed females. The gonadal developmental was not affected in the males; however, a significant reduction in the developmental of female gonads was observed at 250 µg/L. These results show that 3,4-DCA interfere with the reproduction of Javanese medaka through fecundity and alteration of gonadal tissues.
Acute toxicity test of copper pyrithione on Javanese medaka and the behavioural stress symptoms

Mohamat-Yusuff F, Sarah-Nabila AG, Zulkifli SZ, Azmai MNA, Ibrahim WNW, Yusof S, et al.

Mar Pollut Bull, 2018 Feb;127:150-153.
PMID: 29475647 DOI: 10.1016/j.marpolbul.2017.11.046

This study was conducted to investigate the median lethal concentration (LC50) of copper pyrithione (CuPT) at 96-hr exposure on adult Javanese medaka (Oryzias javanicus) in revealing toxicological effects of CuPT contamination in the tropical area. Wild stock fishes were acclimatized for 14-days prior analysis. Triplicate of test tanks for seven test concentrations were placed with ten fishes each, this includes two control tanks. The behaviour of the tested fishes was manually observed through a camera. The LC50 of CuPT at 96-h was found to be 16.58mg/L. Tested fishes swam slowly in vertical movement and swam fast towards food during feeding time as the sign of stress behaviour. Meanwhile, fishes in the two control groups swam actively in a horizontal manner and no excitement during feeding time. No mortality in control groups. Results indicate CuPT to be toxic to Javanese medaka at low concentration and caused behavioural stress.

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