Metabolomics Study with Gas Chromatography-Mass Spectrometry of Hepatotoxicity on Adult Zebrafish (Brachydanio Rerio) after Exposure to Diquat

Although diquat is a widely used water-soluble herbicide in the world, its toxicity to freshwater fish has not been well characterized. In this study, gas chromatography-mass spectrometry (GC-MS) based metabolomics approach combined with histopathological examination and biochemical assays was applied to comprehensively assess the hepatotoxicity in zebrafish (Brachydanio rerio) after diquat exposure at two dosages of 0.34 and 1.69 mg·L−1 for 35 days. The results indicated that 1.69 mg·L−1 diquat exposure cause serious cellular swell and vacuolization with increased nuclear abnormality, and lead to obvious disturbance of antioxidative system and dysfunction in liver; while no obvious pathological injury could be detected, and changes in liver biochemistry were less pronounced at the dose level of 0.34 mg·L−1. Multivariate statistical analysis and pattern recognition showed different GC-MS profiles of zebrafish liver following exposure to diquat, the cluster of the treated groups were both clearly separated from the control samples. The differentially abundant metabolites mainly include carbohydrates, amino acids, lipids, nucleotides, and their derivatives. In the exposure group of 1.69 mg·L−1 diquat, severe disturbances of amino acid metabolism played important biological roles associated with inhibition of energy metabolism, reduced immunity, and disorders in neurotransmitters as pathway analysis revealed. Additionally, fluctuation of inositol, creatine, and pantothenic acid, substances associated with stress regulation and signal transduction, participating in metabolic abnormalities in zebrafish with diquat-triggered hepatic damage. Energy metabolism of zebrafish exposed on 0.34 mg·L−1 diquat more inclined to rely on anaerobic glycolysis than the normal ones. Amino acid metabolism responses were less affected, but obvious interference effects on lipid metabolism were observed with 0.34 mg·L−1 diquat exposure. These results imply increased sensitivity of metabolomics versus histopathology and clinical chemistry in recognizing liver toxicity of diquat. This study will contribute to explore possible mechanism of hepatic damages on nontarget freshwater fish induced by diquat and provide important basis for its environmental risk assessment.

Effect of Copper Ion Sterilization on Bacterial Community in a Freshwater Recirculating Aquaculture System

The study aimed to evaluate the safety of copper ion sterilization based on copper ion residues in zebrafish (Brachydanio rerio), as well as bacterial community structure and diversity in recirculating aquaculture systems (RASs). The copper ion content was determined using national food safety standard GB 5009.13-2017. Bacterial community structures and alpha and beta diversity indexes were examined using the 16S rRNA gene sequences produced by Illumina HiSeq sequencing. The results revealed no significant copper ion enrichment in B. rerio when the copper ion concentration was 0.15 mg/L. The relative abundances of Erythrobacter, nitrite bacteria, and Flavanobacteria were clearly higher in the treatment group than in the control and differences in bacterial species richness and diversity were obvious. In addition, there was no sharp decrease in the microflora at the outflow of the copper ion generator. In conjunction with the changes in ammonia nitrogen, nitrate, and nitrite concentrations during the experiment, the results indicated that there were no significant effects on the purification efficacy of the biological filter, but the abundances of beneficial bacteria increased significantly. This is of great relevance in order to understand the response of bacterial communities affected by changing environmental conditions, such as copper ion sterilization.

Fish community structure along altitudinal gradients with relation to environmental variables in Ratuwa River of Eastern, Nepal

Studies on fish community structure along altitudinal gradients of rivers are lacking in Nepal. This study was carried out to gauge the fish diversity and composition along elevational gradient in Ratuwa River. The altitudinal gradient varies between 70 m to 1300 m. Fish samples were collected based on habitat representativeness from April (spring) 10-18, July (summer) 10-18, October (autumn) 10-18, 2020 and January (winter) 10-18, 2021. A total of 3447 specimens representing 4 orders, 14 families and 36 species were identified. Both fish diversity and abundance of studied ichthyofauna vary with altitudinal gradient. The present study affirmed that fish species of Opsarius bendelisis, Schistura multifasciatus, Garra annandalei, Brachydanio rerio, Aspidoparia morar, and Schistura scaturigina are the major contributory species (>1%) for both space and time spectrums. Fish community structure testing for both to space and time showed significant difference in spatial spectrum (R=0.72, P<0.01) but no significant difference in temporal variation (R=-0.034, P>0.05). The present results hinted that fish assemblage structure varied significantly from low to high elevations, altitude, water temperature, water velocity, dissolved oxygen and pH was found as major influential factors (P<0.05) for species distribution.

Acute toxic effects of uranium on two aquatic organisms, Zebrafish (Brachydanio rerio) and Ostracod (Cypridopsis vidua)

In this paper, we concentrate on the acute toxic effects of uranium on two aquatic organisms, Brachydanio rerio (B. rerio) and Cypridopsis vidua (C. vidua). We found that the toxicity of uranium on C. vidua was significantly greater than that of B. rerio. The results show that C. vidua has a higher sensitivity to uranium, even better than the commonly used test model organisms. In addition to its high sensitivity, C. vidua is a simple and cost-effective toxicological test organism. Therefore, C. vidua is an attractive alternative biological detection material. The acute toxicity results of the test are valuable for establishing water quality standards and protecting human health. At the same time, it enriches the relevant data of uranium on biological toxicity, provides clues for the study of the mechanism of toxicity, and deepens the understanding of the harm of uranium pollution to aquatic ecosystems.

Review on BOD/COD Ratio Toxicity to Daphnia magna, Artemia salina and Brachydanio rerio

This paper review research works on BOD/COD ratio toxicity to three bioindicators, namely: Daphnia magna, Artemia salina, and Brachydanio rerio. Treatment methods are divided into natural, biological, physical, chemical, combined treatment, and phytotechnology for various types of processing operations such as municipal and industrial wastewater treatment plants, various effluent treatment, landfill leachate, and organic matter. A. salina shows the lowest toxicity value of the BOD/COD ratio and shows it can withstand processing conditions that are not biodegradable. Then followed by B. rerio and D. magna. Furthermore, the disposal limits are standardized for the protection of freshwater biota. In addition, it must be considered whether there is any other potential of disposal into the receiving environment, and if so, what form of disposal and how much, to protect the biota.

SDHI Fungicide Toxicity and Associated Adverse Outcome Pathways: What Can Zebrafish Tell Us?

Succinate dehydrogenase inhibitor (SDHI) fungicides are increasingly used in agriculture to combat molds and fungi, two major threats to both food supply and public health. However, the essential requirement for the succinate dehydrogenase (SDH) complex—the molecular target of SDHIs—in energy metabolism for almost all extant eukaryotes and the lack of species specificity of these fungicides raise concerns about their toxicity toward off-target organisms and, more generally, toward the environment. Herein we review the current knowledge on the toxicity toward zebrafish (Brachydanio rerio) of nine commonly used SDHI fungicides: bixafen, boscalid, fluxapyroxad, flutolanil, isoflucypram, isopyrazam, penthiopyrad, sedaxane, and thifluzamide. The results indicate that these SDHIs cause multiple adverse effects in embryos, larvae/juveniles, and/or adults, sometimes at developmentally relevant concentrations. Adverse effects include developmental toxicity, cardiovascular abnormalities, liver and kidney damage, oxidative stress, energy deficits, changes in metabolism, microcephaly, axon growth defects, apoptosis, and transcriptome changes, suggesting that glycometabolism deficit, oxidative stress, and apoptosis are critical in the toxicity of most of these SDHIs. However, other adverse outcome pathways, possibly involving unsuspected molecular targets, are also suggested. Lastly, we note that because of their recent arrival on the market, the number of studies addressing the toxicity of these compounds is still scant, emphasizing the need to further investigate the toxicity of all SDHIs currently used and to identify their adverse effects and associated modes of action, both alone and in combination with other pesticides.

Toxic Effects of Heavy Metals and Organic Polycyclic Aromatic Hydrocarbons in Sediment Porewater on the Amphipod Hyalella azteca and Zebrafish Brachydanio rerio Embryos from Different Rivers in Taiwan

The amphipod (Hyalella azteca) and zebrafish (Brachydanio rerio) embryos were used for toxicological sediment porewater testing. Porewaters from 35 sampling stations of eight streams in southern Taiwan were screened for toxic effects and their relationship with 6 metal and 16 polycyclic aromatic hydrocarbons (PAHs). Concentration analysis results showed that the following PAHs, naphththalene, benzo(b)fluoranthene, dibenz(a,h)anthracene, acenaphthalene, and the heavy metal cadmium were not detected in 35 sampling stations. The highest detection rate of 94.1% was caused by the PAHs fluoranthene and pyrene. The highest detection rate of the metal zinc was 88.6% of 35 analyzed samples. The majority of samples (88%) were classified as level tier 1 according to USEPA national sediment inventory. This indicates the probability of adverse effects on aquatic life or human health. The results of a zebrafish embryo test showed that heart rate and survival were significantly reduced with all porewater samples. Therefore, fish exposed to contaminated river conditions may be affected in their cardiovascular functions. Looking at correlations between toxic effects of metals and PAHs, we found that phenanthrene, anthracene, pyrene, benzo(a)anthracene, chrysene, benzo(b)fluoranthene, and benzo(a)pyrene were low, while fluorene was highly correlated with toxic effects of metals.

Review: Zebrafish (Danio Rerio) Sebagai Model Obesitas dan Diabetes Melitus Tipe 2

Zebrafish (Danio rerio) yang sebelumnya disebut Brachydanio rerio merupakan spesies ikan air tawar tropis yang berukuran kecil yang berasal dari Asia Selatan dan Sungai Gangga di India. Zebrafish bisa diaplikasikan pada model obesitas dan diabetes melitus tipe 2 (DMT2). Obesitas merupakan pemicu terjadinya DMT2. Tujuan dari review artikel ini dapat memberikan informasi dan meningkatkan pemahaman pembaca bahwa zebrafish merupakan model yang ideal untuk obesitas dan DMT2. Metode penulisan review artikel ini dilakukan dengan penelusuran artikel ilmiah terpublikasi bertaraf nasional dan internasional dengan menggunakan kata kunci berupa “Obesity, zebrafish, model”, “Hyperglycemia, zebrafish, model” atau “zebrafish for hyperglycemia” dan “Diabetes melitus type 2, zebrafish, model”. Zebrafish model obesitas dan DMT2 yang diinduksi makanan dan obat menunjukkan keunggulan menjadi obesitas seperti pada manusia dan mamalia lain. Selain itu, zebrafish yang diinduksi dengan metode perendaman glukosa diperoleh hasil meningkatnya kadar glukosa darah dan gangguan respon terhadap insulin eksogen. Pemberian alternatif terapi dari bahan alam seperti GTE, palmaria mollis, diosgenin, bubuk kayu manis pada zebrafish memberikan hasil yang cukup baik sehingga bisa dipertimbangkan sebagai alternatif model hewan lain.

Fish Diversity and Water Quality Parameters of Mechi River, Jhapa, Province No. 1, Nepal

Species diversity is widely recognised as an important trait of functioning and resilient ecosystem. Spatio-temporal patterns of fish community structure in the Mechi River was studied based on stratified random survey, conducted between September 2018 and April 2019. The survey was conducted at three different sampling stations in three different seasons. For the fish sampling, two cast nets of different sizes were used, one having large mesh size of 0.5 cm, 3 m diameter and 2 kg weight and another having mesh size of 2 cm, 6 m diameter and 6 kg weight. In addition, drag net and gill net were also used to collect the fish samples. A total of 1,772 fishes belonging to 4 orders, 8 families, 16 genera and 33 species were documented. The results from the similarity percentage analysis indicated that, the species Schistura devdevi, Brachydanio rerio, Puntius sophore, Barilius barila, Schistura beveani and Puntius terio were the most copious fish species recorded from the Mechi River. One-way analysis of similarity testing for space and time variations in fish community indicated that there was a significant difference in space (R = 0.66, p<0.01) but no significant difference in time (R = -0.0123, p>0.433). The Canonical Correspondence Analysis hinted that dissolved oxygen, water velocity and pH have shown to influence the fish assemblage structure in Mechi River.

Dependence of Biocatalysis on D/H Ratio: Possible Fundamental Differences for High-Level Biological Taxons

The kinetics of biological reactions depends on the deuterium/protium (D/H) ratio in water. In this work, we describe the kinetic model of biocatalytic reactions in living organisms depending on the D/H ratio. We show that a change in the lifetime or other characteristics of the vital activity of some organisms in response to a decrease or increase in the content of deuterium in the environment can be a sign of a difference in taxons. For animals—this is a curve with saturation according to the Gauss’s principle, for plants—it is the Poisson dependence, for bacteria a weakly saturated curve with a slight reaction to the deuterium/protium ratio toward increasing deuterium. The biological activity of the aquatic environment with reduced, elevated, and natural concentrations of deuterium is considered. The results of the study are presented in different vital indicators of some taxons: the bacteria kingdom—the colony forming units (CFU) index (Escherichia coli); animals—the activation energy of the death of ciliates (Spirostomum ambiguum), embryogenesis of fish (Brachydanio rerio); plants—germination and accumulation of trace elements Callisia fragrans L., sprouting of gametophores and peptidomics of moss Physcomitrella patens. It was found that many organisms change their metabolism and activity, responding to both high and low concentrations of deuterium in water.