Journal of Soil Science and Plant Physiology (2025)

Changes in water quality have an adverse effect on organisms when they are subjected to conditions and high concentrations that result in their death [1]. This may be due to the usage of herbicides in business and agriculture. In an attempt to increase agricultural output, humans utilize heavy pesticides to protect crops from rodents and insects, beginning with pre-planting and continuing through weed control, crop cultivation, and storage. These compounds are toxic to humans, fish, and other aquatic life [2]. Any alteration in fish behavior sheds light on behavioral shifts that could be linked to physiological markers in aquatic species [3]. Because behavioural bioassays are quicker, more sensitive, and more relevant to the environment, they are frequently utilized in toxicity assessments [4]. Pollutant concentrations in water bodies have the potential to significantly increase aquatic habitat mortality. On the other hand, low concentrations cause the contaminants to bioaccumulate and biomagnify, which ultimately reaches humans through the food web. To guarantee that we prioritize eating healthful fish, the problem of water contamination needs to be taken very seriously and addressed with the utmost care [5].

While the use of chemicals in farming has many benefits, there are also big disadvantages, like pollution and environmental degradation, which are directly linked to the use of chemicals [6]. Agrochemical contamination poses a serious threat to environmental safety, and exposure to these chemicals can have negative health effects like cancer and neurological damage [7]. Because agricultural chemicals are dangerous for the ecosystem and all living things, and because they can easily contaminate water bodies, causing extensive harm to non-target species, including fish, their presence in ecosystems has become a major source of social and scientific anxiety worldwide [8]. Globally, agrochemical pollution of the environment has become a serious threat to wildlife conservation and human health [9]. Chemicals that are sprayed directly to aquatic systems, spray drifts, dust and rain fallout, sewage, industrial effluent, and infrequently, accidents, can all contaminate water, whether on purpose or by mistake.

In contrast to other pollutants, pesticides are intentionally released into the environment with the intention of employing their toxic properties to reduce disease-carrying insects and other pests. 99.9% of pesticides sprayed on targeted targets wind up in various environmental media, whereas less than 0.1% actually reach the pests [9]. Pesticides in streams gradually vanish due to torrents, dilution, partitioning in water or air, attachment to sediment particles, accumulation in aquatic creatures' tissues, or burial in sediment [10]. Surface runoff from intermittent rainfall is the primary way that pesticides are transferred from land to aquatic bodies, exposing non-target organisms like fish to pesticides on a pulse basis. Pesticide dissipation may be the cause of pulse exposure in lotic (flowing) systems, whereas hydrology (constant replenishment by water movement) may be the cause in lentic (still) systems [11]. Floods, dilution, partitioning in water or air, attachment to sediment particles, accumulation in aquatic animal tissues, or burial in the sediment can all result in pulse exposure to low pesticide concentrations [12].

Biomarkers are sub-organism responses in organisms that can demonstrate exposure to or the effects of environmental pollutants [13]. Oxidative damage, haematological changes, biochemical and histological abnormalities, genotoxicity, and mutagenicity are some of the indicators that are commonly evaluated in fish. Biomarkers can be used as early warning systems for environmental hazards and to monitor fish health [14]. Despite more than twenty years of research on aquatic organisms exposed to pesticides or other toxicants, nothing is known about the metabolic reactions of S. melanotheron to these substances. These creatures have a significant impact on the economy and ecology. The influence of atrazine on S.melanotheron metabolites is not well documented in the literature. Therefore, this work assessed S. meleanotheron's metabolic responses to different lab-based Adhatoda bucchulzii plant extracts doses.

Experimental Location and Fish

The study was carried out at the Nigerian Institute for Oceanography and Marine Research's branch office, the African Regional Aquaculture Center, located in Buguma, Rivers State, Nigeria. The 180 S. melanotheron utilized in the experiment were obtained from the center's recruiting ponds at low tide. After that, they were divided into three groups according to their sizes: juveniles (mean length 13.55cm±1.56SD and mean weight 68.99g±3.77SD) made up Group 1; sub-adults (mean length 15.32cm±3.45SD and mean weight 110.55g±12.87SD) made up Group 2; and adults (mean length 19.88cm±6.54SD and mean weight 148.76g±12.86SD) made up Group 3. Six open 50-liter plastic containers were used to transport the fish to the lab, where they were allowed to acclimate for seven days.

Preparation of Test Solutions and Exposure of Fish

Adhatoda bucchulzii plant extracts was used in the current investigation. The plant was collected from a forest in Port Harcourt. The leaves of the plant was rinsed and macerated using a blender machine. The plant extracts was obtained by adding 10.0 litre of water to 1kg of mashed plant. The extracts was administered to S.melanotheron in triplicate at dosages of 0.00 (control), 0.05, 0.10, 0.15, and 0.20 mg/L. Each test tank had 10 fish that were chosen at random. The duration of the experiment was fifteen days. Every day, fresh water was added to the tanks. The fish were fed twice daily at 3% body weight with a commercial feed.

Determination of Blood Plasma Metabolites

A 2ml sample of fresh blood was taken at the conclusion of each experimental period by puncturing the caudal artery with a tiny needle and pouring the sample into heparinized sample vials. Serum was separated by centrifugation in a TG20-WS Tabletop High Speed Laboratory Centrifuge for 5-8 minutes at 10,000 rpm. Following the guidelines provided by APHA [15], the samples were examined for the metabolites creatinine, total bilirubin, total urea, and total protein. There were three copies of each test run. The methods APHA [20] were also used to determine water quality parameters.

Statistical Analysis

The mean and standard deviation of the mean were used to express all the data. The data analysis was done using SPSS Version 22, a statistical program. Using two-way ANOVA, the means were split, and the two means were deemed significant at 5% (P <0.05).

The water quality parameters (Table 1) were all within the same range, except for DO, which showed lower values at higher chemical concentrations. Table 2 presents the effects of the Adhatoda bucchulzii plant extracts on the metabolites in the plasma of juvenile S.melanotheron. It was observed that the levels of creatinine, total protein, and total bilirubin decreased with increasing Adhatoda bucchulzii plant extracts concentrations, while the levels of urea significantly increased when compared to the control values. Table 3 shows the effects of the chemical atrazine on the metabolites in the plasma of sub-adult sizes of S.melanotheron. It was observed that the levels of creatinine, total protein, and total bilirubin decreased with increasing Adhatoda bucchulzii plant extracts concentrations, while the levels of urea significantly increased when compared to the control values.

Table 1: Physico-chemical Parameters of Water in Experimental Tanks (Meant± SD).

Means within the row with different superscripts are significantly different (P<0.05).

Table 2: Metabolite Activities in Juveniles of S. melanotheron Exposed to Adhatoda bucchulzii Plant Extracts.

Means within the same column with different super scripts are significantly different (P<0.05).

Table 3: Metabolite Activities in Sub-Adults of S. melanotheron Exposed to Adhatoda bucchulzii Plant Extracts.

Means within the same column with different super scripts are significantly different (P<0.05).

Table4: Metabolite Activities in Adults of S. melanotheron Exposed to Adhatoda bucchulzii Plant Extracts.

Means within the same column with different super scripts are significantly different (P<0.05).

Generally speaking, metabolite alterations can result from the impact of water quality features, especially in hazardous water. In this study, the water used in the experiment had no bearing on the metabolites' alterations and variations after the chemical is administered. This is because the different amounts of the solution did not significantly differ from the control in terms of temperature, salinity, pH, ammonia, or nitrite (P>0.05). The dissolved oxygen measurements, however, revealed lower concentrations. The pattern in the water quality indicators is similar to what other authors who treated fish to varying chemical concentrations in a salinity chamber found [16]. The main purpose of blood or plasma is to transport ingested metabolites and nutrients (both organic and inorganic) throughout the body, as well as waste materials to various excretory organs for their eventual removal [17].The existence of metabolites in the blood, whether in high or low amounts, is an important clinical correlation. The total bilirubin measurements dropped as the concentration of the chemical solution increased. A decrease in plasma total bilirubin indicates damage to the liver cells [23]. The investigation's lower bilirubin levels raise the possibility that S.melanotheron's liver cells were harmed. A decrease in total bilirubin concentrations in the plasma may also be due to the liver's incapacity to convert bilirubin into bile and urobilin, which is the condition that gives human urine its yellow hue [18].

Urea and creatinine have been used as important indicators for evaluating the impact of stress on the kidney using a variety of in-vivo and in-vitro methods [19]. Over the course of the exposure period, the experimental fish's urea levels increased as the concentration of the chemical solution increased, while its creatinine decreased. According to Calbreath [20], the kidney's inability to get rid of these waste products was demonstrated by a drop in glomerular filtration rate and an increase in urea content. Total protein content is used as a basic sign of fish health and is an important non-specific immunological marker [21]. Serum protein levels in the experimental fishes dropped in the current study. Total protein levels can drop as a result of reduced amino transferase activity, changed liver structure, and inadequate fluid balance management. This result confirms the findings of Anyanwu et al. [21] regarding the exposure of Sarotherodon melanotheron to varying salinities. The decrease in protein content was most likely caused by decreased or disrupted microsomal protein synthesis. Proteolytic activity may rise as a result of protein breakdown, and the outcome may be used for metabolic processes [22]. While a decrease in protein synthesis and an increase in proteolysis may occasionally result in a loss in protein content, an increase in protein synthesis may be associated with an increase in protein content due to an increase in the enzyme activity involved in protein synthesis [23]. However, following stress, scientists have seen a decrease in fish protein content.

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