Pharmacokinetics of praziquantel and its metabolites in grass carp (Ctenopharyngodon idella) following the oral administration of a single bolus.

The study aimed to evaluate and compare the routes and rates of the depletion of the antiparasitic praziquantel (PZQ), a derivative of pyrazinoisoquinoline, following its oral administration in grass carp (Ctenopharyngodon idella). We focused on the depletion of PZQ and its major metabolites - cis-hydroxy praziquantel (CPZQ) and trans-hydroxy praziquantel (TPZQ), in water, the plasma, hepatopancreas, kidney, muscle, and skin, following a single oral administration of PZQ in a concentration of 50 mg/kg. Fish were sampled before the drug administration and then eight times in the course of the 30-day-long experiment. Our results indicate the rapid absorption and elimination of PZQ and its metabolites in all the analysed matrices. The most PZQ-burdened tissue was the hepatopancreas, the gill and the skin. In all the samples, the concentration of the drug and its metabolites consistently declined over time. The residue of the parent compound was detected for the longest time in all the tissues. During the study, a significant (P < 0.01) correlation was found within the concentration of PZQ, CPZQ, and TPZQ in the water and all the biological matrices. It was also found that the PZQ residue was not detected below the maximum residue levels (i.e., 20 µg/kg) until 16 days after exposure in the muscle and skin.

In-feed Praziquantel treatment of grass carp (Ctenopharyngodon idella) infected with eye flukes.

Praziquantel (PZQ) is a drug effective against several parasitic diseases of fish caused by Platyhelminthes. The study assesses the efficacy of selected in-feed PZQ doses in eliminating metacercariae Diplostomum pseudospathaceum in grass carp. The fish were infected with D. pseudospathaceum in laboratory conditions and subsequently treated with PZQ at daily doses of 30 (PZQ1), 50 (PZQ2), and 100 (PZQ3) mg/kg bw for seven days. The treatment trial was followed by a 23-day observation period during which the fish were parasitologically examined on days 0, 2, 8 and 23. Smaller fish were infected with a significantly higher (P < 0.05) number of metacercariae than bigger fish among three weight groups as follows: up to 7 g > 7 to 12 g > 12 g. The in-feed PZQ at doses of 30, 50 and 100 mg/kg bw for seven days was 47%, 84% and 88% effective against metacercariae D. pseudospathaceum, respectively. A significant decrease (P < 0.01) in moving and an increase (P < 0.01) in the immotile metacercariae in the eye's lens were recorded in all the trial groups on each sampling day. An increasing proportion of motile metacercariae in the post-treatment observation period was seen in PZQ2 and PZQ3, indicating a decreasing trend in the actual efficacy in an environment with no reinfection possibility.

Trace elements in the muscle and liver tissues of Garra shamal from the freshwater ecosystem of Oman: an exposure risk assessment.

Anthropogenic activities lead to environmental contamination with foreign substances such as heavy metals. This work was aimed to monitor trace elements (total arsenic (As), cadmium (Cd), chrome (Cr), cobalt (Co), copper (Cu), lead (Pb), manganese (Mn), mercury (Hg), nickel (Ni), and zinc (Zn)) contamination levels (dry weight base) in three natural freshwater reservoirs of Oman including Al Khawd and Al Amarat (Muscat Governorate) and Surur area (Ad Dakhiliyah Governorate as control area) using a native benthic inland fish (Garra shamal; Cyprinidae) for the first time. The muscle and liver of a hundred and twenty G. shamal were collected to assess the degree of metal contamination. Atomic absorption spectrometry was used as an analytical technique. From the spectrum of analyzed elements, we found Zn as a major element in monitored areas. The statistically significant (P < 0.05) highest concentrations of Zn liver (0.275 ± 0.065 µg/g) were in Al Amarat compared to the other areas. The concentrations of monitored elements in the fish muscle were lower than the liver samples. Furthermore, the fish length was significantly correlated with the accumulation of Hg and Co in both muscle and liver samples. In all analyzed fish from Oman inland water, the concentrations of elements were below the permissible limits; however, additional research is needed.

Runoff of foliar-applied natural fungicides in aquaponics: Implications for fish and nitrification.

Aquaponics is a method of producing food in a sustainable manner through the integration of aquaculture and hydroponics, which allows simultaneous cultivation of fish and economic crops. The use of natural fungicides are crucial to the sustainable control of diseases in aquaponics. We assessed the potential impacts of natural fungicides, such as clove oil and lecithin, as well as a synthetic fungicide, tebuconazole, following foliar application in aquaponics. This study examined the runoff rates of the fungicides in decoupled aquaponics, and the subsequent effects of the runoffs on nitrification processes and Nile tilapia (Oreochromis niloticus). The runoffs of the foliar-applied fungicides, clove oil, lecithin, and tebuconazole, were detected in aquaponics water at a percentage runoff rate of 0.3 %, 2.3 %, and 0.3-0.8 % respectively. In the biofilter, lecithin altered the ammonium levels by increasing ammonium-nitrogen levels by 7 mg L-1, 6 h post application. Clove oil, on the other hand, showed no significant effect on ammonium, nitrite, and nitrate-nitrogen. Similarly, the toxicity test showed that eugenol had no significant effects on the hematological, biochemical and antioxidative activities of O. niloticus. Conversely, tebuconazole exhibited significant and persistent effects on various biochemical parameters, including lactate, albumin, and total protein, as well as hematological parameters like hemoglobin and MCH. The use of lecithin and tebuconazole should only be limited to decoupled aquaponics.

Selected neonicotinoids and associated risk for aquatic organisms.

Neonicotinoids are one of the newest groups of systemic pesticides, effective on a wide range of invertebrate pests. The success of neonicotinoids can be assessed according to the amount used, for example, in the Czech Republic, which now accounts for 1/3 of the insecticide market. The European Union (EU) has a relatively interesting attitude towards neonicotinoids. Three neonicotinoid substances (imidacloprid, clothianidin and thiamethoxam) were severely restricted in 2013. In 2019, imidacloprid and clothianidin were banned, while thiamethoxam and thiacloprid were banned in 2020. In 2022, another substance, sulfoxaflor, was banned. Therefore, only two neonicotinoid substances (acetamiprid and flupyradifurone) are approved for outdoor use in the EU. Neonicotinoids enter aquatic ecosystems in many ways. In European rivers, neonicotinoids usually occur in nanograms per litre. Due to the low toxicity of neonicotinoids to standard test species, they were not expected to significantly impact the aquatic ecosystem until later studies showed that aquatic invertebrates, especially insects, are much more sensitive to neonicotinoids. In addition to the lethal effects, many studies point to sublethal impacts - reduced reproductive capacity, initiation of downstream drift of organisms, reduced ability to eat, or a change in feeding strategies. Neonicotinoids can affect individuals, populations, and entire ecosystems.

Fitness assessment of Mytilus galloprovincialis Lamarck, 1819 after exposure to herbicide metabolite propachlor ESA.

The lack of data on the chronic effects of chloroacetanilide herbicide metabolites on non-target aquatic organisms creates a gap in knowledge about the comprehensive impacts of excessive and repeated pesticide use. Therefore, this study evaluates the long-term effects of propachlor ethanolic sulfonic acid (PROP-ESA) after 10 (T1) and 20 (T2) days at the environmental level of 3.5 µg.L-1 (E1) and its 10x fold multiply 35 µg.L-1 (E2) on a model organism Mytilus galloprovincialis. To this end, the effects of PROP-ESA usually showed a time- and dose-dependent trend, especially in its amount in soft mussel tissue. The bioconcentration factor increased from T1 to T2 in both exposure groups - from 2.12 to 5.30 in E1 and 2.32 to 5.48 in E2. Biochemical haemolymph profile and haemocyte viability were not affected by PROP-ESA exposure. In addition, the viability of digestive gland (DG) cells decreased only in E2 compared to control and E1 after T1. Moreover, malondialdehyde levels increased in E2 after T1 in gills, and DG, superoxidase dismutase activity and oxidatively modified proteins were not affected by PROP-ESA. Histopathological observation showed several damages to gills (e.g., increased vacuolation, over-production of mucus, loss of cilia) and DG (e.g., growing haemocyte trend infiltrations, alterations of tubules). This study revealed a potential risk of chloroacetanilide herbicide, propachlor, via its primary metabolite in the Bivalve bioindicator species M. galloprovincialis. Furthermore, considering the possibility of the biomagnification effect, the most prominent threat poses the ability of PROP-ESA to be accumulated in edible mussel tissues. Therefore, future research about the toxicity of pesticide metabolites alone or their mixtures is needed to gain comprehensive results about their impacts on living non-target organisms.

Responses of signal crayfish Pacifastacus leniusculus to single short-term pulse exposure of pesticides at environmentally relevant concentrations.

Although pesticides are often discharged into surface waters in pulses as opposed to a sustained release, the effect of episodic pollution events on freshwater crayfish is largely unknown. We monitored change in heart rate and distance moved to assess the response of signal crayfish Pacifastacus leniusculus to short-term exposure to environmentally relevant concentrations of metazachlor (MTZ), terbuthylazine (TER), and thiacloprid (TCL). Crayfish exposed to 20 µg/L of MTZ exhibited a significant increase in mean heart rate and distance moved. Increased heart rate was detected at 118 ± 74 s post-exposure to MTZ. There were no significant differences in mean heart rate and distance moved in crayfish exposed to 6 µg/L of TCL and 4 µg/L of TER. A significant correlation between heart rate and distance moved was found in all exposed groups. These results suggest that pulse exposure to MTZ impact crayfish physiology and behavior during short-term period. With pulse exposure to TCL and TER, crayfish not exhibiting a locomotor response may continue to be exposed to lower, but potentially harmful, levels of pollutants. Evidence of the impacts of pesticide pulse at environmentally relevant concentrations on crayfish is scarce. Further study is required to determine the ecological effects of such events on freshwater crayfish.

Multi-characteristic toxicity of enantioselective chiral fungicide tebuconazole to a model organism Mediterranean mussel Mytilus galloprovincialis Lamarck, 1819 (Bivalve: Mytilidae).

The survey of available scientific literature shows a lack of data on the chronic effects of tebuconazole (TEB) on non-target aquatic organisms. Therefore, this study evaluates toxicity (10 and 20 days) of two considered concentrations 2 ng/L (E1) and 2 µg/L (E2) of TEB to bioindicator species Mytilus galloprovincialis. To this end, the TEB concentrations measured in soft mussel tissues showed a time-dependent increasing trend. The viability of haemocyte and digestive gland (DG) cells was higher than 95 % during the experiment. However, DG cells lost the ability to regulate their volume in both groups after 20-d. The E1 treatment increased Cl- and Na+ levels, and E2 decreased Na+ levels in the haemolymph. In addition, levels of superoxide dismutase (SOD) activity and oxidatively modified protein (OMP) increased after 10- and 20-d in both treatments. Histopathological findings showed abnormalities in the E2, e.g., haemocyte infiltration, hypertrophy, and hyperplasia in gills and DG. This study reveals the potential risks of TEB usage in the model organism M. galloprovincialis, primarily via bioaccumulation of TEB in food web links, and improves knowledge about its comprehensive toxicity.

An update about beneficial effects of medicinal plants in aquaculture: A review.

Aquaculture is an essential and growing component of agricultural and global ecosystems worldwide. Aquaculture provides more than 25% of the total aquatic food consumption by humans. The development of the aquaculture industry should be followed in successive industrial years, and therefore it is necessary to pay attention to the management and type of farming system that is compatible with the environment. The use of antibiotics for disease control has been criticised for their negative effects, including the emergence of antibiotic-resistant bacteria, the suppression of the immune system and the environment, and the accumulation of residue in aquatic tissues. The use of these products reduces the need for treatments, enhances the effect of vaccines, and, in turn, improves production indicators. Medicinal plants have increasingly been used in recent years as a disease control strategy in aquaculture, boosting the immune system of aquatic animals and helping to develop strong resistance to a wide range of pathogens. Therefore, this review aims to provide an overview of the recent evidence on the beneficial use of medicinal plants to promote growth and strengthen the immune system in farmed aquatic animals.

Effects of praziquantel on common carp embryos and larvae.

This study aimed to assess the toxicity of praziquantel (anthelmintic drug) in different developmental stages of common carp (Cyprinus carpio) based on mortality, early ontogeny, growth, oxidative stress, antioxidant enzymes, histology and behaviour. Praziquantel at all tested concentrations ranging from 1 to 4 mg/L showed no significant adverse effects on mortality, the early ontogeny and behaviour locomotory (activity, moved distance and velocity) of carp after 35-day exposure. Concentrations of 3 and 4 mg/L caused significantly (P < 0.01) lower growth, total superoxide dismutase and catalase activities compared with controls. Praziquantel is safe for the early life of carp in concentrations ≤ 2 mg/L.

Integrated biomarker response in signal crayfish Pacifastacus leniusculus exposed to diphenhydramine.

Diphenhydramine (DPH) is a pharmaceutical with multiple modes of action, primarily designed as an antihistamine therapeutic drug. Among antihistamines, DPH is a significant contaminant in the environment, frequently detected in surface waters, sediments, and tissues of aquatic biota. In the present study, signal crayfish Pacifastacus leniusculus was used as a model organism because of their prominent ecological roles in freshwater ecosystems. The biochemical effects were investigated in crayfish exposed to the environmental (low: 2 µg L-1), ten times elevated (medium: 20 µg L-1), and the sublethal (high: 200 µg L-1) nominal concentrations of DPH in water for 96 h. Lipid peroxidation, antioxidant enzyme activities, and acetylcholinesterase activity were assessed as toxicological biomarkers in crayfish hepatopancreas, gills, and muscles. Low and medium DPH exposure caused imbalances only in glutathione-like enzyme activities. Integrated biomarker response showed the absolute DPH toxicity effects on all tested tissues under high exposure. This study identified that high, short-term DPH exposure induced oxidative stress in crayfish on multiple tissue levels, with the most considerable extent in muscles.

Chronic Toxicity of Primary Metabolites of Chloroacetamide and Glyphosate to Early Life Stages of Marbled Crayfish Procambarus virginalis

Degradation products of herbicides, alone and in combination, may affect non-target aquatic organisms via leaching or runoff from the soil. The effects of 50-day exposure of primary metabolites of chloroacetamide herbicide, acetochlor ESA (AE; 4 µg/L), and glyphosate, aminomethylphosphonic acid (AMPA; 4 µg/L), and their combination (AMPA + AE; 4 + 4 µg/L) on mortality, growth, oxidative stress, antioxidant response, behaviour, and gill histology of early life stages of marbled crayfish (Procambarus virginalis) were investigated. While no treatment effects were observed on cumulative mortality or early ontogeny, growth was significantly lower in all exposed groups compared with the control group. Significant superoxide dismutase activity was observed in exposure groups, and significantly higher glutathione S-transferase activity only in the AMPA + AE group. The gill epithelium in AMPA + AE-exposed crayfish showed swelling as well as numerous unidentified fragments in interlamellar space. Velocity and distance moved in crayfish exposed to metabolites did not differ from controls, but increased activity was observed in the AMPA and AE groups. The study reveals the potential risks of glyphosate and acetochlor herbicide usage through their primary metabolites in the early life stages of marbled crayfish.

Comparative study of stress responses, laterality and familiarity recognition between albino and pigmented fish.

Oculocutaneous albinism is the result of a combination of homozygous recessive mutations that block the synthesis of the tyrosine and melatonin hormones. This disability is associated with physiological limitations, e.g., visual impairment expressed by lower visual acuity and movement perception, and eventually leads to acrophobia and/or photophobia, suggesting a potentially higher stress level associated with the behavioral responses of individuals with albinism to external stimuli compared to their pigmented conspecifics. However, in fish, differences in behavioral and/or physiological responses and stress levels between these phenotypes have been poorly documented. While acoustic perception of albino individuals is well known, the use of olfactory sensors for social communication, e.g., for the preference for familiar conspecifics, remains poorly understood. We performed two laboratory experiments with albino and pigmented European catfish Silurus glanis to observe: i) their behavioral and physiological responses to short-term stress induced by a combination of air exposure and novel environmental stressors and ii) their ability to use odor keys to recognize of familiar conspecifics and the influence of lateralization on this preference. In response to stress stimuli, albino fish showed higher movement activities and ventilatory frequencies and more often changed their swimming directions compared to their pigmented conspecifics. Blood plasma analysis showed significantly higher values of stress-, deprivation-, and emotional arousal-associated substances, e.g., glucose and lactate, as well as of substances released during intensive muscle activity of hyperventilation and tissue hypoxia, e.g., hemoglobin, mean corpuscular hemoglobin, erythrocytes, and neutrophil granulocytes. A preference test between environments with and without scented water showed the preference by both albino and pigmented catfish for environments with scent of familiar conspecifics, and both groups of fish displayed left-side lateralization associated with the observation of conspecifics and group coordination. The results tended to show higher physiological and behavioral responses of albinos to stress stimuli compared to the responses of their pigmented conspecifics, but the uses of olfactory sensors and lateralization were not differentiated between the two groups.

Acute toxicity of praziquantel to fish Danio rerio and planktonic crustacean Daphnia magna.

This study evaluated the toxicity of the pyrazino isoquinoline anthelmintic praziquantel (PZQ) to the Danio rerio zebrafish and Daphnia magna water flea. The estimated 24 h and 96 h LC50 of PZQ to the zebrafish was 39.9 mg/l and 30.4 mg/l, respectively. The highest 24 h and 96 h non-lethal concentration (LC0) was 21.7 mg/l and 21.2 mg/l, respectively. The mobility inhibition test of the juvenile Daphnia magna revealed a 48 h EC50 of 42.7 mg/l.

Effects of long-term exposure of Mytilus galloprovincialis to thiacloprid: A multibiomarker approach.

Thiacloprid is a neonicotinoid insecticide widely exploited in agriculture and easily mobilized towards aquatic environments by atmospheric agents. However, little information about its toxicological effects on aquatic invertebrate bioindicators is available. In this study, specimens of the mussel Mytilus galloprovincialis were exposed to thiacloprid at environmental (4.5 µg L-1) and 100 times higher than environmental (450 µg L-1) concentrations for 20 days. Thiacloprid affected haemolymph biochemical parameters, cell viability in the digestive gland, antioxidant biomarkers and lipid peroxidation in the digestive gland and gills at environmentally relevant concentrations (4.5 µg L-1). In addition, thiacloprid exposure caused histological damage to the digestive gland and gills. Interestingly, the pesticide was detected at levels equal to 0.14 ng g-1 in the soft tissues of sentinels exposed for 20 days to 450 µg L-1 thiacloprid in seawaterµ. Due to its harmful potential and cumulative effects after long-term exposure of M. galloprovincialis, thiacloprid may pose a potential risk to nontarget aquatic organisms, as well as to human health. This aspect requires further in-depth investigation.

Nitrogen factor of common carp Cyprinus carpio fillets with and without skin.

Consumer protection against food adulteration and misleading labelling is integrated into EU legislation, but accurate analysis of the meat content of farmed freshwater fish products is not possible because of the lack of established nitrogen factors for farmed common carp. The aim of this study was to determine nitrogen factors for farmed common carp Cyprinus carpio. Seven-hundred samples collected in 2018-2019 in three harvest seasons (March/April, Jun/July, and October/November) at seven locations in the Czech Republic were analysed for nitrogen, dry matter, protein, ash, and fat content according to standard ISO methods. The recommended nitrogen factor for fat-free common carp fillet with skin is 3.04 ± 0.13 and, for fillet without skin, 2.95 ± 0.12. Availability of nitrogen factors for common carp can help ensure that consumers are purchasing correctly labelled products.

Opening a new gateway towards the applications of chitosan nanoparticles stabilized Pickering emulsion in the realm of aquaculture.

In the current study, we synthesized eugenol (EU) based Pickering emulsion (PE) stabilized by food grade ingredients such as chitosan (CS) and tripolyphosphate (TPP) not only to enhance water miscibility of EU but also to decrease stress and damage to the immune system of fish due to anesthetic procedures. The formulated EUPEs were characterized in terms of droplet size, size distribution and the effects of environmental conditions e.g. pH and temperature on the behavior of the EUPEs. The results showed that EU PEs with 5% EU had smaller size with uniform distribution and were stable in the range of pH 5-7.5 and temperature 30-80 °C. The anesthetic effect of the EUPE was investigated by taking Common carp as a sample species. Interestingly, it was found that the induction time to anesthesia and recovery for the fish that received the PE was significantly shorter than that received EU at the same eugenol concentration (50 ppm). Most importantly, the improved hematological and bio-chemical parameters in the PE group further confirmed the immuno-protective and stress control efficacy of the PE. The results of this study propose a novel useful and potential application of PE in fishery where sedation is needed.

The effect of chronic exposure to chloridazon and its degradation product chloridazon-desphenyl on signal crayfish Pacifastacus leniusculus.

The effects of chloridazon (Ch) and its metabolite chloridazon-desphenyl (Ch-D) at the environmentally relevant concentrations of 0.45 µg/L and 2.7 µg/L on signal crayfish Pacifastacus leniusculus were assessed in a 30-day exposure followed by a 15-day depuration period. Locomotion, biochemical haemolymph profile, oxidative and antioxidant parameters, and histopathology were evaluated. Crayfish exposed to Ch at 0.45 µg/L and 2.7 µg/L showed significantly (p < 0.01) higher CAT activity and GSH level in hepatopancreas and gill compared to controls. The concentration of Ch at 2.7 µg/L was associated with significantly (p < 0.01) higher levels of GLU, LACT, ALT, AST in haemolymph compared to controls. Chloridazon-desphenyl exposure at both tested concentrations caused significantly higher (p < 0.01) GLU, LACT, ALT, AST, NH3, and Ca in haemolymph; lipid peroxidation (TBARS) levels in hepatopancreas; and CAT activity and GSH level in hepatopancreas and gill. Alterations of structure including focal dilatation of tubules, increased number of fibrillar cells, and haemocyte infiltration in the interstitium were observed with 2.7 µg/L Ch and with both Ch-D exposures. Locomotion patterns did not vary significantly among groups. A 15-day recovery period was insufficient to restore normal physiological parameters in exposed groups. Chloridazon and its metabolite Ch-D exerts harmful effects on crayfish.

The effects of the herbicides terbuthylazine and metazachlor at environmental concentration on the burrowing behaviour of red swamp crayfish.

Despite their low concentrations in many aquatic environments, evidence exists to suggest that herbicides do affect non-target organisms. Given that burrowing is a primary life-history trait in crayfish, herbicides could potentially have serious negative effects on these ecologically important freshwater macroinvertebrates. In this study, we exposed the red swamp crayfish Procambarus clarkii to terbuthylazine (a triazine) and metazachlor (a chloroacetanilide) at an environmental concentration of 2.0 µg/L for 28 days, and then observed their burrowing behaviour for two days. The metazachlor-exposed males excavated a greater number of burrows than the other tested groups, with comparable depths and volumes relative to individual specimen weight. The relative depth and volume of female burrows were identical in all groups. The natural habit of female crayfish of constructing deeper burrows than males was marginally significant in the control and META groups but was not significant for relative volume. The hypothesized adverse effects of chronic exposure to real environmental concentrations of herbicides were not documented in terms of either relative depth or volume. However, the increased number of burrows in metazachlor-exposed animals may mean that this invasive species will cause greater damage to embankments and river banks. The mechanisms behind these effects require closer study.

Pupil size variation as a response to stress in European catfish and its application for social stress detection in albino conspecifics.

Hormonal changes such as increased cortisol level in blood plasma in response to stress and social environmental stimuli are common among vertebrates including humans and typically accompanied by other physiological processes, such as changes in body pigmentation and/or pupil dilatation. The role of pupil size variation (PSV) as a response to stress have yet to be investigated in fish. We exposed albino and pigmented European catfish to short-term stress and measured changes in pupil size and cortisol level. Albinos showed lower pupil dilatation and higher cortisol levels than did pigmented conspecifics. A clear positive relationship between pupil dilatation and cortisol concentrations was observed for both pigmented and albino specimens, suggesting that PSV can be used as a stress indicator in fish, irrespective of albino's inability to express social communication by coloring. During the follow-up, we investigated whether a penultimate contest between albino individuals would impact contestants' social stress during subsequent contact. We observed PSV during the contact of unfamiliar albino catfish with different penultimate experiences (winner (W) and/or loser (L)). Then, the following treatment combinations were tested: WW, WL and LL. Twenty-four-hour contact of two unfamiliar catfish resulted in higher pupil dilatation among individuals with previous winner experience. Among treatment combinations, a WL contest displayed the highest pupil dilatation for winners. PSV reflected socially induced stress in individuals that was accompanied by the "winner" experience and dominancy in albinos. To conclude, the present study validates pupil dilatation as a non-invasive method to evaluate stress level in pigmented as well as albino fish in various contexts.