An integrative data-centric approach to derivation and characterization of an adverse outcome pathway network for cadmium-induced toxicity.

Cadmium is a prominent toxic heavy metal that contaminates both terrestrial and aquatic environments. Owing to its high biological half-life and low excretion rates, cadmium causes a variety of adverse biological outcomes. Adverse outcome pathway (AOP) networks were envisioned to systematically capture toxicological information to enable risk assessment and chemical regulation. Here, we leveraged AOP-Wiki and integrated heterogeneous data from four other exposome-relevant resources to build the first AOP network relevant for inorganic cadmium-induced toxicity. From AOP-Wiki, we filtered 309 high confidence AOPs, identified 312 key events (KEs) associated with inorganic cadmium from five exposome-relevant databases using a data-centric approach, and thereafter, curated 30 cadmium relevant AOPs (cadmium-AOPs). By constructing the undirected AOP network, we identified a large connected component of 18 cadmium-AOPs. Further, we analyzed the directed network of 59 KEs and 82 key event relationships (KERs) in the largest component using graph-theoretic approaches. Subsequently, we mined published literature using artificial intelligence-based tools to provide auxiliary evidence of cadmium association for all KEs in the largest component. Finally, we performed case studies to verify the rationality of cadmium-induced toxicity in humans and aquatic species. Overall, cadmium-AOP network constructed in this study will aid ongoing research in systems toxicology and chemical exposome.

In-feed emamectin benzoate abuse affects the biological responses and erythrocytes of Nile tilapia Oreochromis niloticus.

In aquaculture, drugs are often abused to accomplish disease control without considering the negative effects on fish health. This study aimed at elucidating the pernicious effects of in-feed antiparasitic drug emamectin benzoate (EB) abuse on the haemato-biochemistry and erythro-morphometry of healthy Nile tilapia Oreochromis niloticus. The fish were fed EB at 50 µg (1×) and 150 µg/kg biomass/d (3×) for 14 d as against the recommended 7 d and periodically assessed the blood parameters. A significant dose- and time-dependent reduction in feed intake, survival, total erythrocytes (TEC), monocytes (MC), hemoglobin (Hb), hematocrit (Ht) and mean corpuscular Hb concentration were noted. The total leukocytes (TLC), thrombocytes (TC), lymphocytes (LC) and neutrophils (NC) markedly augmented. The EB-dosing altered the fish physiology by enhancing the glucose, alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP) and creatinine and reducing the calcium, chloride and acetylcholinesterase (AChE) levels dose-dependently. The fish recovered within 4 weeks in the 1× group post-dosing but persevered in the overdosed group. The erythro-cellular and nuclear dimensions were reduced with the increase in dose and normalized after the cessation of dosing, except for nuclear volume. The erythro-morphological alterations were more prominent in the overdosed group. The results implied the pernicious effect of oral EB medication on the biological responses of fish if abused.

Impacts of Oral Florfenicol Medication and Residues on the Kidney and Liver of Nile Tilapia Oreochromis niloticus (L.)

Florfenicol (FFC), an approved aquaculture antibiotic, is administered in feed at doses of 10-15 mg kg biomass-1 day-1 for 10 successive days. In this study, healthy Oreochromis niloticus were fed with 0-10 times the therapeutic dose of 15 mg kg biomass-1 day-1 for 10 days and tracked for 43 days post dosing. Assessments of residue accrual and depletion, oxidative stress, serum biochemistry, histopathology and extent of kidney and liver damages were made. FFC dosing reduced the feed intake significantly. The therapeutic dose produced no mortalities on day 10. Dose-dependent alterations in serum biochemistry were noted upon dosing. Several histopathological alterations were observed in the kidney and liver, which vindicated the toxic potentials of FFC. The residual FFC and florfenicol amine (FFA) accrual, depletion and oxidative stress responses, such as increased malondialdehyde, total nitric oxide, ferric reducing antioxidant power and reduced glutathione S-transferase activity, were documented. The dietary FFC persuaded the physiological state of O. niloticus, the effects of which normalized sparsely with time upon cessation of dosing at the higher doses. The study provided a brief outlook on the physiological responses upon oral FFC administration, which should be kept in mind during its application for fish health safety purposes.

Histopathological changes and tissue residue concentrations of monosex Nile tilapia (Oreochromis niloticus, L) fries exposed to oxytetracycline.

The current study evaluated the biosafety of oxytetracycline (OTC) exposure for 30 days in monosex Oreochromis niloticus fries. The fries were exposed to OTC for 3 h/day for 30 days at 350 (0.5X), 700 (1X), 2100 (3X), 3500 (5X), and 7000 (10X) mg/L and compared with control (0X). The OTC exposure at 5X and 10X concentrations caused 100% mortality within 4 days and 5 min, respectively. The mortalities recorded in 0.5X, 1X, and 3X groups were 3.33 ± 1.15%, 14.67 ± 1.15%, and 47.33 ± 11.37% on day 30, respectively. The feed intake was decreased up to 23.33% in the 3X group during the exposure period. The OTC residue levels on 30-day exposure were 216.53 ± 14.71, 450.56 ± 44.31, and 1141.26 ± 63.64 µg/kg, which reduced to 40.40 ± 3.25, 76.68 ± 2.77, and 95.61 ± 5.13 µg/kg after 15 days of termination of exposure in the 0.5X, 1X, and 3X groups, respectively. The histopathological changes observed in the 1X group were epithelial detachment, desquamation of secondary lamellar epithelium, lamellar fusion, and inflamed cartilaginous core in the gills, alteration in the integrity of gut mucosa, degeneration of muscularis mucosae and necrosis in the intestine, the disintegration of the nephritic tubule, necrosis, and glomerulopathy in the kidney, and dilated vascular duct, necrotized hepatic tissue, diffused hepatic parenchyma, vacuolation, and fatty changes in the liver. The OTC exposure induced marked tissue changes histologically in a dose- and time-dependent manner, which undoubtedly reduced the growth of tilapia. Supplementary Information: The online version contains supplementary material available at 10.1007/s10499-022-00892-w.

The effects of extended feeding of florfenicol coated medicated diets on the safety, serum biomarkers and blood cells morphology of Nile tilapia Oreochromis niloticus (L.).

Tilapia is one of the most consumed farmed fish, which requires the use of antibiotics in certain phases of its production. This study assessed the safety of 30 days of oral florfenicol (FFC) dosing at 0-10 times the therapeutic dose (1 × : 10 mg/kg biomass/day) in Oreochromis niloticus juveniles. Behavioural changes, feed consumption, mortality and biomass were evaluated. Besides, the levels of serum glucose, calcium, chloride, creatinine, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase and blood cell morphology were determined at scheduled intervals. The 30 days of oral FFC dosing caused 3.33% (1 ×) to 18.33% (10 ×) mortalities, reduced feed intake and biomass in a dose-dependent manner. The fish fed the therapeutic dose recorded 1.25-fold increase in biomass, while the control group recorded 1.45-fold increase in 30 days. No significant erythrocyte morphological alterations were observed in the 1 × group compared to the control. However, marked morphological alterations like tear-shaped, spindle-shaped and degenerative erythrocytes in higher dosing groups indicated FFC cytotoxicity. All the serum biomarkers of O. niloticus increased significantly on day 10 and day 30 FFC dosing in a dose-dependent manner, except for calcium and chloride, which reduced significantly during the dosing period. Within 2 weeks of suspension of FFC dosing, the serum biomarker levels became normal except for alkaline phosphatase and creatinine. The recovery of biomass, feed intake, serum biomarker levels and erythrocyte morphological changes suggested that the FFC-induced changes are reversible. This study has, thus, proclaimed the safety of FFC at the therapeutic dose in O. niloticus.

Influence of dietary emamectin benzoate on the biological responses of monosex (all-male) Oreochromis niloticus (L.) fries.

The application of antiparasitic drugs plays a crucial role in the removal of infectious parasites in aquaculture. Emamectin benzoate (EB) is predominantly used as a feed premix against ectoparasites on temperate fish. This study evaluated the influence of 14 days of EB-dosing at 0-10 times the recommended dose (1X: 50 µg/kg biomass/day) on the biological responses and accrual/depletion of EB-residues in a tropical fish monosex Oreochromis niloticus fries. A significant dose-dependent reduction in feed intake by 3.50% in 1X and 43.00% in 10X groups, and an increase in mortalities from 2.92% (1X) to 11.25% (10X) during the EB-dosing period was noted. A significant increase in glucose and alkaline phosphatase and reduction in calcium and chloride ions, superoxide dismutase (SOD) and acetylcholinesterase levels in the muscle and/or brain tissue was observed. On day 21 post-EB-dosing, the levels of muscle glucose and SOD reached normalcy in the 1X group, while the levels of other biomarkers failed to recuperate. The EB-residue levels peaked on day 14 EB-dosing (2.77 ng/g) in the 1X group and decreased later with detectable levels (0.03 ng/g) even on day 21 post-EB-dosing. The EB-residue levels were within the permissible limits of the Canadian Food Inspection Agency and the European Commission. The EB-dosing negatively influenced the health of O. niloticus by altering the physiological state in a dose- and time-dependent way. The results suggested that the use of EB might be plausibly risky in tropical aquaculture.

Accumulation and clearance of tissue residues and health status of Nile tilapia Oreochromis niloticus (L.) juveniles as influenced by the extended oral oxytetracycline-dosing.

Antibiotics are considered an important primary therapy for bacterial diseases in aquaculture. This study evaluated the influence of oral administration of oxytetracycline (OTC) on feed intake, growth, mortality, residue accumulation and clearance, and histopathological changes in the vital organs of six groups of Nile tilapia Oreochromis niloticus when fed at 0-10 times the therapeutic dose (1×: 80 mg/kg biomass/day) for 10 and 20 consecutive days. The feed intake was reduced only slightly, viz., 2% in 10-day and 4.25% in 20-day dosing trials at 1× dose compared to control. While in other groups, an OTC-dose-dependent reduction in feed intake up to 31.25% was noted. The fish of the 0.5× and 1× groups recorded significantly high biomass, while the other OTC-dosed groups recorded significantly lower biomass than the control. The fold change in biomass between the control and 1× groups was insignificant. Dose-dependent mortalities were recorded in OTC-dosed fish in 10-day (1.67-6.67%) and 20-day (3.33-8.33%) trials. The OTC concentration in fish muscle established a dose- and time-response relationship. The OTC residue levels in muscle even on day 20 OTC-dosing were lower than the maximum residue limit (MRL) permitted by Codex Alimentarius (200 ng/g). On day 23 post OTC-dosing, the residue levels were traces to <10 µg/g in all groups, except the 10× group. The OTC-dosing caused mild to moderate pathological changes in the gills, liver and kidney of O. niloticus and the fish were able to mount adaptive biological responses to overcome the stress with time.

Dietary therapeutic dose of oxytetracycline negatively influences the antioxidant capacity and immune-related genes expression in Nile tilapia Oreochromis niloticus (L.).

Effects of the dietary therapeutic dose of oxytetracycline (OTC) at 80 mg/kg biomass/day for consecutive 10 days on the behaviour, feed intake, mortality, residue accumulation and depletion, antioxidant capacity and immune-related genes expression in juvenile Nile tilapia Oreochromis niloticus were evaluated. OTC-dosing caused mortalities, reduced feed intake, and biomass reduction at 24.5-28.5 °C. OTC residues recorded on day 10 (161.40 ± 11.10 ng/g) were within the maximum residue limits of the Codex Alimentarius. The withdrawal period was 7 days as per the European Commission's regulation. Traces of residues were present even on day 42 post-OTC-dosing. Dietary OTC reduced the antioxidant capacity of the liver and muscle tissues and down-regulated the expression of tumour necrosis factor-α, interleukin-1ß, and heat shock protein-70 genes in the liver significantly during the dosing period. The data generated on the biosafety of OTC-dosing may offer inputs for the development of management strategies in maintaining fish health and food safety.

Influence of Fluctuating Water Temperature and Dietary Oxytetracycline on the Safety of Monosex Nile Tilapia Oreochromis niloticus Fries.

The influence of fluctuating water temperature and dietary oxytetracycline (OTC) at 0 (0X), 80 (1X), 240 (3X), 400 (5X) and 800 mg (10X)/kg biomass/day for 30 consecutive days on the safety of monosex (all male) Nile tilapia Oreochromis niloticus fries in terms of feeding, growth, survival and histopathology of vital organs were assessed. A dose-dependent decline in feed intake and biomass was recorded. The OTC-dosed groups recorded higher mortalities than the control. The therapeutic OTC-dosing (1X) in conjunction with low temperature caused 75.56 ± 8.01% mortality and 25.75% reduced feed intake in 30 days. The mortalities increased with increasing OTC-doses from 85.19 ± 3.39% (1X) to 95.56 ± 2.22% (10X) and fluctuating temperature (12.00-21.50°C) even after the withdrawal of OTC. Relatively mild to moderate histopathological lesions were observed in the kidney, liver and intestine of OTC-dosed fries. These results suggested that dietary OTC and low water temperature may cause adverse effects on monosex O. niloticus fries.

Safety of emamectin benzoate administered in feed to Nile tilapia Oreochromis niloticus (L.).

Emamectin benzoate (EB) premix top-coated onto feed is extensively used to treat ectoparasitic crustacean infestations in aquaculture. This study evaluated the safety of EB-dosing in Nile tilapia Oreochromis niloticus at the recommended dose and dosage of 50 µg/kg biomass/day for 7 consecutive days (1X) and compared with control and 10 times the recommended dose (10X). Depletion of EB-residues in the edible muscle of 1X-dosed Nile tilapia was also studied. Mortality, behavioural changes, feed consumption, biomass, EB-residue depletion, and histopathological alterations in the kidney, liver and intestine were determined at slated intervals. Significant dose-dependent reduction in feed intake and biomass and insignificant mortalities were noted in 1X and 10X EB-dosed fish. In 1X EB-dosed fish muscle, the residues peaked on day 7 EB-dosing (9.72 ng/g) and decreased subsequently. Nevertheless, the residue levels were within the acceptable limit of the European Commission and the Canadian Food Inspection Agency even during the EB-dosing period. Histologically, tubule degeneration in the kidney, mild glycogen vacuolation in the liver, and loss of absorptive vacuoles, inflammation and disintegration of the epithelial layer in the intestine of Nile tilapia fed the 1X EB-diet were observed. The fish reverted back to their normal functions with time upon termination of oral-EB-dosing. This work contributed scientific data on the safety of EB particularly on the feed intake, growth reduction, mortality, histopathological alterations, and EB-residue levels in the edible tissues of Nile tilapia fed at the recommended dose and dosage, which suggested that EB-therapy might be reasonably risky in a tropical climate.

Dietary influences of oxytetracycline on the growth and serum biomarkers of Oreochromis niloticus (L.).

Tilapias are cultured globally and are rising in acceptance as the most important freshwater aquaculture species. Monitoring of serum biomarkers is a promising tool in aquaculture to screen the health status as they are virtuous indicators of extreme stress and organ dysfunction in fish. The present study examined the serum biomarkers of oxytetracycline (OTC)-dosed Nile tilapia Oreochromis niloticus at 0, 80 and 800 mg/kg biomass/day, i.e., 0X, 1X, and 10X the approved dose (X = 80 mg OTC/kg biomass/day) for 10 consecutive days. The fish biomass and levels of serum glucose, aspartate aminotransferase, alanine aminotransferase, creatinine and C-reactive protein (CRP) were determined at scheduled intervals. A significant dose-dependent reduction in fish biomass during the OTC-dosing (5.84%) and post-OTC dosing (8.16%) periods was observed. All the serum biomarkers of Nile tilapia increased significantly on day 10 OTC-dosing. Though their levels reduced significantly, normalcy was not achieved even after 42 days of cessation of OTC-dosing, except CRP. The CRP reached the normal level on day 25 post-OTC dosing in the 1X group. The results, thus, demonstrated that the oral OTC-dosing influences the physiological state of apparently healthy Nile tilapia in a dose-dependent manner. These changes were, however, reversible upon discontinuation of OTC-dosing. The set of data observed on growth reduction and elevated serum biomarker levels even after 42 days of cessation of OTC-dosing, thus, raises questions on the utility of oral OTC-dosing.