Toxicity Tests Research Articles

ESSAI DE TRAITEMENT BIOLOGIQUE DES HUILES LUBRIFIANTES USAGÉES RÉCUPÉRÉES DANS DES GARAGES DE KINSHASA À L’AIDE DES URINES HUMAINES ET KAOLIN EN RÉPUBLIQUE DÉMOCRATIQUE DU CONGO

In the various car garages of Kinshasa (in the Democratic Republic of Congo), the lack of consideration of environmental standards is the basis of the voluntary and/or accidental discharges of used oils directly into the environment without any prior treatment. These used hydrocarbons are pollutants that can cause major environmental problems due to their toxicity. The aim of this study is to treat used lubricating oils from car garages biologically in order to reduce their toxic potential. To this end, an aerobic biological treatment system was set up by mixing 4L of used oils with IL of urine, 2Kg of active Kaolin and 20L of water. The reduction in the toxicity of hydrocarbons was assessed through acute toxicity tests on the species Gambusia Affinis and microbiological analyses before and after biological treatment of these oils. The results obtained on the toxicity before treatment of these oils revealed that the LC50 was around ±3 ml/g of used oils to say that the pollutant is very toxic. After biological treatment, the LC50 rise to around ±66.7 ml/g for the treated oils, which represents a significant reduction in toxicity and ranks these treated oils in the order of less toxic effluents. The microbiological analyses of the used oils before their treatment did not show any bacteria; on the other hand, the analyses after treatment made it possible to highlight the presence of bacterial colonies whose identification revealed the following bacteria: Citrobacter, Enterobacter, Klebsiella and Pseudomonas, which are mostly lipolytic. This study thus allowed to treat used oils thanks to microorganisms and the intrinsic properties of urine and Kaolin.

Safety Assessment and Evaluation of Probiotic Potential of Lactobacillus bulgaricus IDCC 3601 for Human Use

Lactic acid bacteria (LAB) are probiotic microorganisms widely used for their health benefits in the food industry. However, recent concerns regarding their safety have highlighted the need for comprehensive safety assessments. In this study, we aimed to evaluate the safety of L. bulgaricus IDCC 3601, isolated from homemade plain yogurt, via genomic, phenotypic, and toxicity-based analyses. L. bulgaricus IDCC 3601 possessed a single circular chromosome of 1,865,001 bp, with a GC content of 49.72%, and 1910 predicted coding sequences. No virulence or antibiotic resistance genes were detected. Although L. bulgaricus IDCC 3601 exhibited antibiotic resistance to gentamicin and kanamycin, this resistance is an intrinsic feature of this species. L. bulgaricus IDCC 3601 did not produce biogenic amines and did not exhibit hemolytic activity. Phenotypic analysis of enzyme activity and carbohydrate fermentation profiles revealed the metabolic features of L. bulgaricus IDCC 3601. Moreover, no deaths or abnormalities were observed in single-dose oral toxicity tests, suggesting that L. bulgaricus IDCC 3601 has no adverse effect on human health. Finally, L. bulgaricus IDCC 3601 inhibited the growth of potential carbapenem-resistant Enterobacteriaceae. Therefore, our results suggest that L. bulgaricus IDCC 3601 is a safe probiotic strain for human consumption.

Phytochemical Screening and Safety Assessment of a Polyherbal Tea Formulation of Lippia Multiflora, Zinger Officinale and Mentha Piperita

Aims: Lippia multiflora, Zingiber officinale, and Mentha piperita are aromatic plants known for their health benefits. This study the phytochemical composition and evaluate the safety of a tea made from them for safe human consumption. Methodology: For tea formulations, powder of each plant was prepared and weighed in specific proportions to formulate five different polyherbal tea. A hedonic test was then conducted on each formulation to evaluate consumer apprecation. Phytochemical composition was carried out by coloration and precipitation tests. This was followed by acute and subacute toxicity tests. Results: Among the five formulations, formulation 4 consisting of “55% L. multiflora, 30% Z. officinale and 15% M. piperita" was the most appreciated tea with a value of 8.01. Regarding safety of the formulation 4, no deaths or signs of intoxication were observed. Subacute toxicity tests showed that the polyherbal tea 4 has no adverse effects on biochemical parameters. Apart from white blood cells for which there was an increasing number from 14.40 ± 0.50 103/uL to 19.30 ± 0.40 103/uL in comprarison to the control group (10.60 ± 0.90 103/uL), the haematological parameters were not significantly modified. This Tea formulation ²was rich in compounds such as alkaloids, polyphenols, flavonoids, saponosides, sterols and polyterpenes, leucoanthocyanins, and mucilages, which made its consumption more attractive. Conclusion: The polyherbal tea 4 which was the most appreciated, was rich in secondary metabolites such as alkaloids, polyphenols, flavonoids, saponosides, sterols and polyterpenes, leucoanthocyanins, and mucilages, and was not toxic to wistar rats.

Advancements in ecotoxicological assessment of anticancer agents: Bridging sustainability and innovation in chemical testing

Ecotoxicological testing is the latest buzzword in the field of environmental testing and monitoring the effect of pharmaceuticals on the ecosystem. Testing alternative and innovative study models enriches the ecotoxicological perspective by introducing greenness and sustainability into the ecosystem. Pharmaceuticals, being a major source of ecosystem contamination, necessitate the practice of such approaches. The current review systematically discusses the advancements and paradigm shift from traditional to greener ecotoxicity testing of important classes of pharmaceuticals, such as anticancer agents. Additionally, it discusses the approaches for developing sustainable pharmaceuticals from anticancer agents. The common approaches and future possibilities further demonstrate the importance of this holistic approach, which can efficiently bridge the gap between ecosystem sustainability and toxicity testing related to them.

Galleria mellonella as an Antimicrobial Screening Model.

To combat the rising global issue of antibiotic resistance, the accelerated development of novel antibiotics is essential. Current preclinical antimicrobial development yields a significant number of leads that prove unsuitable either prior to or during clinical trials. To increase the efficiency of preclinical development, relevant, standardized, accessible, and cost-effective models must be developed. Galleria mellonella (greater wax moth) larvae are widely used as an infection model to assess microbial virulence, conduct drug toxicity testing, and serve as a preliminary means of evaluating the in vivo efficacy of novel antimicrobial compounds. These infection models have greater biological relevance than many in vitro screens of comparable throughput and decrease reliance on mammalian models when used as a pre-screen for antimicrobial testing. This protocol describes a standardized methodology for the optimization of G. mellonella infection models, which can be applied to bacterial species and antimicrobial therapeutics of choice. Using the WHO priority pathogen Pseudomonas aeruginosa as an exemplar, we outline steps that can be undertaken to develop a reproducible model of infection and therapeutic testing. This includes recommendations on experimental setup, sample preparation, and infection and treatment protocols. Integration of this model within preclinical antimicrobial development pipelines would decrease reliance on mammalian models, reduce the number of ineffective compounds reaching clinical trials, and ultimately increase the efficiency of preclinical antimicrobial development.

Literature Review: Guava Leaves (Psidium Guajava) for Treating Diarrhea

Diarrhea is a significant health problem, especially in developing countries. Guava leaves (Psidium guajava) have long been used in traditional medicine to treat diarrhea. This study aims to review the literature on the effectiveness of guava leaves in treating diarrhea by focusing on ten aspects: raw material name, raw material content, raw material composition, active substance content, efficacy claims, empirical data, raw material standardization, preclinical test, toxicity test, and clinical trial. The results of several studies show that guava leaves contain tannins, flavonoids, saponins, and essential oils that play a role in the antidiarrheal effect. Preclinical research shows a reduction in the frequency of bowel movements and an increase in stool consistency in animal models. Although empirical claims and preclinical trials support the use of guava leaves as a treatment for diarrhea, more research is still needed regarding the standardization of raw materials, toxicity tests, and clinical trials to ensure its overall efficacy and safety.

Ecotoxicological risk assessment of active pharmaceutical ingredients (APIs) against different aquatic species leveraging intelligent consensus prediction and i-QSTTR modeling

The increasing presence of active pharmaceutical ingredients (APIs) in aquatic ecosystems, driven by widespread human use, poses significant risks, including acute and chronic toxicity to aquatic species. However, the scarcity of experimental toxicity data on APIs and related compounds due to the high costs, time requirements, and ethical concerns associated with animal testing hinders comprehensive risk assessment. In response, we developed quantitative structure-toxicity relationship (QSTR) and interspecies quantitative structure toxicity-toxicity relationship (i-QSTTR) models for three key aquatic species: zebrafish, water fleas, and green algae, using NOEC as an endpoint, following OECD guidelines. Algae, daphnia, and fish, recognized as standard organisms in toxicity testing, are crucial bio-indicators due to their size, transparency, adaptability, and regulatory acceptance. We used partial least squares (PLS) and multiple linear regression (MLR) methods for model development alongside machine learning techniques such as Random Forest (RF), Support Vector Machines (SVM), K-nearest Neighbor (kNN), and Neural Networks (NN) to enhance the predictivity. Lipophilicity, electronegativity, unsaturation, a molecular cyclized degree in molecular structure, large fragments, aliphatic secondary C(sp2), and R–CR–R groups were identified as critical biomarkers for API toxicity. Screening of the PPDB (pesticide properties databases) and DrugBank validated the practical application of these models, offering valuable tools for regulatory decisions, safer API design, and the preservation of aquatic biodiversity.

Development of Indonesian Original Medicines Study of Standardized Herbal Medicine Products "Antangin JRG" as Traditional Medicine

This study aims to examine the potential for the development of indigenous Indonesian medicines based on traditional medicinal plants. Herbal medicine has been used for generations in Indonesia and has great potential to be developed into more standardized and modern products. This study covers various aspects ranging from product name, content, class, composition, efficacy claims, empirical data, standardization of raw materials, preclinical tests, toxicity tests, to clinical trials. The results of the study show that with proper processing and standardization, Indonesian traditional medicine can compete with modern pharmaceutical products.

Genotoxicity Of The Herbicide Picloram (TORDON ® 24K) on tadpoles of Boana pulchella (Amphibia, Anura, Hylidae)

Picloram (PIC) is an herbicide widely used in Uruguay to control weeds, especially in forest crops. In the present study, we evaluate the acute toxicity and genotoxicity of formulated 24.9% PIC-based Tordon®24K on Boana pulchella tadpoles under standard laboratory conditions. In the acute toxicity test we use mortality as the endpoint and we obtained a 48-h LC50 of 16.5 mg/L, LOEC 14.5 mg/L, and NOEC 12.5 mg/L. We determined genotoxicity in GS36 tadpoles exposed to sublethal concentrations of 8.5 (NOEC/1.5) and 12.5 mg/L (NOEC) PIC using micronucleus frequency (MN) and occurrence of nuclear abnormalities (AN) as endpoints. We noticed a significant increase in the frequency of MN and AN in all treatments tested at sublethal concentrations. Our findings demonstrate the genotoxic effects of this herbicide, which represent a danger to amphibians that inhabit agroecosystems. Based on the results obtained and considering that Boana pulchella is not a threatened species and has a wide distribution in the Uruguayan territory and the Pampa Biome, it could be considered a reference organism in environmental monitoring programs in the region.

A tiered toxicity testing strategy for assessing early life stage toxicity in estuarine fish (Mugilogobius chulae): A case study on tris (1-chloro-2-propyl) phosphate ester

The estuarine ecological environment faces significant threats from contaminants of emerging concern (CECs); yet, the risk posed by CECs to resident organisms remains poorly understood. Here, we employed tiered toxicity testing to investigate the adverse effects and potential mechanisms of tris (1-chloro-2-propyl) phosphate (TCPP) on the early life stages of an estuarine fish, Mugilogobius chulae. TCPP affected the development of M. chulae embryos, including survival, morphology, hatching, and behavior. A concentration-dependent transcriptomic analysis showed that TCPP disrupted 12 neurodevelopment-related KEGG pathways in M. chulae embryos, with five of the 30 % top-ranked pathways related to neurotransmitter signaling. Besides the cholinergic synapse signaling pathway, the glutamatergic signaling pathway (including NMDAR and AMPAR subtypes) may also mediate TCPP-induced neurodevelopmental toxicity. The NMDAR subtype GRIN2B was downregulated at high concentrations. Molecular dynamics simulations revealed a strong interaction between TCPP and GRIN2B, with TCPP binding to the residues Ile153 and Ile188. The results suggest that NMDARs play a crucial role in TCPP-induced neurodevelopmental toxicity toward M. chulae. AOP network analysis predicted that TCPP may impact cognitive functions and memory. Our study provides a novel testing strategy for identifying the mechanisms of toxicity of CECs, a crucial component of ecological risk assessment.

Toxicodynamics of cadmium in the green mussel Perna viridis (Linnaeus, 1758) using bioenergetic and physiological biomarkers.

This study evaluated the toxicity of cadmium (Cd) on the green mussel Perna viridis, aiming to identify toxicological endpoints and investigate its responses across physiological, bioenergetic, and biochemical parameters. The 96-hour LC50 value for Cd in P. viridis was 3.03 ± 0.12 mg L-1, with a 95% confidence interval of 2.35-3.91 mg L-1. Chronic toxicity tests revealed a No Observable Effect Concentration (NOEC), Lowest Observable Effect Concentration (LOEC), and chronic toxicity values of 0.20, 0.37, and 0.29 mg L-1, respectively. Cadmium accumulation in treated mussels increased 46- to 215-fold compared to the control group. Superoxide dismutase, catalase, glutathione S-transferase, and glutathione peroxidase levels in exposed mussels exhibited a significant increase compared to the control group. The redox index ratio, acetylcholinesterase activity, and lysosomal membrane stability decreased with increasing exposure concentrations. Levels of reduced and oxidized glutathione, glutathione reductase, lipid peroxidation, and metallothionein-like proteins increased in exposed mussels. Clearance rate, respiration rate, and excretion rate decreased in a dose-dependent manner. Protein, carbohydrate, and lipid levels decreased with increasing exposure concentration (p < 0.001). Mitochondrial electron transport system activity increased, while cellular energy allocation (p < 0.001) and scope for growth decreased in a dose-dependent manner (p < 0.01). The significant increase in antioxidants suggests heightened oxidative stress in mussels under Cd exposure. The physiological activities of the mussels were severely affected, ultimately leading to a reduced scope for growth. The toxicological data generated in this study contribute to the development of seawater quality criteria for the metal Cd.

Biocompatibility evaluation and imaging application of a new fluorescent chemodosimeter for the specific detection of mercury ions in environmental and biological samples

Mercury is widely distributed in the natural environment and living environment, and as a heavy metal element with significant toxicity mercury ions pose a threat to agriculture and the food chain, thus consequently the residue and bioaccumulation of mercury ions will cause numerous and serious diseases, such as neurological abnormalities, gingivitis and tremor. Therefore, the development of a responsive and accurate fluorescent probe or sensor with excellent biocompatibility for mercury ions is crucial and marketable in the health and environmental protection. In this study, a novel fluorescent probe named BE-Hg has been innovatively proposed for the dynamic monitor of mercury ions. The probe showed excellent spectral performance in vitro experiments with detection limit as low as 1.18 nM, while it was applied to monitor environmental water samples with recovery as high as 93 %. Moreover, our group evaluated the probe safety using fluorescence-labeled tissue, organ and organ system including liver, heart, blood vessel and immune system in zebrafish for the first time. Note that the probe is proved with good biocompatibility by the cytotoxicity test, behavioral test, hepatotoxicity test, cardiotoxicity test, blood vessel toxicity test and immunotoxicity test with 15 indicators. In addition, the probe owns the ability to detect Hg2+ in live cells and zebrafish models. Thus it is expected to be a tool for the detection of mercury ions in water environments and organisms. In addition, behavioral test, hepatotoxicity test, cardiotoxicity test, blood vessel toxicity test and immunotoxicity test can be used to evaluate the biocompatibility of fluorescent probe, which provide a reliable safeguard for the research and development of probe.

The hanging‐heart chip: A portable microfluidic device for high‐throughput generation of contractile embryonic stem cell‐derived cardiac spheroids

AbstractStem cell‐derived cardiac spheroids are promising models for cardiac research and drug testing. However, generating contracting cardiac spheroids remains challenging because of the laborious experimental procedure. Here, we present a microfluidic hanging‐heart chip (HH‐chip) that uses a microchannel and flow‐driven system to facilitate cell loading and culture medium replacement operations to reduce the laborious manual handling involved in the generation of a large quantity of cardiac spheroids. The effectiveness of the HH‐chip was demonstrated by simultaneously forming 50 mouse embryonic stem cell‐derived embryonic bodies, which sequentially differentiated into 90% beating cardiac spheroids within 15 days of culture on the chip. A comparison of our HH‐chip method with traditional hanging‐drop and low‐attachment plate methods revealed that the HH‐chip could generate higher contracting proportions of cardiac spheroids with higher expression of cardiac markers. Additionally, we verified that the contraction frequencies of the cardiac spheroids generated from the HH‐chip were sensitive to cardiotoxic drugs. Overall, our results suggest that the microfluidic hanging drop chip‐based approach is a high‐throughput and highly efficient method for generating contracting mouse embryonic stem cell‐derived cardiac spheroids for cardiac toxicity and drug testing applications.

Ocular effects of synthetic cannabinoids: a case-control study.

To evaluate the ocular effects of "Bonzai", a synthetic cannabinoid (SC), in seropositive and seronegative users. Sixty eyes of 60 consecutive male patients with a history of "Bonzai" use and 30 eyes of 30 age-matched male healthy controls were enrolled in this case-control study. Patients with past "Bonzai" use were grouped as seropositive (n:30) and seronegative (n:30) according to urine toxicology tests. All groups were compared for blood pressures, intraocular pressure, foveal and parafoveal retinal thicknesses, subfoveal and parafoveal choroidal thicknesses, measurements of the peripapillary retinal nerve fibre layer (RNFL), and macular ganglion cell complex (GCC), subfoveal total choroidal, luminal and stromal areas, and choroidal vascularity index (CVI). No differences were noted in blood pressures between the groups (p > 0.05). The mean intraocular pressure was significantly lower in the seropositive group than in the other groups (p < 0.001). Foveal and retinal thicknesses, RNFL, and GCC measurements did not differ between the groups (p > 0.05). Subfoveal and parafoveal choroidal thicknesses and areas were lower in the seropositive group than in the other groups (p < 0.001, for all). CVI increased in both groups with "Bonzai" use compared to the control group (p < 0.001, for all). This study indicates that intraocular pressure may decrease, and choroidal changes may be observed in SC users. Further clinical studies with a larger sample size, especially using purified SC for therapeutic purposes, are needed to confirm the present findings, and further histopathologic studies are required to clarify the changes in the choroid despite SC seronegativity. NCT06235346.

EFFECTS OF USING HYDROXIAPATITE-GELATIN GEL AND ROSELLA EXTRACT TO PREVENT RELAPSE IN ORTHODONTIC TREATMENT

To correct malocclusion and malposition in dentistry using orthodontic treatment. Teeth that have moved through the bone with orthodontic devices tend to return to their original position or what is usually called relapse. Relapse is when the teeth return to their original position before orthodontic treatment was carried out. Relapse prevention can be achieved using mechanical devices but there are still some limitations. The use of scaffolds can be a solution in preventing tooth relapse because scaffolds can support cells to grow and develop into new bone in the damaged area. The scaffold components are hydroxyapatite, gelatin, and rosella flower extract which contains flavonoids to inhibit bone resorption. To find out whether these components are toxic or not before they are applied to the human body, a cytotoxicity test is carried out, which is a laboratory study. This research was carried out by incubating the hydroxyapatite scaffold, gelatin and rosella flower extract with osteoblast cells for 24 hours to see whether the rosella extract was toxic to osteoblast cells. Conclusion: This toxicity test shows that rosella flower extract as an ingredient in making scaffolds is not toxic to osteoblast cells.

Lab scale evaluation of biomineralization-assisted decolourization of reactive blue 4 and toxicity analysis

The process of microbial mineral precipitation has proven to be highly effective in addressing a wide range of pollutants in different environments. The objective of this study is to utilize the mineralization abilities of microbial strains found in the Noyyal River basin, which is home to a significant textile center in India, in order to both remove the color from reactive blue 4 and promote biomineralization. Promising results were obtained from the microbial isolates Streptomyces sp., Rhodococcus ruber, and Bacillus sp., and their consortium which demonstrated the ability to induce biomineralization and decolorize reactive blue 4. Characterization of the obtained biominerals through X-ray diffraction and FT-IR analysis showed the presence of well-crystallized polymorphs of calcium carbonate, along with other organic compounds. SEM-EDX analysis of biominerals formed during biomineralization-assisted decolourization of RB4 revealed distinct morphologies and elemental composition of the biominerals produced by microbial interactions with dye molecules. Further, to assess the safety and efficacy of these biominerals, zebrafish embryo toxicity tests were conducted by comparing biominerals produced during dye decolourization with the pure reactive blue 4 dye. Zebrafish embryos treated with biominerals exhibited normal developmental patterns, whereas those exposed to reactive blue 4 displayed mortality and abnormal hatching rates. The heart rates of the Zebrafish embryos were normal for controls and biomineral groups, but lower for RB4-exposed embryos. In conclusion, this study underscores the potential of biomineralization in remediating dye-contaminated wastewater, offering circular solutions for waste reduction and resource utilization.

In vivo immunomodulatory effect and safety of MSC-derived secretome

Background: Secretomes have been gaining interest in treating several diseases due to their pharmaceutical effects, such as the immunomodulatory effect. This study aimed to determine the immunomodulatory effect of secretomes derived from human umbilical cord mesenchymal stem cells (MSCs) and their safety. Methods: We conducted an in vivo immunomodulatory study using a carbon clearance assay. The safety of single-dose administration of secretome was done using fixed-dose methods of acute toxicity test. Results: The phagocytic index was higher in mice treated with secretome than in untreated mice. The acute toxicity study also showed that the administration of secretome derived from human umbilical cord MSCs did not change the mice’s body weight, physical examination results, organ weight, and gross anatomy examination. Conclusions: This study presents the potential of secretome derived from MSCs as a safe immunomodulatory agent.

In silico occupational exposure banding framework for data poor compounds in biotechnology.

Occupational exposure limits (OELs) and occupational exposure bands (OEBs) provide quantitative benchmarks for worker health protection. If empirical toxicology data are insufficient to derive an OEL, an OEB is often assigned using partial toxicology data along with other relevant hazard information. There is no consensus methodology to assign OEBs for chemicals lacking any empirical toxicology data. Thus, this study developed an in silico framework for OEB assignment of data poor compounds. It relies upon computational tools to evaluate standard toxicological end points and to assign reliability ratings, which are then used to assign Global Harmonization System (GHS) hazard categories. Subsequently, the hazard categories are entered into the National Institute for Occupational Safety and Health (NIOSH) occupational exposure banding tool to assign bands for individual end points as well as an overall OEB. As a proof-of-concept, five compounds with established OELs (i.e., "knowns") were evaluated. The knowns were assigned to overall OEBs C, D, or E, four of which were equal to or lower than the OEBs based on actual harmonized GHS categories as well as established OELs, indicating that the OEBs assigned using this framework are likely to be protective. Subsequently, five compounds with little to no experimental data and no established OELs from any U.S. agency or consensus OEL-setting organizations were evaluated (i.e., "unknowns"). The unknowns were assigned to overall OEBs D or E. It was concluded that the proposed framework can be used to assign protective OEBs to compounds with little to no toxicology testing data. As additional data become available, the compound may be de-risked, and a precautionary OEB (or an OEL) can be assigned. The proposed framework provides an example of a practical methodology to evaluate data poor compounds and shows that the output of this framework is expected to be protective of worker health.

Exposure to Cyantraniliprole Adversely Impacts Fitness of Harmonia axyridis: Acute Toxicity and Sublethal Effects on Development, Fecundity and Antioxidant Responses.

Extensive utilization of pesticides and their persistent residues inadvertently pose threats to the effectiveness and fitness of biocontrol agents in agroecosystems. However, these ecological consequences are generally disregarded when executing integrated pest management strategies (IPM). Cyantraniliprole (CNAP) serves as a wide-spectrum diamide insecticide and its sublethal effects have been well characterized on multiple insect pests, whereas its impacts on beneficial natural enemies remain unfathomed. Herein we exposed Harmonia axyridis, a predacious generalist, to lethal and sublethal concentrations of CNAP via dipping treatment (egg stage) and topical applications (1st-instar stage + adult stage). The acute toxicity tests revealed that LC50 of CNAP were 90.11, 86.11 and 240.50 mg/L against embryos, 1st instar nymphs and female adults, respectively, with safety factors ranging from 1.14 to 5.34, suggesting its medium toxicity for H. axyridis and larval stage was the most susceptible. The embryonic, larval and pupal durations of coccinellids ecdysed from CNAP-treated eggs and 1st instars were all elongated under sublethal concentrations, of which LC30 triggered more pronounced and significant retardations relative to control. Besides, exposed coccinellids displayed substantially diminished pupal mass and pupation rate, most notably for insects molted from the 1st-instar stage upon CNAP sublethal treatments. With respect to reproductive performance, LC10 and LC30 of CNAP all significantly suppressed female fecundity, as evidenced by reduced vitellin content, a prolonged pre-oviposition period (POP), mitigated laid eggs and the egg hatching rate. Specifically, there existed positive correlations between vitellin level (Vn) and number of eggs deposited by per female, indicative of CNAP affecting fecundity by regulation of Vn. In addition, the antioxidant system was also profoundly disrupted by CNAP, with compromised POD activity at different concentrations over time and induced hormesis of SOD/CAT activities post LC10 exposure. Activities of SOD and TAC were enhanced to exert protective functions during the first 48 h, while defense collapsed at 72 h following LC30 treatments that depleted all enzymatic activities. We speculated that fitness trade-offs may occur between reproductive capacity and antioxidant defenses to sustain physiological homeostasis in response to CNAP stress. Collectively, this study evaluated the ecological risk of CNAP and unmasked its adverse implications for overall fitness of H. axyridis, which highlighted rational application of agrochemicals to conserve biocontrol agents when implementing IPM strategies for sustainable pest control.

A Comparison of European Union and Turkish Reports on the Statistics of Experimental Animal Use in Toxicity Testing.

The data on the number of animals used for experimental purposes, with a particular focus on regulatory toxicity studies, in Turkey and EU member states plus Norway (EU+), were compared for the period 2015 to 2020 (UK data after Brexit for 2020 were included too). The total number of listed vertebrates used for regulatory toxicity testing in the full 6-year reporting period in Turkey was 3.6% of that reported for the EU+. However, these numbers showed an increasing trend over the reporting period in Turkey, while the trend decreased in the EU+. In the top three most-used species for regulatory toxicology purposes in Turkey, 41% were fish (68,758 animals), followed by rats (29%; 49,975 animals), and then mice (20%; 33,292 animals). Turkey used low numbers of cats and dogs, while the corresponding number for the EU+ was more than 70-fold higher. Non-human primates were not used at all in Turkey, whereas about 34,000 were used in EU+ laboratories. The majority of toxicity tests (57%) performed in Turkey were categorised as 'Other' toxicity tests, and 27% were acute and sub-acute toxicity tests. Successful replacement alternatives (e.g. in vitro and in silico approaches) are already used in a wide range of research areas. However, although this new research and testing paradigm is underpinned by scientific evidence, the legislation and pace of acceptance of these new techniques in Turkey is considerably lagging behind other countries.