Residual determination and acute toxicity of the neonicotinoid clothianidin in the neotropical stingless bee Tetragonisca angustula Latreille, 1811 (Apidae: Meliponini).

Bees play a crucial role as natural pollinators, ensuring the maintenance and stability of the world's biodiversity and agricultural crops. Native bees in neotropical regions belong to the Meliponini tribe, a larger group that differs significantly in behavior and biology from honeybees (e.g., Apis mellifera) and solitary bees (e.g., Osmia spp.). Hence, the exposure and effects of pesticides is also likely to vary among these different species. The aim of this study was to develop an analytical method to determine the presence of the neonicotinoid clothianidin in the Brazilian native stingless bee Tetragonisca angustula (local common name: Jataí). The method used for the chemical analysis involved a QuEChERS technique combined with UHPLC-MS/MS analysis. The developed method was subsequently used to analyze collected field samples. In addition, the acute toxicity of the pesticide to T. angustula was evaluated in a laboratory bioassay evaluating both lethal and sublethal endpoints. The analytical method was successfully developed with detection and quantification limits of 1.55 and 5 µg L-1, respectively, along with a linear range of 1-5 ng mL-1. Clothianidin was detected in environmental samples (9.2-32.9 ng g-1), and the exposure experiments demonstrated acute oral toxicity to adults of T. angustula, (24 h-LD50 of 0.16 ng a.i./bee), as well as no significative interference in acetylcholinesterase activity. Considering the obtained toxicity endpoints for T. angustula and those reported in the literature for other bee species, this study revealed that T. angustula is more (lethally) sensitive to clothianidin than other bee species, including those commonly used in environmental risk assessment studies. This thus also supports the call for using native test species in (regional) risk assessment evaluations.

Developmental cardiotoxicity of 4,5-dichloro-2-n-octyl-4-isothiazolin-3-one (DCOIT) in marine medaka (Oryzias melastigma).

The application of 4,5-dichloro-2-n-octyl-4-isothiazolin-3-one (DCOIT) as an antifouling biocide causes high toxicity to non-target marine organisms. To examine the developmental cardiotoxicity and mechanisms of DCOIT, we concurrently performed sub-chronic exposure and life-cycle exposure experiments using marine medaka embryos. After sub-chronic exposure to DCOIT at 1, 3, 10, and 33 µg/L, cardiac defects were caused by upregulation of cardiac gene transcriptions, decreasing heart size, and accelerating heartbeat. Hyperthyroidism in medaka larvae was identified as the cause of developmental cardiotoxicity of DCOIT sub-chronic exposure. In addition, parental life-cycle exposure to 1, 3, and 10 µg/L DCOIT led to transgenerational impairment of cardiogenesis in offspring medaka. A crossbreeding strategy discriminated a concentration-dependent mechanism of transgenerational cardiotoxicity. At 1 µg/L, the DCOIT-exposed female parent transferred a significantly higher amount of triiodothyronine (T3) hormone to offspring, corresponding to an accelerated heart rate. However, DCOIT at higher exposure concentrations modified the methylome imprinting in larval offspring, which was associated with cardiac dysfunction. Overall, the findings provide novel insights into the developmental cardiotoxicity of DCOIT. The high risks of DCOIT-even at environmentally realistic concentrations-raise concerns about its applicability as an antifoulant in a marine environment.

Isothiazolinone dysregulates the pattern of miRNA secretion: Endocrine implications for neurogenesis.

Isothiazolinones are extensively used as preservatives and disinfectants in personal care products and household items. The unintended exposure of humans and animals to isothiazolinones has led to increasing concerns about their health hazards. The compound 4,5-Dichloro-2-n-octyl-4-isothiazolin-3-one (DCOIT), a representative isothiazolinone, can simultaneously induce endocrine disruption and neurotoxicity. However, the underlying mechanisms and linkages remain unclear. Our purpose was to elucidate the role of miRNAs as the signaling communicator during the crosstalk between endocrine and nervous systems in response to DCOIT stress. H295R cells were exposed to DCOIT, after which the alterations in intracellular miRNA composition, exosome secretory machinery, and extracellular miRNA composition were examined. Then, a PC12 cell line of neuronal differentiation potential was cultured with the extract of extracellular miRNAs from DCOIT-exposed H295R cell media to explore the functional implications in neurogenesis. The results showed that DCOIT exposure resulted in 349 differentially expressed miRNAs (DEMs) in H295R cells, which were closely related to the regulation of multiple endocrine pathways. In the media of H295R cells exposed to DCOIT, 66 DEMs were identified, showing distinct compositions compared to intracellular DEMs with only 2 common DEMs (e.g., novel-m0541-5p of inverse changes in the cell and medium). Functional annotation showed that extracellular DEMs were not only associated with sex endocrine synchronization, but were also implicated in nervous system development, morphogenesis, and tumor. Incubating PC12 cells with the extracellular exosomes (containing miRNAs) from DCOIT-exposed H295R cells significantly increased the neurite growth, promoted neuronal differentiation, and shaped the transcriptomic fingerprint, implying that miRNAs may communicate transduction of toxic information of DCOIT in endocrine system to neurons. Overall, the present findings provide novel insight into the endocrine disrupting and neural toxicity of DCOIT. The miRNAs have the potential to serve as the epigenetic mechanism of systems toxicology.

Short communication: Is there a difference in the sensitivity of Africanised and European honeybees to thiamethoxam or dimethoate?

The similarity of sensitivity of adult Africanised and European honeybees following acute oral exposure to thiamethoxam has been questioned. Data collated from adult acute contact and oral toxicity testing of a range of thiamethoxam containing products (solo and mixtures) shows that the toxicity of these products to Africanised honeybees can be directly predicted from the toxicity of the active substances to European honeybees. Similarly, the acute contact and oral toxicity of dimethoate to Africanised bees lies within the same range as European honeybees. There are no major differences in the sensitivity of Africanised and European honeybee individuals to thiamethoxam and dimethoate.

Comparative toxicity of two neonicotinoid insecticides at environmentally relevant concentrations to telecoprid dung beetles.

Dung beetles (Coleoptera: Scarabaeinae) frequently traverse agricultural matrices in search of ephemeral dung resources and spend extended periods of time burrowing in soil. Neonicotinoids are among the most heavily applied and widely detected insecticides used in conventional agriculture with formulated products designed for row crop and livestock pest suppression. Here, we determined the comparative toxicity of two neonicotinoids (imidacloprid and thiamethoxam) on dung beetles, Canthon spp., under two exposure profiles: direct topical application (acute) and sustained contact with treated-soil (chronic). Imidacloprid was significantly more toxic than thiamethoxam under each exposure scenario. Topical application LD50 values (95% CI) for imidacloprid and thiamethoxam were 19.1 (14.5-25.3) and 378.9 (200.3-716.5) ng/beetle, respectively. After the 10-day soil exposure, the measured percent mortality in the 3 and 9 µg/kg nominal imidacloprid treatments was 35 ± 7% and 39 ± 6%, respectively. Observed mortality in the 9 µg/kg imidacloprid treatment was significantly greater than the control (p = 0.04); however, the 3 µg/kg imidacloprid dose response may be biologically relevant (p = 0.07). Thiamethoxam treatments had similar mortality as the controls (p > 0.8). Environmentally relevant concentrations of imidacloprid measured in airborne particulate matter and non-target soils pose a potential risk to coprophagous scarabs.

Effects of the neonicotinoid insecticides thiamethoxam and imidacloprid on metamorphosis of the toad Rhinella arenarum at environmentally-relevant concentrations.

The goal of the present study was to examine the effects of environmentally relevant concentrations of the neonicotinoid insecticides thiamethoxam and imidacloprid on the metamorphosis of the toad Rhinella arenarum. Tadpoles were exposed from stage 27 until completion of metamorphosis to concentrations of thiamethoxam ranging between 1.05 and 1050 µg/L and concentrations of imidacloprid varying between 3.4 and 3400 µg/L. The two neonicotinoids were found to act differently at the range of concentrations tested. Thiamethoxam did not markedly alter the final % tadpoles completing metamorphosis but extended by 6-20 days the time needed for tadpoles to complete metamorphosis. The extra number of days required to reach metamorphosis was concentration-dependent between 1.05 and 100.5 µg/L, and then stable at 20 days between 100.5 and 1005 µg/L. In contrast, imidacloprid did not significantly interfere with the overall time needed to complete metamorphosis but decreased success of metamorphosis at 3400 µg/L, the highest concentration tested. Both neonicotinoid concentrations did not markedly alter body size and weight of the newly metamorphosed toads. With a lowest observed effect concentration (LOEC) of 1.05 µg/L, thiamethoxam may be more likely to impact tadpole development in the wild compared to imidacloprid, which was without any apparent effect at concentrations up to 340 µg/L (no-observed effect concentration or NOEC). As the influence of thiamethoxam was triggered after tadpoles had reached Stage 39, when metamorphosis is strictly dependent upon thyroid hormones, this observed effect is attributed to result from actions of this neonicotinoid insecticide on the hypothalamic-pituitary-thyroid axis.

Assessment of Global DNA Methylation in SH-SY5Y Cells Exposed to the Neonicotinoid Insecticides Imidacloprid and Thiamethoxam.

Neonicotinoid insecticides, known for their selectivity and low mammalian toxicity, have been widely used in recent years as alternatives to organophosphate insecticides. Although neonicotinoids are generally considered to be safe, data show that they can cause harmful effects on human and environmental health. Due to the lack of information on their mechanism of toxicity, the effects of imidacloprid and thiamethoxam on DNA methylation as the most used marker for epigenetic effects were investigated in human neuroblastoma (SH-SY5Y) cells. The cells were exposed to imidacloprid and thiamethoxam in concentrations of 100, 200, and 500 µM for 24 hours, then global DNA methylation and expression of genes involved in global DNA methylation (DNMT1, DNMT3a and DNMT3b) were investigated. Global DNA methylation significantly increased after imidacloprid exposure at 100 µM, and thiamethoxam exposures at 200 µM and 500 µM (>1.5-fold). Imidacloprid significantly decreased the expression of DNMT1 and DNMT3a, whereas thiamethoxam did not cause any significant changes in the expression of DNMT genes. Our findings suggested that alteration in global DNA methylation may be involved in the toxic mechanisms of imidacloprid and thiametoxam.

Acute Toxicity of a Novel anti-fouling Material Additive DCOIT to Marine Chlorella sp.

DCOIT (4,5-dichloro-2-n-octyl-4-isothiazolin-3-one) is the main ingredient in SeaNine-211, a new antifouling agent that replaces organotin compounds to prevent the growth of fouling organisms on board. Biocides from antifoulants can cause problems for marine ecosystems by destroying non-target algal species. This study evaluated the potential adverse effects DCOIT using the Marine Chlorella sp. The concentration of DCOIT were set according to the semi-inhibitory concentrations for acute exposure experiments, and relevant oxidative stress indicators were measured to assess the acute toxic effects. The results showed that the inhibition concentrations (IC50) of DCOIT against Marine Chlorella sp was 2.522 mg/L. The genes related to photosynthesis and antioxidant capacity showed the effect of promoting low concentration and inhibiting high concentration. In addition, based on the ultrastructural observation and the expression analysis of photosynthesis related genes, it was found that DCOIT had a significant effect on plant photosynthesis.

Potential COVID-19 Drug Candidates Based on Diazinyl-Thiazol-Imine Moieties: Synthesis and Greener Pastures Biological Study.

A novel series of 1-aryl-N-[4-phenyl-5-(arylazo)thiazol-2-yl)methanimines has been synthesized via the condensation of 2-amino-4-phenyl-5-arylazothiazole with various aromatic aldehydes. The synthesized imines were characterized by spectroscopic techniques, namely 1H and 13C-NMR, FTIR, MS, and Elemental Analysis. A molecular comparative docking study for 3a-f was calculated, with reference to two approved drugs, Molnupiravir and Remdesivir, using 7BQY (Mpro; PDB code 7BQY; resolution: 1.7 A°) under identical conditions. The binding scores against 7BQY were in the range of -7.7 to -8.7 kcal/mol for 3a-f. The high scores of the compounds indicated an enhanced binding affinity of the molecules to the receptor. This is due to the hydrophobic interactions and multi-hydrogen bonds between 3a-f ligands and the receptor's active amino acid residues. The main aim of using in silco molecular docking was to rank 3a-f with respect to the approved drugs, Molnupiravir and Remdesivir, using free energy methods as greener pastures. A further interesting comparison presented the laydown of the ligands before and after molecular docking. These results and other supporting statistical analyses suggested that ligands 3a-f deserve further investigation in the context of potential therapeutic agents for COVID-19. Free-cost, PASS, SwissADME, and Way2drug were used in this research paper to determine the possible biological activities and cytotoxicity of 3a-f.

Discovery of Novel Histone Deacetylase 6 (HDAC6) Inhibitors with Enhanced Antitumor Immunity of Anti-PD-L1 Immunotherapy in Melanoma.

A series of 2-phenylthiazole analogues were designed and synthesized as potential histone deacetylase 6 (HDAC6) inhibitors based on compound 12c (an HDAC6/tubulin dual inhibitor discovered by us recently) and CAY10603 (a known HDAC6 inhibitor). Among them, compound XP5 was the most potent HDAC6 inhibitor with an IC50 of 31 nM and excellent HDAC6 selectivity (SI = 338 for HDAC6 over HDAC3). XP5 also displayed high antiproliferative activity against various cancer cell lines including the HDACi-resistant YCC3/7 gastric cancer cells (IC50 = 0.16-2.31 µM), better than CAY10603. Further, XP5 (50 mg/kg) exhibited significant antitumor efficacy in a melanoma tumor model with a tumor growth inhibition (TGI) of 63% without apparent toxicity. Moreover, XP5 efficiently enhanced the in vivo antitumor immune response when combined with a small-molecule PD-L1 inhibitor, as demonstrated by the increased tumor-infiltrating lymphocytes and reduced PD-L1 expression levels. Taken together, the above results suggest that XP5 is a promising HDAC6 inhibitor deserving further investigation.

Toxic Effects of High-dose Meloxicam and Carprofen on Female CD1 Mice.

The nonsteroidal anti-inflammatory drugs meloxicam and carprofen are commonly used as analgesics in mice. The current recommended doses of meloxicam at 0.2-1.0 mg/kg once daily and carprofen at 5-10 mg/kg twice daily may not be adequate to provide analgesia in mice. Several studies have suggested that doses up to 20 mg/kg of meloxicam and carprofen are needed to provide analgesic efficacy. This study investigated the clinical safety of these higher doses of meloxicam and carprofen by evaluating their potential for renal and gastrointestinal toxicity. Female CD-1 mice were given 20 mg/kg of either meloxicam, carprofen, or an equivalent volume of saline subcutaneously once daily for 3 or 7 d. On day 4, mice treated for 3 d were euthanized, and on days 8 and 15, mice treated for 7 d were euthanized. Blood was collected by cardiocentesis for serum chemistry analysis. Feces was collected from the colon for fecal occult blood testing, and tissues were collected for histopathology. No clinically significant changes in serum chemistry profiles were found in the drug-treated mice at any time point as compared with the saline controls. Fecal occult blood and histologic evidence of gastritis was associated with meloxicam administration in mice evaluated at days 4 and 8. By day 15, there was no association with meloxicam treatment and the presence of fecal occult blood or gastritis. There was no association between fecal occult blood and gastritis in the carprofen or saline-treated mice regardless of the treatment durations. These findings suggest that 20 mg/kg of meloxicam in mice causes gastric toxicity when given for 3 or 7 d and should be used cautiously; however, carprofen at 20 mg/kg appears to have minimal toxic effects with regard to the parameters measured.

Juvenile African Clawed Frogs (Xenopus laevis) Express Growth, Metamorphosis, Mortality, Gene Expression, and Metabolic Changes When Exposed to Thiamethoxam and Clothianidin.

Neonicotinoids (NEO) represent the main class of insecticides currently in use, with thiamethoxam (THX) and clothianidin (CLO) primarily applied agriculturally. With few comprehensive studies having been performed with non-target amphibians, the aim was to investigate potential biomarker responses along an adverse outcome pathway of NEO exposure, whereby data were collected on multiple biological hierarchies. Juvenile African clawed frogs, Xenopus laevis, were exposed to commercial formulations of THX and CLO at high (100 ppm) and low (20 ppm) concentrations of the active ingredient. Mortality, growth, development, liver metabolic enzyme activity, and gene expression endpoints were quantified. Tadpoles (n > 1000) from NF 47 through tail resorption stage (NF 66) were exposed to NEO or to NEO-free media treatments. Liver cell reductase activity and cytotoxicity were quantified by flow cytometry. Compared to control reference gene expressions, levels of expression for NEO receptor subunits, cell structure, function, and decontamination processes were measured by RT-qPCR by using liver and brain. Mortality in THX high was 21.5% compared to the control (9.1%); the metabolic conversion of THX to CLO may explain these results. The NF 57 control tadpoles were heavier, longer, and more developed than the others. The progression of development from NF 57-66 was reduced by THX low, and weight gain was impaired. Liver reductases were highest in the control (84.1%), with low NEO exhibiting the greatest reductions; the greatest cytotoxicity was seen with THX high. More transcriptional activity was noted in brains than in livers. Results affirm the utility of a study approach that considers multiple complexities in ecotoxicological studies with non-target amphibians, underscoring the need for simultaneously considering NEO concentration-response relationships with both whole-organism and biomarker endpoints.

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.

Thifluzamide exposure induced neuro-endocrine disrupting effects in zebrafish (Danio rerio).

Thifluzamide is widely used fungicide and frequently detected in aquatic system. In this study, the toxicity of fungicide thifluzamide to non-targeted aquatic organisms was investigated for neuroendocrine disruption potentials. Here, zebrafish embryos were exposed to a series of concentrations of thifluzamide for 6 days. The results showed that both the development of embryos/larvae and the behavior of hatched larvae were significantly affected by thifluzamide. Importantly, the decreased activity of acetylcholinesterase (AchE) and the increased contents of neurotransmitters such as serotonin (5-HT) and norepinephrine (NE), along with transcriptional changes of nervous system related genes were observed following 4 days exposure to thifluzamide. Besides, the decreased contents of triiodothyronine (T3) and thyroxine (T4) in whole body, as well as significant expression alteration in hypothalamic-pituitary-thyroid (HPT) axis associated genes were discovered in zebrafish embryos after 4 days of exposure to thifluzamide. Our results clearly demonstrated that zebrafish embryos exposed to thifluzamide could disrupt neuroendocrine, compromise behavior and induce developmental abnormality, suggesting impact of this fungicide on developmental programming in zebrafish.

Enzymatic responses in the head and midgut of Africanized Apis mellifera contaminated with a sublethal concentration of thiamethoxam.

The increasing use of insecticides, promoted by the intensification of agriculture, has raised concerns about their influence on the decline of bee colonies, which play a fundamental role in pollination. Thus, it is fundamental to elucidate the effects of insecticides on bees. This study investigated the damage caused by a sublethal concentration of thiamethoxam - TMX (0.0227 ng/µL of feed) in the head and midgut of Africanized Apis mellifera, by analyzing the enzymatic biomarkers, oxidative stress, and occurrence of lipid peroxidation. The data showed that the insecticide increased acetylcholinesterase activity (AChE) and glutathione-S-transferase (GST), whereas carboxylesterase (CaE3) activity decreased in the heads. Our results indicate that the antioxidant enzymes were less active in the head because only glutathione peroxidase (GPX) showed alterations. In the midgut, there were no alkaline phosphatase (ALP) or superoxide dismutase (SOD) responses and a decrease in the activity of CaE was observed. Otherwise, there was an increase in GPX, and the TBARS (thiobarbituric acid reactive substances) assay also showed differences in the midgut. The TBARS (thiobarbituric acid reactive substances) assay also showed differences in the midgut. The results showed enzymes such as CaE3, GST, AChE, ALP, SOD, and GPX, as well as the TBARS assay, are useful biomarkers on bees. They may be used in combination as a promising tool for characterizing bee exposure to insecticides.

DCOIT unbalances the antioxidant defense system in juvenile and adults of the marine bivalve Amarilladesma mactroides (Mollusca: Bivalvia).

DCOIT is a co-biocide that is part of the formulation of the commercial antifouling Sea-Nine 211® and although it is "safe to use", negative effects have been reported on the antioxidant defense system of non-target organisms. Therefore, the objective of this research was to verify and compare the response of antioxidant enzymes of juveniles and adults of Amarilladesma mactroides exposed to DCOIT. The animals were exposed to solvent control (DMSO 0.01%) and DCOIT (measured concentration 0.01 mg/L and 0.13 mg/L) for 96 h, then gills, digestive gland and mantle were collected for analysis of the enzymatic activity of glutathione S-transferase (GST), superoxide dismutase (SOD) and catalase (CAT). The results revealed that adults, in relation to juveniles, have low basal activity of GST and SOD enzymes in the gills and digestive gland and high basal activity of SOD and CAT in the mantle. DCOIT did not alter GST activity in the gills of any life stage, while both concentrations decreased SOD and CAT in adults. In the digestive gland, it was observed that DCOIT (0.13 mg/L) decreased the GST activity in adults and CAT in juveniles, and both concentrations of the co-biocide decreased the SOD and CAT in adults. In the mantle, DCOIT (0.13 mg/L) increased CAT in juveniles. We conclude that juveniles have greater basal activity of antioxidant enzymes than adults and, in addition, DCOIT negatively affected the adults of A. mactroides, mainly decreasing the activity of GST, SOD and CAT in the gills and digestive gland of these organisms.

Higher oral efficacy of ravuconazole in self-nanoemulsifying systems in shorter treatment in experimental chagas disease.

We investigated the in vitro activity and selectivity, and in vivo efficacy of ravuconazole (RAV) in self-nanoemulsifying delivery system (SNEDDS) against Trypanosoma cruzi. Novel formulations of this poorly soluble C14-α-demethylase inhibitor may improve its efficacy in the experimental treatment. In vitro activity was determined in infected cardiomyocytes and efficacy in vivo evaluated in terms of parasitological cure induced in Y and Colombian strains of T. cruzi-infected mice. In vitro RAV-SNEDDS exhibited significantly higher potency of 1.9-fold at the IC50 level and 2-fold at IC90 level than free-RAV. No difference in activity with Colombian strain was observed in vitro. Oral treatment with a daily dose of 20 mg/kg for 30 days resulted in 70% of cure for RAV-SNEDDS versus 40% for free-RAV and 50% for 100 mg/kg benznidazole in acute infection (T. cruzi Y strain). Long-term treatment efficacy (40 days) was able to cure 100% of Y strain-infected animals with both RAV preparations. Longer treatment time was also efficient to increase the cure rate with benznidazole (Y and Colombian strains). RAV-SNEDDS shows greater efficacy in a shorter time treatment regimen, it is safe and could be a promising formulation to be evaluated in other pre-clinical models to treat T. cruzi and fungi infections.

Amended Safety Assessment of Methylchloroisothiazolinone and Methylisothiazolinone as Used in Cosmetics.

The Expert Panel for Cosmetic Ingredient Safety (Panel) reassessed the safety of the mixture Methylchloroisothiazolinone (MCI)/Methylisothiazolinone (MI), which functions as a preservative in cosmetic products. The Panel reviewed relevant animal and human data provided in this safety assessment, and data from the previously published safety assessment of this mixture, and concluded that MCI/MI is safe in cosmetics when formulated to be nonsensitizing, based on the results of a quantitative risk assessment or similar methodology; however, at no point should concentrations exceed 7.5 ppm in leave-on products or 15 ppm in rinse-off products.

Amended Safety Assessment of Methylisothiazolinone as Used in Cosmetics.

The Expert Panel for Cosmetic Ingredient Safety (Panel) reassessed the safety of Methylisothiazolinone, which functions as a preservative in cosmetics. The Panel reviewed relevant animal and human data provided in this safety assessment, and data from the previously published safety assessments of Methylisothiazolinone, and concluded that Methylisothiazolinone is safe for use in rinse-off cosmetic products at concentrations up to 100 ppm (ie, 0.01%) and safe in leave-on cosmetic products when they are formulated to be nonsensitizing, which may be determined based on a quantitative risk assessment or similar methodology.

Comparative study of two isothiazolinone biocides, 1,2-benzisothiazolin-3-one (BIT) and 4,5-dichloro-2-n-octyl-isothiazolin-3-one (DCOIT), on barrier function and mitochondrial bioenergetics using murine brain endothelial cell line (bEND.3).

Isothiazolinone (IT) biocides are potent antibacterial substances used as preservatives and disinfectants. These biocides exert differing biocidal effects and display environmental stability based upon chemical structure. In agreement with our recent study reporting that 2-n-octyl-4-isothiazolin-3-one (OIT) induced dysfunction of the blood-brain barrier (BBB), the potential adverse health effects of two IT biocides 1,2-benzisothiazolin-3-one (BIT) and 4,5-dichloro-2-n-octyl-isothiazolin-3-one (DCOIT) were compared using brain endothelial cells (ECs) derived from murine brain endothelial cell line (bEND.3). BIT possesses an unchlorinated IT ring structure and used as a preservative in cleaning products. DCOIT contains a chlorinated IT ring structure and employed as an antifouling agent in paints. Data demonstrated that DCOIT altered cellular metabolism at a lower concentration than BIT. Both BIT and DCOIT increased reactive oxygen species (ROS) generation at the mitochondrial and cellular levels. However, the effect of DCOIT on glutathione (GSH) levels appeared to be greater than BIT. While mitochondrial membrane potential (MMP) was decreased in both BIT- and DCOIT-exposed cells, direct disturbance in mitochondrial bioenergetic flux was only observed in BIT-treated ECs. Taken together, IT biocides produced toxicity in brain EC and barrier dysfunction, but at different concentration ranges suggesting distinct differing mechanisms related to chemical structure.