Isabella Fernandes Delgado

Intoxicações e uso de pesticidas por agricultores do Município de Paty do Alferes, Rio de Janeiro, Brasil

This survey is part of a more comprehensive study on the health consequences of pesticide exposure. In the county (municipality) of Paty do Alferes, Rio de Janeiro State, Brazil, 55 agricultural workers were interviewed on the use of pesticides, use of personal protective equipment, data on health status, and symptoms related to pesticide exposure, disposal of agrochemical containers, and technical assistance. The most widely used pesticides were insecticides such as abamectin, organophosphate compounds, and pyrethroids, and fungicides such as mancozeb, chlorothalonil, and copper products. As a rule, pesticides are handled carelessly, and 92% of workers involved in the mixing, loading, and spraying of insecticides and fungicides used no protective clothing or equipment whatsoever. Some 62% of workers reported at least one illness associated with mixing or spraying pesticides. The most frequently reported symptoms were headache, nausea, vomiting, dizziness, skin irritation, and blurred vision, and 21% of affected workers required medical care. In more than half (51%) of the cases, workers reported using organophosphate insecticides from toxicological class I when they felt sick.

Chlorinated dibenzo-p-dioxins and dibenzofurans and the human immune system. 1. Blood cell receptors in volunteers with moderately increased body burdens

Using monoclonal antibodies (mAbs) and flow cytometry, we studied a variety of surface receptors on lymphocyte subpopulations of workers with moderately increased body burdens of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and of other polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/PCDF), expressed here as International-Toxicity Equivalencies (I-TE). The hypothesis to be tested was whether or not humans exhibit a similar susceptibility to PCDDs/PCDFs with respect to the surface receptors found previously to respond to small doses of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in Callithrix jacchus. These are: helper-inducer (memory) T cells (CD4+CD45R0+CD45RA-CD29highCD11a+), CD20+ B cells, and cytotoxic T cells (CD8+CD56+/CD57+). Furthermore, 68 triple-labellings with mAbs were performed on the cells of each volunteer to possibly generate further hypotheses. It was evaluated whether any of the variables might be used as a biomarker of effects for this class of compounds. There were two main goals: (1) to evaluate whether workers with a moderately increased PCDD/PCDF-body burden [25-140 ppt TCDD or 104-522 ppt I-TE in blood fat] exhibit changes in the surface receptors of white blood cells, as observed in previous studies in non-human primates, and (2) to clarify whether persons at the upper range [10-23 ppt TCDD or 30-90 ppt I-TE in blood fat] of the body burden reference values of a not particularly exposed population show detectable deviations in these immunological variables, when compared with persons at the lower and medium range [1-3 ppt TCDD or 9-29 ppt I-TE] of these body burden reference values. Regression analysis of our data revealed slight trends for some of the biomarkers (e.g. CD45R0+). With one exception, these were all increases. None of the alterations observed are of medical relevance. The slight increase in the percentage of CD4+CD45R0+ cells remained significant even after covariant analysis taking age-related changes into account. Altogether, the data do not provide any evidence to support an assumption that moderately increased body burdens of PCDDs/PCDFs in adults induce decreases in the cellular components of the human immune system. Adult humans certainly are less susceptible to this action of PCDDs/PCDFs than adolescent Callithrix jacchus.

Study on embryo-foetotoxicity of β-myrcene in the rat

beta-Myrcene is a constituent of many essential oils that have been used extensively in cosmetic fragrances and as flavouring additives in the food industry. Recently, this monoterpene was reported to be an analgesic substance. Notwithstanding the widespread use of myrcene and essential oils containing myrcene in perfume and in food additives, experimental studies on the toxicity of this substance are still scarce. This study aimed to provide data on the embryo-foetotoxic potential of beta-myrcene in the rat. beta-Myrcene (0.25, 0.5 and 1.2 g/kg) in corn oil was given orally to Wistar rats from day 6 to 15 of pregnancy. Caesarean sections were performed on day 20 of pregnancy, and the number of resorptions and implantation sites were recorded. Foetuses were weighed, examined for external malformations, and fixed for visceral examination, or cleared and stained with Alizarin Red S for skeleton evaluation. No adverse effects were seen with the two lowest doses tested. Decreased weight gain during the first days of treatment and the death of one of 29 treated dams indicated that the highest dose tested (1.2 g/kg) induced maternal toxicity. A higher incidence of signs of retardation and of anomalies in the foetal skeleton indicated that 1.2 g/kg was also toxic to the rat embryo. From the data presented in this paper the no-observed-adverse-effect level for embryo-foetotoxicity could be set at 0.5 g beta-myrcene/kg body weight.

Postnatal Development and Fertility of Offspring from Mice Exposed to Triphenyltin (Fentin) Hydroxide During Pregnancy and Lactation

Fentin or triphenylthin (TPT) is an organotin compound (OTC) widely used as an agricultural fungicide and miticide. It is well known that TPT exerts adverse effects on the reproductive and immune systems and may disrupt the endocrine system, raising concerns regarding the risks posed by exposure to this metal on environmental and human health. In this study the effects of maternal exposure to TPT at doses of control (0), 1.875, 3.75, or 7.5 mg/kg body weight/d, po, were examined during gestation and lactation on offspring growth, organ weights, and fertility. Except for a significant liver enlargement at the highest dose, TPT produced no maternal toxicity. Increased neonatal mortality (death of 3 entire litters from a total of 18 treated litters) was noted at 7.5 mg/kg. Pup body weight at birth was significantly reduced at all dose levels, but no marked weight loss was found on postnatal day (PND) 5 and thereafter. Offspring maturation (ear unfolding, incisor eruption, vagina opening, and testes descent) and fertility in adulthood were not significantly affected by maternal exposure to TPT. In conclusion, data provided by this study indicate that maternal treatment with TPT during pregnancy and lactation delayed prenatal growth but did not impair postnatal development and fertility in exposed offspring in adulthood.

Standardisation of defined approaches for skin sensitisation testing to support regulatory use and international adoption: position of the International Cooperation on Alternative Test Methods

Skin sensitisation is the regulatory endpoint that has been at the centre of concerted efforts to replace animal testing in recent years, as demonstrated by the Organisation for Economic Co-operation and Development (OECD) adoption of five non-animal methods addressing mechanisms under the first three key events of the skin sensitisation adverse outcome pathway. Nevertheless, the currently adopted methods, when used in isolation, are not sufficient to fulfil regulatory requirements on the skin sensitisation potential and potency of chemicals comparable to that provided by the regulatory animal tests. For this reason, a number of defined approaches integrating data from these methods with other relevant information have been proposed and documented by the OECD. With the aim to further enhance regulatory consideration and adoption of defined approaches, the European Union Reference Laboratory for Alternatives to Animal testing in collaboration with the International Cooperation on Alternative Test Methods hosted, on 4–5 October 2016, a workshop on the international regulatory applicability and acceptance of alternative non-animal approaches, i.e., defined approaches, to skin sensitisation assessment of chemicals used in a variety of sectors. The workshop convened representatives from more than 20 regulatory authorities from the European Union, United States, Canada, Japan, South Korea, Brazil and China. There was a general consensus among the workshop participants that to maximise global regulatory acceptance of data generated with defined approaches, international harmonisation and standardisation are needed. Potential assessment criteria were defined for a systematic evaluation of existing defined approaches that would facilitate their translation into international standards, e.g., into a performance-based Test Guideline. Informed by the discussions at the workshop, the ICATM members propose practical ways to further promote the regulatory use and facilitate adoption of defined approaches for skin sensitisation assessments.

Determination of eye irritation potential of low-irritant products: comparison of in vitro results with the in vivo draize rabbit test

In an attempt to build the evaluation strategies to assess the human eye irritation, a reassessment of some in vitro tests is necessary, particularly concerning the non-irritants, mild and moderate irritants. Thus, the correlations between results obtained from the Draize test with the in vitro methods HET-CAM (Hens Egg Test-Chorion Allantonic Membrane) and RBC haemolysis assay to assess the ocular irritancy potential of 20 eye drops were examined. Parameters such as accuracy (%) and specificity (%) were determined. All results were correlated with RBC correctly with the results obtained in the Draize test. The HET-CAM presented four false-positive results, showing a tendency of data overestimation. Despite the high specificity provided mainly by the RBC, it would be necessary to test a wider range of products representing all the scales of irritation to confirm its ability to be used as a first alternative to test products that could be presumptive non-irritants.

Postnatal development and resistance to Plasmodium yoelii infection of mice prenatally exposed to triphenyltin hydroxide.

In this study, we evaluated the effects of prenatal exposure to triphenyltin hydroxide (TPTH) on the postnatal development of Swiss Webster mice. Females were treated by gavage (0, 7.5 15 and 30 mg TPTH/kg/day) on days 6–17 of gestation. After birth, the progeny was examined for deaths, body weight gain and appearance of developmental landmarks. On postnatal day 50, one male and one female of each litter were inoculated with Plasmodium yoelii and the time‐course of infection was monitored. TPTH was embryolethal at doses ≥15 mg/kg/day. Body weight at birth was decreased, but no alteration of pup body weight was observed after postnatal day 5. Except for an advancement of incisor eruption in the group treated with 15 mg/kg/day, no alteration of somatic development was noted. A shorter latency to peak parasitemia and a reduced malaria‐induced spleen enlargement were observed in mice prenatally exposed to TPTH. In conclusion, prenatal exposure to TPTH at doses ≥15 mg/kg enhanced neonatal lethality, reduced pup birth weight and interfered with the response to infection with P. yoelii in adulthood.

Sexual maturation and fertility of mice exposed to triphenyltin during prepubertal and pubertal periods

Highlights • NOAEL for general toxicity was 1.875 (male) or 3.75 (female) mg of TPT/kg bw/d po.• Reproductive toxicity occurred only at doses of TPT that also caused general toxicity.• NOAEL for harmful effects on male reproduction was 1.875 mg of TPT/kg bw/d po.• NOAEL for harmful effects on female reproduction was 3.75 mg of TPT/kg bw/d po.• TPT-caused decrease in spermatid and sperm count was reversed after treatment.• Exposure to TPT during pre/pubertal period did not impair adult mice fertility.

International regulatory requirements for skin sensitization testing

Skin sensitization test data are required or considered by chemical regulation authorities around the world. These data are used to develop product hazard labeling for the protection of consumers or workers and to assess risks from exposure to skin-sensitizing chemicals. To identify opportunities for regulatory uses of non-animal replacements for skin sensitization tests, the needs and uses for skin sensitization test data must first be clarified. Thus, we reviewed skin sensitization testing requirements for seven countries or regions that are represented in the International Cooperation on Alternative Test Methods (ICATM). We noted the type of skin sensitization data required for each chemical sector and whether these data were used in a hazard classification, potency classification, or risk assessment context; the preferred tests; and whether alternative non-animal tests were acceptable. An understanding of national and regional regulatory requirements for skin sensitization testing will inform the development of ICATMs international strategy for the acceptance and implementation of non-animal alternatives to assess the health hazards and risks associated with potential skin sensitizers.

Applicability of the Monocyte Activation Test (MAT) for hyperimmune sera in the routine of the quality control laboratory : Comparison with the Rabbit Pyrogen Test (RPT)

Pyrogen tests are safety assays performed during the routine quality control of injectable products required by regulatory agencies. Currently, there are three available testing possibilities: 1) the Rabbit Pyrogen Test (RPT); 2) the Bacterial Endotoxin Test (BET); and 3) test systems using human whole-blood or monocytes, termed Monocyte Activation Test (MAT). Although BET is often considered as a replacement for the animal test, it is unable to detect pyrogens other than endotoxin. MAT is based on the human fever reaction and thus, most closely reflects the human response. The aim of this study was to conduct a parallel comparison of the RPT and MAT for hyperimmune sera (HS) batches analyzed during the routine of a quality control laboratory. MAT was performed in the same 43 batches of HS previously tested using RPT. The results showed that MAT presented 100% sensitivity and approximately 85% specificity as compared to RPT, i.e., no false-negative results were obtained. Few suspicious samples, which were negative in the RPT after retesting, provided divergent positive results suggesting a lower limit of detection of MAT. MAT is thus able to detect contaminants in biological products such as HS batches.