Multiple biomarkers in pufferfish as a proxy of environmental health in brazilian marine protected areas.

Marine Protected Areas (MPAs) are designed to conserve biodiversity and vulnerable ecosystems. This study aimed to assess the environmental quality of three Brazilian MPAs, based on the integrated analysis of biomarkers in pufferfish. The MPAs are differentiated by the degree of anthropogenic influences. The Barra do Una Estuary sustainable reserve (JUR) is a reference area due to its low levels of contamination and species diversity. The Cananéia Estuarine System (CAN) has been recognized as a biosphere reserve by UNESCO, as well as a Ramsar wetland. This MPA was influenced by upstream mining activities, resulting in the introduction of metals in the estuarine waters and the discharge of untreated urban sewage. The São Vicente estuary (SSV) lacks proper sanitation infrastructure. All collections were conducted during winter season, and, after collection, the animals were euthanized, their soft tissues were removed, and multiple biomarkers were analyzed in the gills and liver, as biometric, genotoxic, biochemical, and morphological. A one-factor multivariate analysis was applied to evaluate the differences between the data sets, and the matrices were analyzed using PERMANOVA to evaluate the "estuary" factor. The results were integrated using PCA with a 0.4 cut value and an Enhanced Integrated Biomarker Response (EIBR) was calculated. PCA was correlated with biochemical, genotoxic, and morphological biomarkers. In general, SVV differed from CAN and JUR as shown by both univariate and multivariate analyses. SVV also showed the highest EIBR, followed by CAN. Organisms from SSV showed greater gill pathology, elevated AChE activity and lipoperoxidation (LPO), and micronuclei frequency. CAN present intermediate EIBR, with severe pathologies in the liver. CAN seems to present an intermediate environmental quality between SSV and JUR indicating the importance of the existence of MPAs for environmental conservation and the need to monitor such areas, to maintain their suitable environmental quality.

Therapeutic Activity of a Topical Formulation Containing 8-Hydroxyquinoline for Cutaneous Leishmaniasis.

Cutaneous leishmaniasis exhibits a wide spectrum of clinical manifestations; however, only a limited number of drugs are available and include Glucantime® and amphotericin B, which induce unacceptable side effects in patients, limiting their use. Thus, there is an urgent demand to develop a treatment for leishmaniasis. Recently, it was demonstrated that 8-hydroxyquinoline (8-HQ) showed significant leishmanicidal effects in vitro and in vivo. Based on that, this work aimed to develop a topical formulation containing 8-HQ and assess its activity in experimental cutaneous leishmaniasis. 8-HQ was formulated using a Beeler base at 1 and 2% and showed an emulsion size with a D50 of 25 and 51.3 µm, respectively, with a shear-thinning rheological behaviour. The creams were able to permeate artificial Strat-M membranes and excised porcine skin without causing any morphological changes in the porcine skin or murine skin tested. In BALB/c mice infected with L. (L.) amazonensis, topical treatment with creams containing 1 or 2% of 8-HQ was found to reduce the parasite burden and lesion size compared to infected controls with comparable efficacy to Glucantime® (50 mg/kg) administered at the site of the cutaneous lesion. In the histological section of the skin from infected controls, a diffuse inflammatory infiltrate with many heavily infected macrophages that were associated with areas of necrosis was observed. On the other hand, animals treated with both creams showed only moderate inflammatory infiltrate, characterised by few infected macrophages, while tissue necrosis was not observed. These histological characteristics in topically treated animals were associated with an increase in the amount of IFN-γ and a reduction in IL-4 levels. The topical use of 8-HQ was active in decreasing tissue parasitism and should therefore be considered an interesting alternative directed to the treatment of leishmaniasis, considering that this type of treatment is non-invasive, painless, and, importantly, does not require hospitalisation, improving patient compliance by allowing the treatment to be conducted.

Evaluation of myenteric neurons in the colon of rats exposed to 2,4 dichlorophenoxyacetic acid herbicide.

The assessment of the enteric nervous system provides a better understanding of the effects that contaminants can have on the health and well-being of organisms. It has been reported that 2,4-dichlorophenoxyacetic acid (2,4-D) is a highly persistent herbicide in the environment that is responsible for neurotoxic changes in different myenteric neuronal subpopulations. The current study aimed to evaluate the effects of 2,4-D on myenteric neurons in the colon of Rattus norvegicus for the first time. A dose of 2,4-D (5 mg/kg/day) was administered to the experimental group (2,4-D) for 15 days. Then, the proximal colon was collected and submitted to Giemsa and NADPH-d histochemical techniques for the disclosure of total and nitrergic neurons. The 2,4-D group presented a higher density of total neurons (p = 0.05, t-test), which together with the maintenance of nitrergic neuronal density, may be related to the increase in the expression of the neurotransmitter acetylcholine by colocalization, responsible for stimulating the intestinal smooth muscle and increasing the chances of the expulsion of the harmful content present in the lumen. Over 15 days, the neurotoxic effects of 2,4-D in the myenteric plexus influenced an increase in the general population of myenteric neurons in the colon.

Integrated analysis of fish intestine biomarkers: Complementary tools for pollution assessment.

The gastrointestinal tract and its enteric nervous system are the first routes of food and xenobiotics uptake. Considering the importance of this organ, this study evaluated intestinal biomarkers of Sphoeroides testudineus integrating the data to generate tools for pollution assessment. The fish were collected in three sites of São Paulo Coast and their intestines were analyzed for biochemical, histology, and neuronal density and morphometry biomarkers. To evaluate the differences among the data, a PERMANOVA was applied, followed by a FA/PCA. The PERMANOVA indicated differences (P < 0.001) between the regions (RA, A1, and A2). Four factors were extracted from the FA/PCA (62% cumulative), showing that the animals from A2 presented severe alterations, mainly in intestinal morphometry and neuronal density. A1 alterations refer mainly to the increase of neuronal metabolism. Our results also evidence a gradient of environmental quality related to the protection level (AR > A1 > A2).

A preliminary study on multi-level biomarkers response of the tropical oyster Crassostrea brasiliana to exposure to the antifouling biocide DCOIT.

This study investigated the sublethal effects of environmentally relevant concentrations of DCOIT on the neotropical oyster Crassostrea brasiliana. Gills and digestive glands of animals exposed to increasing concentrations of DCOIT were analyzed for biochemical, cellular, and histopathological responses. Exposure to DCOIT (0.2 to 151 µg L-1) for 120 h triggered oxidative stress in both tissues (through the modulation of GPX, GST, GSH and GR), which led to damage of membrane lipids (increase of LPO and reduction of the NRRT). DCOIT increased histopathological pathologies in gills, such as necrosis, lymphocyte infiltration and epithelial desquamation. This study showed that short term exposure to environmental concentrations of DCOIT causes negative effects on C. brasiliana at biochemical, physiological, and histological levels. Therefore, the use of DCOIT as a booster biocide in antifouling paints should be further assessed, as it may cause environmental hazards to marine organisms.

Adaptative responses of myenteric neurons of Sphoeroides testudineus to environmental pollution.

Contamination in estuarine regions affects the local biota damaging the ecosystems and reaching humans. The gastrointestinal tract is a dynamic environment capable of obtaining nutrients and energy from food while it protects the host against harmful toxins and pathogens from the external environment. These functions are modulated by the enteric nervous system and changes in its structure can result in gastrointestinal disorders. The objective of this study was to evaluate if the environmental contaminants have effects on the myenteric neuronal plasticity of pufferfish Sphoeroides testudineus. Animals were collected in Barra do Una River, located at Jureia-Itatins Mosaic of Protected Areas (reference area - RA) and in the Santos Estuarine System (impacted area - IA). Morpho-quantitative analyses of the general and metabolically active myenteric neuronal populations of the proximal and distal intestine were made. Disarrangement was observed in the general organization of the myenteric plexus, with an expressive reduction of the neuronal groups (nodes) in the animals of IA. The vulnerability of the myenteric plexus was evidenced by a decrease in density and cellular profile of the general neuronal population, followed by an increase of the metabolism of the remaining neurons, which in turn was verified by a growth of the area of the cellular and nuclear profiles of the metabolically active neuronal population. Through these analyses, we concluded that animals inhabiting polluted regions present alterations in the myenteric neuronal plasticity, as a way of maintaining the functions of the gastrointestinal tract.

Balanced Caloric Restriction Minimizes Changes Caused by Aging on the Colonic Myenteric Plexus.

Aging can promote significant morphofunctional changes in the gastrointestinal tract (GIT). Regulation of GIT motility is mainly controlled by the myenteric neurons of the enteric nervous system. Actions that aim at decreasing the aging effects in the GIT include those related to diet, with caloric restriction (CR). The CR is achieved by controlling the amount of food or by manipulating the components of the diet. Therefore, the objective of this study was to evaluate different levels of CR on the plasticity of nicotinamide adenine dinucleotide phosphate- (NADPH-) reactive myenteric neurons in the colon of Wistar rats during the aging process using ultrastructural (transmission electron microscopy) and morphoquantitative analysis. Wistar male rats (Rattus norvegicus) were distributed into 4 groups (n = 10/group): C, 6-month-old animals; SR, 18-month-old animals fed a normal diet; CRI, 18-month-old animals fed a 12% CR diet; CRII, 18-month-old animals fed a 31% CR diet. At 6 months of age, animals were transferred to the laboratory animal facility, where they remained until 18 months of age. Animals of the CRI and CRII groups were submitted to CR for 6 months. In the ultrastructural analysis, a disorganization of the periganglionar matrix with the aging was observed, and this characteristic was not observed in the animals that received hypocaloric diet. It was observed that the restriction of 12.5% and 31% of calories in the diet minimized the increase in density and cell profile of the reactive NADPH neurons, increased with age. This type of diet may be adapted against gastrointestinal disturbances that commonly affect aging individuals.