Are Microfibers a Threat to Marine Invertebrates? A Sea Urchin Toxicity Assessment.

The rise of "fast fashion" has driven up the production of low-cost, short-lived clothing, significantly increasing global textile fiber production and, consequently, exacerbating environmental pollution. This study investigated the ecotoxicological effects of different types of anthropogenic microfibers-cotton, polyester, and mixed fibers (50% cotton: 50% polyester)-on marine organisms, specifically sea urchin embryos. All tested fibers exhibited toxicity, with cotton fibers causing notable effects on embryonic development even at environmentally relevant concentrations. The research also simulated a scenario where microfibers were immersed in seawater for 30 days to assess changes in toxicity over time. The results showed that the toxicity of microfibers increased with both concentration and exposure duration, with polyester being the most toxic among the fibers tested. Although synthetic fibers have been the primary focus of previous research, this study highlights that natural fibers like cotton, which are often overlooked, can also be toxic due to the presence of harmful additives. These natural fibers, despite decomposing faster than synthetic ones, can persist in aquatic environments for extended periods. The findings underline the critical need for further research on both natural and synthetic microfibers to understand their environmental impact and potential threats to marine ecosystems and sea urchin populations.

Exposure to microplastics contaminated with pharmaceuticals and personal care products: Histological effects on Ucides cordatus.

Pharmaceuticals and personal care products (PPCPs) are known to interact with microplastics (MPs) in aquatic environments, with substances such as the antimicrobial triclosan (TCS) and the synthetic hormone 17α-ethinylestradiol (EE2) being prevalent. These persistent contaminants are linked to toxic effects in aquatic organisms. This study aimed to investigate histological and morphometric changes in the gills of Ucides cordatus exposed to microplastics alone and microplastics contaminated with PPCPs. The experimental design included four treatment groups: 1) control (C), 2) virgin microplastics (MP), 3) microplastics fortified with triclosan (MPT), and 4) microplastics fortified with 17α-ethinylestradiol (MPE), with exposure durations of 3 or 7 days. Significant differences were observed in the histopathological indices for treatments with PPCP-fortified microplastics at 3 days (MPT and MPE) and 7 days (MPT). Notable pathologies included necrosis, fibrosis, and circulatory disorders. Exposure duration was significantly associated with morphometric changes, including secondary lamellar width in MPT and secondary lamellar length in MPE. These findings indicate that exposure to microplastics contaminated with PPCPs may impair the osmoregulatory and respiratory functions of Ucides cordatus.

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.

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.

Exercise reduces inhibitory neuroactivity and protects myenteric neurons from age-related neurodegeneration.

The practice of regular exercise is indicated to prevent some motility disturbances in the gastrointestinal tract, such as constipation, during aging. The motility alterations are intimately linked with its innervations. The goal of this study is to determine whether a program of exercise (running on the treadmill), during 6 months, has effects in the myenteric neurons (NADH- and NADPH-diaphorase stained neurons) in the colon of rats during aging. Male Wistar rats 6 months (adult) and 12 months (middle-aged) old were divided into 3 different groups: AS (adult sedentary), MS (middle-aged sedentary) and MT (middle-aged submitted to physical activity). The aging did not cause a decline significant (p>0.05) of the number of NADH-diaphorase stained neurons in sedentary rats (AS vs. MS group). In contrast, a decline of 31% was observed to NADPH-diaphorase stained neurons. Thus, animals that underwent physical activity (AS vs. MT group) rescued neurons from degeneration caused by aging (total number, density and profile of neurons did not change with age--NADH-diaphorase method). On the other hand, physical activity augmented the decline of NADPH-diaphorase positive neurons (total number, density and profile of neurons decreased). Collectively, the results show that exercise inhibits age-related decline of myenteric neurons however, exercise augments the decline of neurons with inhibitory activity (nitric oxide) in the colon of the rats.

Enteric nervous system analyses: New biomarkers for environmental quality assessment.

The gastrointestinal tract (GIT) of fish is a target of contaminants since it can absorb these substances. We evaluated the morphophysiological alterations in the GIT of Sphoeroides testudineus collected in two estuaries presenting differences in their environmental quality (NIA and IA). The intestine was analyzed for histological and neuronal changes; liver and gills for biochemical markers; muscle tissues for neurotoxicity and peripheral blood for genotoxic damage. The results showed alterations in the GIT of the animals collected in the IA, such as muscle tunica and goblet cell density reduction, increased intraepithelial lymphocytes density and changes in neuronal density. Furthermore, changes were observed in MTs and LPO in the gills. Thus, we suggest that TGI is functioning as a barrier that responds to ingested contaminants, in order to reduce their absorption and translocation. Thus, alterations in morphophysiological and enteric neurons in S. testudineus can be used as biomarkers of environmental contamination.

Effect of caloric restriction on myenteric neuroplasticity in the rat duodenum during aging.

The objective of this study was to evaluate the effects of caloric restriction (CR) on myenteric neurons in the duodenum of Wistar rats during aging. Thirty rats were divided into three groups: the C group (six-month-old animals that were fed a normal diet from weaning until six months of age), the SR group (18-month-old animals that were fed a normal diet from weaning until 18 months of age) and the CR group (18-month-old animals that were fed a 30% CR diet after six months of age). After 12 months, the animals were euthanized. Whole-mount preparations of the duodenums were either stained with Giemsa or underwent NADPH-diaphorase histochemistry to determine the general myenteric neuron population and the nitrergic neuron subpopulation (NADPH-d+), respectively. The NADPH-d-negative (NADPH-d-) neuron population was estimated based on the difference between the Giemsa-stained and NADPH-d+ neurons. The neurons were counted, and the cell body areas were measured. Aging was associated with neuronal loss in the SR group, which was minimized by caloric restriction in the CR group. The density (mm(2)) of the Giemsa-stained neurons was higher in the SR group (79.09 ± 6.25) than in the CR (92.37 ± 11.6) and C (111.68 ± 15.26) groups. The density of the NADPH-d+ neurons was higher in the SR group (44.90 ± 5.88) than in the C (35.75 ± 1.6) and RC (39.14 ± 7.02) groups. The density of NADPH-d- neurons was higher in the CR (49.73 ± 12.08) and C (75.64 ± 17.05) groups than in the SR group (33.82 ± 4.5). In the C group, 32% and 68% of the Giemsa-stained myenteric neurons were NADPH-d+ or NADPH-d-, respectively. With aging (SR group), the percentage of nitrergic neurons (56.77%) increased, whereas the percentage of NADPH-d- neurons (43.22%) decreased. In the CR group, the change in the percentage of nitrergic (42.37%) and NADPH-d- (57.62%) neurons was lower. As NADPH-d- neurons will be mostly cholinergic neurons, CR appears to reduce the loss of cholinergic neurons during aging. The cell body dimensions (µm(2)) were not altered by aging or CR. Thus, CR had a protective effect on myenteric neurons during aging.

Morpho-quantitative study of NADH-diaphorase positive myenteric neurons of the ileum of rats of Holtzman lineage (Rattus norvegicus) - DOI: 10.4025/actascibiolsci.v29i3.472 / Estudo morfoquantitativo dos neurônios mioentéricos NADH-diaforase positivos do íleo de ratos (Rattus norvegicus) da linhagem Holtzman - DOI: 10.4025/actascibiolsci.v29i3.472

The myenteric plexus has a regular characteristic morphological pattern for each segment of the digestive tube and for each species of animal. Considering the lack of data pertaining to the mentioned plexus in rats of Holtzman lineage, the objective of this investigation was to carry out a morpho-quantitative study of the myenteric neurons in the ileum, by means of histological sections and whole-mount muscular preparations treated by the NADH-diaphorase method. The profiles of the cell bodies (CB) of the neurons in the mesenteric and antimesenteric regions were counted and measured. The neurons were classified according to the dimensions of the CBs. NADH-dp myenteric neurons were observed grouped together into ganglia in the muscular tunica. The mean neuronal density was 985.8 ± 195.4 neurons/8.96 mm2 in the antimesenteric region and 1267.8 ± 259.92 neurons/8.96 mm2 in the mesenteric region. The incidences of small, medium and large neurons were 14.4, 82 and 3.6% in the antimesenteric region and 14.6, 70.8 and 14.4% in the mesenteric region, respectively. It was concluded that ganglionated arrangements and medium-sized NADH-dp neurons predominated in the myenteric plexus of adult Holtzman rats. The results observed indicated that the NADH-dp myenteric neurons of the ileum of Holtzman rats are similar to those of rats of Wistar lineage with respect to their localization, gangli

O plexo mioentérico possui um padrão morfológico regular característico para cada segmento do tubo digestório e para cada espécie animal. Considerando a escassez de dados pertinentes ao referido plexo em ratos da linhagem Holtzman, o presente estudo como objetivo o estudo morfoquantitativo dos neurônios mioentéricos do íleo, por meio de cortes histológicos e preparados de membrana, tratados pelo método da NADH-diaforase. Foram contados e mensurados os perfis do corpo celular (PC) de neurônios nas regiões mesentérica e antimesentérica. Os neurônios foram classificados segundo as dimensões do PC. Foram observados os neurônios mioentéricos NADH-dp, reunidos em gânglios na túnica muscular. A densidade neuronal média foi de 985,8 ± 195,4 neurônios/8,96 mm2 na região antimesentérica e 1267,8 ± 258,92 neurônios/8,96 mm2 na região mesentérica. As incidências de neurônios pequenos, médios e grandes foram 14,4, 82 e 3,6% na região antimesentérica, e 14,6, 70,8 e 14,4% na região mesentérica, respectivamente. Concluiu-se que, em ratos Holtzman adultos, predominam o arranjo ganglionado no plexo mioentérico e neurônios NADH-dp de tamanho médio. Os resultados encontrados indicam que os neurônios mioentéricos NADH-dp do íleo de ratos Holtzman são similares aos dos ratos da linhagem Wistar no que diz respeito à localização, arranjo ganglionado e predominância de neurônios com PC de tamanho médi