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Item type: Item , TFinder app: artificial intelligence to diagnose tick fever agents and assess parasitemia/bacteremia in bovine blood smears(2025) Trindade, Artur Siqueira Nunes; Morais, Igor Maciel Lopes de; Leal, Luccas Lourenzzo Lima Lins; Salvador, Vanessa Ferreira; Heller, Luciana Maffini; Beltrán Zapa, Dina María; Aquino, Lídia Mendes de; Ferreira, Lorena Lopes; Lopes, Welber Daniel ZanettiDue to the importance of diagnosing tick fever (TF) agents and their parasitemia in the field to provide appropriate treatment, the objective of this study was to develop an application capable of detecting the presence or absence of these hemopathogens and calculating parasitemia/bacteremia. Therefore, to create the TFinder app, blood smears from the tip of the tail of cattle naturally infected with TF were prepared to train the artificial neural network (ANN) for Anaplasma marginale and Babesia spp. The ANN was trained with images from different microscope fields and angles. For the validation stage of the ANN, new blood smear images from different animals were inserted. The diagnosis of presence or absence and calculation of parasitemia/bacteremia performed by a human being were compared with that of the ANN. The ANN was trained with 8100 and 2871 images of blood smears containing parasitized erythrocytes with A. marginale and Babesia spp. respectively. In the validation stage, it was used a total of new 1000 for A. marginale and 750 for Babesia spp. There was a strong positive correlation between the A. marginale bacteremia (r = 0.9484; R² = 0.8996; p < 0.0001) values obtained by the human and TFinder app, the same occurred for Babesia spp. parasitemia (r = 0.9650; R² = 0.9314; p < 0.0001). The sensitivity, specificity, and accuracy of TFinder app for the presence or absence of A. marginale were 97.7 %, 86.5 %, and 95.4 %, respectively. While for A. marginale bacteremia it was 89.6 %, 98.7 %, and 98.1 %, respectively. For Babesia spp., the sensitivity, specificity, and accuracy of presence and absence were 98.0 %, 91.1 %, and 96.9 %, respectively. The calculation of Babesia spp. parasitemia was 84.3 %, 99.7 % and 98.9 %, respectively. TFinder app can be used to diagnose and calculate the parasitemia/bacteremia of TF agents in bovine blood smear images.Item type: Item , Parasitological, serological and molecular evaluation of Trypanosoma vivax in cattle experimentally infected and early treated with isometamidium chloride in a region lacking the biological vector(2025) Iuasse, Haryie Victória; Salvador, Vanessa Ferreira; Morais, Igor Maciel Lopes de; Leal, Luccas Lourenzzo Lima Lins; Couto, Luiz Fellipe Monteiro; Trindade, Artur Siqueira Nunes; Tamiozo, Gabriel Lopes; Aquino, Lídia Mendes de; Bastos, Thiago Souza Azeredo; Machado, Rosangela Zacarias; Cadioli, Fabiano Antonio; Lopes, Welber Daniel Zanetti; Ferreira, Lorena LopesThis study aimed to evaluate whether early treatment with isometamidium chloride (ISM) affects the production of anti-Trypanosoma vivax antibodies in cattle and whether it can induce parasitological cure, based on tissue analysis 546 days post-treatment in animals without evidence of parasitological relapse. Ten male cattle were experimentally infected with ≈ 1 × 106 viable T. vivax trypomastigotes (D-6) and treated with ISM six days post-infection (D0). The animals were euthanized 546 days post-treatment. During this period, the presence of T. vivax in blood was assessed using direct methods (Woo, Brener, and conventional PCR - cPCR) and serological analysis (indirect iELISA - iELISA). Liver, spleen, brain, heart and pre-scapular lymph node were collected and examined using cPCR. Molecular analysis of blood detected T. vivax DNA up to the day of treatment, but none thereafter. No T. vivax relapses in bloodstream was detected. Anti-T. vivax antibodies were detected from 42 to 546 days post-treatment, except in one animal. However, T. vivax DNA was still detected in the liver, brain, heart, and spleen of at least one animal at 546 days post-treatment. These findings suggest that early ISM treatment does not achieve complete parasitological clearance, as both antibodies and parasite DNA were detected.Item type: Item , Can the tick Rhipicephalus microplus transmit Trypanosoma vivax to cattle by the transovarial and mechanical routes?(2025) Couto, Luiz Fellipe Monteiro; Beltrán Zapa, Dina María; Salvador, Vanessa Ferreira; Morais, Igor Maciel Lopes de; Leal, Luccas Lourenzzo Lima Lins; Trindade, Artur Siqueira Nunes; Bastos, Thiago Souza Azeredo; Tamiozo, Gabriel Lopes; Aquino, Lidia Mendes de; Cardoso, Ennya Rafaella Neves; Lopes, Welber Daniel ZanettiDue to the presence of Trypanosoma vivax DNA in ticks, transovarian and mechanical transmission is suspected. The present study aimed to evaluate (i) the presence of T. vivax in R. microplus that fed on animals with acute trypanosomosis; (ii) the transovarian transmission; and (iii) the ability of larvae and males to mechanically transmit T. vivax to cattle in a region without a cyclic vector. To this end, six calves were experimentally infected with T. vivax on day 0, and after different intervals, these animals were infested with cattle tick larvae. Before oviposition, the content of engorged females was submitted to blood smear to detect trypomastigotes. After oviposition, the engorged females were tested for T. vivax DNA. To assess transovarian transmission, pools of egg masses and larvae from these females were also tested for T. vivax DNA. The larvae were then fed on three negative animals. In addition, to assess mechanical transmission, males from the infestation of animals with trypanosomosis were removed and placed on three other T. vivax-negative calves to feed for 25 days. The male ticks were also tested for T. vivax DNA. The six animals infected with T. vivax showed parasitemia 5-21 days post-infection, when they were treated with a trypanocide drug. Trypanosoma vivax trypomastigotes and DNA were detected in engorged females of R. microplus, but not in egg masses and larvae from engorged females that fed on calves positive for this protozoon. In males, no T. vivax DNA was detected after they fed on cattle positive for T. vivax, and no mechanical transmission was observed.Item type: Item , First study confirming the transmission of Trypanosoma vivax in cattle through rectal palpation: how many animals can be infected by reusing the same procedure glove, and what role does iodine play as a disinfectant?(2025) Leal, Luccas Lourenzzo Lima Lins; Bastos, Thiago Souza Azeredo; Morais, Igor Maciel Lopes de; Heller, Luciana Maffini; Salvador, Vanessa Ferreira; Trindade, Artur Siqueira Nunes; Couto, Luiz Fellipe Monteiro; Beltrán Zapa, Dina María; Bittencourt, Raphaela Bueno Mendes; Ferreira, Lorena Lopes; Lopes, Welber Daniel ZanettiThe aim of this study was to investigate whether Trypanosoma vivax is transmitted via rectal palpation in cattle, using the same glove with different blood scores, from an animal with an acute infection of this protozoan. In addition, the efficacy of iodine as disinfectant, together with water with or without the presence of feces, to prevent the transmission of T. vivax in cattle during the rectal palpation process was evaluated in the laboratory and animals. A total of four stages were carried out, using a total of 103 animals. Stage 1 aimed to verify the presence of trypomastigote forms of T. vivax in palpation gloves after rectal palpation. Stage 2 consisted of checking the blood scores on a palpation glove and how many animals become infected with T. vivax through rectal palpation by reusing the same glove with different blood scores. Stage 3 verified the disinfectant action of 0.5% iodine in the laboratory and in the field. Stage 4 verified the disinfectant action of 1% iodine in the field. Based on the results found, for the first time, the transmission of T. vivax via rectal palpation in cattle was proven using the same glove. From one animal with acute infection by this protozoan, two to five other cattle became infected, depending on the glove blood scores. Disinfecting the glove in a 0.5% iodine solution inhibited the transmission rate of T. vivax among cattle by 70 to 80%. Disinfecting the glove in 1% iodine solution was 100% effective. This work fills gaps in the existing literature, as the transmission of T. vivax via rectal palpation in cattle was demonstrated using the same glove.Item type: Item , First evaluation of the efficacy and operational cost of acaricidal bath combining different droplet sizes, pressures and spray patterns in dairy cows naturally infested with Rhipicephalus microplus(2025) Souza, Gabriel Resende; Araujo, Beatriz Rezende Gandra de; Coelho, Sandra Gesteira; Araújo, Ricardo Nascimento; Lopes, Welber Daniel Zanetti; Rodrigues, Daniel Sobreira; Ferreira, Lorena LopesRhipicephalus microplus poses a significant challenge to animal health. This study evaluated efficacy and the effective operational cost (EOC) of different acaricidal treatments varying droplet size, pressure and spray pattern using a power sprayer. Three experiments were conducted on dairy cows, with each experiment involving three groups of 10 animals: two treated groups and one untreated control group. In the first experiment, acaricide treatments with the same droplet size (medium) were compared at different pressures (50 and 100 psi) using a full cone spray pattern: E1-M50 and E1-M100. In the second experiment, different droplet sizes (fine and medium) were compared at the same pressure (100 psi) using a full cone spray pattern: E2-F100 and E2-M100. The third experiment compared droplet sizes and spray patterns (fine at full cone and ultra coarse at flat fan) at the same pressure (100 psi): E3-F100 and E3-UC100. All animals received acaricide treatment on day 0 with a product based on organophosphate and pyrethroid. Tick counts were performed on days + 7, + 14, and + 21 to determine treatment effectiveness. Data on bath time were collected to determine the EOC, besides cost with acaricide, labor, water and electricity. No statistical differences (p > 0.05) regarding the mean tick count were observed between treated groups in all experiments. However, the highest efficacy values were found in treatments with larger droplets (medium and ultra coarse) and higher pressure (100 psi). In experiment 1, the efficacy varied from 70.6 to 88.9% and 75.2 to 93.7% for the treated groups E1-M50 and E1-M100, respectively. In experiment 2, efficacy varied from 48.7 to 86.2% for E2-F100 and from 68 to 94.7% for E2-M100. In experiment 3, efficacy ranged from 62.9 to 76.9% for E3-F100 and from 73.4 to 86.5% for E3-UC100. The EOC of the acaricide bath varied between 0.42 and 0.46 USD per animal. The findings show that treatment efficacy and EOC vary depending on the spray parameters used for the control of R. microplus, but further studies should be conducted to determine better combinations of pressure, droplet size and spray pattern.Item type: Item , Efficacy of macrocyclic lactones against gastrointestinal nematodes of bovines and their effect on eggs in the uterus of females of these parasites(2025) Beltrán Zapa, Dina María; Maciel, Willian Giquelin; Teixeira, Weslen Fabricio Pires; Lucas, Gustavo Felippelli; Gomes, Vinicius Costa; Ferreira, Lorena Lopes; Amarante, Alessandro Francisco Talamini; Costa, Alvimar José da; Soares, Vando Edesio; Lopes, Welber Daniel Zanetti; Borges, Fernando de AlmeidaThis study evaluated the effect of the injectable macrocyclic lactones (ML) ivermectin (IVE), abamectin (ABA), eprinomectin (EPR), doramectin (DOR) and moxidectin (MOX) on the number of eggs present in the uterus of strongyles from naturally infected cattle. In addition, the efficacy and diagnosis of resistance were assessed. Animals were treated (n = 8 for each ML) on day 0 of the study and necropsied on D+ 14. The number of eggs present in the uterus of the females and the lengths of Haemonchus placei, Oesophagostomum radiatum, Trichostrongylus axei and Cooperia spp. were measured. In addition, fecal egg counts (FEC) were performed, and the results showed efficacies for IVE, ABA, EPR, DOR and MOX, respectively, of 88.9 %, 76.9 %, 58.2 %, 62.3 % and 92.5 %. Through CET, H. placei and C. punctata populations were diagnosed as resistant to all ML tested except MOX. The populations of C. pectinata were resistant to ABA, EPR and DOR, while results were inconclusive for IVE and MOX. T. axei was resistant to IVM, while O. radiatum was resistant to all ML except IVM. In general, all ML significantly (p < 0.05) reduced the mean number of eggs in the uterus of females collected from treated animals (ranging from 11.1 % for IVER/O. radiatum to 94.2 % for MOX/T. axei), except MOX for O. radiatum. The mean length of the females collected from treated animals was shorter (p < 0.05) than in the control group. When the parasite resistance process is incipient for the most common and prolific strongyle species in each population, the clinical diagnosis of resistance through the fecal egg count reduction test (FECRT) showed some inconsistencies compared to the controlled efficacy test (CET).Item type: Item , Tick fever agents in dairy cattle from a tropical region subjected to different breeding systems: parasitological, clinical, serological aspects and genetic diversity of Anaplasma marginale(2025) Cavalcante, Alliny Souza de Assis; Ribeiro, Nathália Braz; Paula, Warley Vieira de Freitas; Sales, Kamila Gaudêncio da Silva; Machado, Rosangela Zacarias; André, Marcos Rogério; Calchi, Ana Cláudia; Krawczak, Felipe da Silva; Ferreira, Lorena Lopes; Bastos, Camila de Valgas e; Lopes, Welber Daniel ZanettiThis study aimed to evaluate the infection dynamics by TF agents in heifers from birth to first calving, in contact with R. microplus, correlating parasitological, serological and clinical aspects of the infection. We also evaluate the persistence of the infection by TF agents in cows raised with or without contact with R. microplus, and the genetic diversity of A. marginale. Three studies were performed. In the first, 15 Girolando heifers (15/16 Holstein × 1/16 Gyr) were used and monitored from birth until their first calving (start of lactation), totaling 28 months of monitoring. In study 2, cows between the 1st and 6th lactation, which were raised on Compost Barn during lactation and on pasture during the dry period, were evaluated. In study 3, cows from the 1st to 7th lactation that after the first calving had no more contact with the cattle tick were evaluated. The animals were evaluated for parasitological, clinical, serological aspects and genetic diversity of Anaplasma marginale. Calves raised on pasture in contact with R. microplus in a tropical region reached a TF agent positivity ≥75 % at 4 months of age. TF cases occurred mostly after weaning, between 3 and 8 months of age. There was a correlation in A. marginale bacteremia between qPCR and blood smear, but no correlation between bacteremia and iELISA. Calves exposed to ticks between 3 and 8 months of age tended not to develop clinical TF as adults, even when continually exposed to R. microplus. Serologic or molecular results for B. bovis and B. bigemina in cows should be interpreted with caution, as the presence of these agents in less than 75 % of the herd may not indicate enzootic instability. In addition, cows exposed to ticks during dry-off had infections with higher levels of A. marginale in the 5th and 6th lactations. On the other hand, cows that had no further contact with R. microplus after their first calving showed infections with higher A. marginale levels in the first lactation. The A. marginale msp4 genotypes reported herein are the ones previously reported in Brazil.Item type: Item , Pharmacokinetics and efficacy of isometamidium chloride against Trypanosoma vivax in cattle(2025) Couto, Luiz Fellipe Monteiro; Beltrán Zapa, Dina María; Heller, Luciana Maffini; Bastos, Thiago Souza Azeredo; Vettorato, Luis Fernando; Amaral, Heitor de Oliveira Arriero; Chiummo, Rafael Marin; Rodrigues, Daniel de Castro; Soares, Vando Edesio; Ferreira, Lorena Lopes; Lopes, Welber Daniel ZanettiWe evaluated the pharmacokinetics (pk), preventive efficacy (PE) and therapeutic efficacy (TE) of isometamidium chloride (ISM) against T. vivax. For the pk study, the animals received ISM 1 mg/kg (1 mL/40 kg) on day 0 and plasma samples were collected at day 0 (immediately before treatment), 1 h, 3 h 6 h, 9 h and 12 h after treatment, D + 1 (24 h and 36 h), D + 2 (48 h), D + 7, D + 14, D + 21, D + 28, D + 42, D + 56 and D + 68 post treatment. Plasma samples were analyzed using mass spectrometry coupled to ultra-performance liquid chromatography (UPLC). As result for pk study, the ISM 1 mg/kg reached the highest plasma concentrations 1 h after administration (488.57 ± 208.59 μg/L) and was found in plasma until 24 h. No further detection of ISM in plasma happened beyond 36 h after treatment. For the PE study, twenty-four bovines divided in four groups of six animals each. All groups received ISM 1 mg/kg (1 mL/40 kg) on different days. The T01 group received ISM on day -120, while the T02 animals were treated with ISM on day -90, T03 was treated with ISM on day -60, and the control group (T04) consisted of two animals that received saline solution on day -120, two other animals that received the solution on day -90, and the last two animals that received saline on day -60. In the PE study, the experimental infection with 1 × 106 trypomastigotes of T. vivax of the 24 animals occurred on day 0. Until day 30 after infection, the analysis of T. vivax was performed by the Woo, Brener and cPCR methods, to determine the PE of ISM after 60, 90 and 120 days. As result for the PE study, the PE of ISM against T. vivax was 100 % up to 90 days. For the TE study, 16 animals were experimentally infected with T. vivax trypomastigotes on day -5. On Day 0, the animals were divided into two groups or eight animals each: T01 treated with ISM 0.5 mg/kg (1 mL/80 kg), and T02 treated with saline solution (control). Blood samples after treatment were again analyzed by the Woo, Brener and cPCR methods to determine TE of ISM. Furthermore, in the TE study, to prove that the animals in the treated group were not infected with T. vivax, a biological test was performed on young goats on day +45. As result for the TE study, the TE of ISM 0.5 mg/kg was 100 %, and no young goats were infected. In conclusion, the commercial ISM-based product tested here can be used both preventively and therapeutically against T. vivax. We evaluated the pharmacokinetics (pk), preventive efficacy (PE) and therapeutic efficacy (TE) of isometamidium chloride (ISM) against T. vivax. For the pk study, the animals received ISM 1 mg/kg (1 mL/40 kg) on day 0 and plasma samples were collected at day 0 (immediately before treatment), 1 h, 3 h 6 h, 9 h and 12 h after treatment, D + 1 (24 h and 36 h), D + 2 (48 h), D + 7, D + 14, D + 21, D + 28, D + 42, D + 56 and D + 68 post treatment. Plasma samples were analyzed using mass spectrometry coupled to ultra-performance liquid chromatography (UPLC). As result for pk study, the ISM 1 mg/kg reached the highest plasma concentrations 1 h after administration (488.57 ± 208.59 μg/L) and was found in plasma until 24 h. No further detection of ISM in plasma happened beyond 36 h after treatment. For the PE study, twenty-four bovines divided in four groups of six animals each. All groups received ISM 1 mg/kg (1 mL/40 kg) on different days. The T01 group received ISM on day -120, while the T02 animals were treated with ISM on day -90, T03 was treated with ISM on day -60, and the control group (T04) consisted of two animals that received saline solution on day -120, two other animals that received the solution on day -90, and the last two animals that received saline on day -60. In the PE study, the experimental infection with 1 × 106 trypomastigotes of T. vivax of the 24 animals occurred on day 0. Until day 30 after infection, the analysis of T. vivax was performed by the Woo, Brener and cPCR methods, to determine the PE of ISM after 60, 90 and 120 days. As result for the PE study, the PE of ISM against T. vivax was 100 % up to 90 days. For the TE study, 16 animals were experimentally infected with T. vivax trypomastigotes on day -5. On Day 0, the animals were divided into two groups or eight animals each: T01 treated with ISM 0.5 mg/kg (1 mL/80 kg), and T02 treated with saline solution (control). Blood samples after treatment were again analyzed by the Woo, Brener and cPCR methods to determine TE of ISM. Furthermore, in the TE study, to prove that the animals in the treated group were not infected with T. vivax, a biological test was performed on young goats on day +45. As result for the TE study, the TE of ISM 0.5 mg/kg was 100 %, and no young goats were infected. In conclusion, the commercial ISM-based product tested here can be used both preventively and therapeutically against T. vivax.Item type: Item , Ecotoxicity of microplastics in embryos and newly hatched snails of Biomphalaria glabrata(2025) Harayashiki, Cyntia Ayumi Yokota; Rodrigues, Cândido Carvalho; Rocha, Thiago LopesMicroplastics (MPs) are emerging pollutants widely found in aquatic ecosystems. Despite the increasing number of studies concerning the effects of MPs on aquatic organisms, few have focused on toxicity in the early developmental stages of gastropods living in freshwater ecosystems. Thus, the current study aimed to evaluate the effects of polyethylene MPs (PE-MPs) in the neotropical gastropod Biomphalaria glabrata embryos and newly hatched snails (NH). The ecotoxicity of PE-MPs (from 0.3 mg L− 1 – environmental relevant concentration – to 160 mg L− 1) was analyzed by Biomphalaria embryotoxicity test (BET) (192 h) and acute toxicity tests with NH (96 h). Mortality and behavior (locomotor and touch sensitivity) were assessed in both embryos and NH, while heart rate, hatching rate, embryo development, reactive oxygen species – ROS levels, and cell death were analyzed during BET. Results showed that PE-MPs did not induce mortality or changes in the hatching rate, developmental stages, and behavior of B. glabrata. However, sub-lethal effects were observed regarding heart rate, ROS, and cell death of embryos. These results indicate that embryos are more sensitive than NH. In addition, the lack of effects on hatched snails from BET suggests that the effects observed during their development were not strong enough to affect the completely formed snail, which could be related to additional protection provided by the gelatinous substance that composes the egg clutches.Item type: Item , Genotoxicity, mutagenicity and immunotoxicity assessment of microplastics and nanoparticle mixture in bullfrog tadpoles(2025) Jacintho, Jaqueline Cardoso; Pedroso, Thays Millena Alves; Lopes, Patrick Cardoso; Rocha, Thiago Lopes; Jacob, Raquel Fernanda SallaAmphibians are among the most endangered vertebrates whose population declines are linked to environmental pollution, including emerging pollutants such as microplastics (MPs) and nanoparticles (NPs). However, knowledge about the interactive effects of MPs and titanium dioxide (TiO2) NPs on amphibians remain scarce. This study aimed to evaluate the potential genotoxicity, mutagenicity and immunotoxicity of polyethylene (PE) MPs (60 mg L−1) and TiO2 NPs (10 μg L−1), individually or in a mixture (Mix) in Aquarana catesbeiana tadpoles, Gosner stage 25, after a sub-chronic exposure (15 days). Multiple parameters were assessed by the comet assay, micronucleus test and nuclear abnormalities in erythrocytes, and differential leukocyte count. The individual pollutants and the Mix did not cause genotoxicity. The total number of nuclear abnormalities, the frequency of notched nucleus and moved nucleus were higher in individually PE MPs and TiO2 NPs groups, but not in Mix. Both pollutants, individually and in a mixture, did not alter the leukocyte profile of the tadpoles. Overall, our results reinforce the complexity of the outcomes of emerging pollutant mixtures, which represent a new environmental issue to discuss about amphibians' health and ecotoxicology.Item type: Item , Differential biochemical responses of Cyprinus carpio after dietary and waterborne exposure to microplastics from polyethylene-based biodegradable and conventional bags(2025) Storck, Tamiris Rosso; Ames, Jaíne; Qualhato, Gabriel; Vasconcelos, Marlon de Castro; Piccoli, Andreli Lopes; Rocha, Thiago Lopes; Fernandes, Andréia Neves; Clasen, Barbara EstevãoMicroplastics (MPs) can interact and generate toxic effects on exposed organisms, such as fish. However, little is known about the effects of exposing fish to biodegradable PE-MPs, and how the form of exposure is related to toxicity. The current study aimed to evaluate the impact of dietary and waterborne exposure to four types of polyethylene MPs (PE-MPs) from biodegradable and conventional bags on common carp Cyprinus carpio. The toxicity of PE-MPs from biodegradable (PE-BIO), transparent (HDPE-1), black (PE-OXO), and gray (HDPE-2) bags was analyzed after 7 days of exposure by multiple biochemical biomarkers assessment, such as neurotoxicity, oxidative and nitrosative stress, and oxidative damage. The toxicity of waterborne exposure of fish to PE-MPs followed the order: PE-OXO>HDPE-2>PE-BIO>HDPE-1, while the dietary exposure was: HDPE-2>PE-BIO>HDPE-1>PE-OXO. Furthermore, according to IBRv2, the HDPE-2 and PE-BIO treatments of dietary exposure showed greater changes in biomarkers compared to the control. Lipid peroxidation levels were altered only in dietary exposure treatments, while CAT and GST activity was generally inhibited in both exposure pathways. The PE-BIO treatments showed significant differences in relation to conventional plastics, mainly in the nitric oxide (NO) content of the fish liver, which increased or decreased depending on the route of exposure. Therefore, this pioneering study emphasizes that the composition of PE-MPs and the route of exposure can generate different effects on C. carpio biomarkers. Furthermore, these results may indicate the effects of environmental contamination by MPs in a freshwater fish species.Item type: Item , Developmental toxicity of formulated insecticide mixture containing imidacloprid and beta-cyfluthrin in fish: insights using zebrafish(2025) Figueirêdo, Livia Pitombeira de; Cirqueira, Felipe; Sousa, Bianca Leite Carnib de; Mamboungou Junior, Joseph; Rocha, Thiago LopesInsecticides are critical in controlling pests and disease vectors. However, there is still a lack of ecotoxicological studies using commercial formulations of insecticides containing active ingredients. The study aimed to evaluate the developmental toxicity of a commercial insecticide mixture (imidacloprid [IMI] + beta-cyfluthrin [β-CYF]). Mortality, hatching rate, spontaneous contraction, heartbeat, morphological changes, reactive oxygen species (ROS), skeletal development, and locomotor behavior of zebrafish were analyzed. Embryos were exposed to imidacloprid (IMI) and β-cyfluthrin (β-CYF) in the following ratios: 0.001 mg IMI·L−1 + 0.000125 mg β-CYF·L−1 (C1); 0.01 mg IMI·L−1 + 0.00125 mg β-CYF·L−1 (C2); 0.1 mg IMI·L−1 + 0.0125 mg β-CYF·L−1 (C3); 1.0 mg IMI·L−1 + 0.125 mg β-CYF·L−1 (C4); 10.0 mg IMI·L−1 + 1.25 mg β-CYF·L−1 (C5) for 144 h. The results showed a mortality of 50 % of organisms in the C5 concentration. Embryos exposed to C1 and C3 showed tachycardia and hatched early compared to the negative control, indicating cardiotoxic and embryotoxic effects. The two highest concentrations tested (C4 and C5) induced evident morphological changes (yolk sac and pericardial edema, and spine alterations), and skeletal toxicity (absence of cartilage and bone formation), along with decreased larval swimming behavior. Also, the formulated insecticide (C1) increased ROS levels in zebrafish larvae. Results showed that the formulated insecticide containing IMI and β-CYF induces several toxic effects on developing zebrafish, indicating its environmental risk to aquatic organisms.Item type: Item , Genotoxicity, mutagenicity, and immunotoxicity assessment of green silver nanoparticles from Croton urucurana on neotropical snails Biomphalaria glabrata(2025) Araújo, Paula Sampaio; Caixeta, Maxwell Batista; Qualhato, Gabriel; Gonçalves, Bruno Bastos; Gomes, Rodrigo Saar; Viali, Eloiza da Silva Nune; Viali, Wesley Renato; Rocha, Thiago LopesGreen silver nanoparticles (G-Ag NPs) have been indicated as safer nanotechnology in nanomedicine, especially for controlling microorganisms, vectors, and gastropods of medical importance. However, their mechanisms of action and ecotoxicity in aquatic organisms remain unknown. Thus, the current study aimed to investigate the effects of G-Ag NPs synthesized through Croton urucurana leave aqueous extract on the neotropical freshwater snail Biomphalaria glabrata after in vivo exposure using multi-biomarker responses in hemocytes. Adult snails were exposed to G-Ag NPs (0.05 and 0.14 mg L−1) or leave aqueous extract (5.4 mg L−1) for 7 days. Genotoxicity (comet assay), mutagenicity (micronucleus test and nuclear abnormality assay), and immunotoxicity (phagocytosis assay by flow cytometry) were analyzed in hemocytes. Results showed that G-Ag NPs induced high mortality and genotoxic effects (DNA damage) compared to leave aqueous extract and negative control groups. G-Ag NPs also induced mutagenic effects (nuclear alterations), mainly notched and blebbed nuclei. Also, G-Ag NPs and extract did not change the phagocytic activity and cell viability of hemocytes. These findings contribute to understanding the toxicity of green nanoparticles in freshwater gastropods and represent a pioneering effort to evaluate the effects of G-Ag NPs and the C. urucurana aqueous extract in B. glabrata hemocytes. Overall, the immune system of gastropods is a target of green nanoparticle toxicity.Item type: Item , Glucocorticoids as emerging pollutants in surface water: a systematic review on their global occurrence and distribution(2025) Silva, Marco Antonio Lima e; Silva, Rhitor Lorca da; Teixeira, Tiago Porfírio; Rocha, Thiago Lopes; Marcon, Matheus FelipeGlucocorticoids (GCs) are synthetic drugs widely used for treating several diseases with inflammatory pathophysiology. In general, 1–20% of the concentration of GCs initially administered is eliminated unchanged in the urine while still in its active form, and therefore, fractions of GCs are constantly released in effluents. Currently, water treatment plants do not have sufficiently effective technologies to remove these substances completely, favoring the presence of these emerging pollutants in the effluents of wastewater treatment plants. In this context, we conduct a systematic review to identify studies that found GCs in surface water. The general data of the included studies, the GCs found and their concentration, the water body where the GCs were found, and the place and date of sampling were summarized. GCs have already been found in the surface water of 24 countries, mainly China, Malaysia, and the United States. Countries with the highest concentrations of GCs found in surface water were Mexico, India, and Brazil. Betamethasone, budesonide, corticosterone, cortisol, cortisone, dexamethasone, fludrocortisone, fluticasone, hydrocortisone, methylprednisolone, prednisolone, prednisone, and triamcinolone were reported at concentrations ranging from 0.00098 to 24760 ng/L, including in water for human consumption. Revised data showed that the real presence of these substances worldwide is still underestimated, requiring further studies to determine their real distribution. Furthermore, we believe that some strategies can be adopted to mitigate surface water contamination by GCs, such as the intensification of public programs about educational activities related to the rational use of medicines, and the urgent need to improve and expand the water treatment methods.Item type: Item , Ecotoxicological impact of residual polycyclic aromatic hydrocarbon pollution in the largest Brazilian marine protected area: a multibiomarker approach in the marine clam Tivela mactroides(2025) Sousa, Bianca Leite Carnib de; Oliveira, Jerusa Maria de; Nascimento, Zaíne Marcyelle Freire do; Garibaldi, Gabriel Tenorio Toledo; Pinto, Taciana Kramer de Oliveira; Rocha, Thiago Lopes; Cavalcante, Rivelino Martins; Lemos, Viviane Agostinho da Silva; Bastos, Luciana Machado; Nossol, Arlene Bispo dos SantosAn unprecedented oil spill occurred along the Northeast Coast of Brazil in late 2019, affecting the Environmental Protection Area Costa dos Corais (APACC), the largest Brazilian marine conservation unit, posing a risk to this complex ecosystem. By applying passive ecotoxicological biomonitoring, this study aimed to evaluate whether the remaining contamination from such an oil spill affected the population of the marine clam Tivela mactroides by assessing multiple biomarkers, regarding antioxidant enzyme activities (SOD, CAT), biotransformation (GST, GSH), oxidative damage (MDA), and neurotoxicity (AChE) in gills and digestive glands, and PAH bioaccumulation. The clam tissues and sediment were sampled in three locations (Paripueira, Japaratinga and Maragogi beaches), which were differently affected by the accident, and during three sampling periods, starting one year after the accident began. We detected Σ16PAHs in the clam soft tissues and in the sediment samples, all with relatively low concentrations. Differential biomarker responses were observed in both clam tissues, as denoted by the Integrated Biomarker Response Index (IBR). However, there was no correlation between PAH bioaccumulation and biomarker responses in T. mactroides, suggesting the occurrence of multiple stress factors in such a marine protected area. This study demonstrated that multiple biomarkers assessment in the marine clam T. mactroides is a suitable approach for biomonitoring of marine areas in Brazil affected by the 2019 oil spill. However, we emphasize that further long-term studies must be conducted to better understand the environmental impacts caused by an oil spill of such magnitude.Item type: Item , MICROMar Project: the largest standardized assessment of microplastic pollution across southwestern Atlantic coastal ecosystems(2025) Giarrizzo, Tommaso; Santos, Robson Guimarães dos; Reis Filho, José Amorim dos; Martinelli Filho, José Eduardo; Barbosa, Neuciane Dias; Nunes, Jorge Luiz Silva; Rocha, Thiago Lopes; Jacobucci, Giuliano Buzá; Rodrigues, Aline Sueli de Lima; Abraão, Letícia Paiva de Matos; Guimarães, Tiago Batista; Fonseca, Estefan Monteiro daItem type: Item , Metazoan endoparasites of Myloplus nigrolineatus (Characiformes: Serrasalmidae) from upper Amazon river basin, Brazil(2025) Amaral, Rafael Braga do; Araujo, Guilherme Augustus de; Silva, Luciana Damacena; Rocha, Thiago Lopes; Takemoto, Ricardo Massato; Marco Júnior, Paulo DeThe success of Trematoda and Nematoda infection in fish involves a complexity of variables. The objective of this study was to report the parasitological descriptors (prevalence, mean abundance, and mean intensity) and community status of Myloplus nigrolineatus as well as to evaluate the relationship between abundance and richness of endoparasites with biometric parameters, sex, Kn the hosts, percentage cover native vegetation and water temperature. A total of 7,256 endoparasites were found: 861 digeneas and 6,395 specimens of nematodes were collected in the intestine from Myloplus nigrolineatus. One species of Digenea and five species of Nematoda were collected. The initial documentation of the prevalence (%) of endohelminth species in M. nigrolineatus revealed the following: Dadaytrema oxicephala (70%), Chabaudinema americanum (52%), Cucullanus pinnai pinnai (13%), Myleusnema bicornis (65%), Procamallanus (Spirocamallanus) inopinatus (9%) and Rondonia rondoni (35%). The mean intensity and mean abundance were 329.82 ± 416 and 315.48 ± 417, respectively. The trematode Dadaytrema oxycephala was considered a central and dominant species. The total abundance was explained by the variables total length, relative condition factor (Kn), percentage cover native vegetation and water temperature. Fish relative condition factor (Kn) and sex were not influenced by the parasite infection and did not impair the body condition of the hosts. The GLMM showed there is no relationship between abundance and richness of endoparasites with percentage cover native vegetation, while that every twenty centimeters more in the total length of the hosts, the abundance of endoparasites in the intestine increases, approximately, 2 specimens.Item type: Item , Single and combined chronic toxicity of cadmium and titanium dioxide nanoparticles in Aquarana catesbeiana (Anura: Ranidae) tadpoles(2025) Moraes, Carolina Arantes de; Oliveira, Fagner das Neves de; Qualhato, Gabriel; Brito, Pedro Vale de Azevedo; Simões, Karina; Rocha, Thiago Lopes; Jacob, Raquel Fernanda Salla; Vieira, Lucélia GonçalvesThe rising production and improper disposal of titanium dioxide nanoparticles (TiO2 NPs) into aquatic systems present considerable environmental challenges, especially when these particles interact with other contaminants such as cadmium (Cd). Thus, the current study aimed to evaluate the potential toxic effects on the gills, chondrocranium, body growth, and mortality of Aquarana catesbeiana tadpoles. The tadpoles were exposed to environmentally relevant concentrations of TiO2 NPs (10 µg L-1), and CdCl2 (10 µg L-1), both individually and in combination, for 30 days (chronic exposure), along with a control group. Our results indicate that the co-exposure to TiO2 NPs and Cd induced a higher mortality rate. In the gills, TiO2 NPs led to epithelial simplification, while Cd exposure resulted in stratified epithelium formation. Additionally, there were notable changes in the index of degenerative alterations for the co-exposed group and the overall organ index for the groups exposed to Cd and the mixture. The viscerocranium showed significant malformations in the ceratobranchials and reticular processes, indicating the mixture's toxicological potential during the skeletal system's development. Morphometric analysis also revealed reduced body length and abnormal body ratios in tadpoles from the co-exposure group. In conclusion, TiO2 NPs and Cd, both alone and in combination, exhibit toxicological effects in A. catesbeiana tadpoles, indicating a potential ecological risk associated with releasing these contaminants into aquatic environments.Item type: Item , A systematic review of estrogens as emerging contaminants in water: a global overview study from the one health perspective(2025) Silva, Rhitor Lorca da; Silva, Marco Antonio Lima e; Teixeira, Tiago Porfirio; Assunção, Thaís Soares Farnesi de; Teixeira, Paula Pinheiro; Tamagno, Wagner Antonio; Rocha, Thiago Lopes; Souza, Julio Cesar de; Gonçalves, Inácio; Marcon, Matheus FelipeThe widespread presence of estrogens in aquatic environments represents a One Health concern, as it simultaneously threatens environmental integrity, wildlife health, and human well-being. These compounds, widely used in human and veterinary medicine, are excreted in partially or unmetabolized forms and persist in the environment due to the inefficiency of conventional water treatment systems in removing them. This systematic review provides a global overview of the occurrence of estrogens in water resources. We synthesized data on study characteristics, estrogen compounds detected, their concentrations, types of water bodies, and geographic locations. In total, 39 estrogens, including natural, synthetic, and metabolite forms, were reported at concentrations ranging from 0.002 to 10,380,000.0 ng/L across 40 water body types in 59 countries on all continents. The most frequently detected compounds were estrone, estradiol, and ethinylestradiol. Estrogens were predominantly identified in wastewater treatment plant effluents, rivers, lakes, surface waters, and even drinking water sources. These findings underscore the estrogen contamination and its potential to disrupt endocrine functions across species, posing serious implications for ecosystems. Within the One Health framework, this review highlights the urgent need for integrated strategies to improve water quality monitoring, develop advanced treatment technologies, and update regulatory standards to address the multifaceted risks posed by estrogenic contaminants.Item type: Item , Europium and calcium-co-doped TiO2 nanocrystals: tuning the biocompatibility and luminescence traceability of Drosophila melanogaster(2025) Oliveira, Jerusa Maria de; Almeida, Larissa Iolanda Moreira de; Santos, Francisco Rubens Alves dos; Carvalho, João Paulo Santos de; Barbosa, Amanda Ingrid dos Santos; Costa, Marcus Andrei Rocha Fernandes da; Maciel, Vanessa Tomaz; Souza, Gabriela Leite de; Magalhães, Alysson Nogueira de; Vermelho, Marcos Vinicius Dias; Rocha, Thiago LopesThe incorporation of europium (Eu) ions improves the biocompatibility of TiO2 nanocrystals (NCs) and allows tracking by red luminescence. Calcium doping improves cellular compatibility while also facilitating better interaction with biological systems. Thus, in this work, were synthesized Eu and Ca co-doped TiO2 NCs and physical–chemical and biological properties were investigated. The physical–chemical properties were performed in order to analised the effects of the doping on the crystalline phase of TiO2 morphology, sized, zeta potential, hydrodynamic diameter, and photocatalytic properties. Biological assessments were conducted using in vitro assays with human osteosarcoma cells (SAOS-2) through cytotoxicity assays and in vivo assays with Drosophila melanogaster, where we evaluated the mortality rate during postembryonic development and the luminescence of nanomaterials in vivo. The physical–chemical properties confirmed with success the integration of Ca ions into the TiO2:Eu crystal (TiO2:Eu:xCa) NCs without additional phases. The co-doping of Ca led to a reduction of approximately 70% in photocatalytic activity. Moreover, co-doping with Ca was not cytotoxic to SAOS-2 cells. Our in vivo analysis showed no delays in postembryonic development and no larval or pupal lethality. The larval mortality rate and pupal formation rate were comparable to the control group when D. melanogaster was exposed to nanomaterials at concentrations of 1 mg mL−1 or lower. Luminescence of the NCs was detected in confocal microscopy images, indicating the presence of NCs in the larval brain and intestines. This luminescence was observed in TiO2:Eu:xCa NCs. These results showed that Ca doping improved the biocompatibility and enhanced the luminescence of these materials, making them traceable in biological tissues. Therefore, our research provides valuable insights into the tailored properties of TiO2 for potential applications in various fields of biomedicine.