Resumen En los estados de Michoacán, México y California, EUA, la producción de zarzamora (Rubus sp.) se ve afectada por una marchitez que ocasiona la pérdida de miles de hectáreas. En el presente trabajo se identificó el agente causal de la enfermedad y propuso alternativas de control con microorganismos antagonistas. En el año 2022 de un huerto en Tocumbo, Michoacán, se aisló e identificó morfológicamente al agente causal de la enfermedad. Se realizó un bioensayo preliminar en laboratorio con cepas de B. subtilis, B. amyloliquefaciens, P. fluorescens y Trichoderma spp, nativas de Tacámbaro y Taretan, Mich. Las cepas sobresalientes se probaron en invernadero en plantas de zarzamora cv Tupy de forma preventiva y como control, bajo un diseño experimental de bloques al azar con seis repeticiones. En las pruebas preliminares todas las cepas utilizadas mostraron una inhibición del crecimiento micelial del patógeno superior al 50%, sobresaliendo Trichoderma sp. de Tacámbaro, B. subtilis y P. fluorescens. En las pruebas realizadas in situ los tratamientos aplicados de forma preventiva disminuyeron la incidencia de marchitez desde 57 hasta 66%. En términos de severidad y altura de la planta destacó Trichoderma sp., mientras que el porcentaje de necrosis radicular disminuyó desde 56 hasta 70% con todos los tratamientos. Los tratamientos de control también mostraron eficacia antagonista, aunque en menor medida, disminuyeron la incidencia de marchitez desde 2 al 58% y los porcentajes de necrosis radicular desde 19 a 62%. Los microorganismos antagonistas pueden incorporarse como una estrategia de manejo integral con gran eficiencia.

Abstract In the states of Michoacán, Mexico and California, USA, blackberry (Rubus sp.) production is affected by wilt, which causes the loss of thousands of hectares. In the present work, the causative agent of the disease was identified and control alternatives with antagonistic microorganisms were proposed. In 2022, the causative agent of the disease was isolated and morphologically identified from an orchard in Tocumbo, Michoacán. A preliminary laboratory bioassay was performed with strains of B. subtilis, B. amyloliquefaciens, P. fluorescens, and Trichoderma spp., native to Tacámbaro and Taretan, Mich. The outstanding strains were tested in a greenhouse on plants of blackberry cv Tupy as a preventive measure and as a control, under an experimental design of randomized blocks with six repetitions. In the preliminary tests, all the strains used showed an inhibition of the mycelial growth of the pathogen greater than 50%, with Trichoderma sp. from Tacámbaro, B. subtilis and P. fluorescens standing out. In the tests carried out in situ, preventively applied treatments decreased the incidence of wilt by 57% to 66%. In terms of severity and plant height, Trichoderma sp. stood out, while the percentage of root necrosis decreased by 56% to 70% with all treatments. Control treatments also showed antagonistic efficacy, although to a lesser extent, they decreased the incidence of wilt by 2 to 58% and the percentages of root necrosis by 19 to 62%. Antagonistic microorganisms can be incorporated as a comprehensive management strategy with high efficiency.

Abstract Introduction: Short-term gametes storage is an inexpensive and simple technique that allows the use of the same batch of eggs or sperm at different times, maximizing the application of research protocols and the use of gametes in production. Arbacia dufresnii is a sea urchin species with proven aquaculture potential and already used in the nutraceutical industry. Aging of its gametes is unknown and is a needed information to scale up the production. Objective: Determine the effect of male and female gamete aging on the fertilization success of Arbacia dufresnii. This will allow optimizing the use of gametes after collection decoupling spawning from fertilization. Methods: A. dufresnii individuals were induced to spawn and gametes were kept at 12 ± 1 °C throughout each bioassay. Sperm was separated into two treatments: activated sperm in seawater (AS), and dry sperm (DS). Two bioassays were made: Bioassay 1 evaluated the effect of time on fertility by performing fertilization tests at 0 h, 24 h, 48 h, 72 h, and 96 h after spawning. Bioassay 2 evaluated the contribution of each type of aged gamete on fertility, combining aged gametes (96 h) with fresh gametes (0 h). Results: Bioassay 1: the fertilization success obtained by combining eggs (E) with AS or DS presented important differences. While the fertilization success remained acceptable (greater than 50 %) for up to 72 h using ExDS, it only remained acceptable for up to 48 h using ExAS. Bioassay 2: acceptable fertilization success was found by combining aged E (96 h) with fresh sperm, or aged DS (96 h) with fresh E, but not using aged AS with fresh E. Conclusions: The findings of this work show that fertilization success in A. dufresnii gametes remains relatively unchanged for up to 48 h after spawning when combining ExAS, and for up to 72 h when combining ExDS. However, when combining aged E or aged DS with a fresh gamete, post-collection fertilization can be extended up to 96 h. In this work, the first steps have been taken to understand the conservation time of A. dufresnii gametes with minimum intervention.

Resumen Introducción: El almacenamiento de gametos a corto plazo es una técnica económica y sencilla que permite utilizar el mismo lote de óvulos o espermatozoides en diferentes momentos, maximizando la aplicación de protocolos de investigación y el uso de gametos en la producción. Arbacia dufresnii es una especie con probado potencial acuícola como fuente de gametos para la industria nutracéutica. Sin embargo, se desconoce el envejecimiento de sus gametos y es una información necesaria para escalar la producción. Objetivo: Determinar el efecto del envejecimiento de los gametos masculinos y femeninos en el éxito de la fecundación de Arbacia dufresnii con el fin de optimizar el aprovechamiento de los gametos después de la recolecta desincronizando el desove de la fecundación. Métodos: Se indujo el desove de individuos de A. dufresnii y los gametos se mantuvieron a 12 ± 1 °C durante cada bioensayo. El esperma se separó en dos tratamientos: esperma activado en agua de mar (AS) y esperma seco (DS). Se realizaron dos bioensayos: El Bioensayo 1 evaluó el efecto del tiempo sobre la fertilidad realizando pruebas de fecundación a las 0 h, 24 h, 48 h, 72 h y 96 h después del desove. El bioensayo 2 evaluó la contribución de cada tipo de gameta envejecida (96 h) sobre la fertilidad, combinando gametos envejecidas (96 h) con gametos frescas (0 h). Resultados: Bioensayo 1: el éxito de fecundación obtenido combinando huevos (E) con AS o DS presentó diferencias importantes. Si bien el éxito de la fecundación se mantuvo aceptable (más del 50 %) durante un máximo de 72 h con ExDS, solo permaneció aceptable hasta 48 h con ExAS. Bioensayo 2: se encontró un éxito de fecundación aceptable combinando E envejecidos (96 h) con esperma fresco, o DS envejecido (96 h) con E fresco (0 h), pero no usando AS envejecido con E fresco (0 h). Conclusiones: Los hallazgos de este trabajo muestran que el éxito de la fecundación en los gametos de A. dufresnii permanece relativamente sin cambios hasta 48 h después del desove cuando se combina ExAS, y hasta 72 h cuando se combina ExDS. Sin embargo, cuando se combina E envejecido o DS envejecido con un gameto fresco, el tiempo entre la recolección y la fecundación puede extenderse hasta 96 h. En este trabajo se han dado los primeros pasos para entender el tiempo de conservación de los gametos de A. dufresnii con mínima intervención.

Resumen Este estudio tuvo como objetivo caracterizar las propiedades agroquímicas del orujo de uva (GP) de V. labrusca y evaluar su efecto sobre Azospirillum brasiliense y cultivos hortícolas para determinar su uso seguro como fertilizante y/o mulching. Los bioensayos se realizaron con el GP a diferentes concentraciones para evaluar su toxicidad sobre A. brasiliense y semillas de tomate y lechuga. Para evaluar el efecto de GP sobre el crecimiento de plantas de tomate y lechuga se aplicaron diferentes dosis como mulching en el rango de 20-80 t ha-1. La caracterización agroquímica mostró que GP es rico en potasio y fósforo; y tiene bajo contenido de sales de Na+ (SAR < 15). El GP al 2,5% (p v-1) fue responsable de un aumento significativo en la tasa de supervivencia de las rizobacterias fijadoras de N2. Los resultados de la germinación de las semillas revelaron que la lechuga fue más susceptible al aumento de las concentraciones de GP. La aplicación de 20 t ha-1 de GP en los experimentos de invernadero incrementó la biomasa radicular de lechuga y tomate. Además, la parte aérea del tomate no presentó síntomas de toxicidad. Estos resultados abren la posibilidad de considerar el aprovechamiento de V. labrusca GP como mulching, sin utilizar ninguna tecnología de tratamiento antes de su aplicación al suelo en cultivos de tomate.

Abstract This study characterized the agrochemical properties of V. labrusca grape pomace (GP) and evaluated the effect on the rhizobacteria Azospirillum brasiliense and horticultural crops, determining safety as fertilizer and/or mulching. Two first bioassays were performed with the GP at different concentrations evaluating toxicity on A. brasiliense, and on tomato and lettuce seeds. A third bioassay evaluated GP mulching effects on tomato and lettuce plants growing with amounts varying between 20 and 80 t ha-1. Agrochemical characterization showed that GP is rich in potassium and phosphorus, with a low content of Na+ salts (SAR < 15). The GP at 2.5% (w v-1) significantly increased survival rates of N2-fixing rhizobacteria. Results on seed germination revealed lettuce was more susceptible to increasing GP concentrations. The application of 20 t ha-1 of GP in greenhouse experiments increased lettuce and tomato root biomass. Furthermore, the aerial part of tomato showed no toxicity symptoms. These results open the possibility of considering V. labrusca GP as mulching without prior treatment in tomato crops.

Resumo O objetivo deste trabalho foi avaliar a qualidade do ar com base no fluxo veicular das cidades localizadas em diferentes altitudes na microrregião da Bodoquena, no estado de Mato Grosso do Sul, Brasil. Para tal, foi realizado o teste de micronúcleo, por meio do bioensaio TRAD-MCN em botões jovens de Tradescantia coletadas no período entre fevereiro a novembro de 2018 em sete cidades da microrregião da Bodoquena, com diferentes intensidades de fluxo veicular. Foram avaliados os parâmetros meteorológicos, os veículos foram contados para determinar o tráfego de veículos em cada cidade e altitude. A partir da topografia Shuttle Radar (SRTM) e processamento no software Esri ArcGIS® versão 10.5.1 foi possível mapear a área com base no Modelo de Elevação. As análises morfoanatômicos foram realizadas conforme metodologia padrão. As mensurações de espessura, comprimento, largura dos tecidos e estruturas como a cutícula superior, cutícula inferior, face superior e face inferior da epiderme, hipoderme e mesófilo foram avaliadas. O maior fluxo veicular foi encontrado nas cidades de Bodoquena, Guia Lopes da Laguna, Jardim e Porto Murtinho. O horário das 17:00 às 18:00h foi o que apresentou maiores fluxo de veículos. A maior frequência de alterações mutagênicas foi encontrada na cidade de Guia Lopes, não diferindo de Bonito, Caracol e Jardim. Ao longo do biomonitoramento observou-se que as estações de verão e outono foram as que apresentaram maiores frequências de micronúcleo independente da cidade avaliada. Observou-se que a correlação entre a frequência de micronúcleos e o fluxo veicular foi baixa, apoiando a tese de que essa microrregião, embora apresente alterações mutagênicas em T. pallida, as alterações numericamente são pequenas quando comparadas a outras regiões do estado de Mato Grosso do Sul. Observou-se uma variação na espessura dos tecidos/estruturas que é variável entre as diferentes cidades e estações do ano. De forma geral os tecidos/estrutura apresentaram redução na espessura para as cidades de Nioaque e Porto Murtinho quanto aos aspectos anatômicos e morfológicos, sendo que, para o comprimento e espessura foliar não foi observado diferenças entre as cidades. Em relação as estações do ano, observou-se que no outono a espessura dos tecidos/estruturas são menores. Diante dos fatores genotóxicos e morfoanatômicos aqui avaliados, a microrregião da Bodoquena parece estar bem preservada em termos de qualidade do ar, apresentando baixa frequência de micronúcleos e redução limitada de tecidos e estruturas foliares, independentemente da estação do ano.

Abstract The objective of this study was to assess air quality in relation to vehicular traffic flow in cities located at different elevations in the Bodoquena microregion, state of Mato Grosso do Sul, Brazil. To do so, a micronucleus test was carried out using the TRAD-MCN bioassay on young Tradescantia buds collected from February to November 2018 in seven cities of the microregion with different traffic flow intensities. Meteorological parameters were evaluated, and vehicular traffic was counted to determine traffic flow in each city. With data from the Shuttle Radar Topography Mission (SRTM) and processing in Esri ArcGIS® software version 10.5.1, the regions was mapped based on an Elevation Model. Morphoanatomical analyses were performed according to standard methodology. Measurements were taken of thickness, length and width of tissues and structures, including the upper and lower cuticle, upper and lower epidermis, hypodermis and mesophyll. The greatest traffic flow was found in the cities of Bodoquena, Guia Lopes da Laguna, Jardim, and Porto Murtinho, with the period from 5:00 to 6:00 p.m. showing the highest traffic flow. The greatest frequency of mutagenic alterations was found in the city of Guia Lopes da Laguna, although the results did not differ significantly from Bonito, Caracol, and Jardim. Throughout the biomonitoring, the summer and autumn seasons showed the greatest micronuclei frequencies in all evaluated cities. Variations in the tissue/structure thickness was observed across cities and seasons, but with a decrease in thickness during autumn. In general, the tissues/structures were smaller for the cities of Nioaque and Porto Murtinho, while the anatomical and morphological characteristics of leaf length and thickness showed no differences among cities. We found limited correlation between micronuclei frequency and traffic flow, supporting the hypothesis that although mutagenic alterations are observed in T. pallida, in this microregion the changes are numerically lower when compared to other regions of the state. In light of the genotoxic and morphoanatomical factors assessed herein, the Bodoquena microregion appears to be well preserved in terms of air quality, presenting low micronuclei frequency and a limited reduction in tissues and leaf structures, regardless of the season.

ABSTRACT To evaluate the damage of oil exploration to ecosystems, ecotoxicological tests are a fundamental tool in environmental risk programs. The products used constitute xenobiotics that can be potentially toxic to soil biota and cause severe metabolic alterations in organisms, which affect structural and functional changes in terrestrial ecosystems. The objective of this research was to carry out a toxicological diagnosis of a water-based drilling fluid for use in oil exploration in Cuba. Toxicological procedures recommended by the Organization for Economic Cooperation and Development (OECD) were used, with two terrestrial contamination bioindicators, earthworms of the species Eisenia andrei and lettuce seeds (Lactuca sativa). The fluid was prepared by making a 1:9 dilutions (sludge: water) (v/v) and the dilutions to be tested were made from the fraction of suspended solids (FSS). As a toxicity criterion, the mean lethal concentration (LC50) was calculated from the mortalities of the bioindicators, with the Statgraphics Centurion XV statistical program, according to the Probit method. The values were compared with the current federal codes (CFR) that classify a toxic fluid when the LC50 is less than 3%. The LC50 for the bioassay with worms was 36.95% and 30.1% for lettuce seeds. According to established criteria, this fluid is non-toxic for the survival of terrestrial biota. These results support the use of this drilling fluid responsibly and with respect to the environment.

RESUMEN Para evaluar los daños de la exploración petrolera a los ecosistemas, los ensayos de ecotoxicológicos son una herramienta fundamental en los programas de riesgo ambiental. Los productos utilizados constituyen xenobióticos que pueden ser potencialmente tóxicos para la biota del suelo y ocasionar severas alteraciones metabólicas en los organismos, que repercuten en cambios estructurales y funcionales en los ecosistemas terrestres. El objetivo de esta investigación fue realizar un diagnóstico toxicológico a un fluido de perforación base agua para su uso en la exploración petrolera en Cuba. Se utilizaron procedimientos toxicológicos recomendadados por la Organización para la Cooperación Económica y el Desarrollo (OECD), con dos bioindicadores de contaminación terrestres, lombrices de tierra de la especie Eisenia andrei y semillas de lechuga (Lactuca sativa). El fluido se preparó realizando una dilución 1:9 (lodo: agua) (v/v) y a partir de la fracción de sólidos suspendidos (FSS) se realizaron las diluciones a ensayar. Como criterio de toxicidad se calculó la concentración letal media (CL50) a partir de las mortalidades de los bioindicadores, con el programa estadístico Statgraphics Centurion XV, según el método Probit. Los valores se compararon con los códigos federales (CFR) vigentes que cataloga a un fluido tóxico cuando la CL50 es menor que 3 %. La CL50 para el bioensayo con lombrices fue de 36.95 % y de 30.1 % para las semillas de lechuga; según los criterios establecidos este fluido es no tóxico para la supervivencia de la biota terrestre. Estos resultados apoyan el uso de este fluido de perforación con la responsabilidad y respecto al medio ambiente.

Abstract The methanolic, chloroformic and aqueous extract of Achillea millefolium and Chaerophyllum villosum were investigated for cytotoxicity, phytotoxic and insecticidal activities. Cytotoxicity was investigated by brine shrimp lethality assay indicating that the crude methanolic extract of A.millefolium and chloroformic extract of C.villosum revealed highest mortality of brine shrimps with (LD50 of 52.60 µg/ml) and (14.81 µg/ml). Phytotoxicity was evaluated using the Lemna minor bioassay which revealed that the crude methanolic extract of A.millefolium and C.villosum extract has maximum inhibition of Lemna minor with (Fl50 6.60 µg/ml) and (0.67 µg/ml).The insecticidal activity showed that among all the insects studied it was observed that methanolic extract of A. millefoliumand C. villosum was highly toxic to Sphenoptera dadkhani with (LD50=4.17 µg/ml) and (0.34 µg/ml). From the present study it can be concluded that different extracts from A. millefolium and C. villosum showed good cytotoxic, phytotoxic and insecticidal activity in a dose dependent manner. cytotoxicity activities Amillefolium A Cvillosum C LD50 LD (LD5 5260 52 60 52.6 µg/ml µgml µg ml 14.81 1481 14 81 (14.8 µg/ml. . Fl50 Fl (Fl5 660 6 6.6 0.67 067 0 67 (0.6 µg/ml.The µgmlThe .The LD50=4.17 LD50417 4 17 (LD50=4.1 0.34 034 34 (0.3 cytotoxic manner LD5 (LD 526 5 52. 14.8 148 1 8 (14. Fl5 (Fl 66 6. 0.6 06 (0. LD50=4.1 LD5041 (LD50=4. 0.3 03 3 14. (14 0. (0 LD50=4. LD504 (LD50=4 (1 ( LD50=4 (LD50= LD50=

Resumo Neste estudo, os extratos metanólico, clorofórmico e aquoso de Achillea millefolium e Chaerophyllum villosum foram analisados em relação à citotoxicidade, atividade fitotóxica e inseticida. A citotoxicidade foi analisada através do ensaio de letalidade de artémia, indicando que o extrato metanólico bruto de A. millefolium e o extrato clorofórmico de C. villosum revelaram maior mortalidade de artêmias com DL50 de 52,60 µg/ml e 14,81 µg/ml. A fitotoxicidade foi avaliada utilizando o bioensaio de Lemna minor que revelou que o extrato metanólico bruto de A. millefolium e extrato de C. villosum têm inibição máxima de Lemna minor com Fl50 6,60 µg/ml e 0,67 µg/ml. A atividade inseticida mostrou que dentre todos os insetos estudados, o extrato metanólico de A. millefolium e de C. villosum foi altamente tóxico para Sphenoptera dadkhanicom DL50 = 4,17 µg/ml e 0,34 µg/ml . Por outro lado, diferentes extratos, como A. millefolium e C. villosum apresentaram boa atividade citotóxica, fitotóxica e inseticida de forma dose-dependente. estudo artémia C DL DL5 5260 52 60 52,6 µgml µg ml 1481 14 81 14,8 Fl Fl5 660 6 6,6 067 0 67 0,6 estudados 417 4 17 4,1 034 34 0,3 lado citotóxica dosedependente. dosedependente dose dependente. dependente dose-dependente 526 5 52, 148 1 8 14, 66 6, 06 0, 41 4, 03 3

ABSTRACT The search for bioherbicides has been encouraged, and plants used in food or with bioactivity have been studied. Therefore, this article aimed to investigate the bioherbicidal potential of essential oils from Psidium cauliflorum and P. acidum through a plant toxicity bioassay using Lactuca sativa and Sorghum bicolor. The seeds were treated with essential oils of both species, along with control groups treated with distilled water, dichloromethane, and glyphosate. Germination percentage (GP), germination speed index (GSI), root growth (RG), shoot length (SL), mitotic index (MI), chromosomal alterations (CA), and nuclear alterations (NA) were evaluated. The major compound of the essential oil of P. cauliflorum was α-pinene, and of P. acidum were trans-caryophyllene, β-elemene, germacrene A, and α-copaene. The essential oils from both species exhibited phytotoxic effects. P. acidum oil inhibited sorghum RG and lettuce SL, while P. cauliflorum oil reduced GP, GSI, RG, and SL in both plants, indicating higher phytotoxicity than P. acidum and non-selective behavior. Cytotoxic investigations showed that both oils inhibited the MI. CA analysis revealed that P. cauliflorum oil exhibited aneugenic and clastogenic action mechanisms. The results demonstrate the bioherbicidal potential of P. cauliflorum essential oil, in addition to being non-selective and displaying a similar inhibition rate to glyphosate. encouraged studied Therefore P bicolor water dichloromethane glyphosate GP , (GP) GSI (GSI) (RG) (SL) MI, MI (MI) CA, (CA) NA (NA evaluated αpinene, Îpinene α pinene, Î pinene α-pinene transcaryophyllene, transcaryophyllene trans caryophyllene, caryophyllene trans-caryophyllene βelemene, βelemene Îelemene β elemene, elemene β-elemene A αcopaene. Îcopaene copaene. copaene α-copaene effects nonselective non selective behavior mechanisms (GP (GSI (RG (SL (MI (CA αpinene αcopaene

Resumo A mosca-branca é uma das pragas agrícolas mais difundidas no mundo. Os óleos essenciais podem ser usados para controlar esse inseto de forma ambientalmente responsável. A atividade fumigante, repelente e antioviposição de óleos essenciais extraídos com etanol de Trachyspermum ammi, Withania coagulans e Murraya koenigii contra Bemisia tabaci foi investigada neste estudo. No experimento, foram utilizadas três concentrações de óleo essencial (2,5 mg/mL, 5 mg/mL e 10 mg/mL). Trachyspermum ammi teve a maior porcentagem de morte de mosca-branca em experimentos de laboratório devido à sua toxicidade fumigante; a mesma tendência foi encontrada na toxicidade de contato e efeito repelente. A porcentagem de mortalidade aumenta à medida que a concentração de óleo essencial aumenta com o tempo do bioensaio. À medida que a concentração de óleo essencial cresce com o tempo de bioensaio, o mesmo acontece com a taxa de mortalidade. A maioria das moscas-brancas adultas escapou da estufa das plantas tratadas devido à maior concentração de óleo essencial. A maior repelência foi encontrada com óleo essencial de Trachyspermum ammi a 10 mg/mL. O óleo essencial apresentou a maior eficácia antioviposição contra moscas-brancas. Trachyspermum ammi teve o mais potente impedimento antioviposição, seguido por Withania coagulans em segundo lugar. Murraya koenigii terminou em terceiro com antioviposição moderada, o que afeta o número de ovos produzidos em relação ao controle. Como consequência, esses três óleos podem ser usados como um bioinseticida eficaz e ambientalmente aceitável para controlar B. tabaci.

Abstract Whitefly is one of the most widespread agricultural pests in the world. Essential oils might be used to control this insect in an environmentally responsible way. The fumigant, repellent, and anti-oviposition activity of ethanol-extracted essential oils of Trachyspermum ammi, Withania coagulans, and Murraya koenigii against Bemisia tabaci was investigated in this study. In the experiment, three essential oil concentrations (2.5 mg/mL, 5 mg/mL, and 10 mg/mL) were used. Trachyspermum ammi had the highest percentage of whitefly death in laboratory experiments due to its fumigant toxicity; the same tendency was found in contact toxicity and repellent effect. Mortality percent rises as the concentration of essential oil increases with bioassay time. As the concentration of essential oil grows with bioassay time, so does the mortality rate. The most adult whiteflies escaped from the treated plants' greenhouse due to the highest essential oil concentration. The greatest repellency was found with Trachyspermum ammi essential oil at 10 mg/mL. The essential oil had the greatest anti-oviposition efficacy against whiteflies. Trachyspermum ammi possessed the most potent anti-oviposition deterrent, followed by Withania coagulans in second place. Murraya koenigii finished third with moderate anti-oviposition, which affects the number of eggs produced in comparison to the control. As a consequence, these three oils might be used as an effective and environmentally acceptable bio-insecticide to control B. tabaci.

Abstract This study aimed to detect Toxoplasma gondii in artisanal salted meat products sold in street markets in the Ilhéus-Itabuna microregion and to assess the salt concentration used in their preparation and its influence on the parasite’s viability. A total of 125 samples of various artisanal meat products sold in street markets located in the Ilhéus-Itabuna microregion were collected during 2021. Serological analysis using indirect hemagglutination (HAI) and molecular analysis (PCR) were performed on these samples to detect the presence of the parasite. Möhr’s method was utilized to determine the sodium chloride concentration in the samples. Of all samples, 21 were subjected to a bioassay in albino mice to verify the viability of possible tissue cysts. Among the 125 meat products, 10 (8%) tested positive in the serological analysis including four cured pork sausages, five beef sun-dried meats, and one mixed fresh sausage (pork and chicken). None of 125 samples tested positive in the molecular analysis. On bioassay, all mice tested negative for the presence of the parasite. The NaCl concentration in the positive samples ranged from 2.9% to 8%. The results demonstrated that the salt concentration in the collected samples was sufficient to inactivate the parasite T. gondii. IlhéusItabuna Ilhéus Itabuna parasites s 12 2021 HAI (HAI PCR (PCR Möhrs Möhr 2 cysts 1 8% 8 (8% sausages sundried sun dried meats chicken. chicken . chicken) 29 9 2.9 T 202 (8 2. 20 (

Resumo Este estudo teve como objetivo detectar Toxoplasma gondii em produtos cárneos salgados artesanais, comercializados em feiras livres da microrregião Ilhéus-Itabuna, e avaliar a concentração do sal utilizado no preparo e sua influência na viabilidade do parasito. Foram coletadas 125 amostras de diversos produtos cárneos artesanais, vendidos em feiras livres localizadas na microrregião Ilhéus-Itabuna, durante o ano de 2021. Nessas amostras, a fim de detectar a presença do parasito, foram realizadas análises sorológicas de hemaglutinação indireta (HAI) e molecular (PCR). Para a determinação da concentração de cloreto de sódio presente nas amostras, foi realizado o Método de Möhr. Do total, 21 amostras foram destinadas ao bioensaio em camundongos albinos, na intenção de verificar a viabilidade de possíveis cistos teciduais presentes. Dos 125 produtos cárneos, 10 (8%) foram positivos na análise sorológica, sendo quatro linguiças curadas suína, cinco carnes de sol bovina e uma linguiça frescal mista (suíno e frango). Nenhuma amostra foi positiva na análise molecular. No bioensaio, todos os camundongos testaram negativo para a presença do parasito. Das amostras positivas na sorologia, as concentrações de NaCl variaram de 2,9% a 8%. Os resultados evidenciaram que a concentração de sal das amostras coletadas foi suficiente para inativar o parasito Toxoplasma gondii. artesanais IlhéusItabuna, IlhéusItabuna Ilhéus Itabuna, Itabuna Ilhéus-Itabuna 12 2021 HAI (HAI PCR. PCR . (PCR) Möhr total 2 albinos presentes 1 8% 8 (8% sorológica suína suíno frango. frango frango) sorologia 29 9 2,9 202 (PCR (8 2, 20 (

A series of biologically active N-acylamino amides were synthesized from readily available amino acids and common fatty acids from oil seeds, including the pharmacophoric group in the family of herbicides propanil, linuron, and diuron. The esterification followed by N-acylation with long-chain fatty acids was carried out with O-(benzotriazole-1-yl)-N,N,N’,N’-tetramethyluronium tetrafluoroborate (TBTU) as activating agent. The N-acylamino esters were synthesized and isolated in good yields (70-98%), which after the hydrolysis step, provided the N-acylamino acids. Finally, new N-acylamino amides were obtained from the reaction with 3,4-dichloroaniline in the presence of TBTU and N,N-diisopropylethylamine (DIPEA), in yields between 24-83%. Bioassays were conducted against arthropod strawberry pests Chaetosiphon fragaefolii, Duponchelia fovealis, and Drosophila suzukii, without any registered compound for use in strawberry crops in Brazil. The bioassay studies showed promising results, with mortality percentages ranging from 20-80%, and the best relative mortality by N-oleyl glycinamide, suggesting a possible fatty chain-amino acid synergistic bioactivity. Overall, this study provides an efficient and sustainable method for synthesizing biologically active N-acylamino amides from abundant natural sources, with potential applications in the development of ecofriendly agrochemicals. Nacylamino N acylamino seeds propanil linuron diuron Nacylation acylation longchain long chain Obenzotriazole1ylN,N,N,Ntetramethyluronium Obenzotriazole1ylNNNNtetramethyluronium ObenzotriazoleylNNNNtetramethyluronium O benzotriazole 1 yl N,N,N ,N tetramethyluronium (TBTU agent 7098%, 7098 70 98% , 98 (70-98%) step Finally 3,4dichloroaniline 34dichloroaniline dichloroaniline 3,4 3 4 N,Ndiisopropylethylamine NNdiisopropylethylamine N,N diisopropylethylamine DIPEA, DIPEA (DIPEA) 2483%. 2483 24 83%. 83 24-83% fragaefolii fovealis suzukii Brazil results 2080%, 2080 20 80%, 80 20-80% Noleyl oleyl glycinamide chainamino bioactivity Overall sources agrochemicals Obenzotriazole1ylN Ntetramethyluronium Obenzotriazole ylN ylNNNNtetramethyluronium NNN 7098% 709 7 9 (70-98% 4dichloroaniline 34 3, Ndiisopropylethylamine NN (DIPEA 2483% 248 2 83% 8 24-83 2080% 208 80% 20-80 ObenzotriazoleylN (70-98 24-8 20-8 (70-9 24- 20- (70- (70 (7 (

ABSTRACT Synthetic chemical insecticides are widely used in population control of pests. Aedes aegypti is the primary vector of dengue, Zika, chikungunya and yellow fever to humans, and has proven resistance to chemical insecticides. As an alternative vector control method, the ethanolic extract of the leaves of Piper peltatum L. (Piperaceae) showed larvicidal activity against Ae. aegypti. Despite the wide medicinal use of this plant, the biological activity of its isolated constituents remains unexplored. In this sense, we isolated, identified and evaluated the larvicidal activity of 4-nerolidylcatechol (4-NC) from P. peltatum against Ae. aegypti, Culex quinquefasciatus and Anopheles darlingi, focusing on the larvicidal, adulticidal and genotoxic potential of 4-NC on Ae. aegypti. Larvae were captured in the city of Manaus, Amazonas state, Brazil. 4-NC was isolated from the extract of the leaves of P. peltatum via silica gel chromatography. This was identified using nuclear magnetic resonance spectroscopy and tested in Artemia franciscana (6.25, 12.5, 25, 50, 100 and 200 µg/mL). In the toxicity bioassay, Ae. aegypti larvae were exposed to 30, 50, 70, 90, and 110 µg/mL of 4-NC, while Cx. quinquefasciatus and An. darlingi were exposed to 6.25, 12.5, 25, 50 and 100 µg/mL. Ae. aegypti larvae were also subjected to 40 and 60 µg/mL of 4-NC (genotoxicity bioassay), and adult females to 62.5 to 1,000 µg/mL (adulticidal bioassay). The results of the 4-NC toxicity assays showed that there was 100% mortality in larvae of Ar. franciscana at the concentration of 200 µg/mL, with an LC50 of 8.0 μg/mL. In the larvae of Ae. aegypti, mortality was 100%, with an LC50 of 62 μg/mL. In larvae of Cx. quinquefasciatus, 97% mortality occurred, with an LC50 of 52.3 μg/mL, and in An. darlingi larvae there was an 83% mortality rate, with an LC50 of 55.8 μg/mL. In adults of Ae. aegypti, however, there was no adulticidal activity. In the larvae of Ae. aegypti, the genotoxic effect of 4-NC (40 and 60 µg/mL) showed significant frequency (p < 0.05) of cellular abnormalities (micronuclei, budding and nuclear bridges) of interphasic nuclei of neuroblasts and oocytes in relation to the negative control. This result may be associated with a decrease in oviposition of females, which was observed in two generations. We can confirm that 4-NC has larvicidal activity against Ae. aegypti, Cx. quinquefasciatus and An. darlingi. Although it does not present adulticidal activity in Ae. aegypti, it reduced the oviposition of females. Therefore, 4-NC seems to be a strong candidate for the development of an alternative method for the control of these mosquitoes in the immature phase. pests dengue Zika humans L Piperaceae (Piperaceae Ae plant unexplored sense 4nerolidylcatechol nerolidylcatechol 4 4NC NC (4-NC P Manaus state Brazil chromatography 625 6 25 (6.25 125 12 5 12.5 10 20 µgmL . µg mL bioassay 30 70 90 11 4NC, NC, Cx An 6.25 genotoxicity , bioassay) 62. 1000 1 000 1,00 bioassay. Ar LC LC5 80 8 0 8. μgmL μg μg/mL 97 occurred 523 52 3 52. 83 rate 558 55 55. however (4 p 0.05 005 05 micronuclei, micronuclei (micronuclei bridges generations Therefore phase 2 (6.2 12. 7 9 6.2 00 1,0 ( 0.0 (6. 6. 1, 0. (6

Abstract Ctenocephalides felis felis is a relevant and widely distributed ectoparasite that acts as a vector of disease-causing pathogens. Moreover, it is responsible for economic losses due the use of harmful chemicals to the environment and that favor the emergence of insecticide-resistant populations. Research on entomopathogenic nematodes may open up an alternative route to the insect chemical control. The present study aimed to evaluate the killing efficacy of Heterorhabditis bacteriophora (HP88) against C. felis larvae in 400 μL, 600 μL and 1000 μL of suspension containing 120, 160 and 200 infective juveniles/larva and 600 μL of suspension containing the same concentrations of Heterorhabditis indica (LPP30), divided into two groups (absence and presence of diet) and a control group with three replications containing only distilled water. In the bioassay with H. bacteriophora, the groups in 600 μL of suspension showed higher mortality rates than those in the other tested volumes, which were above 80% at all concentrations. On the other hand, H. indica achieved mortality rates above 70% at all concentrations used. Results indicate that flea larvae are susceptible to in vitro infection by H. bacteriophora and H. indica. diseasecausing disease causing pathogens Moreover insecticideresistant insecticide resistant populations HP88 HP (HP88 C 40 60 100 120 16 20 juvenileslarva juveniles larva LPP30, LPP30 LPP , (LPP30) absence diet water H volumes 80 hand 70 used HP8 (HP8 4 6 10 12 1 2 LPP3 (LPP30 8 7 (HP (LPP3 (LPP

Resumo Ctenocephalides felis felis é um ectoparasito de relevância, ampla distribuição geográfica, vetor de patógenos causadores de doenças e responsável por perdas econômicas relacionadas ao uso de produtos químicos que causam impacto ambiental e aparecimento de populações resistentes. Pesquisas com nematoides entomopatogênicos podem ser uma alternativa ao controle químico desse inseto. O presente estudo objetivou avaliar a eficácia de Heterorhabiditis bacteriophora (HP88) em larvas de C. felis em 400 μL, 600 μL e 1000 μL de suspensão contendo 120, 160 e 200 juvenis infectantes/larva e 600 μL de suspensão contendo as mesmas concentrações de Heterorhabditis indica (LPP30), divididos em dois grupos (ausência e presença de dieta) e grupo controle com três repetições contendo apenas com água destilada. No bioensaio com H. bacteriophora observou-se que os grupos com 600 μL de suspensão apresentaram melhores percentuais de mortalidade em relação aos outros volumes testados, sendo estas acima de 80% para as três concentrações. Já H. indica apresentou percentuais de mortalidades acima de 70% para as três concentrações utilizadas. Os resultados indicam que as larvas de pulga são suscetíveis a infecção in vitro por H. bacteriophora e H. indica. relevância geográfica resistentes inseto HP88 HP (HP88 C 40 60 100 120 16 20 infectanteslarva infectantes larva LPP30, LPP30 LPP , (LPP30) ausência dieta destilada H observouse observou se testados 80 70 utilizadas HP8 (HP8 4 6 10 12 1 2 LPP3 (LPP30 8 7 (HP (LPP3 (LPP

Abstract Natural products, specifically plant extracts with biological activity and the ability to act as botanical biopesticides are often mistakenly considered nontoxic. Scientific evidence indicates the contrary, and for this reason, the objective of this work was to evaluate the toxicity of extracts obtained from Petiveria alliacea L. (Caryophyllales, Phytolaccaceae) using Daphnia magna Straus (Cladocera, Daphniidae) as a bioindicator to identify the plant extracts and the respective concentrations that present the highest toxicity. Leaves of P. alliacea were collected in the Peruvian amazone. From this material, three types of extract (hexane, ethanolic and aqueous) were prepared, which were used in the bioassays with D. magna to find the least toxic extract. Acute toxicity bioassays with D. magna during 48 h of exposure to hexane, ethanolic, and aqueous extracts yielded median lethal concentration (LC50) values of 26.9, 230.6, and 657.9 mg L-1, respectively. The aqueous extract presented the lowest toxicity, causing minimal D. magna mortality in the range of 6.67 to 13.33% at concentrations of 10 and 100 mg L-1. This result enables the efficient use of this plant species in a sustainable manner with a minimal environmental impact for the future development of natural products for pest control. nontoxic contrary reason L Caryophyllales, Caryophyllales (Caryophyllales Phytolaccaceae Cladocera, Cladocera (Cladocera Daphniidae P amazone material hexane (hexane prepared D 4 LC50 LC (LC50 269 26 9 26.9 2306 230 6 230.6 6579 657 657. L1, L1 1, 1 L-1 respectively 667 67 6.6 1333 13 33 13.33 L1. 1. control LC5 (LC5 2 26. 23 230. 65 L- 66 6. 133 3 13.3 (LC 13.

Resumo A utilização de produtos naturais, especificamente extratos vegetais com atividade biológica e capacidade de atuar como biopraguicidas botânicos, é muitas vezes erroneamente considerada não tóxica. Evidências científicas demonstram o contrário e por isso o objetivo deste trabalho consistiu na avaliação da toxicidade de extratos obtidos de Petiveria alliacea L. (Caryophyllales, Phytolaccaceae) com a utilização de Daphnia magna Straus (Cladocera, Daphniidae) como bioindicador, que permitiu descartar os extratos vegetais e as respectivas concentrações que apresentaram maior toxicidade. As folhas de P. alliacea foram coletadas na Amazônia peruana. A partir deste material, foram preparados três tipos de extrato − hexânico, etanólico e aquoso −, que foram utilizados nos bioensaios com D. magna a fim de encontrar o extrato menos tóxico. Os bioensaios de toxicidade aguda com D. magna durante 48 h de exposição aos extratos hexânico, etanólico e aquoso permitiram estabelecer os valores da Concentração Letal Média (CL50) de 26,9; 230,6 e 657,9 mg L-1, respectivamente. O extrato aquoso apresentou a menor toxicidade, causando percentuais mínimos de mortalidade de D. magna na faixa de 6,7 a 13,3% nas concentrações de 10 e 100 mg L-1. Estes resultados possibilitam o uso eficiente das espécies vegetais de forma sustentável e com mínimo impacto ambiental para o futuro desenvolvimento de produtos naturais para controle de pragas. botânicos tóxica L Caryophyllales, Caryophyllales (Caryophyllales Phytolaccaceae Cladocera, Cladocera (Cladocera Daphniidae bioindicador P peruana material hexânico D tóxico 4 CL50 CL (CL50 26,9 269 26 9 2306 230 6 230, 6579 657 657, L1, L1 1, 1 L-1 respectivamente 67 7 6, 133 13 3 13,3 L1. 1. pragas CL5 (CL5 26, 2 23 65 L- 13, (CL

ABSTRACT The horizontal transmission of the entomopathogenic fungus Beauveria bassiana Unioeste 76 strain among adults of Euschistus heros was evaluated in the laboratory and greenhouse using sporulated cadavers and live insects contaminated as a source of inoculate. Adult insects in the laboratory bioassay either received treatments with dry conidia or were sprayed with an oily dispersion formulation, and the treatments were comprised of 10, 30, or 50% of contaminated insects. For the greenhouse bioassay, nylon cages were prepared, with soybean plants grown in pots. A total of 20 healthy brown stink bugs were released along with one or three sporulated cadavers of brown stink bugs, attached to the plant stem. In the bioassay with live insects, 20 stink bugs were released, and the treatments were comprised of 10, 30, or 50% contaminated insects (2:18, 6:14, and 10:10, fungus-treated insects : healthy insects). In all experiments, the control group was comprised only of healthy insects. Dead stink bugs were daily disinfested and transferred to a wet chamber for confirmation of death by fungus. Confirmed mortality was observed, which shows the occurrence of horizontal transmission. Mortality was higher when more contaminated insects were used in the population, in most treatments. The occurrence of transmission in the laboratory and the greenhouse emphasizes the potential of the Unioeste 76 strain as mortality agent and as an agent for maintaining the inoculum potential for the brown stink bug. 7 inoculate formulation 10 30 50 prepared pots 2 stem 218, 218 18, 18 (2:18 614, 614 6 14, 14 6:14 1010, 1010 10:10 fungustreated treated . insects) experiments observed population bug 1 3 5 21 (2:1 61 6:1 101 10:1 (2: 6: 10: (2 (

Abstract Golden-Dewdrop (Duranta erecta L.), known for its landscaping applications, undergoes successive pruning to maintain its ornamental structure, a practice known to induce numerous physiological and chemical responses in the plant. Therefore, this study aimed to evaluate the influence of pruning and different extraction methods on the allelopathic potential of Golden-Dewdrop on the germination and initial growth of lettuce. Leaf extracts were obtained by decoction (hot) and maceration (cold) at concentrations of 20%, 40%, 60%, 80%, and 100%, with distilled water as a negative control. Phytochemical screening of the extracts was performed for phenolic compounds, flavonoids, anthocyanidins, tannins, cardioactive heterosides, saponins, alkaloids, and terpenes. For the bioassay, germinative and morphological parameters were measured and characterized. The data were subjected to analysis of variance (ANOVA), and for quantitative data, regression models were adjusted, while for qualitative data, means were compared using the Scott-Knott test with 5% significance. Significant differences were observed between extraction methods and management practices, with the hot aqueous extract from pruned plants significantly reducing germination at higher concentrations. The initial growth of lettuce was severely affected, presenting numerous morphological abnormalities, with severe damage to the root system at all concentrations. This study suggests that pruning practices have a modulating effect on the allelopathic activity of Golden-Dewdrop, providing valuable information for landscaping projects and botanical studies considering its evaluation and production of phytotoxic phytochemicals. GoldenDewdrop Golden Dewdrop Duranta L., L L. , L.) applications structure plant Therefore (hot cold (cold 20 20% 40 40% 60 60% 80 80% 100 100% control compounds flavonoids anthocyanidins tannins heterosides saponins alkaloids terpenes bioassay characterized ANOVA, ANOVA (ANOVA) adjusted ScottKnott Scott Knott 5 significance affected abnormalities GoldenDewdrop, Dewdrop, phytochemicals 2 4 6 8 10 (ANOVA 1

Resumo Pingo-de-ouro (Duranta erecta L.), conhecida por suas aplicações paisagísticas, é frequentemente submetida a sucessivas podas para manter sua estrutura ornamental, sabendo-se que essa prática gera inúmeras respostas fisiológicas e químicas na planta. Portanto, este estudo teve como objetivo avaliar a influência da poda e de diferentes métodos de extração sobre o potencial alelopático da Pingo-de-ouro na germinação e crescimento inicial da alface. Os extratos das folhas foram obtidos por decocção (quente) e maceração (frio) em concentrações de 20%, 40%, 60%, 80% e 100%, tendo água destilada como controle negativo. Foi realizada a triagem fitoquímica dos extratos para identificar compostos fenólicos, flavonoides, antocianidinas, taninos, heterosídeos cardiotônicos, saponinas, alcaloides e terpenos. Para o bioensaio, foram mensurados e caracterizados parâmetros germinativos e morfológicos. Os dados foram submetidos à análise de variância (ANOVA) e, para os dados quantitativos, foram ajustados modelos de regressão, enquanto para os dados qualitativos, as médias foram comparadas pelo teste de Scott-Knott com 5% de significância. Foram observadas diferenças significativas entre os métodos de extração e as práticas de manejo, sendo que o extrato aquoso quente das plantas podadas reduziu significativamente a germinação em concentrações mais elevadas. O crescimento inicial da alface foi gravemente prejudicado, apresentando inúmeras anormalidades morfológicas, com danos severos ao sistema radicular em todas as concentrações. Este estudo sugere que as práticas de poda têm um efeito modulador na atividade alelopática da Pingo-de-ouro, fornecendo informações valiosas para projetos paisagísticos e estudos botânicos, considerando sua avaliação e produção de fitoquímicos fitotóxicos. Pingodeouro Pingo ouro Duranta L., L L. , L.) paisagísticas ornamental sabendose sabendo se planta Portanto (quente frio (frio 20 20% 40 40% 60 60% 80 100 100% negativo fenólicos flavonoides antocianidinas taninos cardiotônicos saponinas terpenos bioensaio morfológicos ANOVA (ANOVA quantitativos regressão qualitativos ScottKnott Scott Knott 5 significância manejo elevadas prejudicado morfológicas Pingodeouro, ouro, botânicos fitotóxicos 2 4 6 8 10 1