RESUMEN
Rachiplusia nu Guenée is a polyphagous species able to develop on several cultivated and non-cultivated host plants. However, basic life history information about this pest on hosts is scarce. In this study, R. nu larvae did not survive on leaves of non-Bt corn, wheat, Bt cotton that expresses proteins Cry1Ac+Cry2Ab2 or on Intacta2 Xtend soybean that expresses the Cry1A.105/Cry2Ab2/Cry1Ac proteins. Rachiplusia nu showed a viable egg-to-adult biological cycle (54%-66.3%) on non-Bt soybean, sunflower, canola, vetch, Persian clover, alfalfa, bean, and forage turnip hosts, similar to larvae raised on the artificial diet. In addition, R. nu was unable to complete larval development on non-Bt cotton, and only 45.2% of R. nu larvae reached the pupal stage when fed leaves of intacta RR2 PRO soybean that expresses the Cry1Ac protein. Larval and pupal mass of surviving insects on Cry1Ac soybean leaves were also lower (larval: 0.104 g; pupal: 0.099 g) in relation to other food sources (larval: 0.165-0.189 g; pupal: 0.173-0.192 g). The total fecundity of R. nu on Cry1Ac soybean leaves was ≈65% lower in relation to other food sources. This fact caused ≈60% the net reproductive rate (Ro) and intrinsic rate of increase (rm) when compared to other food sources. Our findings indicate that the Cry1Ac soybean negatively affects the biological parameters of R. nu. Non-Bt soybean, sunflower, canola, vetch, Persian clover, alfalfa, bean, and forage turnip are viable food sources for the survival and development of R. nu.
Asunto(s)
Toxinas de Bacillus thuringiensis , Endotoxinas , Proteínas Hemolisinas , Larva , Mariposas Nocturnas , Animales , Mariposas Nocturnas/crecimiento & desarrollo , Larva/crecimiento & desarrollo , Larva/fisiología , Dieta , Femenino , Pupa/crecimiento & desarrollo , Plantas Modificadas Genéticamente , Proteínas Bacterianas , Masculino , Tablas de Vida , Rasgos de la Historia de VidaRESUMEN
INTRODUCTION: Caffeine is a widely consumed substance with several effects on bone metabolism. This study aimed to investigate the effect of caffeine on the bone tissue of rats submitted to orthodontic movement. METHODS: Twenty-five male Wistar rats underwent orthodontic movement (21 days) of the first permanent maxillary molars on the left side. The experimental group (caffeine; n = 13) and control group (n = 12) received caffeine and water, respectively, by gavage. Microcomputed tomography was performed to analyze orthodontic movement. Histologic analysis of the inflammatory infiltrate and osteoclast count by tartrate-resistant acid phosphatase were conducted. Maxilla tissue was evaluated for receptor activator of nuclear factor Ò¡B (RANK), RANK ligand (RANKL), and osteoprotegerin by immunohistochemistry. RESULTS: Caffeine exhibited a lower bone volume/tissue volume ratio (78.09% ± 5.83%) than the control (86.84% ± 4.89%; P <0.05). Inflammatory infiltrate was increased in the caffeine group compared with the control group (P <0.05). A higher number of tartrate-resistant acid phosphatase-positive cells was observed in the caffeine (9.67 ± 1.73) than in the control group (2.66 ± 0.76; P <0.01). Immunoexpression of RANK and RANKL in the caffeine group was greater than the control (P <0.05). CONCLUSIONS: The use of caffeine thermogenic induces alveolar bone loss in rats submitted to orthodontic movement via activation of RANK, RANKL, and osteoprotegerin signaling pathways.
Asunto(s)
Pérdida de Hueso Alveolar , Cafeína , Osteoprotegerina , Ligando RANK , Receptor Activador del Factor Nuclear kappa-B , Técnicas de Movimiento Dental , Animales , Masculino , Ratas , Pérdida de Hueso Alveolar/metabolismo , Pérdida de Hueso Alveolar/patología , Cafeína/farmacología , Maxilar/efectos de los fármacos , Osteoclastos/efectos de los fármacos , Osteoprotegerina/metabolismo , Ligando RANK/metabolismo , Ratas Wistar , Receptor Activador del Factor Nuclear kappa-B/metabolismo , Transducción de Señal/efectos de los fármacos , Técnicas de Movimiento Dental/efectos adversos , Microtomografía por Rayos XRESUMEN
The exogenous application of bioregulators, such as salicylic acid (SA), has exhibited promising outcomes in alleviating drought stress. Nevertheless, its impact on culantro (Eryngium foetidum L.) remains unexplored. Thus, the aim of this study was to assess how SA impacts the growth, morphophysiology, and essential oil composition of culantro when subjected to drought. To achieve this, culantro plants were grown under three different watering regimes: well-watered, drought-stressed, and re-watered. Additionally, they were either treated with SA (100 µM) or left untreated, with water serving as the control. SA application did not mitigate the effects of drought in biomass production but increased biomass, leaf number, leaf area, and photosynthetic pigments under well-irrigated and re-watered conditions. After a drought period followed by re-watering, plants recovered membrane integrity independently of SA application. Water stress and the exogenous application of SA also modulated the profile of essential oils. This is the first report about SA and drought affecting growth and essential oil composition in culantro.
RESUMEN
As doenças crônicas não transmissíveis mais prevalentes no mundo são as doenças cardiovasculares, diabetes mellitus, alguns tipos de câncer e doenças respiratórias crônicas. Juntas essas condições foram responsáveis por mais da metade das mortes ocorridas no mundo e no Brasil e constituem o principal problema de saúde. As práticas de autocuidado com a saúde têm sido apontadas como a parte que cabe ao paciente nas estratégias de promoção da saúde. Alguns estudos vêm abordando a resiliência como um importante indicador para a realização de práticas saudáveis de autocuidado, estando associada à autonomia e às habilidades para a autogestão da saúde. O objetivo deste estudo é analisar a associação entre a resiliência psicológica e o autocuidado com a saúde em mulheres adultas. O estudo foi realizado com 159 pacientes dos ambulatórios de ginecologia do Instituto Nacional de Saúde da Mulher, da Criança e do Adolescente Fernandes Figueira, da Fundação Oswaldo Cruz. Foi utilizada a versão brasileira da Escala de Resiliência de Wagnild & Young e um questionário adaptado do Inquérito Vigilância de Fatores de Risco e Proteção para Doenças Crônicas por Inquérito Telefônico. O principal resultado foi importância do contexto familiar para os níveis de resiliência, principalmente para mulheres que têm filhos. Não foram encontradas associações significativas entre resiliência e as variáveis de autocuidado. Novas pesquisas deverão ser desenvolvidas para esclarecer as possíveis relação entre resiliência e autocuidado e a importância da informação em saúde para promover mudanças de estilo de vida.
The most prevalent chronic non-communicable diseases in the world are cardiovascular diseases, diabetes mellitus, some types of cancer and chronic respiratory diseases. Together, these conditions were responsible for more than half of the deaths in the world and in Brazil, they constitute the main health problem. Health self-care practices have been identified as the patient's part in health promotion strategies. Some studies have approached resilience as an important indicator for healthy self-care practices, being associated with autonomy and skills for self-management of health. The aim of this study is to analyze the association between psychological resilience and self-care with health in adult women. The study was carried out with 159 patients from the gynecology outpatient clinics of the Fernandes Figueira National Institute of Women's, Children and Adolescents' Health of the Oswaldo Cruz Foundation. The Brazilian version of the Wagnild & Young Resilience Scale and a questionnaire adapted from the Surveillance of Risk and Protection Factors for Chronic Diseases by Telephone Survey were used. The main result was the importance of the family context for resilience levels, especially for women who have children. No significant associations were found between resilience and self-care variables. New research should be developed to clarify the possible relationship between resilience and self-care and the importance of health information to promote lifestyle changes.
Asunto(s)
Humanos , Femenino , Autocuidado , Enfermedad Crónica/prevención & control , Salud de la Mujer , Resiliencia Psicológica , Atención Ambulatoria , Enfermedades no Transmisibles/prevención & control , Promoción de la Salud , BrasilRESUMEN
Climacteric is the period in a woman's life cycle that involves perimenopause, menopause and postmenopause. It represents the end of the reproductive period and the beginning of female aging in the biological sense. Over the past 10 years, an emphasis has been placed on the study of mental health in the female climacteric. Studies about the prevalence of depression, anxiety, suicidal ideation and association with different factors have shown important results on the relevance of these disorders during female climacteric. Considering the importance of these findings in the field of public health and epidemiology, we conducted a narrative review aiming to discuss the current knowledge about the prevalence of depression, anxiety and suicidal ideation in the female climacteric, analyzing methods and results in different studies, selected from a literature review between 2009 and 2019, in the PubMed, Lilacs, Embase, Web of Science and Google Scholar databases. Our results showed that the methodology, cultural and sociodemographic differences, as well as a variety of biopsychosocial factors studied, generate some uncertainties about the exact relationship between depression, anxiety, suicidal ideation and female climacteric. We conclude that more research, greater methodological rigor, more accurate results and a biopsychosocial view are needed and urgent for effective interventions, cost reduction in mental health care, as well as prevention and control of depression, anxiety and suicide in climacteric.
Asunto(s)
Ansiedad , Suicidio , Climaterio , Depresión , MujeresRESUMEN
The objectives were (a) to evaluate whether grasses native to the Pampa biome, Axonopus affinis Chase, Paspalum notatum Flüggé and Paspalum plicatulum Michx, and the invasive grass Cynodon dactylon (L.). Pers have the potential to phytoremediate soil contaminated with Cu (0, 35 and 70 mg Cu kg-1); (b) assess whether the growth of these species is compromised by the excess of Cu available in the soil; and (c) determine the impact of excess Cu on the physiological responses of the studied species. C. dactylon presented the best performance in soil contaminated with 35 mg of Cu kg-1. In C. dactylon, the concentrations of chlorophyll b and carotenoids increased, as did the photosynthetic rate and plant growth. Phytotoxic effects of Cu in soil contaminated with 70 mg of Cu kg-1 were more severe on A. affinis and led to plant death. The other species presented reduced photosynthetic and growth rates, as well as increased activity of antioxidant enzymes such as superoxide dismutase and guaiacol peroxidase. This very same Cu level has decreased photosynthetic pigment concentrations in P. notatum and P. plicatulum. On the other hand, it did not change chlorophyll a and b concentrations in C. dactylon and increased carotenoid concentrations in it. High values recorded for Cu bioaccumulation-in-grass-root factor, mainly in P. plicatulum, have indicated that the investigated plants are potential phytostabilizers. High C. dactylon biomass production-in comparison to other species-compensates for the relatively low metal concentration in its tissues by increasing metal extraction from the soil. This makes C. dactylon more efficient in the phytoremediation process than other species.
Asunto(s)
Paspalum , Contaminantes del Suelo , Biodegradación Ambiental , Granjas , Contaminantes del Suelo/análisis , Clorofila A , Especies Introducidas , Brasil , Cobre/análisis , Suelo , Ecosistema , AntioxidantesRESUMEN
Water deficit is the most critical factor limiting plant growth and production and salicylic acid (SA) has potential for stress mitigation in plants; therefore, we evaluated the effect of SA on radish (Raphanus sativus L.) growth and ecophysiology under water deficit. Plants were sprayed with SA (100µM) or water (control), and irrigated at 80% (W80), 60% (W60), 40% (W40), and 20% (W20) of field capacity. The SA treatments and drought stress started 7days after sowing and lasted until the end of the cycle (30days after sowing). The morphophysiological analyses showed that radish plants had impaired growth at the lower water supply levels, but the treatment with SA reversed these growth restraints under moderate stress, leading to increases in shoot mass at W40 and storage root mass at W60 and W40. SA treatment also reversed the reduction of storage root volume at W60. The tendency of water deficit to increase F O and reduce F V /F M suggests possible damage to the photosystem II of drought-stressed plants. The parameters of gas exchange and photosynthetic pigments showed maintained photosynthetic efficiency, but total photosynthesis decreased due the lower shoot dry mass. Overall, exogenously applied SA reversed the growth restraints at W60 and W40, which revealed that SA was effective in mitigating the effects of moderate water deficit on biomass accumulation and partitioning in radish plants.
Asunto(s)
Raphanus , Sequías , Fotosíntesis , Ácido Salicílico/farmacología , Agua/farmacologíaRESUMEN
ABSTRACT: The traffic in international animal products can become a public health hazard when legal import sanitary procedures are not followed. In Brazil, due to its extensive border area, the importation of animal products is a common practice in many areas, especially in Rio Grande do Sul, a state that borders Argentina and Uruguay. The objective of this study was to evaluate the presence of veterinary drug residues (antibiotics and antiparasitics) in animal products consumed in Rio Grande do Sul. The presence of residues of veterinary antibiotics and antiparasitics was assessed in 189 meat (beef, pork, and chicken), processed dairy, and meat product samples bought in Argentina (n = 90) and Uruguay (n = 99). Residues of these veterinary drugs were detected in 50 (26.45%) of the samples; 28 samples (14.81%) had antibiotic residues, and 22 samples (11.64%) had antiparasitic residues. Of the 50 positive samples, 40% (15 from Argentina and 5 from Uruguay) had residues above the maximum residue limits (MRLs). Of these 20 samples, 12 had antiparasitic residues above the MRLs (11 beef samples had ivermectin and 1 pork sample had ivermectin and doramectin) and 8 had antibiotic residues above the MRLs (2 pork and 2 sausage samples had doxycycline, 2 cheese samples had doxycycline and chlortetracycline, 1 poultry meat sample had chloramphenicol, and 1 cheese sample had monensin). Because of the potential toxic effects on humans and the potential for pathogens to develop antibiotic resistance, the presence of these residues above the MRLs is a potential risk to public health. The negative impact of consumption of imported animal products can be reduced by implementation of an effective surveillance system and educational campaigns for the general population.
Asunto(s)
Antiinfecciosos , Residuos de Medicamentos , Drogas Veterinarias , Animales , Antibacterianos/análisis , Antiparasitarios , Argentina , Brasil , Bovinos , Doxiciclina , Residuos de Medicamentos/análisis , Contaminación de Alimentos , Humanos , Ivermectina , UruguayRESUMEN
The aim of this study was to investigate the presence of adenovirus (AdV), rotavirus (RV), and hepatitis E virus (HEV) in beef, pork, and chicken meat cuts in retail trade in the city of Uruguaiana, RS, Brazil. A total of 131 meat products were collected (beef, n = 55; chicken, n = 47; pork, n = 29) from 18 commercial establishments (supermarkets, n = 7; butchers, n = 7; markets/grocery stores, n = 4). All samples were evaluated for AdV, RV, and HEV. The genomes of RV and AdV were identified in 29% (n = 38) and 5.34% (n = 7) of the samples, respectively. HEV was not identified in any of the samples. Chicken cuts had a higher frequency of AdV and RV isolates compared to beef and pork (P < 0.05). Among the categories of commercial establishments evaluated, all revealed at least one positive sample for AdV and RV; however, supermarkets showed a higher frequency of RV than others (P < 0.05). The genetic material of AdV and RV was identified simultaneously in 2.29% (n = 3) of samples from supermarkets (n = 2) and grocery stores (n = 1). This is the first report on detection of enteric viruses in meat cuts in the western region of the state of Rio Grande do Sul, Brazil, and the presence of AdV and RV in these products may indicate flaws during the process of handling these foods, especially in places where commercialization provides important public health issues.
RESUMEN
Using an online questionnaire, this study evaluated the profile of a Brazilian population's food handling practices in the home environment. The questionnaire, containing questions about domestic behavior in terms of hygiene and food handling, was built and available through social media sites. Information about the participants' profiles, their food pre-preparation, food preparation, and food post-preparation practices, and the occurrence of foodborne diseases (FBDs) was included in the questionnaire. A total of 701 responses were obtained. The interviewees included 78.31% female participants and 21.68% male participants, with an average age of 31.2 years. Nearly all (94.3%) had a complete or incomplete higher education. In the pre-preparation stage, the participants evaluated the shelf life (97.28%) and storage temperature (44.79%) of the products while purchasing them. Regarding food handling practices, only a few participants washed the food packages before storing them (31.95%) or removed hand jewelry or other adornments when washing food (61.48%). Most participants washed their hands (91.58%) and washed vegetables (99.28%). But a group of interviewees reported washing raw meat (27.81%) before preparing it. Cutting surfaces such as plastic (50.36%) and glass (49.36%) tops were the most prevalent in the study. Most respondents did not know how long they had been using their cutting boards (67.62%) and mentioned using the same surface to handle both raw and ready-to-eat products (84.17%). As for the preparation, most interviewees declared they did not check the food temperature during preparation (86.31%), ignoring the ideal cooking temperature (88.26%). Regarding the occurrence of FBDs, 79.17% of the interviewees reported having suspicious clinical signs associated with contaminated foods and 65.59% did not seek medical help. Thus, the participants demonstrated ignorance about adequate practices for food safety in the home environment, highlighting the need to conduct health education programs within the Brazilian population.(AU)
Este estudo teve como objetivo avaliar o perfil das práticas de manipulação de alimentos no ambiente domiciliar no Brasil utilizando um questionário online. Um questionário contendo perguntas sobre comportamento doméstico em nível de higiene e manipulação de alimentos foi construído e disponibilizado por redes sociais. O questionário continha informações sobre o perfil dos participantes, suas práticas de pré-preparo, preparo e pós-preparo de alimentos e a ocorrência de doenças transmitidas por alimentos (DTA). Obteve-se 701 respostas, os entrevistados foram 78,31% do sexo feminino e 21,68% do sexo masculino, com média de idade de 31,2 anos. A maioria (94,3%) possuia ensino superior completo ou incompleto. Na etapa de pré-preparo, os participantes avaliam o prazo de validade (97,28%) e a temperatura de armazenamento (44,79%) dos produtos no momento da compra. Em relação às práticas de manipulação dos alimentos, apenas alguns participantes lavavam as embalagens dos alimentos antes de armazená-los (31,95%) ou retiravam adornos ao lavar os alimentos (61,48%). A maioria dos participantes lavam as mãos (91,58%) e os vegetais (99,28%); entretanto, um grupo de entrevistados relatou lavar carne crua (27,81%) antes de prepará-la. Superfícies de corte como tábuas de plástico (50,36%) e de vidro (49,36%) foram os mais prevalentes no estudo. A maioria dos entrevistados não sabe há quanto tempo usa as tábuas de corte (67,62%) e utilizam a mesma superfície para manusear produtos crus e prontos para o consumo (84,17%). Quanto ao preparo, a maioria dos entrevistados declarou não verificar a temperatura dos alimentos durante o preparo (86,31%), ignorando a temperatura ideal de cozimento (88,26%). Em relação à ocorrência de DVA, 79,17% dos entrevistados relataram que já apresentaram sinais clínicos suspeitos associados a alimentos contaminados e 65,59% não procuraram atendimento médico. Nesse sentido, os participantes demonstraram desconhecimento sobre as práticas adequadas para a segurança dos alimentos no ambiente domiciliar, evidenciando a necessidade de realização de programas de educação em saúde com a população brasileira.(AU)
Asunto(s)
Animales , Masculino , Femenino , Encuestas de Atención de la Salud/instrumentación , Manipulación de Alimentos/estadística & datos numéricos , Abastecimiento de Alimentos/estadística & datos numéricos , Brasil , Higiene Alimentaria/estadística & datos numéricos , Buenas Prácticas de Manipulación , Enfermedades Transmitidas por los Alimentos/prevención & controlRESUMEN
Resumo As mudanças demográficas associadas à complexidade dos problemas ambientais contemporâneos, como as mudanças ambientais globais e os desastres tecnológicos, tornarão cada vez mais perene a (re)produção social dos riscos e desastres a eles associados. O artigo propõe reflexões que posicionem a demografia, particularmente no contexto brasileiro, de forma a incorporar esses desafios aos seus conceitos, teorias e metodologias de análise, consolidando o campo de estudos em demografia dos desastres. O percurso escolhido foi, inicialmente, o de revisitar conceitos presentes em estudos de população e ambiente, como riscos, danos, desastres, vulnerabilidade, adaptação e resiliência. Revisitamos a literatura produzida em demografia dos desastres, enfatizando a relação endógena entre desastres e a composição, distribuição e dinâmica demográfica. Em seguida, propusemos um marco teórico sobre demografia dos desastres, bem como sua operacionalização a partir de sete princípios. Por fim, discutimos como, tanto do ponto vista conceitual, teórico quanto metodológico, a demografia possui um papel fundamental para consolidar uma perspectiva científica que antagonize discursos de "naturalização" dos desastres e, consequentemente, contribua para criar ou aperfeiçoar políticas públicas e mecanismos de gestão e planejamento antes, durante e após os desastres.
Abstract Demographic changes, associated with the complexity of contemporary environmental problems such as global environmental changes and technological disasters, will make the social (re)production of risks and associated disasters increasingly permanent. The article proposes reflections that position Demography, particularly in the Brazilian context, in order to incorporate these challenges into its concepts, theories, analyses and methodologies, consolidating the field of knowledge in Demography of Disasters. The strategy initially chosen was to revisit concepts in population and environment studies, such as risks, damages, disasters, vulnerability, adaptation and resilience. We reviewed the literature produced on Demography of Disasters, emphasizing the endogenous relationship between disasters and demographic composition, distribution and dynamics. Then, we proposed a theoretical framework on Demography of Disasters, and its operationalization in seven principles. Finally, we discuss how, both from a conceptual and theoretical point of view, as well as from a methodological standpoint, demography plays a fundamental role in consolidating a scientific perspective that antagonizes discourses of "naturalization" of disasters and, consequently, contributes to creating or improving public policies and management and planning before, during and after disasters.
Resumen Los cambios demográficos, asociados a la complejidad de los problemas ambientales contemporáneos como los cambios ambientales globales y los desastres tecnológicos, harán que la (re)producción social de los riesgos y desastres asociados sea cada vez más permanente. El artículo propone reflexiones que posicionan a la Demografía, particularmente en el contexto brasileño, de forma que pueda incorporar estos desafíos en sus conceptos, teorías y metodologías, consolidando el campo de estudios en la Demografía de los desastres. El camino escogido inicialmente fue el el revisitar conceptos en estudios de población y medioambiente, como riesgos, daños, desastres, vulnerabilidad, adaptación y resiliencia. Se revisó la literatura producida sobre Demografía de los desastres, con énfasis en la relación endógena entre los desastres y la composición, distribución y dinámica demográfica. Luego se propuso un marco teórico sobre Demografía de desastres y su operacionalización a partir de siete principios. Finalmente, se discutió cómo, tanto desde el punto de vista conceptual y teórico como desde el de las metodologías de evaluación, la Demografía juega un papel fundamental en la consolidación de una perspectiva científica que antagoniza los discursos de naturalización de los desastres y, en consecuencia, contribuye a crear o mejorar las políticas públicas y los mecanismos de gestión y planificación antes, durante y después de los desastres.
Asunto(s)
Humanos , Brasil , Demografía , Desastres , InvestigaciónRESUMEN
Highbush (Vaccinium corymbosum L.) and rabbiteye (V. ashei R.) blueberry are the most important export small fruit crops in southern Brazil. Anthracnose has been considered one of the most destructive disease and exclusively associated with C. karstii in Brazil (Rios et al. 2014). In November 2019, severe anthracnose symptoms including leaf spots but particularly twig blights and fruit rots were observed on all blueberry plants (V. ashei) in one organic orchard in Santa Catarina state, Brazil (27º43'48.96"S, 49º0'57.79"W). Four isolates were obtained from necrotic lesions and monosporic cultures were grown on potato dextrose agar at 25°C and with a 12 h photoperiod under near ultra violet light. After 15 days, colonies showed upper surface color varying from grayish-white to pale-orange and the reverse side pale-orange. Conidia were hyaline, cylindrical with rounded ends, and their length and width ranged from 9.5 to 15.5 µm (x Ì =11.8) and 6.5 to 3.5 µm (x Ì =4.9), respectively. The isolates were identified by multilocus phylogenetic analyses using nucleotide sequences of actin (ACT), ß-tubulin (TUB2), calmodulin (CAL), glyceraldehyde-3-phosphate-dehydrogenase (GAPDH), glutamine synthetase (GS), internal transcribed spacer (ITS) and the intergenic spacer between DNA lyase and the mating-type locus MAT1-2-1 (ApMAT). Nucleotide sequences exhibited from 95 to 100% sequence identity to Colletotrichum chrysophilum ex-type (CMM4268) and were deposited in GenBank database (MW868219 to MW868222, MW868211 to MW868214, MW868215 to MW868218, MW868223 to MW868226, MW868202 to MW868205, MW793353 to MW793356, and MW868207 to MW868210). C. chrysophilum belongs to the C. gloeosporioides species complex and was previously described as C. ignotum in banana and other tropical fruits in Brazil (Vieira et al. 2017; Veloso et al. 2018). In addition, this species was recently reported on apple fruit in New York, USA (Khodadadi et al. 2020). To confirm pathogenicity, one-year-old blueberry plants were inoculated by spraying a suspension of 1×106 conidia/ml, incubated in a moist chamber in the dark for 48 h and then kept in the greenhouse. Plants sprayed with sterile distilled water served as control. Additionally, fruits were immersed for 2 min in a conidial suspension (1×106 conidia/ml) and incubated at 25°C and 12 h photoperiod for 20 days. Inoculated plants exhibited first symptoms in twigs at 10 days after inoculation (dai). Infected twigs showed initially dark brown spots that coalesced and became necrotic. On leaves, reddish-brown lesions with less than 2 mm appeared at low intensity at 15 dai. On fruits, sunken areas associated with an abundant orange mucilaginous mass of acervuli and conidia were seen at 7 dai. Symptoms on plants were identical to those observed under field conditions, and the pathogen was re-isolated from lesions fulfilling Koch's postulates. To the best knowledge, this is the first report of C. chrysophilum causing anthracnose on blueberries in Brazil. The identification of this species causing blueberry anthracnose is crucial to improve the disease control strategies and resistance breeding.
RESUMEN
Strawberry (Fragaria x ananassa Duch.) is one of the most important fruit crops worldwide. With increasing cultivated area in the last decades, Brazil has become the largest strawberry producer in South America. Anthracnose caused by Colletotrichum spp. has been considered one of the most destructive diseases in Brazil. In May 2019, irregular and circular dark brown leaf spots sometimes associated with chlorosis and petiole necrosis were observed on strawberry plants (cv. Pircinque) organically cultivated in Santa Catarina state, Brazil (27°45'40"S, 49°59'06''W). The Colletotrichum isolate was obtained from leaf, and monosporic culture was grown on potato dextrose agar at 25°C and 12-h photoperiod under near ultraviolet light. Colonies at the age of 15 days showed upper surface color varying from white to orange and the reverse side grayish to orange. Conidia were hyaline, cylindrical with rounded ends, 13.9 to 9.2 × 4.2 to 6.7 µm ((x ) Ì = 11.3 × 5.2, n = 100). Perithecia were produced in vitro and their diameter ranged from 265.2 to 142.5 µm ((x ) Ì = 198.4). Asci were 47.3 to 39.9 × 5.2 to 7.2 µm ((x ) Ì = 42.8 × 5.9, n = 50), and ascospores 12.6 to 8.1 × 4.3 to 2.1 ((x ) Ì = 10.3 × 2.9, n = 100). To confirm pathogenicity, 90-day-old plants of strawberry (cv. Pircinque) were inoculated by spraying a suspension of 1×106 conidia/ml, incubated in a moist chamber in the dark for 48 h and then kept in a greenhouse for further 30 days. Plants sprayed with sterile distilled water served as control. Additionally, detached leaves were inoculated with six drops of 10 µl (1×106 conidia/ml) onto abaxial surface and incubated in a moist chamber at 25°C and 12 h photoperiod for 15 days. Inoculated plants exhibited first symptoms in both leaves and petioles at 15 days after inoculation (dai). On leaf, irregular and circular dark brown spots evolved to necrotic lesions and were frequently surrounded by chlorotic halos. In petioles, lesions were reddish-brown, elongated, and depressed. Typical anthracnose symptoms on fruits at 6 dai showed as circular, slightly sunken lesions that enlarged over time and produced an abundant orange mucilaginous mass of acervuli and conidia, and after 20 days, fruits became mummified. In the detached-leaf-assay, symptoms appeared at 7 dai, with presence of circular dark brown lesions measuring 1 to 15 mm and then evolved to necrosis. The same pathogen was consistently re-isolated from the inoculated leaves, petioles, and fruits, and confirmed by morphological characterization and molecular assays as described in this note. A representative isolate (MANE189) was molecularly identified using genomic regions of actin (ACT), ß-tubulin (TUB2), calmodulin (CAL), glyceraldehyde-3-phosphate-dehydrogenase (GAPDH), glutamine synthetase (GS), and internal transcribed spacer (ITS). Nucleotide sequences exhibited 100% homology to the typical Colletotrichum karstii strains (CBS:127535, CBS:128500 and ML1792) and were deposited in GenBank database (MW396420, MW396430, MW396460, MW396440, MW396450, and MW331606). This species belongs to the C. boninense species complex (Damm et al. 2012) and was previoStrawberry (Fragaria x ananassa Duch.) is one of the most important fruit crops worldwide. With increasing cultivated area in the last decades, Brazil has become the largest strawberry producer in South America. Anthracnose caused by Colletotrichum spp. has been considered one of the most destructive diseases in Brazil. In May 2019, irregular and circular dark brown leaf spots sometimes associated with chlorosis and petiole necrosis were observed on strawberry plants (cv. Pircinque) organically cultivated in Santa Catarina state, Brazil (27°45'40"S, 49°59'06''W). The Colletotrichum isolate was obtained from leaf, and monosporic culture was grown on potato dextrose agar at 25°C and 12-h photoperiod under near ultraviolet light. Colonies at the age of 15 days showed upper surface color varying from white to orange and the reverse side grayish to orange. Conidia were hyaline, cylindrical with rounded ends, 13.9 to 9.2 × 4.2 to 6.7 µm ((x ) Ì = 11.3 × 5.2, n = 100). Perithecia were produced in vitro and their diameter ranged from 265.2 to 142.5 µm ((x ) Ì = 198.4). Asci were 47.3 to 39.9 × 5.2 to 7.2 µm ((x ) Ì = 42.8 × 5.9, n = 50), and ascospores 12.6 to 8.1 × 4.3 to 2.1 ((x ) Ì = 10.3 × 2.9, n = 100). To confirm pathogenicity, 90-day-old plants of strawberry (cv. Pircinque) were inoculated by spraying a suspension of 1×106 conidia/ml, incubated in a moist chamber in the dark for 48 h and then kept in a greenhouse for further 30 days. Plants sprayed with sterile distilled water served as control. Additionally, detached leaves were inoculated with six drops of 10 µl (1×106 conidia/ml) onto abaxial surface and incubated in a moist chamber at 25°C and 12 h photoperiod for 15 days. Inoculated plants exhibited first symptoms in both leaves and petioles at 15 days after inoculation (dai). On leaf, irregular and circular dark brown spots evolved to necrotic lesions and were frequently surrounded by chlorotic halos. In petioles, lesions were reddish-brown, elongated, and depressed. Typical anthracnose symptoms on fruits at 6 dai showed as circular, slightly sunken lesions that enlarged over time and produced an abundant orange mucilaginous mass of acervuli and conidia, and after 20 days, fruits became mummified. In the detached-leaf-assay, symptoms appeared at 7 dai, with presence of circular dark brown lesions measuring 1 to 15 mm and then evolved to necrosis. The same pathogen was consistently re-isolated from the inoculated leaves, petioles, and fruits, and confirmed by morphological characterization and molecular assays as described in this note. A representative isolate (MANE189) was molecularly identified using genomic regions of actin (ACT), ß-tubulin (TUB2), calmodulin (CAL), glyceraldehyde-3-phosphate-dehydrogenase (GAPDH), glutamine synthetase (GS), and internal transcribed spacer (ITS). Nucleotide sequences exhibited 100% homology to the typical Colletotrichum karstii strains (CBS:127535, CBS:128500 and ML1792) and were deposited in GenBank database (MW396420, MW396430, MW396460, MW396440, MW396450, and MW331606). This species belongs to the C. boninense species complex (Damm et al. 2012) and was previously reported causing anthracnose on strawberry leaves in Taiwan (Chung et al. 2020). To our knowledge, this is the first report of C. karstii causing anthracnose on strawberry in Brazil. The accurate identification of the pathogen will assist in the disease management and resistance breeding. usly reported causing anthracnose on strawberry leaves in Taiwan (Chung et al. 2020). To our knowledge, this is the first report of C. karstii causing anthracnose on strawberry in Brazil. The accurate identification of the pathogen will assist in the disease management and resistance breeding.
RESUMEN
The aim of this study was to evaluate the hygienic-sanitary quality of lamb meat sold in the city of Uruguaiana, Rio Grande do Sul (RS) by counting the indicator microorganisms and detecting pathogens such as Salmonella spp. and Listeria monocytogenes. Thirty-nine lamb meat samples were collected from 10 commercial establishments in Uruguaiana. The samples were subjected to counts of aerobic mesophilic microorganisms and enterobacteria, and to the detection of Salmonella spp. and L. monocytogenes, all following standard methods. The average counts of mesophilic microorganisms and enterobacteria were 6.08 log CFU/g (minimum 4.07 and max 6.87) and 4.73 log CFU/g (minimum 0 and max 5.88), respectively. For pathogens, L. monocytogenes was isolated from five samples (12.82%), with three samples in the same location. Only two samples (5.13%) were positive for Salmonella spp. The results demonstrated unsatisfactory hygienic-sanitary conditions because high counts of pathogens such as Salmonella spp. and L. monocytogenes. The counts of enterobacteria showed poor hygiene conditions during the various stages of production. The results also indicated fecal contamination, as Salmonella spp. and L. monocytogenes are present in the intestinal tract of both humans and animals. The high count of mesophilic microorganisms obtained could be owing to contaminated raw material or unsatisfactory processing, including unsanitary conditions and the inappropriate use of binomial time/temperature during storage.
O objetivo foi avaliar a qualidade higiênico-sanitária da carne de cordeiro comercializada na cidade de Uruguaiana, Rio Grande do Sul (RS) através da contagem dos microrganismos indicadores e detecção de patógenos como Salmonella spp. e Listeria monocytogenes. Trinta e nove amostras de carne de cordeiro foram coletadas em 10 estabelecimentos comerciais em Uruguaiana. As amostras foram submetidas a contagens de microrganismos aeróbios mesofilos e enterobactérias e à detecção de Salmonella spp. e L. monocytogenes, todas seguindo metodologias padrão. As contagens médias de microrganismos mesófilos e enterobactérias foram 6,08 log UFC/g (mínimo 4,07 e máximo 6,87) e 4,73 log UFC/g (mínimo 0 e máximo 5,88), respectivamente. Para os patógenos, L. monocytogenes foi isolado de cinco amostras (12,82%), com três amostras no mesmo local. Apenas duas (5,13%) foram positivas para Salmonella spp. Os resultados demonstraram condições higiênico-sanitárias insatisfatórias, baseado nas altas contagens de patógenos como Salmonella spp. e L. monocytogenes. As contagens de enterobactérias indicaram más condições de higiene durante as várias etapas de produção. Os resultados também indicaram contaminação fecal, uma vez que Salmonella spp. e L. monocytogenes estão presentes no trato intestinal de humanos e animais. A alta contagem de microrganismos mesófilos obtidos pode ser devida a matéria-prima contaminada ou processamento insatisfatório, incluindo condições insalubres e uso inadequado do binômio tempo/temperatura durante o armazenamento.
RESUMEN
Anxiety and depression are symptoms associated with ethanol withdrawal that lead individuals to relapse. In the kynurenine pathway, the enzyme indoleamine 2,3 dioxygenase (IDO) is responsible for the conversion of tryptophan to kynurenine, and dysregulation of this pathway has been associated with psychiatric disorders, such as anxiety and depression. The present study evaluated the early and late behavioral and biochemical effects of ethanol withdrawal in rats. Male Wistar rats were submitted to increasing concentrations of ethanol in drinking water during 21 days. In experiment 1, both control and withdrawal groups were submitted to a battery of behavioral tests 3, 5, 10, 19, and 21 days following ethanol removal. In experiment 2, animals were euthanized 3 days (short-term) or 21 days (long-term) after withdrawal, and the brains were dissected altogether, following kynurenine concentration analysis in prefrontal cortex, hippocampus, and striatum. Short-term ethanol withdrawal decreased the exploration of the open arms in the elevated plus-maze. In the forced swimming test, long-term ethanol-withdrawn rats displayed higher immobility time than control animals. Ethanol withdrawal altered neither locomotion nor motor coordination of rats. In experiment 2, kynurenine concentrations were increased in the prefrontal cortex after a long-term period of withdrawal. In conclusion, short-term ethanol withdrawal produced anxiety-like behaviors, while long-term withdrawal favored depressive-like behaviors. Long-term ethanol withdrawal elevated kynurenine levels, specifically in the prefrontal cortex, suggesting that the depressive-like responses observed after long-term withdrawal might be related to the increased IDO activity.
Asunto(s)
Encéfalo/enzimología , Etanol , Indolamina-Pirrol 2,3,-Dioxigenasa , Síndrome de Abstinencia a Sustancias , Animales , Ansiedad , Depresión , Indolamina-Pirrol 2,3,-Dioxigenasa/metabolismo , Quinurenina/análisis , Masculino , Ratas , Ratas WistarRESUMEN
ABSTRACT: This study evaluated the sanitary hygienic quality and the presence of pathogenic microorganisms in raw meats and fresh sausages marketed in the city of Uruguaiana, Rio Grande do Sul (RS), Brazil. We analyzed 238 samples of fresh sausages, beef, pork, and chicken from 18 commercial establishments (butchers, supermarkets, and groceries). Samples were subjected to enumerate hygiene indicator microorganisms (mesophilic aerobes and enterobacteria) and detection of Salmonella spp. and Listeria monocytogenes. The mean counts of mesophilic aerobes and enterobacteria were 5.09 and 3.54 log CFU/g, respectively. Beef samples presented the highest frequency of Salmonella spp. (7.93%) and fresh sausages present the highest frequency of L. monocytogenes (19.04%). Among the analyzed samples, 43.70% did not comply with the microbiological parameters established by the Brazilian Ministry of Health. The presence of Salmonella spp. and L. monocytogenes in different samples and commercial establishments demonstrate the failures of good manufacturing practices in industrial environmental and retails points and the need to train food handlers to reduce the exposure of consumers to potential risks.
RESUMO: Este estudo teve como objetivo avaliar a qualidade higiênica sanitária e a presença de microrganismos patogênicos em carnes in natura e linguiças frescais comercializadas na cidade de Uruguaiana, Rio Grande do Sul (RS), Brasil. Foram analisadas 238 amostras de linguiças, carne bovina, suína e de frango de 18 estabelecimentos comerciais (açougues, supermercados e mercearias). As amostras foram submetidas à enumeração de microrganismos indicadores de higiene (aeróbios mesófilos e enterobactérias) e detecção de Salmonella spp. e Listeria monocytogenes. As contagens médias de aeróbios mesófilos e enterobactérias foram 5,09 e 3,54 log UFC/g, respectivamente. Salmonella spp. esteve presente mais frequentemente em amostras de carne bovina (4,91 %), enquanto L. monocytogenes foi mais frequente em linguiças frescais (19,04 %). Das amostras analisadas, 43,70 % não atenderam aos parâmetros microbiológicos estabelecidos pelo Ministério da Saúde. A presença de Salmonella spp. e L. monocytogenes em diferentes amostras e estabelecimentos comerciais demonstra falhas de boas práticas de fabricação na indústria e pontos de venda, além da necessidade de treinar manipuladores de alimentos para reduzir a exposição dos consumidores à riscos.
RESUMEN
This study evaluated the sanitary hygienic quality and the presence of pathogenic microorganisms in raw meats and fresh sausages marketed in the city of Uruguaiana, Rio Grande do Sul (RS), Brazil. We analyzed 238 samples of fresh sausages, beef, pork, and chicken from 18 commercial establishments (butchers, supermarkets, and groceries). Samples were subjected to enumerate hygiene indicator microorganisms (mesophilic aerobes and enterobacteria) and detection of Salmonella spp. and Listeria monocytogenes. The mean counts of mesophilic aerobes and enterobacteria were 5.09 and 3.54 log CFU/g, respectively. Beef samples presented the highest frequency of Salmonella spp. (7.93%) and fresh sausages present the highest frequency of L. monocytogenes (19.04%). Among the analyzed samples, 43.70% did not comply with the microbiological parameters established by the Brazilian Ministry of Health. The presence of Salmonella spp. and L. monocytogenes in different samples and commercial establishments demonstrate the failures of good manufacturing practices in industrial environmental and retails points and the need to train food handlers to reduce the exposure of consumers to potential risks.(AU)
Este estudo teve como objetivo avaliar a qualidade higiênica sanitária e a presença de microrganismos patogênicos em carnes in natura e linguiças frescais comercializadas na cidade de Uruguaiana, Rio Grande do Sul (RS), Brasil. Foram analisadas 238 amostras de linguiças, carne bovina, suína e de frango de 18 estabelecimentos comerciais (açougues, supermercados e mercearias). As amostras foram submetidas à enumeração de microrganismos indicadores de higiene (aeróbios mesófilos e enterobactérias) e detecção de Salmonella spp. e Listeria monocytogenes. As contagens médias de aeróbios mesófilos e enterobactérias foram 5,09 e 3,54 log UFC/g, respectivamente. Salmonella spp. esteve presente mais frequentemente em amostras de carne bovina (4,91 %), enquanto L. monocytogenes foi mais frequente em linguiças frescais (19,04 %). Das amostras analisadas, 43,70 % não atenderam aos parâmetros microbiológicos estabelecidos pelo Ministério da Saúde. A presença de Salmonella spp. e L. monocytogenes em diferentes amostras e estabelecimentos comerciais demonstra falhas de boas práticas de fabricação na indústria e pontos de venda, além da necessidade de treinar manipuladores de alimentos para reduzir a exposição dos consumidores à riscos.(AU)
Asunto(s)
Análisis de los Alimentos/legislación & jurisprudencia , Higiene Alimentaria/legislación & jurisprudencia , Inspección de Alimentos/métodos , Salmonella , Listeria monocytogenesRESUMEN
Evaluating and characterizing production systems using farm characteristics allows the diagnosis of failing points. This diagnosis can be used to improve the productive and zootechnical indices. Little is known about the milk production systems in the state of Rio Grande do Sul, therefore, the aim of this study was to characterize the milk production systems of the Northwest, Center Western and Southwest mesoregions of Rio Grande do Sul, considering the infrastructure, milk handling, milk quantity and composition, and nutritional intake of the cattle. To conduct this study, 38 Milk Production Units (MPUs) registered at the Municipal Secretaries of Agriculture and Emater/Ascar-RS were randomly selected. After being randomly selected, the dairy farms were visited and a semi-structured guide questionnaire was applied and milk samples were collected from expansion tanks. The milk was analyzed for somatic cell counts (SCC) and total bacterial counts (TBC). Data were evaluated through principal component analysis and cluster analysis. Multivariate analysis allowed the investigated variables to be reduced into two main components (CP1 and CP2). These two showed eigenvalues greater than 1 (alpha> 1) and together explained 55.05% of the characteristics variability of the 38 MPUs studied. CP1 contemplated productive capacity and factors related to nutritional management of the MPUs, interfering directly with reproductive performance. CP2 comprised milk handling and daily production. Using these main variables, the data set generated from the 38 MPUs studied were adjusted and classified into five groups (G1, G2, G3, G4, and G5). The characteristics of these groups differed statistically especially in infrastructure and nutritional management of the cattle. Due to their particularities, each of these five groups of MPUs requires strategic technical interventions to improve their productive indexes. KEYWORDS: Management. Production Systems. Milk Quality. INTRODUCTIONThe milk activity is developed in milk production units (MPU) distributed throughout the Brazilian territory (WILLERS et al., 2014). These units have different characteristics regardless of the technological level adopted. The daily production and monthly remuneration play a fundamental role in the generation of jobs and income, moving the economy of the regions where they are inserted. However, the activity is developed in the MPUs under production systems with a wide range of organizational, technological, and production scales (BODENMÜLLER FILHO et al., 2010; WINCK; THALER NETO, 2012). Thus, each productive system has peculiar characteristics, which directly affect the productive efficiency of the MPUs. Rio Grande do Sul is the state in Brazil that produces the second largest volume of milk. In this Received: 08/04/19 Accepted: 30/12/19
Avaliar e caracterizar sistemas de produção utilizando as características das propriedades permite diagnosticar os pontos falhos, visando melhorar os índices produtivos e zootécnicos, desse modo pouco se sabe sobre os sistemas de produção de leite no estado do Rio Grande do Sul. Assim, objetivou-se com este estudo caracterizar os sistemas de produção de leite das mesorregiões Noroeste, Centro-Oeste e Sudoeste do Rio Grande do Sul, considerando a infra-estrutura, a quantidade, qualidade e composição do leite e o manejo nutricional dos rebanhos. Para conduzir este estudo, 38 Unidades de Produção de Leite (UPLs) registradas nas Secretarias Municipais de Agricultura e Emater/Ascar-RS foram selecionadas aleatoriamente. Depois, as fazendas leiteiras foram visitadas, um questionário semi-estruturado foi aplicado e amostras de leite do tanque foram coletadas. O leite foi analisado quanto à contagem de células somáticas (CCS) e à contagem bacteriana total (CBT). Os dados foram avaliados através de análise de componentes principais e análise de cluster. A análise multivariada permitiu que as variáveis investigadas fossem reduzidas em dois componentes principais (CP1 e CP2). Estes apresentaram autovalores maiores que 1 (alfa> 1) e juntos explicaram 55,05% da variabilidade das características das 38 UPLs estudadas. O CP1 contemplou a capacidade produtiva e os fatores relacionados ao manejo nutricional das UPLs, interferindo diretamente no desempenho produtivo. CP2 compreendeu manipulação de leite e produção diária. Utilizando essas variáveis principais, os dados gerados a partir das 38 UPLs estudadas foram ajustados e estas classificadas em cinco grupos (G1, G2, G3, G4 e G5). As características desses grupos diferiram estatisticamente, especialmente em infraestrutura e manejo nutricional do rebanho. Devido a suas particularidades, cada um desses cinco grupos de MPUs requer intervenções técnicas estratégicas para melhorar seus índices produtivos.
Asunto(s)
Sustitutos de la Leche HumanaRESUMEN
ABSTRACT: We aimed to perform serotyping and the antimicrobial resistance profile of Salmonella spp. and Listeria monocytogenes strains isolated from raw meats imported illegally into Brazil along the borders of Argentina and Uruguay. Distinct isolates of Salmonella spp. (n = 6) and L. monocytogenes (n = 25) obtained from 270 of these food products of earlier work were serotyped and tested for antimicrobial resistance by agar disk diffusion method. For strains that were considered phenotypically resistant, antimicrobial resistance genes were investigated: strA, strB, floR, tetA, tetB, blaZ, blaTEM, ermB, ermC, and ereB to Salmonella sp. and blaZ and mecA to L. monocytogenes. All Salmonella isolates were identified as Salmonella Infantis; they were multidrug resistant and harbored the genes blaTEM (n = 6), strA (n = 1), strB (n = 1), floR (n = 1), ermB (n = 1), tetA (n = 3), and tetB (n = 3). L. monocytogenes isolates belonged to serovars 1/2a (n = 1), 1/2b (n = 14), 1/2c (n = 2), and 4b (n = 8), showed resistance only to penicillin G (n = 12), and did not show the blaZ and mecA genes. The results demonstrated that illegal foods that are commercialized in the Brazilian international border with Argentina and Uruguay may harbor foodborne pathogens, and some of them have multidrug resistance characteristics, such as Salmonella, emphasizing the need for greater control of international food transit in Brazil, especially in the region evaluated.
Asunto(s)
Listeria monocytogenes , Antibacterianos/farmacología , Argentina , Brasil , Farmacorresistencia Bacteriana , Microbiología de Alimentos , Salmonella , UruguayRESUMEN
Entre los agentes patógenos causantes de enfermedades transmitidas por alimentos, Salmonella es considerado el más importante para la salud pública, y el serotipo S. Enteritidis los más involucrados en los casos de infección humana. Por lo tanto, esta revisión se centra en aspectos relacionados con la epidemiología de la salmonelosis y Salmonella, así como la importancia de pollo crudo como fuente de contaminación para las superficies de corte utilizadas en las cocinas domésticas para la manipulación de los alimentos y su posterior propagación a través de la contaminación cruzada, los alimentos listos para el consumo.
Among the pathogens responsible for causing food-borne diseases, Salmonella is considered the most important for public health, and the serotype S. Enteritidis is the most involved in cases of human infection. Thus, this literature review focuses on aspects related to the epidemiology of salmonellosis and Salmonella, as well as the importance of raw chicken as a source of contamination for cutting boards used in domestic kitchens for food handling and its subsequent spread through cross contamination to ready to eat foods.
Dentre os patógenos responsáveis por doenças transmitidas por alimentos, Salmonella é considerado o de maior importância para a saúde pública, sendo o sorotipo S. Enteritidis o mais frequentemente envolvido nos casos de infecção em humanos. Esta revisão aborda aspectos relacionados à epidemiologia da salmonelose e Salmonella, assim como a importância do frango cru como fonte de contaminação para superfícies de corte utilizadas nas cozinhas domésticas para manipulação de alimentos e sua consequente disseminação, através da contaminação cruzada, para alimentos prontos para o consumo.