Polymorphisms and haplotypes of TOLLIP and MUC5B are associated with susceptibility and survival in patients with fibrotic hypersensitivity pneumonitis.

INTRODUCTION AND OBJECTIVES: Hypersensitivity pneumonitis (HP) is an interstitial lung disease with diverse clinical features that can present a fibrotic phenotype similar to idiopathic pulmonary fibrosis (IPF) in genetically predisposed individuals. While several single nucleotide polymorphisms (SNPs) have been associated with IPF, the genetic factors contributing to fibrotic HP (fHP) remain poorly understood. This study investigated the association of MUC5B and TOLLIP variants with susceptibility, clinical presentation and survival in Portuguese patients with fHP. MATERIAL AND METHODS: A case-control study was undertaken with 97 fHP patients and 112 controls. Six SNPs residing in the MUC5B and TOLLIP genes and their haplotypes were analyzed. Associations with risk, survival, and clinical, radiographic, and pathological features of fHP were probed through comparisons among patients and controls. RESULTS: MUC5B rs35705950 and three neighboring TOLLIP variants (rs3750920, rs111521887, and rs5743894) were associated with increased susceptibility to fHP. Minor allele frequencies were greater among fHP patients than in controls (40.7% vs 12.1%, P<0.0001; 52.6% vs 40.2%, P = 0.011; 22.7% vs 13.4%, P = 0.013; and 23.2% vs 12.9%, P = 0.006, respectively). Haplotypes formed by these variants were also linked to fHP susceptibility. Moreover, carriers of a specific haplotype (G-T-G-C) had a significant decrease in survival (adjusted hazard ratio 6.92, 95% CI 1.73-27.64, P = 0.006). Additional associations were found between TOLLIP rs111521887 and rs5743894 variants and decreased lung function at baseline, and the MUC5B SNP and radiographic features, further highlighting the influence of genetic factors in fHP. CONCLUSION: These findings suggest that TOLLIP and MUC5B variants and haplotypes may serve as valuable tools for risk assessment and prognosis in fibrotic hypersensitivity pneumonitis, potentially contributing to its patient stratification, and offer insights into the genetic factors influencing the clinical course of the condition.

Chronic Aerobic Training at Different Volumes in the Modulation of Macrophage Function and in vivo Infection of BALB/c Mice by Leishmania major.

Physical inactivity is one of the main causes of chronic diseases; however, strenuous exercise can induce immunosuppression. Several studies suggest that moderate amounts of exercise lead to a Th1 response, favoring the resolution of infections caused by intracellular microorganisms, while high volumes of exercise tend to direct the response to Th2, favoring infection by them. Leishmaniasis is a parasitic disease promoted by parasites of the Leishmania genus, with clinical manifestations that vary according to the species of the parasite and the immune response of the host. The experimental Leishmania major-BALB/C mouse model provides a good model for the resistance (Th1 response) or susceptibility (Th2 response) that determines the progression of this infection. The aim of this study was to evaluate the effect of aerobic training at different volumes on modulation of in vitro macrophage infection by L. major, as well as to assess the effect of high volume (HV) aerobic training on the development of L. major in vivo in BALB/c mice. Uninfected animals were submitted to various exercise volumes: none (SED), light (LV), moderate (MV), high (HV), very high (VHV), and tapering (TAP). The macrophages of these animals were infected by L. major and the LV and MV groups showed a decrease in the infection factor, while the VHV showed an increase in the infection factor, when treated with LPS. The cytokine concentration pattern measured in the supernatants of these macrophages suggested a predominant Th1 response profile in the LV and MV groups, while the Th2 profile predominated in the VHV and TAP groups. Groups of BALB/C mice infected with L. major were subjected to high volume (iHV) or non-periodized high volume (iNPHV) exercise or kept sedentary (iSED). The exercised animals suffered a significant increase in injuries caused by the parasites. The animals in the group submitted to high volume exercise (iHV) showed visceralization of the infection. These data strongly suggest that a very high volume of aerobic training increased the susceptibility of BALB/C mice to L. major infection, while moderate distribution of training loads promoted immunological balance, better controlling the infection by this parasite.

Dental age assessment by the pulp/tooth area proportion in cone beam computed tomography: is medico-legal application for age estimation reliable?

INTRODUCTION: Forensic dentistry has, as one of its main goals, the identification of living and/or deceased individuals, based on the individual features of the teeth. One of the identification criteria to be established is the chronological age. Several authors, including Kvaal, have developed age estimation methods based on secondary dentine deposition. Nowadays, three-dimensional imaging tests, such as Cone Beam Computed Tomography (CBCT), are used in age estimation. OBJECTIVE: The aims of this research project were to validate Kvaal's method and its variables in age estimation and to create new linear regression formulae to better represent the study sample. METHODS: We selected 158 CBCT, with a total of 402 sound teeth (central incisors, lateral incisors and canines). The necessary measurements and ratios were calculated in both coronal and sagittal sections, with XelisDental . The formulae developed by Kvaal for age estimation calculation were applied. Subsequently, the results were statistically analyzed. RESULTS AND DISCUSSION: The intraclass correlation coefficients from the two measurements ranged from 0.918 to 0.997. The calculated age estimation had a mean error of -21.4years (coronal section) and -26.3years (sagittal section). The t test revealed statistically significant differences between chronological age and estimated age. The absolute values of Pearson's correlation coefficient between age and the two Kvaal variables ranged from 0.06 to 0.38 and from 0.06 to 0.55. The coefficients of determination are lower than in the original study (between 0.03 and 0.39). In the linear regression formulae, the coefficients of determination ranged from 0.07 to 0.41. CONCLUSION: This investigation concludes a non-reproducibility of Kvaal's method in the Portuguese population when applied in CBCT, with statistically significant differences between the chronological age and the dental age, estimated by the pulp/ tooth proportion method, based on the teeth analyzed in this study.

Rhinogenic and sinus headache - Literature review.

BACKGROUND: Headache is a common, yet challenging symptom to evaluate given its wide range of clinical presentations and different etiologies. For centuries, conceptual understanding of headache causation has been attributed to anatomic abnormalities of the nose and paranasal sinuses. METHODS: Structured literature review. RESULTS: The number of cases, categorized as migraines or other primary headaches, misdiagnosed as a "sinus headache" is high in the literature, ranging from 50 to 80%. The potential mechanisms for rhinogenic headaches were classically described as pain secondary to prolonged mucosal contact points, hypoxia in the paranasal sinuses secondary to poor ventilation, or pressure caused by the growth of nasal polyps. Additionally, other mechanisms were described and are still being studied. Corrective surgery for mucosal contact points in the nasal cavity is deemed necessary for relieving the headache, although patient outcomes are variable. CONCLUSION: Delay in proper diagnosis and treatment negatively impact patient quality of life. Most cases of "sinus headache" or "rhinogenic headache" seen in clinical practice are in fact misdiagnosed as either primary headaches or migraines. Because of increased misdiagnoses, Otolaryngologists should establish a direct and precise diagnosis congruent with a chief complaint being a headache. Vital information such as a good clinical history, well-performed nasal endoscopy, and occasional CT scan may decrease misdiagnosis probability.

Antibodies against Leptospira spp. in bovine serum samples from several Brazilian states analyzed in the period from 2007 to 2015 / [Anticorpos contra Leptospira spp. em amostra de soro bovino de diversos estados brasileiros analisados no período entre 2007 e 2015]

Bovine leptospirosis assumes great economic importance since it affects several production aspects. Therefore, knowledge about the occurrence and distribution of this disease is fundamental to adopt the correct prevention measures. The present study aimed to evaluate the frequency of anti-Leptospira spp. antibodies in 24,483 bovine serum samples received between 2007 to 2015 from 21 Brazilian states. Of these, 8,643 (35.3%) were reagents in the microscopic agglutination test to one or more serovars of Leptospira spp. The most frequent serovars were Wolffi (61.47%), Tarassovi (9.62%) and Pomona (7.20%). Hardjo serovar presented a prevalence of 6.27%. Among the 21 states analyzed, the State of Pernambuco had the highest frequency with 88.24% and the State of São Paulo was the origin of the largest number of analyzed samples (13,838), with a frequency of 31.54% of reagents. The results demonstrate a high exposure to several serovars of Leptospira spp. in bovine species in Brazilian states, showing the importance of adopting prophylactic measures in order to reduce the risk of infection in this specie.(AU)

Com o objetivo de avaliar a frequência de anticorpos anti-Leptospira spp., foram analisadas 24.483 amostras de soro sanguíneo bovino, provenientes de 21 estados brasileiros, recebidas no período de 2007 a 2015. Destas, 8.643 (35,3%) foram reagentes no teste de soroaglutinação microscópica a uma ou mais sorovariedades de Leptospira spp., e as sorovariedades com maior frequência foram Wolffi (61,47%), Tarassovi (9,62%) e Pomona (7,20%). A sorovariedade Hardjo apresentou prevalência de 6,27%. Entre os 21 estados analisados, o estado de Pernambuco apresentou a maior frequência, com 88,24%, e o estado de São Paulo foi a origem do maior número de amostras para análise, 13.838, com frequência de 31,54% de reagentes.(AU)

Revisiting the Phlebotominae subfamily records in Bahia, Brazil.

Brazil is one of the countries that concentrates 90% of all tegumentary and visceral leishmaniases cases and Bahia is one of the highly affected states. In the present report, we consolidated secondary data from several complementary databases that allowed us to record the sand fly species identified including areas of Leishmania spp. transmission in the state of Bahia. We then overlayed the geographical distribution data onto maps of vegetational aspects found across the state. Overall, 21 602 records of phlebotomine sand flies occurrence between 1949 and 2016 were analysed, encompassing 85% of Bahia's municipalities. Seventy-six sand fly species under 17 genera were enlisted. Among described species, 27 were proven or putative Leishmania spp. vectors and three were considered exclusively endemic in the state. Lutzomyia longipalpis, Nyssomyia intermedia and Nyssomyia whitmani were found in 74, 29 and 27% of municipalities, respectively. Salvador, the state capital and major city presented records for 21 different sand fly species, including known vectors for leishmaniasis. In particular, a wide distribution of Evandromyia sallesi was detected for this city. This consolidated account on phebotomine fauna and distribution may be explored for improving the planning and deployment of vector-focused leishmaniasis control measures in affected areas of Bahia.

Photophysical and Theoretical Interpretation of the Insensitive Emission to Temperature of a Metallopolymer Containing Europium Ions.

The luminescence and thermochromic properties of a europium-containing metallopolymer were investigated in experimental and theoretical aspects using the same polymer backbone complexed with two different contents of europium ions (25 and 65% molar). The polymer presented an emission insensitive to temperature variation which was attributed to a balance between two factors: the first is the "stiffening effect" on the polymer backbone brought about by ion complexation, and the second is the interconnection of the alkyl chains because of the rotation of the bipyridine sites required for the complexation.

Resposta hematológica do cascudo ornamental amazônico Peckoltia oligospila ao estresse de transporte / Hematological response of Amazonian ornamental Peckoltia oligospila subjected to stress by transportation

O objetivo deste trabalho foi avaliar as respostas hematológicas do acari-bola Peckoltia oligospila submetido ao estresse de transporte. Variações nos parâmetros de sangue foram analisadas às zero, seis, 24, 48, 72 e 96 horas após o transporte. Respostas ao estresse foram observadas entre zero e seis horas do transporte, mas a maioria dos parâmetros retornou aos valores basais em 24 horas. O tempo de zero hora (momento imediato após transporte) foi o mais crítico, com valores elevados de glicemia, eritrócitos e eritroblastos. Respostas secundárias tardias foram observadas para a proteína plasmática total, o volume corpuscular médio (VCM) e a hemoglobina corpuscular média (HCM) em seis horas após o transporte dos peixes, retornando aos valores basais após esse período. O número de leucócitos não sofreu alterações após o transporte. O estresse de transporte não comprometeu a fisiologia de P. oligospila, o que indica que esse peixe é resistente ao estresse se comparado com outras espécies. Porém, recomenda-se que não se realize qualquer outro procedimento estressante durante pelo menos 24 horas da recuperação dos peixes após transporte, para garantir a saúde e a sobrevivência dos animais transportados.(AU)

The objective of this work was to evaluate the hematological responses of bola-pleco (Peckoltia oligospila) undergoing the stress of transportation. Variations on blood parameters were analyzed at 0, 6, 24, 48, 72 and 96h after transportation. Responses to stress were detected from 0 to 6h after the transportation of fish, however, most parameters returned to baselines values within 24h of transportation. The moment of 0h was the most critical, presenting higher values of glycemia, erythroblasts and erythrocytes. Late secondary responses were observed to total plasmatic protein, mean corpuscular volume (MCV) and mean corpuscular hemoglobin (MCH) at 6h, returning to baselines values after this time. Leukocyte number was not affected by stress of transportation. The stress by transportation was not severe to influence the health of P. oligospila, indicating that fish is resistant to stress if compared to other species. However, we recommended no stressful procedures for at least 24 hours for recovery, in order to ensure health and survival of fish.(AU)

Impact of copper sulfate application at an urban Brazilian reservoir: A geostatistical and ecotoxicological approach.

A landscape ecotoxicology approach was used to assess the spatial distribution of copper in the recent bottom sediment (surficial sediment) of a Brazilian subtropical reservoir (the Guarapiranga reservoir) and its potential ecotoxicological impacts on the reservoir ecosystem and the local society. We discuss the policies and procedures that have been employed for the management of this reservoir over the past four decades. Spatial heterogeneity in the reservoir was evaluated by means of sampling design and statistical analysis based on kriging spatial interpolation. The sediment copper concentrations have been converted into qualitative categories in order to interpret the reservoir quality and the impacts of management policies. This conversion followed the Canadian Water Framework Directive (WFD) ecotoxicological concentration levels approach, employing sediment quality guidelines (SQGs). The SQG values were applied as the copper concentration thresholds for quantitative-qualitative conversion of data for the surficial sediment of the Guarapiranga. The SQGs used were as follows: a) interim sediment quality guideline (ISQG), b) probable effect level (PEL), and c) regional reference value (RRV). The quantitative results showed that the spatial distribution of copper in the recent bottom sediment reflected the reservoir's management policy and the copper application protocol, and that the copper concentrations varied considerably, ranging from virtually-zero to in excess of 3gcopper/kgds. The qualitative results demonstrated that the recent bottom sediment was predominantly in a bad or very bad condition, and could therefore have impacts on the local society and the ecosystem. It could be concluded that the management policy for this reservoir was mainly determined by the desire to minimize short-term costs, disregarding long-term socioeconomic and environmental consequences.

Brucella spp. in equines slaughtered in the south region of Brazil / Anticorpos contra Brucella spp. em equídeos da região Sul do Brasil abatidos em matadouro-frigorífico

Bacteria of the genus Brucella are widespread in many countries. These microorganisms can infect humans and many wild and domestic animal species. These bacteria have zoonotic potential, and can cause economic and public health problems since they can be transmitted by direct contact with sick animals, through consumption of contaminated milk, raw meat and its derivatives (Soares et al., 2015). Brucellosis is considered a chronic evolving disease, unusual in horses, predominantly caused by Brucella abortus. However, it is not characterized by reproductive disorders in horses, but primarily by abscess in the cervical region, bursa, tendons, and joints. Transmission is likely to occur via ingestion of contaminated water and pastures, especially in areas endemic for bovine brucellosis (Ribeiro et al., 2008). The slaughterhouse is a strategic point for obtaining information about the animal and animal products, edible or not. This study investigated the presence of anti-Brucella spp. immunoglobulins in the serum samples from horses slaughtered in a slaughterhouse in southern Brazil, to estimate the frequency of Brucella spp. antibodies and determine the spatial distribution of the cases.(AU)

Objetivou-se investigar a presença de imunoglobulinas anti-Brucella spp. em amostras de soros sanguíneos de equídeos abatidos em matadouro-frigorífico, sob Serviço de Inspeção Federal, localizado na região Sul do Brasil. Utilizaram-se 767 amostras de sangue de equídeos adultos abatidos no período de abril a maio de 2013. Os animais foram provenientes de 45 municípios dos estados do Rio Grande do Sul, Santa Catarina e Paraná. Para diagnóstico, foram utilizados os testes do antígeno acidificado tamponado (AAT), sendo os resultados positivos confirmados pelos testes de polarização fluorescente (TPF), reação de fixação de complemento (RFC) e 2-mercaptoetanol (2-ME). Foram sororreagentes no AAT 65 (8,47%) animais. Destes, apenas dois (3,07%) foram positivos também na RFC e três (4,62%) animais foram positivos no TPF. Apesar da baixa frequência de animais positivos para Brucella spp., pode-se afirmar que a infecção em equinos está presente na área estudada, o que é demonstrado pela presença de animais sororreatores. No âmbito da saúde animal, pública e ocupacional, sugere-se a atenção a essa doença, visando diminuir o risco de infecção.(AU)

Selection of prey to improve biological parameters of the predator Podisus nigrispinus (Dallas, 1851) (Hemiptera: Pentatomidae) in laboratory conditions / Seleção de presas para melhorar parâmetros biológicos do predador Podisus nigrispinus (Dallas, 1851) (Hemiptera: Pentatomidae) em condições de laboratório

Abstract Mass production of predatory stinkbugs in the laboratory is prioritized to release them into the field as part of IPM programs. Therefore, the aim of this study was to identify the best prey for rearing the predator Podisus nigrispinus (Dallas, 1851) (Hemiptera: Pentatomidae) among five different species of insect (three of Lepidoptera, one of Coleoptera, and one of Diptera). Second-instar P. nigrispinus nymphs were conditioned in transparent 1000-mL plastic pots, adults were placed in Petri dishes for mating, and both stages were maintained under controlled conditions (25 ± 1°C, 12 hours of photophase, 70 ± 10% RH). Nymphs and adults of P. nigrispinus consumed more Musca domestica (Linnaeus, 1758) (Diptera: Muscidae) larvae than the other tested prey. The consumption of fly larvae was 1.5 larvae/day/nymph and adults 1.7 larvae/day/adult. However, the number of eggs per female was less when the predator consumed M. domestica larvae (407.8 eggs/female) and most when consumed the Diatraea saccharalis (Fabricius, 1794) (Lepidoptera: Crambidae) larvae (797.7 eggs/female). Furthermore, the percentage of hatched eggs was greater when the predator females consumed D. saccharalis larvae (90.0%). D. saccharalis larvae is the best prey to rearing P. nigrispinus.

Resumo A produção massal de percevejos predadores no laboratório é priorizada para liberá-los no campo, como parte de programas de MIP. Portanto, o objetivo deste estudo foi identificar a melhor presa para a criação do predador Podisus nigrispinus (Dallas, 1851) (Hemiptera: Pentatomidae) entre cinco espécies diferentes de insetos (três de Lepidoptera, uma de Coleoptera e uma de Diptera). Ninfas de segundo ínstar de P. nigrispinus foram acondicionadas em recipientes plásticos transparentes de 1000 ml, adultos foram colocados em placas de Petri para o acasalamento e, ambos os estágios foram mantidos sob condições controladas (25 ± 1°C, fotofase de 12 horas e 70 ± 10% UR). Ninfas e adultos de P. nigrispinus consumiram mais larvas de Musca domestica (Linnaeus, 1758) (Diptera: Muscidae) do que as outras presas testadas. O consumo de larvas de mosca foi de 1,5 larvas/dia/ninfa e adultos 1,7 larvas/dia/adulto. No entanto, o número de ovos por fêmea foi menor quando o predador consumiu larvas de M. domestica (407,8 ovos/fêmea) e maior quando consumiu larvas de Diatraea saccharalis (Fabricius, 1794) (Lepidoptera: Crambidae) (797,7 ovos/fêmea). Além disso, a percentagem de eclosão de ninfas foi maior quando as fêmeas do predador consumiram larvas de D. saccharalis larvas (90,0%). Larvas de D. saccharalis é a melhor presa para a criação de P. nigrispinus.

Selection of prey to improve biological parameters of the predator Podisus nigrispinus (Dallas, 1851) (Hemiptera: Pentatomidae) in laboratory conditions.

Mass production of predatory stinkbugs in the laboratory is prioritized to release them into the field as part of IPM programs. Therefore, the aim of this study was to identify the best prey for rearing the predator Podisus nigrispinus (Dallas, 1851) (Hemiptera: Pentatomidae) among five different species of insect (three of Lepidoptera, one of Coleoptera, and one of Diptera). Second-instar P. nigrispinus nymphs were conditioned in transparent 1000-mL plastic pots, adults were placed in Petri dishes for mating, and both stages were maintained under controlled conditions (25 ± 1°C, 12 hours of photophase, 70 ± 10% RH). Nymphs and adults of P. nigrispinus consumed more Musca domestica (Linnaeus, 1758) (Diptera: Muscidae) larvae than the other tested prey. The consumption of fly larvae was 1.5 larvae/day/nymph and adults 1.7 larvae/day/adult. However, the number of eggs per female was less when the predator consumed M. domestica larvae (407.8 eggs/female) and most when consumed the Diatraea saccharalis (Fabricius, 1794) (Lepidoptera: Crambidae) larvae (797.7 eggs/female). Furthermore, the percentage of hatched eggs was greater when the predator females consumed D. saccharalis larvae (90.0%). D. saccharalis larvae is the best prey to rearing P. nigrispinus.

Avaliação do potencial biológico da Tabebuia aurea (Silva Manso) como fonte de moléculas bioativas para atividade antimicrobiana, antiedematogênica e antirradicalar / Biological potential assessment of Tabebuiaaurea (Silva Manso) as a source of bioactive molecules for antimicrobial, antiedematogenic and antiradical activity

RESUMO As espécies da família Bignoniaceae e do gênero Tabebuia são amplamente utilizadas na medicina tradicional e possuem um forte potencial terapêutico. Diante disso, objetivou-se avaliar o potencial biológico da Tabebuia aurea, determinando a atividade antimicrobiana; por meio do método da microdiluição em caldo, para a determinação da Concentração Inibitória Mínima (CIM); antiedematogênica, pelo ensaio de edema de orelha induzido por capsaicina; e antirradicalar, frente ao radical DPPH. Os extratos etanólicos de T. aurea não evidenciaram citotoxicidade, exceto o extrato etanólico da flor nas concentrações > 0,5 mg mL-1. O extrato etanólico da flor foi ativo com ação bactericida frente a S. epidermidis (CIM de 0,06 mg mL-1) enquanto o extrato etanólico da folha foi moderadamente ativo frente a S. epidermidis (CIM: 0,25 mg mL-1) e S. aureus (CIM: 0,50 mg mL-1) sugerindo ação bacteriostática para ambas as linhagens. Os dois extratos apresentaram ação antiedematogênica, com inibição do edema de 40,50% pelo extrato etanólico da flor e de 41,73% pelo extrato da folha. T. aurea não apresentou atividade antirradicalar. Os resultados comprovam o perfil antibacteriano e antiedematogênico com ausência de citotoxidade pela T. aurea. Sugere-se a continuação dos testes com frações e substâncias isoladas das flores e folhas da referida espécie vegetal, bem como de experimentos in vivo, como forma de agregar evidências visando à busca de novos fitoterápicos.

ABSTRACT The species of Bignoniaceae family and genus Tabebuia are widely used in the traditional medicine and have a great therapeutic potential. The aim of the current research was to evaluate the biological potential of the Tabebuia aurea, determining its antimicrobial activity by the microdilution broth method, to predict the anti-edematogenic Minimum Inhibitory Concentration (MIC) by ear edema assay induced by capsaicin; and the antiradical one, towards DPPH. The ethanol extracts of T. aureashowed no cytotoxicity, except for the flower ethanol extract in concentrations above > 0.5 mg mL-1. The ethanol extract of the flower was active, with bactericidal action, against S. epidermidis (MIC 0.06 mg ml-1) and the ethanol extract of moderately active recto S. epidermidis (CIM: 0.25 mg mL-1) and S. aureus (MIC: 0.50 mg mL-1) were bacteriostatic for both strains. Both extracts had antiedematogenic action on the inhibition of edema of 40.50% by the ethanol extract of the flower and 41.73% by leaf extract. T. aurea did not show antiradical activity. The results indicate the antibacterial and antiedematogenic profile with no cytotoxicity by the T. aurea. It suggests the continuation of tests with isolated fractions and substances of flowers and leaves of that plant species as well as in vivo trials, in order to enhance the evidences targeted on finding new herbal medicines.

Estudo do potencial antimicrobiano e citotóxico da espécie Pouteria venosa (Sapotaceae) / Study of antimicrobial and cytotoxic potential of Pouteria venosa species (Sapotaceae)

RESUMO Estudo experimental in vitro que objetivou investigar o potencial antimicrobiano e citotóxico de quatro frações e um extrato etanólico da espécie Pouteria venosa usada como planta medicinal. A atividade antimicrobiana foi determinada pelos testes de sensibilidade microbiana, como o método de difusão em disco e o método da microdiluição em caldo, para a determinação da Concentração Inibitória Mínima (CIM). Obteve-se a avaliação da citotoxicidade por meio do método colorimétrico do Metiltetrazolium. No estudo da atividade antimicrobiana, os principais resultados foram obtidos contra Staphylococus aureus para a fração AcOEt das cascas do caule, CIM de 125 µg/mL; Streptococcus pneumoniae e Proteus mirabilis para a fração AcOEt das cascas do caule, CIMde250 µg/mL; Staphylococus epidermidis, Pseudomonas aeruginosa para a fração AcOEt das folhas e cascas do caule, CIM de 250 µg/mL. Todas as amostras foram inativas para os fungos testados. A fração AcOEt das cascas do caule foi considerada atóxica, podendo ser utilizada em testes pré-clínicos in vivo

ABSTRACT Study of antimicrobial and cytotoxic potential of Pouteria venosa species (Sapotaceae). This experimental in vitro study aimed to investigate the antimicrobial and cytotoxic potential of four fractions and one ethanolic extract of the specie Pouteria venosa used as a medicinal plant. The antimicrobial activity was determined by microbial sensitivity tests, as the method of disk diffusion and the broth microdilution method for determination of Minimum Inhibitory Concentration (MIC). The evaluation of the cytotoxicity was obtained by the Metiltetrazolium colorimetric method. In the antimicrobial activity research, the main results were obtained against the Staphylococcus aureus for the AcOEt fraction of the stem bark MIC 125 µg/mL, Streptococcus pneumoniae and Proteus mirabilis for the AcOEt fraction from the stem bark, CIM 250 µg/mL, Staphylococcus epidermidis, Pseudomonas aeruginosa to the AcOEt fraction of the leaves and stem bark, MIC 250 µg/mL. All samples did not react for the fungi tested. The AcOEt fraction of the stem bark was considered non-toxic and can be used at in vivo pre-clinical testing

First Report of Ilyonectria macrodidyma Associated with Black Foot Disease of Grapevine in Brazil.

Cultivated grapevine (Vitis labrusca and V. vinifera) is of considerable economic importance to the Brazilian fruit industry for both fresh market consumption and for the production of wines, sparkling beverages, and juices. Black foot disease is caused by fungi of the genera Ilyonectria P. Chaverri & C. Salgado (anamorph: Cylindrocarpon Wollew.), Campylocarpon Halleen, Schroers & Crous, and Cylindrocladiella Boesew. In 2012, 4- to 40-year-old grapevines (Vitis spp.) showing reduced vigor, vascular lesions, necrotic root lesions, delayed budding, vine decline, and death were collected from seven locations at Rio Grande do Sul state, Brazil. Fungal isolations were made from root fragments and crown lesions (at least 2 cm above the bottom) on potato dextrose agar (PDA) medium added with 0.5 g L-1 streptomycin sulfate. Eight isolates were obtained and identified on the basis of morphological features and multi-gene analysis (rDNA-ITS, ß-tubulin, and histone H3) as Ilyonectria macrodidyma (Halleen, Schroers & Crous) P. Chaverri & C. Salgado. One representative isolate (Cy5UFSM) was used for more detailed morphological and molecular characterization, and pathogenicity confirmation. When incubated in the dark at 20°C for 7 to 10 days, colonies of felty straw-colored mycelium (3) 4.79 cm diameter on average were observed. No sporodochia or other fruiting bodies were produced on carnation leaf agar (CLA) medium after 30 days. Microconidia that were produced after 5 weeks on spezieller nährstoffarmer agar (SNA) medium with addition of two pieces of 1 cm2 filter paper showed ovoid and ellipsoid shape (6.4 × 3.6 µm) and one-septate macroconidia (17.3 × 4.1 µm). To confirm the species, primer pairs ITS1 and ITS4 (4); Bt2a and Bt2b; and H3-1a and H3-1b (2) were used to amplify the ITS1-5.8S rRNA-ITS2, part of the ß-tubulin and histone H3 genes, respectively. Sequences of these three regions showed 99, 100, and 100% of homology with I. macrodidyma, respectively. To confirm pathogenicity, 4-month-old rooted cuttings of V. labrusca cv. Bordô were inoculated by immersing them in a conidial suspension of the isolate (106 conidia ml-1) for 60 min (1). Thirty days later, inoculation was performed again by drenching the crown with 40 ml of 106 conidia ml-1 suspension to ensure infection of the roots. In the control treatment, plants were inoculated with sterile distilled water. Plants inoculated with I. macrodidyma showed necrosis of the leaf ribs, reduction in root mass, root and crown necrosis, browning of vessels, drying of shoots, and death. I. macrodidyma was re-isolated from the crown necrosis and vascular lesions, confirming Koch's postulates. To our knowledge, this is the first report of I. macrodidyma associated with black foot disease of grapevine in Brazil, which poses considerable threat to the industry unless management options are realized. References: (1) A. Cabral et al. Phytopathol. Mediterr. 51:340, 2012. (2) N. L. Glass et al. Appl. Environ. Microbiol. 61:1323, 1995. (3) R. W. Rayner. A Mycological Colour Chart. Commonwealth Mycological Institute and British Mycological Society, 1970. (4) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, CA, 1990.

First Report of Ilyonectria robusta Associated with Black Foot Disease of Grapevine in Southern Brazil.

In August 2012, symptoms of black foot disease were observed on 21-year-old grapevines (Vitis labrusca cv. Bordô; own-rooted cultivar) at Nova Pádua city, Rio Grande do Sul state, Brazil. Symptomatic plants showed reduced vigor, vascular lesions, decline and death of vines, and necrotic lesions on roots. Isolation of fungi associated with necrotic root tissue was made on potato dextrose agar (PDA) medium containing 0.5 g L-1 streptomycin sulfate. Cultures were incubated at 25°C for 7 days in darkness, and single-spore cultures were obtained from the colonies emerging from the diseased tissue. For morphological characterization, cultures were transferred to PDA and spezieller nährstoffarmer agar (SNA) medium with addition of two pieces of 1 cm2 filter paper. One representative isolate (Cy9UFSM) was used for morphological and molecular characterization and pathogenicity confirmation. After 10 days growth on PDA at 20°C in the dark, colonies were umber to chestnut in color (3), appeared cottony to felty in texture, and sporulated profusely. After 5 weeks on SNA and under dark conditions at 20°C, cultures formed macroconidia predominantly on simple conidiophores, 1 to 3 septate, with both ends slightly rounded. Macroconidia varied in size depending on the number of cells as follows: one-septate (23-) 27.7 (-31) × (4.5-) 5.8 (-7) µm; two-septate (26-) 30.1 (-34) × (5-) 5.6 (-6) µm; and three-septate (24-) 31.2 (-35) × (5-) 5.8 (-6.5) µm. Microconidia were observed and did not have a visible hilum (6-) 11.2 (-17) × (3.5-) 4.2 (-5) µm (n = 30 observations per structure). Brown, thick-walled globose to subglobose chlamydospores were produced abundantly on PDA, (8.5-) 13.8 (-17) µm. To confirm the species, primer pairs H3-1a and H3-1b (2) were used to amplify a portion the histone H3 gene. Sequence of this region showed 98% similarity with a reference sequence for Ilyonectria robusta (A.A. Hildebr.) A. Cabral & Crous (GenBank Accession No. JF735530). Thus, both morphological and molecular criteria supported identification of the strain as I. robusta. This isolate was deposited in GenBank as accession KF633172. To confirm pathogenicity, 4-month-old rooted cuttings of Vitis labrusca cv. Bordô were inoculated by immersing roots in a conidial suspension (106 ml-1) for 60 min. After inoculation, the cuttings were planted in 1-L bags containing commercial substrate (MecPlant). Thirty days later, each plant was re-inoculated by applying 40 ml of a conidial suspension (106 ml-1) to the commercial substrate. Ten single-vine replicates were used for each isolate, and 10 water-inoculated vines were included as controls. After 4 months, the inoculated plants showed a 22.5% reduction of root mass, with root and crown necrosis, browning of vessels, and 20% mortality. Control plants treated with water remained symptomless. The fungus was re-isolated from blackened tissue of wood from the basal end of rooted cuttings, thereby satisfying Koch's postulates. I. robusta was first associated with black foot disease of grapevine in Portugal in 2012 (1). To our knowledge, this is the first report in southern Brazil of I. robusta associated with black foot disease of grapevine. References: (1) A. Cabral et al. Mycol. Prog. 11:655, 2012. (2) N. L. Glass et al. Appl. Environ. Microbiol. 61:1323, 1995. (3) R. W. Rayner. A mycological colour chart. Commonwealth Mycological Institute and British Mycological Society, 1970.

First Report of Fusarium equiseti Associated on Pecan (Carya illinoinensis) Seeds in Brazil.

Pecan [Carya illinoinensis (Wangenh.) K. Koch] is an important producing nut tree that has been intensively cultivated in the state of Rio Grande do Sul (Brazil) in recent decades. This species is commonly grown in association with other crops and more often with cattle or sheep. An elevated incidence of the fungal genus Fusarium was observed during a quality control seed assay of pecan seeds obtained from orchards in the city of Anta Gorda (28°53'54.7″ S, 52°01'59.9″ W). Concomitantly, seedlings of this species, cultivated in a nursery, showed foliar necrosis, wilt, and root rot. The fungus was thereafter isolated from the seeds (from original seeds lots) and subcultured from single spores. Cultures were purified in order to perform pathogenicity tests. The isolated Fusarium sp. was increased on autoclaved wet corn kernels that were incubated for 14 days (1), and then were mixed with commercial substrate (sphagnum turf, expanded vermiculite, dolomitic limestone, gypsum, and NPK fertilizer) in plastic trays (capacity 7 L), with drainage holes. Twenty seeds were sowed and 90 days later, evaluations were undertaken. Forty percent of the seedlings presented symptoms, i.e., foliar necrosis and wilt owing to root rot. Fusarium sp. was re-isolated from the affected roots by transferring hyphal tips to potato dextrose agar (PDA) and carnation leaf agar (CLA) medium in petri dishes in order to identify the species morphologically. On PDA, the colony pigmentation was yellowish brown and the aerial mycelium was whitish to peach; macroconidia were relatively long and narrow (31.75 × 4.02 µm), with 5 septa on average, and whip-like bent apical cells (2). Chlamydospores were not observed on PDA or CLA. Primer pairs ITS1 and ITS4 (3) and EF1-T and EF1-1567R (4) were employed to amplify the internal transcribed spacer (ITS) and elongation factor-1α (TEF 1-α) regions, respectively. The resulting DNA sequences showed 99% for ITS and 98% for TEF 1-α similarity with Fusarium equiseti (Corda) Sacc. and phylogenetic analysis grouped it with sequences of this species. The consensus sequence was submitted to GenBank and received the accession numbers KC810063 (ITS) and KF601580 (TEF 1-α). The pathogen was re-isolated on PDA and CLA substrate in order to complete Koch's postulates. The pathogenicity test was repeated with the same conditions described before and the results were confirmed. No symptoms were observed on the control seedlings. This species is considered a weak parasite (2); however, it has been reported causing wilt in Coffea arabica in Brazil (5). This pathogen could cause serious damage and high losses to seedling in commercial nurseries. Besides that, it could also carry the disease to the field causing further damage on established plants. To our knowledge, this is the first to report of F. equiseti causing foliar necrosis and wilt on C. illinoinensis in Brazil. References: (1) L. H. Klingelfuss et al. Fitopatol. Brasil. 32:1, 2007. (2) W. Gerlach and H. Nirenberg. The Genus Fusarium - a Pictorial Atlas. Biologische Bundesanstalt für Land- und Forstwirtschaft, Braunschweig, Germany, 1982. (3) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications, Academic Press, San Diego, CA, 1990. (4) S. A. Rehner and E. A. Buckley. Mycologia 97:84, 2005. (5) L. H. Pfenning and M. F. Martins. Page 283 in: Simpósio de Pesquisa dos Cafés do Brasil, 2000.

First Report of "Cylindrocarpon" pauciseptatum Associated with Black Foot Disease of Grapevine in Brazil.

Since 1999, the decline of American grapevines (Vitis labrusca L.) has been common in Rio Grande do Sul, Brazil (1). In August 2012, V. labrusca with black foot symptoms were collected in vineyards in the Serra Gaúcha Region. Symptomatic plants had low vigor, vascular lesions, delayed budding, and decline and death of vines. Symptomatic roots had necrotic lesions and reduced biomass. Fungal isolations were made from necrotic root and crown fragments (own-rooted cultivar) on potato dextrose agar (PDA) medium amended with 0.5 g L-1 streptomycin sulfate. Putative colonies of "Cylindrocarpon" pauciseptatum Schroers & Crous were obtained from single macroconidia isolations. Two isolates were used to confirm the identity of isolated colonies: Cy12UFSM and Cy13UFSM. After incubation in the dark for 10 days at 20°C, the isolated mycelial colonies, which were cottony white to felty in texture, became dark orange to brown. Both isolates produced chlamydospores in chains at 40 days. Chlamydospores of Cy12UFSM and Cy13UFSM were 9 to 12 µm and 5 to 11.5 µm in diameter. Sporodochia formation on carnation leaf agar (CLA) medium was observed after 30 days. To encourage development of conidia, the isolates were grown on spezieller nährstoffarmer agar (SNA) medium for five weeks at 20°C with addition of two pieces of 1 cm2 filter paper. Microconidia of Cy12UFSM were 4 to 8.5 × 3.5 to 5 µm and those of Cy13UFSM were 3.5 to 7.5 × 3 to 5 µm. Macroconida were predominantly 3-septate (Cy12UFSM was 36 to 45 × 7.5 to 9 µm and Cy13UFSM was 30 to 38 × 7.5 to 8 µm), but 1-, 2- septate macroconidia were observed. The sizes of the three spore types and colony morphology for our isolates were similar to those described by Schroers et al. (3) for "C." pauciseptatum. To further confirm the identity of Cy12UFSM and Cy13UFSM, multi-gene DNA sequence analysis (rDNA-ITS, ß-tubulin, and histone H3) was conducted using primer pairs ITS1 and ITS4 (4), Bt2a and Bt2b, and H3-1a and H3-1b (2), which amplify the ITS1-5.8S rRNA-ITS2 genes, part of the ß-tubulin gene, and the histone H3 gene, respectively. Sequences of these three regions had 99, 99, and 97% similarity with references sequences of "C." pauciseptatum (isolate Cy238; accessions ITS [JF735307]; ß-tubulin [JF735435], and histone H3 [JF735582], respectively). To evaluate pathogenicity, 4-month-old rooted cuttings of V. labrusca cv. Bordô were inoculated with two isolates by immersing them in a conidial suspension (106 conidia ml-1) for 60 min. Ten single-vine replicates were used for each isolate, and 10 water-inoculated vines were included as controls. Thirty days after inoculation, vines were re-inoculated with 40 ml of a 106 conidia ml-1 suspension to ensure root infection. After 4 months, the inoculated plants had reduced root mass relative to controls (39.18% for Cy12UFSM and 18.27% for Cy13UFSM). Inoculated plants also had root and crown necrosis, vascular lesions, shoot decline, and vine mortality (60 and 80% mortality for Cy12UFSM and Cy13UFSM, respectively). All water-inoculated control plants remained symptomless. The fungi Cy12UFSM and Cy13UFSM were re-isolated from infected woody tissues, confirming Koch's postulates. To our knowledge, this is the first report of "C." pauciseptatum associated with black foot disease of grapevine in Brazil, which may potentially impact the sustainability of grapevine nurseries and vineyard productivity. References: (1) L. R. Garrido et al. Fitopatol. Brasil. 29:548, 2004. (2) N. L. Glass et al. Appl. Environ. Microbiol. 61:1323, 1995. (3) H. J. Schoers et al. Mycol. Res. 112:82, 2008. (4) T. J. White et al. Amplification Pages 315-322 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, CA, 1990.