Effect of entomopathogenic fungi on the control of the coffee berry borer in the laboratory

Among the management and control tactics of the coffee berry borer Hypothenemus hampei (Ferrari), there is the use of entomopathogenic fungi. Due to the importance of prospecting isolates of entomopathogenic fungi for the control of the coffee berry borer, the objective of this study was to evaluate the efficiency of 26 isolates of entomopathogenic fungi in the control of this insect pest in the laboratory. The coffee berry borers were immersed in a solution adjusted to the concentration of 1 to 3 × 108 conidia/mL of each isolate and the control treatment (sterilized water). After seven days total mortality and confirmed mortality were evaluated. The isolates that caused the highest mortality and two commercial isolates were selected for evaluation of lethal concentration (LC50 and LC90) and lethal time (LT50 and LT90). Coffee berry borers were treated at different conidia concentrations for lethal concentration to assess total and confirmed mortality. For a lethal time, the coffee berry borers were treated at the concentration of 108 conidia/mL of the selected isolates, and, after two days and every 24 hours until the eighth day, the number of dead individuals was verified. Among the 26 isolates evaluated, 24 presented mortality higher than the control treatment, and three presented mortality higher than 85%. In the LC50 and LC90 assays, the IBCB 353 and IBCB 364 isolates were more lethal to H. hampei. In the LT50 and LT90 assays, the IBCB 66 and IBCB 353 isolates caused lethality in a shorter time.

Anti-hyperglycemic fraction from Alternanthera sessilis L. leaves gets elucidated following bioassay-guided isolation and mass spectrometry

Abstract The anecdotal use of Alternanthera sessilis L. as a relief for diabetes has been known in the Philippines for generations, and antidiabetic activity of similar varieties in other countries is likewise documented. However, the compounds responsible for this activity remain unclear. This study aims to isolate the anti-hyperglycemic fraction of local A. sessilis leaves and identify the compounds in this fraction. Methanol extract of A. sessilis leaves and its hexane, ethyl acetate (ASE), and water fractions were administered to alloxan-induced diabetic mice. ASE (250mg/kg) had the highest anti-hyperglycemic activity at 6-h post-treatment (25.81%±12.72%), with almost similar blood glucose reduction rate as metformin (30.13±3.75%, p=0.767). Repeated fractionation employing chromatographic separation techniques followed by in vivo anti-hyperglycemic assay yielded partially purified subfractions. A. sessilis ethyl acetate subfraction 4-2 (100mg/kg) displayed remarkable suppression of blood glucose rise in diabetic mice at 6-h post-treatment (26.45±3.75%, p<0.0001), with comparable activity with metformin (100mg/kg, 27.87±5.65%, p=0.652). Liquid chromatography/mass spectrometry showed eight distinct peaks, with four peaks annotated via the Traditional Chinese Medicine library and custom library for A. sessilis. Among these, luteolin, apigenin, ononin, and sophorabioside were identified as putative compounds responsible for the anti-hyperglycemic activity. This result provided basis for the reported anecdotal claims and potential utility of the local variety of A. sessilis leaves as sources of anti-hyperglycemic agents

Bacillus thuringiensis A21, un aislado no tóxico de elevada actividad larvicida residual contra Aedes aegypti / Bacillus thuringiensis A21, a non-toxic isolate of high residual larvicidal activity against Aedes aegypti

Introducción: Los productos derivados de Bacillus thuringiensis se han empleado con éxito en el control de insectos. Sin embargo, la poca residualidad promueve la búsqueda de alternativas. En Cuba diferentes estudios informan la evaluación y selección de aislamientos de B. thuringiensis con actividad larvicida contra Aedes aegypti: el aislado A21 se destaca por su elevada actividad larvicida y ausencia de betaexotoxina. Objetivo: Evaluar la toxicidad/patogenicidad aguda oral y actividad larvicida residual del aislado A21. Métodos: La evaluación de la toxicidad/patogenicidad aguda oral del aislado A21 se estableció en el modelo biológico Rattus norvegicus. Se realizaron observaciones clínicas diarias de los animales y se evaluó el peso corporal. Se estimó la eliminación y la infectividad de B. thuringiensis mediante análisis de muestras de heces, y de fluidos y órganos, respectivamente. Para determinar la actividad larvicida residual del aislado A21 frente a larvas de Ae. aegypti se realizaron bioensayos con diferentes volúmenes de recambios de agua semanal (total, parcial, no recambio) en los recipientes. La mortalidad obtenida se calculó a las 24-72 h. Resultados: Con el aislado A21 no se detectó toxicidad/patogenicidad aguda oral en Rattus norvegicus. En el estudio de residualidad, la mortalidad larvaria a las 72 h se mantuvo elevada (80-100 por ciento) hasta la semana 24. A partir de la semana 25 la mortalidad larvaria disminuyó (p < 0,05). Conclusiones: Se evidencia la baja toxicidad y la elevada actividad larvicida residual del aislado A21 contra Ae. aegypti y lo convierten en un candidato promisorio para el desarrollo de biolarvicidas. Estos productos biológicos podrían contribuir a mejorar las estrategias de control del vector existentes en Cuba(AU)

ABSTRACT Introduction: Bacillus thuringiensis-based products have been successfully used for insect control. However, their low residuality promotes the search for alternatives. In Cuba, different studies have informed about the evaluation and selection of B. thuringiensis isolates with larvicidal activity against Aedes aegypti: isolate A21 highlights for its high larvicidal activity and absence of beta-exotoxins. Objective: To evaluate the acute oral toxicity/pathogenicity and the residual larvicidal activity of isolate A21. Methods: The evaluation of the acute oral toxicity/pathogenicity of isolate A21 was established in the animal model Rattus norvegicus. Daily clinical observations of the animals were carried out, and their body weight was evaluated. The elimination and infectivity of B. thuringiensis were estimated by analyzing feces, and fluids and organs samples, respectively. To determine the residual larvicidal activity of isolate A21 to Ae. aegypti larvae, bioassays were conducted with different volumes of weekly water changes (total, partial, no change) in the containers. Mortality was calculated at 24h-72h. Results: No acute oral toxicity/pathogenicity was detected with isolate A21 in Rattus norvegicus. In the study of residuality, at 72h larval mortality remained high (80 percent-100%) until week 24, decreasing from week 25 (p < 0.05). Conclusions: It is evidenced the low toxicity and high residual larvicidal activity of isolate A21 against Ae. aegypti, which makes it a promising candidate for the development of biolarvicides. These biological products could contribute to the improvements of the existing vector control strategies in Cuba(AU)

Type C botulism in dogs from rural properties located in Goiânia, Brazil

Botulism is a disease usually fatal, caused by the ingestion of neurotoxins produced by Clostridium botulinum. In dogs, intoxication is caused by the ingestion of botulinum toxin type C, and animals often recover spontaneously. The present study describes the occurrence of type C botulism in two dogs domiciled on neighboring rural properties in the municipality of Goiânia, state of Goiás, Brazil, probably associated with ingestion of decomposing bovine carcass. Upon clinical evaluation, the dogs were alert in the lateral decubitus position with ascending flaccid paralysis, absence of eyelid reflexes, and reduced muscle tone. Due to their worsening clinical symptoms, the animals died within 12 h and 3 days after supportive treatment. Botulinum toxin type C was identified, in the serum and feces of both dogs, by seroneutralization in mice with homologous monovalent antitoxin. The results of the high-throughput gene sequencing showed that the abundance of C. botulinum in the fecal microbiota of one of the affected dogs was low (0.53%). In this way, the present study highlights the need of sanitary practices related to the appropriate collection and disposal of bovine carcasses in rural areas since they represent a risk factor for the occurrence of botulism in dogs domiciled on rural properties.

A comparative study on the virulence of entomopathogenic fungi against Trogoderma granarium (Everts) (Coleoptera: Dermestidae) in stored grains rice / Um estudo comparativo da virulência de fungos entomopatogênicos contra Trogoderma granarium (Everts) (Coleoptera: Dermestidae) em grãos armazenados de arroz

Entomopathogenic fungi (EPF) now a possible safer microbial control measure that could be considered as a substitute for chemical control of insect pests. Three EPF viz., Metarihizium anisopliae, Isaria furnosoroseus and Beauveria bassiana were evaluated for their virulence against the grubs of Khapra beetle, Trogoderma granarium (Everts) under laboratory conditions. The isolates were applied by two methods viz., diet incorporation and an immersion method with 3rd instar 20 grubs of T. granarium for each. The virulence of EPF was determined using percent mortality. Significantly higher mortality was observed in M. anisopliae applied through immersion (98.33%) and diet incorporation (93.33%) methods followed by B. bassiana (90.83 and 85.83%, respectively). The mortality caused by I. furnosoroseus was statistically lower in immersion and diet incorporation methods i.e. 81.67 and 73.33%, respectively. Based on the immersion method, all EPF were studied for multiple conidial concentration i.e., 1×104 , 1×105, 1×106, 1×107 and 1×108 under the same in-vitro conditions. All the isolates were pathogenic to grub of T. granarium at the highest conidial concentration. M. anisopliae was proved the most effective virulent resulting in 98.33% mortality of the pest with LT50 4.61 days at 1 × 108 conidial concentration followed by 90.83 and 81.67 percent mortality with 5.07 and 8.01 days LT50, in the application of B. bassiana and I. furnosoroseus, respectively. M. anisopliae showed higher efficacy and could be considered as promising EPF for the development of myco-insecticides against effective biocontrol of T. granarium.

Os fungos entomopatogênicos (FPE) são agora a possível medida de controle microbiano mais segura, que pode ser considerada um substituto para o controle químico de pragas de insetos. Três EPF viz., Metarihizium anisopliae, Isaria furnosoroseus e Beauveria bassiana foram avaliados quanto à sua virulência contra as larvas do besouro Khapra, Trogoderma granarium (Everts) em condições de laboratório. Os isolados foram aplicados por dois métodos, a saber: incorporação de dieta e um método de imersão com 20 larvas de T. granarium de 3º ínstar para cada um. A virulência do EPF foi determinada usando a mortalidade percentual. Mortalidade significativamente maior foi observada em M. anisopliae aplicado pelos métodos de imersão (98,33%) e incorporação de dieta (93,33%), seguido por B. bassiana (90,83% e 85,83%, respectivamente). A mortalidade causada por I. furnosoroseus foi estatisticamente menor nos métodos de imersão e incorporação de dieta, ou seja, 81,67% e 73,33%, respectivamente. Com base no método de imersão, todos os EPFs foram estudados para múltiplas concentrações de conídios, ou seja, 1 × 104, 1 × 105, 1 × 106, 1 × 107 e 1 × 108 nas mesmas condições in vitro. Todos os isolados foram patogênicos à larva de T. granarium na maior concentração de conídios. M. anisopliae provou ser o virulento mais eficaz, resultando em 98,33% de mortalidade da praga com LT50 4,61 dias na concentração de 1 × 108 conídios seguido por 90,83% e 81,67% de mortalidade com 5,07 e 8,01 dias LT50, na aplicação de B. bassiana e I. furnosoroseus, respectivamente. M. anisopliae apresentou maior eficácia e pode ser considerada como um PFE promissor para o desenvolvimento de micoinseticidas contra o biocontrole efetivo de T. granarium.

A comparative study on the virulence of entomopathogenic fungi against Trogoderma granarium (Everts) (Coleoptera: Dermestidae) in stored grains rice / Um estudo comparativo da virulência de fungos entomopatogênicos contra Trogoderma granarium (Everts) (Coleoptera: Dermestidae) em grãos armazenados de arroz

Entomopathogenic fungi (EPF) now a possible safer microbial control measure that could be considered as a substitute for chemical control of insect pests. Three EPF viz., Metarihizium anisopliae, Isaria furnosoroseus and Beauveria bassiana were evaluated for their virulence against the grubs of Khapra beetle, Trogoderma granarium (Everts) under laboratory conditions. The isolates were applied by two methods viz., diet incorporation and an immersion method with 3rd instar 20 grubs of T. granarium for each. The virulence of EPF was determined using percent mortality. Significantly higher mortality was observed in M. anisopliae applied through immersion (98.33%) and diet incorporation (93.33%) methods followed by B. bassiana (90.83 and 85.83%, respectively). The mortality caused by I. furnosoroseus was statistically lower in immersion and diet incorporation methods i.e. 81.67 and 73.33%, respectively. Based on the immersion method, all EPF were studied for multiple conidial concentration i.e., 1×104, 1×105, 1×106, 1×107 and 1×108 under the same in-vitro conditions. All the isolates were pathogenic to grub of T. granarium at the highest conidial concentration. M. anisopliae was proved the most effective virulent resulting in 98.33% mortality of the pest with LT50 4.61 days at 1 × 108 conidial concentration followed by 90.83 and 81.67 percent mortality with 5.07 and 8.01 days LT50, in the application of B. bassiana and I. furnosoroseus, respectively. M. anisopliae showed higher efficacy and could be considered as promising EPF for the development of myco-insecticides against effective biocontrol of T. granarium.

Os fungos entomopatogênicos (FPE) são agora a possível medida de controle microbiano mais segura, que pode ser considerada um substituto para o controle químico de pragas de insetos. Três EPF viz., Metarihizium anisopliae, Isaria furnosoroseus e Beauveria bassiana foram avaliados quanto à sua virulência contra as larvas do besouro Khapra, Trogoderma granarium (Everts) em condições de laboratório. Os isolados foram aplicados por dois métodos, a saber: incorporação de dieta e um método de imersão com 20 larvas de T. granarium de 3º ínstar para cada um. A virulência do EPF foi determinada usando a mortalidade percentual. Mortalidade significativamente maior foi observada em M. anisopliae aplicado pelos métodos de imersão (98,33%) e incorporação de dieta (93,33%), seguido por B. bassiana (90,83% e 85,83%, respectivamente). A mortalidade causada por I. furnosoroseus foi estatisticamente menor nos métodos de imersão e incorporação de dieta, ou seja, 81,67% e 73,33%, respectivamente. Com base no método de imersão, todos os EPFs foram estudados para múltiplas concentrações de conídios, ou seja, 1 × 104, 1 × 105, 1 × 106, 1 × 107 e 1 × 108 nas mesmas condições in vitro. Todos os isolados foram patogênicos à larva de T. granarium na maior concentração de conídios. M. anisopliae provou ser o virulento mais eficaz, resultando em 98,33% de mortalidade da praga com LT50 4,61 dias na concentração de 1 × 108 conídios seguido por 90,83% e 81,67% de mortalidade com 5,07 e 8,01 dias LT50, na aplicação de B. bassiana e I. furnosoroseus, respectivamente. M. anisopliae apresentou maior eficácia e pode ser considerada como um PFE promissor para o desenvolvimento de micoinseticidas contra o biocontrole efetivo de T. granarium.

A comparative study on the virulence of entomopathogenic fungi against Trogoderma granarium (Everts) (Coleoptera: Dermestidae) in stored grains rice

Abstract Entomopathogenic fungi (EPF) now a possible safer microbial control measure that could be considered as a substitute for chemical control of insect pests. Three EPF viz., Metarihizium anisopliae, Isaria furnosoroseus and Beauveria bassiana were evaluated for their virulence against the grubs of Khapra beetle, Trogoderma granarium (Everts) under laboratory conditions. The isolates were applied by two methods viz., diet incorporation and an immersion method with 3rd instar 20 grubs of T. granarium for each. The virulence of EPF was determined using percent mortality. Significantly higher mortality was observed in M. anisopliae applied through immersion (98.33%) and diet incorporation (93.33%) methods followed by B. bassiana (90.83 and 85.83%, respectively). The mortality caused by I. furnosoroseus was statistically lower in immersion and diet incorporation methods i.e. 81.67 and 73.33%, respectively. Based on the immersion method, all EPF were studied for multiple conidial concentration i.e., 1×104, 1×105, 1×106, 1×107 and 1×108 under the same in-vitro conditions. All the isolates were pathogenic to grub of T. granarium at the highest conidial concentration. M. anisopliae was proved the most effective virulent resulting in 98.33% mortality of the pest with LT50 4.61 days at 1 × 108 conidial concentration followed by 90.83 and 81.67 percent mortality with 5.07 and 8.01 days LT50, in the application of B. bassiana and I. furnosoroseus, respectively. M. anisopliae showed higher efficacy and could be considered as promising EPF for the development of myco-insecticides against effective biocontrol of T. granarium.

Resumo Os fungos entomopatogênicos (FPE) são agora a possível medida de controle microbiano mais segura, que pode ser considerada um substituto para o controle químico de pragas de insetos. Três EPF viz., Metarihizium anisopliae, Isaria furnosoroseus e Beauveria bassiana foram avaliados quanto à sua virulência contra as larvas do besouro Khapra, Trogoderma granarium (Everts) em condições de laboratório. Os isolados foram aplicados por dois métodos, a saber: incorporação de dieta e um método de imersão com 20 larvas de T. granarium de 3º ínstar para cada um. A virulência do EPF foi determinada usando a mortalidade percentual. Mortalidade significativamente maior foi observada em M. anisopliae aplicado pelos métodos de imersão (98,33%) e incorporação de dieta (93,33%), seguido por B. bassiana (90,83% e 85,83%, respectivamente). A mortalidade causada por I. furnosoroseus foi estatisticamente menor nos métodos de imersão e incorporação de dieta, ou seja, 81,67% e 73,33%, respectivamente. Com base no método de imersão, todos os EPFs foram estudados para múltiplas concentrações de conídios, ou seja, 1 × 104, 1 × 105, 1 × 106, 1 × 107 e 1 × 108 nas mesmas condições in vitro. Todos os isolados foram patogênicos à larva de T. granarium na maior concentração de conídios. M. anisopliae provou ser o virulento mais eficaz, resultando em 98,33% de mortalidade da praga com LT50 4,61 dias na concentração de 1 × 108 conídios seguido por 90,83% e 81,67% de mortalidade com 5,07 e 8,01 dias LT50, na aplicação de B. bassiana e I. furnosoroseus, respectivamente. M. anisopliae apresentou maior eficácia e pode ser considerada como um PFE promissor para o desenvolvimento de micoinseticidas contra o biocontrole efetivo de T. granarium.

Efecto Ecotoxicológico de la Mezcla de Aroclor 1254 y Plomo en el Bioindicador Daphnia magna / Ecotoxicological effect of the mixture of Aroclor 1254 and lead in the bioindicator Daphnia magna / Efeito ecotoxicológico da mistura de aroclor 1254 e chumbo no bioindicador Daphnia magna

Resumen Introducción: el bioensayo con la especie bioindicadora Daphnia magna Strauss, 1820 posibilita obtener el efecto ecotoxicológico agudo individual y en mezcla de diversos tóxicos químicos mediante la concentración letal media (CL50). Objetivo: determinar el efecto ecotoxicológico de la mezcla de aroclor 1254 y del plomo (Pb2+) sobre D. magna. Materiales y Métodos: se midió la CL50 siendo aroclor 1254 > Pb2+, y ulteriormente la concentración sin efecto ecológico (PNEC) para el efecto individual y en mezcla. Se contrastaron los valores de PNEC con la categoría 4 - E1 (lagunas y lagos) y E2 (ríos) del Estándar de Calidad Ambiental Peruano (Decreto Supremo N° 004 2017-MINAM -Perú). Resultados: los valores de CL50 disminuyeron de forma gradual a 48 h de exposición para ambos compuestos en forma individual y en mezcla. Se calculó la proporción - sinergia (PS) en relaciones de 1:1, 3:2 y 4:1 (aroclor 1254: Pb2+), obteniéndose mayores valores de PS en 3:2 y 4:1 en contrastación a 1:1. Conclusiones: la concentración de aroclor 1254 estimada de PNEC es similar al valor de la conservación del ambiente acuático categoría 4 - E1 y E2, y la concentración máxima para Pb2+ no es adecuada y debe reducirse para fortalecer la conservación de la biota acuática. Finalmente, el bioindicador D. magna determinó idóneamente la mezcla de aroclor 1254 y Pb2+.

Abstract Introduction: the bioassay with the bioindicator species Daphnia magna Strauss, 1820 makes it possible to obtain the individual acute ecotoxicological effect and in a mixture of various chemical toxics by means of the mean lethal concentration (LC50). Objective: to determine the ecotoxicological effect of the mixture of aroclor 1254 and lead (Pb2+) on D. magna. Materials and Methods: LC50 was measured with arochlor 1254> Pb2+, and subsequently the concentration without ecological effect (PNEC) for the individual effect and as a mixture. PNEC values were contrasted with category 4-E1 (lagoons and lakes) and E2 (rivers) of the Peruvian Environmental Quality Standard (Supreme Decree No. 004 2017-MINAM-Peru). Results: LC50 values gradually decreased after 48 h of exposure for both compounds individually and as a mixture. The proportion-synergy (PS) was calculated in ratios of 1:1, 3:2 and 4:1 (arochlor 1254: Pb2+), obtaining higher PS values in 3:2 and 4:1 in contrast to 1:1. Conclusions: the PNEC estimated arochlor 1254 concentration is similar to the conservation value of the aquatic environment category 4-E1 and E2, and the maximum concentration for Pb2+ is not adequate and should be reduced to strengthen the conservation of aquatic biota. Finally, the bioindicator D. magna ideally determined the mixture of aroclor 1254 and Pb2+.

Resumo Introdução: o bioensaio com as espécies bioindicadoras Daphnia magna Strauss, 1820 permite obter o efeito ecotoxicológico agudo individual e numa mistura de várias toxinas químicas por meio da concentração letal média (CL50). Objetivo: determinar o efeito ecotoxicológico da mistura de arocloro 1254 e chumbo (Pb2+) sobre D. magna. Materiais e Métodos: a CL50 foi medida com arocloro 1254>Pb2+, e posteriormente a concentração sem efeito ecológico (PNEC) para o efeito individual e como mistura. Os valores do PNEC foram contrastados com a categoria 4-E1 (lagoas e lagos) e E2 (rios) do Padrão de Qualidade Ambiental Peruano (Decreto Supremo n° 004 2017-MINAM-Peru). Resultados: Os valores de LC50 diminuíram gradualmente após 48 h de exposição para ambos os compostos individualmente e como uma mistura. A proporção-sinergia (PS) foi calculada em proporções de 1:1, 3:2 e 4:1 (arocloro 1254: Pb2+), obtendo-se valores de PS mais altos em 3:2 e 4:1 em contraste com 1:1. Conclusões: a concentração estimada de arocloro 1254 do PNEC é semelhante ao valor de conservação do ambiente aquático categoria 4-E1 e E2, e a concentração máxima de Pb2+ não é adequada e deve ser reduzida para fortalecer a conservação da biota aquática. Finalmente, o bioindicador D. magna determinou idealmente a mistura de arocloro 1254 e Pb2+.

Acaricidal activity of Furcraea foetida leaf extract against engorged female Rhipicephalus (Boophilus) microplus ticks

The Rhipicephalus (Boophilus) microplus tick is a major concern for the livestock market worldwide, as it causes serious economic damage. Plant-derived acaricides are an attractive alternative to control this ectoparasite and limit the development of resistance. Therefore, the aim of this study was to evaluate the acaricidal activity of Furcraea foetida leaf extract against engorged female R. (B.) microplus ticks. Our in vitro bioassays showed that the crude extract of leaves from F. foetida caused hemorrhagic swelling and skin lesions in the ticks, and three days of treatment caused 100% mortality. Dose-response assay indicated that this toxicity effect was dose-dependent. Similar effects were observed when the crude extract from F. foetida leaves was denatured by boiling at 100°C. These results suggest that the toxicity of the leaf extract might be associated with thermostable biomolecules. Together, our results show for the first time that the crude extract of F. foetida leaves has acaricidal activity against engorged female R. (B.) microplus ticks and it acts in a dose-dependent manner.

Molecular characterization of Toxoplasma gondii isolates from free-range chickens reveals new genotypes in Goiânia, Goiás, Brazil / Caracterização molecular de Toxoplasma gondii isolados de galinhas caipiras revela novos genótipos em Goiânia, Goiás, Brasil

Abstract The aim of this study was to evaluate the genotypic characteristics of Toxoplasma gondii isolated from free-range chickens in the metropolitan area of Goiânia, Goiás, in Brazil's central-west region. The seroprevalence rate was found to be 96%, according to an indirect hemagglutination assay. Brain and heart samples were processed by peptic digestion for a mice bioassay. The tissues were homogenized and the resulting samples were subjected to polymerase chain reaction (PCR), which revealed that 64% of them contained the parasite's DNA. The mice bioassay revealed 15 isolates, 8 of them tachyzoites isolates from the peritoneal lavage and 7 from brain cysts. T. gondii genotypes were determined through PCR-RFLP, using the following markers: SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, alt. SAG2, Apico and CS3. Three genotypes were identified, inclued ToxoDB #65, and the other two are not yet described in the literature. Hence, we conclude that the isolates obtained from the metropolitan area of Goiânia showed relatively low genetic diversity.

Resumo O objetivo deste estudo foi avaliar as características genotípicas de Toxoplasma gondii isolados de galinhas caipiras da Região Metropolitana de Goiânia, Goiás, Região Centro Oeste do Brasil. A soroprevalência foi de 96% dos animais, determinada por hemaglutinação indireta. As amostras de cérebro e coração foram processadas através da digestão péptica para o bioensaio em camundongos. Os tecidos foram homogeneizados, e as amostras resultantes foram analisadas por reação em cadeia da polimerase (PCR), que possibilitou a detecção do DNA do parasito em 64% deles. Por meio do bioensaio em camundongos, foi possível detectar 15 isolados, 8 deles apresentando taquizoítos na lavagem peritoneal e 7 apresentando cistos cerebrais. A determinação dos genótipos de T. gondii foi realizada por PCR-RFLP com os seguintes marcadores: SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, alt. SAG2, Apico e CS3. Foi possível definir 3 genótipos, incluindo o ToxoDB # 65 e dois deles ainda não foram descritos na literatura. Portanto, conclui-se que os isolados obtidos na região metropolitana de Goiânia apresentaram diversidade genética relativamente baixa.

Establishment of high-resolution bioassay profiling platform to screen α-glucosidase inhibitors from Malus hupehensis / 药学学报

italic>α-Glucosidase inhibitors play an important role in the treatment of diabetes. This study established a high-resolution bioassay profiling platform for rapidly screening α-glucosidase inhibitors in natural product extracts. Five α-glucosidase inhibitors were identified from Malus hupehensis, namely, 3-hydroxyphloridzin, quercetin-3-O-β-D-glucopyranoside, phloridzin, avicularin and quercitrin. The establishment and successful application of this platform provides a powerful tool for the efficient discovery of anti-diabetic active ingredients in complex systems.

Toxoplasma gondii genotyping from free-range chickens (Gallus gallus domesticus) in a rural area of Rio Grande do Sul, Brazil / [Genotipagem de Toxoplasma gondii em galinhas domésticas em uma área rural do Rio Grande do Sul, Brasil]

Free-range chickens may ingest oocysts of T. gondii present in the environment and consequently harbor virulent strains of this parasite in different tissues, without any clinical signs. Isolation of T. gondii through bioassays on mice and cats from naturally infected chicken tissues has been described in several countries, demonstrating the importance of free-range chickens in the transmission of this parasite. The aim of this study was the genotypic characterization of T. gondii isolates obtained from naturally infected free-range chickens in a rural area of the state of Rio Grande do Sul, Brazil. Brain and heart tissue from 12 chickens seropositive for T. gondii were processed using peptic digestion technique for parasite isolation. From 12 samples subjected to mouse bioassay, nine isolates were obtained. RFLP-PCR genotypic characterization was performed using 11 genetic markers: SAG1, 5'-3'SAG2, alt.SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1 and Apico. Genetic characterization of the isolates revealed the presence of five atypical genotypes according to ToxoDB (# 11, # 55, # 64, # 140 and # 163). Our results showed a wide genetic diversity of T. gondii in free-range chickens in this region.(AU)

Galinhas criadas ao ar livre podem ingerir oocistos de T. gondii presentes no ambiente e, com isso, albergar cepas virulentas desse parasita em diferentes tecidos, sem sinais clínicos. O isolamento de T. gondii por meio de bioensaios em camundongos e gatos, a partir de tecidos de galinhas naturalmente infectadas, tem sido descrito em vários países. Isso demonstra a importância das galinhas caipiras na epidemiologia desse parasita. O objetivo deste trabalho foi caracterizar genotipicamente isolados de T. gondii obtidos de galinhas caipiras naturalmente infectadas em uma área rural do município de Santa Maria, estado do Rio Grande do Sul, Brasil. Fragmentos de cérebro e de coração, de 12 galinhas soropositivas para T. gondii, foram processados pela técnica de digestão péptica para isolamento do parasita. Das 12 amostras submetidas a bioensaio com camundongos, nove isolados foram obtidos. A caracterização genotípica por RFLP-PCR foi realizada utilizando-se 11 marcadores genéticos: SAG1, 5'-3'SAG2, alt.SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1 e Apico e revelou a presença de cinco genótipos atípicos de acordo com o ToxoDB (# 11, # 55, # 64, # 140 e # 163). Os resultados mostraram uma ampla diversidade genética de T. gondii em galinhas caipiras nessa região.(AU)

Evaluación antimicrobiana de extractos obtenidos a partir de tithonia diversifolia (hemsl.) a. gray con uso potencial en la industria lechera

El uso de antibióticos en el tratamiento de la mastitis en la industria lechera ha acarreado una problemática con potencial impacto en la salud pública, como la resistencia antimicrobiana. El objetivo del presente trabajo fue evaluar la acción antimicrobiana del extracto alcohólico de hojas, tallos y flores de Tithonia diversifolia sobre el crecimiento de Staphylococcus aureus y Escherichia coli en cultivo in vitro. Se evaluó el efecto inhibitorio en el crecimiento bacteriano a través de un ensayo de sensibilidad. En ambas bacterias el control positivo presentó un halo de inhibición superior a 30 mm, con diferencia significativa de los extractos de T. diversifolia. Los cuales presentaron para S. aureus un halo de 9,7 y 10,4 mm y para E. coli de 6,5 y 9,4 mm para tallo-hoja y flores respectivamente. El extracto alcohólico de flores de T. diversifolia se perfila como una herramienta potencial para inhibir el crecimiento bacteriano.

Antibiotics used as a treatment for mastitis in the dairy industry have brought a problem with a potential impact on public health, such as antimicrobial resistance. The objective of this study is to evaluate the antimicrobial action of alcoholic extract from Tithonia diversifolia's leaves, stems, and flowers on the growth of Staphylococcus aureus and Escherichia coli in in vitro cultures. The inhibitory effect on bacterial growth was evaluated through a sensitivity test. In both bacteria, the positive control presented an inhibition halo greater than 30 mm, with a significant difference from the extracts of T. diversifolia, which presented, for S. aureus, a halo of 9.7 and 10.4 mm and for E. coli of 6.5 and 9.4 mm for stem-leaf and flowers respectively. The alcoholic extract of T. diversifolia flowers is emerging as a potential tool to inhibit bacterial growth.

Estimation of sulfentrazone sorption in isolated application and mixed with glyphosate / Estimativa da sorção do sulfentrazone na aplicação isolada e em mistura com glifosato

The use of herbicides belonging to a single mechanism of action for several consecutive years in the same area may lead to the selection of weed biotypes resistant to herbicides. Weeds resistant to glyphosate have been problem worldwide. As an alternative control, farmers have used glyphosate mixed with sulfentrazone. When two herbicides are mixed, the parameters that govern the behaviour of herbicides in the soil can be altered. In this study, the sorption of sulfentrazone was estimated when applied in isolation with glyphosate formulations. For this, increasing doses of sulfentrazone were applied using Red-Yellow Latosol substrate and washed sand. Each dose of sulfentrazone was applied in a mixture with 1080 g a. e. ha-1 Roundup Ready®, Roundup Ultra® and Zapp Qi®. To evaluate the presence of sulfentrazone in the soil solution, Sorghum bicolor was used as an indicator species. Symptoms of intoxication were evaluated at 7, 14 and 21 days after sowing. At 21 days after sowing, the plants were collected and dried in an oven at 70 ± 10 °C to determine the dry matter. Based on the intoxication of the indicator plants, it was verified that, regardless of the formulation, the presence of glyphosate in the mixture increased the sorption of sulfentrazone in the Red-Yellow Latosol. However, based on dry matter accumulation, sulfentrazone sorption increased because of the mixture with Roundup Ultra®, but decreased due to the presence of Roundup Ready® and Zapp Qi®. Therefore, the application of sulfentrazone in mixture with glyphosate alters the sorptive forces of sulfentrazone in a Red-Yellow Latosol.

A utilização de herbicidas pertencentes a um único mecanismo de ação por vários anos consecutivos na mesma área pode levar a seleção de biótipos de plantas daninhas resistentes a herbicidas. Plantas daninhas resistentes ao glyphosate têm sido problema no mundo inteiro. Como alternativa de controle, os agricultores têm utilizado glyphosate em mistura com sulfentrazone. Quando se misturam duas moléculas herbicidas, os parâmetros que governam o comportamento de herbicidas no solo podem ser alterados. Neste estudo foi estimada a sorção do sulfentrazone quando aplicado de forma isolada e em mistura com formulações de glyphosate. Para isso, doses crescentes de sulfentrazone foram aplicadas utilizando-se como substrato Latossolo Vermelho-Amarelo e areia lavada. Cada uma das doses de sulfentrazone foi aplicada em mistura com 1080 g ha-1 de glyphosate comercializado nas formulações Roundup Ready®, Roundup Ultra® e Zapp Qi®. Para avaliar a presença do sulfentrazone na solução do solo utilizou-se o Sorghum bicolor como espécie indicadora. Os sintomas de intoxicação foram avaliados aos 7, 14 e 21 dias após a semeadura. Aos 21 dias após a semeadura, as plantas foram coletadas e secadas em estufa a 70 ±10C para determinação da matéria seca. Com base na intoxicação das plantas indicadoras, verificou-se que, independente da formulação, a presença glyphosate na mistura, aumentou a sorção do sulfentrazone no Latossolo Vermelho-Amarelo. No entanto com base no acúmulo de matéria seca, constatou-se que a sorção do sulfentrazone aumentou em função da mistura com Roundup Ultra®, mas diminuiu devido a presença de Roundup Ready® e Zapp Qi®. Conclui-se que a aplicação de sulfentrazone em mistura com glyphosate altera as forças sortivas do sulfentrazone no Latossolo Vermelho-Amarelo.

Assessing Cosmeceuticals Properties Of Some Macrofungi For Improved Skincare

Objective: This study has investigated cosmeceutical properties namely, antibacterial, sun protection factor and total phenolics contents of some selected macrofungi. The studies were conducted on five reference cultures collected from Indian type culture collection center (IMTEC, Chandigarh, India) and five isolates collected from TERI-Deakin Nano Biotechnology Centre facility at TERI Gram, Gurugram, India. Methods: The cosmeceutical properties of the crude extracts from selected macrofungi were analyzed using standard bioassay techniques. Antibacterial activity was analyzed against Staphylococcus epidermidis, Escherichia coli, Micrococcus luteus, Bacillus megaterium, Pseudomonas aeruginosa, Staphylococcus aureus, and Acinetobacter baumannii using Agar well diffusion method. The sun protection factor was estimated and calculated using the Mansur equation. Free radical scavenging activity using DPPH was performed to assess the antioxidant activity of the extracts. Results: Ethyl acetate extracts of the broth from P. florida and TERI-G1 cultures showed a broad-spectrum antibacterial activity against S. epidermidis, E. coli, M. luteus and B. megaterium. Ethyl acetate extracts of the broth from TERI-G3 showed the highest SPF activity of 34.02 at 200 µg. ml-1 concentration. Ethyl acetate extract of the broth from F. velutipes, P. florida, P. ostreatus, and TERI-G1 showed comparable antioxidant activity of 66.86%, 79.51%, 82.02%, and 69.58% respectively when compared to ascorbic acid (85.83%) and quercetin (83.09%) taken as positive control in the study and their total phenolic contents were found to be 6.93, 43.68, 20.88 and 13.77 Gallic acid equivalent (GAE) per gram. The minimal inhibitory concentration of F. velutipes, P. florida, P. ostreatus, and TERI-G1 was found to be 3552.89 µg. ml-1, 1250 µg. ml-1, 2418.9 µg. ml-1 and 3219 µg. ml-1 respectively. Conclusion: The work is in progress to identify and characterize TERI-G1and TERI G3 cultures. Further studies on the anti-inflammatory, anti-tyrosinase, elastase inhibition properties of the cultures will be assessed to identify potential cosmeceutical active ingredients with promising applications in cosmeceutical products.

Health State, morphological characterization and biometric parameters in the mangrove crab Ucides cordatus Linnaeus, 1763 (Brachyura: Ocypodidae)

The crab Ucides cordatus is an important feature of mangroves in the North and Northeast of Brazil. In 2002 a large mortality was reported on this species in the Jaguaribe River Estuary. In order to investigate the possible causes specimens were collected monthly between May 2002 and December 2003 at five sites: Fortim (E1), Fortim (E2), Cumbe (E3) Aracati (E4) and Guajiru (E5). The mangrove structure and environmental parameters (salinity, pH, temperature and DO) were also evaluated. Water was collected for suspended solids analysis, BOD and nutrient levels. Salinity decreased in the rainy season, with no significant variation of pH and temperature. Dissolved oxygen levels had a mean of 5.7 mg L-1. Suspended solids increased at site E4, with BOD varying at E1 and E2 during the dry season. The same was observed at sites E1, E2 and E3 during the rainy season. Ammonia levels at E1 and E2 and phosphorus in all seasons, increased with rain events. Four lines of investigation were adopted: biometry, histology, hematology, and bioassay. There was no significant difference between animals in biometrics and histology showed no cellular alterations. However, hematology showed a significant difference between the E1 and E3 sites due to depletion in the number of hemocytes in E1, probably in response to environmental impacts. This can lead to poor immunity, leading to opportunistic pathogen infections such as viruses, bacteria and fungi. The bioassay showing no abnormal behavior or mortality. The structure the of mangrove was developed in all areas, except in E5 which served as a control site, with significant environmental stress with high levels of herbivorous growth (> 50%) and a salinity of around 50 ‰. It is believed that the mortality of the crabs was an occasional occurrence, probably due to toxin production by some fungal organism.

Study of Expression of Indigenous Bt cry2AX1 Gene in T3 Progeny of Cotton and its Efficacy Against Helicoverpa armigera (Hubner)

Abstract Development of transgenic Bt crops with stable and high level of Bt protein expression over generations under different environmental conditions is critical for successful deployment at field level. In the present study, progenies of transgenic cotton Coker310 event, CH12 expressing novel cry2AX1 gene were evaluated in T3 generation for stable integration, expression and resistance against cotton bollworm, Helicoverpa armigera. The cry2AX1 gene showed stable inheritance and integration in the T3 progeny plants as revealed by PCR and Southern blot hybridization. The expression of Cry2AX1 protein on 90 days after sowing (DAS) was in the range of 1.055 to 1.5 µg/g of fresh leaf tissue except one plant which showed 0.806 µg/g of fresh leaf tissue and after 30 days (i.e., on 120 DAS) three plants recorded in between 0.69 to 0.82 µg/g and other plants are in range of 0.918 to 1.058 µg/g of fresh leaf tissue. Detached leaf bit bioassay in T3 progeny on 110 DAS recorded mortality of 73.33 to 93.33 per cent against H. armigera and severe growth retardation in surviving larvae. These results indicate that the expression of chimeric cry2AX1 is stable and exhibits insecticidal activity against H. armigera in T3 progeny of CH12 event of transgenic cotton.

Bioassay of anti-gel bioactivity of Chuanxiong Rhizoma and related Chinese patent medicines / 中草药

Objective: Owing to fibrinogen dosage, this study is to develop a method for the bioassay of anti-gel bioactivity of Chuanxiong Rhizoma in vitro, and evaluate the antithrombotic effect of Chuanxiong Rhizoma and its related Chinese patent medicines. Methods: Chuanxiong Rhizoma powder was extracted in ethanol and water, respectively. The mixed extract was used as sample to quantify the level of fibrinogen when the stable fibrin gel was formed in vitro. The fibrinogen dosage was chosen as biomarker for anti-gel bioactivity. Sodium ferulate was chosen as reference. The anti-gel bioactivity of Chuanxiong Rhizoma was calculated according to the direct measurement belong to the bioassay statistical method in the Chinese Pharmacopoeia 2015 (Vol. 4). Moreover, the amounts of anti-gel bioactivity were quantified in nine Chuanxiong Rhizoma samples including Chuanxiong Rhizoma raw materials, decoction pieces, and related Chinese patent medicines to evaluate the applicability for this developed method. Results: Both sodium ferulate and the extract of Chuanxiong Rhizoma showed significant anti-gel bioactivity (P < 0.01). The RSD for the amounts of anti-gel bioactivity was 4.00% (n = 6) in six replicated tests with the confidence limit rate of 7.82% (n = 6). The amounts of anti-gel bioactivity were significant difference among the various types of Chuanxiong Rhizoma samples, i.e. 0.72-1.14 U/g for five Chuanxiong Rhizoma raw materials, 0.68 and 1.32 U/g for Chuanxiong Rhizoma decoction pieces and processed slice with yellow wine, 2.56 and 2.51 U/g for Tongmai Granules and Xuefu Zhuyu Tablets. Conclusion: The developed method can accurately quantify the level of anti-gel bioactivity in Chuanxiong Rhizoma raw materials, decoction pieces and related Chinese patent medicines to assess their antithrombotic effect.

Advances in chromatography-based methods for screening active compounds from natural products / 药学学报

Natural products have been a major source of leading compounds in drug discovery. How to effectively screen active compounds from complex matrix remains an interesting topic. In this review, we comprehensively summarized advanced liquid chromatography based approaches in natural products screening, including pre-column, on-column and post-column screening methods. Their advantages, disadvantages and prospect are also discussed.

Sistema microbiológico de cinco bioensayos para la detección de residuos de antibióticos / Five-assay microbiological system for the screeningf antibiotic residues

Abstract A novel microbiological system in microtiter plates consisting of five bioassays is presented for the detection and classification of antibiotic residues in milk. The bioassays were optimized for the detection of beta-lactams (Bioassay B: Geobacillus stearothermophilus), macrolides (Bioassay M: Bacillus megaterium with fusidic acid), tetracyclines (Bioassay T: B. megaterium with chloramphenicol), quinolones (Bioassay Q: Bacillus licheniformis) and sulfamides (Bioassay QS: B. licheniformis with trimethoprim) at levels near the maximum residue limits (MRL). The response of each bioassay was interpreted visually (positive or negative) after 4-5.5h of incubation. The system detects and classifies beta-lactams (5 pg/l of amoxicillin, 4 pg/l of ampicillin, 36 pg/l of cloxacillin, 22 pg/l of amoxicillin, 3 pg/l of penicillin, 114 pg/l of cephalexin, 89pg/l of cefoperazone and 116 pg/l of ceftiofur), tetracyclines (98 pg/l of chlortetracycline, 92 pg/l of oxytetracycline and 88 pg/l of tetracycline), macrolides (33 pg/l of erythromycin, 44 pg/l of tilmicosin and 50 pg/l of tylosin), sulfonamides (76 pg/l of sulfadiazine, 85 pg/l of sulfadimethoxine, 77 pg/l of sulfamethoxazole and 87pg/l of sulfathiazole) and quinolones (94 pg/l of ciprofloxacin, 98 pg/l of enrofloxacin and 79 pg/l marbofloxacin). In addition, the specificity values were high for B, T, Q (99.4%), M (98.8%) and QS (98.1%) bioassays. The control of antibiotics through this system can contribute to improving the quality and safety of dairy products.

Resumen Se presenta un novedoso sistema microbiológico en placas de microtitulación compuesto por 5 bioensayos para la detección y clasificación de residuos de antibióticos en leche. Los bioensayos fueron optimizados para la detección de betalactámicos (bioensayo B: Geobacillus stearothermophilus), macrólidos (bioensayo M: Bacillus megaterium con ácido fusídico), tetraciclinas (bioensayo T: Bacillus megaterium con cloranfenicol), quinolonas (bioensayo Q: Bacillus licheniformis) y sulfamidas (bioensayo QS: Bacillus licheniformis con trimetoprima), a niveles cercanos a los límites máximos de residuos (LMR). La respuesta de cada bioensayo se interpretó visualmente (positiva o negativa) después de 4 a 5,5 h de incubación. El sistema detecta y clasifica betalactámicos (5 pg/l de amoxicilina, 4 pg/l de ampicilina, 36 pg/l de cloxacilina, 22 pg/l de amoxicilina, 3 pg/l de penicilina, 114 pg/l de cefalexina, 89 pg/l de cefoperazona y 116 pg/l de ceftiofur), tetraciclinas (98 pg/l de clortetraciclina, 92 pg/l de oxitetraciclina y 88 pg/l de tetraciclina), macrólidos (33 pg/l de eritromicina, 44 pg/l de tilmi-cosina y 50 pg/l de tilosina), sulfamidas (76 pg/l de sulfadiacina, 85 pg/l de sulfadimetoxina, 77 pg/l de sulfametoxazol y 87 pg/l de sulfatiazol) y quinolonas (94 pg/l de ciprofloxacina, 98 pg/l de enrofloxacina y 79pg/l de marbofloxacina). Además, los valores de especificidad fueron altos para los bioensayos B, T, Q (99,4%), M (98,8%) y QS (98,1%). El control de residuos de antibióticos mediante este sistema puede contribuir a mejorar la calidad e inocuidad de los productos lácteos.