Phenolic compounds and antifungal activity of ethyl acetate extract and methanolic extract from Capsicum chinense Jacq. ripe fruit

Abstract Food loss due to contamination caused by fungi has much impact on agriculture and leads to significant economic losses. Synthetic and natural fungicides have been used for avoiding losses of several food products due to fungal contamination. As a result, species of the genus Capsicum have been used for preserving food because of their chemical compounds with antifungal activity. Therefore, this study aimed at identifying some phenolic compounds found in both ethyl acetate extract (EAE) and methanolic extract (ME) from habanero pepper (C. chinense) ripe fruit by liquid chromatography tandem mass spectrometry with electrospray ionization (LC-ESI-MS/MS) and at evaluating their antifungal activities against fungi Sclerotinia sclerotiorum, Rhizopus stolonifer and Colletotrichum gloeosporioides. Extracts resulted from a sequential process of maceration. Antifungal activity was evaluated by the disk diffusion method (DDM) at the following doses of both diluted extracts: 25 µL, 50 µL, 100 µL and 200 µL. The chemical analysis showed that there were protocatechuic acid, gentisic acid, vanillic acid, kaempferol-3-O-robinobiosideo and naringenin in both extracts. EAE showed high inhibition of mycelial growth at both doses 100µL and 200µL against the three fungi while methanolic exhibited weak activity even at the highest dose under investigation. However, further in-depth studies are needed to reinforce their uses and practical applications to the agricultural field.

Resumo As perdas de alimentos por contaminação causada por fungos são de grande impacto negativo para a agricultura, gerando altos prejuízos econômicos. Para evitar as perdas de diversos produtos alimentícios pela contaminação fúngica são utilizados fungicidas sintéticos e naturais. As espécies do gênero Capsicum são usadas há muitos anos para auxiliar na conservação de alimentos por possuírem substâncias químicas com ação antifúngica entre outras. Neste contexto, o objetivo deste estudo foi identificar alguns compostos fenólicos por cromatografia líquida de alta eficiência acoplada à espectrometria de massas sequencial (LC-ESI-MS/MS) presentes nos extratos acetato de etila (EAE) e metanólico (ME) dos frutos maduros da pimenta biquinho (C. chinense) e avaliar atividade antifúngica de EAE e ME contra os fungos Sclerotinia sclerotiorum, Rhizopus stolonifer e Colletotrichum gloeosporioides. Os extratos foram obtidos de forma sequencial, utilizando o procedimento de maceração. A atividade antifúngica foi avaliada seguindo a metodologia de difusão em disco, nas doses de 25 µL, 50 µL, 100 µL e 200 µL de cada extrato diluído. A análise química evidenciou a presença de ácido protocatequico, ácido gentisico, ácido vanílico, kaempferol-3-O-robinobiosídeo e naringenina em ambos os extratos. EAE revelou maior poder de inibição do crescimento micelial nas doses de 100µL e 200µL contra os três fungos testados, enquanto ME exibiu fraca atividade inclusive na maior dose investigada. Entretanto, estudos mais aprofundados ainda são necessários para consolidar seu uso e aplicação prática na área agronômica.

Phenolic compounds and antifungal activity of ethyl acetate extract and methanolic extract from Capsicum chinense Jacq. ripe fruit / Compostos fenólicos e atividade antifúngica dos extratos acetato de etila e metanólico dos frutos maduros de Capsicum chinense Jacq

Food loss due to contamination caused by fungi has much impact on agriculture and leads to significant economic losses. Synthetic and natural fungicides have been used for avoiding losses of several food products due to fungal contamination. As a result, species of the genus Capsicum have been used for preserving food because of their chemical compounds with antifungal activity. Therefore, this study aimed at identifying some phenolic compounds found in both ethyl acetate extract (EAE) and methanolic extract (ME) from habanero pepper (C. chinense) ripe fruit by liquid chromatography tandem mass spectrometry with electrospray ionization (LC-ESI-MS/MS) and at evaluating their antifungal activities against fungi Sclerotinia sclerotiorum, Rhizopus stolonifer and Colletotrichum gloeosporioides. Extracts resulted from a sequential process of maceration. Antifungal activity was evaluated by the disk diffusion method (DDM) at the following doses of both diluted extracts: 25 µL, 50 µL, 100 µL and 200 µL. The chemical analysis showed that there were protocatechuic acid, gentisic acid, vanillic acid, kaempferol-3-O-robinobiosideo and naringenin in both extracts. EAE showed high inhibition of mycelial growth at both doses 100µL and 200µL against the three fungi while methanolic exhibited weak activity even at the highest dose under investigation. However, further in-depth studies are needed to reinforce their uses and practical applications to the agricultural field.

As perdas de alimentos por contaminação causada por fungos são de grande impacto negativo para a agricultura, gerando altos prejuízos econômicos. Para evitar as perdas de diversos produtos alimentícios pela contaminação fúngica são utilizados fungicidas sintéticos e naturais. As espécies do gênero Capsicum são usadas há muitos anos para auxiliar na conservação de alimentos por possuírem substâncias químicas com ação antifúngica entre outras. Neste contexto, o objetivo deste estudo foi identificar alguns compostos fenólicos por cromatografia líquida de alta eficiência acoplada à espectrometria de massas sequencial (LC-ESI-MS/MS) presentes nos extratos acetato de etila (EAE) e metanólico (ME) dos frutos maduros da pimenta biquinho (C. chinense) e avaliar atividade antifúngica de EAE e ME contra os fungos Sclerotinia sclerotiorum, Rhizopus stolonifer e Colletotrichum gloeosporioides. Os extratos foram obtidos de forma sequencial, utilizando o procedimento de maceração. A atividade antifúngica foi avaliada seguindo a metodologia de difusão em disco, nas doses de 25 µL, 50 µL, 100 µL e 200 µL de cada extrato diluído. A análise química evidenciou a presença de ácido protocatequico, ácido gentisico, ácido vanílico, kaempferol-3-O-robinobiosídeo e naringenina em ambos os extratos. EAE revelou maior poder de inibição do crescimento micelial nas doses de 100µL e 200µL contra os três fungos testados, enquanto ME exibiu fraca atividade inclusive na maior dose investigada. Entretanto, estudos mais aprofundados ainda são necessários para consolidar seu uso e aplicação prática na área agronômica.

Phenolic compounds and antifungal activity of ethyl acetate extract and methanolic extract from Capsicum chinense Jacq. ripe fruit.

Food loss due to contamination caused by fungi has much impact on agriculture and leads to significant economic losses. Synthetic and natural fungicides have been used for avoiding losses of several food products due to fungal contamination. As a result, species of the genus Capsicum have been used for preserving food because of their chemical compounds with antifungal activity. Therefore, this study aimed at identifying some phenolic compounds found in both ethyl acetate extract (EAE) and methanolic extract (ME) from habanero pepper (C. chinense) ripe fruit by liquid chromatography tandem mass spectrometry with electrospray ionization (LC-ESI-MS/MS) and at evaluating their antifungal activities against fungi Sclerotinia sclerotiorum, Rhizopus stolonifer and Colletotrichum gloeosporioides. Extracts resulted from a sequential process of maceration. Antifungal activity was evaluated by the disk diffusion method (DDM) at the following doses of both diluted extracts: 25 µL, 50 µL, 100 µL and 200 µL. The chemical analysis showed that there were protocatechuic acid, gentisic acid, vanillic acid, kaempferol-3-O-robinobiosideo and naringenin in both extracts. EAE showed high inhibition of mycelial growth at both doses 100µL and 200µL against the three fungi while methanolic exhibited weak activity even at the highest dose under investigation. However, further in-depth studies are needed to reinforce their uses and practical applications to the agricultural field.

Chemical composition and in vitro inhibitory effects of essential oils from fruit peel of three Citrus species and limonene on mycelial growth of Sclerotinia sclerotiorum / Composição química e efeito inibitório in vitro dos óleos essenciais das cascas dos frutos de três espécies de Citrus e do limoneno sobre o crescimento micelial de Sclerotinia sclerotiorum

Abstract Essential oils (EO) from aromatic and medicinal plants generally perform a diverse range of biological activities because they have several active constituents that work in different mechanisms of action. EO from Citrus peel have an impressive range of food and medicinal uses, besides other applications. EO from Citrus reticulata, C. sinensis and C. deliciosa were extracted from fruit peel and analyzed by GC-MS. The major constituent of EO under evaluation was limonene, whose concentrations were 98.54%, 91.65% and 91.27% for C. sinensis, C. reticulata and C. deliciosa, respectively. The highest potential of inhibition of mycelial growth was observed when the oil dose was 300 μL. Citrus oils inhibited fungus growth in 82.91% (C. deliciosa), 65.82% (C. sinensis) and 63.46% (C. reticulata). Anti-Sclerotinia sclerotiorum activity of 90% pure limonene and at different doses (20, 50, 100, 200 and 300 μL) was also investigated. This monoterpene showed to be highly active by inhibiting 100% fungus growth even at 200 and 300 μL doses. This is the first report of the in vitro inhibitory effect of natural products from these three Citrus species and its results show that there is good prospect of using them experimentally to control S. sclerotiorum, in both greenhouse and field conditions.

Resumo Óleos essenciais de plantas aromáticas e medicinais geralmente exibem uma gama diversificada de atividades biológicas, pois possuem vários constituintes ativos que atuam por meio de vários mecanismos de ação. Os óleos essenciais das cascas de Citrus têm uma variedade impressionante de usos em alimentos, medicamentos entre várias outras aplicações. Os óleos essenciais (OE) de Citrus reticulata, C. sinenses e C. deliciosa foram extraídos das cascas dos frutos e analisados por CG-EM. O limoneno foi o constituinte majoritário encontrado nos óleos essenciais avaliados, nas concentrações de 98,54%, 91,65% e 91,27% para C. sinensis, C. reticulata e C. deliciosa, respectivamente. Os maiores potenciais de inibição do crescimento micelial foi observado na dose de 300 µL dos óleos. Os óleos de Citrus inibiram em 82,91% (C. deliciosa), 65,82% (C. sinensis) e 63,46% (C. reticulata) o crescimento do fungo. A atividade anti-Sclerotinia sclerotiorum do limoneno 90% puro e em diferentes doses (20, 50, 100, 200 e 300 µL) foi também investigada e este monoterpeno demonstrou-se altamente ativo inibindo 100% o crescimento do fungo inclusive nas doses de 200 e 300 µL. Este é o primeiro relato sobre o efeito inibitório in vitro dos óleos essenciais destas três espécies de Citrus e os resultados deste estudo mostram que existe uma boa perspectiva de uso destes produtos naturais experimentalmente para controlar o S. sclerotiorum tanto em condições de estufa como em condições de campo.

Chemical composition and in vitro inhibitory effects of essential oils from fruit peel of three Citrus species and limonene on mycelial growth of Sclerotinia sclerotiorum.

Essential oils (EO) from aromatic and medicinal plants generally perform a diverse range of biological activities because they have several active constituents that work in different mechanisms of action. EO from Citrus peel have an impressive range of food and medicinal uses, besides other applications. EO from Citrus reticulata, C. sinensis and C. deliciosa were extracted from fruit peel and analyzed by GC-MS. The major constituent of EO under evaluation was limonene, whose concentrations were 98.54%, 91.65% and 91.27% for C. sinensis, C. reticulata and C. deliciosa, respectively. The highest potential of inhibition of mycelial growth was observed when the oil dose was 300 µL. Citrus oils inhibited fungus growth in 82.91% (C. deliciosa), 65.82% (C. sinensis) and 63.46% (C. reticulata). Anti-Sclerotinia sclerotiorum activity of 90% pure limonene and at different doses (20, 50, 100, 200 and 300 µL) was also investigated. This monoterpene showed to be highly active by inhibiting 100% fungus growth even at 200 and 300 µL doses. This is the first report of the in vitro inhibitory effect of natural products from these three Citrus species and its results show that there is good prospect of using them experimentally to control S. sclerotiorum, in both greenhouse and field conditions.

Chemical composition and in vitro inhibitory effects of essential oils from fruit peel of three Citrus species and limonene on mycelial growth of Sclerotinia sclerotiorum / Composição química e efeito inibitório in vitro dos óleos essenciais das cascas dos frutos de três espécies de Citrus e do limoneno sobre o crescimento micelial de Sclerotinia sclerotiorum

Essential oils (EO) from aromatic and medicinal plants generally perform a diverse range of biological activities because they have several active constituents that work in different mechanisms of action. EO from Citrus peel have an impressive range of food and medicinal uses, besides other applications. EO from Citrus reticulata, C. sinensis and C. deliciosa were extracted from fruit peel and analyzed by GC-MS. The major constituent of EO under evaluation was limonene, whose concentrations were 98.54%, 91.65% and 91.27% for C. sinensis, C. reticulata and C. deliciosa, respectively. The highest potential of inhibition of mycelial growth was observed when the oil dose was 300 L. Citrus oils inhibited fungus growth in 82.91% (C. deliciosa), 65.82% (C. sinensis) and 63.46% (C. reticulata). Anti-Sclerotinia sclerotiorum activity of 90% pure limonene and at different doses (20, 50, 100, 200 and 300 L) was also investigated. This monoterpene showed to be highly active by inhibiting 100% fungus growth even at 200 and 300 L doses. This is the first report of the in vitro inhibitory effect of natural products from these three Citrus species and its results show that there is good prospect of using them experimentally to control S. sclerotiorum, in both greenhouse and field conditions.(AU)

Óleos essenciais de plantas aromáticas e medicinais geralmente exibem uma gama diversificada de atividades biológicas, pois possuem vários constituintes ativos que atuam por meio de vários mecanismos de ação. Os óleos essenciais das cascas de Citrus têm uma variedade impressionante de usos em alimentos, medicamentos entre várias outras aplicações. Os óleos essenciais (OE) de Citrus reticulata, C. sinenses e C. deliciosa foram extraídos das cascas dos frutos e analisados por CG-EM. O limoneno foi o constituinte majoritário encontrado nos óleos essenciais avaliados, nas concentrações de 98,54%, 91,65% e 91,27% para C. sinensis, C. reticulata e C. deliciosa, respectivamente. Os maiores potenciais de inibição do crescimento micelial foi observado na dose de 300 µL dos óleos. Os óleos de Citrus inibiram em 82,91% (C. deliciosa), 65,82% (C. sinensis) e 63,46% (C. reticulata) o crescimento do fungo. A atividade anti-Sclerotinia sclerotiorum do limoneno 90% puro e em diferentes doses (20, 50, 100, 200 e 300 µL) foi também investigada e este monoterpeno demonstrou-se altamente ativo inibindo 100% o crescimento do fungo inclusive nas doses de 200 e 300 µL. Este é o primeiro relato sobre o efeito inibitório in vitro dos óleos essenciais destas três espécies de Citrus e os resultados deste estudo mostram que existe uma boa perspectiva de uso destes produtos naturais experimentalmente para controlar o S. sclerotiorum tanto em condições de estufa como em condições de campo.(AU)

Blocking and remanence properties of weakly and highly interactive cobalt ferrite based nanoparticles.

We compare both magnetic blocking properties and remanence curves for dilute ferrofluid and powder samples of ferrite magnetic nanoparticles. Low field DC magnetization, AC susceptibility, isothermal remanent magnetization and DC demagnetization techniques are employed to investigate the role of interparticle magnetic interactions on the superparamagnetic relaxation, the magnetic anisotropy and on the super-spin-glass state in closely packed particles. The samples used herein are 3 nm sized spinel-type nanocrystals made of a cobalt ferrite core covered by a layer of maghemite on its outermost surface and can be obtained as aqueous colloidal dispersions thanks to this core-shell strategy. They show large anisotropy attributed to an enhanced surface contribution and the blocking temperature is shifted towards higher values as interparticle distance decreases. For all investigated diluted liquids and powder samples the frequency dependency of the peak temperature is well accounted by a Vogel-Fulcher law, with the insertion of a phenomenological temperature associated to the magnitude of interparticle dipolar interactions. The fractional change of the peak temperature per decade of frequency enlights the presence of interactions between particles in dilute liquids and of a spin-glass-like state in powder samples. The remanence curves always show global demagnetizing behavior, attributed to the combination of both spin surface disorder and interparticle dipolar interactions, the former being predominant in isolated nanoparticles and the latter in powder samples. However, in the most compacted powder, exchange interaction between surface ions of different particles becomes more pronounced and promotes an additive magnetizing effect.

Chemical composition and in vitro inhibitory effects of essential oils from fruit peel of three Citrus species and limonene on mycelial growth of Sclerotinia sclerotiorum

Abstract Essential oils (EO) from aromatic and medicinal plants generally perform a diverse range of biological activities because they have several active constituents that work in different mechanisms of action. EO from Citrus peel have an impressive range of food and medicinal uses, besides other applications. EO from Citrus reticulata, C. sinensis and C. deliciosa were extracted from fruit peel and analyzed by GC-MS. The major constituent of EO under evaluation was limonene, whose concentrations were 98.54%, 91.65% and 91.27% for C. sinensis, C. reticulata and C. deliciosa, respectively. The highest potential of inhibition of mycelial growth was observed when the oil dose was 300 L. Citrus oils inhibited fungus growth in 82.91% (C. deliciosa), 65.82% (C. sinensis) and 63.46% (C. reticulata). Anti-Sclerotinia sclerotiorum activity of 90% pure limonene and at different doses (20, 50, 100, 200 and 300 L) was also investigated. This monoterpene showed to be highly active by inhibiting 100% fungus growth even at 200 and 300 L doses. This is the first report of the in vitro inhibitory effect of natural products from these three Citrus species and its results show that there is good prospect of using them experimentally to control S. sclerotiorum, in both greenhouse and field conditions.

Resumo Óleos essenciais de plantas aromáticas e medicinais geralmente exibem uma gama diversificada de atividades biológicas, pois possuem vários constituintes ativos que atuam por meio de vários mecanismos de ação. Os óleos essenciais das cascas de Citrus têm uma variedade impressionante de usos em alimentos, medicamentos entre várias outras aplicações. Os óleos essenciais (OE) de Citrus reticulata, C. sinenses e C. deliciosa foram extraídos das cascas dos frutos e analisados por CG-EM. O limoneno foi o constituinte majoritário encontrado nos óleos essenciais avaliados, nas concentrações de 98,54%, 91,65% e 91,27% para C. sinensis, C. reticulata e C. deliciosa, respectivamente. Os maiores potenciais de inibição do crescimento micelial foi observado na dose de 300 µL dos óleos. Os óleos de Citrus inibiram em 82,91% (C. deliciosa), 65,82% (C. sinensis) e 63,46% (C. reticulata) o crescimento do fungo. A atividade anti-Sclerotinia sclerotiorum do limoneno 90% puro e em diferentes doses (20, 50, 100, 200 e 300 µL) foi também investigada e este monoterpeno demonstrou-se altamente ativo inibindo 100% o crescimento do fungo inclusive nas doses de 200 e 300 µL. Este é o primeiro relato sobre o efeito inibitório in vitro dos óleos essenciais destas três espécies de Citrus e os resultados deste estudo mostram que existe uma boa perspectiva de uso destes produtos naturais experimentalmente para controlar o S. sclerotiorum tanto em condições de estufa como em condições de campo.

Promising synergistic activity of fluconazole with bioactive Guttiferone-A and derivatives against non-albicans Candida species.

Natural products with diverse bioactivities and structures are an important source of novel chemicals with pharmaceutical potentials. Combinations of bioactive compounds with classical antimicrobial agents against drug-resistant or low-susceptible Candida spp. have been studied. Guttiferone-A and its derivatives were combined with fluconazole through the checkerboard method and tested against Candida spp. The results obtained, especially the Fractional Inhibitory Concentration Index (FICI) determined to the combinations, suggests promising results on the treatment of Candida infections, principally for species that present resistance or low antifungal susceptibility. The best result was seen for C. krusei, in which a synergic action of the association between fluconazole and Guttiferone-A resulted in a reduction of more than 100-fold in the alone inhibitory concentration (MIC) of fluconazole. Synergism was also noted in the association of fluconazole with the synthetic derivatives LFQM-79, LFQM-80 and LFQM-81 against C. glabrata, with reduction of up to four times in the alone IC of fluconazole. These results suggest the possibility of combined administration with reduction of doses and side effects of drugs conventionally used against Candida spp. and the promising therapeutic action of Guttiferone-A.

Chemical composition and in vitro inhibitory effects of essential oils from fruit peel of three Citrus species and limonene on mycelial growth of Sclerotinia sclerotiorum

Abstract Essential oils (EO) from aromatic and medicinal plants generally perform a diverse range of biological activities because they have several active constituents that work in different mechanisms of action. EO from Citrus peel have an impressive range of food and medicinal uses, besides other applications. EO from Citrus reticulata, C. sinensis and C. deliciosa were extracted from fruit peel and analyzed by GC-MS. The major constituent of EO under evaluation was limonene, whose concentrations were 98.54%, 91.65% and 91.27% for C. sinensis, C. reticulata and C. deliciosa, respectively. The highest potential of inhibition of mycelial growth was observed when the oil dose was 300 L. Citrus oils inhibited fungus growth in 82.91% (C. deliciosa), 65.82% (C. sinensis) and 63.46% (C. reticulata). Anti-Sclerotinia sclerotiorum activity of 90% pure limonene and at different doses (20, 50, 100, 200 and 300 L) was also investigated. This monoterpene showed to be highly active by inhibiting 100% fungus growth even at 200 and 300 L doses. This is the first report of the in vitro inhibitory effect of natural products from these three Citrus species and its results show that there is good prospect of using them experimentally to control S. sclerotiorum, in both greenhouse and field conditions.

Resumo Óleos essenciais de plantas aromáticas e medicinais geralmente exibem uma gama diversificada de atividades biológicas, pois possuem vários constituintes ativos que atuam por meio de vários mecanismos de ação. Os óleos essenciais das cascas de Citrus têm uma variedade impressionante de usos em alimentos, medicamentos entre várias outras aplicações. Os óleos essenciais (OE) de Citrus reticulata, C. sinenses e C. deliciosa foram extraídos das cascas dos frutos e analisados por CG-EM. O limoneno foi o constituinte majoritário encontrado nos óleos essenciais avaliados, nas concentrações de 98,54%, 91,65% e 91,27% para C. sinensis, C. reticulata e C. deliciosa, respectivamente. Os maiores potenciais de inibição do crescimento micelial foi observado na dose de 300 µL dos óleos. Os óleos de Citrus inibiram em 82,91% (C. deliciosa), 65,82% (C. sinensis) e 63,46% (C. reticulata) o crescimento do fungo. A atividade anti-Sclerotinia sclerotiorum do limoneno 90% puro e em diferentes doses (20, 50, 100, 200 e 300 µL) foi também investigada e este monoterpeno demonstrou-se altamente ativo inibindo 100% o crescimento do fungo inclusive nas doses de 200 e 300 µL. Este é o primeiro relato sobre o efeito inibitório in vitro dos óleos essenciais destas três espécies de Citrus e os resultados deste estudo mostram que existe uma boa perspectiva de uso destes produtos naturais experimentalmente para controlar o S. sclerotiorum tanto em condições de estufa como em condições de campo.

Role of MTHFR C677T and MTR A2756G polymorphisms in thyroid and breast cancer development.

Folate metabolism is essential for DNA synthesis and repair. Alterations in genes that participate in folate metabolism can be associated with several types of malignant neoplasms, including thyroid and breast cancer. In the present case-control study, we examined the association between methylenetetrahydrofolate reductase (MTHFR C677T, rs1801133) and methionine synthase (MTR A2756G, rs1805087) polymorphisms and risk for thyroid and breast cancer. Polymerase chain reaction-restriction fragment length technique was used to determine the specific genotypes in the genes of interest. Statistical analysis was performed by multiple logistic regression test. We found an association between MTHFR C677T polymorphism and risks to both thyroid (OR = 2.50; 95%CI = 1.15-5.46; P = 0.02) and breast cancer (OR = 2.53; 95%CI = 1.08-5.93; P = 0.03). Tobacco consumption and high body mass index were also associated with thyroid cancer. In addition, increased age (≥50 years) and alcohol consumption were found to be associated with breast cancer. Our results indicated that MTHFR C677T is significantly associated with thyroid and breast cancer risks. Thus, these factors may be used as potential prognostic markers for thyroid and breast cancers.

Sex identification of domestic geese (Anser anser): comparison of cytogenetic analysis technique and post anesthetic inspection of the cloaca / Identificação do sexo de gansos domésticos (Anser anser): comparação da técnica de sexagem cromossômica e a inspeção pós-anestésica da cloaca

The domestic geese used in this experiment were free-living, from a lake located in an urban area and its population had increased dramatically. The need for a project proposing the sterilization of some males to restrain population growth, led to the need for sexing these animals. As domestic geese present minimal sexual dimorphism, two techniques were tested: cytogenetic analysis and post anesthetic inspection of the cloaca. There were used eight adults geese randomly chosen from a group of 80 animals. They were caught with nets or by hand, were individually transported and housed in appropriate place waiting for the cytogenetic analysis (three to four days). For the chromosomal sexing, was collected 2 mL of blood and the peripheral blood lymphocyte culture technique was used. The sample was treated with hypotonic KCl, fixed in three changes of methanol/acetic acid solution. The slides were prepared with the standard technique and examined under an optical microscope to observe the metaphase. To the technique of cloaca eversion, was performed injectable anesthesia with tiletamine-zolazepam 5%. Ten minutes after anesthesia, the cloaca has been everted to visualize the internal structures and sex determination. The techniques were performed in blinded trial to avoidinaccurate results. From the eight animals submitted to chromosomal sexing technique, five were identified as male and three as females, reaching 100% agreement when compared with the cloaca eversion technique. Sexing chromosomal technique appeared to cause the least discomfort in the animals evaluated, proving to be a reliable technique for sexualdetermination and favorable to animal welfare.

Os gansos domésticos utilizados neste experimento eram de vida livre, pertencentes a um lago localizado em uma área urbana e sua população havia aumentado dramaticamente. A necessidade de um projeto com foco na esterilização de alguns machos, para reduzir o crescimento populacional, levou à necessidade de sexagem desses animais. como os gansos domésticos apresentam mínimo dimorfismo sexual, duas técnicas foram testadas: análise citogenética e inspeção da cloaca pós-anestesia. Foram utilizados oito gansos adultos, escolhidos aleatoriamente de um grupo de 80 animais. Eles foram capturados com redes ou manualmente, transportados individualmente e alojados em local apropriado a espera da análise citogenética (três a quatro dias). Para a sexagem cromossomal, foram coletados 2 ml de sangue e a técnica de cultura de linfócitos periféricos foi empregada. As amostras resultantes foram tratadas com kcl hipotônico, fixadas em três trocas de solução de metanol e ácido acético. As lâminas foram preparadas com a técnica padrão e examinadas ao microscópio óptico para observação das metáfases. para a técnica de eversão da cloaca, foi realizada a anestesia injetável com tiletamina-zolazepam 5%. Dez minutos após a anestesia, a cloaca foi evertida para a visualização das estruturas internas e a determinação sexual. AAs técnicas foram conduzidas em duplo cego, para melhor acurácia dos resultados. Dos oito animais submetidos à técnica de sexagem cromossomal, cinco foram identificados como machos e três como fêmeas, alcançando 100% de concordância quando comparada com a técnica de eversão da cloaca. A técnica de sexagem cromossomal aparentou causar o menor desconforto nos animais avaliados, mostrando-se uma técnicaconfiável para determinação sexual e favorável ao bem-estar animal.

IDENTIFICAÇÃO DO SEXO DE GANSOS DOMÉSTICOS (ANSER ANSER): COMPARAÇÃO DA TÉCNICA DE SEXAGEM CROMOSSÔMICA E A INSPEÇÃO PÓS-ANESTÉSICA DA CLOACA

Os gansos domésticos utilizados neste experimento eram de vida livre, pertencentes a um lago localizado em uma área urbana e sua população havia aumentado dramaticamente. a necessidade de um projeto com foco na esterilização de alguns machos, para reduzir o crescimento populacional, levou à necessidade de sexagem desses animais. como os gansos domésticos apresentam mínimo dimorfismo sexual, duas técnicas foram testadas: análise citogenética e inspeção da cloaca pós-anestesia. foram utilizados oito gansos adultos, escolhidos aleatoriamente de um grupo de 80 animais. eles foram capturados com redes ou manualmente, transportados individualmente e alojados em local apropriado a espera da análise citogenética (três a quatro dias). para a sexagem cromossomal, foram coletados 2 ml de sangue e a técnica de cultura de linfócitos periféricos foi empregada. as amostras resultantes foram tratadas com kcl hipotônico, fixadas em três trocas de solução de metanol e ácido acético. as lâminas foram preparadas com a técnica padrão e examinadas ao microscópio óptico para observação das metáfases. para a técnica de eversão da cloaca, foi realizada a anestesia injetável com tiletamina-zolazepam 5%. dez minutos após a anestesia, a cloaca foi evertida para a visualização das estruturas internas e a determinação sexual. As técnicas foram conduzidas em duplo cego, para melhor acurácia dos resultados. D

Sex identification of domestic geese (Anser anser): comparison of cytogenetic analysis technique and post anesthetic inspection of the cloaca / Identificação do sexo de gansos domésticos (Anser anser): comparação da técnica de sexagem cromossômica e a inspeção pós-anestésica da cloaca

The domestic geese used in this experiment were free-living, from a lake located in an urban area and its population had increased dramatically. The need for a project proposing the sterilization of some males to restrain population growth, led to the need for sexing these animals. As domestic geese present minimal sexual dimorphism, two techniques were tested: cytogenetic analysis and post anesthetic inspection of the cloaca. There were used eight adults geese randomly chosen from a group of 80 animals. They were caught with nets or by hand, were individually transported and housed in appropriate place waiting for the cytogenetic analysis (three to four days). For the chromosomal sexing, was collected 2 mL of blood and the peripheral blood lymphocyte culture technique was used. The sample was treated with hypotonic KCl, fixed in three changes of methanol/acetic acid solution. The slides were prepared with the standard technique and examined under an optical microscope to observe the metaphase. To the technique of cloaca eversion, was performed injectable anesthesia with tiletamine-zolazepam 5%. Ten minutes after anesthesia, the cloaca has been everted to visualize the internal structures and sex determination. The techniques were performed in blinded trial to avoidinaccurate results. From the eight animals submitted to chromosomal sexing technique, five were identified as male and three as females, reaching 100% agreement when compared with the cloaca eversion technique. Sexing chromosomal technique appeared to cause the least discomfort in the animals evaluated, proving to be a reliable technique for sexualdetermination and favorable to animal welfare. (AU)

Os gansos domésticos utilizados neste experimento eram de vida livre, pertencentes a um lago localizado em uma área urbana e sua população havia aumentado dramaticamente. A necessidade de um projeto com foco na esterilização de alguns machos, para reduzir o crescimento populacional, levou à necessidade de sexagem desses animais. como os gansos domésticos apresentam mínimo dimorfismo sexual, duas técnicas foram testadas: análise citogenética e inspeção da cloaca pós-anestesia. Foram utilizados oito gansos adultos, escolhidos aleatoriamente de um grupo de 80 animais. Eles foram capturados com redes ou manualmente, transportados individualmente e alojados em local apropriado a espera da análise citogenética (três a quatro dias). Para a sexagem cromossomal, foram coletados 2 ml de sangue e a técnica de cultura de linfócitos periféricos foi empregada. As amostras resultantes foram tratadas com kcl hipotônico, fixadas em três trocas de solução de metanol e ácido acético. As lâminas foram preparadas com a técnica padrão e examinadas ao microscópio óptico para observação das metáfases. para a técnica de eversão da cloaca, foi realizada a anestesia injetável com tiletamina-zolazepam 5%. Dez minutos após a anestesia, a cloaca foi evertida para a visualização das estruturas internas e a determinação sexual. AAs técnicas foram conduzidas em duplo cego, para melhor acurácia dos resultados. Dos oito animais submetidos à técnica de sexagem cromossomal, cinco foram identificados como machos e três como fêmeas, alcançando 100% de concordância quando comparada com a técnica de eversão da cloaca. A técnica de sexagem cromossomal aparentou causar o menor desconforto nos animais avaliados, mostrando-se uma técnicaconfiável para determinação sexual e favorável ao bem-estar animal. (AU)

Fenóis totais, atividade antioxidante e inibição da enzima tirosinase de extratos de Myracrodruon urundeuva Fr. All. (Anacardiaceae) / Total phenolics, antioxidant activity and tyrosinase enzyme inhibition by Myracrodruon urundeuva Fr. All. (Anacardiaceae) extracts

RESUMOOs radicais livres e outros oxidantes demonstram ser parcialmente responsáveis pelo envelhecimento e pelas doenças degenerativas associadas à produção de espécies reativas de oxigênio. Além disto, os compostos fenólicos são reconhecidamente detentores de elevada atividade antioxidante, que geralmente estão envolvidos em tratamentos de problemas de pigmentação da pele, que resultam em hiperpigmentações. Sendo assim este trabalho foi realizado para avaliar o conteúdo de fenóis totais, atividade antioxidante e a capacidade de inibição da tirosinase dos extratos das folhas e cascas do caule da espécie Myracrodruonurundeuva Fr. All. Para determinação do conteúdo de fenóis totais dos extratos orgânicos de M. urundeuva utilizou-se o reativo Folin-Ciocalteau, na avaliação da atividade antioxidante empregando o radical livre DPPH, enquanto que os testes de inibição da enzima tirosinase foram realizados utilizando L-tirosina. A concentração de fenóis totais foi de 77 mg EAG g-1 e 194 mg EAG g-1 nos extratos hexânico e metanólico das folhas e de 45 mg EAG g-1 e 193 mg EAG g-1 nos extratos hexânico e metanólico das cascas do caule. O potencial antioxidante dos extratos indicaram que o extrato metanólico das cascas do caule (10,9 ± 0,5 µg mL-1), em comparação ao hexânico (12,9 ± 0,2 µg mL-1) e ao BHT (220 ± 7,0 µg mL-1), possui atividade antioxidante levemente mais acentuada. No ensaio de inibição da tirosinase, o extrato metanólico das cascas do caule demonstrou a inibição da enzima em 42% após uma hora.

ABSTRACTFree radicals and other oxidants had demonstrated to be partially responsible for aging and for degenerative diseases associated with the production of reactive oxygen species. In addition, the phenolic compounds are recognized as holders of high antioxidant activity, which usually are involved in the treatment of pigmentation problems, resulting in hyper pigmentation. So this study was carried out in order to evaluate the content of total phenols, antioxidant activity and capacity inhibition of tyrosinase in extracts from leaves and stem barks of the species Myracrodruon urundeuva Fr. All. In order to determine the content of total phenols from organic extracts of M. urundeuva, a Folin-Ciocalteau reagent was employed in the evaluation of antioxidant activity,using the DPPH free radical, while the enzyme tyrosinase inhibition tests were performed using L-tyrosine. The amount of total phenols was 77 mg EAG g-1 and 194 in the hexane and methanolic extracts from the leaves and 45 mg EAG g-1 and 193 mg EAG g-1in the hexane and methanolic extracts of the stem barks. The antioxidant potential of extracts indicated that the methanolic extract of stem bark (10.9 ± 0.5 µg mL-1), when compared to the hexane (12.9 ± 0.2 µg mL-1) and BHT (220 ± 7.0 µg mL-1), presented a slightly higher antioxidant activity. For the tyrosinase inhibitiontrial , the methanolic extract ofstem barks indicated anenzyme inhibition of 42% after an hour.

Karyotypic diversity in a population of Bryconamericus aff. iheringii (Characidae).

Bryconamericus comprises 56 species distributed into three groups, on the basis of the position and shape of the maxillary teeth: B. exodon, B. microcephalus and B. iheringii groups. Few cytogenetic data are available for this genus, but the diploid number of 52 chromosomes is quite common, although the karyotypic variability is extensive. This study aimed to characterize a population of B. aff. iheringii and thus contribute more cytogenetic information and better understanding of the structure and karyotypic evolution of this genus. We found 6 cytotypes with different NOR patterns: cytotype I showed a karyotype formula of 12m+10sm+16st+14a (FN = 90) and single NORs; cytotype II with 18m+14sm+10st+10a (FN = 94) and cytotype III with 20m+18sm+4st+10a (FN = 94), showing both single and multiple NORs; cytotype IV with 20m+14sm+12st+6a (NF = 98), cytotype V with 22m+18sm+8st+4a (FN = 100) and cytotype VI with 18m+24sm+6st+4a (FN = 100), all with multiple NORs. Cytotype I is the most different in relation to FN and NOR pattern, and can be regarded as belonging to another species of the genus Bryconamericus, living in sympatry in Três Bocas Stream. The remaining cytotypes may have been generated by crosses between them and by pericentric inversions. Meiotic cells were also analyzed and showed that despite the high karyotypic variability, chromosome pairing occurred normally. The great variability found in B. aff. iheringii may be related to a high degree of polymorphism; nevertheless, the possibility of occurrence of more than one species in this location is not ruled out, demonstrating the need for conservation of the Três Bocas Stream.

Physical mapping of 18S rDNA and heterochromatin in species of family Lygaeidae (Hemiptera: Heteroptera).

Analyses conducted using repetitive DNAs have contributed to better understanding the chromosome structure and evolution of several species of insects. There are few data on the organization, localization, and evolutionary behavior of repetitive DNA in the family Lygaeidae, especially in Brazilian species. To elucidate the physical mapping and evolutionary events that involve these sequences, we cytogenetically analyzed three species of Lygaeidae and found 2n (♂) = 18 (16 + XY) for Oncopeltus femoralis; 2n (♂) = 14 (12 + XY) for Ochrimnus sagax; and 2n (♂) = 12 (10 + XY) for Lygaeus peruvianus. Each species showed different quantities of heterochromatin, which also showed variation in their molecular composition by fluorochrome staining. Amplification of the 18S rDNA generated a fragment of approximately 787 bp. The alignment of the consensus sequence with sequences from other species of Heteroptera deposited in the GenBank revealed a similarity of 98% with small differences. Fluorescent in situ hybridization with the 18S rDNA fragment revealed that this ribosomal gene was located in 1 autosomal pair at different positions in the three species. No cytogenetic data are available for these Brazilian species. The basal number and the possible chromosomal changes that occurred among the different species, as well as the evolution of these DNA sequences, are discussed.

Candida albicans and non-C. albicans Candida species: comparison of biofilm production and metabolic activity in biofilms, and putative virulence properties of isolates from hospital environments and infections.

Candida albicans and, more recently, non-C. albicans Candida spp. are considered the most frequent fungi in hospitals. This study analyzed Candida spp. isolates and compared the frequency of different species, that is, C. albicans and non-C. albicans Candida spp., and the origins of isolates, that is, from hospital environments or infections. Yeast virulence factors were evaluated based on biofilm production and metabolic activity. Hemolysin production and the antifungal susceptibility profiles of isolates were also evaluated. Candida spp. were highly prevalent in samples collected from hospital environments, which may provide a reservoir for continuous infections with these yeasts. There were no differences in the biofilm productivity levels and metabolic activities of the environmental and clinical isolates, although the metabolic activities of non-C. albicans Candida spp. biofilms were greater than those of the C. albicans biofilms (p < 0.05). Clinical samples had higher hemolysin production (p < 0.05) and lower susceptibility to fluconazole (p < 0.05). Non-C. albicans Candida spp. predominated in samples collected from hospital environments and infections (p < 0.05). These species had a lower susceptibility to fluconazole and amphotericin B, and their biofilms had higher metabolic activities than those produced by C. albicans, which may explain the increased incidence of fungal infections with these yeasts during recent years.

Remarkable karyotypic homogeneity in a widespread tropical fish species: Hoplosternum littorale (Siluriformes, Callichthyidae).

The first chromosomal data in Hoplosternum littorale from an isolated South American drainage in north-eastern Brazil are presented. All specimens were characterized by a diploid number (2n) of 60 chromosomes divided into three metacentric, one submetacentric and 26 acrocentric pairs; single nucleolar organizer regions (NOR) on the sixth pair; centromeric and interstitial heterochromatin; GC-rich sites on four large acrocentric chromosomes, including the NOR-bearing pair, and 5S ribosomal genes at terminal region on short arms of two acrocentric pairs. These data are invariably similar to previous reports in H. littorale from distant localities throughout South America, which contrasts with the chromosomal diversity of Callichthyidae and reinforces the role of human activities on the dispersal and colonization of this fish.

Sex chromosome differentiation in Belostoma (Insecta: Heteroptera: Belostomatidae).

Belostoma, a genus of the family Belostomatidae, includes species of great ecological importance as biocontrol agents. Few species of these species have been the subject of cytogenetic analyses. Karyotypic evolution in this genus involves agmatoploidy and simploidy; there are also different sex chromosome systems. We examined two Belostoma species (B. dilatatum and B. candidulum) collected from the Paranapanema River Basin (Brazil). Mitotic and meiotic analysis revealed 2n(♂) = 26 + X1X2X3Y for B. dilatatum and 2n(♂) = 14 + XY for B. candidulum; both karyotypes have holokinetic chromosomes. Differences in heterochromatin distribution were also observed between the species, besides variation in the localization of CMA3⁺/DAPI⁻ blocks. The existence of different types of sex chromosome systems in these species was confirmed based on arrangements of the chromosomes in different meiotic stages. We identified a new sex system in B. dilatatum, and make the first cytogenetic report on B. candidulum.