Antifungal effect of (R) and (S)-citronellal enantiomers and their predictive mechanism of action on Candida albicans from voriconazole-resistant onychomycoses / Efeito antifúngico dos enantiômeros (R) e (S)-citronelal e seu mecanismo de ação preditivo sobre Candida albicans de onicomicoses resistentes ao voriconazol

Onychomycosis is the most common disease affecting the nail unit and accounts for at least 50% of all nail diseases. In addition, Candida albicans is responsible for approximately 70% of onychomycoses caused by yeasts. This study investigated the antifungal effect of (R) and (S)-citronellal enantiomers, as well as its predictive mechanism of action on C. albicans from voriconazole-resistant onychomycoses. For this purpose, in vitro broth microdilution and molecular docking techniques were applied in a predictive and complementary manner to the mechanisms of action. The main results of this study indicate that C. albicans was resistant to voriconazole and sensitive to the enantiomers (R) and (S)-citronellal at a dose of 256 and 32 µg/mL respectively. In addition, there was an increase in the minimum inhibitory concentration (MIC) of the enantiomers in the presence of sorbitol and ergosterol, indicating that these molecules possibly affect the integrity of the cell wall and cell membrane of C. albicans. Molecular docking with key biosynthesis proteins and maintenance of the fungal cell wall and plasma membrane demonstrated the possibility of (R) and (S)-citronellal interacting with two important enzymes: 1,3-ß-glucan synthase and lanosterol 14α-demethylase. Therefore, the findings of this study indicate that the (R) and (S)-citronellal enantiomers are fungicidal on C. albicans from onychomycoses and probably these substances cause damage to the cell wall and cell membrane of these micro-organisms possibly by interacting with enzymes in the biosynthesis of these fungal structures.

A onicomicose é a doença mais comum que afeta a unidade ungueal e representa pelo menos 50% de todas as doenças ungueais. Além disso, a Candida albicans é responsável por aproximadamente 70% das onicomicoses causadas por leveduras. Nesse estudo, foi investigado o efeito antifúngico dos enantiômeros (R) e (S)-citronelal, bem como seu mecanismo de ação preditivo sobre C. albicans de onicomicoses resistentes ao voriconazol. Para este propósito, foram aplicadas técnicas in vitro de microdiluição em caldo e docking molecular de forma preditiva e complementar para os mecanismos de ação. Os principais resultados deste estudo indicam que C. albicans foi resistente ao voriconazol e sensível aos enantiômeros (R) e (S)-citronelal na dose de 256 e 32 µg/mL respectivamente. Além disso, houve aumento da concentração inibitória mínima (CIM) dos enantiômeros na presença do sorbitol e do ergosterol, indicando que estas moléculas possivelmente afetem a integridade da parede e da membrana celular de C. albicans. O docking molecular com proteínas chave da biossíntese e manutenção da parede celular e da membrana plasmática fúngica, demonstraram a possibilidade do (R) e (S)-citronelal interagir com duas importantes enzimas: 1,3-ß-glucan sintase e lanosterol 14α-demetilase. Portanto, os achados desse estudo indicam que os enantiômeros (R) e (S)-citronelal são fungicidas sobre C. albicans de onicomicoses e provavelmente essas substâncias causem danos a parede e a membrana celular desses microrganismos possivelmente por interagir com as enzimas da biossíntese destas estruturas fúngicas.

SNPs rs471462296, rs456245081 and rs438495570 in the 5'UTR region of DGAT1 gene in Nelore / SNPs rs471462296, rs456245081 e rs438495570 na região 5'UTR do gene DGAT1 em Nelore

O objetivo desta pesquisa foi avaliar os SNPs rs471462296, rs456245081 e rs438495570 do gene DGAT1 em bovinos Nelore. Foram analisados 109 bovinos. A extração do DNA genômico foi realizada do sangue dos animais, usando-se o kit Ilustra Blood Genomic Prep Mini Spin® (GE Healthcare, UK). A concentração e o grau de pureza do DNA foram determinados por meio de espectrofotômetro (Nanodrop - Thermo Fisher Scientifc, USA). A genotipagem dos SNPs ocorreu mediante o emprego do ensaio Taqman® (Applied Biosystems, USA). Na análise genômica, não foram encontradas alterações nas frequências alélicas e genotípicas (P≥0,05) para os SNPs testados. Dessa forma, a região 5'UTR analisada apresentou-se monomórfica e a variação de SNPs não foi observada, o que limita seu uso como marcadores moleculares para o gene DGAT1 em Nelore.(AU)

Molecular diagnosis of Hepatozoon canis in symptomatic dogs in the city of Goiania, Goiás, Brazil / Diagnóstico molecular de Hepatozoon canis em cães sintomáticos do município de Goiânia, Goiás, Brasil

More than 300 species have been described in the genus Hepatozoon, occurring in different vertebrates. Among these, only Hepatozoon canis and Hepatozoon americanum are seen in dogs. Different methods may be used for laboratory diagnosis. The most common of these is direct parasitological examination of parasite stages in blood smears. The aim of this investigation was to conduct a phylogenetic study on Hepatozoon isolates from symptomatic dogs in the city of Goiânia, Goiás, Brazil. Blood samples were obtained from 40 symptomatic dogs that had been referred to the Veterinary Hospital of the Federal University of Goiás. Among these, only two samples were positive for Hepatozoon spp. using the direct parasitological method. These samples were then subjected to a DNA extraction process and amplification of a fragment of the 18S rRNA by means of PCR. Subsequently, the PCR products from each sample were purified and sequenced. The sequences obtained were then analyzed using the BLASTn algorithm, which identified both sequences of this study as Hepatozoon canis. By applying the Mega4 software, it was confirmed that these isolates of H. canis from dogs in Goiânia are similar to other reference isolates of the same species from other regions of Brazil and worldwide.(AU)

São descritas mais de 300 espécies do gênero Hepatozoon que acometem diferentes vertebrados. Entre estas, apenas Hepatozoon canis e Hepatozoon americanum são descritas em cães. Diferentes métodos podem ser utilizados para o diagnóstico laboratorial. O mais empregado é o exame parasitológico direto do parasito em esfregaços sanguíneos. O objetivo deste trabalho foi realizar um estudo filogenético em Hepatozoon isolados de cães sintomáticos de Goiânia, Goiás. As amostras de sangue foram obtidas de 40 cães sintomáticos encaminhados ao Hospital Veterinário da Universidade Federal de Goiás. Entre essas, duas únicas amostras foram positivas para Hepatozoon spp. pelo método parasitológico direto. Estas amostras foram, então, submetidas ao processo de extração de DNA e de amplificação de um fragmento de 18S rRNA por PCR. Ambas as amostras foram positivas na PCR. Posteriormente, os produtos de PCR de cada amostra foram purificados e sequenciados. As sequências obtidas foram analisadas pelo algoritmo BLASTn, sendo identificadas como Hepatozoon canis. Por meio do software Mega4 foi confirmado que estes isolados de H. canis de cães de Goiânia são semelhantes a outros isolados de referência da mesma espécie de outras regiões do Brasil e do mundo.(AU)

Molecular diagnosis of Hepatozoon canis in symptomatic dogs in the city of Goiania, Goiás, Brazil / Diagnóstico molecular de Hepatozoon canis em cães sintomáticos do município de Goiânia, Goiás, Brasil

More than 300 species have been described in the genus Hepatozoon, occurring in different vertebrates. Among these, only Hepatozoon canis and Hepatozoon americanum are seen in dogs. Different methods may be used for laboratory diagnosis. The most common of these is direct parasitological examination of parasite stages in blood smears. The aim of this investigation was to conduct a phylogenetic study on Hepatozoon isolates from symptomatic dogs in the city of Goiânia, Goiás, Brazil. Blood samples were obtained from 40 symptomatic dogs that had been referred to the Veterinary Hospital of the Federal University of Goiás. Among these, only two samples were positive for Hepatozoon spp. using the direct parasitological method. These samples were then subjected to a DNA extraction process and amplification of a fragment of the 18S rRNA by means of PCR. Subsequently, the PCR products from each sample were purified and sequenced. The sequences obtained were then analyzed using the BLASTn algorithm, which identified both sequences of this study as Hepatozoon canis. By applying the Mega4 software, it was confirmed that these isolates of H. canis from dogs in Goiânia are similar to other reference isolates of the same species from other regions of Brazil and worldwide.(AU)

São descritas mais de 300 espécies do gênero Hepatozoon que acometem diferentes vertebrados. Entre estas, apenas Hepatozoon canis e Hepatozoon americanum são descritas em cães. Diferentes métodos podem ser utilizados para o diagnóstico laboratorial. O mais empregado é o exame parasitológico direto do parasito em esfregaços sanguíneos. O objetivo deste trabalho foi realizar um estudo filogenético em Hepatozoon isolados de cães sintomáticos de Goiânia, Goiás. As amostras de sangue foram obtidas de 40 cães sintomáticos encaminhados ao Hospital Veterinário da Universidade Federal de Goiás. Entre essas, duas únicas amostras foram positivas para Hepatozoon spp. pelo método parasitológico direto. Estas amostras foram, então, submetidas ao processo de extração de DNA e de amplificação de um fragmento de 18S rRNA por PCR. Ambas as amostras foram positivas na PCR. Posteriormente, os produtos de PCR de cada amostra foram purificados e sequenciados. As sequências obtidas foram analisadas pelo algoritmo BLASTn, sendo identificadas como Hepatozoon canis. Por meio do software Mega4 foi confirmado que estes isolados de H. canis de cães de Goiânia são semelhantes a outros isolados de referência da mesma espécie de outras regiões do Brasil e do mundo.(AU)

Detection of Salmonella spp. by Conventional Bacteriology and by Quantitative Polymerase-Chain Reaction in Commercial Egg Structures

Conventional bacteriology techniques and quantitative polymerase-chain reaction (qPCR) were applied to the eggshell, albumen, and yolk of washed and unwashed commercial white and brown eggs, to detect Salmonella spp. Pooled samples of eggshells, albumen, and yolk of white and brown eggs were collected at the poultry house and at the egg-storage room. Salmonella spp. was detected by conventional bacteriology in 5.4% (21/387) of analyzed samples and in 16% (68/387) by qPCR. In the 114 unwashed white eggs samples of eggshell, albumen and yolk, the bacterium was identified in 2.6% of the eggs (3/114) by conventional bacteriology and in 13.2% (15/114) by qPCR. In the 90 samples of washed eggs, 6.7% (6/90) were contaminated as detected by conventional bacteriology and 10.0% (9/90) by qPCR. In the 81 samples of unwashed brown eggs, Salmonella spp. was detected in 6.1% of the eggs (5/81) by conventional bacteriology and 27.2% (22/81) by qPCR. In the 102 samples of brown washed eggs, 6.9% (7/102) where positive by conventional bacteriology and 35.3% (16/102) by qPCR. All samples detected as positive by conventional bacteriology were also positive by qPCR. Salmonella Agona represented 18.2% (4/22) of identified serovars, Salmonella enterica subs. enterica O: 4.5 18.2% (4/22), Salmonella Schwarzengrund 18.2% (4/22), Salmonella Cerro 13.6% (3/22), Salmonella Anatum 13.6% (3/22), Salmonella Enteritidis 9.1% (2/22), Salmonella Johannesburg 4.5% (1/22), and Salmonella Corvallis 4.5% (1/22). The qPCR method provided better detection of Salmonella spp. in commercial eggs than conventional bacteriology. The conventional egg washing and disinfection procedures are not efficient to eliminate Salmonella.

Detection of Salmonella spp. by Conventional Bacteriology and by Quantitative Polymerase-Chain Reaction in Commercial Egg Structures

Conventional bacteriology techniques and quantitative polymerase-chain reaction (qPCR) were applied to the eggshell, albumen, and yolk of washed and unwashed commercial white and brown eggs, to detect Salmonella spp. Pooled samples of eggshells, albumen, and yolk of white and brown eggs were collected at the poultry house and at the egg-storage room. Salmonella spp. was detected by conventional bacteriology in 5.4% (21/387) of analyzed samples and in 16% (68/387) by qPCR. In the 114 unwashed white eggs samples of eggshell, albumen and yolk, the bacterium was identified in 2.6% of the eggs (3/114) by conventional bacteriology and in 13.2% (15/114) by qPCR. In the 90 samples of washed eggs, 6.7% (6/90) were contaminated as detected by conventional bacteriology and 10.0% (9/90) by qPCR. In the 81 samples of unwashed brown eggs, Salmonella spp. was detected in 6.1% of the eggs (5/81) by conventional bacteriology and 27.2% (22/81) by qPCR. In the 102 samples of brown washed eggs, 6.9% (7/102) where positive by conventional bacteriology and 35.3% (16/102) by qPCR. All samples detected as positive by conventional bacteriology were also positive by qPCR. Salmonella Agona represented 18.2% (4/22) of identified serovars, Salmonella enterica subs. enterica O: 4.5 18.2% (4/22), Salmonella Schwarzengrund 18.2% (4/22), Salmonella Cerro 13.6% (3/22), Salmonella Anatum 13.6% (3/22), Salmonella Enteritidis 9.1% (2/22), Salmonella Johannesburg 4.5% (1/22), and Salmonella Corvallis 4.5% (1/22). The qPCR method provided better detection of Salmonella spp. in commercial eggs than conventional bacteriology. The conventional egg washing and disinfection procedures are not efficient to eliminate Salmonella.(AU)

Expressão gênica de adipocinas em ovelhas alimentadas com resíduos da indústria do biodiesel da mamona / Adipokine gene expression in ewes fed with residues from the biodiesel castor industry

A expressão de RNAm para leptina, receptor de leptina (obRb), adiponectina, receptor de adiponectina (AdipoR1) e resistina foi avaliada por meio da técnica de PCR em tempo real, em tecidos ovariano, hipofisário, adiposo do omento e da região perirrenal, em ovelhas alimentadas sem farelo de mamona ou com farelo de mamona detoxificada durante 14 meses. O tipo de dieta não afetou os níveis de RNAm para leptina, obRb, adiponectina, AdipoR1 e resistina nos diferentes tecidos avaliados (P>0,05). Nos tecidos ovariano e hipofisário, não foi verificada a expressão da adiponecina e da resistina, respectivamente. Como consequência, pode-se concluir que o farelo de mamona detoxificada pode ser utilizado como fonte proteica na dieta de ovelhas, sem afetar a expressão do gene resistina e dos genes leptina e adiponectina, bem como de seus receptores.(AU)

The expression of leptin, leptin receptor (obRb), adiponectin, adiponectin receptor (AdipoR1) and resistin was assessed by real-time PCR technique in ovarian, pituitary, and the omental adipose perirenal tissue in sheep feed without castor meal or with detoxified castor meal. The type of diet did not affect mRNA levels for leptin, obRb, adiponectin, resistin AdipoR1 evaluated in different tissues (P>0.05). However, in pituitary and ovarian tissues there was no expression of resistin and adiponectin, respectively. The detoxified castor meal can be used in sheep diets as alternative food protein without affecting the expression of leptin and adponectin as well as their receptors and resistin.(AU)

Ciclo reprodutivo de Parauchenipterus striatulus (Pisces - Auchenipteridae) na represa de Ribeirão das Lajes - RJ

The objective of this work was to evaluate aspects related to the reproductive cycle of the armored catfish Parauchenipterus striatulus in Ribeirão das Lajes Reservoir, state of Rio de Janeiro, Brazil. Gonadal stage, morphology, variations in the gonadosomatic index, weight:length relationship, condition factor and gonadal condition were described. Fish sampling was carried out bi-monthly, between April 1996 and May 1997, using gill nets 100m long, 4m wide, and 25-65mm of mesh sizes gill nets. Three hundred and thirty-nine fishes, corresponding to 40% of the total number, were capture. Five maturation stages were determined by macroscopic analysis of both sexes: immature, mature I, mature II, ripe and spawned (females) or empty (males). The gonadosomatic index was higher from October to March for both sexes, suggesting an ample spawning period, while the condition factor was higher from February to June, showing an inverse relationship between these two characteristics. Weight:length relationship for males was W = 0.0095L3.0862 and for females W = 0.0116L3.0126, with no significant differences between sexes in the allometric coefficient.

Este trabalho teve como objetivo avaliar o ciclo reprodutivo do bagre de água doce Parauchenipterus striatulus, na represa de Ribeirão das Lajes, Rio de Janeiro. Foram descritos aspectos morfológicos dos estádios de maturação gonadal bem como variações no índice gonadossomático, relação peso:comprimento, fator de condição e condição gonadal. A amostragem dos peixes foi feita bimestralmente entre abril de 1996 e maio de 1997 com o uso de redes de espera de 100m de comprimento, 4m de altura e com malha entre 25 e 65mm de distância entrenós. Os 339 indivíduos capturados ao longo do período de amostragem corresponderam a 40% do total de peixes. Cinco estádios de maturação gonadal foram determinados por meio de análise macroscópica para ambos os sexos: imaturo, maturação I, maturação II, maduro, desovado (fêmeas) e esvaziado (machos). O índice gonadossomático foi mais elevado entre outubro e março para ambos os sexos, sugerindo amplo período de desova enquanto o fator de condição atingiu seu máximo entre fevereiro e junho, mostrando uma relação inversa entre essas duas características. A relação peso-comprimento para machos foi de W= 0,0095L3,0862 e fêmeas de W= 0,0116L3,0126, não tendo sido encontradas diferenças significativas entre os coeficientes de alometria para ambos os sexos.