RESUMO

In the current context of emerging drug-resistant fungal pathogens such as Candida albicans and Candida parapsilosis, discovery of new antifungal agents is an urgent matter. This research aimed to evaluate the antifungal potential of 2-chloro-N-phenylacetamide against fluconazole-resistant clinical strains of C. albicans and C. parapsilosis. The antifungal activity of 2-chloro-N-phenylacetamide was evaluated in vitro by the determination of the minimum inhibitory concentration (MIC), minimum fungicidal concentration (MFC), inhibition of biofilm formation and its rupture, sorbitol and ergosterol assays, and association between this molecule and common antifungal drugs, amphotericin B and fluconazole. The test product inhibited all strains of C. albicans and C. parapsilosis, with a MIC ranging from 128 to 256 µg.mL-1, and a MFC of 512-1,024 µg.mL-1. It also inhibited up to 92% of biofilm formation and rupture of up to 87% of preformed biofilm. 2-chloro-N-phenylacetamide did not promote antifungal activity through binding to cellular membrane ergosterol nor it damages the fungal cell wall. Antagonism was observed when combining this substance with amphotericin B and fluconazole. The substance exhibited significant antifungal activity by inhibiting both planktonic cells and biofilm of fluconazole-resistant strains. Its combination with other antifungals should be avoided and its mechanism of action remains to be established.

No atual contexto de patógenos fúngicos resistentes emergentes tais como Candida albicans e Candida parapsilosis, a descoberta de novos agentes antifúngicos é uma questão urgente. Esta pesquisa teve como objetivo avaliar o potencial antifúngico da 2-cloro-N-fenilacetamida contra cepas clínicas de C. albicans e C. parapsilosis resistentes a fluconazol. A atividade antifúngica da substância foi avaliada in vitro através da determinação da concentração inibitória mínima (CIM), concentração fungicida mínima (CFM), ruptura e inibição da formação de biofilme, ensaios de sorbitol e ergosterol, e associação entre esta molécula e antifúngicos comuns, anfotericina B e fluconazol. O produto teste inibiu todas as cepas de C. albicans e C. parapsilosis, com uma CIM variando de 128 a 256 µg.mL-1, e uma CFM de 512-1,024 µg.mL-1. Também inibiu até 92% da formação de biofilme e causou a ruptura de até 87% de biofilme pré-formado. A 2-cloro-N-fenilacetamida não promoveu atividade antifúngica pela ligação ao ergosterol da membrana celular fúngica, tampouco danificou a parede celular. Antagonismo foi observado ao combinar esta substância com anfotericina B e fluconazol. A substância exibiu atividade antifúngica significativa ao inibir tanto as células planctônicas quanto o biofilme das cepas resistentes ao fluconazol. Sua combinação com outros antifúngicos deve ser evitada e seu mecanismo de ação deve ser estabelecido.

Assuntos

RESUMO

Abstract In the current context of emerging drug-resistant fungal pathogens such as Candida albicans and Candida parapsilosis, discovery of new antifungal agents is an urgent matter. This research aimed to evaluate the antifungal potential of 2-chloro-N-phenylacetamide against fluconazole-resistant clinical strains of C. albicans and C. parapsilosis. The antifungal activity of 2-chloro-N-phenylacetamide was evaluated in vitro by the determination of the minimum inhibitory concentration (MIC), minimum fungicidal concentration (MFC), inhibition of biofilm formation and its rupture, sorbitol and ergosterol assays, and association between this molecule and common antifungal drugs, amphotericin B and fluconazole. The test product inhibited all strains of C. albicans and C. parapsilosis, with a MIC ranging from 128 to 256 µg.mL-1, and a MFC of 512-1,024 µg.mL-1. It also inhibited up to 92% of biofilm formation and rupture of up to 87% of preformed biofilm. 2-chloro-N-phenylacetamide did not promote antifungal activity through binding to cellular membrane ergosterol nor it damages the fungal cell wall. Antagonism was observed when combining this substance with amphotericin B and fluconazole. The substance exhibited significant antifungal activity by inhibiting both planktonic cells and biofilm of fluconazole-resistant strains. Its combination with other antifungals should be avoided and its mechanism of action remains to be established.

Resumo No atual contexto de patógenos fúngicos resistentes emergentes tais como Candida albicans e Candida parapsilosis, a descoberta de novos agentes antifúngicos é uma questão urgente. Esta pesquisa teve como objetivo avaliar o potencial antifúngico da 2-cloro-N-fenilacetamida contra cepas clínicas de C. albicans e C. parapsilosis resistentes a fluconazol. A atividade antifúngica da substância foi avaliada in vitro através da determinação da concentração inibitória mínima (CIM), concentração fungicida mínima (CFM), ruptura e inibição da formação de biofilme, ensaios de sorbitol e ergosterol, e associação entre esta molécula e antifúngicos comuns, anfotericina B e fluconazol. O produto teste inibiu todas as cepas de C. albicans e C. parapsilosis, com uma CIM variando de 128 a 256 µg.mL-1, e uma CFM de 512-1,024 µg.mL-1. Também inibiu até 92% da formação de biofilme e causou a ruptura de até 87% de biofilme pré-formado. A 2-cloro-N-fenilacetamida não promoveu atividade antifúngica pela ligação ao ergosterol da membrana celular fúngica, tampouco danificou a parede celular. Antagonismo foi observado ao combinar esta substância com anfotericina B e fluconazol. A substância exibiu atividade antifúngica significativa ao inibir tanto as células planctônicas quanto o biofilme das cepas resistentes ao fluconazol. Sua combinação com outros antifúngicos deve ser evitada e seu mecanismo de ação deve ser estabelecido.

RESUMO

In the current context of emerging drug-resistant fungal pathogens such as Candida albicans and Candida parapsilosis, discovery of new antifungal agents is an urgent matter. This research aimed to evaluate the antifungal potential of 2-chloro-N-phenylacetamide against fluconazole-resistant clinical strains of C. albicans and C. parapsilosis. The antifungal activity of 2-chloro-N-phenylacetamide was evaluated in vitro by the determination of the minimum inhibitory concentration (MIC), minimum fungicidal concentration (MFC), inhibition of biofilm formation and its rupture, sorbitol and ergosterol assays, and association between this molecule and common antifungal drugs, amphotericin B and fluconazole. The test product inhibited all strains of C. albicans and C. parapsilosis, with a MIC ranging from 128 to 256 µg.mL-1, and a MFC of 512-1,024 µg.mL-1. It also inhibited up to 92% of biofilm formation and rupture of up to 87% of preformed biofilm. 2-chloro-N-phenylacetamide did not promote antifungal activity through binding to cellular membrane ergosterol nor it damages the fungal cell wall. Antagonism was observed when combining this substance with amphotericin B and fluconazole. The substance exhibited significant antifungal activity by inhibiting both planktonic cells and biofilm of fluconazole-resistant strains. Its combination with other antifungals should be avoided and its mechanism of action remains to be established.

Assuntos

RESUMO

The objective was to evaluate the effect of dietary 25-hydroxycholecalciferol (25(OH)D3) or vitamin D3 (VitD3) and different total calcium (Ca) levels on the performance, carcass characteristics, blood, enzymatic, and bone biochemistry of broilers reared under heat stress between 1 and 42 days of age. A total of 504 male, Cobb 500, broiler chickens were distributed in a completely randomized design in a 2 × 4 factorial arrangement (VitD3 or 25(OH)D3 × four Ca levels (100, 90, 80 and 70% of the recommendations of Rostagno et al. (2011)), eight treatments, seven replicates and nine broilers per cage. Feed intake and feed conversion ratio did not (p>0.05) vary when levels of Ca were reduced and vitamin D3 sources were supplemented in the diets from 1 to 21 days for broilers chickens. 25 (OH)D3 increased weight gain results (p<0.05). From 1 to 42 d, no differences (p>0.05) were observed on performance, carcass yields and meat quality, bone deposition of Ca and P, and alkaline phosphatase concentration. Higher serum (p<0.05) concentrations of Ca and P were found in broilers fed with 25(OH)D3. The replacement of VitD3 with 25(OH)D3 and the Ca reduction of 30% in diets did not negatively affect performance, carcass characteristics, and Ca and P deposition in the tibia of broilers at 42 days of age, under heat stress.(AU)

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In modern pig farming, the search for systems that promote environmental quality and welfare is an important issue. In this sense, the present study evaluated the effects of environmental enrichment on the performance and behavior of piglets. In a completely randomized block design, 32 piglets (7.43kg mean weight), weaned at 28 days of age, were distributed into four treatments (control without enrichment, environmental enrichment with wood shavings as bedding; environmental enrichment with hanging toys and environmental enrichment with wood shavings + hanging toys). Four repetitions were performed for each treatment, with two animals per experimental unit. The instant scan sampling technique was used to record the behavior of each piglet for 20h. Animals receiving environmental enrichment with wood shavings + hanging toys were heavier at 70 days (P=0.02), exhibited higher total and daily weight gain (P=0.04 and P=0.02, respectively) and better feed conversion (P=0.02). Environmental enrichment increased the interaction of animals with the environment, allowing them to exhibit natural behaviors.

Na suinocultura moderna, a busca de sistemas que promovam a qualidade e o bem-estar do meio ambiente é uma questão importante. Nesse sentido, o presente estudo avaliou os efeitos do enriquecimento ambiental sobre o desempenho e o comportamento de leitões. Em um delineamento em blocos ao acaso, 32 leitões (7,43kg de peso médio), desmamados aos 28 dias de idade, foram distribuídos em quatro tratamentos (controle sem enriquecimento, enriquecimento ambiental com cama de maravalha, enriquecimento com móbiles e cama + móbiles). Quatro repetições foram realizadas para cada tratamento, com dois animais por unidade experimental. A técnica de amostragem de varredura instantânea foi usada para registrar o comportamento de cada leitão, totalizando 20h. Os animais que receberam enriquecimento ambiental com cama + móbiles foram mais pesados aos 70 dias (P = 0,02), apresentaram maior ganho de peso total e diário (P = 0,04 e P = 0,02, respectivamente) e melhor conversão alimentar (P = 0,02). O enriquecimento ambiental aumentou a interação dos animais com o meio ambiente, o que lhes permite exibir comportamentos típicos.

Assuntos

RESUMO

In modern pig farming, the search for systems that promote environmental quality and welfare is an important issue. In this sense, the present study evaluated the effects of environmental enrichment on the performance and behavior of piglets. In a completely randomized block design, 32 piglets (7.43kg mean weight), weaned at 28 days of age, were distributed into four treatments (control without enrichment, environmental enrichment with wood shavings as bedding; environmental enrichment with hanging toys and environmental enrichment with wood shavings + hanging toys). Four repetitions were performed for each treatment, with two animals per experimental unit. The instant scan sampling technique was used to record the behavior of each piglet for 20h. Animals receiving environmental enrichment with wood shavings + hanging toys were heavier at 70 days (P=0.02), exhibited higher total and daily weight gain (P=0.04 and P=0.02, respectively) and better feed conversion (P=0.02). Environmental enrichment increased the interaction of animals with the environment, allowing them to exhibit natural behaviors.(AU)

Na suinocultura moderna, a busca de sistemas que promovam a qualidade e o bem-estar do meio ambiente é uma questão importante. Nesse sentido, o presente estudo avaliou os efeitos do enriquecimento ambiental sobre o desempenho e o comportamento de leitões. Em um delineamento em blocos ao acaso, 32 leitões (7,43kg de peso médio), desmamados aos 28 dias de idade, foram distribuídos em quatro tratamentos (controle sem enriquecimento, enriquecimento ambiental com cama de maravalha, enriquecimento com móbiles e cama + móbiles). Quatro repetições foram realizadas para cada tratamento, com dois animais por unidade experimental. A técnica de amostragem de varredura instantânea foi usada para registrar o comportamento de cada leitão, totalizando 20h. Os animais que receberam enriquecimento ambiental com cama + móbiles foram mais pesados aos 70 dias (P = 0,02), apresentaram maior ganho de peso total e diário (P = 0,04 e P = 0,02, respectivamente) e melhor conversão alimentar (P = 0,02). O enriquecimento ambiental aumentou a interação dos animais com o meio ambiente, o que lhes permite exibir comportamentos típicos.(AU)

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Low-level laser therapy treatment (LLLT) is widely used in rehabilitation clinics with the aim of accelerating the process of tissue repair; however, the molecular bases of the effect of LLLT have not been fully established. The aim of the present study was to evaluate the influence of the exposure of different doses of LLLT on the expression of collagen genes type I alpha 1 (COL1α1) and vascular endothelial growth factor (VEGF) in the fibroblast cells of mice (L929) cultivated in vitro. Fibroblast cells were irradiated with a Gallium-Arsenide laser (904 nm) every 24 h for 2 consecutive days, stored in an oven at 37 °C, with 5% CO2 and divided into 3 groups: G1-control group, G2-irradiated at 2 J/cm(2), and G3-irradiated at 3 J/cm(2). After irradiation, the total RNA was extracted and used in the complementary DNA (cDNA) synthesis. The gene expression was analyzed by real-time polymerase chain reaction. The cells irradiated in G2 exhibited a statistically significant growth of 1.78 in the expression of the messenger RNA (mRNA) of the COL1α1 gene (p = 0.036) in comparison with G1 and G3. As for the VEGF gene, an increase in expression was observed in the two irradiated groups in comparison with the control group. There was an increase in expression in G2 of 2.054 and G3 of 2.562 (p = 0.037) for this gene. LLLT (904 nm) had an influence on the expression of the genes COL1α1 (2 J/cm(2)) and VEGF (2 e 3 J/cm(2)) in a culture of the fibroblast cells of mice.

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The purpose of this study was to evaluate the effect of temperature and different levels of available phosphorus (aP) on the expression of nine genes encoding electron transport chain proteins in the Longissimus dorsi (LD) muscle of pigs. Two trials were carried out using 48 high-lean growth pigs from two different growth phases: from 15 to 30 kg (phase 1) and from 30 to 60 kg (phase 2). Pigs from growth phase 1 were fed with three different levels of dietary aP (0.107%, 0.321% or 0.535%) and submitted either to a thermoneutral (24°C and RH at 76%) or to a heat stress (34°C and RH at 70%) environment. Pigs from growth phase 2 were fed with three different levels of dietary aP (0.116%, 0.306% or 0.496%) and submitted either to a thermoneutral (22ºC and RH at 77%) or to a heat stress (32ºC and RH at 73%) environment. Heat stress decreased (P<0.001) average daily feed intake at both growth phases. At 24°C, pigs in phase 1 fed the 0.321% aP diet had greater average daily gain and feed conversion (P<0.05) than those fed the 0.107% or 0.535% while, at 34°C pigs fed the 0.535% aP had the best performance (P<0.05). Pigs from phase 2 fed the 0.306% aP had best performance in both thermal environments. Gene expression profile was analyzed by quantitative real-time polymerase chain reaction. Irrespective of growing phase, the expression of six genes was lower (P<0.05) at high temperature than at thermoneutrality. The lower expression of these genes under high temperatures evidences the effects of heat stress by decreasing oxidative metabolism, through adaptive physiological mechanisms in order to reduce heat production. In pigs from phase 1, six genes were differentially expressed across aP levels (P<0.05) in the thermoneutral and one gene in the heat stress. In pigs from phase 2, two genes were differentially expressed across aP levels (P<0.05) in both thermal environments. These data revealed strong evidence that phosphorus and thermal environments are key factors to regulate oxidative phosphorylation with direct implications on animal performance.

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Avaliou-se a suplementação de dietas com fósforo disponível (Pd) para suínos selecionados para deposição de carne, utilizando-se 100 leitões com média de peso inicial de 15,03±0,33kg, distribuídos em delineamento de blocos ao acaso, com cinco tratamentos, cinco repetições e quatro animais por unidade experimental. Os tratamentos foram dieta basal sem suplementação de fosfato bicálcico e outras quatro dietas obtidas pela suplementação da dieta basal com fosfato bicálcico, resultando em cinco concentrações de Pd, 0,106; 0,221; 0,310; 0,406; e 0,493%. As porcentagens de Pd influenciaram o consumo diário de ração, que aumentou de forma linear. O ganho de peso diário aumentou e a conversão alimentar melhorou de forma quadrática até a suplementação máxima de 0,450 e 0,390% de Pd, respectivamente. Não houve efeito da suplementação das dietas sobre a resistência óssea. As porcentagens de Pd influenciaram de forma quadrática os teores de cálcio e de fósforo nos ossos, que aumentaram até a adição máxima de 0,388 e 0,369% de Pd, respectivamente. Não houve efeito da porcentagem de Pd sobre o teor de cinza óssea. Suínos dos 15 aos 30kg exigem maior concentração de Pd para máximo ganho de peso (0,450% ou 5,33g/dia) e melhor conversão alimentar (0,390% ou 4,52g/dia) em relação às recomendações atuais.

The supplementation of diets with available phosphorus (aP) for pigs selected for meat deposition was evaluated, using 100 commercial hybrid piglets, 50 castrated males and 50 females, with average initial weight of 15.03±0.33kg allotted in a randomized block design with five treatments, five replicates, and four pigs, 2 males and 2 females per experimental unit. The treatments were a basal diet and other four diets obtained through basal diet supplementation with dicalcium phosphate to obtain five aP concentrations, 0.106, 0.221, 0.310, 0,406, and 0.493%. The percentages of aP influenced daily feed intake, which increased linearly. Daily weight gain increased and feed conversion improved quadratically up to 0.450 and 0.390% maximum responses. There was no effect of diet supplementation on bone strength. The percentages of aP influenced the calcium and phosphorus contents in the bones, which increased quadratically up to 0.388 and 0.369% maximum responses, respectively. There was no effect of aP percentage in the bone ashe content. Pigs from 15 to 30kg require greater amount of aP in the diet for maximal weight gain (0.450% or 5.33g/d) and greatest feed conversion (0.390% or 4.52g/d) compared to current requirements.

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Electroejaculation is a technique that can be used to collect semen from canines, but its use with this group of animals is restricted by low success rate and low semen quality. Here, we evaluated whether pharmacological and sexual sensory stimuli, which may affect ejaculation, can increase electroejaculation efficiency and improve ejaculate quality. We worked with 20 dogs of mixed breed weighing between 5.3 and 22.2 kg, divided into two groups. Both groups were exposed to a spayed female for 10 min, but in the second group, the same spayed female had her vagina impregnated with methyl 4-hydroxybenzoate synthetic pheromone for 10 min and after receiving dinoprost tromethamine IM, 0.1 mg/kg. After stimulation, all dogs were chemically restrained with ketamine, 8 mg/kg, IM; and xylazine, 1 mg/kg, IM, and subjected to electroejaculation protocol. We obtained 100% of antegrade ejaculate in treatments when the spayed female had her vagina impregnated with pheromone and 80% when she did not. Sperm motility was significantly different (p < 0.05) between controls and the test group (10.1 ± 4.5 and 43.0 ± 8.3, respectively). We concluded that the adopted electroejaculation protocol was efficient and that the PGF2α associated with sexual sensory stimulation can improve semen quality in dogs undergoing the procedure.

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