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1.
Int J Mol Sci ; 25(16)2024 Aug 15.
Artículo en Inglés | MEDLINE | ID: mdl-39201576

RESUMEN

This study aimed to evaluate the effects of supplementation with ethanolic and aqueous extracts from the bark of the stem of Guazuma ulmifolia in mice submitted to a high-fat diet as well as to evaluate the chemical composition of these extracts. The chemical composition and antioxidant potential was evaluated in aqueous and ethanolic extracts of the stem bark. The in vivo test consisted of evaluating the effects of the aqueous and ethanolic extracts of the stem bark on C57BL/6 mice receiving a high-fat diet. The animals were evaluated for weight gain, feed consumption, visceral adiposity, serum, and inflammatory and hormonal parameters. The results of the chemical analyses corroborate those obtained by the literature, which reported gallocatechin, epigallocatechin and epigallocatechin gallate. Compared with the ethanolic extract, the aqueous extract showed greater antioxidant capacity. Both extracts resulted in lower feed consumption in the animals, but they did not influence weight gain or visceral adiposity and resulted in varied changes in the lipid profile. In addition, they did not influence glucose tolerance, insulin sensitivity, or fasting blood glucose. Furthermore, the leptin levels increased, which may have contributed to satiety, but this was shown to have a negative impact on other inflammatory and hormonal parameters. Therefore, under the conditions of this study, the biologically active compounds present in the plant species Guazuma ulmifolia were not able to contribute to the treatment of metabolic changes related to the consumption of a high-fat diet.


Asunto(s)
Antioxidantes , Dieta Alta en Grasa , Enfermedades Metabólicas , Ratones Endogámicos C57BL , Corteza de la Planta , Extractos Vegetales , Animales , Extractos Vegetales/farmacología , Extractos Vegetales/química , Dieta Alta en Grasa/efectos adversos , Corteza de la Planta/química , Ratones , Enfermedades Metabólicas/tratamiento farmacológico , Enfermedades Metabólicas/prevención & control , Enfermedades Metabólicas/etiología , Antioxidantes/farmacología , Masculino , Apocynaceae/química , Tallos de la Planta/química , Glucemia/efectos de los fármacos
2.
Foods ; 13(14)2024 Jul 20.
Artículo en Inglés | MEDLINE | ID: mdl-39063367

RESUMEN

Grape seeds are rich in bioactive substances, including polyphenols, terpenoids, and phytosterols. Linseed (Linum usitatissimum L.) boasts a high concentration of polyunsaturated fatty acids (PUFAs), lignans, phytoestrogens, and soluble fibers, all contributing to its therapeutic potential. In this study, we pioneered the formulation of an oil blend (GL) combining grape seed oil (G) and golden linseed oil (GL) in equal volumes (1:1 (v/v)) and we evaluated in terms of the nutritional, physical, and chemical properties and their influence in an in vivo experimental model. We analyzed the oils by performing physical-chemical analyses, examining the oxidative stability using Rancimat; conducting thermal analyses via thermogravimetry/derivative thermogravimetry (TG/DTG) and differential scanning calorimetry (DSC), performing optical UV-vis absorption analyses; examining the fluorescence emission-excitation matrix, total carotenoids, and color, and conducting metabolic assessments in an in vivo experimental trial. The fatty acid profile presented a higher fraction of linoleic acid (C18:2) in G and GL and alpha-linolenic acid (C18:3) in L. The acidity and peroxide indices were within the recommended ranges. The TG/DTG, DSC, and Rancimat analyses revealed similar behaviors, and the optical analyses revealed color variations caused by carotenoid contents in L and GL. In the in vivo trial, G (G2: 2000 mg/kg/day) promoted lower total consumption, and the blend (GL: 2000 mg/kg/day) group exhibited less weight gain per gram of consumed food. The group with G supplementation (G2: 2000 mg/kg/day) and GL had the highest levels of HDL-c. The group with L supplementation (L2: 2000 mg/kg/day) had the lowest total cholesterol level. The L2, G1 (1000 mg/kg/day), and G2 groups exhibited the lowest MCP-1 and TNF-α values. Additionally, the lowest adipocyte areas occurred in G and GL. Our results suggest that this combination is of high quality for consumption and can influence lipid profiles, markers of inflammation, and antioxidant status.

3.
Molecules ; 28(23)2023 Nov 28.
Artículo en Inglés | MEDLINE | ID: mdl-38067541

RESUMEN

Vitis vinifera L. (grapevine) is a perennial plant of the Vitaceae family that is widely used to produce grapes and wines. Grape seed oil is rich in fatty acids such as linoleic acid (65-75%), vitamin E (50 mg), and phytosterols in addition to phenolic compounds, such as catechins (414 mg), epicatechins (130.4 mg), and gallic acid (77 µg), shows promise as a nutritional compound and is outstanding as a therapeutic substance with active properties for health, detected mainly by in vitro studies, as well as some in vivo studies. The benefits of consuming this oil include modulating the expression of antioxidant enzymes, anti-atherosclerotic and anti-inflammatory effects, and protection against oxidative cell damage and some types of cancer. However, experimental findings confirm that therapeutic functions remain scarce; thus, more studies are needed to determine the mechanisms of action involved in the indicated therapeutic qualities.


Asunto(s)
Vitis , Humanos , Antioxidantes/farmacología , Estrés Oxidativo , Suplementos Dietéticos , Obesidad , Inflamación/tratamiento farmacológico , Aceites de Plantas/farmacología , Frutas
4.
Nutrients ; 15(6)2023 Mar 07.
Artículo en Inglés | MEDLINE | ID: mdl-36986038

RESUMEN

The study of medicinal plants, such as the genus Garcinia (Clusiaceae), in the treatment of non-communicable chronic diseases has aroused the interest of researchers. However, there are no studies in the literature that have investigated the effects of Garcinia gardneriana in experimental models of obesity for possible metabolic alterations. Swiss mice receiving a high-fat diet were supplemented with aqueous or ethanolic extract of G. gardneriana at doses of 200 or 400 mg/kg/day. It was found that there was a reduction in food consumption in experimental groups compared with the control groups, and the group supplemented with aqueous extract at a dose of 200 mg/kg/daydisplayed a reduction in weight. The results showed an increase in the values of high density lipoprotein (HDL-c), total cholesterol, triglycerides and fasting blood glucose. G. gardneriana did not protect against insulin resistance, and caused in an increase in monocyte chemoattractant protein-1 (MCP-1) concentrations and a reduction in interleukin 10 (IL-10). In addition, hepatic steatosis and microvesicular steatosis were indicated. It was revealed that, under the experimental conditions in the study, G. gardneriana did not prevent weight gain or comorbidities; that is, a different behavior was obtained from that described in the literature with regard to the medicinal potential of the Garcinia species, which is probably related to the phytochemical properties.


Asunto(s)
Hígado Graso , Garcinia , Plantas Medicinales , Ratones , Animales , Garcinia/química , Extractos Vegetales/farmacología , Etanol , Agua , Dieta Alta en Grasa/efectos adversos
5.
Nutrients ; 14(6)2022 Mar 10.
Artículo en Inglés | MEDLINE | ID: mdl-35334818

RESUMEN

Different fatty acid proportions produce potential inflammatory and metabolic changes in organisms. However, the evidence for how each fatty acid mediates the metabolic pathway, and its lipid stability remains controversial. To resolve this controversy, the present study investigated the metabolic effects of cold-pressed linseed (LG), coconut (CG), and baru (BG) oils in comparison to those of soybean oil (SG) in mice, in terms of their oil characterization and stability. The quality analysis showed less oxidative behavior among PUFA-rich oils (SO, BO, and LO, with induction periods lower than 2 h compared to 39.8 h for CG), besides the high contents of tocopherols and carotenoids in SG and LG. In the experimental study, CG presented higher triglyceride (257.93 ± 72.30) and VLDL-cholesterol levels (51.59 ± 14.46, p < 0.05), while LG reduced LDL levels (59.29 ± 7.56, p < 0.05) when compared to SG (183.14 ± 22.06, 36.63 ± 4.41 and 131.63 ± 29.0, respectively). For visceral fats, the adiposity index was lower for BG (7.32 ± 3.13) and CG (9.58 ± 1.02, p < 0.05) in relation to SG (12.53 ± 2.80), and for leukocyte recruitment, CG presented lower polymorphonuclear (PMN) (p < 0.0001) and mononuclear (MN) (p < 0.05) cell infiltration, demonstrating anti-inflammatory potential. In NMR-based metabolomics, although CG presented higher values for the glucose, lactate, and LDL/VLDL ratio, this group also evidenced high levels of choline, a lipotropic metabolite. Our study emphasized the controversies of saturated fatty acids, which impair serum lipids, while alfa-linolenic acid presented cardioprotective effects. However, coconut oil also has a positive immunomodulatory pathway and was found to reduce visceral bodyfat in mice. Therefore, for future applications, we suggest a combination of lauric and al-fa-linolenic acid sources, which are present in coconut and linseed oil, respectively. This combination could be less obesogenic and inflammatory and exert cardioprotective action.


Asunto(s)
Lino , Aceite de Linaza , Animales , Aceite de Coco , Cocos , Leucocitos , Metabolómica , Ratones
6.
J Med Food ; 24(8): 841-851, 2021 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-34342510

RESUMEN

Obesity is a relevant health hazard characterized as a chronic noncommunicable disease, with severe comorbidities that cause mortality worldwide. Acrocomia aculeata is a Brazilian palm with edible fruits. Its pulp contains fibers, monounsaturated fatty acids (MUFAs), such as oleic acid and carotenoids. In this context, our study aimed to elucidate the protective effect of the lyophilized A. aculeata pulp added at the rates of 1%, 2%, and 4% to a high-fat (HF) diet (rich in saturated fats and cholesterol), for 90 days, in mice. The treatment with 4% pulp induced a significant increase in the biochemical parameters of serum cholesterol HDL-C (high-density lipoprotein) compared with the control. According to the evaluation of the epididymal tissue, the groups treated with A. aculeata pulp exhibited smaller fat deposits compared with the HF diet group. Therefore, we infer that the predominant components in A. aculeata, particularly fibers and MUFAs, promote beneficial effects on health parameters during simultaneous exposure to food rich in saturated fat and cholesterol, typical of the Western diet. This is the first study to correlate the presence of fatty acids from A. aculeata pulp in different proportions added in a HF diet with metabolic and histological parameters in Swiss mice.


Asunto(s)
Dieta Alta en Grasa , Grasas de la Dieta , Adipocitos , Animales , Colesterol , Dieta Alta en Grasa/efectos adversos , Ácidos Grasos Monoinsaturados , Hipertrofia , Lipoproteínas HDL , Ratones
7.
Nutrients ; 12(11)2020 Nov 10.
Artículo en Inglés | MEDLINE | ID: mdl-33182564

RESUMEN

The aim of this study was to evaluate the therapeutic effects of two different doses (250 and 500 mg/kg) of Morinda citrifolia fruit aqueous extract (AE) in high-fat/high-fructose-fed Swiss mice. The food intake, body weight, serum biochemical, oral glucose tolerance test (OGTT), and enzyme-linked immunosorbent assay (ELISA), as well as histological analyses of the liver, pancreatic, and epididymal adipose tissue, were used to determine the biochemical and histological parameters. The chemical profile of the extract was determined by ultra-fast liquid chromatography-diode array detector-tandem mass spectrometry (UFLC-DAD-MS), and quantitative real-time PCR (qRT-PCR) was used to evaluate the gene expressions involved in the lipid and glucose metabolism, such as peroxisome proliferative-activated receptors-γ (PPAR-γ), -α (PPAR-α), fatty acid synthase (FAS), glucose-6-phosphatase (G6P), sterol regulatory binding protein-1c (SREBP-1c), carbohydrate-responsive element-binding protein (ChREBP), and fetuin-A. Seventeen compounds were tentatively identified, including iridoids, noniosides, and the flavonoid rutin. The higher dose of AE (AE 500 mg/kg) was demonstrated to improve the glucose tolerance; however, both doses did not have effects on the other metabolic and histological parameters. AE at 500 mg/kg downregulated the PPAR-γ, SREBP-1c, and fetuin-A mRNA in the liver and upregulated the PPAR-α mRNA in white adipose tissue, suggesting that the hypoglycemic effects could be associated with the expression of genes involved in de novo lipogenesis.


Asunto(s)
Glucosa/metabolismo , Metabolismo de los Lípidos/efectos de los fármacos , Síndrome Metabólico/metabolismo , Morinda/química , Extractos Vegetales/farmacología , Tejido Adiposo , Animales , Dieta Alta en Grasa , Femenino , Fructosa , Regulación de la Expresión Génica/efectos de los fármacos , Glucosa-6-Fosfatasa/metabolismo , Lipogénesis/efectos de los fármacos , Hígado/efectos de los fármacos , Hígado/metabolismo , Masculino , Síndrome Metabólico/inducido químicamente , Síndrome Metabólico/tratamiento farmacológico , Ratones , PPAR alfa/metabolismo , PPAR gamma/metabolismo , Fitoterapia , Extractos Vegetales/uso terapéutico , Proteína 1 de Unión a los Elementos Reguladores de Esteroles/metabolismo
8.
Molecules ; 25(19)2020 Oct 01.
Artículo en Inglés | MEDLINE | ID: mdl-33019745

RESUMEN

Garcinia is a genus of Clusiaceae, distributed throughout tropical Asia, Africa, New Caledonia, Polynesia, and Brazil. Garcinia plants contain a broad range of biologically active metabolites which, in the last few decades, have received considerable attention due to the chemical compositions of their extracts, with compounds which have been shown to have beneficial effects in several diseases. Our work had the objective of reviewing the benefits of five Garcinia species (G. brasiliensis, G. gardneriana, G. pedunculata, G. cambogia, and G. mangstana). These species provide a rich natural source of bioactive compounds with relevant therapeutic properties and anti-inflammatory effects, such as for the treatment of skin disorders, wounds, pain, and infections, having demonstrated antinociceptive, antioxidant, antitumoral, antifungal, anticancer, antihistaminic, antiulcerogenic, antimicrobial, antiviral, vasodilator, hypolipidemic, hepatoprotective, nephroprotective, and cardioprotective properties. This demonstrates the relevance of the genus as a rich source of compounds with valuable therapeutic properties, with potential use in the prevention and treatment of nontransmissible chronic diseases.


Asunto(s)
Garcinia/química , Fitoquímicos/farmacología , Plantas Medicinales/química , Fitoquímicos/química , Especificidad de la Especie
9.
Molecules ; 25(11)2020 Jun 10.
Artículo en Inglés | MEDLINE | ID: mdl-32531999

RESUMEN

There are still controversies regarding the correlation between the beneficial effects for health and the administration of isolated compounds or crude extracts in therapeutic applications. Campomanesia xanthocarpa, found in the Brazilian Cerrado, demonstrated beneficial effects in metabolic disorders associated with obesity. We investigated the effects of Campomanesia xanthocarpa hydroethanolic extract and two isolated substances from the extract (S1 and S2) in a diet-induced obesity (DIO) model. Male Swiss mice were divided into five groups: (1) American Institute of Nutrition (AIN-93M) diet, (2) high-fat diet (HF), (3) HF supplemented with C. xanthocarpa hydroethanolic leaf extract at 100 mg/kg (HFE), (4) HF supplemented with S1 at 1 mg/kg (HFS1) and (5) HF supplemented with S2 at 1 mg/kg (HFS2). The HFS1, HFS2 and HFE groups did not present decreasing body weight or visceral adiposity gain. No differences in glycemic and lipid parameters, or in the expression of protein content in two cytokines, interleukin-6 (IL-6) and anti-inflammatory (IL-10), were observed. Only the HFS1 group displayed decreased food intake. Even though substantial effects such as an improvement in obesity features or the metabolic and histological parameters promoted by S1, S2 and the extract were not observed, further investigations are necessary to evaluate the principal genes and protein expressions involved in regulating food behavior promoted by S1.


Asunto(s)
Dieta Alta en Grasa/efectos adversos , Suplementos Dietéticos , Inflamación/prevención & control , Enfermedades Metabólicas/prevención & control , Myrtaceae/química , Obesidad/prevención & control , Extractos Vegetales/farmacología , Animales , Antiinflamatorios/farmacología , Etanol/química , Inflamación/etiología , Inflamación/metabolismo , Inflamación/patología , Masculino , Enfermedades Metabólicas/etiología , Enfermedades Metabólicas/metabolismo , Enfermedades Metabólicas/patología , Ratones , Obesidad/etiología , Obesidad/metabolismo , Obesidad/patología , Estrés Oxidativo
10.
J Anim Physiol Anim Nutr (Berl) ; 104(5): 1575-1582, 2020 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-32279410

RESUMEN

Non-alcoholic fatty liver disease (NAFLD) is a metabolic disorder caused by excess consumption of hypercaloric foods. Guavira (Campomanesia sp.) pulp has broad technological applicability, yet the peel and seeds are considered industrial residue. The objective of this unprecedented study was to evaluate the effects of the flour from guavira's industrial residue (GF) consumption in rats fed with hypercaloric diet (HD). During 65 days, 50 rats were separated into a control group: 1%, 2%, 4% and 8% HD with GF complementation in the diet. The GF chemical composition, phenolic compounds, antioxidant capacity, serum biochemical parameters (glucose, cholesterol, HDL, non-HDL, triglycerides, AST, ALT, and oral glucose tolerance test), fat liver content, and hepatic histomorphology had been characterized. GF is mainly composed of fibres, with phenolic content of 7,391.09 mg AGE/100 g GF and relevant antioxidant capacity (IC50 2.22 and ORAC 155.68 µmol/TE g-1 ). Serum biochemical analysis did not differ statistically (except ALT reduction, p < .05). Fat liver content was smaller on HD2%GF (p < .0001). The control group and 1% GF showed greater diffuse microvesicular steatosis compared to the other groups when using hepatic morphological analysis (p < .05). GF consumption attenuated NAFLD caused by a hypercaloric diet, and this effect may be related to the fibre content and bioactive compounds in GF.


Asunto(s)
Alimentación Animal/análisis , Ingestión de Energía , Myrtaceae/química , Enfermedad del Hígado Graso no Alcohólico/inducido químicamente , Animales , Dieta/veterinaria , Metabolismo Energético , Hígado/metabolismo , Masculino , Enfermedad del Hígado Graso no Alcohólico/prevención & control , Ratas , Ratas Wistar , Aumento de Peso
11.
J Med Food ; 22(12): 1254-1261, 2019 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-31329496

RESUMEN

The prevalence of gastrointestinal disorders, such as constipation, has been increasing. Genetic factors and lifestyle are some of the etiologies of this issue, affecting the health of the population. Natural products have properties that contribute to health maintenance and health promotion, including reduction of the inflammatory process. Hancornia speciosa, popularly known as mangaba, is an abundant and native fruit in the Brazilian Cerrado, commercialized for culinary purposes and used because of its pharmacological properties. The objective of this study was to evaluate if the supplementation of different concentrations of mangaba pulp can improve intestinal motility and bowel health in Wistar rats. Forty male rats were divided into five groups. The experiment lasted 14 days and the groups were tested with water, industrialized laxative jelly made from tamarind as medication, or mangaba at 5, 10, and 15 mL/kg of body weight. Food intake, weight gain, ion balance, intestinal motility, and histopathological analysis of the small intestine, large intestine, and liver were evaluated. Supplementation of mangaba pulp at its highest concentration (15 mL/kg body weight) caused a 15% increase in the distance traveled by the charcoal meal, and a decrease in serum magnesium levels and white cells in both the small and large intestines. The results suggest that mangaba pulp presents laxative, anti-inflammatory properties and that its consumption is beneficial and should be encouraged.


Asunto(s)
Apocynaceae/química , Motilidad Gastrointestinal/efectos de los fármacos , Extractos Vegetales/farmacología , Animales , Antiinflamatorios/farmacología , Antiinflamatorios/uso terapéutico , Peso Corporal , Brasil , Ingestión de Alimentos , Alimentos Fortificados , Frutas/química , Intestinos/efectos de los fármacos , Intestinos/patología , Laxativos , Hígado/efectos de los fármacos , Hígado/patología , Masculino , Modelos Animales , Extractos Vegetales/uso terapéutico , Ratas , Ratas Wistar , Suero/química , Tamarindus , Aumento de Peso
12.
Nutrients ; 10(6)2018 Jun 20.
Artículo en Inglés | MEDLINE | ID: mdl-29925762

RESUMEN

Polydextrose (PDX) ingestion may increase the intestinal absorption of iron. This study evaluated the effects of 7.5% polydextrose supplementation on markers of iron uptake, transport and storage in partially gastrectomized rats. Half of a batch of 40 male Wistar rats (250 g) underwent Billroth II partial gastrectomy with anterior truncal vagotomy (GXT), while the other half underwent sham gastrectomy (SHAM). At 7 postoperative days, the animals were subdivided into four groups (n = 10): Sham Control and GXT Control (no polydextrose); Sham PDX and GXT PDX (with 7.5% PDX). The animals were euthanized after 60 day of PDX treatment. Organ weight, cecal pH, the characterization and quantification of short-chain fatty acids (SCFA), hematological parameters, hepatic iron content and the expression of ferroportin (FPT) in the jejunum, cecum, colon and liver were evaluated. PDX caused changes in the cecum of the supplemented animals, where there was a decrease in pH, increase in cecal wall and marked production of SCFA, especially acetic and propionic acids (p < 0.05). Hepatic iron levels were lower in GXT animals. PDX increased hemoglobin (HGB) values by 29.2% and hematocrit (HCT) by 55.8% in the GXT PDX group compared to the GXT Control group. The GXT PDX group had lower hepatic FPT expression (p < 0.05). PDX led to increased SCFA concentration in the supplemented animals. Considering that SCFAs play a central role in the increasing nutrients uptake, this mechanism may be involved in altering the hematology profile observed in these animals but not enough to reverse iron deficiency anemia in post-gastrectomy rats.


Asunto(s)
Ácidos Grasos Volátiles/metabolismo , Gastrectomía , Glucanos/farmacología , Hierro/metabolismo , Anemia Ferropénica , Animales , Fibras de la Dieta , Glucanos/administración & dosificación , Hematócrito , Absorción Intestinal/efectos de los fármacos , Masculino , Distribución Aleatoria , Ratas , Ratas Wistar
13.
Rev. Soc. Bras. Med. Trop ; 35(6): 591-595, nov.-dez. 2002. tab
Artículo en Portugués | LILACS | ID: lil-340057

RESUMEN

Cryptococcus neoformans é a levedura capsulada causadora de criptococose em humanos e animais. A variedade neoformans, encontrada em diversas fontes ambientais, inclusive habitats de aves, é importante causa de mortalidade em indivíduos com AIDS em todo o Mundo. Contudo, ainda näo há estudos sobre a sua ecologia na regiäo Centro Oeste brasileira, onde há registro da ocorrência de casos humanos da micose. Para estudar fontes saprofíticas de C. neoformans, na cidade de Campo Grande, foram coletadas 20 amostras de excretas de aves em distintos ambientes. Suspensäo das amostras em salina estéril foram semeadas em placas com meio ágar níger. Após 5 dias, colônias mucóides marrom-escuro foram subcultivadas para identificaçäo através de provas morfofisiológicas, determinaçäo da variedade e sorotipagem. C. neoformans var. neoformans sorotipo A foi isolado de 10 (50 por cento) das amostras, comprovando a ocorrência saprofítica de C. neoformans na cidade de Campo Grande, relacionada a habitat de aves em cativeiro


Asunto(s)
Animales , Femenino , Masculino , Aves/microbiología , Cryptococcus neoformans/aislamiento & purificación , Heces/microbiología , Vivienda para Animales
14.
Rev Soc Bras Med Trop ; 35(6): 591-5, 2002.
Artículo en Portugués | MEDLINE | ID: mdl-12612740

RESUMEN

Cryptococcus neoformans is an encapsulated yeast agent of human and animal cryptococcosis. The variety neoformans is an important cause of mortality in AIDS-patients throughout the world. It has already been isolated worldwide from different environmental sources, including avian habitats. However, in spite of the occurrence of human cases of cryptococcosis, there are no studies on the ecology of this agent in the Brazilian Central-Western region. To study saprophytic sources of C. neoformans in the city of Campo Grande, 20 samples of avian droppings from distinct environments within the city were collected. The samples were suspended in sterile saline and then smeared on niger seed agar medium. Five days later smooth dark-brown colonies were subcultivated for identification by morphophysiologic tests. The variety and serotype was determined. C. neoformans var. neoformans serotype A was isolated from 10 (50%) of the samples collected. Consequently, the saprophytic presence of C. neoformans is related to avian habitats.


Asunto(s)
Aves/microbiología , Cryptococcus neoformans/aislamiento & purificación , Heces/microbiología , Vivienda para Animales , Animales , Femenino , Masculino
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