Antifungal properties of garlic -
The cytokines participation is essential once it increases the fungicide capacity of phagocytes, furthering the fungus destruction Coelho-Castelo et al. The Sporothrix schenckii , causal agent of sporotrichosis is dimorphic fungus, of saprophytic life in soil or plants occasionally infecting people and animals.
In man, most of infections are caused by splinters, wood or wire injuries Marques et al. and FABRIS, V. Esporotricose do gato doméstico Felis catus : transmissão humana. Revista do Instituto de Medicina Tropical de Sao Paulo, vol.
The mycelial form saprophytic of fungus found in the environment grows at temperatures ranging from 25 °C to 30 °C. The yeast form is found in vivo , at 37 °C, where it reproduces by budding Alegranci, ALEGRANCI, P.
The sporotrichosis injuries are usually limited to the skin, subcutaneous tissue, and nearby lymph vessels. Rarely it may spread to other organs or even be systemic, resulting from inhalation of spores. The infection can be classified as skin, cutaneous lymphatic, skin spread, mucosal and systemic Barros et al.
and SCHUBACH, A. Esporotricose: a evolução e os desafios de uma epidemia. Revista Panamericana de Salud Pública, vol. It is the most common subcutaneous mycosis in Latin America, and it has been seen in many countries, in animals and people, it is worth to say that in the 90's in the Rio de Janeiro State the disease reached epidemic proportions Barros et al.
The aim of this study was to evaluate the influence of garlic Allium sativum L. consumption in immunomodulation of healthy and infected Swiss mice induced by S.
Aquous and oily extracts were prepared using 32 g of garlic cloves Allium sativum L. peeled and crushed in mL of distilled water or soybean oil, allowed it to rest for half an hour, and then it was cooled and filtered. The dose of the extracts 0. Peritoneal exudate's cells PECs of Swiis mice, male, adult, healthy or infected with fungus S.
schencki i were collected after the animals were previously submitted to 3. The animals had abdominal skin removed aseptically in a laminar flow cabinet and the peritoneum was exposed. It was inoculated 5 mL of cold PBS with pH 7. The resulting peritoneal fluid was collected and transferred into a conical sterile tube and centrifuged at rpm for 5 minutes.
The cell pellet was washed three times with 3mL of PBS. The cell suspension was adjusted to final concentration of 5x10 6 cells to accomplish the proposed tests, and the plates with PECs were incubated for one hour in CO 2 incubator at 37°C for the formation of the adherent cells layer pad, and after that the supernatant was discarded and the extracts and solutions corresponding the tests were added on cells.
The NO was spectrophotometrically measured by the Griess Reaction Green et al. Analysis of nitrate, nitrite, and [15N]nitrate in biological fluids. Analytical Biochemistry, vol. The determination of the nitric oxide NO releasing was performed on floating PECs from infected and uninfected mice with S.
schenckii fungus by the garlic extracts challenge 1 mg mL -1 and LPS control positive control or RPMIC negative control. The control group did not receive garlic extract by gavage, however these groups of PECs were challenged with two different extracts aqueous and oily.
The IL-1β, IL and IL cytokines in floating were quantified PECs obtained as challenged as described above. Quantitation was done by the ELISA enzyme immunoassay of capturing for each cytokine.
It was used the strain of Sporothrix schenckii isolated from a pulmonary human infection of sporotrichosis Baltimore, MD , courtesy of Reference Materials Laboratory of the Oswaldo Cruz Foundation.
Currently, this isolate is kept in the Clinical Immunology Laboratory of the Department of Clinical Analysis of Pharmaceutical Sciences School of Araraquara - UNESP, in the mycelial stage in Sabouraud broth, at room temperature ± 25 °C. The yeasty form is obtained in BHI Brain Heart Infusion, Difco liquid at 37 °C under constant stirring of rpm during seven days.
Thereafter, an aliquot containing fungal units was transferred to a mL Erlenmeyer containing mL of BHI broth, and cultured for five days under the same conditions, in that it corresponds with the phase of logarithmic growth previously described by Ferreira et al. and CARLOS, I. Optimal clearance of Sporothrix schenckii requires an intact Th17 response in a mouse model of systemic infection.
Immunobiology, vol. The animals were inoculated with µL of a suspension containing 10 6 S. schenckii yeast in PBS or an equal volume of PBS alone by intraperitoneal administration. The evaluation of the systemic fungal load for infections monitoring was made considering a determination of colony forming units CFU in the spleen of the animals after they were removed.
The CFU were counted after 3 days, and affirmed after 5 days of incubation at room temperature. Evaluation of antifungal activity was performed using the microdilution test according to Standard MA2 of National Committee for Clinical Laboratory Standards.
The culture of S. schenckii was used by preparing an inoculum in sterile PBS. Serial dilutions were done in RPMI medium with MOPS, with the fungal suspension adjusted to a concentration of 2. The results were expressed as mean ± standard deviation and were submitted to Lilliefors test to verify the normality, the Levene test to verify the homoscedasticity, and variance analysis ANOVA one way - Tukey post test.
However, animals not previously exposed to extracts CG , when stimulated by the extracts, show more NO production, compared to the negative control, what does not occurs in animals that receive the garlic extract orally AG. This shows that the garlic immunomodulatory activity may be used in different ways, in that the response changes if it is ingested regularly or in acute way Figure 1A.
The AG infected with the S. This shows that the consumption of garlic may increase the production of NO, even in conditions that the production is already increased, as in this case by the induced infection Figure 1B.
The infection increased the NO production in all groups, this is shown in Figures 1C , 1D and 1E. The IL-1β production in infected animals did not differed between the studied groups faced by the challenges with extracts, there was only a difference between the positive control, in that all were different among them, with higher production for the OG.
Clearly, the garlic extracts have showed anti-inflammatory activity when administered in a chronic way 15 days , however, PECs of the animals that did not have previous contact with the extracts CG , when challenged with the garlic extracts, it showed higher values to LPS challenge positive control , indicating that the acute use of garlic may be a potent stimulator of IL-1β production, especially in healthy ones.
Among the healthy animals, those that were given garlic extract had IL production increased Figure 3A , compared to the group that did not received the extract orally CG.
In AG and OG treatments, the LPS challenge, NC and extract corresponding were significantly different. The production of IL by healthy animals that received garlic extract orally AG and OG and were stimulated by the extracts was lower compared to those that did not received CG , however in infected animals the response was unlike, animals from AG and OG stimulated by their extracts show a higher production when compared to the animals of the control group in that it was equally stimulated Figures 4A and 4B.
PECs of CG when challenged with the extracts showed values of IL production above the positive control Figure 4A. The groups treated with garlic showed a higher ability to control the infection, as can be seen in Figure 5. The Table 1 shows the antifungal activity of the extracts at its minimum concentration.
It is known that in natura garlic or in the form of extracts reduces fungal growth, cause damage to the membrane and inhibit the synthesis of lipids, proteins and nucleic acids Harris et al.
and LLOYD, D. Antimicrobial properties of Allium sativum garlic. Applied Microbiology and Biotechnology, vol. and GORJI, A. Garlic: a review of potential therapeutic effects.
Avicenna Journal of Phytomedicine, vol. In this research, the 10 th day post-infection, active washout of fungus, infected animals that were treated with garlic extracts showed much lower fungal load compared to untreated animals, that is, it caused a more effective response in infection control demonstrating effectiveness against in vivo fungus.
In addition, the aqueous and oily extracts showed to be capable of inhibiting S. schenckii growth, respectively, at doses of 0. The most ability in infection control may be justified by the fact that the destruction of invaders by macrophages is significantly increased with the garlic extract elements, which it increases the number of peritoneal macrophages, their action, and expression of receptors in the membranes Ghazanfari et al.
Enhancement of peritoneal macrophage phagocytic activity against by garlic. Leishmania majorAllium Sativum treatmentJournal of Ethnopharmacology, vol. The treatment with garlic then may be indicated as a complementary and alternative way of therapy, thereby decreasing the dose of medication used.
The Immunity against fungus shows up the importance of Th1 pathway, where there is a release of cytokines such as IFN-γ, IL-1 and IL On the other hand, the Th2 pathway that releases stimulating citokines of antibody production IL-4, IL and IL shows limited value in the host defense against these agents Giraldo and Witkin, GIRALDO, P.
and WITKIN, S. Vaginal candidiasis: an incomprehensible challege. Jornal Brasileiro de Doenças Sexualmente Transmissíveis, vol. In the immune response against sporotrichosis, macrophages when activated in culture with fungal cell wall components produce proinflammatory cytokines such as IL-1, which stimulate phagocytic response Carlos et al.
and MAIA, D. Current research on the immune response to experimental sporotrichosis. Mycopathologia, vol. and release intermediate nitrogen and oxygen compounds with fungicide function Carlos et al. and PLACERES, M. Sporothrix schenckii lipid inhibits macrophage phagocytosis: involvement of nitric oxide and tumour necrosis factor-alphaScandinavian Journal of Immunology, vol.
The ability of active macrophages release NO is very important in host defense, as it allows the attack to different types of pathogens Janeway et al.
The same can be seen in this work where animals consuming garlic have showed higher levels of pro-inflammatory interleukins IL-1β and IL when infected, however, when in a healthy state, the anti-inflammatory cytokine IL suffered an increase in the production. Considering that excessive inflammation is a critical factor in many human diseases including cancer, obesity, cardiovascular diseases, neurodegenerative and intestinal diseases, type I diabetes, and even in aging Santangelo et al.
and MASELLA, R. Polyphenols, intracellular signalling and inflammation. Annali dell'Istituto Superiore di Sanità. Also, according to the results of this study, garlic may be used to fight pain, since its consumption decreases the production of IL-1β in healthy animals.
The IL-1β leads to systemic inflammation through the activation of cyclooxygenase-2, with PGE2 formation in the anterior hypothalamus, having an important role in the development of pain Oliveira et al. Garlic samples were purchased from Dutsinma central market, Katsina state.
The samples were washed, separated and peeled to obtain the edible portion. The fungi were isolated using the culture method and identified based on morphological characteristics. The extract was prepared using two solvents aqueous and ethanol by soaking method. The antifungal activity of aqueous and ethanolic garlic extract was determined on some selected fungi namely, Fusarium spp and Rhizopus spp.
From the results it is clear that, ethanol extract showed more activity when compare to aqueous extract. The diameter of zones of inhibition for the ethanolic extract ranged between 4.
The MIC for the ethanolic extract was 2. Das, K. Reactive oxygen species ROS and response of antioxidants as ROS-scavengers during environmental stress in plants. Das, N. Microbial degradation of petroleum hydrocarbon contaminants: an overview.
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Acta Agric. Liu, N. Antioxidant enzymes regulate reactive oxygen species during pod elongation in pisum sativum and brassica chinensis. PLoS ONE 9:e Liu, R. Histological responses to downy mildew in resistant and susceptible grapevines. Protoplasma , — Liu, Y. Potato dextrose agar antifungal susceptibility testing for yeasts and molds: evaluation of phosphate effect on antifungal activity of CMT Ma, Z.
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Zhi-hui, C. Allelopathic effects of decomposing garlic stalk on some vegetable crops. Keywords : Allicin, genetic diversity, antifungal, biostimulant, antioxidant enzymes, HPLC. Citation: Hayat S, Cheng Z, Ahmad H, Ali M, Chen X and Wang M Garlic, from Remedy to Stimulant: Evaluation of Antifungal Potential Reveals Diversity in Phytoalexin Allicin Content among Garlic Cultivars; Allicin Containing Aqueous Garlic Extracts Trigger Antioxidants in Cucumber.
Received: 04 February ; Accepted: 03 August ; Published: 25 August Copyright © Hayat, Cheng, Ahmad, Ali, Chen and Wang. This is an open-access article distributed under the terms of the Creative Commons Attribution License CC BY.
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ORIGINAL RESEARCH article Front. Garlic, from Remedy to Stimulant: Evaluation of Antifungal Potential Reveals Diversity in Phytoalexin Allicin Content among Garlic Cultivars; Allicin Containing Aqueous Garlic Extracts Trigger Antioxidants in Cucumber. Materials and Methods Garlic Cultivars, Aqueous Extract Preparation and Allicin Quantification Fresh, uniform sized bulbs of 28 garlic cultivars were selected from the garlic germplasm NWSUAF Yangling, Shaanxi, China.
Fungal Strains Four fungal species were employed in this research; Botrytis cinerea, Fusarium oxysporum, Phytophthora capsici , and Verticillium dahliae.
Alireza Khodavandi, Fahimeh Alizadeh, Nabil S. Harmal, Shiran M. Thyroid health catechins efficacy of allicin compared with Antifungwl in barlic systemic Hormonal balance supplements albicans infections was evaluated both in vitro and in vivo through a systemic candidiasis mouse model. Determination of in vitro minimum inhibitory concentrations MICs for different C. albicans isolates revealed that both allicin and fluconazole showed different MICs that ranged from 0.Video
Garlic and the skin Thank Antivungal for visiting nature. You are Insulin pump therapy accuracy a browser Anitfungal with limited support for CSS. To obtain the best experience, we recommend you use a more Thyroid health catechins to Antifugal browser or turn Antfungal compatibility mode Protein and athletic performance goals Thyroid health catechins Explorer. In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript. The antifungal activity, kinetics and molecular mechanism of action of garlic oil against Candida albicans were investigated in this study using multiple methods. Using the poisoned food technique, we determined that the minimum inhibitory concentration of garlic oil was 0. Observation by transmission electron microscopy indicated that garlic oil could penetrate the cellular membrane of C.
Antifungal properties of garlic -
The gr1 mutant line is a knockout mutant of glutathione reductase and has a higher proportion of GSSG in the glutathione pool because it cannot reduce GSSG back to GSH As can be seen in Fig.
This was not observed in the wt and this phenomenon may be worthy of further investigation. The concentration-dependent inhibition of root growth by thiosulfinates is shown in Fig. The enhanced sensitivity of GSH metabolism mutants compared to the Col-0 wt confirms the results of the chemogenetic screen with yeast mutants Figs.
Effect of thiosulfinates on Arabidopsis root growth. MSDS was purchased from Acros Chemicals Geel, Belgium. was purchased from Carl Roth Karlsruhe, Germany.
was purchased from Carl Roth. TLC was performed using Merck TLC Silica gel 60 F with concentration zone. Solvent B ethyl acetate Liquid chromatography was performed using silica gel 60 0. The synthetic procedure for thiosulfinates was after Albrecht et al. Dimethyldisulfide 1.
The organic layer was separated, dried with Na 2 SO 4 and filtered. The solvent was evaporated under reduced pressure at room temperature to yield a clear oily substance of characteristic smell. Yield: 1. DMTS synthesis was stoichiometric, yielding a pure product with a single HPLC peak and no further purification was carried out.
Diethyldisulfide 2. The solvent was evaporated under reduced pressure at room temperature and the residue was dissolved in an ethyl acetate: n-hexane mixture The product was purified on a silica gel 60 column with the same mixture used as mobile phase.
The product-containing fractions were combined, and the solvents were removed by rotary evaporation at room temperature. The product was a clear oil with a pungent smell. Dipropyldisulfide 2. The combined organic phases were washed with saturated Na 2 CO 3 solution, dried over Na 2 SO 4 and filtered.
The solvent was removed under reduced pressure at room temperature to yield a clear oil. Yield: 2. DPTS synthesis was stoichiometric, yielding a pure product with a single HPLC peak and no further purification was carried out. The solvent was removed under reduced pressure and the product was dissolved in the eluent, a mixture of n-hexane and ethyl acetate Fractions were collected into tubes cooled in an ice bath and TLC was used to identify fractions containing only allicin.
Those fractions were combined, dried with anhydrous sulfates e. Na 2 SO 4 , MgSO 4 or CuSO 4 and filtered.
The solvents were removed under reduced pressure at RT to yield a clear, oily substance that smells like garlic. Dibenzyldisulfide 0. The substance was dissolved in a mixture of ethyl acetate and n-hexane and purified on a silica gel column. The product-containing fractions were combined, and the solvents removed under reduced pressure at RT.
The residual white powder was characterized by NMR as dibenzylthiosulfinate. Yield: 0. A Jasco HPLC system with a UV multichannel detector and a PU pump was used.
The mobile phase consisted of H 2 O A and methanol. The chemical decomposition of the compounds was analysed with HPLC using UV detection. coli K12, Pseudomonas fluorescens and Pseudomonas syringae pv. phaseolicola were used as representative Gram-negative bacteria. The haploid Saccharomyces cerevisiae yeast strain BY Matα; his3Δ1;leu2Δ0, lys2Δ0, ura3Δ0 was used as a model fungus in tests with different thiosulfinates.
The BY mutant Δ yap1 YMLw used in this study lacks a redox-sensitive transcription factor that is important for oxidative stress response. Yeast was grown in CSM medium 0. Susceptibility testing was performed following European Committee on Antimicrobial Susceptibility Testing EUCAST guidelines using the broth dilution method in well microtiter plate format Bacteria Escherichia coli , Pseudomonas syringae pv.
phaseolicola , Pseudomonas fluorescens Pf- 01, Micrococcus luteus were grown over-night in media and temperatures as described above.
At the next day, the bacteria were diluted to an OD of 0. The lowest concentrations that showed no visible growth of bacteria is the minimal inhibitory concentration MIC. An overnight culture of the bacteria grown in LB-medium under optimal conditions E. The diameter of the inhibition zone was measured and plates were photographed.
Further procedure was as with bacteria see above. Bacteria were seeded into LB-medium as described above. The Petri dish was incubated upside down over night at the particular temperature E. For a time resolved test the droplet was removed after following hours passed: 0.
It was optically evaluated to which dilution yeast was able to grow. The compounds tested were diluted in CSM medium to final concentrations as indicated in the figure. Louis, USA. Thiosulfinate concentrations between 0. BY-2 cells kindly provided by Dr. Langenbach, Institut f. Biologie III, RWTH Aachen University were grown in modified MS Medium 4.
Data are presented as means with standard deviations of four replicates. Arabidopsis seedling root assay was performed after Borlinghaus et al. The Petri plates were tilted to an angle of approx.
After three days of cultivation, filter papers were transferred to MS medium that contained different amounts of thiosulfinates. After 3 days treatment seedlings were photographed and the root length was measured. Based on allicin diallylthiosulfinate, DATS as the lead compound, a comprehensive investigation of the antibiotic properties of molecules with a thiosulfinate functional group and symmetrical side chains modifying their physical properties log P , diffusivity showed that:.
All thiosulfinates tested exhibit a degree of antimicrobal activity comparable to allicin, but absolute activities depended on the conditions used in the individual tests.
For allicin, activity by diffusion through the gas phase could be detected after as little as one hour of exposure. This may suggest a possible use for the treatment of lung diseases because conventional antibiotics are not volatile.
Furthermore, uses in agriculture, e. fumigation of soils and treatment of seeds against seed-borne diseases can be considered. The hypersusceptibility of the Saccharomyces cerevisiae Δyap1 , Δglr1 , Δzwf1 , Δgnd1 , and Δtrx2 and the Arabidopsis pad2 and gr1 mutants suggests that, like allicin, the other thiosulfinates may attack the glutathione pool and glutathione metabolism in target cells.
The synthesis method employed 32 can be used for very many natural and non-natural disulfides to create the more reactive electrophilic sulfur centre of the thiosulfinate functional group and opens up the possibility to investigate very many novel compounds, with potentially improved properties compared to the lead substance allicin.
Although allicin and the analogue thiosulfinates are natural products, nearly all to be found to greater or lesser extent in the common foodstuff garlic, any strategies to develop thiosulfinate-containing formulations for use in medicine and agriculture must of course be accompanied by detailed economic feasibility analysis and toxicology testing, not least of all from the view of potential exposure of agricultural workers when plants are being treated on a field scale.
The results presented here contribute to knowledge of the effects of thiosulfinates on a wide range of organisms including procaryotes and eucaryotes and suggest that thiosulfinates, alone or possibly in combination with other substances, could be developed for specific applications in medicine or agriculture.
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Download references. Financial support from the RWTH Aachen University MCHG, NH, RL, AJS and the University of the Saarland, Saarbrücken RL, CJ is gratefully acknowledged. Ulrike Noll is gratefully acknowledged for excellent technical support. Department of Plant Physiology, RWTH Aachen University, , Aachen, Germany.
Roman Leontiev, Nils Hohaus, Martin C. Division of Bioorganic Chemistry, School of Pharmacy, Saarland University, , Saarbrücken, Germany. You can also search for this author in PubMed Google Scholar. performed the experiments, prepared the Figures and wrote parts of the manuscript, N.
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Skip to main content Thank you for visiting nature. nature scientific reports articles article. Download PDF. Subjects Antibiotics Small molecules. Abstract Allicin diallylthiosulfinate is a defence molecule from garlic Allium sativum L.
Introduction Garlic has been used since ancient times for its health beneficial properties and modern research has provided a scientific basis for this practice 1 , 2 , 3. Figure 1. Full size image. Figure 2.
Results and Discussion Comparison of the physical properties of DMTS, DETS, DATS, DPTS and DBTS The structures and the physical properties of the test substances dimethyl- DMTS , diethyl- DETS , diallyl- DATS, allicin , dipropyl- DPTS and dibenzyl- DBTS thiosulfinate are summarized in Fig.
Its chemical structure is very complex and perhaps is made as a self-protection way against micro-organisms and other threats mainly present in soil Amagase et al. and ITAKURA, Y. Intake of garlic and its bioactive components. The Journal of Nutrition, vol.
Among its ingredients with biological activity, perhaps the most important are the organosulfur compounds that are present in mg per gram of fresh garlic. Garlic contains three times more sulfur than onion, in addition to arginine, oligosaccharides, flavonoids and selenium Holub et al.
and DONATO, F. Bioactive compounds and cancer. Its taste and burning characteristics are because of the hydrolysis of S-alk compounds en yl cysteine sulphoxides ACSOs by alliinase enzyme Randle and Lancaster, RANDLE, W. and LANCASTER, J. Sulphur compounds in alliums in relation to flavour quality.
In: H. CURRAH, eds. Allium crop science: recent advances. CAB International, pp. The alliin S-alilcisteína S-oxide is the main organsulfur ingredient identified in intact garlic bulbs, as when they are cut or grinded, the alliin is changed to allicin by alliinase.
The chemical changing of allicin by the heat it yields other sulfur compounds such as ajoenos, vinilditiina and methyl allyl trisulfide Ankri and Mirelman, ANKRI, S.
and MIRELMAN, D. Antimicrobial properties of allicin from garlic. Microbes and Infection, vol. Nutrition plays an important role in modulating the immune and inflammatory response, as the nutrients regulate cellular and humoral defensing systems, by the changing in the formation of mediators or by interfering with the transduction pathways of cellular signals, altering the balance between cytokines, and lessen depletion of tissue nutrients Parham, PARHAM, P.
O sistema imune. Porto Alegre: ArtMed. Several cytokines produced by leukocytes are called interleukins. Interleukin-1 IL-1 , for instance, mediates inflammation and is produced mainly by activated macrophages. Its effects include stimulating the production of chemokines, increasing production of neutrophils by bone marrow, and increasing inflammatory activity.
However, this process can be highly regulated by antagonist action such as interleukin IL Correa-Camacho et al. and SOARES, A.
Atherosclerosis, an inflammatory response. Arq Ciênc Saúde, vol. and BERTRAM, K. Porto Alegre: AMGH. The IL suppresses the activity of activated macrophages, T cells and Natural killer cells NK , and also causes inhibition of interleukin 12 IL in macrophages, which is the main form of NK cells activation.
The IL stimulates production of IFN-y by NK cells and T lymphocytes, enhances cytotoxicity mediated by NK cells and stimulates differentiation of T cells.
Although many other cells appear to secrete IL, only macrophages and dendritic cells yield the biologically active cytokine Abbas et al. and PILLAI, S. Imunologia celular e molecular. Rio de Janeiro: Elsevier Brasil. and SALOMÃO, R. Citocinas e dor. Revista Brasileira de Anestesiologia, vol.
Macrophages play a basic key role in defending the body system, by phagocytosis, bringing forth free radicals, mediation of inflammation process and secretion of diverse substances Fischer et al.
and VIDOR, T. Imunomodulação pela própolis. Arquivos do Instituto Biologico, vol. It belongs to the mononuclear phagocyte system, and are a part, along with NK cells, of the first line of defense against pathogens Klimp et al.
and DAEMEN, T. A potential role of macrophage activation in the treatment of cancer. They are classified as M1 and M2, being M1 producer of the nitric oxide synthase enzyme, so it puts down L-arginine, yelding nitric oxide NO , vasodilator that also modulates inflammatory or anti-inflammatory reactions, depending on cell type and stimulus Alegranci, ALEGRANCI, P.
Análise dos macrófagos M1 e M2 durante a infecção por Sporothrix schenckii em modelo murino. Araraquara: Universidade Estadual Paulista, 97 p. PhD Thesis. and YOSHIDA, W. Óxido nítrico: revisão. Acta Cirurgica Brasileira, vol. and KYO, E. Aged garlic extract enhances production of nitric oxide.
Life Sciences, vol. Fungal infections are a significant cause of morbidity and mortality in humans. They are considered opportunistic as the etiologic agents cause mild illness or no illness in healthy people, but it may infect and cause serious diseases in immunosuppressed ones.
A simple infection such as candidiasis may be severe and invasive, colonizing the esophagus, stomach and intestine in patients with immunodeficiency syndrome Abbas et al.
and LORENZI, J. Immune response to infectious diseases. Medicina, vol. and DONALD CAPRA, J. Porto Alegre: Artes Médicas Sul. Phagocytes, mainly macrophages, respond against fungi and mycoses through the enzyme NO-induced and other mediators secreted to destroy these pathogens.
The cytokines participation is essential once it increases the fungicide capacity of phagocytes, furthering the fungus destruction Coelho-Castelo et al. The Sporothrix schenckii , causal agent of sporotrichosis is dimorphic fungus, of saprophytic life in soil or plants occasionally infecting people and animals.
In man, most of infections are caused by splinters, wood or wire injuries Marques et al. and FABRIS, V. Esporotricose do gato doméstico Felis catus : transmissão humana. Revista do Instituto de Medicina Tropical de Sao Paulo, vol. The mycelial form saprophytic of fungus found in the environment grows at temperatures ranging from 25 °C to 30 °C.
The yeast form is found in vivo , at 37 °C, where it reproduces by budding Alegranci, ALEGRANCI, P. The sporotrichosis injuries are usually limited to the skin, subcutaneous tissue, and nearby lymph vessels.
Rarely it may spread to other organs or even be systemic, resulting from inhalation of spores. The infection can be classified as skin, cutaneous lymphatic, skin spread, mucosal and systemic Barros et al.
and SCHUBACH, A. Esporotricose: a evolução e os desafios de uma epidemia. Revista Panamericana de Salud Pública, vol. It is the most common subcutaneous mycosis in Latin America, and it has been seen in many countries, in animals and people, it is worth to say that in the 90's in the Rio de Janeiro State the disease reached epidemic proportions Barros et al.
The aim of this study was to evaluate the influence of garlic Allium sativum L. consumption in immunomodulation of healthy and infected Swiss mice induced by S.
Aquous and oily extracts were prepared using 32 g of garlic cloves Allium sativum L. peeled and crushed in mL of distilled water or soybean oil, allowed it to rest for half an hour, and then it was cooled and filtered. The dose of the extracts 0. Peritoneal exudate's cells PECs of Swiis mice, male, adult, healthy or infected with fungus S.
schencki i were collected after the animals were previously submitted to 3. The animals had abdominal skin removed aseptically in a laminar flow cabinet and the peritoneum was exposed.
It was inoculated 5 mL of cold PBS with pH 7. The resulting peritoneal fluid was collected and transferred into a conical sterile tube and centrifuged at rpm for 5 minutes. The cell pellet was washed three times with 3mL of PBS.
The cell suspension was adjusted to final concentration of 5x10 6 cells to accomplish the proposed tests, and the plates with PECs were incubated for one hour in CO 2 incubator at 37°C for the formation of the adherent cells layer pad, and after that the supernatant was discarded and the extracts and solutions corresponding the tests were added on cells.
The NO was spectrophotometrically measured by the Griess Reaction Green et al. Analysis of nitrate, nitrite, and [15N]nitrate in biological fluids.
Analytical Biochemistry, vol. The determination of the nitric oxide NO releasing was performed on floating PECs from infected and uninfected mice with S. schenckii fungus by the garlic extracts challenge 1 mg mL -1 and LPS control positive control or RPMIC negative control.
The control group did not receive garlic extract by gavage, however these groups of PECs were challenged with two different extracts aqueous and oily. The IL-1β, IL and IL cytokines in floating were quantified PECs obtained as challenged as described above.
Quantitation was done by the ELISA enzyme immunoassay of capturing for each cytokine. It was used the strain of Sporothrix schenckii isolated from a pulmonary human infection of sporotrichosis Baltimore, MD , courtesy of Reference Materials Laboratory of the Oswaldo Cruz Foundation.
Currently, this isolate is kept in the Clinical Immunology Laboratory of the Department of Clinical Analysis of Pharmaceutical Sciences School of Araraquara - UNESP, in the mycelial stage in Sabouraud broth, at room temperature ± 25 °C.
The yeasty form is obtained in BHI Brain Heart Infusion, Difco liquid at 37 °C under constant stirring of rpm during seven days. Thereafter, an aliquot containing fungal units was transferred to a mL Erlenmeyer containing mL of BHI broth, and cultured for five days under the same conditions, in that it corresponds with the phase of logarithmic growth previously described by Ferreira et al.
and CARLOS, I. Optimal clearance of Sporothrix schenckii requires an intact Th17 response in a mouse model of systemic infection. Immunobiology, vol. The animals were inoculated with µL of a suspension containing 10 6 S. schenckii yeast in PBS or an equal volume of PBS alone by intraperitoneal administration.
The evaluation of the systemic fungal load for infections monitoring was made considering a determination of colony forming units CFU in the spleen of the animals after they were removed.
The CFU were counted after 3 days, and affirmed after 5 days of incubation at room temperature. Evaluation of antifungal activity was performed using the microdilution test according to Standard MA2 of National Committee for Clinical Laboratory Standards.
The culture of S. schenckii was used by preparing an inoculum in sterile PBS. Serial dilutions were done in RPMI medium with MOPS, with the fungal suspension adjusted to a concentration of 2.
The results were expressed as mean ± standard deviation and were submitted to Lilliefors test to verify the normality, the Levene test to verify the homoscedasticity, and variance analysis ANOVA one way - Tukey post test.
However, animals not previously exposed to extracts CG , when stimulated by the extracts, show more NO production, compared to the negative control, what does not occurs in animals that receive the garlic extract orally AG. This shows that the garlic immunomodulatory activity may be used in different ways, in that the response changes if it is ingested regularly or in acute way Figure 1A.
The AG infected with the S. This shows that the consumption of garlic may increase the production of NO, even in conditions that the production is already increased, as in this case by the induced infection Figure 1B. The infection increased the NO production in all groups, this is shown in Figures 1C , 1D and 1E.
The IL-1β production in infected animals did not differed between the studied groups faced by the challenges with extracts, there was only a difference between the positive control, in that all were different among them, with higher production for the OG.
Clearly, the garlic extracts have showed anti-inflammatory activity when administered in a chronic way 15 days , however, PECs of the animals that did not have previous contact with the extracts CG , when challenged with the garlic extracts, it showed higher values to LPS challenge positive control , indicating that the acute use of garlic may be a potent stimulator of IL-1β production, especially in healthy ones.
Among the healthy animals, those that were given garlic extract had IL production increased Figure 3A , compared to the group that did not received the extract orally CG. In AG and OG treatments, the LPS challenge, NC and extract corresponding were significantly different.
The production of IL by healthy animals that received garlic extract orally AG and OG and were stimulated by the extracts was lower compared to those that did not received CG , however in infected animals the response was unlike, animals from AG and OG stimulated by their extracts show a higher production when compared to the animals of the control group in that it was equally stimulated Figures 4A and 4B.
PECs of CG when challenged with the extracts showed values of IL production above the positive control Figure 4A. The groups treated with garlic showed a higher ability to control the infection, as can be seen in Figure 5.
The Table 1 shows the antifungal activity of the extracts at its minimum concentration. It is known that in natura garlic or in the form of extracts reduces fungal growth, cause damage to the membrane and inhibit the synthesis of lipids, proteins and nucleic acids Harris et al.
and LLOYD, D. Antimicrobial properties of Allium sativum garlic. Applied Microbiology and Biotechnology, vol. and GORJI, A. Garlic: a review of potential therapeutic effects. Avicenna Journal of Phytomedicine, vol. In this research, the 10 th day post-infection, active washout of fungus, infected animals that were treated with garlic extracts showed much lower fungal load compared to untreated animals, that is, it caused a more effective response in infection control demonstrating effectiveness against in vivo fungus.
In addition, the aqueous and oily extracts showed to be capable of inhibiting S. schenckii growth, respectively, at doses of 0. The most ability in infection control may be justified by the fact that the destruction of invaders by macrophages is significantly increased with the garlic extract elements, which it increases the number of peritoneal macrophages, their action, and expression of receptors in the membranes Ghazanfari et al.
Enhancement of peritoneal macrophage phagocytic activity against by garlic. Leishmania majorAllium Sativum treatmentJournal of Ethnopharmacology, vol.
The treatment with garlic then may be indicated as a complementary and alternative way of therapy, thereby decreasing the dose of medication used.
The Immunity against fungus shows up the importance of Th1 pathway, where there is a release of cytokines such as IFN-γ, IL-1 and IL On the other hand, the Th2 pathway that releases stimulating citokines of antibody production IL-4, IL and IL shows limited value in the host defense against these agents Giraldo and Witkin, GIRALDO, P.
Insulin pump therapy accuracy study was aimed to determine the antifungal Antifungal properties of garlic of garlic on Antifunggal selected Organic Refreshment Choices. Garlic Thyroid health catechins were purchased from Antifungla central ov, Katsina propertiew. The galic were washed, separated and peeled to obtain the edible portion. The fungi were isolated using the culture method and identified based on morphological characteristics. The extract was prepared using two solvents aqueous and ethanol by soaking method. The antifungal activity of aqueous and ethanolic garlic extract was determined on some selected fungi namely, Fusarium spp and Rhizopus spp. From the results it is clear that, ethanol extract showed more activity when compare to aqueous extract.
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