RESUMO
The present study evaluated the productive characteristics, nutrient digestibility, and economic indexes of broiler chickens submitted to diets containing different nutritional levels, and identified advantages for the commercialization of poultry in the griller-type whole chicken model. A total of 180 COBB 500 chicks ™ were distributed in a completely randomized design, in a 2x3 factorial scheme: sex (male and female) and nutritional level (nutritional requirements for male, female or mixed flock); with 6 repetitions of 5 birds each. It was observed effect of the sex factor in the performance and the economic analysis, in which the male chickens presented the highest feed costs, in spite of exhibiting the best feed conversion rates and gross marketability, resulting from greater body weight. Considering nutritional levels, the most outstanding diet was that formulated according to the demands of the females, which did not affect the performance or carcass characteristics of the birds, obtained the highest gross margins with good feeding costs, showed good retention of dry matter and gross energy, regardless of sex. The diet of nutritional requirements of the females provides promising results for the production of broiler chickens. The female birds present marketing advantages in the griller-type chicken model.
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Fenômenos Fisiológicos da Nutrição Animal , Galinhas , Animais , Feminino , Masculino , Ração Animal/análise , Peso Corporal , Dieta/veterinária , Suplementos Nutricionais/análise , Digestão , NutrientesRESUMO
Invasive fungal infections (IFIs) caused by Candida species are an emerging threat globally, given that patients at-risk and antifungal resistance are increasing. Antimicrobial peptides (AMPs) have shown good therapeutic capacity against different multidrug-resistant (MDR) microorganisms. This study evaluated the activity of the synthetic peptide, PNR20, against Candida albicans ATCC 10231 and a MDR Colombian clinical isolate of Candida auris. Perturbation of yeast cell surface was evaluated using scanning electron microscopy. Cell viability of Vero cells was determined to assess peptide toxicity. Additionally, survival, fungal burden, and histopathology of BALB/c mice infected intravenously with each Candida species and treated with PNR20 were analyzed. Morphological alterations were identified in both species, demonstrating the antifungal effect of PNR20. In vitro, Vero cells' viability was not affected by PNR20. All mice infected with either C. albicans or C. auris and treated with PNR20 survived and had a significant reduction in the fungal burden in the kidney compared to the control group. The histopathological analysis in mice infected and treated with PNR20 showed more preserved tissues, without the presence of yeast, compared to the control groups. This work shows that the utilization of PNR20 is a promising therapeutic alternative against disseminated candidiasis.
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Candidiasis is an opportunistic infection affecting immunosuppressed and hospitalized patients, with mortality rates approaching 40% in Colombia. The growing pharmacological resistance of Candida species and the emergence of multidrug-resistant Candida auris are major public health problems. Therefore, different antimicrobial peptides (AMPs) are being investigated as therapeutic alternatives to control candidiasis effectively and safely. This work aimed to evaluate the in vitro antifungal activity of three synthetic AMPs, PNR20, PNR20-1, and 35409, against ATCC reference strains of Candida albicans, Candida glabrata, Candida parapsilosis, Candida krusei, and Candida tropicalis, and clinical isolates of C. auris. Antifungal susceptibility testing, determined by broth microdilution, showed that the AMPs have antifungal activity against planktonic cells of all Candida species evaluated. In C. auris and C. albicans, the peptides had an effect on biofilm formation and cell viability, as determined by the XTT assay and flow cytometry, respectively. Also, morphological alterations in the membrane and at the intracellular level of these species were induced by the peptides, as observed by transmission electron microscopy. In vitro, the AMPs had no cytotoxicity against L929 murine fibroblasts. Our results showed that the evaluated AMPs are potential therapeutic alternatives against the most important Candida species in Colombia and the world.
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Salmonella enterica is a pathogen capable of colonizing various environments, including the intestinal tract of different animals such as mammals, birds, and reptiles, which can act as carriers. S. enterica infection induces different clinical diseases, gastroenteritis being the most common, which in some cases, can evolve to septicemia and meningitis. Reptiles and amphibians have been reported as a reservoir of Salmonella, and transmission of the pathogen to humans has been documented. This study aimed to determine the presence of virulence genes and characterize the genotypic antibiotic resistance profile in Salmonella strains isolated from Caiman crocodilus fuscus obtained in situ (natural habitat) in Prado, Tolima, Colombia in a previous study and stored in a strain bank in our laboratory. Fifteen Salmonella strains were evaluated through endpoint PCR to determine the presence of resistance genes and virulence genes. The genes blaTEM, strB, and sul1 were detected in all the strains that confer resistance to ampicillin, streptomycin, and sulfamethoxazole, as well as the virulence genes invA, pefA, prgH, spaN, tolC, sipB, sitC, pagC, msgA, spiA, sopB, sifA, lpfA, csgA, hilA, orgA, iroN, avrA, and sivH, indicating the possible role of babilla (Caiman crocodilus fuscus) as a carrier of multidrug-resistant bacteria.
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Fungal infections have increased in recent decades with considerable morbidity and mortality, mainly in immunosuppressed or admitted-to-the-ICU patients. The fungal resistance to conventional antifungal treatments has become a public health problem, especially with Candida that presents resistance to several antifungals. Therefore, generating new alternatives of antifungal therapy is fundamental. One of these possibilities is the use of antimicrobial peptides, such as LL-37, which acts on the disruption of the microorganism membrane and promotes immunomodulatory effects in the host. In this study, we evaluated the in vitro antifungal activity of the LL-37 analogue peptides (AC-1, LL37-1, AC-2, and D) against different Candida spp. and clinical isolates obtained from patients with vulvovaginal candidiasis. Our results suggest that the peptides with the best ranges of MICs were LL37-1 and AC-2 (0.07 µM) against the strains studied. This inhibitory effect was confirmed by analyzing the yeast growth curves that evidenced a significant decrease in the fungal growth after exposure to LL-37 peptides. By the XTT technique we observed a significant reduction in the biofilm formation process when compared to yeasts untreated with the analogue peptides. In conclusion, we suggest that LL-37 analogue peptides may play an important antimicrobial role against Candida spp.
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The repurposing strategy was applied herein to evaluate the effects of lopinavir, an aspartic protease inhibitor currently used in the treatment of HIV-infected individuals, on the globally widespread opportunistic human fungal pathogen Candida albicans by using in silico, in vitro and in vivo approaches in order to decipher its targets on fungal cells and its antifungal mechanisms of action. Secreted aspartic proteases (Saps) are the obviously main target of lopinavir. To confirm this hypothesis, molecular docking assays revealed that lopinavir bound to the Sap2 catalytic site of C. albicans as well as inhibited the Sap hydrolytic activity in a typically dose-dependent manner. The inhibition of Saps culminated in the inability of C. albicans yeasts to assimilate the unique nitrogen source (albumin) available in the culture medium, culminating with fungal growth inhibition (IC50 = 39.8 µM). The antifungal action of lopinavir was corroborated by distinct microscopy analyses, which evidenced drastic and irreversible changes in the morphology that justified the fungal death. Furthermore, our results revealed that lopinavir was able to (i) arrest the yeasts-into-hyphae transformation, (ii) disturb the synthesis of neutral lipids, including ergosterol, (iii) modulate the surface-located molecules, such as Saps and mannose-, sialic acid- and N-acetylglucosamine-containing glycoconjugates, (iv) diminish the secretion of hydrolytic enzymes, such as Saps and esterase, (v) negatively influence the biofilm formation on polystyrene surface, (vi) block the in vitro adhesion to epithelial cells, (vii) contain the in vivo infection in both immunocompetent and immunosuppressed mice and (viii) reduce the Sap production by yeasts recovered from kidneys of infected animals. Conclusively, the exposed results highlight that lopinavir may be used as a promising repurposing drug against C. albicans infection as well as may be used as a lead compound for the development of novel antifungal drugs.
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BACKGROUND: Candida glabrata yeast is the second cause of candidiasis worldwide. Differs from other yeasts since assimilates only glucose and trehalose (a characteristic used in rapid identification tests for this pathogen) by secreting into the medium a highly active acid trehalase encoded by the CgATH1 gene. OBJECTIVE: This study aimed to characterise the function of the acid trehalase in the physiopathology of C. glabrata. METHODS: Gene deletion was performed to obtain a mutant ath1Δ strain, and the ability of the ath1Δ strain to grow in trehalase, or the presence of trehalase activity in the ath1Δ yeast cells, was verified. We also tested the virulence of the ath1Δ strain in a murine model of infection. FINDINGS: The ath1Δ mutant strain grows normally in the presence of glucose, but loses its ability to grow in trehalose. Due to the high acid trehalase activity present in wild-type cells, the cytoplasmic neutral trehalase activity is only detected in the ath1Δ strain. We also observed a significantly lower virulence of the ath1Δ strain in a murine model of infection with either normal or immunocompromised mice. MAIN CONCLUSIONS: The acid trehalase is involved in the hydrolysis of external trehalose by C. glabrata, and the enzyme also plays a major virulence role during infectivity.
Assuntos
Candida glabrata/genética , Trealase/metabolismo , Virulência/genética , Animais , Candida glabrata/metabolismo , Candida glabrata/patogenicidade , Candida glabrata/fisiologia , Candidíase , Deleção de Genes , Genes Fúngicos , Hidrolases , Camundongos , Trealase/genética , Trealase/fisiologia , Trealose/análise , Virulência/fisiologiaRESUMO
Commensal yeast from the genus Candida is part of the healthy human microbiota. In some cases, Candida spp. dysbiosis can result in candidiasis, the symptoms of which may vary from mild localized rashes to severe disseminated infections. The most prevalent treatments against candidiasis involve fluconazole, itraconazole, miconazole, and caspofungin. Moreover, amphotericin B associated with prolonged azole administration is utilized to control severe cases. Currently, numerous guidelines recommend echinocandins to treat invasive candidiasis. However, resistance to these antifungal drugs has increased dramatically over recent years. Considering this situation, new therapeutic alternatives should be studied to control candidiasis, which has become a major medical concern. Limonene belongs to the group of terpene molecules, known for their pharmacological properties. In this study, we evaluated in vitro the limonene concentration capable of inhibiting the growth of yeast from the genus Candida susceptible or resistant to antifungal drugs and its capacity to induce fungal damage. In addition, intravaginal fungal infection assays using a murine model infected by Candida albicans were carried out and the fungal burden, histopathology, and scanning electron microscopy were evaluated. All of our results suggest that limonene may play a protective role against the infection process by yeast from the genus Candida.
RESUMO
Candida auris and Candida haemulonii complex (C. haemulonii, C. haemulonii var. vulnera and C. duobushaemulonii) are phylogenetically related species that share some physiological features and habits. In the present study, we compared the virulence of these yeast species using two different experimental models: (i) Galleria mellonella larvae to evaluate the survival rate, fungal burden, histopathology and phagocytosis index and (ii) BALB/c mice to evaluate the survival. In addition, the fungal capacity to form biofilm over an inert surface was analyzed. Our results showed that in both experimental models, the animal survival rate was lower when infected with C. auris strains than the C. haemulonii species complex. The hemocytes of G. mellonella showed a significantly reduced ability to phagocytize the most virulent strains forming the C. haemulonii species complex. Interestingly, for C. auris, it was impossible to measure the phagocytosis index due to a general lysis of the hemocytes. Moreover, it was observed a greater capability of biofilm formation by C. auris compared to C. haemulonii species complex. In conclusion, we observed that C. auris and C. haemulonii complex have different levels of pathogenicity in the experimental models employed in the present study.
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Dermatophytosis is the most common fungal infection in cats worldwide and plays an important role in both animal and human health due to their high zoonotic potential. Effective screening is a strong preventive measure and the fungal culture is quite useful but requires full laboratorial experience and it takes a long time to obtain the result. A rapid and accurate screening test for dermatophytosis in cats is crucial for the effective control of disease outbreaks. The aim of this study was to develop and evaluate the diagnostic efficacy of enzyme immunoassays (ELISA and Western blot [WB]) for the rapid and precise diagnosis of dermatophytosis in cats. Seventy cats of various ages were divided into three groups: S (symptomatic, n = 20), AS (asymptomatic, n = 30), and N (negative, n = 20). All animals were submitted to fungal culture and blood samples for carrying out the serological tests. A significant difference (P < 0.05) was found between IgG-specific levels of sera of Microsporum canis positive and negative animals. There was no statistic difference between groups symptomatic and asymptomatic. The ELISA test showed sensitivity of 94% and specificity of 75%. Receiver operating characteristic (ROC) analysis also showed higher diagnostic accuracy (AUC 0.925). The WB technique detected 13 bands, and the 50 kDa protein was considered the most immunogenic protein, observing reactivity in 83.3% in the symptomatic group and 66.6% in the asymptomatic group. The study concluded that ELISA and WB were useful tools to reliably detect cats that have been exposed to M. canis.
Assuntos
Anticorpos Antifúngicos/sangue , Western Blotting/métodos , Doenças do Gato/diagnóstico , Ensaio de Imunoadsorção Enzimática/métodos , Microsporum/imunologia , Testes Sorológicos/métodos , Tinha/veterinária , Animais , Gatos , Imunoglobulina G/sangue , Curva ROC , Sensibilidade e Especificidade , Tinha/diagnósticoRESUMO
Paracoccidioidomycosis (PCM) is an infectious disease endemic to South America, caused by the thermally dimorphic fungi Paracoccidioides. Currently, there is no effective human vaccine that can be used in prophylactic or therapeutic regimes. We tested the hypothesis that the immunogenicity of the immunodominant CD4+ T-cell epitope (P10) of Paracoccidioides brasiliensis gp43 antigen might be significantly enhanced by using a hepatitis B virus-derived particle (VLP) as an antigen carrier. This chimera was administered to mice as a (His)6-purified protein (rPbT) or a replication-deficient human type 5 adenoviral vector (rAdPbT) in an immunoprophylaxis assay. The highly virulent Pb18 yeast strain was used to challenge our vaccine candidates. Fungal challenge evoked robust P10-specific memory CD4+ T cells secreting protective Th-1 cytokines in most groups of immunized mice. Furthermore, the highest level of fungal burden control was achieved when rAdPbT was inoculated in a homologous prime-boost regimen, with 10-fold less CFU recovering than in non-vaccinated mice. Systemic Pb18 spreading was only prevented when rAdPbT was previously inoculated. In summary, we present here VLP/P10 formulations as vaccine candidates against PCM, some of which have demonstrated for the first time their ability to prevent progression of this pernicious fungal disease, which represents a significant social burden in developing countries.
Assuntos
Antígenos de Fungos/imunologia , Linfócitos T CD4-Positivos/imunologia , Epitopos de Linfócito T/imunologia , Proteínas Fúngicas/imunologia , Vacinas Fúngicas/administração & dosagem , Glicoproteínas/imunologia , Paracoccidioides/crescimento & desenvolvimento , Paracoccidioides/imunologia , Paracoccidioidomicose/prevenção & controle , Animais , Citocinas/imunologia , Citocinas/metabolismo , Epitopos de Linfócito T/genética , Vacinas Fúngicas/imunologia , Vírus da Hepatite B/genética , Imunização , Epitopos Imunodominantes/imunologia , Imunogenicidade da Vacina , Memória Imunológica , Fígado/microbiologia , Pulmão/microbiologia , Camundongos Endogâmicos BALB C , Paracoccidioidomicose/imunologia , Paracoccidioidomicose/microbiologia , Baço/microbiologia , Células Th1/imunologia , Vacinas Sintéticas/administração & dosagem , Vacinas Sintéticas/genética , Vacinas Sintéticas/imunologia , Vacinas de Partículas Semelhantes a Vírus/administração & dosagem , Vacinas de Partículas Semelhantes a Vírus/genética , Vacinas de Partículas Semelhantes a Vírus/imunologiaRESUMO
Paracoccidioidomycosis (PCM) is an endemic systemic mycosis in Latin America, with the highest prevalence in Brazil, Colombia, and Venezuela. Fungi of the Paracoccidioides genus are etiologic agents of the disease. The 15 amino acid peptide P10 is derived from gp43, the main diagnostic antigen of Paracoccidioides brasiliensis. We previously reported that P10-pulsed dendritic cells (DCs) induce a protective response against P. brasiliensis. Presently, dexamethasone-treated BALB/c mice were intratracheally infected with P. brasiliensis Pb18 to establish the therapeutic efficacy of P10-pulsed DCs. Immunosuppressed and infected animals that received DCs had a reduction in their fungal burden, and this result was most pronounced in mice receiving DCs primed with P10. The efficacy of therapeutic DCs was significantly augmented by concomitant treatment with trimethoprim-sulfamethoxazole. Additionally, primed-DCs with or without the antifungal drug induced a beneficial Th1-biased immune response and significantly reduced tissue damage. In conclusion, these studies with immunocompromised mice demonstrate that P10-pulsed DCs with or without concomitant antifungal drugs are potently effective in combating invasive PCM. These findings support further translational studies to validate the use of P10-primed DCs for PCM in immunocompetent and immunosuppressed hosts.
RESUMO
Vulvovaginal and invasive candidiasis are frequent conditions in immunosuppressed individuals caused by Candida albicans and non-albicans Candida spp. Fluconazole and Amphotericin B are the main drugs used to fight the infection. However, resistance to fluconazole and other azole antifungal drugs is an important clinical problem that encourages the search for new therapeutic alternatives. In this work, we evaluate the antifungal activity of the biphosphinic cyclopalladate C7a in the in vitro and in vivo model. Our results showed fungicidal activity, with low values of minimal inhibitory concentrations and minimum fungicidal concentrations, even for fluconazole and/or miconazole resistant Candida isolates. Fluorescence microscopy and transmission electron microscopy revealed that the compound was able to inhibit the formation of hyphae/pseudohyphae and, moreover, promoted morphological alterations in cellular organelles and structures, such as disruption of cell wall, apparent mitochondrial swelling, chromatin marginalization into the nuclei and increased numbers of electron-lucent vacuoles. C7a significantly decreased the biofilm formation and reduced the viability of yeast cells in mature biofilms when tested against a virulent C. albicans strain. In vivo assays demonstrated a significant decrease of fungal burden in local (vaginal canal) and disseminated (kidneys) infection. In addition, we observed a significant increase in the survival of the systemically infected animals treated with C7a. Our results suggest C7a as a novel therapeutic agent for vaginal and disseminated candidiasis, and an alternative for conventional drug-resistant Candida.
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In this study, we analyzed the impact of immunization with the peptidorhamnomannan (PRM) from the cell wall of the fungus Scedosporium (Lomentospora) prolificans in a murine model of invasive scedosporiosis. Immunization with PRM decreased the survival of mice infected with S. prolificans. Immunization of mice with PRM led to decreased secretion of pro-inflammatory cytokines and chemokines but did not affect the secretion of IL-10. Mice immunized with PRM showed an increase in IgG1 secretion, which is an immunoglobulin linked to a nonprotective response. Splenocytes isolated from mice infected with S. prolificans and immunized with PRM showed no differences in the percentages of Th17 cells and no increase in the frequency of the CD4(+)CD62L(Low) T cell population. PRM-immunized mice showed a significant increase in the percentage of Treg cells. In summary, our results indicated that immunization with PRM did not assist or improve the immunological response against S. prolificans infection. PRM exacerbated the infection process by reducing the inflammatory response, thereby facilitating colonization, virulence and dissemination by the fungus.
Assuntos
Glicoproteínas/metabolismo , Imunossupressores/metabolismo , Micoses/microbiologia , Micoses/patologia , Scedosporium/crescimento & desenvolvimento , Scedosporium/imunologia , Animais , Modelos Animais de Doenças , Feminino , Vacinas Fúngicas/administração & dosagem , Vacinas Fúngicas/imunologia , Imunoglobulina G/sangue , Camundongos Endogâmicos BALB C , Linfócitos T Reguladores/imunologiaRESUMO
Paracoccidioidomycosis is a fungal disease endemic in Latin America. Polyclonal antibodies to acidic glycosphingolipids (GSLs) from Paracoccidioides brasiliensis opsonized yeast forms in vitro increasing phagocytosis and reduced the fungal burden of infected animals. Antibodies to GSL were active in both prophylactic and therapeutic protocols using a murine intratracheal infection model. Pathological examination of the lungs of animals treated with antibodies to GSL showed well-organized granulomas and minimally damaged parenchyma compared to the untreated control. Murine peritoneal macrophages activated by IFN-γ and incubated with antibodies against acidic GSLs more effectively phagocytosed and killed P. brasiliensis yeast cells as well as produced more nitric oxide compared to controls. The present work discloses a novel target of protective antibodies against P. brasiliensis adding to other well-studied mediators of the immune response to this fungus.
RESUMO
Effects on diets with reduction of crude protein and methionine + cystine (RCP) and protease supplementation (PRO) for laying hens at different ages and their influence on the internal quality of eggs at different conditions (STO) and storage times (TM) are evaluated. Four hundred and twenty eggs from commercial Hy-Line W36 laying hens collected at 32, 44 and 58 weeks of age were used. Experimental design was completely randomized in a factorial scheme 3 x 2 x 2 x 2: RCP (control, enhancement of one and two times the enzyme protease), PRO (with and without protease), STO (room temperature and refrigerated) and TM (14 and 28 days), with seven replications of one egg per experimental unit. The factorial scheme 3 x 2 x 2 was used at 58 weeks of age, because TM could not be evaluated. The percentages of egg weight loss, Haugh unit, albumen percentage, percentage and yolk index were evaluated after each storage time. RCP and PRO had no effect on internal egg quality whereas eggs under refrigeration preserved their internal quality regardless of the different storage periods.(AU)
O objetivo desse estudo foi avaliar os efeitos da redução de proteína bruta e metionina + cistina (RPB) e suplementação de protease (PRO) em dietas de poedeiras comerciais em diferentes idades, e sua influência sobre a qualidade interna de ovos armazenados em diferentes condições (ARM) e períodos de estocagem (EST). Foram utilizados 420 ovos das poedeiras comerciais da linhagem Hy-Line W36 colhidos com 32, 44 e 58 semanas de idade. O delineamento experimental foi inteiramente ao acaso em esquema fatorial 3 x 2 x 2 x 2 sendo os fatores: RPB (controle, valorização de uma e duas vezes da enzima protease), PRO (sem e com protease), ARM (ambiente e refrigerado) e EST (14 e 28 dias), com sete repetições de um ovo por unidade experimental. Com 58 semanas foi utilizado o esquema fatorial 3 x 2 x 2, pois não foi possível analisar o fator EST. Após cada período de estocagem foram avaliadas as características de perda de peso do ovo em porcentagem, Unidade Haugh, porcentagem de albúmen, porcentagem e índice de gema. A RPB e PRO não apresentaram efeitos na qualidade interna dos ovos. Ovos mantidos sob refrigeração tiveram a qualidade interna conservada, independente dos diferentes períodos de estocagem.(AU)
Assuntos
Animais , Boas Práticas de Distribuição , Óvulo , Aves Domésticas , Peptídeo HidrolasesRESUMO
Effects on diets with reduction of crude protein and methionine + cystine (RCP) and protease supplementation (PRO) for laying hens at different ages and their influence on the internal quality of eggs at different conditions (STO) and storage times (TM) are evaluated. Four hundred and twenty eggs from commercial Hy-Line W36 laying hens collected at 32, 44 and 58 weeks of age were used. Experimental design was completely randomized in a factorial scheme 3 x 2 x 2 x 2: RCP (control, enhancement of one and two times the enzyme protease), PRO (with and without protease), STO (room temperature and refrigerated) and TM (14 and 28 days), with seven replications of one egg per experimental unit. The factorial scheme 3 x 2 x 2 was used at 58 weeks of age, because TM could not be evaluated. The percentages of egg weight loss, Haugh unit, albumen percentage, percentage and yolk index were evaluated after each storage time. RCP and PRO had no effect on internal egg quality whereas eggs under refrigeration preserved their internal quality regardless of the different storage periods.
O objetivo desse estudo foi avaliar os efeitos da redução de proteína bruta e metionina + cistina (RPB) e suplementação de protease (PRO) em dietas de poedeiras comerciais em diferentes idades, e sua influência sobre a qualidade interna de ovos armazenados em diferentes condições (ARM) e períodos de estocagem (EST). Foram utilizados 420 ovos das poedeiras comerciais da linhagem Hy-Line W36 colhidos com 32, 44 e 58 semanas de idade. O delineamento experimental foi inteiramente ao acaso em esquema fatorial 3 x 2 x 2 x 2 sendo os fatores: RPB (controle, valorização de uma e duas vezes da enzima protease), PRO (sem e com protease), ARM (ambiente e refrigerado) e EST (14 e 28 dias), com sete repetições de um ovo por unidade experimental. Com 58 semanas foi utilizado o esquema fatorial 3 x 2 x 2, pois não foi possível analisar o fator EST. Após cada período de estocagem foram avaliadas as características de perda de peso do ovo em porcentagem, Unidade Haugh, porcentagem de albúmen, porcentagem e índice de gema. A RPB e PRO não apresentaram efeitos na qualidade interna dos ovos. Ovos mantidos sob refrigeração tiveram a qualidade interna conservada, independente dos diferentes períodos de estocagem.
Assuntos
Animais , Aves Domésticas , Boas Práticas de Distribuição , Óvulo , Peptídeo HidrolasesRESUMO
Paracoccidioidomycosis is a systemic granulomatous disease caused by Paracoccidioides spp. A peptide from the major diagnostic antigen gp43, named P10, induces a T-CD4(+) helper-1 immune response in mice and protects against intratracheal challenge with virulent P. brasiliensis. Previously, we evaluated the efficacy of the P10 peptide alone or combined with antifungal drugs in mice immunosuppressed and infected with virulent isolate of P. brasiliensis. In the present work, our data suggest that P10 immunization leads to an effective cellular immune response associated with an enhanced T cell proliferative response. P10-stimulated splenocytes increased nitric oxide (NO) production and induced high levels of IFN-γ, IL-1ß and IL-12. Furthermore, significantly increased concentrations of pro-inflammatory cytokines were also observed in lung homogenates of immunized mice. P10 immunization was followed by minimal fibrosis in response to infection. Combined with antifungal drugs, P10 immunization most significantly improved survival of anergic infected mice. Administration of either itraconazole or sulfamethoxazole/trimethoprim together with P10 immunization resulted in 100 % survival up to 200 days post-infection, whereas untreated mice died within 80 days. Hence, our data show that P10 immunization promotes a strong specific immune response even in immunocompromised hosts and thus P10 treatment represents a powerful adjuvant therapy to chemotherapy.
Assuntos
Antígenos de Fungos/imunologia , Vacinas Fúngicas/imunologia , Glicoproteínas/imunologia , Paracoccidioides/imunologia , Paracoccidioidomicose/prevenção & controle , Fragmentos de Peptídeos/imunologia , Animais , Antígenos de Fungos/administração & dosagem , Antígenos de Fungos/genética , Proliferação de Células , Citocinas/metabolismo , Modelos Animais de Doenças , Vacinas Fúngicas/administração & dosagem , Vacinas Fúngicas/genética , Glicoproteínas/administração & dosagem , Glicoproteínas/genética , Hospedeiro Imunocomprometido , Leucócitos Mononucleares/imunologia , Masculino , Camundongos Endogâmicos BALB C , Óxido Nítrico/metabolismo , Fragmentos de Peptídeos/administração & dosagem , Fragmentos de Peptídeos/genética , Baço/imunologia , Análise de Sobrevida , Vacinação/métodosRESUMO
Paracoccidioidomycosis is a granulomatous systemic mycosis endemic in Brazil and other Latin America countries. A DNA vaccine encoding the immunoprotective peptide 10 (P10) significantly reduced the fungal burden in mice when given prior to or after intratracheal challenge with Paracoccidioides brasiliensis. Presently, the generation/expansion of CD4+ CD44hi memory T cells as well as Foxp3+ Treg cells in mice immunized with the DNA vaccine (pcDNA3-P10) before and after infection with P. brasiliensis was investigated. Memory CD4+ CD44hi T cells simultaneously with Foxp3+ Treg cells increased in the spleens and lungs of pcDNA3-P10 immunized mice on day 0, 30, 60 and 120 postinfection. Histopathology of the lung tissue showed minimal inflammation in immunized mice compared with the unimmunized group, suggesting a role for regulatory T cells in controlling the immunopathology. The DNA vaccine shows that the repeated immunization generates memory cells and regulatory T cells that replace the initially protective pro-inflammatory T cells conferring a long term protection while preserving the integrity of the infected tissue.