Of Animal and Men: The Importance of Animal Environment to Antimicrobial Resistance: A One Health Approach

The contribution of the animal environments to the worsening of the global antimicrobial resistance framework is related to the use of antimicrobials in subtherapeutic doses and, for long periods, establishing ideal conditions for the circulation of resistance genes, which can be transmitted to pathogens adapted to the human microbiota. The study of the animal environment as conducive to the acceleration of resistance evolution is an emerging and critical area for understanding the development and dissemination of resistance genes among the circulating bacteria. The connection between people, animals, and the environment allows us to consider antimicrobial resistance in an approach within the “One Health” concept, which provides a global strategy for expanding collaboration and interdisciplinary communication. This chapter will highlight the emergence of colistin resistance, a great challenge in antimicrobial resistance field. Also, it will focus on some agents included in the priority list of superbugs of the World Health Organization (WHO) or correlated species already identified in veterinary medicine, such as the critical superbugs; priority level 1, Carbapenem-resistant Acinetobacter baumannii, Carbapenem-resistant Pseudomonas aeruginosa, and ESBL-producing Carbapenemic-resistant Enterobacteriaceae; and the high-priority, level 2, methicillin-resistant Staphylococcus aureus (MRSA)

Comparison of the human immune responses to recombinant proteins representing three distinct surface proteins of Plasmodium vivax merozoites

Universidade Federal de São Paulo (UNIFESP), Escola Paulista de Medicina (EPM) Departamento de Microbiologia, Imunologia e ParasitologiaUniversidade Federal do Pará Departamento de PatologiaCenters for Disease Control and Prevention Division of Parasitic DiseasesInstituto Evandro ChagasUniversidade Federal de Minas Gerais Departamento de ParasitologiaUNIFESP, EPM, Depto. de Microbiologia, Imunologia e ParasitologiaSciEL

TESTE DE HODGE MODIFICADO EM ÁGAR CLED PARA TRIAGEM DE Proteus mirabilis PRODUTORES DE CARBAPENEMASE

Enterobacteriaceae produtoras de carbapenemase vêm sendo descritas em todo o mundo.  Uma detecção precisa de bactérias produtoras de carpabenemase é necessária pois esta classe de antibióticos é usada no tratamento de infecções severas. A nível laboratorial, o método fenotípico para a detecção de produtores de carbapenemase é o teste de Hodge modificado. Entretanto, algumas enterobactérias tem grande motilidade dificultando a leitura e interpretação dos resultados desta técnica. O objetivo deste estudo foi validar um meio para se obter resultados confiáveis em bactérias com grande motilidade, como é o caso de Proteus mirabilis. O meio ágar Müller-Hinton, preconizado pelo CLSI, foi comparado ao ágar CLED que demostrou ser um bom meio para análise da produção de carbapenemase em Proteus mirabilis suspeitos de produzirem esta enzima embora todos os isolados tenham sido negativos no teste

Cellular and humoral immune responses against the Plasmodium vivax MSP-1(19) malaria vaccine candidate in individuals living in an endemic area in north-eastern Amazon region of Brazil

Background: Plasmodium vivax merozoite surface protein-1 (MSP-1) is an antigen considered to be one of the leading malaria vaccine candidates. PvMSP-1 is highly immunogenic and evidences suggest that it is target for protective immunity against asexual blood stages of malaria parasites. Thus, this study aims to evaluate the acquired cellular and antibody immune responses against PvMSP-1 in individuals naturally exposed to malaria infections in a malaria-endemic area in the north-eastern Amazon region of Brazil.Methods: the study was carried out in Paragominas, Para State, in the Brazilian Amazon. Blood samples were collected from 35 individuals with uncomplicated malaria. Peripheral blood mononuclear cells were isolated and the cellular proliferation and activation was analysed in presence of 19 kDa fragment of MSP-1 (PvMSP-1(19)) and Plasmodium falciparum PSS1 crude antigen. Antibodies IgE, IgM, IgG and IgG subclass and the levels of TNF, IFN-gamma and IL-10 were measured by enzyme-linked immunosorbent assay.Results: the prevalence of activated CD4(+) was greater than CD8(+) T cells, in both ex-vivo and in 96 h culture in presence of PvMSP-1(19) and PSS1 antigen. A low proliferative response against PvMSP-1(19) and PSS1 crude antigen after 96 h culture was observed. High plasmatic levels of IFN-gamma and IL-10 as well as lower TNF levels were also detected in malaria patients. However, in the 96 h supernatant culture, the dynamics of cytokine responses differed from those depicted on plasma assays; in presence of PvMSP-1(19) stimulus, higher levels of TNF were noted in supernatant 96 h culture of malaria patient's cells while low levels of IFN-gamma and IL-10 were verified. High frequency of malaria patients presenting antibodies against PvMSP-1(19) was evidenced, regardless class or IgG subclass. PvMSP-1(19)-induced antibodies were predominantly on non-cytophilic subclasses.Conclusions: the results presented here shows that PvMSP-1(19) was able to induce a high cellular activation, leading to production of TNF and emphasizes the high immunogenicity of PvMSP-1(19) in naturally exposed individuals and, therefore, its potential as a malaria vaccine candidate.Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Instituto Oswaldo Cruz (FIOCRUZ, Brazil)Fiocruz MS, Inst Oswaldo Cruz, Lab Pesquisas Malaria, BR-21040900 Rio de Janeiro, RJ, BrazilFiocruz MS, Ctr Pesquisa Diagnost & Treinamento Malaria CPD M, Reference Ctr Malaria Extra Amazonian Reg Secreta, BR-21040900 Rio de Janeiro, RJ, BrazilFiocruz MS, Inst Oswaldo Cruz, Lab Biol Mol & Doencas Endem, BR-21040900 Rio de Janeiro, RJ, BrazilUniv São Paulo, Dept Analises Clin & Toxicol, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Microbiol Immunol & Parasitol, São Paulo, BrazilSVS, Inst Evandro Chagas, Programa Ensaios Clin Malaria, Belem, Para, BrazilUniversidade Federal de São Paulo, Dept Microbiol Immunol & Parasitol, São Paulo, BrazilWeb of Scienc

Conventional chemotherapy for acute myeloid leukemia: a Brazilian experience

CONTEXT: Young patients affected by acute myeloid leukemia (AML) achieve complete remission (CR) using conventional chemotherapy in about 55-85%. However, 30% of patients fail to achieve CR and the remission duration is often only about 12 months. More intensive treatment after CR seems to be necessary in order to maintain CR and obtain a definitive cure. In Brazil, few reports have been published on this important subject. OBJECTIVE: The aim of this study was to describe a Brazilian experience in the treatment of de novo acute myeloid leukemia (AML) in younger adult patients (age < 60 years). DESIGN: Retrospective analysis. SETTING: University Hospital, Hematology and Hemotherapy Center, State University of Campinas, Brazil. PARTICIPANTS: Newly diagnosed cases of de novo AML in the period from January 1994 to December 1998 were evaluated retrospectively, in relation to response to treatment, overall survival (OS) and disease free survival (DFS). Cases with acute promyelocytic leukemia (APL) were also included in this analysis. RESULTS: On the basis of an intention to treat, 78 cases of AML, including 17 cases of APL, were evaluated. The overall median follow-up was 272 days. The complete remission (CR) rate was 63.6% in the AML group (excluding APL) and 78% in the APL group. The 5-year estimated disease-free survival (DFS) was 80% for the APL group and 34% for the AML group (P = 0.02). The 5-year estimated overall survival (OS) was 52% for the APL group and 20.5% for the AML group, respectively (P = NS). Relapse was observed in 12/39 (30.7%) patients with AML and 1/11 (9%) with APL. CONCLUSIONS: These results are similar to those reported in the literature. However, relapse and mortality rates remain high, and a search for more aggressive strategies in order to prevent relapse is recommended.CONTEXTO: Pacientes adultos com diagnóstico de leucemia mielóide aguda (LMA) obtém remissão completa (RC) com quimioterapia convencional em cerca de 55-85% dos casos, e a sua duração é de aproximadamente 12 meses. Cerca de 30% dos pacientes não atingem a RC. Para manter a RC e obter cura definitiva parece ser necessário intensificar o tratamento após a RC. No Brasil, há um pequeno número de artigos publicados a esse respeito. OBJETIVO: Descrever a experiência brasileira no tratamento de leucemia mielóide aguda de novo em jovens e adultos com idade inferior a 60 anos. TIPO DE ESTUDO: Análise retrospectiva. LOCAL: Hospital das Clínicas e Hemocentro da Universidade Estadual de Campinas. PARTICIPANTES: Foram estudados, retrospectivamente, casos novos de LMA entre 1994 e 1998, com relação à resposta ao tratamento de quimioterapia, sobrevida global e sobrevivência livre de doença. RESULTADOS: Entraram no estudo 78 pacientes de LMA, incluindo 17 casos de leucemia promielocítica (LPM). A média de seguimento foi de 272 dias. A taxa de RC foi de 63,6% nos pacientes com LMA excluindo-se os pacientes com LPM, e 78% nos pacientes com LPM. A sobrevida livre de doença em 5 anos foi de 80% nos pacientes com LPM e 34% para os pacientes com LMA (P=0,01). A sobrevida global foi de 52% e 20,5% para os pacientes com LPM e LMA respectivamente (P=NS). Recidiva foi observada em 12/39 (30,7%) dos pacientes com LMA e 1/11 (9%) na LPM. CONCLUSÕES: Esses resultados são semelhantes aos encontrados na literatura, entretanto, o número de recidivas e a mortalidade mantém-se alta, recomendando-se pesquisa de estratégias mais agressivas para prevenir recidivas.17317

Cellular and humoral immune responses against the Plasmodium vivax MSP-119 malaria vaccine candidate in individuals living in an endemic area in north-eastern Amazon region of Brazil

Abstract\ud \ud \ud \ud Background\ud \ud Plasmodium vivax merozoite surface protein-1 (MSP-1) is an antigen considered to be one of the leading malaria vaccine candidates. PvMSP-1 is highly immunogenic and evidences suggest that it is target for protective immunity against asexual blood stages of malaria parasites. Thus, this study aims to evaluate the acquired cellular and antibody immune responses against PvMSP-1 in individuals naturally exposed to malaria infections in a malaria-endemic area in the north-eastern Amazon region of Brazil.\ud \ud \ud \ud Methods\ud The study was carried out in Paragominas, Pará State, in the Brazilian Amazon. Blood samples were collected from 35 individuals with uncomplicated malaria. Peripheral blood mononuclear cells were isolated and the cellular proliferation and activation was analysed in presence of 19 kDa fragment of MSP-1 (PvMSP-119) and Plasmodium falciparum PSS1 crude antigen. Antibodies IgE, IgM, IgG and IgG subclass and the levels of TNF, IFN-γ and IL-10 were measured by enzyme-linked immunosorbent assay.\ud \ud \ud \ud Results\ud The prevalence of activated CD4+ was greater than CD8+ T cells, in both ex-vivo and in 96 h culture in presence of PvMSP-119 and PSS1 antigen. A low proliferative response against PvMSP-119 and PSS1 crude antigen after 96 h culture was observed. High plasmatic levels of IFN-γ and IL-10 as well as lower TNF levels were also detected in malaria patients. However, in the 96 h supernatant culture, the dynamics of cytokine responses differed from those depicted on plasma assays; in presence of PvMSP-119 stimulus, higher levels of TNF were noted in supernatant 96 h culture of malaria patient’s cells while low levels of IFN-γ and IL-10 were verified. High frequency of malaria patients presenting antibodies against PvMSP-119 was evidenced, regardless class or IgG subclass.PvMSP-119-induced antibodies were predominantly on non-cytophilic subclasses.\ud \ud \ud \ud Conclusions\ud The results presented here shows that PvMSP-119 was able to induce a high cellular activation, leading to production of TNF and emphasizes the high immunogenicity of PvMSP-119 in naturally exposed individuals and, therefore, its potential as a malaria vaccine candidate.We are grateful to all patients that agreed to participate in this study for their cooperation and generous donation of blood, which made this study possible. This work was supported by Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ, Brazil), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq, Brazil), Instituto Oswaldo Cruz (FIOCRUZ, Brazil). MFFC and CTDR are recipients of a Research Productivity Fellowship from CNPq and receive a grant from FAPERJ as "Cientistas do Nosso Estado"

The influence of intestinal parasites on Plasmodium vivax-specific antibody responses to MSP-119 and AMA-1 in rural populations of the Brazilian Amazon

Abstract\ud \ud Background\ud Polyparasitism is a common condition in humans but its impact on the host immune system and clinical diseases is still poorly understood. There are few studies of the prevalence and the effect of malaria-intestinal parasite co-infections in the immune response to malaria vaccine candidates. The present study determines whether the presence of malaria and intestinal parasites co-infection is associated with impaired IgG responses to Plasmodium vivax AMA-1 and MSP-119 in a rural population of the Brazilian Amazon.\ud \ud \ud Methods\ud A cross-sectional survey was performed in a rural area of Rondonia State and 279 individuals were included in the present study. At recruitment, whole blood was collected and Plasmodium and intestinal parasites were detected by microscopy and molecular tests. Blood cell count and haemoglobin were also tested and antibody response specific to P. vivax AMA-1 and MSP-119 was measured in plasma by ELISA. The participants were grouped according to their infection status: singly infected with Plasmodium (M); co-infected with Plasmodium and intestinal parasites (CI); singly infected with intestinal parasites (IP) and negative (N) for both malaria and intestinal parasites.\ud \ud \ud Results\ud The prevalence of intestinal parasites was significantly higher in individuals with malaria and protozoan infections were more prevalent. IgG antibodies to PvAMA-1 and/or PvMSP-119 were detected in 74 % of the population. The prevalence of specific IgG was similar for both proteins in all four groups and among the groups the lowest prevalence was in IP group. The cytophilic sub-classes IgG1 and IgG3 were predominant in all groups for PvAMA-1 and IgG1, IgG3 and IgG4 for PvMSP-119. In the case of non-cytophilic antibodies to PvAMA-1, IgG2 was significantly higher in IP and N group when compared to M and CI while IgG4 was higher in IP group.\ud \ud \ud Conclusions\ud The presence of intestinal parasites, mainly protozoans, in malaria co-infected individuals does not seem to alter the antibody immune responses to P. vivax AMA-1 and MSP-119. However, IgG response to both AMA1 and MSP1 were lower in individuals with intestinal parasites.The authors are in debt to the individuals who participated in this study, the\ud Secretary of Health and Laboratory Central (LACEN) of Rondonia, the local\ud malaria control team in Joana D´Arc settlement for their logistic support\ud and the Institute Oswaldo Cruz (Fiocruz) for overall support. This work was\ud supported by PRONEX Malaria network funded by the Brazilian Ministry of\ud Science and Technology (MCT), Conselho Nacional de Desenvolvimento\ud Cientifico e Tecnologico (CNPq, Brazil) and Fundação de Amparo à Pesquisa\ud do Estado do Rio de Janeiro (FAPERJ, Brazil). PROEP, Instituto Oswaldo Cruz\ud (FIOCRUZ, Brazil). JOF is recipient of a Research Productivity Fellowship from\ud CNPq, JCSA is recipient of a fellowship from Instituto Oswaldo Cruz and VAR,\ud MM from CNPq

Adaptive Immunity against Leishmania Nucleoside Hydrolase Maps Its C-Terminal Domain as the Target of the CD4+ T Cell–Driven Protective Response

Nucleoside hydrolases (NHs) show homology among parasite protozoa, fungi and bacteria. They are vital protagonists in the establishment of early infection and, therefore, are excellent candidates for the pathogen recognition by adaptive immune responses. Immune protection against NHs would prevent disease at the early infection of several pathogens. We have identified the domain of the NH of L. donovani (NH36) responsible for its immunogenicity and protective efficacy against murine visceral leishmaniasis (VL). Using recombinant generated peptides covering the whole NH36 sequence and saponin we demonstrate that protection against L. chagasi is related to its C-terminal domain (amino-acids 199–314) and is mediated mainly by a CD4+ T cell driven response with a lower contribution of CD8+ T cells. Immunization with this peptide exceeds in 36.73±12.33% the protective response induced by the cognate NH36 protein. Increases in IgM, IgG2a, IgG1 and IgG2b antibodies, CD4+ T cell proportions, IFN-γ secretion, ratios of IFN-γ/IL-10 producing CD4+ and CD8+ T cells and percents of antibody binding inhibition by synthetic predicted epitopes were detected in F3 vaccinated mice. The increases in DTH and in ratios of TNFα/IL-10 CD4+ producing cells were however the strong correlates of protection which was confirmed by in vivo depletion with monoclonal antibodies, algorithm predicted CD4 and CD8 epitopes and a pronounced decrease in parasite load (90.5–88.23%; p = 0.011) that was long-lasting. No decrease in parasite load was detected after vaccination with the N-domain of NH36, in spite of the induction of IFN-γ/IL-10 expression by CD4+ T cells after challenge. Both peptides reduced the size of footpad lesions, but only the C-domain reduced the parasite load of mice challenged with L. amazonensis. The identification of the target of the immune response to NH36 represents a basis for the rationale development of a bivalent vaccine against leishmaniasis and for multivalent vaccines against NHs-dependent pathogens