RESUMO
Leptospirosis, a neglected zoonotic disease, is caused by pathogenic spirochetes belonging to the genus Leptospira and has one of the highest morbidity and mortality rates worldwide. Vaccination stands out as one of the most effective preventive measures for susceptible populations. Within the outer membrane of Leptospira spp., we find the LIC12287, LIC11711, and LIC13259 lipoproteins. These are of interest due to their surface location and potential immunogenicity. Thorough examination revealed the conservation of these proteins among pathogenic Leptospira spp.; we mapped the distribution of T- and B-cell epitopes along their sequences and assessed the 3D structures of each protein. This information aided in selecting immunodominant regions for the development of a chimeric protein. Through gene synthesis, we successfully constructed a chimeric protein, which was subsequently expressed, purified, and characterized. Hamsters were immunized with the chimeric lipoprotein, formulated with adjuvants aluminum hydroxide, EMULSIGEN®-D, Sigma Adjuvant System®, and Montanide™ ISA206VG. Another group was vaccinated with an inactivated Escherichia coli bacterin expressing the chimeric protein. Following vaccination, hamsters were challenged with a virulent L. interrogans strain. Our evaluation of the humoral immune response revealed the production of IgG antibodies, detectable 28 days after the second dose, in contrast to pre-immune samples and control groups. This demonstrates the potential of the chimeric protein to elicit a robust humoral immune response; however, no protection against challenge was achieved. While this study provides valuable insights into the subject, further research is warranted to identify protective antigens that could be utilized in the development of a leptospirosis vaccine. KEY POINTS: ⢠Several T- and B-cell epitopes were identified in all the three proteins. ⢠Four different adjuvants were used in vaccine formulations. ⢠Immunization stimulated significant levels of IgG2/3 in vaccinated animals.
Assuntos
Anticorpos Antibacterianos , Vacinas Bacterianas , Leptospirose , Lipoproteínas , Animais , Leptospirose/prevenção & controle , Leptospirose/imunologia , Lipoproteínas/imunologia , Lipoproteínas/genética , Vacinas Bacterianas/imunologia , Vacinas Bacterianas/genética , Anticorpos Antibacterianos/sangue , Anticorpos Antibacterianos/imunologia , Cricetinae , Epitopos de Linfócito B/imunologia , Epitopos de Linfócito B/genética , Proteínas Recombinantes de Fusão/imunologia , Proteínas Recombinantes de Fusão/genética , Adjuvantes Imunológicos/administração & dosagem , Imunoglobulina G/sangue , Epitopos de Linfócito T/imunologia , Epitopos de Linfócito T/genética , Leptospira interrogans/imunologia , Leptospira interrogans/genética , Proteínas da Membrana Bacteriana Externa/imunologia , Proteínas da Membrana Bacteriana Externa/genética , Vacinação , Imunidade Humoral , Leptospira/imunologia , Leptospira/genética , Imunogenicidade da VacinaRESUMO
In the last 20 years, various research groups have endeavored to develop recombinant vaccines against leptospirosis to overcome the limitations of commercially available bacterins. Numerous antigens and vaccine formulations have been tested thus far. However, the analysis of cellular response in these vaccine formulations is not commonly conducted, primarily due to the scarcity of supplies and kits for the hamster animal model. Our research group has already tested the Q1 antigen, a chimeric protein combining the immunogenic regions of LipL32, LemA, and LigANI, in recombinant subunit and BCG-vectored vaccines. In both strategies, 100 % of the hamsters were protected against clinical signs of leptospirosis. However, only the recombinant BCG-vectored vaccine provided protection against renal colonization. Thus, the objective of this study is to characterize the cellular immune response in hamsters immunized with different vaccine formulations based on the Q1 antigen through transcriptional analysis of cytokines. The hamsters were allocated into groups and vaccinated as follows: recombinant subunit (rQ1), recombinant BCG (rBCG:Q1), and saline and BCG Pasteur control vaccines. To assess the cellular response induced by the vaccines, we cultured and stimulated splenocytes, followed by RNA extraction from the cells and analysis of cytokines using real-time PCR. The results revealed that the recombinant subunit vaccine elicited a Th2-type response, characterized by the expression of cytokines IL-10, IL-1α, and TNF-α. This pattern closely resembles the cytokines expressed in severe cases of leptospirosis. On the other hand, the rBCG-vectored vaccine induced a Th1-type response with significant up-regulation of IFN-γ. These findings suggest the involvement of the cellular response and the IFN-γ mediated inflammatory response in the sterilizing immunity mediated by rBCG. Therefore, this study may assist future investigations in characterizing the cellular response in hamsters, aiming to elucidate the mechanisms of efficacy and establish potential correlates of protection.
Assuntos
Vacina BCG , Leptospirose , Cricetinae , Animais , Antígenos de Bactérias/genética , Leptospirose/prevenção & controle , Proteínas Recombinantes/genética , Vacinas Sintéticas/genética , Citocinas/metabolismo , Imunidade Celular , Proteínas Recombinantes de Fusão/genéticaRESUMO
Seroprevalence of bovine alphaherpesvirus type 1 (BoAHV1) infections may be contaminated by crossreactive antibodies to bovine alphaherpesvirus type 5 (BoAHV5). To avoid such crossreactivity, an indirect enzyme-linked immunosorbent assay prepared with a recombinant glycoprotein C (gC) antigen (ELISA-gC1) was developed, aiming the detection of antibodies to BoAHV1, with no crossreactivity with BoAHV5 antibodies. The antigen for the ELISA-gC1 was the product of the expression of 219 bp from the N-terminal portion of the BoAHV1 gC gene, which bears low homology between the two virus types. The test was validated on 131 bovine serum samples, including 26 sera from BoAHV1-experimentally immunized, 38 sera from BoAHV5-experimentally infected or immunized calves, and 67 sera from calves seronegative for both BoAHV1 and BoAHV5, as determined by serum neutralization (SN). When compared to SN for BoAHV1, the ELISA-gC1 presented 100% sensitivity, 95.5 % specificity, 100 % negative predictive value, 89.6 % positive predictive value, 98.8 % precision, and a kappa correlation coefficient (κ) 0.95. None of the 38 BoAHV5-seropositive calves was detected by the ELISA-gC1. The ELISA-gC1 proved highly effective for the identification of BoAHV1-positive sera, with no crossreactivity with anti-BoAHV5 antibodies, thus able to distinguish serological responses from BoAHV1- and BoAHV5-seropositive cattle. Its capacity to detect BoAHV1-specific antibodies should allow the determination of the actual BoAHV1 prevalence in herds, which cannot be serologically determined in countries where BoAHV5 is also prevalent due to antibody crossreactivity. Apart from recognizing exclusively BoAHV1-infected cattle, the ELISA-gC1 may also be used in support of BoAHV5 epidemiological studies by allowing the exclusion of BoAHV1-seropositive animals.
Assuntos
Doenças dos Bovinos , Herpesvirus Bovino 1 , Animais , Bovinos , Estudos Soroepidemiológicos , Anticorpos Antivirais , Ensaio de Imunoadsorção Enzimática/veterinária , Valor Preditivo dos Testes , Doenças dos Bovinos/diagnóstico , Sensibilidade e EspecificidadeRESUMO
The first leptospiral recombinant vaccine was developed in the late 1990s. Since then, progress in the fields of reverse vaccinology (RV) and structural vaccinology (SV) has significantly improved the identification of novel surface-exposed and conserved vaccine targets. However, developing recombinant vaccines for leptospirosis faces various challenges, including selecting the ideal expression platform or delivery system, assessing immunogenicity, selecting adjuvants, establishing vaccine formulation, demonstrating protective efficacy against lethal disease in homologous challenge, achieving full renal clearance using experimental models, and reproducibility of protective efficacy against heterologous challenge. In this review, we highlight the role of the expression/delivery system employed in studies based on the well-known LipL32 and leptospiral immunoglobulin-like (Lig) proteins, as well as the choice of adjuvants, as key factors to achieving the best vaccine performance in terms of protective efficacy against lethal infection and induction of sterile immunity.
RESUMO
Dogs are highly susceptible to leptospirosis and are a public health concern due to their important role as a source of spreading disease, particularly in urban settings. In this study, we present the pathogenesis, serological characterization, and complete genome sequencing of a virulent Brazilian strain (NEG7) of L. interrogans serovar Copenhageni isolated from the urine of a dog that died due to acute leptospirosis. Clinical investigation showed that the dog was presented with icteric mucous membranes, weakness, dehydration, anorexia, and kidney and liver failures. Necropsy followed by histopathological evaluation revealed lesions compatible with liver and kidney leptospirosis. The leptospires recovered from the urine were further characterized by genome analysis, which confirmed that the isolate belonged to L. interrogans serogroup icterohaemorrhagiae serovar Copenhageni. Multiple bioinformatics tools were used to characterize the genomic features, and comparisons with other available Copenhageni strains were performed. Characterization based on absence of an INDEL in the gene lic12008, associated with phylogenetic and ANI (99.99% identity) analyses, confirmed the genetic relatedness of the isolate with L. interrogans serovar Copenhageni. A better understanding of the diversity of the pathogenic Leptospira isolates could help in identifying genotypes responsible for severe infections. Moreover, it can be used to develop control and prevention strategies for Leptospira serovars associated with particular animal reservoirs.
RESUMO
Different approaches are in use to improve our knowledge about the causative agent of coronavirus disease (COVID-19). Cell culture-based methods are the better way to perform viral isolation, evaluate viral infectivity, and amplify the virus. Furthermore, next-generation sequencing (NGS) have been essential to analyze a complete genome and to describe new viral species and lineages that have arisen over time. Four naso-oropharyngeal swab samples, collected from April to July of 2020, were isolated and sequenced aiming to produce viral stocks and analyze the mutational profile of the found lineage. B.1.1.33 was the lineage detected in all sequences. Although the samples belong to the same lineage, it was possible to evaluate different mutations found including some that were first described in these sequences, like the S:H655Y and T63N. The results described here can help to elicit how the pandemic started to spread and how it has been evolving in south Brazil.
Assuntos
COVID-19 , SARS-CoV-2 , Brasil , Genoma Viral , Humanos , Mutação , Filogenia , SARS-CoV-2/genéticaRESUMO
Mycobacterium bovis BCG has been used for a century as the only licensed vaccine against tuberculosis. Owing to its strong adjuvant properties, BCG has also been employed as an oncological immunotherapeutic as well as a live vaccine vector against other pathogens. However, BCG vaccination has limited efficacy in protecting against adult forms of tuberculosis (TB), raises concerns about its safety in immunocompromised populations, compromises the diagnosis of TB through the tuberculin test and lacks predictability for successful antigen expression and immune responses to heterologous antigens. Together, these factors propelled the construction and evaluation of auxotrophic BCG strains. Auxotrophs of BCG have been developed from mutations in the genes required for their growth using different approaches and have shown the potential to provide a model to study M. tuberculosis, a more stable, safe, and effective alternative to BCG and a vector for the development of recombinant live vaccines, especially against HIV infection. In this review, we provide an overview of the strategies for developing and using the auxotrophic BCG strains in different scenarios.
RESUMO
Leptospirosis is a zoonotic disease caused by pathogenic species of Leptospira. Due to the similarity with clinical signs of other febrile diseases, early diagnosis remains challenging. Real-time PCR has been used for direct detection of Leptospira, but it requires thermocyclers and highly trained personnel. Loop-mediated isothermal amplification (LAMP) is a simple and rapid DNA-based assay. Therefore, here we have developed PCR and LAMP assays targeting two novel genes, lic13162 and lic20239, and also lipL32 gene to detect pathogenic Leptospira. Analytical and diagnostic performances were compared with bacterial isolates (including different Leptospira species and serovars) and clinical samples. The results demonstrated that PCR assays targeting lic13162 and lic20239 were successful to amplify Leptospira, but LAMP not. However, both PCR and LAMP targeting lipL32 could detect pathogenic Leptospira. LAMP lipL32 could be performed in 30 min with a detection limit of 156 cells/mL. Diagnostic performance of lipL32-LAMP presented 84.2% sensitivity and 93.2% specificity. In conclusion, lipL32 PCR and LAMP are effective methods to detect pathogenic Leptospira directly from clinical samples.
Assuntos
Leptospira , Leptospirose , Humanos , Leptospira/genética , Leptospirose/diagnóstico , Leptospirose/microbiologia , Técnicas de Diagnóstico Molecular , Técnicas de Amplificação de Ácido Nucleico , Reação em Cadeia da Polimerase em Tempo Real/métodos , Sensibilidade e EspecificidadeRESUMO
Leptospirosis is a bacterial zoonotic disease with significant impact on health all over the world. Currently, bacterins are the only vaccines available for prevention of this disease, despite several drawbacks. In an effort to develop a more effective vaccine against leptospirosis, reverse and structural vaccinology have been applied to design recombinant constructions composed of leptospiral surface-exposed antigens. Herein, we describe a protocol for design and development of Leptospirosis recombinant vaccines using immunoinformatic approaches.
Assuntos
Leptospira , Leptospirose , Antígenos de Bactérias , Vacinas Bacterianas , Humanos , Leptospira/genética , Leptospira/imunologia , Leptospirose/prevenção & controle , Vacinas Sintéticas/genéticaRESUMO
Background: Cattle are susceptible to chronic leptospirosis infection, that results in reduced milk production and reproductive disorders such as abortions, stillbirths, fetal malformation, and mummified fetuses, causing significant economic losses.Commercially available vaccines against leptospirosis offer limited protection to cattle because they contain only the mostprevalent serovars worldwide, even though they are not prevalent in the specific region. This study aimed to evaluate theprevalence of specific antibodies against Leptospira serogroups, reproductive disorders and the risk factors in dairy herdsfrom different mesoregions of Rio Grande do Sul State, Southern Brazil.Materials, Methods & Results: An epidemiological survey was conducted, and serum samples from the bovine population representative of three mesoregions (MR1, MR2, and MR3) were studied; the samples were collected and tested forleptospirosis using the microscopic agglutination test (MAT) for 12 serogroups checking for the presence of agglutination.A total of 442 blood samples were collected from dairy cattle from November to December 2019 (MR1, 187; MR2, 88;and MR3, 167), including cows vaccinated with different commercial vaccines during the three months before sample collection (n = 295) and non-vaccinated against leptospirosis (n = 147). At the time of collection, an interview was conductedwith the owners with questions about the health of the animals, management, habitat, feeding and reproduction. Chi-squaretests univariate analysis with the SPSS® version 20.0 were performed to estimate the association of serogroup Djasimanseroreactivity with the occurrence of reproductive problems and related risk factors. The mean prevalence of antibodiesagainst leptospires was 78.7% (MR1, 74.9 %; MR2, 84.1 %; and MR3, 80.2 %). Serogroup prevalence was different in...
Assuntos
Animais , Bovinos , Aborto Animal/etiologia , Fatores de Risco , Leptospirose/epidemiologia , Leptospirose/etiologia , Leptospirose/veterinária , Brasil , Inquéritos Epidemiológicos/métodos , Áreas AlagadasRESUMO
Due to the high rate of transmissibility, Brazil became the new COVID-19 outbreak epicenter and, since then, is being monitored to understand how SARS-CoV-2 mutates and spreads. We combined genomic and structural analysis to evaluate genomes isolated from different regions of Brazil and show that the most prevalent mutations were located in the S, N, ORF3a and ORF6 genes, which are involved in different stages of viral life cycle and its interaction with the host cells. Structural analysis brought to light the positions of these mutations on protein structures, contributing towards studies of selective structure-based drug discovery and vaccine development.
Assuntos
COVID-19/genética , Mutação/genética , SARS-CoV-2/genética , Proteínas Virais/genética , Brasil , Genoma Viral , Genômica , Humanos , SARS-CoV-2/patogenicidade , Índice de Gravidade de DoençaRESUMO
The occurrence of multidrug-resistant Serratia marcescens strains represents a serious public health threat. The purpose here is to report three cases of carbapenem-resistant S. marcescens infections with unfavorable clinical outcomes and provide a molecular description of the antibiotic resistance determinants at a genomic level. We performed bacterial identification by VITEK 2 and MALDI-TOF. The minimal inhibitory concentrations of antimicrobials were determined according to the Clinical and Laboratory Standards Institute guidelines, except for tigecycline, for which they were determined using Etest strips. Preliminary screening for the presence of carbapenemases was performed by ertapenem hydrolysis using MALDI-TOF MS. Whole-genome sequencing was provided to identify genes responsible for virulence and antimicrobial resistance. Here we report three challenging cases of S. marcescens that were resistant to the most commonly used antibiotics. Otherwise, we performed a genome description, which includes several genes involved in the resistance and virulence. These cases illustrate serious infection due to multidrug-resistant organisms and the complexity of treatment. Our results highlight the need to evaluate isolates regularly during long-term hospital stay to achieve optimal quality of clinical care and thus improve patient outcomes.
Assuntos
Antibacterianos , Farmacorresistência Bacteriana Múltipla , Serratia marcescens , Antibacterianos/uso terapêutico , Carbapenêmicos , Genoma Bacteriano , Humanos , Unidades de Terapia Intensiva , Testes de Sensibilidade Microbiana , Serratia marcescens/efeitos dos fármacos , Serratia marcescens/genética , Virulência , Sequenciamento Completo do GenomaRESUMO
OBJECTIVES: This study aimed to determine whether there are differences in the language used in grant applications submitted to a Southern Brazil Research Support Foundation (FAPERGS) according to the gender, career stage, and the number of publications of applicants. STUDY DESIGN AND SETTING: This observational study also evaluated the relationship between gender, career stage, curriculum, and writing characteristics. Summaries of all research proposals in the biomedical field of FAPERGS during the years of 2013 and 2014 were evaluated according to six language patterns (Positive emotions, Negative emotions, Analytic thinking, Clout, Authenticity, and Emotional tone) defined by the LIWC software. Applicant's gender, career stage, and the number of publications were also collected. RESULTS: Three hundred and forty-four (344) grant proposals met the inclusion criteria and were included in the analysis. No statistical differences were observed in the language pattern used by different gender applicants. In the language used by successful and unsuccessful applicants, we only found a small difference for clout (score 54.5 for not funded and 56.5 for funded grants). However, the principal investigators of successful applications had a significantly higher number of papers published (mean number of papers: 104 versus 58.5). CONCLUSIONS: Gender bias appears to be a more complex problem than just the type of language used; the way society is organized causes several gender biases that may be reflected throughout the women's career.
Assuntos
Pesquisa Biomédica/estatística & dados numéricos , Organização do Financiamento/estatística & dados numéricos , Pesquisadores/estatística & dados numéricos , Relatório de Pesquisa , Apoio à Pesquisa como Assunto/estatística & dados numéricos , Sexismo/estatística & dados numéricos , Redação , Adulto , Brasil , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Revisão da Pesquisa por Pares , Fatores SexuaisRESUMO
Leptospirosis has been widely reported in insular environments worldwide, characterizing a major public health threat. Although low-genetic biodiversity is expected in these regions, the introduction of domestic and synanthropic mammals may contribute to the wider diversity of leptospiral strains in insular settings. This study proposes a large-scale seroepidemiological investigation of Leptospira infection in animals from Fernando de Noronha archipelago and describes the characterization of the first leptospiral strain ever isolated from an insular setting in Brazil. A total of 1,265 blood samples from domestic (n = 682), synanthropic (n = 133) and wild (n = 450) animals were collected between 2007 and 2014, totalling 12 species. The presence of anti-Leptospira spp. antibodies was investigated by the microscopic agglutination test (MAT), and kidney samples from 20 synanthropic rodents were collected for the isolation of Leptospira spp. The leptospires recovered were further characterized by serogrouping with polyclonal antibodies, whole-genome sequencing and multilocus sequence typing (MLST). The MAT results revealed the presence of agglutinins in 90 samples (7.1%) and the most frequently found serogroup was Icterohaemorrhagiae (n = 57) in practically all species included. Viable leptospires were recovered from one brown rat, and characterization revealed that the isolate belongs to L. interrogans serogroup Pyrogenes. The results suggest that synanthropic rodents might play an important role in leptospiral infection among wildlife and domestic species in the archipelago.
Assuntos
Leptospira , Leptospirose , Doenças dos Roedores , Animais , Brasil/epidemiologia , Leptospira/genética , Leptospirose/epidemiologia , Leptospirose/veterinária , Tipagem de Sequências Multilocus/veterinária , Ratos , Doenças dos Roedores/epidemiologia , RoedoresRESUMO
Leptospirosis is a zoonotic disease worldwide and caused by the pathogenic spirochetes of the genus Leptospira. Bacterins make up the vaccines used against leptospirosis, but they only succeed in providing short-term and serovar-specific protection. The use of Mycobacterium bovis BCG as a live vaccine vector expressing leptospiral antigens is a promising alternative, particularly due to its adjuvant properties. Four distinct portions P1 (lipL32), P2 (ligAni), P3 (lemA:ligAni) and P4 (lipL32:lemA) of a recombinant chimera composed of the lipL32, lemA and ligANI genes from Leptospira interrogans were cloned individually according to the BioBricks® strategy in the plasmid pUP500/PpAN. These constructs were individually transformed into a BCG Pasteur strain, and protein expression was detected by Western blot. For vaccination, 5 groups of 10 Golden Syrian hamsters were used, aged 4-6 weeks - group 1, rBCG (LipL32); group 2, rBCG (LigAni); group 3, rBCG (LemA:LigAni); group 4, (LipL32:LemA); group 5, wild-type BCG Pasteur (negative control). Two doses containing 106 CFU of rBCG were administered subcutaneously, the challenge was performed with 5 × LD50 of Leptospira interrogans serovar Copenhageni L1-130, and the animals were observed for a 30-day period until the endpoint was reached. Humoral immunity was assessed via indirect ELISA, while renal colonisation was assessed by culture and quantitative real-time PCR. All vaccinated groups were protected against lethal challenge and renal colonisation, in comparison with negative control group (P < 0.05). Recombinant vaccines were not effective at inducing significant humoral immunity, which suggests the induction of cellular immunity - a characteristic of M. bovis BCG. In conclusion, all formulations provide 100% significant protection against leptospirosis in hamsters with no renal colonisation. The use of rBCG as a vaccine vector represents a promising alternative for the control of animal leptospirosis, allowing for protection against clinical signs of leptospirosis and renal colonisation.
Assuntos
Leptospira interrogans , Leptospira , Leptospirose , Mycobacterium bovis , Animais , Antígenos de Bactérias/genética , Vacina BCG , Vacinas Bacterianas , Cricetinae , Leptospira interrogans/genética , Leptospirose/prevenção & controle , Mycobacterium bovis/genéticaRESUMO
The first case of COVID-19 was reported in China in December 2019, and, as the virus has spread worldwide, the World Health Organization declared it a pandemic. Estimates on the number of COVID-19 cases do not reflect it real magnitude as testing is limited. Population based data on the proportion of the population with antibodies is relevant for planning public health policies. We aim to assess the prevalence of SARS-CoV-2 antibodies, presence of signs and symptoms of COVID-19, and adherence to isolation measures. A random sample comprising 133 sentinel cities from all states of the country will be selected. Three serological surveys, three weeks apart, will be conducted. The most populous municipality in each intermediate region of the country, defined by the Brazilian Institute of Geography and Statistics, was chosen as sentinel city. In each city, 25 census tracts will be selected, and 10 households will be systematically sampled in each tract, totaling 33,250 participants. In each household, one inhabitant will be randomly selected to be interviewed and tested for antibodies against SARS-CoV-2, using WONDFO SARS-CoV-2 Antibody Test. By evaluating a representative sample of Brazilian sentinel sites, this study will provide essential information for the design of health policies.
Assuntos
Betacoronavirus/isolamento & purificação , Técnicas de Laboratório Clínico , Infecções por Coronavirus/epidemiologia , Pneumonia Viral/epidemiologia , Saúde Pública , Anticorpos Antivirais/sangue , Betacoronavirus/imunologia , Brasil/epidemiologia , COVID-19 , Teste para COVID-19 , Infecções por Coronavirus/diagnóstico , Política de Saúde , Humanos , Pandemias , Pneumonia Viral/diagnóstico , Prevalência , SARS-CoV-2 , Testes SorológicosRESUMO
The aim of this study was to investigate the prevalence of thermophilic Campylobacter in the broiler production chain of southern Brazil, by evaluating broiler farms and slaughter line samples, and to determine the genetic diversity, antimicrobial resistance, and virulence genes of the isolates. Of the 140 samples investigated in this study, 75 (53.6%) were positive for thermophilic Campylobacter, and all isolates were identified by phenotypic and molecular tests as C. jejuni. The resistance to nalidixic acid was the most common (74%), followed by resistance to enrofloxacin (67.3%) and ciprofloxacin (37.1%). However, there was no resistance to the macrolides tested which are recommended for the treatment of human campylobacteriosis. The PFGE showed that the isolates were grouped in eight macrorestriction patterns (P1 to P8). A representative isolate of each macrorestriction pattern was investigated for the presence of virulence genes and all isolates carried the cadF, ciaB, cdtA, cdtB, cdtC, and flaA genes. The dnaJ gene was detected in 87.5% (7/8) of the isolates. The flhA and racR genes were detected in 75% (6/8), while the pldA gene was present in 62.5% (5/8) and the wlaN gene in 25% (2/8). The presence of C. jejuni in broiler farms and in the slaughterhouse is a hazard to consumer given that this pathogen can be maintained throughout the broiler production chain and contaminates the final product. Moreover, the presence of the major virulence genes in the isolates demonstrates that they have the ability to develop campylobacteriosis in humans.
Assuntos
Infecções por Campylobacter/veterinária , Campylobacter/genética , Galinhas/microbiologia , Farmacorresistência Bacteriana/genética , Variação Genética , Fatores de Virulência/genética , Matadouros , Animais , Antibacterianos/farmacologia , Brasil , Campylobacter/efeitos dos fármacos , Infecções por Campylobacter/microbiologia , Genes Bacterianos , Fenótipo , PrevalênciaRESUMO
Leptospirosis is a widespread zoonotic disease caused by pathogenic spirochetes of the genus Leptospira. The commercially available vaccines are bacterins that offer limited protection, short-term effect, and serovar-specific immunity. The development of novel immunization strategies is crucial to control the infection and decrease the chances of new outbreaks. In this study, purified monoclonal antibodies (mAbs) anti-LipL32 (1D9 and mAb3) were evaluated by their capacity to bind and neutralize the pathogen improving host survival. For that, an in vitro growth inhibition assay, and in vivo passive immunization were performed in animal model. Syrian hamsters were passively immunized by three different strategies. Hamsters immunized with mAb3 6 h prior to the lethal challenge showed a significantly higher survival rate of 61.1%, and a significant reduction in tissue damage in the lungs. Cumulatively, our results showed that anti-LipL32 mAbs inhibited the growth of L. interrogans in vitro, and that passive immunization offered significant protection in animal model when administered prior to infection.
Assuntos
Anticorpos Antibacterianos/imunologia , Anticorpos Monoclonais/imunologia , Proteínas da Membrana Bacteriana Externa/imunologia , Leptospira interrogans/imunologia , Leptospirose/prevenção & controle , Lipoproteínas/imunologia , Animais , Anticorpos Antibacterianos/farmacologia , Anticorpos Monoclonais/farmacologia , Cricetinae , Modelos Animais de Doenças , Feminino , Imunização , Leptospirose/microbiologia , Leptospirose/mortalidade , Leptospirose/patologia , Resultado do TratamentoRESUMO
Campylobacter jejuni is the most common bacterial cause of foodborne diarrheal disease worldwide and is among the antimicrobial resistant "priority pathogens" that pose greatest threat to public health. The genomes of two C. jejuni isolated from poultry meat sold on the retail market in Southern Brazil phenotypically characterized as multidrug-resistant (CJ100) and susceptible (CJ104) were sequenced and analyzed by bioinformatic tools. The isolates CJ100 and CJ104 showed distinct multilocus sequence types (MLST). Comparative genomic analysis revealed a large number of single nucleotide polymorphisms, rearrangements, and inversions in both genomes, in addition to virulence factors, genomic islands, prophage sequences, and insertion sequences. A circular 103-kilobase megaplasmid carrying virulence factors was identified in the genome of CJ100, in addition to resistance mechanisms to aminoglycosides, beta-lactams, macrolides, quinolones, and tetracyclines. The molecular characterization of distinct phenotypes of foodborne C. jejuni and the discovery of a novel virulence megaplasmid provide useful data for pan-genome and large-scale studies to monitor the virulent C. jejuni in poultry meat is warranted.
Assuntos
Antibacterianos/farmacologia , Campylobacter jejuni , Farmacorresistência Bacteriana Múltipla/genética , Carne/microbiologia , Animais , Brasil , Campylobacter jejuni/efeitos dos fármacos , Campylobacter jejuni/genética , Campylobacter jejuni/isolamento & purificação , Campylobacter jejuni/patogenicidade , Genoma Bacteriano/genética , Genômica , Tipagem de Sequências Multilocus , Plasmídeos/genética , Aves Domésticas , Fatores de Virulência/genéticaRESUMO
Abstract The first case of COVID-19 was reported in China in December 2019, and, as the virus has spread worldwide, the World Health Organization declared it a pandemic. Estimates on the number of COVID-19 cases do not reflect it real magnitude as testing is limited. Population based data on the proportion of the population with antibodies is relevant for planning public health policies. We aim to assess the prevalence of SARS-CoV-2 antibodies, presence of signs and symptoms of COVID-19, and adherence to isolation measures. A random sample comprising 133 sentinel cities from all states of the country will be selected. Three serological surveys, three weeks apart, will be conducted. The most populous municipality in each intermediate region of the country, defined by the Brazilian Institute of Geography and Statistics, was chosen as sentinel city. In each city, 25 census tracts will be selected, and 10 households will be systematically sampled in each tract, totaling 33,250 participants. In each household, one inhabitant will be randomly selected to be interviewed and tested for antibodies against SARS-CoV-2, using WONDFO SARS-CoV-2 Antibody Test. By evaluating a representative sample of Brazilian sentinel sites, this study will provide essential information for the design of health policies.
Resumo O COVID-19 é causado pelo vírus SARS-CoV-2, sendo o primeiro caso relatado na China em dezembro de 2019. O vírus se espalhou pelo mundo, levando a Organização Mundial da Saúde a declarar uma pandemia. As estimativas do número de casos de COVID-19 não refletem sua magnitude real, pois os testes são limitados em muitos países. Dados populacionais sobre a proporção da população com anticorpos são relevantes para o planejamento de políticas públicas de saúde. Nosso objetivo é avaliar a prevalência de anticorpos SARS-CoV-2, a presença de sinais e de sintomas de COVID-19 e a adesão a medidas de isolamento. Uma amostra aleatória composta por 133 cidades sentinelas de todos os estados do país será selecionada. Serão realizados três levantamentos sorológicos, com três semanas de intervalo. Em cada cidade, serão selecionados 25 setores censitários e 10 famílias serão amostradas aleatoriamente em cada setor. Em cada domicílio, um habitante será selecionado aleatoriamente para ser entrevistado e testado para anticorpos contra SARS-CoV-2, usando o Teste de Anticorpo WONDFO SARS-CoV-2, que foi validado antes do trabalho de campo. Ao avaliar uma amostra representativa dos locais sentinela ao longo do tempo, este estudo fornecerá informações essenciais para o desenho de políticas de saúde.