Cláudio Mafra

Detection of a typhus group Rickettsia in Amblyomma ticks in the state of Nuevo Leon, Mexico.

Abstract: The state of Nuevo Leon, Mexico has had outbreaks of typhus group rickettsiosis, most recently recognized in 1997. Evaluation of the sera of 345 patients with a dengue‐like illness revealed that 25.5% had antibodies reactive with typhus group rickettsiae and 16% had antibodies to Rickettsia parkeri. Rickettsiae were detected by PCR and shell‐vial isolations in the field‐collected Amblyomma ticks. Molecular characterization by DNA sequence analysis of the gltA, ompB, and 17‐kDa gene identified the organisms to be R. prowazekii.

Caracterização de Rickettsia spp. circulante em foco silencioso de febre maculosa brasileira no Município de Caratinga, Minas Gerais, Brasil

The present study was intended to characterize Rickettsia spp. circulating in arthropod vectors in Caratinga, Minas Gerais, Brazil, by PCR and to investigate the presence of antibodies against the spotted fever Rickettsiae group (SFRG) in dogs and horses. 2,610 arthropods were collected and taxonomically identified. DNA samples obtained from these vectors were submitted to PCR and cycle-sequenced. Ctenocephalides and Amblyomma cajennense showed sequences presenting 100.0% homology with R. felis. A sequence obtained from Rhipicephalus sanguineus showed 99.0% homology with R. felis, and a sequence from A. cajennense showed 97.0% homology with R. honei and R. rickettsii. Canine (73) and equine (18) serum samples were tested by indirect fluorescent antibody (IFA) using R. rickettsii antigen. Only three of the equine sera tested (17.0%) had positive antibody titers. Molecular detection of rickettsiae species potentially pathogenic to humans in arthropod vectors and the presence of seroreactivity to SFRG in horses show the risk of transmission of rickettsiosis in this area and the need to maintain continuous epidemiological surveillance for rickettsial diseases.

The sialotranscriptome of Amblyomma triste, Amblyomma parvum and Amblyomma cajennense ticks, uncovered by 454-based RNA-seq

BackgroundTick salivary constituents antagonize inflammatory, immune and hemostatic host responses, favoring tick blood feeding and the establishment of tick-borne pathogens in hosts during hematophagy. Amblyomma triste, A. cajennense and A. parvum ticks are very important in veterinary and human health because they are vectors of the etiological agents for several diseases. Insights into the tick salivary components involved in blood feeding are essential to understanding vector-pathogen-host interactions, and transcriptional profiling of salivary glands is a powerful tool to do so. Here, we functionally annotated the sialotranscriptomes of these three Amblyomma species, which allowed comparisons between these and other hematophagous arthropod species.MethodsmRNA from the salivary glands of A. triste, A. cajennense and A. parvum ticks fed on different host species were pyrosequenced on a 454-Roche platform to generate four A. triste (nymphs fed on guinea pigs and females fed on dogs) libraries, one A. cajennense (females fed on rabbits) library and one was A. parvum (females fed on dogs) library. Bioinformatic analyses used in-house programs with a customized pipeline employing standard assembly and alignment algorithms, protein databases and protein servers.ResultsEach library yielded an average of 100,000 reads, which were assembled to obtain contigs of coding sequences (CDSs). The sialotranscriptome analyses of A. triste, A. cajennense and A. parvum ticks produced 11,240, 4,604 and 3,796 CDSs, respectively. These CDSs were classified into over 100 distinct protein families with a wide range of putative functions involved in physiological and blood feeding processes and were catalogued in annotated, hyperlinked spreadsheets. We highlighted the putative transcripts encoding saliva components with critical roles during parasitism, such as anticoagulants, immunosuppressants and anti-inflammatory molecules. The salivary content underwent changes in the abundance and repertoire of many transcripts, which depended on the tick and host species.ConclusionsThe annotated sialotranscriptomes described herein richly expand the biological knowledge of these three Amblyomma species. These comprehensive databases will be useful for the characterization of salivary proteins and can be applied to control ticks and tick-borne diseases.

First report of Ehrlichia ewingii detected by molecular investigation in dogs from Brazil

Laboratorio de Bioquimica e Biologia Molecular de Agentes Infecciosos e Parasitarios, Departamento de Bioquimica e Biologia Molecular, Universidade Federal de Vicosa, Vicosa, Minas Gerais, Brazil, Federal University of Switzerland, Zurich, Switzerland, Laboratorio de Infectologia Molecular Animal, Departamento de Bioquimica e Biologia Molecular, Universidade Federal de Vicosa, Vicosa, Minas Gerais, Brazil, Hospital Veterinario, Departamento de Medicina Veterinaria, Universidade Federal de Vicosa, Vicosa, Minas Gerais, Brazil and Departamento de Nutricao e Clinica Social, Universidade Federal de Ouro Preto, Ouro Preto, Minas Gerais, Brazil

Tripping over emerging pathogens around the world: A phylogeographical approach for determining the epidemiology of Porcine circovirus-2 (PCV-2), considering global trading

Porcine circovirus-2 (PCV-2) is an emerging virus associated with a number of different syndromes in pigs known as Porcine Circovirus Associated Diseases (PCVAD). Since its identification and characterization in the early 1990s, PCV-2 has achieved a worldwide distribution, becoming endemic in most pig-producing countries, and is currently considered as the main cause of losses on pig farms. In this study, we analyzed the main routes of the spread of PCV-2 between pig-producing countries using phylogenetic and phylogeographical approaches. A search for PCV-2 genome sequences in GenBank was performed, and the 420 PCV-2 sequences obtained were grouped into haplotypes (group of sequences that showed 100% identity), based on the infinite sites model of genome evolution. A phylogenetic hypothesis was inferred by Bayesian Inference for the classification of viral strains and a haplotype network was constructed by Median Joining to predict the geographical distribution of and genealogical relationships between haplotypes. In order to establish an epidemiological and economic context in these analyses, we considered all information about PCV-2 sequences available in GenBank, including papers published on viral isolation, and live pig trading statistics available on the UN Comtrade database (http://comtrade.un.org/). In these analyses, we identified a strong correlation between the means of PCV-2 dispersal predicted by the haplotype network and the statistics on the international trading of live pigs. This correlation provides a new perspective on the epidemiology of PCV-2, highlighting the importance of the movement of animals around the world in the emergence of new pathogens, and showing the need for effective sanitary barriers when trading live animals.

Amblyomma imitator ticks as vectors of Rickettsia rickettsii, Mexico.

Real-time PCR of Amblyomma imitator tick egg masses obtained in Nuevo Leon State, Mexico, identified a Rickettsia species. Sequence analyses of 17-kD common antigen and outer membrane protein A and B gene fragments showed to it to be R. rickettsii, which suggested a potential new vector for this bacterium.

Clinical and laboratorial evidence of Rickettsia felis infections in Latin America

After the discovery and initial characterization of Rickettsia felis in 1992 by Azad and cols, and the subsequent first description of a human case of infection in 1994, there have been two communications of human rickettsiosis cases caused by Rickettsia felis in Latin America. The first one was published in 2000 by Zavala-Velazquez and cols in Mexico. In 2001 Raoult and cols described the occurrence of two human cases of Rickettsia felis rickettsiosis in Brazil. In the present discussion these two articles were compared and after the description of the principal signs and symptoms, it was concluded that more studies are needed with descriptions of a greater number of patients to establish the true frequency of the clinical signs and symptoms present in Rickettsia felis rickettsiosis.

Exoantigens of an attenuated strain of Babesia bovis used as a vaccine against bovine babesiosis

Bovine babesiosis caused by Babesia bovis remains a significant constraint to beef and milk cattle production throughout the world. Exoantigens released by the parasites in culture supernatants are a potential source of antigen to induce protective immunity. An attenuated strain of B. bovis from Brazil, catalogued as BbUFV1, was maintained in vitro by the MASP method, and exoantigen-containing supernatant fluids were collected daily to form a pool representing a 72-h culture cycle for preparation of the vaccine. Exoantigen concentration was estimated using a two-site EIA. Three groups of susceptible non-splenectomised male Bos taurus cattle, 14 months old, were used. Group A (vaccinated) received two subcutaneous immunizations with a 21-day interval of B. bovis supernatant, content 6500 EIA units of exoantigens plus 1.5 mg saponin, and Group B (adjuvant control) received two injections of adjuvant alone. Four weeks after the second immunization, Groups A, B and C (control) were challenged intravenously with 10(8) virulent parasites of a heterologous B. bovis strain. The results showed that exoantigens present in in vitro cultures can induce a high degree of protection against virulent heterologous challenge exposure. In Group A only one animal showed discrete parasitaemia; all developed a fever and slight decreases in PCV, with a rapid return to normal values. One animal of Group B died; the survivors showed fever, anaemia and parasitaemia. All animals of Group C died between 7 and 13 days after challenge. Vaccination elicited both humoral and cell-mediated immune responses. In Group A, after the challenge, the maximum antibody titer was 12,800. When vaccinated, cattle were tested at the moment of challenge for B. bovis-specific cell-mediated immunity by the monocytemigration inhibition test. A mean inhibition index of 60 +/- 0.33 was observed. Preliminary Western blot analysis of the immunogen revealed at least four proteins of molecular weight ranging between 30 and 160 kDa.

Molecular detection of Ehrlichia canis in cats in Brazil

Laboratorio de Bioquimica e Biologia Molecular de Agentes Infecciosos e Parasitarios, Departamento de Bioquimica e Biologia Molecular, Universidade Federal de Vicosa, Vicosa, Hospital Veterinario, Departamento de Medicina Veterinaria, Universidade Federal de Vicosa, Vicosa, Laboratorio de Infectologia Molecular Animal, Departamento de Bioquimica e Biologia Molecular, Universidade Federal de Vicosa, Vicosa and Departamento de Nutricao e Clinica Social, Universidade Federal de Ouro Preto, Ouro Preto, Minas Gerais, Brazil

Rickettsia felis in the Americas

Abstract:  The authors describe their work in the Americas in Rickettsia felis cases in humans and the presence of Rickettsia felis in vectors.