Genetic diversity of Hepatozoon spp. in rodents from Brazil.

Hepatozoon spp. are Apicomplexan protozoa that parasitize a wide diversity of vertebrate hosts. In Brazil, few studies have reported the occurrence of Hepatozoon spp. in rodent species. Additionally, an evaluation of the population structure and distribution of Hepatozoon species over several Brazilian biomes has not yet been performed. The present work aimed to investigate the genetic diversity of Hepatozoon spp. in rodents from 31 genera sampled in five Brazilian biomes. Samples were submitted to PCR assays for Hepatozoon spp. targeting two regions of the 18S rRNA gene. Infection by Hepatozoon spp. was detected in 195 (42.2%) rodents comprising 24 genera. Phylogenetic analyses of 18S rRNA sequences grouped all sequences in the clade of Hepatozoon spp. previously detected in rodents and reptiles, apart from those detected in domestic/wild carnivores. These data raise two non-exclusive hypotheses: (i) rodents play an important role as intermediate or paratenic hosts for Hepatozoon infections in reptiles; and (ii) rodents do not seem to participate in the epidemiology of Hepatozoon infections of domestic/wild canids and felids in Brazil. TCS analyses performed with available 18S rRNA Hepatozoon sequences detected in rodents from Brazil showed the occurrence of six haplotypes, which were distributed in two large groups: one from rodents inhabiting the coastal region of Brazil and Mato Grosso state, and another from rodents from the central region of the country. A wide survey of the South American territory will help to elucidate the evolutionary history of Hepatozoon spp. parasitizing Rodentia in the American continent.

Microorganisms in the ticks Amblyomma dissimile Koch 1844 and Amblyomma rotundatum Koch 1844 collected from snakes in Brazil.

Knowledge about ticks (Acari) and screening of ticks parasitizing various hosts are necessary to understand the epidemiology of tick-borne pathogens. The objective of this study was to investigate tick infestations on snakes (Reptilia: Squamata: Serpentes) arriving at the serpentarium at the Institute Vital Brazil, Rio de Janeiro. Some of the identified ticks were individually tested for the presence of bacteria of the genera Rickettsia (Rickettsiales: Rickettsiaceae), Borrelia (Spirochaetales: Spirochaetaceae), Coxiella (Legionellales: Coxiellaceae), Bartonella (Rhizobiales: Bartonellaceae), Ehrlichia (Rickettsiales: Anaplasmataceae), Anaplasma (Rickettsiales: Anaplasmataceae), and Apicomplexa protozoa of the genera Babesia (Piroplasmida: Babesiidae) and Hepatozoon (Eucoccidiorida: Hepatozoidae). A total of 115 hard ticks (Ixodida: Ixodidae) were collected from 17 host individuals obtained from four Brazilian states. Two species of tick were identified: Amblyomma dissimile Koch 1844 (four larvae, 16 nymphs, 40 adults), and Amblyomma rotundatum Koch 1844 (12 nymphs, 43 adults). Rickettsia bellii was found in A. rotundatum and A. dissimile ticks and Rickettsia sp. strain Colombianensi, Anaplasma-like and Hepatozoon sp. in A. dissimile ticks. Among the tested ticks, no DNA of Borrelia, Bartonella, Coxiella or Babesia was found. The present findings extend the geographic range of Rickettsia sp. strain Colombianensi in Brazil and provide novel tick-host associations.

Hantavirus pulmonary syndrome in a highly endemic area of Brazil.

Hantavirus pulmonary syndrome (HPS) is the most frequently reported fatal rodent-borne disease in Brazil, with the majority of cases occurring in Santa Catarina. We analysed the clinical, laboratory and epidemiological data of the 251 confirmed cases of HPS in Santa Catarina in 1999-2011. The number of cases ranged from 10 to 47 per year, with the highest incidences in 2004-2006. Gastrointestinal tract manifestations were found in >60% of the cases, potentially confounding diagnosis and leading to inappropriate therapy. Dyspnoea, acute respiratory failure, renal failure, increased serum creatinine and urea levels, increased haematocrits and the presence of pulmonary interstitial infiltrate were significantly more common in HPS patients who died. In addition, we demonstrated that the six cases from the midwest region of the state were associated with Juquitiba virus genotype. The case-fatality rate in this region, 19·2%, was lower than that recorded for other mesoregions. In the multivariate analysis increase of serum creatinine and urea was associated with death by HPS. Our findings help elucidate the epidemiology of HPS in Brazil, where mast seeding of bamboo can trigger rodent population eruptions and subsequent human HPS outbreaks. We also emphasize the need for molecular confirmation of the hantavirus genotype of human cases for a better understanding of the mortality-related factors associated with HPS cases in Brazil.

Hantavirus pulmonary syndrome and rodent reservoirs in the savanna-like biome of Brazil's southeastern region.

This paper describes the diversity of rodent fauna in an area endemic for hantavirus cardiopulmonary syndrome (HCPS) in Brazil, the population dynamics and the relationship of rodents with hantavirus in the Cerrado (savanna-like) biome. Additionally, an analysis is made of the partial S segment sequences of the hantaviruses obtained from serologically confirmed human HCPS cases and from rodent specimens. Rodents were collected during four campaigns. Human serum samples were collected from suspected cases of HCPS at hospitals in the state of Minas Gerais. The samples antibody-reactive by ELISA were processed by RT-PCR. The PCR product was amplified and sequenced. Hantavirus was detected only in Necromys lasiurus, the wild rodent species most prevalent in the Cerrado biome (min-max: 50-83·7%). All the six human serum samples were hantavirus seropositive and five showed amplified PCR products. The analysis of the nucleotide sequences showed the circulation of a single genotype, the Araraquara hantavirus. The environmental changes that have occurred in the Cerrado biome in recent decades have favoured N. lasiurus in interspecific competition of habitats, thus increasing the risk of contact between humans and rodent species infected with hantavirus. Our data corroborate the definition of N. lasiurus as the main hantavirus reservoir in the Cerrado biome.

A cluster of Rickettsia rickettsii infection at an animal shelter in an urban area of Brazil.

Rickettsia rickettsii infection is being increasingly recognized as an important cause of fatal acute illness in Brazil, where this tick-borne disease is designated Brazilian spotted fever (BSF). In this study we report five fatal cases of BSF in employees of an animal shelter in an urban area in the municipality of Rio de Janeiro in southeast Brazil after a natural disaster on 11 January 2011. Four of the cases occurred from 27 January to 11 April 2011, while the fifth fatal case was identified in April 2012. Three cases were confirmed by molecular analysis and two by epidemiological linkage. An investigation of BSF was performed in the animal shelter, and blood samples were collected from 115 employees and 117 randomly selected dogs. The presence of high levels (1024-4096) of antibodies against spotted fever group rickettsiae was found in three (2·6%) employees and 114 (97·5%) dogs. These findings emphasize the need to consider BSF as a possible cause of undifferentiated febrile illness, especially dengue and leptospirosis, in patients occupationally exposed to dogs heavily infested by ticks, mainly working at kennels and animal shelters that have inadequate space for the animals housed and frequently providing an environment conducive to exposure to pathogens such as R. rickettsii.

Regional variations and time trends of hantavirus pulmonary syndrome in Brazil.

Hantavirus pulmonary syndrome (HPS) is a zoonosis and a public health concern in the Americas due to its high fatality rate. Since the first cases were identified in USA in 1993, HPS has been observed over a wide area of the American continents. Over 1600 HPS cases have been confirmed in Brazil since 1993, and different genotypes of hantavirus have been identified in several biomes in this country of continental dimensions. We performed a retrospective study of data from 2001 to 2011 that encompassed all notified cases of HPS in the National Disease Notification System (SINAN) of Brazil to determine the regional differences and temporal trends of the disease. During the study period, 1486 cases were reported, and analyses of the temporal trends of the disease revealed that the number of cases in Brazil increased over this period (P < 0·01). The frequency distributions of the cases stratified by region revealed different patterns of seasonality; however, these patterns were not statistically significant with the exception of the South region (P < 0·05). We found regional differences in the occurrence of the disease throughout the year and a trend towards an increasing number of cases. These findings reinforce the necessity of adjusting surveillance tools and control policies to respond effectively to this public health problem.

Genetic characterization of a Juquitiba-like viral lineage in Oligoryzomys nigripes in Rio de Janeiro, Brazil.

Hantaviruses, family Bunyaviridae, are rodent-borne RNA viruses that have caused cases of hantavirus cardiopulmonary syndrome (HCPS) in various regions of the Americas. There are five hantaviral lineages associated with HCPS in Brazil: Juquitiba virus (JUQV), Araraquara virus (ARAV), Laguna Negra-like virus (LNV), Castelo dos Sonhos virus (CASV), and Anajatuba virus (ANAJV). Three additional hantaviruses have been described in rodents alone: Rio Mearim virus, Jaborá virus, and a hantavirus lineage related to Seoul virus. This study describes the genetic detection and characterization of a Juquitiba-like hantavirus in Oligoryzomys nigripes, or the black-footed pygmy rice rat, in the Serra dos Orgãos National Park, Rio de Janeiro State, where so far no cases of HCPS have been reported.

Human bartonellosis: seroepidemiological and clinical features with an emphasis on data from Brazil - A review

Bartonellae are fastidious Gram-negative bacteria that are widespread in nature with several animal reservoirs (mainly cats, dogs, and rodents) and insect vectors (mainly fleas, sandflies, and human lice). Thirteen species or subspecies of Bartonella have been recognized as agents causing human disease, including B. bacilliformis, B. quintana, B. vinsonii berkhoffii, B. henselae, B. elizabethae, B. grahamii, B. washoensis, B. koehlerae, B. rocha-limaea, and B. tamiae. The clinical spectrum of infection includes lymphadenopathy, fever of unknown origin, endocarditis, neurological and ophthalmological syndromes, Carrion's disease, and others. This review provides updated information on clinical manifestations and seroepidemiological studies with an emphasis on data available from Brazil.

Bartonella native valve endocarditis: the first brazilian case alive and well

Bartonella is an important cause of blood culture-negative endocarditis in recent studies. Seroprevalence studies in the States of Minas Gerais and Rio de Janeiro have shown Bartonella IgG positivity around 14 percent in healthy adults and 40 percent in HIV seropositive adults, respectively. A case report of a 46-year-old white male with moderate aortic regurgitation (AR) due to rheumatic heart disease (RHD), admitted due to worsening heart failure, is presented. Clinical features were apyrexia, anemia, polyclonal hypergammaglobulinemia, hematuria and splenomegaly. He was submitted to surgery due to worsening AR. Histopathology of the excised valve showed active bacterial endocarditis and underlying RHD. Routine blood cultures were negative. Indirect immunofluorescence (IFI) assays for Coxiella burnetii were non-reactive. Bartonella henselae IgG titer was 1:4096 prior to antibiotics and 1:512 14 months after treatment. History of close contact with a young cat during the months preceding his admission was elicited.

Bartonella native valve endocarditis: the first Brazilian case alive and well.

Bartonella is an important cause of blood culture-negative endocarditis in recent studies. Seroprevalence studies in the States of Minas Gerais and Rio de Janeiro have shown Bartonella IgG positivity around 14% in healthy adults and 40% in HIV seropositive adults, respectively. A case report of a 46-year-old white male with moderate aortic regurgitation (AR) due to rheumatic heart disease (RHD), admitted due to worsening heart failure, is presented. Clinical features were apyrexia, anemia, polyclonal hypergammaglobulinemia, hematuria and splenomegaly. He was submitted to surgery due to worsening AR. Histopathology of the excised valve showed active bacterial endocarditis and underlying RHD. Routine blood cultures were negative. Indirect immunofluorescence (IFI) assays for Coxiella burnetii were non-reactive. Bartonella henselae IgG titer was 1:4096 prior to antibiotics and 1:512 14 months after treatment. History of close contact with a young cat during the months preceding his admission was elicited.

Study of hantavirus infection in captive breed colonies of wild rodents.

Wild sigmondontine rodents are known to be the reservoir of several serotypes of New World hantaviruses. The mechanism of viral transmission is by aerosol inhalation of the excreta from infected rodents. Considering that the captive breed colonies of various wild mammals may present a potential risk for hantaviral transmission, we examined 85 specimens of Thrichomys spp. (Echimyidae) and 17 speciemens of Nectomys squamipes (Sigmodontinae) from our colony for the presence of hantavirus infections. Blood samples were assayed for the presence of antibodies to Andes nucleocapsid antigen using enzyme-linked immunosorbent assay (ELISA). Additionally, serum samples from workers previously exposed to wild rodents, in the laboratories where the study was conducted, were also tested by ELISA to investigate prevalence of anti-hantavirus IgG antibodies. All blood samples were negative for hantavirus antibodies. Although these results suggest that those rodent's colonies are hantavirus free, the work emphasizes the need for hantavirus serological monitoring in wild colonized rodents and secure handling potentially infected rodents as important biosafety measures.

Study of hantavirus infection in captive breed colonies of wild rodents

Wild sigmondontine rodents are known to be the reservoir of several serotypes of New World hantaviruses. The mechanism of viral transmission is by aerosol inhalation of the excreta from infected rodents. Considering that the captive breed colonies of various wild mammals may present a potencial risk for hantaviral transmission, we examined 85 speciemens of Thrichomys spp. (Echimyidae) and 17 speciemens of Nectomys squamipes (Sigmodontinae) from our colony for the presence of hantavirus infections. Blood samples were assayed for the presence of antibodies to Andes nucleocapsid antigen using enzyme-linked immunosorbent assay (ELISA). Additionally, serum samples from workers previously exposed to wild rodents, in the laboratories where the study was conducted, were also tested by ELISA to investigate prevalence of anti-hantavirus IgG antibodies. All blood samples were negative for hantavirus antibodies. Although these results suggest that those rodent's colonies are hantavirus free, the work emphasizes the need for hantavirus serological monitoring in wild colonized rodents and secure handling potentially infected rodents as important biosafety measures.

Spotted fever in Brazil: a seroepidemiological study and description of clinical cases in an endemic area in the state of São Paulo.

During 1985-1995, illnesses clinically and epidemiologically compatible with Brazilian spotted fever were identified in 17 patients in the county of Pedreira, in the state of São Paulo, Brazil. Spotted-fever group rickettsial infection was confirmed by serology and/or immunostaining of tissues in 10 of these patients. Immunostaining confirmed infection in a 37-year-old pregnant patient, although rickettsial antigens were not demonstrable in the tissues of the fetus. A serosurvey was conducted in four localities in the county to determine the prevalence of subclinical or asymptomatic infections with spotted fever group rickettsiae. Five hundred and twenty-five blood samples were tested by an indirect immunofluorescence assay for antibodies reactive with Rickettsia rickettsii. Twenty-two (4.2%) of these samples demonstrated titers > or = 1:64. The results indicate that Brazilian spotted fever is endemic within this region of Brazil.

Detection of poxvirus in cattle associated with human cases in the State of Rio de Janeiro: preliminary report.

This preliminary report describes human and cow cases of poxvirus that recently occurred in the State of Rio de Janeiro. The electron microscopic findings were consistent with parapoxviral and orthopoxviral infection. Orthopoxvirus strains were isolated from human and cow cases. Detailed viral characterization by means of genetical techniques is under investigation. Based on these informations, poxviral diseases should be also considered an emerging viral zoonosis that can affect human beings.

[Isolation of Rickettsia in vero cell culture]. / Isolamento de Rickettsia em cultura de células vero.

The diagnosis of spotted fever is based on characteristic signs and symptoms but requires laboratorial confirmation because of the possible differential diagnosis from other diseases like leptospirosis, enterovirus, meningococcemia and typhoid fever. Laboratorial confirmation may be done by detection of specific antibodies which is possible only 5-10 days after the onset of the symptoms or by the isolation of Rickettsiae from blood and/or skin biopsy and from ticks collected in the patient or in the animal reservoir. The isolation of Rickettsiae from blood or skin biopsy results in an early diagnosis of spotted fever since in the rickettsiemic phase of the disease there is no detectable level of antibodies in the serum. With the purpose of facilitating the diagnosis of Spotted Fever we have standardized the isolation of Rickettsiae in cell culture by a method that is less time consuming and that reduces the biological risks than isolation in guinea pigs. Vero cell cultures were inoculated with the Sheyla Smith strain of Rickettsia rickettsii provided by CDC (Atlanta-USA). The identification was performed by indirect immunofluorescence technique. The presence of green fluorescent organisms characterized the growth of the agent. Ulterior confirmation of the methodology was done by isolation of the spotted fever agent from skin biopsy of a patient from an endemic area and from Amblyomma ticks that are the reservoir and vector of the Brazilian spotted fever.

Age-specific prevalence of antibodies to hepatitis A in children and adolescents from Rio de Janeiro, Brazil, 1978 and 1995. Relationship of prevalence to environmental factors.

The age-specific prevalence of antibodies to hepatitis A virus (anti-HAV) was determined in two different population groups with low socio-economic status from Rio de Janeiro city, Brazil, whose serum samples were collected 17 years apart (Population 1, 1978; Population 2, 1995). In Population 2, analysis of the anti-HAV prevalence was also carried out with respect to environmental factors. Population 1 was composed of 520 stored sera collected from the umbilical cord of term neonates and children aged 1 month to 6 years. In population 2, 720 serum samples were collected from children and adolescents with ages ranging from 1 to 23 years. The overall prevalence rate of anti-HAV in Population 1 and Population 2 was 65.6% and 32.1%, respectively. In Population 1, the anti-HAV prevalence reached 88% at the age of 3, while in Population 2, it increased from 4.5% in children under the age of 3 to 66% in the group of adolescents over the age of 14. The low exposure to HAV infection in younger children from Population 2 could be a result of improved environmental hygiene and sanitation, as demonstrated by the presence of piped water, waste and sewage disposal systems in most houses from this population group. These findings indicate a possible change in the prevalence of hepatitis A in Rio de Janeiro.

Rickettsiae-infected ticks in an endemic area of spotted fever in the State of Minas Gerais, Brazil.

A study on tick-borne rickettsiosis was developed in the county of Santa Cruz do Escalvado, State of Minas Gerais, Brazil, where a clinical case of the disease, confirmed by necropsy, had been reported. Of the 1,254 ticks collected, 1,061 belonged to the Amblyomma genus, 57 to the Rhipicephalus sanguineus species, 81 to Boophilus microplus, and 46 to Anocentor nitens. The hemolymph test associated with Giménez staining showed that 18 of the 221 A. cajennense specimens, 1 of the 16 R. sanguineus, 1 of the 22 B. microplus, 3 of the A. nitens, and 1 of the A. ovale contained rickettsia-like microorganisms. Only 3 A. cajennense ticks were positive under direct immunofluorescence. A. cajennense was the only species found on humans.

Epidemiological aspects of the Brazilian spotted fever: seasonal activity of ticks collected in an endemic area in São Paulo, Brazil

Carrapatos de vegetação e de animais foram coletados mensalmente durante o período de um ano (1993-1994) em uma área endêmica de febre maculosa brasileira no município de Pedreira, São Paulo. Seis espécies de carrapatos foram identificadas Amblyomma cajennense, Amblyomma cooperi, Amblyomma triste, Anocentor nitens, Rhipicephalus sanguineus e Boophilus microplus. Somente a primeira espécie foi suficientemente abundante para permitir um estudo quantitativo com atividade sazonal, embora a distribuição e fonte de captura de outras espécies fossem observadas e aqui relatadas. Estas informações são correlacionadas com a epidemiologia da rickettsiose transmitida por carrapato.

Ticks were collected from vegetation and animals at monthly intervals during one year (1993-1994) in an endemic area of Brazilian spotted fever in the Country of Pedreira, State of São Paulo. Six species of ticks were identified Amblyomma cajennense, Amblyomma cooperi, Amblyomma triste, Anocentor nitens, Rhipicephalus sanguineus and Boophilus microplus. Only the first species was sufficiently numerous to permit a quantitative study with seasonal activity, although the distribution and source of capture of other species were observed and are reported. This information is correlated with the epidemiology of tick-borne rickettsiosis.