MOLECULAR INVESTIGATION OF HEMOTROPIC MYCOPLASMAS IN HUMAN BEINGS, DOGS AND HORSES IN A RURAL SETTLEMENT IN SOUTHERN BRAZIL

SUMMARY The aims of this study were to determine the prevalence of hemoplasmas in a rural Brazilian settlement's population of human beings, their dogs and horses, highly exposed to tick bites; to identify the tick species parasitizing dogs and horses, and analyze factors associated with their infection. Blood samples from 132 dogs, 16 horses and 100 humans were screened using a pan-hemoplasma SYBR green real-time PCR assay followed by a species-specific TaqMan real-time PCR. A total of 59/132 (44.7%) dog samples were positive for hemoplasmas (21 Mycoplasma haemocanisalone, 12 ' Candidatus Mycoplasma haematoparvum' alone and 21 both). Only 1/100 (1.0%) human sample was positive by qPCR SYBR green, with no successful amplification of 16S rRNA or 23 rRNA genes despite multiple attempts. All horse samples were negative. Dogs >1 year of age were more likely to be positive for hemoplasmas ( p= 0.0014). In conclusion, although canine hemoplasma infection was highly prevalent, cross-species hemoplasma transmission was not observed, and therefore may not frequently occur despite overexposure of agents and vectors.

Rhipicephalus sanguineus (ACARI: IXODIDAE) BITING A HUMAN BEING IN PORTOALEGRE CITY, RIO GRANDE DO SUL, BRAZIL

We report the finding of a female brown dog tick, Rhipicephalus sanguineus (Acari: Ixodidae) on the scalp of a male patient inPorto Alegre, Rio Grande do Sul, Brazil. Human parasitism by this tick is rare and has seldomly been reported in the literature, despite its recognized importance since it can act as a vector of Rickettsia rickettsii, the agent of spotted fever.

Isolamento de Rickettsia em cultura de células vero

Embora o diagnóstico da febre maculosa baseie-se em sinais e sintomas característicos, o mesmo requer confirmação laboratorial, pois existem alguns diagnósticos diferenciais possíveis como meningococcemia, leptospirose, infecção por enterovírus e febre tifóide. A confirmação laboratorial pode ser feita através da pesquisa de anticorpos específicos, possível somente alguns dias após o aparecimento da doença, através do isolamento do agente em amostras de sangue e/ou biópsia de pele, e ainda, de amostras de carrapatos coletados do paciente ou de animais reservatório. O isolamento a partir de sangue ou biópsia de pele resulta em diagnóstico precoce da doença, pois na fase de rickettsemia ainda não há anticorpos detectáveis no sangue. Assim, com o objetivo de facilitar o diagnóstico precoce da febre maculosa, estabelecemos um método de isolamento de rickettsia em cultura de células vero. Para a padronização foi inoculada amostra padrão de Rickettsia rickettsii, cepa Sheyla Smith, cedida pelo CDC. A identificação foi feita através da reação de imunofluorescência indireta. A presença de microrganismos verdes fluorescentes visualizados no interior do citoplasma das células caracterizou o crescimento do agente. Posteriormente, a metodologia foi confirmada pelo isolamento do agente da febre maculosa em amostras de biópsia de pele de paciente proveniente de área endêmica no Estado de São Paulo, bem como, de amostras de carrapato do gênero Amblyomma, considerado o reservatório e transmissor da doença no Brasil.

Analysis of the use of fenthion via epicutaneous in dogs for Rhipicephalus sanguineus control

The action of fenthion was studied in a 15% epicutaneous formulation upon Rhipicephalus sanguineus, which may transmit pathogens to men and other animals, such as Ehrlichia, Babesia and Ricketsia. Dogs were artificially infected for the trial. The fenthion bioassays were begun four months after artificial infestation. The test group, having a mean of 186 ticks per dog, received the formulation dosage according to body weight on the neck region. Tick counts were performed, considering diameters > or = 2mm, during 11 days of treatment, in the most affected body areas: back, ears and paws. Before the application of fenthion in the dogs, it were observed an average 43.3% ticks in the ears, 38.1% in the back area and 17.6% in the paws. The number of ticks in dogs decreased by 36.2%, 63.8%, 82.7%, 67%, 40% and 4.9%, respectively on days 1, 2, 3, 5, 7, 9 and 11 after treatment. R. sanguineus anti-tick activity, lower than that officially recommended, was verified. The number of ticks increased progressively after the 5th day, demonstrating residual insecticide inefficacy. The results obtained did not indicate the use of this formulation, at the tested dosage, as an elective measure for R. sanguineus control.

Study of infection by Rickettsiae of the spotted fever group in humans and ticks in an urban park located in the City of Londrina, State of Paraná, Brazil

INTRODUCTION: Spotted fevers are emerging zoonoses caused by Rickettsia species in the spotted fever group (SFG). Rickettsia rickettsii is the main etiologic agent of Brazilian spotted fever (BSF) and it is transmitted by Amblyomma spp. ticks. METHODS: The study aimed to investigate SFG rickettsiae in the Arthur Thomas Municipal Park in Londrina, PR, by collecting free-living ticks and ticks from capybaras and blood samples from personnel working in these areas. Samples from A. dubitatum and A. cajennense were submitted for PCR in pools to analyze the Rickettsia spp. gltA (citrate synthase gene). RESULTS: All the pools analyzed were negative. Human sera were tested by indirect immunofluorescence assay with R. rickettsii and R. parkeri as antigens. Among the 34 sera analyzed, seven (20.6%) were reactive for R. rickettsii: four of these had endpoint titers equal to 64, 2 titers were 128 and 1 titer was 256. None of the samples were reactive for R. parkeri. An epidemiological questionnaire was applied to the park staff, but no statistically significant associations were identified. CONCLUSIONS: The serological studies suggest the presence of Rickettsiae related to SFG that could be infecting the human population studied; however, analysis of the ticks collected was unable to determine which species may be involved in transmission to humans.

Risk factors associated with the transmissionof Brazilian spotted fever in the Piracicaba river basin, State of São Paulo, Brazil

INTRODUCTION : Brazilian spotted fever (BSF) is a disease transmitted by ticks for which the etiological agent is Rickettsia rickettsii. The present essay evaluates the risk factors associated with the transmission of cases of BSF in the time period between 2003 and 2013 in the Piracicaba river basin, state of São Paulo. METHODS : This essay presents a retrospective study to identify the factors associated with the transmission of cases of BSF among all suspected cases identified by the System for Epidemiological Surveillance of São Paulo (CVE). After the description of temporal distribution (onset of symptoms) and the environmental and demographic variations of the confirmed and discarded cases, a multiple logistic regression model was applied. RESULTS : We searched 569 probable locations of infection (PLI) with 210 (37%) confirmed cases of BSF and 359 (63%) discarded cases. The associated variables for the confirmation of BSF in the multiple logistic model using a confidence interval (CI) of 95% were age (OR = 1.025 CI: 1.015-1.035), the presence of Amblyomma sculptum in the environment (OR = 1.629 CI: 1.097-2.439), the collection of ticks from horses (OR = 1.939 CI: 0.999-3.764), the presence of capybaras (OR = 1.467 CI: 1.009-2.138), an urban environment (OR = 1.515 CI: 1.036-2.231), and the existence of a dirty pasture (OR = 1.759 CI: 1.028-3.003). CONCLUSIONS : The factors associated with the confirmation of BSF cases included an urban environment, age, presence of the A. sculptum vector, the collection of ticks from horses, the presence of a capybara population, and a dirty pasture environment.

Ectoparasitos de pequenos mamíferos da Ilha de Maracá, Roraima, Brasil. I. Ectoparasitofauna, registros geográficos e de hospedeiros.

Um levantamento de ectoparasitos de pequenos mamiferos foi realizado na ilha de Ma-racá, Roraima, Bnasil, durante três distintos periodos entre novembro de 1987 a fevereiro de 1989, tendo sido colecionados 1. 774 exemplares retirados de 51 roedores e 3 marsupiais. Os números e a distribuição por sexo das espécies de ectoparasitos encontrados são apresentados. Novos registros de hospedeiros são dados pana Androlaelaps fahrenhol zi, Gigantolaelaps goyanensis, Laelaps flexa, Laelaps paulistanensise Tur aymara.Todas as espécies de ectoparasiZos são registradas pela primeira vez no Estado de Rorai ma. Excetuando-se Amblyomma.sp., Cummingsiasp., Hoplopleura splendida, Polygenis kla-gesi klagesi, Polygenis klagesi samuelise Rhopalopsyllus australisssp. as demais espé-cies são assinaladas pela primeira vez na Amazônia brasileira. L. flexa, Tur amazonicus, Turapicalis, Τ. aymanae Gliricola venezuelanustambém constituem novos negistros pora o Bnasil. Duas novas espécies de. ectoparasitos - uma de ácaro, outra de malófogo-foram, também, encontradas.

The parasitism of Ixodes luciae (Acari: Ixodidae) on marsupials and rodents in Peruvian Amazon

In this work the infestation with I. luciae on Didelphimorphia and Rodentia in different environments of Peruvian Amazon was studied. Didelphimorphia was represented by the family Didelphidae. Specimens belonging to Caluromys lanatus, Didelphis marsupialis, Marmosops sp.2, Metachirus nudicaudatus, Philander andersoni and Philander opossum were infested with adults I. luciae and one Micoureus sp. was infested with larvae. In Rodentia, the infestation with I. luciae nymphs was restricted to Hylaeamys perenensis, Hylaeamys yunganus and Oligoryzomys microtis, while one Oecomys bicolor (all Cricetidae) was infested with larvae of this species. The few larvae were found on rodents captured in primary forest. The only significant difference (P < 0.05) in prevalence of adult ticks on Didelphimorphia was between P. andersoni and M. nudicaudatus (chi-square distribution). Adult tick distribution was significant different in P. andersoni in comparison with M. nudicaudatus, P. opossum and D. marsupialis (Kruskal-Wallis test). No significant effect of month or environment was detected in relation to adult tick infestation on Didelphimorphia. The prevalence of nymphal infestation as well as tick distribution showed that H. perenensis and H. yunganus were significantly more prone to be infested with nymphs of I. luciae than O. microtis. Prevalence of nymph infestation was higher in primary and secondary forest than rural areas while abundance was higher in secondary forest when compared with rural areas (P < 0.05). Kruskal-Wallis test showed differences (P < 0.05) for nymphal infestation during December in relation to January, March, April and June. The natural cycle of I. luciae appeared to be continuous, bound to adult tick infestation on Philander and nymphal infestation on Hylaeamys in forested environs.

Observações sobre os Ixodideos da Republica Argentina

Ixodidae of the Argentine Republic are studied by the Author based on 71 lots of material, sent by Prof. Salvador Mazza, comprising 13 different species, two of which are found in Argentine territory: Haemaphysalis kochi Arag. and Amblyomma parvitarsum Nn. The author admits the existence in Argentina of 23 species of Ixodidae, but such number may still increase along with the development of the studies on the subject. In this paper, the Ornithodoros classified by Barbará and Dios as Ornithodoros turicata, is sunk by the Author in synonymy with O. rostratus Arag. not merely on the strength of the material at his disposal, but also by the plates published by the Argentine authors. The following species are also placed in synonymy: Amblyomma altiplanum Dios, 1917 (= Amblyomma parvitarsum Nn., 1899), Amblyomma concolor, Nn., 1899 (=Amblyomma auriculare Conil, 1877) Amblyomma furcula Donitz, 1907 (= Amblyomma neumani Ribaga, 1902). In the Author's opinion Amblyomma striatum C. L. Koch, 1844 and Amblyomma fossum Nn. are distinct species, so that the same Amblyomma ovale for both should no longer subsist as L. C. Robinson proposes. The Author, moreover, shows his doubts upon the existence in Argentine of Aponomma laeve, Nn., 1899 and Dermacentor triangulus Nn., 1899, as they have not been seen any more and in South-America no representatives of the genera Aponomma and Dermacentor are known.

Nota sobre os Ixodideos da Republica Argentina

A study of and addicional collection of Argentine ixodidae is made on material send to the Author by Professor Salvador Mazza of Departamento de pathologia do Norte - Jujuy and Professor Doello Jurado of the Museu Rivadavia. There was examined 37 lots of Ixodidae. Among them there were found the nymph of Ixodes auritulus Nn. and 3 other species already known on the South American fauna and from which there was til yet no record in Argentine; they are Ixodes ricinus, Amblyomma parvum and Amblyomma pseudo-concolor.

Verificação, em condiçoes naturais, da infecção de mais três hematófagos (Anophelineos, Flebótomos e Simulídeos) em leprosos

The A. went last August to the State of Minas Gerais to continue his studies on transmission of leprosy by insects. He selected ten lepers (all L3 cases) for his experiments. It happened in the middle of August, a few day after freezing temperature. Practically there were no day mosquitoes, even near a river where in March there were very many. Bringing the patients to a wood, near the Peixe River (Fish River), at the dawn they were attacked by very many Anopheles, some flebotomus, a few Simulium and very rare Culex scapularis. All these insects became infected, in different degrees, by biting such patients. CONCLUSIONS. 1. Two species of Anopheles (A. albitarsis and A. tarsimaculatus) became strongly infected by Hansen bacilli. 2. By dissection done by Dr. Oliveira Castro were found lepra bacilli in various points of the proboscis of two Flebotomus (F. intermedius) and in their stomachs. 3. By smearing were found lepra bacilli in two specimens of Simulium sp. (probably pertinax). 4. It was confirmed also the verification done last March, at the same Leper Colony, that Phthirius pubis can be also a carrier of lepra bacillus. 5. There was confirmed also natural infection of nymphs of Amblyomma cajennense in lepers. Dr. Oliveira Castro is dissecting the Anopheles to locate the bacilli in their organisms and he started, with the cooperation of the Director of Colonia Santa Fé, Dr. José Mariano, attenpts to re-infect a group of negative-nerve cases of leprosy with infected mosquitoes.

Culturas de bacilos ácido-álcool resistentes isolados de hematófagos infectados em leprosos: evidências de se tratar do bacilo de Hansen

The A. summarises the history of his first culture of acidfast bacillus isolated directly from leprosy lesions (Sample José) and refers about two samples recovered from guinea pig and white rat inoculated with said culture. Then the A. completes his previous descriptions of four cultures of acidfast bacilli isolated by him from ticks (Amblyomma cajaennense and Boophilus microplus, two cultures from each species) infected experimentally in lepers. The A. having found specimens of two species of Triatomidae (Triatoma infestans and Panstrongylus megistus) naturally infected with HANSEN bacillus in huts habited by lepers in the State of Minas Gerais (Dec. 1942), started a series of experiments, using larvae and nymphs of T. infestans bred in laboratory at the Instituto Oswaldo Cruz, to infect in active cases of leprosy, in the city of Rio de Janeiro, could obtain two new samples of cultures of acid-fast bacilli (Ns. 6 and 7 of his set). In this papaer the A. studies the biological properties of said cultures, proving that Penicilin has not effect upon them, like other substances. The sulphuric and acetic acids were used to purify some of the cultures, with good results, the cultures becoming more rich and growing faster. Potassium hydroxide Sodium (10% solution) was also used with success to isolate and to purify the cultures, but it seems that it affects the bacilli in some way. In flud glycerinated media the majority of such cultures produce velum suitable for the preparation of antigens for skin tests and for therapeutical use. At last the A. says that he is becoming convinced that the HANSEN bacillus is in cause, especially after thee evidences of culturing the bacillus from one patient, in different opportunities.

Profilaxia do tifo exantemático neotrópico no Brasil

A profilaxia racional de um doença decorre do melhor e mais profundo conhecimento dela. A do tifo exantematico neotrópico, apesar do que se sabe da doença, não e fácil no Brasil. Uma das maiores dificuldade encontramos nas distancias do nosso "hinterland" e na falta de cultura dos nossos agricultores. O homem se infecta certamente com as raças VB, VA e VA do virus brasileiro: 1.°) no campo, nas matas e nas macegas; 2.°) dentro dos domicílios ou nos arredores dêstes. Os primeiros constitúem de regra, os casos insulados do mal. Os segundos fazem parte, não raro, dos focos domiciliares macicos, com 2, 5, 7 e até 12 casos na mesma casa. São responsáveis, pelos primeiros, os carrapatos, principalmente o Amblyomma cayennense e o Amblyomma brasiliense, mormente quando no estado de ninfas, dada a herança habitual das infecções nesses artrópodos, que se infestam em animais silvestres, depositários do virus. São responsáveis pelos segundos, os "Cimex lectularius", percevejos dos leitos e as ninfas e larvas dos carrapatos, deixadas cair junto aos domicílios ou mesmo dentro deles. A profilaxia racional e completa da doença entre nós compreende: 1.°) Descarrapatização das zonas infestadas, por meio de leis apropriadas, coercitivas e aplicadas sem excepção; 2.°) Combate aos cães vadios, cabritos e outros animais portadores do virus; 3.°) Queima dos pastes, campos e macegas de fraco valor econômico, principalmente os que confinam com residências; 4.°) A propaganda racional contra esta grave doença exantemática: a) com palestras locais, acompanhadas de fotografias e gráficos expressivos e ao alcance de todos; b) pelo cinema; c) com artigos simples, claros e precisos sôbre a matéria; d) com folhetos apropriados; 5.°) Combater toda a vermina dentro e nas proximidades dos domicílios, 6.°) Demonstrar que é possível, com toda a certeza, evitar a doença, retirando os carrapatos que se prenderem ao corpo, dentro de 12 ou 14 horas após a fixação; 7.°) Aconselhar o emprego do DDT, “Gammexame" e “Toxafeno" para o expurgo das casas e animais, a fim de combater os carrapatos; 8.°) Vacinação preventiva contra a doença, quando possível, em zonas delimitadas e já civilizadas, feitas principalmente com raças de virus colhidas no Brasil e, se possível, com uma só dose. O autor trabalhou durante algum tempo com a vacina tipo Spencer-Parker. Hoje aconselha o trabalho com a vacina tipo Cox, original ou modificada.

Interrelação entre acari Ixodidae e hospedeiros Edentata da Serra da Canastra, Minas Gerais, Brasil

Recebemos para exame uma pequena coleção de carrapatos no Parque Nacional da Serra da Canastra (MG), entre 1979 e 1980. Os autores demosntraram a existência de uma larga co-acomodação de Amblyomma pseudoconcolor em edentados da familia Dasypodidae, sendo Dasypodini a tribo mais ajustada a sta infestação. De acordo com as Figs. 1 e 2, Dasypodini são, provavelmente, os hospedeiros reais de A. pseudoconcolor e também os hospedeiros mais antigos. Pela primeira vez, A. pseudoconcolor é também registrado e, Cabassous tatouay, C. unicinctus, priododntes maximus e Euphractus sexcinctus. Também, pela primeira vez, A. pseudoconcolor e A. calcaratum foram registrados no Estado de Minas Gerais. os ectoparasitos estão depositados no Departamento de Parasitologia da Universidade Federal de Minas Gerais, Brasil.

Ectoparasites Ixodida Leach, 1817 on wild mammals in the state of Paraná, Brazil

A taxonomical and ecological study was made on 264 samples of 12 species of ectoparasite ixodides collected on wild mammals from several natural regions of Paraná state, Brazil. These species of ticks as their hosts are listed by their identified evolutive forms and capture locations. A review of the early studies on the Ixodida from South Brazil is made considering the frequency of the parasite species on each host concerning the geographic distribution of such species. This paper is a contribution to the knowledge of the Ixodidae ectoparasites species of wild mammals in Brazil.