Teresinha Tizu Sato Schumaker

Rickettsia infection in five areas of the state of São Paulo, Brazil

This study investigated rickettsial infection in animals, humans, ticks, and fleas collected in five areas of the state of São Paulo. Eight flea species (Adoratopsylla antiquorum antiquorum, Ctenocephalides felis felis, Polygenis atopus, Polygenis rimatus, Polygenis roberti roberti, Polygenis tripus, Rhopalopsyllus lugubris, and Rhopalopsyllus lutzi lutzi), and five tick species (Amblyomma aureolatum, Amblyomma cajennense, Amblyomma dubitatum, Ixodes loricatus, and Rhipicephalus sanguineus) were collected from dogs, cats, and opossums. Rickettsia felis was the only rickettsia found infecting fleas, whereas Rickettsia bellii was the only agent infecting ticks, but no animal or human blood was shown to contain rickettsial DNA. Testing animal and human sera by indirect immunofluorescence assay against four rickettsia antigens (R. rickettsii, R. parkeri, R. felis, and R. bellii), some opossum, dog, horse, and human sera reacted to R. rickettsii with titers at least four-fold higher than to the other three rickettsial antigens. These sera were considered to have a predominant antibody response to R. rickettsii. Using the same criteria, opossum, dog, and horse sera showed predominant antibody response to R. parkeri or a very closely related genotype. Our serological results suggest that both R. rickettsii and R. parkeri infected animals and/or humans in the studied areas.

Ticks (Acari: Ixodidae) from the state of Rondonia, western Amazon, Brazil

Abstract The development, longevity and reproduction of Oligonychus biharensis on four different cultivars of litchi were studied in the laboratory. The total mortality rates from egg to adult on Feizixiao, Baitangying, Ziniangxi and Sanyuehong were respectively 37.27%, 32.45%, 25.52% and 17.32%. The developmental periods from egg to adult varied from 16.97 days on Sanyuehong to 21.05 days on Feizixiao. The average longevity of adult females ranged from 19.72 days on Sanyuehong to 27.01 days on Baitangying, while the oviposition of O. biharensis varied from 68.80 eggs on Baitnagying to 34.00 eggs on Sanyuehong. The daily ovipositon rate of O. biharensis varied from 5.29 eggs on Baitnagying to 1.76 eggs on Feizixiao. The net reproductive rate of increase (R0), intrinsic rates of natural increase (rm) and finite rate of increase (λ) for O. bicharensis on Baitangying were the highest; the three life table parameters (R0, rm and λ) of Oligonychus biharensis on Feizixiao were the lowest. According to the parameters mentioned above, Baitangying litchi was the most suitable host plant and Feizixiao was the most unsuitable host plant for O. biharensis in this study.

Diagnoses of fipronil resistance in Brazilian cattle ticks (Rhipicephalus (Boophilus) microplus) using in vitro larval bioassays.

Fipronil is a phenylpyrazolic insecticide that is widely used in agriculture and has been recently used to control the cattle tick, Rhipicephalus (Boophilus) microplus. Because of the serious problems associated with resistance to the available acaricides, this product has been used as an important alternative to control acaricide-resistant ticks. The objective of this work was to analyse the fipronil sensitivity of ticks that were collected from farms with a history of fipronil use by larval bioassays. A total of 11 Brazilian tick populations were studied: one population from Rio Grande do Sul, one population from Mato Grosso do Sul and nine populations from São Paulo. To validate the assays, susceptible reference strains, POA (Porto Alegre, Brazil) and Mozo (Dilave, Uruguay), and ticks from six different farms that never used fipronil were tested. The resistance of various tick populations to technical grade fipronil (95.3%) was primarily evaluated using the larvae immersion test (LIT) and the larval packet test (LPT), when a sufficient number of larvae was collected. Using the LIT, the resistance ratios (RR(50)) of the tick populations from Rio Grande do Sul and Mato Grosso do Sul were 14.9 and 2.6, respectively, and the populations derived from São Paulo had RR(50)s ranging from 2.5 to 6.9. Four populations were evaluated with the LPT, and two populations displayed lower RR(50), while other populations displayed higher RR(50) than those determined by the LIT. This article reports the first cases of fipronil resistance in Brazil and highlights the LIT as a more sensitive technique for the evaluation of fipronil resistance in R. (B) microplus ticks. We suggest the use of the LIT as an evaluation tool for monitoring fipronil resistance in the control programmes of R. (B) microplus.

Isolation of Rickettsia felis in the mosquito cell line C6/36.

ABSTRACT We report the isolation and establishment of Rickettsia felis in the C6/36 cell line. Rickettsial growth was intense, always with 90 to 100% of cells being infected after few weeks. The rickettsial isolate was confirmed by testing infected cells by PCR and sequencing fragments of three major Rickettsia genes (gltA, ompB, and the 17-kDa protein gene).

Natural Infection, Transovarial Transmission, and Transstadial Survival of Rickettsia bellii in the Tick Ixodes loricatus (Acari: Ixodidae) from Brazil

Abstract:  An Ixodes loricatus engorged female, infected with Rickettsia bellii, was collected from an opossum (Didelphis aurita) in Mogi das Cruzes, São Paulo State, Brazil. Two consecutive laboratory tick generations (F1 and F2) reared from this single engorged female were evaluated for Rickettsia infection by polymerase chain reaction (PCR) targeting specific Rickettsia genes. Immature ticks fed on naïve Wistar rats (Rattus norvegicus) and adult ticks fed on opossum (D. aurita), both free of ticks and rickettsial infection. PCR performed on individual ticks from the F1 (20 larvae, 10 nymphs, and 10 adults) and the F2 (30 larvae, 30 nymphs, and 15 adults) yielded expected bands compatible with Rickettsia. All the PCR products that were sequenced, targeting gltA gene, resulted in sequences identical to each other and 99.7% (349/350) similar to the corresponding sequence of R. bellii in GenBank. The R. bellii infection on ticks from the second laboratory generation (F2) was confirmed by other PCR protocols and successful isolation of R. bellii in cell culture. We report for the first time a Rickettsia species infecting I. loricatus, and the first report of R. bellii in the tick genus Ixodes. We conclude that there was an efficient transovarial transmission and transstadial survival of this Rickettsia species in the tick I. loricatus. Our results suggest that R. bellii might be maintained in nature solely by transovarial transmission and transstadial survival in ticks (no amplifier vertebrate host is needed), since there has been no direct or indirect evidence of infection of vertebrate hosts by R. bellii.

Applicability of in vitro bioassays for the diagnosis of ivermectin resistance in Rhipicephalus microplus (Acari: Ixodidae).

The applicability of laboratory bioassays to diagnose ivermectin (IVM) resistance in Rhipicephalus microplus was evaluated. Adult immersion tests (AITs), larval immersion tests (LITs) and larval packet tests (LPTs) were performed to characterise the effects of ivermectin toxicity on adults and larvae of a susceptible reference strain. The AIT was determined to be a reasonable assay but requires a large number of individuals to attain interpretable results. The LIT and LPT were validated with an IVM resistant strain, revealing resistance ratios (RRs) of 6.73 and 1.49, respectively. In a field survey, nine different populations of cattle tick from the states of São Paulo and Mato Grosso do Sul, Brazil, were analysed with the LIT. Populations without previous exposure to ivermectin exhibited RRs between 0.87 and 1.01. Populations previously exposed to IVM showed RRs between 1.83 and 4.62. The LIT was more effective at discriminating between resistant and susceptible populations than the LPT. The use of the LIT is recommended for the diagnosis of ivermectin resistance in R. microplus.

Rickettsia rickettsii, Rickettsia felis and Rickettsia sp. TwKM03 infecting Rhipicephalus sanguineus and Ctenocephalides felis collected from dogs in a Brazilian spotted fever focus in the State of Rio De Janeiro/Brazil

Brazilian spotted fever (BSF) caused by Rickettsia rickettsii is transmitted to humans by Amblyomma tick species occurring mainly in southeastern regions of the country. The disease is similar to the North American Rocky Mountain spotted fever and without an adequate and prompt treatment it can be fatal. In the last years, the number of BSF cases has increased in the State of Rio de Janeiro. Epidemiologic data on vectors and hosts are lacking. Recently, the first BSF fatal case was registered in the Barra Mansa municipality. Targeting the adoption of the adequate rickettsiosis control strategy we developed an epidemiological study, using immunological and molecular techniques, to detect Rickettsia species in potential vectors and hosts of this new BSF focus.

Prevalence of Rickettsia felis in the Fleas Ctenocephalides felis felis and Ctenocephalides canis from Two Indian Villages in São Paulo Municipality, Brazil

Abstract:  We evaluated the presence of Rickettsia infection among fleas collected on domestic dogs in two Guarani Indian communities in the suburban area of São Paulo Municipality, Brazil. A total of 114 Ctenocephalides felis felis and 47 Ctenocephalides canis were collected from 40 dogs. A total of 41 C. felis felis (36.0%) and 9 C. canis (19.1%) fleas yielded expected bands by PCR, which were all shown by DNA sequencing to be indentical to the corresponding sequence of a fragment of the Rickettsia felis gltA gene deposited in GenBank. The overall prevalence of R. felis was 31.0% (49/161).

Taxonomic and Phylogenetic Relationships Between Neotropical Species of Ticks from Genus Amblyomma (Acari: Ixodidae) Inferred from Second Internal Transcribed Spacer Sequences of rDNA

Abstract The accurate specific identification of ticks is essential for the study, control and prevention of tick-borne diseases. Herein, we determined ribosomal nucleotide sequences of the second internal transcribed spacer (ITS2) of 15 Neotropical hard tick species of the genus Amblyomma Koch found in Brazil. Most of the studied ticks accidentally parasite humans and potentially act as vectors of zoonoses. Lengths of the ITS2 sequences ranged from 956 to 1,207 bp, whereas GC content varied from 62.4 to 66.9%. A matrix of ITS2 divergence was calculated with the ITS2 sequence data obtained showing divergence levels varying from 1.5 to 28.8%. The analysis indicated that this molecular marker can be useful for Amblyomma-specific identification. Phylogenetic inferences based on the ITS2 sequences were used to assess some issues in subgenus taxonomy.

Detecção de riquétsias em carrapatos do gênero Amblyomma (Acari: Ixodidae) coletados em parque urbano do município de Campinas, SP

The city of Campinas is located in an endemic area for brazilian spotted fever in Sao Paulo State, where several cases have recently occurred. Capybaras have been associated with the cycle of this disease, for they present positive serology and serve as host for ticks of the genus Amblyomma, the main vectors of brazilian spotted fever. Ticks were colleted both from Capybaras and from the vegetation in the city park Lago do Cafe, located in the urban area of Campinas city, SP, a site associated with suspected human cases of brazilian spotted fever. The ticks collected were examinaded for the presence of rickettsiae using polymerase chain reaction and the haemolymph test. Through analysis of the gene gltA nucleotide sequence, adults of Amblyomma cajennense and Amblyomma cooperi were found to be infected with the non pathogenic Rickettsia bellii. However, no rickettsiae of the spotted fever group were detected. These results indicate that the role of capybaras as reservoirs of rickettsiae of the Spotted Fever group is still uncertain and further studies are required.