Wide Dispersal and Possible Multiple Origins of Low-Copy-Number Plasmids in Rickettsia Species Associated with Blood-Feeding Arthropods

Plasmids are mobile genetic elements of bacteria that can impart important adaptive traits, such as increased virulence or antibiotic resistance. We report the existence of plasmids in Rickettsia (Rickettsiales; Rickettsiaceae) species, including Rickettsia akari, ""Candidatus Rickettsia amblyommii,"" R. bellii, R. rhipicephali, and REIS, the rickettsial endosymbiont of Ixodes scapularis. All of the rickettsiae were isolated from humans or North and South American ticks. R. parkeri isolates from both continents did not possess plasmids. We have now demonstrated plasmids in nearly all Rickettsia species that we have surveyed from three continents, which represent three of the four major proposed phylogenetic groups associated with blood-feeding arthropods. Gel-based evidence consistent with the existence of multiple plasmids in some species was confirmed by cloning plasmids with very different sequences from each of two ""Ca. Rickettsia amblyommii"" isolates. Phylogenetic analysis of rickettsial ParA plasmid partitioning proteins indicated multiple parA gene origins and plasmid incompatibility groups, consistent with possible multiple plasmid origins. Phylogenetic analysis of potentially host-adaptive rickettsial small heat shock proteins showed that hsp2 genes were plasmid specific and that hsp1 genes, found only on plasmids of ""Ca. Rickettsia amblyommii,"" R. felis, R. monacensis, and R. peacockii, were probably acquired independently of the hsp2 genes. Plasmid copy numbers in seven Rickettsia species ranged from 2.4 to 9.2 per chromosomal equivalent, as determined by real-time quantitative PCR. Plasmids may be of significance in rickettsial evolution and epidemiology by conferring genetic plasticity and host-adaptive traits via horizontal gene transfer that counteracts the reductive genome evolution typical of obligate intracellular bacteria.

Transovarial passage of Leishmania infantum kDNA in artificially infected Rhipicephalus sanguineus

Phlebotomine sand flies are the only proven biological vectors of Leishmania parasites. However, Rhipicephalus sanguineus ticks have long been suspected to transmit Leishmania infantum in studies carried out in laboratory and natural conditions. In the present study, 5 mu l of L. infantum promastigotes (1 x 10(6) cells per ml) was injected into the hemocel through the coxa 1 of four engorged females (F1, F2, F3 and F4). Control ticks (F5 and F6) were injected with sterile phosphate-buffered saline (PBS) using the same procedure. Then, these females, their eggs, and the originated larvae were tested by real time polymerase chain reaction (real-time PCR) for the presence of L. infantum kinetoplast DNA (kDNA). Females and eggs were tested after the end of the oviposition period (about 5 weeks post-inoculation) whereas larvae were tested about 4 months after the inoculation of females. All artificially infected females were positive for L. infantum kDNA. In addition, two pools of eggs (one from F2 and other from F4) and four pools of larvae (one from each F1 and F4 and two from F2) were positive for L infantum kDNA. These results showed, for the first time, the transovarial passage of L. infantum kDNA in R. sanguineus ticks, thus suggesting that the transovarial transmission of L. infantum protozoa in ticks is worth to be investigated. (C) 2010 Elsevier Inc. All rights reserved.

Recurrent gains and losses of large (84-109 bp) repeats in the rDNA internal transcribed spacer 2 (ITS2) of rhipicephaline ticks

We studied the internal transcribed spacer 2 (ITS2) in twenty-two spp. of ticks from the subfamily Rhipicephalinae. A 104-109 base pair (bp) region was Imperfectly repeated In most ticks studied. Mapping the number of repeat copies on to a phylogeny from the ITS2 showed that there have been many Independent gains and losses of repeats. Comparison of the sequences of the repeat copies Indicated that in most taxa concerted evolution had played little if any role in the evolution of these regions, as the copies clustered by sequence position rather than species, In our putative secondary structure, each repeat copy can fold into a distinct and almost identical stem-loop complex; a gain or loss of a repeat copy apparently does not impair the function of the ITS2 in these ticks.

Host plant specificity in several species of generalist mite predators

1. Species in the genus Neoseiulus are considered to be generalist predators. with some species used in biological control programmes against phytophagous mites and insects. 2. A general survey of Neoseiulus species in inland Australia indicated that different species are associated with particular tree species. This pattern of host plant use was investigated for four Neoseiulus species (N. buxeus, N. cappari, N. brigarinus, N. eremitus) by means of a sampling programme through time and across space. 3. Each species of Neoseiulus was collected entirely or mostly from one species of tree: little or no overlap was detected despite the tree species growing in well-mixed stands. Host plant specificity thus appears to be strong in this genus. 4. Species in two other genera (Pholaseius and Australiseiulus), also considered to be predatory, showed a similar association with particular tree species. 5. The implications for the use of these predators in biological control are considerable. In particular, phytoseiid species with specific needs in terms of host plants may not be suitable for use as general purpose predators. Meeting the needs of phytoseiids through the modification of host plant attributes may be a step towards enhancing their efficacy as biological control agents.

Low intraspecific variation in the rRNA internal transcribed spacer 2 (ITS2) of the Australian paralysis tick, Ixodes holocyclus

Ixodes holocyclus has a narrow, discontinuous distribution along the east coast of Australia. We studied ticks from 17 localities throughout the geographic range of this tick. The ITS2 of I. holocyclus is 793 bp long. We found nucleotide variation at eight of the 588 nucleotide positions (1.4%) that were compared for all ticks. There were eight different nucleotide sequences. Most sequences were not restricted to a particular geographic region. However, sequences F, G and H, which had an adenine at position 197, were found only in the far north of Queensland - all other ticks had a guanine at this position. The low level of intraspecific variation in this tick (0.7%) contrasts with the sequence divergence between L holocyclus and its close relative, I. cornuatus (13.1 %). These data indicate that L holocyclus does not contain cryptic species despite possible geographic isolation of some populations. We conclude that variation in the ITS2 is likely to be informative about the phylogeny of the group.

The mitochondrial 12S gene is a suitable marker of populations of Sarcoptes scabiei from wombats, dogs and humans in Australia

We sequenced part of the mitochondrial 12S ribosomal RNA gene of 23 specimens of Sarcoptes scabiei from eight wombats, one dog and three humans. Twelve of the 326 nucleotide positions varied among these mites and there were nine haplotypes (sequences) that differed by 1-8 nucleotides. Phylogenetic analyses indicated that these mites were from two lineages: (1) mites from wombats from Victoria, Australia, and mites from the humans and dog from the Northern Territory, Australia (haplotypes 1-4, 9); and (2) mites from the humans and dog from the Northern Territory (haplotypes 5-8). Mites from the three different hosts (wombats, a dog and humans) had not diverged phylogenetically; rather, these mites had similar 12S sequences. Thus, we conclude that these mites from wombats, humans and a dog are closely related, and that they diverged from a common ancestor relatively recently. This conclusion is consistent with the argument that people and/or their dogs introduced to Australia the S. scabiei mites that infect wombats Australia. So, S. scabiei, which has been blamed for the extinction of populations of wombats in Australia, may be a parasitic mite that was introduced to Australia with people and/or their dogs. These data show that the mitochondrial 12S rRNA gene may be a suitable population marker of S. scabiei from wombats, dogs and humans in Australia.

A new subfamily, Bothriocrotoninae n. subfam., for the genus Bothriocroton Keirans, King & Sharrad, 1994 status amend. (Ixodida : Ixodidae), and the synonymy of Aponomma Neumann, 1899 with Amblyomma Koch, 1844

Evidence suggesting polyphyly of the traditionally recognised tick genus Aponomma Neumann, 1899 is summarized. Continued recognition of this genus in its current concept leaves a polyphyletic genus Aponomma and a paraphyletic genus Amblyomma Koch, 1844. To improve the correlation between our understanding of phylogenetic relationships in metastriate ticks and their classification, a few changes in classification are proposed. The members of the 'indigenous Australian Aponomma' group (sensu Kaufman, 1972), A. auruginans Schulze, 1936, A. concolor Neumann, 1899, A. glebopalma Keirans, King & Sharrad, 1994, A. hydrosauri (Denny, 1843) and A. undatum (Fabricius, 1775), are transferred to Bothriocroton Keirans, King & Sharrad, 1994, which is raised to full generic rank. The remaining members of Aponomma are transferred to Amblyomma. Uncertainty remains on relationships of Bothriocroton to other metastriate lineages and on the systematic position of the two species formerly included in the 'primitive Aponomma' group, A. elaphense Price, 1959 and A. sphenodonti Dumbleton, 1943.

A survey of bacterial diversity in ticks, lice and fleas from Australia

We isolated bacteria from ticks, lice and fleas. Partial small subunit rRNA sequences were obtained for each isolate and the closest matches in the FastA database were determined. These bacteria were mostly Gram-positive (Firmicutes), although representatives from the Proteobacteria (alpha, beta, gamma subdivisions) and CFB group were also isolated. Most of the isolates we found were from genera that were present in most of the ectoparasites studied, but a few genera were restricted to one species of ectoparasite. The most commonly isolated genera were Stenotrophomonas, Staphylococcus, Pseudomonas, Acinetobacter and Bacillus. Species of Bacillus and Proteus, which have biopesticide potential, were found in some of these ectoparasites. Overall, the communities of bacteria were similar to those found in other studies of parasitic arthropods.

A new mite from an arboreal ant (Formicidae : Polyrachis sp.): Myrmozercon iainkayi n. sp (Mesostigmata : Laelapidae)

Previously, two species of Myrmozercon (= Myrmonyssus) have been described from ground-nesting Australian ants in the genus Iridiomyrmex. Herein a new species, Myrmozercon iainkayi, infesting workers and alate adults of a leaf-nesting species of Polyrachis ant is described. The new species has a number of unusual or unique characters, including coxal hypertrichy (6-6-6-4 for legs I-IV, respectively), and based on the structure of its mouthparts, appears to be a haemolymph-feeding parasite.

Identificação de genoespécies do complexo Borrelia burgdorferi sensu lato (s.l.) circulantes na fauna ixodideológica em portugal

Dissertação para obtenção do grau de Mestre em Microbiologia Médica

Presença e alterações da fauna de Artrópodos durante a preparação de um composto orgânico

No decorrer da preparação de um composto orgânico (150 dias de duração), foram feitas cinco amostragens para estudarem-se a presença e as alterações da fauna de artrópodos, durante algumas das diferentes fases de sua preparação. Foram utilizados, em seis canteiros feitos acima da superfície do solo, três tratamentos diferentes, além da inoculação bacteriana de três dos canteiros, aos 75 dias, e da cobertura de todos os canteiros com plástico durante uma boa parte do tempo de preparação do composto. Em todos os tratamentos, o grupo numericamente mais importante foi Acari (88-99, 5%), seguido por Collembola (0,3- 0%), Coleoptera (0, 1-1, 4%), Diplopoda (0,04-0,3%), Diptera (0,04-0,65%) e Formicidae (0,01-0,03%). O número de grupos taxonômicos e de indivíduos da fauna aumentou gradativamente com o tempo de preparação do composto, em todas os tratamentos. A inoculação bacteriana, aos 75 dias, resultou num incremento da fauna no tratamento 2 (com esterco de galinha), provavelmente em face do anterior fornecimento de uma eficiente fonte de nitrogênio e de microorganismos, representada pela adição inicial de esterco de galinha fresca ao material vegetal. Também a cobertura dos canteiros com plástico parece ter influido bastante na fauna presente no material, provocando uma sensível diminuição de sua densidade. O tratamento com esterco de galinha foi o que apresentou uma mais rápida colonização pela fauna, com uma maior diversidade de grupos taxonômicos e também o que teve uma mais rápida e completa humificação do material.

Ectoparasitos de cães e gatos da cidade de Manaus, Amazonas, Brasil

São apresentados resultados da coleta de ectoparasitos em cães e gatos entre agosto de 2001 e maio de 2002 em diferentes bairros da cidade Manaus. No cão foram encontrados: Ctenocephalides f. felis (Bouché, 1835) (Siphonaptera, Pulicidae), Heterodoxus spiniger (Enderlein, 1909)(Phthiraptera, Boopidae), Trichodectes canis (De Geer, 1778) (Phthiraptera, Trichodectidae) e Rhipicephalus sanguineus (Latreille,1806) (Acari, Ixodidae). No gato foi coletado C. f. felis. A prevalência de ectoparasitos foi de 80,8% para cães e 72,7% para gatos. Para a pulga C. f. felis foi de 28,7% para cães e 72,7% para gatos. Para o piolho H. spiniger foi de 12,3% para cães. Para o piolho T. canis foi de 0,1% para cães e para o carrapato R. sanguineus foi de 63% para cães. A média de infestaçãode pulga foi de 1,26 para cães e 1,27 para gatos. A proporção sexual fêmea/macho foi de 1,96:1 no cão e de 3,66:1 no gato. A pulga C. canis (Curtis, 1826), registrada em 1922, não foi coletada.

Ácaros encontrados sôbre maçãs de procedência argentina

Apples from Argentina showed large numbers of mite eggs and some diapausing females. They were reared on leaves of lima beans. From the eggs acari related to Panonychus ulmi were obtained and also specimens of Cheyletidae. The diapausing females proved to be Tetranychus telarius and large populations were obtained from four females. There was also one specimem of Phytoseiid mite.

Dois ácaros associados à abelha (Apis mellifera L.) no Perú

Larvas de Leptus sp. (Acari, Prostigmata, Erythraeidae) foram coletadas das membranas intersegmentares dos tergitos e esternitos abdominais de abelhas (Ápis mellifera L.) em Cerro de Pasco, Peru. Não foram relatados danos por este ectoparasito. Fêmeas de Blattisoeius dentriticus (Berlese, 1918) (Acari, Mesostigmata, Ascidae) foram coletadas de colméias de abelhas, junto com Tyrophagus putrescentiae (Schrank, 1781) (Acari, Astigmata, Acaridae), em Lima, Peru.

Ácaros do pinheiro-do-Paraná em Lages, Santa Catarina

Coletas quinzenais de ácaros de ramos do Pinheiro-do-Paraná, Araucária angustifolia, em Lages, Santa Catarina, revelaram a presença, durante um ano (Setembro de 1983 a Agosto de 1984) das espécies fitófagas Oligonychus urunguis (Jacobi) e Tydeus formosus (Cooreman) e de ácaros predadores da família Phytoseiidae, dos gêneros Amblyseius e Proprioseiopsis. Também ácaros oribatídeos foram coletados durante todo o ano. De ocorrência mais restrita foram obtidos ácaros das famílias Ascidae, Bdellidae, Cheyletidae e Acaridae. O método de coleta batendo-se o material vegetal sobre tela e bandeja revelou-se pouco mais eficiente do que o emprego do funil de Tullgren.