Phylogenetic insights on Mediterranean and Afrotropical <Emphasis Type="Italic">Rhipicephalus</Emphasis> species (Acari: Ixodida) based on mitochondrial DNA

This study was performed to determine the tick species that infest cattle and humans throughout an altitudinal gradient in the Yungas Biogeographic Province of Argentina. The presence of tick-borne bacteria of the genera Rickettsia, Ehrlichia and Borrelia in the collected ticks was also evaluated. Samples of ticks parasitizing cattle and humans were carried out in different seasons. Questing ticks (adults and nymphs) were collected from vegetation and analyzed to detect the presence of Rickettsia, Ehrlichia and Borrelia by a battery of different PCRs. Five species of hard ticks were found parasitizing cattle: Amblyomma sculptum, Amblyomma tonelliae, Amblyomma hadanii, Haemaphysalis juxtakochi and Ixodes pararicinus. Amblyomma sculptum (immature and adults), A. tonelliae (immature and adults), A. hadanii (larvae) and one nymph of I. pararicinus were found attached to humans. Rickettsia amblyommatis was detected in one nymph of A. hadanii. DNA of a Borrelia genospecies belonging to the B. burgdorferi s.l. complex (phylogenetically related to haplotypes previously reported in Ixodes aragaoi from Uruguay and I. pararicinus from Argentina) was detected in adults of I. pararicinus. Amblyomma sculptum and I. pararicinus appear to be the tick species more frequent on cattle in the YBP from Argentina, and A. sculptum and A. tonelliae, were the main ticks found attached to humans. The medical importance of the bacteria of the genus Rickettsia and Borrelia detected in this work remains unknown.     

Host range and geographical distribution of <Emphasis Type="Italic">Ixodes sigelos</Emphasis> (Acari: Ixodidae)

Larvae, nymphs and females of Ixodes sigelos Keirans, Clifford and Corwin (Ixodidae) were collected in 13 localities of the Patagonian region of Argentina parasitizing eight species of sigmodontine rodents. We report for the first time adults of I. sigelos for Argentina. Besides, we extend the southern limit of its geographical distribution, and six species of sigmodontines are added as new host species of I. sigelos (Phyllotis xanthopygus, Euneomys chinchilloides, Calomys musculinus, Reithrodon auritus, Loxodontomys micropus and Eligmodontia morgani). The presence of larvae, nymphs and females on sigmodontines, as well as more than 50% of the individuals engorged, indicate that I. sigelos can develop the complete parasitic phase of its life cycle on these small mammals. The geographical distribution of I. sigelos is restricted to the biogeographical Andean Region in Argentina and Chile.     

First record of the avian ectoparasite <Emphasis Type="Italic">Philornis downsi</Emphasis> Dodge &; Aitken, 1968 (Diptera: Muscidae) in Argentina

Species of Philornis Meinert, 1890 (Diptera, Muscidae) are Neotropical dipterans that include species with parasitic larvae which feed on nestling birds. To date, all Philornis species that have been recorded from Argentina have parasitic subcutaneous larvae. Here, for the first time for Argentina, we report the finding of Philornis downsi Dodge & Aitken, 1968, a fly with a nest-dwelling, semi-haematophagous larva. This record, from the humid Chaco ecoregion of Argentina in the nest of a saffron finch Sicalis flaveola pelzelni Sclater, substantially extends the known distribution of this species. We also report the consensus sequences of the internal transcribed spacer 1 (ITS1) and ITS2 regions of three of the specimens for future reference and comparison. Further investigation is needed to determine whether Argentina is part of the historical range of P. downsi or, alternatively, represents a recent expansion of its range, perhaps due to climatic changes or other factors of global environmental variation.     

Host-parasite association between <Emphasis Type="Italic">Didelphis albiventris</Emphasis> (Didelphimorphia: Didelphidae) and <Emphasis Type="Italic">Ixodes loricatus</Emphasis> (Acari: Ixodidae) in their southern ranges

The parasitism of Ixodes loricatus Neumann on white-eared opposum, Didelphis albiventris Lund, was analysed in the southern ranges of both. In central Argentina, 118 wild opossums were captured from 2005 to 2012. Adults of I. loricatus were collected (prevalence?=?66.1%, mean?=?7.03, median?=?4), but no immature stage. Tick infestation was not affected by sex (p?=?0.27) or age (p?=?0.37) of hosts. In line with previous studies about seasonality of larvae and nymphs of I. loricatus, adult tick infestation showed no seasonal trend. This lack of seasonal pattern of infestation for all parasitic stages of I. loricatus may indicate a life cycle governed by nidicolous behaviour, in such a way that this habit would contribute to minimize the influence of external environmental variables. The tick sex ratio was female biased, with two females per one male (p?=?0.002). The absence of I. loricatus immatures on D. albiventris suggests that in the study region there is clear segregation of immatures in rodents and adults in marsupials.     

Differential diagnosis of <Emphasis Type="Italic">Ixodes ricinus</Emphasis> and <Emphasis Type="Italic">Ixodes persulcatus</Emphasis>: nymphs and larvae

We developed a method for differential diagnosis of nymphs and larvae of sheep (Ixodes ricinus (L.)) and taiga (I. persulcatus Sch.) ticks (Parasitiformes: Ixodidae) which allows to identify live material in the field.     

Pathogenicity of <Emphasis Type="Italic">Metarhizium anisopliae</Emphasis> (Deuteromycetes) and permethrin to <Emphasis Type="Italic">Ixodes scapularis</Emphasis> (Acari: Ixodidae) nymphs

Effectiveness of the entomopathogenic fungus Metarhizium anisopliae, for controlling nymphal Ixodes scapularis, was tested in laboratory and field trials. In the laboratory, M. anisopliae (Metschnikoff) Sorokin strain ESC1 was moderately pathogenic, with an LC₅₀ of 10⁷ spores/ml and induced 70% mortality at 10⁹ spores/ml. In a field study, however, 10⁹ spores/ml M. anisopliae did not effectively control questing I. scapularis nymphs, and significant differences were not detected in pre- and post-treatment densities. For nymphs collected and returned to the laboratory for observation, mortality was low in treatment groups, ranging from 20 to 36%. To assess whether a chemical acaricide would synergistically enhance pathogenicity of the fungus, we challenged unfed nymphal I. scapularis with combinations of M. anisopliae and permethrin, a relatively safe pyrethroid acaricide, in two separate bioassays. Significant interactions between M. anisopliae and permethrin were not observed, supporting neither synergism nor antagonism.     

Repellency of two terpenoid compounds isolated from <Emphasis Type="Italic">Callicarpa americana</Emphasis> (Lamiaceae) against <Emphasis Type="Italic">Ixodes scapularis</Emphasis> and <Emphasis Type="Italic">Amblyomma americanum</Emphasis> ticks

Callicarpenal (13, 14, 15, 16-tetranor-3-cleroden-12-al) and intermedeol [(4S,5S,7R,10S)-eudesm-11-en-4-ol], isolated from American beautyberry, Callicarpa americana (Lamiaceae), were evaluated in laboratory bioassays for repellent activity against host-seeking nymphs of the blacklegged tick, Ixodes scapularis, and lone star tick, Amblyomma americanum. A strip of organdy cloth treated with test solution was doubly wrapped (treatment on outer layer) around the middle phalanx of a forefinger and ticks released on the fingertip. Callicarpenal and intermedeol, at 155 nmole/cm² cloth repelled 98 and 96% of I. scapularis nymphs, respectively. Dose response tests with I. scapularis nymphs showed no difference in repellency among callicarpenal, intermedeol and Deet (N,N-diethyl-3-methylbenzamide), however, SS220 ((1S,2′S)-2-methylpiperidinyl-3-cyclohexene-1-carboxamide) was significantly more repellent than the other compounds. Callicarpenal, at 155 nmole/cm² cloth, repelled 100 and 53.3% of I. scapularis nymphs at 3 and 4 h, respectively, after the cloth was treated, whereas intermedeol repelled 72.5% of I. scapularis nymphs 3 h after treatment. In comparison with the results obtained with I. scapularis, callicarpenal, intermedeol, Deet and SS220 were less effective against A. americanum. Only intermedeol and SS220 repelled significantly more A. americanum than ethanol controls at 155 nmole compound/cm² cloth. At 1,240 nmole/cm² cloth, callicarpenal and intermedeol repelled 20 and 40% of A. americanum nymphs.     

The parasitism of immature stages of <Emphasis Type="Italic">Ixodes loricatus</Emphasis> (Acari: Ixodidae) on wild rodents in Argentina

Ixodes loricatus has long been considered with strict-total specificity to New World Marsupials. However, frequent findings of its immature stages on rodents suggest that these vertebrates play an important role in the tick’s life cycle. Aspects dealing with the ecology of Sigmodontinae rodents infestation by I. loricatus are unknown. To contribute to the knowledge of the ecology of this tick species, environmental factors, as well as host species, sex and age, were evaluated to find associations of immature I. loricatus infestation of the most abundant wild rodent species from riparian locations of Buenos Aires province (Argentina). A total of 290 hosts belonging to the subfamily Sigmodontinae were captured and examined for ticks at six locations. The data analysis showed that there was a preference of I. loricatus towards Akodon azarae, but only for those hosts captured in lands not prone to flooding. The results suggest that the host preference of I. loricatus is mainly related to factors such as habitat type and host density.     

<Emphasis Type="Italic">Manayunkia speciosa</Emphasis> Leidy (Polychaeta: Sabellidae): introduction of this nonindigenous species in the Neotropical Region (Uruguay river,South America)

We report the migration of Manayunkia speciosa from its distribution in North America into the Neotropical Region (Argentina). We collected specimens from November 2007 to March 2009 in the lower Uruguay River-at 33° 5.01′S 58° 12′W, 33° 5.9′S 58°25.2′W from sediments reaching densities of 2,890 ind. m⁻², at a mean abundance of 350 ind. m⁻². Introductions of nonindigenous species, resulting intentionally or accidentally from anthropic activities, cause significant changes in ecosystems. In aquatic environments, polychaetes are a key invasive group that increases the geographical range of several species through human activities. M. speciosa may have reached the Río de la Plata Basin through a shipping vector and thereafter the Uruguay River by self-navigation.     

Co-feeding as a route for transmission of <Emphasis Type="Italic">Rickettsia conorii israelensis</Emphasis> between <Emphasis Type="Italic">Rhipicephalus sanguineus</Emphasis> ticks

The essential oil from Amyris balsamifera (Rutaceae) and elemol, a principal constituent of the essential oil of Osage orange, Maclura pomifera (Moraceae) were evaluated in in vitro and in vivo laboratory bioassays for repellent activity against host-seeking nymphs of the blacklegged tick, Ixodes scapularis, and the lone star tick, Amblyomma americanum. Both bioassays took advantage of the tendency of these host-seeking ticks to climb slender vertical surfaces. In one bioassay, the central portion of a vertical strip of filter paper was treated with test solution and ticks placed or allowed to crawl onto the untreated lower portion. In the other bioassay, a strip of organdy cloth treated with test solution was doubly wrapped (treatment on outer layer) around the middle phalanx of a forefinger and ticks released on the fingertip. Both amyris oil and elemol were repellent to both species of ticks. Elemol did not differ significantly in effectiveness against A. americanum from the widely used repellent deet. At 2 and 4 h after application to filter paper, 827 μg amyris oil/cm² paper repelled 80 and 55%, respectively, of A. americanum nymphs. Ixodes scapularis was repelled by lower concentrations of amyris oil and elemol than A. americanum.     

Hosts,distribution and genetic divergence (16S rDNA) of <Emphasis Type="Italic">Amblyomma dubitatum</Emphasis> (Acari: Ixodidae)

We supply information about hosts and distribution of Amblyomma dubitatum. In addition, we carry out an analysis of genetic divergence among specimens of A. dubitatum from different localities and with respect to other Neotropical Amblyomma species, using sequences of 16S rDNA gene. Although specimens of A. dubitatum were collected on several mammal species as cattle horse, Tapirus terrestris, Mazama gouazoubira, Tayassu pecari, Sus scrofa, Cerdocyon thous, Myocastor coypus, Allouata caraya, Glossophaga soricina and man, most records of immature and adult stages of A. dubitatum were made on Hydrochoerus hydrochaeris, making this rodent the principal host for all parasitic stages of this ticks. Cricetidae rodents (Lundomys molitor, Scapteromys tumidus), opossums (Didelphis albiventris) and vizcacha (Lagostomus maximus) also were recorded as hosts for immature stages. All findings of A. dubitatum correspond to localities of Argentina, Brazil, Paraguay and Uruguay, and they were concentrated in the Biogeographical provinces of Pampa, Chaco, Cerrado, Brazilian Atlantic Forest, Parana Forest and Araucaria angustifolia Forest. The distribution of A. dubitatum is narrower than that of its principal host, therefore environmental variables rather than hosts determine the distributional ranges of this tick. The intraspecific genetic divergence among 16S rDNA sequences of A. dubitatum ticks collected in different localities from Argentina, Brazil and Uruguay was in all cases lower than 0.8%, whereas the differences with the remaining Amblyomma species included in the analysis were always bigger than 6.8%. Thus, the taxonomic status of A. dubitatum along its distribution appears to be certain at the specific level.     

Setal variability of <Emphasis Type="Italic">Hydrozetes lemnae</Emphasis> and <Emphasis Type="Italic">H. thienemanni</Emphasis> (Acari: Oribatida: Hydrozetidae)

Setal variability and the other morphological characters of juvenile stages and adult of Hydrozetes lemnae (Coggi, 1897) and H. thienemanni Strenzke, 1943 were investigated. In the juveniles of both species the length and shape of some setae vary, especially in the gastronotic region, more so in H. lemnae, which is parthenogenetic and reproduces by thelytoky, than in H. thienemanni, which is dioecious. The former species usually has more thick setae in the anterior and medial regions of the gastronotum, especially in the larva, compared to the latter. In the juveniles of H. lemnae the prodorsal setae le and in are longer, and in the nymphs the gastronotal setae of the l-series are usually longer than in H. thienemanni. These species differ distinctly by the number of long setae in the posterior part of the nymphs; in H. lemnae three pairs of long setae occur, while in H. thienemanni only two pairs; the respective juvenile stages of H. lemnae are also smaller than those of H. thienemanni. In the adults the number of c-series setae varies, as seta c ₁ is lost, and in some individuals also seta c ₃, and only seta c ₂ remains.     

Combined infection of<Emphasis Type="Italic">Ixodes ricinus</Emphasis> with three<Emphasis Type="Italic">Borrelia burgdorferi</Emphasis> sensu lato genotypes

Ixodes ricinus ticks were collected by random collections from western and central Slovakia during the years 1996-98. The midgut content of 240 ticks was examined by dark-field microscopy and submitted for cultivation for the presence of borrelias. Spirochetes were found in 21 unfed and host-seeking adults and nymphs (8.8%). By the analysis of restriction fragment length polymorphism (RFLP) one sample from unfed I. ricinus male from western Slovakia was identified as a triple infection of Borrelia burgdorferi sensu stricto, B. garinii and B. afzelii. The simultaneous presence of different B. burgdorferi genospecies in one midgut sample (triple infection in the tick) could be observed only after the multipart amplification of denaturated DNA and subsequent pooling of the products for further analysis.     

Associations Between Coinfection Prevalence of <Emphasis Type="Italic">Borrelia lusitaniae</Emphasis>, <Emphasis Type="Italic">Anaplasma</Emphasis> sp., and <Emphasis Type="Italic">Rickettsia</Emphasis> sp. in Hard Ticks Feeding on Reptile Hosts

An increasing number of studies reveal that ticks and their hosts are infected with multiple pathogens, suggesting that coinfection might be frequent for both vectors and wild reservoir hosts. Whereas the examination of associations between coinfecting pathogen agents in natural host–vector–pathogen systems is a prerequisite for a better understanding of disease maintenance and transmission, the associations between pathogens within vectors or hosts are seldom explicitly examined. We examined the prevalence of pathogen agents and the patterns of associations between them under natural conditions, using a previously unexamined host–vector–pathogen system—green lizards Lacerta viridis, hard ticks Ixodes ricinus, and Borrelia, Anaplasma, and Rickettsia pathogens. We found that immature ticks infesting a temperate lizard species in Central Europe were infected with multiple pathogens. Considering I. ricinus nymphs and larvae, the prevalence of Anaplasma, Borrelia, and Rickettsia was 13.1% and 8.7%, 12.8% and 1.3%, and 4.5% and 2.7%, respectively. The patterns of pathogen prevalence and observed coinfection rates suggest that the risk of tick infection with one pathogen is not independent of other pathogens. Our results indicate that Anaplasma can play a role in suppressing the transmission of Borrelia to tick vectors. Overall, however, positive effects of Borrelia on Anaplasma seem to prevail as judged by higher-than-expected Borrelia–Anaplasma coinfection rates.     

Repellency of methyl jasmonate to <Emphasis Type="Italic">Ixodes ricinus</Emphasis> nymphs (Acari: Ixodidae)

In our search for tick repellents of plant origin, to be used as alternatives to commercial arthropod repellents, we investigated the effect of the well known plant signaling compound methyl jasmonate (MJ) using nymphs of the tick Ixodes ricinus. In laboratory tests, pieces of cloth with MJ at 0.075, 0.15, 0.30 and 0.75 mg/cm² yielded increasing repellencies against the nymphs: 57%, 71%, 92% and 99%, respectively, of the nymphs did not cling to the cloth. Repellency of MJ was also investigated in a tick-infested woodland area in central Sweden. Cotton flannel cloths sprayed with 0.05, 0.1 or 0.2 mg/cm² MJ dissolved in acetone were dragged over the ground vegetation. The numbers of nymphs on the treated cloths were significantly lower than those on the untreated cloth. Thus, MJ has, at the concentrations tested, significant repellent activity against I. ricinus nymphs.     

The First Data on the <Emphasis Type="Italic">TROSPA</Emphasis> Gene Structure in <Emphasis Type="Italic">Ixodes persulcatus</Emphasis> and <Emphasis Type="Italic">Ixodes ricinus</Emphasis> Ticks from Russia

For the first time, the sequences of the TROSPA gene from taiga tick and sheep tick from Russia were obtained. Three specimens of sheep tick from Voronezh oblast and three specimens each of taiga tick from Irkutsk oblast and Perm krai were analyzed. At the end of the intron of the TROSPA gene in Ixodes persulcatus, a poly-T region was found. In addition, a fragment of the first exon containing a large number of differences in the nucleotide and amino acid composition both among species and within them was identified.     

<Emphasis Type="Italic">Asthenes dorbignyi</Emphasis> (Passeriformes: Furnariidae) host of <Emphasis Type="Italic">Argas neghmei</Emphasis> (Acari: Argasidae)

Argas neghmei Kohls & Hoogstraal is a Neotropical tick species parasite of poultry and man in the arid western of the southern cone of America in Argentina and Chile. Males, females and nymphs of an argasid were collected from 20 to 22 of June, 2007 in nests of Creamy-breasted Canastero, Asthenes dorbignyi (Reinchenbach) (Passeriformes: Furnariidae) at about 7 km S of La Poma (24°46′S, 66°12′W) and a male tick at 24°48′S, 66°10′W on Nov 27, 2008, Salta province, Argentina. Scanning electron microscope micrographies and sequences of 16S rDNA confirmed that the ticks found were A. neghmei. Development in breeding areas of A. dorbignyi is probably a source of infestation for animal premises and houses. However, the role of other unknown wild birds may also contribute to the invasion of A. neghmei in chicken houses or residences.