Morfologia e desenvolvimento do sistema reprodutor masculino de carrapatos do gênero Amblyomma (Acari, Ixodidae): uma análise comparativa

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Experimental infection of the tick Amblyomma cajennense, Cayenne tick, with Rickettsia rickettsii, the agent of Rocky Mountain spotted fever

In the laboratory, Amblyomma cajennense (Acari: Ixodidae) (Fabricius) larvae, nymphs and adults were exposed to Rickettsia rickettsii by feeding on needle-inoculated animals, and thereafter reared on uninfected guinea pigs or rabbits. Regardless of the tick stage that acquired the infection, subsequent tick stages were shown to be infected (confirming transstadial and transovarial transmissions) and were able to transmit R. rickettsii to uninfected animals, as demonstrated by serological and molecular analyses. However, the larval, nymphal and adult stages of A. cajennense were shown to be partially refractory to R. rickettsii infection, as in all cases, only part of the ticks became infected by this agent, after being exposed to rickettsemic animals. In addition, less than 50% of the infected engorged females transmitted rickettsiae transovarially, and when they did so, only part of the offspring became infected, indicating that vertical transmission alone is not enough to maintain R. rickettsii in A. cajennense for multiple generations. Finally, the R. rickettsii-infected tick groups had lower reproductive performance than the uninfected control group. Our results indicate that A. cajennense have a low efficiency to maintain R. rickettsii for successive generations, as R. rickettsii-infection rates should decline drastically throughout the successive tick generations.

Redescription of the larva of Amblyomma oblongoguttatum Koch, 1844 (Acari: Ixodidae) by light and scanning electron microscopy

The larval stage of Amblyomma oblongoguttatum Koch is redescribed using optical and scanning electron microscopy. Unfed larvae were obtained from a colony of A. oblongoguttatum originated from engorged females collected on domestic pigs from Monte Negro municipally (10 degrees 29'S, 63 degrees 32'W), State of Rondonia, Western Amazon, Brazil. Several characters are presented including the chaetotaxy of the idiosoma, palpi and Haller's organ, as well as morphological features of the idiosoma, gnathosoma and legs. In addition, the porotaxy (topographical and numerical patterns of integumentary structures) were presented by using a new nomenclature recently proposed. The chaetotaxy of the larvae of A. oblongoguttatum, in general, is similar to other Neotropical Amblyomma species. Three types of integumentary structures were observed on the idiosoma: lyrifissures, small glands, and large wax glands. Topographic and numerical patterns of the integumentary structures consisted of 5 pairs of large wax glands (1 dorsal/4 ventral), 24 pairs of lyrifissures (11 dorsal/13 ventral), and 49 pairs of small glands (28 dorsal/21 ventral). These topographic and numerical patterns found for A. oblongoguttatum show only minor differences when compared with patterns of other Amblyomma larvae, however, a few key features can be used for identification of these species.

Coxiella symbiont in the tick Ornithodoros rostratus (Acari: Argasidae)

In the present study, the presence of tick-associated bacteria and protozoa in Ornithodoros rostratus ticks (adults, nymphs, and eggs) from the Pantanal region of Brazil were determined by molecular detection. In these ticks, DNA from protozoa in the genera Babesia and Hepatozoon, and bacteria from the genera Rickettsia, Borrelia, Anaplasma, and Ehrlichia were not detected. Conversely, all tested ticks (100%) yielded PCR products for 3 Coxiella genes (16S rRNA, pyrG, cap). PCR and phylogenetic analysis of 3 amplified genes (16S rRNA, pyrG, cap) demonstrated that the agent infecting O. rostratus ticks was a member of the genus Coxiella. This organism grouped with Coxiella symbionts of other soft tick species (Argasidae), having different isolates of C. burnetii as a sister group, and these 2 groups formed a clade that grouped with another clade containing Coxiella symbionts of hard tick species (Ixodidae). Analysis of tick mitochondrial 16S rRNA gene database composed mostly of tick species previously shown to harbor Coxiella symbionts suggests a phylogenetic congruence of ticks and their Coxiella symbionts. Furthermore, these results suggest a very long period of coevolution between ticks and Coxiella symbionts and indicates that the original infection may have occurred in an ancestor common to the 2 main tick families, Argasidae (soft ticks) and Ixodidae (hard ticks). However, this evolutionary relationship must be confirmed by more extensive testing of additional tick species and expanded populations. (c) 2012 Elsevier GmbH. All rights reserved.

A surrogate life cycle of Amblyomma ovale Koch, 1844

As ticks spend most of their time off-host, the environment is a major selective force of these parasites. In fact, human impact on landscapes has favored a minority of tick species which became well-known pests. However, this is an ongoing process and novel pests may arise. We herein report a surrogate life cycle of a neotropical tick species. Amblyomma ovate, and which may be related to an increased risk of human rickettsiosis. Under natural conditions, adults of this tick species feed on carnivores and exhibit non-nidicolous ambush behavior, whereas larvae and nymphs feed on small rodents and birds. In an anthropized spot within an Atlantic rainforest reserve of Brazil, an A. ovate population exhibited a nidicolous behavior with all 3 tick stages feeding on the dog. This dog's infestation was outstandingly high, and it displayed the highest anti-Rickettsia titers and harbored Rickettsia-infected ticks. (c) 2012 Elsevier GmbH. All rights reserved.

Brazilian spotted fever: Real-time PCR for diagnosis of fatal cases

Suspicion of Brazilian spotted fever (BSF) should occur in endemic regions upon surveillance of the acute febrile icteric hemorrhagic syndrome (AFIHS). However, limitations associated with currently available laboratory tests pose a challenge to early diagnosis, especially in fatal cases. Two real-time PCR (qPCR) protocols were evaluated to diagnose BSF in 110 fatal AFIHS cases, collected in BSF-endemic regions in 2009-2010. Of these, 24 were positive and 86 negative by indirect immunofluorescence (IFA) assay (cutoff IgG and/or IgM >= 128). DNA from these samples was used in the qPCR protocols: one to detect Rickettsia spp. (Citrate synthase gene) and another to determine spotted fever group (SFG) Rickettsia species (OmpA gene). Of the 24 IFA-positive samples, 5 (21%) were positive for OmpA and 9 (38%) for citrate synthase. In the IFA-negative group (n = 86), OmpA and citrate synthase were positive in 23 (27%) and 27 (31%), respectively. These results showed that the 2 qPCR protocols were about twice as sensitive as the IFA test alone (93% concordance). In conclusion, qPCR is a sensitive method for the diagnosis of fatal BSF cases and should be considered for routine surveillance of AFIHS in places like Brazil, where spotted fever-related lethality is high and other endemic diseases like dengue and leptospirosis can mislead diagnosis. (C) 2012 Elsevier GmbH. All rights reserved.

Morphological records of oocyte maturation in the parthenogenetic tick Amblyomma rotundatum Koch, 1844 (Acari: Ixodidae)

Oocyte maturation in the thelytokous parthenogenetic tick Amblyomma rotundatum was examined for the first time using light and scanning electron microscopy. The panoistic ovary lacks nurse and follicular cells and is a single continuous tubular structure forming a lumen delimited by the ovarian wall. Oocytes of tick species are usually classified according to cytoplasm appearance, the presence of germinal vesicle, the presence of yolk granules, and the chorion. However, for this species, we also use oocyte size as an auxiliary tool since most oocytes were in stages I-Ill and were histologically very similar. Oocytes were classified into five development stages, and specific characteristics were observed: mature oocytes with thin chorion, pedicel cells arranged forming an epithelium with two Or more oocytes attached by the same structure, and a large number of oocytes in the process of reabsorption. (C) 2011 Elsevier GmbH. All rights reserved.

A Third Amblyomma Species and the First Tick-Borne Rickettsia in Chile

During November 2010, three ticks were collected from three dogs living in the rural area of Arica, northern Chile. Morphological analyses of the ticks in the laboratory revealed that they were most similar to Amblyomma maculatum Koch and Amblyomma triste Koch. However, because of unique metatarsal spurs, neither of the Chilean specimens could be assigned with certainty to A. maculatum or A. triste, based on external morphology. The mitochondrial 16S rRNA gene partial sequences obtained from two Chilean specimens were 99.5% identical to A. triste from Uruguay, and 99.0% identical to A. maculatum from the United States. Through phylogenetic analysis inferred from partial 16S rRNA sequences, the Chilean specimens were classified as A. triste. Molecular analyses also showed that one of the three Chilean ticks was infected by Candidatus 'Rickettsia andeanae'. These findings extend the geographical distribution of A. triste to Chile, where no tick-associated rickettsia had been reported previously.

CLEANING ASSOCIATIONS BETWEEN BIRDS AND HERBIVOROUS MAMMALS IN BRAZIL: STRUCTURE AND COMPLEXITY

Birds that remove ectoparasites and other food material from their hosts are iconic illustrations of mutualistic-commensalistic cleaning associations. To assess the complex pattern of food resource use embedded in cleaning interactions of an assemblage of birds and their herbivorous mammal hosts in open habitats in Brazil, we used a network approach that characterized their patterns of association. Cleaning interactions showed a distinctly nested pattern, related to the number of interactions of cleaners and hosts and to the range of food types that each host species provided. Hosts that provided a wide range of food types (flies, ticks, tissue and blood, and organic debris) were attended by more species of cleaners and formed the core of the web. On the other hand, core cleaner species did not exploit the full range of available food resources, but used a variety of host species to exploit these resources instead. The structure that we found indicates that cleaners rely on cleaning interactions to obtain food types that would not be available otherwise (e.g., blood-engorged ticks or horseflies, wounded tissue). Additionally, a nested organization for the cleaner bird mammalian herbivore association means that both generalist and selective species take part in the interactions and that partners of selective species form an ordered subset of the partners of generalist species. The availability of predictable protein-rich food sources for birds provided by cleaning interactions may lead to an evolutionary pathway favoring their increased use by birds that forage opportunistically. Received 30 June 2011, accepted 10 November 2011.

Epidemiology of Brazilian spotted fever in the Atlantic Forest, state of Sao Paulo, Brazil

The tick-borne bacterium Rickettsia rickettsii is the aetiological agent of Brazilian spotted fever (BSF). The present study evaluated tick infestations on wild and domestic animals, and the rickettsial infection in these animals and their ticks in 7 forest areas adjacent to human communities in the Sao Paulo Metropolitan Area (SPMA). The results were compared to ecological traits of each sampled area. Two main tick species, Amblyomma aureolatum and Rhipicephalus sanguineus, were collected from dogs. The major ticks found on small mammals and birds were Ixodes loricatus and Amblyomma longirostre, respectively. Both anti-R. rickettsii antibodies and R. rickettsii-infected ticks were detected on dogs from only 2 areas in the southern part of the SPMA, which were considered to be endemic for BSF; the remaining 5 areas were considered to be non-endemic. Ecologically, the BSF-endemic areas clearly differed from the non-endemic areas by the presence of significantly more degraded forest patches in the former. The present results corroborate historical observations that have indicated that all human cases of BSF in the SPMA were contracted in the southern part of this metropolitan area. However, not all forest patches in the southern part of the SPMA were shown to be associated with BSF endemism.

Maternal Care in the Soft Tick Antricola marginatus

Among spiders, scorpions, and whip spiders, a common type of maternal care consists of females carrying newly hatched offspring on their body for a few days until they are able to live independently. While this maternal care has been suggested to occur in different argasid tick species, it has been recorded only once, for Antricola marginatus in Cuba; however, this earlier record only superficially mentioned the occurrence of this behavior, with no further details. Here we report the occurrence of maternal care in the argasid tick A. marginatus under natural conditions in a cave at Yucatan. Mexico, where 8 A. marginatus females, while walking on bat guano, had their body entirely covered by a mean number of 305 +/- 112 conspecific unfed larvae (range: 105-466). Larvae covered the entire idiosoma of the female tick, where they were motionless or displayed just slight movement. This result substantially expands the number of unique characters that have been found only in Antricola spp, ticks, when compared to the other tick genera. Our findings also indicate that maternal care evolved independently in different taxa of Arachnida, since it has been reported for species of Araneae, Scorpiones, and Amblypygi, and here for an Acari species.

Ecological implications on the aggregation of Amblyomma fuscum (Acari: Ixodidae) on Thrichomys laurentius (Rodentia: Echimyidae), in northeastern Brazil

We investigated the Amblyomma fuscum load on a pullulating wild rodent population and the environmental and biological factors influencing the tick load on the hosts. One hundred and three individuals of Thrichomys laurentius were caught in an Atlantic forest fragment in northeastern Brazil, as part of a longitudinal survey on ticks infesting non-volant small mammals. Ticks (n = 342) were found on 45 individuals and the overall mean intensity of infestation was 7.6 ticks per infested rodent. Ticks were highly aggregated in the host population and the negative binomial distribution model provides a statistically satisfactory fit. The aggregated distribution was influenced by sex and age of the host. The microhabitat preference by T. laurentius probably increases contact opportunities between hosts and aggregated infesting stages of the ticks and represents important clues about the habitat suitability for A. fuscum.

Life-cycle and host preference of Amblyomma ovale (Acari: Ixodidae) under laboratory conditions

This study evaluated for the first time the life cycle of Amblyomma ovale in the laboratory. For this purpose, larvae and nymphs were exposed to Gallus gallus (chickens), Cavia porcellus (guinea pigs), Rattus norvegicus (wistar rats), Oryctolagus cuniculus (domestic rabbits), Calomys callosus (vesper mouse), and Didelphis albiventris (white-eared opossum). Nymphs were also exposed to Nectomys squamipes (South American water rat). Adult ticks were fed on dogs. The life-cycle of A. ovale in laboratory could be completed in an average period of ca. 190 days, considering prefeeding periods of 30 days for each of the parasitic stages. Vesper mice were the most suitable host for A. ovale larvae, whereas water rats were the most suitable host for A. ovale nymphs. Our results, coupled with literature data, strongly indicate that small rodents have an important role in the life history of A. ovale. Chickens (the only avian host used in the present study) showed to be moderately suitable hosts for subadult A. ovale ticks, indicating that wild birds might have a secondary role in the life history of A. ovale. Domestic dogs showed to be highly suitable for the adult stage of A. ovale, in agreement with literature data that indicate that the domestic dog is currently one of the most important hosts of A. ovale adult ticks in Latin America.

Distribution of Rickettsia rickettsii in ovary cells of Rhipicephalus sanguineus (Latreille1806) (Acari: Ixodidae)

Abstract Background Considering the fact that the dog tick, Rhipicephalus sanguineus, has a great potential to become the vector of Brazilian Spotted Fever (BSF) for humans, the present study aimed to describe the distribution of the bacterium Rickettsia rickettsii, the etiological agent of BSF, in different regions of the ovaries of R. sanguineus using histological techniques. The ovaries were obtained from positive females confirmed by the hemolymph test and fed in the nymph stage on guinea pigs inoculated with R. rickettsii. Results The results showed a general distribution of R. rickettsii in the ovary cells, being found in oocytes in all stages of development (I, II, III, IV and V) most commonly in the periphery of the oocyte and also in the cytoplasm of pedicel cells. Conclusions The histological analysis of the ovaries of R. sanguineus infected females confirmed the presence of the bacterium, indicating that the infection can interfere negatively in the process of reproduction of the ticks, once alterations were detected both in the shape and cell structure of the oocytes which contained bacteria.

Ornithodoros peropteryx (Acari: Argasidae) in Bolivia: an argasid tick with a single nymphal stage

By the end of the 1960s, the argasid tick Ornithodoros peropteryx was described from larval specimens collected from the bat Peropteryx macrotis in Colombia. Since its original description, no additional record of O. peropteryx has been reported, and its post-larval stages have remained unknown. During July 2010, 18 larvae were collected from 9 bats (Centronycteris maximiliani), resulting in a mean infestation of 2.0 ± 2.2 ticks per bat (range 1–8). These bats were captured in a farm in northeastern Bolivia close to Guaporé River in the border with Brazil. Morphological examinations of the larvae revealed them to represent the species O. peropteryx. One engorged larva that was kept alive in the laboratory moulted to a nymph after 9 days. Fourteen days after the larval moulting, the nymph moulted to an adult female without taking any blood meal during the nymphal period. This adult female was used for a morphological description of the female stage of O. peropteryx. In addition, the larvae were used for a morphological redescription of this stage. One larva and two legs extirpated from the adult female were submitted to DNA extraction and PCR targeting a fragment of the mitochondrial 16S rDNA gene, which yielded DNA sequences at least 11 % divergent from any available argasid sequence in Genbank. We show that O. peropteryx ontogeny is characterized by a single, non-feeding, nymphal stage. This condition has never been reported for ticks.