Rickettsia parkeri strain Atlantic rainforest infecting Amblyomma ovale (Acari: Ixodidae) in the Amazon Biome (Acre state,Brazil)

There is a lack of studies regarding tick-associated Rickettsia in the Amazon biome. Aiming to contribute to this knowledge, our research group collected ticks in the Western Amazon to better understand the tick fauna and their associated Rickettsia. In this study, we detected Rickettsia parkeri strain Atlantic rainforest DNA in the tick Amblyomma ovale Koch, 1844 in Rio Branco municipality, Acre state, northern Brazil. This is the first time that the R. parkeri strain Atlantic rainforest has been reported in the Amazon biome and is the first evidence of the circulation of a pathogenic spotted fever group (SFG) Rickettsia in this biome. This finding provides substantial information to help public health authorities understand which species of Rickettsia may be related to Amazon spotted fever cases.     

Tick fauna in non-anthropogenic areas in Mato Grosso do Sul,Brazil, with the presence of the Rickettsia parkeri strain Atlantic rainforest in Amblyomma ovale

The aim of this study was to evaluate tick fauna and identify the possible presence of Rickettsia spp. in ticks of the genus Amblyomma from two environmental preservation areas in different regions of Mato Grosso do Sul state, Brazil. CO₂ traps, visual observation and cloth dragging were used to capture ticks. Three hundred ticks were submitted to the hemolymph test, and samples that showed organisms morphologically compatible with Rickettsia were used for rickettsial DNA detection by PCR. DNA was extracted using guanidine-phenol isothiocyanate, and the primers CS78 and CS323 were used for PCR, which amplified a 401-base pair fragment of the citrate synthase (gltA) gene. If positive, the DNA sample was tested by primers Rr190.70p and Rr190.602n that produce a 530 bp amplicon of the ompA gene that is present only in rickettsiae of the spotted fever group. A total of 1,745 adult ticks were collected, including 1,673 specimens of Amblyomma sculptum, 63 of Amblyomma coelebs, five of Amblyomma naponense and four of Amblyomma ovale. Thirteen ticks of the species A. ovale, A. coelebs and A. sculptum showed structures compatible with Rickettsia inside the hemocytes; after DNA extraction, the presence of Rickettsia spp. in a sample of A. ovale was confirmed by PCR in both analyzed fragments. In the sequencing analysis, 100% identity for the Rickettsia parkeri strain Atlantic rainforest was obtained according to GenBank. The two environmental preservation areas showed A. sculptum as the predominant species, as well as the presence of marked seasonality for this species. This paper is the first report of the R. parkeri strain Atlantic rainforest in A. ovale ticks in the state of Mato Grosso do Sul, Brazil.     

Epidemiological aspects of Rickettsia parkeri in the Atlantic forest biome of Espírito Santo state,Brazil

Since 2010, a new rickettsiosis caused by Rickettsia parkeri, a species included in the spotted fever group (SFG) and transmitted by the tick Amblyomma ovale, has been described in Brazil. Considering that A. ovale does occur in the Espírito Santo state (ES), the present study aimed at preliminarily describing the epidemiology of R. parkeri in the Atlantic forest of this state. Between June 2016 and September 2018, 33 villages from nine municipalities of the ES were included in a study for the molecular detection and isolation of SFG rickettsiae from adult Amblyomma ticks collected on dogs with free access to the forest. Serologic screenings against SFG rickettsiae in these animals and their owners (humans) were performed through immunofluorescence assay (IFA) using antigens of Rickettsia rickettsii and R. parkeri. Additionally, local health secretariats were informed on clinical manifestations of R. parkeri infection and told to communicate any suspected case. A total of 280 adult ticks were collected and taxonomically classified as A. ovale (n = 152), Amblyomma aureolatum (n = 127) and Amblyomma sculptum (n = 1). Overall, Rickettsia DNA was detected in 12/266 ticks. The sequencing of PCR products revealed that 0.7% (1/144) and 0.8% (1/121) of the analyzed A. ovale and A. aureolatum ticks were infected by R. parkeri strain Atlantic rainforest, respectively, and 8.3% (10/121) of the A. aureolatum ticks infected by Rickettsia bellii. Among the above PCR-positive ticks, only one isolate from one A. aureolatum tick was successfully established in the laboratory. DNA extracted from the third passage of this isolate was designated as strain M9A and molecularly characterized using primers targeting the Rickettsia gltA gene, whose sequence matched 100% the corresponding sequences of R. bellii. Seroprevalence against SFG rickettsiae in sampled dogs (n = 83) was 41% or 57%, depending on the rickettsial antigen (R. rickettsii strain Taiaçu or R. parkeri strain Atlantic rainforest, respectively). A total of 37 (45%) canine sera showed titers to R. parkeri at least fourfold higher than to R. rickettsii antigen. Among humans, 10% (4/41) of the samples reacted to at least one rickettsial antigen, with the highest endpoint titer varying from 64 to 128 for R. rickettsii and R. parkeri; no human serum showed ≥4-fold difference between the highest endpoint titers. Finally, during the study period, suspicions on cases of R. parkeri-rickettsiosis were not informed by the health secretariats. Our results confirm the presence and exposure to R. parkeri strain Atlantic rainforest, associated with two anthropophilic tick species (A. ovale and A. aureolatum) parasitizing domestic dogs with unrestrained access to forest areas. Consequently, the occurrence of R. parkeri infection in humans inhabiting the Atlantic forests of ES should not be discarded.     

Records of ticks (Acari: Ixodidae) on humans and distribution of spotted-fever cases and its tick vectors in Paraná State,southern Brazil

Brazilian spotted fever (BSF) is the deadliest rickettsiosis in the world. Although the epidemiology of the disease has been established in Brazil, there are still limited data available on distribution of tick vectors and tick species parasitizing humans in the country, particularly in Paraná State. The State of Paraná is located in the southern region of the country and is covered by two biomes: Atlantic rainforest and Cerrado. Thus, the aims of this study were i) to map the distribution of SF tick vectors, ii) to describe and map the distribution of human parasitism by ticks, and iii) to map the distribution of fatal and non-fatal spotted fever (SF) cases in Paraná State, southern Brazil. Data were reviewed and compiled from previous published reports, and also from two scientific collections of Paraná State. SF cases were retrieved from the Brazilian Notifiable Diseases Information System. A total of 50 cases of human parasitism by ticks were recorded, with a total of 64 (22 males, 12 females, 30 nymphs) ticks collected. The following 12 tick species were identified: Amblyomma aureolatum, Amblyomma brasiliense, Amblyomma calcaratum, Amblyomma dubitatum, Amblyomma incisum, Amblyomma longirostre, Amblyomma ovale, Amblyomma parkeri, Amblyomma scalpturatum, Amblyomma sculptum, Haemaphysalis juxtakochi and Rhipicephalus sanguineus sensu lato (s.l.). The most prevalent tick species associated to cases of human parasitism were A. sculptum (13/50; 26 %), A. aureolatum (10/50; 20 %), A. brasiliense (5/50; 10 %), A. ovale (5/50; 10 %) and A. parkeri (4/50; 8%). A total of 51 non-fatal and five fatal SF cases were recorded. Data from this study highlights the need for monitoring ticks parasitizing humans aiming early detection of tick-borne diseases cases, particularly BSF in Paraná State, southern Brazil.     

The inoculation eschar of Rickettsia parkeri rickettsiosis in Brazil: Importance and cautions

Two well characterized tick-borne rickettsioses occur in Brazil. Rickettsia rickettsii caused spotted-fever, transmitted by Amblyomma sculptum and Amblyomma aureolatum, is a severe disease with a high case-fatality rate in the southeastern region of the country. Rickettsia parkeri strain Atlantic rainforest infections transmitted by adult Amblyomma ovale ticks cause a milder non-lethal febrile disease with an eschar (necrosis) at the tick bite site. Clinical diagnosis of rickettsiosis is challenging, particularly during the early stages of the illness when signs and symptoms are non-specific. Since eschar at the tick bite site has emerged as the main clinical feature of mild R. parkeri infections and used to differentiate it from severe R. rickettsii infection, its proper recognition, distinction from other tick bite lesions, and boundaries as a clinical tool must be highlighted. Of importance, eschars induced by Rickettsia must be differentiated from dermatoses caused by other tick-borne skin infections as well from lesions caused by the tick bite itself. We herein highlight information on eschar in rickettsial diseases in Brazil and discuss the need for further research on its clinical relevance and application in the diagnosis of spotted fever caused by R. parkeri strain Atlantic rainforest. In particular, we draw attention to diagnosis of other febrile diseases in the presence of concomitant tick bites.     

Detection of Rickettsia tamurae-like and other spotted fever group rickettsiae in ticks (Acari: Ixodidae) associated with wild birds in the Western Amazon,Brazil

Ticks are vectors for several pathogens, including bacteria belonging to the Rickettsia genus, such as Rickettsia rickettsii and Rickettsia parkeri, the causative agents of spotted fever. The aim of the present study was to investigate the tick species richness and rickettsial agents associated with wild birds captured in the Humaita Forest Reserve, Acre, in the Western Amazon region. Wild birds were captured with ornithological nets for visual inspection with the purpose of collecting ticks, which were identified through morphological analyses and molecular tests for several genes (12S rDNA, 16S rDNA, gltA, ompA, and sca4). A total of 607 wild birds were captured, 12% of which were parasitized by 268 ticks of the Amblyomma genus, with new host-parasite associations reported for Amblyomma calcaratum, Amblyomma geayi, Amblyomma longirostre, Amblyomma naponense, Amblyomma nodosum, and Amblyomma varium. Of the total ticks collected, 113 were tested for the presence of rickettsial DNA fragments, with 19 testing positive for R. parkeri in A. geayi, Rickettsia tamurae-like in Amblyomma sp., and Rickettsia amblyommatis in A. geayi, A. longirostre, and Amblyomma sp. We detected R. tamurae-like in Amblyomma larvae for the first time in the Western Brazilian Amazon biome, and registered spotted fever group rickettsiae, although the relevance of the detected species in a public health context should be further explored in South America, as well as new host-parasite interactions in this underexplored region.     

Rickettsia spp. in ticks from a tropical dry forest reserve on Mexico's Pacific Coast

The present study analyzes the diversity of bacteria of the genus Rickettsia in ticks collected from wild mammals and vegetation at the Chamela Biological Station (EBCh), Jalisco, Mexico, a core area in the Chamela-Cuixmala Biosphere Reserve, which is characterized by the presence of tropical dry forest, one of the most threatened ecosystems in the world. Ticks were collected during April and November 2018 and 2019, determined to species, and subsequently processed for rickettsiae. Genomic DNA was extracted from each of 349 ticks (198 from mammals, 151 from vegetation) and screened for Rickettsia by targeting the gltA and ompA genes. Overall, 75 ticks (21.5%) were positive for rickettsiae (46 from mammals, 29 from vegetation). Through BLASTn and maximum likelihood analysis, four Rickettsia taxa were identified. Only Rickettsia amblyommatis had been previously recorded in the vicinity of the EBCh, and we confirm its presence in this reserve, associated with Amblyomma mixtum, Amblyomma cf. oblongoguttatum and Amblyomma parvum. Additionally, we record for the first time at EBCh the taxa Rickettsia parkeri strain Atlantic rainforest, which is known to cause spotted fever illness in humans, detected in Amblyomma ovale; Rickettsia bellii in A. parvum and Ornithodoros talaje; and an undescribed Rickettsia sp. in Ixodes sinaloa.     

Molecular detection of Rickettsia amblyommatis and Rickettsia parkeri in ticks collected from wild pigs in Campeche,Mexico

Tick-borne rickettsioses are caused at least by 15 species of Rickettsia of the Spotted fever group, which represent a major emerging and re-emerging public health problem worldwide. Some of these microorganisms have complex cycles involving the interaction of multiple species of ticks and wild and domestic mammals. Rickettsia infection was investigated in ticks collected from wild pigs at six localities in southeastern Mexico. We collected and tested 196 ticks belonging to four species, including Amblyomma maculatum, Amblyomma mixtum, Amblyomma ovale and Riphicephalus microplus, from 13 of 20 (65%) wild pigs sampled. Overall, Rickettsia DNA was detected in 13.8% of ticks tested (10 ♂ and 17 ♀). Of the 27 Rickettsia-positive ticks, six were A. maculatum, and 21 A. mixtum. Sequencing and phylogenetic analysis of the gltA and ompB genes revealed the presence of Rickettsia parkeri sensu stricto in one female A. maculatum and Rickettsia amblyommatis in five A. maculatum (2 ♂, 3 ♀) and 21 A. mixtum ticks (8 ♂, 13 ♀). The finding of two rickettsial agents in ticks collected from a wild pig population that is regularly captured and kept in captivity or hunted as a source of food raises concern about potential disease transmission to humans and domestic animals. However, more investigations are needed to further understand the ecology of Rickettsia species in free-ranging animals and their implications for human health.     

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 ovale, 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. ovale 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.     

Ticks on humans in an Atlantic rainforest preserved ecosystem in Brazil: Species,life stages,attachment sites,and temporal pattern of infestation

We herein describe exposure and tick attachment to researchers and field staff within the greatest preserved inland Atlantic rainforest, located in Brazil, over a non-consecutive period of five years. Overall, 433 ticks from five species were collected, Amblyomma coelebs (n = 281), Amblyomma brasiliense (n = 79), Amblyomma incisum (n = 43), Amblyomma ovale (n = 4), Haemaphysalis juxtakochi (n = 4). Clusters of larvae belonged to the genus Amblyomma (n = 21) and Haemaphysalis (n = 1). Both Haemaphysalis and Amblyomma species were observed crawling on humans but only ticks from the latter genus were found attached. Most recovered ticks (59%) were found attached and, whereas A. brasiliense was the main host-seeking tick species in the forest, A. coelebs nymphs were the ones that overwhelmingly attached to humans (70% of all attachments). At the same time only 0.4% of human bites were of A. ovale, the Rickettsia parkeri strain Atlantic rainforest vector tick in Brazil. Tick attachment sites were recorded for 132 ticks and lower/upper limbs and the trunk were more parasitized than the head/neck. Tick bites were recorded throughout the year; while larvae did not show an evident seasonal pattern, nymphs attached to humans mainly in autumn and winter and the few adult bites were recorded in spring. Our results highlighted A. coelebs nymphs as major human-biters and evidenced the necessity of studies that discern human tick bites that occur after exposure to host-seeking ticks from those that occur after tick displacement from hosts that carry ticks to human dwellings.     

Free-living ticks (Acari: Ixodidae) in the Iguaçu National Park,Brazil: Temporal dynamics and questing behavior on vegetation

Basic knowledge of species, ecological preferences and behavior of ticks in preserved rainforests are scanty. We herein describe species richness, seasonal activity, altitude influence and questing height of ticks along 3375 m of animal trails within the largest remain of inland Atlantic rainforest in Brazil, the Iguaçu National Park (INP). Altogether, 2954 ticks from nine tick species were collected; Amblyomma brasiliense Aragão (49.09% of the specimens), Amblyomma coelebs Neumann (29.08%), Amblyomma incisum Neumann (12.49%), Haemaphysalis juxtakochi Cooley (5.21%), Amblyomma ovale Koch (2.64%), Amblyomma longirostre Koch (0.20%), Ixodes fuscipes Fonseca (0.17%), Ixodes loricatus Neumann (0.07%) and Amblyomma dubitatum Neumann (0.03%) (Ixodida: Ixodidae). There was an evident seasonal influence on tick host-questing activity, with each tick stage prevailing in a season before the following one, and a general trend of one tick generation per year. Adult tick questing height mode on vegetation showed a correspondence between tick species and preferred host size and height. Adult ticks quested higher than immatures, but several larval clusters were found high enough to seek for large-sized hosts as well. Nymphal and adult tick questing height increased significantly at higher altitudes within the park. Amblyomma incisum and A. coelebs were more abundant at higher and lower altitudes, respectively. The knowledge on tick species richness, ecological preferences and behavior herein obtained provides baseline information to understand tick-borne disease epidemiology occurring at landscapes under a strong anthropogenic impact.     

Survey of bacterial and protozoan agents in ticks and fleas found on wild animals in the state of Rio de Janeiro,Brazil

This study evaluates the presence of bacterial and protozoan agents in ticks and fleas found on wild animals in the state of Rio de Janeiro, Brazil. These ectoparasites were collected on mammal species Hydrochoerus hydrochaeris, Tapirus terrestris, Dicotyles tajacu, Didelphis aurita, Cuniculus paca, Cerdocyon thous, and Coendou prehensilis, and on the terrestrial bird Dromaius novaehollandiae. Ticks and fleas were identified morphologically using specific taxonomic keys. A total of 396 ticks and 54 fleas were tested via polymerase chain reaction (PCR) for the presence of Rickettsia spp., Borrelia spp., microorganisms of the order Piroplasmida and Anaplasmataceae family. This total is distributed among nine tick species of the genus Amblyomma and one flea species. Rickettsia bellii was detected in Amblyomma dubitatum and Amblyomma pacae; Rickettsia sp. strain AL was found in Amblyomma longirostre; Rickettsia parkeri strain Atlantic rainforest was found in Amblyomma ovale; and “Candidatus Rickettsia senegalensis” and Rickettsia felis were detected in Ctenocephalides felis felis. Wolbachia sp. was detected in C. f. felis, and Borrelia sp. was detected in Amblyomma calcaratum (here named Borrelia sp. strain Acalc110). All tested samples were negative for Ehrlichia spp. and microorganisms of the Piroplasmida order. This study detected a new bacterial strain, Borrelia sp. strain Acalc 110 (which is genetically close to B. miyamotoi and B. venezuelensis) and the Rickettsia sp. strain 19P, which is 100% similar to “Ca. R. senegalensis”, a bacterium recently discovered and now being reported for the first time in Brazil.     

Detection of Rickettsia spp. in ticks of wildlife fauna from Costa Rica: First report of Rickettsia rhipicephali in Central America

In the past two decades, new species of Rickettsia have been detected and described worldwide, some of them considered pathogenic for humans. Although Costa Rica is considered a biodiversity hotspot, the knowledge about rickettsiae in sylvatic ecosystems and wild animals is scarce. The aim of this preliminary study was to detect and identify species of Rickettsia in ticks collected from wild animals in Costa Rica. A total 119 ticks were collected from 16 animal host species belonging to diverse vertebrate families (Didelphidae, Procyonidae, Felidae, Choloepodidae, Bradypodidae, Myrmecophagidae, Tayassuidae, Tapiridae, Phyllostomidae, Bufonidae, Geoemydidae, Boidae, Colubridae), and they were grouped into 43 pools to detect the presence of Rickettsia spp. DNA by PCR targeting the gltA gene. In positive pools, amplicons of the ompA, sca5 (ompB), and/or htrA genes were also amplified to identify the species present. The identification of some ticks was also confirmed by molecular methods. Four species of Rickettsia were detected in eight (19%) tick pools: Rickettsia amblyommatis in four pools of Amblyomma geayi (host: Caluromys derbianus) and one pool of Amblyomma cf. parvum (host: Nasua narica), Rickettsia rhipicephali in one pool of Dermacentor latus (host: Tayassu pecari), ‘Candidatus Rickettsia colombianensi’ in one pool of Amblyomma sp. nymphs (host: Boa constrictor), and Rickettsia sp. genotype IbR/CRC in one pool of Ixodes cf. boliviensis (host: Puma concolor). This is the first molecular detection of R. rhipicephali in Central America, and of ‘Candidatus R. colombianensi’ in Costa Rica. Results show that diverse wild animals and their ticks are associated with several species of rickettsiae in Costa Rica, which may come in contact with humans and other domestic animals in sylvatic environments.     

Immature hard ticks infected with Rickettsia amblyommatis on breeding birds from Pantanal

Immature hard ticks from the genus Amblyomma feed on blood from a wide range of Neotropical avian hosts. They serve as vectors for pathogens of medical and veterinary importance, such as Rickettsia agents of the spotted fever group (SFG). Hence, determining ecological factors that increase encounter rates between immature ticks and their avian hosts may contribute to the understanding of tick-borne diseases transmission. Here, we used 720 individual birds from 96 species surveyed in the Brazilian Pantanal to test whether host breeding season influenced tick infestation probabilities. Additionally, collected ticks were screened for Rickettsia agents to describe new avian-tick-bacteria associations. Our models revealed that the probability of an individual bird being infested with immature ticks was similar during the breeding and pre-breeding season, but higher loads of immature tick stages were found during the breeding season. Host sex did not predict infestation probability, but Rickettsia agents recovered from ticks were more prevalent during the pre-breeding season. The new records of host usage by larvae and nymphs of Amblyomma in Pantanal and the growing body of tick surveys in Neotropical avian communities, suggest that immature ticks may benefit from avian blood sources during their annual cycle. The low number of infected ticks with Rickettsia agents on Pantanal birds suggest that this vertebrate group are likely not acting as reservoirs for these microorganisms. However, long-term surveys at the same site are imperative to determine which tick species are acting as reservoirs for Rickettsia agents in Pantanal and determine whether birds are playing a role in dispersing ticks and tick-borne pathogens.     

Detection of Rickettsia spp. in ring-tailed coatis (Nasua nasua) and ticks of the Iguaçu National Park,Brazilian Atlantic Rainforest

Wild animals are of considerable importance in the ecology of infectious agents, as they can function as hosts and even as possible vectors. In this study, DNA from Rickettsia spp. was detected on ticks and fragments of skin collected from wild coatis with synanthropic habits in the Iguaçu National Park (INP) in the state of Paraná in southern Brazil. Testing was carried out on a total of 566 ticks, comprising Amblyomma spp. larvae, nymphs of Haemaphysalis juxtakochi, Amblyomma brasiliense, Amblyomma coelebs, and adults of Amblyomma ovale. The samples were tested by polymerase chain reaction (PCR) by amplifying htrA, gltA, ompA, and ompB gene fragments to detect Rickettsia spp. A fragment of each positive sample was sequenced in both directions, submitted to Genbank for a homology search, and also used for phylogenetic analyses. Samples of A. coelebs (1.90%, 8/420), A. ovale (13%, 6/45), and ring-tailed coati skin (1%, 1/75) amplified Rickettsia spp. DNA. Through sequencing, Rickettsia bellii was observed in A. ovale, Rickettsia amblyommatis in A. coelebs, while Rickettsia rhipicephali was detected in the skin samples. Wild ring-tailed coatis with synanthropic habits in the INP and their ticks are infected by Rickettsia spp., and associations with new hosts have been described.     

Ticks (Acari: Ixodidae,Argasidae) associated with wild birds in Argentina

The aim of this study was to report tick infestations on wild birds from four Phytogeographic Provinces of Argentina. A total of 1085 birds was captured (124 species, 97 genera, 29 families and 13 orders), and ticks were collected from 265 birds (48 species, 40 genera and five orders). A total of 1469 ticks (1102 larvae, 363 nymphs and 4 females) belonging to 15 tick species (Amblyomma calcaratum, Amblyomma dubitatum, Amblyomma nodosum, Amblyomma ovale, Amblyomma parvum, Amblyomma sculptum, Amblyomma tigrinum, Amblyomma triste, Haemaphysalis juxtakochi, Haemaphysalis leporispalustris, Ixodes auritulus sensu lato, Ixodes pararicinus, Ixodes silvanus, Ixodes sp. cf. I. affinis and Ornithodoros sp. cf. O. mimon). Eighty-one new associations between bird species and stages of tick species are detected. The families Thamnophilidae, Turdidae, Thraupidae, Passerellidae, Furnariidae and Troglodytidae were the most prevalent. According to the Phytogeographic Provinces involved in this study, the prevalence of infection for each of them in birds was: (1) Chaco: 28.2% (11 tick species); (2) Yungas: 22.0% (8 tick species); (3) Espinal: 11.1% (2 tick species); and (4) Pampa: 3.9% (1 tick species). This study provided information on the diversity of tick species that parasitize wild birds, the variability of the specific tick-bird associations between the different Phytogeographic Provinces and the relevance of some families of birds as hosts of different tick species.     

Molecular detection of a Borrelia sp. in nymphs of Amblyomma brasiliense ticks (Acari: Ixodidae) from Iguaçu National Park,Brazil, genetically related to Borrelia from Ethiopia and Côte d’Ivoire

Gram-negative spirochetes of the genus Borrelia are transmitted to vertebrate hosts through the tick bite during blood intake. Pathogenic Borrelia species may cause relapsing fever or Lyme borreliosis in humans. Our study aimed to molecularly detect and characterize bacteria of this genus in ticks collected in the Iguaçu National Park, located in southern Brazil. Ticks were collected from the environment (free living) and from hosts from May 2015 to July 2017. In total, 829 ticks were tested, being 741 from the environment and 88 from hosts; 128 ticks were larvae, 523 were nymphs, and 178 were adults (80 males and 98 females). The species identified were: Amblyomma brasiliense (42.9 %), Amblyomma coelebs (16.8 %), Amblyomma sp. (15.0 %), Amblyomma incisum (10.3 %), Amblyomma ovale (8.7 %), Haemaphysalis juxtakochi (5.5 %), Rhipicephalus microplus (0.5 %), Amblyomma longirostre (0.2 %), and Rhipicephalus sanguineus sensu lato (0.1 %). DNA extraction was performed with pools of larvae or nymphs, or individually with adults, for a total of 394 samples. The PCR technique used to detect Borrelia DNA was performed with two rounds of amplification reactions targeting a segment of the flagellin B gene (flaB). Amplification occurred in only one DNA sample, which was obtained from nymphs of A. brasiliense collected from the vegetation of a trail. The nucleotide sequence analysis revealed 90.8 % identity to a sequence of Borrelia sp. from Côte d’Ivoire, and 89.1 % identity to a sequence from Ethiopia. Phylogenetic analysis revealed that the Borrelia sequence from A. brasiliense was distinct from the Borrelia species of the groups belonging to Lyme borreliosis, relapsing fever and the one associated with reptile and echidna as hosts. The sequence is likely from a putative new species of Borrelia and was detected for the first time in A. brasiliense ticks.     

Morphological anomalies in hard ticks (Acari: Ixodidae) from Brazil

Tick abnormalities have been unusual in nature, and they can be divided into local and general. In the present study, external morphological anomalies were described in 31 individual adult ticks of 15 different species of Ixodidae, which were collected on wild hosts (20 ticks), domestic hosts (7 ticks), and in the environment (4 ticks) in 11 states of Brazil from 1998 to 2022. Among the 31 tick specimens, 14 (45%) were categorized as local anomalies, and 17 (55%) as general anomalies. The ticks were taxonomically identified into 14 species of Amblyomma, and one species of Rhipicephalus. Local anomalies included malformations of scutum/alloscutum, ectromely, leg atrophy, and a third ectopic spiracular plate. General anomalies included opisthosoma duplication, no expansion of dorsal alloscutum in engorged females, and gynandromorphism; the latter is described for 13 tick specimens. Morphological anomalies in Amblyomma aureolatum, Amblyomma brasiliense, Amblyomma humerale and Amblyomma longirostre are reported for the first time. Although the results herein expand the list of anomalous tick species in the Neotropics, future studies should be conducted to clarify the origin of these anomalies.     

Rickettsia parkeri Rickettsiosis, Argentina

Rickettsia parkeri, a recently identified cause of spotted fever rickettsiosis in the United States, has been found in Amblyomma triste ticks in several countries of South America, including Argentina, where it is believed to cause disease in humans. We describe the clinical and epidemiologic characteristics of 2 patients in Argentina with confirmed R. parkeri infection and 7 additional patients with suspected R. parkeri rickettsiosis identified at 1 hospital during 2004–2009. The frequency and character of clinical signs and symptoms among these 9 patients closely resembled those described for patients in the United States (presence of an inoculation eschar, maculopapular rash often associated with pustules or vesicles, infrequent gastrointestinal manifestations, and relatively benign clinical course). Many R. parkeri infections in South America are likely to be misdiagnosed as other infectious diseases, including Rocky Mountain spotted fever, dengue, or leptospirosis.Rickettsia parkeri, a tick-borne bacterium discovered in 1937, was considered nonpathogenic until 2004. Since 2004, >25 cases of R. parkeri rickettsiosis have been reported in persons living within the recognized range of the tick vector, Amblyomma maculatum, in the United States (1–4; Centers for Disease Control and Prevention, unpub. data). The clinical features of this newly recognized disease appear less severe than those produced by R. rickettsii bacteria, the agent of Rocky Mountain spotted fever (RMSF). For many years, investigators in several countries of South America, including Argentina, Brazil, and Uruguay, have recognized eschar-associated infections that clinically resemble R. parkeri rickettsiosis (5–7). These reports, and the discoveries of R. parkeri in A. triste ticks collected from these same countries, suggest that human infections with R. parkeri also occur in South America (8–10); to our knowledge, no confirmed cases of disease caused by this Rickettsia species have been reported from this continent.The Paraná Delta, situated in the provinces of Buenos Aires and Entre Ríos in Argentina, represents the terminus of the Paraná River as it approaches and drains into the Uruguay River and subsequently into the Río de la Plata. This alluvial ecosystem, where braided river branches create a network of islands and wetlands, covers ≈14,000 km² (5,405 mi²) and extends for ≈320 km (200 mi). This region also contains abundant populations of A. triste ticks (10). The Paraná Delta has always been a major agricultural and farming region. Recently, this area has become increasingly developed; roads have been built to allow greater access for tourism and recreational activities by many of the ≈14 million inhabitants of nearby Buenos Aires. In 2005, an eschar-associated febrile infection was diagnosed in a male beekeeper from the Paraná Delta; the infection was later confirmed as a spotted fever group (SFG) rickettsiosis by serology and immunohistochemistry (5). He had been bitten by a tick not far from several sites where R. parkeri was subsequently detected in A. triste ticks (10). We report confirmed cases of R. parkeri rickettsiosis in 2 patients in Argentina and describe additional suspected cases of this disease, or similar infections, in patients from the provinces of Buenos Aires, Chaco, and Entre Ríos.     

Novel PCR exclusion assay to detect spotted fever group rickettsiae in the lone star tick (Amblyomma americanum)

The lone star tick (Amblyomma americanum) is the most common and abundant human-biting tick in the southeastern United States where spotted fever rickettsioses frequently occur. However, the role of this tick in transmitting and maintaining pathogenic and non-pathogenic spotted fever group rickettsiae (SFGR) remains poorly defined. This is partially due to the high prevalence and abundance of Rickettsia amblyommatis in most populations of A. americanum. Many molecular assays commonly employed to detect rickettsiae use PCR primers that target highly conserved regions in the SFGR so low abundance rickettsia may not be detected when R. amblyommatis is present. It is costly and inefficient to test for low abundance rickettsial agents with multiple individual specific assays even when they are multiplexed, as most samples will be negative. Real time PCR assays may also be hampered by inadequate limits of detection (LODs) for low abundance agents. We exploited the absence of an otherwise relatively SFGR-conserved genome region in R. amblyommatis to design a hemi-nested PCR-assay which has a sensitivity of 10 copies in detecting the presence of most SFGR, but not R. amblyommatis in DNA of infected lone star ticks. This deletion is conserved in 21 isolates of R. amblyommatis obtained from multiple states. We demonstrated the assay’s utility by detecting a pathogenic SFGR, Rickettsia parkeri, in 15/50 (30 %) of field collected A. americanum ticks that were previously screened with conventional assays and found to be positive for R. amblyommatis. These co-infected ticks included 1 questing female, 6 questing nymphs, and 8 attached males. The high prevalence of R. parkeri among host-attached ticks may be due to several variables and does not necessarily reflect the risk of disease transmission from attached ticks to vertebrate hosts. This novel assay can provide accurate estimates of the prevalence of less common SFGR in A. americanum and thus improve our understanding of the role of this tick in the maintenance and transmission of the SFGR commonly responsible for human rickettsioses.