Ticks (Acari: Ixodidae) and rickettsiae associated with wild boars in a rural area of Minas Gerais, Brazil.

Wild boars or feral pigs are classified by the Brazilian Institute for the Environment and Renewable Resources (IBAMA) in "Category I of invasive exotic species". They cause economic losses, harm the environment, serve as hosts and reservoirs for several zoonotic disease agents, and provide a blood meal for tick species that act as vectors for zoonotic diseases. The objective of this study was to identify tick species on wild boars, assess host-seeking ticks in the related environment, and identify other potential tick hosts coexisting with wild boars on a farm located in the state of Minas Gerais, southeastern Brazil. Additionally, the study aimed to determine the presence of rickettsiae in these arthropods and assess the exposure of wild boars to rickettsiae species from the Spotted Fever Group and Rickettsia bellii through serology. A total of 3585 host-seeking ticks from three species (Amblyomma sculptum - 41.58%; Amblyomma dubitatum - 0.39% and Rhipicephalus microplus - 0.05%) were collected in the environment and A. sculptum was the most abundant species. Thirty-one wild boars were evaluated, resulting in the collection of 415 ticks, all of which were A. sculptum. Rickettsia DNA was not detected in samples of A. sculptum and R. microplus from the environment or in A. sculptum ticks from wild boars. However, all A. dubitatum ticks (n = 14) had Rickettsia bellii DNA confirmed by the species-specific PCR protocol. Out of the 31 serum samples from wild boars, 24 reacted with at least one Rickettsia antigen. Among these, seven individuals exhibited a reaction to a probable homologous antigen (PHA) of three rickettsiae species: R. rickettsii (n = 3), R. amblyommatis (n = 3) and R. rhipicephali (n = 1). Despite the high prevalence of seroreactivity, titers were low, indicating limited exposure to Rickettsia spp. Camera traps generated 874 animal records, capturing a total of 1688 individuals. At least 11 species of birds and 14 species of mammals (12 wild and two domestic) shared the environment with wild boars and potentially shared ticks with them. These findings provide baseline information for understanding the sharing of ticks and tick-borne pathogens between wild boars and other animals within the Cerrado biome. Further studies are necessary to monitor the potential and actual risk of wild boars to harbor infected ticks and their role in the transmission and maintenance cycle of Rickettsia spp.

Comparing scapular morphology of Amblyomma sculptum and Amblyomma dubitatum nymphs allows a fast and practical differential diagnosis of ticks in highly infested areas with dominance of these two species.

Rickettsia rickettsii infection causes the highest human fatality rate among all Rickettsia species of the world and is endemic in Southeast Brazil. In this part of the country most human spotted fever cases are related to unnaturally high populations of ticks, usually a mix of two species, Amblyomma dubitatum Neumann and Amblyomma sculptum Berlese and their local host, capybara (Hydrochoerus hydrochaeris). At the same time, an increase in the number of SF notifications as well as its urbanization was observed, and a better characterization of disease epidemiology is mandatory for control measures and to halt its expansion. It was recently noticed in southeast Brazil that SF endemic areas were characterized by overgrowth populations of A. sculptum and decreased populations of A. dubitatum. Hence, characterization of areas with potential to endemicity, eco-epidemiological studies and control measures may rely on the evaluation of A. sculptum/A. dubitatum rate. However, in potentially endemic scenarios, discriminating the two tick species, particularly the nymphs considered the main vector stage, is a challenge in face of several hundreds to thousands of ticks that should be examined for quantitative studies. We herein present additional morphological features to an existing identification key for Amblyomma nymphs that considerably diminishes the labor to distinguish nymphs of these two tick species. Specifically, the oval-shaped scutum of A. dubitatum and a conspicuous scapula of A. sculptum are prominent discriminating features.

Rhipicephalus microplus and Amblyomma sculptum (Ixodidae) infestation of Nellore cattle (Bos taurus indicus) in a farm of the Brazilian Cerrado: seasonality and infestation patterns.

We herein describe zebuine cattle tick infestation in a farm in southeast Brazil with an examination accurate enough to detect tick immatures and species other than R. microplus. Cattle were inspected monthly for ticks from May 2015 to May 2017 and 7604 ticks were collected along 276 bovine inspections. Rhipicephalus microplus was the dominant species (7197 specimens, 94.5% from the total), but Amblyomma sculptum was also collected (407/5.5%). Horse tick infestations were evaluated for comparison purposes of A. sculptum infestations of a primary host sharing pastures with bovines. Ticks were counted on the left side of 4-12 horses every 3 months from October 2015 to October 2017. Overall, 68 horse inspections were performed and 1702 ticks were collected: Dermacentor nitens (805 specimens/47.3% of the total), A. sculptum (733/43.1%) and R. microplus (164/9.6%). Overall mean tick abundance on bovines was low if compared to that of taurine cattle and counting revealed four annual generations of R. microplus. The interval between infestation peaks was 3 months, irrespective of the season, and an increase in tick counts from spring onward, as described in the south of Brazil, was not seen. Amblyomma sculptum infestation abundance was minor but constant and in high prevalence. Cattle infestation with A. sculptum seems to depend on pasture sharing with other domestic and wild hosts that are its primary hosts and provide engorged females to complete their life cycle. The impact of such tick sharing among several host species on tick-borne pathogen transmission remains to be elucidated.

Diversity of free-living ticks and serological evidence of spotted fever group Rickettsia and ticks associated to dogs, Porto Velho, Western Amazon, Brazil.

Rondônia is the only state in the North Region of Brazil to have registered confirmed cases of Brazilian Spotted Fever (BSF). The present study investigated the epidemiological cycle of Rickettsia spp. by surveying free-living ixodofauna and tick parasitism of dogs in the municipality of Porto Velho, Rondônia State. Ticks and dogs were tested for the presence of Rickettsia spp. DNA and dog serum was tested for reactivity to anti-Rickettsia spp. antibodies. Tick collection and dog blood sampling were performed in peri-urban and rural environments at 11 locations. Eight free-living Amblyomma species and one Haemaphysalis species were collected: A. scalpturatum, A. naponense, A. oblongoguttatum, A. coelebs, A. latepunctatum, A. pacae, A. ovale, Amblyomma sp., and H. juxtakochi. Three tick species were found parasitizing dogs: Rhipicephalus sanguineus sensu lato, A. oblongoguttatum and A. ovale. Molecular analysis did not identify the presence of the gltA gene fragment in any tick specimen. Results from an indirect immunofluorescent assay (IFA) showed that 20.8% of peri-urban and 15.4% of rural dog sera exhibited reactivity to Rickettsia rhipicephali, Rickettsia amblyommatis, Rickettsia bellii and Rickettsia parkeri antigens. Antibody prevalence in dogs was 16.4%. This study is the first to describe the prevalence of Rickettsia spp. infection in dogs from Porto Velho municipality. Our findings enhance current knowledge of Rickettsia spp. circulation in the Western Amazon.

Effect of low temperature and relative humidity on reproduction and survival of the tick Rhipicephalus microplus.

The objective of this study was to evaluate the survival of Rhipicephalus microplus females under low temperature, to determine the influence of temperature and relative humidity (RH) on the biological parameters of this tick. In total 300 viable engorged female ticks collected from cattle were used. On the day of collection (D0), 30 female ticks were divided into three groups: G1, kept at 28 °C and 80% RH; G2, kept at 20 °C and 80% RH; and G3, kept at 20 °C and 30% RH. The remaining females were stored in a refrigerator at 4 °C. Over the next 9 days (D+1 to D+9), 30 female ticks were removed daily from the refrigerator and distributed among the three treatments. Egg mass, feed conversion rate, egg incubation period, larval hatch rate, number of dead females per group, weight of female tick removed from the refrigerator, and female tick weight loss were recorded. At 20 °C and 80% RH hatching was delayed. Among the female ticks kept in the refrigerator and then allocated to groups G1, G2, and G3, there was a reduction in reproductive efficiency and a rise in mortality as time in the refrigerator increased. The female ticks expressed their best reproductive capacity when subjected to the refrigeration temperature for a maximum of 3 days and then kept at 28 °C and 80% RH. Thus, although female ticks can be stored in the refrigerator before being sent to research centers for use in bioassays, the storage time should be as short as possible, not exceeding 3 days.

Microhabitat determines uneven distribution of Amblyomma parvum but not of Amblyomma sculptum ticks within forest patches in the Brazilian Pantanal.

Environmental distribution of the two most abundant ticks in forest areas in the Brazilian Pantanal was evaluated by CO2 traps methodology in the wet season (peak of adult ticks) of 2012 and 2013. Adults of Amblyomma parvum were concentrated inside agglomerates of Bromelia balansae, in the border of forest patches. Adults of Amblyomma sculptum occurred in similar numbers both in bromeliad clumps and in bromeliad-free areas. Differential distribution of ticks in this habitat could be associated to the frequent use of bromeliad clumps by wild animals (potential hosts) and to the microclimate conditions inside this vegetation in the Pantanal. It is important to verify whether larvae and nymphs of A. parvum have a similar pattern of distribution in the same areas, during the dry season. These stages are more susceptible to desiccation and their principal hosts, non-volant small mammals, also use these bromeliad areas.

In vitro efficacy of Metarhizium anisopliae sensu lato against unfed Amblyomma parvum (Acari: Ixodidae).

Amblyomma parvum Aragão (Acari: Ixodidae) is a tick species found with wide distribution in the Neotropical region. Even though it is a wildlife-related tick, it is also a frequent parasite of domestic animals, is aggressive to human beings and may harbor pathogenic microorganisms. Therefore, it is a target species for control on domestic animals, particularly those at the rural-wildlife interface. Herein, the efficacy of two isolates (E9 and IBCB 425) of an entomopathogenic fungus, Metarhizium anisopliae sensu lato, already evaluated for ticks that parasitize domestic animals, was tested against unfed A. parvum adults. Both isolates displayed high acaricidal efficacy after immersion in fungal conidial suspensions for 5 min. Isolate E9 killed all ticks by the 7th day post-treatment, and isolate IBCB 425 did so by the 11th day. Tick mortality of 80 and 90% was achieved as early as the 3rd and 4th days, respectively, with both treatments. Thus, if a commercial M. anisopliae s.l. acaricide against domestic animal ticks is developed, it would also be effective against A. parvum.

Life cycle of Amblyomma mixtum (Acari: Ixodidae) parasitizing different hosts under laboratory conditions.

Amblyomma mixtum is a tick species in the Amblyomma cajennense complex. The known geographic range of A. mixtum extends from Texas in the USA to western Ecuador and some islands in the Caribbean. Amblyomma mixtum is a vector of disease agents of veterinary and public health importance. The objective of this study was to describe the life cycle of A. mixtum under laboratory conditions. Bovines, rabbits and sheep were infested with larvae, nymphs, and adult ticks under controlled conditions to assess several biological parameters. Eggs, larvae, nymphs and adults were kept in an incubator (27 °C temperature and 80% relative humidity) when they were off the host. The average life cycle of A. mixtum was 88 and 79 days when fed on rabbits and cattle, respectively. Sheep were found to be unsuitable because no ticks attached. The rabbit is a more practical host to maintain a colony of A. mixtum under laboratory conditions. The data from this study can be considered as an example for the life cycle of A. mixtum. However, caution must be exercised when making comparisons to the biology of A. mixtum in its natural habitat.