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.

Density and behavior of capybara (Hydrochoerus hydrochaeris) ticks (Acari: Ixodidae) Amblyomma sculptum and Amblyomma dubitatum with notes on Rickettsia bellii infection: Assessing human exposure risk.

In several urban and peri­urban areas of Brazil, populations of Amblyomma sculptum and Amblyomma dubitatum ticks are maintained by capybaras (Hydrochoerus hydrochaeris). In some of these areas, this host and these tick species are associated with Brazilian spotted fever (BSF), a lethal human disease caused by the bacterium Rickettsia rickettsii. In this work, we evaluated the risk of human exposure to these tick species using four collection techniques to discern host-seeking behavior. The study was carried out in 10 urban sites inhabited by capybaras in Uberlândia, a BSF-free municipality in southeastern Brazil. Ticks were collected in areas of 400 m2 at each site and at three seasons. Within the same municipality, the distance and speed of A. sculptum nymphs moving towards the CO2 traps were evaluated. In a sample of ticks Rickettsia DNA was investigated. During the study period, 52,953 ticks were collected. Among these, 83.4 % were A. sculptum (1,523 adults, 10,545 nymphs and 32,104 larvae) and 16.6 % were A. dubitatum (464 adults, 2,153 nymphs and 6,164 larvae). An average annual questing tick density of 4.4/m² was observed, with the highest density recorded at one site in autumn (31.8/m²) and the lowest in summer at another site (0.03/m²). The visual search yielded the highest proportion of A. sculptum larvae, constituting 47 % of the total and 63.6 % of all A. sculptum larvae. In contrast, CO2 traps collected a greater proportion of nymphs and adults of A. sculptum ticks. In the case of A. dubitatum, the CO2 trap was the most efficient technique with 57.7 % of captures of this species, especially of nymphs (94.5 % of captures) and adults (97.8 % of captures). Ticks' ambush height on vegetation (9 to 77 cm), observed by visual search 30 times, yielded a total of 20,771 ticks. Of these, 28 (93 %) were A. sculptum ticks, with only two (7 %) identified as A. dubitatum ticks. Among A. sculptum ticks, the nymph was the most attracted stage to humans and larva in the case of A. dubitatum. Amblyomma sculptum adults and nymphs were significantly more attracted to humans than those of A. dubitatum, but A. dubitatum larvae were significantly more attracted than the same stage of A. sculptum. The maximum distance and speed of horizontal displacement for A. sculptum nymphs were five meters and 2.0 m/h, respectively. The only species of Rickettsia detected in ticks, exclusively in A. dubitatum, was R. bellii. Importantly, it was observed that the higher the proportion of A. sculptum in the community of ticks, the lower the rate of infection of A. dubitatum by R. bellii. In conclusion, host-seeking behavior differed between the two tick species, as well as between stages of the same species. A greater restriction of A. dubitatum ticks to the soil was observed, while larvae and nymphs of A. sculptum dispersed higher in the vegetation. The behavior presented by A. sculptum provides greater opportunities for contact with the hosts, while A. dubitatum depends more on an active search for a host, the hunter behavior. Taken together, these observations show that a human being crossing an area infested with A. sculptum and A. dubitatum ticks will have almost exclusive contact with A. sculptum larvae and/or nymphs. Humans in a stationary position (sitting, lying or immobile) are exposed to both tick species, but they are more attractive to adults and mainly nymphs of A. sculptum compared to the corresponding stages of the tick A. dubitatum. The negative effect of A. sculptum on A. dubitatum infection by R. bellii deserves further studies.

Molecular detection of Borrelia sp. in Ornithodoros cavernicolous (Acari: Argasidae) in midwestern Brazil.

Ticks are obligate hematophagous parasites that can transmit to vertebrate hosts several pathogens, including viruses, bacteria, protozoa and helminths. Among these agents, some Borrelia species some Borrelia species cause disease in humans and other vertebrate hosts; therefore, they have medical and veterinary health importance. To gather additional information on Borrelia species in Brazil, the current study aimed to detect the presence of these species in Ornithodoros cavernicolous ticks collected in September 2019 from cement pipes that are used by bats as shelter in a farm located in the midwestern region of Brazil. DNA samples obtained from 18 specimens of O. cavernicolous were subjected of two polymerase chain reactions, targeting a segment of the Borrelia fla B gene. Of the samples tested, only one (6 %, 1/18) showed amplification. The nucleotide sequence of the amplified DNA showed more than 97 % (293/300) identity with a sequence of a Borrelia sp. detected in blood collected from a bat from Macaregua Cave, Colombia, and more than 97 % (292/300) detected in lungs from vampire bats from northeastern Brazil. The deduced amino acid sequences were identical to each other. Phylogenetic analysis indicated that these sequences formed a group of Borrelia species (putatively associated with bats) that is closely related to sequences of Borrelia species of the Lyme borreliosis group. Further investigations should be carried out in order to determine whether the sequence of the Borrelia sp. we found belongs to a new taxon. It will also be of great importance to determine which vertebrate hosts, besides bats, O. cavernicolous ticks can parasitize in order to investigate whether the Borrelia sp. we found may be transmitted and cause disease to the other vertebrate hosts.

Rickettsia communities and their relationship with tick species within and around the national park of Iguaçu, Brazil.

We report Rickettsia species from 2,334 ticks collected from environment (1,939 ticks) and animals (395 ticks) in the largest inland fragment of the Atlantic rainforest of southern Brazil and its fragments. Additionally, the DNA infection rates of Amblyomma ovale tick populations in the Neotropics with Rickettsia bellii and Rickettsia parkeri strain Atlantic rainforest were calculated using data from scientific publications, and their correlation was evaluated. From 11 tick species Rickettsia DNA was detected in seven (Amblyomma brasiliense, Amblyomma coelebs, Amblyomma incisum, Amblyomma longirostre, A. ovale, Haemaphysalis juxtakochi, Ixodes fuscipes) and was not detected in four species (Amblyomma dubitatum, Ixodes loricatus, Rhipicephalus microplus and Rhipicephalus sanguineus sensu lato). DNA of five Rickettsia species was detected (R. bellii, Rickettsia amblyommatis, Rickettsia rhipicephali, Rickettsia felis and Rickettsia sp. Aragaoi). To determine the prevalence of Rickettsia DNA positivity according to vector species, ticks were processed individually or in pools of 2-10 individuals (samples). The most prevalent Rickettsia species was R. bellii, found in 112 samples, followed by R. amblyommatis, R. rhipicephali, R. felis and Rickettsia sp. Aragaoi, found in 16, five, two and one sample, respectively. Rickettsia bellii DNA was found in five tick species with the highest infection rate in A. ovale and A. brasiliense. Absence of R. parkeri strain Atlantic rainforest in A. ovale ticks was an unexpected result. Furthermore, a negative correlation was identified between the infection rates (DNA) of R. bellii and/or R. parkeri strain Atlantic rainforest within A. ovale tick populations in the Neotropics. Putting together current knowledge, it can be proposed that, within natural settings, the diversity of rickettsiae and ticks creates a buffering effect on the overgrowth of rickettsiae and episodes of bacteremia in the hosts.

Occurrence of serological reactions for Leptospira spp. in donkeys and mules from Minas Gerais, Brazil.

Research concerning leptospirosis in donkeys and mules has been neglected around the world. Therefore, the aim of this study was to investigate the epidemiological situation of the prevalence of anti-Leptospira spp. antibodies in donkeys and mules from the state of Minas Gerais, Brazil. Blood serum samples were collected from 180 animals (109 donkeys and 71 mules) in two rural properties from the state of Minas Gerais, Brazil, and then submitted to a microscopic agglutination test (MAT). Urea and creatinine values were also quantified. Epidemiological variables such as age, breeding system, contact with other animal species, source of water and food, vaccination against leptospirosis, presence of reproductive alterations, and rodent control were also investigated. From 180 samples collected, 39 (21.67%) showed positive results in the MAT, at a dilution ≥ 1:100. Some animals were reactive for more than one serovar. The serovar Tarassovi was the most frequent (14.07%), followed by Hardjo (11.85%) and Wolffi (11.11%). There was a statistically significant difference between animals from 0 to 3 years of age reactive in the MAT in comparison to the other age groups. Most of the animals had urea and creatinine concentrations within the acceptable reference limit; however, there was a significant increase in creatinine levels in some of the test animals. The studied properties showed differences in some epidemiological aspects such as vaccination of the animals, presence of reproductive problems in the herd, and rodent control. Such aspects pointed as risk factors that may influence the frequency of positive serological results in property 1. The present study demonstrated that the prevalence of leptospirosis in donkeys and mules is high and several serovars are being maintained by these animals, representing a potential public health risk.

Ticks (Acari: Ixodidae) on marsh deer (Blastocerus dichotomus) at a conservation center: infestation and Rickettsia parkeri infection dynamics along nine years.

The objective of this work was to evaluate the relationship between the populations of the ticks Amblyomma triste Koch, 1844, Rhipicephalus microplus (Canestrini, 1888), Amblyomma sculptum Berlese, 1888, the pathogenic bacteria Rickettsia parkeri and a marsh deer (Blastocerus dichotomus) population after its removal from a pristine environment. For this purpose, ticks were collected from the cervical region of deer at the Marsh Deer Conservation Center in Promissão, São Paulo State, Brazil for nine consecutive years (2000 - 2008). Deer in captivity at the Center were kept in 2,000 m² paddocks surrounded by two-meter-high fences in the Tiete-river marsh. In total, 1,012 ticks of 26 deer were collected. Prevalence of the species A. triste among tick-infested hosts was the highest in the first triennium but decreased to the second and further to the third triennium. In contrast, the R. microplus prevalence amidst infested host population, increased from the first to the third triennium and was the species that attained the highest infestation intensity. Amblyomma sculptum was the tick with the lowest infestation prevalence and intensity throughout the period. The change in the proportion between the two most prevalent species was attributed to the new environment, specifically its restricted size and within it a dry area more suitable for R. microplus. DNA of 424 ticks processed in 276 pools was tested for Rickettsia genetic material by polymerase chain reactions (PCR). Twenty samples of the study were positive for the rickettsial gltA gene. Of these, 18 were from A. triste ticks and revealed the presence of the ompA spotted fever group gene as well. Eleven samples were sequenced and showed 100% identity with R. parkeri sensu stricto. Two samples from R. microplus did not amplify ompA gene neither yielded product in a PCR specific for Rickettsia bellii. Sequencing of the gltA gene in the DNA of these two ticks was also 100% identical with R. parkeri s.s. In conclusion, the changes in the deer environment modified the tick populations but maintained, at least temporary, R. parkeri bacteria in A. triste ticks. Rhipicephalus microplus was refractory to the R. parkeri infection and bacterial DNA in this tick species indicated DNA spill over from other tick species. It was demonstrated that captive marsh deer may sustain cattle tick populations on its own. This would hardly occur under pristine conditions because of the inadequacy the marsh deer´s naturally humid habitat for the cattle tick. However, deer transportations of R. microplus ticks from one farm to another may occur whenever habitat loss pushes this wild animal towards farms.

Free-living capybaras (Hydrochoerus hydrochaeris) in an urban area in Brazil: biochemical and hematological parameters

Capybaras have found favorable conditions for survival and reproduction in green urban environments. In recent years, the population of these large rodents has been increasingly abundant in several brazilian cities such as Uberlândia, a municipality of the southeastern region with a Cerrado biome. Capybaras are important in the Brazilian Spotted Fever epidemiological chain, by amplifying infection rates of the vector population. However, knowledge of this host's physiology is scarce. Thus, the aim of this work was to describe hematological and biochemical parameters of free-living capybaras groups in urbanized areas in the city of Uberlândia, Minas Gerais State, Brazil. Capybaras were captured in 4 different locations of Uberlândia city, Minas Gerais state, including 1 Condominium (P1), 1 Private Market Garden (P2), 1 Private Club (P3) and 1 Municipal Park (P4). The animals were baited into an octagonal iron corral and chemically contained with anesthetic darts. After sedated, blood was collected from the femoral vein in tubes with and without EDTA. Biochemical evaluation, hematological analysis with differential leukocyte counts and search for Dirofilaria sp. were done. The blood count and biochemistry values obtained from animals of different ages, sex and sectors (P1, P2, P3 and P4) were submitted to the Shapiro-Wilk normality test, considering 95% significance. Values that had a normal distribution were subjected to ANOVA tests followed by Student's t-test. Values that did not follow normality were submitted to the Kruskal-Wallis test, to obtain a P-value, with a significance level of 95%. A total of 19 capybaras were captured: 4 in P1, 6 in P2, 4 in P3 and 5 in P4. From the 19 animals, 13 were females (68.42%) and 6 were males (31.57%), 12 adults (63.15%) and 7 juveniles (36.84%). Apart from occasional skin scars and moderate to intense Ambyomma spp. tick infestations, all captured animals were healthy on a broad examination. From 5 animals captured in P4, despite the use of anticoagulant, blood from 4 animals clotted fast. No microfilariae were found in the thick drop test in any of the 19 animals sampled, and in 2 adult female capybaras captured in P1, Kurloff cells were observed. Hematological and biochemical values presented no major differences when comparing sex and age. Nevertheless, differences in liver and kidney profile were observed between the capybara groups, including ALT, alkaline phosphorus, BUN and creatinine. Blood from 4 animals clotted fast, despite the use of EDTA tubes. Blood clotting of samples with anticoagulant in this work could be associated with some physiological features inherent to capybaras. Many attempts were required to obtain enough blood from each individual due to the rapid hemostasis, what come in accordance with reports in literature. Kurloff cells were observed in 2 adult female capybaras captured in P1, which can be found in peripheral blood of female rodents during follicular phase of estrous cycle. Hematological and biochemical values differences in liver enzymes such as ALT and alkaline phosphorus, and kidney profile enzymes including BUN and creatinine could be associated to capture stress or dietetic variation between groups. Despite statistical relevant, the values were still in accordance with other works, although comparisons should be done with caution since various environments exert a diverse array of stimulus upon the animals such as parasitic, infective, stress, nutritional, social and undoubtedly blood parameters mirror them. In conclusion, this work contributes to the standardization of free-living capybaras' physiological parameters in urban areas.(AU)

Ticks and Rickettsia on anteaters from Southeast and Central-West Brazil.

The giant anteater (Myrmecophaga tridactyla) and the collared anteater (Tamandua tetradactyla) are widespread in Brazil and found in all Brazilian biomes. These hosts frequently use domestic animal environments such as pastures, where tick and related microorganism interchange may occur between hosts. Reports of tick infestations of these animals are scattered and refer to small samples and/or are geographically restricted. We herein present data on a wide geographic distribution of ticks and their Rickettsia collected from 72 giant and 30 collared anteaters, mostly road killed, over a period of 18 years, from Southeast and Central-West Brazil encompassing four States and 46 Municipalities. Overall nine tick species (Amblyomma auricularium, A. calcaratum, A. nodosum, A. ovale, A. parvum, A. sculptum, A. triste, Rhipicephalus microplus and R. sanguineus sensu lato) were collected from anteaters. Amblyomma sculptum, A. nodosum, and A. calcaratum were the most prevalent corresponding to, respectively, 48.8%, 39.3% and 2.7% of all ticks (n = 1775). However, A. nodosum tick numbers on collared anteaters were significantly higher (P < 0.001) than those on giant anteaters. At the same time, an abundance of A. sculptum adults on giant anteaters was significantly higher (Z = 2.875; P = 0.004) than that of A. nodosum and only eight A. sculptum nymphs were found on collared anteaters. DNA samples from 20 ticks from nine different animals yielded a visible amplicon in PCR targeting gltA. The PCR products targeting spotted-fever Rickettsia gene (ompA) from five adults of A. nodosum were sequenced and were shown to be 100% identical to Rickettsia parkeri strain NOD (MF737635.1). The product of one nymph and one adult of A. sculptum yielded a sequence 99% identical to R. parkeri strain NOD. Further, Rickettsia bellii genes were found in three A. nodosum adults. Ecological, behavioral and anatomical traits of anteaters are discussed to explain reported tick infestations and Rickettsia DNA found.

Effectiveness of monepantel and levamisole to control gastrointestinal strongylid parasites of sheep / Eficácia de monepantel e levamisol no controle de estrongilídeos parasitos gastrintestinais de ovinos

Anthelmintic resistance poses a major challenge to the control of endoparasites in sheep. The recent launch of monepantel may represent an alternative for the treatment of helminth infections. The purpose of this study was to assess the effectiveness of monepantel and levamisole in the sheep flock on the experimental farm Capim Branco, of the Federal University of Uberlandia, municipality of Uberlandia, state of Minas Gerais, Brazil. Thirty sheep with more than 200 eggs per gram of feces (EPG) were selected. Ten of these animals were treated orally with 2.5 mg/kg of monepantel, ten were treated subcutaneously with 7.5 mg/kg of levamisole, and ten served as control animals. Fecal samples for the determination of EPG were collected on days zero (pre-treatment), seven and fourteen (post-treatment). The percentage of effectiveness was calculated using RESO 2:01 software. Monepantel reduced the EPG of Haemonchus sp. and Trichostrongylus sp. by more than 95%, but Oesophagostomum sp. was reduced by only 50% and 33% on days seven and fourteen. The percentage reduction of EPG by levamisole remained above 95% for all the helminths recovered from fecal cultures, indicating its effectiveness. These data demonstrate the failure of monepantel to control Oesophagostomum spp. in the evaluated sheep flock.

A resistência anti-helmíntica é um dos maiores desafios para o controle de endoparasitos em ovinos. O recente lançamento do monepantel pode representar uma alternativa para o tratamento das helmintoses. Este trabalho objetivou avaliar a eficácia do monepantel e levamisol no rebanho ovino da Fazenda Capim Branco da Universidade Federal de Uberlândia, em Uberlândia, Minas Gerais. Foram selecionados trinta ovinos com valores de ovos por grama de fezes (OPG) superiores a 200. Destes animais, dez foram tratados com monepantel 2,5 mg/kg, via oral, dez receberam tratamento subcutâneo com levamisol 7,5 mg/kg, e os outros 10 permaneceram como animais controle. No dia zero (prétratamento), sete e quatorze (pós-tratamento), realizou-se coletas de fezes para determinação de OPG. O percentual de eficácia foi calculado pelo software RESO 2.01. O monepantel apresentou percentual de redução superior a 95% para Haemonchus sp. e Trichostrongylus sp., porém este percentual para Oesophagostomum sp. foi de 50% e 33%, no sétimo e décimo quarto dia. O percentual de redução de OPG para levamisol se manteve superior a 95% para todos os helmintos recuperados nas coproculturas, indicando sua eficácia. Estes dados indicam a falência do monepantel em realizar o controle de Oesophagostomum spp. no rebanho avaliado.