Invasive honeysuckle eradication reduces tick-borne disease risk by altering host dynamics.

Despite the ubiquity of invasive organisms and their often deleterious effects on native flora and fauna, the consequences of biological invasions for human health and the ecological mechanisms through which they occur are rarely considered. Here we demonstrate that a widespread invasive shrub in North America, Amur honeysuckle (Lonicera maackii), increases human risk of exposure to ehrlichiosis, an emerging infectious disease caused by bacterial pathogens transmitted by the lone star tick (Amblyomma americanum). Using large-scale observational surveys in natural areas across the St. Louis, Missouri region, we found that white-tailed deer (Odocoileus virginianus), a preeminent tick host and pathogen reservoir, more frequently used areas invaded by honeysuckle. This habitat preference translated into considerably greater numbers of ticks infected with pathogens in honeysuckle-invaded areas relative to adjacent honeysuckle-uninvaded areas. We confirmed this biotic mechanism using an experimental removal of honeysuckle, which caused a decrease in deer activity and infected tick numbers, as well as a proportional shift in the blood meals of ticks away from deer. We conclude that disease risk is likely to be reduced when honeysuckle is eradicated, and suggest that management of biological invasions may help ameliorate the burden of vector-borne diseases on human health.

Tunga penetrans and further parasites in the giant anteater (Myrmecophaga tridactyla) from Minas Gerais, Brazil.

The Brazilian savannah-like area, the Cerrado region, covers large areas of the country and provides a habitat for a multitude of different animal species. The giant anteater (Myrmecophaga tridactyla) is geographically widespread and one of the typical inhabitants of the Cerrado. They are listed as vulnerable by the International Union for Conservation of Nature and Natural Resources. A population loss of at least 30 % over the past 10 years has been estimated based on local extinctions, habitat loss and deaths caused by fires, roadkills and hunting. Little is known about ecological and in particular parasitic conditions of this highly specialised insectivore species. During September and November 2010 we examined three roadkilled giant anteater for the presence of metazoan ecto- and endoparasites. Besides the cestode species Oochoristica tetragonocephala and the tick species Amblyomma nodosum, we found for the first time the flea Tunga penetrans. Beside morphological flea species identification, we compared a fragment of the 16S rRNA gene of eggs, a molecular method utilised for the first time in this combination. The identification of T. penetrans in M. tridactyla represents a new host record and expands the host and distribution range of the zoonotic flea species.

Sudden Mortality in Captive White-Tailed Deer With Atypical Infestation of Winter Tick.

In October 2020, three captive male white-tailed deer, Odocoileus virginianus [Zimmermann] (artiodactyla: Cervidae), were found dead in central Pennsylvania and a fourth was euthanized due to extreme lethargy. The deer presented with high burdens of Dermacentor albipictus (Packard) (Ixoda: Ixodidae) (winter tick). There were no other clinical symptoms and deer were in otherwise good physical condition with no observed alopecia. Winter tick epizootics have been associated with mortalities of moose, Alces alces [Linnaeus] (artiodactyla: cervidae), and more recently elk, Cervus canadensis [Erxleben] (artiodactyla: cervidae), in Pennsylvania, but have not been reported in white-tailed deer. Mild winters are favorable to winter ticks and deer producers and managers should be aware of possible infestations as a result.

Investigation of Ixodid ticks as vectors of Babesia caballi and Theileria equi (Protozoa: Apicomplexa) in central Italy.

Babesia caballi and Theileria equi are widely recognized as causative agents of equine pirolasmosis (EP), an acute, sub-acute, and chronic disease of equines, with relevant economic impact on horse trade worldwide. Although several studies on EP prevalence from central Italy have been published, data on ticks responsible for its transmission are still lacking. In this study, we identified a potential competent vector, investigating main features of its ecology together with EP infection rates. A two-year sampling of questing ticks was carried out for the first time in Italy in an area known for high EP prevalence in horse sera, detecting the association between Rhipicephalus bursa and causative agents of EP. Most of the positive pools harbored a single infection (91.1%); mixed infections were also detected (8.9%). The infection rate for T. equi slightly decreased among years; B. caballi showed a lower, but increasing, infection rate. Tick phenology, climate variables, and peaks of EP prevalence indicated late May and second half of June as periods with the highest risk of new infections, especially during warm and dry days.

Amblyomma americanum serpin 27 (AAS27) is a tick salivary anti-inflammatory protein secreted into the host during feeding.

Ticks successfully feed and transmit pathogens by injecting pharmacological compounds in saliva to thwart host defenses. We have previously used LC-MS/MS to identify proteins that are present in saliva of unfed Amblyomma americanum ticks that were exposed to different hosts. Here we show that A. americanum serine protease inhibitor (serpin) 27 (AAS27) is an immunogenic saliva protein that is injected into the host within the first day of tick feeding and is an anti-inflammatory protein that might act by blocking plasmin and trypsin functions. Although AAS27 is injected into the host throughout tick feeding, qRT-PCR and western blotting analyses indicate that the respective transcript and protein are present in high amounts within the first 24 h of tick feeding. Biochemical screening of Pichia pastoris-expressed recombinant (r) AAS27 against mammalian proteases related to host defense shows it is an inhibitor of trypsin and plasmin, with stoichiometry of inhibition indices of 3.5 and 3.8, respectively. Consistent with typical inhibitory serpins, rAAS27 formed heat- and SDS-stable irreversible complexes with both proteases. We further demonstrate that rAAS27 inhibits trypsin with ka of 6.46 ± 1.24 x 104 M-1 s-1, comparable to serpins of other tick species. We show that native AAS27 is part of the repertoire of proteins responsible for the inhibitory activity against trypsin in crude tick saliva. AAS27 is likely utilized by the tick to evade the hosts inflammation defense since rAAS27 blocks both formalin and compound 48/80-induced inflammation in rats. Tick immune sera of rabbits that had acquired resistance against tick feeding following repeated infestations with A. americanum or Ixodes scapularis ticks reacts with rAAS27. Of significant interest, antibody to rAAS27 blocks this serpin inhibitory functions. Taken together, we conclude that AAS27 is an anti-inflammatory protein secreted into the host during feeding and may represent a potential candidate for development of an anti-tick vaccine.

Chemical Composition and Acaricidal Activity of the Essential Oils of Some Plant Species of Lamiaceae and Myrtaceae against the Vector of Tropical Bovine Theileriosis: Hyalomma scupense (syn. Hyalomma detritum).

The present study aimed to investigate the acaricidal properties of six essential oils. They were extracted from some plant species (Lamiaceae and Myrtaceae) using the technique of hydrodistillation with the Clevenger apparatus. The chemical compositions of the essential oils under study were determined by gas chromatography-mass spectrometer (GC-MS). An Adult Immersion Test (AIT) and a Larval Immersion Test (LIT) were used to evaluate the acaricidal activity of these essential oils against the adults and larvae of Hyalomma scupense. GC-MS analysis showed the major constituents of each essential oil: 25.49% of α-thujone (lavender); 46.82% of carvacrol (oregano); 78.78% of carvacrol (thyme); 40.27% of 1,8-cineole (blue gum); 17.45% of p-cymene (river red gum); and 26.96% of 1,8-cineole (rosemary). The biotests on the essential oils revealed that they inhibit the reproduction of H. scupense engorged females at a rate of 100 % with doses of 0.781 µl/ml of rosemary, 1.562 µl/ml of thyme, 3.125 µl/ml of lavender and oregano, and 6.250 µl/ml of blue gum and river red gum. After a treatment that lasted for 24 hours, essential oils showed a larvicidal activity with respective values of lethal concentrations (LC): LC50, LC90, and LC95 (0.058, 0.358, and 0.600 µl/ml for thyme; 0.108, 0.495, and 0.761 µl/ml for rosemary; 0.131, 0.982, and 1.740 µl/ml for oregano; 0.155, 2.387, and 5.183 µl/ml for blue gum; 0.207, 1.653, and 2.978 µl/ml for river red gum; and 0.253, 2.212, and 4.092 µl/ml for lavender). This is the first report on the acaricidal activity of these essential oils against H. scupense. The results obtained showed that the essential oils with chemotype carvacrol, 1,8-cineole, α-thujone, and p-cymene are highly acaricidal, and they can be used for ticks control. However, further studies on their toxicity in nontarget organisms are required.

Transstadial transmission of Borrelia turcica in Hyalomma aegyptium ticks.

Borrelia turcica comprises the third major group of arthropod-transmitted borreliae and is phylogenetically divergent from other Borrelia groups. The novel group of Borrelia was initially isolated from Hyalomma aegyptium ticks in Turkey and it was recently found in blood and multiple organs of tortoises exported from Jordan to Japan. However, the ecology of these spirochetes and their development in ticks or the vertebrate hosts were not investigated in detail; our aims were to isolate the pathogen and to evaluate the possibility of transstadial transmission of Borrelia turcica by H. aegyptium ticks. Ticks were collected from Testudo graeca tortoises during the summer of 2013 from southeastern Romania. Engorged nymphs were successfully molted to the adult stage. Alive B. turcica was isolated from molted ticks by using Barbour-Stoenner-Kelly (BSK) II medium. Four pure cultures of spirochetes were obtained and analyzed by PCR and sequencing. Sequence analysis of glpQ, gyrB and flaB revealed 98%-100% similarities with B. turcica. H. aegyptium ticks collected from T. graeca tortoises were able to pass the infection with B. turcica via transstadial route, suggesting its vectorial capacity.

Notes on parasitism by Amblyomma humerale (Acari: Ixodidae) in the state of Rondônia, western Amazon, Brazil.

The tick Amblyomma humerale Koch is endemic to South America. All host records refer to the adult stage parasitizing tortoises, mostly yellow-footed tortoise, Geochelone denticulata (L.), and red-footed tortoise, Geochelone carbonaria (Spix). The current study reports the presence of A. humerale in the state of Rondônia, Brazil. A total of 215 adult ticks (201 males, 14 females) was collected from six G denticulata in an Indian reserve and nine Geochelone sp. in rural Monte Negro County, giving an overall mean infestation of 14.3 +/- 12.0 (range: 2-44) ticks per tortoise. Male ticks always outnumbered females on the host and nine tortoises had only male ticks. Male ticks were mostly attached in clusters on the ventral sides of the carapace near the anterior and posterior margins, and more rarely on the outer margin of the plastron. All females were found attached to the tortoise skin, at different sites such as head, neck, shoulders or legs. Male ticks were rarely observed attached to the body skin. Seven engorged nymphs collected on small vertebrates from Monte Negro County molted to adults of A. humerale. This included one nymph each on the seven-colored lizard, Plica plica (L), green tree climber, Plica umbra (L.), and wide-foraging lizard, Kentropyx calcarata Spix,three nymphs on the common opossum, Didelphis marsupialis L., and one nymph on the silky anteater, Cyclopes didactylus L. These constitute the first host records for the immature stages of the tick A. humerale.

Study of the seasonal dynamics, life cycle, and host specificity of Amblyomma aureolatum (Acari: Ixodidae).

In the first part of this study, monthly infestation by ticks was evaluated on dogs from December 2000 to November 2002 in the rural area of Taiaçupeba, São Paulo. Adults of Amblyomma aureolatum (Pallas) were found on dogs in all months, with a mean prevalence per month of 46.9 +/- 15.7% (range, 25-80%). The mean tick relative abundance per month was 2.4 +/- 2.7 ticks (range, 0.5-14 ticks), and the mean tick mean intensity per month was 4.7 +/- 4.2 ticks (range, 1.5-23.3 ticks). No A. aureolatum immature ticks were found on dogs. In the second part of this study, we studied the life cycle of A. aureolatum in the laboratory. We tested the suitability of six host species for the immature stages and dogs for the adult stage. Tick developmental periods were observed at different temperatures (23, 25, or 27 degrees C), always with RH >95%, which were satisfactory for all free-living developmental stages of the tick life cycle. Chickens and guinea pigs were the most suitable hosts for larvae and nymphs (recovery rates, 18.4-52.2%). Dogs were highly suitable for adult ticks (all females exposed to them were recovered and laid eggs) but were unsuitable for the immature ticks (recovery rates, 0-10%). Based on published host records for A. aureolatum, our results indicate that dogs and birds are primary hosts for adult and immature stages, respectively, of A. aureolatum in nature. In addition, wild guinea pigs are indicated as another potential primary host for immature ticks.

Pathogenicity of entomopathogenic fungi Beauveria bassiana and Metarhizium anisopliae to Ixodidae tick species Dermacentor variabilis, Rhipicephalus sanguineus, and Ixodes scapularis.

Nymphal and adult ticks from three different tick species, Dermacentor variabilis Say, Ixodes scapularis Say, and Rhipicephalus sanguineus Latrielle, were treated with conidia and blastospores of the entomopathogenic fungi Beauveria bassiana (Bals.) Vuill. and Metarhizium anisopliae Metschnikoff. Dose-response experiments indicated that a critical concentration of fungal spores is required for infection and mortality. Over a 28-d time course, fungal suspensions of either B. bassiana or M. anisopliae at 10(8) conidia/ml resulted in 50-70% mortality in adult I. scapularis and R. sanguineus, but <20% mortality in D. variabilis ticks. R. sanguineus nymphs were highly susceptible to both entomopathogenic fungi, displaying >60% mortality within 14 d postinfection and >90% mortality within 21-28 d postinfection. D. variabilis nymphs also were more susceptible than their corresponding adults, displaying mortalities ranging from 20 to 40% 28 d postinfection. I. scapularis nymphs, however, seemed to be slightly less susceptible than adults (45% mortality, 28 d postinfection). The addition of nutrients to fungal cell suspensions did not have any noticeable effects on mortality toward any of the tick species tested. Significant mortality against D. variabilis adults (approximately 65%) was noted only when B. bassiana fungal cells with growth media carryover were used as the inoculum against the ticks. Entomopathogenic fungi such as B. bassiana and M. anisopliae may have the potential for controlling populations of I. scapularis and R. sanguineus, and under certain conditions D. variabilis. Our results indicate that inoculum conditions can greatly affect successful virulence and subsequent mortality.

[Dichotomous identification keys of ticks (Acari: Ixodidae), livestock parasites in North Africa]. / Clé dichotomique et identification des tiques (Acari: Ixodidae) parasites du bétail au Maghreb.

The known fauna of livestock in the Maghreb consists of 5 established species. Five other species recorded seem to be introduced by imported animals. In this paper, dichotomous keys are given for species and subspecies of the genus Hyalomma. Rhipicephalus and Haemaphysalis parasite of livestock in North Africa. For the ticks of the other genera which are represented by only one species, we provide an identification description. Notes on geographical distribution, host and period of activity are provided.

Seasonal and spatial distribution of ixodid tick species feeding on naturally infested dogs from Eastern Austria and the influence of acaricides/repellents on these parameters.

BACKGROUND: Effective control of tick infestation and pathogen transmission requires profound knowledge of tick biology in view of their vector function. The particular time of the year when the different tick species start to quest and the favoured sites on the canine host are of major interest. The efficacy of acaricides/repellents to control ticks in the field requires observation. METHODS: To address these issues, 90 dogs, grouped in "untreated", "acaricide/repellent" (permethrin) and "acaricide only" (fipronil) animals and subjected to tick infestation under natural conditions in Burgenland (Eastern Austria), were examined. The number and species of ticks occurring during and outside the protection time was evaluated during a period of 11 months and the biting location on the dogs' skin was recorded. RESULTS: Of the 700 ticks collected, the most common species in that particular walking area was Ixodes ricinus, followed by Dermacentor reticulatus and Haemaphysalis concinna. Regarding the on-host activity, D. reticulatus displayed more infestations in early spring and late autumn, whereas I. ricinus occurred almost one month later in spring and one month earlier in autumn. H. concinna followed a monophasic pattern of activity with a peak in summer. The preferred feeding sites of the ticks on the dogs were on the head, neck, shoulder and chest. This distribution over the dog's body was not influenced by the use of the drugs, although on the whole fewer ticks (22.5% of all ticks) were found during the protection time. Interestingly, differences occurred with the use of drugs compared to non-protected dogs with regard to the infestation over the year. Acaricide-treated dogs displayed a higher prevalence in April, May and September, whereas dogs of the acaricide/repellent group showed a higher infestation in March, July, October and November. CONCLUSION: The different tick species display different on-dog activity peaks over the year, during which particular canine diseases can be expected and predicted, considering the specific incubation times for each pathogen.The tick species occurring in this study do not seem to choose particular sites on the dogs. Their arrival place seems to represent the attachment and consequently the feeding sites. The use of acaricides leads to a significantly (p<0.01) lower number of infesting ticks but no change of the distribution pattern on the dogs was observed.

Primeiro registro de machos de Amblyomma rotundatum (Acari: Ixodidae) em jabuti-tinga (Chelonoidis denticulatus) no estado do Amazonas, Amazônia brasileira: relato de caso / First record of males of Amblyomma rotundatum (Acari: ixodidae) on yellow-footed tortoise (Chelonoidis denticulatus) in the Amazonas state, Brazilian Amazon: case report

O carrapato Amblyomma rotundatum é uma espécie partenogenética, encontrada principalmente em anfíbios e répteis silvestres em todos os biomas brasileiros. No presente relato, registra-se a ocorrência de dois machos dessa espécie de ixodídeo encontrados em um jabuti-tinga (Chelonoidis denticulatus) proveniente do município de Manaus, Amazonas, Brasil.(AU)

The Amblyomma rotundatum tick is a parthenogenetic species, found mainly in wild amphibians and reptiles in all Brazilian biomes. The present report records the occurrence of two males of this ixodid species found on yellow-footed tortoise (Chelonoidis denticulatus) from the municipality of Manaus, Amazonas, Brazil.(AU)

Importation of Hyalomma marginatum, vector of Crimean-Congo haemorrhagic fever virus, into the United Kingdom by migratory birds.

Hyalomma marginatum ticks are an important vector of Crimean-Congo haemorrhagic fever virus which can result in a severe and potentially fatal disease in humans. Given the continued emergence of clinical cases in Eurasia and focalised upsurges of H. marginatum populations in Europe, it seemed prudent to assess the potential of this vector species to be introduced into the United Kingdom. Immature forms of H. marginatum are frequent ectoparasites of passerine birds many of which migrate from Africa to the UK each spring. Incoming birds were inspected for ticks during the spring migration in 2010 and 2011. A total of 68 ticks was collected from 971 birds (29 bird species), 21% (14) of the ticks were identified as H. marginatum. Oenanthe oenanthe (Northern wheatear) and Sylvia communis (Whitethroat) were found to be infested by this tick in both years and with multiple ticks. Single specimens were also removed from Acrocephalus schoenobaenus (Sedge warbler) and Phoenicurus phoenicurus (Common redstart) in 2010. This study provides the first contemporary evidence for substantial importation of this tick species into the UK.

Efficacy of Hyalomma scupense (Hd86) antigen against Hyalomma excavatum and H. scupense tick infestations in cattle.

The Rhipicephalus microplus recombinant Bm86-based tick vaccines have shown their efficacy for the control of several Hyalomma cattle ticks genera, namely H. dromedarii and H. anatolicum. However, H. scupense species, the most important tick in North Africa has never been studied. Vaccination trials using either a recombinant Bm86-based vaccine or a recombinant Hd86-based vaccine (the Bm86 ortholog in H. scupense) were conducted in cattle against immature and adult H. scupense ticks and adult H. excavatum ticks. The results showed a 59.19% reduction in the number of scupense nymphs engorging on Hd86 vaccinated cattle. However, cattle vaccination with Bm86 or Hd86 did not have an effect on H. scupense or H. excavatum adult ticks infestations. These results showed that Hd86 vaccines are selectively effective against H. scupense immature instars and emphasize on an integrated anti-tick vaccine control in North Africa.

Efficacy of Rhipicephalus (Boophilus) microplus Bm86 against Hyalomma dromedarii and Amblyomma cajennense tick infestations in camels and cattle.

The recombinant Bm86-based tick vaccines have shown their efficacy for the control of cattle ticks, Rhipicephalus (Boophilus) microplus and R. annulatus infestations. However, cattle ticks often co-exist with multi-host ticks such as Hyalomma and Amblyomma species, thus requiring the control of multiple tick infestations for cattle and other hosts. Vaccination trials using a R. microplus recombinant Bm86-based vaccine were conducted in cattle and camels against Hyalomma dromedarii and in cattle against Amblyomma cajennense immature and adult ticks. The results showed an 89% reduction in the number of H. dromedarii nymphs engorging on vaccinated cattle, and a further 32% reduction in the weight of the surviving adult ticks. In vaccinated camels, a reduction of 27% and 31% of tick engorgement and egg mass weight, respectively was shown, while egg hatching was reduced by 39%. However, cattle vaccination with Bm86 did not have an effect on A. cajennense tick infestations. These results showed that Bm86 vaccines are effective against R. microplus and other tick species but improved vaccines containing new antigens are required to control multiple tick infestations.

Longistatin is an unconventional serine protease and induces protective immunity against tick infestation.

Classical serine proteases use the conserved Ser/His/Asp catalytic triad to hydrolyze substrates. Here, we show that longistatin, a salivary gland protein with two EF-hand domains from the vector tick Haemaphysalis longicornis, does not have the conserved catalytic triad, but still functions as a serine protease. Longistatin was synthesized in and secreted from the salivary glands of ticks, and is injected into host tissues during the acquisition of blood-meals. Longistatin hydrolyzed fibrinogen, an essential plasma protein in the coagulation cascade, and activated plasminogen, into its active form plasmin, a serine protease that dissolves fibrin clots. Longistatin efficiently hydrolyzed several serine protease-specific substrates showing its specificity to the amide bond of Arg. Longistatin did not hydrolyze synthetic substrates specific for other groups of proteases. The enzyme was active at a wide range of temperatures and pHs, with the optimum at 37°C and pH 7. Its activity was efficiently inhibited by various serine protease inhibitors such as phenylmethanesulfonyl fluoride (PMSF), aprotinin, antipain, and leupeptin with the estimated IC(50) of 278.57 µM, 0.35 µM, 41.56 µM and 198.86 µM, respectively. In addition, longistatin was also potently inhibited by Zinc (Zn(2+)) in a concentration-dependent manner with an IC(50) value of 275 µM, and the inhibitory effect of Zn(2+) was revived by ethylenediaminetetra acetic acid (EDTA). Immunization studies revealed that longistatin sharply induced high levels of protective IgG antibodies against ticks. Immunization with longistatin reduced repletion of ticks by about 54%, post engorgement body weight by >11% and molting of nymphs by approximately 34%; thus, the vaccination trial was approximately 73% effective against tick infestation. Taken together, our results suggest that longistatin is a new potent atypical serine protease, and may be an interesting candidate for the development of anti-tick vaccines.