Permethrin-induced ultrastructural changes in oocytes of Rhipicephalus sanguineus (Latreille, 1806) (Acari: Ixodidae) semi-engorged females

Currently, the most effective method for tick control is by the use of chemical acaricides such as permethrin (active ingredient of the Advantage^® Max3 acaricide, Bayer), a pyrethroid with neurotoxic action. However, data about its effect on other tick systems, such as glandular and reproductive, are still scarce. This study provides ultrastructural details of the toxic effects induced in oocytes of semi-engorged Rhipicephalus sanguineus females exposed to 206, 1031, and 2062 ppm of permethrin. The results showed that permethrin is an effective acaricide by inhibiting and/or neutralizing the tick reproductive process since oocytes of treated ticks presented major structural changes such as cytoplasm vacuolation, disruption of both membranes and cell organelles, leading to cell death. Data also showed that even in low concentrations permethrin induced changes in germ cells, making them non-viable. The defense mechanisms used by oocytes against the action of permethrin in the attempt of recovering the cellular integrity and of ensuring the viability of these cells were also described in this study.     

Status of ivermectin resistance in Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) populations from north-western Himalayas,India

Status of ivermectin resistance was studied in 16 isolates of Rhipicephalus (Boophilus) microplus ticks collected from north-western Himalayan region of India using larval immersion test (LIT). Regression graphs of probit mortality of larval ticks plotted against log values of concentrations of ivermectin were utilized for the determination of slope of mortality, lethal concentrations (LC₅₀ and LC₉₅) and resistance ratios (RRs). Out of 16 isolates examined, 15 were found resistant against ivermectin. Level II resistance was detected in eleven isolates (RR₅₀ ranged from 5.72 to 7.62), whereas four isolates exhibited level I resistance (RR₅₀ ranged from 1.58 to 2.77). Only one isolate was found susceptible (RR₅₀: 1.23). Higher RR values were observed in tick isolates from low altitude areas where intensive animal husbandry activities are practiced as compared to tick isolates of high altitudes accredit the fact that managemental practices, availability of drug and its frequency of application play an important role in development of resistance.     

Action of permethrin on Rhipicephalus sanguineus (Latreille, 1806) (Acari: Ixodidae) semi-engorged females: Morpho-physiological evaluation of salivary glands

Currently the most effective method of tick control is the use of acaricides, among which stands out permethrin (active ingredient of acaricide Advantage^® Max3, Bayer), a neurotoxic pyrethroid. However, assessments of their effects on other tick systems such as glandular are still scarce. Thus, this study provides information, through histochemical techniques, about the toxic effect of this pyrethroid on the morphophysiology of salivary glands of semi-engorged Rhipicephalus sanguineus females exposed to different concentrations of permethrin (206, 1031, and 2062 ppm). The results showed that permethrin caused significant changes in the salivary gland metabolism accelerating the process of glandular degeneration, an event which would occur naturally and with great intensity only in the final engorgement stage. Furthermore, this study pointed out that permethrin reduces the salivary gland secretion ability through a drastic reduction of proteins, lipids, and polysaccharides in acinar cells. These changes impair the females to finalize the feeding process, what indirectly affects the reproductive process.     

Effects of carvacrol and thymol on the antioxidant and detoxifying enzymes of Rhipicephalus microplus (Acari: Ixodidae)

The present study was conducted to evaluate the effects of carvacrol and thymol on the antioxidant and detoxifying enzymes of larvae from two populations of R. microplus: Jaguar (tick population resistant to six classes of acaricides) and Porto Alegre (susceptible tick population). Carvacrol and thymol were tested at concentrations ranging from 0.14 to 5.0 mg mL^?1 in both populations to determine the LC₅₀. In addition, the LC₁, LC₂₅, and LC₇₅ were estimated using the LC₅₀ and HillSlope of each compound. Larvae of both populations of R. microplus were then treated with the LC₁, LC₂₅, LC₅₀, and LC₇₅ of each monoterpene, and those that survived were processed to evaluate the effects of the compounds on the antioxidant and detoxifying systems of larvae; these effects were assessed by determining the activity of the enzymes, glutathione-S-transferase (GST), catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GPX). Larvae from the Jaguar population treated with different lethal concentrations of carvacrol and thymol displayed a dose-dependent increase in CAT, GPX, SOD, and GST after treatment with LC₂₅. Further, larvae treated with the LC₇₅ had the highest levels of enzyme activity for carvacrol (1.76 mg mL^?1) and thymol (1.32 mg mL^?1). CAT, GPX, SOD, and GST activity in Porto Alegre population larvae treated with carvacrol and thymol also increased significantly up to the LC₅₀ of each monoterpene. However, at the LC₇₅ of carvacrol and thymol, a decrease in the activity of all enzymes was observed for this tick population. These findings indicate that carvacrol and thymol induced increased activity of all evaluated enzymes at different lethal concentrations in R. microplus larvae from two populations. Such findings unveil the possible mechanisms of action of these natural acaricides.     

Thymol: Effects on reproductive biology and Gene's organ morphology in Rhipicephalus sanguineus sensu lato engorged females (Acari: Ixodidae)

Thymol is a monoterpene with proven acaricidal activity on different tick species and life stages. The objective of this work was to evaluate the effect of thymol on the reproductive biology of engorged females of Rhipicephalus sanguineus sensu lato and the morphological changes caused in the cells of the tubular and accessory glands of Gené’s organ. The females were exposed to thymol by immersion. Seven groups were established: group I (distilled water), group II (30% ethanol), group III (thymol 1.25 mg/mL), group IV (thymol 2.5 mg/mL), group V (thymol 5.0 mg/mL), group VI (thymol 10.0 mg/mL), and group VII (thymol 20.0 mg/mL), with 20 replicates for each treatment. The ticks were kept in a BOD incubator at 27 ± 1 °C and RH > 80 ± 10%. Ten females from each group were evaluated daily until death, and the remaining ten were kept in the incubator under the same conditions for five days and then dissected for the removal of Gené’s organ to note possible damage to cell morphology by scanning electron microscopy (SEM) and hematoxylin-eosin (HE) techniques. Thymol demonstrated 63% and 98% efficacy in groups VI (10.0 mg/mL) and VII (20.0 mg/mL), respectively. In these groups, the cells of the tubular and accessory glands of Gené’s organ showed signs of damage: irregular eosin staining, rupture and deformation of the cellular limit, presence of fragmented nuclei, changes in cytoplasmic homogeneity and areas with deformation (folds) in the basal region.     

Efficacy of entomopathogenic nematodes in insect cadaver formulation against engorged females of Rhipicephalus microplus (Acari: Ixodidae) in semi-field conditions

The present study evaluated, in the laboratory, the virulence of Heterorhabditis bacteriophora LPP30 against engorged females of Rhipicephalus microplus; in addition, we evaluated, in semi-field tests, the effects of four isolates of Heterorhabditis spp. (i.e., H. bacteriophora HP88, Heterorhabditis baujardi LPP7, Heterorhabditis indica LPP1 and H. bacteriophora LPP30) in insect cadaver formulation against the non-parasitic phase of R. microplus. In the first experiment (in vitro), engorged females were exposed, in Petri dish, to H. bacteriophora LPP30 at 75, 150, 300, 600, 1200 and 2400 nematodes/tick (10 ticks per concentration tested). In the second experiment (semi-field), five engorged females and four cadavers of Galleria mellonella infected with nematodes were placed in pots with soil and grown Brachiaria decumbens; the pots were hold outdoor, exposed to natural environment conditions during the tests. In the third experiment (semi-field), 65 days after the cadavers had been placed in the pots for the second experiment, new engorged females (five per pot) were placed in the pots of the groups treated with H. bacteriophora HP88 or H. baujardi LPP7, to assess their persistence in the soil. In the first test, the percent control was higher than 95% in all treatment groups. In the second experiment, at day 22, the mean mortality rate was 78% in the groups treated with H. bacteriophora LPP30 or H. indica LPP1, and reached 100% and 98% when treated with H. bacteriophora HP88 and H. baujardi LPP7, respectively. In this experiment, the egg-laying inhibition index was higher than 90% in the groups treated with H. bacteriophora HP88 (97.2%) or H. baujardi LPP7 (91.9%). In the third experiment with H. bacteriophora HP88 and H. baujardi LPP7, the egg-laying inhibition index was 59.1% and 43.1%, respectively. We concluded that the isolate LPP30 was highly virulent under laboratory conditions, whereas in semi-field tests, HP88 and LPP7 were the most effective isolates. Moreover, HP88 and LPP7 remained active against engorged females of R. microplus in the soil for 65 days after application of EPN-infected cadavers of G. mellonella.     

Evaluation of cytotoxic effects of amitraz and fipronil on digestive,reproductive and neural processes of engorged Rhipicephalus microplus (Acari: Ixodidae) female

Fipronil and amitraz are potentially toxic compounds used for controlling ticks infesting pet and livestock. The use of fipronil on large animals was limited because of its high costs while amitraz is still persisting in the market since its introduction over four decades ago. Though resistance in ticks against these pesticides has been reported worldwide since 2000, the toxicity of these chemicals at cellular level in ticks is still poorly understood. The present study aimed to examine the gross and cellular impact of fipronil and amitraz on the gut, ovaries and synganglion of engorged Rhipicephalus microplus females. Fipronil and amitraz treated tick groups showed formation of a large number of vacuoles of different size throughout the cytoplasm of generative cells whereas sessile, residual and detached digestive cells were very low in numbers. The treatment of ticks resulted in the formation of vacuolations at periphery of all oocytes. Ultra-thin sections of the synganglion revealed severe rupture of neural lamella and perineurium with apoptosis of neural cells after fipronil treatment whereas in the amitraz treated ticks, severe destruction of neuropile region and extensive vacuolation of type I and II cells of cortical region as compared to the unexposed ticks were noted.     

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.     

First records of Ixodes brunneus (Acari: Ixodidae) in Delaware

Eight adult specimens of Ixodes brunneus (Acari: Ixodidae) were collected in Delaware via a combination of methods. The first was an engorged female recovered from a deceased Northern Mockingbird (Mimus polyglottos), a window strike victim, in Kent County. The other seven were four males and three females, all apparently unfed, collected by flagging and dragging vegetation in Kent, New Castle, and Sussex Counties during routine tick surveillance. These collections represent a new host record as well as new state and county records, and indicate a statewide presence of this species.     

Survival of off-host unfed Rhipicephalus (Boophilus) annulatus (Acari: Ixodidae) larvae in study arenas in relation to climatic factors and habitats in South Texas,USA

The cattle fever tick, Rhipicephalus annulatus (Say), is an economically destructive arthropod because of its ability to vector bovine babesiosis. Cattle fever ticks can spend more than 90% of their life cycle as questing larvae, but the effect of climatic factors on their off-host behavior and survival is unclear. The goal of this study was to measure the effects of specific ecological factors on off-host larvae in nature. The study was conducted in a south Texas pasture over a 20-mo period, during which time larval populations were surveyed and ambient weather variables - relative humidity and temperatures – were recorded. Oviposition success and larval survival varied between cattle fever tick cohorts and was affected by relative humidity and canopied (with tree cover) versus exposed habitat. The results show that relative humidity and the interaction of relative humidity and inhabiting canopied habitats play a key role in oviposition success. Additionally, canopied habitats have a positive influence on off-host larval survival in the spring and summer.     

Interaction between anti-tick vaccine and a macrocyclic lactone improves acaricidal efficacy against Rhipicephalus (Boophilus) microplus (Canestrini) (Acari: Ixodidae) in experimentally infested cattle

The southern cattle fever tick (SCFT) Rhipicephalus (Boophilus) microplus, is considered the most important ectoparasite of livestock in the world because of high financial losses associated with direct feeding and transmission of the hemoparasites Babesia bovis, B. bigemina, and Anaplasma marginale. Unfortunately, SCFT in many parts of the world have evolved resistance to all market-available pesticides thus driving development of new control technologies. Vaccination against ticks using the tick gut protein Bm86 has been shown to be effective against acaricide-resistant ticks. This technique has been successfully implemented in Puerto Rico for the control of acaricide-resistant R. microplus on dairy and beef cattle. Observations from Puerto Rico indicate a potentially positive interaction between anti-tick vaccination when used in conjunction with systemic acaricide treatment. In this project, controlled animal studies were completed directly comparing efficacy of anti-tick vaccination with and without systemic acaricide. Results show that the Bm86 anti-tick vaccine in combination with the macrocyclic lactone, Moxidectin, expressed a synergistic interaction, providing greater and longer efficacy than either treatment alone.     

Rhipicephalus capensis (Acari: Ixodidae), A geographically restricted South African tick,returning with a human traveler to the United States

Accelerated frequency of recreational travel, globalization of business, and legal and illegal plant and animal trades have contributed to enduring introductions of exotic ticks into the United States. We herein report a new incursion of a female Rhipicephalus capensis on a human traveler returning to Connecticut from South Africa. Natural distribution of R. capensis is restricted to the Western Cape Province and southwestern portion of the Northern Cape Province of South Africa, an area called the Fynbos Biome, and adults of this species primarily parasitize large, wild ungulate hosts. Only one previous international introduction of this tick is documented on imported South African plant material into the United States in 1985. The specimen described here was identified initially by morphological means and subsequently, a partial DNA sequence for the mitochondrial ribosomal RNA gene was generated in a PCR assay, which showed 94.86% identity to an R. capensis sequence in GenBank. We also provide information on several other previously unreported or under-reported incursions by South African ticks into the United States in association with imported Fynbos floricultural materials and speckled Cape tortoises, Chersobius signatus. Documentation of these additional exotic tick species incursions highlights ongoing challenges of the international movement of humans, animals, and other goods carrying ticks of human and veterinary importance.     

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.     

Species richness and adaptation of the invasive cattle tick,Rhipicephalus microplus (Acari: Ixodidae) on camps grazed by sheep in the Eastern Cape Province,South Africa

The seasonal dynamics and distribution of ticks play a pivotal role in the epidemiology of tick-borne diseases. The objective of this study was to establish the seasonal abundance and species richness of questing ixodid ticks on Amathole Montane Grassland camps grazed by sheep. Ticks questing for hosts were collected monthly for a period of three years by dragging flannel strips attached to a wooden spar over the vegetation. At each occasion, six replicate drag-samples were made in camps grazed by sheep. Of the questing ticks (n = 14 891) collected from the vegetation, the most abundant larvae were those of Rhipicephalus microplus (95.04%) followed by Rhipicephalus appendiculatus (2.32%), Rhipicephalus evertsi (1.56%), Rhipicephalus decoloratus (1.03%), Rhipicephalus simus (0.03) and Amblyomma hebraeum (0.02%). Comparing the two Rhipicephalus (Boophilus) spp., R. microplus (98.9%) outcompete the indigenous tick, R. decoloratus (1.1%). The R. microplus larvae were significantly higher (P < 0.05) in 2015 (2.11±0.108), 2016 (2.02 ± 0.076) and 2017 (1.94±0.075) during spring than any other season. There were no significant differences (P ? 0.05) from R. appendiculatus questing ticks collected in autumn (0.27±0.007; 0.30±0.052) and spring (0.33±0.007; 0.20±0.052) for 2015 and 2016, respectively. The study showed that the cattle tick, R. microplus adapted very well on host species, in this case sheep, and encroached to areas that were too cold for its adaptation.     

The life cycle of Hyalomma scupense (Acari: Ixodidae) under laboratory conditions

The life cycle of Hyalomma scupense on rabbit hosts was investigated under laboratory conditions. Hy. scupense exhibited one- and two-host life cycles of 163.2 and 161.4 days, respectively. The incubation of eggs required an average period of 52 days, which was the longest period among the four developmental stages. The average time for pre-feeding of larvae was 3.5 days. It took 20 days for larvae to become engorged nymphs and 52.3 days to become engorged females. The duration of the pre-feeding, feeding, pre-oviposition, and oviposition stages of female adults was 2.3, 13.5, 27.5, and 27.9 days, respectively. The average weight of engorged females was 390.0 mg (ranging from 129.3 mg to 828.6 mg), which was 28.95 times the weight of unfed females. There was a positive relationship between the weight and the number of eggs laid by engorged females (r = 0.927). The reproductive efficiency index (REI) was 8.63.     

Associations between wild birds and hard ticks (Acari: Ixodidae) in Colombia

Ticks of the family Ixodidae are vectors of important pathogens in human and animal health. Birds are involved in long-distance transport and dispersion of hard ticks. Tick infestations on wild birds mostly involve species within the genera Amblyomma, Ixodes, and Haemaphysalis. In Colombia, tick research is scarce and there are no studies to date about the associations between wild birds and ticks. We aimed to contribute to the knowledge of the associations between wild birds and hard ticks based on the collection of 2314 wild birds belonging to 29 families in Caldas – Colombia between 2015 and 2019. In total, we collected 133 hard ticks that were found parasitizing 78 birds representing 45 species and 14 wild bird families. We report at least seven tick species on birds confirmed by morphological and molecular methods: Amblyomma longirostre, Amblyomma varium, Amblyomma dissimile, Amblyomma ovale, Amblyomma nodosum, Amblyomma calcaratum and Haemaphysalis leporispalustris. In addition, we recorded three Ixodes species, which yielded DNA sequences that did not have high identity (≤ 95 %) to any species in GenBank. Ticks were found infesting resident and migratory boreal birds. This is the first study addressing the associations between wild birds and hard ticks in Colombia. We describe new associations between birds and ticks in the Americas.     

Synonymy of Ixodes aragaoi with Ixodes fuscipes,and reinstatement of Ixodes spinosus (Acari: Ixodidae)

Ixodes fuscipes Koch, 1844, described from Brazil, has been considered a valid species with records from Brazil and Peru. Ixodes spinosus Neumann, 1899, also described from Brazil, has been considered a synonym of I. fuscipes. In 2014, Ixodes aragaoi Fonseca, 1935 was redescribed as a valid species for Brazil and Uruguay. Until the present study, one major difference between the females of I. fuscipes and I. aragaoi was the horn-like auriculae in the former versus the ridge-like auriculae in the later, but this morphological difference was not supported after examination of the holotype of I. fuscipes and a syntype female of I. spinosus. Surprisingly, we found the type of I. fuscipes to have ridge-like auriculae, in contrast to the horn-like auriculae of I. spinosus. Comparisons of the I. fuscipes holotype with the syntypes of I. aragaoi revealed that they correspond to the same species. Therefore, we redescribe I. fuscipes, relegate I. aragaoi to a junior synonym of I. fuscipes, and reinstate I. spinosus as a valid species. After examining all lots of I. fuscipes in four tick collections, no specimen was recognized as I. fuscipes when compared with the type specimen of this taxon. On the other hand, specimens previously identified as I. aragaoi are now confirmed as I. fuscipes, with bona fide records for Brazil and Uruguay. Some of the specimens previously reported as I. fuscipes are now confirmed as I. spinosus, with records in two Brazilian biomes, Amazon and Atlantic rainforest. We present lists of the ticks examined in this study, with their current taxonomic status.     

Pathogens in Ixodes persulcatus and Ixodes ricinus ticks (Acari,Ixodidae) in Karelia (Russia)

Ixodid ticks (Acarina, Ixodidae) are vectors of dangerous human infections. The main tick species that determine the epidemiological situation for tick-borne diseases in northern Europe are Ixodes ricinus and Ixodes persulcatus. In recent years, significant changes in the number and distribution of these species have been observed, accompanied by an expansion of the sympatric range. This work summarizes the data of long-term studies carried out in Karelia since 2007 on the infection of I. persulcatus and I. ricinus ticks with various pathogens, including new viruses with unclear pathogenic potential. As a result, tick-borne encephalitis virus (TBEV, Siberian genotype), Alongshan virus, several representatives of the family Phenuiviridae, Borrelia afzelii, Borrelia garinii, Ehrlichia muris, Candidatus Rickettsia tarasevichiae and Candidatus Lariskella arthropodarum were identified. Data were obtained on the geographical and temporal variability of tick infection rates with these main pathogens. The average infection rates of I. persulcatus with TBEV and Borrelia burgdorferi sensu lato were 4.4% and 23.4% and those of I. ricinus were 1.1% and 11.9%, respectively. We did not find a correlation between the infection rate of ticks with TBEV, B. burgdorferi s.l. and Ehrlichia muris/chaffeensis with the sex of the vector. In general, the peculiarities of the epidemiological situation in Karelia are determined by the wide distribution and high abundance of I. persulcatus ticks and by their relatively high infection rate with TBEV and B. burgdorferi s.l. in most of the territory, including the periphery of the range.     

Phylogenetic relationships of the Amblyomma cajennense complex (Acari: Ixodidae) at mitogenomic resolution

The genus Amblyomma is the third most diverse in the number of species within the Ixodidae, with practically half of its species distributed in the Americas, though there are also species occurring in Africa, Asia, and Australia. Within the genus, there are several species complexes with veterinary and public health importance. The Amblyomma cajennense complex, in the Americas, is represented by six species with a wide distribution, from Texas to northern Argentina. We combined two sequencing techniques to generate complete mitogenomes of species belonging to the Amblyomma cajennense complex: genome skimming and long-range PCRs sequencing methods. Thus, we generated seven new mitochondrial genomes for all species of the Amblyomma cajennense complex, except for Amblyomma interandinum. Genetic distances between the mitogenomes corroborate the clear differentiation between the five species of the Amblyomma cajennense complex. The phylogenetic relationships of these species had previously been evaluated by combining partial nuclear and mitochondrial genes and here these relationships are corroborated with a more robust framework of data, which demonstrates that the conjunction of mitochondrial and nuclear partial genes can resolve close relationships when entire genes or genomes are unavailable. The gene order, structure, composition, and length are stable across these mitogenomes, and they share the general characteristics of Metastriata. Future studies should increase the number of available mitogenomes for this genus, especially for those species from the Indo-Pacific region and Africa, by means of a better understanding of their relationships and evolutionary process.