Acquired tick resistance (ATR) is well characterized in tick-exposed animals, compromising tick fitness through antibody-mediated activation of basophils. Yet, anti-tick vaccines inducing ATR have had limited success. Here, we describe a neuroimmune event preceding ATR that leads to rapid host-mediated tick removal. Tick-sensitized guinea pigs mechanically remove ticks within 3-6 hours via an acquired neuroimmune-induced itch response that correlates with increased dermal expression of itch-associated genes like OSM and skin infiltration by T cells and macrophages, independently of IgG and IgE antibodies. When we expose humans to ticks, a similar immune response is observed. Blocking T cells before tick sensitization prevents immune cell infiltration to bite sites and abrogates scratching and tick removal. This neuroimmune response is independent of Trpv1 as tick-sensitized guinea pigs treated with resiniferatoxin remove ticks effectively. Itch-induced tick removal or IITR offers a novel approach to tick-borne disease prevention through early tick detection and removal.

The insect molting process is critical to growth and development and is regulated in part by the neuropeptides corazonin, eclosion hormone, and α and β bursicon. We found messages in a synganglion transcriptome from adult, female American dog ticks, Dermacentor variabilis (that do not molt), with a high similarity to the larval insect neuropeptides that control molting. The phylogenetic analysis of the tick putative neuropeptides compared to other arthropods is discussed in detail. The relative gene expression of these peptides was determined by quantitative PCR during the following adult developmental stages: (i) virgin, unfed 0–24 h after entering the adult stage (non-host-seeking), (ii) host-seeking, unfed, and not mated (3 d after emergence), (iii) part-fed (unmated, attached to host; 1st and 3rd day after emergence), (iv) mated (females are part-fed; allowed to mate for ≤1 day, 7th day after emergence), (v) mated repletes (completion of blood feeding but still attached to host), and (vi) post-drop-off (from host) with egg laying starting within 1 d of detachment. Eclosion hormone transcript levels peaked at mating and at drop-off. Bursicon α levels were highest just after molting into adults, with a second smaller peak in replete females. Bursicon β levels were highest (32-fold) post-drop-off. Corazonin message levels peaked in part-feds and were much higher (40-fold) in repletes compared to 0–24 h after emergence. RNAi suppression of the corazonin message by injection in newly molted ticks reduced oviposition and the number of vitellogenic eggs in the ovaries at drop-off but had no apparent effect on host-seeking, partial feeding, mating, feeding to repletion, and drop-off. The possible roles of these transcripts in adult, female tick development are discussed.

Tick-borne diseases pose significant risks to both animals and humans, with emerging pathogens like Ehrlichia muris eauclairensis (EME) underscoring the need for a deeper understanding of pathogen-vector interactions and tick fitness. This study investigates the impact of EME on Ixodes scapularis nymphs, revealing significant behavioral changes in EME-positive ticks. These ticks exhibited increased movement speed, faster bite site-seeking for attachment, and prolonged feeding durations compared to control ticks. Proteomic analyses of the tick synganglion during resting and feeding phases identified 196 differentially expressed proteins in EME-positive ticks, including multiple proteins associated with nicotinic acetylcholine signaling pathways. Our findings indicated altered neuropeptide expression related to stimulus response and activity, suggesting changes in neurophysiology. This research provides the first evidence of behavioral manipulation by an Ehrlichia species, indicating that the tick nervous system is a site of bacterial influence and a potential target for interventions. These findings offer new insights into pathogen-vector dynamics that could lead to the development of transmission-blocking therapies, significantly impacting tick fitness and disease transmission.IMPORTANCETick-borne diseases (TBDs) are increasingly affecting humans, pets, and livestock, with cases rising in recent years. Ticks can carry multiple harmful germs, and human activities and environment are contributing to new TBDs. This study shows that the bacteria Ehrlichia muris eauclairensis (EME) can change the behavior of nymphal black-legged ticks, which spread various diseases. Infected ticks moved faster, attached to hosts more quickly, and fed longer than uninfected ticks. These changes were linked to specific proteins in the tick's nervous system, suggesting that EME manipulates tick behavior. This is the first evidence that an Ehrlichia species can influence tick behavior, potentially increasing disease transmission. Understanding these interactions can help develop strategies to prevent TBDs by targeting the bacteria's influence on ticks, ultimately reducing disease spread and improving public health.

A survey was conducted of microbes and arthropods associated with Varroa destructor (Acari: Varroidae) mites (hereafter, Varroa) collected from honey bee colonies in Maryland, USA. Live Varroa were challenged with samples of both microbes and arthropods and assessed for their potential as biocontrol agents against the mites. To our knowledge, this is the first survey of natural enemies of Varroa conducted in North American honey bee colonies. The survey revealed 21 fungi, 25 bacteria, one nematode, and 8 arthropod species associated with Varroa in honey bee colonies, and from challenge assays using 11 of these, including 4 species of fungi, 2 species of bacteria, a parasitic nematode, 2 insects (Hemiptera and Hymenoptera), and 2 arachnids (Pseudoscorpiones) were capable of killing Varroa. This assemblage of Varroa natural enemies was similar to taxa identified from surveys conducted elsewhere globally, suggesting there is a core assemblage of organisms capable of killing Varroa that are best suited to tolerate the sometimes-hostile abiotic and biotic conditions of the hive environment. Generally, entomopathogenic fungi and bacteria assessed in our study showed the highest efficacy against Varroa, and for some, efficacy was comparable to that determined by other researchers, suggesting entomopathogenic fungi and bacteria appear to be the most promising candidates for development into commercially viable varroicides. A novel parasitic nematode identified from our survey killed greater than 90% of infected hosts. In addition, the behaviours of some arthropods toward Varroa observed in our study were indicative of predation on the mites.

Amblyomma maculatum Koch (Acari: Ixodidae), Ixodes scapularis Say, and Dermacentor variabilis Say are three hard-bodied ticks responsible for vectoring pathogens that cause most human tick-borne diseases in the United States of America (USA). Tick surveillance is critical to elucidate high-risk areas for targeted vector control and public health interventions. Despite tick-borne diseases having a higher annual incidence compared to mosquito-borne diseases in the USA, tick traps used for surveillance are grossly under-developed compared to the suite of sophisticated mosquito traps available on the commercial market. This study sought to identify potential prospective tick trap improvements by evaluating the relative attraction of three medically important tick species to odor attractants in a laboratory setting: carbon dioxide, ammonia, deer musk, and rodent musk. This study found that carbon dioxide gas and rodent musk had limited attraction to all three tick species and across life stages. Conversely, this study did find that deer musk and ammonia demonstrated favorable attraction for I. scapularis and D. variabilis across multiple life stages, suggesting standard tick trap techniques might be improved using low-cost and practical alternate attractants. Future field-based studies of these odor attractants, both singularly and in combination with the standard dry ice, are warranted to evaluate potential increased efficacy for broad tick species vector surveillance methods.

Infections caused by vector-borne pathogens impose a significant burden of morbidity and mortality in a global scale. In their quest for blood, hematophagous arthropods penetrate the host skin and may transmit pathogens by the bite. These pathogens are deposited along with saliva and a complex mixture of vector derived factors. Hematophagous arthopod vectors have evolved a complex array of adaptations to modulate the host immune response at the bite site with the primary goal to improve blood feeding, which have been exploited throughout evolution by these pathogens to enhance infection establishment in the host. While this paradigm has been firmly established in mouse models, comparable data from human studies are scarce. Here we review how the host skin immune response to vector bites in animal models is hijacked by microbes to promote their pathogenesis. We mainly explored four distinct vector-pathogen pairs of global health importance: sand flies and Leishmania parasites, Ixodes scapularis ticks and Borrelia burgdorferi, Aedes aegypti mosquitoes and arboviruses, and Anopheles gambiae mosquitos and Plasmodium parasites. Finally, we outline how critical it is for the field of vector biology to shift from rodent models to clinical studies focused on the interface of vector-pathogen-host immune system to push further the frontiers of knowledge of the field.

Ticks are significant ectoparasites that transmit a variety of pathogens, leading to serious human and animal diseases, including Lyme disease, Rocky Mountain spotted fever, anaplasmosis and many others. The emergence of acaricide resistance in hard ticks presents a formidable challenge for public health and livestock management, exacerbated by the increasing incidence of tick-borne diseases and associated economic losses, estimated at $20 billion annually in the livestock sector alone. This review examines the mechanisms underlying acaricide resistance, focusing on genetic mutations, metabolic detoxification processes, and behavioral adaptations in tick populations. We detail the role of commercial acaricides in tick control while emphasizing the adverse effects of their overuse, which contributes to the development of resistant strains. Innovative control strategies are explored, including using pesticide synergists that enhance the efficacy of existing acaricides by targeting the tick's phosphagen system. Additionally, this review highlights the importance of understanding the synergistic interactions between various control methods, including non-chemical approaches such as personal protection measures and landscape management. The review concludes by underscoring the urgent need for novel acaricides with new modes of action and implementing regular monitoring practices to combat acaricide resistance effectively. Addressing these challenges is vital for the sustainable management of tick populations and protecting public health and livestock productivity.

Subolesin is a conserved molecule in both hard and soft ticks and is considered as an effective candidate molecule for the development of anti-tick vaccine. Previous studies have reported the role of subolesin in blood feeding, reproduction, development, and gene expression in hard ticks. However, studies addressing the role of subolesin in soft ticks are limited. In this study, we report that subolesin is not only important in soft tick Ornithodoros turicata americanus blood feeding but also in the regulation of innate immune gene expression in these ticks. We identified and characterized several putative innate immune genes including Toll, Lysozyme precursor (Lp), fibrinogen-domain containing protein (FDP), cystatin and ML-domain containing protein (MLD) in O. turicata americanus ticks. Quantitative real-time polymerase chain reaction analysis revealed the expression of these genes in both O. turicata americanus salivary glands and midgut and in all developmental stages of these soft ticks. Significantly increased expression of fdp was noted in salivary glands and midgut upon O. turicata americanus blood feeding. Furthermore, RNAi-mediated knockdown of O. turicata americanus subolesin expression affected blood feeding and innate immune gene expression in these ticks. Significant downregulation of toll, lp, fdp, cystatin, and mld transcripts was evident in sub-dsRNA-treated ticks when compared to the levels noted in mock-dsRNA-treated control. Collectively, our study not only reports identification and characterization of various innate immune genes in O. turicata americanus ticks but also provides evidence on the role of subolesin in blood feeding and innate immune gene expression in these medically important ticks.

Varroa destructor is a cosmopolitan pest and leading cause of colony loss of the European honey bee. Historically described as a competent vector of honey bee viruses, this arthropod vector is the cause of a global pandemic of Deformed wing virus, now endemic in honeybee populations in all Varroa -infested regions. Our work shows that viral spread is driven by Varroa actively switching from one adult bee to another as they feed. Assays using fluorescent microspheres were used to indicate the movement of fluids in both directions between host and vector when Varroa feed. Therefore, Varroa could be in either an infectious or naïve state dependent upon the disease status of their host. We tested this and confirmed that the relative risk of a Varroa feeding depended on their previous host’s infectiousness. Varroa exhibit remarkable heterogeneity in their host-switching behavior, with some Varroa infrequently switching while others switch at least daily. As a result, relatively few of the most active Varroa parasitize the majority of bees. This multiple-feeding behavior has analogs in vectorial capacity models of other systems, where promiscuous feeding by individual vectors is a leading driver of vectorial capacity. We propose that the honeybee- Varroa relationship offers a unique opportunity to apply principles of vectorial capacity to a social organism, as virus transmission is both vectored and occurs through multiple host-to-host routes common to a crowded society.

Artificial membrane feeding (AMF) is a powerful and versatile technique with a wide range of applications in the study of disease vectors species. Since its first description, AMF has been under constant optimization and standardization for different tick species and life stages. In the USA, Ixodes scapularis is the main vector of tick-borne zoonoses including the pathogens causing Lyme disease in humans and animals. Seeking to improve the overall fitness of I. scapularis adult females fed artificially, here, we have optimized the AMF technique, considerably enhancing attachment rate, engorgement success, egg laying, and egg hatching compared to those described in previous studies. Parameters such as the membrane thickness and the light/dark cycle to which the ticks were exposed were refined to more closely reflect the tick’s natural behavior and life cycle. Additionally, ticks were fed on blood only, blood + ATP or blood + ATP + gentamicin. The artificial feeding of ticks on blood only was successful and generated a progeny capable of feeding naturally on a host, i.e., mice. Adding ATP as a feeding stimulant did not improve tick attachment or engorgement. Notably, the administration of gentamicin, an antibiotic commonly used in tick AMF to prevent microbial contamination, negatively impacted Rickettsia buchneri endosymbiont levels in the progeny of artificially fed ticks. In addition, gentamicin-fed ticks showed a reduction in oviposition success compared to ticks artificially fed on blood only, discouraging the use of antibiotics in AMF. Overall, our data suggest that the AMF of adult females on blood only, in association with the natural feeding of their progeny on mice, might be used as an integrated approach in tick rearing, eliminating the use of protected species under the Animal Welfare Act (AWA). Of note, although optimized for I. scapularis adult ticks, I. scapularis nymphs, other tick species, and sand flies could also be fed using the membrane described in this study, indicating that it might be a suitable alternative for the artificial feeding of a variety of hematophagous species.

EDITORIAL article Front. Cell. Infect. Microbiol., 17 March 2022Sec.Parasite and Host https://doi.org/10.3389/fcimb.2022.885240

The rapid reproductive capacity of Varroa destructor is among the most significant adaptations underpinning its success as a parasite. To exploit their honey bee host, the parasite must rapidly produce offspring that fully develop into adults and mate in an inflexible 9-day window. Inability to meet this deadline brings the foundress mite’s fitness to zero establishing heavy evolutionary pressure to accelerate reproduction & subsequent development of offspring. Our work fills in gaps in our understanding of a key pathway in this process. Varroa have a poorly-understood ability to pass heretofore unidentified host proteins through their body with minimal digestion/degradation. Via Native-PAGE, we were able to confirm that nine proteins shared with honey bee fat body tissue accumulate intact in the eggs of the foundress mite. As such, we hypothesized that the proteins were several egg yolk precursors synthesized and stored in the fat body. Using antibodies raised against honey bee vitellogenin (Vg) we positively identified this egg yolk precursor via SDS-PAGE and subsequent Western Blot. We then analyzed samples of honey bee fat body tissue, gravid Varroa , so called “phoretic” Varroa , and Varroa eggs via HPLC MS/MS to identify the remaining host proteins and determine their relative abundance. We detected egg yolk precursors in the large lipid transfer protein superfamily, in addition to hexamerin storage proteins, and miscellaneous motor/transfer proteins integral to embryonic development (transferrin, myosin heavy chain, and heat shock protein 60). Varroa lack the capacity to produce some of these proteins and instead employ kleptoparasitism on a molecular level to provision their developing ova, a pathway not described in any other host/parasite relationship, hereafter referred to as kleptocytosis. These different families of proteins are normally produced by the female and conveyed to the vitellogenic egg cell through protein specific receptor-mediated pathways. Such pathways would exclude foreign proteins. We hypothesize that the need to rely on a receptor-mediated pathway is circumvented via the specialized nutritive reproductive tissue, the lyrate organ. Through microCT imaging we detail the connection between the developing ovum and this dual-lobed organ. Better understanding of this pathway presents a novel target for Varroa management as the treatment need only accomplish slowing acquisition or deposition of host proteins thereby disrupting the mite’s ability to meet the temporal demand of its host.

This expert opinion provides a brief review of Anaplasma phagocytophilum growth in its tick host as well as evidence on the impact of climate change on tick and tick-borne disease persistence and spread.

Background Ixodes scapularis is the predominant tick vector of Borrelia burgdorferi , the agent of Lyme disease, in the USA. Molecular interactions between the tick and B. burgdorferi orchestrate the migration of spirochetes from the midgut to the salivary glands—critical steps that precede transmission to the vertebrate host. Over the last decade, research efforts have invoked a potential role for the tick microbiome in modulating tick-pathogen interactions. Results Using multiple strategies to perturb the microbiome composition of B. burgdorferi -infected nymphal ticks, we observe that changes in the microbiome composition do not significantly influence B. burgdorferi migration from the midgut, invasion of salivary glands, or transmission to the murine host. We also show that within 24 and 48 h of the onset of tick feeding, B. burgdorferi spirochetes are within the peritrophic matrix and epithelial cells of the midgut in preparation for exit from the midgut. Conclusions This study highlights two aspects of tick-spirochete interactions: (1) environmental bacteria associated with the tick do not influence spirochete transmission to the mammalian host and (2) the spirochete may utilize an intracellular exit route during migration from the midgut to the salivary glands, a strategy that may allow the spirochete to distance itself from microbiota in the midgut lumen effectively. This may explain in part, the inability of environment-acquired midgut microbiota to significantly influence spirochete transmission. Unraveling a molecular understanding of this exit strategy will be critical to gain new insights into the biology of the spirochete and the tick.

Bacterial endosymbionts are abundantly found in both hard and soft ticks. Occidentia massiliensis, a rickettsial endosymbiont, was first identified in the soft tick Ornithodoros sonrai collected from Senegal and later was identified in a hard tick Africaniella transversale. In this study, we noted the presence of Occidentia species, designated as Occidentia-like species, in a soft tick O. turicata americanus. Sequencing and phylogenetic analyses of the two genetic markers, 16S rRNA and groEL confirmed the presence of Occidentia-like species in O. turicata americanus ticks. The Occidentia-like species was noted to be present in all developmental stages of O. turicata americanus and in different tick tissues including ovaries, synganglion, guts and salivary gland. The levels of Occidentia-like species 16S rRNA transcripts were noted to be significantly higher in ovaries than in a gut tissue. In addition, Occidentia-like species groEL expression was noted to be significantly higher in tick synganglion than in ovaries and gut tissues. Furthermore, levels of Occidentia-like species 16S rRNA transcripts increased significantly upon O. turicata americanus blood feeding. Taken together, our study not only shows that Occidentia-like species is present in O. turicata americanus but also suggests that this bacterium may play a role in tick-bacteria interactions.

Background: Blood-feeding arthropods support a diverse array of symbiotic microbes, some of which facilitate host growth and development whereas others are detrimental to vector-borne pathogens. We found a common core constituency among the microbiota of 16 different arthropod blood-sucking disease vectors, including Bacillaceae, Rickettsiaceae, Anaplasmataceae, Sphingomonadaceae, Enterobacteriaceae, Pseudomonadaceae, Moraxellaceae and Staphylococcaceae. By comparing 21 genomes of common bacterial symbionts in blood-feeding vectors versus non-blooding insects, we found that certain enteric bacteria benefit their hosts by upregulating numerous genes coding for essential nutrients. Bacteria of blood-sucking vectors expressed significantly more genes (p < 0.001) coding for these essential nutrients than those of non-blooding insects. Moreover, compared to endosymbionts, the genomes of enteric bacteria also contained significantly more genes (p < 0.001) that code for the synthesis of essential amino acids and proteins that detoxify reactive oxygen species. In contrast, microbes in non-blood-feeding insects expressed few gene families coding for these nutrient categories. We also discuss specific midgut bacteria essential for the normal development of pathogens (e.g., Leishmania) versus others that were detrimental (e.g., bacterial toxins in mosquitoes lethal to Plasmodium spp.).

Extracellular vesicles are thought to facilitate pathogen transmission from arthropods to humans and other animals. Here, we reveal that pathogen spreading from arthropods to the mammalian host is multifaceted. Extracellular vesicles from Ixodes scapularis enable tick feeding and promote infection of the mildly virulent rickettsial agent Anaplasma phagocytophilum through the SNARE proteins Vamp33 and Synaptobrevin 2 and dendritic epidermal T cells. However, extracellular vesicles from the tick Dermacentor andersoni mitigate microbial spreading caused by the lethal pathogen Francisella tularensis . Collectively, we establish that tick extracellular vesicles foster distinct outcomes of bacterial infection and assist in vector feeding by acting on skin immunity. Thus, the biology of arthropods should be taken into consideration when developing strategies to control vector-borne diseases.

Histochemical staining of histological sections of Varroa destructor () mites reveal the internal body plan and are used to contrast the internal organs associated with feeding and reproduction of starved versus recently fed female mites. The gnathosoma is comprised of a powerful sucking pharynx, which employs 11 alternating dilator and constrictor muscles, the chelicerae, the salivary ducts, and the salivarium. Coronally, the esophagus is visible through the synganglion between its supraesophageal and subesophageal regions and connects posteriorly to the midgut. The midgut is devoid of food particles in starved mites, but in fed mites, the midgut epithelial cells are filled with innumerable globular spheroids replete with lipoproteins, including polyunsaturated lipids, whereas the lumen is filled with saturated lipids or other unidentified nutrients. In the opisthosomal body region of the fed female, the bilobed lyrate organ lies adjacent to the midgut on one side and the ovary on the opposite side, with very close cell to cell linkages that appear to form a syncytium. The fed female ovary contains an enormously enlarged ovum, and numerous elongated nurse cells extending from the lyrate organ. Dyes staining selectively for lipoproteins suggests rapid incorporation of neutral and polyunsaturated lipids and lipoproteins. Also evident near the ovary in fed females is the spermatheca filled with elongated, fully capacitated spermatozoa. The histological and histochemical findings reported in this study provide a fresh insight into the body structure, nutrition, and reproductive activity of the female of this harmful honey bee parasite and disease vector.

Ixodes scapularis ticks transmit multiple pathogens, including Borrelia burgdorferi sensu stricto, and encode many proteins harboring epidermal growth factor (EGF)-like domains. We show that I. scapularis produces multiple orthologs for Bm86, a widely studied tick gut protein considered as a target of an anti-tick vaccine, herein termed as Is86. We show that Is86 antigens feature at least three identifiable regions harboring EGF-like domains (termed as EGF-1, EGF-2, and EGF-3) and are differentially upregulated during B. burgdorferi infection. Although the RNA interference-mediated knockdown of Is86 genes did not show any influences on tick engorgement or B. burgdorferi sensu stricto persistence, the immunization of murine hosts with specific recombinant EGF antigens marginally reduced spirochete loads in the skin, in addition to affecting tick blood meal engorgement and molting. However, given the borderline impact of EGF immunization on tick engorgement and pathogen survival in the vector, it is unlikely that these antigens, at least in their current forms, could be developed as potential vaccines. Further investigations of the biological significance of Is86 (and other tick antigens) would enrich our knowledge of the intricate biology of ticks, including their interactions with resident pathogens, and contribute to the development of anti-tick measures to combat tick-borne illnesses.

Ticks exist across diverse environments and transmit numerous pathogens. Due to their long and unique life cycles, these arthropods likely evolved robust epigenetic mechanisms that provide sustainable responses and buffers against extreme environmental conditions. Herein, we highlight how the study of the epigenetic basis of tick biology and vectorial capacity will enrich our knowledge of tick-borne infections.