Attachment Structure of Wood Ticks; a Fine Structure Study
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Heckmann Pakistan Journal of Parasitology 65; June 2018 ATTACHMENT STRUCTURE OF WOOD TICKS; A FINE STRUCTURE STUDY Richard Heckmann Department of Biology - 1114 MLBM, Brigham Young University, Provo,Utah 84602 USA Abstract: The mouthparts of four species of ticks (Rhipicephalus, Amblyomma, Dermacenter and Haemaphysalis) were viewed with SEM and compared to one species of mite (Varrao). The ticks have the characteristic pedipalps, chelicera and hypostome to attach and feed on hosts. The appendage modification for host attachment was viewed including the pulvilli and Haller’s organ. Specific mouthparts were scanned with x-ray (XEDS) and the barbs of the hypostome were cut with a gallium ion beam (LIMS). For comparison, the mouthparts of a mite (Varrao) were included in the study. Those organisms studied belonged to the Acarina. Keywords: Ticks, Mites, Mouthparts, Hypostome, Acarina. INTRODUCTION The mouthparts of a tick are designed for puncture of a host skin and then feed on the host body fluids, especially blood. During the feeding process the eight leg Acarinid can transmit many diseases may acquired by human hosts. Birds, mammals and reptiles are infected with blood feeding ticks. (Macnair, 2016) reported that approximately 850 species have been described worldwide (Wikipedia). Tick, 2018 and Sonenshine, 1991 had suggested that ticks are important vector of disease in humans and animals. Mouthparts of ticks have three visible components for mouthparts. The two outside parts which are joined are highly mobile palps also called (pedipalps) in between these are paired chelicerae which protect the hypostome. While ticks are deeding the palps more laterally. Beak-like projections are present or the bared hyperstome, this structure plunges while feeding on host skin. Moreover, most hard ticks are capable of secreting a cement like substance which while feeding glues the tick. This substance once feeding is complete is dissolved. (Vredevoe, 2017; Sonenshine, 1991; Bishop et al., 2002 and Betz, 1996). Parts of the gnathostome represent other parts of the tick body. The objectives of this paper were to demonstrate using scanning electron microscopy (SEM) the mouthparts, or feeding mechanisms of four species of Ixodidae namely, Haemaphysalis, Rhipicephalus, Dermacenter, and Amblyomma and comment on the mouthparts of other hard- body ticks. The second goal was to show the structures critical for initial attachment and 7 Heckmann Pakistan Journal of Parasitology 65; June 2018 sensing of a tick, with subsequent feeding of the ectoparasite. These tick mouthparts will be compared to a mite (Varrao) which is closely related to the ticks. The Varrao is destructive to honeybees (Apis). Description of the four ectoparasites is as follows; (Sonenshine, 1991). Rhipicephalus, is a common tick throughout the world. One species is known as the dog tick (R. sanguineus). It is more common in warmer climates. It can complete its lifecycle indoors. There are numerous species of this genus, which are cosmopolitan in distribution. Amblyomma is the third largest tick in the number of species and distribution for the Ixodidae. There are many species, which are widely distributed and account for anaplasmosis and in Brazil Rocky Mountain spotted fever. Haemaphysalis a common hard-body tick (Ixodidae) found on many mammalian hosts both domestic and wild, worldwide. Also on birds and reptiles. Vector of many diseases including Lyme disease, tick paralysis, tularemia, Q fever, etc. Varrao is a mite that infects honeybees and feeds on the endolymph of the host. Like ticks, it has 8 appendages and biting mouthparts with prominent chelicerae. They are effective vectors of virus and bacteria diseases found in honey bees. Dermacenter: A hard body tick with 3 life stages (larvae, nymph and adult). Most species such as D. andersoni generally are found in North West United States and South West Canada along the Rocky Mountains. It is a vector for many diseases (Colorado Tick fever, Tularemia, Rocky Mountain spotted fever). The female can feed for up to 5-15 days, thus ticks must be immediately removed (Manyarubuga, 2012). Ticks are small acarinids, part of the order Parasitiformes. Along with mites, they constitute the subclass Acari. Ticks are ectoparasites living by feeding on the blood of mammals, birds, and sometimes reptiles and amphibians. Moreover, the ticks belong to a family Argasidae and Ixodidae. Ticks and mites have lost abdomen segmentation which their ancestors possessed leading to fusion of abdomen with the cephalothorax. They lack a head with eyes and brain. The tagmata typical of other Chelicerae have been replaced by two new body sections, the anterior capitulum (or gnathosoma), which is retractable and contains the mouthparts, and the posterior idiosoma which contains the legs, digestive tract, and reproductive organs. For piercing skin and blood sucking capitulum is used; it is only in the front of the head and contains neither the brain nor the eyes (Wikipedia, 2018; and Sonenshine, 1991). This study emphasized the mouthparts of Ticks. Major modifications are present on the appendages of the Tick including pulvilli and Haller’s organ, along with a questing activity aiding in the host attachment and detection. 8 Heckmann Pakistan Journal of Parasitology 65; June 2018 MATERIALS AND METHODS Specimens of both ticks (Haemaphysalis, Rhipicephalus, Dermacenter and Amblyomma) and mites (Varrao, honeybee mite) were obtained and fixed in 70% ethyl alcohol. SEM (Scanning Electron Microscopy) The ticks were fixed in 70 percent ethanol and processed according to Lee, 1992, which included critical point drying (CPD) is sample baskets and subsequently mounting on SEM sample mounts (stubs) with the help of double-sided carbon tape. Samples were coated with gold and palladium for 3 min using a Polaron #3500 sputter coater (Quorum (Q150 TES) www.quorumtech.com) establishing an approximate thickness of 20 nm. Samples were placed and observed in an FEI Helios 660 NanoLab DualBeam (FEI, Hillsboro, OR) scanning electron microscope, with digital images obtained in the Nanolab software system (FEI, Hillsboro, OR), and later transferred to a USB for future reference. Moreover, the images were taken at various magnifications. The samples were received under low vacuum conditions with 10 KV, spot size 2, 0.7 Torr using a GSE detector. RESULTS Figures 1 to 12 (Plate 1 and 2) represent the SEM results for the wood ticks mainly Rhipicephalus and Dermacenter while figures 13-16 (Plate 3) are SEM scans of the honeybee mite, Varrao destructor. The gallium cuts of the barbs of the hypostome (Fig. 6A and 6B) are represented by Fig. 10. The gallium cut results will be part of another article. Pedipalps, chelicerae, and hypostome are visible (Figs. 1 to 5, 8, 9). The hypostome with its barbs is very prominent (Figs. 1 and 2). The initial cutting appendage, chelicerae, shows prominently for Figs. 4, 5 and 6. The intact mouthparts show up for Fig. 8 for another tick species. The cut barb of a hypostome (Fig. 9) is displayed by Figure 10. Part of the host tissue surrounded by the tick mouthparts is given with Fig. 7. Figs. 11A, 11B and 12 display the unique pulvillus near Haller’s Organ. Note the claw-like structure on the terminal segment of the appendage (Figs. 11A, 11B and 12). Plate 3 (Figs. 13-16) are SEM micrographs of the Varrao mite. Figs. 13 and 14 the ventral side with the mouth parts while Figs. 15 and 16 are the chelicerae of the mite, the cutting organ of the parasite. DISCUSSION One of the unique activity for host attachment for the ticks is “questing”. The first walking appendage has a unique structure called Haller’s organ used for chemical sensing along with terminal claws and pads on the appendages called pulvilli (Ray, 2012). On the stems of grass or edges of leaves the ticks crow in order to come closer to the host. Heat, movement along with carbon dioxide act as stimuli for questing behavior with the help of extended front legs the ticks climb on the potential host (Macnair, 2016). 9 Heckmann Pakistan Journal of Parasitology 65; June 2018 The chemicals released by the host could also be detected by Haller’s organ. The adult ticks have 8 appendages extending from their body (idiosome). Each appendage has a series of joints and terminates with hook-like fingers, called claws, which aid attachment, especially for questing. The Ixode ticks release a number of substances which anticoagulant anti-inflammatory and immunosuppressive, thus the host is unaware of the tick feeding and pathogen establish themselves in the infected host (Francischetti et al., 2009 and John and Ryan, 2017). There have been many studies on the saliva of ticks and the anticoagulants (Jordan et al., 1990; Simo et al., 2017; Kazimirova, 2008 and Bowman et al., 1997). For attachment to a new host, ticks have many anatomical modifications as well as the questing activity. The tip of each appendage has well-developed claws and a pulvillus. Note on Figs. 11A and 11B the pulvillus is well-developed at the top of the first appendage in relation to the same structure on the second appendage (Fig. 12). This structure along with the Haller’s organ can be used for chemical sensing of potential hosts. The pulvilli also have adhesive properties with a spine (empodium) extending between them, there are two pulvilli on each appendage. Pulvilli are lobes or pads that are located between the tarsal claws of many insects (Niederegger et al., 2002 and Sukontason et al., 2006). The pads have adhesive properties, including the use of an adhesive fluid, and this helps the insect or tick stick to the surface on which it is standing. Between the pulvilli there is often a spine called the empodium. The appendages of Arthropods that become ectoparasites have unique modifications on the terminal end of the walking leg for attachment.