Survey of Ticks on Reptilian Hosts on the Island of Dominica, West Indies. Texas A&M University. Study Abroad Dominica 2013.

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Survey of s on Reptilian s on the Island of Dominica, West Indies Texas A&M University Study Abroad Dominica 2013 Emily Crews Dr. Jim Woolley Dr. Thomas Lacher

Crews 2 Abstract A survey of ticks throughout various locations on Dominica was taken to explore the distribution and frequency of different tick species on several animal hosts. This survey focused on ticks extracted from reptilian hosts, namely Ameiva (or the Dominican Ground Lizard) and Boa nebulosa (or the Boa Constrictor). Results indicated that ticks of the genus appear most frequently, specifically on large reptiles. Small reptiles such as Anolis oculatus (the Dominican Anole), Thecadactylus rapicauda (Turnip-Tailed Gecko), Hemidactylus mabouia (House Gecko), Liophis juliae (Julia s Ground Snake), and Alsophis antillensis (Antilles Racer) yielded no ticks, however several mites were observed. Introduction s, members of the class Arachnidae, are eight-legged arthropods that survive through parasitism of a host animal. A tick invades its host by inserting its needle-like, barbed hypostome into the host s skin to take a bloodmeal (CDC.gov, 2013). The tick s ability to take a bloodmeal is crucial to its life cycle, as it will remain in the larval stage until it has successfully fed. Once engorged with blood, the tick drops from the host, molts, and seeks a new host (Palacios et al, 2009). Potential hosts include mammals, reptiles, birds, and amphibians (CDC.gov, 2013). The life cycle of a tick contains four stages: egg, larva, nymph, and adult. This cycle can potentially last for three years. After hatching from the egg, a tick will seek out a host in its sixlegged larval form. Once it successfully attaches to a host and engorges itself with blood, it detaches from its host, molts (developing a fourth pair of legs), and seeks a host for its life as a nymph. After doing so, it will drop and molt into its larger, more formidable adult stage and seek a final host. The adult female will engorge herself completely on this new host s blood, while the

Crews 3 smaller adult male will attach to feed, but also to mate with the female. The male will fertilize the female as she takes her bloodmeal, and finally she will drop to lay her hundreds to thousands of eggs (Wilson, 2007). Thus, the cycle begins anew. s can vector several dangerous diseases, specifically Babesiosis on the island of Dominica. Babesiosis infects both Creole and Holstein cattle on the island, is potentially fatal, and can infect humans as well (Palacios et al, 2009). In regard to reptiles, studies have shown that certain lizard species in the Western United States are crucial to the survival of ticks that carry Lyme Disease (Parry, 2011). Lyme Disease has yet to show itself significantly on Dominica, but this is appropriate to consider should the disease migrate. These diseases are transferred from tick to host through tick salivation during feeding, or the tick s secretion of a cement-like fluid that makes removal of the tick more difficult. These fluids can potentially contain pathogens that will infect the host with the disease, be it Babesiosis, Lyme Disease, Rocky Mountain Fever, or otherwise. The tick has usually received the pathogen by feeding on the blood of a previous host, thus spreading the disease among species with potentially no other contact (Wilson, 2007). The purpose of this survey was to determine how commonly ticks attach to reptiles on the island, and what species specifically seem to favor reptilian hosts. Several anatomical structures aid in the identification of ticks, such as the capitulum (which contains a tick s mouthparts, including the hypostome), the body or idiosoma, the scutum, the eyes (which may or may not be present), spines on the base of the legs or coxae, and the anal groove (Strickland & Diamant, 1976). This experiment used these identifying features to determine the species of the collected ticks in order to pinpoint the tick species that frequent reptiles for bloodmeals. The results of this

Crews 4 experiment will serve as informational data for future surveys, as well as a measurement of Lyme s potential to spread should it ever infest the island. Materials and Methods Capture of Ameiva and Subsequent Extraction To conduct the survey, several locations on the island were scoured for tick-infested hosts, including Batalie, Cabrits National Park, and Springfield Research Center. Ameiva on Batalie were discovered to be particularly susceptible to ticks, thus a method of capture was developed to obtain samples from these quick animals. A group of fellow students aided the author in herding scurrying Ameiva into an old fish net grounded by rocks. The net was thrown over the lizard and students proceeded to restrain the animal. The surveying student then used Watchmakers Forceps. 5 to extract ticks from their reptilian host whilst other students restrained the animal by hand. s were removed at the base of the head and carefully twisted, hopefully to remove the entire tick and avoid breaking any mouthparts. This method of capture was attempted for all three Ameiva hosts, but two of the three were caught by hand. Once ticks were pulled from the animal, they were placed in a 2 ml plastic vile filled with 95% ethyl alcohol. The alcohol s purpose was to kill the ticks, thus making them easier to observe and less dangerous to the student handling them. Vials were placed in numerical order according to which host they were taken from and labeled with the host s species. After all visible ticks were extracted, the host animal was observed to identify any injuries it may have sustained. If necessary, injuries were washed with ninety-five percent ethyl alcohol to prevent infection. The reptile was then released to the wild.

Crews 5 Capture of Boa nebulosa and Subsequent Extraction The same locations searched for Ameiva were also explored for other hosts. One host that frequented the scene was Boa nebulosa. Boa nebulosa was caught more easily due to its slow-paced slither. However, more caution was used to catch Boa nebulosa because of its behavior. The method to catch boas consisted of pursuing them, then quickly placing a hand behind their head and grabbing the tail with a free hand. Some boas, due to their large size, required more than one person to restrain. The dorsal and ventral scales of each boa were manipulated to reveal ticks that had possibly attached beneath them. Beyond this step, the process closely mirrored the extraction method used for Ameiva hosts. Examination of Specimens, Tentative Identification, and Photography After being harvested, ticks were returned to the lab and kept at room temperature until examination, usually within twenty-four hours of capture. Each tick was examined under a Leica EZ4 microscope at varying degrees of magnification. A petri dish was filled with roughly two milliliters of alcohol and the ticks from one host were carefully placed side-by-side. Both sides of the ticks were observed for characteristic anatomical structure, such as the shape of the dorsal shield, length of palps versus hypostome, presence of eyes, and coloration patterning. Particular attention was paid to identification of the genus as these ticks frequent the Caribbean (Texas A&M University, 2011). Distinctive characteristics were taken note of. This process was repeated for each host s ticks until all ticks had been tentatively identified.

Crews 6 After examination had taken place, the student photographed ticks of interest or superior quality with the help of the professor. A Nikon D300 camera and a Zeiss 100mm macro lens were used with considerable lens extension. Several remote flash aids were used to illuminate the ticks due to their miniature size. s were left in a small petri dish and placed on white copy paper for photographic purposes. These images were then emailed to Dr. Pete Teel of Texas A&M University for confirmation of identity. The ticks were carefully replaced into their respective vials using the watchmaker s forceps after being successfully photographed. After each use, the watchmaker s forceps were cleansed with 95% ethyl alcohol. Voucher specimens for tick collections were labeled and stored in 95% ethyl alcohol and deposited in the insect collection of ATREC, Springfield. Results The two tick species discovered were tentatively identified as americanum (or Lonestar tick) and (or Cayenne tick). These names are used in this report pending confirmation by tick specialists. Table 1 shows the results of all tick collections. Figures 1-6 show photographed specimens.

Crews 7 Date of Collection of Collection 21/5/2013 Springfield Research Station 24/5/2013 Batalie 24/5/2013 Batalie 24/5/2013 Batalie 24/5/2013 Batalie 24/5/2013 Batalie #1 Boa #2 Ameiva #2 Ameiva #2 Ameiva #2 Ameiva #2 Ameiva (Tentative) #1 Americanum (Lonestar ) #2 ) #3 ) #4 ) #5 ) #6 ) on Behind Left Eye Ventral Ventral Ventral Front Left Leg Back Right Leg tes Adult Female, Substantially Large White Spot on Scutum Photo Taken Y/N (dorsal and ventral) (dorsal) (ventral)

Crews 8 Date of Collection of Collection #3 Ameiva #3 Ameiva #3 Ameiva #3 Ameiva #3 Ameiva #4 Ameiva #4 Ameiva #4 Ameiva #5 Boa (Tentative) #7 ) #8 ) #9 ) #10 ) #11 ) #12 ) #13 ) #14 ) #15 ) on Ventral Ventral Ventral Right Hind Leg tes Male, Male, Male, Adult Female, Partially Engorged Male, Male, Male, Male, Photo Taken Y/N (dorsal) (ventral) (dorsal and ventral) (dorsal and ventral)

Crews 9 Date of Collection of Collection #5 Boa #5 Boa #5 Boa #5 Boa #5 Boa #5 Boa (Tentative) #16 ) #17 ) #18 ) #19 ) #20 ) #21 ) on Ventral tes, Odd Off- White Protrusion from Photo Taken Y/N (dorsal and ventral) (dorsal and ventral)

Crews 10 Date of Collection of Collection (Tentative) on tes Photo Taken Y/N #22 Unidentified #23 Unidentified #24 Unidentified #25 Unidentified #26 ) #27 Unidentified #28 Unidentified #29 Unidentified #30 Unidentified Male, Table 1, location and reptile hosts of all ticks collected.

Crews 11 Figures 1-2. americanum. 1 View. 2 Ventral View. Figures 3-4.. 3 View. 4 Ventral View. Figure 5-6. (nymph). 5 View. 6 Ventral View.

Crews 12 Discussion As the data above exemplifies, ticks of the genus heavily predominated the ticks gathered, particularly. Males of the species are overwhelmingly present. Photographs will be sent to an expert at Texas A&M University for confirmation of this tentative identification. Larval ticks proved very difficult to identify, although it is predicted that these are also of the genus Based on the sampling taken during this survey, the genus has been identified as the prominent tick to plague reptiles on the island of Dominica, particularly in Ameiva and Boa nebulosa. This genus was pinpointed by using anatomical characteristics to identify the ticks as prospective members of the genus. These ticks had an indistinct anal groove behind the anus, long palps, a non-hexagonal dorsal shield, eyes subtly present, an ornate scutum (dorsal shield), well-developed festoons, and males lacking distinctive adanal shields. All are identification characteristics of ticks (Strickland & Diamant, 1976). Photographic references appear to confirm the speculative species due to coloration and key coloration patterning. According to collected data, a correlation between tick maturity and reptile size was observed. The smaller host, Ameiva, tended to be infested with small adults or even nymphs, while the larger Boa nebulosa was favored by larger adult ticks. This proved true of the first two Boa nebulosa hosts captured. A significant outlier was present in the tick larvae that were collected from host number six, a seven-foot long boa constrictor. A wider sampling would need to be taken to determine whether this correlation was simply a coincidence from collection

Crews 13 (thus proving the outlier a normal occurrence) or if mature ticks do prefer larger animals. This does tend to be true in the United States in regard to mammals (Wilson, 2007). Another curious observation made was that the spines often present on the coxae of do not seem to be present in the collected specimens. It is possible that this characteristic could have been mutilated during extraction from the host, although this seems unlikely. A speculative explanation would be that the shortage of mammals on the island of Dominica has caused the to evolve without these spines that could potentially be used to hook on to fur. A final curious observation is that ticks do not seem to infect relatively small reptiles such as Anoles or Geckos. This could be due to the tightly-spaced scales of these smaller animals, thus making it harder or even impossible for the ticks to imbed their hypostomes into the reptilian flesh. Smaller hosts also do not supply as ample a bloodmeal and do not admit as much carbon dioxide or commotion, both factors that attract a questing (host-seeking) tick (Wilson, 2007). Also, a small host would support fewer ticks than a larger host, limiting reproductive potential. A larger host offers more surface area for males to encounter feeding females. Overall, have shown to be the most dominant genus of tick present on the island s reptilian hosts. These ticks are Cayenne ticks or a very similar species. It would be interesting to explore the absence of spines on coxae further to determine whether in fact they represent another species. Sources of error in this experiment include mutilation of ticks through extraction, thus altering identifying anatomical structures or removing them completely. Other errors could

Crews 14 include overlooking ticks present on animals captured and assumed to be tick-free, or simply overlooking significant ticks on hosts already harvested for specimens. Other issues include the alteration of color after the ticks are stored in ethyl alcohol for extended periods of time. Conclusion s are dangerous to both humans and animals, as they commonly vector diseases that are life-threatening. Their host-dependent life cycle makes them a crucial means for zoonotic diseases to transfer to human life. This survey was designed to determine the variety of ticks present on Dominican reptiles and whether they could potentially vector diseases to livestock or the humans that handle them. Experimental results from reptilian hosts yielded a strong presence of ticks of the genus, a group of ticks known to vector significant diseases such as Rocky Mountain Fever and Equine piroplasmosis (Texas A&M University, 2011). Thus, ticks on reptiles may have significant impacts on the livestock or people of Dominica. Acknowledgements Many thanks to Dr. Jim Woolley and Dr. Thomas Lacher for their endless patience, guidance, and chauffeur services for research purposes. Thanks are also owed for Dr. Woolley s talent for photography and Dr. Lacher s willingness to answer endless requests for scientific names. Thank you, professors!

Crews 15 References Cited "Cayenne." The App for Texas and The Southern Region. Texas A&M University, 2011. Web. 04 June 2013. < http://tickapp.tamu.edu/ticks/cayennetick.php> Palacios, Monica, Kyle Toomey, Lakeisha Shaw, and Lindsey Hranitzky. "A Study of Babesia in Cattle on the Island of Dominica." Dominica 2013. Texas A&M University, Jan. 2013. Web. 4 June 2013. <http://dominica.tamu.edu/student%20projects/dominica%20projects%20pdf%20copy/p alacios_group.pdf> Parry, Wynne. "Without Lizards to Latch Onto, Lyme Disease s Depart."LiveScience.com. TechMediaNetwork.com, 16 Feb. 2011. Web. 04 June 2013. <http://www.livescience.com/12884-fence-lizards-lyme-disease-ticks.html>. Strickland, Robert K., and Gerald Diamant. s of Veterinary Importance. Washington: Animal and Plant Health Inspection Service, U.S. Dept. of Agriculture, 1976. Print. "s." CDC.gov. Centers for Disease Control and Prevention, 10 May 2013. Web. 04 June 2013. <http://www.cdc.gov/ticks/> Wilson, Tracy V. "How s Work." HowStuffWorks. Discovery, 29 Aug. 2007. Web. 04 June 2013. <http://science.howstuffworks.com/zoology/insects-arachnids/tick.htm>