Molecular Characterization of Haemaphysalis Species and a Molecular Genetic Key for the Identification of Haemaphysalis of North America

Haemaphysalis longicornis (Acari: Ixodidae), the Asian longhorned tick, is native to East Asia, but has become established in Australia and New Zealand, and more recently in the United States. In North America, there are other native Haemaphysalis species that share similar morphological characteristics and can be difficult to identify if the specimen is damaged. The goal of this study was to develop a cost-effective and rapid molecular diagnostic assay to differentiate between exotic and native Haemaphysalis species to aid in ongoing surveillance of H. longicornis within the United States and help prevent misidentification. We demonstrated that restriction fragment length polymorphisms (RFLPs) targeting the 16S ribosomal RNA and the cytochrome c oxidase subunit I (COI) can be used to differentiate H. longicornis from the other Haemaphysalis species found in North America. Furthermore, we show that this RFLP assay can be applied to Haemaphysalis species endemic to other regions of the world for the rapid identification of damaged specimens. The work presented in this study can serve as the foundation for region specific PCR-RFLP keys for Haemaphysalis and other tick species and can be further applied to other morphometrically challenging taxa

Infestation of introduced raccoons (Procyon lotor) with indigenous ixodid ticks on the Miura Peninsula, Kanagawa Prefecture, Japan

Since the raccoon (Procyon lotor) was introduced to Japan, studies have established that they are infested with native Japanese tick species. However, the quantity of ticks infesting raccoons is unknown. We conducted a survey of ticks on invasive raccoons captured on the Miura Peninsula, Kanagawa Prefecture, Japan, from April 2015 through June 2016 to determine the species of ticks and to quantify the intensity of tick infestation in order to obtain basal information related to the ecology of host–parasite relationships among indigenous tick species and an alien mammalian species. We collected and identified 15,931 ticks of two genera and six species, namely, Haemaphysalis flava, H. megaspinosa, H. longicornis, H. japonica, Ixodes ovatus, and I. tanuki, from 100 out of 115 raccoons. The dominant tick species was H. flava (96.8%) and individuals were mainly adults. Seasonal patterns of infestation intensity of adults and nymphs peaked in the autumn and winter and decreasing in the late spring and summer, May to August, while larvae peaked in August. Our results indicated that host–parasite relationships between invasive raccoons and Japanese tick species, especially H. flava, were established in Kanagawa Prefecture. The ticks infest invasive raccoons for their blood-meal and also for overwintering. The results of this study extend our understanding of the ecology of tick-borne diseases. Keywords: Invasive alien species, Haemaphysalis flava, Intensity of infestation, Procyon lotor, Japa

Quantifying the direct and indirect effects of sika deer (Cervus nippon) on the prevalence of infection with Rickettsia in questing Haemaphysalis megaspinosa: A field experimental study

Sika deer (Cervus nippon) are important hosts for all life stages of Haemaphysalis megaspinosa, a suspected Rickettsia vector. Because some Rickettsia are unlikely to be amplified by deer in Japan, the presence of deer may decrease the prevalence of Rickettsia infection in questing H. megaspinosa. As sika deer decrease vegetation cover and height and thereby indirectly cause changes in the abundance of other hosts, including reservoirs of Rickettsia, the prevalence of Rickettsia infection in questing ticks can also change. We investigated these possible effects of deer on the prevalence of infection with Rickettsia in questing ticks in a field experiment in which deer density was manipulated at three fenced sites: a deer enclosure (Deer-enclosed site); a deer enclosure where deer had been present until 2015 and only indirect effects remained (Indirect effect site); and a deer exclosure in place since 2004 (Deer-exclosed site). Density of questing nymphs and the prevalence of infection with Rickettsia sp. 1 in questing nymphs at each site were compared from 2018 to 2020. The nymph density at the Deer-exclosed site did not significantly differ from that at the Indirect effect site, suggesting that the deer herbivory did not affect the nymph density by reducing vegetation and increasing the abundance of other host mammals. However, the prevalence of infection with Rickettsia sp. 1 in questing nymphs was higher at the Deer-exclosed site than at the Deer-enclosed site, possibly because ticks utilized alternative hosts when deer were absent. The difference in Rickettsia sp. 1 prevalence between the Indirect effect and Deer-exclosed sites was comparable to that between the Indirect effect and Deer-enclosed sites, indicating that the indirect effects of deer were as strong as the direct effects. Examining the indirect effects of ecosystem engineers in the study of tick-borne diseases may be more important than previously recognized

Mapping the Potential Distribution of Ticks in the Western Kanto Region, Japan: Predictions Based on Land-Use, Climate, and Wildlife

Background: Tick distributions have changed rapidly with changes in human activity, land-use patterns, climate, and wildlife distributions over the last few decades. Methods: To estimate potential distributions of ticks, we conducted a tick survey at 134 locations in western Kanto, Japan. We estimated the potential distributions of six tick species (Amblyomma testudinarium Koch, 1844; Haemaphysalis flava Neumann, 1897; Haemaphysalis kitaokai Hoogstraal, 1969; Haemaphysalis longicornis Neumann, 1901; Haemaphysalis megaspinosa Saito, 1969; and Ixodes ovatus Neumann, 1899) using MaxEnt modeling based on climate patterns, land-use patterns, and the distributions of five common wildlife species: sika deer (Cervus nippon Temminck, 1838), wild boar (Sus scrofa Linnaeus, 1758), raccoon (Procyon lotor Linnaeus, 1758), Japanese raccoon dog (Nyctereutes procyonoides Gray, 1834), and masked palm civet (Paguma larvata C.E.H. Smith, 1827)). Results: We collected 24,546 individuals of four genera and 16 tick species. Our models indicated that forest connectivity contributed to the distributions of six tick species and that raccoon distribution contributed to five tick species. Other than that, sika deer distribution contributed to H. kitaokai, and wild boar distribution, bamboo forest, and warm winter climate contributed specifically to A. testudinarium. Conclusions: Based on these results, the dispersal of some tick species toward residential areas and expanded distributions can be explained by the distribution of raccoons and by forest connectivity

Body Length and Craniometrics of Non-Native Raccoons in Two Regions in Middle Japan during Early Invasion Stages

Although the genetic distribution of introduced raccoons (Procyon lotor) in recent years is well known, few studies have examined their morphometrics, especially the relationships between sex and age in the introduced populations. The aim of this study was to describe the morphological characteristics of raccoons from parts of eastern and western Japan during their early invasion stages, focusing on the relationships between body length and the principal component of craniometrics, with region, sex, age class, body length, and body mass index using a regression model. The body length increased more in males than females and in the older age class, supporting the association with intrasexual selection and competition for food resources. Positive relationships for body length and body mass index were found in craniometric analyses, particularly regarding cranial size components, in addition to age class for both sexes, while cranial size also differed between regions for females. The relationship between body length and craniometrics was inconsistent with that of subspecies originating in North America. Given the sympatric distribution of haplotypes of multiple subspecies without reproductive-isolating barriers in North America and in several introduced areas, hybridization must have occurred prior to the introduction or naturalization of this species