Aversion of the invasive Asian longhorned tick to the white-footed mouse,the dominant reservoir of tick-borne pathogens in the U.S.A.

The Asian longhorned tick (Haemaphysalis longicornis) was reported for the first time in the U.S.A. in 2017 and has now spread across 12 states. The potential of this invasive tick vector to transmit pathogens will be determined through its association to hosts, such as the white-footed mouse (Peromyscus leucopus), which is the primary reservoir for the causative agent of Lyme disease (Borrelia burgdorferi) and other zoonotic pathogens. Larval H. longicornis were placed on P. leucopus; 65% of the larvae (n = 40) moved off the host within a short period of time, and none engorged. By contrast, larval blacklegged ticks (Ixodes scapularis) did not move from where they were placed in the ear of the mouse. A laboratory behavioural assay was then conducted to assess the interaction of H. longicornis with the hair of potential mammalian host species in the U.S.A. H. longicornis larvae were significantly less likely to enter the hair zone of P. leucopus and humans compared to the hair of domestic cats, domestic dogs and white-tailed deer. This study identifies a tick–host interaction behaviour, which can be quantified in a laboratory assay to predict tick–host associations and provides insights into how ticks select a host.     

Ticks Collected from Wild and Domestic Animals and Natural Habitats in the Republic of Korea

Ticks were collected from 35 animals from 5 provinces and 3 metropolitan cities during 2012. Ticks also were collected by tick drag from 4 sites in Gyeonggi-do (2) and Jeollabuk-do (2) Provinces. A total of 612 ticks belonging to 6 species and 3 genera were collected from mammals and a bird (n=573) and by tick drag (n=39). Haemaphyalis longicornis (n=434) was the most commonly collected tick, followed by H. flava (158), Ixodes nipponensis (11), Amblyomma testudinarium (7), H. japonica (1), and H. formosensis (1). H. longicornis and H. flava were collected from all animal hosts examined. For animal hosts (n>1), the highest Tick Index (TI) was observed for domestic dogs (29.6), followed by Siberian roe deer (17.4), water deer (14.4), and raccoon dogs (1.3). A total of 402 H. longicornis (adults 86, 21.4%; nymphs 160, 39.8%; larvae 156, 38.9%) were collected from wild and domestic animals. A total of 158 H. flava (n=158) were collected from wild and domestic animals and 1 ring-necked pheasant, with a higher proportion of adults (103, 65.2%), while nymphs and larvae only accounted for 12.7% (20) and 22.2% (35), respectively. Only 7 A. testudinarium were collected from the wild boar (6 adults) and Eurasian badger (1 nymph), while only 5 I. nipponensis were collected from the water deer (4 adults) and a raccoon dog (1 adult). One adult female H. formosensis was first collected from vegetation by tick drag from Mara Island, Seogwipo-si, Jeju-do Province.     

孤雌生殖长角血蜱的哈氏器超微结构与发育

为阐明长角血蜱Haemaphysalis longicornis孤雌生殖种群的哈氏器结构及发育特征, 用扫描电镜对其各虫期哈氏器进行了观察, 分析了血餐对哈氏器发育的影响。结果表明： 该种群幼蜱、 若蜱和成蜱哈氏器形态结构基本相同, 均由前窝和后囊构成。幼蜱前窝感毛6根, 位于同一基盘; 若蜱和成蜱哈氏器相似, 前窝感毛7根, 其中1根孔毛位于外侧基盘, 另6根感毛位于内侧基盘。各虫期饱血后哈氏器大小均比饥饿状态下显著增大（P<0.05）。幼蜱前窝与后囊面积比值与若蜱相比无显著差异（P>0.05）, 若蜱前窝与后囊面积比值与成蜱相比差异显著（P<0.05）。各虫期哈氏器均在发育, 且血餐对哈氏器发育有重要影响。幼蜱至若蜱期哈氏器前窝与后囊的发育速度相似, 若蜱至成蜱期哈氏器前窝发育快于后囊。本研究结果在一定程度上揭示了孤雌生殖长角血蜱的哈氏器发育规律。     

Seasonal Distribution of Ticks in Four Habitats near the Demilitarized Zone,Gyeonggi-do (Province), Republic of Korea

This study describes the seasonal distribution of larvae, nymph, and adult life stages for 3 species of ixodid ticks collected by tick drag and sweep methods from various habitats in the Republic of Korea (ROK). Grasses less than 0.5 m in height, including herbaceous and crawling vegetation, and deciduous, conifer, and mixed forests with abundant leaf/needle litter were surveyed at United States (US) and ROK operated military training sites and privately owned lands near the demilitarized zone from April-October, 2004 and 2005. Haemaphysalis longicornis Neumann adults and nymphs were more frequently collected from April-August, while those of Haemaphysalis flava Neumann and Ixodes nipponensis Kitaoka and Saito were collected more frequently from April-July and again during October. H. longicornis was the most frequently collected tick in grass habitats (98.9%), while H. flava was more frequently collected in deciduous (60.2%) and conifer (57.4%) forest habitats. While more H. flava (54.1%) were collected in mixed forest habitats than H. longicornis (35.2%), the differences were not significant. I. nipponensis was more frequently collected from conifer (mean 8.8) compared to deciduous (3.2) and mixed (2.4) forests.     

Association of the invasive Haemaphysalis longicornis tick with vertebrate hosts,other native tick vectors,and tick-borne pathogens in New York City,USA

Haemaphysalis longicornis, the Asian longhorned tick, is an invasive ixodid tick that has rapidly spread across the northeastern and southeastern regions of the United States since first reported in 2017. The emergence of H. longicornis presents a potential threat for livestock, wildlife, and human health as the host associations and vector potential of this invasive pest in the United States are poorly understood. Previous field data from the United States has shown that H. longicornis was not associated with natural populations of small mammals or birds, but they show a preference for medium sized mammals in laboratory experiments. Therefore, medium and large sized mammals were sampled on Staten Island, New York, United States, to determine H. longicornis host associations and vector potential for a range of human and veterinary pathogens. A total of 97 hosts were sampled and five species of tick (Amblyomma americanum, Dermacentor variabilis, H. longicornis, Ixodes scapularis, Ixodes cookei) were found feeding concurrently on these hosts. Haemaphysalis longicornis was found in the highest proportions compared with other native tick species on raccoons (55.4%), Virginia opossums (28.9%), and white-tailed deer (11.5%). Tissue, blood, and engorged larvae were tested for 17 different pathogens using a nanoscale PCR platform. Infection with five pathogens (Borrelia burgdorferi, Anaplasma phagocytophilum, Rickettsia spp., Mycoplasma haemocanis, and Bartonella spp.) was detected in host samples, but no pathogens were found in any larval samples. These results suggest that although large and medium sized mammals feed large numbers of H. longicornis ticks in the environment, there is presently a low potential for H. longicornis to acquire pathogens from these wildlife hosts.     

Inheritance of weight in Rhipicephalus appendiculatus ticks (Acari: Ixodidae) in the laboratory

A selection of the 10% lightest and 10% heaviest males and females of a population of individually weighed Rhipicephalus appendiculatus Neumann adults was made in two experiments. The offspring of homologous pairs were followed until the next adult stage (light x light, control x control and heavy x heavy). The engorged nymphal weights, unfed adult weights, engorged female weights of the parents, egg mass weights, egg weights, larval scutal lengths, engorged larval weights, unfed nymphal weights, engorged nymphal weights and adult weights of the progeny were determined. No significant differences could be demonstrated between the two lines for egg weight, larval scutal length, engorged larval weight and unfed nymphal weight. Significant differences were found between the egg masses, engorged nymphal weights and adult weights of the two lines. The heritability coefficients of body weight determined from adult to adult were 0.14 and 0.10, respectively, during the first and second experiments. Considering females and males separately, the coefficients were 0.10 and 0.18 during the frist experiment and 0.12 and 0.09 during the repeat experiment respectively.     

The life cycle of Haemaphysalis qinghaiensis (Acari: Ixodidae) ticks under laboratory conditions

The developmental stages in the life cycle of Haemaphysalis qinghaiensis were investigated under laboratory conditions. The larval, nymphal and adult ticks were fed on sheep at 25–27 °C, 50 % relative humidity (RH) and exposed to daylight. All free-living stages were maintained in an incubator (28 °C with 90 % RH and a 12-h photoperiod). The whole life cycle of H. qinghaiensis was completed in an average of 176 days (range 118–247 days). The average developmental periods were 34.44 days for egg incubation; 5.83, 4.20 and 33.70 days for larval pre-feeding, feeding and pre-molting; and 3.88, 5.30 and 46.50 days for nymphal pre-feeding, feeding and pre-molting. The average times for pre-feeding, feeding, pre-oviposition and oviposition of female adult ticks were 2.60, 11.40, 8.50, and 19.35 days, respectively. The results confirmed the positive correlation between the weight of the engorged female and the egg mass laid (r = 0.557, P < 0.05). The reproductive efficiency index and reproductive fitness index in females were 5.49 and 4.98, respectively. Engorged nymphs moulting to females (4.53 ± 0.16 mg) were significantly heavier (P < 0.001) than those moulting to males (3.45 ± 0.19 mg). The overall sex ratio of the adult ticks was 1:1.1 (M:F).     

An Assay for Lateral Line Regeneration in Adult Zebrafish

Due to the clinical importance of hearing and balance disorders in man, model organisms such as the zebrafish have been used to study lateral line development and regeneration. The zebrafish is particularly attractive for such studies because of its rapid development time and its high regenerative capacity. To date, zebrafish studies of lateral line regeneration have mainly utilized fish of the embryonic and larval stages because of the lower number of neuromasts at these stages. This has made quantitative analysis of lateral line regeneration/and or development easier in the earlier developmental stages. Because many zebrafish models of neurological and non-neurological diseases are studied in the adult fish and not in the embryo/larvae, we focused on developing a quantitative lateral line regenerative assay in adult zebrafish so that an assay was available that could be applied to current adult zebrafish disease models. Building on previous studies by Van Trump et al.¹⁷ that described procedures for ablation of hair cells in adult Mexican blind cave fish and zebrafish (Danio rerio), our assay was designed to allow quantitative comparison between control and experimental groups. This was accomplished by developing a regenerative neuromast standard curve based on the percent of neuromast reappearance over a 24 hr time period following gentamicin-induced necrosis of hair cells in a defined region of the lateral line. The assay was also designed to allow extension of the analysis to the individual hair cell level when a higher level of resolution is required.     

Acaricidal potential of active components derived from Alpinia galanga rhizome oils and their derivatives against Haemaphysalis longicornis (Acari: Ixodidae)

Acaricidal activities and acetylcholinesterase (AChE) inhibitory activities were evaluated of active constituents of the essential oil extracted from Alpinia galanga rhizomes cultivated from India and their derivatives against Haemaphysalis longicornis nymphs. In addition, the effect was investigated of active components of A. galanga oil on egg laying of adult females of H. longicornis and egg hatchability. Of the volatile components identified in A. galanga oil, ethyl cinnamate, ethyl methoxycinnamate, and methyl cinnamate at 0.32 mg/cm² resulted in 100% mortality, respectively, indicating that the acaricidal activity of the A. galanga oil against H. longicornis nymphs could be attributed to these compounds. To evaluate the structure–activity relationship between cinnamate derivatives and their acaricidal activities, allyl cinnamate, benzyl cinnamate, isopropyl cinnamate, isobutyl cinnamate, and isoamyl cinnamate were selected. Among cinnamate derivatives tested, allyl cinnamate exhibited the most potent toxicity (LC₅₀?=?0.055 mg/cm²) against H. longicornis nymphs. The allyl cinnamate was also tested for AChE activity in vivo in H. longicornis nymphs and was found to affect the AChE activity. Allyl cinnamate at 10–50 mg/mL inhibited egg laying of adult females of H. longicornis by 10–43%. Egg hatching was suppressed completely by treatment with allyl cinnamate at 50 mg/mL, whereas allyl cinnamate was minimally toxic against non-target earthworms, Eisenia fetida. These results suggest that allyl cinnamate can be used as an active ingredient for the development of eco-friendly tick acaricides against H. longicornis, a vector for Sever fever with thrombocytopenia syndrome (SFTS) virus.

     

No Detection of Severe Fever with Thrombocytopenia Syndrome Virus from Ixodid Ticks Collected in Seoul

Larvae, nymphs, and adult stages of 3 species of ixodid ticks were collected by tick drag methods in Seoul during June-October 2013, and their infection status with severe fever with thrombocytopenia syndrome (SFTS) virus was examined using RT-PCR. During the period, 732 Haemaphysalis longicornis, 62 Haemaphysalis flava, and 2 Ixodes nipponensis specimens were collected. Among the specimens of H. longicornis, the number of female adults, male adults, nymphs, and larvae were 53, 11, 240, and 446, respectively. Ticks were grouped into 63 pools according to the collection site, species, and developmental stage, and assayed for SFTS virus. None of the pools of ticks were found to be positive for SFTS virus gene.     

The temperature and humidity preferences of Haemaphysalis longicornis, Ixodes Holocyclus and Rhipicephalus sanguineus (Ixodidae): studies on eggs

The eggs of Haemaphysalis longicornis, Ixodes holocydus and Rhipicephalus sanguineus were exposed to a range of temperatures and humidities to compare the temperature and humidity requirements of ticks from climatically dissimilar habitats. The response to changes in humidity was also studied with H. longicornis. Hatching in I. holocydus eggs occurred from 18 to 28°C and at a saturation deficit of 2 mm Hg or less. The eggs of R. sanguineus hatched between 18 and 38°C and tolerated saturation deficits up to 20 mm Hg. The eggs of H. longicornis hatched between 18 and 35°C and at saturation deficits up to 8 mm Hg. When eggs of H. longicornis were exposed to moderately dry conditions for different periods of time and then transferred to moist conditions, incubation periods and mortality were reduced in comparison with eggs kept continuously under dry conditions. On the other hand, mortality of H. longicornis eggs was reduced in moderately dry conditions, provided that they had been exposed to a moist environment for about 10 days previously. Weight loss of eggs in dry air and the rate of development of eggs were examined and are discussed. The eggs of I. holocylus lost weight very rapidly in dry air, whereas weight loss from R. sanguineus eggs was much slower. Weight loss from H. longicornis eggs fell between these two rates. The rate of development was fastest in R. sanguineus and slowest in I. holocydus.     

Life cycle and host specificity of Amblyomma triste (Acari: Ixodidae) under laboratory conditions

We report biological data of two generations of Amblyomma triste in laboratory and compared the suitability of different host species. Infestations by larval and nymphal stages were performed on guinea pigs (Cavia porcellus), chickens (Gallus gallus), rats (Rattus norvegicus), rabbits (Oryctolagus cuniculus), wild mice (Calomys callosus), dogs (Canis familiaris) and capybaras (Hydrochaeris hydrochaeris). Infestations by adult ticks were performed on dogs, capybaras and rabbits. Tick developmental periods were observed in an incubator at 27 °C and RH 90%. Guinea pigs were the most suitable hosts for larvae and nymphs, followed by chickens. The remaining host species were less suitable for immature ticks as fewer engorged ticks were recovered from them. Mean larval feeding periods varied from 3.8 to 4.7 d between different host species. Mean larval premolt periods ranged from 8.9 to 10.4 d. Nymphal mean feeding periods varied from 4.2 to 6.2 d for ticks fed on different host species. Premolt period of male nymphs (mean: 15.4 d) was significantly longer than that of female nymphs (14.7 d). Female nymphs were significantly heavier than male nymphs. The overall sex ratio of the adult ticks emerged from nymphs was 0.9:1 (M:F). Capybaras were the most suitable host for the tick adult stage as significantly more engorged females were recovered from them and these females were significantly heavier than those recovered from dogs or rabbits. The life cycle of A. triste in laboratory could be completed in an average period of 155 d. The potential role of guinea pigs, birds and capybaras, as hosts for A. triste in nature, is discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

Evaluation of the acaricidal activity of 63 commercialized pesticides against Haemaphysalis longicornis (Acari: Ixodidae)

The acaricidal activity and residual effects of 63 commercialized pesticides against the Cheongju strain of Haemaphysalis longicornis in Korea were examined. Twenty‐two pesticides (4 carbamates, 5 organophosphates, 10 pyrethroids, 1 amitraz, 1 diamide, and 1 unclassified pesticide) caused greater than 80% mortality in H. longicornis adults under laboratory conditions using the spray method. These 22 pesticides were used to treat grassland under field conditions for investigation of the residual effects, and 100% mortality in H. longicornis adults was observed with all the carbamates (carbosulfan, benfuracarb, fenobucarb, and carbaryl) and γ‐cyhalothrin after 3 days of pesticide treatment. γ‐Cyhalothrin exhibited 56.7% mortality after 10 days of treatment, which was the longest residual effect of treated grass on H. longicornis adults. With regard to the residual effects on H. longicornis under field conditions, most of the 22 pesticides exhibited higher mortality in nymphs than in adults. In particular, benfuracarb exhibited 96.7% acaricidal activity until 10 days after treatment. These results indicate that carbamates are highly likely to be available under field conditions and, based on this preliminary data, could be used for the control of H. longicornis adults and nymphs.     

Detachment of hard ticks (Acari: Ixodidae) from hunted sika deer (Cervus nippon)

Ixodid ticks were collected from 13 sika deer, Cervus n. nippon, shot in the Boso Peninsula in central Japan from late February to early March 1999. Haemaphysalis megaspinosa was the most abundant species of the adults collected, although Haemaphysalis longicornis, H. flava, H. kitaokai, H. cornigera, Ixodes ovatus, and Amblyomma testudinarium were also collected. Males were more abundant than females for H. longicornis, H. megaspinosa, H. flava, and H. kitaokai. Ticks that had inserted their hypostome into its host skin (designated attached) were distinguished from those that were detached and on the host’s surface. A greater fraction of males than females of all four species were detached. Females were classified in three feeding stages (engorged, partially engorged, and unfed). More H. longicornis and H. megaspinosa unfed female ticks than engorged and partially-engorged female ticks were collected detached. Our results indicated that H. megaspinosa, H. longicornis, H. flava, and H. kitaokai male ticks detached sooner than female ticks after their host died.     

The differential mortality at various life stages of the greenhouse whitefly,Trialeurodes vaporariorum (Homoptera: Aleyrodidae), by infection with the fungus Aschersonia aleyrodis (Deuteromycotina: Coelomycetes)

The parasitoid Encarsia formosa is commonly applied to control the greenhouse whitefly Trialeurodes vaporariorum in glasshouse tomatoes and cucumbers. Nevertheless, in some cases the control capacity of this natural enemy is insufficient and an additional selective pest-suppressing agent is desirable. The entomopathogenic fungus Aschersonia aleyrodis was applied to cucumber plants carrying whiteflies in different developmental stages. After spraying each leaf with 2 ml of spore suspension (4 × 10⁶ spores/ml) the plants were kept at 100% RH for 24 hr; thereafter the humidity was lowered to 70% RH at 20°C and the photoperiod was 16 hr. Treated eggs did not become infected, but larvae that hatched from these eggs and settled on the treated abaxial leaf surface were infected at the same rate and to the same degree as treated first instar larvae. This suggests that the spores persist for at least 7 days. The final percentages of infection over all instars when treated as young eggs, old eggs, and first larval instars were 94, 93, and 90%, respectively. The final percentages of infection when treated as third and fourth larval instars and prepupae were 76, 28, and 12%, respectively. The older instars were less susceptible and adults were seldom infected by the fungus. Several applications of A. aleyrodis as a microbial insecticide are needed to achieve sufficient control of whitefly populations in glasshouses.     

Patterns of Molecular Variation. II. Associations of Electrophoretic Mobility and Larval Substrate within Species of the DROSOPHILA MULLERI Complex

Electromorphic variation among populations of Drosophila mojavensis, D. arizonensis and D. longicornis was examined for seven genetic loci. The average electrophoretic mobility for a population was used as the metric. D. mojavensis and D. arizonensis use larval substrates in different parts of their geographic ranges, while D. longicornis is more narrowly restricted to different species of the cactus Opuntia in different localities. There is marked electromorphic variation among populations of either D. mojavensis or D. arizonensis, and the bulk of this variation is accounted for by differences in laval substrate. There is somewhat less variation among populations of D. longicornis, and only a moderate portion of this is accounted for by larval substrate differences. There appears to be an association between the taxonomic diversity of the larval substrates and the electromorphic diversity of the Drosophila populations utilizing those substrates. Evidence is reviewed that suggests physiological mechanisms for these possibly adaptive associations.     

The development of oocytes in the ovary of a parthenogenetic tick, Haemaphysalis longicornis

Haemaphysalis longicornis is an important vector of various pathogens in domestic animals and humans. The tick is a unique species with bisexual and parthenogenetic races. Although mating induces oocyte development, it is possible in the parthenogenetic race to complete oogenesis without copulation. Here we examined the developmental process of oocytes from unfed to the oviposition period in parthenogenetic H. longicornis. We classified the developmental stages of oocytes into five stages: stage I, germinal vesicle occupies more than half of the cytoplasm; stage II, germinal vesicle occupies less than half of the cytoplasm; stage III, germinal vesicle migrates from the center in the oocyte to the vicinity of the pedicel cells; stage IV, the cytoplasm is filled with yolk granules of various sizes; stage V, the cytoplasm is occupied by large yolk granules. Oocytes at the unfed period were undeveloped and classified as stage I. Stage I and II oocytes were observed at the rapid feeding period, indicating that oocyte development began after the initiation of blood feeding. All developmental stages of oocytes were observed at the pre-oviposition period. At 10?days after the beginning of the oviposition period, the ratios of stage I and II oocytes were higher than those of the previous period, suggesting that the ovarian development and activity may be continuing. Based on these findings, we propose classification criteria for the oocyte development in the parthenogenetic H. longicornis. The criteria will be useful for understanding the mechanisms of tick reproduction and transovarial transmission of pathogens.