ECDYSTEROIDS IN THE LARVA OF HAEMAPHYSALIS LONGICORNIS NEUMANN (ACARI: IXODIDAE)*

Abstract The ecdysteroids (ECs) concentration and their components in the larvae of Haemaphysalis longicornis were determined by radioimmunoassay (RIA) and high performance liquid chromatography (HPLC). ECs ware found in feeding and engorged larvae and absent in unfed larvae. During feeding and the first 3 days after engorgement, ECs were in low level (less than 14. 35pg EE /larva). On the 5th day, they began to increase sharply (29. 87pg EE /larva) and reached their peak (56. 04pg EE /larva) on the 7th day. Then they declined to low value again (19. 38pg EE /larva). HPLC analysis revealed that the ECs components of the RIA peak could be 20-hydroxyecdysone (20E) and ecdysone (E) on the ratio of 20E: E = 3. 07: 1, which meant 20E was probably the main effective hormone.     

Intraguild predation among Scolothrips longicornis (Thysanoptera: Thripidae), Neoseiulus californicus and Typhlodromus bagdasarjani (Acari: Phytoseiidae) under laboratory conditions

This study was carried out on the ability of predatory thrips Scolothrips longicornis Priesner to feed on 2 phytoseiid species and vice versa. Also the effect of predation of Neoseiulus californicus (McGregor) on Typhlodromus bagdasarjani Wainstein and Arutunjan and vice versa was evaluated. The larvae, prepupae, and pupae of thrips and the eggs, larvae, and protonymphs of phytoseiids were selected as intraguild prey. The intraguild predation (IGP) among S. longicornis and 2 phytoseiid species was unidirectional and in favor of phytoseiids, i.e., S. longicornis was not able to feed on larval stages of 2 phytoseiids. However, N. californicus and T. bagdasarjani fed on the 1st instar larvae (1.39 and 0.80 per day), 2nd instar larvae (0.87 and 0.55 per day), prepupae (0.51 and 0.48 per day), and pupae of thrips (0.51 and 0.49 per day, respectively). Both phytoseiids fed on eggs, larvae, and protonymphal stages of each other. Females of N. californicus consumed more phytoseiid larvae (2.49 per day) than T. bagdasarjani, which consumed 1.08 N. californicus larvae per day. When Tetranychus urticae was presented as an extraguild prey, intensity of IGP between 2 species of phytoseiids and on larval stages of S. longicornis reduced significantly. Therefore, it is concluded that (i) IGP existed among the 3 examined species and lack of feeding of S. longicornis on 2 phytoseiid species can be justified by its feeding type (monophagy), (ii) N. californicus was much more prone to IGP than was T. bagdasarjani.     

Autophagy-related genes from a tick,Haemaphysalis longicornis

Ticks are gorging-fasting organisms; their life cycle is characterized by alternate off-host (starvation) and on-host (meal) conditions. Their generation time is estimated in several years and many ticks spend more than 95% of their life off the host. They seem to have a unique strategy to endure the off-host state for a long period. Thus, we focused on autophagy, which is induced by starvation and is essential for extension of the lifespan, and hypothesized that ticks also have a system of autophagy to overcome the starved condition. Recently, we showed the existence of a homologue of an ATG gene, ATG12, and its expression pattern from nymphal to adult stages in a three-host tick, Haemaphysalis longicornis. The expression level of HlATG12 was downregulated at the beginning of feeding and was highest at 3 months after engorgement. In addition, the HlAtg12 protein was localized to the region around granule-like structures within midgut cells of unfed adults. These results indicate that HlATG12 functions during unfed stages. Here, a potential role of autophagy in unfed ticks is discussed with regard to reports in other animals, such as yeast, mammal, and fruit fly.

Addendum to: Umemiya R, Matsuo T, Hatta T, Sakakibara S, Boldbaatar D, Fujisaki K. Cloning and characterization of an autophagy-related gene, ATG12, from the three-host tick Haemaphysalis longicornis. Insect Biochem Mol Biol 2007; 37:975-84     

Goats,hares, and rabbits as hosts for the New Zealand cattle tick,Haemaphysalis longicornis

Abstract

Feral goats and hares were commonly infested by immature stages of the New Zealand cattle tick, Haemaphysalis longicornis. No explanation could be found for the low prevalence of adult ticks on these hosts. The ears of both host species were almost the exclusive feeding site of the ticks and this may be a consequence of grooming behaviour. Another potential host, the rabbit, was examined but few were found to be infested.

The less restricted range of non-domesticated hosts, together with feeding habits that differ from domestic stock, make them an important additional source of information on the ecology and seasonal pattern of activity of H. longicornis. Also, they are a source of contamination for tick-free pasture, and could possibly maintain the tick population in the absence of sheep and cattle. It is important that their role as alternative hosts be understood and considered in tick-control programmes.     

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.     

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.     

Selective grazing of Temora longicornis in different stages of a Phaeocystis globosa bloom—a mesocosm study

Selective grazing of a calanoid copepod Temora longicornis was measured during different stages of a Phaeocystis globosa bloom, in order to reveal (1) if T. longicornis feeds on single cells and/or colonies of P. globosa in the presence of alternative food sources, (2) if copepod food selection changes during the initiation, maintenance, collapse and decay of a P. globosa bloom and (3) if P. globosa dominated food assemblage provides a good diet for copepod egg production. Our results show low but constant feeding on small colonies of P. globosa, irrespective of the type or concentration of alternative food sources. In contrast, feeding on single cells was never significant, and the total contribution of P. globosa to carbon ingestion of T. longicornis was minor. T. longicornis fed most actively on the decaying colonies, whereas during the peak of the bloom copepods selected against P. globosa. Mostly, T. longicornis fed unselectively on different food particles: before the bloom, the major part of the diet consisted of diatoms, whereas during and after the bloom copepod diet was dominated by dinoflagellates and ciliates. Egg production was highest during the decay of the bloom, coinciding with highest proportional ingestion of heterotrophic organisms, but was not seriously reduced even during the peak of the bloom. We conclude that P. globosa blooms should not threaten survival of copepod populations, but the population recruitment may depend on the type (and concentration) of the dominant heterotrophs present during the blooms. Due to relatively unselective grazing, the impact of T. longicornis to the initiation of a Phaeocystis bloom is considered small, although grazing on decaying colonies may contribute to the faster termination of a bloom.     

QUANTITATIVE VARIATION OF 20-HYDROXYECDYSONE IN THE NYMPH OF DERMACENTOR NIVEUS NEUMANN*

Abstract The variation of 20-hydroxyecdysone (20 E) levels in the unfed, feeding and engorged nymphs of Dermacentor niveus was studied. The results showed that 20 E existed in all stages of nymphal development. There were two peaks. A small peak (2. 34 ng/nymph) appeared in the feeding period (5 days after attachment) which was connected with the fast feeding before dropping from the host, and a big peak (16.17 ng/nymph) in the moulting period (8 days after engorgement). The latter might be correlated with epicuticle deposition. From the 6th day to the 8th day after engorgement, the level of 20 E rose rapidly. Apolysis occurred in this period.     

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.

     

QUANTITATIVE VARIATION OF ECDYSTEROIDS OF IXODID TICKS

Abstract  In order to explore the role of ecdysteroids in development and reproduction of ixodid ticks, we studied the quantitative variation of ecdysteroids in the hemolymph, synganglion, ovary and whole body of the female ixodid ticks, Dermacentor niveus and Haemaphysalis longicornis , before and after engorgement and oviposition by HPLC and RIA. The ecdysteroid content in eggs of these ticks was determined by HPLC. The results indicated that before engorgement the quantitative variation of ecdysteroids in the whole female body was not significant, but their levels increased rapidly after engorgement. The ecdysteroid titer in hemolymph was peaked on the 5th day after engorgement, which was one day prior to oviposition. It may be regarded as a singnal of oviposition. In the synganglion the peak of ecdysteroid level occurred also on the 5th day after engorgement. This is coincident with the secretory activity of neurosecretory cells of synganglion. From the 3rd day after engorgement until oviposition the ecdysteroid level in the ovary increased rapidly. Ecdysteroids were detected in eggs of H. longicornis too. They stem from ovary and accumulated with the process of embryonic development.     

Coexistence of two species of whip spiders (Genus Heterophrynus) in the neotropical rain forest (Arachnida,Amblypygi)

Summary Heterophrynus longicornis Butler and H. alces Pocock are sympatric and syntopic in Serra do Navio in the central part of Amapá, N.-E. Brazil. Both species are of similar sizes during part of their life cycles. Competition is probably reduced by utilization of different hiding places and strata. In H. longicornis, adults are tolerant towards conspecifics. Male and female are usually found close to each other and, under laboratory conditions, courtship behaviour can be observed even when the female is not receptive. H. alces, on the other hand, is very intolerant, and usually only single specimens are found. Under laboratory conditions, cannibalism is frequent. The ecological implications of these differences are discussed.     

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.     

Immunofluorescent detection in the ovary of host antibodies against a secretory ferritin injected into female Haemaphysalis longicornis ticks

Due to the continuous threat of ticks and tick-borne diseases to human and animal health worldwide, and the drawbacks of chemical acaricide application, many researchers are exploring vaccination as an alternative tick control method. Earlier studies have shown that host antibodies can circulate in the ticks, but it has not been confirmed whether these antibodies can be passed on to the eggs. We previously reported that ticks infesting rabbits immunized with a recombinant secretory ferritin of Haemaphysalis longicornis (HlFER2) had reduced egg production and hatching. Here we attempted to detect the presence of antibodies against HlFER2 in the ovary and eggs of female ticks through immunofluorescent visualization. Purified anti-HlFER2 antibodies or rabbit IgG for control was directly injected to engorged female H. longicornis. Ovaries and eggs after oviposition were collected and prepared for an indirect immunofluorescent antibody test. Positive fluorescence was detected in ovaries one day post-injection of anti-HlFER2 antibodies. Through silencing of Hlfer2 gene, we also determined whether the injected antibodies can specifically bind to native HlFER2. Immunofluorescence was observed in the oocytes of dsLuciferase control ticks injected with anti-HlFER2 antibodies, but not in the oocytes of Hlfer2-silenced ticks also injected with anti-HlFER2 antibodies. Our current findings suggest that host antibodies can be passed on to the oocytes, which is significant in formulating a vaccine that can disrupt tick reproduction.     

Artificial immunization of rabbits with Hyalomma dromedarii tick-derived midgut antigen

New Zealand white rabbits were immunized with partly fed Hyalomma dromedarii tick-derived midgut concealed antigens (supernate and pellet fractions) and Freund's complete adjuvant (FCA). The rabbits received three inoculations subcutaneously on days 0, 14 day 21 at a dose rate of 1 mg antigen per animal. The effects of the immunity induced was determined by infesting the rabbits with adult H. dromedarii ticks. In immunized rabbits a significant reduction in tick yield, engorgement weight, oviposition period, egg mass weight and percentage of egg hatchability was found. The gut supernatant antigen fraction induced the best protection in terms of reduced feeding and reproductive performance of the ticks.     

Discrimination between <Emphasis Type="Italic">Haemaphysalis longicornis</Emphasis> and <Emphasis Type="Italic">H. qinghaiensis</Emphasis> based on the partial 16S rDNA and the second internal transcribed spacer (ITS-2)

In the present study, two hard tick species, Haemaphysalis longicornis and H. qinghaiensis from North-western China were characterized genetically by the second internal transcribed spacer (ITS-2) of nuclear ribosomal DNA and partial 16S rDNA. Based on a fragment within the hypervariable region of 16S rDNA with the length of approximately 453 bp, the phylogenetic trees were constructed by Neighbor-Joining and Maximum-parsimony methods. The results indicated that the phylogenetic status of H. qinghaiensis was distant from that of H. longicornis and closer to H. flava. Furthermore, the ITS-2 rDNA was amplified by PCR and sequenced from individual ticks. The length of ITS-2 is 1,606 bp for H. longicornis and 1,162 bp for H. qinghaiensis. Although sequence variation between the immature stages of H. longicornis was 0.1–0.4%, nucleotide differences between the tested species ranged 2.1–23.2%, indicating that ITS-2 rDNA sequences are genetic markers for the differentiation of the two hard ticks in China. Hence, a PCR-linked restriction fragment length polymorphism (RFLP) approach was developed for their unequivocal differentiation based on ITS-2 rDNA, which provides the foundation for further studies on ticks in China and has implications for studying the population genetic structure of the ticks and for identification and differentiation of closely related ticks.     

Feeding patterns of immature stages of Hyalomma truncatum and Hyalomma marginatum rufipes on different hosts

In this study we examine the feeding patterns of immature stages of Hyalomma truncatum and Hyalomma marginatum rufipes ticks on different hosts. Larvae of H. truncatum developed through a three-host pattern on two species of field mice, Rhabdomys pumilio and Lemniscomys rosalia. On guinea-pigs, both Hyalomma species followed a mixed two-host and three-host pattern, with the latter route being preferred, since more than 70% of the fully fed larvae dropped off from their hosts. H. truncatum was a two-host tick on rabbits. Larvae of H. marginatum rufipes did not prefer R. pumilio and L. rosalia as hosts. On guinea-pigs, H. marginatum rufipes immatures showed a mixed two-host and three-host pattern with a bias towards the three-host life cycle, since approximately 58% of the fully fed larvae dropped off. On rabbits, H. marginatum rufipes was exclusively a two-host tick. Mean engorgement weights and blood quantities ingested by H. truncatum nymphs that developed through a three-host pattern on mice were significantly higher (p<0.0001) than for those that developed through a two-host pattern on guinea-pigs and rabbits. For H.marginatum rufipes, there were no significant differences (p>0.05) between engorgement weights of nymphs that developed through two-host and three-host patterns. However, there were significant differences (p<0.0001) in blood quantities ingested by nymphs of this tick species following feeding on different hosts.     

硬蜱中蜕皮激素含量的变化(英语)

为阐明蜕皮激素在硬蜱发育和生殖中的作用,用高效液相色谱法(HPLC)和放射免疫测定法测定了银盾革蜱和长角血蜱中饱血和产卵前后雌虫整体、血淋巴、合神经节及卵巢中蜕皮激素含量的变化.此外,还用HPLC测定了长角血蜱的卵和幼虫中蜕皮激素的含量.结果表明:在饱血前雌虫整体中蜕皮激素含量的变化不大,饱血后迅速增加.血淋巴中的蜕皮激素在饱血后第5天(即产卵前1天)达到高峰,可做为产卵的信号.合神经节中蜕皮激素含量的高峰在饱血后第5天,与血淋巴中高峰出现的时间一致,这与神经分泌细胞的分泌活性变化相吻合.从饱血后第3天起,卵巢中蜕皮激素含量增加很快,直到产卵时止,在长角血蜱的卵中也测到α-蜕皮素,来源于卵巢,随胚胎发育进程其含量逐渐增加.卵产出后第4天到第12天,蜕皮激素的含量增加十几倍.     

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.