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.     

Studies on in vitro Extrusion and Ultrastructure of the Spermatophore in Haemaphysalis longicornis (Acari: Ixodidae)

Copulation in ticks is completed by the insertion of the spermatophore into the female genital aperture by a male. The endospermatophore, a cord-like structure and contents are packed in the ectospermatophore of the completed spermatophore. The endospermatophore extrudes just after insertion of the tip of the spermatophore. Only the endospermatophore enters the female genital tract, and the ectospermatophore remains outside the female body. The extrusion is observed in vitro in Haemaphysalis longicornis at various concentrations of NaCl solution: the process is accelerated in less concentrated solutions. The cord-like structure and the endospermatophore finally receive contents extruded from the ectospermatophore. The tip of the cord-like structure connects to the surface of the endospermatophore, and together form a loop after extrusion. Ultrastructural observations confirmed that the ectospermatophore wall is composed of four layers, and the contents consist of male germ cells and three types of secretions from the male accessory genital glands. As in other ticks the male germ cells are elongated spermatids in spermatophores just after formation and extrusion. Adlerocysts described in other ticks are not found in the spermatophore of H. longicornis.     

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 identity of the type-species of Gonatocerus Nees (Hymenoptera: Mymaridae)

The validity of Gonatocerus Nees is discussed, and the identity of the type-species, G.longicornis, is established. Generic and specific synonymy is listed, and the neotype female of G.longicornis is redescribed.     

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.

     

The temperature and humidity preferences of Haemaphysalis longicornis,ixodes holocyclus and Rhipicephalus sanguineus (ixodidae): Studies on engorged larvae

The engorged larvae of Haemaphysalis longicornis, Ixodes holocyclus and Rhipicephalus sanguineus were exposed to a range of temperatures and humidities to see whether the nature of their requirements in the laboratory were similar to the climate within the geographic range of each species. The response of H. longicornis and I. holocyclus to changes in humidity was also studied. Moulting of I. holocyclus larvae occurred from 18 to 28°C and at a saturation deficit of 4 mm Hg or less. The larvae of R. sanguineus moulted between 18 and 38°C and tolerated saturation deficits up to 35 mm Hg. The larvae of H. longicornis moulted between 15 and 38°C at saturation deficits up to 8 mm Hg. When engorged larvae of H. longicornis and I. holocyclus were exposed to very dry conditions for different periods of time and then transferred to moist conditions, the minimum pre-moult period and mortality was increased in comparison with larvae kept continuously under moist conditions. On the other hand, mortality of H. longicornis larvae was reduced in very dry conditions provided that they had been exposed to a moist environment for about 5 days previously. The larvae of I. holocyclus required at least 12 days exposure to a moist environment before any survived to moult in moderately dry conditions. The engorged larvae of both H. longicornis and I. holocyclus lost weight rapidly in dry air, whereas weight loss from R. sanguineus larvae was much slower. The rate of development was fastest in R. sanguineus and slowest in I. holocyclus.     

Life cycle of <Emphasis Type="Italic">Haemaphysalis doenitzi</Emphasis> (Acari: Ixodidae) under laboratory conditions and its phylogeny based on mitochondrial 16S rDNA

The paper investigated the life cycle of the hard tick Haemaphysalis doenitzi under laboratory conditions and its phylogeny based on mitochondrial 16S rDNA. The results revealed that the complete life cycle of H. doenitzi requires a mean duration of 109.6 days ranging from 91 to 137 days and the average prefeeding, feeding and premoulting periods of larvae, nymphs and females and the eggs hatching period are 18.7, 26.9, 38.9, and 25.1 days, respectively. In addition, the weight of engorged females is highly correlated with the number of egg masses laid (r = 0.936, P < 0.001). The female reproductive efficiency index and reproductive fitness index are 13.4 and 12.8, respectively. The mean weight of the engorged nymphs (2.77 mg) moulting to females is much higher than those (1.68 mg) moulting to males, which could be used as an index to predict sexes in this species. The ratio of male to female is 1:1.01. Moreover, multisequence alignments and phylogenetic tree constructed based on the mitochondrial 16S rDNA sequences suggest that H. doenitzi is genetically close to H. longicornis.     

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.     

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.     

Bosmina (Eubosmina) longicornis longicornis SCHOEDLER, 1866 (Crustacea,Cladocera: Bosminidae) from the Postglacial Sediments of the Neuendorfer See,East-Germany

Analyses were made of subfossil Eubosmina remains from the Lake Neuendorfer See, a lake in the middle part of the river Spree in the “Urstromtal” of Baruth situated in the eastern part of Germany. In addition to Bosmina (Bosmina) longirostris (O. F. M., 1785) Bosmina (Eub.) longicornis longicornis Schoedler, 1866 occured in the lake from the Boreal (sediment depth 15.85 m) to the recent surface layers of the core. Other eubosminids did not occur during the history of the Lake Neuendorfer See with the exception of few remains of B. (Eub.) longispina, which were detected in the lowermost layers of the core during the late Pleistocene. In the present-day plankton, Bosmina (Eub.) l. longicornis was the only eubosminid found in the Lake Neuendorfer See. During the long period between colonization during the Boreal to the present, no changes were detected in typical morphological features of B. (Eub.) l. longicornis. All remains of Eubosmina in the layers above 15.85 m belong undoubtedly to the same taxon. No signs of any morphological instability (other then ontogenetic and cyclomorphic formchanges) or “morphological transition” (Hofmann, 1977) to other taxa could be found. By using paleolimnological analyses, the named taxon has proven to be a “good” entity within the group of long-spined taxa of the subgenus Eubosmina.     

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.     

Morphological characteristics and developmental changes of the ovary in the tick Haemaphysalis longicornis Neumann

Haemaphysalis longicornis (Ixodida: Ixodidae) is an important vector of transovarially transmitted parasites of the genus Babesia (Piroplasmida: Babesiidae). In the present study, we investigated the morphological characteristics and developmental changes of the ovary of H. longicornis. We show that the ovary of H. longicornis has a single tubular structure and is surrounded by a tunica propria. There is a longitudinal groove along one side of the ovary. During feeding and after engorgement, great changes can be observed in the ovary of H. longicornis and two rapid growth phases can be detected. The number of major protein bands of the ovary is significantly increased from day 3 of feeding and reaches a maximum on the day of engorgement. Therefore, the great diversity of proteins in the ovaries of H. longicornis can facilitate the identification of new targets for vaccine development.     

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.     

Reference gene selection for normalizing gene expression using quantitative real-time PCR in Haemaphysalis longicornis

The Asian longhorned tick, Haemaphysalis longicornis, the dominant species of Ixodidae in Korea, has a wide distribution in East Asia, far-East Russia, and Western Pacific countries, and has recently been discovered in the Eastern states of the United States of America. H. longicornis transmits various pathogens, including Babesia ovate, Rickettsia japonica, and severe fever with thrombocytopenia syndrome virus (SFTSV). Considering its medical importance, in order to understand the physiology of H. longicornis, it is crucial to determine the expression of the genes of interest. Although quantitative real-time PCR (qRT-PCR) has been widely used to analyze gene expression, stably-expressed internal reference genes across samples of different conditions should be selected for the accurate normalization of target gene expression levels. Therefore, in this study, we investigated the expression levels of five candidate reference genes, namely ACT, RPP0, RPL23, TUB, and GAPDH, in H. longicornis under different conditions, including different collection months, developmental stages, and SFTSV infection status. Using four software programs, namely, NormFinder, BestKeeper, geNorm, and RefFinder, their expression stabilities were evaluated. Subsequently, a single gene between RPL23 and RPP0 was validated, which was found to be most stable reference gene after comparing the expression levels of HSP70 determined using different normalization methods.     

Experimental evidence of habitat selection and territoriality in the Amazonian whip spider Heterophrynus longicornis (Arachnida, Amblypygi)

Animals that select and defend suitable habitats against conspecifics may be favored by maximizing prey encounter rate, gaining protection, or securing matings. However, the identification of habitat selection and territoriality may be hindered in observational studies in sedentary species with low-density populations, such as the whip spider Heterophrynus longicornis. To circumvent such difficulties, we adopted an experimental field approach to evaluate if H. longicornis selects and defends habitat in Central Amazon. To evaluate whether individuals perform habitat selection, we monitored the permanence of 29 experimentally released individuals in buttressed trees with a wide variation in diameter at breast height (DBH), with and without burrows at their bases. To evaluate whether individuals are territorial, we experimentally removed 21 individuals from their previously occupied trees and monitored the recolonization of these trees. If H. longicornis were territorial, we predicted that the recolonizers would be smaller than the removed individuals. We found 12 individuals in the trees where they were released on subsequent days, none of them in trees without burrows. The individual permanence was related to the presence of burrows, and not to DBH. There was recolonization by smaller males and females on ten trees from which the amblypygids were removed. In combination, the permanence of individuals in trees with burrows, the rapid recolonization of experimentally vacated trees by smaller individuals, and the preponderance of one individual per tree, suggest that both male and female H. longicornis perform habitat selection and also that after selecting a site, they defend it against conspecifics.     

Population studies of arthropods on Melia azedarach in Seville (Spain), with special reference to Eutetranychus orientalis (Acari: Tetranychidae) and its natural enemies

Eutetranychus orientalis has become an important pest of the ornamental tree Melia azedarach in the city of Seville (Spain). Trees suffer total defoliation at the end of summer. Studies were conducted in a regular plantation of this tree in the Miraflores Park in 2008 and 2009, to determine the arthropod faunal composition, with particular interest in the possible natural enemies of E. orientalis. Eutetranychus orientalis accounted for 98.3% of the arthropods found on the leaflets. Two species of phytoseiids were found, Euseius scutalis and Euseius stipulatus, but they only represented 0.2% of the arthropods. The most abundant insect was the predator thrips Scolothrips longicornis, which accounted for 0.9% of the arthropods found. The population of E. orientalis reached two peaks in 2008, with 325 individuals per leaflet in August, and 100 individuals per leaflet in November. Scolothrips longicornis densities closely followed E. orientalis, and predation was observed on various mite instars. Phytoseiids did not show such a response to the E. orientalis densities. Eutetranychus orientalis was more abundant in the exterior part of the plantation. No differences of arthropod densities were found between the various orientations in the plantation (north vs. south, east vs. west), although E. orientalis densities were different between rows. Distribution of E. orientalis population was highly aggregative, that of S. longicornis population was less aggregative, whereas the phytoseiid population showed a random distribution.     

Copepods grazing on <Emphasis Type="Italic">Coscinodiscus wailesii</Emphasis>: a question of size?

Grazing of female copepods, Acartia clausi and Temora longicornis, on the large centric diatom Coscinodiscus wailesii, was studied in the laboratory. While A. clausi females did not feed on C. wailesii, T. longicornis showed a skilful handling of these large algae. The cells were not completely ingested but copepods bit a small piece out of the silica wall and then ingested the cell content only. Faecal pellet analyses showed little remains of the silica walls, and most of the content could only be classified as undefined matter. Therefore the general analyses of faecal pellets and gut content of copepods from the field must be interpreted with caution.     

A revision of Hieracium (Asteraceae) in Norrbotten,northeastern‐most Sweden,revealed seven new species

The large quantity of material of Hieracium sectt. Hieracium, Vulgata, Bifida and Oreadea collected during the recently completed floristic inventory of the northeastern most Swedish province of Norrbotten has been revised. The number of species known from this previously badly under‐collected province hereby increased from 27 to 64, and seven species new to science were revealed. A diagnostic key to all 64 species is provided and H. alces T. Tyler, H. hystrix T. Tyler, H. larsii‐levii T. Tyler, H. pseudopsepharum T. Tyler, H. stenbergii T. Tyler, H. taedum T. Tyler and H. vexillatum T. Tyler are described anew.     

Trophic interactions between zooplankton andPhaeocystis cf.globosa

Mesozooplankton grazing onPhaeocystis cf.globosa was investigated by laboratory and field studies. Tests on 18 different species by means of laboratory incubation experiments, carried out at the Biologische Anstalt Helgoland, revealed thatPhaeocystis was ingested by 5 meroplanktonic and 6 holoplanktonic species; filtering and ingestion rates of the latter were determined. Among copepods, the highest feeding rates were found forCalanus helgolandicus andTemora longicornis. Copepods fed on all size-classes ofPhaeocystis offered (generally 4–500 μm equivalent spherical diameter [ESD]), but they preferred the colonies. FemaleC. helgolandicus and femaleT. longicornis preferably fed on larger colonies (ESD>200 μm and ESD>100 μm, respectively. However, a field study, carried out in the Marsdiep (Dutch Wadden Sea) showed phytoplankton grazing by the dominant copepodTemora longicornis to be negligible during thePhaeocystis spring bloom.T. longicornis gut fluorescence was inversely related toPhaeocystis dominance. The hypothesis has been put forward thatT. longicornis preferentially feeds on microzooplankton and by this may enhance rather than depressPhaeocystis blooms. Results from laboratory incubation experiments, including three trophic levels —Phaeocystis cf.globosa (algae),Strombidinopsis sp. (ciliate) andTemora longicornis (copepod) — support this hypothesis.     

Temperature-dependent functional response of Scolothrips longicornis (Thysanoptera: Thripidae) preying on Tetranychus urticae

Environmental variables like temperature are important factors that affect the efficiency of biological control agents in greenhouse crops. This study examined the effect of temperature on the functional response of an acarophagous thrips Scolothrips longicornis Priesner to different densities of two spotted spider mite, Tetranychus urticae Koch in laboratory conditions. Five constant temperatures (15, 20, 26, 30, and 35 °C) and seven prey densities (2, 4, 8, 16, 32, 64, and 128) were used in the experiments. At each temperature, 2 to 128 adult of T. urticaue were exposed to a female S. longicornis for a 24 h period. The results indicated that the predatory thrips exhibited type II functional response to different densities of T. urticae at all examined temperatures. Temperature had significant effect on the predation capacity of the adult thrips over the range of 15–35 °C. Instantaneous attack rate (a) of the predator increased linearly with increasing temperature from 15 to 35 °C. At 35 °C, the highest instantaneous attack rate was estimated to be 0.0579 day^? 1 using the Holling model and 0.1480 day^? 1 using Rogers model. The estimated value of handling time (T_h) using both Holling and Rogers models decreased linearly with increasing temperature from 15 to 35 °C. Scolothrips longicornis achieved higher predation at higher temperature (16.1 preys/day at 35 °C). This observation suggested that S. longicornis may be more effective for biological control of two spotted spider mite in warmer conditions.