A molecular survey of Anaplasma , Ehrlichia , Bartonella and Theileria in ticks collected from southeastern China Abstract To investigate the prevalence of Anaplasma, Ehrlichia, Bartonella and Theileria, we collected ticks from small mammals in six counties of Zhejiang Province in southeastern China. Polymerase chain reaction (PCR) amplification was performed to test Anaplasma, Ehrlichia, Bartonella and Theileria in tick samples. Positive PCR products were sequenced and then compared with previously published sequences deposited in GenBank using BLAST. About 292 adult ticks were captured and the dominant tick species were Ixodes sinensis and Haemaphysalis longicornis. Overall, 34 ticks (11.6%) were tested positive for at least one pathogen of Anaplasma, Ehrlichia, Bartonella and Theileria. Rates of PCR-positivity to Anaplasma, Ehrlichia, Bartonella and Theileriawere 5.5, 1.7, 2.4 and 2.4%, respectively. Positive rates of Anaplasma, Bartonella and Theileria were significantly different among ticks of different species. Prevalence of Anaplasma and Theileria varied significantly among ticks of different counties. Anaplasma, Ehrlichia, Bartonella and Theileria were widely prevalent in ticks in Zhejiang Province suggesting other tick-borne pathogens should also be suspected if patients had history of tick bites.

Seasonal occurrence and development of three closely related Oligonychus species (Acari: Tetranychidae) and their associated natural enemies on fagaceous trees Abstract We compared the life cycles and diapause attributes among three closely related spider mites, Oligonychus castaneae on Castanea crenata, and O. gotohi and O. amiensis on Lithocarpus edulis. The lower thermal thresholds from egg to egg were 10.5, 8.5 and 8.9 °C, respectively, and the thermal constants were 177.8, 229.5 and 232.5 degree-days, respectively. The cumulative hatching rates of diapause eggs of O. castaneae and O. gotohi increased as the season progressed in and after early-to-mid January, which indicates diapause termination. In contrast, O. amiensis showed higher hatching rates in December and January, but hatchability gradually decreased in and after February because some of the eggs died from the cold. Oligonychus castaneae and O. gotohifemales produced diapause eggs in response to the short photoperiod in late September to early October and in early-to-late October, respectively, which corresponded to the times predicted by the critical photoperiods (at 15 °C) of 12 h 15 min and 11 h 15 min for the respective species. Oligonychus castaneae showed at least a single population peak over the 3-year observation period, but the time of peak population varied from mid-July to mid-September. The population of O. gotohi was higher between November and May when diapause eggs were present on host plants in early winter and the first-generation females laid eggs on leaves in spring. The population of O. amiensis, which is a non-diapause species, was only high between September and December, because eggs were laid on leaves in autumn to winter and then gradually disappeared and/or died during winter. Natural enemies were observed as the number of spider mites declined, and the density suppression effect by natural enemies was confirmed in the field.

Spatiotemporal associations between the mite Brevipalpus yothersi and citrus leprosis virus C in orange orchards Abstract Citrus leprosis virus C (CiLV-C) is an economically important pathogen and the main causative agent of leprosis disease in citrus orchards. The main vector of this disease, the mite Brevipalpus yothersi, is widely distributed in Mexican orchards on a wide range of citrus species. Despite the importance of both the virus and the mite, field studies recording their occurrence and co-occurrence are practically non-existent. We systematically sampled orange orchards for both CiLV-C and B. yothersi throughout the year. The distribution of the CiLV-C and B. yothersi was evaluated on each sampling occasion and their spatiotemporal associations were determined. Specifically, 100–112 orange trees, distributed in 18 rows (five or six trees per row), were sampled monthly between March 2017 and February 2018 (11 sampling dates). Twenty leaves per tree were sampled on each occasion. The number of mites per tree and the percentage of leaves per tree with disease symptoms were recorded. On each sampling occasion, spatiotemporal associations between mites and disease were determined using the Spatial Analysis by Distance Indices (SADIE) method. CiLV-C and B. yothersi were identified using molecular methods. Throughout the study, the distribution of CiLV-C was aggregated and the distribution of B. yothersi was random. No association was found between the virus and the mite on any of the sampling dates. In total, 173 mites were collected, but only 43 mites were found to be carrying CiLV-C. The reason for this lack of association between the virus and the mite, as well as the impact of our findings on the epidemiology of the disease in orange orchards, are discussed.

Production of winter eggs in Schizotetranychus brevisetosus (Acari: Tetranychidae) inhabiting evergreen Japanese blue oak Abstract The overwintering pattern of parasitic herbivorous arthropods is closely related to host phenology, because defoliation imposes strong selection pressures on various developmental stages. This relationship has been well studied in populations of spider mites (Acari: Tetranychidae) on deciduous hosts, but is little studied in populations on evergreen hosts, probably because their leaves are always available. However, spring defoliation may also influence the life cycle. We studied the overwintering pattern of Schizotetranychus brevisetosus (Acari: Tetranychidae), a specialist on evergreen oak, Quercus glauca, in Kochi, Japan. Only adult females and their eggs (winter eggs) survived the coldest months. We also observed a conspicuous seasonal change in egg colour and size: December winter eggs were 1.7× larger than September summer eggs, suggesting the winter eggs are diapausing. Adult females produced summer eggs until November and winter eggs from late November until they disappeared in March. The winter eggs hatched in early March. The immature stages developed in mid- to late March, when 39% of old leaves fell, some of which carried hundreds of immature individuals. Therefore, even in evergreen hosts, defoliation places potentially strong selection pressure on the immature stages. Despite this, S. brevisetosus appears not to change its egg hatching season to avoid the defoliation season, so it may have evolved a behavioural adaptation to escape falling leaves. The existence of several species with similar lifestyles suggests the importance of host phenology and predation pressure for evolution of the overwintering pattern.

Life table parameters of the red palm mite Raoiella indica (Acari: Tenuipalpidae) at various temperatures and for sexual and asexual reproduction Abstract The red palm mite, Raoiella indica Hirst, is a threat to coconut, banana and native Arecaceae and Heliconiaceae in Brazil. This mite originated in the Eastern Hemisphere and was first reported in 2004 in the Americas, where the pest is spreading quickly and causing severe damage to its host plants. The objective of this work was to determine the life-history parameters of R. indica at constant temperatures, estimate its thermal requirements [threshold temperature (Tb) and thermal constant (K)] and also compare its life table parameters between sexual reproduction and parthenogenesis. The life tables were constructed on leaflets of Adonidia merrillii at 15, 20, 24, 27, 30 and 34 °C and 65% RH and a 12-h photoperiod. The longevity and the number of laid eggs of non-copulated adult females were evaluated at 27 °C. Raoiella indica had complete development, from egg to adult, only at 20, 24, 27 and 30 °C. At 15 °C, the eggs did not hatch, and at 34 °C, the mites survived only until the larval stage. For sexual reproduction, the optimal temperature was 27 °C, under which the reproductive parameters were higher. The reproductive parameters for sexual reproduction were higher than those for parthenogenesis. The Tb was 14.79 °C, and the thermal constant was 208.33 degree days. The life parameters estimated in this study can be used for modelling and predicting the population growth of R. indica in the field and consequently for improving their management strategies.

Rhipicephalus microplus , babesiosis and anaplasmosis in Uruguay: current situation and control or elimination programs on farms Abstract Rhipicephalus microplus causes high economic losses for Uruguay; the country is divided into a R. microplus-free area and an endemic area. The aim of this research was to evaluate the R. microplus and tick fever situation in Uruguay and the feasibility of implementing R. microplus control or elimination programs to reduce the number of infested farms. A representative survey was performed to determine the number of infested farms, the number of farms with outbreaks of tick fever and the control and preventive measures being used. Control or elimination programs using generational treatment were implemented on 30 farms. Previously, a situation diagnosis was performed by evaluating the background of each farm, identifying acaricide resistance, serologically evaluating the enzootic stability of tick fever, and analyzing the risk of R. microplus introduction to farms. According to the survey, R. microplus was present on 9544 (36%) farms, and 3436 (36%) of these farms experienced outbreaks of tick fever. Only 323 (9.4%) farmers used the tick fever vaccine. Of the 30 farms for which control or elimination programs were carried out, 18 reduced the number of treatments applied per year and six achieved the elimination of the cattle tick. It is concluded that by administering elimination or control programs, it is possible to reduce the number of infested farms and the number of treatments to maintain control. New surveys should be performed to measure the degree of technology adoption by farmers and to analyze the advances in tick control or elimination programs on farms.

Inter- and intraspecific variability of morphological and molecular characters in Allothrombium species, with special reference to Allothrombium fuliginosum Abstract Morphology-based identification of Allothrombium spp., in view of the limited knowledge of intraspecific variation, hinders the recognition of species borders and affects the views on the actual distribution of species. Therefore, identification will benefit from reference to molecular methods. The separate species identity of specimens putatively representing Allothrombium fuliginosum and A. pulvinum, both reported as widely distributed in the Palaearctic region and considered as potential biological control agents, was checked using morphological and molecular analyses. The representatives of various Allothrombium spp. collected in the Palaearctic were included in the analysis in order to ascertain the distance between species. The results of the morphological examination, supported by statistical inference, along with the comparison of COI and/or ITS2 sequences, weaken the hypothesis of synoccurrence of both species in the Palaearctic region. Hence, we hypothesize that A. fuliginosum is widely distributed in the Palaearctic, whereas A. pulvinum should be regarded a Nearctic species.

RNAi mediated knockdown of RpL11 , RpS2 , and tra-2 led to reduced reproduction of Phytoseiulus persimilis Abstract Phytoseiulus persimilis is one of the most important biological control agents of spider mites. Multiple studies have been conducted on factors affecting its reproduction, but limited research on related molecular mechanisms has been carried out. In this study, RNA interference of three genes, ribosomal protein L11 (RpL11), ribosomal protein S2 (RpS2), and transformer-2 (tra-2), to newly emerged females were performed through oral delivery of double-stranded RNA, and knockdown of target genes was verified using qRT-PCR analysis. When RpL11 or RpS2 was interfered, 42 and 30% P. persimilis individuals either laid no egg or had no egg hatched, whereas the remaining females had their oviposition duration reduced by 31.8 and 49.9%, fecundity reduced by 48.1 and 67.8%, and egg hatching rate reduced by 20.4 and 22.4%, respectively. In addition, offspring sex ratios were significantly male biased especially at low fecundities. When tra-2 was interfered, no significant difference in fecundity was detected, but egg hatching rate reduced by 30.6%. This study verified the possibility of RNA interference in Phytoseiidae through oral delivery, and indicated that RpL11 and RpS2 are involved in egg formation, whereas tra-2 is involved in embryo development in P. persimilis. Phytoseiid mites have different sex determination pathways compared to insects. The present study provides data and evidence at molecular biological level for future research on reproduction and sex determination of phytoseiid mites.

Clinical gamasoidosis and antibody response in two patients infested with Ornithonyssus bursa (Acari: Gamasida: Macronyssidae) Abstract Blood-feeding ectoparasites constitute a growing burden for human and animal health, and animal production worldwide. In particular, mites (Acari: Gamasida) of the genera Dermanyssus (Dermanyssidae) and Ornithonyssus (Macronyssidae) infest birds and cause gamasoidosis in humans. The tropical fowl mite, Ornithonyssus bursa, is commonly found in tropical and subtropical countries but rarely reported in Europe. In this research we characterized the first two cases in Spain of clinical gamasoidosis diagnosed in patients infested with O. bursa, and investigated the IgE, IgM and IgG antibody response to mite proteins and the carbohydrate Galα1-3Galβ1-(3)4GlcNAc-R (α-Gal) involved in the tick-bite associated alpha-Gal syndrome (AGS). The results suggested that O. bursa is establishing across Mediterranean countries, and may increase the risk for gamasoidosis. The immune antibody response to mite proteins was higher for IgM and similar for IgE and IgG antibodies between patients and non-allergic control individuals exposed to mite or tick bites. The anti-α-Gal antibody levels were similar between patients and controls, a result supported by the absence of this carbohydrate in mites. These results suggested that mite bites do not correlate with antibody response to acarine proteins or α-Gal, and are not associated with the AGS.

Microhabitat distribution of arboreal oribatid mites (Oribatida), associated with the Siberian pine ( Pinus sibirica ) of Western Siberia Abstract The species composition of arboreal oribatid mites that live on Siberian pine trees (Pinus sibirica) in the forest-tundra of Western Siberia was examined, specifically of three Siberian pines from two distinct forest stands (six trees in total). Samples of litter were taken near the tree trunk, as well as samples of bark, branches and needles from the tree. In total 144 samples were taken, from which close to 5000 mites were extracted. From the arboreal samples, the mites were extract by heptane flotation. Three species of oribatid mites were recorded for the first time in Russia: Diapterobates brevidentatus, Eueremaeus trionus and Cultroribula berolina. The highest density and the highest dominance index of these species were recorded in arboreal microhabitats. Thirty-one species of oribatid mites were identified in total. No oribatid mites were recovered from the needles of Siberian pine. The density of oribatid mites did not significantly differ among various heights of the crown and trunk. The lowest density of mites was recorded on young branches without needles, whereas other branch hypothetical microhabitats did not significantly differ from each other in terms of oribatid density. The greatest Simpson diversity index was recorded in the plant litter near tree trunks. On trees, the diversity index decreased with the height of the trunk and with the distance of branch sections from the trunk. The dominant species and the degree of their dominance varied among microhabitats and forest stands. Additionally, a high level of dominance of a single oribatid species was observed on tree branches, as well as on the trunk bark located above the bole. In the two forest stands, these species were D. brevidentatus and Ameronothrus dubinini. Based on the analysis of oribatid communities, three microhabitats were identified in the first forest stand: the bole bark, the bark of the trunk above the bole, and the branches. In the second forest stand, bases of tree branches were identified as an additional microhabitat. Oribatids inhabit Siberian pine trees in the severe conditions of the forest-tundra. Arboreal oribatid communities of various microhabitats vary in their qualitative and quantitative characteristics. Also, arboreal and forest litter communities of oribatid mites vary significantly.