Morphological descriptions of the nymph and adults of Ornithodoros clarki, the larva and nymph of Ornithodoros rondoniensis, with notes on their phylogenetic relationships.

Based on tick specimens collected recently in Mexico, Nicaragua, Panama and Brazil, we provide morphological descriptions of the nymph and adults of Ornithodoros clarki Jones & Clifford, 1972 from the first three countries, and the larva and nymph of Ornithodoros rondoniensis (Labruna, Terassini, Camargo, Brandão, Ribeiro & Estrada-Peña, 2008) from Brazil. Also, an analysis of mitochondrial 16S rDNA sequences was performed to analyze the phylogenetic relationships of these tick species. Adults and nymphs of O. clarki and O. rondoniensis are unique among the Argasidae family by presenting exceptionally large spiracular plates with small goblets, and an integument with smooth polygonal mammillae. However, these two species are morphologically distinct based on specific patterns of coxal folds, idiosomal mammillae and pilosity, and female genital flap. In contrast, the larvae of O. clarki and O. rondoniensis are morphologically identical, except for a general larger size of the former species; this slight difference is corroborated by Principal Component Analysis (PCA) by using 40 morphometric variables. Phylogenetic analyses including 16S rDNA partial sequences of different Ornithodoros taxa from Central and South America indicate that O. rondoniensis from Brazil diverges from O. clarki from Mexico, Nicaragua and Panama. However, phylogenetic distance separating both alleged species is similar or slightly lower than the distances depicted for conspecific populations of a few other Ornithodoros species. Nonetheless, our primary criterion to maintain O. rondoniensis as a valid species is because its adult and nymphal stages do present distinct morphological traits that easily distinguish these postlarval stages from O. clarki.

Morphological and Developmental Traits of the Binucleation of Male Accessory Gland Cells in the Benthic Water Bug, Aphelocheirus vittatus (Hemiptera: Aphelochiridae).

The male accessory glands (MAGs) in insects are pair(s) of internal reproductive organs that produce and secrete the plasma component of seminal fluid. In various insects, MAG size is important for male reproductive success because the fluid provides physiologically active substances and/or nutrients to females to control sperm as well as female reproductive behaviors. Although the MAG epithelial cells in most insect species are standard mononucleate cells, those in some insect taxa are binucleate due to incomplete cytokinesis (e.g., Drosophila [Fallén] [Diptera: Drosophilidae]) or cell fusion (e.g., Cimex [Linnaeus] [Hemiptera: Cimicidae]). In the case of Drosophila, the apicobasal position of the two nuclei relative to the epithelial plane changes from vertical to horizontal after nutrient intake, which allows the volume of the MAG cavity to expand effectively. On the other hand, in the case of Cimex, the positions of the two nuclei do not change apicobasally in response to feeding, but their position relative to the proximodistal axis varies depending on the tubular/spherical organ morphology. Here, we report that the MAG of the benthic water bug Aphelocheirus vittatus (Matsumura) (Hemiptera: Aphelochiridae) shows binucleation in all epithelial cells. Despite the phylogenetically close relationship between Aphelocheirus and Cimex, the MAG cells in Aphelocheirus showed a Drosophila-like apicobasal change in the position of the two nuclei in response to feeding. Furthermore, the cytological processes during binucleation are more similar to those in Drosophila (incomplete cytokinesis) than to those in Cimex (cell fusion). These results indicate that the physiological role and mechanism of binucleation in MAG cells changed during the evolution of Hemiptera.

Redescription of the larva, and description of the nymphal and adult stages of Ornithodoros peruvianus Kohls, Clifford & Jones, 1969 (Acari: Argasidae).

The soft tick Ornithodoros peruvianus Kohls, Clifford & Jones, 1969 was described as a parasite of bats in Peru upon the examination of engorged larvae only. Recently, larvae of this tick species were reported on bats from northern Chile. However, the adult and nymphal stages of O. peruvianus have remained undescribed. This study aimed to redescribe the larva of O. peruvianus based on unfed specimens, and to describe nymphs, the male and the female of this species. Ticks were collected on the walls inside three caves in northern Chile. Two females laid eggs in the laboratory. Part of the unfed larvae was separated for morphological and morphometrical analyses, and the remaining specimens were fed upon laboratory mice in order to obtain subsequent nymphal and adult stages. The first nymphal stage (N1) moulted either to male or to a second nymphal stage (N2) without feeding. Obtained N2 moulted either to male or female after one meal. PCR amplification of tick mitochondrial 16S rRNA of specimens from the three caves revealed almost identical sequences. The unfed larva of O. peruvianus has an elongated idiosoma, and fringed setae cover the ventral surfaces of coxae, palps and tarsi. Nymph 1 has a thin integument covered by incipient mammillae and barely noticeable dorsal disks; it lacks cheeks and possesses few short setae on the basis capitulum. Nymph 2 has a pair of small cheeks and resembles adult stages in its tegumentary traits and capitulum. Adult stages exhibit developed cheeks (larger in females) without the capacity to completely cover the capitulum. Very small and low mammillae cover the surface of the dorsal idiosoma in adults. As this feature also occurs in other bat-associated soft ticks, regardless of their phylogenetic relatedness, small mammillae in bat soft ticks are suggestive of convergent evolution.

Evidence for a sexually selected function of the attachment system in bedbugs Cimex lectularius (Heteroptera, Cimicidae).

Attachment to surfaces is a major aspect of an animal's interaction with the environment. Consequently, shaping of the attachment system in relation to weight load and substrate is considered to have occurred mainly by natural selection. However, sexual selection may also be important because many animals attach to their partner during mating. The two hypotheses generate opposing predictions in species where males are smaller than females. Natural selection predicts that attachment ability will scale positively with load, and hence body size, and so will be larger in females than males. Sexual selection predicts attachment forces in males will be larger than those in females, despite the males' smaller size because males benefit from uninterrupted copulation by stronger attachment to the female. We tested these predictions in the common bedbug Cimex lectularius, a species in which both sexes, as well as nymphs, regularly carry large loads: blood meals of up to 3 times their body weight. By measuring attachment forces to smooth surfaces and analysing in situ fixed copulating pairs and the morphology of attachment devices, we show that: (i) males generate twice the attachment force of females, despite weighing 15% less; (ii) males adhere to females during copulation using hairy tibial adhesive pads; (iii) there are more setae, and more setae per unit area, in the pads of males than in those of females but there is no difference in the shape of the tarsal setae; and (iv) there is an absence of hairy tibial attachment pads and a low attachment force in nymphs. These results are consistent with a sexually selected function of attachment in bedbugs. Controlling sperm transfer and mate guarding by attaching to females during copulation may also shape the evolution of male attachment structures in other species. More generally, we hypothesise the existence of an arms race in terms of male attachment structures and female counterparts to impede attachment, which may result in a similar evolutionary diversification to male genitalia.

Instar Determination of Blattella asahinai (Blattodea: Ectobiidae) From Digital Measurements of the Pronotum Using Gaussian Mixture Modeling and the Number of Cercal Annuli.

The Asian cockroach, Blattella asahinai Mizukubo, has expanded its range throughout the southeastern United States since its introduction into Florida. Unlike its closest relative, the German cockroach, Blattella germanica (L.), B. asahinai lives outdoors and can fly. There is little information on the biology and development of B. asahinai, including the number of instars during nymphal development. To estimate the number of instars of B. asahinai, nymphs were photographed, sexed, and the lengths and widths of their pronota were measured digitally. The number of instars of B. asahinai was estimated using Gaussian mixture models with the pronotal data. The most probable model and its clusters were selected to assign individuals to an instar. Instars were also determined by counting the number of cercal annuli of nymphs. Both clustering and cercal annuli indicated that B. asahinai most frequently had six instars when reared at 30°C. Growth did not strictly follow the Brooks-Dyar Rule, because nymphs had different numbers of instars and different growth patterns. Although Gaussian mixture models are not efficient for field sampling experiments, digital measurements may provide a way to estimate instars with live specimens in development studies without handling the animals in a way that may alter growth.

Inter- and intraspecific variability of morphological and molecular characters in Allothrombium species, with special reference to Allothrombium fuliginosum.

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.

Data-driven predictions and novel hypotheses about zoonotic tick vectors from the genus Ixodes.

BACKGROUND: With the resurgence of tick-borne diseases such as Lyme disease and the emergence of new tick-borne pathogens such as Powassan virus, understanding what distinguishes vectors from non-vectors, and predicting undiscovered tick vectors is a crucial step towards mitigating disease risk in humans. We aimed to identify intrinsic traits that predict which Ixodes tick species are confirmed or strongly suspected to be vectors of zoonotic pathogens. METHODS: We focused on the well-studied tick genus Ixodes from which many species are known to transmit zoonotic diseases to humans. We apply generalized boosted regression to interrogate over 90 features for over 240 species of Ixodes ticks to learn what intrinsic features distinguish zoonotic vectors from non-vector species. In addition to better understanding the biological underpinnings of tick vectorial capacity, the model generates a per species probability of being a zoonotic vector on the basis of intrinsic biological similarity with known Ixodes vector species. RESULTS: Our model predicted vector status with over 91% accuracy, and identified 14 Ixodes species with high probabilities (80%) of transmitting infections from animal hosts to humans on the basis of their traits. Distinguishing characteristics of zoonotic tick vectors of Ixodes tick species include several anatomical structures that influence host seeking behavior and blood-feeding efficiency from a greater diversity of host species compared to non-vectors. CONCLUSIONS: Overall, these results suggest that zoonotic tick vectors are most likely to be those species where adult females hold a fecundity advantage by producing more eggs per clutch, which develop into larvae that feed on a greater diversity of host species compared to non-vector species. These larvae develop into nymphs whose anatomy are well suited for more efficient and longer feeding times on soft-bodied hosts compared to non-vectors, leading to larger adult females with greater fecundity. In addition to identifying novel, testable hypotheses about intrinsic features driving vectorial capacity across Ixodes tick species, our model identifies particular Ixodes species with the highest probability of carrying zoonotic diseases, offering specific targets for increased zoonotic investigation and surveillance.

Cryptic diversity within grass-associated Abacarus species complex (Acariformes: Eriophyidae), with the description of a new species, Abacarus plumiger n. sp.

Accurate estimation of species richness is often complex as genetic divergence is not always accompanied by appreciable morphological differentiation. In consequence, cryptic lineages or species evolve. Cryptic speciation is common especially in taxa characterized by small and simplified bodies, what makes their proper identification challenging. The cereal rust mite, Abacarus hystrix, was regarded for a long time as a species associated with a wide range of grass hosts, whereas wide host ranges are rather rare in eriophyoid mites. Therefore, the generalist status of A. hystrix was questioned. In this paper we demonstrate that the diversity within Abacarus species associated with grasses is more complex than it was previously thought. The 78 Abacarus mtDNA COI sequences used in this study formed 10 highly supported clades (bootstrap value 99%) and four more distinct genetic lineages were represented by unique sequences. The genetic distances between them ranged from 6.6 to 26.5%. Moreover, morphological study and genetic approach based on the combination of the Poisson Tree Processes model for species delimitation (PTP) and a Bayesian implementation of PTP (bPTP), and Neighbour Joining analyses led to delimitation of a new species within the Abacarus complex: Abacarus plumiger, specialized on smooth brome (Bromus inermis). Furthermore, our analyses demonstrated a pattern of host-associated differentiation within the complex. Overall, our study indicates that cryptic speciation occurs in the grass-associated Abacarus genus, and suggests the need for more extensive sampling using integrative methods.

Differential diagnosis of Ixodes ricinus and Ixodes persulcatus: nymphs and larvae.

We developed a method for differential diagnosis of nymphs and larvae of sheep (Ixodes ricinus (L.)) and taiga (I. persulcatus Sch.) ticks (Parasitiformes: Ixodidae) which allows to identify live material in the field.

Ixodes heathi n. sp. (Acari: Ixodidae), a co-endangered tick from the critically endangered mountain pygmy possum (Burramys parvus), with notes on its biology and conservation.

A new species of co-endangered tick, Ixodes heathi n. sp., is described from specimens of the nymph collected on the critically endangered mountain pygmy possum (Burramys parvus Broom) from the alpine region of Victoria, Australia. Its biology is discussed along with strategies for its conservation.

External morphology of the mouthparts and observations on behavior of Tuckerella japonica on Camellia sinensis in the continental USA.

Tuckerella japonica Ehara (Acari: Tetranychoidea: Tuckerellidae) was found on stems of seedling plants of Camellia sinensis (L.) O. Kuntze (Theaceae) in the continental USA. This mite is able to pierce exposed green periderm tissue with its paired stylets on 1- to 3- or 4-year-old stems where the outer bark had split longitudinally. The mite was not found on branches older than 3 or 4 years, where splitting diminished and, eventually, a uniform covering of bark was formed. The mouthparts of T. japonica were examined under scanning electron microscopy and their external morphology was compared with known Tetranychoidea. There were usually one or two feeding holes in an area where the female subsequently deposited one or more eggs. Females were observed defending the areas where their eggs and/or young occurred. This behavior along with potentially limited access to exposed periderm or availability of shoots in the rows of plants may restrict higher populations of T. japonica from becoming established. The potential of T. japonica as an economic pest remains unknown at this time.

A morphological and phylogenetic analysis of Ornithodoros marinkellei (Acari: Argasidae), with additional notes on habitat and host usage.

Ornithodoros marinkellei was described from larvae collected on Pteronotus spp. bats in Colombia and Panama. More recently, this tick was reported in the Brazilian Amazon. Because some morphometric differences were observed between O. marinkellei larvae from Colombia and Brazil, it was proposed that further investigations were needed to assess whether the differences could be attributed to intra- or inter-specific polymorphism. Herein, we collected O. marinkellei specimens in the type locality of Colombia, in Brazil, and in a new locality in Nicaragua, expanding the distribution of the species to Nicaragua. Morphometric analysis of larvae and adults, corroborated by a principal component analysis (PCA), indicated that the Brazilian specimens were larger than specimens from Colombia and Nicaragua. Phylogenetic analysis inferred from the mitochondrial 16S rRNA gene showed ticks from Colombia and Nicaragua more genetically related than any of them with ticks from Brazil, although ticks from the three countries grouped in a clade sister to a major clade containing sequences of various Neotropical Ornithodoros species. We concluded that ticks identified as O. marinkellei from Colombia, Nicaragua, and Brazil represent the same taxon, and that the genetic and morphological differences between them are likely to have a geographical bias. We redescribed the nymph of O. marinkellei, which has a vestigial hypostome, probably incompatible with blood feeding. We also report human infestation by O. marinkellei adults. As all reports of O. marinkellei adults have been from hot caves (temperature > 35 °C), this abiotic condition could be a limiting factor for the occurrence of this tick species.

New records of ticks infesting bats in Brazil, with observations on the first nymphal stage of Ornithodoros hasei.

In Brazil, at least 14 species of soft ticks (Argasidae) are associated with bats. While Ornithodoros hasei seems to be abundant among foliage-roosting bats, other groups of ticks are found exclusively inside caves. In this paper, noteworthy records of soft ticks infesting bats are documented in new localities from Bahia, Pernambuco, Piauí, and Rondônia states. Out of 201 bats examined, 25 were infested by 152 ticks belonging to seven taxa: Ornithodoros cavernicolous, O. hasei, Ornithodoros marinkellei, Ornithodoros cf. fonsecai, Ornithodoros cf. clarki, Antricola sp., and Nothoaspis amazoniensis. These findings provide new insights into the geographical distribution and host association of soft ticks occurring in the Neotropical region. Remarkably, morphological and biological observations about O. hasei are inferred based on the examination of on-host-collected first stage nymphs.

Redescription of the kiwi tick Ixodes anatis (Acari: Ixodidae) from New Zealand, with notes on its biology.

Ixodes anatis Chilton is of veterinary and conservation importance in New Zealand due to its association with the threatened kiwi (Aves: Apterygidae: Apteryx spp.). Since the tick's description in 1904, there has been debate regarding its correct taxonomic position and nomenclature. Herein all life stages of this species are redescribed and accompanied by electron micrographs and drawings. The systematics, host preferences, distribution, disease relationships and conservation biology of the species are also discussed.

Unexpected Infection with Armillifer Parasites.

Visceral pentastomiasis is usually found incidentally during surgery. We describe a case of visceral pentastomiasis discovered during inguinoscrotal hernia surgery for a man from Benin, Africa. Because surgical removal of nymphs is needed for symptomatic patients only, this patient's asymptomatic pentastomiasis was not treated and he recovered from surgery uneventfully.

Evaluating the submission of digital images as a method of surveillance for Ixodes scapularis ticks.

Widespread access to the internet is offering new possibilities for data collection in surveillance. We explore, in this study, the possibility of using an electronic tool to monitor occurrence of the tick vector of Lyme disease, Ixodes scapularis. The study aimed to compare the capacity for ticks to be identified in web-based submissions of digital images/photographs, to the traditional specimen-based identification method used by the provincial public health laboratory in Quebec, Canada. Forty-one veterinary clinics participated in the study by submitting digital images of ticks collected from pets via a website for image-based identification by an entomologist. The tick specimens were then sent to the provincial public health laboratory to be identified by the 'gold standard' method using a microscope. Of the images submitted online, 74·3% (284/382) were considered of high-enough quality to allow identification. The laboratory identified 382 tick specimens from seven different species, with I. scapularis representing 76% of the total submissions. Of the 284 ticks suitable for image-based species identification, 276 (97·2%) were correctly identified (Kappa statistic of 0·92, Z = 15·46, P < 0·001). This study demonstrates that image-based tick identification may be an accurate and useful method of detecting ticks for surveillance when images are of suitable quality.

Morphological anomalies in Ixodes ricinus and Ixodes inopinatus collected from tick-borne encephalitis natural foci in Central Europe.

In 2015, 0.02% nymphs displayed anomalies (ectromely) out of the 6744 collected ticks as part of a tick-borne encephalitis program in Germany. In 2016, questing Ixodes ricinus (n = 14,527) and Ixodes inopinatus (n = 75) ticks were collected by flagging as part of a tick-borne encephalitis program in Germany, Slovakia and Denmark. A total of 278 (1.9%) out of 14,602 nymph and adult ticks showed morphological anomalies. The anomalies were divided into general anomalies (body asymmetry) and local anomalies (anomalies of appendages, malformation of capitulum, exoskeleton anomalies and anal groove deformation) with nymphs being the most affected life stage. Most important, leg atrophy was the most common anomaly (209 nymphs, 11 females and three males) followed by asymmetry (10 nymphs, five females and a male) and ectromely (nine nymphs). Two females and one male displayed multiple anomalies on legs, palps and exoskeleton. Anal groove deformation was observed in three females and three nymphs. In 2016, the frequency of anomalies in I. inopinatus was found five times higher (9.3%) than in I. ricinus (1.9%). This is the first report of anomaly (ectromely, leg atrophy, idiosoma deformation) in flagged I. inopinatus and the first report of schizomely in I. ricinus.

Morphological and molecular identification of Rhipicephalus (Boophilus) microplus in Nigeria, West Africa: a threat to livestock health.

The cattle tick Rhipicephalus (Boophilus) microplus was first reported in West Africa in Ivory Coast, in 2007. Since then it has made an aggressive eastward advancement having been reported in four other West African countries: Mali, Burkina Faso, Togo and Benin. We herein report the first molecular identification of this tick species in Nigeria, West Africa, and highlight the threat it poses to livestock health. A nation-wide tick survey was conducted in 12 out of 36 states across the various agro ecological zones of Nigeria over a 1 year period (April 2014-March 2015). In total 1498 ticks belonging to three genera collected from cattle were morphologically identified. Overall, Amblyomma species constituted the highest percentage of sampled ticks, 50.2% (752/1498), followed by Rhipicephalus (including the subgenus Boophilus) species, 29.4% (440/1498) and Hyalomma species, 20.4% (306/1498). The presence of Rh. (B.) microplus was identified morphologically from four out of the 12 states. This finding was confirmed for the first time in Nigeria using a molecular method targeting the ITS-2 region of the ticks in three of the 12 states. This study ascertained the presence of Rh. (B.) microplus in Nigeria in addition to a broad variety of cattle tick species, most of which are of veterinary importance. The implication of this finding is that there may be additional economic burden to livestock farmers due to increased cost of tick control occasioned by the acaricide resistance by this tick species widely reported from different climes. Additionally, there may be a potential upsurge in incidence of hemoparasitic infections in cattle leading to increased morbidity, cost of treatment and mortalities.

[Immature stages of Amnestus rugosus (Hemiptera: Cydnidae)]. / Estadios ninfales de Amnestus rugosus (Hemiptera: Cydnidae).

In Mexico there are 29 species of Amnestus, of which six species are from Veracruz. The morphological features and illustrations of immature stages of A. rugosus are presented. Notes about the biology and behavior were included. The nymphal and adult stages of A. rugosus were found in remnants of riparian vegetation of the mesophilous forest in La Antigua river basin, Veracruz, Mexico. Twelve sites in five localities were selected and the specimens were sampled in rainy (July) and dry (April) seasons of 2012. The samples were processed in Berlese-Tullgren funnel and Winkler. A total of 419 individuals, 320 adults and 99 nymphs of A. rugosus were revised. The size and coloration of the body and punctuation patterns the immature stages of A. rugosus, A. ficus, and A. morelensis are the main differences among these species. A. rugosus is a first record from Veracruz.

Teratological Nymphal Ixodes scapularis (Acari: Ixodidae) From Wisconsin.

Abnormalities of physiological development (teratological forms) in ticks are rare. The occurrence of gigantism, dwarfism, gynandromorphs, missing legs, extra legs, and asymmetries is most often reported from lab-reared specimens, but has been observed in field-collected specimens. All morphologically anomalous ticks (besides gynandromorphy) described to date are from species other than Ixodes scapularis Say (Acari: Ixodidae). Here we describe four teratological I. scapularis nymphs collected while dragging vegetation in Wisconsin in 2015, including two asymmetrical ticks, one with a missing leg, and one with an extra leg.