A new species of Languidipes Hubbard (Ephemeroptera, Polymitarcyidae) from Borneo.

The genus Languidipes is currently represented by three species distributed in southeastern Asia, India, and Sri Lanka. Languidipes corporaali is the most widely distributed species, and both, male and female imagos, as well as nymphs, are known. In contrast, the other species, L. taprobanes and L. lithophagus, are only known from nymphs. Here, we describe a new species, Languidipes janae sp nov, based on male imagos collected from Borneo, Indonesia. This new species is characterized by the presence of ommation on mesonotum, and penis almost completely divided, with sub-quadrate base and a small outer projection basal to the long and slender distal arms. This constitutes the first record of the genus for Borneo. A cladistic analysis of the subfamily Asthenopodinae supports its taxonomic status.

Description of Rhipicephalus hibericus sp. nov. (Ixodoidea: Ixodidae), a species of the Rhipicephalus sanguineus group in southwestern Europe.

We describe all the life stages of Rhipicephalus hibericus n. sp., provide the types, and present molecular support for a new species of the Rhipicephalus sanguineus sensu lato group, present in southwestern Europe, that has been historically confused with Rhipicephalus turanicus Pomerantzev, 1940. A new name is proposed for this taxon because it was impossible to ascribe to types of already described species in the group, deposited for more than 100 years in natural history institutions. The males have a dorsum showing deep and coarse punctations (absent in Rhipicephalus sanguineus sensu stricto) and adanal plates with large punctations (absent in R. sanguineus s.s.); the tail of the spiracular plate is as wide as the closest festoon (half the width in R. sanguineus s.s.). Females have large punctations in dorsal fields, a wide spiracular plate, and a "V" shaped genital opening; such a combination of characters cannot be found in other species of the group. Immatures are described from specimens collected on hosts (Rodentia and Eulipotyphla). Both larvae and nymphs are markedly smaller than R. sanguineus s.s. Nymphs display long, backward-projected auriculae; larvae are almost half the size of R. sanguineus s.s. The new species can hybridize with R. sanguineus s.s. in laboratory colonies producing an unfertile F2, laying brown and dry eggs that did not hatch. Phylogenetic analysis of partial coxI gene sequences placed R. hibericus in a well-supported clade with other sequences of R. sanguineus s.l. from Portugal, as a sister clade of R. sanguineus s.s. The new species does not belong to the R. turanicus group of species. Both 12S and 16S partial gene sequences were not as precise in the correct phylogenetic placement of R. hibericus, in part probably due to the existence of erroneously identified sequences in GenBank©. This description, together with the previous reinstatement of Rhipicephalus secundus and Rhipicephalus rutilus, and the description of the neotypes of R. sanguineus s.s. should help researchers to adequately identify their collections. Our findings demonstrate that R. turanicus is absent in southwestern Europe. Old collections should be re-examined to provide the actual range of the new species.

Sensory pathway in aquatic basal polyneoptera: Antennal sensilla and brain morphology in stoneflies.

Aquatic insects represent a great portion of Arthropod diversity and the major fauna in inland waters. The sensory biology and neuroanatomy of these insects are, however, poorly investigated. This research aims to describe the antennal sensilla of nymphs of the stonefly Dinocras cephalotes using scanning electron microscopy and comparing them with the adult sensilla. Besides, central antennal pathways in nymphs and adults are investigated by neuron mass-tracing with tetramethylrhodamine, and their brain structures are visualized with an anti-synapsin antibody. No dramatic changes occur in the antennal sensilla during nymphal development, while antennal sensilla profoundly change from nymphs to adults when switching from an aquatic to an aerial lifestyle. However, similar brain structures are used in nymphs and adults to process diverging sensory information, perceived through different sensilla in water and air. These data provide valuable insights into the evolution of aquatic heterometabolous insects, maintaining a functional sensory system throughout development, including a distinct adaptation of the peripheral olfactory systems during the transition from detection of water-soluble chemicals to volatile compounds in the air. From a conservation biology perspective, the present data contribute to a better knowledge of the biology of stoneflies, which are very important bioindicators in rivers.

Revision of Thraulus Eaton 1881 (Ephemeroptera: Leptophlebiidae: Choroterpinae).

The genus Thraulus is widespread throughout much of the Eastern Hemisphere. Since Eaton established Thraulus in 1881, 62 species have, at one time or another, been placed in this genus. Thirty-eight of those species were eventually moved to other genera. Any comprehensive study of the remaining species, based on the published literature, is difficult as they were described by many authors, using different criteria, over a period of 142 years. The purpose of this study was to redescribe this genus, based on previously described species and nine new species, and to provide a format for future taxonomic and morphological studies of Thraulus. Redescriptions of most species were based on direct examination of external morphological characters. Descriptions or diagnoses of species, whose types were unavailable for study, were made using the original published description and additional information provided by authors of several of those species. The following species were studied: Thraulus amravati Vasanth, Subramanian & Selvakumar, 2022; T. bellus Eaton, 1881; T. bishopi Peters & Tsui, 1972; T. cuspidatus Vasanth, Subramanian & Selvakumar, 2022; T. demoulini Peters & Tsui, 1973; T. fasciatus (Kimmins, 1956); T. fatuus Kang & Yang, 1994; T. femoratus Li, Liu & Zhou, 2006; T. gopalani Grant & Sivaramakrishnan, 1985; T. jacobusi Isack, Srinivasan, Sivaruban & Barathy, 2022; T. macilentus Kang & Yang, 1994; T. malabarensis Vasanth, Subramanian & Selvakumar, 2022; T. mudumalaiensis Soman, 1991; T. plumeus Selvakumar, Vasanth & Subramanian, 2022; T. semicastaneus (Gillies, 1951); T. thiagarajani Balasubramanian & Muthukatturaja, 2019; T. thraker Jacob, 1988; T. torrentis (Gillies, 1964); T. turbinatus (Ulmer, 1909); T. umbrosus Kang & Yang, 1994; and T. vellimalaiensis Vasanth, Subramanian & Selvakumar, 2022. Nine new species of Thraulus are described: T. connubialis sp. nov., Malaysia; T. cursus sp. nov., Japan; T. eatoni sp. nov., Indonesia; T. ishiwatai sp. nov., Japan; T. madagasikarensis sp. nov., Madagascar; T. nihonensis sp. nov., Japan; T. opifer sp. nov., Australia; T. parentalis sp. nov., Malaysia; and T. petersorum sp. nov., Malaysia. Thraulus can be distinguished from all other genera of Leptophlebiidae by the following combination of characters: In the imagos, 1) upper portion of eyes oval-suboval, major axes diverge anteriorly; 2) vein MA fork of fore wings symmetrical; 3) vein MP fork of fore wings asymmetricala cross vein connects base of MP2 to MP1, MP fork closer to base of wing than Rs fork; 4) strongly oblique cross vein extends between veins R4+5 and MA1 just apical to fork of vein MA; 5) 2 cubital intercalary veins in fore wings; 6) costal projection on hind wings well-developed, bluntly rounded to acutely pointed; 7) claws dissimilarone blunt and pad-like, the other apically hooked; 8) penes long, relatively straight, narrow, parallel, usually contiguous mesally but not fused, apex may have lateral projections; 9) sternum 7 of female with posterior margin straight or shallowly concave or convex mesally; and 10) sternum 9 of females rounded apically. In addition, penile spines occur on most species. In the nymphs, 1) lateral margins of clypeus parallel; 2) width of labrum subequal to width of clypeus; 3) 2 dorsal rows of setae on labrum; 4) venter of labrum with 1 row of short stout setae on either side of midline near anterior margin, rows curve mesally; 5) hypopharynx with small, rounded, posterolateral projections on arms of superlingua; 6) large spine on posterolateral corners of terga 69, 79 or 89; 7) gills 17 dissimilar: gill 1 composed of 1 or 2 subulate lamellae or a dorsal subulate lamella and a ventral fimbriate oval lamella, and gills 27 composed of dorsal and ventral oval lamellae with fimbriate margins. Two species continue to be nomen dubiumT. siewertii (Weyenbergh, 1883) and T. vogleri (Weyenbergh, 1883). Thraulus grandis Gose, 1980 is considered nomen nudum. A review of published phylogenetic studies involving Thraulus is provided. With the species discussed in this paper, along with reports of additional new species to be described, Thraulus has the potential to be included among the more specious genera of Ephemeroptera.

Microscopy-based morphological characterization of rugose spiraling whitefly, (Aleurodicus rugioperculatus Martin)-an exotic pest on coconut in India.

Precise identification of exotic whiteflies is a prerequisite to curb the invasive potential on to a new geographical location and to evolve effective management strategies. Conventionally, whitefly taxonomy is based on the description of the fourth-instar nymph or puparium, however, in the current investigation egg to adult morphology along with morphometrics of rugose spiraling whitefly (RSW), Aleurodicus rugioperculatus Martin was critically analyzed and various morphological characteristics were illustrated. The morphometric analysis revealed that in the immature life stages of the pest, the length: width ratio decreased with each successive life stage, such as the egg, first, second, third, and fourth instar (2.472 ± 0.071, 1.913 ± 0.020, 1.550 ± 0.045, 1.297 ± 0.034, and 1.174 ± 0.058 mm), respectively. Across different nymphal instars, the shape of lingula was greatly modified from tongue-like to triangle-shaped. The number and distribution of compound pores also vary among the different nymphal stages. The ultra-structures of the antenna through electron-microscopy depicted finer details of sensory cones. The modulation in the shape and the structural arrangement of microtrichia on the plate was illustrated. The study indicated accurate diagnosis of various stages of RSW for effective interception of goods at quarantine stations and thus preventing the entry of exotic pests into the country. RESEARCH HIGHLIGHTS: Microscopy-based (light and scanning electron microscopy) morphological characterization of rugose spiraling whitefly, (Aleurodicus rugioperculatus Martin).

Morphology of the preadult stages of Coronaproctus castanopsis Li, Xu & Wu (Hemiptera: Coccomorpha: Monophlebidae).

All the preadult stages of Coronaproctus castanopsis Li, Xu & Wu, 2023 are described and illustrated (first- and second-instar nymphs, third-instar male and female nymphs, and the male pupa), based on material from Zhejiang Province, China. A key to separate the developmental stages is provided.

A new species of Megaglena Peters amp; Edmunds, 1970 (Ephemeroptera: Leptophlebiidae) from Tamil Nadu, India.

Megaglena sivarubani sp. nov. is described based on the nymphs from the Puliyuthu falls of Tamil Nadu, Southern India. The number of species of Megaglena Peters Edmunds is now expanded to three and it is the first record of the genus from the Tamil Nadu part of Southern Western Ghats. Megaglena sivarubani sp. nov. can be distinguished from Indian species M. agasthiya Vasanth, Subramanian Selvakumar by the body size of the nymphs, shape of the labial palp segment III, and by the setation of foretibia. The morphological variations between all the known species of Megaglena are discussed and summarized.

Morphological study of nymphal stages in Tenodera fasciata (Olivier, 1792) (Mantodea: Mantidae).

To date, no accurate identification method has been established for distinguishing mantis species of the genus Tenodera at all instar nymphal stages. In this paper, all instar nymphal stages of Tenodera fasciata (Olivier, 1792) are described in detail. This is the first step toward establishing an identification method for distinguishing the species from other members of the genus, particularly, T. angustipennis Saussure, 1869, which is sympatric with T. fasciata in Japan. Morphological differences between instars in nymphal stages, and between male and female in nymphal stages, were found. However, there was no difference between male and female 1st instars. Sex in nymphal stages are identified by the presence or absence of a crack at the posterior margin of the abdominal sternite IX between immature gonoplacs, the shape of the posterior margin of the sternites VII and IX, the occurrence of gonapophyses on the sternite VIII, and so on. Instars in nymphal stages are identified by body length, the length of discoidal spines of the profemur, the shape and size of the wing pads, the shape of abdominal sternite VII to IX, and so on.

Body shape and fluctuating asymmetry following different feeding sources and feeding time in a triatomine, Triatoma pallidipennis (Stål, 1892).

Even when an animal has a generalist diet, different food sources can impact its body shape and fluctuating asymmetry (a stress indicator; FA). To test this, we varied the food source (mammalian, avian or defibrinated mammalian blood; and control animals - ad libitum feeding) and the time of feeding (every 8 days, 45 days and ad libitum) having the Chagas triatomine vector, Triatoma pallidipennis (Stål, 1892), as a study animal which has presumable generalist feeding habits. This factorial design was applied since first instar animals until adulthood. As response variables, we measured body shape and FA in adults of both sexes, using a two-dimensional geometric morphometrics protocol. The highest variance in body shape was explained by diet (17%), followed by sex nested within diet (12%). Males had less morphological differentiation than females: females with defibrinated blood provided every 45 days differentiated more, while those that fed on mammalian blood every 8 days differed less. Distances among the averages of the FA component related to shape indicated greater distances between avian blood provided every 45 days and mammalian blood provided every 8 days, as well as between the two groups fed on avian blood (feeding every 8 and 45 days), followed by avian and defibrinated blood, both fed every 8 days. These results indicate that blood source and feeding time have significant effects on the body shape, and FA in females and both sexes. Thus, despite general feeding habits, avian blood showed a greater impact on shape and FA in triatomines. This may select for triatomines to use mammal blood rather than avian blood if they have the chance to do so.

Five ixodid tick species including two morphotypes of Rhipicephalus turanicus on nestlings of Eurasian eagle owl (Bubo bubo) from south-eastern Bulgaria.

BACKGROUND: Birds are major hosts for many tick species (Acari: Ixodidae, Argasidae), and their role is especially important in transporting ticks over large distances along their seasonal migratory routes. Accordingly, most studies across Europe focus on the importance of avian hosts in tick dispersal, and less emphasis is laid on resident birds and their role in supporting tick life cycles. Eurasian eagle owls (Bubo bubo) exemplify the latter, but all the few studies on their tick infestation were carried out in Western Europe and even those did not involve a large sample size and did not assess infestation prevalence in natural habitats. METHODS: In this study, 320 ixodid ticks were collected from nestlings of Eurasian eagle owls during the period 2018-2020 in Bulgaria in south-eastern Europe. These ticks were analysed morphologically, and selected specimens molecularly based on cytochrome c oxidase subunit I (cox1) gene. The effects of environmental and habitat-related conditions and of the species of prey eaten by eagle owls on tick infestation were also evaluated. RESULTS: The majority of ticks were identified as adults of Rhipicephalus turanicus (n = 296). In addition, 15 Hyalomma marginatum (three males, 11 nymphs and a larva), one female of Haemaphysalis erinacei and of Ha. punctata, and a nymph of Ixodes ricinus were found. Among R. turanicus, two distinct morphotypes were observed, but they do not form a monophyletic clade in the phylogenetic tree based on the mitochondrial gene cox1. We found a positive correlation between the total number of ticks on nestlings from a particular nest and the number of medium-sized to large prey mammals brought to the nestling owls. Also, the most important predictor for tick abundance was the effect of the extent of arable land (negative), while forests and grasslands contributed less, with no effect observed in case of urbanized areas and watercourses. CONCLUSIONS: The intensity of tick infestation can be high on nestling Eurasian eagle owls (mean intensity 16.59 ticks/nestling). In this study, five different tick species were recorded, among which R. turanicus dominated. Two male morphotypes of this tick species were found, but their morphological differences were not reflected by genetic diversity or phylogenetic clustering. The most important factor determining tick abundance was the land-use structure.

Double anus in an Ixodes scapularis nymph, a medically important tick vector.

BACKGROUND: Ixodes scapularis ticks are medically important arthropod vectors that transmit several pathogens to humans. The observations of morphological abnormalities, including nanism, missing leg, extra leg, and gynandromorphism, have been reported in these ticks. In this study, we report the presence of two anuses in a laboratory-reared I. scapularis nymph. RESULTS: Larval ticks were allowed to feed on mice and to molt to nymphs. Two anuses were observed in one of the freshly molted nymphs. Stereo and scanning electron microscopy confirmed the presence of two anuses in one nymph within a single anal groove. CONCLUSIONS: This report confirms the rare occurrence of double anus in I. scapularis.

Rediscovery and first genetic description of some poorly known tick species: Haemaphysalis kopetdaghica Kerbabaev, 1962 and Dermacentor raskemensis Pomerantzev, 1946.

This study aimed to provide novel information for some poorly known/rare tick species collected from wild goats (Capra aegagrus) in the mountains of Eastern Anatolia, Turkey and to expand upon the available genetic data. The collected ticks were morphologically identified as Haemaphysalis kopetdaghica (all active stages, n = 140), Dermacentor raskemensis (adults, n = 7), Ixodes gibbosus (adults, n = 15), Rhipicephalus kohlsi (female, n = 1), and R. bursa (nymphs, n = 2). A total of 32 engorged ticks (6 larvae, 6 nymphs, and 20 females) collected were allowed to molt to the next stage or for egg laying and larval hatching, respectively. In addition, one R. kohlsi female (previously confirmed by SEM microscopy) collected from a wild goat in the neighboring province of Erzurum was included in this study for further genetic comparison. The partial mitochondrial 16S rDNA and cytochrome c oxidase subunit 1 (barcoding regions) genes of each tick species were sequenced. All DNA samples obtained from the ticks were checked by PCR for the presence of Anaplasma spp., Babesia spp., Borrelia burgdorferi sensu lato, spotted fever group rickettsiae, and Theileria spp., but were found to be negative. Phylogenetic analyses of the 16S rDNA and cox1 genes were performed using the ML method to determine their genetic relationship with related ticks. As a result, this study has: i) rediscovered and provided two new tick records (H. kopetdaghica and D. raskemensis) for Turkey, ii) provided the first genetic data for H. kopetdaghica and D. raskemensis and revealed their phylogenetic relationships, iii) characterized the cox1 region of I. gibbosus for the first time, and iv) revealed significant genetic diversity between R. kohlsi from Anatolia and R. kohlsi from Oman, suggesting that R. kohlsi could include a cryptic species.

Morphology of Pholeoixodes species associated with carnivores in the western Palearctic: Pictorial key based on molecularly identified Ixodes (Ph.) canisuga, I. (Ph.) hexagonus and I. (Ph.) kaiseri males, nymphs and larvae.

Three Palearctic members of the subgenus Pholeoixodes, i.e., Ixodes canisuga, Ixodes hexagonus and Ixodes kaiseri are frequently collected from dogs, cats, red foxes, badgers and other carnivorous/insectivorous hosts in Europe. While a pictorial identification key has been reported for female Pholeoixodes ticks, a similar work has not been done on their male, nymphal and larval specimens. This study was initiated in order to clarify and re-examine those morphological characters of these three tick species, which can be used relatively easily to identify/distinguish them. In the case of larvae the aims included finding alternatives to chaetotaxy, which is difficult to observe and its usefulness is also affected by uncertainties in literature data. For this, 609 Pholeoixodes ticks (males, nymphs and larvae) were collected from carnivores, hedgehogs and their environment in six European countries (representing Western, Central and Southeastern Europe), followed by detailed morphological examination and/or molecular analyses to confirm the identity of their species. Based on the morphology of 84 molecularly analyzed specimens and a new identification key compiled accordingly, altogether 116 I. canisuga, 277 I. hexagonus and 216 I. kaiseri males, nymphs and larvae were identified. Ixodes kaiseri was not found in Western Europe, where I. canisuga predominated. In Central Europe, all three Pholeoixodes species were collected, the largest number of specimens represented by I. hexagonus. On the other hand, in Southeastern Europe I. kaiseri had the highest abundance. In conclusion, the morphology of internal spur on the first coxae (as the traditionally used character to distinguish I. hexagonus from other Pholeoixodes species) is trustworthy to recognize males but is less informative in the case of nymphs and larvae. The latter can be identified more properly by observing the morphology of basis capituli. In particular, nymphs and larvae of I. canisuga have anteriorly flattened basis capituli, forming a plateau that surrounds the base of the hypostome. On the other hand, nymphs and larvae of I. hexagonus and I. kaiseri lack a similar plateau, but (unlike I. canisuga) have cornuae, which are either posterolaterally or caudally directed, respectively.

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.

Ixodes silvanus n. sp. (Acari: Ixodidae), a new member of the subgenus Trichotoixodes Reznik, 1961 from northwestern Argentina.

Females, nymphs, and larvae of Ixodes silvanus n. sp. collected from birds and from the vegetation in northwestern Argentina (Yungas Phytogeographic Province) are described herein. The new species belongs to the subgenus Trichotoixodes (Acari: Ixodidae). The female is diagnosed by a combination of the following characters: scutum with setae moderately long and more numerous in central field, fewer and moderately long setae on lateral fields, and inconspicuous setae in anterior field; basis capituli subtriangular dorsally; porose areas large and irregular in shape, lacking distinct margins; auriculae with straight edges diverging posterolaterally and ending with small blunt processes; hypostome narrow and pointed with dental formula 4/4 in the anterior third, then 3/3 and 2/2 near the base; coxae I with two spurs, sub-equal in size, internal slightly slimmer than external. The nymph is diagnosed by notum with numerous and long setae, ventral surface covered by numerous whitish setae, scutum with short scapulae and few and shallow punctations, setae on scutum few, short and irregularly distributed, basis capituli sub-triangular dorsally with posterior margin straight, cornua large and directed postero-laterally, auriculae large and projected laterally, lateral margin of basis capituli above auriculae with a lateral and triangular projection, hypostome pointed with dental formula 3/3 in the anterior third and then 2/2, and coxa I with two short, sub-equal, triangular spurs. The diagnostic characters of the larva are: basis capituli dorsally sub-triangular with lateral angles acute and posterior margin straight, auriculae as large triangular lateral projections, hypostome with apex bluntly pointed and dental formula 3/3 in the anterior third and then 2/2, coxa I with two short, sub-equal, triangular spurs, and pattern of dorsal and ventral body setae. This new species is phylogenetically related to Ixodes brunneus, Ixodes turdus and Ixodes frontalis, and the principal hosts for all its parasitic stages are birds.

Morphological and Molecular Identification of Tropical Bed Bugs From Two Cities of the Pearl River Delta in China.

From the 1960s to the 1980s, with the implementation of nationwide 'Four-Pest Elimination' campaigns (bed bugs, Cimex lectularius L. and Cimex hemipterus (F.) (Hemiptera: Cimicidae), were listed as one of the targeted pests), bed bug infestations were gradually eliminated in most provinces in China. However, during the last two decades, reports of bed bug infestations in the Pearl River Delta of China have shown an upward trend. Up to now, the bed bug species occurring in this area was much less frequently reported. In this study, we used both morphological and molecular methods to accurately identify the species of bed bugs collected from the cities of Guangzhou and Foshan, China. Results indicated that no significant difference was observed in the mean pronotum width-to-length ratio of Guangzhou (2.6) and Foshan (2.4) specimens; however, both were significantly lower than that of a laboratory strain C. lectularius (3.1). The genetic distances of our specimens with C. hemipterus and C. lectularius were 0-0.2% and 22.2-22.6%, respectively. On the basis of the morphological characteristics and mitochondrial DNA sequence data, it can be affirmed that bed bugs collected from Guangzhou and Foshan were C. hemipterus.

Morphological and molecular characterization supporting Amblyomma mixtum presence in Cuba.

Amblyomma cajennense Fabricius, 1787 (Acari: Ixodidae) is a widely distributed tick taxon. Recent studies have reassessed this taxon as a complex of six species. Amblyomma mixtum Koch, 1844 has been suggested by some authors as the only species of this complex that is present in Cuba. Other authors have pointed a niche overlapping for A. mixtum and A. cajennense s.s. in the country. Detailed taxonomic studies on the Cuban species belonging to this complex are needed in order to evaluate their current distribution according to the recent classification. This study aimed to characterize Cuban populations from the A. cajennense complex by using tick samples obtained from 3 occidental provinces and 1 central province of the country. Morphological identification and measurements of the main relevant taxonomic structures were conducted by using Scanning Electron Microscopy. Phylogenetic analyzes were carried out with 16S ribosomal RNA, internal transcribed spacer 2 and the subunit I of mitochondrial cytochrome c oxidase gene sequences. The results of these studies demonstrated that all samples belonged to the species A. mixtum (Koch, 1844). This study constitutes the first molecular characterization of this Amblyomma species in Cuba. Further studies will be necessary in order to corroborate if A. cajennense s.s. is also present in the island.

Wax glands of the horned gall aphid, Schlechtendalia chinensis, at different stages.

The horned gall aphid, Schlechtendalia chinensis, inhabits the productive species of Chinese gallnuts, which have economic value. Aphid wax glands are crucial for the survival of the insects, since the secreted waterproofing wax is important to protect the aphids from predators, pathogens and honeydew contamination. In this study, we investigated the structure of wax glands and their role in different aphid stages using light and electron microscopy. Our results show that aphids of all stages except the newly hatched fundatrix possess six parallel dorsal lines and have a total of 56 wax gland plates. Although no wax glands occur on the dorsum of the newly hatched fundatrix (first instar), the glands do appear once a fundatrix enters the second instar. The wax gland plate is composed of 2-22 polygonal depressions, each of which corresponds to a secretory cell covered by cuticle. The wax glands of this aphid belong to the class 1 glands, which are formed by epidermal secretory cells. The structure of the wax glands varies in the different stages and these changes may be adaptive to the changeable microenvironments in which the aphids live.

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.

Exploring the prevalence and diversity of bovine ticks in five agro-ecological zones of Pakistan using phenetic and genetic tools.

Tick infestation is a leading cause of tick-worry and tick-borne diseases in livestock and associated economic losses in tropical and subtropical regions of the world. The cattle and buffalo populations in Pakistan are exposed to tick infestation throughout the year, but very little is known about the biology, diversity and distribution of tick species across different agro-ecological zones (AEZ) of the country. The present study aimed to investigate the prevalence (number of bovines infested with ticks out of the investigated population) and diversity of hard ticks infesting bovines in 30 villages located in five distinct AEZs (i.e. Arid, Indus delta, Northern irrigated plain, Sandy desert and Southern irrigated plain). We collected a total of 774 ticks (adult and nymphs) from cattle (n = 116) and water buffaloes (n = 88) on small-holder dairy farms (with <10 bovids per establishment) from September to November 2017. The overall tick prevalence was 46.1% (cattle: 47.9%; buffaloes: 44%), which varied significantly from 22.2% in the Indus delta to 70.5% in the Sandy desert. Tick prevalence was slightly higher in female (46.5%) than male animals (45%), and higher in calves (i.e. ≤ 1 year of age) (55%) than in young animals (i.e. up to 3 years of age) (39%) and adults (48%). Five tick species - Hyalomma anatolicum, Hyalomma hussaini, Hyalomma scupense, Rhipicephalus microplus and Rhipicephalus annulatus - were identified morphologically and then genetically. Genetic identification, achieved using the sequences of two mitochondrial (cytochrome c oxidase subunit 1 and 16S) and one nuclear ribosomal (second internal transcribed spacer) regions, was consistent with the morphological findings. Phylogenetic analyses of the DNA sequence data sets showed that the five species of tick identified here were closely related to the same species or closely related species from within and outside of Pakistan. Of five presently recognised taxa within the R. microplus complex, two were identified herein, including the R. microplus clade C and R. annulatus. This investigation provides the first genetic evidence of the occurrence of R. annulatus in Pakistan as well as Hy. hussaini and Hy. scupense in bovines specifically in the provinces of Sindh and Punjab, respectively. The present findings emphasise the importance of combining morphological and molecular approaches to study the diversity of ticks. Further longitudinal studies are required to establish seasonal variations in the prevalence and distribution of bovine ticks in different AEZs of Pakistan.