The ontogeny of the cypridid ostracod Eucypris virens (Jurine, 1820) (Crustacea,Ostracoda)

The chaetotaxy (shape, structure and distribution of setae) of appendages and valve allometry during the post embryonic ontogeny of the cyprididine ostracod Eucypris virens are described. It is shown that the basic ontogenetic development of E. virens is very similar to that of other species of the family Cyprididae. During ontogeny, the chaetotaxy shows continual development on all podomeres of the limbs with the exception of the last podomere on the antennulae. The long setae on the exopodite and protopodite of the antennae have a natatory function until the actual natatory setae develop in later instars. Aesthetascs (presumed chemoreceptors) ya and y3 are the first to develop and may have an important function in the first instars. Cyprididae require a pediform limb in the posterior of the body presumably to help them to attach to substrates and this is reflected by the pediform nature of one limb at all times throughout all instars. This study has also shown that the fifth limb is most probably of thoracic origin and hence ostracods have only one pair of maxillae.     

Description of the Larval Stages of the Neotropical Water Beetle Hydraena particeps Perkins from Nicaragua

The three larval instars of Hydraena particeps Perkins, 1981 are described, based on material collected in the field and reared in the laboratory. Illustrations of structural features and chaetotaxy are provided. For the first time the three larval instars of one species of the genus Hydraena are described. Several characters are compared to those of Hydraena hernandoi Fresneda and Lagar from Europe and the North American Hydraena circulata Perkins.     

Immature stages of the genus Ectemnostega Enderlein (Hemiptera: Heteroptera: Corixidae), with an identification key to instars and redescription of the nymphs of E. (Ectemnostega) quadrata (Signoret)

The egg and the five instars of Ectemnostega are described based on examined material and published information. The characters most useful in identifying nymphs of Ectemnostega are body and head lengths and widths; number of transverse sulcations of the rostrum; chaetotaxy of mesofemur, meso- and metatibiae, metatarsus, and pterothorax; and grade of development of wing pads. A key to the five nymphal instars of Ectemnostega is provided. The egg of E. quadrata is figured and described for the first time; and the five instars are redescribed in detail and illustrated, with emphasis on the morphometry and chaetotaxy of selected structures.

Se describe el huevo y los cinco estadios de Ectemnostega a partir de material examinado e información publicada. Los caracteres mas útiles para identificar las ninfas de Ectemnostega son: longitudes y anchos de cuerpo y cabeza; número de: crestas transversales del rostro, quetotaxia del mesofemur, meso- y metatibias, metatarso, y pterotórax; y grado de desarrollo de pterotecas. Se provee una clave para identificar los cinco estadios de Ectemnostega. Por primera vez se ilustra y describe el huevo de E. quadrata; y se redescriben en detalle e ilustran los cinco estadios, con énfasis en la morfometría y quetotaxia de ciertas estructuras.     

Observations on cercarial chaetotaxy as a means for the identification of European species of Trichobilharzia Skrjabin & Zakharow, 1920 (Digenea: Schistosomatidae)

In order to evaluate the importance of cercarial chaetotaxy in the identification of European avian schistosomatids of the genus Trichobilharzia Skrjabin & Zakharow, 1920, papillary patterns of T. franki Müller & Kimmig, 1994 and T. ocellata (La Valette, 1855) Brumpt, 1931 were examined and compared. Scanning electron microscopy (SEM) was also employed to confirm the number and distribution of the sensory papillae. The chaetotaxy of T. franki and T. ocellata were identical, which suggests a close kinship between the two species. Some sensory papillae stained insufficiently with silver nitrate or were difficult to examine using light microscopy. Thus, the use of SEM in future examinations of Trichobilharzia chaetotaxy will ensure the collection of data that are comparable between studies, even though some argentophilic structures are not visible using SEM.     

The external morphology of the pseudoscorpion Roncus andreinii

The external morphology of the four postembryonic instars of Roncus andreinii (di Caporiacco, 1925) is described and compared with one of the related R. lubricus L. Koch, 1873 (Gabbutt & Vachon, 1967), with particular emphasis on disposition of the chelal trichobothria in relation to growth.
From the evaluation of characters it is concluded that a few morphometric and chaetotaxic characters (i.e. the length, ratio and shape of pedipalpal articles, the ratio of fingers/pedipalpal hand, the chaetotaxy of sternites II and III) represent good specific discriminants, while the relative position of trichobothria along the chelal axis is nearly the same in both species.
The localization of the growing and non-growing regions along the pedipalpal fingers may be of practical use in the search for new taxonomic characters.     

DNA identification and morphological description of the first confirmed larvae of Hetaeriinae (Coleoptera: Histeridae)

Abstract.  Using DNA sequences of nuclear ribosomal (18S) and mitochondrial (cytochrome oxidase I) genes (a modified DNA barcoding approach), we positively identify, for the first time, larvae of hetaeriine Histeridae. Species in this subfamily occur as obligate associates of social insect colonies, particularly those of neotropical army ants. Of several larval specimens collected from bivouacs of Eciton burchelli , we identify the two larval instars of Paratropinus scalptus , and discuss a quite different first instar larva near Euxenister . The larvae are described and illustrated, with attempts to homologize all chaetotaxy to other known histerid larvae. Phylogenetic trees for 18S and cytochrome oxidase I, for over twenty hetaeriine taxa, are compared with each other and with a previous hypothesis of relationships in the subfamily.     

Natural History and Comparative Morphology of Immatures of <Emphasis Type="Italic">Gamelia anableps</Emphasis> (C. Felder &amp; R. Felder) (Lepidoptera: Saturniidae,Hemileucinae)

Egg, larva, and pupa of Gamelia anableps (C. Felder & R. Felder, 1874) are described for the first time and the chaetotaxy of the first instar provided. Eggs and selected structures of larvae and pupae were also investigated by means of scanning electron microscopy. Eggs are laid in clusters, are ellipsoidal and laterally flat. Larvae passed through eight instars in about 78 days. Pupation occurs inside a poorly organized cocoon of yellowish silk. The dark brown pupa is cylindrical in shape with appendages firmly fused together and to the body wall. The chaeotaxy is compared with other Saturniidae species belongining to Hemileucinae, Ceratocampinae, and Saturniinae and the natural history of G. anableps compared with other Saturniidae.     

An adaptive bivariate kernel smoothing method for determining instars of Austrosimulium tillyardianum (Diptera: Simuliidae) larvae

1. In insects, instar determination is generally based on the frequency distribution of sclerotised body part measurements. Commonly used univariate methods, such as histograms and univariate kernel smoothing, are not sufficient to reflect the distribution of the measurements, because development of sclerotised body parts is multidimensional. 2. This study used an adaptive bivariate kernel smoothing method, based on 10 pairs of separating variables, to differentiate instars of Austrosimulium tillyardianum (Diptera: Simuliidae) larvae in two‐dimensional space. A variable bandwidth matrix was used and separation lines between instars were defined. Using the Crosby growth ratio, Brooks' rule and the new standard recently proposed, larvae were separated into nine instars. It was found that, using the bivariate kernel smoothing method, the clustering accuracy and determination of separation lines as instar class limits were higher than those associated with the univariate kernel smoothing method. With the exceptions of the paired separating variables, head capsule length and antennal segment 3 length (AS3L), the mean probabilities of correct classifications was > 85%. The pair of separating variables that yielded the greatest classification accuracy comprised mandible length (ML) and AS3L, which had mean probabilities of 0.8984. The clustering accuracy was higher for early‐ and late‐instar larvae, but lower for instars 6 and 7. The adaptive bivariate kernel smoothing method was better than univariate methods for instar determination, especially in the detection of divisions between instars and identification of a larval instar.     

Determining the number of instars in potato tuber moth Phthorimaea operculella (Zeller) using density‐based DBSCAN clustering

The potato tuber moth (PTM), Phthorimaea operculella (Zeller), is an important pest of Solanaceae crops and especially devastating to potatoes. There is no significant difference in morphological characteristics of PTM from the first to third instar larvae; therefore, it is difficult to directly determine the number of instars of this pest based on morphology. In the present study, head capsule width and length and mandible width of 340 PTM individuals were measured. Density‐based spatial clustering of applications with noise (DBSCAN) clustering was used for instar grouping. The results of DBSCAN clustering were compared with those obtained using Gaussian mixture models and k‐means clustering; the results of the three clustering methods were verified using Brooks–Dyar rule, Crosby rule and linear regression model. The clusters obtained using the three methods were the same and comprised four PTM instars with three morphological characteristics. Moreover, the results of the three methods fit the Brooks–Dyar rule, Crosby rule, frequency analysis and logarithmic regression model well. Head capsule width was the best morphological characteristic for determining the number of instars of PTM, and this characteristic may be used for determining PTM instars in the field. These results show that the DBSCAN clustering method is a promising tool for the identification of insect instars.     

Limb regeneration and apolysis in the tick, Ornithodoros tartakovskyi.

Limb regeneration potential and the apolysis process were investigated in the argasid tick, Ornithodoros tartakovskyi. Developmental instars received single or multiple amputations and were subsequently allowed to undergo single or multiple apolyses. Amputated ticks regenerated complete normal limbs but only after four successive apolyses. Following a single apolysis, the majority of regenerated limbs were essentially miniature duplicates of normal legs but commonly lacked normal chaetotaxy and/or tarsal hump(s). The site of amputation distal to the coxa-trochanter joint, number of limbs removed from an individual, and instar amputated did not consistently influence the extent of regeneration. Coagulation and clot formation were observed.The limbs of the tick apolysed within the old leg hulls. Larvae and nymphs amputated relatively early during the period of apolyses regenerated limbs; late amputations precluded regeneration. The process of apolysis was irreversible and not obviously affected by amputations.     

Analysis of time- and concentration-mortality relationship of nitenpyram against different larval stages of Nilaparvata lugens (Hemiptera: Delphacidae)

Bioassay of nitenpyram was conducted on the planthopper Nilaparvata lugens (St?l) (Hemiptera: Delphacidae) in the laboratory by using the rice-stem dipping method. The resulting data were analyzed using the complementary log-log time-concentration-mortality modeling technique, yielding the parameters for time and concentration effects of nitenpyram against N. lugens instars. The LC50 values to instars I and II estimated for 24, 48, and 72 h after treatment were 1.03, 0.27, and 0.11 mg/liter, respectively. These values were lower than those for instars III and IV and instars V at the same times. The estimates of LT50 for instars I and II were decreased from 69.06 to 16.22 h with the increasing concentration from 0.125 to 2 mg/liter; the same estimates for both instars III and IV and instars V at the concentrations of 1-2 mg/liter were close to each other, ranging from 33.11 to 44.16 h, much higher than the values of instars I and II at the same concentration. Based on the time-concentration-mortality relationships fitted and the virulence indices (LC50 and LT50) estimated, nitenpyram is a promising pesticide for N. lugens control.     

Argentophilic structures as a diagnostic criterion for the discrimination of species of the genus Gyrodactylus von Nordmann (Monogenea)

The mapping of argentophilic surface sensory structures (chaetotaxy) is a technique which has contributed to the taxonomy of larval digeneans. Four species of Gyrodactylus from four different hosts (Salmo salar, Gasterosteus aculeatus, Poecilia reticulata and Carassius auratus) were investigated using this technique in order to ascertain its usefulness in distinguishing species of this monogenean genus. A formula for the arrangement of the natural aggregations of sensilla similar to those prepared for cercariae is proposed. Although the greatest concentration of sensilla occur adjacent to the anterior extremity of the body, it is the region just posterior to the pharynx which most readily permits the discrimination of Gyrodactylus spp. based on the number and spatial configuration of these sensilla. Maps have been constructed to demonstrate these differences. It is concluded that chaetotaxy shows potential as a simple and reliable method of distinguishing species of this genus.     

A Novel Technique for Identifying the Instar of Field-Collected Insect Larvae

Many field studies of insects have focused on the adult stage alone, likely because immature stages are unknown in most insect species. Molecular species identification (e.g., DNA barcoding) has helped ascertain the immature stages of many insects, but larval developmental stages (instars) cannot be identified. The identification of the growth stages of collected individuals is indispensable from both ecological and taxonomic perspectives. Using a larval–adult body size relationship across species, I present a novel technique for identifying the instar of field-collected insect larvae that are identified by molecular species identification technique. This method is based on the assumption that classification functions derived from discriminant analyses, performed with larval instar as a response variable and adult and larval body sizes as explanatory variables, can be used to determine the instar of a given larval specimen that was not included in the original data set, even at the species level. This size relationship has been demonstrated in larval instars for many insects (Dyar’s rule), but no attempt has been made to include the adult stage. Analysis of a test data set derived from the beetle family Carabidae (Coleoptera) showed that classification functions obtained from data sets derived from related species had a correct classification rate of 81–100%. Given that no reliable method has been established to identify the instar of field-collected insect larvae, these values may have sufficient accuracy as an analytical method for field-collected samples. The chief advantage of this technique is that the instar can be identified even when only one specimen is available per species if classification functions are determined for groups to which the focal species belongs. Similar classification functions should be created for other insect groups. By using those functions together with molecular species identification, future studies could include larval stages as well as adults.     

Diagnosis of species by the larval phase in the genus Rhipicephalus Koch (Ixodidae) in USSR fauna

The key for identification of species of the genus Rhipicephalus (the fauna of the USSR) on the larval phase is based on the material reared under laboratory conditions. The structure of scutum, organs of gnathosoma and coxae, chaetotaxy of idiosoma and gnathosoma were used for differential diagnosis. In addition morphometric characteristics of the above structures and their proportions were used. A special attention has been given to coexistence of species in sympatric zones, which was revealed or confirmed as a result of identification of species of the genus on larva in the ranges of the fauna of the USSR.     

Hydaticus tuyuensis Trémouilles (Coleoptera: Dytiscidae): Larval Morphology and Phylogenetic Relationships within Dytiscinae

The three larval instars of Hydaticus (Guignotites) tuyuensis Trémouilles are described and illustrated for the first time, emphasizing the morphometry and chaetotaxy. Second- and third-instar larvae of the known species of the subgenus Guignotites Brinck are characterized by a trilobate median process of prementum. All larval instars of H. tuyuensis, however, have a bilobate process, similar to that present in the known species of the subgenus Hydaticus Leach. This implies that a trilobate process cannot be used as a diagnostic character for Guignotites. Alternatively, Guignotites as presently defined may not represent a natural group. A cladistic analysis of 57 larval characters suggests that the genus Hydaticus shares a common origin with the clade composed of the genera Eretes Laporte and Thermonectus Dejean, based on the following synapomorphies: (i) abdominal segment VII almost completely sclerotized ventrally; the absence of (ii) additional ventroapical pores on the third antennomere, (iii) setae FE4 and FE6, (iv) additional setae on the femur and (v) additional setae on the tibia; and the presence of (vi) setae on the median process of prementum and (vii) spinulae on the second labial palpomere. A bilobate or trilobate median process of the prementum and the submedial insertion of seta AN3 distinguish Hydaticus from the remaining genera of Dytiscinae studied.     

Scanning electron microscopy of the four larval instars of the Dengue fever vector Aedes aegypti (Diptera: Culicidae)

Aedes aegypti is the main insect vector of Dengue fever and dengue hemorrhagic fever/dengue shock syndrome and represents the only vulnerable element in the control of this disease. Therefore, the identification and quantification of this mosquito is an important task; however, the majority of taxonomic keys are based on the 4th larval instar. For that reason, this study describes the four larval instars ofA. aegypti using scanning electron microscopy. Morphological changes during larval development were observed at the pecten, comb scales and the ventral brush of the abdominal segment X; however, the 3rd and 4th instars showed similar structures with only a slight variation. The structures described in this study will be helpful in the identification of the four instars of A. aegypti, a fundamental task for comprehending the natural history of dengue mainly in new territories affected.     

Cercarial chaetotaxy of Codonocephalus urniger (Rudolphi, 1819) (Trematoda: Diplostomidae)

The chaetotaxy of Codonocephalus urniger (Rudolphi, 1819) cercariae recovered from Lymnaea palustris in a brackish water lake in Bulgaria is described and figured. Comparisons with Diplostomum spathaceum (Rudolphi, 1819) and D. pseudospathaceum Niewiadomska, 1984 cercariae indicate a similar general pattern in the distribution of sensilla but with marked differences in their number and arrangement on the ventral sucker, tail stem and furcae. Data on the chaetotaxy of other diplostomid species are used to reveal the similarities and variations in some complexes within the members of Diplostomidae with known cercarial chaetotaxy.     

Why the simple frequency and Janetschek methods are unreliable for determining instars of mayflies: An analysis of the number of Nymphal instars of stenonema modestum (banks) (ephemeroptera: Heptageniidae) in virginia,USA

The Janetschek method is a variation of the simple frequency method and therefore subject to the same limitations. Instars can only be reliably determined by these methods if development within the population is known to be homogeneous. The number of nymphal instars proposed by Kondratieff and Voshell (1980) for an overwintering population of Stenonema modestum in the North Anna River of Virginia is not substantiated. These authors estimated a total of 14–15 nymphal instars using the simple frequency and Janetschek methods. A reexamination of their paper indicates that the development of this 5. modestum population is quite variable and therefore not amenable to successful instar determination using the simple frequency, Janetschek or any other indirect instar determination method. Indirect instar determination methods cannot be considered reliable in determining the number of instars of Ephemeroptera because of the developmental variability characteristic of most, if not all, mayfly species.