Morphological and histological study of eggs of six species of the Triatoma genus (Hemiptera: Reduviidae)

The objective of this study was to analyze the morphological and histological characteristics of eggs from six triatomine species in order to obtain a generic and specific characterization of this group of Trypanosoma cruzi vectors and better understand their phylogenetic and taxonomic aspects. The eggs of the respective species came from the collection of the Laboratório de Triatomíneos e Culicídeos da Faculdade de Saúde Pública/USP, from the Insetário do Servi?o Especial de Saúde de Araraquara - SESA and from the Triatomine insectary of the Laboratório Nacional e Internacional de Referência em Taxonomia de Triatomíneos (FIOCRUZ), in Rio de Janeiro. Morphological studies were done with optical microscopy and scanning electron microscopy. Histological analyses used only optical microscopy. All of the species analyzed showed a predominance of hexagonal cells. The exchorion of Triatoma breyeri Del Ponte, T. costalimai Verano Galv?o, and T. tibiamaculata (Pinto) consists of unornamented cells, overlapping or slightly separated, with a smooth, padded appearance. Meanwhile, in T. matogrossensis Leite Barbosa, T. sherlocki Papa, Juberg, Carcavallo, Cerqueira Barata, and T. williami Galv?o, Souza Lima, the exchorion cells are ornamented on their entire surface with perforations and fissures. Egg histology showed undulations, pores, and orifices. The study helps to expand the generic and specific knowledge of the Triatominae subfamily. The characteristics identified in the exchorion of the eggshells can help separate close species. These new parameters will back the elaboration of future dichotomous keys, help to determine each vector species role and expand knowledge of the various species.     

Numerical analyses of seed morphology and its taxonomic significance in the tribe Nigelleae (Ranunculaceae)

We examined the seed morphology of Nigella, Garidella and Komaroffia to aid systematics and taxonomy. The seeds of 21 taxa in 20 species from Mediterranean and West Asian countries were studied using both zoom stereo and scanning electron microscopy (SEM). Principal coordinate analysis (PCoA) and unweighted average linkage analysis (UPGMA) were performed to assess the contribution of seed characters to the taxonomy of the tribe. Representative pictures from zoom stereo microscopy and SEM studies and measurement of seeds are presented. Most studied species showed unique seed surface sculpturing features, and seed shape, size and surface sculpturing separated the studied taxa into five main groups. However, the variation in seed shape and color were found to be in conflict with the generic taxonomy while macromorphological characters were found more congruent with the sectional taxonomy than with the generic taxonomy. Five main types of seed ornamentations – rugulose + striate, granulate, smooth, rugulose and mucronulate – were identified. Patterns in seed micromorphology strongly supported the sectional level taxonomy of Nigella. Two of the three sections of the genus were found to be well differentiated in terms of seed characters. Furthermore, seed morphology alone provide good support to Garidella as a distinct genus, but shows less taxonomic value for Komaroffia.     

Ability of bed bug-detecting canines to locate live bed bugs and viable bed bug eggs

The bed bug, Cimex lectularius L., like other bed bug species, is difficult to visually locate because it is cryptic. Detector dogs are useful for locating bed bugs because they use olfaction rather than vision. Dogs were trained to detect the bed bug (as few as one adult male or female) and viable bed bug eggs (five, collected 5-6 d after feeding) by using a modified food and verbal reward system. Their efficacy was tested with bed bugs and viable bed bug eggs placed in vented polyvinyl chloride containers. Dogs were able to discriminate bed bugs from Camponotus floridanus Buckley, Blattella germanica (L.), and Reticulitermes flavipes (Kollar), with a 97.5% positive indication rate (correct indication of bed bugs when present) and 0% false positives (incorrect indication of bed bugs when not present). Dogs also were able to discriminate live bed bugs and viable bed bug eggs from dead bed bugs, cast skins, and feces, with a 95% positive indication rate and a 3% false positive rate on bed bug feces. In a controlled experiment in hotel rooms, dogs were 98% accurate in locating live bed bugs. A pseudoscent prepared from pentane extraction of bed bugs was recognized by trained dogs as bed bug scent (100% indication). The pseudoscent could be used to facilitate detector dog training and quality assurance programs. If trained properly, dogs can be used effectively to locate live bed bugs and viable bed bug eggs.     

北京猛禽两种旋尾类寄生线虫的报道(线虫纲,旋尾目)(英文)

报道寄生于北京猛禽的2种旋尾类线虫,其中一种为新种,另一种为中国新纪录种。作者分别对其做了光镜和电镜观察。北京副旋翼线虫,新种Paraspiralatus beijingensis sp.nov.采自于雕鸮的口腔和胃内,它与P. sakeri Gibbons,Nicholls &Bailey,2004的不同之处在于具有单个的肛前乳突,右交合刺远末端弯向背侧并稍微膨大而P. sakeri的交合刺远末端是直的,不膨大,呈钝尖形。纵纹腹小鸮无肛线虫Aprocta noctuae Spaul,1928采自红角鸮的体腔。本次所观察标本在形态上,如交合刺的长短和形状,尾乳突的数目和排列,阴门的位置,卵的大小等,都与前人对A. noctuae的描述一致。但是电子扫描电镜结果显示,A. noctuae的头端具有8个头乳突而不是4个,线虫角质层具有精细的横纹。A. noctuae为我国新纪录种。标本保存于河北师范大学生命科学学院。     

Scanning electron microscopy of the eggs of Ascaris lumbricoides, A. suum, Toxocara canis, and T. mystax.

Eggs of Ascaris lumbricoides, A. suum, Toxocara canis, and T. mystax were examined by scanning electron microscopy (SEM). All species under study exhibited pronounced surface ridges. The ridges formed distinctive patterns in T. canis and T. mystax. In the Ascaris species, the ridges are similar except that they are more pronounced in the eggs of A. suum. Operculumlike structures were observed only in Ascaris. Correlation of data from SEM with previously reported transmission electron microscopy suggests that the surface ridges seen in Ascaris eggs are formed by the chitinous layer of the shell.     

Contribution to the taxonomy of the family Campulidae Odhner, 1926 (Digenea) by means of a morphometric multivariate analysis

Digeneans of the family Campulidae occur exclusively in marine mammals, particularly in cetaceans. Their taxonomy is confused, being based on adult morphology only. We used a multivariate discriminant analysis of morphometric data to provide new evidence on the taxonomy of the Campulidae. Measurements of 217 specimens from 21 species of all seven genera of the family were taken. The percentage of specimens correctly assigned into their own species was 96.3%. The first three discriminant functions accounted for most of the variation between the species, which were grouped together in suprageneric groups along the first and the second function. The ordination pattern observed conforms partly with the established taxonomy of the Campulidae. Variation along the first discriminant function can be interpreted as a gradient of the vitelline extension and the maximum width to length ratio. This function separates three groups of species, corresponding to the Hunterotrematinae, Orthosplanchninae and Campulinae-Lecithodesminae. The last two subfamilies are separated along the second axis, the Lecithodesminae having longer bodies and suckers than the Campulinae. This study also provides some evidence at the generic and specific level.     

Respiratory morphology of the Abedus herberti Hidalgo egg chorion (Hemiptera: Belostomatidae)

Although giant water bugs (Hemiptera: Belostomatidae) are large, aquatic insects known for their obligate paternal egg brooding behaviors, little research has focused on the structure of their eggs. The respiratory requirements of developing embryos likely created selection for brooding, so a thorough understanding of the respiratory morphology of belostomatid eggs could help explain how brooding behaviors facilitate embryonic gas exchange. This study used scanning electron microscopy to document the respiratory microstructure of the eggs of Abedus herberti, a back brooding giant water bug. The exochorion is similar to that of other belostomatids in texture and organization except that the respiratory region is confined to the uppermost quarter of the egg. This is the area exposed to the atmosphere by encumbered males. A plastron network made up of densely packed vertical projections demarcates the boundary between the respiratory and nonrespiratory regions of the chorion. The internal chorion is composed of alternate air‐filled and denser layers that likely facilitate the movement of oxygen from the aeropyles at the top of the eggs to the developing embryonic tissues. J. Morphol., 2011. © 2011 Wiley‐Liss, Inc.     

Health food versus fast food: the effects of prey quality and mobility on prey selection by a generalist predator and indirect interactions among prey species

1. In order to understand the relative importance of prey quality and mobility in indirect interactions among alternative prey that are mediated by a shared natural enemy, the nutritional quality of two common prey for a generalist insect predator along with the predator's relative preference for these prey was determined. 2. Eggs of the corn earworm Helicoverpa zea (Lepidoptera: Noctuidae) were nutritionally superior to pea aphids Acyrthosiphum pisum (Homoptera: Aphididae) as prey for big‐eyed bugs Geocoris punctipes (Heteroptera: Geocoridae). Big‐eyed bugs survived four times as long when fed corn earworm eggs than when fed pea aphids. Furthermore, only big‐eyed bugs fed corn earworm eggs completed development and reached adulthood. 3. In two separate choice experiments, however, big‐eyed bugs consistently attacked the nutritionally inferior prey, pea aphids, more frequently than the nutritionally superior prey, corn earworm eggs. 4. Prey mobility, not prey nutritional quality, seems to be the most important criterion used by big‐eyed bugs to select prey. Big‐eyed bugs attacked mobile aphids preferentially when given a choice between mobile and immobilised aphids. 5. Prey behaviour also mediated indirect interactions between these two prey species. The presence of mobile pea aphids as alternative prey benefited corn earworms indirectly by reducing the consumption of corn earworm eggs by big‐eyed bugs. The presence of immobilised pea aphids, however, did not benefit corn earworms indirectly because the consumption of corn earworm eggs by big‐eyed bugs was not reduced when they were present. 6. These results suggest that the prey preferences of generalist insect predators mediate indirect interactions among prey species and ultimately affect the population dynamics of the predator and prey species. Understanding the prey preferences of generalist insect predators is essential to predict accurately the efficacy of these insects as biological control agents.     

Isoenzymatic characterization of potentially useful forage shrubs belonging to the genus Cytisus and allies (Genistea: Leguminosae)

Twenty-five accessions belonging to the genus Cytisus and allies (Genisteae: Leguminosae), potentially useful for agrosylvopastoral systems in the Mediterranean basin, were analyzed using isoenzymes. The aim was to assess the taxonomic usefulness of these characters at the specific and generic levels, and its value as an aid for the taxonomy. A high level of infrageneric and infraspecific variability was detected. Numerical analysis showed that isoenzymes are useful for the characterization both at the specific and generic levels. The obtained dendrogram supports the generic arrangement proposed by Bisby [(1981) Advances in Legume Systematics, Vol. 1, pp. 409–425], therefore helping to elucidate the difficult taxonomy of the studied species.     

Chorion patterns on eggs of Lutzomyia sandflies from the Peruvian Andes

Abstract. Scanning electron microscopy (SEM) was used to depict and describe the eggs of five species of phlebotomine sandfly from the Andean region of Peru: Lutzomyia caballeroi, Lu.noguchii, Lu.peruensis, Lu.tejadai and Lu.verrucarum. Two new types of chorionic sculpturing of sandfly eggs are reported: these were named disperse (Lu.tejadai) and verrucose (Lu.noguchii). The aeropylar area of the eggs is also described for the first time for neotropical sandflies. These character states appear to vary for ecological rather than phylogenetic reasons and could be used for species identification.     

Spore morphology as a taxonomic tool in the delimitation of three Asplenium L. species complexes (Aspleniaceae: Pteridophyta) in Cuba

As a contribution to the taxonomic study of Asplenium L. for the new "Flora de la República de Cuba", the spore morphology was studied into three complexes of hardly delimited species. Twenty-two collections have been studied by SEM and eighty-nine by LM. Shape, number of spores per sporangium and perispore ornamentation, specially fine details revealed by scanning electron microscopy, were the most valuable characters in the recognition of taxa. Irregularities in spore shape and size, unusual development of perispore wall surface elements, and collapsed protoplast characterize aborted spores and suggest the occurrence of a possible hybrid species.     

Tuyamaella serratistipa S. Hatt. new to the Philippines,with a key to the genus Tuyamaella (Hepaticae,Lejeuneaceae)

Abstract

Tuyamaella serratistipa S. Hatt. is reported as a new generic and specific record for the Philippine bryoflora. A detailed illustration of the species and a distribution map of the genus and the species are presented. A key to all known species of Tuyamaella is also provided, and the taxonomy of the genus discussed.     

Biological characteristics of the mirids Macrolophus costalis and Macrolophus pygmaeus preying on the tobacco form of Myzus persicae (Hemiptera: Aphididae)

Biological and population parameters of the predatory bugs Macrolophus costalis Fieber and Macrolophus pygmaeus Rambur preying on the tobacco aphid Myzus persicae (Sulzer) were examined. Tobacco was used as host plant and all experiments were carried out at 23 degrees C and L16:D8. In M. costalis, the developmental time for eggs and the total duration of nymphal instars was 13.1 and 22.2 days and in M. pygmaeus 13.0 and 20.2 days, respectively. Both species completed their nymphal development feeding only on plant juices although they required significantly more time to attain adulthood than when feeding on prey. The longevity of M. costalis females and the preoviposition period were 49.9 and 5.6 days and those of M. pygmaeus were 50.3 and 6.3 days, respectively. Adult males showed a higher longevity than females in both species. The intrinsic rate of increase and the mean total number of eggs laid were 0.0644 and 121 in M. costalis and 0.0615 and 104 in M. pygmaeus, respectively. Mean total aphid consumption by nymphs of M. costalis was 61 aphids, about twice that of M. pygmaeus (37 aphids). Total aphid consumption by a male and female pair of M. costalis and M. pygmaeus until the death of the female was 244 and 285 aphids, respectively. The results of the study are discussed in relation to the impact of the two predatory bugs on tobacco aphid populations.     

The relationship between predator density, community composition, and field predation of Colorado potato beetle eggs

Conservation biological control programs seek to increase natural enemy densities through the adoption of more benign farming practices, under the assumption that higher predator densities will lead to more effective pest suppression. However, predator–predator interference may lead to diminishing returns in improved pest control as predator densities increase. We examined the relationship between predator density and predation rates on Colorado potato beetle eggs in production potato fields. These potato fields naturally spanned a 10-fold range in predator density, due to differences in management practices. Periodically through the growing season we simultaneously measured predator densities and subjected sentinel eggs masses to predation, allowing us to correlate predator density and egg predation for each field on each sample date. Egg predation rates were significantly positively correlated with total predator densities, a correlation that was not improved when predator densities were scaled to reflect differences in feeding rates on potato beetle eggs of the constituent predator taxa. There was no correlation between per-capita egg predation rates and predator density, and so no evidence that predator interference increased with increasing predator density. We divided predators into six dominant taxa—dwarf spiders, crab spiders, minute pirate bugs, big-eyed bugs, damsel bugs, and Lygus bugs (together constituting 93% of all predators collected), and a seventh group, “other predators” that included all other, less common, taxa—and examined correlations between all predator combinations and egg predation rates. The highest correlation was between combined densities of the six most common predator taxa, excluding only the “other predators” grouping. This suggests that predators may be largely equivalent in their impact on Colorado potato beetle eggs, and that field scouts might be able to ignore uncommon predator taxa when sampling for natural enemies.     

SYSTEMATIC IMPLICATIONS OF SEED COAT MORPHOLOGY IN NAMA (HYDROPHYLLACEAE)

Seeds from 37 species of Nama (Hydrophyllaceae) were examined by scanning electron microscopy in order to assess systematic implications of seed coat structure. Generally, seed coat morphology is species specific. Nevertheless, similarities among species in seed coat organization, particularly, outer testa anatomy and wall thickening-pitting patterns allow the recognition of six groups among examined taxa; furthermore, seed coat features often suggest both intra-group and inter-group relationships. Recognized groupings do not correspond well with the more classicial treatments of Nama; rather, they suggest that a restructuring of the infrageneric and even the generic taxonomy of Nama is in order.     

Gut symbiotic bacteria of the genus Burkholderia in the broad-headed bugs Riptortus clavatus and Leptocorisa chinensis (Heteroptera: Alydidae)

The Japanese common broad-headed bugs Riptortus clavatus and Leptocorisa chinensis possess a number of crypts in the posterior region of the midgut, whose lumen contains a copious amount of bacterial cells. We characterized the gut symbiotic bacteria by using molecular phylogenetic analysis, light and electron microscopy, in situ hybridization, and PCR-based detection techniques. Restriction fragment length polymorphism analysis of 16S rRNA gene clones suggested that a single bacterium dominated the microbiota in the crypts of the both bug species. The predominant 16S rRNA gene sequences obtained from different individuals and species of the bugs were not identical but were very similar to each other. Homology searches in the DNA databases revealed that the sequences showed the highest levels of similarity (96% to 99%) to the sequences of Burkholderia spp. belonging to the beta subdivision of the class Proteobacteria. In situ hybridization with specific oligonucleotide probes confirmed the localization of the Burkholderia symbiont in the lumen of the midgut crypts. Electron microscopy showed that the lumen of the crypts was filled with rod-shaped bacteria of a single morphotype. Molecular phylogenetic analysis demonstrated that the Burkholderia symbionts of the bugs formed a well-defined monophyletic group, although the group also contained several environmental Burkholderia strains. The phylogenetic relationship of the Burkholderia symbionts did not reflect the relationship of the host bug species at all. The sequences from R. clavatus and the sequences from L. chinensis did not form clades but were intermingled in the phylogeny, suggesting that horizontal transmission of the symbiont might have occasionally occurred between populations and species of the bugs.     

Integrative taxonomy,or iterative taxonomy?

The recently introduced term ‘integrative taxonomy’ refers to taxonomy that integrates all available data sources to frame species limits. We survey current taxonomic methods available to delimit species that integrate a variety of data, including molecular and morphological characters. A literature review of empirical studies using the term ‘integrative taxonomy’ assessed the kinds of data being used to frame species limits, and methods of integration. Almost all studies are qualitative and comparative – we are a long way from a repeatable, quantitative method of truly ‘integrative taxonomy’. The usual methods for integrating data in phylogenetic and population genetic paradigms are not appropriate for integrative taxonomy, either because of the diverse range of data used or because of the special challenges that arise when working at the species/population boundary. We identify two challenges that, if met, will facilitate the development of a more complete toolkit and a more robust research programme in integrative taxonomy using species tree approaches. We propose the term ‘iterative taxonomy’ for current practice that treats species boundaries as hypotheses to be tested with new evidence. A search for biological or evolutionary explanations for discordant evidence can be used to distinguish between competing species boundary hypotheses. We identify two recent empirical examples that use the process of iterative taxonomy.     

Characterization of an obligate intracellular bacterium in the midgut epithelium of the bulrush bug Chilacis typhae (Heteroptera, Lygaeidae, Artheneinae)

Many members of the suborder Heteroptera have symbiotic bacteria, which are usually found extracellularly in specific sacs or tubular outgrowths of the midgut or intracellularly in mycetomes. In this study, we describe the second molecular characterization of a symbiotic bacterium in a monophagous, seed-sucking stink bug of the family Lygaeidae (sensu stricto). Chilacis typhae possesses at the end of the first section of the midgut a structure which is composed of circularly arranged, strongly enlarged midgut epithelial cells. It is filled with an intracellular endosymbiont. This "mycetocytic belt" might represent an evolutionarily intermediate stage of the usual symbiotic structures found in stink bugs. Phylogenetic analysis based on the 16S rRNA and the groEL genes showed that the bacterium belongs to the Gammaproteobacteria, and it revealed a phylogenetic relationship with a secondary bacterial endosymbiont of Cimex lectularius and free-living plant pathogens such as Pectobacterium and Dickeya. The distribution and ultrastructure of the rod-shaped Chilacis endosymbiont were studied in adults and nymph stages using fluorescence in situ hybridization (FISH) and electron microscopy. The detection of symbionts at the anterior poles of developing eggs indicates that endosymbionts are transmitted vertically. A new genus and species name, "Candidatus Rohrkolberia cinguli," is proposed for this newly characterized clade of symbiotic bacteria.     

Immature stages of Meiosimyza Hendel 1925 and related genera (Diptera,Lauxaniidae)

The immature stages of Holometabolan insects can provide valuable data both for taxonomy and phylogeny, but they are well known only for a negligible proportion of the described species. Scanning electron microscopy (SEM) analysis of all preimaginal stages of the lauxaniid species Meiosimyza decempunctata (Fallén 1820), M. platycephala (Loew 1847), M. rorida (Fallén 1820), M.?subfasciata (Zetterstedt 1838), Tricholauxania praeusta (Fallén 1820) and Aulogastromyia anisodactyla (Loew 1845) is presented for the first time. Detailed images of eggs and all three larval instars are provided along with illustrations of the cephaloskeleton and photos of puparia. The egg is rather simple, white, with developed longitudinal ridges. The body integument of all larval instars is smooth and uniform across species. The cephaloskeleton is slender; the mouth hooks are parallel, subhypostomal sclerites are arched and fusing to a ventral bridge. The puparium is brownish, oval, without calcareous deposits. Generally, the morphology of larvae and puparia is very conservative. On the other hand, the eggs show considerable degree of interspecific variability.     

Breeding ecology and seasonal abundance of the giant water bug Appasus japonicus (Heteroptera, Belostomatidae)

Males of the giant water bug Appasus (= Diplonychus) japonicus Vuillefroy (Belostomatidae: Heteroptera) carry egg masses on their back, but little is known about the relationship between seasonal abundance and breeding ecology of the species. In the present study, therefore, a field survey based on a mark-and-recapture census was carried out at three survey points within a rice paddy area (0.3 km²) where A. japonicus forms a meta-population in northern Okayama, Japan. We investigated the body size, seasonal abundance, dispersion, egg mass size (number of eggs within one egg mass), number of egg masses and the total eggs carried on the back of each male as fundamental parameters of the population and breeding. Significant differences in egg mass size, number of egg masses, and total number of eggs that males carried was found among the survey points. The present results suggested the possibility that the differences in breeding parameters of A. japonicus were influenced by differences in environmental factors among the microhabitats. These results are discussed in conjunction with previous reports on seasonal abundance and breeding systems in Belostomatidae bugs.