A Multilocus Genotypic Analysis of Cryptosporidium meleagridis

ABSTRACT. Cryptosporidium meleagridis is a common cause of cryptosporidiosis in birds. In addition, recent reports have described the parasite as an etiologic agent of cryptosporidiosis in both immunocompetent and immunocompromised humans. Therefore, it is important to genetically characterize isolates of C. meleagridis from different hosts and geographic areas, and to develop molecular tools to differentiate isolates from various hosts or areas. In this study, a total of 11 isolates of Cryptosporidium meleagridis from both human and avian hosts were examined at three genetic loci: the small-subunit rRNA, 60-kDa glycoprotein precursor, and 70-kDa heat shock protein genes. Two genotypes of C. meleagridis were seen at the small-subunit rRNA locus. These differed from each other by the presence or lack of a heterogeneous copy of the gene and an ATT repeat. The 60-kDa glycoprotein precursor gene divided these eleven isolates of C. meleagridis into six genotypes with high sequence diversity between groups. The highest genetic heterogeneity, however, was seen at the 70-kDa heat shock protein locus, and was primarily present at the 3'end of the gene. This heterogeneity separated eight isolates of C. meleagridis into six genotypes. These data could be useful in the development of molecular tools to promote understanding of the transmission of C. meleagridisi in humans.     

Adipokinetic hormone counteracts oxidative stress elicited in insects by hydrogen peroxide: in vivo and in vitro study

The role of adipokinetic hormone (AKH) in counteracting oxidative stress elicited in the insect body is studied in response to exogenously applied hydrogen peroxide, an important metabolite of oxidative processes. In vivo experiments reveal that the injection of hydrogen peroxide (8 µmol) into the haemocoel of the firebug, Pyrrhocoris apterus L. (Heteroptera: Pyrrhocoridae) increases the level of AKH by 2.8‐fold in the central nervous system (CNS) and by 3.8‐fold in the haemolymph. The injection of hydrogen peroxide also increases the mortality of experimental insects, whereas co‐injection of hydrogen peroxide with Pyrap‐AKH (40 pmol) reduces mortality to almost control levels. Importantly, an increase in haemolymph protein carbonyl levels (i.e. an oxidative stress biomarker) elicited by hydrogen peroxide is decreased by 3.6‐fold to control levels when hydrogen peroxide is co‐injected with Pyrap‐AKH. Similar results are obtained using in vitro experiments. Oxidative stress biomarkers such as malondialdehyde and protein carbonyls are significantly enhanced upon exposure of the isolated CNS to hydrogen peroxide in vitro, whereas co‐treatment of the CNS with hydrogen peroxide and Pyrap‐AKH reduces levels significantly. Moreover, a marked decrease in catalase activity compared with controls is recorded when the CNS is incubated with hydrogen peroxide. Incubation of the CNS with hydrogen peroxide and Pyrap‐AKH together curbs the negative effect on catalase activity. Taken together, the results of the present study provide strong support for the recently published data on the feedback regulation between oxidative stressors and AKH action, and implicate AKH in counteracting oxidative stress. The in vitro experiments should facilitate research on the mode of action of AKH in relation to oxidative stress, and could help clarify the key pathways involved in this process.     

The germarium of panoistic ovarioles of Bacillus rossius (Insecta Phasmatodea): Structure and function during imaginal life

Summary

In adult females of Bacillus rossius (Insecta Phasmatodea) the germarium, localized at the ovariole tip just below the terminal filament and above the vitellarium, progressively reduces in size and eventually disappears at the end of the ovulatory period. The observations with light and electron microscopes show that in the end-chamber most germ cells are arrested in a post-pachytenic diffuse stage, which just precedes diplotenic oocyte growth. These observations also indicate that the reduction in size of the germarium of ovulating females should probably be ascribed to a progressive and extensive activation of the resting germ cells. The average number of ovulated eggs per ovariole (6.7±0.9) is consistent with this view. However, occasional findings of lepto-zygotenic germ cells in some preovulatory ovarioles of adult females do not completely rule out the persistence of scarce undifferentiated germ elements (oogonia) in the larval germarium at the onset of adult life. Furthermore, the reduction of the germarium in ovulating females and its subsequent disappearance in post-ovulating ones also includes the somatic cells, which are always present among the germ cells in previous stages. Since each early growing oocyte becomes surrounded by a thin monolayer of follicle cells, the diminution of end-chamber somatic cells supports the view that they actually represent prefollicular cells, which are progressively utilized from the onset of imaginal life onwards.     

Tasphorura, a new genus of Tullbergiinae (Collembola: Onychiuridae) abundant in Tasmanian Nothofagus forest

Abstract. Tasphorura , a new genus of onychiurid Collembola, is erected for an unusually coloured species, Tasphorura vesiculata n.sp., found at a single locality in Tasmanian Nothofagus rainforest. Its isolated phyletic position is noted and extremely high densities are recorded. The conservation implications of the restricted distribution shown by this species, which is typical of many rainforest invertebrates, are discussed with reference to increasing utilization and fragmentation of these forests.     

Testing the regeneration niche hypothesis with Gesneriaceae (tribe Sinningiae) in Brazil: Implications for the conservation of rare species

The germination requirements of sexually reproducing plants are regulated by environmental factors such as temperature. Those factors acting at the germination phase are part of the regeneration niche, which is fundamental in the processes that contribute to habitat suitability and geographic distribution. We tested the hypothesis that rarity is associated with regeneration niche in three species of plants in the family Gesneriaceae (tribe Sinningieae), Sinningia rupicola (Mart.) Wiehler, Paliavana sericiflora Benth and Sinningia allagophylla (Mart.) Wiehler, which vary in their distribution and habitat specificity but share a small zone of sympatry in rocky fields south of Belo Horizonte in Minas Gerais, Brazil. The regeneration niche was tested using a seed germination experiment under controlled light conditions at seven fixed temperatures (10–40°C at 5°C intervals). Each of the three species germinated differently at the various temperatures. The species with the smallest geographic range, S. rupicola, also had the most restricted germination: germination peaked at 15°C when relatively few seeds germinated (45%), and even fewer germinated at other temperatures. The regeneration niche was wider in P. sericiflora and wider still in S. allagophylla, with germination greater than 90% between 15–25°C and greater than 80% between 15–30°C, respectively. Our germination results provide qualified support for the hypothesis of correlation of the regeneration niche with geographic distribution of related plant taxa, with important conservation implications for rare and endangered species.     

Local host ant specificity of Phengaris (Maculinea) teleius butterfly,an obligatory social parasite of Myrmica ants

1. Phengaris butterflies are obligatory social parasites of Myrmica ants. Early research suggested that there is a different Myrmica host species for each of the five European Phengaris social parasites, but more recent studies have shown that this was an oversimplification. 2. The pattern of host ant specificity within a Phengaris teleius metapopulation from southern Poland is reported. A combination of studying the frequency distribution of Phengaris occurrence and morphometrics on adult butterflies were used to test whether use of different host species is reflected in larval development. 3. Phengaris teleius larvae were found to survive in colonies of four Myrmica species: M. scabrinodis, M. rubra, M. ruginodis, and M. rugulosa. Myrmica scabrinodis was the most abundant species under the host plant but the percentage of infested nests was similar to other host ant species at two sites and lower in comparison to nests of M. rubra and M. ruginodis at the other two sites. Morphometric measurements of adult butterflies reared by wild colonies of M. scabrinodis and M. ruginodis showed that wing size and number of wing spots were slightly greater for adults eclosing from nests of M. ruginodis. 4. Our results suggest that P. teleius in the populations studied is less specialised than previously suggested. The results are consistent with the hypothesis that P. teleius is expected to be the least specific of the European Phengaris species, as it has the largest and best defended fourth‐instar caterpillars and, as a predatory species, it spends less time in the central larval chambers of the host colonies. The fact that individuals reared by M. ruginodis had wider hind wings may suggest that P. teleius had better access to resources in M. ruginodis than in M. scabrinodis colonies.     

Artificial intelligence in pest insect monitoring

Abstract Global problems of hunger and malnutrition induced us to introduce a new tool for semi‐automated pest insect identification and monitoring: an artificial neural network system. Multilayer perceptrons, an artificial intelligence method, seem to be efficient for this purpose. We evaluated 101 European economically important thrips (Thysanoptera) species: extrapolation of the verification test data indicated 95% reliability at least for some taxa analysed. Mainly quantitative morphometric characters, such as head, clavus, wing, ovipositor length and width, formed the input variable computation set in a Trajan neural network simulator. The technique may be combined with digital image analysis.     

Conflicting patterns of genetic and morphological variation in European Gentianella section Gentianella

Genetics in European Gentianella Moench sect. Gentianella were investigated using internal transcribed spacer (ITS) nrDNA, trn L-F cpDNA and Amplified Fragment Length Polymorphisms (AFLP) fingerprint data to analyse relationships within the section. Both a cladistic and a phenetic approach, as well as nonhierarchical analysis (PCA) and partitioning of molecular variance (AMOVA), were used. Sequence data did not resolve groups within the section. They reflected, however, geographical isolation between central European and eastern Carpathian ( G. lutescens and G. bulgarica ) as well as Caucasian samples ( G. caucasea ). AFLP data revealed a strong differentiation within G. germanica between populations of the Alps and north-western middle Europe, although there is hardly any morphological differentiation of taxonomic significance. Morphological similarities between G. aspera and G. pilosa were not reflected in genetics, and no support was found for G. aspera . Genetic similarities were not in line with morphology in G. lutescens and G. bulgarica . AFLP data only partly resolve relationships among taxa. High genetic similarities, as well as partitioning of molecular variance among taxa/populations of eastern middle Europe ( G. austriaca , G. bohemica , G. germanica , G. stiriaca ) and intermediate morphological characters in G. bohemica and G. stiriaca , might indicate reticulate evolution. The overall low sequence divergence within the section points to a very young origin of all taxa.  © 2005 The Linnean Society of London, Botanical Journal of the Linnean Society , 2005, 148 , 175–187.