A new microsporidian pathogen, Vairimorpha gastrophysae sp. nov., isolated from Gastrophysa viridula (Coleoptera: Chrysomelidae).

Gastrophysa viridula DeGeer 1775, the green dock leaf beetle, belongs to a group of beneficial insects, which can be used as a classical biological control agent against sorrels (Rumex sp., Polygonaceae). Therefore, any infection by pathogenic organisms in this beetle is undesirable. In the present study, a new microsporidian pathogen isolated from G. viridula was identified based on morphological and ultrastructural characteristics, supported with a molecular phylogenetic analysis. Light and transmission electron microscopy studies indicated that the microsporidium was polymorphic throughout its life cycle. Sporulation stages were not all in direct contact with the host-cell cytoplasm. The fresh single diplokaryotic spores of the secondary sporulation cycle had a long narrow morphology, measuring about 5 × 2.1 µm (n = 50). Octospores produced in the secondary sporulation cycle were also observed. Morphological and ultrastructural characteristics of the life cycle stages place it within the genus Vairimorpha. The phylogenetic tree constructed on the 16S rRNA gene sequence analysis supports the morphological and ultrastructural characteristics and indicates that the pathogen is closely related to the Vairimorpha clade of microsporidia. The pathogen is named Vairimorpha gastrophysae sp. nov.

Early Screening for Posttraumatic Stress Disorder in Inpatient Detoxification and Motivation Treatment: Results and Consequences.

AIMS: Posttraumatic stress disorder (PTSD) is a significant comorbidity in substance use disorders (SUDs). While most studies have addressed trauma/PTSD in abstinent patients, little is known about trauma/PTSD in early detoxification treatment. The current study therefore addresses the systematic evaluation of trauma/PTSD in early inpatient detoxification. METHODS: A cross-sectional survey was accomplished in three German-speaking clinics (n = 134) specialized in inpatient detoxification and motivation treatment. All measures are based on self-report using trauma-specific questionnaires and measures for general psychopathological burden. RESULTS: Participation rate was 60.1% and patients did not show clinically obvious psychological distress during or after assessment. DSM-IV traumatic events were reported by 66.4%. Of the total sample, 38.1% screened positive for PTSD, and 14.9% screened positive for subsyndromal PTSD. PTSD patients reported significantly more childhood adversities and significantly higher scores in depression and -general psychopathology compared to subsyndromal PTSD and SUD-only patients. CONCLUSIONS: Early and systematic evaluation of PTSD in SUD inpatient detoxification treatment is largely safe and yields important information for individual treatment. The high PTSD-rate and the high symptom load in SUD patients during inpatient detoxification treatment highlight the need for a more stringent address of trauma/PTSD in early SUD treatment.

Natural occurrence of microsporidia infecting Lepidoptera in Bulgaria.

We examined 34 lepidopteran species belonging to 12 families to determine presence and prevalence of microsporidian pathogens. The insects were collected from May 2009 to July 2012 from 44 sites in Bulgaria. Nosema species were isolated from Archips xylosteana, Tortrix viridana, Operophtera brumata, Orthosia cerasi, and Orthosia cruda. Endoreticulatus sp. was isolated from Eilema complana. The prevalence of all isolates in their hosts was low and ranged from 1.0% to 5.3%. Phylogenetic analyses of the new isolates based on SSU rDNA are presented.

First report and spore ultrastructure of Vairimorpha plodiae (Opisthokonta: Microspora) from Plodia interpunctella (Lepidoptera: Pyralidae) in Turkey.

The present study describes the first isolation and characterization of Vairimorpha plodiae, a microsporidian pathogen of Plodia interpunctella (Lepidoptera: Pyralidae), from Turkey. We present characteristic light and electron microscopical features of the spores. Fresh binucleate spores are oval and measure 4.48 ± 0.23 (4.01-4.84) µm in length and 2.21 ± 0.15 (1.91-2.48) µm in width. Ultrastructural studies showed that the spore wall measures 150 to 200 nm and consists of a clear endospore (125-150 nm) and an electron-dense, uniform, thin exospore (30-50 nm). The polar filament is isofilar and with 10-12 coils. The well-developed polaroplast consists of two parts with thin lamellae anteriorly and thick, irregularly arranged lamellae posteriorly. The identity of our isolate is discussed.

Stability of trauma-related symptoms during acute substance use treatment.

It is unclear whether post-traumatic stress disorder symptoms and reports of traumatic childhood experiences decline during substance withdrawal. A convenience sample of 34 inpatients of the Psychiatric University Clinics in Basel was recruited and general psychopathological and trauma-related symptoms were assessed with the Brief Symptom Checklist, Post-Traumatic Stress Diagnostic Scale, and Childhood Trauma Questionnaire in the 1st and 3rd week of substance use treatment. The average age of the sample was 41.9 (SD = 9.1) years, and 26.5% were female. Hyperarousal (Mt1 = 4.51 versus Mt2 = 3.61; z = -2.38, p = .017) and avoidance symptoms (Mt1 = 6.24 versus Mt2 = 4.27; z = -2.59, p = .010) declined significantly, but re-experiencing symptoms (Mt1 = 4.00 versus Mt2 = 3.45; z = -.50, p = .617) did not. Post-traumatic stress disorder assessment, according to the Diagnostic and Statistical Manual of Mental Disorders-4th edition criteria, remained constant for 28 of 34 patients. Likewise, self-reported childhood trauma experiences decreased, yet the number of elevated subscale scores remained stable. Post-traumatic stress disorder symptoms are not adequately treated by substance withdrawal alone. Trauma-specific diagnostics can be initiated with sufficient quality as early as the first week of withdrawal treatment.

Review of the genus Endoreticulatus (Microsporidia, Encephalitozoonidae) with description of a new species isolated from the grasshopper Poecilimon thoracicus (Orthoptera: Tettigoniidae) and transfer of Microsporidium itiiti Malone to the genus.

The historic genus Pleistophora (Plistophora) is a highly polyphyletic clade with invertebrate Microsporidia reassigned to several new genera since the 1980s. Two genera, Endoreticulatus and Cystosporogenes, clearly separate into distinct but closely related clades based on small subunit ribosomal RNA analysis but are included in different families that are each polyphyletic. A microsporidium with morphology resembling the Endoreticulatus/Cystosporogenes clade was isolated from the grasshopper Poecilimon thoracicus from a site in Northwest Bulgaria. It produced intense infections in the digestive tract of the host but no behavioral changes were noted in infected individuals. Prevalence of the microsporidium increased over the active feeding season yearly. Mature spores were oval and measured 2.58±0.21 µm×1.34±0.24 µm, with 16 to approximately 32 spores in a parasitophorous vacuole. The spores were uninucleate and polar filament coils numbered 8-9 situated in a single row. The spore polaroplast consisted of an anterior lamellar section and a posterior vesicular section, and the posterior vacuole was reduced. Analyses of a 1221 bp partial SSU-rRNA sequence indicated that the isolate is more closely related to the Endoreticulatus clade than to Cystosporogenes, but shows earlier phylogenetic separation from species infecting Lepidoptera and represents a new species, Endoreticulatus poecilimonae. To compare sequences of Endoreticulatus spp. from Lepidoptera to those infecting other insect orders, an isolate, Microsporidium itiitiMalone (1985), described from the Argentine stem weevil, Listronotus bonariensis, was sequenced. Like the grasshopper isolate, the weevil isolate is closely related but basal to the lepidopteran Endoreticulatus clade. The original description combined with the new sequence data confirms species status and permits transfer of the isolate from Microsporidium, a genus erected for microsporidian species of uncertain taxonomic status, to Endoreticulatus.

The first record of nucleopolyhedrovirus isolated from the gypsy moth, Lymantria dispar (Lepidoptera, Lymantriidae) in Turkey.

OBJECTIVE: The gypsy moth, Lymantria dispar L. (Lepidoptera, Lymantriidae) is a common pest of forests and fruit trees throughout the world. This insect is also a major serious pest in Turkey. Nowadays L. dispar can be managed by biological control methods especially, using entomopathogenic viruses. The aim of this study is to characterize entomopathogenic viruses and is the first record of nucleopolyhedrovirus isolated from the gypsy moth in Turkey. METHODS: PIBs obtained from infected larvae were measured and photographed using an Olympus BX51 microscope with a DP-25 digital camera and a DP2-BSW Soft Imaging System and examined with a Philips 208 electron microscope (TEM). RESULTS: The virus had the typical characteristics of nucleopolyhedroviruses. The dimension of the polyhedral inclusion bodies (PIBs) was 2.03±0.25 µm. PIBs varied in size from 1.65 to 2.21 µm and were usually polygonal in shape. Virions in PIBs contained 1 to 8 nucleocapsids per virion. The size of the viral particles was 366.67±54.72 (312-500) x 42.95±6.12 (30-47) nm. CONCLUSION: The isolation and characterization of a pure isolate of Lymantria dispar multinucleopolyhedrovirus (LdMNPV-TR) from Turkey is presented for the first time.

A cell culture model for Nosema ceranae and Nosema apis allows new insights into the life cycle of these important honey bee-pathogenic microsporidia.

The population of managed honey bees has been dramatically declining in the recent past in many regions of the world. Consensus now seems to be that pathogens and parasites (e.g. the ectoparasitic mite Varroa destructor, the microsporidium Nosema ceranae and viruses) play a major role in this demise. However, little is known about host-pathogen interactions for bee pathogens and attempts to develop novel strategies to combat bee diseases have been hampered by this gap in our knowledge. One reason for this dire situation is the complete lack of cell cultures for the propagation and study of bee pathogens. Here we present a cell culture model for two honey bee-pathogenic microsporidian species, Nosema apis and N. ceranae. Our cell culture system is based on a lepidopteran cell line, which proved to be susceptible to infection by both N. ceranae and N. apis and enabled us to illustrate the entire life cycle of these microsporidia. We observed hitherto undescribed spindle-shaped meronts and confirmed our findings in infected bees. Our cell culture model provides a previously unavailable means to explore the nature of interactions between the honey bee and its pathogen complex at a mechanistic level and will allow the development of novel treatment strategies.

Five-year cohort study of Nosema spp. in Germany: does climate shape virulence and assertiveness of Nosema ceranae?

Nosema ceranae and Nosema apis are two fungal pathogens belonging to the phylum Microsporidia and infecting the European honeybee, Apis mellifera. Recent studies have suggested that N. ceranae is more virulent than N. apis both at the individual insect level and at the colony level. Severe colony losses could be attributed to N. ceranae infections, and an unusual form of nosemosis is caused by this pathogen. In the present study, data from a 5-year cohort study of the prevalence of Nosema spp. in Germany, involving about 220 honeybee colonies and a total of 1,997 samples collected from these colonies each spring and autumn and analyzed via species-specific PCR-restriction fragment length polymorphism (RFLP), are described. Statistical analysis of the data revealed no relation between colony mortality and detectable levels of infection with N. ceranae or N. apis. In addition, N. apis is still more prevalent than N. ceranae in the cohort of the German bee population that was analyzed. A possible explanation for these findings could be the marked decrease in spore germination that was observed after even a short exposure to low temperatures (+4 degrees C) for N. ceranae only. Reduced or inhibited N. ceranae spore germination at low temperatures should hamper the infectivity and spread of this pathogen in climatic regions characterized by a rather cold winter season.

Horizontal and vertical transmission of a Nosema sp. (Microsporidia) from Lymantria dispar (L.) (Lepidoptera: Lymantriidae).

The gypsy moth, Lymantria dispar L. (Lepidoptera, Lymantriidae), a serious defoliator of deciduous trees, is an economically important pest when population densities are high. Outbreaking populations are, however, subject to some moderating influences in the form of entomopathogens, including several species of microsporidia. In this study, we conducted laboratory experiments to investigate the transmission of an unusual Nosema sp. isolated from L. dispar in Schweinfurt, Germany; this isolate infects only the silk glands and, to a lesser extent, Malpighian tubules of the larval host. The latent period ended between 8 and 15 days after oral inoculation and spores were continuously released in the feces of infected larvae until pupation. Exclusion of feces from the rearing cages resulted in a 58% decrease in horizontal transmission. The silk of only 2 of 25 infected larvae contained microsporidian spores. When larvae were exposed to silk that was artificially contaminated with Nosema sp., 5% became infected. No evidence was found for venereal or transovum (including transovarial) transmission of this parasite.

[Chronic fatigue syndrome--a functional somatic syndrome]. / Chronisches Erschöpfungssyndrom--eine funktionelle Störung.

Chronic fatigue can be categorized as a functional somatic syndrome (fss), because there are findings of typical preconditions, trigger mechanisms and maintaining conditions. With relevance for therapy it makes sense to see it as an medical-psychiatric interface-disorder Subsyndromal short episodes of chronic fatigue are many more frequent as three or six month during clearly diagnosed episodes of "neurasthenia" or "chronic fatigue syndrome". Their descriptions are very similar and obvious those means the same matter. For original aetiological assumptions it wasn't any evidence. But there are findings of charcteristical patterns of changed neurhumeral and immunological interactions for the chronic fatigue syndrome, common for fss. Especially changes of HPA-Axis and its interactions with other systems of functional regulation. Another importent fact are increased senzitation in neuronal and neurocognitive regulation. Increased critical appraisal of somatic funtions and dysfunctional coping strategies are maintaining factors at least. Patterns of dysfuntional coping are not a problem of patients alone. There are also experiences, that some doctors shows the same dysfunctional somatizing management of fss in general and especially for chronic fatigue. In fact, a single and specific cause of chronic fatigue doesn't exist. But the above-mentioned facts allows a starting point for a more successful treatment. There are reviews that shows a good evidence for therapeutic procedures wich are calling for acticvity by patients, such cognitive behavioral therapy and graduated activation. Antidepressants, especialy SSRI, are helpful with a small evidence. They can be used to increase treatment effects. There is no evidence for therapies without patients activation.

Vairimorpha disparis n. comb. (Microsporidia: Burenellidae): a redescription and taxonomic revision of Thelohania disparis Timofejeva 1956, a microsporidian parasite of the gypsy moth Lymantria dispar (L.) (Lepidoptera: Lymantriidae).

Investigation of pathogens of populations of the gypsy moth, Lymantria dispar (L.) in Central and Eastern Europe revealed the existence of a microsporidium (Fungi: Microsporidia) of the genus Vairimorpha. The parasite produced three spore morphotypes. Internally infective spores are formed in the gut and adjacent muscle and connective tissue; single diplokaryotic spores and monokaryotic spores grouped by eight in sporophorous vesicles develop in the fat body tissues. The small subunit rDNA gene sequences of various isolates of the Vairimorpha microsporidia, obtained from L. dispar in various habitats in the investigated region, revealed their mutual identity. In phylogenetic analyses, the organism clustered with other L. dispar microsporidia that form only diplokaryotic spores in the sporogony cycle. The octospores of certain microsporidia infecting Lepidoptera that were previously described as Thelohania spp., have recently been shown to be one of the several spore morphotypes produced by species in the genus Vairimorpha. Because the description and drawings of a parasite described as Thelohania disparis by Timofejeva fit the characteristics of Vairimorpha, and all octospore-producing microsporidia collected from L. dispar since 1985 are genetically identical Vairimorpha species, it is believed that the parasite characterized here is identical to T. disparis Timofejeva 1956, and is herein redescribed, characterized, and transferred to the genus Vairimorpha as the new combination Vairimorpha disparis n. comb.

Microsporidian infections in Lymantria dispar larvae: interactions and effects of multiple species infections on pathogen horizontal transmission.

The interactions in multiple species infections and effects on the horizontal transmission of three microsporidian species, Vairimorpha disparis, Nosema lymantriae and Endoreticulatus schubergi, infecting Lymantria dispar were evaluated in the laboratory. Simultaneous and sequential inoculations of host larvae were performed and the resulting infections were evaluated. Test larvae were exposed to the inoculated larvae to measure horizontal transmission. Dual species infections demonstrated interspecific competition between Nosema and Vairimorpha in the host larvae, but no observable competition occurred between Endoreticulatus and either of the other microsporidian species. Timing of inoculation was an important factor determining the outcome of competition between Nosema and Vairimorpha. The species inoculated first showed a higher rate of successful establishment; a time lag of 7 days between inoculations allowed the first species to essentially exclude the second. The microsporidia differed in efficiency of horizontal transmission. Nosema and Endoreticulatus were transmitted at very high rates, close to 100%. Horizontal transmission of Vairimorpha was less efficient, ranging from 25% to a maximum of 75%. The patterns of infection observed in inoculated larvae were reflected in the test larvae that acquired infections in the horizontal transmission experiments. Competition with Vairimorpha suppressed horizontal transmission of Nosema after simultaneous and sequential inoculation. In simultaneous inoculation experiments Endoreticulatus had no effect on transmission of Nosema and Vairimorpha.

Nosema chrysorrhoeae n. sp. (Microsporidia), isolated from browntail moth (Euproctis chrysorrhoea L.) (Lepidoptera, Lymantriidae) in Bulgaria: characterization and phylogenetic relationships.

A new microsporidian parasite Nosema chrysorrhoeae n. sp., isolated in Bulgaria from the browntail moth (Euproctis chrysorrhoea L.), is described. Its life cycle includes two sequential developmental cycles that are similar to the general developmental cycles of the Nosema-like microsporidia and are indistinguishable from those of two Nosema spp. from Lymantria dispar. The primary cycle takes place in the midgut tissues and produces binucleate primary spores. The secondary developmental cycle takes place exclusively in the silk glands and produces binucleate environmental spores. N. chrysorrhoeae is specific to the browntail moth. Phylogenetic analysis based on the ssu rRNA gene sequence places N. chrysorrhoeae in the Nosema/Vairimorpha clade, with the microsporidia from lymantriid and hymenopteran hosts. Partial sequences of the lsu rRNA gene and ITS of related species Nosema kovacevici (Purrini K., Weiser J., 1975. Natürliche Feinde des Goldafters, Euproctis chrysorrhoea L., im Gebiet von Kosovo, FSR Jugoslawien. Anzeiger fuer Schädlingskunde, Pflanzen-Umweltschutz, 48, 11-12), Nosema serbica Weiser, 1963 and Nosema sp. from Lymantria monacha was obtained and compared with N. chrysorrhoeae. The molecular data indicate the necessity of future taxonomic reevaluation of the genera Nosema and Vairimorpha.

Studies on the impact of two Nosema isolates from Bulgaria on the gypsy moth (Lymantria dispar L.).

We investigated host-parasite interactions of two Nosema-type microsporidian isolates recovered from populations of Lymantria dispar L. in northwestern Bulgaria, one near Veslec and one near Levishte. Bioassay studies produced information on development, stage specific mortality, pupation, and adult eclosion of infected individuals. Horizontal transmission of the two isolates was investigated in a second set of experiments. At dosages ranging from 2 x 10(2) to 5 x 10(4) spores/microl, the infection rates varied between 77 and 100% for the isolate from Veslec and between 92 and 99% for the Levishte isolate. The Veslec isolate caused a slightly higher mortality rate and the median time to death was shorter compared to the isolate from Levishte. The total mortality for both isolates varied between 79 and 99%, independent of spore dosages. A lower relative growth rate was recorded for male and female L. dispar larvae infected with either isolate during the third larval instar and a higher relative growth rate during the fourth instar compared to the control groups. Pupal weight did not differ significantly among females, but male infected pupae were heavier than the controls. Nosema sp. [Veslec] was as efficiently transmitted as Nosema sp. [Levishte]; 42% of the susceptible larvae became infected with the Veslec isolate when uninfected larvae were exposed to infected larvae; 43% of larvae became infected with the Nosema sp. [Levishte]. The latency period varied between 7 and 8 days for both isolates.