Search

Thyridanthrax fenestratus (Fallén, 1814) is a bombyliid with poorly understood biology. It was recently shown to locally but frequently parasitize Pemphredon fabricii (M. Müller, 1911) (Hymenoptera: Crabronidae), a crabronid wasp that abundantly nests in old Lipara-induced galls on the common reed Phragmites australis (Cav.) Trin. ex Steud., 1840. The parasitism modes in Bombyliidae and Thyridanthrax spp. are not uniform. Here, we report that Th. fenestratus switches facultatively between killing the host almost immediately (idiobiont strategy) and killing the host at a later developmental stage (koinobiont strategy). We document the koinobiont parasitoid strategy for a series of Th. fenestratus larvae parasitizing P. fabricii. We found that a significant portion of Th. fenestratus larvae spend winter as young larvae and start feeding on fully developed and defecated prepupae of P. fabricii only after the end of cold-induced winter diapause. The time needed for the development of Th. fenestratus larvae exceeds several times the time needed for pupation of P. fabricii prepupae; the parasitized prepupae, therefore, remain paralyzed until the parasitic larva completes feeding. Fungicides, which alter the pupation of the host larva, seem to have negligible effects on Th. fenestratus larvae. The ability to switch between the two parasitism strategies has already been reported for several Anthrax spp., though the ability to block the host in the defecated prepupa stage and prevent its pupation following cold-induced diapause is herein reported for the first time.

Avian Dicrocoeliidae are difficult to identify, even in their adult stages. Molecular analyses have been conducted for only a few species, complicating the accurate identification of juvenile stages. The taxonomy of the family is unresolved, and the status of many dicrocoeliid species is uncertain. Sequences of nuclear and mitochondrial DNA loci of Central European avian Dicrocoeliidae were generated and analyzed. These included representatives of the genera Lyperosomum, Platynosomum, Stromitrema, Brachylecithum, Brachydistomum, and Lutztrema. All the sequences were obtained from morphologically identified adult specimens of dicrocoeliids isolated from avian hosts. Molecular support was obtained to validate Lyperosomum turdia, confirm the rejection of Lyperosomum dujardini and Lyperosomum alagesi, and resurrect Lyperosomum longicauda and Lyperosomum collurionis. The identity of European Platynosomum illiciens from avian hosts with American vouchers of the same species from avian and mammalian hosts was confirmed. Brachylecithum fringillae is not considered valid; the individuals that matched its diagnosis were subadult Brachydistomum ventricosum. Descriptions and comparative data for five new species are provided. These are Lyperosomum hirundinis sp. n., Lyperosomum tenori sp. n., Lyperosomum atricapillae sp. n., Stromitrema acrocephali sp. n., and Lutztrema atricapillae sp. n.. Based on the molecular data, suggestions are provided regarding the validity of dicrocoeliid species that parasitize Central European birds. Further research should address the polyphyletic status of Brachylecithum.

Abstract Ants are key ecosystem service providers and can serve as important biological control agents in pest management. However, the effects of insecticides on common farmland ant species are poorly understood. We tested the effects of three commonly used insecticides on ants (Hymenoptera, Formicidae). The tested insecticides were acetamiprid (neonicotinoid; formulated as Mospilan 20 SP), deltamethrin (pyrethroid; formulated as Sanium Ultra), and sulfoxaflor (sulfilimine; formulated as Gondola). We tested two ant (Hymenoptera: Formicidae) species with different colony founding strategies, Lasius niger (Linnaeus, 1758) and Myrmica rubra (Linnaeus, 1758). We sprayed their queens with insecticides at concentrations recommended for use in foliar applications in agriculture, i.e., at 1.25 g L−1 (acetamiprid), 0.6 g L−1 (sulfoxaflor), and 0.875 g L−1 (deltamethrin). Further, we diluted the compounds in distilled water and tested them at 10%, 1%, and 0.1% of the field-recommended concentrations, and used distilled water as a control. We monitored the survival of the queens and the number of eggs laid. All three tested insecticides caused severe lethal and sublethal concentration-dependent effects. Even at concentrations three orders of magnitudes lower than recommended for field applications, significantly lower numbers of eggs were found in the queens’ nests. The extent of the sublethal effects of acetamiprid and sulfoxaflor was concentration-dependent and differed between the two ant species. Besides bees and bumblebees, ants represent an important group of hymenopterans that are severely affected even by low concentrations of the tested compounds and therefore should be included in risk assessment schemes.

Abstract The diversity, host specificity, and physiological effects of endosymbiotic bacteria in spiders (Araneae) are poorly characterized. We used 16S rDNA sequencing to evaluate endosymbionts in the cephalothorax and legs of a wolf spider Pardosa agrestis. We tested the effects of feeding once or twice daily with fruit flies, aphids, or starved and compared them to those of syntopically occurring Pardosa palustris. The feeding increased traveled distance up to five times in some of the groups provisioned with food relative to the starved control. The Shannon diversity t-test revealed significant differences between these component communities of the two spider species. The increased frequency of feeding with fruit flies, but not aphids, increased the dominance and decreased the alpha diversity of OTUs. The obligate or facultative endosymbionts were present in all analyzed spider individuals and were represented mostly by Rickettsiella, Rhabdochlamydia, Spiroplasma, and the facultative intracellular parasite Legionella. Vertically transmitted endosymbionts were less common, represented by Wolbachia pipientis and Rickettsia sp. H820. The relative abundance of Mycoplasma spp. was negatively correlated with provisioned or killed aphids. In conclusion, the tissues of Pardosa spiders host tremendously diverse assemblages of bacteria, including obligate or facultative endosymbionts, with yet unknown phenotypic effects.

Abstract Background Central European aerial insectivores are long-distance migrants that winter in sub-Saharan Africa. Most of them employ the fly-and-forage migrating strategy and differ in their food composition. The composition and structure of helminth component communities of these hosts are poorly understood, and information regarding seasonality and long-term changes is unavailable. Methods From 1963 to 2022, we analyzed the population trends of helminths in five aerial insectivore species. Namely, we examined Apus apus, Hirundo rustica, Delichon urbicum, Riparia riparia, and Ficedula albicollis; all originated from the Czech Republic. Results We identified central European aerial insectivores as hosts that are parasitized mostly by helminths that cannot complete their life-cycles in the nesting quarters of their hosts. This phenomenon is unknown in other bird host species. In contrast, only a single dominant trematode species that completes its life-cycle locally colonized the central European aerial insectivores. All other dominant species of Trematoda, all Nematoda, and all Acanthocephala were dependent on intermediate hosts unavailable in the nesting quarters of the examined bird hosts. Surprisingly, these helminths transmitted from winter quarters or migratory routes were diverse, and many of them were abundant in terms of both prevalence and intensity of infection. The helminth component communities of aerial insectivores were dynamic systems. During the study period, three species became new and regularly encountered members of helminth fauna of examined hosts, and other species gradually increased or decreased their intensity of infection. In contrast to other groups of bird hosts, the dominant helminth species of aerial insectivores did not experience local extinctions or rapid population losses. Conclusions The analysis of helminths of five central European aerial insectivores revealed component communities that heavily rely on completing host–parasite cycles at migration routes or wintering grounds. The composition of the analyzed component communities changed dynamically during the 60-year-long study period, but there was no evidence of large-scale declines in abundance or prevalence. Graphical Abstract

Morphological stasis is a widespread characteristic of pseudoscorpions, suggesting that much cryptic diversity remains unexplored. Here, we revise the polytypic species Dactylochelifer latreillii in the framework of an integrative taxonomic approach, using DNA barcoding, multivariate ratio analysis, geometric morphometry of the male foretarsus, and genitalic morphology. The pattern of mitochondrial variation suggests three species-level entities in central Europe, which widely overlap in morphospace, but differentiate in the structure of the female genitalia, and by their ecology. Dactylochelifer latreillii (Leach) is a halobiont species, occurring exclusively in coastal habitats and in Pannonian salt steppes, and D. l. septentrionalis Beier syn. nov. is a junior synonym of the nominate species. Dactylochelifer degeerii (C. L. Koch) stat. rev. is the oldest available name for an inland species that has long been mistaken for the nominotypical subspecies of D. latreillii. New habitat information suggests a preference for higher shrub vegetation. Dactylochelifer ninnii (Canestrinii) stat. rev. is a halophilic Mediterranean species that extends to the northern limits of the Pannonian basin. The distinctiveness of the Mediterranean “form” was recognized by early naturalists in the 19th century, but was ignored by later authorities in the field.

The spatial distribution of parasitoids is closely linked to the distribution and ecological requirements of their hosts. Several studies have documented changes in the fauna composition of parasitoids in response to elevation, but data on parasitoids associated with spiders are missing. The koinobiont ichneumonid wasp Zatypota anomala is strictly specialised on spiders of the genus Dictyna (Dictynidae) in Europe. We examined the distribution of spiders of the family Dictynidae in forest ecotones in central Europe across a broad elevation gradient (110–1466 m a.s.l.). We checked the spiders for parasitism by Z. anomala. It was most abundant at the mid-elevations (median 712 m a.s.l., range 179–870 m a.s.l.). We identified four dictynid spider species as Z. anomala hosts. These were Dictyna arundinacea, Dictyna uncinata, Nigma flavescens, and Nigma walckenaeri. All four species and the genus Nigma were recorded as hosts for the first time. The parasitoids strongly preferred juvenile instars of their hosts. The body length differed between parasitised Dictyna and Nigma spiders (medians: 1.95 mm and 2.55 mm, respectively). The distribution of Dictyna and Nigma spiders overlapped along the elevation gradient, but parasitism incidence significantly differed between spider genera along the elevation gradient. Nigma was parasitized at lower elevations between 179–254 m a.s.l. and Dictyna at higher elevations between 361–870 m a.s.l. The phenology of Z. anomala is closely tied to the univoltine life strategy of its host spiders. The parasitoid female oviposits in autumn, and its offspring overwinter as larvae on the host, reach adulthood during spring, and pass the summer as an adult.

The bee genus Pseudoanthidium is represented by nine species in Europe. Of these nine species, Pseudoanthidium nanum is the most widespread, occurs mainly in xerothermic open habitats and creates nests in various cavity types. In this study, we provide information on the nest structure of this species in reed stalks and oak galls and about its parasitic species. We provide the first report of P. nanum as a host of Xylophrurus augustus (Ichneumonidae). The biology of the much rarer related species Pseudoanthidium tenellum is described here for the first time. This species occurs in terrestrial reed beds and wet meadows with the presence of reed galls and flowering plants in the family Asteraceae and is rare throughout its entire distribution area. This species nests inside reed galls induced by Lipara frit flies, and the nest structure is very similar to that of P. nanum. We report new parasitic species of this bee, namely, the cuckoo bee Stelis punctulatissima, the predator-inquiline Gasteruption nigrescens and two parasitoids, Leucospis biguetina and Miltogramma punctata. This bee collects pollen mainly from wetland plants in Bidens and Pulicaria. We also describe mature larvae of both species. The larvae do not differ greatly from one another; only the shape of mandibles and sclerotisation of mouthparts are slightly different. Further research should address the ecological requirements of P. tenellum, a poorly understood reed gall inquiline.

Abstract Exposure to numerous chemicals disrupts the spiders' locomotion. Spiders, particularly epigeic spiders, are dependent on their locomotory activities to search for prey, hide from their enemies, and perform sexual reproduction and subsequent parental care. Among the best-known compounds that inhibit the locomotion of arthropods are neonicotinoids. Despite spiders are less affected by the neonicotinoids than insects due to the sequence differences in their acetylcholine receptors, they are not resistant to these compounds. We hypothesized that acute exposure to a broad spectrum of neonicotinoids suppresses the traveled distance, mean velocity, and maximum velocity in epigeic spiders. As a model species, we used adults of Pardosa lugubris. We tested commercial formulations of thiamethoxam, acetamiprid, and thiacloprid. We tested each of the neonicotinoids in the maximum and minimum concentrations recommended for foliar applications. We applied them under controlled conditions dorsally by spraying them directly on the spiders or exposing the spiders to the tarsal contact with neonicotinoid residues. Control groups consisted of 31 individuals; treated groups consisted of 10–21 individuals. We found that a broad spectrum of neonicotinoids temporarily suppresses the traveled distance in epigeic spiders. At 1 h after application, all the three tested neonicotinoid insecticides induced declines in the traveled distance, but this effect mostly disappeared when tested at 24 h after the application. The decrease in the traveled distance was associated with substantial temporary decreases in the mean and maximum velocities. Despite differences among modalities, all three insecticides caused multiple adverse effects on the locomotory parameters in any tested concentrations. It remains to test what would be the lowest safe concentration for the chronic exposure to neonicotinoids in epigeic spiders.

Abstract Background Parasitic conjunctivitis caused by Philophthalmus spp. is a common ophthalmic disease in birds, with localized outbreaks occurring worldwide. There is no consensus on treating this disease; mechanical removal is considered a standard recommendation, but is associated with disease relapses within days or weeks. Methods From 2015 to 2020, we examined 4295 Larus michahellis and Larus fuscus gulls in southern Portugal for the presence of Philophthalmus spp. Due to the need to treat dozens of infected gulls in the rescue station, we tested three treatment regimens aimed at targeting Philophthalmus lucipetus in the infected gulls: (I) the ophthalmic application of levamisole; (II) the oral application of milbemycin in combination with praziquantel; and (III) the subcutaneous application of ivermectin. Results The outbreak of philophthalmosis in gulls in southern Portugal has been ongoing since the first cases were reported in 2015–2016. The prevalence of philophthalmosis has fluctuated annually, peaking a maximum of 10.3% in L. fuscus in 2017 and at 2.1% in L. michahellis in 2016. The infection intensity peaked at a median of 11.5 eye-flukes per host bird in L. fuscus in 2016 and a median of six eye-flukes per host bird in L. michahellis in 2017. Nine gulls were infected with >50 eye-flukes. None of the treatment options were effective at treating P. lucipetus infections: the numbers of eye-flukes in the infected birds did not decrease, and the clinical signs of the disease did not change. Conclusions An outbreak of philophthalmosis in southern Portugal has massively affected two species of gulls in the region. Two previously suggested philophthalmosis treatments (ocular levamisole and praziquantel given orally), as well as a third mode of treatment with a previously failed compound (ivermectin administered subcutaneously) were used. However, the treatments did not affect the numbers of P. lucipetus in the eyes of the treated gulls. Further research should address ophthalmic gel formulations or sub-conjunctival delivery mode for antihelminthic drugs that are effective against Philophthalmus spp. in vitro. Graphical abstract

Protein tyrosine phosphatase, nonreceptor type 22 (PTPN22), is an archetypal non-HLA autoimmunity gene. It is one of the most prominent genetic contributors to type 1 diabetes mellitus outside the HLA region, and prevalence of its risk variants is subject to enormous geographic variability. Here, we address the genetic background of patients with type 1 diabetes mellitus of Armenian descent. Armenia has a population that has been genetically isolated for 3000 years. We hypothesized that two PTPN22 polymorphisms, rs2476601 and rs1310182, are associated with type 1 diabetes mellitus in persons of Armenian descent. In this association study, we genotyped the allelic frequencies of two risk-associated PTPN22 variants in 96 patients with type 1 diabetes mellitus and 100 controls of Armenian descent. We subsequently examined the associations of PTPN22 variants with the manifestation of type 1 diabetes mellitus and its clinical characteristics. We found that the rs2476601 minor allele (c.1858T) frequency in the control population was very low (q = 0.015), and the trend toward increased frequency of c.1858CT heterozygotes among patients with type 1 diabetes mellitus was not significant (OR 3.34, 95% CI 0.88-12.75; χ2 test p > 0.05). The control population had a high frequency of the minor allele of rs1310182 (q = 0.375). The frequency of c.2054-852TC heterozygotes was significantly higher among the patients with type 1 diabetes mellitus (OR 2.39, 95% CI 1.35-4.24; χ2 test p < 0.001), as was the frequency of the T allele (OR 4.82, 95% CI 2.38-9.76; χ2 test p < 0.001). The rs2476601 c.1858CT genotype and the T allele correlated negatively with the insulin dose needed three to six months after diagnosis. The rs1310182 c.2054-852CC genotype was positively associated with higher HbA1c at diagnosis and 12 months after diagnosis. We have provided the first information on diabetes-associated polymorphisms in PTPN22 in a genetically isolated Armenian population. We found only a limited contribution of the prototypic gain-of-function PTPN22 polymorphism rs2476601. In contrast, we found an unexpectedly close association of type 1 diabetes mellitus with rs1310182.

Abstract Background Hexokinases (HKs) are well-studied enzymes catalyzing the first step of glycolysis. However, non-canonical regulatory roles of HKs are still incompletely understood. Here, we hypothesized that HKs comprise one of the missing links between high-dose metformin and the inhibition of the respiratory chain in cancer. Methods We tested the isoenzyme-specific regulatory roles of HKs in ovarian cancer cells by examining the effects of the deletions of HK1 and HK2 in TOV-112D ovarian adenocarcinoma cells. We reverted these effects by re-introducing wild-type HK1 and HK2, and we compared the HK1 revertant with the knock-in of catalytically dead HK1 p.D656A. We subjected these cells to a battery of metabolic and proliferation assays and targeted GC×GC-MS metabolomics. Results We found that the HK1 depletion (but not the HK2 depletion) sensitized ovarian cancer cells to high-dose metformin during glucose starvation. We confirmed that this newly uncovered role of HK1 is glycolysis-independent by the introduction of the catalytically dead HK1. The expression of catalytically dead HK1 stimulated similar changes in levels of TCA intermediates, aspartate and cysteine, and in glutamate as were induced by the HK2 deletion. In contrast, HK1 deletion increased the levels of branched amino acids; this effect was completely eliminated by the expression of catalytically dead HK1. Furthermore, HK1 revertants but not HK2 revertants caused a strong increase of NADPH/NADP ratios independently on the presence of glucose or metformin. The HK1 deletion (but not HK2 deletion) suppressed the growth of xenotransplanted ovarian cancer cells and nearly abolished the tumor growth when the mice were fed the glucose-free diet. Conclusions We provided the evidence that HK1 is involved in the so far unknown glycolysis-independent HK1–metformin axis and influences metabolism even in glucose-free conditions.

Abstract Background The biodiversity of farmland habitats is witnessing unprecedented change, mostly in declines and simplification of assemblages that were established during centuries of the use of traditional agricultural techniques. In Central Europe, conspicuous changes are evident in populations of common farmland birds, in strong contrast to forest birds in the same region. However, there is a lack of information on longitudinal changes in trematodes that are associated with common farmland birds, despite the fact that diversity of trematodes is directly linked to the preservation of long-established food webs and habitat use adaptations of their hosts. Methods We analyzed the population trends of trematodes for the period 1963–2020 in six bird species that use Central European farmlands as their predominant feeding habitats. Namely, we examined Falco tinnunculus, Vanellus vanellus, winter populations of Buteo buteo, Ciconia ciconia, extravilan population of Pica pica, and Asio otus, all originating from the Czech Republic. Results We observed dramatic population losses of all trematode species in C. ciconia and V. vanellus; the changes were less prominent in the other examined hosts. Importantly, the declines in prevalence and intensity of infection affected all previously dominant species. These included Tylodelphys excavata and Chaunocephalus ferox in C. ciconia, Lyperosomum petiolatum in P. pica, Strigea strigis in A. otus, Neodiplostomum attenuatum and Strigea falconis in B. buteo (χ 2 test P

Abstract Spiders were recently shown to be adversely affected by field-realistic concentrations of a broad scale of neonicotinoid insecticides. Among the reported effects of neonicotinoids on invertebrates were declines in lipid biosynthesis and upregulation of β-oxidation, while vertebrate models suggest increased adipogenesis following treatment with neonicotinoids. Therefore, we hypothesized that there exists synergy between the effects of diet and concurrent exposure to field-realistic concentrations of neonicotinoid insecticides. To address this hypothesis, we fed first instars of the large wolf spider Hogna antelucana with two types of diets and exposed them to field-realistic concentrations of three formulations of neonicotinoids (thiamethoxam, thiacloprid and acetamiprid). We then measured the growth of the tested spiders; the lipid and protein content of their bodies; and their behavior, including ballooning, rappelling, and locomotor parameters. The two tested diets consisted of casein-treated and sucrose-treated Drosophila melanogaster. The dietary treatments affected the lipid and protein content of the spiders, their body weight and carapace length but did not affect any of the measured behavioral parameters. Surprisingly, we did not find any effects of acute exposure to neonicotinoid insecticides on the lipid or protein reserves of spiders. Exposure to neonicotinoids altered the behavior of the spiders as reported previously in other spider species; however, these effects were not affected by dietary treatments. Overall, the dietary treatments did not have any major synergy with acute exposure to field-realistic concentrations of neonicotinoid insecticides.

The mygalomorph spiders of the family Atypidae are among the most archaic spiders. The genus Atypus Latreille, 1804 occurs in Eurasia and northern Africa, with a single enigmatic species, Atypus snetsingeri Sarno, 1973, known only from a small area in southeastern Pennsylvania in eastern USA. A close relationship to European species could be assumed based on geographic proximity, but A. snetsingeri more closely resembled Asian species. This study was undertaken to learn more about the genetics of A. snetsingeri, its habitat requirements and natural history. Molecular markers (CO1 sequences) were compared to available data for other atypids and showed that A. snetsingeri is identical with A. karschi Dönitz, 1887 native to East Asia. Natural history parameters in Pennsylvania were also similar in every respect to A. karschi in Japan, therefore, we propose that the spider is an introduced species and the specific epithet snetsingeri is relegated to a junior synonym of A. karschi. Cytogenetic analysis showed an X0 sex chromosome system (42 chromosomes in females, 41 in males) and we also detected nucleolus organizing regions and heterochromatin, the latter for the first time in the Atypoidea. In Pennsylvania the spider is found in a variety of habitats, from forests to suburban shrubbery, where the above-ground webs are usually attached vertically to trees, shrubs, or walls, although other webs are oriented horizontally near the ground. Prey include millipedes, snails, woodlice, carabid beetles and earthworms. Atypus karschi is the first known case of an introduced purse-web spider. It is rarely noticed but well-established within its range in southeastern Pennsylvania.

BACKGROUND: Diabetes mellitus is unfavorably associated with cancer risk. The purpose of this multidisciplinary project was to evaluate a possible association of diabetes mellitus and other comorbidities and their treatment with progression of colorectal cancer. PATIENTS AND METHODS: We investigated the correlation between pathological characteristics and clinical course, including comorbidities in 1004 Czech patients diagnosed and surgically treated for colorectal adenocarcinoma (CRC) between 1999 and 2016. RESULTS: In our data set, CRC patients treated with metformin due to coexisting diabetes mellitus type 2 (T2DM) developed fewer distant metastases which clinically correlates with slower CRC progression. Survival in metformin subgroup was longer, particularly in men with CRC. Osteoporosis may be a negative factor of survival in CRC patients. CONCLUSIONS: Our findings also indicate that aging, higher tumor grade and TNM stage, coexistence of selected endocrine disorders, and metabolic abnormalities may change the tumor microenvironment and impact survival in colorectal cancer, although mechanism of these observations yet to be explained. Patients with diabetes mellitus type 2 treated with metformin may represent the altered microenvironment with specifically tuned metabolic molecular responses and with various epigenetic characteristics. More awareness and increased understanding of the mechanisms underlying the positive effect of metformin on patients' survival could offer insight into new treatment methods and permit more individualized treatment plans.

Beauverolides are hydrophobic cyclodepsipeptides that inhibit sterol O-acyltransferases and calmodulin, thereby reducing senile plaques in Alzheimer’s disease and preventing foam cell formation in atherosclerosis. Other sterol O-acyltransferase inhibitors suffer from low bioavailability, and evidence of the distribution of beauverolides in bodily fluids is lacking. We aimed to determine the optimal formulation of beauverolides for administration to experimental animals and to determine if the orally administered beauverolides could cross the gastrointestinal barrier and be secreted in urine. We found that beauverolides formed gels in aqueous solutions and we developed a formula for their peroral administration. Administration of the beauverolide gel pellets to experimental mice revealed that beauverolides cross the gastrointestinal barrier, circulate untouched in the blood, and are excreted in the urine. Using liquid chromatography-mass spectrometry analyses, we show that the administered beauverolides circulate in mouse blood and are excreted from the body 24 h after their administration.

Abstract Adenosine triphosphate (ATP) is a crucial substrate and energy source commonly used in enzyme reactions. However, we demonstrated that the addition of this acidic compound to enzyme assay buffers can serve as a source of unnoticed pH changes. Even relatively low concentrations of ATP (up to 5 mM) shifted pH of reaction mixtures to acidic values. For example, Tris buffer lost buffering capacity at pH 7.46 by adding ATP at a concentration higher than 2 mM. In addition to the buffering capacity, the pH shifts differed with respect to the buffer concentration. High ATP concentrations are commonly used in hexokinase assays. We demonstrated how the presence of ATP affects pH of widely used enzyme assay buffers and inversely affected K M of human hexokinase 2 and S 0.5 of human glucokinase. The pH optimum of human glucokinase was never reported before. We found that previously reported optimum of mammalian glucokinase was incorrect, affected by the ATP-induced pH shifts. The pH optimum of human glucokinase is at pH 8.5–8.7. Suggested is the full disclosure of reaction conditions, including the measurement of pH of the whole reaction mixtures instead of measuring pH prior to the addition of all the components.

Necropsy of two free-ranging common blackbirds (Turdus merula) found dead in central Italy revealed the presence of a high number of cyclocoelid flukes in the coelomatic cavity. Cyclocoelid flukes primarily infect avian respiratory system. Histologically, air sac walls were covered with a fibrinous exudate containing degenerate heterophils, many trematodes and some colonies of Gram-positive cocci. The superficial bronchi and parabronchi were markedly distended, and the adjacent pulmonary parenchyma was congested and collapsed. Trematodes, surrounded by a mild suppurative to pyogranulomatous inflammatory reaction, were also observed on the pericardial, intestinal, kidney and hepatic serosal surfaces. The death of the two examined birds was likely due to the high parasite load and associated severe lesions. At parasitological examination, flukes showed a tongue-shaped elongate body, tapered anteriorly and rounded posteriorly, of 2,088–2,314 μm in width and 8,268–11,830 μm in length. The mouth was slightly oval and sub-terminal, with a small oral sucker. The oval pharynx measured 250–309 μm, and the two caeca joined posteriorly. Two large (550–702 μm × 450–520 μm) globular testes were situated obliquely to each other, whereas an oval (250 × 300 μm in mean) or round (about 334 μm in diameter) intertesticular ovary was placed in a longitudinal straight line with the testes. The ootype was about 110 μm in diameter, while the brown-yellow eggs measured 131.5 × 73.9 μm in mean. The genital pore was post-pharyngeal, while the symmetrically arranged vitelline glands were not confluent posteriorly. Morphoflogical diagnosis led to the identification of Morishitium polonicum, a cyclocoelid fluke species that typically inhabits the air sacs of blackbirds. The morphological diagnosis was corroborated by molecular phylogenetic analysis of the mitochondrial (CO1, ND1) DNA loci. The present study provides the first report of pathological lesions and death caused by M. polonicum in birds. Keywords: Air sac trematodes, Cyclocoelidae, Death, Histopathological lesions, Morishitium polonicum, Turdus merula

Abstract Background Peritoneal larval cestodiasis induced by Mesocestoides Vaillant, 1863 (Cyclophyllidea: Mesocestoididae) is a common cause of severe infections in domestic dogs and cats, reported also from other mammals and less frequently from birds. However, there is a limited knowledge on the taxonomy of causative agents of this disease. Results In the present study, we investigated a massive, likely lethal, infection of a song thrush Turdus philomelos (Passeriformes: Turdidae) by Mesocestoides sp. tetrathyridia. We performed combined morphological and phylogenetic analysis of the tetrathyridia and compared them with the materials obtained previously from other birds and mammals. The metrical data fitted within the wide range reported by previous authors but confirmed the limited value of morphological data for species identification of tetrathyridia of Mesocestoides spp. The molecular analyses suggested that the isolates represented an unidentified Mesocestoides sp. that was previously repeatedly isolated and sequenced in larval and adult forms from domestic dogs and cats in Europe, the Middle East and North Africa. In contrast to the present study, which found encysted tetrathyridia, four of the five previous studies that identified the same species described infections by acephalic metacestodes only. Conclusions The tetrathyridia of the examined Mesocestoides sp. are described in the present study for the first time. However, the possible match with the species that were previously reported to infect birds remains uncertain. The phylogenetic analyses also suggested the rejection of two cases that were previously identified as Mesocestoides corti as they were likely caused by the same species as in the presently reported infection case. The newly provided DNA sequences should allow the assignment to species in the future, when adults of the genus Mesocestoides are more thoroughly sequenced.

Wasps of the genus Gasteruption are predator-inquilines of bees nesting in cavities in wood, stems, galls, and vertical soil surfaces. During studies of hymenopterans associated with reed galls caused by flies of the genus Lipara we recorded three species. We provide the evidence that a rare European species Gasteruption phragmiticola is a specialized predator-inquiline of an equally rare wetland bee Hylaeus pectoralis. Gasteruption nigrescens is a predator-inquiline of bees of the family Megachilidae, using the common bee Hoplitis leucomelana as the main host. Gasteruption assectator is a less specialized predator-inquiline of multiple groups of bees. The first two species, G. phragmiticola and G. nigrescens, are usually present at well-preserved reed beds associated with meadows rich in flowering plants while G. assectator does not prefer any specific habitat. Mature larvae of Gasteruption typically have a dorsoventrally flattened body and lateral lobes, small head with only slightly sclerotized mouthparts and tridentate mandibles. The larvae of these three species differ only slightly from one another, at most by the sclerotization of the mouthparts, colour, and position of setae on the head capsule and the shape of the mandible. As previous descriptions are insufficient, we provide first detailed descriptions with figures.

Tarantula urticating setae are modified setae located on the abdomen or pedipalps, which represent an effective defensive mechanism against vertebrate or invertebrate predators and intruders. They are also useful taxonomic tools as morphological characters facilitating the classification of New World theraphosid spiders. In the present study, the morphology of urticating setae was studied on 144 taxa of New World theraphosids, including ontogenetic stages in chosen species, except for species with urticating setae of type VII. The typology of urticating setae was revised, and types I, III and IV were redescribed. The urticating setae in spiders with type I setae, which were originally among type III or were considered setae of intermediate morphology between types I and III, are newly considered to be ontogenetic derivatives of type I and are described as subtypes. Setae of intermediate morphology between that of body setae and type II urticating setae that were found in Iridopelma hirsutum and Antillena rickwesti may provide another evidence that type II urticating setae evolved from body setae. It is supposed that the fusion of barbs with the shaft may lead to the morphology of type II setae. As the type II setae of Aviculariinae evolved independently to the UrS of Theraphosinae and both subfamilies represent two non-sister groups, this should explain the differences in the morphology of body setae in Aviculariinae and Theraphosinae. The terminology of "barbs" and "reversed barbs" was revised and redefined, newly emphasizing the real direction of barbs.

Abstract Background The cutaneous monostome trematode Collyriclum faba (Bremser in Schmalz, 1831) is a bird parasite with a hitherto unknown life cycle and highly focal occurrence across the Holarctic and Neotropic ecozones. Methods Representative specimens of benthic organisms were sampled at multiple sites and dates within the known foci of C. faba occurrence in Slovakia. A combined approach involving detailed morphological examination and sequencing of two independent DNA loci was used for their analysis. Results We elucidated the complete life cycle of C. faba, which we determined to include the aquatic gastropod mollusk Bythinella austriaca (Frauenfeld, 1857) as the first intermediate host, the mayflies of the family Heptageniidae, Ecdyonurus venosus (Fabricius, 1775) and Rhithrogena picteti Sowa, 1971 x iridina (Kolenati, 1839), as the second intermediate hosts, and birds (primarily but not exclusively passeriform birds) as the definitive hosts. Bythinella austriaca occurs focally in the springs of tributaries of the Danube in the Alpine-Carpathian region. The restricted distribution of B. austriaca explains the highly focal distribution of C. faba noticed previously in spite of the broad distribution of its second intermediate and definitive host species. Utilization of both larval and adult Ephemeroptera spp. as the second intermediate hosts explains the known spectrum of the definitive host species, with the highest prevalence in species feeding on larvae of Ephemeroptera, such as Cinclus cinclus (Linnaeus, 1758) and Motacilla cinerea Tunstall, 1771, or adults of Ephemeroptera, such as Sylvia atricapilla (Linnaeus, 1758) and Regulus regulus (Linnaeus, 1758). In this study, we also determine the prevalence and DNA sequences of other immature trematode specimens found in the examined benthic organisms (particularly the families Microphallidae, Troglotrematidae and Nanophyetidae and Euryhelmis zelleri Grabda-Kazubska, 1980, Heterophyidae), and describe cercariae of C. faba. Conclusions We determined the full life cycle of the Central European populations of C. faba. We speculate that other species of Bythinella and the closely related genus Amnicola may serve as first intermediate hosts in other parts of the distribution range of C. faba. Similarly, other Ephemeroptera of the family Heptageniidae may serve as the second intermediate hosts of C. faba in the Americas.

The ability of aculeate Hymenoptera to utilize wetlands is poorly understood, and descriptions of their nests and developmental stages are largely absent. Here we present results based on our survey of hymenopterans using galls induced by Lipara spp. flies on common reed Phragmites australis in the years 2015-2016. We studied 20,704 galls, of which 9,446 were longitudinally cut and the brood from them reared in the laboratory, while the remaining 11,258 galls reared in rearing bags also in laboratory conditions. We recorded eight species that were previously not known to nest in reed galls: cuckoo wasps Chrysis rutilans and Trichrysis pumilionis, solitary wasps Stenodynerus chevrieranus and Stenodynerus clypeopictus, and bees Pseudoanthidium tenellum, Stelis punctulatissima, Hylaeus communis and Hylaeus confusus. Forty five species of Hymenoptera: Aculeata are known to be associated with reed galls, of which 36 make their nests there, and the other are six parasitoids of the family Chrysididae and three cuckoo bees of the genus Stelis. Of these species, Pemphredon fabricii and in southern Europe also Heriades rubicola are very common in reed galls, followed by Hylaeus pectoralis and two species of the genus Trypoxylon. We also found new host-parasite associations: Chrysis angustula in nests of Pemphredon fabricii, Chrysis rutilans in nests of Stenodynerus clypeopictus, Trichrysis pumilionis in nests of Trypoxylon deceptorium, and Stelis breviuscula in nests of Heriades rubicola. We provide new descriptions of the nests of seven species nesting in reed galls and morphology of mature larvae of eight species nesting in reed galls and two parasitoids and one nest cleptoparasite. The larvae are usually very similar to those of related species but possess characteristics that make them easy to distinguish from related species. Our results show that common reeds are not only expansive and harmful, but very important for many insect species associated with habitats dominated by this plant species.

Among Accipitriformes sensu stricto, only a few species have been reported to form hybrid zones; these include the red kite Milvus milvus and black kite Milvus migrans migrans. M. milvus is endemic to the western Palearctic and has an estimated total population of 20-24,000 breeding pairs. The species was in decline until the 1970s due to persecution and has declined again since the 1990s due to ingestion of rodenticide-treated baits, illegal poisoning and changes in agricultural practices, particularly in its core range. Whereas F1 M. milvus × M. migr. migrans hybrid offspring have been found, F2 and F3 hybrids have only rarely been reported, with low nesting success rates of F1 hybrids and partial hybrid sterility likely playing a role. Here, we analyzed the mitochondrial (CO1 and CytB) and nuclear (Myc) DNA loci of 184 M. milvus, 124 M. migr. migrans and 3 F1 hybrid individuals collected across central Europe. In agreement with previous studies, we found low heterozygosity in M. milvus regardless of locus. We found that populations of both examined species were characterized by a high gene flow within populations, with all of the major haplotypes distributed across the entire examined area. Few haplotypes displayed statistically significant aggregation in one region over another. We did not find mitochondrial DNA of one species in individuals with the plumage of the other species, except in F1 hybrids, which agrees with Haldane´s Rule. It remains to be investigated by genomic methods whether occasional gene flow occurs through the paternal line, as the examined Myc gene displayed only marginal divergence between M. milvus and M. migr. migrans. The central European population of M. milvus is clearly subject to free intraspecific gene flow, which has direct implications when considering the origin of individuals in M. milvus re-introduction programs.

OBJECTIVE:AIDS-related mortality has changed dramatically with the onset of highly active antiretroviral therapy (HAART), which has even allowed compensated HIV-infected patients to withdraw from secondary therapy directed against opportunistic pathogens. However, in recently autopsied HIV-infected patients, we observed that associations with a broad spectrum of pathogens remain, although detailed analyses are lacking. Therefore, we focused on the possible frequency and spectrum shifts in pathogens associated with autopsied HIV-infected patients. DESIGN:We hypothesized that the pathogens frequency and spectrum changes found in HIV-infected patients examined postmortem did not recapitulate the changes found previously in HIV-infected patients examined antemortem in both the pre- and post-HAART eras. Because this is the first comprehensive study originating from Central and Eastern Europe, we also compared our data with those obtained in the West and Southwest Europe, USA and Latin America. METHODS:We performed autopsies on 124 HIV-infected patients who died from AIDS or other co-morbidities in the Czech Republic between 1985 and 2014. The pathological findings were retrieved from the full postmortem examinations and autopsy records. RESULTS:We collected a total of 502 host-pathogen records covering 82 pathogen species, a spectrum that did not change according to patients' therapy or since the onset of the epidemics, which can probably be explained by the fact that even recently deceased patients were usually decompensated (in 95% of the cases, the last available CD4+ cell count was falling below 200 cells*μl-1) regardless of the treatment they received. The newly identified pathogen taxa in HIV-infected patients included Acinetobacter calcoaceticus, Aerococcus viridans and Escherichia hermannii. We observed a very limited overlap in both the spectra and frequencies of the pathogen species found postmortem in HIV-infected patients in Europe, the USA and Latin America. CONCLUSIONS:The shifts documented previously in compensated HIV-infected patients examined antemortem in the post-HAART era are not recapitulated in mostly decompensated HIV-infected patients examined postmortem.

Bibliometric indicators increasingly affect careers, funding, and reputation of individuals, their institutions and journals themselves. In contrast to author self-citations, little is known about kinetics of journal self-citations. Here we hypothesized that they may show a generalizable pattern within particular research fields or across multiple fields. We thus analyzed self-cites to 60 journals from three research fields (multidisciplinary sciences, parasitology, and information science). We also hypothesized that the kinetics of journal self-citations and citations received from other journals of the same publisher may differ from foreign citations. We analyzed the journals published the American Association for the Advancement of Science, Nature Publishing Group, and Editura Academiei Române. We found that although the kinetics of journal self-cites is generally faster compared to foreign cites, it shows some field-specific characteristics. Particularly in information science journals, the initial increase in a share of journal self-citations during post-publication year 0 was completely absent. Self-promoting journal self-citations of top-tier journals have rather indirect but negligible direct effects on bibliometric indicators, affecting just the immediacy index and marginally increasing the impact factor itself as long as the affected journals are well established in their fields. In contrast, other forms of journal self-citations and citation stacking may severely affect the impact factor, or other citation-based indices. We identified here a network consisting of three Romanian physics journals Proceedings of the Romanian Academy, Series A, Romanian Journal of Physics, and Romanian Reports in Physics, which displayed low to moderate ratio of journal self-citations, but which multiplied recently their impact factors, and were mutually responsible for 55.9%, 64.7% and 63.3% of citations within the impact factor calculation window to the three journals, respectively. They did not receive nearly any network self-cites prior impact factor calculation window, and their network self-cites decreased sharply after the impact factor calculation window. Journal self-citations and citation stacking requires increased attention and elimination from citation indices.

Wetland species of aculeate Hymenoptera are poorly known, even though many of them may serve as diagnostic or flagship species in nature conservation. Here we examined 6,018 galls induced ≥1 year prior their collection by the chloropid flies Lipara spp. The galls were collected at 34 sites in Central Europe. We examined 1,389 nests (4,513 individuals) of nine species, part of which were parasitized by one dipteran and two chrysidid parasitoid species. We describe the nests of seven dominant species and larvae of four species (Pemphredon fabricii, Trypoxylon deceptorium, Hoplitis leucomelana and Hylaeus pectoralis) and two parasitoids (Trichrysis cyanea and Thyridanthrax fenestratus, both in nests of Pemphredon fabricii and Trypoxylon deceptorium). All the species, but H. pectoralis, preferred robust galls at very thin stalks (induced typically by Lipara lucens) over the narrow galls on thick stalks. The larvae of P. fabricii and T. deceptorium resembled strongly their sibling species (Pemphredon lethifer and Trypoxylon attenuatum sensu lato, respectively). The larvae of T. fenestratus showed features different from those previously described. By hatching set of another 10,583 galls induced by Lipara spp. ≥1 year prior their collection, we obtained 4,469 individuals of 14 nesting hymenopteran species, two cleptoparasites, three chrysidid and one dipteran parasitoid. Of these species, four new nesting species have been recorded for the first time in galls induced by Lipara spp.: Chelostoma campanularum, Heriades rubicola, Pseudoanthidium lituratum and Hylaeus incongruus. We also provide first records of their nest cleptoparasites Stelis breviuscula and Stelis ornatula, and the parasitoid Holopyga fastuosa generosa. Thyridanthrax fenestratus formed strong populations in nests of Pemphredon fabricii and Trypoxylon deceptorium, which are both newly recorded hosts for T. fenestratus. The descriptions provided here allow for the first time to identify the larvae of the most widespread central European aculeate hymenopteran reed gall specialists.

Pimecrolimus (Elidel, SDZ ASM 981) is an anti-inflammatory and immunomodulatory 33-epichloro-derivative of macrolactam ascomycin, with low potential for affecting systemic immune responses compared with other calcineurin inhibitors, cyclosporin A and tacrolimus. Despite numerous studies focused on the mechanism of pimecrolimus action on mast cells, only the single report has addressed pimecrolimus effects on other typical FcεRI-expressing cells, the basophils. Patients allergic to birch pollen (n = 20), hymenopteran venoms (n = 23) and 10 non-allergic volunteers were examined. Primary human basophils pre-treated or not with 0.5-50 μMol pimecrolimus were exposed to various concentrations of recombinant Bet v 1a allergen, bee or wasp venom extracts and anti-IgE for 20 min, and then examined for the expression of CD45, CD193, CD203c, CD63 and CD164 using flow cytometry. The externalization of basophil activation markers (CD63 and CD164) was equally inhibited through pimecrolimus in cells activated by recombinant pollen allergen, hymenopteran venom extracts and anti-IgE. Although the individual response rate was subject to strong variation, importantly, pre-treatment with pimecrolimus lowered the number of activated basophils in response to any of the stimuli in the basophils from all patients. The inhibition was concentration-dependent; approximately half of the basophils were inhibited in the presence of 2.5 mMol pimecrolimus. Pimecrolimus is a valuable new tool for the inhibition of hyper-reactive basophils in patients with pollen allergy and a history of anaphylactic reactions to bee or wasp venoms. Further research should address short-term use of pimecrolimus in vivo in a wide spectrum of allergic diseases.

BACKGROUND: Publication lag between manuscript submission and its final publication is considered as an important factor affecting the decision to submit, the timeliness of presented data, and the scientometric measures of the particular journal. Dual-format peer-reviewed journals (publishing both print and online editions of their content) adopted a broadly accepted strategy to shorten the publication lag: to publish the accepted manuscripts online ahead of their print editions, which may follow days, but also years later. Effects of this widespread habit on the immediacy index (average number of times an article is cited in the year it is published) calculation were never analyzed. METHODOLOGY/PRINCIPAL FINDINGS: Scopus database (which contains nearly up-to-date documents in press, but does not reveal citations by these documents until they are finalized) was searched for the journals with the highest total counts of articles in press, or highest counts of articles in press appearing online in 2010-2011. Number of citations received by the articles in press available online was found to be nearly equal to citations received within the year when the document was assigned to a journal issue. Thus, online publication of in press articles affects severely the calculation of immediacy index of their source titles, and disadvantages online-only and print-only journals when evaluating them according to the immediacy index and probably also according to the impact factor and similar measures. CONCLUSIONS/SIGNIFICANCE: Caution should be taken when evaluating dual-format journals supporting long publication lag. Further research should answer the question, on whether the immediacy index should be replaced by an indicator based on the date of first publication (online or in print, whichever comes first) to eliminate the problems analyzed in this report. Information value of immediacy index is further questioned by very high ratio of authors' self-citations among the citation window used for its calculation.