Phagocytosis is an important function of both insect haemocytes and mammalian blood cells. Linden bugs and cotton leaf worms have been suggested as new alternative models for ecological and drug toxicology but no data on their haemocyte physiology have been published. Our assays with particle ingestion of the NBT test were carried out on prohaemocytes, granulocytes, plasmatocytes and spherulocytes of adult linden bug and cotton leaf worm larvae. We found that phagocytic activity is on average 10% in the linden bug, and 50% in cotton leaf worm haemocytes: the phagocytic index is 3.5 in both species and nitroblue tetrazolium reduction is 0.5 in the linden bug and 3.2 in the cotton leaf worm. Phagocytic charactersitics of the prohaemocytes and granulocytes in the cotton leaf worm are closed to mammalian neutrophil physiology. Our data suggest that cotton leaf worm haemocytes may be a new potential alternative model for screening of phagocytosis.

• Klíčová slova
• Spodoptera littoralis, Pyrrhocoris apterus, NBT test,
• MeSH
• fagocytóza fyziologie   MeSH
• financování organizované MeSH
• hemocyty fyziologie   imunologie   MeSH
• Heteroptera fyziologie   imunologie   MeSH
• hmyz fyziologie   imunologie   MeSH
• krevní buňky fyziologie   imunologie   MeSH
• modely u zvířat MeSH
• Spodoptera fyziologie   imunologie   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH

The appearance of wings in insects, early in their evolution [1], has been one of the more critical innovations contributing to their extraordinary diversity. Despite the conspicuousness and importance of wings, the origin of these structures has been difficult to resolve and represented one of the "abominable mysteries" in evolutionary biology [2]. More than a century of debate has boiled the matter down to two competing alternatives-one of wings representing an extension of the thoracic notum, the other stating that they are appendicular derivations from the lateral body wall. Recently, a dual model has been supported by genomic and developmental data [3-6], representing an amalgamation of elements from both the notal and pleural hypotheses. Here, we reveal crucial information from the wing pad joints of Carboniferous palaeodictyopteran insect nymphs using classical and high-tech techniques. These nymphs had three pairs of wing pads that were medially articulated to the thorax but also broadly contiguous with the notum anteriorly and posteriorly (details unobservable in modern insects), supporting their overall origin from the thoracic notum as well as the expected medial, pleural series of axillary sclerites. Our study provides support for the formation of the insect wing from the thoracic notum as well as the already known pleural elements of the arthropodan leg. These results support the unique, dual model for insect wing origins and the convergent reduction of notal fusion in more derived clades, presumably due to wing rotation during development, and they help to bring resolution to this long-standing debate.

• MeSH
• biologická evoluce * MeSH
• hmyz anatomie a histologie   růst a vývoj   MeSH
• křídla zvířecí anatomie a histologie   MeSH
• nymfa anatomie a histologie   MeSH
• zkameněliny * MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

• MeSH
• bakteriální nemoci kůže MeSH
• dermatologické látky MeSH
• ekzematózní onemocnění kůže MeSH
• kožní nemoci farmakoterapie   MeSH
• papuloskvamózní nemoci kůže MeSH
• parazitární onemocnění kůže MeSH
• vezikulobulózní nemoci kůže MeSH
• virová kožní onemocnění MeSH

• Konspekt
• Lékařské vědy. Lékařství
• NLK Obory
• farmacie a farmakologie
• dermatovenerologie
• dermatovenerologie
• farmakoterapie
• farmakoterapie
• NLK Publikační typ
• publikace WHO

The CRISPR/Cas9 technique is widely used in experimentation with human cell lines as well as with other model systems, such as mice Mus musculus, zebrafish Danio reiro, and the fruit fly Drosophila melanogaster. However, publications describing the use of CRISPR/Cas9 for genome editing in non-model organisms, including non-model insects, are scarce. The introduction of this relatively new method presents many problems even for experienced researchers, especially with the lack of procedures to tackle issues concerning the efficiency of mutant generation. Here we present a protocol for efficient genome editing in the non-model insect species Pyrrhocoris apterus. We collected data from several independent trials that targeted several genes using the CRISPR/Cas9 system and determined that several crucial optimization steps led to a remarkably increased efficiency of mutant production. The main steps are as follows: the timing of embryo injection, the use of the heteroduplex mobility assay as a screening method, in vivo testing of sgRNA efficiency, and G0 germline mosaicism screening. The timing and the method of egg injections used here need to be optimized for other species, but other here-described optimization solutions can be applied immediately for genome editing in other insect species.

• Publikační typ
• časopisecké články MeSH

Equine insect bite hypersensitivity (IBH) is a seasonal IgE-mediated dermatosis caused by bites of insects of the genus Culicoides. A familial predisposition for the disease has been shown but, except for the MHC, the genes involved have not been identified so far. An immunogenomic analysis of IBH was performed in a model population of Old Kladruby horses, all living in the same environment. Clinical signs of IBH were used as phenotypic manifestation of IBH. Furthermore, total serum IgE levels were determined in the sera of these horses and used as an independent phenotypic marker for the immunogenetic analysis. Single nucleotide polymorphisms (SNPs) in candidate immunity-related genes were used for association analyses. Genotypes composed of two to five genes encoding interferon gamma -IFNG, transforming growth factor beta 1 -TGFB1, Janus kinase 2 -JAK2, thymic stromal lymphopoietin -TSLP, and involucrin -IVL were associated with IBH, indicating a role of the genes in the pathogenesis of IBH. These findings were supported by analysis of gene expression in skin biopsies of 15 affected and 15 unaffected horses. Two markers associated with IBH, IFNG and TGFB1, showed differences in mRNA expression in skin biopsies from IBH-affected and non-affected horses (p<0.05). Expression of the gene coding for the CD14 receptor molecule -CD14 was different in skin biopsies at p<0.06. When total IgE levels were treated as binary traits, genotypes of IGHE, ELA-DRA, and IL10/b were associated with this trait. When treated as a continuous trait, total IgE levels were associated with genes IGHE, FCER1A, IL4, IL4R, IL10, IL1RA, and JAK2. This first report on non-MHC genes associated with IBH in horses is thus supported by differences in expression of genes known to play a role in allergy and immunity.

• MeSH
• alergeny imunologie   MeSH
• atopická dermatitida genetika   imunologie   veterinární   MeSH
• Ceratopogonidae imunologie   MeSH
• cytokiny genetika   MeSH
• exprese genu MeSH
• hlavní histokompatibilní komplex MeSH
• imunoglobulin E krev   MeSH
• interferon gama genetika   MeSH
• Janus kinasa 2 genetika   MeSH
• jednonukleotidový polymorfismus MeSH
• koně MeSH
• kousnutí a bodnutí hmyzem genetika   imunologie   veterinární   MeSH
• messenger RNA genetika   MeSH
• nemoci koní genetika   imunologie   MeSH
• proteinové prekurzory genetika   MeSH
• slinné proteiny a peptidy imunologie   MeSH
• transformující růstový faktor beta1 genetika   MeSH
• zvířata MeSH
• Check Tag
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The objective of this study was to assess the genetic parameters of insect bite hypersensitivity (IBH) in the Old Grey Kladruber horse, an original Czech warmblood horse breed. Insect bite hypersensitivity is a recurrent allergic skin disease affecting horses worldwide. Its etiology is multifactorial. The defect is genetically controlled, and the starting impulse is a bite by midges of the spp. and less frequently spp. Knowledge about the associated genes is limited. Horses were kept by the National Stud in Kladruby (1,146 measurements) and by 10 private breeders (63 measurements). The horses were visually scored by the same inspector over a period of 13 yr from 1996 to 2009. A linear logistic model with a binary variable was used for subsequent statistical analysis. The fixed effects of the farm, year of evaluation, age of evaluation, and sex were significant, and the nonsignificant effect was the intensity of greying. The animals from the National Stud were more strongly affected by IBH than animals from private studs. The National Stud is located near the Labe River, at an altitude of 206 m; there are bottomland pastures in the vicinity, blind river arms, and numerous pools and marshes, and the climate is mild. These conditions are convenient for horse keeping but also for the propagation of . The stallions were less affected than the mares. The occurrence in years of age was influenced by selection; scoring began in the first year of age, and when the young animal was repeatedly positive, it was eliminated from breeding. The estimated h differed significantly from 0, and the direct h were 0.626 in the animal model with a maternal effect, 0.359 in the animal model without a maternal effect, and 0.363 in the animal model without correlation between direct and maternal effect. The maternal h was 0.305. The estimated repeatabilities were 0.636 with a maternal effect, 0.615 without a maternal effect, and 0.623 without correlation. The h of IBH was found to be medium to high, so there is a possibility to reduce the prevalence of IBH by selection.

• MeSH
• alergie genetika   veterinární   MeSH
• genetická predispozice k nemoci * MeSH
• koně MeSH
• kousnutí a bodnutí hmyzem imunologie   veterinární   MeSH
• lineární modely MeSH
• modely genetické MeSH
• nemoci koní epidemiologie   genetika   imunologie   MeSH
• prevalence MeSH
• sexuální faktory MeSH
• stárnutí MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika MeSH

• MeSH
• algoritmy MeSH
• elektroforéza kapilární metody   MeSH
• hmyzí proteiny chemie   MeSH
• koncentrace vodíkových iontů MeSH
• oligopeptidy izolace a purifikace   MeSH
• teoretické modely MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH

A new screening test of a haemolytic potential is presented on Spodoptera larvae in vivo. The assay was evaluated using phenylhydrazine, the well-known xenobiotic which induces haemolysis in mammals. Haemocyte disintegration causes changes in total haemocyte counts and differentials. It is shown that only qualitative evaluation of panoptically stained haemolymph smears is sufficient, fast and cheap to detect haemolytic potential. The presented assay also prevents unneeded, non-ethical use of vertebrates.

• MeSH
• alternativy testů na zvířatech MeSH
• fenylhydraziny toxicita   MeSH
• hemocyty cytologie   účinky léků   MeSH
• hemolýza MeSH
• larva cytologie   MeSH
• modely u zvířat MeSH
• počet buněk MeSH
• Spodoptera cytologie   MeSH
• testy toxicity metody   MeSH
• xenobiotika toxicita   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• práce podpořená grantem MeSH

Two values of thermal requirements, the lower developmental threshold (LDT), that is, the temperature at which development ceases, and the sum of effective temperatures, that is, day degrees above the LDT control the development of ectotherms and are used in phenology models to predict time at which the development of individual stages of a species will be completed. To assist in the rapid development of phenology models, we merged a previously published database of thermal requirements for insects, gathered by online search in CAB Abstracts, with independently collected data for insects and mites from original studies. The merged database comprises developmental times at various constant temperatures on 1,054 insect and mite species, many of them in several populations, mostly pests and their natural enemies, from all over the world. We show that closely related species share similar thermal requirements and therefore, for a species with unknown thermal requirements, the value of LDT and sum of effective temperatures of its most related species from the database can be used.

• MeSH
• biologické modely MeSH
• druhová specificita MeSH
• hmyz růst a vývoj   MeSH
• roztoči růst a vývoj   MeSH
• teplota MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Stream ecosystems face ever-increasing pressures by the presence of emergent contaminants, such as, personal care products. N, N-diethyl-3-methylbenzamide (DEET) is a synthetic insect repellent that is being found in surface waters environments in concentrations up to 33.4 μg/L. Information concerning DEET's toxicity in the aquatic environment is still limited and focused only on its acute effects on model species. Our main objective was to assess the effects of DEET exposure to a caddisfly non-target species using sub-lethal endpoints. For that, we chose Sericostoma vittatum, an important shredder in Portuguese freshwaters that has been already used in different ecotoxicological assays. Besides acute tests, S. vittatum were exposed during 6 days to a gradient of DEET concentrations (8, 18 and 40.5 mg/L) to assess effects on feeding behaviour and biochemical responses, such as, lipid peroxidation levels (LPO), catalase and acetylcholinesterase (AChE) activities, and also assess effects on energy reserves and consumption. Acute tests revealed a 48 h-LC50 of 80.12 mg/L and DEET exposure caused feeding inhibition with a LOEC of 36.80 mg/L. Concerning the biochemical responses, DEET caused no effects in LPO nor on catalase activity. A non-significant decrease in AChE activity was observed. Regarding energetic reserves, exposure to DEET caused a significant reduction in S. vittatum carbohydrates levels. These results add important information for the risk assessment of insect repellents in the aquatic environment and suggest that reported environmental concentrations of DEET are not toxic to non-target freshwater insects.

• MeSH
• chemické látky znečišťující vodu toxicita   MeSH
• diethyltoluamid toxicita   MeSH
• hmyz účinky léků   fyziologie   MeSH
• insekticidy toxicita   MeSH
• katalasa MeSH
• repelenty proti hmyzu toxicita   MeSH
• sladká voda MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH