Paenibacillus larvae and Melissococcus plutonius represent the most threatening bacterial diseases of honeybee (Apis mellifera)-American and European foulbrood, respectively. For efficient control of those diseases, rapid and accurate detection of the pathogens is crucial. Therefore, we developed a novel multiplex PCR method simultaneously detecting both pathogens. To design and optimize multiplex PCR reaction, four strains of P. larvae representing four ERIC genotypes I-IV (strain DSM 7030-ERIC I, DSM 25430-ERIC II, LMG 16252-ERIC III, DSM 3615-ERIC IV) were selected. Those strains were fully sequenced using long-read sequencing (Sequel I, Pacific Biosciences). For P. larvae, the multicopy insertion sequence IS256 identified in all genotypes of P. larvae was selected to provide high sensitivity. M. plutonius was detected by plasmid pMP1 sequence and the virulence verified by following detection of ETX/MTX2 toxin responsible for pore formation in the cell membrane. As an internal control, a gene encoding for major royal jelly protein 1 specific for honeybees was selected. The method was validated on 36 clinical specimens collected from the colonies suffering from American and European foulbrood in the Czech Republic. Based on the results, sensitivity of PCR was calculated to 93.75% and specificity to 100% for P. larvae diagnosed from hive debris and 100% sensitivity and specificity for honeybee workers and larval scales as well as for diseased brood infected by M. plutonius.

• Klíčová slova
• Apis mellifera, Melissococcus plutonius, Paenibacillus larvae, American foulbrood, European foulbrood, Multiplex PCR,
• MeSH
• Enterococcaceae * MeSH
• larva mikrobiologie   MeSH
• multiplexová polymerázová řetězová reakce metody   MeSH
• Paenibacillus larvae * genetika   MeSH
• Paenibacillus * genetika   MeSH
• plazmidy genetika   MeSH
• transpozibilní elementy DNA MeSH
• včely genetika   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• transpozibilní elementy DNA MeSH

The authors have been studying the bacterial diseases of the brood for a fairly long time. American and European foul brood has been studied with the highest attention. Cultures of Bacillus larvae (White, 1906) were examined both in freshly isolated strains and in collection strains of this micro-organism. In cases of foul brood, the pathological material was found to contain not only the typical rods of B. larvae but also immobile spiral forms which are usually referred to in literature as fragments or developmental forms of B. larvae. These spiral forms were found to constitute spindle-shaped formations in the culture of B. larvae; the multiplication of these spindles depends on the presence of the rods of B. larvae and their development and reproduction can be observed on wet gelatine agar in a Petri dish turned upside down under a normal microscope (10 X 10 magnification). In the combined liquid medium, used in the experiments, these formations disintegrate into immobile spirals; if re-cultivated on a solid medium they re-assume their spindle shape with transverse meridian arrangement (in different amounts). Staining for proving the presence of nucleic acids does not eliminate the possibility of these formations being separate micro-organisms which cannot be stained by current staining methods but can be represented by the contrast method according to Burri, or by silvering according to Klein. The authors succeeded to separate these micro-organisms, but without the rods of B. larvae the colonies of these formations are feeble.

• MeSH
• Bacillus cytologie   MeSH
• bakteriální infekce veterinární   MeSH
• larva mikrobiologie   MeSH
• včely * embryologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

American foulbrood (AFB) is a devastating disease of honey bees. There remains a gap in the understanding of the interactions between the causative agent and host, so we used shotgun proteomics to gain new insights. Nano-LC-MS/MS analysis preceded visual description and Paenibacillus larvae identification in the same individual sample. A further critical part of our methodology was that larvae before capping were used as the model stage. The identification of the virulence factors SplA, PlCBP49, enolase, and DnaK in all P. larvae-positive samples was consistent with previous studies. Furthermore, the results were consistent with the array of virulence factors identified in an in vitro study of P. larvae exoprotein fractions. Although an S-layer protein and a putative bacteriocin were highlighted as important, the microbial collagenase ColA and InhA were not found in our samples. The most important virulence factor identified was isoform of neutral metalloproteinase (UniProt: V9WB82), a major protein marker responsible for the shift in the PCA biplot. This protein is associated with larval decay and together with other virulence factors (bacteriocin) can play a key role in protection against secondary invaders. Overall, this study provides new knowledge on host-pathogen interactions and a new methodical approach to study the disease.

• Klíčová slova
• Apis mellifera, diagnostics with mass spectrometry, host-pathogen interaction, label-free proteomics, neutral metalloproteinase isoforms,
• MeSH
• bakteriociny * metabolismus   MeSH
• faktory virulence metabolismus   MeSH
• larva MeSH
• Paenibacillus larvae * metabolismus   MeSH
• Paenibacillus * metabolismus   MeSH
• proteomika MeSH
• tandemová hmotnostní spektrometrie MeSH
• včely MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Spojené státy americké MeSH
• Názvy látek
• bakteriociny * MeSH
• faktory virulence MeSH

Up to date we do not have much information about predation on seeds by larvae of ground beetles. One of the reasons why such knowledge is important is that granivorous larvae contribute to predation of weed seeds. In this study, the food requirements of larvae of autumn breeding carabid species Amara eurynota (Panzer) were investigated in the laboratory and a hypothesis, that they are granivorous was tested. Insect diet (Tenebrio molitor larvae), three seed diets (seeds of Artemisia vulgaris, Tripleurospermum inodorum or Urtica dioica or a mixed diet (T. molitor + A. uulgaris) were used as food. For larvae of A. eurynota, seeds are essential for successful completion of development, because all those fed pure insect diet died before pupation. However, differences in suitability were observed between pure seed diets. Larvae fed seeds of A. vulgaris had the lowest mortality and fastest development of the seed diets. Those fed seeds of T. inodorum had also low mortality, but the development was prolonged in the third instar. In contrast, development of larvae reared on seeds of U. dioica was slowest of the tested diets and could not be completed, as all individuals died before pupation. When insects were included to seed diet of A. vulgaris (mixed diet), the duration of development shortened, but mortality remained the same when compared to seed diet of A. vulgaris. According to the results it was concluded that larvae of A. eurynota are granivorous. A mixed diet and seed diets of A. uulgaris and T. inodorum were suitable and insect diet and seeds of U. dioica were unsuitable diets in this experiment.

• MeSH
• brouci růst a vývoj   MeSH
• larva MeSH
• predátorské chování MeSH
• semena rostlinná * MeSH
• Tenebrio růst a vývoj   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Paenibacillus larvae is the causative agent of American foulbrood (AFB), a devastating disease of honeybee larvae. In the Czech Republic, two large infested regions were recognised. This study aimed to analyse P. larvae strains occurring in the Czech Republic in the years 2016-2017 and to characterise the genetic structure of their population with the use of Enterobacterial Repetitive Intergenic Consensus genotyping (ERIC), multilocus sequence typing (MLST) and whole genome sequence (WGS) analysis. The results were complemented by the analysis of isolates collected in the year 2018 in areas of Slovakia located near the Czechia-Slovakia border. ERIC genotyping revealed that 78.9% of tested isolates belonged to the ERIC II genotype and 21.1% to ERIC I genotype. MLST showed six sequence types with ST10 and ST11 being the most frequent among isolates. Within six isolates we found discrepancies in correlations between MLST and ERIC genotypes. The use of MLST and WGS analysis of isolates revealed that each of the large infested geographic regions had its own dominating P. larvae strains. We assume that these strains represented primary sources of infection in the affected areas. In addition, the sporadic presence of strains identified by core genome analysis as genetically related was unveiled in geographically distant regions suggesting possible human-mediated transmission of AFB.

• Klíčová slova
• AFB spread, American foulbrood, Epidemiology, Genotyping, Honeybee,
• MeSH
• genomika MeSH
• genotyp MeSH
• larva genetika   mikrobiologie   MeSH
• lidé MeSH
• multilokusová sekvenční typizace veterinární   MeSH
• Paenibacillus larvae * genetika   MeSH
• včely MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika epidemiologie   MeSH
• Slovenská republika epidemiologie   MeSH
• Spojené státy americké MeSH

Paenibacillus larvae is the causative agent of American foulbrood (AFB), the most serious bacterial disease affecting developing honeybee larvae and pupas. In this study, a library of 24 (thio)glycosides, glycosyl sulfones, 6-O-esters, and ethers derived from d-mannose, d-glucose, and d-galactose having C10 or C12 alkyl chain were evaluated for their antibacterial efficacy against two P. larvae strains. The efficacy of the tested compounds determined as minimal inhibitory concentrations (MICs) varied greatly. Generally, dodecyl derivatives were found to be more potent than their decylated analogs. Thioglycosides were more efficient than glycosides and sulfones. The activity of the 6-O-ether derivatives was higher than that of their ester counterparts. Seven derivatives with dodecyl chain linked (thio)glycosidically or etherically at C-6 showed high efficacy against both P. larvae strains (MICs ranged from 12.5 μM to 50 μM). Their efficacies were similar or much higher than those of selected reference compounds known to be active against P. larvae-lauric acid, monolaurin, and honeybee larval food components, 10-hydroxy-2-decenoic acid, and sebacic acid (MICs ranged from 25 μM to 6400 μM). The high efficacies of these seven derivatives suggest that they could increase the anti-P. larvae activity of larval food and improve the resistance of larvae to AFB disease through their application to honeybee colonies.

• Klíčová slova
• 10-HDA, American foulbrood, antibacterial activity, carbohydrate (thio)ethers, carbohydrate esters, fatty acid, lauric acid, monolaurin, royal jelly,
• MeSH
• antibakteriální látky farmakologie   MeSH
• estery farmakologie   MeSH
• ethery farmakologie   MeSH
• glykosidy farmakologie   MeSH
• larva MeSH
• Paenibacillus larvae * MeSH
• Paenibacillus * MeSH
• sacharidy farmakologie   MeSH
• sulfidy farmakologie   MeSH
• sulfony farmakologie   MeSH
• včely MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Spojené státy americké MeSH
• Názvy látek
• antibakteriální látky MeSH
• estery MeSH
• ethery MeSH
• glykosidy MeSH
• sacharidy MeSH
• sulfidy MeSH
• sulfony MeSH

Chironomids (non-biting midges) inhabit almost every wet or semi-wet continental environment on Earth with probably 10,000 different species. Species occurrence and composition are undoubtedly limited by environmental harshness and food availability being reflected in their energy stores. Most animals store energy as glycogen and lipid. They enable the animals to survive adverse situations and to continue growth, development, and reproduction. This general statement is also true for insects and also particularly true for chironomid larvae. The rationale behind this research was, that probably any stress, any environmental burden, and any harmful influence increases the energy requirement of individual larvae depleting energy stores. We developed new methods to measure the glycogen and lipid content in small tissue samples. Here we show how to apply these methods to single chironomid larvae to demonstrate their energy stores. We compared different locations of the high Alpine rivers along harshness gradient densely populated and dominated by chironomid larvae. All samples show very low energy stores without any major differences. We found glycogen concentrations below 0.01% of dry weight (DW) and lipid concentrations below 5% of DW irrespective of the specific sampling point. These values are among the lowest ever observed in chironomid larvae. We demonstrate that individuals living in extreme environment are stressed leading to reduced energy stores in their bodies. This appears to be a general feature of high altitude regions. Our results provide new insights and a better understanding of population and ecological dynamics in harsh mountainous areas, also in view of a changing climate.

• Klíčová slova
• Alpine streams, Chironomidae - Diamesa larvae, Energy storage, Glycogen, Lipids, Physiological indicator,
• MeSH
• Chironomidae * MeSH
• larva MeSH
• lipidy MeSH
• řeky MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Rakousko MeSH
• Názvy látek
• lipidy MeSH

Trilobites represent a model for 'evo-devo' studies in fossil euarthropods, owing to a rare developmental trait: the biomineralization of the dorsal exoskeleton soon after hatching. Many fossilized trilobite ontogenies thus feature early stages - the protaspides - characterized by non-articulated, calcified dorsal exoskeletons. The recent discovery of a protaspid-like fossil occurring with aglaspidid remains in Middle Ordovician strata of Siberia has been interpreted as evidence for the presence of protaspides in these distant relatives of trilobites. In this contribution, we demonstrate that this Siberian protaspis actually belongs to the asaphid trilobite Isotelus (or a related taxon), a genus likely present in the same bed. We conclude that protaspid larvae still represent a developmental trait unique to Trilobita.

• Klíčová slova
• Aglaspidida, Euarthropoda, Larval ecology, Ordovician, Protaspis, Trilobita,
• MeSH
• členovci * MeSH
• larva MeSH
• zkameněliny MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

The metastrongyloid nematode Angiostrongylus cantonensis causes eosinophilic meningitis in a variety of homeothermic hosts including humans. Third-stage infectious larvae develop in gastropods as intermediate hosts. Humans are usually infected by intentional or incidental ingestion of an infected mollusk or paratenic host (poikilothermic vertebrates and invertebrates). The infection may also hypothetically occur through ingestion of food or water contaminated by third-stage larvae spontaneously released from gastropods. Larvae are thought to be released in greater numbers from the intermediate host exposed to stress. This study aimed to compare larval release from stressed with unstressed gastropods. Experimentally infected Limax maximus and Lissachatina fulica were exposed to a stress stimulus (shaking on an orbital shaker). The mucus was collected before and after the stress and examined microscopically and by qPCR for the presence of A. cantonensis larvae and their DNA. In the case of L. maximus, no larvae were detected microscopically in the mucus, but qPCR analysis confirmed the presence of A. cantonensis DNA in all experimental replicates, without clear differences between stressed and non-stressed individuals. In contrast, individual larvae of A. cantonensis were found in mucus from Li. fulica after stress exposure, which also reflects an increased number of DNA-positive mucus samples after stress. Stress stimuli of intensity similar to the transport or handling of mollusks can stimulate the release of larvae from highly infected intermediate hosts. However, these larvae are released in small numbers. The exact number of larvae required to trigger neuroangiostrongyliasis is unknown. Therefore, caution is essential when interacting with potential intermediate hosts in regions where A. cantonensis is endemic.

• Klíčová slova
• Angiostrongylus cantonensis, Gastropods, Larvae release, Stress stimuli,
• MeSH
• Angiostrongylus cantonensis * fyziologie   MeSH
• fyziologický stres * MeSH
• hlen MeSH
• infekce hlísticemi řádu Strongylida parazitologie   MeSH
• kvantitativní polymerázová řetězová reakce MeSH
• larva * fyziologie   MeSH
• plži parazitologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Larvae (metacestodes) of gryporhynchid tapeworms (Cestoda: Cyclophyllidea) are reported for the first time from the liver of tilapia hybrids (Oreochromis aureus × O. niloticus) reared in earth ponds in northeastern Israel (along the Jordan River). This is the first record of Amirthalingamia macracantha (Joyeux & Baer, 1935), a parasite of cormorants (Phalacrocoracidae), outside Africa and outside the tropics. Larvae found in the liver of tilapias (Cichlidae) were identified to species level because they possessed 20 massive rostellar hooks of 3 types, with the 4 largest hooks measuring almost 500 µm. Molecular data confirmed species identification. The possible route of introduction to Israel of this African parasite, which is large (length up to 2 cm) and potentially pathogenic for cultivated tilapias, is discussed.

• Klíčová slova
• Cestode larvae, Invasive parasite, Israel, Tilapia hybrids,
• MeSH
• Cestoda * genetika   MeSH
• larva MeSH
• Tilapia * MeSH
• vodní hospodářství MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Izrael MeSH