The class Mollicutes comprises microorganisms that lack a cell wall, highly dependent on their host to survive. Within Mollicutes, the genus Spiroplasma comprises motile helical microorganisms associated with various insects and other arthropods. This study aimed to detect and characterize Mollicutes microorganisms in ticks of different species of veterinary importance, using molecular techniques. These ticks were collected from dogs, cats, cattle, and horses from Rio de Janeiro's metropolitan regions. They were morphologically classified and pooled according to their species for subsequent DNA extraction. These samples were tested by PCR using class Mollicutes-specific primers (16S rRNA) and positive amplicons were sequenced. The obtained DNA sequences were compared with other Mollicutes sequences deposited in GenBank. We found that four out of 745 (0.54%) of the tick pools were positive for members of the class Mollicutes, identified as Spiroplasma spp.; of the positive pools, one comprised Amblyomma sculptum adults and three comprised Dermacentor nitens nymphs. The present study describes Spiroplasma spp. in ticks in Brazil for the first time. Nevertheless, due to few reports on these microorganisms, further studies on epidemiology, virulence, and pathogenicity are needed.

• MeSH
• klíšťata * MeSH
• koně MeSH
• nymfa MeSH
• RNA ribozomální 16S genetika   MeSH
• Spiroplasma * genetika   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Brazílie MeSH

Spiroplasma eriocheiris caused massive mortality of Chinese mitten crab Eriocheir sinensis but little is known about the molecular characteristics of this microorganism. We described here the identification of a spiralin-like protein (SLP31) from S. eriocheiris and expression in Escherichia coli. Analysis of the nucleotide sequence revealed that the clone has an open reading frame of 837 bp encoding a protein of 279 amino acids. Theoretical isoelectric point and molar mass for SLP31 are 7.72 and 31 kDa, respectively. The similarity of SLP31 deduced amino acid sequence shared with the spiralin from other species indicated that the gene may be a member of spiralin family. The TGA codon in Spiroplasma serves not as a stop signal but as a code for the amino acid tryptophan. After cloning the SLP31, the gene was site-mutated from TGA to TGG and transcribed in E. coli to full expression of SLP31. The purified recombinant protein was used to determine the immune reactivity by Western blotting which suggests that SLP31 could be a good antigen for immunodiagnostic of tremor disease in E. sinensis.

• MeSH
• antigeny bakteriální MeSH
• bakteriologické techniky metody   MeSH
• DNA bakterií genetika   chemie   MeSH
• Escherichia coli genetika   MeSH
• financování organizované MeSH
• fylogeneze MeSH
• imunoanalýza metody   MeSH
• izoelektrický bod MeSH
• klonování DNA MeSH
• krabi mikrobiologie   MeSH
• molekulární sekvence - údaje MeSH
• molekulová hmotnost MeSH
• otevřené čtecí rámce MeSH
• proteiny vnější bakteriální membrány MeSH
• sekvence aminokyselin MeSH
• sekvence nukleotidů MeSH
• sekvenční analýza DNA MeSH
• sekvenční homologie MeSH
• shluková analýza MeSH
• Spiroplasma genetika   imunologie   izolace a purifikace   MeSH
• western blotting MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Geografické názvy
• Čína MeSH

Bacteria associated with the tick Ixodes ricinus were assessed in specimens unattached or attached to the skin of cats, dogs and humans, collected in the Czech Republic. The bacteria were detected by PCR in 97 of 142 pooled samples including 204 ticks, i.e. 1-7 ticks per sample, collected at the same time from one host. A fragment of the bacterial 16S rRNA gene was amplified, cloned and sequenced from 32 randomly selected samples. The most frequent sequences were those related to Candidatus Midichloria midichlori (71% of cloned sequences), followed by Diplorickettsia (13%), Spiroplasma (3%), Rickettsia (3%), Pasteurella (3%), Morganella (3%), Pseudomonas (2%), Bacillus (1%), Methylobacterium (1%) and Phyllobacterium (1%). The phylogenetic analysis of Spiroplasma 16S rRNA gene sequences showed two groups related to Spiroplasma eriocheiris and Spiroplasma melliferum, respectively. Using group-specific primers, the following potentially pathogenic bacteria were detected: Borellia (in 20% of the 142 samples), Rickettsia (12%), Spiroplasma (5%), Diplorickettsia (5%) and Anaplasma (2%). In total, 68% of I. ricinus samples (97/142) contained detectable bacteria and 13% contained two or more putative pathogenic groups. The prevalence of tick-borne bacteria was similar to the observations in other European countries.

• MeSH
• Bacteria klasifikace   genetika   MeSH
• DNA bakterií genetika   MeSH
• fylogeneze MeSH
• klíště růst a vývoj   mikrobiologie   MeSH
• kočky mikrobiologie   parazitologie   MeSH
• lidé MeSH
• molekulární sekvence - údaje MeSH
• nymfa mikrobiologie   MeSH
• polymerázová řetězová reakce MeSH
• psi mikrobiologie   parazitologie   MeSH
• RNA ribozomální 16S genetika   MeSH
• Spiroplasma klasifikace   genetika   MeSH
• velkoměsta MeSH
• zvířata MeSH
• Check Tag
• kočky mikrobiologie   parazitologie   MeSH
• lidé MeSH
• mužské pohlaví MeSH
• psi mikrobiologie   parazitologie   MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH
• velkoměsta MeSH

The ectoparasitic mite Varroa destructor is a major pest of the honeybee Apis mellifera. In a previous study, bacteria were found in the guts of mites collected from winter beehive debris and were identified using Sanger sequencing of their 16S rRNA genes. In this study, community comparison and diversity analyses were performed to examine the microbiota of honeybees and mites at the population level. The microbiota of the mites and honeybees in 26 colonies in seven apiaries in Czechia was studied. Between 10 and 50 Varroa females were collected from the bottom board, and 10 worker bees were removed from the peripheral comb of the same beehive. Both bees and mites were surface sterilized. Analysis of the 16S rRNA gene libraries revealed significant differences in the Varroa and honeybee microbiota. The Varroa microbiota was less diverse than was the honeybee microbiota, and the relative abundances of bacterial taxa in the mite and bee microbiota differed. The Varroa mites, but not the honeybees, were found to be inhabited by Diplorickettsia. The relative abundance of Arsenophonus, Morganella, Spiroplasma, Enterococcus, and Pseudomonas was higher in Varroa than in honeybees, and the Diplorickettsia symbiont detected in this study is specific to Varroa mites. The results demonstrated that there are shared bacteria between Varroa and honeybee populations but that these bacteria occur in different relative proportions in the honeybee and mite bacteriomes. These results support the suggestion of bacterial transfer via mites, although only some of the transferred bacteria may be harmful.

• MeSH
• biodiverzita MeSH
• DNA bakterií genetika   MeSH
• mikrobiota * MeSH
• RNA ribozomální 16S genetika   MeSH
• roční období MeSH
• sekvenční analýza DNA MeSH
• Spiroplasma klasifikace   izolace a purifikace   MeSH
• symbióza MeSH
• Varroidae mikrobiologie   MeSH
• včely mikrobiologie   parazitologie   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• srovnávací studie MeSH

Celkem jsme v roce 2007 vyšetřili 1048 jedinců klíštěte Ixodes ricinus z toho 935 ze sběrů vlajkováním v pražských parcích a 114 jedinců ze zákusu na člověku. V roce 2008 jsme vyšetřili 364 jedinců z toho 277 ze sběru a 86 sejmutých z lidí. Promořenost klíšťat původcem lymeské borreliózy v roce 2007 byla 9,7 %, zjištěná v temném poli a v 13 % byla prokázána DNA pomocí PCR. V roce 2008 byla promořenost klíšťat vyšší, 14 % v temném poli a 20 % pomocí PCR. Klíšťata sejmutá po zákusu na lidech byla infikována v 7 %. Přítomnost Anaplasma phagocytophilum, Bartonella sp. a Rickettsia sp byla prokázána pomocí polymerázové řetězové reakce a kontrolována sekvencí geonomu u 11,3 - 11,8 % klíšťat. Spiroplasma rodu Babesia byla zjištěna v 7,4 % u samic I. ricinus. Klíšťata sejmutá z lidí byla infikovaná borrelií v 6,5 % a anaplasmou v 1,5 %. Pravidelnými sběry v jednotlivých měsících r. 2007 a 2008 na jedné lokalitě v Praze 10 jsme ověřili změny výskytu ruzných stádií klíštěte a jejich měnící se nákazu ruznými agens v závislosti na teplotě a vývoji. K růstu bakterií Borrelia a Anaplasma sp. ve střevě klíštěte je zapotřebí určité doby s vyšší teplotou. Nejmenší počet infikovaných klíšťat byl v srpnu 2007 a 2008, kdy se z velké většiny vyskytovaly pouze larvy.

In 2007, 1048 Ixodes ricinus ticks were investigated: 935 of these were collected by flagging in Prague parks and 113 were ticks attached to the human skin. In 2008, 364 ticks were investigated, i.e. 277 and 87 ticks collected by flagging and from humans, respectively. In 2007, the causative agent of lyme borreliosis was detected in 9.7% of ticks in dark field and in 13% of ticks in DNA by PCR. In 2008, higher positivity rates were found, i.e. 14% and 20%, respectively. Seven percent of ticks obtained from humans were infected. Anaplasma phagocytophilum, Bartonella sp. and Rickettsia sp. were detected by PCR and sequence analysis in 11.3% - 11.8% of ticks. Spiroplasma of the genus Babesia was detected in 7.4% of I. ricinus females. The ticks collected from humans were infected by Borrelia in 6.5% and by Anaplasma in 1.5%. Regular monthly flagging was performed in one locality in Prague 10 in 2007 and 2008 to obtain data on the incidence of different stages of ticks and rates of infection by different agents depending on temperature and season. To grow in the tick intestine, Borrelia and Anaplasma need higher temperature for a certain period of time. The lowest numbers of infected ticks were found in August 2007 and 2008 when mostly tick larvae were collected.

• MeSH
• Anaplasma phagocytophilum genetika   izolace a purifikace   MeSH
• Babesia genetika   izolace a purifikace   MeSH
• Bartonella genetika   izolace a purifikace   MeSH
• Borrelia burgdorferi genetika   izolace a purifikace   MeSH
• infekce přenášené vektorem MeSH
• klíště mikrobiologie   MeSH
• lymeská nemoc epidemiologie   přenos   MeSH
• polymerázová řetězová reakce využití   MeSH
• Publikační typ
• grafy a diagramy MeSH

AIMS: The winter beehive debris containing bodies of honeybee parasitic mite Varroa destructor is used for veterinary diagnostics. The Varroa sucking honeybee haemolymph serves as a reservoir of pathogens including bacteria. Worker bees can pick up pathogens from the debris during cleaning activities and spread the infection to healthy bees within the colony. The aim of this study was to detect entomopathogenic bacteria in the Varroa collected from the winter beehive debris. METHODS AND RESULTS: Culture-independent approach was used to analyse the mite-associated bacterial community. Total DNA was extracted from the samples of 10 Varroa female individuals sampled from 27 different sites in Czechia. The 16S rRNA gene was amplified using universal bacterial primers, cloned and sequenced, resulting in a set of 596 sequences representing 29 operational taxonomic units (OTU97). To confirm the presence of bacteria in Varroa, histological sections of the mites were observed. Undetermined bacteria were observed in the mite gut and fat tissue. CONCLUSION: Morganella sp. was the most frequently detected taxon, followed by Enterococcus sp., Pseudomonas sp., Rahnella sp., Erwinia sp., and Arsenophonus sp. The honeybee putative pathogen Spiroplasma sp. was detected at one site and Bartonella-like bacteria were found at four sites. PCR-based analysis using genus-specific primers enabled detection of the following taxa: Enterococcus, Bartonella-like bacteria, Arsenophonus and Spiroplasma. SIGNIFICANCE AND IMPACT OF THE STUDY: We found potentially pathogenic (Spiroplasma) and parasitic bacteria (Arsenophonus) in mites from winter beehive debris. The mites can be reservoirs of the pathogenic bacteria in the apicultures.

• MeSH
• Bacteria klasifikace   genetika   izolace a purifikace   MeSH
• fylogeneze MeSH
• molekulární sekvence - údaje MeSH
• RNA ribozomální 16S genetika   MeSH
• roční období MeSH
• Varroidae mikrobiologie   fyziologie   MeSH
• včely růst a vývoj   parazitologie   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Honeybee (Apis mellifera L.) workers act as passive vectors of Paenibacillus larvae spores, which cause the quarantine disease American foulbrood (AFB). We assessed the relative proportions of P. larvae within the honeybee microbiome using metabarcoding analysis of the 16 S rRNA gene. The microbiome was analyzed in workers outside of the AFB zone (control - AFB0), in workers from asymptomatic colonies in an AFB apiary (AFB1), and in workers from colonies exhibiting clinical AFB symptoms (AFB2). The microbiome was processed for the entire community and for a cut-off microbiome comprising pathogenic/environmental bacteria following the removal of core bacterial sequences; varroosis levels were considered in the statistical analysis. No correlation was observed between AFB status and varroosis level, but AFB influenced the worker bee bacterial community, primarily the pathogenic/environmental bacteria. There was no significant difference in the relative abundance of P. larvae between the AFB1 and AFB0 colonies, but we did observe a 9-fold increase in P. larvae abundance in AFB2 relative to the abundance in AFB1. The relative sequence numbers of Citrobacter freundii and Hafnia alvei were higher in AFB2 and AFB1 than in AFB0, whereas Enterococcus faecalis, Klebsiella oxytoca, Spiroplasma melliferum and Morganella morganii were more abundant in AFB0 and AFB1 than in AFB2.

• MeSH
• analýza hlavních komponent MeSH
• biodiverzita MeSH
• diskriminační analýza MeSH
• kukla mikrobiologie   MeSH
• mikrobiota * MeSH
• Paenibacillus larvae fyziologie   MeSH
• včely mikrobiologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Blood feeding red poultry mites (RPM) serve as vectors of pathogenic bacteria and viruses among vertebrate hosts including wild birds, poultry hens, mammals, and humans. The microbiome of RPM has not yet been studied by high-throughput sequencing. RPM eggs, larvae, and engorged adult/nymph samples obtained in four poultry houses in Czechia were used for microbiome analyses by Illumina amplicon sequencing of the 16S ribosomal RNA (rRNA) gene V4 region. A laboratory RPM population was used as positive control for transcriptome analysis by pyrosequencing with identification of sequences originating from bacteria. The samples of engorged adult/nymph stages had 100-fold more copies of 16S rRNA gene copies than the samples of eggs and larvae. The microbiome composition showed differences among the four poultry houses and among observed developmental stadia. In the adults' microbiome 10 OTUs comprised 90 to 99% of all sequences. Bartonella-like bacteria covered between 30 and 70% of sequences in RPM microbiome and 25% bacterial sequences in transcriptome. The phylogenetic analyses of 16S rRNA gene sequences revealed two distinct groups of Bartonella-like bacteria forming sister groups: (i) symbionts of ants; (ii) Bartonella genus. Cardinium, Wolbachia, and Rickettsiella sp. were found in the microbiomes of all tested stadia, while Spiroplasma eriocheiris and Wolbachia were identified in the laboratory RPM transcriptome. The microbiomes from eggs, larvae, and engorged adults/nymphs differed. Bartonella-like symbionts were found in all stadia and sampling sites. Bartonella-like bacteria was the most diversified group within the RPM microbiome. The presence of identified putative pathogenic bacteria is relevant with respect to human and animal health issues while the identification of symbiontic bacteria can lead to new control methods targeting them to destabilize the arthropod host.

• MeSH
• Bacteria klasifikace   genetika   izolace a purifikace   MeSH
• bakteriální RNA genetika   MeSH
• Bartonella klasifikace   genetika   izolace a purifikace   MeSH
• druhová specificita MeSH
• mikrobiota * MeSH
• RNA ribozomální 16S genetika   MeSH
• roztoči růst a vývoj   mikrobiologie   MeSH
• taxonomické DNA čárové kódování MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika MeSH

Vysokoškolská učebnice, která se zaměřuje na různé druhy mikroorganismů a jejich ekologii a patogenitu.

• MeSH
• ekologie MeSH
• infekční nemoci MeSH
• mikrobiologické jevy MeSH
• mikrobiologie životního prostředí MeSH
• molekulární biologie MeSH
• Publikační typ
• monografie MeSH

• Konspekt
• Mikrobiologie
• Učební osnovy. Vyučovací předměty. Učebnice
• NLK Obory
• mikrobiologie, lékařská mikrobiologie
• NLK Publikační typ
• učebnice vysokých škol