We investigated gene expression patterns in Lutzomyia and Phlebotomus sand fly vectors of leishmaniases. Using quantitative PCR, we assessed the expression stability of potential endogenous control genes commonly used in dipterans. We analyzed Lutzomyia longipalpis and Phlebotomus papatasi samples from L3 and L4 larval stages, adult sand flies of different sexes, diets, dsRNA injection, and Leishmania infection. Six genes were evaluated: actin, α-tubulin, GAPDH, 60 S ribosomal proteins L8 and L32 (RiboL8 and RiboL32), and elongation factor 1-α (EF1-α). EF1-α was among the most stably expressed along with RiboL8 in L. longipalpis larvae and RiboL32 in adults. In P. papatasi, EF1-α and RiboL32 were the top in larvae, while EF1-α and actin were the most stable in adults. RiboL8 and actin were the most stable genes in dissected tissues and infected guts. Additionally, five primer pairs designed for L. longipalpis or P. papatasi were effective in PCR with Lutzomyia migonei, Phlebotomus duboscqi, Phlebotomus perniciosus, and Sergentomyia schwetzi cDNA. Furthermore, L. longipalpis RiboL32 and P. papatasi α-tubulin primers were suitable for qPCR with cDNA from the other four species. Our research provides tools to enhance relative gene expression studies in sand flies, facilitating the selection of endogenous control for qPCR.

• MeSH
• esenciální geny * MeSH
• hmyz - vektory genetika   MeSH
• hmyzí geny MeSH
• larva genetika   MeSH
• Leishmania genetika   MeSH
• Phlebotomus * genetika   MeSH
• Psychodidae genetika   MeSH
• stanovení celkové genové exprese metody   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Equids may be infected by zoonotic Leishmania spp., including Leishmania infantum, in regions where canine leishmaniasis (CanL) is endemic, and Leishmania martiniquensis, which has been reported in horses from Central Europe. This study was designed to evaluate the occurrence of both Leishmania spp. among equids living in CanL endemic areas of Italy, as well as to identify dipteran vectors from the same habitats. From March to October 2023, blood, serum and tissue samples from skin lesions were collected from equids (n = 98; n = 56 donkeys and n = 42 horses) living in Italy, as well as sand flies and biting midges. Blood samples (n = 98) and skin lesions (n = 56) were tested for Leishmania spp. by conventional and real time PCRs and sera were tested by immunofluorescence antibody tests (IFAT) for both L. infantum and L. martiniquensis. Insects were morphologically identified, and female specimens (n = 268 sand flies, n = 7 biting midges) analyzed for Leishmania DNA, as well as engorged sand flies (n = 16) for blood-meal detection. Two animals with skin lesions (i.e., one donkey and one horse) scored positive for Leishmania spp. DNA, and 19 animals (i.e., 19.4%; n = 13 donkeys and n = 6 horses) were seropositive for L. infantum, with five of them also for L. martiniquensis. Most seropositive animals had no dermatological lesions (i.e., 68.4%) while both animals molecularly positive for Leishmania spp. scored seronegative. Of the 356 sand flies collected, 12 females (i.e., n = 8 Sergentomyia minuta; n = 3 Phlebotomus perniciosus, n = 1 Phlebotomus perfiliewi) were positive for Leishmania spp. DNA, and one out of seven biting midges collected was DNA-positive for L. infantum. Moreover, engorged sand flies scored positive for human and equine DNA. Data suggest that equids living in CanL endemic areas are exposed to Leishmania spp., but their role in the circulation of the parasite needs further investigations.

• MeSH
• Ceratopogonidae parazitologie   MeSH
• endemické nemoci veterinární   MeSH
• Equidae * parazitologie   MeSH
• hmyz - vektory * parazitologie   MeSH
• koně parazitologie   MeSH
• Leishmania infantum izolace a purifikace   genetika   MeSH
• Leishmania * izolace a purifikace   genetika   klasifikace   MeSH
• leishmanióza * veterinární   epidemiologie   parazitologie   přenos   MeSH
• nemoci koní parazitologie   epidemiologie   MeSH
• nemoci psů * parazitologie   epidemiologie   přenos   MeSH
• psi MeSH
• Psychodidae parazitologie   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• psi MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Itálie MeSH

Necrophagous Diptera are the most important group of insects used for the purposes of forensic entomology. While the most utilized fly family in this context is the family Calliphoridae, there are several other families that can be of great importance during real-case investigations. This article analyzes the necrophagous flies of all families recorded from 160 real cases in Switzerland between 1993 and 2007. A total of 56 species belonging to 16 families was identified with Calliphoridae being the most dominant family (90.63% of all cases), followed by Muscidae (26.25%), Sarcophagidae (19.38%), Phoridae (14.38%), and Fanniidae (12.50%). For specimens that were difficult to identify morphologically, a new PCR primer has been specifically designed for the amplification of a short, informative COI barcode in degraded museum samples of forensically important Diptera taxa. The richest family in terms of species was the family Muscidae with 16 species. Fannia fuscula (Fallen) and Fannia monilis (Haliday) were recorded from human cadavers for the first time. The study highlights the importance of different fly families in forensic investigation, enhancing our comprehension of their prevalence and dispersion in real cases in Central Europe. The results pave the way for additional exploration, especially regarding the involvement of less frequently observed species in forensic entomology.

• MeSH
• Calliphoridae MeSH
• Diptera * MeSH
• entomologie MeSH
• lidé MeSH
• Muscidae * MeSH
• Sarcophagidae * MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH
• Geografické názvy
• Švýcarsko MeSH

In the family of fruit bats, Pteropodidae Gray, 1821, as in the third most diverse group of bats (Chiroptera), the bacterium of the genus Bartonella was detected in several species as well as in a few species of their insect ectoparasites in some tropical and sub-tropical regions of the Old World. The Egyptian fruit bat, Rousettus aegyptiacus (Geoffroy, 1810), is one of the most widespread fruit bats, occurring between South Africa, Senegal, and Pakistan. In this bat species, Candidatus Bartonella rousetti has been detected in three African populations in Nigeria, Kenya, and Zambia. This fruit bat, however, also occurs in the Palaearctic, an area isolating the species geographically and phylogenetically from the Afrotropical part of its distribution range. We screened the blood-sucking bat flies (family Nycteribiidae) from R. aegyptiacus for the presence of the Bartonella bacteria. A rich material of bat fly Eucampsipoda aegyptia (Macquart, 1850), a monoxenous ectoparasite of the Egyptian fruit bats, was collected at 26 localities in seven countries (Egypt, Iran, Jordan, Lebanon, Oman, United Arab Emirates, and Yemen) of the Middle East in 2007-2013. The DNA isolates from the bat flies were subjected to a three-marker (gltA, ssrA, and intergenic spacer region, ITS) multilocus sequence analysis. Based on the amplification of the fragment of ssrA gene by a real-time PCR, 65 E. aegyptia samples from 19 localities in all seven countries were positive for the bacteria. One to five Bartonella-positive individuals of E. aegyptia were collected per one individual of R. aegyptiacus. An analysis of the ITS and gltA genes indicated the presence of an uncultured Bartonella sp., belonging to the Cand. B. rousetti genogroup, identified from populations of the Egyptian fruit bat in Africa. These results support the hypothesis that Bartonella's diversity corresponds to its host's diversity (and phylogenetic structure). Specific lineages of pathogens are present in specific phylogenetic groups of bats.

• MeSH
• Bartonella * genetika   MeSH
• Chiroptera * MeSH
• fylogeneze MeSH
• intergenová DNA MeSH
• lidé MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Keňa MeSH
• Střední východ MeSH

A novel Bartonella-like symbiont (BLS) of Tyrophagus putrescentiae was characterized. BLS formed a separate cluster from the Bartonella clade together with an ant symbiont. BLS was present in mite bodies (103 16S DNA copies/mite) and feces but was absent in eggs. This indicated the presence of the BLS in mite guts. The BLS showed a reduction in genome size (1.6 Mb) and indicates gene loss compared to Bartonella apis. The BLS can be interacted with its host by using host metabolic pathways (e.g., the histidine and arginine metabolic pathways) as well as by providing its own metabolic pathways (pantothenate and lipoic acid) to the host, suggesting the existence of a mutualistic association. Our experimental data further confirmed these potential mutualistic nutritional associations, as cultures of T. putrescentiae with low BLS abundance showed the strongest response after the addition of vitamins. Despite developing an arguably tight dependency on its host, the BLS has probably retained flagellar mobility, as evidenced by the 32 proteins enriched in KEGG pathways associated with flagellar assembly or chemotaxis (e.g., fliC, flgE, and flgK, as highly expressed genes). Some of these proteins probably also facilitate adhesion to host gut cells. The microcin C transporter was identified in the BLS, suggesting that microcin C may be used in competition with other gut bacteria. The 16S DNA sequence comparison indicated a mite clade of BLSs with a broad host range, including house dust and stored-product mites. Our phylogenomic analyses identified a unique lineage of arachnid specific BLSs in mites and scorpions.IMPORTANCEA Bartonella-like symbiont was found in an astigmatid mite of allergenic importance. We assembled the genome of the bacterium from metagenomes of different stored-product mite (T. putrescentiae) cultures. The bacterium provides pantothenate and lipoic acid to the mite host. The vitamin supply explains the changes in the relative abundance of BLSs in T. putrescentiae as the microbiome response to nutritional or pesticide stress, as observed previously. The phylogenomic analyses of available 16S DNA sequences originating from mite, scorpion, and insect samples identified a unique lineage of arachnid specific forming large Bartonella clade. BLSs associated with mites and a scorpion. The Bartonella clade included the previously described Ca. Tokpelaia symbionts of ants.

• MeSH
• Acaridae * mikrobiologie   MeSH
• alergeny MeSH
• Bacteria MeSH
• Bartonella * genetika   MeSH
• kyselina lipoová * MeSH
• roztoči * genetika   MeSH
• symbióza MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

The corpora allata-corpora cardiaca (CA-CC) is an endocrine gland complex that regulates mosquito development and reproduction through the synthesis of juvenile hormone (JH). Epoxidase (Epox) is a key enzyme in the production of JH. We recently utilized CRISPR/Cas9 to establish an epoxidase-deficient (epox-/-) Aedes aegypti line. The CA from epox-/- mutants do not synthesize epoxidated JH III but methyl farneosate (MF), a weak agonist of the JH receptor, and therefore have reduced JH signalling. Illumina sequencing was used to examine the differences in gene expression between the CA-CC from wild type (WT) and epox-/- adult female mosquitoes. From 18,034 identified genes, 317 were significantly differentially expressed. These genes are involved in many biological processes, including the regulation of cell proliferation and apoptosis, energy metabolism, and nutritional uptake. In addition, the same CA-CC samples were also used to examine the microRNA (miRNA) profiles of epox-/- and WT mosquitoes. A total of 197 miRNAs were detected, 24 of which were differentially regulated in epox-/- mutants. miRNA binding sites for these particular miRNAs were identified using an in silico approach; they target a total of 101 differentially expressed genes. Our results suggest that a lack of epoxidase, besides affecting JH synthesis, results in the diminishing of JH signalling that have significant effects on Ae. aegypti CA-CC transcriptome profiles, as well as its miRNA repertoire.

• MeSH
• Aedes * genetika   metabolismus   MeSH
• corpora allata metabolismus   MeSH
• exprese genu MeSH
• juvenilní hormony metabolismus   MeSH
• mikro RNA * genetika   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Cimex lectularius, known as the common bed bug, is a widespread hematophagous human ectoparasite and urban pest that is not known to be a vector of any human infectious disease agents. However, few studies in the era of molecular biology have profiled the microorganisms harbored by field populations of bed bugs. The objective of this study was to examine the viruses present in a large sampling of common bed bugs and related bat bugs (Cimex pipistrelle). RNA sequencing was undertaken on an international sampling of > 500 field-collected bugs, and multiple workflows were used to assemble contigs and query these against reference nucleotide databases to identify viral genomes. Shuangao bed bug virus 2, an uncharacterized rhabdovirus previously discovered in Cimex hemipterus from China, was found in several bed bug pools from the USA and Europe, as well as in C. pipistrelle, suggesting that this virus is common among bed bug populations. In addition, Shuangao bed bug virus 1 was detected in a bed bug pool from China, and sequences matching Enterobacteria phage P7 were found in all bed bug pools, indicating the ubiquitous presence of phage-derived elements in the genome of the bed bug or its enterobacterial symbiont. However, viral diversity was low in bed bugs in our study, as no other viral genomes were detected with significant coverage. These results provide evidence against frequent virus infection in bed bugs. Nonetheless, our investigation had several important limitations, and additional studies should be conducted to better understand the prevalence and composition of viruses in bed bugs. Most notably, our study largely focused on insects from urban areas in industrialized nations, thus likely missing infrequent virus infections and those that could occur in rural or tropical environments or developing nations.

• MeSH
• infestace ektoparazity * MeSH
• lidé MeSH
• štěnice * genetika   MeSH
• viry * genetika   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Čína MeSH
• Evropa MeSH

Úvod: Alergia na jed blanokrídleho hmyzu je druhou najčastejšou príčinou anafylaxie detského veku. V súčasnosti jediná kauzálna terapia, ktorá môže navodiť toleranciu, je venómová imunoterapia (VIT). Ciele: V našej práci sme sa zamerali na analýzu vybraných klinických a laboratórnych charakteristík v skupine pediatrických pacientov s alergiou na jed blanokrídleho hmyzu liečených VIT. Materiál a metódy: Vytvorili sme prospektívnu štúdiu, do ktorej bolo postupne zaradených 20 detských pacientov (13 chlapcov, 7 dievčat), ktorí spĺňali indikačné kritéria pre VIT. Dáta boli zberané od roku 2015 do roku 2022. Definovali sme základnú charakteristiku súboru, zamerali sme sa na celkovú toleranciu liečby u pediatrických pacientov a výskyt vedľajších účinkov počas podávania VIT, taktiež sme vykonali odber krvi na stanovanie dynamiky laboratórnych imunologických parametrov v presne stanovených časových intervaloch. Výsledky: Priemerný vek bol 11 ± 4,37 rokov, pričom priemerný vek prvej systémovej reakcie po bodnutí bol 9,15 ± 3,58 rokov. Z 20 zaradených pacientov bolo 14 alergických na jed včely a 6 na jed osy. VIT bola priemerne začatá vo veku 10 ± 3,86 rokov, z toho 2 pacienti ju začali ešte pred dovŕšením piateho roku života. Medzi najčastejšie zastúpené klinické prejavy systémovej reakcie v našom súbore patrili angioedém tváre (70 %), dyspnoe (65 %), urtikária (55 %) a celotelový pruritus (45 %). Výskyt vedľajších príhod po VIT sme najčastejšie pozorovali počas úvodnej fázy podávania v zmysle lokálnych reakcií, pričom u žiadneho pacienta nedošlo k rozvoju systémovej reakcie. Prirodzená reexpozícia počas VIT sa vyskytla u 25 % pacientov, pričom u žiadneho sa nerozvinula systémová reakcia. V priebehu VIT sme zaznamenali aj dynamické zmeny jednotlivých hodnotených laboratórnych parametrov s postupným poklesom špecifických IgE proti extraktu a alergénovým komponentom hmyzieho jedu a vzostup špecifických IgG4 proti alergénovému extraktu. Záver: Alergiu na jed blanokrídleho hmyzu radíme medzi závažný až život ohrozujúci stav, pričom v súčasnosti jediná možná terapia, ktorou môžeme predchádzať vzniku systémovej reakcie a zlepšiť kvalitu života pacientom je VIT. U detských pacientov je udávaná vyššia účinnosť VIT a taktiež nižšie riziko zlyhania liečby. Naše skúsenosti dokladujú účinnosť aj bezpečnosť VIT v detskom veku a zároveň potvrdzujú pozitívne zmeny v laboratórnych parametroch naznačujúce indukciu imunitnej tolerancie.

Introduction: Hymenoptera venom allergy is second most common cause of anaphylaxis in childhood. Currenty, the only causal therapy that can induce tolerance is venom immunotherapy (VIT). Aim: In our work, we focused on the analysis of selected clinical and laboratory characteristics in a group of pediatric patients with hymenoptera venom allergy treated with VIT. Material and methods: We created a prospective study in which we gradually included 20 pediatric patients (13 boys, 7 girls), which fulfilled indication criteria for VIT. Data were collected from 2015 to 2022. We defined the basic characteristics of the set, we focused on the overall tolerance of treatment in pediatric patients and the occurance of adverse effects during VIT administration, we also performed blood sampling to determine the dynamics of immunological laboratory parameters in precisely determined time intervals. Results: The mean age was 11 ± 4,37 years, with the mean age of firts systemic reactions after the hymenoptera sting 9,15 ± 3,58 years. Of the 20 enrolled patients, 14 were allergic to bee venom and 6 to waps venom. VIT was started at an average age of 10 ± 3,86 years, of which 2 patients started before the fifth year of life. The most frequently represented clinical manifestations of systemic reaction in our group were facial angioedema (70 %), dyspnea (65 %), urticaria (55 %) and whole body pruritus (45 %). We observed the occurence of the adverse effects most often during the initial phase of administration in terms of local reactions, whereas no patient developed a systemic reaction. Spontaneus re-exposure during VIT occured in 25 % of patients, whereas no patient developed a systemic reaction. During VIT we also noted dynamic changes in individual evaluated laboratory parameters with a gradual decrease of specific IgE against the extract and allergen component of hymenoptera venom and increase of specific IgG4 against the allergen extract. Conclusion: Allergy to hymenoptera venom is classified as a serious to life-threating condition, while the only possible therapy nowadays that can prevent systemic reaction and improve the patient´s quality of life is VIT. A higher effectiveness of VIT and also a lower risk of treatment failure are described in pediatric patients. Our experencie clearly proves the effectiveness and safety of VIT in childhood and at the same time confirms positive changes in laboratory parameters indicating the induction of immune tolerance.

• MeSH
• alergie terapie   MeSH
• imunoterapie metody   MeSH
• kousnutí a bodnutí hmyzem * patologie   terapie   MeSH
• lidé MeSH
• mladiství MeSH
• předškolní dítě MeSH
• prospektivní studie MeSH
• včelí jedy škodlivé účinky   terapeutické užití   MeSH
• vosí jedy škodlivé účinky   terapeutické užití   MeSH
• Check Tag
• lidé MeSH
• mladiství MeSH
• mužské pohlaví MeSH
• předškolní dítě MeSH
• ženské pohlaví MeSH

INTRODUCTION: N-glycosylation is a ubiquitous and variable posttranslational modification that regulates physiological functions of secretory and membrane-associated proteins and the dysregulation of glycosylation pathways is often associated with cancer growth and metastasis. Prostate-specific membrane antigen (PSMA) is an established biomarker for prostate cancer imaging and therapy. METHODS: Mass spectrometry was used to analyze the distribution of the site-specific glycoforms of PSMA in insect, human embryonic kidney, and prostate cancer cells, and in prostate tissue upon immunoaffinity enrichment. RESULTS: While recombinant PSMA expressed in insect cells was decorated mainly by paucimannose and high mannose glycans, complex, hybrid, and high mannose glycans were detected in samples from human cells and tissue. We noted an interesting spatial distribution of the glycoforms on the PSMA surface-high mannose glycans were the dominant glycoforms at the N459, N476, and N638 sequons facing the plasma membrane, while the N121, N195, and N336 sites, located at the exposed apical PSMA domain, carried primarily complex glycans. The presence of high mannose glycoforms at the former sequons likely results from the limited access of enzymes of the glycosynthetic pathway required for the synthesis of the complex structures. In line with the limited accessibility of membrane-proximal sites, no glycosylation was observed at the N51 site positioned closest to the membrane. CONCLUSIONS: Our study presents initial descriptive analysis of the glycoforms of PSMA observed in cell lines and in prostate tissue. It will hopefully stimulate further research into PSMA glycoforms in the context of tumor staging, noninvasive detection of prostate tumors, and the impact of glycoforms on physicochemical and enzymatic characteristics of PSMA in a tissue-specific manner.

• MeSH
• antigeny povrchové metabolismus   MeSH
• buněčné linie MeSH
• glutamátkarboxypeptidasa II metabolismus   MeSH
• glykosylace MeSH
• hmotnostní spektrometrie metody   MeSH
• lidé MeSH
• nádorové biomarkery analýza   MeSH
• nádory prostaty * metabolismus   patologie   MeSH
• polysacharidy * klasifikace   metabolismus   MeSH
• posttranslační úpravy proteinů MeSH
• prostata * enzymologie   metabolismus   patologie   MeSH
• staging nádorů MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH

Anthropogenic habitats that are contaminated by toxic elements were recently shown to host abundant and diverse assemblages of bees and wasps (Hymenoptera: Aculeata), including numerous threatened species. However, toxic elements adversely affect insect fitness. We address the effects of toxic elements on aculeate inquilines that occupy Lipara lucens-induced galls on the common reed, Phragmites australis. We hypothesized that contamination of potential nesting and feeding habitats is associated with adverse changes in bee and wasp populations that are attracted in these environments. To address this hypothesis, we analyzed the contents of As, Cd, Cu, Pb, Zn, Fe, and S in site-matched samples of soil, reed galls, and crabronid wasp bodies and correlated them with abundance and species richness of aculeate hymenopterans in reed galls and with the number of larvae in nests of the eudominant hymenopteran, Pemphredon fabricii. The common reed was present at all the examined sites, and L. lucens-induced galls were present at all but one sampling site; the single exception was the sampling site with the highest contents of four of the seven analyzed elements. The alpha diversity of gall-associated aculeate inquilines, abundance of P. fabricii, and number of prepupae per nest of P. fabricii were not correlated with the contents of any of the seven analyzed toxic elements. We found P. fabricii to be abundantly present in habitats with extreme concentrations of toxic elements. Exposed P. fabricii accumulated Cd, Cu, and Pb, while they eliminated Fe and Zn. The obtained data did not support the hypothesis that heavy metal contamination of anthropogenic sites affects P. fabricii and other reed gall-associated aculeates.

• MeSH
• Diptera * MeSH
• ekosystém MeSH
• hmyz MeSH
• larva MeSH
• sršňovití * MeSH
• včely MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH