Biopriming seeds with beneficial bacteria has potential to enhance seed germination. Therefore, in this investigation, five sulphur-oxidizing bacterial cultures, viz., Pantoea dispersa SOB2, Bacillus velezensis SN06, Bacillus cereus SN20, Bacillus tropicus SN16, and Bacillus megaterium SN11, were evaluated for different plant growth-promoting traits and their impact on Vigna radiata L. (mung bean) and Brassica juncea L. (mustard) seed germination. Among these, three bacterial cultures Pantoea dispersa SOB2, Bacillus velezensis SN06, and Bacillus megaterium SN11 evinced potential for mineral solubilization on solid medium where Pantoea dispersa SOB2 had the maximum solubilization indices-3.06, 5.14, and 2.48 for phosphate, zinc, and potassium respectively. The culture also displayed higher indole acetic acid (113.12 μg/mL), gibberellic acid (162.66 μg/mL), ammonia (5.23 μg/mL), and siderophore (69.53%) production than other bacterial cultures whereas Bacillus cereus SN20 showed maximum exopolysaccharide production (9.26 g/L). Bacterial culture Pantoea dispersa SOB2 significantly ameliorated the germination rate (3.73 no./day) and relative seed germination (208%) of Brassica juncea L., while Bacillus velezensis SN06 and Bacillus cereus SN20 followed with germination rate and relative seed germination of 2.86 no./day and 207%, respectively. Pantoea dispersa SOB2 displayed lowest mean germination time 2.91 days followed by Bacillus megaterium SN11 with 3.19 days. Biopriming with sulphur-oxidizing bacterial cultures, germination parameters of Vigna radiata L. were also markedly improved. Pantoea dispersa SOB2 demonstrated the highest germination rate (6.72 no./day), relative seed germination (115.56%), and minimum mean generation time (1.73 days). Bacillus velezensis SN06 inoculation had a beneficial effect on the seedling growth of Vigna radiata L., whereas Pantoea dispersa SOB2 greatly aided the seedling growth of Brassica juncea L. Results corroborated a prominent positive correlation between seed germination and plant growth-promoting traits. This is the first study on Pantoea dispersa as sulphur oxidizer, displaying plant growth promoting traits and seed germination potential. The potent sulphur-oxidizing bacterial cultures possessing plant growth promoting activities enhanced seed germination under in vitro conditions that could be further explored in field as biofertilizers to enhance the growth and yield of Brassica juncea L. and Vigna radiata L. crop.

• MeSH
• Bacillus * metabolism   MeSH
• Bacteria * metabolism   MeSH
• Mustard Plant * growth & development   microbiology   MeSH
• Germination * MeSH
• Indoleacetic Acids metabolism   MeSH
• Oxidation-Reduction MeSH
• Pantoea metabolism   MeSH
• Plant Growth Regulators metabolism   MeSH
• Seeds * microbiology   growth & development   MeSH
• Sulfur * metabolism   MeSH
• Vigna * growth & development   microbiology   MeSH
• Publication type
• Journal Article MeSH

Toscana virus (TOSV) is an emerging but neglected human pathogen currently circulating around the Mediterranean basin including North Africa. Human illness ranges from asymptomatic or mild flu-like syndromes to severe neurological diseases such as meningitis or meningoencephalitis. Despite its significant impact, understanding of TOSV transmission and epidemiology remains limited. Sand flies (Diptera: Phlebotominae), specifically Phlebotomus perniciosus and Phlebotomus perfiliewi, are believed to be the primary vectors of TOSV. However, the spread of TOSV to new geographical areas and its detection in other sand fly species suggest that additional species play a role in the circulation and transmission of this virus. This study investigated the vector competence of four sand fly species - P. tobbi, P. sergenti, P. papatasi, and Sergentomyia schwetzi - for two TOSV strains: 1500590 (TOSV A lineage) and MRS20104319501 (TOSV B lineage). Sand flies were orally challenged with TOSV via bloodmeals. None of the tested species showed susceptibility to the TOSV A strain. However, for TOSV B strain, P. tobbi demonstrated a high potential as a new vector, exhibiting high infection and dissemination rates. P. sergenti also showed some susceptibility to TOSV B, with the virus dissemination observed in all infected females. These finding suggests that P. tobbi and P. sergenti are new potential vectors for TOSV B. Given that P. tobbi and P. sergenti are the primary vectors of human leishmaniases in the Balkans, Turkey and Middle East, their susceptibility to TOSV could have significant epidemiological consequences. On the other hand, P. papatasi and S. schwetzi appeared refractory to TOSV B infection. Refractoriness of P. papatasi, a highly anthropophilic species distributed from the Mediterranean to the Middle East and India, suggests that this species does not contribute to TOSV circulation.

• MeSH
• Insect Vectors * virology   MeSH
• Humans MeSH
• Phlebotomus * virology   MeSH
• Psychodidae * virology   classification   MeSH
• Sandfly fever Naples virus * physiology   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Male MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

UNLABELLED: The study aimed to analyze the entomological material collected during 13 autopsies performed on the unidentified cadavers revealed at different stages of decay in the Upper Silesia Region (Poland) over 2016-2022. During the preparation of human tissues for genetic identification, we revealed larvae, puparia, and adult insects in previously undescribed locations: costal cartilage, femur nutrient canals (foramen nutrients), and tooth cavities. The taxonomical assessment was done using morphological examination or DNA barcoding, where necessary. Based on our observations, we conclude that the apical constriction, foramen, and cavities may serve as migration paths inside teeth, and the femur nutrient canals to the bone marrow. The study also revealed that the beetle Necrobia ruficollis (Fabricius, 1775) and the moth family Pyralidae Latreille, 1802 (Phycitinae) moths can form pupal chambers inside the costal cartilage, indicating that these insects can complete their life cycle inside this cache. We believe that the newly reported locations of carrion insects in human remains may be relevant to forensic entomology, as they provide new opportunities to collect insect evidence. KEY POINTS: Costal cartilage may serve as an occasional cache for adults and immatures of carrion insects.Tooth cavities and apical foramen may serve as entryways for necrophilous insect larvae.Insect larvae use nutrient canals as migratory pathways to the bone marrow.

• Publication type
• Journal Article MeSH

UNLABELLED: The Aspergillus genus encompasses a diverse array of species, some of which are opportunistic pathogens. Traditionally, human aspergillosis has primarily been linked to a few Aspergillus species, predominantly A. fumigatus. Changes in epidemiology and advancements in molecular techniques have brought attention to less common and previously unrecognized pathogenic cryptic species. Despite the taxonomic recognition of many cryptic species in section Terrei, their virulence potential and clinical implications, compared to A. terreus sensu stricto, remain poorly understood. Hence, the current study utilized the alternative in vivo model Galleria mellonella to evaluate the virulence potential of 19 accepted Aspergillus species in section Terrei, classified into three series (major phylogenetic clades): Terrei, Nivei, and Ambigui. Analyzing the median survival rates of infected larvae of all species in each series revealed that series Ambigui has a significantly lower virulence compared to series Terrei and Nivei. Taking a closer look at series Terrei and Nivei revealed a trend of survival within each clade, dividing the species into two groups: highly virulent (up to 72 h survival) and less virulent (up to 144 h survival). Histological observation, considering fungal distribution and filamentation, further supported this assessment, revealing increased distribution and hyphal formation in virulent species. Additionally, the susceptibility profile of conventional antifungals was determined, revealing an increased azole minimum inhibitory concentration for some tested cryptic species such as A. niveus and A. iranicus. Our results highlight the importance of cryptic species identification, as they can exhibit different levels of virulence and show reduced antifungal susceptibility. IMPORTANCE: With changing fungal epidemiology and an increasingly vulnerable population, cryptic Aspergillus species are emerging as human pathogens. Their diversity and clinical relevance remain underexplored, with some species showing reduced antifungal susceptibility and higher virulence, highlighting the need for better preparedness in clinical practice. Using the Galleria mellonella model, we assessed the virulence of Aspergillus species of section Terrei, including cryptic and non-cryptic species, across three series Terrei, Nivei, and Ambigui. The results revealed significant virulence variation among the series, with some cryptic species displaying high virulence. Histological analysis confirmed increased hyphal formation and fungal spread in the more virulent species. Additionally, elevated azole minimum inhibitory concentrations were also observed in certain cryptic species. This study presents novel insights into the pathogenicity of Aspergillus section Terrei, emphasizing the critical importance of accurately identifying cryptic species due to their diverse virulence potential and antifungal resistance, which may have substantial clinical implications.

• MeSH
• Antifungal Agents pharmacology   MeSH
• Aspergillus * pathogenicity   classification   drug effects   genetics   MeSH
• Aspergillosis * microbiology   MeSH
• Phylogeny MeSH
• Larva microbiology   MeSH
• Humans MeSH
• Disease Models, Animal MeSH
• Moths * microbiology   MeSH
• Virulence MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

Amphotericin B (AmB) is one of the most effective antifungal drugs, with a strong, dose-dependent activity against most Candida and Aspergillus species responsible for life-threatening infections. However, AmB is severely toxic, which hinders its broad use. In this proof-of-concept study, we demonstrate that prodrugging AmB considerably decreases AmB toxicity without affecting its fungicidal activity. For this purpose, we modified the AmB structure by attaching a designer phosphate promoiety, thereby switching off its mode of action and preventing its toxic effects. The original fungicidal activity of AmB was then restored upon prodrug activation by host plasma enzymes. These AmB prodrugs showed a safer toxicity profile than commercial AmB deoxycholate in Candida and Aspergillus species and significantly prolonged larval survival of infected Galleria mellonella larvae. Based on these findings, prodrugging toxic antifungals may be a viable strategy for broadening the antifungal arsenal, opening up opportunities for targeted prodrug design.

• MeSH
• Amphotericin B * pharmacology   MeSH
• Antifungal Agents * pharmacology   chemistry   chemical synthesis   MeSH
• Aspergillus drug effects   MeSH
• Candida drug effects   MeSH
• Larva drug effects   MeSH
• Microbial Sensitivity Tests * MeSH
• Molecular Structure MeSH
• Moths drug effects   MeSH
• Prodrugs * pharmacology   chemistry   chemical synthesis   MeSH
• Dose-Response Relationship, Drug MeSH
• Structure-Activity Relationship MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH

Bacillus thuringiensis (Bt) is known for its Cry and Vip3A pesticidal proteins with high selectivity to target pests. Here, we assessed the potential of a novel neotropical Bt strain (UFT038) against six lepidopteran pests, including two Cry-resistant populations of fall armyworm, Spodoptera frugiperda. We also sequenced and analyzed the genome of Bt UFT038 to identify genes involved in insecticidal activities or encoding other virulence factors. In toxicological bioassays, Bt UFT038 killed and inhibited the neonate growth in a concentration-dependent manner. Bt UFT038 and HD-1 were equally toxic against S. cosmioides, S. frugiperda (S_Bt and R_Cry1 + 2Ab populations), Helicoverpa zea, and H. armigera. However, larval growth inhibition results indicated that Bt UFT038 was more toxic than HD-1 to S. cosmioides, while HD-1 was more active against Chrysodeixis includens. The draft genome of Bt UFT038 showed the cry1Aa8, cry1Ac11, cry1Ia44, cry2Aa9, cry2Ab35, and vip3Af5 genes. Besides this, genes encoding the virulence factors (inhA, plcA, piplC, sph, and chi1-2) and toxins (alo, cytK, hlyIII, hblA-D, and nheA-C) were also identified. Collectively, our findings reveal the potential of the Bt UFT038 strain as a source of insecticidal genes against lepidopteran pests, including S. cosmioides and S. frugiperda.

• MeSH
• Bacillus thuringiensis * genetics   metabolism   MeSH
• Bacterial Proteins genetics   metabolism   MeSH
• Pest Control, Biological MeSH
• Endotoxins metabolism   MeSH
• Virulence Factors metabolism   MeSH
• Glycine max MeSH
• Hemolysin Proteins genetics   metabolism   pharmacology   MeSH
• Insecticides * pharmacology   metabolism   MeSH
• Larva MeSH
• Humans MeSH
• Moths * MeSH
• Infant, Newborn MeSH
• Spodoptera metabolism   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Infant, Newborn MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

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
• Genes, Essential * MeSH
• Insect Vectors genetics   MeSH
• Genes, Insect MeSH
• Larva genetics   MeSH
• Leishmania genetics   MeSH
• Phlebotomus * genetics   MeSH
• Psychodidae genetics   MeSH
• Gene Expression Profiling methods   MeSH
• Animals MeSH
• Check Tag
• Male MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

Sand flies (Diptera: Psychodidae: Phlebotominae) are blood-feeding insects that transmit the protozoan parasites Leishmania spp. and various arboviruses. The Balkan region, including the Republic of Kosovo, harbours a diverse sand fly fauna. Vector species of Leishmania infantum as well as phleboviruses are endemic; however, recent data are scarce. We performed a cross-sectional study to update the current sand fly distribution in Kosovo and assess biological as well as environmental factors associated with sand fly presence. CDC light trapping was conducted at 46 locations in 2022 and 2023, specifically targeting understudied regions in Kosovo. Individual morphological species identification was supported by molecular barcoding. The occurrence data of sand flies was used to create distribution maps and perform environmental analyses, taking elevation, wind speed and climate-related factors into account. In addition, PCR-based blood meal analysis and pathogen screening were conducted. Overall, 303 specimens of six sand fly species were trapped, predominated by Phlebotomus neglectus (97%). Barcodes from eight of nine known endemic sand fly species were obtained. Combining our data with previous surveys, we mapped the currently known sand fly distribution based on more than 4000 specimens at 177 data points, identifying Ph. neglectus and Ph. perfiliewi as the predominant species. Environmental analyses depicted two geographical groups of sand flies in Kosovo, with notable differences between the species. In total, 223 blood meals of five sand fly species were analysed. Of seven identified host species, the predominant blood meal source was observed to be cattle, but the DNA of dogs and humans, among others, was also detected. This study assessed biological as well as ecological factors of sand fly occurrence, which should help better understand and evaluate potential hot spots of disease transmission in Kosovo.

• MeSH
• Insect Vectors * physiology   parasitology   MeSH
• Leishmania infantum physiology   MeSH
• Phlebotomus * classification   physiology   parasitology   MeSH
• Cross-Sectional Studies MeSH
• Dogs MeSH
• Psychodidae physiology   parasitology   MeSH
• Animal Distribution * MeSH
• Animals MeSH
• Check Tag
• Male MeSH
• Dogs MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Geographicals
• Kosovo MeSH

Leishmania species, members of the kinetoplastid parasites, cause leishmaniasis, a neglected tropical disease, in millions of people worldwide. Leishmania has a complex life cycle with multiple developmental forms, as it cycles between a sand fly vector and a mammalian host; understanding their life cycle is critical to understanding disease spread. One of the key life cycle stages is the haptomonad form, which attaches to insect tissues through its flagellum. This adhesion, conserved across kinetoplastid parasites, is implicated in having an important function within their life cycles and hence in disease transmission. Here, we discover the kinetoplastid-insect adhesion proteins (KIAPs), which localise in the attached Leishmania flagellum. Deletion of these KIAPs impairs cell adhesion in vitro and prevents Leishmania from colonising the stomodeal valve in the sand fly, without affecting cell growth. Additionally, loss of parasite adhesion in the sand fly results in reduced physiological changes to the fly, with no observable damage of the stomodeal valve and reduced midgut swelling. These results provide important insights into a comprehensive understanding of the Leishmania life cycle, which will be critical for developing transmission-blocking strategies.

• MeSH
• Cell Adhesion MeSH
• Flagella * metabolism   MeSH
• Insect Vectors parasitology   MeSH
• Insect Proteins metabolism   genetics   MeSH
• Host-Parasite Interactions MeSH
• Leishmania * physiology   genetics   metabolism   MeSH
• Leishmaniasis parasitology   transmission   MeSH
• Protozoan Proteins metabolism   genetics   MeSH
• Psychodidae * parasitology   MeSH
• Life Cycle Stages MeSH
• Animals MeSH
• Check Tag
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

Introduction. The fungal pathogen Aspergillus fumigatus can induce prolonged colonization of the lungs of susceptible patients, resulting in conditions such as allergic bronchopulmonary aspergillosis and chronic pulmonary aspergillosis.Hypothesis. Analysis of the A. fumigatus secretome released during sub-lethal infection of G. mellonella larvae may give an insight into products released during prolonged human colonisation.Methodology.Galleria mellonella larvae were infected with A. fumigatus, and the metabolism of host carbohydrate and proteins and production of fungal virulence factors were analysed. Label-free qualitative proteomic analysis was performed to identify fungal proteins in larvae at 96 hours post-infection and also to identify changes in the Galleria proteome as a result of infection.Results. Infected larvae demonstrated increasing concentrations of gliotoxin and siderophore and displayed reduced amounts of haemolymph carbohydrate and protein. Fungal proteins (399) were detected by qualitative proteomic analysis in cell-free haemolymph at 96 hours and could be categorized into seven groups, including virulence (n = 25), stress response (n = 34), DNA repair and replication (n = 39), translation (n = 22), metabolism (n = 42), released intracellular (n = 28) and cellular development and cell cycle (n = 53). Analysis of the Gallerial proteome at 96 hours post-infection revealed changes in the abundance of proteins associated with immune function, metabolism, cellular structure, insect development, transcription/translation and detoxification.Conclusion. Characterizing the impact of the fungal secretome on the host may provide an insight into how A. fumigatus damages tissue and suppresses the immune response during long-term pulmonary colonization.

• MeSH
• Aspergillus fumigatus * metabolism   MeSH
• Aspergillosis microbiology   metabolism   MeSH
• Virulence Factors metabolism   MeSH
• Fungal Proteins * metabolism   genetics   MeSH
• Hemolymph microbiology   metabolism   MeSH
• Larva * microbiology   MeSH
• Moths * microbiology   MeSH
• Proteome analysis   MeSH
• Proteomics MeSH
• Secretome metabolism   MeSH
• Virulence MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH