1. Processing and mounting phlebotomine sand flies : a consensus guidelineFano José Randrianambinintsoa, Laure Augendre, Jorian Prudhomme, Jean-Philippe Martinet, Mathieu Loyer, Nalia Mekarnia, Hocine Kerkoub, Farzana Khan Perveen, Antoine Huguenin, Vladimir Ivović, 2026, pregledni znanstveni članek Opis: This article provides a comprehensive guide for the processing and mounting of phlebotomine sand fly specimens, which is crucial for species identification and pathogen detection and isolation. It discusses a range of techniques suitable for both field and laboratory settings. The guide includes detailed instructions on sand fly collec- tion, handling, covering, and euthanasia (recommending dry freezing or CO2 over chemicals) as well as conservation strategies, such as cold storage and preservation in ethanol. The quality of preparation of certain anatomical structures (genital organs, head and wings) is essential for their proper microscopic observation and is described in this work. The article also presents detailed sample processing, including the clearing process with agents such as potassium hydrox- ide then Marc-André solution. The mounting process compares different media, emphasizing their optical properties and preservation potential. Hoyer fluid (also known as chloral gum) is recommended for quick observation, particularly for spermathecae, due to its clarity, although it is not suitable for long-term storage. Other media discussed include polyvinyl alcohol, EuparalÒ (for limited water tolerance), and Canada balsam (a hydrocarbon-soluble medium), with the latter two offering long-term preservation capabilities. Innovative molecular biology approaches such as DNA sequencing and MALDI-ToF, which require particular attention to sample processing, are also addressed. Furthermore, short video clips illustrating various mounting techniques as well as translations in many different languages are provided, allowing the guideline to reach the diverse needs and expectations of the global scientific community.
Ključne besede: mounting, Phlebotomine sand fly, Hoyer fluid, Marc-André solution, Chloral gum, Polyvinyl alcohol, Euparal®, Canada balsam, Leishmania isolation, field conditions, culture, dissection, molecular biology, MALDI-ToF, type-specimens
Objavljeno v RUP: 03.04.2026; Ogledov: 630; Prenosov: 11
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2. First molecular evidence of Leishmania parasites in sand flies (Diptera: Phlebotominae) from SloveniaGioia Bongiorno, Katja Adam, Ilaria Bernardini, Claudia Mangiapelo, Eleonora Fiorentino, Trentina Di Muccio, Vladimir Ivović, 2025, izvirni znanstveni članek Opis: Background Sand flies (Diptera: Phlebotominae) are vectors of Leishmania spp., protozoan parasites that cause leish- maniasis, a zoonosis endemic in the Mediterranean region. Although Slovenia is not considered endemic, its proximity to affected areas and the presence of competent vectors underscore the importance of entomological surveillance. As part of the CLIMOS project, we investigated sand fly species composition, seasonal abundance, and the presence of Leishmania parasites at two sites in southwestern Slovenia: Cetore and Velike Žablje. Methods From May to October 2023, adult sand flies were collected using Centers for Disease Control and Preven- tion (CDC) miniature light traps. Specimens were morphologically identified and screened for Leishmania spp. DNA using quantitative polymerase chain reaction (PCR) targeted kinetoplast DNA, followed by species confirmation with ITS-1 nested PCR and restriction fragment length polymorphism (RFLP) analysis. Results A total of 274 sand flies were collected, predominantly Phlebotomus neglectus, P. perniciosus, and P. mascittii. Abundance peaked in July in Cetore and August in Velike Žablje, likely reflecting local climatic differences. Among 25 pools of female sand flies analyzed, Leishmania DNA was detected in three pools (12.0%), specifically in P. neglectus and P. mascittii. The cycle threshold values (Ct 35–37) indicated low parasite DNA loads. ITS-1 amplification was unsuc- cessful, likely due to low DNA concentration. Conclusions This study provides the first molecular evidence of Leishmania DNA in sand flies from Slovenia. The detection of Leishmania in P. mascittii, a species with uncertain vector competence, is particularly noteworthy and warrants further investigation. Although species identification of the parasite was not possible, the findings suggest potential for autochthonous transmission and highlight the need for continued surveillance and research in the region.
Ključne besede: sand flies, Leishmania, Slovenia, vector surveillance, seasonality
Objavljeno v RUP: 28.08.2025; Ogledov: 1060; Prenosov: 7
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4. Re-emergence of visceral and cutaneous leishmaniasis in the Greek island of CreteVasiliki Christodoulou, Maria Antoniou, Pantelis Ntais, Ippokratis Messaritakis, Vladimir Ivović, Jean-Pierre Dedet, Francine Pratlong, Vít Dvořák, Yiannis Tselentis, 2012, izvirni znanstveni članek Ključne besede: Leishmania tropica MON-300, L. infantum, Phlebotomine sand flies, Crete
Objavljeno v RUP: 15.10.2015; Ogledov: 4579; Prenosov: 121
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