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Title:Processing and mounting phlebotomine sand flies : a consensus guideline
Authors:ID Randrianambinintsoa, Fano José (Author)
ID Augendre, Laure (Author)
ID Prudhomme, Jorian (Author)
ID Martinet, Jean-Philippe (Author)
ID Loyer, Mathieu (Author)
ID Mekarnia, Nalia (Author)
ID Kerkoub, Hocine (Author)
ID Khan Perveen, Farzana (Author)
ID Huguenin, Antoine (Author)
ID Ivović, Vladimir (Author)
Files:.pdf RAZ_Randrianambinintsoa_Fano_Jose_2026.pdf (2,68 MB)
MD5: 92CC829E7A7CBE355906084859E76584
 
URL https://www.parasite-journal.org/articles/parasite/full_html/2026/01/parasite250147/parasite250147.html
 
Language:English
Work type:Article
Typology:1.02 - Review Article
Organization:FAMNIT - Faculty of Mathematics, Science and Information Technologies
UPR - University of Primorska
Abstract: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.
Keywords: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
Publication version:Version of Record
Publication date:03.04.2026
Year of publishing:2026
Number of pages:str. 1-28
Numbering:Vol. 33, article no. ǂ18
PID:20.500.12556/RUP-22904 This link opens in a new window
UDC:595.77
ISSN on article:1776-1042
DOI:10.1051/parasite/2026009 This link opens in a new window
COBISS.SI-ID:274228483 This link opens in a new window
Publication date in RUP:03.04.2026
Views:28
Downloads:1
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Record is a part of a journal

Title:Parasite
Shortened title:Parasite
Publisher:Princeps éditions
ISSN:1776-1042
COBISS.SI-ID:523860505 This link opens in a new window

Licences

License:CC BY 4.0, Creative Commons Attribution 4.0 International
Link:http://creativecommons.org/licenses/by/4.0/
Description:This is the standard Creative Commons license that gives others maximum freedom to do what they want with the work as long as they credit the author.

Secondary language

Language:French
Title:Traitement et montage des phlébotomes
Abstract:Ta članek predstavlja celovit vodnik za obdelavo in prepariranje peščenih muh, kar je ključno za identifikacijo vrst ter odkrivanje in izolacijo patogenov. Obravnava vrsto tehnik, primernih za terenske in laboratorijske razmere. Predstavljena so podrobna navodila za zbiranje, rokovanje, prepariranje ter usmrtitev peščenih muh (s priporočilom suhega zamrzovanja ali CO₂ namesto kemikalij) ter strategiji hladnega shranjevanja in konzerviranja v etanolu. Kakovost priprave določenih anatomskih struktur (spolni organi, glava in krila) je bistvena za ustrezno mikroskopsko opazovanje peščenih muh, ki je tudi podrobno predstavljeno v tem prispevku. Članek predstavlja tudi podrobno obdelavo vzorcev, vključno z uporabo reagentov, kot sta kalijev hidroksid in Marc-Andréjeva raztopina za doseganje prosojnosti preparatov. V prispevku smo primerjali različna preparirna sredstva, njihove optične lastnosti in potencial za ohranjanje preparatov. Hoyerjevo raztopino (znana tudi kot kloralna smola) zaradi učinka prosojnosti priporočamo za hitro opazovanje, zlasti spermatek, vendar ni primerna za dolgoročno shranjevanje. Med drugimi obravnavanimi sredstvi so polivinilni alkohol, Euparal® (z omejeno toleranco do vode) in kanadski balzam (v ogljikovodikih topno sredstvo), pri čemer zadnja dva sredstva omogočata dolgoročno ohranjanje. Obravnavani so tudi sodobni pristopi molekularne biologije, kot sta sekveniranje DNA in MALDI-ToF, ki zahtevajo posebno pozornost pri obdelavi vzorcev. Poleg tega so na voljo kratki videoposnetki, ki prikazujejo različne tehnike prepariranja, ter prevodi v 33 različnih jezikov, ki omogočajo, da smernice dosežejo raznolike potrebe in pričakovanja globalne znanstvene skupnosti.Cet article propose un guide complet pour le traitement et le montage des phlébotomes, étapes cruciales pour l’identification des espèces et la détection ou l’isolement des pathogènes. Il aborde diverses techniques adaptées au terrain comme au laboratoire. Le guide comprend des instructions détaillées sur la collecte, la manipulation, la protection et l’euthanasie des phlébotomes (avec recommandation de la congélation à sec ou du CO2 plutôt que de l’usage de produits chimiques), ainsi que sur les stratégies de conservation telles que la réfrigération et la conservation dans l’éthanol. La préparation des structures anatomiques spécifiques (organes génitaux, tête et ailes) pour l’observation microscopique est privilégiée dans la description des techniques de dissection. Le document présente également le traitement détaillé des échantillons, notamment la clarification à l’aide de solutions telles que la potasse puis le liquide de Marc-André. Le processus de montage compare différents milieux, en soulignant leurs propriétés optiques et leur potentiel de conservation. Le liquide de Hoyer (également appelé gomme au chloral) est recommandé pour l’observation rapide, en particulier pour les spermathèques, en raison de sa limpidité, bien qu’il ne convienne pas à la conservation à long terme. D’autres milieux de culture sont abordés, notamment l’alcool polyvinylique, l’EuparalÒ (pour une tolérance limitée à l’eau) et le baume du Canada (un milieu soluble dans les hydrocarbures), ces deux derniers permettant une conservation à long terme. Les approches innovantes de biologie moléculaire, telles que le séquençage de l’ADN et la spectrométrie de masse MALDI-ToF, qui exigent une attention particulière lors du traitement des échantillons, sont également présentées. De plus, de courtes vidéos illustrant diverses techniques de montage et des traductions en de nombreuses langues sont disponibles, permettant ainsi de répondre aux besoins et attentes variés de la communauté scientifique internationale.
Keywords:prepariranje, peščene muhe, Hoyerjeva raztopina, Marc-Andréjeva raztopina, kloralna smola, polivinilni alkohol, Euparal®, kanadski balzam, izolacija Leishmania, terenske razmere, gojenje, disekcija, molekularna biologija, MALDI-ToF, tipski primerki


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