Processing and mounting phlebotomine sand flies: a consensus guideline

Fano José Randrianambinintsoa; Laure Augendre; Jorian Prudhomme; Jean Philippe Martinet; Mathieu Loyer; Nalia Mekarnia; Hocine Kerkoub; Farzana Khan Perveen; Antoine Huguenin; Emilie Kariya; Mohammad Akhoundi; Andrey José De Andrade; Eduardo Berriatua; Gioia Bongiorno; Sébastien Boyer; Vasiliki Christodoulou; Magda Clara Vieira Da Costa-Ribeiro; Lucas Alexandre Farias De Souza; Huicong Ding; Blaise Dondji; Vít Dvořák; Ozge Erisoz Kasap; Eunice Aparecida Bianchi Galati; Montserrat Gállego; Cristina Ballart; Stavroula Gouzelou; Nabil Haddad; Rezki Sabrina Masse; Asrat Hailu Mekuria; Vladimir Ivovic; Szymon Kaczmarek; Mohd Khadri Shahar; Oscar D. Kirstein; Edwin Kniha; Iva Kolářová; Lincoln Timinao; Cristian Lucanas; Ognyan Mikov; Kimsear Nov; Yusuf Özbel; Bernard Pesson; Laura Cristina Posada Lopez; Didot Budi Prasetyo; Nil Rahola; Eduardo A. Rebollar-Tellez; Bruno Leite Rodrigues; Lalita Roy; Prasanta Saini; Chizu Sanjoba; Paloma Helena Fernandes Shimabukuro; Padet Siriyasatien; Agnieszka Soszyńska; Tatiana Suleşco; Massamba Sylla; Majhalia Torno; Petr Volf; Khamsing Vongphayloth; Sinh Nam Vu; April Wardhana; Eric Yessinou; Sonia Zapata; Jean Charles Gantier; Jérôme Depaquit

doi:10.1051/parasite/2026009

Processing and mounting phlebotomine sand flies: a consensus guideline

Fano José Randrianambinintsoa
, Laure Augendre
, Jorian Prudhomme
, Jean Philippe Martinet
, Mathieu Loyer
, Nalia Mekarnia
, Hocine Kerkoub
, Farzana Khan Perveen
, Antoine Huguenin
, Emilie Kariya
, Mohammad Akhoundi
, Andrey José De Andrade
, Eduardo Berriatua
, Gioia Bongiorno
, Sébastien Boyer
, Vasiliki Christodoulou
, Magda Clara Vieira Da Costa-Ribeiro
, Lucas Alexandre Farias De Souza
, Huicong Ding
, Blaise Dondji

Vít Dvořák, Ozge Erisoz Kasap, Eunice Aparecida Bianchi Galati, Montserrat Gállego, Cristina Ballart, Stavroula Gouzelou, Nabil Haddad, Rezki Sabrina Masse, Asrat Hailu Mekuria, Vladimir Ivovic, Szymon Kaczmarek, Mohd Khadri Shahar, Oscar D. Kirstein, Edwin Kniha, Iva Kolářová, Lincoln Timinao, Cristian Lucanas, Ognyan Mikov, Kimsear Nov, Yusuf Özbel, Bernard Pesson, Laura Cristina Posada Lopez, Didot Budi Prasetyo, Nil Rahola, Eduardo A. Rebollar-Tellez, Bruno Leite Rodrigues, Lalita Roy, Prasanta Saini, Chizu Sanjoba, Paloma Helena Fernandes Shimabukuro, Padet Siriyasatien, Agnieszka Soszyńska, Tatiana Suleşco, Massamba Sylla, Majhalia Torno, Petr Volf, Khamsing Vongphayloth, Sinh Nam Vu, April Wardhana, Eric Yessinou, Sonia Zapata, Jean Charles Gantier, Jérôme Depaquit

Université de Reims Champagne-Ardenne
Université Paris 13
Federal University of Parana
University of Murcia
Istituto Superiore di Sanita
Institut Pasteur du Cambodge
Laboratory for Animal Health Virology
National University of Singapore
Central Washington University
Charles University
Hacettepe University
Universidade de São Paulo
Center for Research in Environmental Epidemiology (CREAL)
University of Cyprus Medical School
American University of Beirut
Institute for Medical Research Kuala Lumpur
Addis Ababa University
University of Primorska
University of Lodz
Ministry of Health, Israel
Medical University of Vienna
Papua New Guinea Institute of Medical Research
University of the Philippines
National Center of Infectious and Parasitic Diseases Bulgaria
Ege University
Université de Strasbourg
Universidad de Antioquia
University of Montpellier
Universidad Autonoma de Nuevo Leon
B.P. Koirala Institute of Health Sciences
Indian Council of Medical Research
The University of Tokyo
Fundação Oswaldo Cruz
Chulalongkorn University
Bernhard Nocht Institute for Tropical Medicine
Laboratory Vectors & Parasites
Institut Pasteur du Laos
National Institute of Hygiene and Epidemiology Hanoi
Indonesian Agency for Agricultural Research and Development
Université d'Abomey-Calavi

Research output: Contribution to journal › Review article › peer-review

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 collection, 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 hydroxide 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, Euparal® (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.

Original language	English
Pages (from-to)	18
Number of pages	1
Journal	Parasite
Volume	33
DOIs	https://doi.org/10.1051/parasite/2026009
State	Published - 2026

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 3 Good Health and Well-being

Keywords

Canada balsam
Chloral gum
Culture
Dissection
Euparal®
Field conditions
Hoyer fluid
Leishmania isolation
MALDI-ToF
Marc-André solution
Molecular biology
Mounting
Phlebotomine sand fly
Polyvinyl alcohol
Type-specimens

Access to Document

10.1051/parasite/2026009

Cite this

Randrianambinintsoa, F. J., Augendre, L., Prudhomme, J., Martinet, J. P., Loyer, M., Mekarnia, N., Kerkoub, H., Perveen, F. K., Huguenin, A., Kariya, E., Akhoundi, M., De Andrade, A. J., Berriatua, E., Bongiorno, G., Boyer, S., Christodoulou, V., Vieira Da Costa-Ribeiro, M. C., De Souza, L. A. F., Ding, H., ... Depaquit, J. (2026). Processing and mounting phlebotomine sand flies: a consensus guideline. Parasite, 33, 18. https://doi.org/10.1051/parasite/2026009

@article{2bc71e9c67ea44718c9806911a9ac257,

title = "Processing and mounting phlebotomine sand flies: a consensus guideline",

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 collection, 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 hydroxide then Marc-Andr{\'e} 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, Euparal{\textregistered} (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 = "Canada balsam, Chloral gum, Culture, Dissection, Euparal{\textregistered}, Field conditions, Hoyer fluid, Leishmania isolation, MALDI-ToF, Marc-Andr{\'e} solution, Molecular biology, Mounting, Phlebotomine sand fly, Polyvinyl alcohol, Type-specimens",

author = "Randrianambinintsoa, \{Fano Jos{\'e}\} and Laure Augendre and Jorian Prudhomme and Martinet, \{Jean Philippe\} and Mathieu Loyer and Nalia Mekarnia and Hocine Kerkoub and Perveen, \{Farzana Khan\} and Antoine Huguenin and Emilie Kariya and Mohammad Akhoundi and \{De Andrade\}, \{Andrey Jos{\'e}\} and Eduardo Berriatua and Gioia Bongiorno and S{\'e}bastien Boyer and Vasiliki Christodoulou and \{Vieira Da Costa-Ribeiro\}, \{Magda Clara\} and \{De Souza\}, \{Lucas Alexandre Farias\} and Huicong Ding and Blaise Dondji and V{\'i}t Dvo{\v r}{\'a}k and \{Erisoz Kasap\}, Ozge and Galati, \{Eunice Aparecida Bianchi\} and Montserrat G{\'a}llego and Cristina Ballart and Stavroula Gouzelou and Nabil Haddad and Masse, \{Rezki Sabrina\} and Mekuria, \{Asrat Hailu\} and Vladimir Ivovic and Szymon Kaczmarek and Shahar, \{Mohd Khadri\} and Kirstein, \{Oscar D.\} and Edwin Kniha and Iva Kol{\'a}{\v r}ov{\'a} and Lincoln Timinao and Cristian Lucanas and Ognyan Mikov and Kimsear Nov and Yusuf {\"O}zbel and Bernard Pesson and \{Posada Lopez\}, \{Laura Cristina\} and Prasetyo, \{Didot Budi\} and Nil Rahola and Rebollar-Tellez, \{Eduardo A.\} and Rodrigues, \{Bruno Leite\} and Lalita Roy and Prasanta Saini and Chizu Sanjoba and \{Fernandes Shimabukuro\}, \{Paloma Helena\} and Padet Siriyasatien and Agnieszka Soszy{\'n}ska and Tatiana Sule{\c s}co and Massamba Sylla and Majhalia Torno and Petr Volf and Khamsing Vongphayloth and Vu, \{Sinh Nam\} and April Wardhana and Eric Yessinou and Sonia Zapata and Gantier, \{Jean Charles\} and J{\'e}r{\^o}me Depaquit",

note = "Publisher Copyright: {\textcopyright} F.J. Randrianambinintsoa et al., published by EDP Sciences, 2026.",

year = "2026",

doi = "10.1051/parasite/2026009",

language = "Ingl{\'e}s",

volume = "33",

pages = "18",

journal = "Parasite",

issn = "1252-607X",

publisher = "EDP Sciences",

}

Randrianambinintsoa, FJ, Augendre, L, Prudhomme, J, Martinet, JP, Loyer, M, Mekarnia, N, Kerkoub, H, Perveen, FK, Huguenin, A, Kariya, E, Akhoundi, M, De Andrade, AJ, Berriatua, E, Bongiorno, G, Boyer, S, Christodoulou, V, Vieira Da Costa-Ribeiro, MC, De Souza, LAF, Ding, H, Dondji, B, Dvořák, V, Erisoz Kasap, O, Galati, EAB, Gállego, M, Ballart, C, Gouzelou, S, Haddad, N, Masse, RS, Mekuria, AH, Ivovic, V, Kaczmarek, S, Shahar, MK, Kirstein, OD, Kniha, E, Kolářová, I, Timinao, L, Lucanas, C, Mikov, O, Nov, K, Özbel, Y, Pesson, B, Posada Lopez, LC, Prasetyo, DB, Rahola, N, Rebollar-Tellez, EA, Rodrigues, BL, Roy, L, Saini, P, Sanjoba, C, Fernandes Shimabukuro, PH, Siriyasatien, P, Soszyńska, A, Suleşco, T, Sylla, M, Torno, M, Volf, P, Vongphayloth, K, Vu, SN, Wardhana, A, Yessinou, E, Zapata, S, Gantier, JC & Depaquit, J 2026, 'Processing and mounting phlebotomine sand flies: a consensus guideline', Parasite, vol. 33, pp. 18. https://doi.org/10.1051/parasite/2026009

TY - JOUR

T1 - Processing and mounting phlebotomine sand flies

T2 - a consensus guideline

AU - Randrianambinintsoa, Fano José

AU - Augendre, Laure

AU - Prudhomme, Jorian

AU - Martinet, Jean Philippe

AU - Loyer, Mathieu

AU - Mekarnia, Nalia

AU - Kerkoub, Hocine

AU - Perveen, Farzana Khan

AU - Huguenin, Antoine

AU - Kariya, Emilie

AU - Akhoundi, Mohammad

AU - De Andrade, Andrey José

AU - Berriatua, Eduardo

AU - Bongiorno, Gioia

AU - Boyer, Sébastien

AU - Christodoulou, Vasiliki

AU - Vieira Da Costa-Ribeiro, Magda Clara

AU - De Souza, Lucas Alexandre Farias

AU - Ding, Huicong

AU - Dondji, Blaise

AU - Dvořák, Vít

AU - Erisoz Kasap, Ozge

AU - Galati, Eunice Aparecida Bianchi

AU - Gállego, Montserrat

AU - Ballart, Cristina

AU - Gouzelou, Stavroula

AU - Haddad, Nabil

AU - Masse, Rezki Sabrina

AU - Mekuria, Asrat Hailu

AU - Ivovic, Vladimir

AU - Kaczmarek, Szymon

AU - Shahar, Mohd Khadri

AU - Kirstein, Oscar D.

AU - Kniha, Edwin

AU - Kolářová, Iva

AU - Timinao, Lincoln

AU - Lucanas, Cristian

AU - Mikov, Ognyan

AU - Nov, Kimsear

AU - Özbel, Yusuf

AU - Pesson, Bernard

AU - Posada Lopez, Laura Cristina

AU - Prasetyo, Didot Budi

AU - Rahola, Nil

AU - Rebollar-Tellez, Eduardo A.

AU - Rodrigues, Bruno Leite

AU - Roy, Lalita

AU - Saini, Prasanta

AU - Sanjoba, Chizu

AU - Fernandes Shimabukuro, Paloma Helena

AU - Siriyasatien, Padet

AU - Soszyńska, Agnieszka

AU - Suleşco, Tatiana

AU - Sylla, Massamba

AU - Torno, Majhalia

AU - Volf, Petr

AU - Vongphayloth, Khamsing

AU - Vu, Sinh Nam

AU - Wardhana, April

AU - Yessinou, Eric

AU - Zapata, Sonia

AU - Gantier, Jean Charles

AU - Depaquit, Jérôme

PY - 2026

Y1 - 2026

N2 - 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 collection, 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 hydroxide 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, Euparal® (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.

AB - 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 collection, 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 hydroxide 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, Euparal® (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.

KW - Canada balsam

KW - Chloral gum

KW - Culture

KW - Dissection

KW - Euparal®

KW - Field conditions

KW - Hoyer fluid

KW - Leishmania isolation

KW - MALDI-ToF

KW - Marc-André solution

KW - Molecular biology

KW - Mounting

KW - Phlebotomine sand fly

KW - Polyvinyl alcohol

KW - Type-specimens

UR - https://www.scopus.com/pages/publications/105034952687

U2 - 10.1051/parasite/2026009

DO - 10.1051/parasite/2026009

M3 - Artículo de revisión

C2 - 41930829

AN - SCOPUS:105034952687

SN - 1252-607X

VL - 33

SP - 18

JO - Parasite

JF - Parasite

ER -

Processing and mounting phlebotomine sand flies: a consensus guideline

Abstract

UN SDGs

Keywords

Access to Document

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