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      Laboratory colonization and mass rearing of phlebotomine sand flies (Diptera, Psychodidae) Translated title: Établissement de colonies de laboratoire et élevage de masse des phlébotomes (Diptera, Psychodidae)

      1 , * , 2 , 3 , 3 , 4

      Parasite

      EDP Sciences

      Phlebotomus, Lutzomyia, sand fly colony, leishmaniasis, mass rearing

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          Abstract

          Laboratory colonies of phlebotomine sand flies are necessary for experimental study of their biology, behaviour and mutual relations with disease agents and for testing new methods of vector control. They are indispensable in genetic studies and controlled observations on the physiology and behaviour of sand flies, neglected subjects of high priority. Colonies are of particular value for screening insecticides. Colonized sand flies are used as live vector models in a diverse array of research projects, including xenodiagnosis, that are directed toward control of leishmaniasis and other sand fly-associated diseases. Historically, labour-intensive maintenance and low productivity have limited their usefulness for research, especially for species that do not adapt well to laboratory conditions. However, with growing interest in leishmaniasis research, rearing techniques have been developed and refined, and sand fly colonies have become more common, enabling many significant breakthroughs. Today, there are at least 90 colonies representing 21 distinct phlebotomine sand fly species in 35 laboratories in 18 countries worldwide. The materials and methods used by various sand fly workers differ, dictated by the availability of resources, cost or manpower constraints rather than choice. This paper is not intended as a comprehensive review but rather a discussion of methods and techniques most commonly used by researchers to initiate, establish and maintain sand fly colonies, with emphasis on the methods proven to be most effective for the species the authors have colonized. Topics discussed include collecting sand flies for colony stock, colony initiation, maintenance and mass-rearing procedures, and control of sand fly pathogens in colonies.

          Translated abstract

          Les colonies de laboratoire de phlébotomes sont nécessaires pour une étude expérimentale de leur biologie, leur comportement et leurs relations mutuelles avec des agents pathogènes et pour tester de nouvelles méthodes de lutte antivectorielle. Elles sont indispensables dans les études génétiques et les observations contrôlées sur la physiologie et le comportement des phlébotomes, sujets négligés de haute priorité. Les colonies ont une valeur particulière pour le criblage des insecticides. Les phlébotomes en élevage sont utilisés comme modèles de vecteurs vivants dans un éventail varié de projets de recherche, y compris le xénodiagnostic, qui visent le contrôle de la leishmaniose et d'autres maladies associées aux phlébotomes. Historiquement, la maintenance à forte intensité de main-d'œuvre et la faible productivité ont limité leur utilité pour la recherche, en particulier pour les espèces qui ne s'adaptent pas bien aux conditions de laboratoire. Mais, avec un intérêt croissant pour la recherche sur la leishmaniose, les techniques d'élevage ont été développées et affinées, et les colonies de phlébotomes sont devenues plus fréquentes, permettant de nombreuses percées significatives. Aujourd'hui, il y a au moins 90 colonies représentant 21 espèces distinctes de phlébotomes dans 35 laboratoires et 18 pays à travers le monde. Les matériaux et les méthodes utilisés par divers chercheurs sur les phlébotomes diffèrent, dictés par la disponibilité des ressources et les contraintes de coûts ou de main-d'œuvre plutôt que par le choix. Cet article n'est pas destiné à être un examen complet, mais plutôt une discussion sur les méthodes et les techniques les plus utilisées par les chercheurs pour initier, établir et maintenir les colonies de phlébotomes, en mettant l'accent sur les méthodes démontrées les plus efficaces pour les espèces que les auteurs ont établies en colonies. Les sujets abordés comprennent la collecte de phlébotomes pour les stocks de colonies, l'initiation des colonies, les procédures de maintenance et d'élevage et le contrôle des agents pathogènes des phlébotomes dans les colonies.

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          Most cited references 61

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          Establishment and maintenance of sand fly colonies.

           P Volf,  V Volfova (2011)
          Sand flies used to have a reputation for being difficult and labour-intensive to breed. Here we summarize our experience with establishment and maintenance of sand fly colonies and their use for infective experiments: techniques for collection and handling wild-caught females, rearing larvae and adults and experimental infections of sand flies by Leishmania using membrane feeding. In addition, we compare major life cycle parameters between various colonies maintained under standard laboratory conditions. The sand fly rearing is tricky but some species can be reared in large numbers with a minimum of space and equipment. Initiation of new colonies from endemic sites is a prerequisite for accurate studies on parasite-vector interaction but it is more difficult step than routine maintenance of colonies already established in laboratory for many generations. © 2011 The Society for Vector Ecology.
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            Sampling methods for phlebotomine sandflies.

            A review is presented of methods for sampling phlebotomine sandflies (Diptera: Psychodidae). Among approximately 500 species of Phlebotominae so far described, mostly in the New World genus Lutzomyia and the Old World genus Phlebotomus, about 10% are known vectors of Leishmania parasites or other pathogens. Despite being small and fragile, sandflies have a wide geographical range with species occupying a considerable diversity of ecotopes and habitats, from deserts to humid forests, so that suitable methods for collecting them are influenced by environmental conditions where they are sought. Because immature phlebotomines occupy obscure terrestrial habitats, it is difficult to find their breeding sites. Therefore, most trapping methods and sampling procedures focus on sandfly adults, whether resting or active. The diurnal resting sites of adult sandflies include tree holes, buttress roots, rock crevices, houses, animal shelters and burrows, from which they may be aspirated directly or trapped after being disturbed. Sandflies can be collected during their periods of activity by interception traps, or by using attractants such as bait animals, CO2 or light. The method of trapping used should: (a) be suited to the habitat and area to be surveyed, (b) take into account the segment of the sandfly population to be sampled (species, sex and reproduction condition) and (c) yield specimens of appropriate condition for the study objectives (e.g. identification of species present, population genetics or vector implication). Methods for preservation and transportation of sandflies to the laboratory also depend on the objectives of a particular study and are described accordingly.
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              Seasonal Dynamics of Phlebotomine Sand Fly Species Proven Vectors of Mediterranean Leishmaniasis Caused by Leishmania infantum

              Background The recent geographical expansion of phlebotomine vectors of Leishmania infantum in the Mediterranean subregion has been attributed to ongoing climate changes. At these latitudes, the activity of sand flies is typically seasonal; because seasonal phenomena are also sensitive to general variations in climate, current phenological data sets can provide a baseline for continuing investigations on sand fly population dynamics that may impact on future scenarios of leishmaniasis transmission. With this aim, in 2011–2013 a consortium of partners from eight Mediterranean countries carried out entomological investigations in sites where L. infantum transmission was recently reported. Methods/Principal Findings A common protocol for sand fly collection included monthly captures by CDC light traps, complemented by sticky traps in most of the sites. Collections were replicated for more than one season in order to reduce the effects of local weather events. In each site, the trapping effort was left unchanged throughout the survey to legitimate inter-seasonal comparisons. Data from 99,000 collected specimens were analyzed, resulting in the description of seasonal dynamics of 56,000 sand flies belonging to L. infantum vector species throughout a wide geographical area, namely P. perniciosus (Portugal, Spain and Italy), P. ariasi (France), P. neglectus (Greece), P. tobbi (Cyprus and Turkey), P. balcanicus and P. kandelakii (Georgia). Time of sand fly appearance/disappearance in collections differed between sites, and seasonal densities showed variations in each site. Significant correlations were found between latitude/mean annual temperature of sites and i) the first month of sand fly appearance, that ranged from early April to the first half of June; ii) the type of density trend, varying from a single peak in July/August to multiple peaks increasing in magnitude from May through September. A 3-modal trend, recorded for P. tobbi in Cyprus, represents a novel finding for a L. infantum vector. Adults ended the activity starting from mid September through November, without significant correlation with latitude/mean annual temperature of sites. The period of potential exposure to L.infantum in the Mediterranean subregion, as inferred by adult densities calculated from 3 years, 37 sites and 6 competent vector species, was associated to a regular bell-shaped density curve having a wide peak center encompassing the July-September period, and falling between early May to late October for more than 99% of values. Apparently no risk for leishmaniasis transmission took place from December through March in the years considered. We found a common pattern of nocturnal females activity, whose density peaked between 11 pm and 2 am. Conclusions Despite annual variations, multiple collections performed over consecutive years provided homogeneous patterns of the potential behavior of leishmaniasis vectors in selected sites, which we propose may represent sentinel areas for future monitoring. In the investigated years, higher potential risk for L. infantum transmission in the Mediterranean was identified in the June-October period (97% relative vector density), however such risk was not equally distributed throughout the region, since density waves of adults occurred earlier and were more frequent in southern territories.
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                Author and article information

                Journal
                Parasite
                Parasite
                parasite
                Parasite
                EDP Sciences
                1252-607X
                1776-1042
                2017
                15 November 2017
                : 24
                : ( publisher-idID: parasite/2017/01 )
                Affiliations
                [1 ] Monte L. Bean Life Science Museum, Brigham Young University, 2103 MLBM, Provo, UT 84602 USA
                [2 ] 2 Place du Temple, 30440 Sumène France
                [3 ] Division of Entomology, Walter Reed Army Institute of Research, 503 Robert Grant Ave., Silver Spring, MD 84910 USA
                [4 ] Department of Parasitology, Faculty of Sciences, Charles University in Prague, Vinicna 7, 128 44, Praha Czech Republic
                Author notes
                [* ]Corresponding author: plawyer349@ 123456verizon.net
                Article
                parasite170051 10.1051/parasite/2017041
                10.1051/parasite/2017041
                5687099
                29139377
                © P. Lawyer et al., published by EDP Sciences, 2017

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

                Page count
                Figures: 21, Tables: 3, Equations: 0, References: 67, Pages: 39
                Categories
                Special Issue - ISOPS 9 - International Symposium on Phlebotomine Sandflies
                Research Article

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