Training follow-up & evaluation External Quality Assessment and MosPictoQuizz WP4 Medical entomology - December, 2017
REVIEW Novembre 2017
Reminder of the capacity building activities
Training needs assessement • Questionnaire • Tailored trainings
2014
2015
Training courses • Capacity building in mosquito vector of arboviruses • In Serbia, Turkey, Tunisia • For the 19 participating countries
2016
Training follow-up & evaluation • EQA (External Quality Assessment) • MosPictoQuizz
2017
2018
EQA (External Quality Assessement) on mosquito identification
To evaluate the laboratory capacity to correctly identify vectors
Focuses on the mosquitoes (adult and larva) For all the medical entomology laboratories of the MediLabSecure network.
Participant laboratories 19 laboratories from the medical entomology network (WP4) Kosovo / Institute of Vet. medecine Fac of Univ. of Prishtina, Fac of Agri and Vet.
Bosnia &Herz. / Lab. Diagnostic Veterinary Institute of the Rep. of Srpska
Serbia / Lab for med and vet entomology Fac of Agriculture, Univ. Novi Sad
Montenegro / Lab for applied zoology Biotechnical faculty
Moldova / Lab of systematics and molecular phylogeny Institute of zoology Ukraine / Lab of Especially Dangerous Infections Epizootology State Body "Ukrainian I.I. Mechnikov Research AntiPlague Institute of Ministry of Health of Ukraine" Turkey / HU-ESRL-VERG (Vector ecology research group) Hacettepe Univ.
FYROM / Lab for virology and molecular diag Institue of Public Health
Georgia / The R. Lugar Center for Public Health Research National Center for Disease Control and Public Health
Albania / Entomology lab Institute of Public Health
Armenia/ Reference lab center National Center for Disease Control
Morocco / Medical entomology lab Institut National d'Hygiène Morocco / Lab. des Maladies Vectorielles Institut Pasteur
Lebanon / Lab of immunology National Center for Disease Control and Public Health
Algeria / Eco-epidémiologie parasitaire et génétique des populations Institut Pasteur
Jordan / Parasitic and Zoonotic Diseases Department Ministry of Health
Palestine / Vector Control Unit Ministry of Health Tunisia/ Medical entomology lab Institut Pasteur
Libya / Lab of parasitology and vector born diseases National Center for Disease Control
Materials and methods A mosquito box composed of: - 7 or 8 mounted adult mosquitoes - 4 mosquito larvae in alcohol All the specimens were fresh, collected the previous few months.
All available resources were accepted to support identification (dichotomical keys, interactive key for mosquito species, molecular methods‌).
Constraints Despite the precautions regarding the provided materials, some specimens could have been damaged during the transportation. This option has been considered and it has been admitted that the challenging determination of poor preserved specimen relies within the area of the medical entomology expertise. Participants have been invited to take this challenge and to manage this possibility the best they could.
9/143 specimens were considered as damaged by the participating labs. Damaged specimens were not taken into account in the results.
Methods and resources used by participants Resources used for identification 8
100 % morphological identification
7 6 5 4 3
2 1 0 MosKeyTool only
Dicho key + MosKeyTool
Dicho key only
Dicho key + Books (Becker, Harbach)
All labs performed the exercise by
morphological identification, using interactive identification key and dichotomical key.
Dichoto key + MosKeyTool + Books (Becker, Harbach)
Results General results of the mosquito identification from the participant laboratories (rate of correct answers)
100%
87%
90% 80%
80%
77%
% right answers
70% 60%
66% 62%
65% Genus
50%
Species 40% 30% 20% 10%
0% Adults
Larvae
All (adults+larvae)
Results per lab (adults + larvae) General results of the mosquito identification (adults + larvae) from the participant laboratories 100% 90%
% right answers
80% 70% 60% 50%
40% 30% 20% 10% 0%
Lab1 100%
Lab2 100%
Lab3 100%
Lab7 100%
Lab4 92%
Lab5 92%
Lab6 92%
Lab8 100%
Lab9 83%
Species 100%
100%
100%
92%
92%
92%
92%
83%
75%
Genus
Total rate of correct answers for all labs:
Lab10 Lab11 Lab12 Lab13 Lab14 Lab15 Lab16 Lab17 Lab18 Lab19 92% 75% 83% 73% 82% 64% 67% 45% 44% 42% 64%
63%
58%
50%
45%
45%
33%
18%
Genus identification: 80% Species identification: 65%
11%
8%
Results per lab (adults) General results of the mosquito identification (adults) from the participant laboratories 100% 90%
% right answers
80% 70% 60% 50% 40% 30% 20% 10% 0%
Lab1 100%
Lab2 100%
Lab7 100%
Lab3 100%
Lab8 100%
Lab6 100%
Lab4 88%
Lab5 88%
Lab9 75%
Species 100%
100%
100%
100%
100%
100%
88%
88%
75%
Genus
Total rate of correct answers for all labs:
Lab12 Lab10 Lab11 Lab16 Lab13 Lab14 Lab15 Lab17 Lab18 Lab19 88% 86% 50% 75% 57% 71% 43% 43% 40% 38% 63%
57%
50%
50%
43%
43%
43%
29%
Genus identification: 77% Species identification: 66%
20%
0%
Results per lab (larvae) General results of the mosquito identification (larvae) from the participant laboratories 100% 90%
% right answers
80%
70% 60% 50% 40% 30% 20% 10% 0%
Lab1 100%
Lab2 100%
Lab4 100%
Lab5 100%
Lab3 100%
Lab7 100%
Lab9 Lab10 Lab13 Lab11 100% 100% 100% 100%
Lab6 75%
Lab8 Lab14 Lab15 Lab12 Lab18 Lab19 Lab16 Lab17 100% 100% 100% 75% 50% 50% 50% 50%
Species 100%
100%
100%
100%
100%
75%
75%
75%
50%
Genus
75%
75%
Total rate of correct answers for all labs:
75%
50%
50%
50%
50%
25%
Genus identification: 87% Species identification: 62%
0%
0%
List of species to identify Adult species
Larva species
List of adult species to identify for the EQA Species (adult) Aedes albopictus Aedes vittatus Culiseta longiareolata Orthopodomyia pulcripalpis Aedes vexans Culex pipiens Culex hortensis Anopheles maculipennis s.l. ≠Culicidae Aedes caspius Culiseta annulata Anopheles claviger s.l. Total
Nb specimens to identify 19 19 17 19 15 18 11 6 4 3 2 1 134
Nb of adult specimens to identify per lab: 7
or 8
List of larva species to identify for the EQA Species
Nb specimens to identify
Aedes vittatus Aedes detritus Culiseta longiareolata Orthopodomyia pulcripalpis Total
Nb of larva specimens to identify per lab: 4
19 19 19 19 76
Results per species (adults) General results per species of the adult mosquito identification from the participant laboratories 100% 100%
95%
90%
82%
80%
74%
71%
71%
74% 68%
67%
70%
% right answers
73% 63%
67%
56%
60%
45%
50%
Genus Species
40% 30% 20% 10% 0% Aedes albopictus
Culiseta longiareolata
Aedes vittatus
Aedes vexans
Total rate of correct answers for all labs:
Orthopodomyia pulcripalpis
Culex pipiens
Culex hortensis
Genus identification: 77% Species identification: 66%
Results per species (larvae) General results per species of the larva mosquito identification from the participant laboratories 95%
100%
90%
84%
84%
74%
80%
% right answers
84%
84%
70% 60%
47%
Genus
50%
37%
40% 30% 20% 10% 0%
Orthopodomyia pulcripalpis
Culiseta longiareolata
Total rate of correct answers for all labs:
Aedes detritus
Aedes vittatus
Genus identification: 87% Species identification: 62%
Species
Results Are the EQA results linked to the presence/absence in the country ?
% of species right answers 100% 90% 80% 70%
68%
57%
60% 50% 40% 30% 20% 10% 0%
Species present in the country
Species absent in the country
EQA feedback For your lab, this testing has been...
Success rate EQA vs estimated difficulty level 120% Lab1
Difficult 23%
Easy 59%
Lab2
100%
Lab3
% right answers
Very difficult 6% Very easy 12%
Lab4
80%
Lab5 Lab6
60%
Lab7 Lab9
40%
Lab10
Lab11 20%
Lab12 Lab13 Lab14
0% Very easy
Easy
Difficult
Very difficult *5 non-respondent labs
EQA feedback Number of person per lab who performed the EQA
8 7 6 5 4 3 2 1 0 1
2
3 or more
Don't know
EQA comments… … about the damaged materiels Some samples were broken/damaged
Some insects have been damaged, which could lead to errors in identification
… about the EQA benefits
Thank so much for this opportunity to evaluate our ability to identify mosquito
… about the difficulty to identify adult or larva
For our lab in general it will be difficult
We had a bit difficulties during the larvae identification!
This is a good exercise for our laboratory. We found difficulties in identifying adults.
Thank you
Overview of the species to identify
Aedes albopictus An invasive species Mainly black with white stripes on legs and dorsal abdomen. silvery median white line on the scutum
Aedes vittatus Scutum with 6 pale spots, tibiae with median pale ring are diagnostic characters.
Aedes vittatus The larva is recognisable by the implantation of the seta 1-S between the last and the penultimate tooth of the siphon comb.
7/17 labs confused this species with other Aedes sp.: Aedes aegypti (3), Aedes vexans (2), Aedes albopictus (1), Aedes cinereus or rossicus (1)
Aedes vexans Tarsi with pale basal rings and tergum with bilobed basal bands.
Culiseta longiareolata Easy to identify due to the presence of longitudinal lines on the scutum and rows of pale patches on the femur.
3/17 labs confused this species with Orthopodomyia pulcripalpis
Culex pipiens It's impossible to differentiate Culex pipiens s.s. from Culex quinquefasciatus. Difficult to differentiate from Culex torrentium, male genitalia examination is necessary. Confirmation with molecular techniques is recommended.
The apex of all femora with narrow spot of yellowish scales. Tarsi are uniformly dark brown.
Culex hortensis This species has numerous white scales on the thoracic pleurae. And apical pale bands that extends forward in a median triangle on the abdomen.
Aedes detritus Impossible to differentiate from Ae. coluzzi with only morphological criteria. Isoenzymes are diagnostic and a multiplex PCR will be useful for identification. Differentiable also by bioecological specificities.
7/17 labs confused this species with other Aedes sp.: Aedes vexans (3), Aedes vittatus (2), Aedes caspius (1), Aedes rossicus (1)
Orthopodomyia pulcripalpis Distinctive pattern on the scutum and white scale patch on the basal part of the wing allows to identify this species very easily.
5/19 labs confused this species with Aedes aegypti
Orthopodomyia pulcripalpis Abdominal plates on segments VI, VII, and VIII are diagnostic for the genus Orthopodomyia.
MosPictoQuizz
To maintain the mosquitoes species determination among the routine activities A bimonthly online picture activity to identify adult mosquitoes
Materials and methods A online picture quiz open to all, started in January 2017. Every 2 months, a set of pictures of an unidentified mosquito are proposed for identification.
The results are analyzed and reported to participants the following month, accompanied by a mosquito factsheet focusing on the characteristics of the given species, its geographical distribution, the species not to be confused with, etc.
Mosquito factsheet
MosPictoQuizz#1 results
98%
Aedes albopictus
gave the right answers for the genus identification
91% gave the right answers for the species identification
46 participants
19 countries
Aedes albopictus is currently the most invasive mosquito in the world. The contrast between dark and silver scales, and the median line on the scutum are indicative. Note the scutellum in three parts, covered of silvery scales.
MosPictoQuizz#2 results
97%
Anopheles sacharovi
gave the right answers for the genus identification
80% gave the right answers for the species complex identification Anopheles sacharovi is an important malaria vector of the Maculipennis complex. The wing pattern of Anopheles maculipennis s.l. -absence of a patch of clear scales on the wing fringe- is here typical.
33 participants
14 countries
MosPictoQuizz#3 results
90%
Culiseta annulata
gave the right answers for the genus identification
90% gave the right answers for the species identification
29 participants
17 countries
Culiseta annulata is a widespread and common mosquito in Western Europe and parts of the Middle East. Diagnostic characters were: well marked clear band on the abdominal terga, spots of dark scales on the wings, etc.
MosPictoQuizz#4 results
100%
Aedes aegypti
gave the right answers for the genus identification
100% gave the right answers for the species identification
26 participants
15 countries
Aedes aegypti is an invasive mosquito recently reestablished in Madeira and around the Black Sea. Diagnostic characters are mainly on the scutum with the famous "lyre-shaped" with the two fine central longitudinal bands (the strings of the lyre!).
MosPictoQuizz#5 results
100%
Culex pipiens
gave the right answers for the genus identification
90% gave the right answers for the species identification
20 participants
13 countries
Culex pipiens s.s., as well as its tropical vicarient Culex quinquefasciatus, are members of the Culex pipiens complex. Morphology of females does not permit to distinguish the two species.
MosPictoQuizz#6 results
100%
Anopheles claviger
gave the right answers for the genus identification
57% gave the right answers for the species identification Anopheles claviger s.s., as well as its sibling species Anopheles petragnani, are members of the Claviger Complex. An. claviger s.s. occurs across most of Europe, up to the Middle East and North Africa, whereas An. petragnani is so far restricted to the western Mediterranean region.
35 participants
13 countries
MosPictoQuizz participation Nb of participant /
Nb of country /
50 46 45 40 35 35
33 29
30
26 25 20
19
20
17 15
14
15
13
13
10
5 0 #1
#2
#3
#4
#5
#6
MosPictoQuizz feedback MosPictoQuizz#1
MosPictoQuizz#3
Summary of results
~50% of the labs (9/19) were able to identify more than 75% of the specimens, which means between 0 and 3 mistakes.
19 participant labs to the EQA
12 different species
210 specimens to identify
Good results (87%)
#6
on genus identification of larvae.
MosPictoQuizz
All the entomological labs are able to identify
invasive species (Aedes albopictus and Aedes aegypti).
Recommendations Mosquito identification is an important step in entomological surveillance. A regular practice, on top of routine activities, is highly recommended. Even though we usually feel more comfortable with larva and/or adult identification, do not neglect any of those two. Practice equally larva and adult identification. The profession of medical entomologist have changed considerably these last decades. Fieldwork remains fundamental as the transmission of infectious agents is above all a question of ecology.
This MediLabSecure activities has been organized by the French National Research Institute For Sustainable Development (IRD). This medical and veterinary entomology group is coordinated by Dr. Vincent Robert and Marie Picard at MIVEGEC unit, IRD.
The MediLabSecure project is supported by the European Commission (DEVCO: IFS/201010/23/_194)