A guide for collecting, preserving and handling insects

[Patchara Sriwichai]

A guide for collecting preserving and handling insects (1 hr) – 5 scores

If you take the function to be the expert of medical entomologist, what is your plan and actions to support and react for the situation below?
1. Routine work of vector surveillance for malaria  (2.5 scores)
2. There is the outbreak of malaria in Thai-Myanmar border in Tak province.  (2.5 scores)
Please explain the detail in term of

– Objectives

– Planning

– Program activities

– Methods

– Analysis

– Reports

[Anonymous]

I’m Oranit Deesitthivech ID:6436419

If you take the function to be the expert of medical entomologist, what is your plan and actions to support and react for the situation below?
1. Routine work of vector surveillance for malaria
Malaria is spread to people through the bites of infected female Anopheles mosquitoes
which is an acute febrile illness caused by Plasmodium parasites. There are 5 parasite species that cause malaria in humans including Plasmodium falciparum, Plasmodium malariae, Plasmodium vivax, Plasmodium ovale, Plasmodium knowlesi.
– Objectives
what, where, when(how long), why, do you want to do? Because of all of these must have under money to manage. You might to think the important, appropriate and worthiness that want to do.
– Planning
-What do you do in the field and laboratory.
-How to get the data
-How to analysis
-How to conclusion and interpretation the result
-How to monitoring and control in the long term

2. There is the outbreak of malaria in Thai-Myanmar border in Tak province.
Please explain the detail in term of

– Objectives
*To track malaria vector densities (for selection and timing of vector control malaria program)
*To investigation effective and responsive to any threats to a successful control and elimination program.
– Planning (Under funding)
*Where is you want to surveillance? (You must to think about that area have problem malaria such as case report, Resting place and breeding place of vector, transportation of people)
*What time you want to go?
*What do you do?
*How do you do?
*How do you get the data?
All of these you must to think and good planning before go to the real situation

– Program activities
*To collecting mosquitoes for outdoor and indoor (what’s time, what’s stage mosquitoes of do you want for you to the appropriately method)

– Methods
*Preparing instrument/equipment for various techniques
*Collection techniques for adult
Biting collection: – baited catches (landing catch, animal-baited net)
Resting collection:- indoor resting (pyrethrum spray sheet, aspirator)
– outdoor resting: direct (aspirator, sweeping net), indirect (artificial shelter)
Trapping collection: – light trap, sound trap, gravid trap
*Collection techniques for immature collections
Dipping: dippers
Netting: hand nets
Pipetting: droppers
Siphoning: rubber tubes
Special techniques: Uproot, cylinder (Mansonia)
*Record the data including date, place(coordinates), time, sample, etc.
*If you want preserving and handling mosquitoes for study in the future about morphology you should do the appropriately method for killing and preserving. Killing method by Cold at 4 ºC or-20 ºC or Killing Jars/Bottles. Preserving method by Ethanol (grain or ethyl alcohol) mixed with water (70 to 80 percent alcohol) is usually the best general killing and preserving agent.
*How to control/stop the outbreak?
– Education to villagers/traveler
– Protection from mosquitos bite (Wear long-sleeved clothing and long pants, Use a mosquito net, Spray
an insecticide or repellent)
– Eliminate breeding places

– Analysis
When you get the raw data you can bring to analysis or record with Microsoft for generate the data what do you want.

– Reports
The last step can conclusion and interpretation the result so led to what is the next step in the present or the future (what do you have to do).

[Patchara Sriwichai]

Dear Oranit,
Good start for our discussion.
in Routine works; please more explain in details what is the objectives and plans based on situation. Do not mention only “what, where, when(how long), why, do you want to do? Because of all of these must have under money to manage. You might to think the important, appropriate and worthiness that want to do.
– Planning
-What do you do in the field and laboratory.
-How to get the data
-How to analysis
-How to conclusion and interpretation the result
-How to monitoring and control in the long term”
Please give the schedule plan exactly what where when and how frequency you plan to take the action.

for #2 again, please explain in detail what activities will be done.
what method you will select. don’t mention over all as I gave in slide. You have to make decision why you select this period or method and how to analyze data and records?
### please do not all detail in slide to answer but try to analyses and criticize what the best one you select? and why?

See you can feel free to discuss……..

[VANHEUANG PHOMMADEECHACK]

Hello everyone, I am Mr. Vanheuang Phommadeechack Student ID: 6436660

This is an interesting topic for me in terms of medical entomology and epidemiology. let’s see my opinion below:
1. For routine work in vector surveillance of malaria, i think the objective to conduct this task is for planning for vector control program and for early warning of disease outbreaks in the specific area and for evaluating the effect of vector control.
– In my opinion, we should start with planning first to design the study and select the appropriate method for sampling like cluster sampling, systematic sampling, random sampling.
– After that, we will plan for the collection technique and the form such as data collection form, data record form, plan to record data in “Excel sheet”, then plan for the equipment to use for sample collection, and plan for mapping (where to collect the sample), set the schedule and timeline to collect the sample.
– Choose the suitable type of collection method like adult collection by using resting collection, biting collection and trapping collection.
– Record and preserve the sample in appropriate way for further investigation which based on the purpose of study then transport the sample to the laboratory, for example keep in cryotube or Eppendorf tube with cilica bead.
– Then we will select type of vector survey, for example we can use preliminary survey for short term, regular or trend observation for monthly or based on season, and spot check. This can be used to evaluate the effect of vector control.
– Analysis of the data from lab results and report the result of the surveillance including the description of the study, worksheet report, discussion, comment, interpretation and conclusion.

2. For the situation of malaria outbreak in Thai-Myanmar border in Tak province, If we would like to take part for the identification of disease outbreak or vector surveillance, I think we should proceed as following process.
– Set the objective that can answer or can solved the problem. For the objective of vector surveillance in Thai-Myanmar border in Tak province is to identify the cause of disease outbreak, for species identification of mosquitos that caused the disease outbreak in that area, for vector controlling, for early warning of malaria outbreak in that area and to evaluate of vector control.
– Planning:
For the study design I think Cross-sectional study because this is the most appropriate to collect the sample in the specific period for vector surveillance. To collect the sample, I choose the systematic sampling method to collect the mosquito or collect the sample nearing malaria case report areas to get representative of sample in that area, writing study protocol calculate the sample size.
– Program activities: setting the timeline and schedule to collect the sample, listing the activities to be done in every day
– Methods:
. Equipment and tools: preparing the equipment that will be use in sample collection and preparing the data colleting form.

. Vector collecting technique: I will use trapping method to collect the adult anopheles mosquito such as light trapping, BG trapping and set the trap in early evening and leave it overnight with trap labeling and mapping record, then collect the trap in the morning of next following day.
. Sample preservation: sample will preserve in silica bead with labeling or sample coding and record in excel sheet.
. Sample transportation: sample will pack in a primary and second container with icepack and sealed before transport to the laboratory
. Laboratory investigation: Morphology, molecular identification for species (COI gene) and for malaria detection in mosquito (qPCR for malaria detection).
– Analysis: Data of sample and laboratory test results will be recorded in the excel sheet, then will summarize the data based on identified species of morphology and molecular, and analyze the data according to molecular laboratory results of malaria detection in mosquitoes using qPCR.
– Reports: the result of the study will be reported by data summary with the conclusion of the result, the description of the study, discussion, comment, interpretation of the data analysis and conclusion of the study.

Happy discuss and hear comments and feedback from Ajan and friends.

[Anonymous]

I’m Oranit Deesitthivech ID:6436419

1. Routine work of vector surveillance for malaria
The objective of vector surveillance for malaria to surveillance, monitoring and planning control program for evaluate early warning outbreaks of malaria disease and public health problem that area
First, start with planning of work
-What do you do in the field and laboratory: Design the study and select the appropriate design study. Design method for collect samples techniques for adult or/and larva.
-How to get the data: Design the record paper for note the detail of a sample (can tack the data after)
-How to analysis: After get the raw data put it in the excel to easy analysis and look the data
-How to conclusion and interpretation the result: Make the graph help to conclusion that it’s simple to see the trend of data so I can conclude and can know in the next step how can I do for monitoring and controlling.

2. There is the outbreak of malaria in Thai-Myanmar border in Tak province.
Please explain the detail in term of
– Objectives
*To track malaria vector densities (for selection and timing of vector control malaria program)
*To investigation effective and responsive to any threats to a successful control and elimination program.
– Planning (In the detail I think it’s similar in the first question)
*Where is you want to surveillance? (You must to think about that area have problem malaria such as case report, resting place and breeding place of vector, transportation of people)
*What time you want to go?
*What do you do?
*How do you do?
*How do you get the data?
All of these you must to think and good planning before go to the real situation

– For methods I think I will collect samples adult and surveillance larva to see the real problem it’s form where if it’s form of bleeding site maybe easy to control and eliminate.
– Collection techniques for adult I use trapping collection by BG trapping it’s easy to use, low chance bitted by mosquitoes. For larva surveillance only and record the data (where, number of larva) if found that I will suggest to villager or villager head about how do you do for eliminate larva (Both method I will record the data including date, place(coordinates), time, sample, etc.
*And then if I found the sample complete morphology or new thing I will preserving and handling mosquitoes for study in the future about morphology you should do the appropriately method for killing and preserving. Killing method by killing Jars/Bottles and Preserving method by Ethanol (grain or ethyl alcohol) mixed with water (70 to 80 percent alcohol) because the best general killing and preserving agent.
After that get the data put it in the excel to easy analysis or with Microsoft for generate the data. The last step can conclusion and interpretation the result so led to what is planning in the present or the future.
So, I can see the problem and can tell or suggest to villagers How to control/stop the outbreak? By provide education to villagers/traveler, how to protection from mosquitos bite (Wear long-sleeved clothing and long pants, Use a mosquito net, Spray an insecticide or repellent), how to eliminate breeding places.

Thank you for suggest me 🙂

[RATTANALAK JITTUNGDEE]

Rattanalak Jittungdee 6438165

1.Routine work of vector surveillance for malaria
I agree with Mr.Vanheuang’s opinion in Item 1 that surveys should be designed to collect data, for example, if we want to collect data on adult mosquitoes or larvae. We should choose that Anopheles spp. that are the main vectors for malaria. The life period and habitat of this species should be studied. To store information according to the data recording plan that has been set up. Based on the objective of the survey is to monitor malaria vectors.
– Long-term sampling is a survey that is often done to look at the increase or decrease of vectors in that area. Although it is a longer method to draw conclusions about how the disease fluctuates.
– Short-term sampling is a survey that often goes into areas where the infection has been reported. This may be surveyed to confirm the prevalence of vector vectors in that area and the rate of vector transmission. The results will lead to an assessment to control the situation in that area.

2. There is the outbreak of malaria in Thai-Myanmar border in Tak province. (2.5 scores) Please explain the detail in term of
– Objective: This part depends on what we want to do for what purpose.
 To identify the source of malaria outbreaks in reported areas
 To control the spread of disease
 To study the spread of disease and its vectors
– Planning: The planning will be adjusted according to the objectives of the work. which we must have knowledge of the habitat and behavior of mosquitoes and choose a study method to suit the results we want
 Identify the carrier mosquito species: We need to know the terrain of the outbreak area. Corresponding to the habitat of what types of mosquitoes we may encounter in the area. When is the time during which mosquito species are active? A cross-sectional study may be used to collect data.
 Outbreak Control: We need to know what the outbreak area looks like. life of villagers How is the habitat How many outbreaks How are the number of vectors and the rate of the outbreak? If the infection is more children or adults, a retrospective study or cross-sectional study may be used.
 Disease transmission rates and vectors: Know how many mosquito breeding areas the area has. What is the number of infected people? The number of mosquitoes that can be found in the survey such as using methods capture-recapture method to measure mosquito population prevalence will affect the spread of the disease as well. A retrospective study or a cross-sectional study may be used.
– Program activities: Determining what to do each day, for example, in the morning to collect samples or data in Area A, and in the afternoon to collect samples or data in Area B, etc.
– Methods: Methods used to collect data or collect samples. which should have a form to record information The equipment used to collect samples corresponds to the survey area. including methods for preserving samples for experimentation or inspection
 The use of BG trapping to trap adult mosquitoes or the use of devices used to catch larvae in water sources that may have Anopheles mosquitoes. Follow different areas of the survey area and map where traps are placed. and save the data
 Preservation of collected samples which we want the example to be complete. for effective monitoring such as Collection in tubes containing silica beads, for example, and labeled with type of sample, date of collection, area of collection. including sample collectors
 Transport of samples that can be stored in a sealed and strong container to prevent damage or loss of samples
 Laboratory Examination: Morphological Classification, Molecular species identification including the detection of malaria in mosquito samples obtained
– Analysis: Bringing the data obtained from local surveys and laboratory results to be recorded in a digital system such as Excel, then analyzed and summarized.
– Reports: describe the results of the survey. Discuss how well the results meet the objectives. result meet the purpose or not What needs to be improved in the next study?

[TIPAWAN AMMARIT]

Hello everyone. I’m Tipawan Ammarit student ID: 6438166

1. Routine work of vector surveillance for malaria.
In this point, I’m disagree with my friends that they said we have to start with planning first. But I’m gonna start with the important part of this research is objectives setting.
– Objectives setting: In my opinion, we have to start with this process be cause we have to determine the limitation and what’s we gonna get from this work. Thus objectives for routine work of malaria vector surveillance are observe the species, density, quantity of vectors and moreover we can determine to collect the vector samples for further study.
– Planning: Next, we must plan everything including the vector species which have to focus on (in this case we focus on malaria vector like mosquitoes genus Anopheles), types of sampling (select the appropriate method for the study area such as random sampling, systematic sampling, and cluster sampling), types of mosquito survey (choose the appropriate one like preliminary surveys for basic and short-term surveys, regular observations for long-term surveys, spot checks for monitoring mosquito populations, and foci investigations for identify the best approaches to control), detailed schedule sampling program for each hour, list of equipments, Data sheet preparing (, list techniques which have to use (biting, resting, trapping, netting for adult collection or use aspirator, strainer for larvae collection or ovitrap, pipette for eggs collection).
– Record and preserving: after that we have to go to the field at the selected and interested areas thus run the schedule like we already planned then record every important data that we got on the prepared data sheet, moreover we’ll bring appropriate tools for Anopheles mosquito collection at several stage like adults, larvae, or eggs. The main thing is that preserving samples for bring back to the laboratory study, we must used the dry ice, silica bead, or RNA later to keep the DNA/RNA/protein maintainly.
– Analysis: afterwards bring the data to analysis process may be use MICROSOFT EXCEL and produce the graft or table for analyse and interpret easily. Finally, results must present to the other people, thus we must have communication skill to lead them understand our work easily and clearly.

2. There is the outbreak of malaria in Thai-Myanmar border in Tak province.
– Objectives:

1. 1) Found out the mosquito species which is main vector for malaria disease to human that cause outbreak in Thai-Myanmar border, Tak province.
2. 2) Found out the quantity and density of vector mosquitoes in outbreak areas.
3. 3) Found out the control vector method for reduce spreading rate of malaria disease.

– Planning: We must start with collect related informations from the hospitals or health care centers for evaluate the situation of this outbreak (e.g. mapping, case reports). The most important we have to concern about harmful and danger of malaria disease in this area, we must prepare ourselves and tools before go to investigate in that high risk area. Next, we have to select appropriate sampling method match with study areas (in this case we can use random sampling for unbiased statistical properties), choose the types of mosquito survey (in this case it’s be preliminary surveys cause we have to solve the problem of malaria outbreak hastily for help people in high risk areas), then choose the stage of samples that we gonna get (for species identification we must collect adults, for quantity and density examination I choose to collect adults and larvae) and methodology (select to use light microscope to species identification, use trapping, netting, and dipping for adults and larvae collection), prepare data sheets and sampling program. And I’m agree with Rattanalak that told use technique capture-recapture to evaluate population of Anopheles mosquitoes this method it’s very great I think.
– Program activities: We have to program activities in everyday and per hour because if we don’t prepare, it’ll waste our time uselessly. Hence timetables for everyday are very useful. And for accurately collection and experiment we must select appropriate time period to do each activity for example collect adult stage at late evening (5 pm. to 9.30 pm.) is biting period of Anopheles mosquitoes.
– Method: We use mosquito traps with sweaty coats for induced adult mosquitoes with light and CO2. Then we use net and aspirator for collect adults which flying and resting near the human, moreover use colander (net-like) to dipping and collect larvae at breeding place near human houses daily. Next, we gonna record the data in data sheets completely. And species identification by using microscope at the field and record the number of mosquitoes that we got. then preserving the samples by using individual tube for whole body mosquito collection (maybe for entomological specimen center) or genetic material (DNA or RNA) extraction and stored on ice/dry ice for keep genetic materials to PCR technique in laboratory for high accuracy species identification and find the malaria parasite in the samples by molecular technique. Lastly, we gonna find method to control mosquito population by reduce the contact between mosquitoes and humans by using larvicides or larvae predators, indoor residual spraying and insecticide-treated bed nets thus record the number of case reports at hospitals and health centers.
– Analysis: We bring the collected data to analyze with MICROSOFT EXCEL and produce to be compared graft or table and interpret results to easily understand to other people. Moreover, we have to consider about the limitation of this short-time survey, the results don’t completely done thus interpretation may be change if we study for long-time.
– Report: We have to describe the research introduction, methodology, results (show the reliably results), discussions, and conclusion at summary presentation by easily understand word. Finally, show the trend of further developing research.

Thank you very much.

[PANATCHAYA SOMAREE]

Hello everyone, It’s me Panatchaya Somaree Student ID 6438168.

1. I will begin my discussion with the routine work of vector surveillance for malaria.
First of all, I agree with one of my friend’s point that in order to do vector surveillance, you need to understand the objectives beforehand. The objectives indicate the scope of work and facilitate a better planning.
Objectives: For the objectives of the vector surveillance would be to identify the transmission pattern, to investigate the habits and habitats of the local vector species, to monitor the seasonality and abundance of vector population in term of early warning of malaria outbreak and also to plan for a vector control program (select the appropriate method with appropriate time and place).
Planning: After we understand the objectives, we should consider the timing and the manpower in order to select the appropriate study design (such as cross-sectional study, retrospective study or experimental study) and the appropriate sampling methods (how many samples and which methods).
– The suitable types of mosquito survey (such as preliminary survey, regular/trend observation, spot checks or foci investigation) should be considered and also the collection techniques of larvae or adults are needed as well.
– We need to concern about the biology of what we collect in this case are Anopheles mosquitoes so we could decide the methods of handling and preserving. Then we could prepare a list of equipments needed (such as dippers for collecting the larvae or silica beads for handling ).
– Moreover, we should also plan what to record (such as time/date, place, collector), how to report and analyze the data for example generate the data with Microsoft Excel and analyze in terms of graphs to facilitate the interpretation and conclusion.

2. For the Thai-Myanmar border in Tak province’s malaria outbreak situation, in term of
– Objectives:
1. To identify the causes of malaria outbreak in this area (such as human factors, seasonal period, insecticide resistance, high vector density)
2. To identify the species of mosquito vector responsible for malaria transmission in this area
3. To effectively control of malarial vector
4. To evaluate the efficacy of the vector control program
– Planning:
1. Evaluate the malaria outbreak situation in this area and gather all related information
2. Select the appropriate study design. I agree with my friends that the cross-sectional study should be conduct in this case because it’s quick and we want to collect the data only during this outbreak period in order to quick control of the outbreak
3. Select the appropriate sampling techniques. We need to know the habits and habitats of this vector species first such as the potential breeding sites or the resting places. In my opinion, we should use the systematic sampling. We need to know the sample size and sampling interval.
4. We should know the timing and the manpower beforehand in order to separate the tasks responsibilities.
– Program activities: The manpower should be considered in order to create timetables. We might separate into groups like the one responsible for the mosquito survey could active in the evening and the one responsible for generating or analyzing data could active during daytime. This could help in less time consuming and balancing the workloads.
– Methods:
There are various methods for mosquito collection based on which stage we want to collect. In this case, for the adult stage of Anopheles mosquito collection, the trapping method should be use such as the BG trapping or CDC light trapping. For the immature stage, the dipping method using the dippers should perform. After mosquito collection, we need good methods in order to maintain the best quality prior to the laboratory detection including handling, preserving and transporting. For handling the mosquitoes, we use silica gel to absorb moisture and prevent fungi. The transportation of alive mosquitoes requires the primary and secondary containers. The primary container must be covered with the ice pack to maintain the cold temperature. The preservation of mosquito can perform in 70-80% of ethanol for immature stage. The adult stage can preserved by pinning. All of the samples must be labelled or coded. Then, the identification of mosquito species will be perform in the next step using the morphology identification and molecular techniques.
– Analysis: Gathering the collected data and generate the data in terms of graphs with the programs such as Microsoft Excel. Some data should be calculated according to the formula before concluding. After analyzing concluding should be done.
– Reports: The reports should include the methods and techniques using, conclusion and results of this program. The explanation whether the results could meet the objectives or not. The discussion about the successful and failure from this program. The methods to deal with those mistakes. As well as, how to bring these informations to apply in the new vector control program.

Thank you very much.

[RATTANALAK JITTUNGDEE]

Dear Ajan Patchara,

These are conclusions regarding the plan and actions to support and react to the case scenario from five of us (Vanheuang, Rattanalak, Tipawan, Oranit, and Panachaya).

1. For routine work in vector surveillance of malaria, the objective is for planning the surveillance of vectors, monitoring, and planning control programs for early warning of disease outbreaks in a specific area, and for evaluating the effect of vector control.
The plan for routine work in vector surveillance includes objective, planning, method, analysis, conclusion, and interpretation of the result.
• Objective setting: Set the objectives of the vector surveillance based on the study such as identifying the transmission pattern, investigating the habits and habitats of the local vector species, to monitor the seasonality and abundance of the vector population in terms of early warning of malaria outbreak and also to plan for a vector control program (select the appropriate method with appropriate time and place).
• Planning: After we understand the objectives, we should consider the study design. Such as type of study, design method, and techniques that will use for collecting the samples and laboratory investigation.
• Method:
– Data collecting: preparing for data collecting and data recording such as the form that will use to record and preserve the sample inappropriate way for further investigation based on the purpose of the study
– Analysis: analysis of the data from lab results using Microsoft Excel and analyze in terms of graphs and tables. Then, conclude the result and report the result of the surveillance by using the description of the study, worksheet report, discussion, comment, interpretation, and conclusion.

2. For the scenario of Malaria outbreak on the Thai-Myanmar border in Tak province,
• Objectives:
– To identify the causes of malaria infection on the Thai-Myanmar border in Tak province
– To identify the species of mosquito vector responsible for malaria transmission in this area
– To eliminate or control malarial vectors on the Thai-Myanmar border in Tak province
– To evaluate the efficacy of the vector control program
• Planning:
– Select the appropriate study design: A cross-sectional study identifying the prevalence of malaria in vectors and patients in this area.
– Select the appropriate sampling techniques: systematic sampling method to collect the sample to get representative of sample in that area and we need to know the sample size and sampling interval.
– Select the appropriate technique for laboratory investigation
– Planning for the timeline of the project to set a schedule for sample collecting, laboratory investigation, and data analysis
• Program activities: In Determining what to do each day, we should know the timing and the manpower beforehand to separate the tasks responsibilities.
• Methods: Methods used to collect data or collect samples. which should have a form to record information The equipment used to collect samples corresponds to the survey area including methods for preserving samples for experimentation or inspection and technique that will use in laboratory investigation.
In this case, for the adult stage of Anopheles mosquito collection, the trapping method should be used such as the BG trapping or light trapping. After mosquito collection, we preserve the sample inappropriate ways such as preserving it in silica bead, putting it in the primary and secondary containers with ice-pack, and labeling. Then transport to the laboratory for study purposes. After that, the identification of mosquito species will be performed in the next step using morphology identification and molecular techniques.
• Analysis: Gathering the collected data and generating the data in terms of graphs with the programs such as Microsoft Excel. Some data should be calculated according to the formula before concluding. After analyzing concluding should be done.
• Reports: Reports the investigation result which includes the methods and techniques used, and states the conclusion and results of this program. Discuss the results whether meet the objectives or not. Then, discuss the success and failure or advantages and limitations of this program. The methods to deal with those mistakes. As well as, how to bring this information to apply in the new vector control program. Finally, show the trend of further developing research.
So, we can see the problem and suggest to villagers to know and waring about the disease outbreak in this area and suggest how to prevent and control the malaria outbreak in the Thai-Myanmar border of Tak province.

Thank you very much.

Yours sincerely,
Vanheuang, Rattanalak, Tipawan, Oranit and Panachaya

[Author]

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