Questions related to Mosquitoes
I'm currently investigating the larvicidal efficacy of plant extracts and some pure isolated compounds. Due to limited amount of materials, I may need to use lower volume of water compared to the WHO guidelines (100 - 200 mL). I've seen research papers using 6- and 24-well plates with 1 mL of water per well. Is there a huge difference to the outcomes between 1 mL and 100 mL of water being used? Thank you in advance!
Who can tell me is there a link between the knock down time 50 (KdT50) and the resistance mechanism of the mosquito? in other words can we say that if the knock down time is high then the resistance is linked to the kdr gene ?
I need to add taxonomic related vouchers for both adult male and adult female tea mosquito bugs in my article. Hence guide me to search for the same.
Can anyone recommend a vendor that sells mosquito salivary recombinant proteins Aed a 1, Aed 2, Aed 3, or Aed a 4, for use in epitope mapping and other immunological assays?
I am doing whole cell patch clamp recordings on mosquito brain cells where I can form gigaseal but after breaking in into the cell there is gigaseal formation again even if I try to break in using suction or zap and the resistance value is usually on the higher side and even if it drops after applying suction the cells are lost then.
I am currently pursuing my research on the dengue-causing mosquito at its various stages. I'm studying the effect of microbial secondary metabolite-based nanoparticles against the vector that carries dengue. In order to extend my study, I would like to use the DENV serotypes to validate the cytotoxicity of the particles that I developed.
Is it possible to share data on mosquito species diversity among countries with land boundaries? The expected result is a revision to the mosquito species checklist between corresponding countries. The idea is that mosquito distribution does not respect the boundaries of different countries. They only respect the suitability of the environment they live in. The diversity data acquisition is based on a study of ecosystems and land use between countries. The endemicity status of each species is also becoming an important consideration. What needs to be done if the species data acquisition is possible?
Dear Scientists (Vector Biologist/ Public Health Entomologist),
Please look into the attached pictures and suggest what could be the reason for such type of mosquito larval mortality where they were covered in a slimy layer and died.
Also kindly suggest if any of you faced same problem and found the solution to avoid this.
I have acess to some rare field samples, that have been stored in absolute ethanol. I know RNA can be extracted from such samples, provided the storage temperature was low (e.g.,
Semi-polar limonoids wanted to be incorporated in O/W nano-emulsion for mosquito larvicidal application. The compound is poorly soluble in the carrier oil, but readily soluble in organic solvents ie ethyl acetate, acetone. The emulsifier I'm using is Termul1284. I have tried mixing the compounds with the emulsifier first, before adding in the oil and water. But I'm having doubts whether this would entrap the compounds in the carrier oil. Any advices? Thank you in advance
I want to assess the infection status of Aedine mosquitoes with regard to dengue virus in Zambia. Very little is known about the prevalence of the disease in the country.
A new microbe was found last year to have the ability to block plasmodium transmission without hurting the mosquitoes, I decided to present a seminar on this... hence my need for a case study/studies to prove my point.
Let say the mosquitoes were collected from different houses for sever days and then want to determine an average density of harvested local mosquito population.
On three separate occasions I have encountered this mite, what seems to be the of same species, attached to the proboscis of Aedes aegypti and Culex erraticus species of mosquitoes. My guess is that it is hitching a ride from something (flower/bird), but I haven't been able to find a mite that matches. I'm hoping an expert here can at least get me to a genus or know what host it is from. Thank you in advance!
Update (4/25/22) - The genus has been determined to most likely be Cheyletiella due to the rounded body shape and large palpal claws but the species is still unknown. Since this post, a couple more mites were found on Culex mosquitoes attached in the same manner (high up on the mosquito's proboscis). Two PCR attempts failed so it is looking like visual ID of the solenidion will be the way to go. As we prepare to begin mosquito trapping again, I am sure more specimens will be obtained for either stacked photography or SEM. Regardless of species, it is looking like this is a phoretic relationship. I have reached out to other mosquito biologists to keep an eye out for these mites while identifying mosquitoes under the microscope.
We need the culture of three mosquito species namely Anopheles stephensi, Aedes aegypti and Culex quinquefasciatus for our insectary establishment. We will be very grateful, if anybody inform us, from where we can get the same.
I am considering the use of CO1 to differentiate between different mosquito populations. Is it an ideal target gene? And does it have better resolution than ND4 and ITS1, ITS2 to differentiate between closely related species?
If I understand correctly, the Sella score (based on Detinova, 1962) assigns unfed female mosquitoes to 1, and gravid to 7. This is in order or the digestion process, with 2 being freshly full-fed.
My question is, what would the Sella score be for a partial fed mosquito be?
I have many individuals that have only taken a little bit of blood (often collected by light traps), but the Sella score has 1 being unfed and 2 being fully fed.
Thank you in advance for any advice, and any references would also be very helpful.
I wonder about the biology of mosquitoes in relation to sex, we can differentiate An. gambiae from An. funestus or Culex pipiens from the larval stage but can we also differentiate their sex during these stages?
I am preparing slides for the genitalia of male mosquitoes of the Culex pipiens complex. I read it is used cellosolve to dehydrate the genitalia. Any alternative to replace it?
I need to hatch a lot simultaneously and they're being very stubborn despite multiple immersions in water.
We are trying to isolate DNA from mosquitoes using chelex and saponin to no available. Anyone with an in-house protocol that works?
We will be grateful.
I am trying to extract RNA from Aedes aegytpi to analyze gene expression in genes associated with the detoxification of insecticides. I am using the SV Total RNA Isolation System from Promega, but I got a very low yield of RNA (from 30 - 50 ng/ul). Can you share any tips or protocols or how have you solve low yield in RNA isolation from mosquitoes. Thank you very much.
can anybody suggest how to extract DNA from insect samples conserved in silica gel (in my case, mosquitoes)? Does anybody know a commericial kit which does that or a particular protocol?
Thanks a lot in advance
I need a method to isolate mosquito cell membrane protein from whole mosquito abdomen. Therefore, I will need to remove the mosquito hemolymph and exoskeleton.
I have not find a clear published method to do that. Any suggestions would be super helpful
Many thanks in advance
Are you aware of research-proven natural remedies that can be used to prevent mosquitoes bites (repellent), either by local application or consuming more amount of such food?
What about after the insect bite: any herbal remedies that is research-proven to be effective in reducing the symptoms?
Hello Researchers. I am conducting a study on the determination of WNV on mosquito samples. I run nested PCR. Bands were shown to almost all samples during carrying out gel electrophoresis. After I sent the PCR product for sequencing service for 5 random samples, and the sequencing result for 4 out of 5 appears to be 100% percentage identification of my targeted virus (WNV). While one sample shows 98% as the highest percentage identification. Hence, I would like to know whether my results are true positive (valid) or false positive? May I know how to determine the false positive result (any criteria or book/journal reference) as my future reference?
My targeted primer is between capsid and pre-membrane.
Thank you for your thought and suggestion.
I need the mosquito eggs to be hatched within a short period of time, but the eggs are taking upto 20 days. What should I do? Please don’t avoid this question.
Hi could anyone share their practical experience about Arboviruses detection from field caught mosquitoes. I have used mosquito samples to perform metagenomic sequencing and passaged on C6/36 cells for Arboviruses detection. I have detected a considerable number of reads for JEV/GETV and the similar samples produced CPE’s on C6/36 cells and also confirmed the particular virus presence by RT-PCR from CPE’s positve C6/36 cells (Japanese encephalitis virus, GETV etc). However when I passaged the same samples on BHK cells, it don’t produce CPE’s.
Thank you in advance.
I am working on mosquito genomics and trying to find a best protocol or method for RNA extraction from a single head and thorax part of an individual mosquito. The RNA will be used for NGS applications. Please suggest.
I downloaded the yeast- CO2 mosquito attractants paper pub. Sept 2017 and found it has a number of typographic errors. The Kline publication (2007) is cited 2 times as #14 and #24. It would be good to fix this and a number of other errors. I cannot figure out how to email Dr. Aziz Muhammed directly through RG or I would do so.
Consider a simplified version of the ODE model presented in Marini et al. 2017 (see section Model calibration of Additional file 1 in that paper).
In this model there are two populations of mosquitoes, that go through their different life stages: egg -> larva -> pupae -> female adult. For simplicity the pupae stage is omitted in this example without loss of information (see attached file 1). When evaluating this system in Wolfram Mathematica and obtaining the Jacobian we arrive at the steady states (see attached file 2).
If we focus on the scenario where all populations are present (i.e. Ec > 0, Ea > 0, Lc > 0, La > 0, Ac > 0, Aa > 0) and substitute the parameters with any values at this steady state, the populations of larvae, Lc and La, exceed their carrying capacities, Kc and Ka respectively.
Why is this happening?
Can populations exceed the carrying capacity if they are only limited by it on a single life stage?
Or is there a problem in the implementation in Wolfram Mathematica?
Malaria caused by Malaria parasites (Plasmodium species) is transmitted by Mosquitoes. Malaria especially in the sub-Saharan African region remains a major public health due to associate high mortality and morbidity particularly in the under-five.
Mosquitoes repellents are a major tool for the control of spread of the disease.
What is your take on the above question?
Hello everybody, I am not a specialist and I want test the effect of some substances on mosquitoes and flies but I have no idea how can I get them and how to keep them alive for treatment?
Thanks in advance
I want to measure mosquito body weight post emergence and would like to find out if I can just take off the wings and weigh the body without drying the mosquito.
Is there any study published and demonstrated which indicates the presence of the agent of rickettsioses? in other arthropods not hematophage.
I'm working with a mechanistic model to predict mosquito abundance with temperature input. I also have a raster with land-use classification for the same area and I now want to know how land-use dictates mosquito abundance (as land-use influences micro-climate). I think I should correct for spatial autocorrelation, but am struggling with how to do that best. One of my ideas is to find out up to what lag distance autocorrelation is present/significant and then do a ANOVA+post-hoc only on cells sampled x lag distance apart.
However, I'm confused about the different approaches amongst different functions/packages in R. On the one hand I tried lm.morantest, which computes Moran's I over the residuals of a linear model (in my case mosquito vs land-use). On the other hand I came across the raster package function , which calculates Moran's I just over the variable values (so either autocorrelation within the mosquito raster or in the land-use raster). Also sp.correlogram from the spdep package takes a variable vector as input instead of a linear model. The latter two functions give almost double the amount of autocorrelation compared to the first method.
So, I think I understand why you would check for autocorrelation in the residuals, but why in the variable input? Part of the correlation you're finding with the latter method might already be explained by your other variables as you're doing for example in linear regression? Are there options for doing something like a correlogram but with model residuals instead of variable input?
During summer days mosquitoes are hugely generated to become more active in attacking people. When we do hiking a lot of mosquitoes are there. We put anti-mosquitoes spray, but it does not work well. We are wondering how much it is possible the mosquitoes have ability to transfer coronavirse among people.
Does the mosquito larvae hibernate?
I am surprised that one week ago, I dried a pool full of larvae and sprayed some 95% ethanol. They have been dried and stayed immobile since then. But today, I poured some water, to my surprise, they are alive again! stretching legs and bending over!
I have a botanical extract which was prepared through maceration method. The powdered plant material has been soaked in pet-ether solution. The final product after filtration and drying is a very thick paste. I want to test this product against mosquito larvae. I hope someone who have experience working with pet-ether could share your knowledge. Thanks...
I have collected some mosquito larvae from different breeding sites in Egypt. Most of them are native to Egypt. I found in on place, larvae look like Culex torrentium according to the classification key of Harbach (1985) (saddle seta 1-x and seta 1 on abdominal segments 3-5). However, we know that this species is not present in Egypt. So, I appreciate if a mosquito taxonomist can look at the attached captures to tell me the right classification of this larva and whether it really belongs to Cx. torrentium or to Culex pipiens.
I will be rearing a colony of wild-caught Aedes albopictus at a remote field site where access to, and storage of fresh blood for artificial feeders will be inconvenient. It has been suggested to me that I should live feed the colony using domestic chicks (which we could safely/humanely house at the field sites), however I've failed to find a single protocol that describes this method. Has anyone used this method before, or know of a published protocol?
I want to deliver dsRNA into adult mosquitoes thorax using a WPI microinjection system. There were a lot of pre-pulled needles are available with varient inner and outer diameter. Can anyone suggest me, what will be the best needle size for the purpose of injecting adult mosquitoes?
Is it possible that Mosquito start acting as VECTOR for all or some strains of Covid-19,if not,why? any scientific/Biological reasoning in favour or against this thought.
The research idea is to study the fecundity and sporogonic development of wild mosquitoes. Apart from the low fecundity as a result of parasitaemia in the vector, could there be another association to look out for?
I am working on mosquito classification and identification and to do this I use many identification keys, I need to improve my skills and accelrate my work so Ican finish my work quickly and accurately, to do that I need to reduce the altering between the Keys.
A few months ago there was a chikungunya outbreak in some parts of Ethiopia. Chikungunya is one of the mosquito transmissible viral infections.
I want to use imageJ to count the number of eggs in Gypsy Moth egg masses, but I am not sure what camera and camera lens I would need in order to do this. I have seen a couple of mosquito papers that do this using a DSLR with a macro lens, but they do not provide any other specifics. Any thoughts or suggestions would be much appreciated. Gypsy moth eggs are ~1mm in diameter.
I am using Universal primer set (LCO1490 and HCO2198) to identify Culex mosquito species (tritaeniorhynchus and quinquefasciatus) by using COX gene. the expected product size was 710 bp but after gradient PCR. when i ran samples on gel i got two bands for each sample. i.e at around 300 bp and the other one was at around 500 bp. GC content for both primers is < 35%.
Good evening my name is Marta Albani, I am working at the Parasitology Laboratory of Sapienza University of Rome; I'm just working on insects increase number for a future male release after sterilization; I need methodological information about the blood feeding for captive females.
Dr. Marta Albani
I'm trying to figure out a way to ship mercurochrome-stained mosquito midguts for Plasmodium oocyst isolation. I need the samples not frozen. Does anybody have shipped them before? How did/would you do it?
The shipment will be overnight. We have tried with RPMI but the midguts were all mushy and unworkable at the arrival. We also tried with PBS and they looked a little better. But any better solution you can suggest is more than welcome!
We recently obtained some insect samples (Asphondylia) collected a few months ago by collaborators. The individuals were freeze dried, placed into pure ethanol, and shipped to us on dry ice.
I am having trouble getting a good DNA concentration from them (~50ul, 0.5ng/ul). I have tried the Qiagen DNeasy blood and tissue kit, phenol chloroform, and the Lucigen MasterPure Complete DNA & RNA Purification kit. I have tried varying incubation times (up to 17 hours) without any increase in DNA yield. I've been using the Quiagen Tissuelyser with tungsten beads to ensure thorough homogenization.
Does anybody have any protocols or suggestions on how we can get a higher yield? They are quite small insects (like mosquitoes), but I need a minimum concentration of about 5ng/ul in 150ul!
I am doing mosquito larvicidal activity against different concentrations of temephos to determine the resistance status in the field population. During the larval bioassay, I have been noticed that few mosquito larvae were formed to pupae and those pupae were live even at higher concentrations of temephos.
Please any one share is temephos has pupicidal activity or not. If not, why ?
Does anyone know what the contaminant in this nanodrop image could be? Its from a phenol/chloroform/isoamyl extraction of a single mosquito larva. I washed twice with an excess of chloroform. The ethanol precipitation was carried out using 100 ul of aqueous layer in 0.3 M sodium acetate and 1 ul polyacryl carrier in over 1 ml of 100% ethanol. The pellet was washed once in 70% ethanol and was a light grey color. I resuspended in 20 ul of water. Thanks.
- Rearing the target mosquito species for a susceptibility test is tedious task and expensive compared to intermittently catching the target species.
Some papers (eg Saudi Epidemiology Bulletin 1995; WHO report 1995) report the presence of Aedes albopictus in Saudi Arabia, but there could be a mixing up with Aedes unilineatus. Can anybody confirm the presence of Aedes albopictus in Saudi Arabia and in Yemen? Alternatively, can anybody send some specimens that look like Aedes albopictus for morphological or molecular identification?
I want to preserve my field caught dengue mosquitoes in RNAlater solution until RNA extraction later. Before that, I want to identify and sex them. Since identification of live mosquitoes would damage their body parts, i am going to cold anesthetize them at 4 celcius for about 10 minutes and do the sorting. Once it is completed I am going to preserve them in RNAlater and put them in the freezer. I want to know if it is ok to add the cold mosquito to RNAlater. Would it do any degradation to the RNA of the mosquito?
if you have any better option, please suggest.
We use Qiagen's QIAcube HT for RNA extraction from mosquitoes for our state surveillance of West Nile virus. We have spoken with other labs that do this, but they place their mosquitoes in microcentrifuge tubes and pipette off supernatant from those tubes. We, however, were convinced to use Qiagen's CMT racks (collection microtubes-racked) in place of microcentrifuge tubes as they can streamline the procedure by allowing the use of multi-channel pipettes and the ability to bead beat a full 96 samples at one time (the QIAcube HT can also pipette the supernatant directly from CMT rack tubes but that would not work for us for various reasons). The CMT rack tubes are much narrower than typical microcentrifuge tubes and we have to centrifuge the entire CMT rack (rather than individual microcentrifuge tubes) which makes it harder to produce high g-forces. These factors result in a supernatant that is not as clean as we would like. When those not so clean supernatants are ran through the QIAcube HT filters (we specifically use the RNeasy kit), some of those filters inevitably clog. We asked Qiagen if any other labs use the QIAcube HT plus CMT racks for mosquitoes but we typically received ambiguous answers or were connected to a lab that uses the QIAcube HT for mosquitoes but not CMT racks. Basically, I think we have realized that we are the guinea pigs for this particular combination. So I was curious if anyone else was having clogging issues with the QIAcube HT regardless of their starting specimen or protocol, and what they did about it? I will say about halfway through the WNV season I came up with a method that quickly gets rid of the clogs once they appear but I would rather they not appear in the first place. I found it was possible to wash the top of the clogged filter with the standing liquid atop the filter by carefully pipetting up and down with a 1000uL pipette (smaller tips are too pointy and could damage the filter). The particles causing the clog are not large enough to see with the naked eye but this process definitely redistributes them as the standing liquid almost immediately goes through the filter (the vacuum is running during this process). Thank you for reading.
I have conflicting information about the methods used to raise Anopheles funestus mosquitoes in the lab. Has anyone successfully managed this? Would you please share your experiences and/or protocols?
We have infected mosquitoes silenced for a specific protein with Zika virus . After a number of days we dissect the midgut, make a homogenate and try to check for virus replication by plaque assay.
Our plaque assay in the first dilution (1/10) shows complete lysis of the cells (and it doesn't look like plaques) in the second dilution (1/100) there is nothing. We have used just the midgut of non-infected mosquitoes to check if there was something in the midgut that would just kill the cells and nothing happened. We still have to make twofold dilutions... Thanks for thoughts on that!
A 1969 paper reports prevalence of Zika virus in Malaysia (Marchette NJ, Garcia R, Rudnick A (1969) Isolation of Zika virus from Aedes aegypti mosquitoes in Malaysia. Am J Trop Med Hyg 18: 411–415).
I tried to optimize conditions but havnt find any way. When i use mosquito cell line C6/36, target protein band comes at 50. But with live mosquito never got this. I attached membrane picture also please guide me how i can solve this issue Thanks.