Questions related to Insect
Recently, i have started working with SF9 cell line from thermo (Cat No. B82501) for which i was able to culture and grow the newly received vial in complete Grace's insect media at 27 degree Celsius in a non-humidified incubator as per the instructions (bright field images attached for reference). But i am not able to witness the viable cells after revival of frozen cells from previous batch. Approximately 10*7 cells/ml cells were frozen with 80% complete Grace's insect media, 10% heat inactivated FBS, and 10% DMSO as per instructions. Even though the doubling time is 24-30 hrs, there is slow growth of cells even after 7days.
Seeking expert advice regarding the possible reason for such slow growth of cells and how to enhance the culture conditions.
Cameron (2014) reports that the mitogenomes of 28 insect orders have been sequenced so far. Is there a recent publication that presents similar data for different taxonomic levels?
My research topic is to explore the biogeograpgic patterns of species richness of insects. I have the regional richness data of all insects and different orders from many locations. It's well known that insects include c. 30 orders with different numbers of species and phylogenies. I want to group different insect orders into several groups, and make a clear description of their diversity patterns. The problem is in grouping different insect orders into several groups.
I'm also looking for someone interested in this project. Please contact me if you want to join me.
I am involved in a project on biological control of the Comstock mealybug Pseudococcus comstocki in Switzerland. As part of this project, we are doing host specificity tests of a parasitoid and, besides P. comstocki, have tested so far the following non-target species: Pseudococcus longispinus, Planococcus citri and Phenacoccus aceris. We would like to test more species of the family Pseudococcidae and are looking for someone in Europe who could give us an identified starting colony for this purpose.
Thank you for your help!
I am working on my Tesis, and I need papers related with morphology and morphometry of cockroaches or any related insect (opthopteroids).
We purchased a new CG-MS and the provider suggested to use intrastent grade helium (to be cleaned with filters) instead of analytical grade He. I have always used analytical grade before and I am unsure of the impact of the swap on the quality of my samples (I mostly analyse plant and insect volatile compounds, some of which are present in minute amounts). I would be thankful for any technical advice you can provide.
With kind regards,
I recently deployed some insect pitfall traps and in several of them I unfortunately caught mice. How can I avoid this? I have seen suggestions of using shallow traps or small ladders, what is the best way to obtain quality insect samples and minimize risk to small mammals?
I have recently deployed some insect pit traps in the field, but I am concerned they may flood and overflow during rainstorms. Does anyone have recommendations for solving this problem?
Hi, I am looking for help emulating the process depicted in the image below. Essentially I am looking for a method to extract the juices out of feeder insects such as crickets, roaches, mealworms, etc in large quantities that ideally doesn't require any costly tools, without altering the juice in any physical (heating/desiccating) or chemical way. In the image below it was done without the use of a centrifuge, so I would like to avoid needing to purchase one. I'm aware that there are procedures for extracting hemolymph alone, but I need all liquid contents of the insects, not just the hemolymph. I tried using a juicer machine but all that did was sadly turn the insects into a paste. Any advice is more than appreciated, thanks!
I want to do research on some insects organs association of functional morphology, there I want steps and procedure to make research. Please help me. Here I have attached pictures for reference
Would the dominance of one of the microbes in the gut change if we give different foods to insects or larvae? Suppose the insect is supplemented with amylase-producing bacteria, fed with carbohydrates such as rice, corn, and other sources.
We are running qPCR experiments with cDNA samples derived from insect tissues using a QuantStudio Real-Time-PCR-System from Applied biosystems. Since a few weeks, we are facing severe problems with S-shaped or sigmoid amplification curves, leading to extremely decreased CT values for the relevant PCR samples. Intriguingly, comparing two technical replicates running in two adjacent wells, one sample is associated with this problem while the other one is normal. This is demonstrated in the attached figure that shows the amplification curves of two technical replicates: the one on the right hand is normal whereas the other is sigmoid.
The problem is not linked to a specific primer pair since it occurs with different primer pairs that have been used in the past without any problems.
Does anyone have a good idea how to avoid this problem in the future?
The targeted insects include fall armyworm and locusta migratoria. Need proteins study for further information. Suggest me proteins, database, or any research paper. Just share the specific insects proteins which are not involved in humans.
Hello insect cell experts,
I am currently culturing Sf9 insect cells for a protein expression and noticed some tadpole-like cells (red circled in the picture). They seem to reduce/disappear when cells are close to confluence. Are they just unhappy Sf9 cells, or could it be a contamination?
These cells are recovered from a cryo-stock and not yet transfected.
For the culturing, I use supplemented Grace's medium with 10% FBS and Pen/Strep.
I would appreciate if anyone with insect cell experience could tell what they are.
Thanks in advance!
I need some transparent medium for mounting small insects. I need a medium that a bit dissolves soft tissues and not needs complicated chemical procedure with mounting.
I am looking for a lab that can process insect DNA samples for me. More precisely, I would like to do hyRAD on my samples (they are not good enough for ddRAD). I am struggling to find one.
I am currently conducting an experimental study. I don't have any methods on how to count the generations that might be exhibited by the insect that I mass reared.
In our case it is impossible to traced from where is originated (species is clearly different from all others and belongs to group with limited distribution in Middle East).
I'm trying to get some good microscopic photographs of insect genitalia for my research work but I'm having trouble removing the air bubbles that comes inside the genitalia, no matter how cautious I'm.
Suggestions are appreciated.
How to make a model any one have experience? About insect sustainable waste management, economics value and market demand
I found this insect in the digestive tract content of Hypoatherina temminckii (marine fish). The sampling location in seagrass waters of Karang Congkak Island, Kepulauan Seribu (Seribu Island), Indonesia. I can't identify the insect groups, I just suppose this is part of Diptera but have never seen marine (or semi-aquatic) insects in my sampling location.
I'm trying to know where the holotype of Psammotettix confinis (described by Dahlbom in 1850) is conserved.
I searched informations on GBIF, INPN, EOL and internet, unfortunately, I didn't find anything.
Can you help me to know how can I find where the holotype is conserved?
I'd like to call your attention to browse and read a recent collection of papers on insect extinction, just published in Ecological Entomology: https://onlinelibrary.wiley.com/doi/toc/10.1111/(ISSN)1365-2311.insect-extinctions
I want to assess the similarity of insect communities within and between two groups of plants: a monophyletic clade and a paraphyletic grade.
Would you recommend calculating Bray-Curtis dissimilarities + Kruskal Wallis H test or a multivariate test such as MRPP or PERMANOVA?
Thank you for your help
How long will it take for arthropod ingredients to appear on our menu?
Recently, Nestle has released food for dogs and cats, in which, in addition to the usual chicken, they added chopped fly larvae. And no, the global corporation does not save on cats. Livestock is one of the drivers of climate change, and replacing cows with insects can reduce its turnover. Some insect products have been on the market for a long time. Tell us who you can try and what sensations to expect?
I need to preserve termite samples for future quantification of juvenile hormone in their bodies. Or extract the Juvenile Hormone and keep that samples stored for future quantification.
I may need to keep the samples stored for up to two months and they also need to "survive" an international trip.
Further disturbing data were published on the dramatic decline in the number of bees.
Now, in the media there was information that about 40 percent. Bees in the US did not survive the winter of 2018-2019.
Similar data is also found in many other countries.
This is very disturbing.
Is mankind able to solve this problem in time?
Will technological development solve this problem?
Apparently, a significant part of the bee population is killed not only in winter but also in other, warmer seasons. Also in the spring and summer, when large-scale spraying of crops with pesticides is used in agriculture, also used during insect feeding periods on flowers. Then many insects are poisoned and die.
How to solve the problem of a drastic drop in the population of bees and other pollinating insects?
I invite you to the discussion
Thank you very much
Case studies like real ones where the testimony from forensic entomologists was presented in court cases. I have searched but found the research case studies on the insect fauna associated with human cadavers like in the case studies below article for instance:
Anika Sharma, Madhu Bala, Neha Singh. Five Case Studies Associated with Forensically Important Entomofauna Recovered from Human Corpses from Punjab, India. J Forensic Sci & Criminal Invest. 2018; 7(5): 555721. DOI: 10.19080/JFSCI.2018.07.555721
I have been trying to dissolve TC-100 media powder obtained from US Biological in autoclaved distilled water to prepare 2X media for plaque assay.The solution is only clear at pH 4.5, as soon I make the pH to the desired pH (6.5) it starts to precipitate. I am using 1N KOH for adjusting the pH. Has anyone encountered a similar problem?? Can anybody suggest how the precipitation can be avoided?
Thanks in advance.
I am doing Iodine Test of insect's egg in flour for the first time and I just want to validate my results if what I have seen in the microscope is really the weevil's eggs.
The availability of relatively large number of disease resistant varieties compared to insect resistant cultivars suggest that incorporating disease resistance is relatively easy. This is also evident from the list of registered donors and available literature. Should it be understood that breeding for disease resistance is easier than that for insect resistance?
I have constructed a small hut with the support of bamboo sticks and were attacked by insects( mostly in black color)which made small holes all over the bamboos resulted in powder as in the attached pictures. Daily I see a lot of bamboo poweder and form dust throughout the room. Any remedy for this. Please refer attached pics. Thanks in advance.
We are looking for a suction tool/machine that would allow us to automatically count small insects (aphids, fruit flies) as they are sucked into a tube or container. It could be a counting with a laser cell, for example, or any other method.
To be clear: we would like to count aphids on infested plants, and one easy solution would be to use a suction/vacuum device (active sampling) so the insects would be counted as they are sucked into the device.
I have a problem with plasma membrane isolation from Sf9 cells, because after cells disruption and pelleting unbroken cells, cell debris and mitochondria the clear supernatant containing plasma membranes slowly becomes turbid. Something of white color is precipitating and makes difficult further steps. Does anybody know what is the origin of such precipitate and how to prevent its development?
I am developing a method to detect insect DNA in food samples via DNA metabarcoding.
I already designed primers (1 forward and 1 revers) for an amplicon of ca. 200 bp length that bind to mitochondrial insect DNA. Right now I am testing those primers in PCR to find the right temperature and conditions. I consider amplification curve and melting temperature from PCR and also bands on an agarose gel of all DNA-samples. All insect samples work well, but i have a quite unusual problem with honey bees:
They have a band at 200 bp on agarose gel, and there also is a melting peak at about 78°C. This is as expected and also like all other insect samples. But there is a difference: There are no amplification curves of honey bees in PCR.
I already tried cleaning the DNA extract with magnetic beads, that didn't help.
Additional information: I use EvaGreen as a flourescence dye in PCR.
Does anyone have an idea what could be the issue or what i could try to solve it?
I am happy to give more information about the conditions, if needed.
Thank you in advance.
I had been updating the old collection list from our museum and I found that there is some clash between family taxon for those three genera, some sources put them under Family Lonchodidae while some under Family Diapheromeridae.
Does anybody have an idea on how to successfully mass-rear Nesi bug? In full details, please! Any suggestions, thank you in advance.
the objectives of insect sampling in forest is to determine the abundance, diversity and habitat association of insect in forest ecosystem
I am having a very interesting issue with my protein expression. Currently, I am expressing a ~55kD protein recombinantly in insect cells (Sf9 to generate high titer virus and High-5 for protein expression). I used P3 of the recombinant virus to infect High-5 cells and have already verified the expression level of the 55kD protein via ELISA, WB and IFA. It is highly expressed in the lysate (insoluble fraction) of the pellet and some low expression was observed in the media. In addition the protein was observed to be in the High-5 membrane with IFA. Now since we do not have a SEC column in the lab yet, I was using the cell media to purify with the HisTrap 5mL column by Cytiva. After purification I got a single band at 55kD (it was not thick but it was visible) in 5 x 1.5mL elution's. Since the protein is in 500mM Imidazole solution I decided to combine, concentrate and Buffer exchange with HEPES Buffered Saline Solution for interaction assays using the Pierce Concentrators on my first try and then the Amicon Concentrators on my second try. I read on other posts here that the concentrator membrane composition might have caused the protein to bind, so I wanted to check that. I followed standard protocol supplied by the manufacturers and checked protein concentration with BCA at 595 and protein plate reader after. Both gave me a concentration of 2.9mg/mL and hence I loaded 1ug of the protein onto an 12% SDS-PAGE gel and even did WB with the antigen specific pAb. I got no bands in either assays for both concentrators (not even smaller bands)! I additionally checked the flow though material and washed the membrane of the concentrators but nothing. I am really lost as to how a protein can just disappear like that especially when the plate reader and BCA both detected protein? I was thinking maybe proteases but after purification and kept on ice all the time. My next steps would be to just purify from the insoluble lysate and use the SEC for desalting and separation of contaminating proteins, but I am worried that upon concentration the same thing will happen. I would be grateful for any advice that you might have that could possibly point me into the right direction. Thanks!
what method can be done for the extraction of plant/insect sample?
what method can done for separation and quantification?
for cyanogenic glucosides
I have a dataset of insect species and their abundances across sites (sites as rows, species as columns). I would like to determine compositional turnover between site-level assemblages (ie overlap) using the Horn similarity index. To do so, I would like to first calculate the beta diversity across sites using Shannon based on Hill numbers. Does anyone know of a package (along with a code) I can use to do this in RStudio? I have tried betadiver in vegan, but these do not give Shannon diversity indices.
I am wondering if there is a type of microscopy technique that people use in the field of insect physiology that allows muscle to be seen in living or prepared specimens.
Is it possible to run out BINs analyses on BOLD based on 16S sequences (wild bees)?
My fear is that BINs analysis is only possible for large dataset (such as CO1 sequences) but would not be relevant for 16S as insect species are not well represented on BOLD for this marker.
Any advice for diversity analysis of 16S for approx 100 species (up to 3 replicates each)?
I was thinking about looking at K2P distances, compare 16S and CO1 trees to validate our new barcodes.
Hi everybody , I hope all is well.
I've got a question
Could you suggest me some suitable pesticides for insect larvae in spirulina farm?
does any of you have some experience what is the best protease inhibitor mixtue for use in media for insect cells (S2,Sf9) please? I worked with commercial EDTA-free tablets based on inhibition of serine/ cysteine proteases and it didn´t work very properly. After FPLC I can still detect on SDS-PAGE followed by Mass spec. cleavage of my protein (small fragments of my protein). If you can recommend me some commercial mix of proteases for the insect cells media I would really appreciate it.
Good luck in your experiments and thank you!
I am interested in using RIP-seq or a similar protocol to identify microRNA targets in a non-model insect (Sarcophaga bullata). There is a published genome, but the annotated sequences do not include the 3' UTRs. My questions are:
How long is the fragment of mRNA that is generally pulled out using RIP-seq? Will I be able to identify the sequences I pulled out of this species using this genome?
Thanks in advance for your help!
I am wanting to compare fossil insect densities with charcoal concentrations and other indexes for disturbance.
My issue is that insect samples were subsampled according to geochemical patterns in the soil and are of varying resilution (between 2 and 8 cm) and also temporal resolution (20 - 120 years). The core where the other indexes from disturbance were counted from is from the same site but was subsampled in 1-cm resolution. We have a hunch that bark beetles and charcoal densities are related, but how to test this statistically, is it even possible? To correlate these insect time slices of varying resolution, with the data points from the other core?
Example insect samples:
S1 0-4 cm 2017 - 2004
S2 4-6 cm 2004 - 1969
S3 6-8 cm 1969 - 1946
S4 8-11 cm 1946 - 1899
S5 11-15 cm 1899 -1841
S6 15-17 cm 1841 - 1806
The results of my study are leaving me a little baffled. I would appreciate any insight you could give me.
What I expect: Larger objects (living or non-living) should heat up slower than smaller objects
My results: Heating rate is increasing with size, larger beetles heat up faster than smaller beetles
I tested for: The amount of water absorbed during the rehydration process does not play a significant role.
I am wondering: Could it be related to differences in fat/protein contents?
It is very common for Taxonomists to request insect specimens, from a number of institutions and museums, to assist them in their research on specific insect groups. It can take a few years to actually make use of these specimens and reach a conclusion with published descriptions. The institutions that make these loans in good faith, find themselves having to chase researchers for the return of the loans. I have heard of loans still not returned after 30 years and this may mean the loss of these specimens due to retirement or even death of the researcher. This issue is a BIG problem and so what would be a reasonable time for these loans to be made for? What ways can these institutions encourage researchers to return loans without threatening sending in the debt collectors?
I am working on insect associated mites. The parasitism and predation of the mites on the insect is evident from the direct observation and rearing experiment. I encountered a problem in confirming the predatory/parasitic relationship of the mite on the insect in molecular level. Could you please suggest a molecular technique for analysing the predatory /parasitic relation of the mite on the insect host. If we are going for a gut content analysis of the mite, which kind of molecular tests will be more sensitive in analysing it?
Can we use grace insect media
un-supplemented with 10 percent FBS only?
Is it necessary to add other suppliments?
I have tried to grow Sf9 with grace insect media supplemented with 10 percent FBS , antibiotics and antifungal but failed.
Please suggest me
I have small insect samples stored in ethanol and I need to obtain dry mass. I have access to a drying oven but I have also heard you can air dry them on the bench top in a lab environment with relatively consistent humidity. I would appreciate any tips.
I´m starting a pollination experiment. A cacti population will be selected for the study and maybe there are references to consider over an appropiate population, including some distance of human disturbance to pollinators.
I am planning to conduct a survey of patients who had taken an insect-derived drug given by a traditional healer in past years. I wish to know that if the medication was effective for the patients, any short or long-term side effects they know, and so that I can obtain some idea to study further on the medicinal value of that particular insect. I would be grateful if anyone can suggest to me what kind of questionnaire should I prepare, how should I analyze the data, and if anything else I should know?
I am interested in the size of Lepidoptera (moths and butterflies) and would like to know what is the smallest known species of this group of insects. It is probably a Nepticulidae (pigmy moths). The species in my figure below (unidentified) measures about 4 mm with the wings spread, and its dry body weight was 0.3 micrograms (0.03 mg).
We are generally more impressed by the higher figures (the oldest tree, the heaviest vertebrate…) than by the minima. Thus for instance one can read about the largest moths (Thysannia, Attacus: http://entnemdept.ifas.ufl.edu/walker/ufbir/index.shtml). However 'smallness' has interesting biological implications (see the recent book by A. Polilov 'At the Size Limit - Effects of Miniaturization in Insects'). I have seen descriptions of other nepticulids in the same range of size as 'my' species (around 4 mm: Dooren weerd et al.: http://onlinelibrary.wiley.com/doi/10.1111/syen.12212/full). Perhaps there are slightly smaller European species (some Stigmella spp., e.g.: http://lepiforum.de/lepiwiki.pl?Stigmella_Magdalenae).
So, does anybody know of any moth smaller than 3.5 / 4.0 mm?
Please, l need a clear experimental Proceedure for the extraction and purification of pigments from an Insect ( Weevils and Aphids)
Please can l get the step by step extraction procedure before the use of HPLC
Solvent Extractions, l guess maybe okay , but l am not sure yet of the adequate chemicals/reagents that maybe needed.
Finally as it is a pigment( Protein) do l need to pass any of the supernatant through indirect heating by use of water bath?
I look forward to your advise, suggestions, materials, journals and practical manuals that could be of help
I am getting to start a project on substitution of SBM with insect meal. The main aim of this study is to assess the effect of that replacement on performances of layers flock. I would like to know if anyone knows anything about mineral we should be careful with when using insect meal with black soldier larvae.
Catherine Roy, agr. M.Sc.
I would like to observe/ record insects behavior under webcam setting at night. So we are looking for some suitable infrared(Red light) lamps or IR LED to do this. Can anybody recommend us some proper products?