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Questions related to Gut
In my books on the subject of “mathematics of opinions” I have shown that even positive beginnings (must) end in complete decadence, unless there is a premature end. This is most pronounced in the area of politics, which for various reasons must degenerate into a melting pot of ideologically intellectually limited underachievers.
What is fatal, as can be observed live today, is the complete disconnection from reality: the leaders not only have an ideologically limited horizon of facts, they are also surrounded by a dense cordon of followers who absorb the leaders’ secretions with sexually fervent desire (I have also shown the background to this) and do not allow anything to get to them, i.e. they also effectively shield them from attempts at rational argumentation, which of course has an even greater effect on the ideologization.
On the other hand, the brain is bound to a demand-reward loop: people face challenges that, when mastered, cause a hormonal feeling of happiness (again, more details in “Mathematics of Opinions”). The absence of happiness hormones, in turn, causes people to look for new challenges. A problem arises when there are essentially no more challenges, because any action becomes without consequences.
This is also explained live in today's politics: normally, politicians should have to answer for an action, even if the responsibility only consists in asking themselves “Will I be voted out and can no longer act like this?” Today, the political system has reached a point where this no longer applies. All politicians, starting with the last backbencher in parliament, can do whatever they want without any consequences. A health minister can break laws and even the constitution, prove himself corrupt, send hundreds of thousands of citizens to illness and death and finally declare that all of this was not necessary - and stay in office and can carry on completely arbitrarily. No action has any consequences anymore, except perhaps for the last backbencher who is eliminated in the next round of elections.
Of course, this does not only apply to politics. The people behind it who still have some intelligence, the oligarchs or multi-billionaires who at least partially play their games behind the scenes, are also affected. For someone like Bill Gates, no action has any consequences anymore and the reward of happiness hormones is missing.
Ultimately, this leads to a displacement activity according to the motto "one man's joy is another man's sorrow". What can no longer be achieved through positive experiences of one's own can be achieved through negative experiences of others, for which one is ultimately responsible. The appeal of the matter is that you don't have to feel any consequences yourself, but others do (this is again explained in more detail in the books). This starts with the reporting of critics (even the followers in the justice system are subject to such mechanisms, as they too no longer have to fear any consequences if they ignore the law) and continues all the way to compulsory vaccination. I think it is quite possible that some politician or another has experienced a kind of inner reward orgasm in their uncompromising commitment to compulsory vaccination against the fears of those affected.
Where does it end? Wild conspiracy theories report cruel rituals by the rich and powerful in which women or - as the crowning glory - children are sacrificed. Well, if you think the development described through to the end, these are not conspiracy theories, but for some of those affected it may well end there. The ultimate kick from murdering someone else because something else no longer gives them a kick.
Is that also the background to the current desire for war that prevails everywhere? In Ukraine, the machinations of these people, who preach daily that more and more powerful weapons must be sent there, have probably already cost 1 million human lives, and another million could well be saved by what the Israeli government is doing, also with the help of the local hawks. And these things are not even far away: on social and other networks you can watch videos of people dying there at any time. Does that give you a kick? Or are we all already oversaturated with the virtual reality of the media industry, which can depict blood splattering around and guts hanging from houses in much more colorful ways with the help of AI? Is there really a desire to experience that live? Or is there still so much residual reality that you would rather not experience it yourself, but at least see someone you know personally die?
There is a lot to suggest that the ideological view is becoming increasingly narrow. It seems that the invulnerability of the warmongers leads them to make invulnerability itself an axiom of their thinking, in the sense that a war can destroy everything around them, but not them and their way of life. They seem to believe that they will continue to live in a luxury villa with servants who bring them everything they ask for, when everything around them looks like something out of a Mad Max film. And unfortunately, this virus also seems to be infecting those who are just the bulk of the claqueurs.
The reality: in the EU countries, no one has enough weapons to wage a war for even a few days. No one has a war industry that could provide replacements. The population is anything but ready or willing to fight; they are no longer even willing to work. One might consider the replacement of a private as warlord by a lance corporal as progress, but it is certainly not. The warmongering has no basis in reality, but is nevertheless constantly intensifying. Even if there is no war, it will still lead to hardship and misery. And although this cannot be overlooked, there is still a majority of the population that continues to enthusiastically participate.
As a consequence of current experiences, one view must be corrected: up to now, sociological and psychological models have assumed that a proportion of 25% - 33% of realists is enough to overturn the ideology. Current developments, however, show that an absolute majority, i.e. >50%, is actually necessary for a tipping point. Or, to put it more brutally: if you want peace, you must first kill everyone who wants war.
Hy, in my research I need to extract RNA from mice gut samples for further cDNA and qPCR. However, the results are really inconsistent, sometimes it's 700ng/µl, and for another replicate which has the same sample we get 350ng/µl, surprisingly it always seems that the first sample is the best and afterwards the quantity drops dramatically. We have found that it is really dificult to digest the samples since the gut samples are quite rubbery. The methods we are currently using is cutting the tissue in small pieces, adding Qiazol, trying to stamp the tissue with a micro mortar and vortex afterwards with a vortex for 30-45 min (in between vortexing we cool the samples on ice for 5 min). After phase separation with chloroform we are really carefull to only extract aqueous phase and premix it with 100% ethanol. Then we use the miRNeasy Kit for RNA isolation.
We can't think of any other reason for the inconsistency to be lying with the digestion method in the way we stamp and cut the tissue.
Also the other issue is that we always have low A260/230 ratios but really good A260/280 ratios. Perhaps this is due guanidine contamination.
Can anyone give some insight in what can be changed? Ofcoarse a tool such as a tissue ruptor with be handy, but we don't have acces to it at the moment, perhaps we can try liquid nitrogen, but this would be really time consuming when using a mortar and pestle (I need to do 85 samples).
These are mutually exclusive either for genetic, multifactorial or unknown reasons the immune system begins to attack a body it is designed to defend or
a problem of leaky gut syndrome where a permeable digestive tract allows access to pathogenic microbes inside defenses where they cause havoc colonizing arteries and brain tissue to provoking severe immune responses damaging nerves to bone? Your comments, ideas, and remarks are welcome. Where do you stand on this question?
Dear Colleagues,
I am working on histological observations of the gut of invertebrates.
My main research is on seasonal changes in the gut tissue of sea cucumbers.
This species ceases or decreases its feeding activity in summer and also decreases its feeding activity in winter.
In particular, the gut retracts or disappears in summer.
Gut retraction is recognised as a result of catabolising components in the body to conserve energy and to tolerate depleted stored nutrients.
Please advise if anyone else has researched this other reason for intestinal retraction in a professional manner.
Best regards.
Kai Tanaka
I am looking to amplify a region that can be used to group the gut bacteria into higher taxa like phyla or classes. The target has to be a region that is relatively conserved within the phylum or class but variable between these groups.
I hope some here who have done studies in this area can give me recommendations on which primer sequences worked best for you. Thanks!
The role of gut microbiota and diet has become a major concern for the health of Alzheimer’s disease (AD) individuals. The literatures shows that the microbiome composition has been linked to neurodegenerative disease and plays a critical role in the gut and brain axis. Microbial fermentation may release microbial metabolites that release short chain fatty acids such as butyric acid that has a major influence on the gut brain axis with effects on brain amyloid beta levels and plaque deposition. Researchers indicate that the understanding how the gut microbes interact with drugs and regulate gut microbes may improve the therapeutic effects and clinical outcomes of drugs with relevance to neurodegenerative diseases. The use of specific drugs may have major impact on the composition and metabolic function of gut microbiota metabolites and therapeutics for AD. Research is required on various drugs to assess drug-microbiota interactions on gram negative versus gram positive bacteria to reduce microbial metabolites such as toxic bacterial lipopolysaccharide levels in the plasma and brain with relevance to Alzheimer’s disease.
RELEVANT REFERENCES:
1. Sharma, A and IJ Martins. The role of Microbiota in the pathogenesis of Alzheimer’s disease. Acta Scientific Nutritional Health. 7.7 (2023): 108-118.
2. Vich Vila, A., Collij, V., Sanna, S. et al. Impact of commonly used drugs on the composition and metabolic function of the gut microbiota. Nat Commun 11, 362 (2020).
3. Wan Y, Zuo T. Interplays between drugs and the gut microbiome. Gastroenterol Rep (Oxf). 2022 Apr 8;10:goac009.
4. Weersma RK, Zhernakova A, Fu J. Interaction between drugs and the gut microbiome. Gut 2020;69:1510-1519.
5. Pant A, Maiti TK, Mahajan D, Das B. Human Gut Microbiota and Drug Metabolism. Microb Ecol. 2023 Jul;86(1):97-111.
6. Misera A, Łoniewski I, Palma J, Kulaszyńska M, Czarnecka W, Kaczmarczyk M, Liśkiewicz P, Samochowiec J and Skonieczna-Żydecka K (2023) Clinical significance of microbiota changes under the influence of psychotropic drugs. An updated narrative review. Front. Microbiol. 14:1125022.
7. Zhao, Q., Chen, Y., Huang, W. et al. Drug-microbiota interactions: an emerging priority for precision medicine. Sig Transduct Target Ther 8, 386 (2023).
8. Wang, S.; Ju, D.; Zeng, X. Mechanisms and Clinical Implications of Human Gut Microbiota-Drug Interactions in the Precision Medicine Era. Biomedicines 2024, 12, 194.
Hello,
my research group is rather new to the field of microbiome research. We have been collecting stool samples (stabilized in OMNIGENE gut tubes and then frozen at -80°C) and are now looking for a commercial partner for the microbiome analyses (16S rDNA amplicon sequencing).
We would very much appreciate recommendations for European companies to work with from scientists experienced in this field.
Thank you very much!
Dear all,
This is a microplastic image from the fish gut, and I am having difficulty identifying this shiny particle. I am guessing it may be a film particle. If someone knows, please guide me.
Thank you so much in advance.
With Regards
Pragya Mehta
"A high-fat diet promotes cancer progression by inducing gut microbiota–mediated leucine production and PMN-MDSC differentiation"
How do specific metabolic and immune factors within the gut interact with the reproductive tract, and what are the precise molecular mechanisms governing this interplay, considering the impact on reproductive health and potential implications for fertility and pregnancy outcomes?
I would like to cultivate lactobacilli in an intestinal organ-on-chip model and stain it with a suitable dye either beforehand or, if necessary, after the end of the experiment with a suitable antibody for immunofluorescence microscopy.
Briefly, I would like to check the Lactobacillus attachment/localization to/in the intestinal tissue.
Is there anyone with experience in this area and could explain possible procedures?
Thank you very much in advance!
I'm working in a project where I need to test the enzyme activities of an insect's gut fluid. I've already tested it and found some good activity. Now I want to know whether the enzyme, showing high activity, is endogenous to the insect, or just produced by gut microbiota. There is no genome sequence or any other information available about that insect. How can I test it? How can I prepare enzyme solution, containing only endogenous enzyme of the insect?
I've read about the antibiotic feeding process. Anything else?
I found a paper where they incubated the insects for 24 hours, dissected, collected the gut fluid and then filtrated the gut fluid through 0.22 micrometer syringe filter, then demanded that this filtered solution contain no bacteria, neither their enzymes. But how can they demand there is no bacterial enzyme? If it is for starvation, then isn't it obvious for the insect's enzyme to reduce too? Can anyone provide any reference on support of this method, or concept? Please let me know.
I'm struggling in this issue and can't find any solution. I can't use the antibiotic method for a reason. Please help me out. Thank you.
I am working with a marine non-model organism that has a planktrotrophic larval type and I found out that the larvae are not very selective while feeding. I am planning to do RNAi by feeding them bacteria (E.coli) expressing dsRNA. As a preliminary experiment, I am planning to feed them with bacteria for three hours. I would like to know whether or not they are ingested the bacteria. Is there a method to stain ingested bacteria? The larval type of my organism is transparent and the gut can be visualized using an optical microscope.
Introducing a Super Unified Theory of SU(11) of 10-dimensional Space-Time of the physical universe instead of four dimensional Einsten's Space-Time of GUT SU(5) of SM, which explained the forces of Strong by SU(3), Weak by SU(2), Electromagnetic by U(1)and a new fifth force called latent force by the new particles of SU(6). The strength of SU(6) are so large that it can changes the exotic matter fluid into ordinary matter then everything.
Hi everyone,
im using this established protocol:
I already optimized everything, written under "troubleshooting" in this protocol. I directly obtain the samples from the endoscopy, its less than 20 minutes between extraction and starting work in the lab. During this time the samples are on ice and in the media recommended in the protocol.
So far i was only able to generate organoids from childrens biopsies, but not from older patients. It seems, as if they needed a stronger Wnt-activation?
I use recombinant human Wnt3a at the correct concentrations. Would you recommend switching to Wnt-surrogate or Wnt-conditioned media?
The protocol should also work with samples from older patients.
Is maybe the tissue piece from the endoscopy too small and i need larger samples from the surgery?
Any other ideas, what i could try?
Thank you,
Marco
Dear Joanna,
I just saw your nice video contribution about your study about acupuncture as treatment in diabetic neuropathy - great: I wish you a great success! Ich hoffe, Dir geht es gut: Berlin macht sicherlich weiterhin viel Spaß inmitten Deiner Familie und an der Charité. Bei mir sieht es ähnlich gut aus, es gibt so viele Dinge hier in Dresden und Leipzig - wunnebar:-)
Wünsch Dir eine tolle Zeit in 2021, bis denne und liebe Grüße,
MIchael
I presently use the powersoil pro kit for the extraction but I am not getting my desired result maybe cos the starting material (Daphnia magna gut) is too small. i'd like to know what methods has been used that yields a good result.
I'm currently working on probiotic adherence to human's gut and, while writing the manuscript, received a question from one of assesors whether there is a minimum adherence inhibition? Maybe in 30% 50%?
As far as I read papers, none has ever mentioned.
I want to isolate Bacillus subtilis from animal gut digesta and was wondering what is the best medium to grow it effectively. is it TSA? also what are the conditions of the growth, aerobically or anaerobically?
Thanks
Dear Friends,
Please, join a discussion on Theory of Universality in Physics, a comprehensive GUT. The discussion is live in Academia now.
My Theory is proved to be experimentally correct on 4 aspects, namely :
1. The increase calculations for Sun (both radius and mass).
2. The increase calculations for Earth(radius and volume)
3. Dark matter speed, u
4. Gravitational constant, G.
Kasibhatla Surya Narayana
I need to purify DNA and RNA from frozen GUT tissue (rectal biopsy tissues from monkeys mainly). The tissues are frozen at -80°C with or without RNAlater. I am thinking to use TissueLyser, since it will be safer when using infected tissues. According to the protocols, if the tissue are stabilized with RNAlater, I can thaw it at room temperature and proceed with next steps for disruption and homogenization. What about frozen tissues without RNAlater? How to take and weight a tissue, if it is frozen in the medium at -80°C and should not be thawed (according to the manuals)? What size of stainless steel beads would you recommend? (I am thinking to use 5 mm beads). I am also not sure what TissueLyzer would be better to use...
I will appreciate any suggestions related to the use of TissueLyser for rectal tissue samples.
Thank you everyone in advance!!!
Which are the relatively more successful refutations so far of the foundations of quantum physics? How do they compare with Relativity when it comes to efforts to build up a GUT on bases that may apply also to many other sciences?
I have already processed my fish gut samples for microplastics analysis from extraction to filtration and drying. The extracted microplastic particles were stored in dried glass fiber membranes. Is it valid and possible to detect PAHs traces by using the membranes alone and directly analyzing it with gas chromatograph or HPLC? Will the retained PAHs can be detected if I carefully pick the particles and testing it directly with the gas chromatograph or HPLC? Thank you in advance.
Hi, I'm searching for studies on the diet of the mayfly Ephemera vulgata (Linnaeus 1758)? From a European database (freshwaterecology.info), its feeding type preferences is given as 20% gatherer (FPOM sediment) and 80% active filter feeder (FPOM, CPOM and micro prey from the water column). The reference for this information is given as the AQEM expert consortium. I would be really interested to see the original study providing this information, or any other studies that explores food item preferences (e.g. gut content) of E. vulgata. If you know about any papers, thesis, reports etc. with such information, even anectodical, I would be grateful if you could share this information with me.
Tor Erik
In bee pathogen diagnostics, we often use different sets of primers to identify pathogens in bee guts.
One of the primer sets we have been using a while in PCR, we are now using to quantify pathogen loads via qPCR.
However, we discovered that a frequently used primer set, published years ago and used frequently in different papers, is showing a strange curve. It amplifies a fragment of the 18S rRNA gene, of Microsporidian pathogens belonging to the genus Nosema.
We tried different conditions and concentrations, but the curves stay the same.
In addition is a BioRad data file with the standard curves, and a picture for those who cannot open these types of files.
Efficiency is always low when using these primers, while R² is high, so we think it is due to the weird curves.
Does anyone know what could be the reason for the amplification curves?
Or the reason of the low efficiency?
Thanks!
More info:
Primer info:
Forward = Nosema universal, edited by Fernandez 2012, originally from Higes 2006 (TATGCCGACGATGTGATATG)
Reverse = Nosema universal, from Higes 2006 (CACAGCATCCATTGAAAACG)
qPCR Program:
2 min at 95°C, 39 times: 15 sec at 95°C, 30 sec at 56°C, 45 sec at 72°C
Fernández, J. M. et al. (2012) ‘Asymptomatic presence of Nosema spp. in Spanish commercial apiaries’, Journal of invertebrate pathology. J Invertebr Pathol, 111(2), pp. 106–110. doi: 10.1016/J.JIP.2012.06.008.
Higes, M., Martín, R. and Meana, A. (2006) ‘Nosema ceranae, a new microsporidian parasite in honeybees in Europe’, Journal of Invertebrate Pathology. Academic Press, 92(2), pp. 93–95. doi: 10.1016/J.JIP.2006.02.005.
Dear colleagues! We plan to isolate mitochondria from freshwater amphipods, but didn't find any methods in literature - the closest found was the method of isolation from whiteleg shrimp Litopenaeus vannamei.
The problem is - the amphipods are quite small - around 1 cm long, so it's hard to isolate the gut before mitochondria isolation.
Will it work if we use just the sample of 10 g (or is that too much?) of amphipods and blender to homogenize it in isolation medium? Or it is crucial to select only some parts - for example only the amphipods legs and antennas?
P.S.: we do not have chitinase, nor the chance to get it in time.
Hi,
I need to compare the alpha diversity of three communities (the gut bacteria; the gastro-intestinal helminth community ; the bacteria community found in the spleen of rodents)
For the gut bacteria, I have abundance data
For the two other communities, I hace presence/absence data
Alpha diversity : I was thinking of using richess and Shannon index, for all communities (even the two with presence/absence data only). I know that Shannon index is made for abundance data, but is there any statistical problem in using it for 0/1 data ? It would be a shame to analyse gut bacteria using only presnece/absence data and richness index for all communities...
Beta diversity : I was thinking of using Bray-Curtis distances for all communities. I know that Bray-Curtis is made for abundance data, but is there any statistical problem in using it for 0/1 data ? It would be a shame to analyse gut bacteria using only presence/absence data and Jaccard distance for all communities...
Thanks,
Nathalie
probiotics can be used for controlling pathogenic microorganism in the gut of poultry, but what else these probiotics can be used in poultry
The research topic is for final year graduation from bachelor's degree. I had a topic in mind such as Leaky gut syndrome: The imbalance between microbiota and
harmful bacteria, but due to unavailable equipment in our labs, I need other topics. Your help is appreciated.
If I’m looking at the association between cardiovascular disease (CVD) risk and gut microbial beta diversity, I understand that this would show the association between levels CVD risk and inter-individual differences in gut microbiome composition. However, I’m finding understanding that association in a biological context a bit tricky. For example, if it is a positive significant association, is that suggesting that harbouring a more distinct gut microbial community compared to somebody else , may be linked with a CVD risk (i.e there is no “core” microbiome signature associated with CVD risk per se that would be common across individuals, more likely to be different compositions when comparing).
I'm more familiar with comparing beta diversity between disease vs control, rather than association with an endpoint, so any explanations would be appreciated - thank you.
I stained dissected gut of Drosophila larvae using DCF-DA for visualisation of ROS. Gut tissue was fixed with 4% PFA overnight at 4°C. Under fluorescence microscope, bright red fluorescence was noticed insteed of green fluorescence. Is it normal ?
Hi,
I am trying to study the gut microbiota of honeybees. Is it necessary to separate the host tissue (gut particles) after we homogenize the gut? Our lab is using DNA amplification of 16S rRNA and the culture method for the study. I am curious if the host tissue has an impact on the bacterial DNA extraction process and/or in qPCR analysis.
As we have known, the bacteria such as E. coli and P. gingivalis can secret LPS, EVs to induce the immunogenic response of the host. So, I wonder if there are other components that can be secret into the gut by bacteria? Thanks very much!
This would include tools for assembly, binning, and annotation. If it can be of any help, it's for gut samples.
I am running RNA extractions on whole gut samples for downstream RNAseq. For one individual I realized there was a length of gut tissue still in the original collection tube that I didn't add to the homogenization solution. I'm not sure what region of the gut it actually is or proportionally how large it is relative tissue that was homogenized (it is smaller), but I'm worried that if there are regional differences in RNA expression profiles that will bias the RNAseq data towards the already-processed portion of the gut.
Is this sample salvageable? If I extract RNA from the leftover tissue, could I just combine the total RNA sample volumes from both prior to sending in for sequencing? Alternatively, if we sequence both separately could we normalize and combine reads somehow? Are there any other strategies that would be more robust to prevent bias? Thanks in advance.
I have metagenomic data of gut bacteria of tribal population of Himachal Pradesh, can anyone help me regarding analysis of same for writing a research article. we will share authorship for the same.
Can anybody help me with the protein extraction from the gut of adults and Larvae of Zebrafish?
I have a problem: I can't get a good result in Western Blot with adult zebrafish gut or with zebrafish larvae (5 dpf). When I extract the proteins, the bands on SDS-page appear not very concentrated and after the transfer, when I paint the membrane with ponceau red, the result is not clear and there aren't defined bands , but everything is very blurred as if the loaded samples were dirty. The protocol we use provides for the homogenization of the sample with lysis buffer
(SDS 1%, EDTA pH 8.0 10 mM, Tris-HCl pH 8.1 50 mM, PMSF 1 mM, protease inibitor) or Ripa buffer (Sodium chloride 150 mM, Tris-HCl pH 8.0 50 mM, Nonidet P-40 1%, Sodium deoxycholate 0.5%, SDS 0.1%, protease inhibitor). Then I heat the samples for 5 minutes and centrifuge them at 12,000g for 15 minutes at 4°C. After, I load from 20 to 50 ug of protein in each well in a 10% or 15% gel. I do either dry or wet transfer, but either way it doesn't work well. What could I do to improve my homogenization of samples and transfer?
We have procured metronidazole powder ( active ingredient) for an aquaculture experiment. The purpose is to study the effect of metronidazole on gut flagellates. But we have a problem with dissolving metronidazole in pure water.
Could someone suggest how it could be done?
Regards
With regard to patients suffering form spontaneous bacterial peritonitis, it seems counterintuitive that those patients tend to NEVER have anerobes as the causative agents despite the common leakage theory that should allow anerobes (that fill the gut) to leak whenever the opportunity is present? Is the leakage theory a sloppy theory? Do we need to tinker it a little bit? Should we adapt a new theory?! Or, should we go playing more in our labs hoping to uncover the real etiology and decipher this anerobes enigma?
Hi, I'm trying to take picture after immunolocalization with insect guts. After I did all process, I have a problem when I mount the guts. I used pipette and pipette tip (I cut the tip) to do this. But, most of the guts stuck to the tips.. Do you have any idea to solve this problem?
I have assessed the effect of trypsin and pepsin on phages. Thus, I want to compare the normal concentration range with the concentration where phages remain viable.
Hello everybody
I am facing an issue in fish gut digestion using 30% H2O2. After the digestion process, I am getting a fat oil layer that might trap plastic particles. Kindly help me out with this to digest the oil layer or separate particles from it.
i need help regarding designing oral feeding bioassay for adult beetle (scotinae, coleoptera). my study involves profiling the effect of a toxin on midgut of the insect through transcriptomics. i have tried some semi artificial diet based protocol. but it did not work and i am unable to find much literature on it. please suggest any other way to feed the insect so as the chemical reaches the gut.
i have been working on adult beetles. can i also use lower developmental stages (larva or pupa) for such a study.
thank you :)
I want to study the sorption and desorpotion of an antibiotic by microplastics in the gut of a marine fish species. I'm wondering if I can prepare a solution that is extremely similar to the intestinal environment (simialr pH, compotiosion of ions, disgest enzymes and etc), and then do the kinetics test in that solution? Has anyone done a similar study of this? Could you kindly share the protocol to prepare the "intestinal environment-like solution"?
(I speculated that the intestinal contents in fish are more like liquid than a solid? Am I wrong?)
Thanks in advance!
I want to extract the midgut of Callosobruchus maculatus larvae to isolate their proteases. This will be my first time dissecting a small larva (2nd/3rd instar). I read some publications about how they dissect and prepare the gut homogenate. However, some research protocols were different from the others. Sometimes, the insects were dissected by only using cold distilled water, but others by using buffers (citrate-phosphate or sodium-acetate) or NaCl solution.
I am following Karam et el (2017) method on digestion at 40C instead of 60C as most publications reported disruption in the plastic components. KOH had the most optimum digestion as well compared to other bases and acids by having the least damaging effects on the components. Im following 1:3 ratio (organ:solution). Most of my samples are cloudy. How to increase the digestion of my samples? Thank you.
I want to isolate the microbiom from the gut of the larvae but I have problems with the dissection. I tried to use needles and a scalpel from a dissection set for biologists but I had problems to open the larvae and to find the gut. Is there any trick?
For example: The intestine/gut of zebrafish as model organism.
I would like to study metabolic profiling by using GC/MS. Could you please suggest suitable protocol for determining metabolites in tissues.
Thanks in advance.
I'm looking at gut bacteria in heat stressed birds but I'm not certain after how long can I sample faeces after episode of heat stress. I don't want to sample early before the changes shows in faeces or too late.
Dear Colleagues,
I have faced a challenge in the process of primer design for Real time PCR.
Two main bacterial populations exist in our gut, firmicutes and bacteroidetes. I am wondering how to design a specific primer for such wide populations? what genes should I target for primer design?
I appreciate your help and guidance because I really need that
Thanks
Hi, we found some fish parasites in the gut of Serranus cabrilla, and in the gut and on the skin of Thalassoma pavo? Does anybody have a clue to which groups this parasites might belong to?
Thank you very much in advance
Hi all,
My experiment is to find the differences of micro bacteria in gut of marron Cherax caini. there were 2 groups of marron , one fed every day, and one fasted for 4 weeks. At the end of trial, marron gut were applied RNA gene sequences.
Our lab performed some 16s rRNA sequencing works recently, we acquired more OTU/ASV for oral samples than feces samples. We wonder if this result reliable or not, but we don't find there are any papers indicated the gut has more microbiota than the oral cavity. I will be very appreciative if you can give some valuable suggestions.
Earthworms were stored at minus 80 degreed freezer. I need to extract the gut from earthworms for microbial analysis. However, i am having problem to extract gut because the earthworm became wet from inside after defrosting and its really hard to pull out the clear guts.
I have incubation several pots of earthworms and I want to find out my treatment effects on earthworm gut microbiota. But I need to go back to China next month and I can not extract the DNA of earthworm gut right after I finish my incubation due to the condition limited. So I need to freeze dry the gut sample and take them back to China for DNA extraction and analysis. Could that be possible?
To understand the gut microbiome-brain axis
I need to compare the human gut microbes with human to identify functional genome similarity. Is there any online tool for comparing the genome between different species?
I did the hemolytic test, The result showing after 24 hours indicated that the bacteria have gamma-hemolytic (non-hemolytic), but I kept the plate in the incubator for 48 hours, then the clear halo zone showed around the colony (Beta-hemolytic). So which result should I use? I read some papers and normally they did incubation for around 16 -24 hours.
Another found was that 75-90% of my isolates from the fish gut have Alpha or Beta hemolytic. and almost 100% of my isolates with antibacterial activity are hemolytic strains. So I would like to ask two more questions:
1. Is hemolytic really a necessary safety test for probiotic due to many bacteria with hemolytic is available in the environment and we still can scope with them?
2. hemolytic activity and antibacterial activity are in correlated or not?
Thank you very much for your suggestion and comments!
Good day. I am working on collecting larvae samples of a saturnid moth (Imbrasia belina), of which i would like to use for DNA barcoding and SNP genotyping. How do i prepare the specimens to avoid contamination from the gut contents? do i have to degut the larvae prior to preserving in absolute ethanol? so as to extract good grade DNA?
Many thanks in advance..
I have dissected 2-3cm intestinal segments, flattened and stretched on a platform, fixed in 4% PFA for 24 hours, and dehydrated in 0.03% sodium azide for a maximum of 4 days (is this too long?). After this, what are the best solution and temperature conditions to store this tissue if needed for immunohistochemistry?
Also - what is the best method by which the myenteric plexus and submucosal layers of the gut can be isolated from these whole mounts for histological staining? Thank you!
Since its emergence in 2019 the SARS-CoV-2 remains a challenge for scientists, clinicians and the overall population to solve. Besides its rapid spread and harsh respiratory distress it did induce, further secondary outocomes are observed. In particular, recent works highlifhted a preferential accumulation of its spike protein in skin tissues, specifically sweet glands. It appears that skin epithelial cells could also be a secondary host-cell of the viron.
this instigates several questions including:
i. did the viron targets all epithelial cells, such as skin and gut epithelia?
ii. if so, does it really rely on angiotensin converting ezyme receptor abundance in these cells? would these cells act as reservor for the virus ARN?
iii. could spike protein modify the skin physico-chemical properties?
etc.
I really wish to have a deep discussion of this fact
Hello everyone. I have collected intestinal biopsies from stem cell transplanted patients in RNAlater and wish to quantify neutrophils only via qPCR. As we know that the mucosa of gut is filled with several immune/epithelial cells, I want to target just one cell population. Is there any singular magic marker for neutrophils? Please let me know. Thank you in advance.
I am comparing the gut microbial diversity of 2 groups of individuals. I found that the alpha diversity is significantly different but the beta diversity is not significantly different. Can I still assume that the groups are different in their microbial population?
I want to evaluate if some groups of humans have endotoxemia or are at risk for. So, I have this huge doubt whether I must choose serum LPS ou serum LBP or even other marker/protein, because the research papers do not discuss what is the bettter marker for diagnosis endotoxemia, or the better marker (from gut) for activating serum monocytes.
Anyone who has a protocol to study microbiota? How to maintain the guts, etc?
I want to begin studying microbiota from aged rats and I really know nothing about how to do that.
Fecal microbiota transplantation (FMT) is the administration of a solution of fecal matter from a donor into the intestinal tract of a recipient in order to directly change the recipient’s gut microbial composition and confer a health benefit.
I heard about the beneficial using of bone broth that is rich in collagen for healing leaky tight junction in gut; but I did not find scientific evidence about it, may you help me to find scientific evidence and studies ?
Recently I have been interested in microbial communities from gut & intestine of the same fish species popular in Indonesia marine aquaculture, Grouper (Epinephelus sp.), but are raised in different farm.
For example E1: monococcus 36%, streptococcus 29%, ... bacteria 70% gram negative ... etc
compared to E2: monococcus 31%, streptococcus 35%, ... bacteria 63% gram negative ... etc
I am working on the gut bacterial diversity of thrips and I sequence 16S V3-V4 regions of one species of thrips with the larvae pupae and adult life stages.To my surprise, I have 99-100 % bacterial genus Arsenophonus in all my samples. What may be the possible reasons for this. Do this bacteria replace the other bacteria and what experiment does I need to do to confirm this?
Please share your preferred host DNA depletion method for microbial shotgun metagenomic sequencing. Please explain why. If your preferred method is to sequence everything and remove host reads through bioinformatics, share that too.
Some context: I will be sequencing degraded formalin-fixed ethanol-preserved DNA collected from amphibian gut swabs. Debating if I try one of the chemical removal methods or simply deal with this computationally. Most of the methods I have found are setup for removing mammal host DNA, so not sure of things like removing differentially methylated DNA, etc. I am open to trying new and developing methods for this.
Good morning,
for our project we will dissect bumble bee trying to obtain and separate the external with the internal part (gut),
do you have any suggestion or tip do it?
we tried just after took the insect from the -80 °C and dissect on ice, but the terminal part was tough to remove and consequently the gut was contaminated with the external portion, we will try other times, if you have any suggestion, we would appreciate a lot!
thanks
I read some articles on mollusc and fish diet analysis. The formula for percentage frequency of occurrence stated above:
%F0 = Nei/Ne x 100
and percentage numerical abundance :
%NI : Ni/N x 100
especially in mollusc. I want to know how is the calculation for Nei, Ni and N take place, is it an individual organisms count under microscope slide or do we need to used specific device or equipments such as haemocytometer or sedgewick rafter to count the number of the cell per area before we can fill in the Nei, Ni and N in the formula?
Thank you.
the gut related diseases like White feaces, white muscle, white gut are doing much damage to shrimp by retarding growth and increasing FCR and eventually heavy losses. is there any medicine or cure available now apart from preventive measures?