Milk - Science topic

Changes in the diet of humans as well as cattle feed. Plus, converting animal and dairy wastes into bioenergy.

This question has not a single answer.

Milk yield depends not only on nutritional regime but also on genetic merit, environmental conditions, body condition at the beginning of the lactation, etc. NRC and INRA give comprehensive methods to estimate requirements of dairy cows. The rationing would also depend on the ingredients (forages and concentrates) available for you.

Surely, HPLC is better and more accurately

Dear Helia

In the Gerber test H2SO4 is used to increase specific gravity of milk serum which makes greater difference between milk serum and fat globules. It also destroys stickiness of milk by dissolving chemical bonds (hydrogen bounds first).

Adding sulfuric acid will denature the casein. As a result, the casein molecules are likely to stick to each other and form a centrifugable pellet that goes to t,he bottom of a centrifuge tube after spinning.

Sulfuric acid is very reactive and dissolves most metals, it is a concentrated acid that oxidizes, dehydrates, or sulfonates most organic compounds, often causes charring.

You can try out this one (think of personal safety and protection): take a lump of sugar and put 2-3 drops of 91% sulfuric acid and watch. The sulfuric acid is extracting all the water and leaving a black, charred residu.

Milk is an oil-in water emulsion and thus, consists of both water and fatty acids (short-chained fatty acids, unsaturated, saturated, polyunsaturated etc.) The lipids are uniformly dispersed throughout the milk. Here, milk proteins (such as casein and whey) act as emulsifying agents that contributes to homogenize the lipid droplets in the milk.

One alternative to measure the milk fat content is to utilize the UV-spectrometry since milk fats are able to absorb UV-light, if I have understood your question correctly. You can read more about this by searching for milk fat content measurement using UV.

Hi Luis. I don’t know any good programs but beware of calculating a water footprint for milk, or any other animal product for that matter. For all of the examples I’ve seen the largest majority of water is used in the growing of the feed for the animals.

If you are in California or Mediterranean Europe then the source and cost of irrigation is a strong consideration. If you are in North East USA or Northern Europe, the water needed falls as rain so it is of less consequence.

First you should determine fat content in butter. The rest is butter plasma. Example: fat content is 80% (butter plasma is 20%) and total acidity is 0.05%. The acidity of butter plasma is 100*0.05/20 = 0.25%

You can use zeta potential and Particle size distribution or rheometer ( rheology methods). Also, you can use the oil off method.

What about the homogenization of the skim milk at the first step?

Kindly see the RG link below.

You might also look at the illustrated calculation in the Handbook of Food Engineering Practice

One of the elements influencing the sensory qualities of milk and meat products is the kind of forages consumed by ruminants. The impact of the type of forages (corn silage, grass-based forages), the conservation strategy (hay versus silage), and the botanical composition of the grass on the sensory qualities of cheeses (colour, texture, flavour) are discussed for dairy products. There is no scientific evidence that genetically modified feed has an effect on milk output or composition. It is crucial to remember that cows consuming GM food do not modify an animal's (or a person's) DNA. Cow's milk has no GMOs. A contrast may be made to diabetics who use insulin; the individual does not become GMO. A dairy contrast is that a cow given chocolate does not produce the milk that comes from that animal.

Sravya Jasthi Hope this answer is relevant to your query.

Baby Bottle Tooth Decay can occur when babies are put to bed with a bottle, when a bottle is used as a pacifier, or if a baby uses a bottle or sippy cup for extended periods of time.

Green color is perhaps due to the presence of both semi-oxidized (blue) and completely oxidized (yellow) form of TMB. We observed green color when oxidized TMB by nanozymes in a buffer with ph lower than 4.

Hi,

you mean you have two bands within one sample when using BSA? One at around 25kDa and one at 35/40kDa? Or in different samples?

In my experience BSA leads to more unspecific binding than blocking with milk.

In case it is within one sample then I would trust the BSA more with the band around 25 kDa and the one above is possibly a non-specific signal. Do a proper control without GFP transfection and maybe also try blotting with secondary antibody only without primary antibody detecting GFP, because secondary antibodies can lead to unspecific signals as well.

Also, kindly check the following link that may be useful:

Not safe if the dog is infected with zoonotic diseases like PPR

your protocol looks fine to me but I would advice you to check your buffers even though tailing effect of your bands might be probably for the dye you are using or the protease inhibitor

it would be helpful if you show some pictures.. how is your ladder separating in the gel? is it fine and well separated?

what is your housekeeping antibody, do you have a picture for that?

In my experience (and others I know as well), membranes probed with the NIR LiCor secondary antibodies do not strip well at all, especially at if any point in your process the membrane dried. LiCor sells a stripping buffer that is supposed to work for their antibodies, but I have never tried it myself.

The fat globules are known to block measurements in milk because they have a large volume in the liquid and sometimes cover other elements of the milk.

My immediate thought was whether MSG has been shown to be a substrate for BCRP (Breast Cancer Resistance Protein) that is known to excrete drugs into breast milk. I've not looked at the literature to bolster or refute this possibility.

As a postdoc back in the late 1980's and early 90's, the lab protocol was to dilute the commercial antibodies in 1X PBS at a 1:10 dilution and store them at -80 or -20 in 20 ul aliquots. The antibodies lasted mostly for years. Its a practice that I have kept using. I just got a new shipment and I am open to change.

60ug is a lot of protein. I have the same question about reducing agent. I would reduce the amount of protein you load as that is likely contributing to the smearing you see. Does CDH4 have a lot of post translational modifications? That could retard the migration on an SDS-PAGE.

Dear Marc,

Thank you for your kind reply. We would like to set up the HPLC method for analyzing milk cortisol but seems that HPLC-FLD is required (we don't have it). Also, we don't have LC-MS equipment at the moment. However, I appreciate it if we can communicate via email to further discuss this. Would you please provide your email address or send me an email to: jalilgh@konkuk.ac.kr either more convenient for you.

Thank you

Hello Rogelio Valadez-Blanco ,

I didn't explore exopolysaccharides' possible effect on whiteness in the literature. Thank you for your interpretive answer. I will search and think about this perspective.

Thanks and Best Regards,

Hi Samvit

We use non-fat milk to block our membranes but do so in TBST (TBS + 0.1% tewwn-20). The detergent helps to keep the milk suspension stable and can help to avoid it from precipitating out (forming lumps that stick to membrane).

We block for 1h at RT. Moving between 4C and RT often or not allowing buffers to calibrate to ambient temperatures can also accelerate precipitation.

Hope this helps

Aparajita

These publications also probably refer to analytics

The role of the blocking buffer is to saturate the remaining sites on the polystyrene wells (ELISA) or nitrocellulose membranes (WB). It is used in a variable percentage from 1 to 5% of a some protein such as BSA, ovalbumin, casein, or powdered milk, or for example fetal calf serum that you are not going to use in cell cultures. It exists also commercial blocking buffers from several providers. For each type of immunoassay, the ideal blocking agent should be found, as well as in what % to use it. I hope it works for you

API certainly has limitations, but you report a major disconnect. Appears the molecular ID may have been on a contaminant. Suggest you ensure pure culture and resubmit.

fyi - the The perfect form of C. kyfir is Kluyveromyces marxianus

CMT is rapid test for detection subclinical cases of mastitis depending on cellular protein(WBC) which detected by using Alkely_Arylsulphonate.

For limits and values depends on degree of mastitis e.g Subclincal,clinical ,chronic ,so degree evaluations according to number of inflamatory cells and kind of aetiology

I am not exactly sure what is happening here. However, unlike many stains, which can be stored for prolonged time periods (in amber viles), Ruthenium Red will only stay good, once mixed, for a couple of days in amber viles. Your problem “may“ be due to stain that is not made fresh.

Wazir ali Hangu, I wonder about the purposefulness of such research with the use of sodium azide and sodium benzoate. After all, when storing food products, such compounds must not be used. However, if you insist on such study, preliminary tests (with different concentrations) will be necessary to determine the effects of these compounds on the components of the walnut milk, here antioxidants.

Yes, harnessing that as an ingredient in formulation of animal fed is possible. You only have to follow the principles of feed formulation including purpose, nutrient requirement of animal, inclusion rates, among others. Very importantly you need to ensure the quality and safety of the that ingredient before its use.

In the case of high inbreeding, which results in a genetic similarity of the clan to a large degree, in this case the difference resulting from the environment, as the quantitative traits are affected by both genetic and environmental factors

To avoid this, out-breeding of the herd must be brought about through mating (crossbreeding)

From the attached image and your content, it seems that issue could be related to either with the sample preparation or the sample buffer itself. The beta actin control indicates that the protein has not been equated in your sample as the beta actin bands are of varied intensity. So protein equation is also concern with the image.

Recheck whether you have used appropriate enzyme (proteinases) inhibitors in your sample buffer and the samples are prepared as per the established protocol.

Best

The usual first step is to centrifuge the milk to remove the fat and precipitate the casein micelles with low pH or precipitating agents. ... If the majority of the product protein associates with either the fat or micelles, this can be used to advantage. https://pubmed.ncbi.nlm.nih.gov/1429861/

Yes, that worked for me

Also check https://pubmed.ncbi.nlm.nih.gov/25925799/

Also check the following good link:

https://www.fao.org/dairy-production-products/resources/glossary/en/?index=f

Dear @Elisabeth E LeClair

thank you for answers. TRIzol is an expensive chemical so I'm looking for a cheaper chemical.

mastitis & foot & mouth diseases well as Rift valley fever are common now due to winter season

Hi Rajae, I'm not aware of any publications specifically looking at transmission of CNO through maternal milk. But there is recent paper regarding administration of CNO to 8-10 week old mice pups that might be of use:

لا اعلم يمكن مراجعة المختصين

5% skimmed milk for a membrane blocking during western blot

Dear Wazir ali Hangu as an inorganic chemist I'm absolutely not a specialist in this discipline. However, I just came across the following potentially useful article which might help you in your analysis:

The good thing about this article is that it is freely available as public full text on RG. Thus you can freely download it as pdf file.

Good luck with your work!

Check https://www.researchgate.net/publication/330556311_Milk_Heat_Treatments_Temperature_Effect_on_Fouling

Hi,

Did you include phosphatase inhibitors in your lysis buffer? In addition, you may enrich the protein by IP and then do western blot. Good luck for your experiments :^)

Joseph -

I noticed in this question thread that among your questions you asked for the "average milk production per day." Would that be for the entire farm/household so that you could obtain a total volume by household farm? If not, if you had that for each cow at the household, you would have that value. If you now wanted to know total milk production for the current population, you might use these data, total previous milk production per household farm, as predictor (independent variable) data, if you have this for every household in the population of interest, and a relatively small current sample of the largest cases, i.e. farm households, plus perhaps a small systematic sample of the rest of the current population, for the response data. Thus you might relatively quickly obtain an estimate (actually a "prediction," not a forecast, since Y is a random variable) of the total milk production for your current population.

I promoted this model-based approach for finding total energy production, for various categories, at the US Energy Information Administration, and realized this could be applied elsewhere, whenever there is a census followed by a sample or samples, say an annual census survey and 12 monthly samples. The systematic sampling part of the sample here would make it possible to check that the regression coefficient estimated here is good enough for the entire population, or to perhaps stratify otherwise (as in the Karmel and Jain reference found in the first paper to follow). I know that systematic sampling is chapter 8 of both the 1st and 3rd editions of Cochran's Sampling Techniques, Wiley, but the estimation here would instead be by a model-based (prediction) method.

So, if the possibility of relatively fast and relatively inexpensive predicted/estimated total milk production would be of interest to you, here is a guide:

https://www.researchgate.net/publication/303496276_When_and_How_to_Use_Cutoff_Sampling_with_Prediction

Here is a way to estimate sample size needs with a model I think may be appropriate for you:

https://www.researchgate.net/publication/261947825_Projected_Variance_for_the_Model-based_Classical_Ratio_Estimator_Estimating_Sample_Size_Requirements

This presentation may have some practical advice:

https://www.researchgate.net/publication/263927238_Cutoff_Sampling_and_Estimation_for_Establishment_Surveys, using prediction,

and this was an invited presentation for mathematical statisticians at the US Energy Information Administration:   

https://www.researchgate.net/publication/319914742_Quasi-Cutoff_Sampling_and_the_Classical_Ratio_Estimator_-_Application_to_Establishment_Surveys_for_Official_Statistics_at_the_US_Energy_Information_Administration_-_Historical_Development, 2017, using prediction. 

I have a project on this topic, as follows:

https://www.researchgate.net/project/Cutoff-and-quasi-cutoff-sampling-with-prediction-for-Official-Statistics, with updates in reverse chronological order.

Cheers - Jim

If either the positive or negative control does not produce the expected result, it indicates that the investigator should reconsider his or her experimental procedure. Zahra Rezaei

Specimen type: β-hydroxybutyrate levels can be measured in serum. Point-of-care devices that can measure β-hydroxybutyrate levels in a single drop of blood within 30 seconds have been developed Sudhakar Goud Karpurapu

Was it native WB or denatured WB? If it was denatured one how did you prepare you sample?

-20 isn't ideal for sample storage, -80 is (for fresh frozen sample). If you are doing a denatured WB and when you say "mix the dye" means "mix the lysate with sample buffer and cook for denaturation", then you can keep the denatured sample in 4 celsius fig for a couple of weeks for no problem.

Also check https://www.tandfonline.com/doi/abs/10.1080/FRI-200061609

Dear Armelle,

I hope the reference below might help.

1. From popular press:

https://www.bbc.com/news/science-environment-46654042)

2. The scientific publication that the popular pressed was based on:

Hello,

Yes, it is better to use BSA.

You should never use milk while detecting phospho protein by Western Blot. Milk contains casein, a phosphoprotein that will bind to anti-phospho antibodies causing non-specific binding and high background. Milk may mask some antigens if they are present in low abundance which would result in faint bands.

For detection of phospho protein by Western Blot you should use 5% BSA in TBST for blocking, primary antibody in 3% BSA in TBST and secondary antibody in 3% BSA in TBST.

Also, you need to add Phosphatase inhibitors like sodium orthovanadate and sodium fluoride in the lysis buffer to prevent dephosphorylation of phosphorylated proteins.

Best Wishes.

check the article below may help you Nadeen Al-Otaiby

I explain these type of issues in the attached paper...have a look

Molecular fluorescence spectroscopy is one of the most sensitive and highly selective spectroscopic methods, which can detect extremely low amounts of chemical substances. In the milk industry, it is used for the determination of vitamins, fatty acids, residual amounts of antibiotics, and the identification of different milk species in dairy products.

Check https://www.theroastedroot.net/how-to-make-walnut-milk/

It has not been answered definitively. Yet, there are obvious effects in postnatal mice that were exposed to lactating mothers that in turn received TAM for certain time periods. It is not certain that TAM is passed through breast milk as it is in order to activate the CreERT2 system in neonates. My guess is that it should work.

Prolonged TAM administration will seize lactation in those mums.

Y

Kindly read this sir.

Unfortunately, I do not have information in this field and I think that you can obtain information in your field of specialization by searching in scientific sites interested in this field. I wish you success and success.

Check https://pubmed.ncbi.nlm.nih.gov/29030127/

Hello

You need to optimize the secondary antibody concentration. May be 1:10000 is too high. You can further dilute the secondary antibody, and you can go upto 1:20000.

Also, use affinity purified secondary antibody.

Good Luck.

You could try to spot-blot your lysate onto a membrane (just 1 µL placed onto the membrane and given a minute to bind) and then develop like a western. This will tell you whether the lysate contains anything that reacts with the primary antibody, irrespective of molecular mass.

Also check https://sfamjournals.onlinelibrary.wiley.com/doi/full/10.1111/j.1365-2672.2010.04905.x

Check https://www.tandfonline.com/doi/full/10.1080/1828051X.2017.1335182

Kindly check the following RG link:

Check https://www.researchgate.net/figure/Chromatographic-patterns-of-skimmed-milk-A-and-whole-casein-B-obtained-from-caprine_fig1_12654825/amp

Also check https://www.intechopen.com/books/molecular-pharmacology/liposome-a-comprehensive-approach-for-researchers

Check https://www.researchgate.net/publication/45187406_Raman_spectroscopic_quantification_of_milk_powder_constituents

Can you explain more for the purpose of help, thank you

COVID-19 greatly affects life in general. As for the animal production sector, it causes great losses due to the inability of farmers to sell their agricultural products such as eggs and milk, as well as its impact on farmers, their work and management of fields and farms, and thus the deterioration of production

Also check https://www.labome.com/method/Protein-Peptide-Tags.html

An identical blocking agent must be included in both blocking and sample dilution buffers. Immune recognition of contaminants in antigen preparations by antibodies present in test sera: e.g. Anti-BSA antibodies present in human sera react with minor contaminants such as BSA in insulin preparations https://www.sciencedirect.com/science/article/pii/S2215016117300122