Viagra is a commonly known form of medication Sildenafil. It is used for the treatment of erectile dysfunction that helps men to stimulate their erections by increasing blood flow in the penis. Other than that, it is used for certain heart conditions in most people.

There are many reasons that influence how long the medication lasts and how long it takes before Viagra starts to make noticeable and effective changes. Most often, it takes about 30 minutes before showing results, but it also depends on the following few factors that might affect its time duration.

Diet of an individualThe overall health of the personUsage of other medications (if any)Underlying conditions, etc.

Also, you can get Viagra from Numan for yourself at very affordable prices. Along with that, it has hair loss treatment medications also.

How Viagra Works?

The erection for a man occurs when the nerves of the penis are aroused and stimulated. They help the muscles of erectile tissue that are present around the penis and clitoris, known as the corpus cavernosal to get smooth and relax, in order to allow the adequate amount of blood to flow and cause an erection.

However, with erectile dysfunction, the nerves become incapable of communicating with the brain in a proper way. This leads to improper blood flow into the corpus cavernosa, which helps stimulate an erection.

Viagra works by helping with muscle relaxation and soothes the walls around blood vessels that let blood flow more conveniently into the body parts including the penis. This is how it helps with an erection.

When Viagra Starts Working?

The viagra medication starts to show its effects in about 30 to 60 minutes after taking it orally. It may also take up around 2 hours to work if the above-mentioned factors are influencing the individual.

In addition, to let viagra work normally and a bit sooner, a person still needs to feel comfortable, relaxed, and stress-free along with feeling sexually aroused to get an erection.

For How Long Viagra Works?

On average, the Viagra medication lasts up to two to four hours before its effects start to decrease. However, it can also last for more than four hours depending on various factors such as the dosage, age of the individual, their diet, psychological state, overall health conditions including bodys metabolism, etc.

Moreover, a higher dosage of viagra can also take longer to leave the body. This means that a 25 mg to 50 mg tablet will effect for a couple of hours whereas a 100 mg one may take approximately four times longer to decrease its effects.

A person can also get an erection several times with Viagra in his body depending on how the body metabolizes it. But it may not work again instantly after having sexual intercourse because most likely a person cannot get another erection right after ejaculating. The reason is, the body is not physiologically prepared for it and that is known as the refractory period.

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How Long It Takes For Viagra To Work And How Long It Lasts? - Programming Insider

If you frequently find yourself staring into the fridge, there could be good reason. Humans have five distinct appetites, scientists say, which work in tandem to ensure we get specific amounts of the nutrients our bodies need to work efficiently: protein, carbs, fats, sodium, and calcium.

In studying animal behaviours over the last 30 years, researchers David Raubenheimer and Stephen Simpson authors of Eat Like the Animals: What Nature Teaches Us About Healthy Eating have gone on to make significant discoveries about the human diet.

Their study of a female Cape baboon in the Cape Peninsula of South Africa, published in the journal PLOS One, showed evidence of longer-term nutrient regulation. Over 30 days, they recorded everything the baboon known as Stella consumed.

While the foods varied widely each day, her diet was a strikingly consistent balance of protein to non-protein (fat and carbohydrate) energy across the month. Raubenheimer and Simpson went on to replicate their appetite research in human subjects.

A volunteer group of 10 people stayed in a chalet for a week, eating from a buffet at their leisure for two days. They were split across two groups a high-protein buffet and a low-protein, high-carb, high-fat buffet for two days, returning to the original buffet for the final two.

The results, published in the journal Appetite, revealed that those on the low-protein diet ate more calories and carbs to replenish the missing protein, while those on high-protein diets consumed fewer to compensate for the imbalance.

It is a mistake to think of appetite as a single, powerful drive to eat.

It is a mistake to think of appetite as a single, powerful drive to eat, Raubenheimer and Simpson write in New Scientist. We need separate appetites to keep track of various nutrients, and hence to construct a balanced diet.

Why protein, carbs, fats, sodium, and calcium? Those five have been singled out by evolution for good reasons, the researchers continue. One is that there is a limit to how complex biological systems can get and still operate efficiently. We couldn't have specific appetites for dozens of nutrients.

Another is that these nutrients are needed in very specific quantities. Third, some components, like sodium, were often rare in our ancestral environments and we needed dedicated machinery to seek them out, for example in mineral deposits.

Ultra-processed foods usually contain ingredients that you wouldnt add when cooking at home chemicals, colourings, emulsifiers, sweeteners, stabilisers and preservatives and can be found in all sorts of products, from breads and cereals to ready meals and reconstituted meat products.

They are low in high quality proteins and high in simple sugars and processed carbs, says Dr Aamer Khan, co-founder of the Harley Street Skin Clinic. They may lack certain vital minerals that manufacturers avoid using because of cost.

The more ultra-processed foods we eat, the more calories we need to consume to reach our target quota of protein. And we naturally gravitate towards that target, even when it means consuming excess carbs and fats to reach it.

Charles GullungGetty Images

Ultra-processed foods make us fat, but not because we have strong appetites for the fats and carbs they contain, as is often thought to be the case, Raubenheimer and Simpson write.

Rather, it is because our appetite for protein is stronger than our ability to limit fat and carb intake. So, when protein is diluted by fats and carbs, our appetite for it overwhelms the mechanisms that normally tell us to stop eating fats and carbs.

In much the same way, going overboard on the protein can have its pitfalls, if youre consistently switching your veggies with steak.

A diet biased towards too much protein will not only restrict calorific intake, but also micronutrient and mineral intake, Dr Khan says. This can result in the breakdown of the normal functioning of the healthy body, the slowing of the metabolism and breakdown of the immune system.

Aim for a balance of macronutrients thats protein, carbs and fats at every meal. Protein is used for building and repairing body tissues, says Dr Khan. Carbs give energy; fats give slow release energy and fat-soluble micronutrients and minerals.

Get acquainted with the spice rack at every opportunity. You could even grow your own fresh herbs. Seasoning is important, it gives micronutrients, he says.

And if you tend to gulp down your meal in a few large mouthfuls, try slowing things down. It takes 10 to 15 minutes for your gut to tell your brain you are full, he says.

Above all, channel your inner Stella next time you go food shopping eating a wide variety of different foods will help you reach your macro and micronutrient requirements naturally.

Diets that are well balanced will allow satiation without taking on foods that have a high calorific value, such as carbs and fats, and also allow sufficient intake of the micronutrients and minerals that are essential to the healthy workings of the human body, says Dr Khan.

Eat Like the Animals: What Nature Teaches Us About Healthy Eating by David Raubenheimer and Stephen Simpson is out now (William Collins).

Last updated: 01-06-2020

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Always hungry? Thats because you have 5 different appetites - Netdoctor

Those whove just started the keto diet are always anxious for one important event to occur.

Am I in ketosis?

Sure, most peoples primary goal on a ketogenic diet is to lose weight. A lot of weight.

But the body has to go into fat-burning mode, more accurately known as ketosis, before the weight begins to disappear. Going into ketosis is the first important signal that the diet is starting to work.

Fortunately, it doesnt take a lot of time for most people. And when the body is entering that all-important phase of the keto process, its usually not a secret.

Heres how to tell when youre in ketosis.

How Ketosis Works

Experts could write long dissertations on the biology and physics of ketosis. But well try to explain it in three short paragraphs, so you can understand why its so important.

The body relies on carbohydrates for the energy it needs to function. Those carbs come from the food we eat; theyre broken down into glucose, a type of sugar thats utilized by the body as fuel. Any glucose thats left over is stored in a form called glycogen.

When you start a keto diet youre virtually eliminating carbs as an energy source. That forces the body to look for fuel somewhere else. It starts by using previously-stored glycogen, but what happens when the glycogen runs out?

It starts burning stored body fat to create a secondary energy source known as ketones. And when the body begins producing ketones for energy, it enters the metabolic state called ketosis. (You knew wed get there eventually.) Thats why you lose weight on the keto diet: youre withholding carbs to force the body into ketosis, so it will burn stored fat for energy.

So ketosis signals that youve entered the stage of your diet when youll be able to start shedding pounds.

The Physical Signs of Ketosis

You can tell when youve entered ketosis by using one of two methods. You can test your blood, breath or urine for the presence of ketones. Or you can simply recognize the physical signs of ketosis that your body is exhibiting.

The Keto Flu

The body doesnt normally use ketones for energy, so when it has to make the switchover from glucose things dont go smoothly at first. Ketosis begins in fits and starts, which usually result in a series of very noticeable symptoms. Taken together, the symptoms have been dubbed the keto flu, since they closely resemble the effects of a bad case of influenza (even though theres no virus or bacteria involved).

Within a day or two of starting a rigorous keto diet youre likely to feel tired, even exhausted. It will be difficult or impossible to complete your normal workouts, and you may suffer muscle cramps even if you havent exercised. Youll also have to urinate a lot, because excess ketones bind to water in your body and have to be excreted.

In more serious cases, you can experience nausea and vomiting, difficulty in focusing, constipation or diarrhea, headaches and insomnia. The good news is that the keto flu lasts no longer than a week for most people, and there are ways to short-circuit it.

Many of the symptoms are because frequent urination leads to dehydration and the loss of important minerals. Drinking lots of water or broth, and getting more salt, magnesium and potassium (via diet or supplements) will help quite a bit but dont do it with store-bought electrolyte drinks, because they contain sugar that will knock you right out of ketosis.

Get lots of sleep, slow down on exercise and stick to your diet, and youll get past it and remember, it means youre going into ketosis.

Dry Mouth and Bad Breath

The frequent urination associated with the keto flu often leaves your mouth feeling dry, and youre thirstier than usual, since its easy to become dehydrated. Youll often notice the smell of one ketone, acetone, on your breath as well; most people simply notice this as bad breath thats difficult to get rid of. These are both signs that youre going into ketosis.

Digestive Problems

It probably wont come as a surprise, but your digestive tract has to adapt to functioning in a very different way when you enter ketosis. Thats not just because of the glucose/ketone energy switch, but also because of the drastic micronutrient changes youre making in your diet. That can result in stomach pains, constipation and diarrhea above and beyond the effects of the keto flu. Those will go away, too.

The Start of Ketos Real Effects

When you start to feel less hungry, your focus and concentration improves, and you start to lose some weight, its time to relax. Those are all signs that not only have you entered ketosis, but your body has become fat adapted. In other words, its made the long-term metabolic adjustment to run completely on ketones rather than glucose. This usually occurs after 2-3 weeks on keto.

Just dont be disappointed if the weight loss isnt extreme or only lasts for a little while; at first, youre just losing water weight. The real keto weight loss comes as you continue to stick with your diet.

Measuring Your Ketones

Its unlikely that your body will enter ketosis without making you very much aware of it.

However, some people who only experience a mild case of the keto flu or dont really notice anything different at all will understandably be anxious to find out whats going on with their body.

There are three ways to test for the presence of ketones in your body, and the most reliable is with a blood test. You can certainly ask your doctor or lab to do it, but there are also home testing kits which will analyze your blood. The tests look for a ketone called BHB, and results are measured in millimoles per liter (mmol/L). Once the reading is above 0.5 that means youve started ketosis, and a reading of 1.5-3.0 (or above) means youve reached stable ketosis,

The second-best choice is to use a different meter to check for the presence of acetone in your breath. Those readings are measured in parts per million (PPM) and you should be between 10 and 40 PPM if youre in ketosis. Finally, you can use urine strips which change color to indicate whether youre in ketosis; theyre not as reliable, but theyre less expensive and easier to use.

Even if you rely on physical symptoms to know if youre in ketosis, its still a good idea to have some method of testing your ketones if you plan to stay on the keto diet. It will allow you to make minor adjustments to the diet and remain comfortably in ketosis for the duration.

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HOW TO TELL IF YOU'RE IN KETOSIS - Island Echo

In the fall of 2018, a team of researchers from the Weizmann Institute of Science in Israel published findings that a cocktail of 11 strains of Lactobacillus and Bifidobacterium had minimal immediate impact and no lasting effect on the makeup of the gut microbiome of mice or people. In fact, the probiotic bacteria were not found in any of the fourteen adult participants after supplementation ended.

These recent findings received quite a lot of press and added to growing sentiment among the public that probioticslive microorganisms that are purported to confer benefits on the human hostdont work. Decades of research have shown that most probiotics aren't able to colonize or exert lasting benefits in the human gut. Some critics even suggested that probiotics may not be a promising avenue for treating disease or otherwise improving health and wellness. But we thought: Dont throw the baby out with the bathwaterour work shows that the right probiotic can work in the infant gut. Findings we published in 2017 showed that feeding breastfed babies a probiotic that included a specific strain of Bifidobacterium longum subspecies infantis (B. infantis EVC001) resulted in a 10,000,000-fold average increase in levels of fecal B. infantis. This level persisted for one month after the supplement was consumed, and levels remained elevated for up to one year after treatment.

To understand why the infant gut microbiome changed so drastically over the past century, we sought to understand how the infant gut microbiome forms.

Colonization of the infant gut by B. infantis had protective effects, such as lower levels of potential gut pathogens and fecal endotoxin, an outer membrane component of Gram-negative organisms known to trigger inflammation. We also found that infants given the B. infantis probiotic had reduced intestinal inflammation compared with breastfed infants who did not receive the probiotic. The gut microbiomes of B. infantis supplemented babies harbored fewer antibiotic resistance genesa sign of fewer pathogensand showed less degradation of mucin, a glycoprotein secreted by the intestinal epithelium that protects epithelial cells from direct contact with gut microbes. These data support earlier findings from Mark Underwood and colleagues at the University of California, Davis. In 2013, Underwoods team showed that feeding preterm infants a different strain, B. infantis ATCC15697, resulted in greater increases in fecal Bifidobacterium and reduced levels of potential pathogens compared with infants given a probiotic containing B. lactis.

While the scientific community and the public grappled with repeated findings that probiotic supplements taken by adults are not consistent in effectively colonizing the gut or conferring benefit, we now had convincing evidence that babies gut microbiomes responded incredibly well to specific strains of B. infantis. The question was why.

Hints about the infant microbiome can be found in century-old articles on commensal bacteria in infant feces. W. R. Logan, a clinical pathologist at the Research Laboratory of the Royal College of Physicians in Edinburgh, was the first to report, 100 years ago, that bacteria in fecal smears from breastfed infants were a near monoculture of Bacillus bifidus, which is today known as the genus Bifidobacterium. Fecal smears from formula-fed infants of that time, by contrast, had a diversity of bacteria, with relatively few Bifidobacteriummore similar to the microbial diversity found in todays breastfed infants.

These striking changes in the gut microbiome composition seen over the past century were consistent with our recent finding that the fecal pH in breastfed infants dramatically increased from pH 5.0 to 6.5 within the past 100 years, a change associated with an apparent generational loss of Bifidobacterium and concomitant increase in potential pathogens. The reduction in Bifidobacterium in the gut microbiome of breastfed infants is likely an unintended consequence of medical practices that can save lives but do not support the growth of Bifidobacterium. Such medical practices include treatment with antibiotics to which Bifidobacterium are sensitive; infant formula that doesnt provide the specific food the bacterium requires; and greater numbers of cesarean section deliveries, which bypass the route by which the bacterium is transferred from mother to baby. These medical practices have been implicated in the increased risk for allergic and autoimmune diseases prevalent in resource-rich nations. The reduction in Bifidobacterium and increase in proinflammatory microbes in early infancy is proposed to occur during the critical window of immune system development, and thereby may increase the risk for immune disease later in life.

To understand why the infant gut microbiome changed so drastically over the past century, we sought to understand how this community forms. Infant gut microbiome colonization begins at delivery with exposure to maternal microbesmostly vaginal and fecal microbes for vaginally delivered babies or predominately microbes from the skin, mouth, and surrounding environment in infants born by cesarean delivery. After birth, infants are bombarded by a vast array of microbes found in the environment, including in breast milk, but the species that go on to become durable members of the microbial community are often those transmitted by the infants mothers through physical contact.

Children continue to acquire gut microbiome species from their mothers and others in the community during early life. This stands in contrast to an adults gut microbiome, which is stable and resists change largely because the available space and food is already used by established microbesthe ecological niches are simply occupied in adult guts. Thus, it makes sense that a probiotic has a better chance of persisting in the infant gut, where it faces less competition, and therefore is more likely to have food it can consume and a location where it can grow. A probiotic serves as just one more source of exposure to new bacteria for the infant.

Recognizing this, we began to wonder: In our studies, what ecological niche did B. infantis fill that supported its persistence in infants long after probiotic administration stopped?

Historically, the breastfed infant gut microbiome was a near monoculture of Bifidobacterium (J Pathol Bacteriol, 18:52751, 1913). The formula-fed infant gut microbiome was much more diverse. The breastfed infant gut microbiome and the formula-fed infant gut microbiome are now more similar to the historical formula-fed infant gut microbiome, although modern breastfed infants do have moreBifidobacterium than modern formula-fed infants.

A major factor in determining which bacteria thrive in the gut is the availability of their carbohydrate food sources. Thus, for a probiotic to work in an infant, microorganisms should be selected so that the food source they use most efficiently matches whats availablea food that is present and not already being consumed by other bacteria. We set out to determine what carbohydrates B. infantis consumes in the infant gut.

Naturally, we turned to breast milk, which for millions of years has been the single food that can exclusively nourish and protect babies for the first six months of life. Human milk delivers nutrients as well as non-nutritive, bioactive molecules, including carbohydrates known as human milk oligosaccharides (HMOs). Back in the mid-1900s, Paul Gyrgy, a world-renowned biochemist, nutritionist, and pediatrician from the Hospital of the University of Pennsylvania, and colleagues unknowingly referred to HMOs when they proposed the existence of a bifidus factor, something unique in breast milk that fed Bifidobacterium. While humans cannot digest HMOs, it turns out that Bifidobacterium, especially B. infantis, can. In 2007, our group at UC Davis used mass spectrometrybased tools coupled with microbiology to show that B. infantis gobbles up HMOs as its sole energy source, while other species of Bifidobacterium consume only some HMOs in addition to plant-, animal-, and host-derived carbohydrates.

HMOs are a diverse class of complex carbohydrate molecules synthesized by the mammary gland. With approximately 200 different molecular species, they represent the third most abundant solid component in human milk following lactose and fat. Because HMOs are complex and vary in structure, they are expensive to manufacture. Current infant formulas may contain one or two simple HMO structures, but at a fraction of the concentration found in breast milk. Infant formulas lack the abundance and complexity of HMOs to selectively feed beneficial gut microbes and to bind and neutralize pathogens from the gut.

The bacterial species in the infant gut capable of consuming HMOs can be considered the milk-oriented microbiome (MOM). Although B. infantis appears to be the most efficient consumer of HMOs, other species of Bifidobacterium, in particular, B. breveand B. bifidum, can and do consume some HMOs but also consume plant-, animal-, and host-derived carbohydrates. The Bifidobacterium species that colonize the gut change throughout life in response to available carbohydrates in the host diet. For instance, B. infantis, B. breve, and B. bifidum are MOM bifidobacteria typically found in the stool of exclusively breastfed infants, while B. longum and B. adolescentis, which preferentially consume plant- and animal-derived carbohydrates, are typically found in the stool of adults. Yet there is variation and overlap in the species present at different life stages.

A major factor in determining which bacteria thrive in the gut is the availability of its carbohydrate food source.

Of the MOM bifidobacteria found in the infant gut microbiome, different species may have different implications for the microbiome. For example, when we gave exclusively breastfed infants a supplement with the probiotic B. infantis EVC001, their gut became dominated by the genus Bifidobacteriumupwards of 80 percent relative abundance of the gut microbiomeand potential pathogens made up less than 10 percent of the community. On the other hand, the gut microbiomes of exclusively breastfed infants who were not supplemented with B. infantis EVC001 had much lower levels of Bifidobacterium, with only about 30 percent relative abundance, and potential pathogens constituted about 40 percent of the microbes in their gut, findings that are consistent with previous work from our group and others. This near-monoculture of Bifidobacterium appeared to be driven by B. infantis, which represented about 90 percent of the total Bifidobacterium in infants fed the probiotic. In contrast, B. longum was the predominant gut Bifidobacterium in the control group, followed by B. breve and B.bifidum. These data highlight the vital importance of strain specificity in probiotics, and the combination of the presence of B. infantis and breastfeeding to support a protective gut environment in infants.

To understand how supplementary B. infantis can so successfully outcompete other microbes in the infant gut, we took a deep dive into its feeding strategy. Turns out it is a picky eater, exclusively dining on HMOs, and when HMOs are abundant, B. infantis gobbles them up ravenously. Unlike other MOM bifidobacteria, B. infantis possesses all the genes necessary for the complete, internal degradation of HMOs and preferentially uses HMOs over any other carbohydrate source. Other MOM bifidobacteria such as B. bifidum and B. breve strains display growth capabilities with only a subset of HMOs. B. infantis thus has a competitive advantage when breast milk makes up the entire diet.

A 2008 study from colleagues at UC Davis and their collaborators showed how B. infantis makes quick use of HMOs: with binding proteins to grab HMOs from the gut lumen and transporters to usher them into the cytoplasm, breaking them down into monosaccharides that are then fermented into lactate and the short-chain fatty acid acetate that are secreted from the cell. These end products maintain a lower pH in the intestinal milieu, supporting the transport of these compounds into the intestinal epithelium for use by the host and creating an undesirable environment for potential pathogens. The production of acetate also blocks the infiltration of toxic molecules produced by pathogenic bacteria by enhancing intestinal barrier function and inhibiting pro-inflammatory and apoptotic responses. Recent findings from one in vitro study have shown that the amount of acetate and lactate produced by different bifidobacterial species is dependent on how well they consume the carbohydrates available to them. Hence, feed a carbohydrate-consuming microbe its preferred carbohydrate, and it has greater potential to produce more of its protective end-products.

Another reason why B. infantis outcompetes other bifidobacterial strains in the gut of breastfed infants is that all of its HMO digestion happens inside the bacterial cell. B. bifidum, on the other hand, digests HMOs externally. This extracellular digestion liberates simple carbohydrates and may cross-feed other species of Bifidobacterium, but also cross-feeds and thus opens an ecological niche for other, perhaps less beneficial microbes. Cross-feeding among microbes diversifies the gut microbiome, which is considered to be generally beneficial in adults.

But is there an advantage to having a near monoculture of Bifidobacterium in infants? By asking this question, our focus turned to immune development.

Human milk oligosaccharides (HMOs) are complex carbohydrates that microbial species of the milk-oriented microbiome (MOM) can use as a food source. Bifidobacterium infantis encodes many proteins that specifically bind and transport all types of HMOs into its cell and digest them internally. Other Bifidobacterium species digest only some HMOs and some do so externally. Digestion of HMOs by MOM Bifidobacterium results in the production of lactate and the short chain fatty acid acetate, that are secreted into the gut lumen. These molecules lower the pH in the intestinal milieu, which improves their transport into the epithelium for use by the host and creates an undesirable environment for potential pathogens such as E. coli.

B. infantis preferentially consumes all HMO species over any other carbohydrate source.

B. bifidum eats only a subset of HMOs.

The decline of Bifidobacterium in infant gut microbiomes and the associated dysregulation of the microbial community, with more numerous potential pathogens, has been suggested as one possible contributor to the increased incidence of autoimmune diseases that plague residents of resource-rich nations. Conversely, observational studies have shown beneficial immune effects of having a fecal microbiome dominated by Bifidobacterium. In two studies in Bangladeshi infants and young children, fecal B. infantis and Bifidobacterium abundances at two months of age were strongly correlated with improved vaccine responses at six months and two years old compared with infants not colonized by B. infantis or with low relative abundances of Bifidobacterium.

Additionally, bifidobacteria are less likely than other microbes, especially potential pathogens, to carry and share antimicrobial resistance genes, which can lead to a higher risk of antibiotic-resistant infections. In an observational study of Bangladeshi and Swedish infants, a dominance of intestinal Bifidobacterium was associated with a significant reduction in both the number and the abundance of antibiotic resistance genes. Moreover, compared with matched-control breastfed infants, supplementation with B. infantis EVC001 led to a reduction of antibiotic resistance genes by 90 percent, a drop largely driven by a reduction in levels of Escherichia, Clostridium, and Staphylococcuspotentially pathogenic bacteria that play a major role in the evolution and dissemination of antibiotic resistance genes.

In an effort to restore the Bifidobacterium-dominated infant gut microbiome that was typical of breastfed babies 100 years ago, we decided to conduct a randomized, controlled trial using the B. infantis EVC001 probiotic. Given that not all B. infantis strains consume all HMOs efficiently, we selected B. infantis EVC001 because we knew this strain had the full cassette of genes needed to fully digest all HMOs. Healthy, full-term, breastfed infants were randomized to consume B. infantis EVC001 for 21 consecutive days starting on day 7 postnatal or to not receive the probiotic.

A PROBIOTIC THAT STICKS: Scanning electron micrographs of infant fecal samples show a large increase in the number of Bifidobacterium microbes in those treated with a probiotic called EVC001 (right) compared with controls (left).

Compared with breastfed control infants who did not receive the probiotic, supplementation resulted in a 10,000,000-fold average increase in levels of fecal B. infantis and increased fecal Bifidobacterium by 79 percent during the supplementation period, and this was still true at one month post supplementation. This means Bifidobacterium colonization persisted without the continuation of probiotic supplementation. Additionally, colonization of B. infantis persisted until one year of age if infants were continuing to consume any breast milk and were not exposed to antibiotics. Importantly, the supplemented infants exhibited an 80 percent reduction in potential gut pathogens belonging to the families Enterobacteriaceae and Clostridiaceae and reduced fecal endotoxin. Additionally, we saw a 2-fold increase in fecal lactate and acetate and a 10-fold decrease in fecal pH. The supplemented infants gut microbiomes and biochemistry resembled norms observed a century ago.

We also identified some clues about the consequences of the gut microbiomes modernization. Breastfed infants with low fecal Bifidobacterium had excreted 10-fold more HMOs in their stool throughout the two-month study period than infants supplemented with B. infantis EVC001, indicating that HMOsthe third most abundant component in breast milkwere going to waste. We also foundthat infants with low fecal Bifidobacterium had several-fold higher levels of fecalproinflammatory cytokines compared with infants whose gut microbiomes were dominated by Bifidobacterium post supplementation with B. infantis EVC001.

Taken together, these data demonstrate that this particular strain of B. infantis, provided as a probiotic to breastfed infants, dramatically colonized the infant gut microbiome during and after supplementation, and beneficially remodeled the microbial, biochemical, and immunological environment in the infant gut. Many infants around the world never acquire B. infantis, but the combination of breastfeeding and probiotic supplementation with this bacterium seems to lead to a nourishing and protective gut environment.

Many infants around the world never acquire B. infantis, but the combination of breastfeeding and probiotic supplementation with this bacterium seems to lead to a nourishing and protective gut environment.

Our findings also support the hypothesis that the ineffectiveness of some probiotics in adults is due in part to the fact that they are introducing a new species to an established community with few ecological niches still open. Probiotics may not work in infants when there is a mismatch between the carbohydrate needs of the probiotic and the availability of highly specific carbohydrates such as HMOs in breast milk. Because B. infantisefficiently consumes almost all HMOs found in breast milk, it is likely to find an open ecological niche and then outcompete other microbes, especially proinflammatory pathogens.

Many scientists are working to understand what the infant gut microbiome really means for health across the lifespan. Meanwhile, we are turning our attention to other questions: How do colonization patterns of Bifidobacterium differ in infant populations around the world from infancy to weaning? And what solid foods support a healthy gut and immune system? Working with funding from the National Institutes of Health, we are now conducting a study designed to understand how the carbohydrate structures of complementary foods influence microbial function that will support a healthy gut microbiome and immune system development in late infancy and early toddlerhood. The ultimate goal is to identify specific carbohydrate structures in the diet that selectively feed beneficial gut microbes in children during the critical window of immune development for lifelong health.

Jennifer Smilowitzis the associate director of the Human Studies Research Program at the Foods for Health Institute and a research scientist in the Department of Food Science and Technology at the University of California, Davis.Diana Hazard Taftis a postdoctoral research fellow in David Millss lab in the Department of Food Science and Technology and a member of the Foods for Health Institute at UC Davis.

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The Infant Gut Microbiome and Probiotics that Work - The Scientist

Abstract

Maize is a cultigen of global economic importance, but when it first became a staple grain in the Americas, was unknown and contested. Here, we report direct isotopic dietary evidence from 52 radiocarbon-dated human skeletons from two remarkably well-preserved rock-shelter contexts in the Maya Mountains of Belize spanning the past 10,000 years. Individuals dating before ~4700 calendar years before present (cal B.P.) show no clear evidence for the consumption of maize. Evidence for substantial maize consumption (~30% of total diet) appears in some individuals between 4700 and 4000 cal B.P. Isotopic evidence after 4000 cal B.P. indicates that maize became a persistently used staple grain comparable in dietary significance to later maize agriculturalists in the region (>70% of total diet). These data provide the earliest definitive evidence for maize as a staple grain in the Americas.

Maize is a staple grain of global dietary importance (1), shaping ecosystems, landscapes, cultures, fire regimes, and biodiversity. However, the early adoption of this crop as a staple (here defined as >25% of total diet) is unknown because of the dearth of archeological sites in the Americas containing early skeletal assemblages needed to directly assess its dietary contribution. Here, we use stable isotope evidence from an unparalleled time series of human skeletal samples, excavated from two rock shelters in Belize spanning the past 10,000 years, to reconstruct the dietary importance of maize through time. We demonstrate that maize became a significant dietary staple for some individuals starting between 4700 and 4000 calendar years before present (cal B.P.) and a persistently used staple grain by 4000 cal B.P. (>70% of total diet). The demographic and environmental impact of this transition had far-reaching consequences for the people of Mesoamerica that have now been amplified on a global scale, particularly in attaining food security in the context of increasingly volatile global climate change.

Maize was domesticated from teosinte (Zea mays spp. parviglumus), a wild grass growing in the lower reaches of the Balsas River Valley of southwestern Mexico (Fig. 1), and molecular clock estimates suggest that it was domesticated in the Early Holocene (~9000 cal B.P.) (2). Microfossils (phytoliths and starch) confirm the use of maize in the Balsas by ~8700 cal B.P. (3). Microfossil data also support widespread dispersal of this important domesticate through the lowland neotropics by at least ~7000 cal B.P. (4). However, the initial dietary significance of maize and the process and timing of its adoption as a staple carbohydrate in the human diet remain unclear. Two rock shelters in Belize are the only known contexts in the Americas containing human skeletal material deposited persistently over the past 10,000 years and during the transition to maize-based food production. Bone preservation in these dry rock shelters provides an unparalleled opportunity to study dietary change associated with the introduction of maize into the region and its subsequent increasing economic and dietary importance across the New World.

Paleoenvironmental sequences are shown as green triangles as follows: (A) San Andres, Tabasco (36); (B) SOC05-2 (19); (C) Lake Puerto Arturo (11); (D) Peten Lakes region (57); (E) Cob Swamp (38); (F) Pulltrowser Swamp (38); and (G) Lake Yojoa (58). Archeological sites with early maize are shown as red dots: (1) Ocampo Caves (59), (2) Xihuatoxtla (3), (3) El Riego (60), (4) San Marcos (60), (5) Coxcatln (60), (6) Guil Naquitz (5), and (7) Caye Coco (25). Map was produced in ArcGIS 10.4, with all subsequent layout and design performed in Photoshop CC 14.2.

Early maize cobs (~6250 cal B.P.) from Guil Naquitz in the Mexican highlands are small and have only two seed rows (5). Ancient DNA data from the earliest Tehuacn Valley cobs (53004950 cal B.P.) indicate a mixture of modern maize-type alleles, controlling stalk and inflorescence architecture, and ancestral teosinte alleles, controlling ear shattering and starch biosynthesis (6, 7). In addition, paleoethnobotanical, paleoecological, and paleogenomic data from South America suggest that maize was dispersed by foraging populations, possibly in a semidomesticated state, as early as ~7000 cal B.P. (8). These observations have led some to speculate that domesticated maize was initially of little dietary importance but instead spread because of the sugar content of stalk juice, enhancing its social significance as a fermented beverage (9). Selection for larger, multirow cobs by ~4300 cal B.P. (10), along with paleoecological studies indicating increased burning, forest clearance, and erosion associated with maize pollen (11), suggests the increasing importance of maize as a staple crop, but even its dietary significance at that time remains elusive.

Stable isotopes from radiocarbon (14C)dated archeological bone collagen (13Ccollagen and 15Ncollagen) combined with bone apatite (13Capatite) provide a powerful measure of the dietary significance of maize in the Americas. 13Ccollagen is a proxy for the primary sources of protein in an individuals diet. Variation results primarily from the isotopic composition of animals and the plants they consume and how the primary producer plants metabolize carbon during photosynthesis. The lowland tropics of Mesoamerica are dominated by plants using the C3 (Calvin-Benson) photosynthetic pathway with distinctively negative 13C values [~26.5 per mil ()] (12). Maize was the primary plant domesticated in the lowland neotropics, and it uses the C4 (Hatch-Slack) photosynthetic pathway with distinctively more positive isotopic values (~9.3) (13). Isotopically, the increasing dietary importance of this domesticate should be evident with more positive 13C ratios. The consumption of marine resources can complicate the interpretation of 13C data by mimicking the consumption of C4 plants if they are not evaluated in conjunction with nitrogen isotope data. Differences in nitrogen isotope values (15N) are sensitive to the trophic position of plants and animals consumed and are used to distinguish between terrestrial and aquatic food consumption (14). Herbivores are enriched by 3 to 7 relative to the plants eaten, and carnivores are enriched by 3 to 5 compared to the animals eaten. Marine mammals and fish are generally enriched by 5 to 10 relative to the terrestrial mammals. However, fish from coral reef ecosystems have high 13Ccollagen and low 15Ncollagen values that overlap isotopically with maize (15). In addition, 13Ccollagen is strongly biased to the protein component of the diet, but 13Capatite generally reflects the whole diet (carbohydrates, lipids, and proteins) (16). Maize kernels contain 8 to 11% protein and 73% carbohydrate, but maize stalk juice is largely composed of carbohydrates and sugar. Therefore, the consumption of fermented beverages from carbohydrate-rich and protein-poor sugary stalk juice (e.g., maize beer or chicha) would be more likely to be visible in 13Capatite and not in 13Ccollagen (17).

Human skeletal material in the Americas is relatively rare before ~3000 cal B.P., and the decomposition or contamination of bone collagen and apatite in the humid lowland neotropics has largely limited stable carbon and nitrogen isotope studies of skeletal material dating before this time. Until our work, the handful of samples available in the lowland neotropics were too poorly preserved to yield reliable results or were from earlier studies that analyzed unpurified collagen that was not directly radiocarbon-dated to verify age (18). One of these studies on the Pacific coast of southern Mexico provides a potentially important time series of samples spanning the last 4500 years (17). The two earliest samples in that study (from contexts thought to be 4500 and 4000 years old) have 13Ccollagen and 15Ncollagen, consistent with significant maize consumption, a result compatible with evidence for the intensification of maize farming in the region during this time (19). However, individuals from contexts thought to date to between 3400 and 2900 cal B.P. showed less dietary reliance on maize. In addition, resources from an adjacent estuary (e.g., shrimp and crabs) were shown to overlap isotopically with maize and confounded interpretation of these data (17). The absence of a clear maize signature in individuals dating between 3400 and 2900 cal B.P. led Blake and colleagues (9) to hypothesize that maize was being consumed as a beverage fermented from stalk juice, but this has not been tested directly with 13Capatite measurements. Overall, chronological and interpretive difficulties in the dataset obscure the dietary importance of maize through time, and the earliest skeletal samples in this time series do not predate the introduction of maize to the region (~6500 cal B.P.) (19).

Stable isotope dietary studies in the Mesoamerican neotropics are most extensive in the Maya lowlands, where emphasis has been placed on variability during the Preclassic (30001700 cal B.P.) (20) and Classic (17001000 cal B.P.) (21, 22) Periods. These studies show strong and increasing reliance on maize after 3000 cal B.P. (23), age- and sex-dependent differences in maize consumption (24), greater access to meat and maize in elite populations (24), and regional variability in the access to maize and marine foods (22). All of the skeletal materials analyzed in these studies postdate the introduction of maize to the region by ~6500 cal B.P. (25) and its intensification after 4300 cal B.P. (10, 11), so dietary variability before and during the transition to maize-based food production is unknown. To evaluate dietary change through time, we obtained a stable isotope dietary data transect from 52 directly radiocarbon-dated human skeletons spanning the past 10,000 years from two remarkably well-preserved rock-shelter sites, Mayahak Cab Pek (MHCP) and Saki Tzul (ST), located in the Maya Mountains of Belize.

MHCP and ST are rock shelters located in an interior valley of the Maya Mountains in the Bladen Nature Reserve (BNR) [162928.98 N, 885437.42 W; 430 m above sea level (masl); Fig. 1), a protected wilderness area where there has been minimal modern human disturbance of archeological sites. The Classic Period Maya settlement of Ek Xux sits in a valley between these two rock shelters, and the much larger Classic Period center of Muklebal Tzul is located only 3 km away. Tikal, Caracol, and 63 other major Classic Period population centers with dated inscriptions occur within 200 km of these rock shelters, 13 with comparative baseline 13Ccollagen, 15Ncollagen, and 13Capatite data from agriculturalists consuming maize (22).

The terrain in the Maya Mountains is rugged (~400 to 1000 masl), and MHCP and ST are positioned above active floodplains at ~430 masl along the Bladen Branch of the Monkey River and the Ek Xux Creek, respectively. Neotropical broadleaf forest predominates in the region and provides a range of edible tree fruits, nuts, and seeds. Palms are sources of fiber, thatch, and food (26), most notably the Cohune (Attalea cohune) palm found in high-density stands that produce a rich source of nuts/oils, fronds used for roofing, and large palm hearts historically used as a famine food (27). Most of the edible plants in this environment use the C3 photosynthetic pathway with the exception of Amaranthaceae, with pollen common in Pleistocene-age sediments (28). Trace amounts of Amaranthaceae pollen has been used as a disturbance indicator in Holocene paleoecological sequences (11). The most notable large prey animals in this region are three deer species (white-tailed, Odocoileus virginianus and brocket deer Mazama americana and Mazama pandora), tapir (Tapirus bairdii), and two peccary species (Tayassu pecari and Pecari tajacu). These animals are largely herbivores and can exploit the interface of the C3 tropical forest and areas of disturbance (29). The Monkey River provides freshwater mollusks (Pachychilus spp.), crabs, and small fish. Overall, protein and carbohydrate availability are dispersed, relatively low density, and seasonally modulated in neotropical forests and would not support concentrated human populations without agriculture (26).

Within this forested environment, MHCP is formed by an east-facing 20-m-high limestone outcrop that creates a 26-m-wide and 6-m-deep rock shelter. Dry sediments and limited root activity inside the dripline (~160 m2) have favored the preservation of bone and carbonized plant materials (30). One 2.5 mby2.5 m excavation trench (figs. S1A and S2) in the center of the rock shelter revealed a ~2.8-m sequence of cultural midden and mortuary deposits. The lowest stratigraphic units (G to K; fig. S2) are organic-rich (silt to silty loam) and contain debris from the limestone cliff outcrop, igneous flaked stone tools of local origin (choppers and hammer stones), large chert bifaces (Lowe points) (30), and animal, riverine shellfish (Pachychilus spp.), and human remains. These deposits do not contain pottery and date between 12,000 and 6000 cal B.P. The upper portion of the sequence (units A to D) is composed of alternating layers of organic-rich rocky sediment and a dense Pachychilus spp. midden. These deposits date after ~3000 cal B.P. and contain pottery fragments, flaked stone chert, and igneous tools and the remains of mammals, birds, and reptiles. Stratigraphic units E and F are transitional between the ceramic and preceramic units and date between 6000 and 3000 cal B.P. and contain Pachychilus midden deposits.

ST formed below another sheer limestone cliff face that is located 1.4 km to the northeast of MHCP across the Bladen River. The shelter sits 70 m above the river, and it is less than 300 m away from the Classic Period Maya center of Ek Xux. It is larger (145 m long and 8 to 15 m wide) than MHCP and has ~1700 m2 of dry sediments inside the dripline. Multiple excavation units (figs. S1B and S3) reveal a parallel sequence to MHCP spanning the past 12,500 years. Artifact density is high in the upper ceramic-bearing strata containing high concentrations of animal bone, burned wood, and disarticulated human remains. Two dense Pachychilus lenses (>70% shell) occur just below these mixed deposits. The preceramic deposits dating to the Middle Holocene are dominated by dark midden sediments and high concentrations of Pachychilus shells and contain stone tools, bone, carbonized plant material, and human skeletal material. The Early Holocene sediments change to a relatively compact light gray silt and contain lower, but consistent, concentrations of Pachychilis shells, stone tools, carbonized plant material, and human burials.

Human remains at both rock shelters reflect similar burial practices and include both primary and secondary burials (section S1). Isolated elements were recovered from many stratigraphic levels, and while some are the result of intrusive disturbance into earlier contexts, others are intentional deposits of individual skeletal elements or multiple elements. Although a wide range of burial practices are reflected in this long mortuary transect, burials in varying degrees of flexure were the most common and are found in all time periods. In total, 63 inhumations have been excavated from both rock shelters, not including isolated remains. Here, we report data for 52 individuals represented by 32 adults, 4 juveniles, 13 infants, and 3 indeterminate age (see dataset S1 and section S1 for details).

We obtained 13Ccollagen, 15Ncollagen, and 13Capatite data and accelerator mass spectrometer (AMS) radiocarbon (14C) dates for 30 individuals from MHCP and 22 individuals from ST (Fig. 2 and dataset S2; also see Materials and Methods and section S2 for methodological details and our standards for quality control). Both males (n = 13) and females (n = 12) are represented in the sample of adults; however, the sex for 27 individuals could not be determined because they were incomplete skeletons or too young. Infants under the age of 3 were included in the analysis, but we subtracted 2 from 15Ncollagen values to offset well-documented enrichment associated with nursing (31). 13Ccollagen in infants largely reflects the mothers diet, but some enrichment can occur. However, we did not correct for 13Ccollagen because it is highly variable and less than 1. Juveniles over the age of 3 rarely show 15Ncollagen or 13Ccollagen enrichment associated with nursing (31). The most ancient individuals in this dataset come from the lower cultural strata at MHCP and date to between 9600 and 8600 cal B.P. (dataset S1). Two other individuals date earlier than 6000 cal B.P., and the remainder was persistently buried at these locations until ~1000 cal B.P., with a possible hiatus between 3200 and 2700 cal B.P.

(A) Pre-maize diet (96004700 cal B.P.), (B) transitional maize diet (47004000 cal B.P.), and (C) staple maize diet (40001000 cal B.P.). For more details on the skeletal sample in this study, see section S1 and age model parameters in section S2. The radiocarbon plot produced in OxCal 4.2 with subsequent layout and design was performed in Illustrator CC 17.1.

In this study, we group skeletal remains into three chronological categories based on 13Ccollagen, 13Capatite, and 13Cenamel data that are presented below: pre-maize diet (96004700 cal B.P.), transitional maize diet (47004000 cal B.P.), and staple maize diet (40001000 cal B.P.). Statistical significance between temporal periods was determined using a one-way analysis of variance (ANOVA) and Tukeys post hoc test with significant differences observed among 13Ccollagen (n = 47; ANOVA; F2,44 = 290.0, P < 0.001, 2 = 0.929, = 0) and 13Capatite (n = 34; ANOVA; F2,39 = 66.2, P < 0.001, 2 = 0.772, = 0). All post hoc results were corroborated using nonoverlapping bias-corrected and accelerated (BCa) confidence intervals (95% CI) with 1000 bootstrap replicates and verified with permutation test. We evaluated changes in 13C using a linear mixing model to estimate the dietary contribution of maize (%) based on its distinctive C4 photosynthetic pathway (32). We also appraise diet based on a carbon isotope model (33) and a multivariate isotope model (22), both calibrated with controlled feeding studies. We also use a large sample of Classic Period (17501000 cal B.P.) individuals with demonstrated staple maize diets as a comparative baseline to track changes in maize consumption through time (22).

13Ccollagen values (n = 14) in the earliest individuals (older than 4700 cal B.P.) range between 21.6 and 20.3 (mean = 20.8; SD = 0.3) and indicate minimal or no C4 plant consumption (Fig. 3A). When combined with nitrogen isotopes (n = 14; 15Ncollagen, 6.4 to 9.9), these measurements are consistent with a population consuming C3 plants and terrestrial animals from lowland tropical environments. The 15Ncollagen values are, on average, ~3 to 6 higher than published values for herbivores in the region (O. virginianus and M. Americana) (34). Carbon isotope enrichment in these early humans indicates that the consumption of marine foods is not evident. We cannot rule out small dietary contributions of aquatic resources such as small gastropods (Pachychilus spp.), which are abundant throughout the midden sequence and overlap isotopically with C3 plants (34). Other domesticates such as squash (Cucurbita spp.) or manioc (Manihot esculenta) also cannot be ruled out because they also use the C3 photosynthetic pathway and overlap isotopically with wild plant foods found in neotropical forests. 13Ccollagen values may not be sensitive to C4 plant consumption, especially if only carbohydrate-rich portions of the plant were consumed (e.g., sugary stalk juices) (4, 9).

(22) (A) 13Ccollagen versus 15Ncollagen. (B) 13Ccollagen versus 13Capatite plotted against dietary regression lines from experimental feeding studies (33). (C) Function 1 versus function 2 discriminant analysis plotted against data from experimental feeding studies (35). Data analysis in R, with subsequent layout and design performed in Illustrator CC 17.1.

We measured 13Capatite to evaluate total diet (proteins, lipids, and carbohydrates) and to determine whether C4 plants were a substantial part of the total diet. 13Capatite values (n = 15) range between 15.4 and 11.9 (mean = 13.6; SD = 1.0), and comparable results were found in four paired 13Cenamel samples (dataset S1). A simple linear mixing model (32) of 13Capatite indicates a C4 contribution to total diet between 0 and 21% (average = 10.5%; SD = 6.1). Therefore, we cannot rule out minimal consumption of C4 plants (e.g., Amaranthaceae or maize), particularly in the case of three individuals (MHCP.14.1.6 and MHCP.17.1.8, and ST.18.11.9) with values between 16 and 21% C4 dietary contribution coming from C4 carbohydrate-rich source(s). However, 13Capatite and 13Ccollagen plotted against C3 and C4 protein regression lines and calibrated with published archeological and experimental data (33) show that these individuals still cluster on the C3 protein line, indicating a close to pure C3 diet (combined protein and carbohydrates; Fig. 3B), but with small-scale variations in resource consumption. Discriminant function analysis (F1 and F2) in a multivariate model including 13Capatite, 13Ccollagen, and 15Ncollagen (35) shows clustering of samples within the 100% C3 diet space (Fig. 3C).

Carbon isotopic enrichment consistent with an increase in C4 consumption occurs in some individuals after 4700 cal B.P., but diets are highly varied (n = 10, mean = 18.3; SD = 2.2) for ~700 years. The age profile of this sample (including seven individuals younger than 3 years of age) contributes to this variability, but the nursing signature shows that some mothers were consuming substantial amounts of C4 plants. 13Ccollagen values are significantly different from the pre-maize dietary values, indicating that individuals were consuming a protein-rich C4 plant, like maize (8 to 11% protein by kernel weight), or consuming animals eating C4 plants (P < 0.01). There is no statistical difference between 15Ncollagen isotopes (7.0 to 9.9; P = 0.576), and these values do not suggest a major change in the source of protein (e.g., aquatic foods). Therefore, the enrichment in the 13Ccollagen in some individuals reflects a significant increase in the dietary importance of C4 plants or animals consuming C4 plants.

13Capatite values are consistent with an increase in the dietary importance of C4 plant consumption, most certainly maize, ranging between 14.9 and 6.6 (n = 10; mean = 11.2; SD = 2.3; P = 0.04). This translates to C4 plant consumption averaging 25.8% (SD = 14.1) of total diet based on a simple linear mixing model (32). Bivariate plots of 13Capatite and 13Ccollagen show separation of some individuals in this group from the pre-maize diet along the C3 protein line, indicating an increase in C4 protein and carbohydrate consumption consistent with increased maize consumption (Fig. 3B) (33). Discriminant functions (F1 and F2) in the multivariate model show the majority of transitional farmers in the 30% C4 diet space (65% C3 protein; Fig. 3C). Three individuals dating within this interval have isotopic values that are indistinguishable from pre-maize diets and highlight the variability during this transitional period. Permutation tests show low retest reliability in the mean comparisons; however, the observed difference and simulation absolute-threshold differences suggest that transitional maize diets are more enriched than the simulated baselines (figs. S4 to S6). This largely results from high variability and small sample size of this group.

13Ccollagen values for individuals dating after 4000 cal B.P. (n = 23) range between 13.5 and 8.2 and overlap with values for staple maize diets during the Classic Maya Period (Fig. 3A). Comparisons indicate that these values are significantly different from pre- and transitional maize diets (P < 0.001). 15Ncollagen values (6.1 to 10.2) are comparable to earlier populations. There is no evidence for the consumption of marine fish from pelagic environments (low 15Ncollagen isotope values), but we cannot rule out the possibility of some coral reef fish in the diet obtained via trade (22). However, the bones of these animals are largely absent in associated middens, even though we have used techniques designed to recover small bone samples (e.g., 200-m mesh sieves). Overall, the patterns indicate the consumption of forest-dwelling herbivores and heavy reliance on maize.

13Capatite values range between 11.9 and 4.4 (mean = 6.7; SD = 1.9) and are consistent with 21 to 68% (mean = 53.7%; SD = 11.7) of total dietary carbon coming from a C4 source with significant differences between pre-maize and transitional diets (P < 0.001). Bivariate plots of 13Capatite and 13Ccollagen fall on or near the C4/marine protein line and are consistent with staple maize diets from the southern Maya lowlands (Fig. 3B) (22). Discriminant function analysis shows all of the post4000 cal B.P. samples overlapping with individuals from the southern Maya lowlands in the 70% C4 range and with >50% of dietary protein coming from C4 plants or animals eating C4 plants (Fig. 3C). Permutation results support all post hoc differences tested with staple maize diets (figs. S4 to S6).

MHCP and ST are the only archeological deposits in the Americas that contain human skeletal material deposited persistently over the past 10,000 years and the only sites in the lowland neotropics that span the transition to maize-based food production. Preservation of bone organics in these dry rock shelters provides an unparalleled opportunity to study dietary change associated with the introduction of maize into the region and its subsequent development as a food staple of increasing economic and dietary importance. Edible plants of economic value using the C4 photosynthetic pathway are rare in the neotropical lowlands, and this makes stable carbon and nitrogen isotopic analysis a powerful tool for tracking the dietary importance of maize through time.

Isotopic evidence in individuals dating between 9600 and 4700 cal B.P. is consistent with dietary dependence on plants and animals from a C3-dominated neotropical forest. 13Ccollagen values indicate that minimal C4 plant consumption and 15Ncollagen are consistent with the consumption of forest-dwelling herbivores (e.g., white-tailed and brocket deer). We cannot completely rule out minimal consumption of a carbohydrate-rich C4 plant source (e.g., Amaranthaceae or maize) based on slightly elevated 13Capatite values in some individuals based on a linear mixing model (32). Evidence for maize cultivation in the Maya lowlands first appears at ~6500 cal B.P. (25) at about the same time that it appears along the Pacific Coast of Mexico (SOC05-2) (19) and Mexicos Gulf Coast lowlands (36), and it is possible that after this time, maize was adopted in the region for its sugary stalk. However, when 13Ccollagen, 13Capatite, and 15Ncollagen are considered together and evaluated against data from controlled feeding studies (33, 35), all individuals dating before 4700 cal B.P. are consistent with 100% consumption of C3 plants and animals consuming C3 plants. These data are also consistent with the early dietary importance of neotropical trees, particularly cohune palm (27), and tubers in early foraging economies in the neotropical lowlands. Use of these rock shelters until ~5600 cal B.P. appears to be persistent but episodic, suggesting low-density populations exploiting a resource-poor neotropical forest (26).

The first evidence for a significant dietary contribution of maize as a staple (defined here as >25% of total diet) occurs between 4700 and 4000 cal B.P. During this transitional interval, there is a clear increase in both 13Ccollagen and 13Capatite in most individuals consistent with increased maize consumption. However, three individuals overlap isotopically with pre-maize diets, indicating variable reliance on maize during this period. Higher 13Ccollagen and 13Capatite indicate that by this time, maize protein and carbohydrates (kernels) were being consumed. We cannot rule out the consumption of glucose-rich stalk juice, and it is likely that both stalks and kernels were used in beverage preparations. Our data partially overlap isotopically with an early population from South America (Pacopampa, Peru) (37) with higher 13Capatite values that possibly indicate maize beer consumption, but those 13Ccollagen values are not as 13C-enriched as the MHCP and ST individuals. The isotopic data during this transitional interval are inconsistent with the hypothesis that maize was solely used for its sugary stalk in a beverage (17). Overall, our multivariate model of 15Ncollagen, 13Ccollagen, and 13Capatite indicates that maize provided about 30% of total diet (Fig. 3C). These data are consistent with evidence for increased forest burning and clearing, maize cultivation, and erosion across the Maya lowlands, suggesting a greater commitment to maize farming between 4500 and 4000 cal B.P. (11, 38). Increases in maize productivity may have resulted from the development and/or the introduction of new landraces (10) or technological innovation (e.g., nixtamalization) (39). Increasing dietary dependence on maize as a staple grain in this region also coincides with the widespread adoption of maize throughout Mesoamerica (Fig. 4) and its diffusion into the United States by ~4100 cal B.P. (40).

The earliest radiocarbon dates associated with microbotanical evidence for maize in the Balsas region (Xihuatoxtla) (3), Mexicos Gulf Coast (San Andrs) (36), and the Maya region (SOC05-2, Caye Coco, Lake Yojoa, Cob Swamp, and Lake Puerto Arturo) (9, 10, 12, 13, 16) are also shown. Summed probability distributions and dietary phases produced in OxCal 4.2 with subsequent layout and design were performed in Illustrator CC 17.1.

Individuals in our sample dating after 4000 cal B.P. overlap isotopically with a large dataset from the Classic Period (17501000 cal B.P.) Maya populations (22). In this larger dataset, elites were more enriched isotopically than commoners, and our samples overlap more with the commoner population, indicating equal amounts of C3 and C4 foods and relatively high in C4 protein sources like maize or maize-fed animals. A persistent contribution of reef fish is largely discounted because of the interior position of most sites, and this would certainly be the case for MHCP and ST located at least a 2-day walk from the coast and where marine food residues have not been identified in associated archeofaunal assemblages. These data point to a strong commitment to maize-based food production combined with the continued exploitation of foods from the neotropical forest that may have involved more sophisticated forest management systems (4). The dietary dependence on maize is consistent with greater investments in surplus agricultural production, and the resulting deforestation, soil degradation, and erosion as populations increased in size and aggregation (41).

Our results suggest that maize-based food production and dietary dependency on maize came to form the economic basis for these developments but did not stimulate them immediately. Increasing dietary dependence on maize between 4700 and 4000 cal B.P. precedes archeological evidence for the earliest pottery-making agricultural villages in the Maya region by over 1000 years (ca. 31003000 cal B.P.) (23, 25). It remains unclear whether the transition to maize-based food production and the associated dietary changes resulted from the influx of a new population into the Maya lowlands. Linguistic data suggest that Proto-Maya diversified out of the western Guatemalan Highlands likely around 4200 cal B.P. as Huastecan speakers moved northeastward and then eventually settled along the Gulf Coast of Mexico, followed shortly after by diversification of Yucatec speakers into the Maya lowlands (42). Proto-Mayan contained words for maize planting, harvesting, and processing of maize, minimally indicating some cultivation of the domesticate (43). Given uncertainties associated with glottochronology and its partial reliance on archeological data, it is plausible that increases in the dietary reliance of maize co-occurred with language dispersal and population movements. Before the arrival of Yucatec speakers, the lowlands were inhabited by unknown non-Maya populations who we show were consuming increasing amounts of maize as new people and new varieties of maize were moving into the region. It has been suggested that they may have spoken a language typologically similar to lower Central American Xinkan, Lenkan, or Tol based on limited phonemic evidence (43). The term for maize in Xinkan derives from a very early, possibly Proto-Mayan root, suggesting that Xinkan foragers adopted the word after contact with early Maya farmers in the Guatemalan Highlands (44). The linguistic evidence also hints at the complex demographic history in the region. We now know that the earliest colonists in North and South America were genetically distinct from modern Maya populations (45), but it remains unclear how long these ancestral colonizing populations persisted in the region before being replaced or admixed. Coinciding with the transition to agriculture and the emergence of societal complexity leading to the Classic Period, the dietary shifts evident in our dataset therefore provide tantalizing evidence for the origins of Maya people in the lowland neotropics.

MHCP and ST are highly significant because they are the only archeological sites in the Americas with a relatively continuous diachronic sample of human skeletal material spanning the past 10,000 years. Therefore, the stable isotope analysis of these directly radiocarbon-dated individuals provides an unprecedented view of dietary changes during the transition from foraging to farming in the Americas. In the lowland neotropics, these data complement genetic and paleobotanical evidence for the domestication and spread of maize after ~9000 cal B.P. Paleoecological records in the Maya region suggest that the initial introduction of maize, in its earliest form, occurred between ~6500 and 5500 cal B.P. (25), but maize had little dietary impact as a staple before 4700 cal B.P. Consumption increased after this time, and it became a persistent dietary staple by 4000 cal B.P. Comparable isotopic studies indicate the adoption of maize as a staple elsewhere in Mesoamerica by ~3000 cal B.P. (9), in South America no earlier than 3500 cal B.P. (46), and in North America later than 2500 cal B.P. (47, 48). Ultimately, this transition contributed to the expansion and aggregation of populations, increases in social inequality, and major environmental transformations.

Permits for field research in Belize and permissions to export and conduct direct dating and isotopic analyses of ancient human remains were issued by the Institute of Archaeology, National Institute of Culture and History, Belize, with additional permits to conduct fieldwork in the BNR issued by the Forest Department (FD), Belize. They are the legal entities responsible for permitting research. Research permit applications were also formally reviewed and supported by our local collaborator, the Yaaxch Conservation Trust (Yaaxch), a Belizean conservation nongovernmental organization strongly committed to preservation of environmental diversity and heritage. Yaaxch co-manages the BNR with the Belize FD. Yaaxch is largely staffed and administered by members of local communities, some of whom are park rangers working alongside our archeological team conducting fieldwork with other members of local communities. From 2016 to 2020, the results of our field and laboratory studies were presented to members of local communities through events organized by Yaaxch and as a condition of archeological permits and in mutual support of our ongoing collaboration. The results of both field and laboratory studies have been presented annually (2014 and 20162019) at the Belize Archaeology Symposium, a public conference attended by members of many diverse communities in Belize.

Carbon and nitrogen isotope ratios were measured on extracted and purified bone collagen or the amino acids that comprise bone collagen. Bone collagen was extracted and purified using a modified Longin method with ultrafiltration (49). Samples (200 to 400 mg) were demineralized for 24 to 36 hours in 0.5 N HCl at 5C, followed by a brief (<1 hour) alkali bath in 0.1 N NaOH at room temperature to remove humates. The residue was rinsed to neutrality in multiple changes of H2O (18.2 megohm per cm) and then gelatinized for 12 hours at 60C in 0.01 N HCl. The resulting gelatin was lyophilized and weighed to determine percent yield as a first evaluation of the degree of bone collagen preservation. Rehydrated gelatin solution was pipetted into precleaned Centriprep ultrafilters (retaining >30-kDa molecular weight gelatin) and centrifuged three times for 20 min, diluted with H2O (18.2 megohm per cm), and centrifuged three more times for 20 min to desalt the solution.

In most instances, bone collagen samples were too poorly preserved for ultrafiltration, and amino acids that comprise bone collagen were extracted from bone samples and pretreated using a modified XAD process (50). The sample gelatin was hydrolyzed in 2 ml of 6 N HCl for 24 hours at 110C. Supelco ENVI-Chrom SPE (solid-phase extraction; Sigma-Aldrich) columns with 0.45-m polyvinylidene difluoride filters were equilibrated with 50 ml of 6 N HCl, and the washings were discarded. Two milliliters of collagen hydrolyzed as HCl was pipetted onto the SPE column and driven with an additional 10 ml of 6 N HCl dropwise with a syringe into a 20-mm culture tube. The hydrolyzate was lastly dried into a viscous syrup by passing UHP (ultra-high purity) N2 gas over the sample heated at 50C for ~12 hours. The isotope ratios of extracted amino acids may not be directly comparable to collagen ratios, but the differences are minor (50).

Carbon and nitrogen concentrations and stable isotope ratios of the collagen or amino acid samples were measured at the Yale Analytical and Stable Isotope Center with a Costech elemental analyzer (ECS 4010) and Thermo DELTAPlus analyzer. Sample quality was evaluated by % crude gelatin yield, %C, %N, and C/N ratios. C/N ratios ranging between 3 and 3.5 indicated good collagen or amino acid preservation (49).

Bone collagen or individual amino acids were directly dated at the Pennsylvania State University (PSU) AMS radiocarbon dating facility. Samples (Ultrafiltration, ~2.1 mg; XAD, ~3.5 mg) were combusted for 3 hours at 900C in vacuum-sealed quartz tubes with CuO and Ag wires. Sample CO2 was reduced to graphite at 550C using H2 and an Fe catalyst, with reaction water drawn off with Mg(ClO4)2 (49).

Graphite samples were pressed into targets in Al boats and loaded on a target wheel, and 14C measurements were made on a modified National Electronics Corporation (NEC) compact spectrometer with a 0.5-MV accelerator (NEC 1.5SDH-1). The 14C ages were corrected for mass-dependent fractionation, with 13C values measured on the AMS (51) and compared with samples of Pleistocene whale bone (backgrounds, 48,000 14C B.P.), late Holocene bison bone (~1850 14C B.P.), late 1800s CE cow bone, and OX-2 oxalic acid standards for normalization. All calibrated 14C ages, probability distributions, and phase boundaries (section S2 and dataset S2) were computed using OxCal version 4.3 (52) with the IntCal13 northern hemisphere curve (53).

The preparation for carbonate analysis in bone was conducted in the Human Paleoecology and Isotope Geochemistry Laboratory at the PSU using a modified version of procedures outlined in (54). A 2.5 2.5 cm fragment of bone shaft was cleaned using a mechanical drill so that the outer layer of the bone cortex and all trabecular bone were removed from the sample. Using an agate mortar and pestle, bone samples were ground into a fine powder. For each sample, 50 to 100 mg of bone powder were reacted in 2 ml of 2% bleach (NaOCl) for 24 hours at room temperature with vented capped vials and then rinsed three times in H2O (18.2 megohm per cm) or until the sample reached a neutral pH. Following the bleach treatment, samples were reacted in 2 ml of 0.1 M acetic acid (CH3COOH) for 24 hours at room temperature with vented capped vials (55) and then rinsed again to a neutral pH with water (18.2 megohm per cm). The samples were then dried overnight at 60C and subsequently analyzed at the Center for Stable Isotopes, University of New Mexico by continuous-flow isotope ratio mass spectrometry using a GasBench device coupled to a Thermo Fisher Scientific Delta V Plus isotope ratio mass spectrometer. The results are reported using the delta notation measured against VPDB (Vienna Pee Dee Belemnite). Reproducibility was better than 0.1 for both 13C based on repeats of a laboratory standard (Carrara Marble). The laboratory standard is calibrated versus National Institute of Standards and Technology (NBS)-19, for which the 13C is 1.95.

Statistical analyses were performed in SPSS 25.0 (IBM SPSS, Chicago, IL, USA) and R software. The P value threshold for statistical significance was 0.05 for all tests. A priori power analysis (1) and effect size test (2) were performed for the primary computational results. Individual burials were categorized into three groups: pre-maize diets, transitional maize diets, and staple maize diets. Normality was evaluated using the Shapiro-Wilk test, and homogeneity of variance was assessed using Levenes test. A one-way ANOVA was applied to compare differences followed by Tukeys post hoc test and Dunnetts T3 post hoc test in the cases of heterogeneity of variance. Statistical differences were corroborated via nonoverlapping BCa confidence intervals (95% CI) with 1000 bootstrap replicates. We evaluated the post hoc results using permutation tests with 1000 randomized iterations and resampling in R (figs. S4 to S6). Plots were created in R using ggplot2. Descriptive statistics are available in dataset S1.

Analysis of carbonate (CO3) in biogenic hydroxyapatite [Ca10(PO4)6(OH)2] was first tested for diagenetic alteration using Fourier transform infrared (FTIR) spectroscopy at the Materials Characterization Laboratory at the PSU using a Bruker Vertex 70v FTIR spectrometer with a DiaMax attenuated total reflection accessory. Each spectrum was the result of 100 scans for mid-range IR (4000 to 400 cm1) with a spectral resolution of 4 cm1. Corrected baselines for the spectra were calculated by adding the heights of the absorptions and then dividing by the height of the minimum between them (56). A new background was created for each sample run. To determine the degree of apatite recrystallization, the crystallinity index or infrared splitting factor was calculated using the height of the absorption bands at 603 and 565 cm1 divided by the height of the valley between them at ~595 cm1. All but two measured sample spectra had a crystallinity index less than 3.8, which indicated a well-preserved biogenic signal, and remained in this study for analysis. While the ratio of the absorption peak height at 1415 cm1 (CO3) and 1035 cm1 (PO4) indicates degraded carbonate material, all sample spectra are consistent with expected C/P ratios observed in archeological bone (56), and none of the spectra indicate CaCO3 contaminant absorption at 710 cm1 (figs. S7 to S10).

This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial license, which permits use, distribution, and reproduction in any medium, so long as the resultant use is not for commercial advantage and provided the original work is properly cited.

S. H. Ambrose, L. Norr, in Prehistoric Human Bone: Archaeology at the Molecular Level, J. B. Lambert, G. Grupe, Eds. (Springer Berlin Heidelberg, 1993), pp. 137.

M. Blake, Maize for the Gods: Unearthing the 9,000-Year History of Corn (University of California Press, 2015).

R. H. Tykot, N. J. van der Merwe, N. Hammond, Stable Isotope Analysis of Bone Collagen, Bone Apatite, and Tooth Enamel in the Reconstruction of Human Diet, in Archaeological Chemistry (American Chemical Society, 1996), vol. 625 of ACS Symposium Series, pp. 355365.

D. R. Piperno, D. M. Pearsall, The Origins of Agriculture in the Lowland Neotropics (Academic Press, 1998).

L. E. Wright, Diet, Health, and Status Among the Pasin Maya: A Reappraisal of the Collapse (Vanderbilt Univ. Press, 2006).

R. H. Tykot, R. L. Burger, N. J. van der Merwe, The Importance of Maize in Initial Period and Early Horizon Peru, in Histories of Maize: Multidisciplinary Approaches to the Prehistory, Linguistics, Biogeography, Domestication, and Evolution of Maize, J. Staller, R. Tykot, B. Benz, Eds. (Academic Press, 2006), pp. 187197.

J. Staller, M. Carrasco, Pre-Columbian Foodways (Springer Science, 2010).

L. Campbell, Mayan loan words in Xinca, in The Mayan Languages (Routledge Language Family Series, 2017), pp. 62111.

T. Kaufman, Aspects of the lexicon of proto-Mayan and its earliest descendants, in The Mayan Languages (Routledge Language Family Series, 2017), pp. 62111.

M. A. Katzenberg, Prehistoric Maize in Southern Ontario: Contributions from Stable Isotope Studies, in Histories of Maize: Multidisciplinary Approaches to the Prehistory, Linguistics, Biogeography, Domestication, and Evolution of Maize (Academic Press, 2006), pp. 263270.

Acknowledgments: We thank the Belize Institute of Archaeology (permits) and the staff of Yaaxch Conservation Trust for logistical support. We thank the following members of the Human Paleoecology and Isotope Geochemistry Laboratory for their assistance processing AMS 14C radiocarbon and stable isotope samples: L. Eccles, M. Davis, L. Crouthamel, X. Moreno, and L. Simmins. AMS 14C radiocarbon dates from this project were analyzed at the PSU AMS 14C facility and the W. M. Keck Carbon Cycle Accelerator Mass Spectrometry Laboratory. We thank T. TJ Zimudzi at the PSU Materials Characterization Laboratory for his guidance during the FTIR analysis and N.-V. Atudorei at the UNM Center for Stable Isotopes for the 13C analysis of carbonates. Thanks to H. Neff, L. Kistler, R. Rosenswig, H. Thakar, R. Tykot, D. Piperno, J. Capriles, S. Plog, S. Newsome, J. Kennett, and two anonymous reviewers for guidance and valuable comments that helped improve the manuscript. Funding: The work was funded by the Alphawood Foundation (20142019; K.M.P.) and NSF (SBE1632061, K.M.P.; SBE-1632144, D.J.K. and B.J.C.). General laboratory support at the PSU from NSF Archaeometry program BCS-1460369 to D.J.K. and B.J.C. Author contributions: D.J.K., K.M.P., B.J.C., and M.R. designed research; D.J.K., K.M.P., M.R., W.R.T., R.J.G., B.J.C., G.M.B., E.M., E.J.K., T.K.H., L.O., E.E.R., E.C.H., A.A., C. Merriman, C. Meredith, and H.J.H.E. collected data; D.J.K., K.M.P., R.J.G., B.J.C., J.J.A., S.M.G., and T.K.H. analyzed data; and D.J.K. and K.M.P. wrote the paper, with contributions from all authors. Competing interests: The authors declare that they have no competing interests. Data and materials availability: All data needed to evaluate the conclusions in the paper are present in the paper and/or the Supplementary Materials. Detailed fieldwork reports including excavation forms, photographs, and artifact inventories are permanently stored at the Institute of Archaeology (Belmopan, Belize) and are available from the authors upon request.

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Early isotopic evidence for maize as a staple grain in the Americas - Science Advances

Finding out how to lose weight shouldn't be so complicated. Once you start eating more mindfully and do some exercise, you should naturally start to lose belly fat without having to put yourself through the horrors of intermittent fasting or the keto diet although both have a lot of fans, and a certain amount of scientific evidence to support them. One way to help the weight loss process is to speed up your metabolism and to do that, we've collected 3 easy-to-follow tips that you can implement today.

An improved rate of metabolism can help you keep weight off, and generally turn you into a healthier and leaner individual. As usual, there's no instant fix to be had here, but being fit and having a faster metabolism quickly become a virtuous circle.

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According to research cited by Healthline, "proteincauses the largest rise in TEF (thermic effect of food). It increases your metabolic rate by 1530%, compared to 510% for carbs and 03% for fats." As Healthline explains, the "thermic effect of food caused by the extra calories required to digest, absorb and process the nutrients in your meal."

Since digesting protein takes more effort from your body, by eating more of it you will basically work out using your metabolic system (we might be exaggerating here). Protein is also essential for muscle repair and recovery, so if you are actively working out, it is recommended to take between 1.6-2 grams of protein per body kilogram per day.

You should source protein from a variety of food stuff like lean meat, nuts, green veg and eggs. Supplementing protein is also popular among athletes: protein powder shakes are probably the most convenient way to get your protein fix on the go but you can also have protein bars or jerky as well as post workout snack.

(Image credit: Sage)

Caffeine found in coffee and teas can boost metabolism significantly. Not everyone reacts to caffeine the same way and it is also recommended not to drink too many cups of coffee/caffeinated beverages in a day. Green tea has a lower caffeine content so combining coffee and green tea consumption can have better results.

Drinking more water can also improve metabolism: combine increased water consumption with more fibrous food for the best results.

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Stuck in a weight loss rut? Here's how to speed up your metabolism to blast belly fat - T3

It goes without saying that the COVID-19 pandemic has caused inconceivable human suffering and will worsen gender-based inequalities.

As economic activity grinds to a standstill, women, who were already disadvantaged when it came to accessing decent work, will suffer the most.

In Assam, the unemployment rate is likely to increase and settle in the range between 14% and 25% from the current level of 8% and poverty ratio is expected to rise to 50.8% for the state in the worst possible scenario with a 15% fall in income. The figure is estimated to be 54% for rural Assam.

Estimates from the National Flood Commission of the Ministry of Water Resources show that 31.60 lakh hectares of area are vulnerable to floods in Assam which amounts to 9.4% of the total flood-prone area in the country and affects one-third of the population in the state. The reproductive healthcare and nutritional security of women and children are therefore severely challenged with the additional burden of a possible escalation in poverty in the wake of the outbreak of the coronavirus and impeding flood during monsoons.

The National Family Health Survey (NFHS-4) shows that approximately 36% of women in Assam had a Body mass index (BMI) that was below normal and 8% were obese. Anaemia was prevalent in 72% of pregnant women and 69% of women who were not pregnant. In addition, teenage pregnancy (15-19 years age group) was at 61.4% and 13.6% of them were already mothers.

Also read: COVID-19 Has Pushed Indias Already Suffering Tea Plantation Workers into Deeper Crisis

The low BMI and anaemic conditions amongst women in Assam contribute towards reinforcing structural deficiency in child health. There are 36% children in the state with stunted growth (NFHS-4). Furthermore, adequate breastfeeding and supplementary diet for children has decreased to 49% in 2015 (NFHS-4) compared to 60.1% in 20015 (NFHS-3) in the state.

Only 8.9% of children during the first two years of their lives received an adequate diet. It is therefore not difficult to understand why the incidence of infant mortality rate(IMR) and nutritional deficiency continues to be high in Assam. The low nutritional intake during the first two years of early childhood is also likely to lead to cognitive impairments and further impact growth and development.

Tribal women are seen sowing rice crops in an agricultural field in the outskirt of Nagaon Town in Assam. Photo: Flickr/Diganta Talukdar

Womens diet is an important indicator of a childs health. The NFHS-4 data shows that the protein content of diet in Assam is around 7% in rural and 10% in urban areas. The per capita per diem intake of protein in rural Assam is 54.4 gm and for urban areas is 58.8 gm. The average calorie intake per capita per diem in rural Assam is 2120 and 2176 in urban areas and the urban calorie intake per capita is above the recommended calorie intake for poverty level (2100 calorie per day per person) in India but for rural areas, it is below the poverty level (2400 calorie per day per person).

The NSSOs 66th round data indicates that 59% rural and 52% of urban households in Assam face a calorie deficit (<2700 calorie intake per consumer unit per day).

As per the PLFS 2017-18 data, the unemployment rate for females is 13.6% and 7.2% for males in the state. The labour force participation rate (LFPR) for females is 12.7 and the same for males is 80.3% which means a higher proportion of females stay away from active economic work in the state.

The worker population ratio (WPR) is 10.8% for females and 74.1% for males. The average wage for a male worker is Rs 17,375.5 and for a female worker is Rs 12, 891.9 with a male-female wage differential of Rs 4,484. These add to womens household vulnerabilities with access to nutritional sufficiency.

Also read: In Assam, the Healthcare Apparatus for Women Needs a Complete Overhaul

Household vulnerabilities are compounded further given that 75% of the total workers in the state are either self-employed or comprises of casual workers and of the 25% in regular employment, 49% are non-regular.

Therefore access to income and work uncertainty for households is acute during the current pandemic. With a halt to economic activities, poverty and deprivation will escalate. The AHDR, 2014 showed that over the past one and a half-decade, the state lost about one-third of the potential aggregate human development due to the prevailing inequalities underlying achievements in education, health and income dimensions. The loss due to inequality was highest in income dimension (44%) followed by health (32%) and education (9%).

The COVID-19 pandemic is likely to affect an estimated 67 lakh peoples livelihood with myriad vulnerabilities and contribute to worsening of household wellbeing with a spillover effect on nutritional deficiency. This is likely to worsen the RCH which may contribute towards reversing the current improvement in MMR and IMR. These will have far reaching consequence on overall human development in the state in years to come. The road map to revamp state economy in fighting Covid-19 cannot ignore this aspect.

Dr Saswati Choudhury is senior faculty at Omeo Kumar Das Institute of Social Change and Development in Guwahati.

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Healthcare and Nutrition for Women in Assam Was Already Inadequate. Then Came COVID-19 - The Wire

As many offices and other large buildings begin to reopen following months of being unoccupied during the state's stay-at-home order, New Jersey American Water is reminding building owners, operators and managers to take part in proper flushing procedures to keep its water clean and safe.

Matt Csik, director of water quality and environmental compliance for New Jersey American Water, said when water sits in the internal pipes of a building without use for a long time, lead from those pipes can flake into the water. Sitting water can also allow dangerous bacteria such as legionella to grow and enter the water.

Building managers need to flush the water that's been sitting in those internal pipes so fresher water comes through.

To do that, Csik said they need to run each cold-water tap in the building for at least two minutes. Once someone notices a temperature change or a chlorine smell, that's a good assurance that they're pooling fresh water from the system all the way to the tap.

Csik said if building owners don't properly flush all the pipes, they could expose people to levels of lead that has entered the water from the internal pipes of the building. It also can create a problem where bacteria has grown in the pipes. That can create water droplets that people can inhale and become sick.

Ideally, if a building owner knows the tap that is the furthest from where the water enters the building from the pipes, start with that tap and flush it until the chlorine smell or the temperature change indicates fresh water is flowing through the system. Then work back to where the water enters the building, flushing each and every tap.

He said it's also a good idea to flush showers, letting them run for a few minutes, and to flush toilets at least two times.

Another thing people need to be careful about is ice makers and refrigerators that have water dispensers. Csik said building managers need to make sure they make follow the owners manuals to make sure they are properly flushed before people use them.

Csik said this kind of problem with pipes can happen under any circumstances if a building is sitting for a long period of time without use. But with so many buildings shut for a long time due to the COVID-19 pandemic, so many businesses are now reopening months later. It's even more important now that proper flushing protocols be taken.

"We're just really reminding our customers to make sure they're protecting anybody who could consume the water from those buildings," said Csik.

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As NJ buildings reopen, they need to flush pipes to keep water safe - wobm.com

Government spending plans to help businesses suffering in the novel coronavirus pandemic have again come under fire over the amounts earmarked for paperwork as well as a lack of transparency over the programs.

The Constitution requires all government expenditures to first gain Diet approval.

However, a reserve fund of 10 trillion yen ($92 billion) planned in the second supplementary budget is recognized as a measure for unforeseen deficiencies in the budget, meaning that it can be spent as the Cabinet wishes without gaining prior approval of the Diet.

Opposition lawmakers said they would not simply stand by and allow that second supplementary budget to pass unchallenged.

It would be an act of suicide for the Diet to give the government carte blanche to spend 10 trillion yen, Jun Azumi, the Diet Affairs Committee chairman of the Constitutional Democratic Party of Japan, said.

Diet deliberations on the second supplementary budget will begin next week.

Another target of criticism during June 3 Diet deliberations was the central governments program to encourage consumers to spend more to help businesses that have been particularly hard-hit by stay-home requests to reduce the risk of COVID-19 infections.

The Go To Campaign involves various measures, such as issuing coupons that can be used at tourist destinations or restaurants and providing subsidies to local shopping malls to organize events to attract consumers.

The total cost of the Go To Campaign is 1.7 trillion yen. But 309.5 billion yen, or about 18 percent of the total, has been set aside to handle the paperwork and other administrative matters for the measures.

Tourism minister Kazuyoshi Akaba explained that between 13 and 23 percent of the amounts for previous tourism support measures were earmarked for administrative matters.

However, the past tourism support programs were smaller in scale and designed for specific areas, such as prefectures damaged by natural disasters.

The Go To Campaign will cover the entire nation.

The Ministry of Economy, Trade and Industry has been accepting applications since May 26 from the private sector to handle administrative procedures for the Go To Campaign.

The deadline for submitting applications is June 8, but the ministry remains vague on how the 309.5 billion yen will be used.

Opposition lawmakers said the administrative expenses should be sharply reduced so that more money can reach companies in the tourism sector facing serious difficulties in the coronavirus pandemic.

Satoshi Arai of the CDP suggested that the central government funds be distributed to prefectural governments that would provide the money to local tourism associations and chambers of commerce, which would have a better grasp of where support is most needed.

An economy ministry official explained why such a large amount is needed to handle administrative matters.

The official said reports are required for how the funds are used, call centers must be set up to handle inquiries from businesses and consumers interested in the programs, and publicity is needed for the campaign.

The official also cited personnel expenses and the creation of new computer systems for the program.

However, the ministry has not provided a detailed breakdown for each of those expense areas.

More questions were raised in the Diet about the Service Design Engineering Council, which has been commissioned by the government to handle paperwork for the subsidy program for small businesses.

The council is being paid 76.9 billion yen for the work, but it has turned around and outsourced the job for 74.9 billion yen to advertising giant Dentsu Inc., one of the companies that was involved in establishing the council in 2016.

It now turns out that the council has never released an annual financial statement as is required by law.

Opposition lawmakers who tried to find out what the council actually did said visits to its Tokyo office led to nothing because no one was available to answer questions.

Fueling the criticism from the opposition was the economy ministrys admission that it was considering paying the council about 85 billion yen to handle additional paperwork for measures included in the second supplementary budget.

The 10 trillion-yen reserve fund in the extra budget will come on top of the 1.5 trillion yen set aside for the reserve fund in the first supplementary budget passed in late April.

So far, the largest total for the reserve fund over the course of a year was the approximately 2 trillion yen set aside in fiscal 2011, when the nation was reeling from the Great East Japan Earthquake, tsunami and nuclear disaster.

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Cabinet accused of wasting more funds on virus countermeasures : The Asahi Shimbun - Asahi Shimbun