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Sialic acid content of infant saliva: comparison of breast fed with formula fed infants

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Sialic acid content of infant saliva: comparison of breast fed with formula fed infants

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1. T H Tram,
2. J C Brand Miller,
3. Y McNeil,
4. P McVeagh

1. Human Nutrition Unit, Department of Biochemistry, University of Sydney, NSW 2006, Australia

1. Professor Brand Miller.

Abstract

Sialic acid is found in especially high concentrations in brain gangliosides, and supplementary sialic acid is associated with increased learning behaviour in animals. It was hypothesised that breast fed infants may have higher concentrations of sialic acid in body fluids and tissues because human milk is a rich source of sialylated oligosaccharides, while formulas contain very little. The aim therefore was to compare the sialic acid content of saliva collected from full term infants who were either solely breast fed or formula fed until weaning at 3–5 months of age. Thirty three infants, 18 breast fed and 15 formula fed, were studied at a mean (SD) age of 5 (2) months. The breast fed infants, when compared with formula fed infants, were found to have almost two times more free sialic acid in saliva (mean (SE) 16.0 (2.7) v 8.2 (2.1) mg/l, p < 0.036) and nearly 50% more total sialic acid (47.3 (3.9)v 32.2 (4.4) mg/l, p<0.014). The findings provide a preliminary indication that an exogenous source of sialic acids derived from human milk may contribute to higher concentrations of sialic acid in body fluids. There are important implications for the formulation of human milk substitutes.

• Sialic acid is found in especially high concentrations in the gangliosides of the cerebral cortex of the human brain

• Human milk contains significant amounts of sialic acid but infant formulas contain very little

• In rats, supplementary sialic acid has been shown to increase learning and the concentration of sialic acid in brain gangliosides

• In this study, breast fed infants, compared with formula fed infants, were found to have higher concentrations of sialic acid in saliva

• An exogenous source of sialic acids derived from human milk may contribute to higher concentrations of sialic acid in other body fluids, including brain gangliosides

• sialic acid
• saliva
• human milk
• infant formula

http://dx.doi.org/10.1136/adc.77.4.315

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• sialic acid
• saliva
• human milk
• infant formula

Preterm infants who consume breast milk in the early weeks of life have been shown to have significantly higher developmental scores at 18 months of age and higher IQ scores at 8 years than those who are formula fed, even after controlling for confounders such as socioeconomic status.1-4 The distinction becomes more pronounced as the duration of breast feeding increases.5Human milk contains many factors that may promote the development of the nervous system, including long chain polyunsaturated fatty acids. The results of supplementation, however, have yielded conflicting findings.6 We believe that a sugar called sialic acid, present in human milk and brain gangliosides in large amounts, but virtually absent in some infant formulas,7-9 may be another factor which promotes brain development. In a previous study, we showed that human milk contains between 256 and 879 mg/l sialic acid derived from the human milk oligosaccharides. 10

N-acetylneuraminic acid is the predominant and perhaps only form of sialic acid occurring in humans.11 It is commonly present as a component of oligosaccharide chains of mucins, glycoproteins, and gangliosides. Especially high concentrations of gangliosides (a large group of sialic acid containing glycolipids, composed of sialyloligosaccharide and lipid moieties) are found in the cerebral cortex of the human brain.12 Studies have shown that intraperitoneal and oral administration of freeN-acetylneuraminic acid (= sialic acid) during the early postnatal period in the rat,13 results in a significant increase in the concentration of sialic acid in brain ganglioside and glycoprotein. Addition of gangliosides to cell cultures was also shown to enhance neurite outgrowth and to stimulate regeneration of neurons in the central nervous system.7
12 In rats during the early postnatal period, a decrease in exogenous sialic acid concentration leads to irreversibly impaired learning behaviour. Conversely, supplementary sialic acid improves learning.13

These findings suggest that the normal development of the infant brain may also be dependent on the sufficient production and maintenance of brain gangliosides and therefore dependent on an adequate supply of sialic acid. Mammals can synthesise N-acetylneuraminic acid from simple sugar precursors, however, the human infant liver may not have the full adult capacity for synthesising sialic acids during the early postnatal period.7 This was shown in young rats where the activity of uridine diphosphateN-acetylglucosamine-2-epimerase (responsible for catalysing the first step to the biosynthesis of sialic acids) was low.14

Studies comparing sialic acid concentrations in human infants have not been reported. This study was initiated to determine if there were any differences in the sialic acid content of saliva collected from full term breast fed and formula fed infants. Although preterm infants show greater brain development ex utero than term infants, there is still significant accretion of brain tissue in the first few months of life of term infants. Saliva was chosen as an indicator fluid that is known to have a high concentration of sialic acid (from which the word saliva is derived) and which could be collected non-invasively from healthly young infants. Adult saliva was also studied to allow comparison with previous reports of sialic acid concentration in saliva. The findings provide a preliminary indication that an exogenous source of sialic acids derived from human milk may contribute to higher concentrations of sialic acid in body fluids.

Methods

Thirty three healthy, full term infants, mean (SD) age 5 (2) months were recruited for the study. Eighteen were solely breast fed and 15 were solely formula fed with cows’ milk based preparations until weaning at 3–5 months of age. At the time of study, 10 of the 18 breast fed infants and seven of the 15 formula fed infants received small amounts of other foods in addition to breast milk or formula. Milk contributed the majority of energy intake and, to our knowledge, it was the only significant source of sialic acid in the diet. None of the subjects was receiving antibiotics. Infant saliva (1–1.5 ml) was collected using a sterile pasteur pipette under the supervision of a mothercraft nurse and analysed within one hour of collection. Twenty four adults, mean (SD) age 23 (2) years were studied for comparison. Their saliva was collected by expectorating into a sterile container (2–4 ml) at least two hours after food ingestion. The adult saliva was analysed immediately.

Sailva was centrifuged at 4°C, 11 000 g for 10 minutes and the supernatant removed for analysis. The remaining sediment was washed with saline (7%, pH 7.0) and centrifuged at 4°C, 11 000g for three minutes. The saline supernatant fraction was discarded, and the wash procedure was repeated. The sediment was then resuspended to the original volume with saline (7%, pH 7.0). Sialic acid content was determined by a combined modification of the thiobarbituric acid method of Aminoff15 and the method of Skoza and Mohos.16 The thiobarbituric assay is considered to be more precise and subjected to the least interference than other colorimetric methods for the detection of sialic acid content in saliva. 17

N-acetylneuraminic acid was used as the standard. Aliquots (140 μl) of standard, blank, or sample were added to an equal volume of water. The samples were treated with 70 μl of periodate reagent (25 mM periodic acid in 0.125 N sulphuric acid) and incubated at 37°C for 30 minutes. The reaction was terminated through the addition of 70 μl of sodium arsenite (2% sodium arsenite in 0.22 M hydrochloric acid). Once the yellow colour of liberated iodine had disappeared, 140 μl of thiobarbituric acid (0.1M, pH 9.0) was added and the solution was heated in a boiling water bath for 7.5 minutes then cooled in ice water. Dimethyl sulphoxide (560 μl) was added and the solutions were measured at 549 nm using plastic microcuvettes. In cases where the solution was turbid due to the presence of precipitates, the solution was centrifuged for three minutes before measuring on the spectrophotometer.

Because the method measures free sialic acid only, the salivary supernatant and sediment samples (140 μl) were hydrolysed with 0. 05M sulphuric acid (140 μl) at 80°C for 60 and 70 minutes respectively, to determine the total (free plus released sialic acid) content. The bound sialic acid content was determined by substracting the value of free from total. The final values for sialic acid content of saliva were determined by summing the values obtained for the supernatant and sediment.

All results are expressed as mean (SE) with p<0.05 taken as the level of significance using unpaired Student’s t test and one way analysis of variance.

Results

Saliva collected from breast fed infants was found to have approximately double the amount of free sialic acid and nearly 50% more total sialic acid content than that from formula fed infants (fig1, table 1).

Figure 1

(A) free, (B) bound, and (C) total sialic acid content of infant saliva according to method of feeding.

Table 1

Mean (SE) sialic acid content in saliva of breast fed and formula fed infants.

By comparison, adult concentrations of bound and total sialic acid in saliva (table 2) were higher than that of breast fed and formula fed infants (p<0.05). Free sialic acid concentrations, however, were similar in adults and breast fed infants and both groups were significantly higher than that of formula fed infants (p<0.05). Adult men had more bound and more total sialic acid content than adult women (table 2).

Table 2

Mean (SE) sialic acid content in the saliva of adults

Discussion

To our knowledge, we are the first to report the sialic acid content of infant saliva. Our results show that breast fed infants, when compared with formula fed infants, have almost two times more free sialic acid in saliva (16.0 v 8.2 mg/l respectively) and nearly 50% more total sialic acid (47.3 v 32.2 mg/l). While the mean differences are clearly biologically important, there was considerable overlap between the two groups in all three measures of sialic acid concentration. This suggests that infants may vary markedly in their capacity to synthesise their own sialic acid and/or to absorb it from the diet. Alternatively, infants may show variation in other attributes of their saliva, such as its protein or water content, which ultimately impacts on the final concentration of sialic acid. Clearly, further studies are needed to confirm these provocative findings.

We found no significant difference between the sexes with the mean total sialic acid content being 39.2 and 42.1 mg/l for boys and girls respectively. No correlation was detected between the age of the infant and sialic acid content of their saliva. In fact, the concentrations of free sialic acid in the saliva of breast fed infants, although not formula fed infants, approached that of adults.

It has been shown that the measurement of substances in saliva might be a reliable indicator of plasma and body concentrations of these agents.18 The amount of nitrite in saliva, for example, was found to depend directly on the amount of nitrate and nitrite ingested. 18
19 Dietary supplementation with β carotene (provitamin A) in human subjects results in significantly higher concentrations of β carotene in whole saliva compared with a control group.20 Hence it is highly plausible that the higher value for total sialic acid content in the saliva of breast fed infants reflects the high concentration of sialic acid in breast milk which augments uptake and incorporation into salivary glycoproteins. Like nitrates and nitrites,18 ingested sialic acid is probably absorbed by the upper gastrointestinal tract, concentrated from the plasma and excreted into the saliva by salivary glands. Thus sialic acids in breast milk may supplement a limited capacity for endogenous synthesis of sialic acid in the human infant.

Many researchers have assumed that the gastrointestinal tract of suckling mammals contains significant amounts of sialidase activity that can cleave sialic acid residues from the oligosaccharides in the milk. The extent to which this occurs will influence the amount of sialic acid absorbed in the small intestine and the amount of the exogenous sialic acid available for brain ganglioside synthesis. Dickson and Messer found a direct correlation between intestinal sialidase activity and the sialic acid content in milk of suckling rats and 12 other species.14 Sialidase activity has also been shown to occur in human adult intestinal mucosa, and in gastric and intestinal secretions21 and human milk.22 If substantial amounts of sialic acids derived from human milk are incorporated, then the developmental and intellectual scores of human infants and children may be critically influenced by the nature of feeding in the first few months of life.

It is interesting that in the present study, free sialic acid concentration of breast fed infants was similar to that of adults (16.0v 13.5 mg/l, respectively) while formula fed infants achieved about half this concentration (8.2 mg/l). This observation supports the view that formula fed infants may neither have a sufficient dietary supply of sialic acid, nor are they able to synthesise sufficient amounts endogenously. Previous studies in adults have documented similar concentrations of sialic acid in saliva to those found in the present study,23 with a range from 45 mg/l24 to 63 mg/l25 for total sialic acid.

There are other implications of a higher sialic acid content in saliva of breast fed infants. It has been shown that sialic acid contributes to the viscosity of saliva26 and that the viscosity of saliva and the mucin content are positively correlated.27The main function of mucins is to protect the mucosal surfaces by forming a coating layer thereby acting as a lubricative gel, protecting the oral surfaces against mechanical disruption or abrasion. Thus, it is also possible that the breast fed infant has enhanced physical protective mechanisms as a result of the greater sialic acid content in their saliva.

Further research is needed to determine whether other fluids and tissues such as serum, cerebrospinal fluid, and brain gangliosides of breast fed infants have higher concentrations of sialic acid and, importantly, that this can be related to higher intake from exogenous sources. Any difference may contribute to our understanding of why intelligence scores are higher in breast fed infants. If sialic acid from dietary sources can be shown to influence the development of the human brain, this will have important implications for the formulation of human milk substitutes.

References

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   (1996) Feeding formula without arachidonic acid and docosahexaenoic acid has no effect on preferential looking acuity or recognition memory in healthy full-term infants at 9 months of age. Am J Clin Nutr 64:40–46.

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   (1985) N-acetylneuraminic acid concentrations in human milk oligosaccharides and glycoproteins during lactation. Am J Clin Nutr 41:720–726.

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   (1993) Sialinsaure in muttermilch und sauglingsformelnahrungen. Monatsschrift Kinderheilkunde 141:946–950.

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   (1991) Quantitative determination of complex carbohydrates in bovine milk and milk based infant formulas. J Dairy Sci 74:2860–2871.

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    (1994) The oligosaccharide composition of human milk: temporal and individual variations in monocassharide components. J Pediatr Gastroenterol Nutr 19:371–376.

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    (1995) Biology of the sialic acids. (Plenum Press, New York), pp 7–25.

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    (1982) Sialic acids: chemistry, metabolism and function. (Springer Verlag, New York), pp 196–200.

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    1. Morgan B,
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    (1981) The subcellular localization of administered N-acetylneuraminic acid in the brains of well-fed and protein restricted rats. Br J Nutr 46:231–238.

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    (1978) Intestinal neuraminidase activity of suckling rats and other mammals. Biochem J 170:407–413.

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    (1961) Methods for the quantitative estimation of N-acetylneuraminic acid and their application to hydrolysates of sialomucoids. Biochem J 81:384–392.

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    (1976) Stable thiobarbituric acid chromophore with dimethyl sulphoxide. Biochem J 159:457–462.

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    (1978) The colorimetric analysis of sialic acid in human saliva and bovine salivary mucin. J Dent Res 57:995–1002.

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    (1992) Influence of dietary nitrate on nitrite level of human saliva. Bulletin of Environmental Contamination and Toxicology 48:83–88.

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    (1976) The effect of nitrate intake on nitrite formation in human saliva. Food Chem Toxicol 14:549–552.

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    (1988) Saliva concentrations of some selected proteins and glycoprotein markers in man after supplementary intake of beta-carotene. Int J Vit Nutr Res 58:171–177.

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    (1968) Neuraminidase in human intestinal mucosa. Biochim Biophys Acta 46:91–98.

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    (1976) Demonstration of neuraminidase activity in human blood serum and human milk using a modified radioactively labelled glycoprotein as substrate. Hoppe-Seylers Zeitschrift fur Physiologische Chemie 357:559–566.

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    (1994) Antimicrobial factors, sialic acid, and protein concentration in whole saliva of the elderly. Scan J Dent Res 102:120–125.

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    (1976) The metabolism of O-acyl-N-acylneuraminic acids. Biosynthesis of O-acylated sialic acids in bovine and equine submandibular glands. Eur J Biochem 68:597–610.

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    (1963) Release and breakdown of sialic acid from human salivary gland mucin and its role in the formation of dental plaque. Nature 199:461–467.

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    (1988) Correlation between viscosity and sialic acid content of human saliva. Aichi-Gakuin Dental Science 1:21–27.

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Babies Born with Congenital Cytomegalovirus (CMV)

When a baby is born with cytomegalovirus (CMV) infection, it is called congenital CMV. Most babies with congenital CMV never show signs or have health problems. However, some babies have health problems at birth or that develop later.

Some babies with congenital CMV infection have signs at birth, such as: 

• Rash
• Jaundice (yellowing of the skin or whites of the eyes)
• Microcephaly (small head)
• Low birth weight
• Hepatosplenomegaly (enlarged liver and spleen)
• Seizures
• Retinitis (damaged eye retina)

Some babies with signs of congenital CMV infection at birth can have long-term health problems, such as:

• Hearing loss
• Developmental and motor delay
• Vision loss
• Microcephaly (small head)
• Seizures

Some babies can have hearing loss at birth or can develop it later, even babies who passed the newborn hearing test or didn’t have any other sings at birth.  

CMV is the most common infectious cause of birth defects in the United States. About 1 out of 200 babies is born with congenital CMV.

One out of 5 babies with congenital CMV will have symptoms or long-term health problems, such as hearing loss.

In the most severe cases, CMV can cause pregnancy loss. 

Most people with CMV infection have no symptoms and aren’t aware that they have been infected. If you are pregnant and get infected with CMV, you can pass the virus to your baby during pregnancy. This can happen when you are infected with CMV for the first time or again during pregnancy.

Young children are a common source of CMV

By the age of 5 years, one in three children has been infected with CMV, but usually does not have symptoms. The virus can stay in a child’s body fluids like saliva and urine for months after the infection. People who are around young children a lot are at greater risk of CMV infection.

Parents and childcare providers can lower their risk of getting CMV by reducing contact with saliva (spit) and urine from babies and young children.

• Do not share food, utensils, or cups with a child.
• Wash your hands with soap and water after changing diapers or helping a child to use the toilet.

Congenital CMV infection can be diagnosed by testing a newborn baby’s urine (preferred specimen), saliva, or blood. These specimens must be collected for testing within 2 to 3 weeks after the baby is born to confirm a diagnosis of congenital CMV infection.

For babies with signs of congenital CMV infection at birth, antiviral medications (primarily valganciclovir) might improve hearing and developmental outcomes. Valganciclovir can have serious side effects and has only been studied in babies with signs of congenital CMV infection. There is limited information on the effectiveness of valganciclovir to treat infants with hearing loss alone.

90,000 10 reasons for concern, which should not be worried about

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• Care for newborn: 10 reasons for anxiety, which should not be worried about

NEWBORN CARE: 10 WORRYS NOT TO WORRY ABOUT

Every minute, a newborn baby not only brings you great joy, but also gives you many reasons to worry. We decided to do a little research to understand what new parents should worry about and what can be left unattended.

1. Touching the fontanel – a wide soft area on the baby’s head. It does not matter if you touch the fontanel, it will not harm the child in any way. Do not think that by touching the fontanel, you are directly touching the baby’s brain. So what are you touching anyway? To a dense protective shell. Thanks to the fontanelles, the head of an infant passing through the birth canal is compressed without damaging the brain. By about a year, the fontanels overgrow.
2. Distinct pulse in the fontanelle area . Do not be alarmed if you can literally observe the pulsation of blood in the fontanelle of a newborn – this indicates the normal operation of the baby’s circulatory system. Since the fontanelles are located in places of incomplete fusion of the bones, they have a soft structure, so veins and arteries are visible on them.
3. Blood on the diaper of a newborn girl . During pregnancy, a surge in the mother’s production of the hormone estrogen can stimulate the uterus of the female fetus. During the first weeks of a girl’s life, they often experience so-called. “mini-menstruation”, when bloody discharge from the uterus appears.
4. Small chest cavity for infants . Relax, your child is not sick. According to experts, the sternum consists of three parts. It is possible that the depression you noticed is the lowest part, pointing back. As the child grows, his chest and abdominal muscles will gradually flatten this depression. However, even before that time, layers of eye-pleasing baby fat will hide this completely normal feature of the structure of newborns.
5. Soft watery stools after every feeding . When breastfeeding, newborns often poop after each feed. There is no pathology in this, just breast milk is absorbed very quickly (with artificial feeding, babies poop less often). And the watery consistency of their feces is due to the liquid nutrition of newborns.
6. Constant hiccups . Experts cannot say with complete certainty why babies hiccup so often. Some believe that this is due to a poorly established connection between the brain and the diaphragm – the muscular septum that separates the chest cavity from the abdominal cavity and regulates breathing. Whatever the reason, hiccups are a completely harmless and inalienable attribute of infancy.
7. Tears and screams. The nervous system of newborns is extremely prone to irritation. Newborns are easily frightened. These two reasons are enough to explain why babies shed so many tears. Crying is the only way an infant can express its needs. Simply put, newborns are simply supposed to cry and scream a lot, so you shouldn’t worry too much – your baby is unlikely to be in serious danger.
8. Small pimples on the face . Because of the maternal hormones still in the baby’s body, he often develops acne between 2 weeks and 2 months of age. This rash is completely harmless and requires only the most minor cleansing. No need to wipe it with Clearasil.
9. Swollen breasts in girls… and boys ! The hormones that stimulate mini-menstruation in newborn girls also cause slight breast enlargement in infants of both sexes. Unusual? Yes. Temporarily? Exactly. Should I be worried? In no case.
10. Constant sneezing . Babies have such tiny noses! Even the smallest drop of mucus causes the urge to sneeze. And since your baby has only recently left his cozy watery home in your womb, he can’t escape at least minor inconvenience, which can cause a few sneezes. If sneezing is not accompanied by the release of yellowish thick mucus, indicating a cold, you just need to wish him good health. Soon the sneezing will stop by itself

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9000

Digestion of newborns.

Features of the digestion of infants

Caring for a baby implies not only love, but also an understanding of how the body of a small person works and what processes take place in it. This knowledge is necessary in order to correctly organize the nutrition of the baby, as well as respond to the nuances of his life without panic and surprise.

Dry initial milk formula adapted by Valio Baby 1 NutriValio for feeding children from birth to 6 months
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Follow-up dry milk formula adapted by Valio Baby 2 NutriValio for feeding children from 6 to 12 months
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Dry milk drink “Baby milk” Valio Baby 3 NutriValio for feeding children over 12 months
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While the baby is waiting to be born in the womb, his digestive system is in a state of inactivity. The embryo receives nutrition through the placenta. As soon as a child is born, it becomes necessary to obtain nutrients in the traditional way for a person – through the mouth. The baby’s mind already has a sucking reflex, but it needs to be “launched”. That is why early (ideally in the first half hour of life) attachment of the baby to the mother’s breast is so important. If you give a newborn a nipple first, then the muscles of the oral cavity will remember exactly its shape and the so-called “nipple” sucking will form. In the future, when breastfeeding, the mother will have problems – the child will injure the nipples or completely refuse the breast.

How to improve digestion in infants?

The volume of the stomach of a newborn is only 30 ml. When the baby begins to grow, the capacity of the stomach will also increase – at the age of 1 month it will be about 100 ml, at 3 months – 150 ml, at six months – 200 ml, by 1 year – 250 ml. Mom needs to understand that such a small organ cannot be overfilled. Overfeeding will stretch the walls of the stomach and will contribute to overeating in the future.

Breast milk (and in the early days colostrum, more nutritious, high in protein, very energetically valuable) is an ideal food for a baby. 10 ml of colostrum per feeding in the first days of life will fully satisfy the newborn’s need for food. Together with milk, antibodies to many infectious diseases enter the baby’s body (those that the mother had been ill in her life). Thus, the child receives a kind of vaccination. In addition, milk is the most easily digestible and easily digestible food. This is important, since the process of digestion in newborns is only being formed, and very few enzymes are produced in the stomach to digest food.

One of the important components of the digestive process is saliva. In a newborn, due to the weak development of the salivary glands in the oral cavity, there is little of it. Dry mucous membrane is very susceptible to minor damage and infection. In addition, antibodies (class A immunoglobulins) that protect it are not produced at all in the first month of life. Therefore, babies often develop thrush. With growing up, the production of saliva will increase, but the swallowing reflex will not form immediately, therefore, at 3-4 months, the child will experience frequent salivation. So that saliva does not irritate the skin, it must be wiped periodically.

The stomach of young children is horizontal until the child begins to walk. Food stays in it for an average of 2.5–3 hours. This must be taken into account when organizing intervals in feeding. Artificial formulas take longer to digest than breast milk – an average of 3-4 hours.

The intestines of a newborn are very long, and the muscles of the internal organs are still weak. That is why children in the first 3 months of life often experience constipation. Later the chair is normalized. Usually, children in the first months of life empty their intestines 2-4 times a day. By the end of the first year, 1-2 times a day is considered normal. Be prepared for the fact that in the first two days of life, thick, very dark-colored primordial feces will come out of the baby’s intestines. This is meconium – it accumulated in the baby’s body throughout the entire intrauterine development.

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Author: Reetta Tikanmäki

Palm oil in baby food

Infant milk formulas are made from cow’s milk. However, in terms of fat composition, it differs significantly from that of the mother.