When is it necessary to take a stool test for opportunistic flora? Interpretation of the analysis for dysbacteriosis in adults and children

Intestinal infection is a common cause of digestive upset in children. To quickly cure your child, you need to get tested and identify the true cause of the intestinal disorder. Laboratory diagnosis of intestinal infections includes stool analysis for an opportunistic group of microorganisms and analysis for pathogenic flora (analysis for the disgroup and typhoparatyphoid group of bacteria).

Fecal disgroup analysis is a study of fecal matter for the presence of pathogens of intestinal infections. This group includes bacteria that are not permanent inhabitants of the intestines of healthy children and can potentially cause an infectious process. These include the causative agents of dysentery (Shigella) and salmonella.

Opportunistic microorganisms (enterococci, staphylococci, clostridia, fungi) along with “beneficial” bacteria make up the natural intestinal microflora. They are called conditional because their ability to cause infection depends on specific circumstances: weakened immunity, a sharp decrease in the number of “helper bacteria” (lactobacteria, bifidobacteria), spoiled food.

In what cases is it necessary to examine stool?

It is necessary to submit stool for a disgroup test to clarify the diagnosis if an intestinal infection is suspected or as part of preventive measures.

For an intestinal infection to develop, pathogenic microbes must enter the child’s intestines. The source of infection is the feces of an infected person. Transmission of germs can occur through contact with a sick person or carrier of infection, or through consumption of contaminated food or water. The waterway is considered the most common.

Not all bacteria manage to reach the intestines - gastric juice causes the death of most of them. Since infants (especially newborns) produce small amounts of gastric juice, their risk of infection is higher. An infant may develop an intestinal infection when complementary foods are added. If complementary foods are prepared without following sanitary rules or have not undergone heat treatment, then opportunistic microbes may be present in it.

With an intestinal infection, a child may experience the following complaints:

• stomach ache;
• nausea, vomiting;
• loose stools;
• increased body temperature, chills, general weakness.

Acute intestinal infection in children can occur in the form of gastritis, enteritis or colitis, and it develops quickly. Diarrhea is a protective reaction of the body to remove the pathogen. But along with loose and copious stools, useful substances are also lost and, if treatment is not started in time, the child develops dehydration from excessive diarrhea and vomiting. The skin and visible mucous membranes become dry, the child becomes lethargic, the amount of urine decreases, and there is no sweat or tears. A newborn's fontanelle sinks. In very severe cases, shock and multiple organ failure develop. In infants, dehydration develops much faster.

The indication for analysis is to identify carriage. These are cases when children who have already had an intestinal infection continue to excrete a pathogenic microbe in their feces. Therefore, stool analysis for the intestinal group for hospitalization is an indispensable procedure if the child needs to undergo surgery and hospital treatment for another disease.

Preventive testing for carriage of intestinal pathogens is also carried out among medical practitioners, food industry workers and educational institutions and is included in the mandatory annual examination.

Can show whether necessary digestive enzymes are present in the body.

How to get tested? Rules for stool collection

Before you donate stool to the intestinal group, you need to know how to do it correctly. The technique of taking is not complicated. Feces should be collected in a special plastic bottle, which can be purchased at a pharmacy. Before collecting stool, you need to prepare the child: do not take activated charcoal, castor oil for several days, do not use rectal suppositories, and do not take antibiotics. They take feces after the child goes to the toilet “little by little.”

Collect feces from several places and fill one third of the container, this is about 2 cm. If you have to wait a long time for natural bowel movements, then feces taken from underwear (necessarily fresh) will be suitable as material for analysis. You can use a dropper to collect stool if the stool is very loose. Excrement is collected from areas where there are a lot of pathological impurities, such as pus, mucus, flakes. The selected material should not contain blood. We collect feces in the morning. There are cases when stool collection is not required; a rectal smear is sent for analysis. It is taken in the clinic with a special swab with the child lying on his side. The collected stool or smear is sent to the laboratory within 3 hours after collection.

Laboratory diagnostics

Analysis of intestinal infection in children includes bacterioscopic and bacteriological examination of stool. The bacterioscopic method shows the presence of bacteria and protozoan microorganisms in stool under a microscope. Bacteriological analysis of stool is the cultivation of a colony of microorganisms.

To do this, a tank is used to inoculate stool on a nutrient medium. The type of infectious pathogens and their concentration are determined by the nature of growth. When pathogenic microorganisms are detected, their sensitivity to antibiotics is determined.

The most frequently asked question: “How long does it take to test for intestinal infections?” It takes time for a colony to grow. How many days the culture will grow depends on the specific type of microbe. Data are usually received within 1–2 weeks.

Sometimes an analysis of the intestinal group may not be informative enough. For example, if the cause of the infection is a virus or protozoan microorganisms, or if not the pathogen itself, but its waste was found in the stool. In this case, feces are tested for polymerase chain reaction. Stool PCR detects the DNA of the pathogen, even if only one bacteria has entered the stool. Data arrives within a day.

Additional diagnostic methods for intestinal infections include a blood test, blood culture (if the child’s fever lasts more than 3 days) and serological analysis (allows us to identify antibodies to pathogens).

Stool culture for pathogenic intestinal flora also helps differentiate intestinal infection from other conditions that may cause the same symptoms. Differential diagnosis of intestinal infection is carried out with dysbiosis and food toxic infections. To diagnose dysbiosis, a stool test is performed for UPF.

Decoding the received data

A healthy child does not have pathogenic microorganisms in his stool, i.e. the analysis must be negative for disgroup and salmonellosis, as well as for dysenteric amoeba and typhoparatyphoid group. Normally, such simple microorganisms as intestinal trichomonas and balantidium are absent.

High levels of pathogenic bacteria that are not part of the normal microflora are found in two cases: acute intestinal infection and bacterial carriage.

A UPF test will tell you how many of these bacteria are in 1 gram of feces. Low values ​​of enterococci, clostridia, candida and bacteroides are considered a negative result, because For the development of infection, a certain amount of the pathogen is necessary, and not just its presence. If opportunistic microflora prevails over the concentration of lactobacilli in the feces, this is dysbacteriosis.

If an intestinal group is detected, the analysis is supplemented with a test for the sensitivity of bacteria to antibiotics.

Antibiotic sensitivity is indicated by the following letter designations

• S - sensitive (h);
• R - stable, resistant (y);
• I - moderately stable (yy).

Treatment

If the stool test comes back positive, then the sick child needs to be isolated. For mild cases of the disease, treatment can be carried out at home. It is necessary to restore the amount of fluid lost in feces (rehydration). The doctor prescribes antibiotic therapy. In case of severe dehydration, the child should be hospitalized immediately!

Differential diagnosis can be difficult if one disease occurs under the guise of another. In such cases, an accurate diagnosis is established based on the results of successful treatment. If the therapy is effective, the diagnosis is correct.

It is quite simple, however, at the same time, very informative analysis - it allows you to diagnose almost all diseases of the digestive system, clarify the location of the lesion (stomach, intestine or colon) and detect the presence of uninvited guests - parasites and pathogenic microbes. However, most parents by stool analysis only mean “test for dysbacteriosis,” which they love to prescribe to all children. However, there are several stool tests and microbiological culture (“for dysbacteriosis”) is performed as the last thing, when all other problems are excluded.

In children of the first year of life, stool analysis can identify various digestive problems - disturbances in the functioning of the stomach. Intestines, problems with the liver and pancreas, infectious diseases and hereditary metabolic pathologies. What tests are taken and what do they look at?

This analysis evaluates the main characteristics of feces - physical, chemical and microscopic; in addition, depending on the type of feeding and the age of the baby, it will have characteristic features. But in order for the analysis to show reliable data, it is necessary to collect it correctly, which is a rather difficult task for young children.
The analysis is collected from a diaper or oilcloth, since the diaper absorbs some of the liquid and the analysis will be unreliable.

For analysis, you need at least 1-2 teaspoons of feces, collected in a dry, clean container. But you should not use baby food jars - microscopic food particles remain on their bottom and walls, which will distort the result. Ideally, this should be a fresh analysis - the baby has had a full day, you have collected and delivered the analysis to the laboratory. But if this is not possible, collect it in a container, close the lid tightly and place it in the refrigerator door - you can store it for a maximum of 6-8 hours.
It is unacceptable to collect material after an enema or the use of laxative products or medications; there should be no urine particles in the stool.

How is it assessed and what should be normal?
Consistency. For children of the first two years of life, stool is acceptable in the form of porridge and unformed. On full breastfeeding, liquid porridge, with a small amount of water, maybe thick sour cream. On IV, the stool may be a soft sausage or mush. With the introduction of complementary feeding, the stool begins to take shape and becomes thicker; by the age of two it is usually a loose sausage. Stool in the form of “dry smoked sausage” or “sheep balls” is constipation. The stool is loose, watery with impurities, and often results in constipation.

Color. In babies of the first year of life, the stool is usually yellow, with small acceptable admixtures of green in the first 3-4 months. The stool then becomes yellow-brown, and by the age of two years it becomes approximately the same as that of adults. Pieces of food are acceptable, especially in the first months of complementary feeding, and when consuming brightly colored foods, stool should be colored in the color of the product (beets, rhubarb, pumpkin).
Smell. In the first months of life, the stool of breastfed babies has the smell of sour milk, while that of bottle-fed babies has a stronger smell. With the introduction of complementary foods, especially meat, the stool acquires the usual fecal smell.

RN (reaction). A neutral or slightly alkaline reaction is normal, but in the first months, when feeding with formula or breast milk, it can also be slightly acidic.
Protein. healthy children should not have protein in their stool - its presence indicates intestinal inflammation, the presence of mucus, bleeding, exudate, and undigested food.
Reaction to occult blood. Normally, there should be no blood in a healthy intestine - this always indicates problems. Most often, blood appears due to allergies and intestinal inflammation. rectal fissures. rectal prolapse, hemorrhoids, polyps and intestinal malformations.
Reaction to bilirubin. Bilirubin is one of the breakdown products of hemoglobin, which is present in feces until about three months of age; later, under the influence of enzymes and microflora, it is completely transformed into stercobilin, which gives a brown color. Its appearance in feces later than this age indicates health problems.

Slime. Mucus is a jelly-like discharge that is clear or whitish in color. They are designed to protect against the aggressive effects of intestinal contents. However, normally little mucus is secreted, and at the exit it is completely mixed with feces. A noticeable presence of mucus in feces is acceptable up to 3-5 months of life, especially in infants. And after that they talk about the presence of an inflammatory process.
Leukocytes. A small number of leukocytes, especially in the first months of life, is acceptable, however, if they are released in large numbers in combination with mucus and blood, this indicates inflammation and damage to the intestinal wall.

Muscle fibers. They talk about the degree of digestion of protein foods; before the introduction of meat complementary foods, the child practically does not have them, and in the future they should be sporadic. If there are a lot of them, this indicates digestive disorders in the stomach and intestines, problems with the pancreas, and inflammation of the intestines.

Connective tissue. Normally, it should not exist; it occurs only after the introduction of complementary foods with insufficient gastric secretion or problems with the pancreas.
Neutral fat. Fatty acid. Soap. In early childhood, small amounts of it can occur, but after six months it appears when the pancreas, liver, or diseases of the small intestine are disrupted.

Remains of undigested food. In the first years of life, the presence of plant food residues in feces is acceptable - especially corn, carrots, apple skins, etc. If there is a lot of digestible fiber in the stool, this may indicate that food is passing through the intestines too quickly. Undigested fiber may be normal.

Starch. It appears in food with the introduction of complementary foods; if there is a lot of it, this indicates poor functioning of the pancreas, indigestion in the intestines, and intestinal infection.
Iodophilic flora. These are special types of bacteria, the excessive growth of which occurs with dyspepsia (digestion disorders), disruption of the small intestine, disruption of the stomach and pancreas.

Fecal carbohydrate test.
This study is carried out if lactase deficiency is suspected - a decrease in the amount of the enzyme that breaks down milk sugar (lactose) in the intestines. It is usually prescribed for dyspeptic symptoms - loose, foamy stools, weight gain problems that occur when consuming dairy products. True lactase deficiency (when there is a deficiency of enzymes) is rare. Most often, there are false or secondary variants of lactase deficiency, when the enzyme is present, but it is either not yet mature, or is blocked as a result of an intestinal infection, or an overload of milk sugar occurs (as, for example, with an imbalance of foremilk and hindmilk in infants).

This condition goes away after correcting nutrition, treating intestinal infections and normalizing the microflora. Analysis of stool for carbohydrates is not the main thing in making a diagnosis, but it helps to determine the nature of the deficiency - primary or secondary.
To properly collect stool for determination of carbohydrates in stool, it is necessary to collect it in a clean, dry container, in a volume of at least 1-2 teaspoons. Feces must be delivered to the laboratory no later than 3-4 hours, otherwise the result will be distorted. The following are considered normal values ​​for children under one year of age:
On breastfeeding (before complementary feeding) up to 0.5-0.6%,
On artificial feeding - up to 0.3%,
For children from the second half of the year – up to 0.25%,
Over a year old 0%.

If indicators increase, feeding correction is prescribed, milk imbalance is eliminated if this is a baby, or a low-lactose formula is temporarily given if this is a baby on IV. If primary lactase deficiency is confirmed, treatment will consist of the use of the enzyme lactase.

Fecal analysis for opportunistic pathogenic flora (OPF).
It is this stool analysis, usually called “for dysbacteriosis,” that raises the greatest number of questions among parents. However, for the most part this analysis is uninformative and not indicative. The thing is that the intestinal microflora is very fickle and even if two tests are performed in a row on the same child in the morning and evening, they will be significantly different.

To understand where microbes come from in the intestines, and who should be feared and who not, let’s briefly talk about how the intestines work. Immediately after birth, when the baby is put to the breast, the baby’s intestines are populated with microflora from the mother. It becomes like a multi-storey building - there are peaceful neighbors and the majority of them are bifido- and lactoflora. And there are violent neighbors, but they are a minority, this is the so-called opportunistic flora (staphylococcus, Klebsiella, certain types of E. coli). While all the microbes are settling in the intestines, anxiety occurs - physiological dysbiosis, until everyone takes their rightful places. In addition, the settlement and peaceful existence in the intestines of various potentially dangerous microbes, such as staphylococcus and Klebsiella, is facilitated by help from the mother - breast milk with antibodies to harmful microbes and special substances that help the growth of beneficial lactobacilli and bifid flora. That is, all changes in the analysis of stool for microflora in the first 4-6 months of life are a physiological adjustment to the work and peaceful coexistence of microbes.

What is acceptable and what is not?
This analysis contains some special instructions, so the presence of the causative agent of salmonellosis, cholera, typhoid fever, dysentery and a pathogenic type of E. coli is definitely unacceptable in stool. But parents, however, are more concerned about the presence of staphylococcus, Proteus, Klebsiella, and enterococci in the stool. However, the mere presence of any amount of these microbes, in the absence of a clinical picture of these diseases, that is, if the child does not have a fever, gains weight, grows and develops according to age, is a variant of the norm for this baby. Microflora is called opportunistic because it can become active and harmful only under certain conditions.

This occurs in children with immunodeficiencies, a sharp decrease in immune strength after severe and long-term illnesses, after irradiation or treatment with heavy drugs - long-term antibiotics (three or more weeks), antitumor drugs. Another reason for the activation of this flora may be intestinal infections and severe poisoning - they simply join the clinical picture of the disease. In all other cases, correction of the microflora is not required at all - the microbial balance will be restored on its own. The use of any drugs to correct the flora is not required - most of them simply do not reach the intestines, being destroyed by hydrochloric acid of the stomach or intestinal enzymes.

Of course, this is not all of the stool tests that can be performed on a child. However, all other studies are quite specific and are carried out only if significant deviations are detected in the general analyzes. They are prescribed by a gastroenterologist and are necessary to clarify the diagnosis or to monitor the effectiveness of treatment.

It is known that microorganisms, despite their “small growth,” also have food “predilections,” an optimum temperature, in general, an environment that is ideal for them, where they feel comfortable and good, and therefore begin to multiply and grow intensively.

Bacteriological seeding or, as it is commonly called in short, tank seeding, is used to obtain a large number of microbes of one type (pure culture) in order to study their physicochemical and biological properties, so that later use the obtained data to diagnose infectious diseases.

Unfortunately, even the currently popular and other methods, the main disadvantage of which are false positive or false negative results, cannot always identify the pathogen. In addition, they are not able to select targeted antibacterial drugs. A similar problem is solved by a sowing tank, which is often in no hurry to prescribe, citing the fact that, for example, it is slowly cultivated, and the cost of analysis is considerable. However, health is worth it!

Conditions are needed for nutrition and breathing

Microbiologists now know that each pathogen needs its own “native” environment, taking into account its pH, redox potentials, viscosity, humidity and osmotic properties. Media can be soft and hard, simple and complex, universal and not very universal, but in all cases they must provide nutrition, respiration, reproduction and growth of the bacterial cell.

an example of the growth of microorganisms after tank sowing in a nutrient medium

Some media (thioglycolate, Sabouraud) are suitable for a wide range of microorganisms and are called universal. Others are intended only for certain species, for example, pneumococcus and Staphylococcus aureus, which produce hemolysins, grow on blood agar, which serves to isolate particularly “capricious” and, at the same time, dangerous strains. Thus, there are many types of media, where each of them grows its own range of microorganisms.

The purpose of cultivating microorganisms and its significance for diagnosis

In addition to water, air, soil, which contain various microorganisms in varying concentrations, including those that bring disease (pathogenic), many branches of medical science are interested in microbes living on the skin and mucous membranes of the human body, which can be represented by:

• Permanent inhabitants who do not pose any danger to humans, that is, the normal microflora of the body, without which we simply cannot live. For example, the disappearance of bacteria living in the intestines and participating in the digestion process leads to dysbiosis, which is not easy to treat. The same thing happens with the disappearance of vaginal microflora. It is immediately populated by opportunistic microorganisms, gardnerella, for example, which cause;
• Opportunistic pathogenic flora, which is harmful only in large quantities under certain conditions (immunodeficiency). The above-mentioned gardnerella is a representative of this type of microorganism;
• Presence of pathogenic microbes, which are not present in a healthy body. They are alien to the human body, where they enter accidentally through contact with another (sick) person and cause the development of an infectious process, sometimes quite severe or even fatal. For example, a meeting with pathogens - no matter what, it is treated at first, but (God forbid!) it will release cholera, plague, smallpox, etc.

Fortunately, many of them have been defeated and are currently kept under seal in special laboratories, but humanity must be prepared at any moment for the invasion of an invisible enemy capable of destroying entire nations. Bacteriological culture in such cases plays, perhaps, the main role in identifying the microorganism, that is, determining the genus, species, type, etc. (toxiconomic position), which is very important for the diagnosis of infectious processes, including sexually transmitted diseases.

Thus, sowing methods, like nutrient media, are different, however, they have the same goal: obtain a pure culture without foreign impurities in the form of microbes of other classes, which live everywhere: in water, in the air, on surfaces, on and inside humans.

When is tank sowing prescribed and how to understand the answers?

Name of microorganism and its quantity

Patients do not prescribe bacteriological analysis to themselves; this is done by the doctor if he has suspicions that the problems of a patient presenting various complaints are associated with the penetration of a pathogenic pathogen into the body or with the increased reproduction of microorganisms that constantly live with a person, but exhibit pathogenic properties only in certain conditions. Having passed the test and after some time received an answer, a person gets lost and sometimes gets scared when he sees incomprehensible words and symbols, therefore, to prevent this from happening, I would like to give a brief explanation on this issue:

When examining biological material for the presence of pathogenic microorganisms, the answer can be negative or positive (“bad tank culture”), since the human body is only a temporary shelter for them, and not a natural habitat.

Sometimes, depending on what material is to be inoculated, you can see the number of microorganisms expressed in colony-forming units per ml (one living cell will grow a whole colony) - CFU/ml. For example, culture of urine for bacteriological examination under normal conditions gives up to 10 3 CFU/ml of all identified bacterial cells, in doubtful cases (repeat the analysis!) - 10 3 - 10 4 CFU/ml, in case of an inflammatory process of infectious origin - 10 5 or higher CFU /ml. About the last two options in colloquial speech, sometimes they are simply expressed: “Bad tank sowing.”

How to “find control” against a pathogenic microorganism?

Simultaneously with the inoculation of the material in such situations, the microflora is inoculated for sensitivity to antibiotics, which will give a clear answer to the doctor - which antibacterial drugs and in what doses will “scare” the “uninvited guest”. There is also a decryption here, for example:

• The type of microorganism, for example, is the same E. coli in an amount of 1x10^6;
• The name of the antibiotic with the designation (S) indicates the sensitivity of the pathogen to this drug;
• The type of antibiotics that do not act on the microorganism is indicated by the symbol (R).

Bacteriological analysis is of particular value in determining sensitivity to antibiotics, since the main problem in the fight against chlamydia, mycoplasma, ureaplasma, etc. remains the selection of effective treatment that does not harm the body and does not impact the patient’s pocket.

Table: Alternative example of tank culture results identifying effective antibiotics

Proper preparation for bacteriological analysis is the key to reliable results

Any biological material taken from a person can be subjected to bacteriological analysis(skin, blood, sperm, mucous membranes of the oral cavity, respiratory and genitourinary tracts, gastrointestinal tract, organs of vision, hearing and smell, etc.). Most often, tank culture is prescribed by gynecologists and urologists, so we should dwell on it a little.

Proper preparation for bacteriological culture will be the key to the correct result, because otherwise, the analysis will have to be taken again and wait for the appointed time. How to donate blood from a vein for sterility is the task of health workers. As a rule, nothing depends on the patient here; he simply provides the elbow bend, and the nurse takes the sample into a sterile tube in compliance with all the rules of asepsis and antisepsis.

Another thing is urine or from the genital tract. Here the patient must ensure the first stage (collection), following the prescribed rules. It should be noted that the urine of women and men is somewhat different, although in the bladder of both sexes it is sterile:

• In women, when passing through the urethra it can capture a small number of non-pathogenic cocci, although in general it often remains sterile;
• For men, things are a little different. The anterior part of the urethra can supply passing urine with:
  1. diphtheroids;
  2. staphylococci;
  3. some non-pathogenic gram-negative bacteria, as will subsequently be shown by bacteriological analysis.

However, if they are in an acceptable concentration (up to 10 3 CFU/ml), then there is nothing to be afraid of, this is a variant of the norm.

To avoid the presence of other microorganisms and to ensure maximum sterility of the material taken, before the analysis, the genital organs are thoroughly cleaned (the entrance to the vagina in women is closed with a cotton swab - protection from the ingress of genital secretions). For analysis, an average portion of urine is taken (the beginning of urination into the toilet, approximately 10 ml average portion in a sterile jar, ending in the toilet). Patients need to know: urine taken for culture must be processed no later than two hours when stored at no higher than 20°C, so transportation time should be calculated.

In addition, material for the culture tank, if necessary, is taken from the urethra and rectum in men, from the urethra, rectum, vagina, cervix and cervical canal in women, but this happens in the medical institution where the patient must arrive. Washing, douching and using antiseptics in such cases is prohibited.

Other issues of concern to patients

Many patients are interested in how many days the analysis is done. This question cannot be answered unequivocally; it all depends on what material is being studied and what pathogen should be looked for. Sometimes the answer is ready in 3 days, sometimes in a week or even 10 - 14 days, since some samples require subculture to another medium.

People heading to tank sowing do not ignore the question of the price of analysis. The approximate cost in Moscow is about 800 – 1500 rubles. Of course, it can be higher and depends on the breadth of the spectrum of bacteriological search. You can probably get a free test during pregnancy at a antenatal clinic, or at a clinic for special medical reasons.

For pregnant women, tank sowing is mandatory, it is given 2 times(at registration and at 36 weeks), in this case, a smear is taken not only from the genital tract, but also from the mucous membranes of the nose and pharynx. The object of search in this case, in addition to urogenital infections, will be Staphylococcus aureus (Staphylococcus aureus), which in the postpartum period can cause a lot of trouble (purulent mastitis, etc.). In addition, pregnant women are required to undergo urine culture, scraping of the vaginal epithelium and smears from the cervix and cervical canal.

Many women, before going for the procedure, are very afraid of such terrible words and begin to think: “Is this necessary? Maybe I won’t go.” We hasten to assure you that the tests are absolutely painless. A smear from the cervix and cervical canal is taken with a sterile cytobrush, without causing the woman absolutely any pain, but subsequently a tank of inoculation from the w/m and c/c will protect both the expectant mother and the fetus from possible complications. The objects of search during pregnancy are the causative agents of chlamydia, urea and mycoplasma, yeast-like (usually Candida albicans), and other opportunistic and pathogenic microorganisms.

Video: video demonstration on tank culture from the cervical canal

Special cases of particular interest to those taking tests

Once pathogenic microorganisms enter the genital tract, they take hold within a very short time and begin their harmful activities. For example, always pathogenic gonococci (Neisseria), which are the culprits of a rather unpleasant disease called and related to STDs, feel “at home” literally on the 3rd day. They begin to actively reproduce and boldly move upward along the reproductive tract, capturing more and more new territories. Everyone knows that gonorrhea can now be treated well and almost no one is afraid of it anymore. But first you need to find her. The main method of searching for this infection is culture, culture, identification using Gram staining, and microscopy.

“Coffee beans” (diplococci) found in pairs in a smear taken “for flora” from the genital tract do not indicate the presence of a sexually transmitted disease. Such vaginal microflora often appears in postmenopause and does not mean anything bad. A smear taken under non-sterile conditions on a glass slide and stained with methylene blue or Romanovsky (cytology) cannot differentiate the microorganism. He can only make a guess and refer the patient for additional research (obtaining an isolated culture).

It should be noted that while scrapings from the mucous membranes of the genitourinary tract taken for culture for ureaplasma are not such a rare occurrence, then doctors themselves often avoid urine culture, since it is more difficult to work with.

It creates difficulties in diagnosis, causing great harm not only during pregnancy. In addition, chlamydia causes many diseases that are characteristic not only of women, but also of the male population, so it is sown, cultivated, studied, sensitivity to antibacterial therapy is determined and, thus, it is combated.

During pregnancy, it is generally difficult to do without bacteriological culture, since many microorganisms, masked in a cytological smear, can be missed. Meanwhile, the effect of some STD pathogens on the fetus can be detrimental. In addition, treating a pregnant woman is much more difficult, and prescribing antibiotics “by eye” is simply unacceptable.

Sowing methods

To isolate pure cultures of pathogens, the first stage is to inoculate them on appropriate media, which is carried out under special (sterile!) conditions. Basically, the transfer of material to the medium is carried out using devices used back in the 19th century by the great Louis Pasteur:

• Bacterial loop;
• Pasteur pipette;
• Glass rod.

Of course, many instruments have undergone changes over 2 centuries, replaced by sterile and disposable plastic ones, however, the old ones have not remained in the past, continuing to serve microbiological science to this day.

The first stage of obtaining colonies requires compliance with certain rules:

1. Sowing is carried out over an alcohol lamp in a box pre-treated with disinfectants and quartz treatment, or in a laminar flow hood, ensuring sterility in the work area;
2. The health worker's clothing, gloves and environment must also be sterile, since the opposite interferes with the isolation of isolated strains;
3. You need to work quickly but carefully in the box; you cannot talk or be distracted; at the same time, you must remember about personal safety, because the material can be infectious.

Isolation of strains and study of pure cultures

The isolation of strains is not always the same, since some biological media found in the human body require an individual approach, for example, hemoculture (blood) is first “grown up” in a liquid medium (ratio 1: 10), since blood (undiluted) can kill microorganisms, and then, after a day or more, they are transferred to Petri dishes.

Sowing urine, gastric lavage waters and other liquid materials also has its own characteristics, where in order to obtain a pure culture, the liquid must first be centrifuged (aseptic conditions!), and only then sowed, not the liquid itself, but its sediment.

Cultivation and growing of colonies is carried out on Petri dishes or placed first in a liquid medium poured into sterile bottles, and then the isolated colonies are sown again, but on slanted agar and the material is placed in a thermostat for a day. After making sure that the resulting culture is pure, the strains are transferred to a glass slide, a smear is made and stained with Gram (most often), Ziehl-Neelsen, etc., and for differentiation, the morphology of the microbe is studied under a microscope:

• Size and shape of the bacterial cell;
• Presence of capsules, flagella, spores;
• Tinctorial properties (relationship of microorganism to staining)*.

*The reader has probably heard of such a pathogen as treponema pallidum? This is the causative agent of syphilis, and its name (pale) is why it appears that it does not perceive paint well and remains slightly pinkish when stained according to Romanovsky. Microorganisms that do not accept aniline dyes are called gram-negative, and those that perceive are called gram-positive. Gram-negative bacteria are given a pink or red color when stained with Gram by additional dyes (fuchsin, safranin).

Tank culture can be called an ancient analysis, but its popularity does not decrease because of this, although modern bacteriology has the ability to isolate not only strains, but also a separate cell from it, which is called clone. However, to obtain a clone, a special device is required - a micromanipulator, which is not available in ordinary laboratories, since it is used mainly for research purposes (genetic research).

> Culture of pathogenic intestinal flora, determination of its sensitivity to antimicrobial drugs and bacteriophages

This information cannot be used for self-medication!
Consultation with a specialist is required!

The human intestine is normally populated by a large number of microorganisms. These are various representatives of lactobacilli, bacteroides, enterobacteria, peptostreptococci, etc. These bacteria make up the so-called intestinal microflora, participate in the digestion process, support local immunity, preventing the development of infections. Intestinal infections develop when pathogenic (disease-causing) microorganisms enter the intestinal lumen and multiply. The most common pathogens of intestinal infections are Shigella and Salmonella. Vibrio cholerae, clostridia (causing botulism, etc.), Yersinia, and Staphylococcus aureus are detected somewhat less frequently. Culture for pathogenic intestinal flora determines the cause of intestinal infection, and determining the sensitivity of microorganisms to antibiotics and bacteriophages helps to choose the right therapy.

What are the manifestations of acute intestinal infections?

Symptoms and complaints vary somewhat depending on the causative agent of the infection. When infected with Shigella, the onset of the disease is acute. Cramping pain in the abdomen occurs and the temperature rises. Characterized by frequent, painful urges to defecate, not accompanied by the passage of feces, or feces are separated in small quantities mixed with mucus and blood. Salmonellosis is characterized by the simultaneous occurrence of vomiting and diarrhea. The stool may be yellowish or greenish. Pain and rumbling in the abdomen are more pronounced to the right of the navel. All intestinal infections accompanied by diarrhea or vomiting lead to intoxication and dehydration. This is manifested by dry skin and mucous membranes, and in severe cases – loss of body weight.

When is culture for pathogenic intestinal flora prescribed?

The study is prescribed by an infectious disease specialist, therapist or pediatrician if an acute intestinal infection is suspected. In this case, culture with determination of sensitivity to antibiotics and bacteriophages helps to identify the causative microorganism, carry out differential diagnosis with other diseases that have similar symptoms, and select the correct therapy. At the end of treatment, re-sampling of material for culture is required in order to control the destruction of the pathogen.

The analysis is carried out when preparing personal medical records for workers in public spheres (food, healthcare, education, etc.). If bacterial carriage is detected, therapy is carried out aimed at destroying the microorganism, after which the analysis should be retaken.

How is the study carried out, and how to properly prepare for it?

The study is carried out without preliminary preparation. The material for bacteriological culture is fresh feces. It is lifted into a tightly closed sterile container with a spatula. Defecation before this is carried out in a clean vessel or pot. When collecting material, you should not contaminate it with urine and secretions from the genital tract. If there is mucus and pus in the stool, it is advisable to select such fragments of it, since the concentration of the pathogen in them is much higher.

Material can be collected from the rectum with a sterile swab. After inserting the tampon into the anus, perform rotational movements in both directions, and the resulting material is transferred to a sterile container.

How are the results interpreted?

Normally, there should be no growth of pathogenic microorganisms during sowing. If growth is detected, we can talk about an acute intestinal infection or bacterial carriage. Determining sensitivity to antibiotics and bacteriophages in this case will help you choose the right drug for treatment.

If there are few pathogenic bacteria in the stool, the result may be negative. An indirect sign of the presence of pathogens may be a decrease in the growth of normal flora.

The human intestine is inhabited by ~3 kg of bacteria. They represent the microflora necessary for normal digestion, but in the event of various malfunctions, the number of some microorganisms can noticeably decrease - dysbacteriosis will occur - an imbalance of bacteria.

Although doctors do not classify it as an independent disease, this does not reduce the harm. If intestinal dysbiosis is suspected, special stool tests are prescribed to establish an accurate diagnosis.

Microorganisms living in the gastrointestinal tract are of paramount importance for human life. They synthesize vitamins, break down food and protect against attacks by pathogenic strains.

In other words, humans and bacteria are in symbiosis. But if the composition of the microflora is disturbed, flatulence, diarrhea, nausea may occur, not to mention the consequences of insufficient supply of nutritional compounds to the tissues.

The purpose of stool analysis is to determine the qualitative and quantitative composition of bacteria in the intestines.

For this purpose, 3 methods are usually used in medicine:

1. Coprogram.
2. Bacteriological analysis.
3. Biochemical analysis.

Coprogram

A coprogram is prescribed when a person complains of chronic or acute stool disorder, abdominal pain of unknown origin, or sudden weight loss for no apparent reason.

Doctors also resort to such research when treating diseases not related to the gastrointestinal tract. This is especially true when treating pathologies in different parts of the body with antibiotics (throat, joints, etc.).

A coprogram is a primary examination, which is only an auxiliary method and gives a physical characteristic of the intestinal contents.

The analysis is carried out in 2 stages:

2. Microscopic:

• cells and tissue fragments;
• digested food (fiber, fat, salts, starch, etc.).

If the coprogram shows deviations from the norm, the doctor has a reason to conduct a more thorough analysis. In the laboratory, stool is cultured on a nutrient medium.

After 4-5 days, the bacteria will multiply, which will allow their colonies to be examined under a microscope. After this, the specialist makes a conclusion about the number of microbes in 1 g of feces (CFU/g).

Based on the data obtained, the doctor makes a diagnosis. Test results for adults and children often differ, so the patient’s age must be taken into account.

But waiting 5 days for colonies to grow is not always permissible, because during this time a person’s condition can deteriorate significantly.

Biochemical analysis of stool

Biochemical analysis of stool for dysbacteriosis gives results on the day the samples are submitted. The essence of such research is to identify compounds present in the intestines.

Particular attention is paid to the spectrum of fatty acids, because they are synthesized by bacteria in the process of life. Biochemical analysis is also called rapid diagnostics.

The method is very informative and simple; it not only demonstrates the imbalance of microflora, but also establishes the section of the intestine in which the malfunction occurred.

Doctors much more often prefer this study due to its significant advantages:

• Speed. Results will be available in 1-2 hours.
• Sensitivity. The method very accurately determines the concentration of compounds.
• Undemanding to the freshness of samples. Even yesterday's poop will do.

The reliability of the research results directly depends on proper preparation. The fact is that many food products contain substances that will give a positive reaction.

First of all, it's meat. It is in it that hemoglobin is present.

Secondly, it's iron. All red products contain this element. It is worth refraining from eating such foods for 3 days before the test, so that the laboratory does not accidentally get a false positive result.

Restrictions also apply to raw vegetables and fruits: during the preparation period you need to eat only thermally processed plant products.

In addition, the patient must stop taking medications that directly affect the intestinal microflora:

• antibiotics;
• probiotics;
• laxatives (official and popular);
• rectal suppositories.

Adults prepare for a stool test for dysbiosis on their own. Examining the contents of a child’s intestines is no different, but parents will have to monitor the child’s compliance with all recommendations.

How to properly take a stool test for dysbacteriosis?

Diet and medication withdrawal are the primary conditions for the reliability of the analysis results. In addition, the patient will need to collect stool in accordance with the rules.

Handing over feces - 6 rules:

1. Before the control bowel movement, wash the perineum (the possibility of getting old samples is eliminated).
2. The use of any auxiliary means to speed up the process of defecation (enema, laxative) is prohibited.
3. Prepare a special container with a tight lid in advance (must be purchased at a pharmacy).
4. Do not allow liquid to get into the stool (urine, water, etc.).
5. Take 3 stool fragments (1 teaspoon each from different areas).
6. If blood or mucus is present, such samples must be taken.

Gut bacteria are primarily anaerobic. 1 hour after defecation, they will still maintain their population in their natural form, but gradually the microorganisms will begin to die.

In order to correctly test stool for dysbacteriosis, it is necessary to deliver excrement samples to the laboratory at least within 2 hours after bowel movement.

Urgency is not so important for biochemical research, which studies not bacterial colonies, but the result of their vital activity - fatty acids. These compounds almost do not disintegrate spontaneously, and therefore remain unchanged for quite a long time.

Doctors even allow you to freeze stool and bring it in the next day. In the case of newborn children, this option is sometimes the most preferable for parents.

The intestines are home to 100 trillion bacteria, which is 10 times the number of all cells in the body. If there are no microbes at all, then the person will simply die.

On the other hand, a shift in balance in any direction leads to diseases. The interpretation of stool analysis for dysbiosis is to determine the number and types of microbes.

Table of interpretation of results and norms of analysis

Detailed transcript:

1. Bifidobacteria:

• 95% of all bacteria living in the intestines;
• synthesize vitamins K and B;
• promote the absorption of vitamin D and calcium;
• strengthen the immune system.

2. Lactobacilli:

• maintain acidity;
• synthesize lactase and protective substances.

3. Escherichia:

• synthesize vitamins K and B;
• promote the absorption of sugars;
• produce colicins, proteins that kill microbes.

4. Bacteroides:

• break down fats;
• perform a protective function.

5. Streptococci:

• break down carbohydrates;
• perform a protective function;
• present in small quantities and not always.

6. Enterococci:

• break down carbohydrates.

7. Peptococci:

• participate in the synthesis of fatty acids;
• perform a protective function;
• are not always present.

8. Staphylococci:

• live in the large intestine;
• participate in nitrate metabolism;
• There are many pathogenic strains.

9. Clostridia:

• live in the large intestine;
• synthesize acids and alcohols;
• break down proteins.

10. Fungal:

• maintain an acidic environment;
• opportunistic.

A change in the number of certain microorganisms is possible when pathogenic strains enter the intestines.

This usually occurs due to poor personal hygiene (dirty hands, unwashed fruits and vegetables). Antibiotic treatment is the second common cause of dysbiosis.

To normalize the situation in the gastrointestinal tract, doctors additionally prescribe probiotics - special dietary supplements.

In addition, dysbiosis often indicates an immune failure. Leukocytes control the population of microbes, the number of which increases significantly when natural defenses decrease. And often it is not beneficial bacteria that multiply, but pathogenic ones.

Stool analysis in children

The results of stool analysis for dysbacteriosis in children are somewhat different than in adults. This is due, first of all, to the gradual colonization of the intestines by microorganisms.

After birth, the child feeds on mother's milk, which promotes the development of normal microflora. But in hospitals, infection with Staphylococcus aureus often occurs.

And if the mother does not have antibodies to this microorganism, then the baby will develop dysbacteriosis.

In addition, some beneficial strains appear only within 1 year, such as bacteroides. Sometimes fungi of the genus Candida develop excessively in the child’s intestines, which provoke the corresponding disease - candidiasis.

The most common cause of dysbiosis in children is an early transition to artificial feeding. After all, the baby needs mother's milk in the first year of life.

Conclusion

A stool test for dysbacteriosis is prescribed for any digestive disorders. In addition, doctors monitor the state of the patient’s microflora during antibiotic therapy.

Timely identification of dysbiosis and clarification of the nature of the disorder will make it possible to take the right steps and reduce the likelihood of complications.