Anaerobes are bacteria. Features of anaerobes, classification and methods of cultivation

Overview of popular bacteria manufacturers and are they afraid of chemistry

When choosing which bacteria to use to clean septic tanks, give preference to compounds from the following manufacturers:

Dr. Robik;
Vodohray;
Waste Treat;
Micropan;
Saneks
Bioforce Septic.

Preparations "Vodogray" have a powdery consistency, bacteria are activated 20 minutes after dilution in water. The line consists of compounds based on aerobic organisms, for the vital activity of which it is necessary to ensure the flow of air into the septic tank.

The effect of the activity of microorganisms becomes noticeable after 1-2 weeks from the moment of addition. Complete processing of the contents of the septic tank occurs in 2-3 months. Microbes "Vodogray" are resistant to the effects of household chemicals. The cost of the drug is 330 rubles. for 100 gr.

Compositions "Doctor Robik" are classified into types according to their functional purpose: K57 - for especially contaminated septic tanks, K87 - bacteria for a septic tank that are not afraid of chemistry, K107 - a universal preparation, for general conditions of use. Preparations "Doctor Robik" are distinguished by a high speed of activation - the smell of a cesspool disappears 4 days after addition. The cost is 150 rubles. for 75 grams of powder.

Waste Treat is a combined preparation containing two types of bacteria. Sold in the form of tablets, granules or liquid concentrate. When used, 1 tablet is dissolved in a bucket of water, after which the liquid is added to the septic tank. The concentrate is diluted in a ratio of 1 to 200. The cost of the drug is 450 rubles per 10 ml. concentrate.

"Micropan" is a highly effective biological treatment agent. The recycled solid waste is subsequently used as fertilizer, water - for watering vegetable gardens. "Miropan" has the form of tablets (consumption of 1 piece per cubic meter of wastewater) and a liquid composition (500 ml of the drug is enough for a cesspool of a country toilet or a small septic tank for 3 months of constant cleaning). Price - 210 rubles. for 100 ml. In addition to products for septic tanks, the company's line includes a composition that accelerates the decay of compost pits.

"Sanex" is an American powdered drug sold in packages weighing 100 and 400 grams. The cost of 100 gr. packaging - 150 rubles, 400 gr. - 370 rubles. The company's line includes three compositions of different purposes - for septic tanks, compost pits and clearing blockages in sewer pipes.

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WATCH VIDEO

«Вioforce Septic» is a Canadian-made combined preparation, one of the most effective means for waste treatment. It is sold in packages weighing 670 gr., Consisting of 12 bags. At the loading stage, it is poured into the septic tank 1 sachet daily for 4 days, then - 1 sachet once a month for maintenance. The cost of packaging is 3 thousand rubles. Bacteria for septic tanks help to deal with waste without difficulty.

Classification

Anaerobic bacteria, in turn, are divided into 3 groups according to oxygen tolerance and need for it:

Optional - able to grow aerobically or anaerobically, i.e. in the presence or absence of O2.
Microaerophiles – require low oxygen concentration (eg 5%), and many of them require high CO 2 concentration (eg 10%); in the complete absence of oxygen, they grow very weakly.
Obligatory (mandatory, strict) –
are incapable of aerobic metabolism (grow in the presence of oxygen), but have different tolerance to O 2 (the ability to survive for some time).

Obligate anaerobes thrive in areas of low redox potential (eg, necrotic, dead tissue). Oxygen is toxic to them.There is a classification according to its portability:

Strict - withstand only ≤0.5% O 2 in air.
Moderate - 2-8% O 2.
Aerotolerant anaerobes - tolerate atmospheric O2 for a limited time.

The average percentage of oxygen in the earth's atmosphere is 21.

Differential diagnostic nutrient media

environments gissa
("variegated row")
Wednesday Ressel
(Russell)
Wednesday Ploskireva
or baktoagar "Zh"
Bismuth Sulfite Agar

Hiss media
: To 1% peptone water, add a 0.5% solution of a certain carbohydrate (glucose, lactose, maltose, mannitol, sucrose, etc.) and Andrede's acid-base indicator, pour into test tubes into which a float is placed to trap gaseous products formed during decomposition of hydrocarbons.

Ressel Wednesday
(Russell) is used to study the biochemical properties of enterobacteria (Shigella, Salmonella). Contains nutrient agar-agar, lactose, glucose and indicator (bromothymol blue). The color of the medium is grassy green. Usually prepared in 5 ml tubes with a beveled surface. Sowing is carried out by an injection into the depth of the column and a stroke along the beveled surface.

Wednesday Ploskirev
(bactoagar G) is a differential diagnostic and selective medium, since it inhibits the growth of many microorganisms and promotes the growth of pathogenic bacteria (causative agents of typhoid, paratyphoid, dysentery). Lactose-negative bacteria form colorless colonies on this medium, while lactose-positive bacteria form red colonies. The medium contains agar, lactose, brilliant green, bile salts, mineral salts, indicator (neutral red).

Bismuth Sulfite Agar
It is designed to isolate salmonella in its pure form from infected material. Contains tryptic digest, glucose, salmonella growth factors, brilliant green and agar. The differential properties of the medium are based on the ability of Salmonella to produce hydrogen sulfide, on their resistance to the presence of sulfide, brilliant green and bismuth citrate. Colonies are marked in black color of bismuth sulfide (the technique is similar to the medium Wilson-Blair
).

Diagnostics

Samples of anaerobic culture should be obtained by aspiration or biopsy from areas that do not normally contain them. Delivery to the laboratory must be prompt, and transport equipment must provide an anoxic environment with carbon dioxide, hydrogen, and nitrogen. Swabs are best transported in an anaerobically sterilized semi-solid medium such as Cary-Blair transport medium (a special solution that contains a minimum of nutrients to grow bacteria and substances that can kill them).

anaerobic organisms

The respiration and growth of aerobes is manifested as the formation of turbidity in liquid media or, in the case of dense media, as the formation of colonies. On average, it takes about 18 to 24 hours to grow aerobes under thermostatic conditions.

What it is

Anaerobes are always present in the normal microflora, mucous membranes of the body, in the gastrointestinal tract and genitourinary system. They are classified as conditionally pathogenic microorganisms, since they are natural inhabitants of the biotopes of a living organism.

With a decrease in immunity or the influence of negative factors, bacteria begin to actively multiply uncontrollably, and microorganisms turn into pathogens and become sources of infection. Their waste products are dangerous, toxic and rather aggressive substances. They are able to easily penetrate cells or other organs of the body and infect them.

In the body, some enzymes (for example, hyaluronidase or heparinase) increase the pathogenicity of anaerobes, as a result, the latter begin to destroy muscle and connective tissue fibers, which leads to microcirculation disorders. Vessels become fragile, erythrocytes are destroyed.All this provokes the development of immunopathological inflammation of blood vessels - arteries, veins, capillaries and microthrombosis.

The danger of the disease is associated with a large percentage of deaths, so it is extremely important to notice the onset of the infection in time and immediately begin its treatment.

What are anaerobic bacteria

Anaerobic bacteria are microorganisms that grow in the absence of oxygen. Bacteria that are unable to carry oxygen are called obligate anaerobes. Facultative anaerobes can grow without oxygen. But they are able to use oxygen, if it is available in the environment, to generate more energy than normal anaerobic respiration. Although aerotolerant bacteria do not use oxygen, they can survive in the presence of oxygen. Anaerobic bacteria play a major role in nutrition cycles such as the nitrogen cycle. Anaerobic bacteria in the nitrogen cycle and their role are shown in figure 2.

Figure 2: Nitrogen cycle

Some of the obligate anaerobes use fermentation while others use anaerobic respiration. Aerotolerant bacteria ferment strictly while facultative anaerobes use fermentation, anaerobic respiration, or aerobic respiration.

Fermentation

The two types of fermentation are lactic acid fermentation and ethanol fermentation. Both methods correspond to glycolysis. The second step is fermentation. The electronic transport chain is not used in the fermentation process. The chemical reactions for each type of fermentation are shown below.

ethanol fermentation

The final electron acceptor of anaerobic respiration is not molecular oxygen, as in aerobic respiration. Different types of organisms use different types of terminal electron acceptors. These can be ions such as sulfur, ferric iron, manganese (IV), cobalt (III) and uranium (VI), and compounds such as fumarate, sulfate, nitrate or carbon dioxide. Methanogenic bacteria are one such type of organism that use carbon dioxide as the final electron acceptor in the absence of oxygen. They produce methane gas as a by-product. Bacteroides, Clostridium, and E. coli are some examples of anaerobic bacteria.

Classification of anaerobes

According to the classification established in microbiology, there are:

Facultative anaerobes
Capneistic anaerobes and microaerophiles
Aerotolerant anaerobes
Moderately strict anaerobes
obligate anaerobes

If an organism is able to switch from one metabolic pathway to another (for example, from anaerobic respiration to aerobic respiration and vice versa), then it is conditionally referred to as facultative anaerobes
.

Until 1991, a class was distinguished in microbiology capneistic anaerobes
, requiring a reduced concentration of oxygen and an increased concentration of carbon dioxide (Brucella bovine type - B. abortus
)

A moderately strict anaerobic organism survives in an environment with molecular O 2 but does not reproduce. Microaerophiles are able to survive and multiply in an environment with a low partial pressure of O 2 .

If the organism is not able to "switch" from anaerobic to aerobic respiration, but does not die in the presence of molecular oxygen, then it belongs to the group aerotolerant anaerobes
. For example, lactic acid and many butyric bacteria

obligate
anaerobes in the presence of molecular oxygen O 2 die - for example, representatives of the genus bacteria and archaea: Bacteroides
, Fusobacterium
, Butyrivibrio
, Methanobacterium
). Such anaerobes constantly live in an oxygen-deprived environment. Obligate anaerobes include some bacteria, yeasts, flagellates and ciliates.

Types of anaerobic infection

Surgical infection or gas gangrene

increasing pain with a feeling of fullness, since the process of gas formation takes place in the wound;
fetid smell;
exit from the wound of a purulent heterogeneous mass with gas bubbles or inclusions of fat.

Anaerobic surgical infection is rare, and its occurrence is directly related to the violation of antiseptic and sanitary standards during surgical operations.

anaerobic clostridial infections

In this case, the pathogen enters the human body from the external environment. For example, these are such pathogens:

tetanus;
botulism;
gas gangrene;
toxicoinfections associated with the use of low-quality contaminated food.

In gynecology

The penetration of anaerobic infection into the female body is facilitated by:

injuries of the soft tissues of the vagina and perineum, for example, during childbirth, during abortions or instrumental studies;
various vaginitis, cervicitis, cervical erosion, genital tract tumors;
remnants of membranes, placenta, blood clots after childbirth in the uterus.

Causes of infection

There are several main reasons why infection occurs:

Creation of suitable conditions for the vital activity of pathogenic bacteria. This may happen:
when an active internal microflora gets on sterile tissues;
when using antibiotics that have no effect on anaerobic gram-negative bacteria;
in case of circulatory disorders, for example, in the case of surgery, tumors, injuries, ingestion of a foreign body, vascular diseases, and tissue necrosis.
Infection of tissue by aerobic bacteria. They, in turn, create the necessary conditions for the vital activity of anaerobic microorganisms.
Chronic diseases.
Some tumors that are localized in the intestines and head are often accompanied by this disease.

Toxicity of oxygen and its forms for anaerobic organisms

An oxygen rich environment is aggressive towards organic life forms. This is due to the formation of reactive oxygen species in the course of life or under the influence of various forms of ionizing radiation, which are much more toxic than molecular oxygen O₂. The factor that determines the viability of an organism in an oxygen environment is the presence of a functional antioxidant system capable of eliminating: superoxide anion (O₂−), hydrogen peroxide (H₂O₂), singlet oxygen (O.), as well as molecular oxygen (O₂) from the internal environment of the body.
Most often, such protection is provided by one or more enzymes:

superoxide dismutase, which eliminates the superoxide anion (O₂−) without energy benefits for the body
catalase that eliminates hydrogen peroxide (H₂O₂) without energy benefits for the body
cytochrome— enzyme responsible for the transfer of electrons from NAD•H to O₂. This process provides a significant energy benefit to the body.

Aerobic organisms most often contain three cytochromes, facultative anaerobes - one or two, obligate anaerobes do not contain cytochromes.

Anaerobic microorganisms can actively influence the environment
, creating a suitable redox potential of the medium (for example, Clostridium perfringens). Some inoculated cultures of anaerobic microorganisms lower the pH before they begin to multiply.₂ from value to , protecting themselves with a reductive barrier, others - aerotolerant - produce hydrogen peroxide in the process of life, increasing pH₂.

Additional antioxidant protection can be provided by the synthesis or accumulation of low molecular weight antioxidants: vitamin C, A, E, citric and other acids.

Aerophilic microorganisms

Aerobes are called microorganisms whose respiration is impossible without free oxygen in the air, and their cultivation takes place on the surface of nutrient media.

According to the degree of dependence on oxygen, all aerobes are divided into:

obligate (aerophiles) - able to develop only at a high concentration of oxygen in the air;
facultative aerobic microorganisms that develop even with a reduced amount of oxygen.

Properties and features of aerobes

Aerobic, water and air, and are actively involved in the cycle of substances. The respiration of bacteria, which are aerobes, is carried out by direct oxidation of methane (CH 4), hydrogen (H 2), nitrogen (N 2), hydrogen sulfide (H 2 S), iron (Fe).

Obligate aerobic microorganisms that are pathogenic to humans include tubercle bacillus, tularemia pathogens, and vibrio cholerae.
All of them require high levels of oxygen to survive. Facultative aerobic bacteria, such as salmonella, are capable of respiration with very little oxygen.

Aerobic microorganisms that carry out their respiration in an oxygen atmosphere are able to exist in a very wide range at a partial pressure of 0.1 to 20 atm.

Growing Aerobes

It implies the use of a suitable nutrient medium. The necessary conditions are also the quantitative control of the oxygen atmosphere and the creation of optimal temperatures.

Classification of anaerobes

According to the classification established in microbiology, there are:

Facultative anaerobes
Capneistic anaerobes and microaerophiles
Aerotolerant anaerobes
Moderately strict anaerobes
obligate anaerobes

Anaerobic training

Anaerobic exercises are performed without the participation of oxygen. They are characterized by high intensity and short duration. In this case, the maximum effort is made. These are a series of exercises that are broken down into short sets and performed at a fast pace.

Anaerobic exercises give the following results:

Increases strength and endurance.
The process of fat burning is accelerated due to the large number of calories expended.
Increases metabolism, strengthens and develops muscles.
With the observance of special diets, there is a set of muscle mass.
It is thanks to anaerobic exercises that you can create a beautiful muscular relief.
The musculoskeletal system is strengthened.
Increases immunity and improves well-being.

But, as with aerobic, you need to understand that all this only works in conjunction with proper nutrition and subject to regular workouts built according to the right program.

General culture methods for anaerobic organisms

Gaspack
- the system chemically ensures the constancy of the gas mixture acceptable for the growth of most anaerobic microorganisms.In a sealed container, water reacts with sodium borohydride and sodium bicarbonate tablets to form hydrogen and carbon dioxide. Hydrogen then reacts with oxygen in the gas mixture on a palladium catalyst to form water, which is already re-reacting with the hydrolysis of borohydride.

This method was proposed by Brewer and Olgaer in 1965. The developers introduced a disposable hydrogen generating sachet, which was later upgraded to carbon dioxide generating sachets containing an internal catalyst.

Zeissler method
used to isolate pure cultures of spore-forming anaerobes. To do this, inoculate on the Kitt-Tarozzi medium, heat it for 20 minutes at 80 ° C (to destroy the vegetative form), fill the medium with vaseline oil and incubate for 24 hours in a thermostat. Then, inoculation is carried out on sugar-blood agar to obtain pure cultures. After 24-hour cultivation, the colonies of interest are studied - they are subcultured on the Kitt-Tarozzi medium (with subsequent control of the purity of the isolated culture).

Fortner method

Fortner method
- inoculations are made on a Petri dish with a thickened layer of the medium, divided in half by a narrow groove cut in agar. One half is seeded with a culture of aerobic bacteria, the other half is inoculated with anaerobic bacteria. The edges of the cup are filled with paraffin and incubated in a thermostat. Initially, the growth of the aerobic microflora is observed, and then (after the absorption of oxygen), the growth of the aerobic microflora stops abruptly and the growth of the anaerobic microflora begins.

Weinberg method
used to obtain pure cultures of obligate anaerobes. Cultures grown on Kitta-Tarozzi medium are transferred to sugar broth. Then, with a disposable Pasteur pipette, the material is transferred into narrow tubes (Vignal tubes) with sugar meat-peptone agar, immersing the pipette to the bottom of the tube. The inoculated tubes are rapidly cooled, which makes it possible to fix the bacterial material in the thickness of the hardened agar. The tubes are incubated in a thermostat, and then the grown colonies are studied. When a colony of interest is found, a cut is made in its place, the material is quickly taken and inoculated on the Kitta-Tarozzi medium (with subsequent control of the purity of the isolated culture).

Peretz method

Peretz Method
- a culture of bacteria is introduced into the melted and cooled sugar agar-agar and poured under glass placed on cork sticks (or fragments of matches) in a Petri dish. The method is the least reliable of all, but it is quite simple to use.

Qualification of anaerobic infections according to the localization of its focus

There are the following types of anaerobic infections:

Soft tissue and skin infections
. The disease is caused by anaerobic Gram-negative bacteria. These are superficial diseases (cellulitis, infected skin ulcers, consequences after major diseases - eczema, scabies, and others), as well as subcutaneous infections or postoperative ones - subcutaneous abscesses, gas gangrene, bite wounds, burns, infected ulcers in diabetes, vascular diseases. With a deep infection, soft tissue necrosis occurs, in which there is an accumulation of gas, gray pus with a vile odor.
Bone infection
. Septic arthritis is often the result of neglected Vincent, osteomyelitis, a purulent-necrotic disease that develops in the bone or bone marrow and surrounding tissues.
Infections of the internal organs
, including women, bacterial vaginosis, septic abortion, abscesses in the genital apparatus, intrauterine and gynecological infections may occur.
Infections of the bloodstream
- sepsis. It spreads through the bloodstream;
Serous cavity infections
- peritonitis, that is, inflammation of the peritoneum.
bacteremia
- the presence of bacteria in the blood that get there by exogenous or endogenous means.

Anaerobes are bacteria.Features of anaerobes, classification and methods of cultivation

Toxicity of oxygen and its forms for anaerobic organisms

An oxygen rich environment is aggressive towards organic life forms. This is due to the formation of reactive oxygen species in the course of life or under the influence of various forms of ionizing radiation, which are much more toxic than molecular oxygen O 2 . The factor determining the viability of an organism in an oxygen environment is the presence of a functional antioxidant system capable of eliminating: superoxide anion (O 2 -), hydrogen peroxide (H 2 O 2), singlet oxygen (O .), and also molecular oxygen ( O 2) from the internal environment of the body. Most often, such protection is provided by one or more enzymes:

superoxide dismutaseeliminating superoxide anion (O 2 -) without energy benefits for the body
catalase, eliminating hydrogen peroxide (H 2 O 2) without energy benefits for the body
cytochrome
- an enzyme responsible for the transfer of electrons from NAD H to O 2. This process provides a significant energy benefit to the body.

Aerobic organisms most often contain three cytochromes, facultative anaerobes - one or two, obligate anaerobes do not contain cytochromes.

Anaerobic microorganisms can actively influence the environment, creating a suitable redox potential of the environment (eg Cl.perfringens). Some seeded cultures of anaerobic microorganisms, before starting to multiply, lower pH 2 0 from a value to , protecting themselves with a reductive barrier, others - aerotolerant ones - produce hydrogen peroxide during their vital activity, increasing pH 2 0 .

At the same time, glycolysis is characteristic only for anaerobes, which, depending on the final reaction products, is divided into several types of fermentation:

lactic acid fermentation Lactobacillus
,Streptococcus
, Bifidobacterium
, as well as some tissues of multicellular animals and humans.
alcoholic fermentation - saccharomycetes, candida (organisms of the kingdom of fungi)
formic acid - a family of enterobacteria
butyric - some types of clostridia
propionic acid - propionobacteria (for example, Propionibacterium acnes
)
fermentation with the release of molecular hydrogen - some types of Clostridium, Stickland fermentation
methane fermentation - for example, Methanobacterium

As a result of the breakdown of glucose, 2 molecules are consumed, and 4 molecules of ATP are synthesized. Thus, the total ATP yield is 2 ATP molecules and 2 NAD·H 2 molecules. The pyruvate obtained during the reaction is utilized by the cell in different ways, depending on what type of fermentation it follows.

Classification division of prokaryotes

The species diversity of these non-nuclear ones is enormous: science has described only 10,000 species, and there are supposedly more than a million species of bacteria. Their classification is extremely complex and is carried out based on the commonality of the following features and properties:

morphological - form, mode of movement, ability to spore formation, and others);
physiological - breathing with oxygen (aerobic) or an anoxic variant (anaerobic bacteria), according to the nature of metabolic products, and others;
biochemical;
similarity of genetic characteristics.

For example, the morphological classification by appearance subdivides all bacteria as:

rod-shaped;
winding;
spherical.

The physiological classification in relation to oxygen divides all prokaryotes into:

anaerobic - microorganisms whose respiration does not require the presence of free oxygen;
aerobic - microorganisms that need oxygen for their life.

Common growth media for anaerobic organisms

For general environment Wilson-Blair
the base is agar-agar with the addition of glucose, sodium sulfite and ferrous chloride. Clostridia form black colonies on this medium by reducing sulfite to sulfide anion, which combines with iron (II) cations to give a black salt.As a rule, black colony formations on this medium appear in the depth of the agar column.

Wednesday Kitta - Tarozzi
consists of meat-peptone broth, 0.5% glucose and pieces of liver or minced meat to absorb oxygen from the environment. Before sowing, the medium is heated in a boiling water bath for 20-30 minutes to remove air from the medium. After sowing, the nutrient medium is immediately filled with a layer of paraffin or paraffin oil to isolate it from oxygen access.

Cultivation of anaerobic organisms

Isolation of pure culture of anaerobes schematically

The cultivation of anaerobic organisms is mainly the task of microbiology.

For the cultivation of anaerobes, special methods are used, the essence of which is to remove air or replace it with a specialized gas mixture (or inert gases) in sealed thermostats. - anaerostats
.

Another way to grow anaerobes (most often microorganisms) on nutrient media is the addition of reducing substances (glucose, sodium formic acid, etc.), which reduce the redox potential.

Common growth media for anaerobic organisms

For general environment Wilson-Blair
the base is agar-agar with the addition of glucose, sodium sulfite and ferrous chloride. Clostridia form black colonies on this medium by reducing sulfite to sulfide anion, which combines with iron (II) cations to give a black salt. As a rule, black colony formations on this medium appear in the depth of the agar column.

Prevention

What will be the result of the treatment? This largely depends on the type of pathogen, the location of the focus of infection, timely diagnosis and the right treatment. Doctors usually give a cautious but favorable prognosis for such diseases. In the advanced stages of the disease, with a high degree of probability, we can talk about the death of the patient.

Next article.

Anaerobic bacteria are those that, unlike aerobic bacteria, are able to survive and grow in an environment with little or no oxygen. Many of these microorganisms live on the mucous membranes (in the mouth, in the vagina) and in the human intestine, becoming the cause of infection when tissues are damaged.

Sinusitis, mouth infections, acne, otitis media, gangrene, and abscesses are some of the best-known diseases and conditions that such bacteria lead to. They can also enter from the outside through a wound or when eating contaminated food, causing such terrible diseases as botulism,. But in addition to harm, some species benefit a person, for example, by turning toxic vegetable sugars into useful ones for fermentation in the colon. Also, anaerobic bacteria, along with aerobic ones, play an important role in the ecosystem, taking part in the decomposition of the remains of living beings, but not as big as mushrooms in this regard.

Differential diagnostic nutrient media

environments gissa
("variegated row")
Wednesday Ressel
(Russell)
Wednesday Ploskireva
or baktoagar "Zh"
Bismuth Sulfite Agar

Ressel Wednesday
(Russell) is used to study the biochemical properties of enterobacteria (Shigella, Salmonella). Contains nutrient agar-agar, lactose, glucose and indicator (bromothymol blue). The color of the medium is grassy green. Usually prepared in 5 ml test tubes with a beveled surface. Sowing is carried out by an injection into the depth of the column and a stroke along the beveled surface.

Bismuth Sulfite Agar
It is designed to isolate Salmonella in its pure form from infected material. Contains tryptic digest, glucose, salmonella growth factors, brilliant green and agar. The differential properties of the medium are based on the ability of Salmonella to produce hydrogen sulfide, on their resistance to the presence of sulfide, brilliant green and bismuth citrate. Colonies are marked in black color of bismuth sulfide (the technique is similar to the medium Wilson-Blair
).