Lecture 4

Symbiotic relations and microbes Normal microflore

Parasitic relations among microorganisms Bacteriophages

Symbiosis represents an intimate mutually relation of organisms of different species. They develop together better than separately. Sometimes the adaptation of two organisms becomes so great, that they lose their ability to exist separately (symbiosis of the fungus and blue-green algae, nitrogen-fixing bacteria.

Symbiotic relationship

Symbiosis of microorganisms and animals

Ants cultivate fungi in pure culture The fungi degrade cellulose that the animals cannot/

Anaerobic bacteria within the rumen degrade polyccharides

NORMAL HUMAN MICROFLORA

Normal microflora, or microbiota species that are generally found within the stable mixture of microbial community associated with particular body tissue is the result of a adaptation of microorganism to certain parts of the body 1.normal and constant microflora Resident floraMicrobes which can be repeatedly cultured from a given site over time

2.temporary (casual) microflora. Transient floraMicrobes which may initially inhabit a site after exposure, but do not persist
Colonization Establishment of a microbial population in the animal host

Skin flora
Coagulase-negative Staphylococci
S. epidermidis (opportunistic infections)

Skin "diphtheroids"
Corynebacterium species (opportunistic) Propionibacterium acnes (acne)

BODY SITE RESIDENT MICROBIOTA

Factors influencing composition

microbial

community

SKIN most abundant are non-pathogenic Staphylococcus, Sarcina; also occur Low water activity and fatty acids production Micrococcus, diphtheroids etc.; some limit numbers and types of fungi and Gram-negative bacteria microorganisms on the skin except in moist regions ( 80mln 1.2 bill.)

Patient 1
Anaerobes Aerobes Anaerobes

Patient 2
Aerobes

ORAL CAVITY

Suitable pH weakly alkaline and tC favourable medium, food substances, action of lysozyme

Gram smear of Throat Swab

Oral flora
"Viridans" streptococci (endocarditis)

160 species:

Streptococcus species (Gram positive cocci in chains)

S. mutans (dental caries) S. sanguis S. salivrius S. mitis S. sobrinus S. milleri group (brain, liver abscesses, bacteremia)

Actinomyces (Gram positive branching bacilli)

A. viscosus (dental caries) A. israelii (cervicofacial abscesses)

Anaerobic Gram negative bacilli (gingivitis, lung abscess)

Porphyromonas gingivalis Prevotella melaninogenicus Fusobacteria

Treponema denticola (spirochaete) (gingivitis)

Dental caries
Major etiological agent Streptococcus mutans Disease loss of tooth enamel, dentin abscess formation, loss of tooth Pathogenesis Formation of dental plaque (biofilm), calculus (deposition of salts) Bacterial biofilm, numerous species Production of extracellular polysaccharide polymers

Fermentation of sucrose, production of organic acids Demineralization of enamel, dentin

http://www.dent.umich.edu/research/loeschelabs/c

Factors influencing microbial Normal Bacterial Flora of the Skin Is Predominantly Anaerobic community composition Ability to resist nonspecific Upper respiratory tract (nasal cavity defenses salivas Skin Cultures from Forearm and nasopharynx) mostly Streptococci bactericidal features Aerobic (St. pyogenes, St. pneumoniae); also occur (lysozymes), mucus Neisseria, Haemophilus, Bacteroids, adsorption, phagocytosis by Staphylococci 1 leucocytes Subject Anaerobic
Lower respiratory tract none alveoli and bronchi are usually sterile Phagocytic Aerobic cells prevent colonization

Subject 2 Anaerobic

BODY SITE RESIDENT MICROBIOTA

Factors influencing microbial community composition

Vaginal tract Lactobacilli (Doderlein rods), Fusobacterium, Staphylococci, Streptococci, Bacteroids, yeast etc.

Large surface area and secretions of nutrients; acidity and bactericidal secrets limits species within population

Upper urinary tract (kidneys and bladder) none

Filtration and outward fluid flow prevent the establishment of microbiota

Normal vaginal Gram stain

Gram smear of Vaginal Swab

Bacterial Vaginosis
Pathogenesis: abnormal vaginal flora
association with absence of H2O2 producing Lactobacillus species (L. crispatus) elevated vaginal pH predominance of Gardnerella vaginalis
Gram variable coccobacilli (Gram positive structure)

overgrowth of anaerobic bacteria

Prevotella, Porphyromonas, Bacteroides, Atobobium vaginae, others

production of amines (trimethylamine) induces transudation and exfoliation of epithelial cells degradation of mucins producing thin homogenous discharge
Gram-negative anaerobic bacilli

increased production of cervical cytokines (IL1, IL8, TNF)

Bacterial Vaginosis
Diagnosis observation of "clue cell" on microscopic examination of vaginal smear ( Gram stain), adherent grayish discharge, production of volatile malodorous amines on addition of KOH vaginal pH > 4.5

"clue cell

BODY SITE RESIDENT MICROBIOTA

Factors influencing community composition

microbial

STOMACH mostly Lactobacteria, also occur Campylobacteria, Fungi, Sarcina and others. May be - Helicobacter pylori

Acidic pH, digestive enzymes limit the number of species pH-2,5 100 cells \ml pH-4,5 10000 cells\ml secretions,

SMALL INTESTINE Gall, mucus Lactobacteria, Enterococci, E. secretory IgA coli, Bifidobacteria etc. few species

Helicobacter pylori in Gastric Biopsy

LARGE INTESTINE (bowel) around 200 spp: I obligate non-spore-forming anaerobes (Bifidobacteria and Bacteroids) 96 99 %; II facultative anaerobes (E. coli, Lactobacilli, Enterococci); III very few Staphylococci, Proteus, Candida, Clostridium, Pseudomonas;

Abundance of substrates (pH, temp., humidity) for growth of resident microbiota;

About one-third of the dry weight of the faeces of is made up of microbes. Daily, an adult human excretes about 17 million billion micro-organisms with the excrements.

Gram smear Fecal Swab Gram smear of of Fecal Swab

LARGE INTESTINE
Representatives Quantity (in norm)in 1g
109-1011 (85-95%)

The eubiotic( probiotic) preparations

Bifidobacteria

Bifidobacterin Bificol

Lactobacilli Enterobacteria Bacteroides (nonspore forming anaerobes) Cocci Clostridium Baccillus

107-109 107-109 109-1010

Lactobacterin Colibacterin

105-106 103-104 103-104

Bioflorin( Italy)

Bactisubtil

THE ROLE OF NORMAL MICROBIOTA

Factor of nonspecific defense of the body against infections with pathogens a.) colonizatory resistance,

b.) production of allelopathic substances (bacteriocins, fatty acids, lactic acid etc.) c.) synthesis of nutrients essential to the host (vitamins B1, B2, B6, B12, K etc.) d.) participation in metabolism of some compounds (cholesterol, gall acids, vitamins)

e.) stimulation of immune system by microbial antigens

Imbalance of microbiota (e. g. by antibiotics) could lead to development of disease (disbacteriosis, psudomembranous enterocolitis etc.) administration of normal microflora preparations probiotics: colibacterin lactobacterin bifidobacterin bificol acelact Normal microbiota under certain conditions could lead to the development of indigenous infections

Parasitism

(parasitos sponger)

Bdellovibrio

BACTERIOPHAGES
viruses whose host is a bacterial cell CLASSIFICATION OF PHAGES Origin Genome single-stranded double-stranded DNA RNA
Gr.1 DNA containing, lyze bacteria with F- plazmids Gr.2 small RNA containing Gr.3 - odd (3,7 ) T-phages with short tail Gr.4 odd (1,5 ) -phage tail is contractil Gr.5 even (2,4,6 ) DNA containing, tail is contractil

E. Coli, C. Diphteria, V. Cholera, etc.

F.DErrel -1916 year

Morphology

Cubic (isometric) - all kind of

genome

Filamentous (helical) -singlestranded DNA

Complex - long or short

contractile tail double-stranded DNA

Specificity (host cell range)

Polyvalent replicate within host

cell of several species

Monophages replicate within

specific host cell species

Typing phages replicate only

within specific host cell strain

Type of interaction with bacterial host 1.Lytic phage (virulent) 2.Temperate phage (lysogenic)

STAGES OF PHAGE REPLICATION Specific adsorption Penetration and uncoating Establishment of phage genome Phage proteins and NA synthesis Assembly Release

Andr Michel Lwoff 1902-1994 Nobel prize 1965 y.

APPLICATION OF PHAGES Therapy Prophylactic Typing, diagnostic Gene engineering DIAGNOSTIC Identification of certain bacteriophage in sample Identification of microorganism species in sample Phage typing identification of strain and source of the pathogen.

Practical application of bacteriophages

Treatmenf of inf. dys

1. Disenteria bacteriophages Polyvalent

Filtrate of bacteria lysate

2. Salmonella bacteriophages Polyvalent

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3. Salmonella typhy bacteriophages Staphylococcal bacteriophages Polyvalent

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