klostridit 2011 eng

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Clostridia

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Clostridia

Large Grampositive anaerobic rods

Form of spores resistant to oxygen, drought,

freezing and cooking temperatures

The soil and in the intestines of warmblooded

Cause the food, the environment and the points of

through drug poisoning and infectious

the toxininfections

Page 3 A. Clostridium botulinum

1. The classic botulism

2. The infant botulism

3. The wound botulism

B. Clostridium perfringens

1. The classic food poisoning



2. enteritis necroticans

C. clostridium tetani

1. Tetanus

D. Clostridium difficile

1. Clostridium difficile assossioitunut diarrhea (CDAD)

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A. Clostridium botulinum

1. Classical botulism

Food poisoning, the risk of food products, especially meat, fish, vacuum packed

Products and preserved home products

Clostridium botulinum secretes proteiinieksotoksiinia, which is the strongest

known as toxic (lethal dose: 0.1 ng / kg)

Neurotoxins are synthesized as an inactive polypeptide, i.e.,

protoksiineina (size 150 kDa), which are activated when the protease cleaves

polypeptide chain

C. botulinum are classified into seven serotypes AG during growth

on the basis of the antigenic characteristics of their production of toxins types A,

B, E and F cause botulism people



A few other klostridilajit (C. butyricum, C. Barati) to produce

the same toxins as C. Botulinum

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Functional form of botulinum toxin can bind

nerve muscle joints and prevents the neurotransmitter

release of vesicles containing acetylcholine. Since

the muscle may no longer be any kind of messages to the nerve cells, muscle

paralyzed.

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the toxin is absorbed in the stomach and small intestine, from where it

spreads throughout the body, can also be absorbed by inhalation



Symptoms begin 1236 hours after ingestion of food

The first botulism symptoms include fatigue, weakness,

vomiting, then voice and difficulty in swallowing and visual disturbances

Respiratory depression can ultimately lead to death

Mortality of the disease is about 65%, in developed countries is 10%..

Risk group: dangerous for everyone

Even one person's illness botulism epidemic is interpreted

a serious disease Due to the nature of the

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2009

In early September, it was found in France in three at the same

the family of a person who is Clostridium botulinum

botulism caused by bacteria, the suspected source

Purchased from Finland to hotsmoked whitefish. Vacuumpacked fish

was purchased in Eastern Finland in retail trade in August

At the end, transported by plane to France

and by a private individual eaten two weeks



later on the last day of use. Journey, the fish

had been stored in a cool box

unknown temperature for 14 hours and then

After the home refrigerator. Before eating fish is not

was heated.

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2006

Clostridium botulinum caused in July 2006 for one

epidemic, which contracted by two persons of the same

family. The vehicles were

likely to be hotsmoked whitefish, which was purchased

packed in a local supermarket. Fishfinal

date was weeks away syömisajankohdasta. Fish

the country of origin was Canada, it was delivered to Finland

frozen. Defrosting and smoking, as well as the finished product

packaging took place in a domestic establishment.



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Older epidemics

1981 two German tourists got sick after eating

säilykemaksapasteijaa

1999 one person ill with whitefish roe

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Botulism treatment:

ABE trivalent antitoxin (given before symptoms

the start)

Gastrointestinal tract is cleared



Missed the gut bacteria are destroyed penicillin

Artificial respiration

Forecast the worse the sooner the symptoms appear

Recovery may take months

Waiting for new nervemuscle junctions formation

Small doses of purified botulinum toxin can be

use of CNS lihaston dysregulation,

in the treatment of diseases, the cosmetic use of biological

warfare

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2. The infant botulism

C. botulinum produces toxin in the baby's gastrointestinal tract (n. 80

spores / g)

Risk foods honey, vegetables and fruits

Less than half years more likely to get infant botulism;

immature intestinal flora, and a higher pH in the digestive tract

allow bacterial spore growth and yield of the toxin in the gut

Symptoms of constipation, muscle slackening, weeping sluggish

The symptoms are surprisingly mild, and mortality only a few percent,



causing sudden infant death syndrome

Power of treatment is usually rescued

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3. The wound botulism

C. botulinum spore's wound, where they germinate

The disease resembles tetanus

The use of injecting drugs

Cases detected in the Americas, Europe, rare

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Clostridium botulinum



• task analysis in mice

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Clostridium botulinum

PCR / PFGE / ELISA

M. Lindström Finland, studied the method:

käytettään primeriparia 4, which are

specific neurotoxins botulinum toxin A, B, E or



F. Four serotype simultaneous detection

possible.

ELISA ready for a day when the toxin in the sample

A viable cell growth and toxin

generating identification of genes by PCR

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B. Clostridium perfringens

1. The classic food poisoning

C. perfringens Type A enterotoxin secreted last

years, the most commonly reported cause of food poisoning

In Finland

The risk of foodmeat and fish, incompletely cooked,

slowly chilled, at too high a temperature, stored and

incompletely cooked foods

Causing disease dose of 10 ^ 6 to 10 ^ 8 cells

The incubation period of 824 h



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C. perfringens cells increase in food, some of the cells die of the stomach

acidity, but some cells remain alive in the small intestine, where

they begin to proliferate

When the bacterial cell itiöi, consists poisoning causing CPE

enterotoxin, if the position is cpe gene encoding it (only 5%

strains)

CPE toxin damage the surface of the epithelial cells of the small intestine

by making holes in the cell membranes to small molecules such as anions,

cations and organic acids leak out

Symptoms of abdominal pain, nausea and intense diarrhea that last for 1 to 2

days, the elderly C. perfingens can cause persistent diarrhea

Enterotoxin can be demonstrated in faeces

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2. Enteritis necroticans (pigbel)



Clostridium perfingens Type C secreted by β

enterotoxin can cause severe enteritis

necroticansin (necrotizing inflammation of the bowel)

The bacteria secrete a toxin from the food, which

lead to paralysis, haematomas and regional

necrosis, especially in the small intestine

Symptoms: vomiting, diarrhea, abdominal pain Hard,

hemorrhage

Rare

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Clostridium perfringens



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Clostridium perfringens plating

TSC Agar (Tryptose Sulfite

Cycloserine Agar)

to produce black colonies

cycloserine to prevent other bacteria

growth

In addition to kanamycin and polymyxin

inhibition of bacterial flora



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TSC

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Clostridium perfringens PCR and microarray



Microarray technology with antibodies

Identification of C. perfingens toxins

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C. Clostridium tetani

1. Tetanus, or lockjaw

C. tetani produces two poison, and tetanoslysiinia

tetanospasmiinia, of which the latter is said to

tetanus toxin

Tetanus neurotoxin, and the second strongest poison

After botulinum toxin

The bacteria can come into the wound, where it begins to increase

under anaerobic conditions



Infection is local, but the toxin spreads

via the bloodstream and lymph circulation, as well as directly to the

the nerve fibers along the spinal cord and brain stem

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toxin affects the central nervous system

hermosynapseihin and motor

end plates, blocks the inhibitory

neurotransmitters (e.g. glycine)

release, wherein the muscle

is prevented and the relaxation of the

muscle spasms

The incubation period of days to months

At the beginning of muscle spasms of the wound

close, spreads to the muscles of the jaw and

all parts of the voluntary muscles

Tetanus leads

if untreated, is almost always

death, manicured mortality

1030%

Prevented through vaccination

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Clostridium tetani



Page 26Page 27Page 28

C. tetani penetration



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Clostridium tetani determination

Genetically IgG ELISA IgG class are determined

antibodies

Mouse Test



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Clostridium tetani, the Mouse test

The left hind leg of the mouse injected

Tetanus toxin

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Clostridium difficile (40006000 cases / yr)



Insulation CDS (C. difficile

SELECTIVE) agar containing a Dcycloserinia and

Cefoksitinia

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Clostridium difficile

Clostridium difficile associated diarrhea

(CDAD)

Diagnostic tests for CDAD: in

the determination of the market. They

of toxin B (CytoTox assay)

or toxins A and B (immunoassay)

the genetic analysis.



Page 33Quick Test: Clostridium difficile Toxin A detection

of C. difficile toxin A in faeces with

Rapydtest®

10 15 min.



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Sources:

Austin, JW and Dodds KL 1997. Clostridium botulinum. In: Food Microbiology. ASM Press. Pages288304.Hielm, S. clostridia, the quiet. In: M. Salkinoja Salonen (ed.). Microbiological criteria. Microbiologypublications 49/2000.Microbiology Department of Applied Chemistry and Microbiology, University of Helsinki. Pages 652655.

SagittariusSomer, H. and Ristola, M. 2003. Clostridium species. In: P. Huovinen (ed.).Microbiology andinfectious diseases. Book 1. Duodecim Medical Publications. Pages 228236.

Madigan, MT 2003. Brock Biology of Microorganism. Pearson Educational.

Guide microbiological food and drinking water hazards. Evidential EELAOpas 1/2003. Pages 3744.

McClane BA in 1997. Clostridium perfringens. In: Food Microbiology. ASM Press. Pages 305326.

SalkinojaSalonen, M. clostridial toxins. In: M. Salkinoja Salonen (ed.). Microbiological criteria.Microbiology publications 49/2000.Division of Microbiology, Department of Applied Chemistry and Microbiology, HelsinkiUniversity. Pages 548549.

Pictures:

http://www.agen.ufl.edu/chyn/age2062/lect/lect_25/FG19_012.GIFhttp://wwwuser.gwdg.de/hbruegg/ct/tetanus2.gifhttp://www.agen.ufl.edu/chyn/age2062/lect/lect_25/FG19_011.GIF

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Fountains

Lindström, Miia; Keto, Riikka; Markkula, Annukka; Nevas, Mari; Hielm,

Sebastian; Korkeala, Hannu. Multiplex PCR Assay for Detection and

Identification of Clostridium botulinum toxin Types A, B, E, and ... & Applied



Environmental Microbiology, Dec 2001, Vol. 67 Issue 12, p5694, 6p, 4

charts, 1bw; (AN 6063605)

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