Gram-positive bacilli ( Spore forming, Non-Spore-forming ) 2008274004 임창하 자료조사&피피티제작 2010251114 김덕호 자료조사&피피티제작 2010274002 심예슬 자료조사&피피티제작 2010274007 이소윤 자료조사&피피티제작 2011274021 박혜란 자료조사&발표

Content Gram positive Spore-forming bacilli - Bacillus - Clostridium Gram positive Non-Spore-forming bacilli - Listeria - Erysipelothrix, - Corynebacterium - Propionibacterium

Gram positive bacilli 분류

Central, Sub-Terminal, and Terminal spores

The Genus Bacillus B.cereus : cause of one type of food poisoning Aerobic, Catalase(+), not fastidious Spore disperse by means of dust Versatility in degrading complex macromolecule common source of antibiotics Primary medical importance B.anthracis : cause of anthrax B.cereus : cause of one type of food poisoning

Bacillus anthracis Central spores Block-shape, angular Anthrax의 원인균 The largest of all bacteria pathogen Morphology 3~10μm long , 1~1.15μm width Characteristic Non-motile, Catalase + Central spores Block-shape, angular Virulence factor -polypeptide capsule -exotoxin

Anthrax Anthrax type Cutaneous(95% of human cases) hemorrhagic vesicle covered black crust

Anthrax 2. Pulmonary (100% fatal) capillary thrombosis cardiovascular shock 3. GIT/systemic (rare) vomiting, diarrhea, toxemia, death

Anthrax Treatment -penicillin, tetracycline -vaccination of animal herds and people in endemic areas is performed

Bacillus cereus flagella Central spore Morphology facultative anaerobe Characteristic β-hemolytic, Central spore Airborne & dust-borne contaminant Multiple readily in cooked food toxin-mediated food poisoning : fried rice, meat or vegetables

Bacillus cereus nauseam vomiting, abdominal cramps, diarrhea Symptoms no specific treatment symptoms diasppear within 24 hours Two toxins Short incubation period : vomiting (dairy product) Long incubation period : diarrhea (meat)

Bacillus subtilis Characteristic : Spore & flagella β-hemolytic (BAP) non-toxic non-toxic 비병원성

The Genus Clostridium Characteristic Anaerobic Catalase nagertive Endospore forming gram-positive bacilli Reside in a variety of habitats Infection occur when spores are introduced into injured skin 혐기성 카탈라아제 음성 > bacillus 속 아포형성 그람 양성 간균 여러곳에서 서식함 아포는 피부 상처를 통해 유입되 감염

Pathogens grouped according to toxin effect Neurotoxic C.botulinum : flaccid paralysis C.tetani : spastic paralysis Enterotoxic C.difficile : antibiotic associated diarrhea C.perfringens : gangrene, food poisoning 신경독소 이완성마비 강축성마비 2. 장독소 항생제 관련된 설사 괴저, 식중독

Clostridium perfringens found in soil, on human skin, intestine and vagina Non-motile Capsule Hemolysis on BAP-double zone Subterminal spore

Clostridium perfringens Gas gangrene - Damaged or dead tissue, Oxygen-poor internal muscles - anaerobic cellulitis , myonecrosis Produces toxin (lecithinase C) - red blood cell rupture, edema, tissue destruction Produces enzymes - collagenase, DNase, hyaluronidase => Pain, edema, bloody exudate, fever, tachycadia, blackned necrotic tissue 가스괴저 손상되거나 죽은 조직, 산소가 부족한 내부근육에서 발병 무산산소성 연조직염(봉와직염), 근괴사 독소를 생성 – 세포막에 구멍을 뚫음, 장->장관염, 적혈구 -> 용혈 => 적혈구 파괴, 부종, 조직파괴 효소를 생성 => 콜라겐분해효소, Dnase, 하이알루론산분해효소 통증 부종 피고름 열 심박급속증 흑색 괴사 조직

Gas gangrene

Treatment, Prevention surgical repair, amputation debridement of diseased tissue Hyperbaric oxygen therapy Immediate cleansing 잘라내는것이 가장 효과적 (수술적 제거나 절단) 고압산소 요법은 산소의 증가로 인해 혐기성 세균의 증식과 독소생산을 막아 감염의 심각성을 줄일수 있다. 예방은 즉각적이고 철저한 세척

Clostridium tetani C.Tetani is recovered from the wound in only 30% of acases Motile Terminal spore (drumstick) Strict anaerobe

Tetanus Tetanospamin – neurotoxin Symptoms Clenching of the jaw -> Extreme arching of back -> Flexion of the armsextension of the legs -> Dead 테타노스파민 운경신경말단까지 퍼짐-> 척수의 축삭 -> 척수신경에 결합(골격근의수축을억제하는) -> 신경전달 물질의 방출을 억제 증상 – 턱에 경련 -> 등이 아치형으로 휨 -> 팔이 굽고 다리가 곧게 펴짐 그리고 경련미소가 동반 -> 호흡근육마비 -> 사망

Treatment and prevention Throughly cleansing and removing Penicillin, tetracyclin, muscle relaxants Antitoxin therapy -tetanus immune globulin(TIG) -tetanus antitoxin(TAT) Vaccine : tetanus toxoid 이미 진행중이라면 감염조직을 세척및 제거 -> 테트라싸이클린,페니실린,근육이완제를 사용 의심가는 사람은 항독소 치료를 행한다 - 파상풍면역글로불린 -> 비싸고 효과가 좋다 - 파상풍 항독소 -> 알레르기 반응이 일어날수도 있다 예방은 백신을 통해서 할수 있는데 백신은 파상풍변이독소가 들어있다.

Clostridium botulinum Commonly inhabit soil, water, and intestinal tracts of animals Motile (flagella) β-hemolysis Botulism toxin (botulin) -Causes food poisoning Subterminal spore 토양과 물 동물의장관에 서식 운동성 있음 베타용혈성 보튤리즘 독소를 갖음(보튤린) 8개의 형이 존재 -> 빨강은 사람에게 감염 식중독의 원인 Subterminal spore

Botulism 1. foodborne botulism - enters the small intestine with food -> absorbed into the lymphatics -> neuromuscular junction of skeletal muscles -> prevent the release of acetylcholine-> most potent microbial toxin - improperly sterilized canned food 2. wound botulism - infected wound / inject heroin /symptoms similar to those of food-borne botulism 3. infant botulism - In children between the ages 2 weeks and 6 months 음식매개 보튤리눔독소증 음식과함께 섭취 -> 작은창자에 도달 -> 림프계와 순환계로 흡수 -> 골격근의 신경근 접합부에 도달 -> 아세틸콜린(근육수축) 방출을 막음 원인 : 살균이 안된 캔음식 증상 : 복시, 연하곤란, 현기증 -> 하행근육마비, 호흡계손상 2. 외상 보툴리눔독소증 파상풍처럼 외상을 통해 감염 하지만 증상은 식품매개 보튤리늄과 비슷하다 피부에 마약을 주입하는사람에게선 감염률이 증가한다. 3. 유아 보튤리눔독소증 2주~6개월 유아가 아포를 섭취하여 발생함 증상 : 이완성마비, 긴장저하영아증후군, 호흡합병증

Treatment and Prevention - type A, B, E trivalent horse antitoxins - respiratory and cardiac support systems - penicillin Prevention - home-preserved foods : preservatives (sodium nitrate, salt, acid) - toxin is heat-sensitive, rapidly inactivated at 100℃ boiled for 10mins before eating 치료 3개의 말 항독소 A B E 형을 투여 심호흡지지장치 페니실린 예방 보존제 첨가 (질산나트륨 소금 식초 독소는 열에 감수성이 강함 따라서 100도씨에서 10분간 가열해주면 빠르게 불활성화됨

Clostridium difficile flagella subterminal spore Produces A(enterotoxin) and B(cytotoxin) toxins CDAD ; Clostridium difficile associated disease 운동성 독소를 생성 항생제 관련 위막성 대장염

CDAD ; Clostridium difficile associated disease Antibiotic associated diarrhea Pseudomembranous colitis Treatment and Prevention vancomycin, metronidazole Vaccin : : C.difficile toxoid ELISA testing 항생제 관련 설사 위막성 대장염 엠피실린 클린다마이신 세파로스포린과 같은 광범위 항생제로 치료중에 생김 장독소 -> 창자벽 괴사(비침습성) -> 설사 -> 복부경련 열 백혈구증가 위막탈락 치료 – 반코마이신 메트로니다졸 경구 복용 예방 – 백신 –위독소 - ELISA를 통해 빠르게 감별 ELISA 항체나 항원에 효소를 표지하여 효소의 활성을 측정하여 항원-항체 반응의 강도와 그 양을 정량적으로 측정하는 방법 = 효소 결합 면역 침강 분석법

Diagnosis of clostridial species gas chromatography : - analyzes the chemical differences among species direct (ELISA) testing of isolates toxicity testing in mice or guinea pigs serotyping with antitoxin neutralization tests 클로스트리듐 종의 감별진단 가스크로마토그래피 – 종들간의 화학적 차이를 분석 -> 복잡함 분리주의 직접 ELISA 검사 쥐나 기니피그를 이용한 독성검사. 항독소 중화를 이용한 혈청형 검사

Listeria monocytogenes An Emerging food-borne pathogen Gram(+) Regular Non-spor-forming bacilli From cocobacilli to long filaments in palisades formation Tumbling motility with one to four flagella not capsules or spores Not fastidious, (resistant to cold, heat, salt, pH extremes, bile) β-hemolytic Unique multiplication cycle

1) Epidemiology & Pathology The primary reservoir (water, soil) Secondary sources of infection (plant materials, the intestines of healthy mammals, birds, fish, invertebrates)

Predisposing factor : weakened condition of host defenses in intestine mucosa In normal adults : mild of subclinical infection (fever, diarrhea, sore throat) In immunocompromised patients,fetuses,neonates : infection of the brain and meninges → septicemia Pregnant women : Transmitted to the infant Intrauterine infection → premature abortion, fetal death Neonatal infections → damage to the nervous system

2) Diagnosis Cold enrichment ELISA Immunofluorescence 3) Treatment Ampicillin, Erythromycin trimethoprim-sulfamethoxazole Cold storage : not an effective control measure

Erysipelothrix rhusiopathiae A zoonotic pathogen Gram(+) Regular Non-spor-forming bacilli 1)Epidemiology, Pathology Primary reservoir : tonsils of healthy pigs, sheep, chickens, fish In sewage, seawater, soils, foods Swine erysipelas

Penicillin, erythromycin Erysipeloid Swollen, inflamed, dark red lesions that burn and itch 2) Treatment Penicillin, erythromycin

Corynebacterium Gram-Positive Irregular Non-Spore-Forming Bacilli Motility x Spore x Stain → Metachromatic granule C.xerosis, C.striatum, C.diphtheriae

Corynebacterium diphtheriae Most human disease associated with C. diphtheriae Morphology 1. Pleomorphic variants 2. Metachromatic granule 3. Straight or somewhat curved rod, tapers at the ends 4. Club, filamentous and swollen shapes Pleomorphic : 형태학적 다양함 Palisade : 울타리 granule : 과립

Epidemiology of Diphtheria Significant cause of mortality but rate have steadily declined Most cases occur in non-immunized children living in crowded, unsanitary situations Still, many populations harbor a reservoir of healthy carriers

Pathology of Diphtheria Contact with droplets from human carriers or active infections Local infection (Cutaneous diphtheria) Toxin production & Toxemia Primary infection site : upper respiratory tract Cutaneous diphtheria starts from virulence factors that assist in its attachment and growth -> deep, erosive ulcers Not invasive but usually remain localized at portal of entry Erosive ulcer : 부식성 궤양 혈액 내에 침입하지 않고도 독소로 인해 전신증상을 나타낼 수 있다.

Diphtherotoxin and Toxemia - C. diphtheriae 가 감염부위에서 생성하는 매우 강력한 외독소 - Consisted of : Fragment A & B 1. Fragment A : interacts metabolically with factors in the cytoplasm and stops protein synthesis. 2. Fragment B : Binds to target cells in heart and nerve system 쉽게 말해 B가 target cell에 부착하면 A가 분리되고 cytoplasm으로 들어가서 독작용 Cardinal : 가장 중요한

Diphtherotoxin and Toxemia Local infection Inflammatory reaction, swelling neck and fever Systemic infection -Pseudomembrane Greenish-gray film, solidification of fluid in pharynx -Toxemia (독혈증) Toxin absorbed from throat carried by blood to heart and nerves -> Asphyxiation, Heart damage, Muscle weakness Pseudomembrane – 단단하게 붙어있어 떼려고 하면 출혈, 기도에 생겨 질식 유발 Myocarditis and abnormal EKG patterns : 심근염과 부정맥 Asphyxiation : 질식

Diagnosis for Corynebacterium Positive Schick test - Diphtheriae 변성 독소를 주입하여 반응 여부 검사 빨갛게 변함(+) : 백신 접종 필요 반응 없음(-) : 굳이 백신 접종할 필요가 없음 - 백신의 이차적인 부작용 결과를 최소화 디프테리아 변성독소(Toxoid)를 주입하여 체내에서 이 독소에 반응할 수 있는 능력이 있는지를 살핌. 빨갛게 변함(+) : 백신 접종이 필요함 아무 반응 없음(-) : 굳이 백신 접종을 할 필요가 없음. 디프테리아 변성독소에 양성반응을 보인 경우에만 항독소를 주입하는 백신을 접종하게 되었기에 백신의 이차적인 부작용 결과를 최소화 할 수 있게 되었다.

Treatment and Prevention of diphtheria - DAT (Diphtheria Anti-Toxin) : 말 혈청에 대한 allergy test필요 - Penicillin + erythromycin Prevention - DPT : Vaccination with toxoid - DTaP : DPT의 부작용을 최소한 한 Vaccine. - D : Diphtheria T : Tetanus by Clostridium tetani (파상풍) P : Pertussis by Bordetella pertussis (백일해) DAT : 말의 혈청이용, 말의 혈청에 대한 allergy test를 먼저 진행해야함. DTaP : D,T는 Toxoid, aP는 bacterial protein 성분. 항원 : Toxoid – 독작용 x 항원작용 o Bacterial protein 성분 포함

The Genus Propionibacterium Gram-Positive Irregular Non-Spore-Forming Bacilli Different by aerotolerant or anaerobic, nontoxigenic Propionibacterium acnes - Glands of human skin Genetic and hormonal factors Resemble Corynebacterium in morphology and arrangement P. acnes : 여드름 (화농성) 유발

QUIZ

In genus Clostridium, which make an Antibiotic-Associated Colitis? Clostridium difficile Clostridium tetani Clostridium perfringens Clostridium botulinum Clostridium novyi 답 1번

What is correct characteristic of Listeria monocytogens? Producing capsule and spore Growing in fastidious condition Causing premature abortion Not living in 4℃ Invading tonsil 답. 3번.

Which of the following is not C.diphtheriae’s morphological type? Curve rods Filamentous Club shape Lancet shaped Palisades arrangement 답. 4번.

Which is not used for treatment & prevention of C.diphtheriae? DTaP DAT Penicillin Erythromycin None of these 답. 4번.