Campy, Helicobacter, Aeromonas, Vibrio Updated PDF

CAMPYLOBACTER, HELICOBACTER, AEROMONAS, VIBRIO, & PLESIOMONAS HANNAH REILLY MS, MLS, PA (ASCP) CAMPYLOBACTER AND HELICOBACTER MICROAEROPHILIC SPECIES WE WILL DISCUSS: SLENDER, CURVED GRAM-NEGATIVE CAMPYLOBAC...

CAMPYLOBACTER, HELICOBACTER, AEROMONAS, VIBRIO, & PLESIOMONAS HANNAH REILLY MS, MLS, PA (ASCP) CAMPYLOBACTER AND HELICOBACTER MICROAEROPHILIC SPECIES WE WILL DISCUSS: SLENDER, CURVED GRAM-NEGATIVE CAMPYLOBACTER JEJUNI RODS CAMPYLOBACTER FETUS OXIDASE POSITIVE HELICOBACTER PYLORI CATALASE POSITIVE MOTILE BACTERIA OF MEDICAL CAMPY oxidase (+)  CAMPYLOBACTER SMALL, CURVED, MOTILE, GRAM-NEGATIVE BACILLI WITH A REQUIREMENT FOR A MICROAEROPHILIC ENVIRONMENT; CANNOT BE ISOLATED USING ROUTINE METHODS; MOST ARE UREASE NEGATIVE CAMPYLOBACTER SMALL (0.2-0.5 ΜM BY 0.5- 5ΜM), SLENDER, CURVED, GRAM-NEGATIVE RODS WHEN TWO OR MORE CELLS ARE NEAR EACH chaining OTHER THEY APPEAR AS "S", OR SEAGULL SHAPES OR AS SPIRALS CHAINING OF INDIVIDUAL ORGANISMS CAN BE MISTAKEN FOR Campylobacter jejuni SPIROCHETES MOTILE WITH DARTING OR CORKSCREW MOTION CLINICAL SIGNIFICANCE OF CAMPYLOBACTER JEJUNI A PRIMARY INTESTINAL PATHOGEN IS #1 CAUSE OF GI DISTURBANCE IN THE WORLD HARBORED BY MANY DOMESTIC ANIMALS, PARTICULARLY POULTRY AND CATTLE TRANSMITTED BY FECAL CONTAMINATION OF WATER, MEAT, DAIRY, OR OTHER ANIMAL PRODUCTS SEASONAL – MOST CASES ARE IN THE SUMMER/FALL MOST COMMON IN CHILDREN AND YOUNG ADULTS INFECTION IS MARKED BY ABDOMINAL PAIN & CRAMPING, DIARRHEA, FEVER, AND CHILLS USUALLY RESOLVES IN 3-7 DAYS IF TREATED, ORAL ERYTHROMYCIN IS THE DRUG OF CHOICE CLINICAL SIGNIFICANCE OF CAMPYLOBACTER FETUS CAUSES DIARRHEA CAUSES SEPSIS IN STOOL, MANY LABS DO NOT FIND IT COST-EFFECTIVE TO DIFFERENTIATE BETWEEN JEJUNI AND FETUS. IF FOUND IN BLOOD, YOU WOULD SPECIATE DETECTING CAMPYLOBACTER PART OF ROUTINE STOOL CULTURE, SINCE SUCH A COMMON PATHOGEN NEED SPECIAL MEDIA SPECIAL TEMPERATURE MINIMAL NUMBER OF CONFIRMATORY TESTS  CAMPYLOBACTER MEDIA CAMPY BLOOD AGAR ENRICHED CONTAINS ANTIBIOTICS TO SELECT FOR CAMPYLOBACTER PLATED AS PART OF STOOL CULTURE SKIRROW BLOOD AGAR COLUMBIA AGAR BASE WITH ANTIBIOTICS AND SHEEP BLOOD BUTZLER BLOOD AGAR CAMPY BLOOD PLATE INCUBATION MICROAEROPHILIC ENVIRONMENT 5-10% O , 8-10% CO 2 2 AMBIENT AIR IS 21% O 2 AND 0.03% CO2 USE JAR WITH CAMPY GAS GENERATING ENVELOPE OR CAMPY BIOBAG 42 C INCUBATOR THIS TEMP. CUTS DOWN THE NORMAL FLORA  BIOCHEMICAL CHARACTERISICS FLAT, MOIST OR SHINY, GRAY SPREADY COLONIES ON CAMPY MEDIA OXIDASE POSITIVE CATALASE POSITIVE CURVED, “GULL WING”, OR S SHAPED GRAM NEGATIVE RODS PALE STAINING HAS “DARTING” MOTILITY ON WET MOUNT WILL NOT GROW IN ROOM AIR OR CO2 INCUBATOR CAMPYLOBACTER JEJUNI HIPPURATE POSITIVE NALIDIXIC ACID SENSITIVE CEPHALOTHIN RESISTANT  HELICOBACTER CURVED, MICROAEROPHILIC, GRAM-NEGATIVE RODS, MOST HAVING STRONG UREASE ACTIVITY HELICOBACTER HELICAL, CURVED OR STRAIGHT, UNBRANCHED GRAM-NEGATIVE RODS DARTING MOTILITY MICROAEROPHILES OXIDASE POSITIVE MANY SPP. ARE ALSO CATALASE POSITIVE UREASE ACTIVITY IS SPECIES DEPENDENT CLINICAL SIGNIFICANCE OF HELICOBACTER PYLORI CAUSES GASTRIC (PEPTIC) ULCERS AND GASTRITIS ESTIMATED TO BE IN 20%-40% OF ADULTS IN US MAY PLAY A ROLE IN GASTRIC CARCINOMA (STOMACH CANCER) INVADES GASTRIC MUCOSA ULCERS RESOLVE WHEN PATIENT IS TREATED WITH ANTIBIOTICS BIOCHEMICAL CHARACTERISTICS OXIDASE POSITIVE CATALASE POSITIVE UREA POSITIVE FROM GASTRIC BIOPSIES ONLY HELICOBACTER COLONIZES THE MUCOUS LAYER OF THE ANTRUM AND FUNDUS OF THE STOMACH BUT DOES NOT INVADE THE EPITHELIUM VIRULENCE DETERMINENTS: UREASE: NEUTRALIZES HCL WITH PRODUCTION OF NH3 ; AMMONIA MAY ALSO DAMAGE CELLS ADHESINS AND A VACUOLATING CYTOTOXIN FLAGELLA ENABLE IT TO BURROW THROUGH THE MUCUS LAYER HELICOBACTER IDENTIFYING H. PYLORI CAN BE CULTURED, BUT ISN’T OFTEN DONE SPECIMEN IS GASTRIC/DUODENAL TISSUE BIOPSY PERFORM A RAPID UREA TEST PLACE TISSUE IN UREA SUBSTRATE FOR 1-2 HOURS H. PYLORI HAS UREASE ENZYME, WILL CAUSE COLOR CHANGE AS IT CONVERTS UREA TO AMMONIA.  H. PYLORI BIOPSY STAIN STAIN BLACK BECAUSE THEY ARE IMPREGNATED WITH SILVER AND ARE THEREFORE EASILY DETECTABLE IN BIOPSY SPECIMENS. NUMEROUS BACTERIA CAN BE SEEN ON THE SURFACE OF THE ANTIBODY TESTING FOR H. PYLORI A BLOOD TEST CHECKS TO SEE WHETHER YOUR BODY HAS MADE ANTIBODIES TO H. PYLORI BACTERIA. IF YOU HAVE ANTIBODIES TO H. PYLORI IN YOUR BLOOD, IT MEANS YOU EITHER ARE CURRENTLY INFECTED OR HAVE BEEN INFECTED IN THE PAST. UREA BREATH TEST FOR H. PYLORI A UREA BREATH TEST CHECKS TO SEE IF YOU HAVE H. PYLORI BACTERIA IN YOUR STOMACH. THIS TEST CAN SHOW IF YOU HAVE AN H. PYLORI INFECTION. IT CAN ALSO BE USED TO SEE IF TREATMENT HAS WORKED TO GET RID OF H. PYLORI.  A STOOL ANTIGEN TEST CHECKS TO SEE IF STOOL ANTIGEN TESTING FOR SUBSTANCES THAT TRIGGER THE IMMUNE SYSTEM TO H. PYLORI FIGHT AN H. PYLORI INFECTION (H. PYLORI ANTIGENS ) ARE PRESENT IN YOUR FECES (STOOL). STOOL ANTIGEN TESTING MAY BE DONE TO HELP SUPPORT A DIAGNOSIS OF H. PYLORI INFECTION OR TO FIND OUT WHETHER TREATMENT FOR AN H. PYLORI INFECTION HAS BEEN SUCCESSFUL. AEROMONAS, VIBRIO, AND PLESIOMONAS GLUCOSE FERMENTERS, MAY ALSO SPECIES WE WILL DISCUSS: FERMENT LACTOSE/SUCROSE AEROMONAS HYDROPHILIA TSI HAS YELLOW BUTT VIBRIO CHOLERA OXIDASE POSITIVE VIBRIO PARAHEMOLYTICUS CATALASE POSITIVE VIBRIO VULNIFICUS MOTILE PLESIOMONAS SHIGELLOIDES GRAM NEGATIVE RODS CLINICAL SIGNIFICANCE OF AEROMONAS FOUND IN FRESH AND SALT WATER WOUND INFECTIONS WILL HAVE A HISTORY OF WATER EXPOSURE IS AN OPPORTUNIST, TENDS TO INFECT PEOPLE WITH HEPATIC DISEASE OR NEOPLASMS OPPORTUNISTIC INFECTIONS INCLUDE: GASTROENTERITIS UTI MENINGITIS EAR INFECTIONS AEROMONAS HYDROPHILIA MOST COMMON HUMAN ISOLATE LOVES WATER (HENCE IT’S NAME) NORMAL FLORA OF LEECHES, OYSTERS, FISH, FROGS, ALLIGATORS SINCE IT IS FOUND IN FRESHWATER, FAUCETS, SHOWERHEADS, AND SINK TRAPS CAN HARBOR IT IDENTIFYING AEROMONAS IS RECOVERED ON NORMAL MICROBIOLOGY MEDIA FROM BLOOD, WOUNDS, STOOL SCREEN STOOL BLOOD PLATE FOR OXIDASE POSITIVE COLONIES TO DETECT THESE TSI = A/A OXIDASE + INDOLE + BETA HEMOLYTIC, GREENISH COLONIES USUALLY A NON-FERMENTER ON MACCONKEY STRAIGHT GRAM NEGATIVE ROD (CAN BE CURVED) MOTILE BACTERIA OF MEDICAL VIBRIO oxidase (-) oxidase (+) GRAM STAIN VIBRIO Vibrio cholerae, the causative agent of cholera GENUS VIBRIO >30 SPECIES PRESENTLY DEFINED WITH 13 IMPLICATED IN FISH AND SHELFISH INFECTIONS, 1 IN POULTRY DISEASE, 12 IN DISEASE IN HUMANS ALL ARE OXIDASE-POSITIVE, GLUCOSE-FERMENTING, GRAM- NEGATIVE BACILLI THAT GROW ON BAP & MAC AGAR OXIDASE REACTION DIFFERENTIATES THEM FROM THE ENTEROBACTERIACEAE WHICH ARE ALL OXIDASE NEGATIVE MOST IMPORTANT MEMBERS FOR HUMAN INFECTIONS: VIBRIO CHOLERAE - ONLY ONE TO SURVIVE IN FRESH WATER; THUS MOST LIKELY TO CONTAMINATE DRINKING WATER VIBRIO PARAHAEMOLYTICUS - REQUIRES NACL VIBRIO VULNIFICUS - REQUIRES NACL PRIMARY HABITAT BRACKISH OR MARINE WATER 50 YEARS OF DATA ON CLIMATE AND POPULATIONS OF OCEAN MICROBES ARE BEHIND A WARNING THAT WARMING OCEANS CAUSED BY CLIMATE CHANGE MAY BE LEADING TO AN INCREASE IN CHOLERA AND OTHER INFECTIONS CAUSED BY VIBRIO BACTERIA TRANSMISSION TO HUMANS IS FECAL-ORAL INGESTION OF CONTAMINATED SEAFOOD OR WATER EXPOSURE OF OPEN WOUND AND MUCOSAL SURFACES TO CONTAMINATED WATER (E.G., SWIMMERS, FISHERMEN OR DURING FLOODS) CULTURAL FEATURES GROW ON A VARIETY OF SIMPLE MEDIA WITHIN A BROAD TEMPERATURE RANGE (14º TO 40ºC) GROW ON MAC AND BAP; USE BIOCHEMICAL TESTS TO DIFFERENTIATE BETWEEN THE VARIOUS VIBRIO SPP. USE SPECIAL MEDIUM TO ENHANCE ISOLATION AND FOR IDENTIFICATION MOST ARE HALOPHILES (REQUIRE NACL TO GROW) EXCEPT FOR V. CHOLERAE TOLERATE A WIDE PH RANGE (6.5 TO 9.0) BUT ARE SUSCEPTIBLE TO STOMACH ACIDS IF GASTRIC ACIDS ARE REDUCED, PATIENTS REQUIRE LOWER DOSES OF ORGANISMS TO ESTABLISH AN INFECTION THIOSULFATE-CITRATE-BILE SALTS-SUCROSE (TCBS) MEDIUM CAN ENHANCE ISOLATION VIBRIO CHOLERAE FORMS YELLOW COLONIES ON TCBS AGAR BOTH V. PARAHAEMOLYTICUS AND V. VULNIFICUS FORM GREEN COLONIES ON TCBS CLINICAL SIGNIFICANCE OF VIBRIO CHOLERAE A SALT WATER ORGANISM CAUSES CHOLERA, A SEVERE DIARRHEA CHOLERA EPIDEMICS STILL OCCUR WORLDWIDE TRANSMITTED THRU FECALLY CONTAMINATED WATER OUTBREAKS ARE ASSOCIATED WITH WARM SEASONS AND POOR SANITATION CONTAMINATED UNDERCOOKED SEAFOOD AND FISH ARE THE MOST COMMON FOOD SOURCES PRIMARILY LINKED TO DEVELOPING COUNTRIES, CAN BE FOUND IN UNITED STATES BECAUSE OF PEOPLE TRAVELING  CHOLERA CHOLERA HAS RAPID ONSET (3-10 HOURS) PROFUSE DIARRHEA, WITH “RICE WATER” STOOLS (LIQUID WITH FLECKS OF PUS) CAN CAUSE DEATH IF SEVERE ELECTROLYTE LOSSES AND DEHYDRATION OCCUR TREATED BY IV OR ORAL REHYDRATION THERAPY IF ISOLATED, NEEDS SENT TO DEPARTMENT OF HEALTH FOR SEROTYPING  CHOLERA COPIOUS, WATERY DIARRHEA COMPOSED OF FLUIDS AND MUCOUS FLECKS BUT NO BLOOD OR INFLAMMATORY CELLS = "RICE WATER STOOLS" DUE TO ACTION OF CHOLERA TOXIN MOST INGESTED VIBRIO CELLS DIE IN THE STOMACH, THUS REQUIRE A HIGH INOCULUM, AT LEAST 108 CELLS, TO INITIATE DISEASE rice-water stool in cholera VIBRIO CHOLERA: VIRULENCE FACTORS POLYSACCHARIDE CAPSULE LPS SIDEROPHORES PILI CHOLERA TOXIN ENTEROTOXIN PRODUCTION IS CENTRAL TO THE PATHOGENESIS OF CHOLERA BUT THE ORGANISM ALSO REQUIRES ADDITIONAL VIRULENCE FACTORS SO IT CAN COLONIZE THE SMALL INTESTINE AND ADHERE TO THE MUCOSAL CELLS. THE ORGANISM IS NOT INVASIVE VIBRIO CHOLERA HAS LIPOPOLYSACHARIDE FORMED OF O ANTIGEN, LIPID A AND CORE POLYSACCHARIDE ALSO HAS FLAGELLAR (H) ANTIGENS BUT ONLY O ANTIGEN IS USEFUL TO DISTINGUISH STRAINS THAT CAUSE EPIDEMICS MORE THAN 140 SEROGROUPS OF V. CHOLERAE (O1 TO O140) V. CHOLERAE O1 STRAINS ARE ASSOCIATED WITH EPIDEMIC CHOLERA NON-O1 STRAINS OF V. CHOLERAE USUALLY CAUSE SPORADIC CASES OF CHOLERA-LIKE ILLNESSES VIBRIO CHOLERA TWO SEROGROUPS OF THE SPECIES ALMOST ALWAYS PRODUCE CHOLERA TOXIN AND THUS CAN CAUSE EPIDEMIC DISEASE VIBRIO CHOLERA: O1 (THERE ARE TWO BIOTYPES OF O1: CLASSIC AND EL TOR) CLASSIC BIOTYPE CAUSED 6TH WORLDWIDE PANDEMIC AND EL TOR THE 7TH THE EL TOR STRAIN CAN SURVIVE IN WATER LONGER THAN THE CLASSIC, PRODUCES HEMOLYSINS AND CAUSES HIGHER CARRIAGE RATES VIBRIO CHOLERA: O139 (BENGAL) IS A NEW PANDEMIC STRAIN THAT AROSE IN INDIA IN 1992 AND IS SPREADING ACROSS ASIA; IT IS THE FIRST NON-O1 STRAIN CAPABLE OF CAUSING EPIDEMIC DISEASE  2 STRAINS OF V. CHOLERAE CLASSIC: CAUSED WORLDWIDE PANDEMICS OF PAST VERY VIRULENT IS STILL AROUND ESP. IN INDIA/BANGLADESH. EL-TOR: NEWER STRAIN CAUSES MANY CURRENT EPIDEMICS LESS VIRULENT BUT SPREADS EASILY AND PERSISTS IN ENVIRONMENT BECAUSE IT CAN LIVE IN ALGAE AND RIDE AROUND THE WORLD! CHOLERA TOXIN A LYSOGENIZED BACTERIOPHAGE ENCODES THE GENES FOR THE TWO SUBUNITS OF THE CHOLERA TOXIN THE AB EXOTOXIN ENTERS THE INTESTINAL EPITHELIAL CELL, ADP- RIBOSYLATES A MEMBRANE-BOUND GS PROTEIN TO TURN IT ALWAYS ON TO ACTIVATE ADENYLATE CYCLASE TO CREATE CAMP FROM ATP THE INCREASE OF INTRACELLULAR CAMP CAUSES ACTIVE SECRETION OF IONS AND WATER: CHOLERA IS A SECRETORY DIARRHEA Action of cholera toxin in intestinal epithelial cells VIBRIO PARAHEMOLYTICUS MOST COMMON TYPE ISOLATED IS HALOPHILIC (SALT-LOVING) A MEMBER OF THE NONCHOLERA VIBRIO SPECIES CAUSES GASTROENTERITIS AS A RESULT OF INGESTING CONTAMINATED UNDERCOOKED SEAFOOD, ESPECIALLY OYSTERS USUALLY FOUND IN COASTAL REGIONS DIARRHEA TENDS TO BE SELF-LIMITING, BUT IS MORE SEVERE IF PATIENT IS IMMUNOCOMPROMISED (CIRRHOSIS OR LIVER DISEASE) VIBRIO VULNIFICUS STRICT HALOPHILE (SALT-LOVING) THAT REQUIRES HIGH SALT CONCENTRATION IN MEDIA TO ISOLATE MOST COMMON NONCHOLERA VIBRIO SPECIES ISOLATED AFTER HURRICANE KATRINA PERSONS WITH LIVER DISEASE AND OTHER IMMUNOCOMPROMISING CONDITIONS ARE ESPECIALLY SUSCEPTIBLE TO TISSUE INFECTION AND SEPTICEMIA EXPOSURE FROM OPEN WOUNDS IN SALT/BRACKISH WATER OR INGESTION OF CONTAMINATED SEAFOOD CAUSES FULMINANT (SUDDEN AND VIOLENT) WOUND INFECTIONS AND SEPSIS REQUIRES IRON AS A GROWTH FACTOR: COMPROMISED PATIENTS WITH HIGH SERUM IRON LEVELS ARE PROVE TO INFECTION INFECTION FROM V. VULNIFICUS USUALLY ACQUIRED BY EATING RAW OYSTERS OR CONTAMINATION OF OPEN WOUND BY OCEAN WATER SYMPTOMS APPEAR RAPIDLY (POSSIBILITY OF DEATH W/IN 48 HOURS) TISSUE INFECTIONS PRESENT INITIALLY WITH FEVER, SWELLING, AND INTENSE PAIN AT SITE USUALLY LOCATED ON EXTREMITIES (HANDS, ARMS, FEET, LEGS) WITHIN HOURS HEMORRHAGIC BULLAE (BLOOD-FILLED BLACK BLISTERS) AND TISSUE NECROSIS APPEAR THE NECROTIC TISSUE CAN DEVELOP INTO NECROTIZING FASCIITIS AND RAPIDLY LEAD TO SECONDARY SEPTICEMIA IDENTIFYING VIBRIO OFTEN REQUIRES SPECIAL MEDIA: TCBS AGAR (THIOSULFATE CITRATE BILE SUCROSE) DR. SHOULD COMMUNICATE WITH LAB IF VIBRIO INFECTION IS SUSPECTED IF YOU WORK IN A COASTAL AREA, THIS MEDIA MAY BE INCLUDED ROUTINELY ON STOOL CULTURE PEPTONE WATER IS ALSO USED TO ENHANCE RECOVERY OF VIBRIO TCBS MEDIA IS A SELECTIVE AND DIFFERENTIAL MEDIA: SUCROSE IS THE DIFFERENTIAL ELEMENT SUCROSE FERMENTERS WILL HAVE YELLOW COLONIES NON-SUCROSE FERMENTERS WILL HAVE GREEN COLONIES V. CHOLERAE = YELLOW COLONIES V. PARAHEMOLYTICUS AND VULNIFICUS = GREEN COLONIES BIOCHEMICAL IDENTIFICATION OF VIBRIO GRAM STAIN: CURVED GRAM NEG RODS LOOK LIKE COMMAS OXIDASE + CATALASE + MOTILE MAY NEED TO ADD SALT TO BIOCHEMICAL TESTS TO GROW THE HALOPHILIC VIBRIOS BIOCHEMICAL REACTIONS V. cholerae V. parahemolyticus V. vulnificus Sucrose positive Sucrose negative Sucrose negative TSI: A/A TSI: K/A TSI: A/A Indole positive Indole positive Indole positive Oxidase positive Oxidase positive Oxidase positive Motile Motile Motile Does not grow in 6% salt Grows in 6% salt broth Grows in 6% salt broth broth TCBS Growth: yellow TCBS Growth: Blue/green TCBS Growth: Bue/green CLINICAL SIGNIFICANCE OF PLESIOMONAS SHIGELLOIDES CAUSES GASTROENTERITIS AND SEPSIS FRESH WATER ORGANISM LIKES WARMER CLIMATES ACQUIRED BY INGESTION OF CONTAMINATED FOOD OR WATER OR EXPOSURE TO REPTILES AND AMPHIBIAN FECES IS NOW INCLUDED IN THE ENTEROBACTERIACEAE FAMILY IDENTIFYING PLESIOMONAS IS RECOVERED ON NORMAL MICROBIOLOGY MEDIA FROM BLOOD, WOUNDS, STOOL STRAIGHT GRAM NEGATIVE ROD TSI = K/A OXIDASE + INDOLE POSITIVE OPAQUE PIGMENT USUALLY NON-LACTOSE FERMENTER ON MACCONKEY AGAR APPROXIMATELY 1/3 OF STRAINS MAY FERMENT LACTOSE GRAY NON-HEMOLYTIC COLONY ON BAP APPEARS GREEN (NONSUCROSE FERMENTER) ON HE AGAR WILL NOT GROW ON TCBS AGAR—HELPS DIFFERENTIATE FROM VIBRIO