Non-Fermentative Gram-Negative Bacilli PDF

NON-FERMENTATIVE GRAM-NEGATIVE BACILLI Enterobacteriaceae. So, in terms of other general characteristics, they are also aerobic, non-spore-forming, Gram-negative bacilli...

NON-FERMENTATIVE GRAM-NEGATIVE BACILLI Enterobacteriaceae. So, in terms of other general characteristics, they are also aerobic, non-spore-forming, Gram-negative bacilli straight or slightly curved slender gram-negative bacilli. ü Aerobic Almost all are motile, except your Burkholderia mali. ü Obligate anaerobic Pseudomonas Aerobic gram-negative bacilli Gram-negative bacillus or coccobacillus ü Fermenters/Fermentative – ferment carbohydrates Strictly aerobic metabolism ü Non-fermenters – do not ferment carbohydrates Motile usually with polar flagellum or polar tuft of flagella Non-fermentative gram-negative bacilli/coccobacilli Oxidase positive (except P. luteolus and P. oryzihabitans) ubiquitous and found in most environments, typically Catalase positive in soil and water, on plants and decaying vegetation, Usually grows on MAC agar and in many foodstuffs. Usually an oxidizer of carbohydrates, but some They prefer moist environments, and in hospitals, species are asaccharolytic they can be isolated from nebulizers, dialysate, fluids, saline, catheters, and other devices. Pseudomonas aeruginosa is the most commonly isolated Non-fermenters can even withstand treatment with species of the genus in clinical specimens. It is found in your chlorhexidine and your quaternary ammonium moist environments, pools, hot tubs, catheters, and compounds. humidifiers in hospitals. GENERAL CHARACTERISTICS Pseudomonas aeruginosa Ø aerobic, non-spore-forming, straight or slightly opportunistic pathogen curved, slender, gram-negative bacilli VIRULENCE FACTORS Ø Motile except Burkholderia mallei ü exotoxin A Ø Pseudomonas and Burkholderia are oxidase positive ü exoenzymes S and T Ø Oxidizers (K/K on TSI) ü proteolytic enzymes (e.g., elastases) ü hemolysins (e.g., phospholipase C) The members of this genera use a variety of carbohydrate, alcohol, and amino acid substrates as ü pili and adhesins carbon and energy sources. Some group members even ü Alginate ü Pyocyanin oxidize carbohydrates to derive energy for their metabolism. They are referred to as oxidizers. So when leading cause of nosocomial respiratory tract they say oxidizer on your TSI-A, they will present as K infections over K on your TSI. Bacteremia with ecthyma gangrenosum Wound infections To emphasize, remember that in your TSI, oxidizers Common cause of lung infections in people with CF present as K over K on your TSI, meaning alkaline slant Otitis externa (Jacuzzi or hot tub syndrome) over an alkaline butt. So usually, they derive their energy common cause of ventilator associated pneumonia, for their metabolism on oxidizing your carbohydrates. That's why they are referred to as oxidizers. Pseudomonas aeruginosa is an opportunistic pathogen with a lot of virulence factors that contribute to their Others do not break down carbohydrates at all. They are pathogenicity, including the production of exotoxin A, referred to as non-oxidizers or as acarolytic. Additional which kills host cells by inhibiting protein synthesis, characteristics that can differentiate this group of non- exoenzymes S and T that disrupts the cytoskeleton fermenters from other gram-negative bacilli is motility, organization, and production of several proteolytic pigmentation, and their ability or lack of ability to grow on enzymes such as elastases and hemolysins such as selective gram-negative media such as MacConkey Agar. phospholipase C that are capable of destroying cells and Most non-fermented gram-negative bacilli are oxidase tissue. positive, a feature that differentiates them from the PANANGGILAAN, J. On the bacterial cell surface, pili and adhesin mediate and infection of the external ear canal, which is also what we reinforce attachment to host cells. Some strains produce call as malignant otitis externa, which can even alginate, which is a polysaccharide polymer that inhibits progress to involve the underlying tissue of the bones of phagocytosis, contributing to the infectious potential in the skull. And aside from that, Pseudomonas aeruginosa your cystic fibrosis patients. This substance also protects is also a common cause of ventilator-associated the bacteria from dehydration and antibiotic activity. pneumonia, especially in hospitalized patients. Pyocyanin is the blue phenazine pigment that contributes Identifying Characteristics to the characteristic green color of Pseudomonas aeruginosa that damages cells by producing reactive Pyoverdin pigment - yellow-green or yellow-brown oxygen species. pigment Pyocyanin pigment - blue-green The reactive oxygen species are also bactericidal to the fruity, grapelike odor organism to protect itself and the organism produce denitrification of nitrates and nitrites catalase enzymes to prevent destruction. P. aeruginosa arginine dihydrolase (ADH)-positive also contains several genes involved in quorum sensing. growth at 42° C This is a mechanism for detecting bacterial products in the citrate positivity immediate environment. When the growth of organism or acetamide utilization neighboring bacteria reaches a critical mass, the concentration of these inducing molecules reaches a level Identifying characteristics of Pseudomonas are very that activates transcription of virulence factors, including distinct from other bacteria, especially in its pigment genes related to metabolic processes, enzyme production. So, most isolates of Pseudomonas production, and biofilm formation. aeruginosa are beta hemolytic on sheep blood agar and will produce flat spreading colonies with a characteristic Biofilm production related to the overproduction of metallic sheen. This metallic sheen may be due to your alginate and the mucoid phenotype is isolated from cystic pyroverdin pigment, which is a yellow-green or yellow- fibrosis patients is associated with serious infections. In brown pigment. addition, Pseudomonas aeruginosa can survive harsh environmental conditions and display intrinsic resistance Many strains also produce a fruity, grape-like odor caused to a wide variety of antimicrobial agents, two factors that by the presence of 2- aminoacetophenone. Aside from facilitate the organism's ability to survive in the hospital pyroverdin, there is another pigment produced by setting. Pseudomonas aeruginosa, and that is your pyocyanin, which is more common. It's a blue-green pigment that is Pseudomonas aeruginosa is the leading cause of also water-soluble and capable of fluorescence. nosocomial respiratory tract infections. It causes bacteremia with ecthyma gangrenosum. So, ecthyma Aside from that, your Pseudomonas aeruginosa is also gangrenosum is when the blood vessels of the capable of denitrification of nitrates and nitrites. It's subcutaneous develops this necrotic rash or papules that arginine dehydrolase or ADH positive. It's able to grow at happens during a bacteremia. 42 degrees Celsius. It's citrate positive, and it utilizes acetamide. It also is a pathogen isolated from wound infections and is common cause of lung infections in people with cystic Cetrimide agar fibrosis. Now, always remember that when Pseudomonas Ø selective and differential medium for the identification aeruginosa is isolated from a sterile body site, such as a of P.aeruginosa blood, pleural fluid, joint fluids, or tissues, or cerebrospinal Ø Your cetrimide acts as a detergent and inhibits most fluid, it almost always constitutes a true infection. And bacteria. then, otitis externa, this is in particular in swimmers or Ø The medium also enhances the production of the two divers, is a necrotizing rash referred to as jacuzzi or hot pigments produced by Pseudomonas aeruginosa. tub syndrome that develops in users of these recreational facilities. And infections of the nail beds in people with artificial nails. Now, patients with diabetes may suffer a severe PANANGGILAAN, J. OTHER PSEUDOMONAS oxidase and ADH positive but does not produce pyoverdin and is acetamide negative Pseudomonas fluorescens Pseudomonas pseudoalcaligenes and Pseudomonas Produce acid from xylose Pigment production alcaligenes negative Growth at 42°C negative Contaminants Positive gelatin hydrolysis Oxidase positive biochemically negative in many tests Pseudomonas putida Motile P. pseudoalcaligenes is ADH positive and willweakly Produce acid from xylose ferment fructose Pigment production negative Growth at 42°C negative Pseudomonas luteola and Pseudomonas oryzihabitans Negative gelatin hydrolysis Rare isolates but produces serious infection in Both fluorescence and putida are of very low virulence, humans especially co-infection with each other or with rarely causing clinical disease. They have been isolated other bacteria from respiratory specimens, contaminated blood Risk factors: presence of foreign materials (e.g., products, urethra, cosmetics, hospital equipment, and catheters), corticosteroid use, and fluids. Pseudomonas fluorescence and putida have been immunocompromised states documented, although rarely, as causes of UTIs, post- Differentiate by ONPG and esculin hydrolysis test surgical abscesses, emphysema, septic arthritis, and other wound infections. Pseudomonas putida has been Both Pseudomonas are gram-negative, non-fermentative, associated with catheter-related sepsis in patients with oxidase-negative bacilli that are catalase positive and cancer. motile and able to oxidize glucose, grow on MacConkey agar, and often produce an intracellular non-diffusible Isolation of Pseudomonas fluorescence from blood yellow pigment. culture bottles in asymptomatic patients has been responsible for clusters of pseudobacteria, probably Both species typically produce wrinkled or rough polyenes related to contaminated catheters and catheter-related at 48 hours. Notice the wrinkled yellow polyenes devices. Both species can grow at four degrees Celsius produced by Pseudomonas oryzae habitants on your and have been linked to associated septicemia. They also sheep blood agar at 48 hours. both produce acid from silos. Acinetobacter Both do not produce pigment and also are not able to grow at 42 degrees Celsius, unlike your Pseudomonas A. baumannii - the glucose-oxidizing, nonhemolytic strain aeruginosa. To distinguish the two, perform a gelatin A. Iwoffii - the glucose-negative, nonhemolytic strain hydrolysis where your Pseudomonas fluorescence is gelatin hydrolysis positive and your Pseudomonas putida A. haemolyticus. is gelatin hydrolysis negative. Associated with ventilators, humidifiers, catheters, and Pseudomonas stutzeri other devices wrinkled, leathery, adherent colonies that may Under Acinetobacter, we have three important, actually produce a light-yellow or brown pigment two, but sometimes we can also isolate Acinetobacter ADH negative and starch hydrolysis positive hemolyticus. But the most important are your Acinetobacter baumannii and your Acinetobacter lwoffii. Pseudomonas mendocina Acinetobacter baumannii is the glucose-oxidizing non- Contaminant hemolytic strain nonwrinkled, flat colonies that may appear with a A.lwoffii is the glucose-negative non-hemolytic strain. yellowish-brown pigment Now Acinetobacter infection is associated with ventilators, smooth buttery appearance PANANGGILAAN, J. humidifiers, catheters, and other devices. Colonization of It's associated with the following risk factors, central these equipments. venous catheterization, cystic fibrosis, residing in an ICU, malignancy, mechanical ventilation, neutropenia, Clinical Infections previous broad spectrum antibiotic therapy, recent surgery, and human immunodeficiency virus infection. primarily affect patients with weakened immune systems and coexisting diseases. Burkholderia cepacia A.baumannii: eye infection pneumonia in patients with CF or chronic granulomatous A.Iwoffii: contamination or colonization rather than disease (CGD) infection Wound infections resulting from the use of contaminated These infections usually occur in areas with a high fluid content, such as the urinary tract and respiratory tract, water. and in peritoneal fluids. Selective Media Now in terms of virulence factor, it's able to produce a Pseudomonas cepacia PC), OF base, polymyxin B, liposaccharide capsule, enabling it to survive in dry bacitracin, and lactose (OFBL) agar environments and adhere to mucosal epithelium via B.cepacia-selective agar fimbriae that contributes to the genus virulence. can produce a nonfluorescing yellow or green Identifying Characteristics pigment strictly aerobic, and they appear as gram-negative B.cepacia may lose viability on SBA or sheep blood agar coccobacilli or even gram-negative cocci on Gram in three to four days without transfers. BCSA is most stain effective on reducing overgrowth while maintaining good oxidase negative, catalase positive, and nonmotile recovery ofB.cepacia. It can also produce a non- Purple colored colonies on MAC fluorescing yellow or green pigment. Acinetobacter organisms can resist decolorization and The colonies are non- wrinkled and a trait that may be retain the crystal violet stain, leading to misidentification. used to differentiate isolates from Pseudomonas stutzeri If that happens, they can appear as gram-positive cocci in which also produces a yellow pigment. B.cepacia does smears made from blood culture bottles. If that happens, not fluoresce like your Pseudomonas aeruginosa but can perform an oxidase test where they are oxidase negative, produce a non-fluorescing yellow or green pigment that catalase positive, and even non-mortal. Also, they may may diffuse onto the media. produce purplish- colored colonies on MA. So remember The left colonies of B.cepacia are on your your Acinetobacter baumannii is saccharolytic and on the own oxidative-fermentative base. other hand, your Acinetobacter lwoffii is saccharolytic. So Polymyxin B, ambacitracin, and lactose it produces purplish colonies on MacConkey and so it may agar are yellow, if you can observe. be mistaken as an enteric or a which will distinguish it which is also oxidase negative. The results of your TSI will And on the right side is your colonies of reveal it's a saccharolytic, it's an oxidizer instead of a B.cepacia on your Pseudomonas cepacia fermenter. agar, which is where they are white surrounded by a pink-red pigment. Stenotrophomonas maltophilia Laboratory Identification very common in the hospital environment in debilitated patients and those who are Oxidase positive immunocompromised. Glucose oxidizers oxidase-negative, non-fermentative, gram-negative lysine decarboxylase and ONPG positive bacillus. ornithine decarboxylase negative bluish on MAC agar fail to reduce nitrate to nitrite positive for catalase, DNase, esculin and gelatin Motile hydrolysis, and lysine decarboxylase susceptible to SXT PANANGGILAAN, J. Burkholderia mallei Burkholderia gladioli Glanders resembles B. cepacia complex nonmotile, gram-negative coccobacilli found in patients with CF and CGD Glucose oxidizer yellow pigment after 48 to 72 hours of incubation Reduces nitrates to nitrites Motile ADH positive catalase and urease positive affects horses, mules, and donkeys, making it a grows on MAC agar, oxidizes glucose, is mannitol respiratory tract zoonotic infection. positive and decarboxylase negative AST method: broth microdilution with Brucella broth and incubation at 35° C in ambient air for 16 to 20 hours Burkholderia pseudomallei Melioidosis nonfermentative wrinkled colony Bipolar staining Ashdown medium Earthy odor B.pseudomallei is an aggressive granulomatous pulmonary disease caused by ingestion, inhalation, or inoculation of the organisms with metastatic abscess formation in lungs and other viscera. B.pseudomallei produces a wrinkled colony, just like your Pseudomonas stutzeri, but your Pseudomonas stutzeri does not use lactose in contrast to your Pseudomallei. It also stains bipolarly, and a selective medium known as ash down medium is supplemented with colistin. Colonies on this agar are deep pink because of the absorption of neutral red in the medium. Colonies will also exhibit an earthy odor. However, work should be done, remember, in a biological safety cabinet when Pseudomallei is suspected, and avoid directly inhaling, directly sniffing the plate, so avoid doing that. Okay, so notice the wrinkled colonies and even the pinkish colonies on your, this is MacConkey, no this is not MacConkey, this is on your sheep blood agar. PANANGGILAAN, J. GRAM NEGATIVE BACILLI Now, its primary virulence factor is known as cholera toxin or cholera gene. And to effectively release this toxin, the VIBRIO organism must first infiltrate and distribute itself along the gram-negative, facultative anaerobic, curved or cells lining the mucosal surface of the gastrointestinal comma-shaped rods. tract. Require sodium for growth and glucose fermentation Motility and chemotaxis mediate the pro-distribution of all species are motile and catalase and oxidase organisms and mucinase production allows penetration of positive except Vibrio metschnikovii the mucous layer. Toxin-co-regulated pili provide how EPIDEMIOLOGY bacilli attach to mucosal cells for the release of cholera toxin. Primary habitat is water The enterotoxin zona occludens toxin has been shown to brackish or marine water for Vibrio spp., disrupt the tight junctions of the intestinal cells, effectively freshwater for Aeromonas spp. decreasing tissue resistance. Now, the action of this soil or water for C. violaceum cholera toxin is that it will trigger the hypersecretion of Transmission to humans is by ingestion of water and electrolytes by mucosal cells, leading to contaminated water, fresh produce, meat, dairy production of a profuse watery diarrhea, termed as rice products, or seafood or by exposure of disrupted skin water stools or rice watery stools, that are composed of and mucosal surfaces to contaminated water. fluids and mucous flecks and is the hallmark of cholera VIBRIO CHOLERAE toxin activity. causes epidemics and pandemics Now, production of this watery diarrhea can produce a diarrheal disease cholera dramatic fluid loss to the patient, leading to severe Reservoir: humans dehydration and hypotension. And if these fluids are not replaced, that can lead to death. Has dormant stages Vibrio cholerae, which is the agent of a diarrheal disease Somatic antigens known as cholera, or known as Asiatic cholera, or ü V. cholerae 01, epidemic cholera. Since 1817, the world has witnessed ü V. cholerae 0139 seven cholera pandemics. ü V. cholerae non-01/non-0139 In 2010, after a devastating earthquake in Haiti, more than The somatic antigens O1 and O139 associated with the 600,000 infections and 7,000. deaths occurred as a result Vibrio cholera cell envelope are positive markers for of Vibrio cholerae O1 infections within a 24-month period. strains capable of epidemic and pandemic spread of the During these outbreaks, the organism was spread among disease. Strains carrying these markers almost always people by the fecal-oral route due to poor sanitation. produce cholera toxin, whereas non-O1 or non-O139 Evidence suggests that the Bacillus has a dormant stage strains do not produce the toxin and hence do not produce that allows its long-term survival in brackish water or cholera. The non-O1 and non-O139 strains are saltwater environments during interacademic periods. associated with non-epidemic diarrhea and extraintestinal These dormant stages are considered viable but not infections. culturable. Vibrio parahaemolyticus Virulence factors second most common Vibrio species implicated in ü Cholera toxin (CT) / choleragen gastroenteritis ü zonula occludens (Zot) toxin (enterotoxin) Summer diarrhea in Japan ü accessory cholera enterotoxin (Ace) toxin serotype 03: K6 - pandemic strain ü 01 and 0139 somatic antigens consumption of raw, improperly cooked, or ü hemolysin/cytotoxins decontaminated seafood, particularly oysters. ü motility, chemotaxis, mucinase, and toxin Kanagawa phenomenon coregulated pili (TCP) pili PANANGGILAAN, J. It has also been observed that most clinical Vibrio para GRAM STAIN hemolyticus strains produce a heat-stable hemolysin or kanagawa that is able to lyse human erythrocytes in a gram-negative, straight or slightly curved rods high-special, high-salt mannitol medium we call as rapid darting or shooting-star motility (dark field wagatsuma agar. Now these strains are considered microscopy) kanagawa toxin positive, whereas most environmental CULTURE MEDIA isolates are kanagawa toxin negative. thiosulfate citrate bile salts sucrose (TCBS) agar - Vibrio vulnificus stool cultures Second most serious types of Vibrio-associated ü Sucrose fermenters - yellow colonies infections ü Non - sucrose fermenters - green Two categories, primary septicemia and wound Alkaline peptone water (pH 8.4) - enrichment broth infections MAC / SS: non-lactose fermenters except V.vulnificus Consumption of shellfish, especially raw oysters o Perform oxidase test from SBA plate Patients with liver dysfunction and syndromes that result For culture media, we have your thiosulfate citrate basalt in increased serum level of iron, such as in sucrose agar, which contains 1% sodium chloride, bile hemochromatosis, cirrhosis, thalassemia major, and salts that inhibit the growth of gram-positive organisms, hepatitis are particularly predisposed to this scenario. and sucrose for the differentiation of the various Vibrio Patients with Vibrio vulnificus wound infections invariably species. have a history of some type of traumatic aquatic wound Bromothymol blue and thymol blue pH indicators are that often presents as cellulitis. added to the medium, and the high pH of the medium, Vibrio alginolyticus which is 8.6, inhibits the growth of other intestinal microbiota. So, in your TCBS, the sucrose fermenters least pathogenic for humans present as yellow colonies, while the non-sucrose originate from extraintestinal sources fermenters present as green colonies. Alkaline peptone occupational hazard for people in constant contact water with a pH of 8.4 may be used as an enrichment with seawater, such as fishermen or sailors broth for obtaining growth of Vibrios from stool. Specimen Collection and Transport After inoculation, the broth is incubated for 5 to 8 hours at 35 degrees Celsius, and then subcultured to TCBS. Now, whenever possible, body fluids, pus, or tissues Oxidase testing is unreliable when performed on colonies should be submitted. But swabs are acceptable if they are grown on TCBS medium. Now, for your MAC and transported in an appropriate holding medium, such as: Salmonella shigella agar, they also may grow on this Ö Cary-blair – prevent desiccation media and present as non-lactose fermenters, except for Vibrio vulnificus. X Buffered glycerol saline Now, in case if we are evaluating an SBA plate or a sheep Strips of blotting paper soaked in liquid stool and placed blood agar plate, you must perform an oxidase test, in airtight plastic bags with a few drops of saline to because lactose-positive colonies from selective maintain moisture are considered viable specimens for up differential media such as MAC agar can give false- to 5 weeks. Buffered glycerol saline is not recommended positive oxidase reaction. The yellow colonies that as a transport or holding medium because glycerol is toxic ferment sucrose is your Vibrio cholerae, and the green for Vibrios. ones are your Vibrio parahemolyticus. DIRECT DETECTION METHODS ELISA for V. cholerae toxin available latex agglutination test Rapid antigen tests and immunochromatographic Assays are available PANANGGILAAN, J. STRING TEST are halophilic or salt-loving, and require the addition of sodium for growth. 0.5% sodium deoxycholate lyses Vibrio cells, but not those of Aeromonas spp. Okay, so this is where the, there's no string test here, but as I mentioned, it will differentiate Vibrio from your Aeromonas, There is cell lysis that will release your DNA, which can whereas it's also to differentiate cholerae, the non-O1 and O1- be pulled up onto a string with an inoculating loop. So, 2-9, then O1 and the O1-2-9, non-O1-2-9 strains could be your Vibrio will lyse, the Vibrio cells will be lysed, but not through your Vibrio static test. Okay, and interspecies of Aeromonas spp. So, Vibrio will display a positive string differentiation can be done through growth in sodium chloride, and production of acid from lactose, sucrose, and arabinose. test, whereas Aeromonas will give a negative result. AEROMONAS VIBROSTATIC TEST (0129) oxidase-positive, glucose fermenting, gram-negative Vibrios - Susceptible rods Oxidase + glucose fermenters - Resistant widely distributed in freshwater, estuarine, and V.cholerae 01 - Susceptible marine environments worldwide. V.cholerae non-01 – Resistant isolated from retail produce sources and animal meat products. typically grow from 10° to 42° C. classified into one of two groups ü mesophilic group - A. hydrophila complex, A. veronii complex, A.caviae complex ü psychophilic group - A. salmonicida We have the mesophilic group, which has an optimal growth of around 37 degrees Celsius. These organisms are considered mesophiles because they grow well at 37 degrees Celsius. They are all motile by means of a single polar flagellum, and under this group, we have the Aeromonas hydrophila complex, the Aeromonas veroni complex, and the Aeromonas caviae complex. Under the psychrophilic group, which grows optimally around 22 degrees Celsius, we have your Aeromonas salmonicida, which is a fish pathogen with several subspecies. This organism is non-motile and grows best at 22 to 25 degrees Celsius. Psychrophilic non-motile strains are not considered human Vibrios can be easily confused with other genera, pathogens. including Aeromonas, Plesiomonas, Pseudomonas, and CLINICAL MANIFESTATIONS the members of the family Enterobacteriaceae. Their general susceptibility to the Vibriostatic agent O1-2-9, not Intestinal Infections in this table, it's not found in this table, but it's in another table, and positive string test, also not in this table, ü A.caviae - neonate and pediatric populations. distinguishes them from Aeromonas. Their inability to ü A.hydrophila - HUS or kidney disease ferment inositol, except from your Vibrio cincitnatiensis, ü A.veronii biovar sobria - also HUS and cholera-like disease and some strain of your Vibrio micinicovi, separates them from your Plesiomonas. Extraintestinal Infections Their positive oxidase reaction, except for Vibrio ü septicemia and wound infections are the most micinicovi again, separates them from the common - A. hydrophila subsp. Hydrophilia Enterobacteriaceae, excluding your Plesiomonas ü surgical wound infections involving the use of leeches sigilloides. And carbohydrate fermentation metabolism for medicinal therapy after plastic surgery to relieve separates them from the oxidative Pseudomonas. All venous congestion - A. veronii biovar sobria species, except for Vibrio cholerae and Vibrio mimicus, PANANGGILAAN, J. Usually, these are cholera-like disease. And as for extra- distinguishes Aeromonads from oxidase-positive non- intestinal infections, we have septicemia and wound fermenting pseudomonas isolates. infections are the most common by your Aeromonas hydrophila, subspecies hydrophila. Now, wound infection invariably involves a recent traumatic aquatic exposure, such as boating or fishing accident, and generally occurs on the extremities. This is the most common presentation is cellulitis. Now, Aeromonas sepsis is one of the most invasive type of Aeromonas infection and similarly has a strong association with the species A. veroni biobarsubria, A. jandei, and A. hydrophila. Such patients are most likely to be immunocompromised. Aeromonas veronis biovar subria is also implicated in surgical wound infections involving the use of leeches for medicinal therapy after plastic surgery to relieve venous congestion CULTURE MEDIA A.hydrophila, A. veronii biovar sobria, and A. jandaei, display strong B-hemolysis most commonly isolated species, A. caviae Ferments lactose in MAC CAMPYLOBACTER May resemble Yersinia enterocolitica in CIN Microaerophilic PRESUMPTIVE IDENTIFICATION Oxidase positive (concentration less than that of Oxidase positive room air. 5% is optimal) Indole - positive Gram negative curved shaped bacilli grow well in nutrient broth with 0% NaCl, but not in Assacharolytic 6% NaCl Campylobacter and Campylobacter-like species, which Ferments glucose with or without gas include Calicobacter and Wolinella, have undergone So, perform an oxidase test, especially taken from your changes in taxonomy. Campylobacters were formerly sheep blood agar, to easily separate the oxidase-positive classified with the Vibrios because of their positive Aeromonas from the oxidase-negative Yersinia, oxidase and characteristic microscopic morphology. But especially also on CIN because they may resemble your DNA homology studies showed that Campylobacter Yersinia enterocolitica. Most Vibrios, specifically the species do not belong with the Vibrios. In addition, unlike halophilic species, cannot grow in 0% NaCl but thrive in the Vibrios, which are fermentative, most Campylobacters 6% NaCl and even higher concentration of NaCl. are asaccharolytic, meaning they do not utilize any sugar. However, because your Vibrio cholerae and Vibrio HELICOBACTER PYLORI mimicos are non-halophilic and grow well without additional salt, any salt-tolerance test must be used in strongly associated with gastric, peptic, and duodenal conjunction with the string test and O129 disc to ulcers as well as with gastrointestinal carcinoma. distinguish Aeromonads from this major pandemic High incidence cholera species and the less common sucrose-negative type B gastritis Vibrio mimicos. H. cinaedi Fermentation of inositol can be used for separating Aeromonads from pliceomonads, in which Aeromonads are negative and pliceomonads are positive. Finally, the ability to ferment glucose with or without production of gas PANANGGILAAN, J. EPIDEMIOLOGY cause abortion in domestic animals, such as cattle, sheep, and swine and are primarily zoonotic organisms most common cause of bacterial gastroenteritis worldwide - Campylobacter jejuni. C. fetus - blood cultures Guillain-Barré syndrome It is estimated that 50% of the population worldwide is Select there are four selective media for the cultivation of infected with Helicobacter pylori. In developing countries Campylobacter species, each with a specific base and a in Africa, Asia, and South America, the incidence is certain antimicrobial agent. So we have Campy blood reported to be as high as 80% to 90%. This higher agar plates, Keros, Butzler, and CCDA. So Campy blood incidence is attributed to poor sanitary conditions, and it agar or Campy blood agar is a commonly used medium appears that infection occurs early in life. to isolate Campylobacter gengione and other enteric Campylobacters. It's also the major cause of type B gastritis, which is a chronic condition formerly associated primarily with stress This commercially available medium contains Brusella and chemical irritants. And Helicobacter cinaedi has been agar as base, 10% sheep red blood cells, and a isolated from the blood of patients with bacteremia and combination of antimicrobials which are your patients with HIV infection. Vancomycin, Trimetoprim, Polymyxin B, and Ampotericin B and Cipalotin. Other selected media that have been Campylobacter jejunii is considered the most common cause of bacterial gastroenteritis worldwide. The successful in recovering Campylobacter, as I said, are transmission of your Campylobacteriosis has been your Butzler medium and your Skiros. attributed to direct contact with animals and handling Your Campy CBA or your Cephoperazone-Vancomycin- infected pets such as dogs, cats, and birds and indirectly Ampotericin Bmedium has been reported to provide better by the consumption of contaminated water and dairy suppression of your fecal biota even when this medium is products and improperly cooked poultry. Person-to- incubated at 37 degrees Celsius. Incubation at 37 person transmission has been reported and to some degrees Celsius allows the recovery of Campylobacter Campylobacter species are also sexually transmitted. species that are inhibited at 42 degrees Celsius. There is strong evidence suggests that Campylobacter Campylobacter fetus, subspecies fetus, Campylobacter infection plays a role in Guillain-Barre syndrome, which is rectus, and Campylobacter curvus can be isolated using an autoimmune disorder characterized by acute paralysis routine culture media. Your charcoal-based blood-free caused by damage to the peripheral nervous system. media such as your charcoal Cephoperazone Many patients with GBS test positive for antibodies to deoxycholate agar or CCDA and the charcoal-based Campylobacter. selective media, CSM, are also available. SPECIMEN COLLECTION AND TRANSPORT A combination of media that contain either CCDA or CSM can achieve the highest yield of Campylobacter species Blood culture - C. fetus subsp. fetus in stool samples. To recover your Campylobacter pylori, a Stool samples combination of a non-selective medium such as your Cary-blair chocolate agar or Bruxella agar with 5% horse red blood Avoid buffered glycerol saline cells and a selective medium such as Skiros agar may be H. pylori - gastric biopsy materials used. Stuart medium Tissue samples may also be placed in cysteine- It is important that the inoculated medium be fresh and Brucella broth with 20% glycerol and frozen at - 70° C moist and that the culture be incubated in a microaerophilic environment with increased humidity. Again, microphilic environment only requires 5% oxygen. PANANGGILAAN, J. INCUBATION A positive heparate hydrolysis test is an important characteristic for the identification of Campylobacter Incubate stool cultures at 42° C to recover C. jejuni. jejuni. Now interspecies identification, aside from growing Inhibitory for C. fetus subsp. Fetus - grow instead at at various temperature changes such as temperature 37° C range, such as 25 degrees Celsius, 37 degrees Celsius, Enteric Campylobacter and Helicobacter spp. require and 42 degrees Celsius, each species can also be a microaerophilic and capnophilic environment. differentiated whether they are susceptible or resistant to As for incubation, since we already emphasized that it Nalidixic acid and Cephalothin. must be a microphilic environment, it must also be DEFINITIVE IDENTIFICATION capnophilic. So that's the reason why the ideal atmospheric environment for this organism is a gas mixture of 5% oxygen, 10% carbon dioxide, and 85% nitrogen gas for Campylobacter. For your Helicobacter, it's 5% to 10% oxygen, 5% to 12% carbon dioxide, and 5% to 12% carbon dioxide. As for the stool cultures, incubate at 42 degrees Celsius to increase the recovery of your Campylobacter jejuni. However, this temperature is inhibitory for your Campylobacter fetus H. pylori can be presumptively subspecies fetus. identified in a gastric biopsy specimen by testing for the presence So it's recommended that if you're suspecting of a rapid urease reaction Campylobacter fetus, grow it instead at 37 degrees Urea breath test Celsius. Now on gram stain, this organism will stain poorly. So for better visualization, carbolfoxin is recommended as a counterstain instead of your safranin. Definitive identification of Helicobacter pylori can be done If you're going to be using safranin still, because through testing for rapid urease reaction on a gastric carbolfoxin is not available, counterstain it should be biopsy. So here, the collected tissue sample is placed extended to 2 to 3 minutes. It will be longer than the usual onto a Christensen's urea medium and is incubated at 37 gram staining. degrees Celsius for 2 hours. A color change suggests the MICROSCOPIC MORPHOLOGY presence of Helicobacter pylori. Now, urease activity can also be detected by the urea curved, non-spore-forming, gram-negative rods breath test, which is reportedly sensitive and specific and long spirals or S- or seagull-wing shapes is recommended for monitoring therapy. In this test, the Corkscrew motility patient is given 39 carbon or 49 carbon labeled urea orally. Urea is then degraded by the urease activity of Helicobacter pylori in the stomach and releases as 13 carbon dioxide or 14 carbon dioxide, which is absorbed into the bloodstream and is detected in the exhaled breath Isolates from stool specimens and rectal swabs can be by a scintillation counter. presumptively identified as Campylobacter species by a positive oxidase, the characteristic gram stain IMMUNOLOGIC ASSAYS microscopic morphology, and the characteristic motility. The microscopic morphology is important because it Latex agglutination tests are available but cannot differentiates Campylobacter from other bacteria such as differentiates among the Campylobacter spp. Aeromonas and Pseudomonas, which are oxidase ELISA / IIFA positive and can grow at 42 degrees Celsius in a Now, there are also immunologic assays available for microaerophilic environment. your Campylobacter and Helicobacter, such as latex agglutination test, but it cannot differentiate among the PANANGGILAAN, J. Campylobacter species, so still, we employ the manual method of culture. And also, specific antibodies in serum can be detected by enzyme-linked immunosorbent assay or indirect immunofluorescent assay methods. These methods have been reported to be reasonably sensitive and specific indicators of Campylobacter and Helicobacter pylori infections. Serologic testing is useful for epidemiologic studies for Campylobacter but is not recommended for routine diagnosis. Serologic testing is an important screening method, though, for the diagnosis of Helicobacter pylori infection. >>>>>>>>>>>>>>>>>>>>END>>>>>>>>>>>>>>>>>>> PANANGGILAAN, J.