and . MacConkey Medium. However, cases of human-to-human transmission have been reported. The epidemiology of the infection and the di The addition of salt supplies the important electrolytes needed for transport and osmotic balance. Nei casi rari in cui infetti luomo, la maggior parte dei ceppi di P.fluorescens sono sensibili alla tetraciclina, agli aminoglucosidi e alle nuove beta-lattamine, ureidopenicilline, carbapenemici, monobattami. 2000). Main diseases caused by the major pathogenic Pseudomonas, Burkholderia and Stenotrophomonas species in veterinary medicine Functions Kocuria rhizophila- isolation from a free weight at UAF and These shapes include spherical, rod . The red color is due to production ofacid from lactose, absorption of neutral red and a subsequent color change of the dye when the pH of medium falls below6.8. This pathogen is a facultative intracellular bacterium which has a wide host range, including humans, horses, sheep, goats, dogs, cats, cattle, and pigs. Il nome P. A pure bacterial culture remains crucialto assess its virulence, its susceptibility to antibiotic therapy, and its genome sequence in order to facilitate the understanding and treatment of caused diseases. mycoplasma PPLO, Your example of vulgaris and aureus look like they have been switched to me. Genus Characteristics As pyocyanin is unique to P. aeruginosa this is an important diagnostic characteristic although strains vary in the amount of the pigment they produce. Some strains produce all four pigments. Thank you sir for your excellent notes, it really guides me in the treatment plant where im working as a microbiologist in the lab. P. fluorescens was recovered from port catheters in 17 (81%) of 21 patients, from blood cultures in 15 (79%) of 19 patients, and from both blood and catheter cultures in four (33%) of 12 patients. Il MacConkey contiene inoltre come unica fonte energetica illattosio: ci permette quindi la distinzione tra batteri fermentanti il lattosio e batteri che non fermentano questo zucchero. Colonial morphology PPT - Pseudomonas PowerPoint Presentation, free download - ID:2269529 It is easily killed by desiccation, sunlight and common disinfectants. putida. Agar 13.5 g/L pH finale 7.10.2 a 25C Preparazione del terreno - disciogliere 50g di terreno in polvere in un litro d'acqua distillata fredda; - portare ad ebollizione sotto agitazione e quindi mettere in autoclave a 121C per 15 minuti; - estrarre, lasciar raffreddare (fino a circa 50C) e quindi versare in piastre sterili. do not fluorescens under UV light and grow poorly [7]. The respiratory tract is the most common site of infection. Human disease cases can present as localized, suppurative cutaneous infections, pulmonary infections, bloodstream infections or suppurative chronic infections of the skin. Dopo aver piastrato la sospensione batterica, la piastra viene incubata a 25C per 48 ore. Biology 2EE3 - Lab 1 Winter 2023 1.2 Growth of bacteria on MacConkey agar In this lab, you will grow four bacterial cultures (Lactose-positive Escherichia coli, Lactose-negative Escherichia coli, Micrococcus luteus, and Pseudomonas fluorescens) on MacConkey agar to assess some of their physiological and metabolic properties. Incubation of MacConkey Agar plates under increased CO2has been reported to reduce growth andrecovery of a number of strains of Gram-negative bacilli. MacConkey Agar Is Selective for Non-fastidious Gram-negative Organisms Therefore, MacConkey needed a way to limit this background of environmental flora and allow only his organisms of interest to grow. I batteri che fermentano il lattosio si presentano sotto forma di colonie con varie sfumature di rosso poich producono acidi misti che fanno scendere il pH. FIELD: biotechnology.SUBSTANCE: nutritive medium for cultivation Pseudomonas fluorescens AP-33 contains molasses, potassium phosphate disubstituted three-water, magnesium sulphate seven-water, polished peas, pre-treated by autoclaving, succinic acid, Polypropylene glycol for a liquid medium, and distilled water at specified ratios of components . 1972; Palleroni 1993; Kersters et al. Add 5ml of glycerol. Certolizumab Pegol e il suo ruolo contro il TNFa, Obesit in et pediatrica: una patologia da non sottovalutare, 28 febbraio 1953: 70 anni fa la scoperta del DNA. Sterilize by autoclaving at 15 lbs pressure (121C) for 15 minutes i.e. Analista presso impianto di potabilizzazione dell'acqua. The genome contains numerous insertion sequence elements and a vast number of simple sequence repeats. Cool to 45-50C. Transmission occurs from infected animals via contaminated food and water and less commonly from aerosols and infection of wounds. You may also needAeromonas, Plesiomonas and Vibrio speciesBacillus speciesErysipelothrix speciesMiscellaneous Gram-negative bacteriaEnterobacteriaceaeMycobacterium speciesLawsonia intracellularisCampylobacter, Arcobacter and Helicobacter species Pancreatic digest of gelatin and peptones (meat and casein) provide the essential nutrients, vitamins and nitrogenous factorsrequired for growth of microorganisms. Morphology & Culture Characteristics of Pseudomonas aeruginosa 1994). ( 3) per essere sottoposte alle prove biochimiche quali il test dellossidasi, il test di fermentazione/ossidazione degli zuccheri. Infected Equidae are the reservoir for B. mallei. MacConkey agar (MAC) was the first solid differential media to be formulatedwhichwas developed at 20th century by Alfred Theodore MacConkey. As P. aeruginosa is resistant to many antimicrobials, it frequently causes infection in animals undergoing antibiotic treatment or in immunocompromised hosts. Si aggiunge cfc supplement per rendere il terreno maggiormente selettivo.LAcido Fusidico inibisce i Gram-positivi e la Cefaloridina un antibiotico a largo spettro. In intensive care units, testing multi-drug resistance in gram-negative bacteria is an important surveillance measure. Migula. In humans, the clinical signs of melioidosis (also referred to as Whitmores disease) vary greatly from an asymptomatic presentation to a fatal septicaemia. Pseudomonas testosteroni (now Comamonas testosteroni ) MICROSCOPIC APPEARANCE MACROSCOPIC APPEARANCE Some colonies produce water-soluble pigment pyoverdin or pyocyanin that fluoresce white to beige under UV light. Pseudomonas Media and Tests - Sigma-Aldrich These may also be atypical in certain biochemical reactions, making them difficult to identify. Per lisolamento di P.fluorescens da campioni alimentari contaminati si utilizza il terreno selettivo Pseudomonas Agar Base (PAB) addizionato con cetramide, fucidina e cefaloridina (CFC supplement). Pseudomonas gives negative Voges Proskauer, indole and methyl red tests, but a positive catalase test. Transmission can also occur via arthropod bites. ), which permits others to distribute the work, provided that the article is not altered or used commercially. qual la metodica analitica di riferimento? Se presenti le colonie si presentano molto piccole e opache. The disease usually occurs in tropical regions between 20 northern and southern latitudes but melioidosis has been reported in localized areas of France, Iran, China and the USA. Preparation of MacConkey Agar Weigh and suspend 50 grams of MacConkey agar powder in 1 Litre of purified water and mix thoroughly. This gives McConkey agar its differentiating property. Pseudomonas aeruginosa is an opportunistic pathogen that infects burns, wounds, surgical incisions and sites of catheterization. Many Gram-negative pathogens regulate virulence factor expression by using a cell density mechanism termed quorum sensing. Human infection has occurred rarely and sporadically among laboratory workers and also those in direct contact with infected, domestic animals. 2006). 2004). Humphries RM, Linscott AJ. The genus Pseudomonas (sensu stricto) represents the rRNA homology group 1 with the type species Pseudomonas aeruginosa which is the most important veterinary pathogen in the genus Pseudomonas followed by P. fluorescens. Red colonies and medium, indicative of an alkaline reaction, are seen on brilliant green (Fig. Sodium chloride maintains the osmotic balance in the medium. Altogether, MacConkey agar only grows gram-negative bacteria, and those bacteria will appear differently based on their lactose fermenting ability as well as the rate of fermentation and the presence of a capsule or not. Directions. However, direct microscopy of Gram-stained smears with B. pseudomallei will often reveal small Gram-negative bacilli with bipolar staining, safety pin appearance. Pseudomonas and Stenotrophomonas can be handled in a biosafety level-2 laboratory. 2001). This pathogen is a facultative intracellular bacterium which has a wide host range, including humans, horses, sheep, goats, dogs, cats, cattle, and pigs. Initially couldnt understand this MacConkey that i encountered in the microbiology lab but now i can understand its uses. MacConkey agar with bile salt- Selective for gram-negative bacteria but . P. fluorescens, P. putida, P. luteola). What color is Pseudomonas aeruginosa on MacConkey agar? It is nonsporeforming and is motile by one or more flagella. no. Pyorubin and pyomelanin are less commonly produced, develop slowly and are seen best by growing the strains on nutrient agar slants at room temperature for up to two weeks. Pseudomonas fluorescens Heavy growth may produce a greenish-gold fluorescent pigment. Therefore, lactose-fermenting-gram-negatives (lactose-fermenters) will form pink colonies, while non-lactose fermenters will form off-white opaque colonies. Update: Delayed Onset <I>Pseudomonas fluorescens </I>Bloodstream (1, 6, and 10) Burkholderia pseudomallei is the aetiological agent of melioidosis, a disease in which treatment failures and relapses are common, with pneumonia as the most common clinical presentation. putida (6). The toxins are involved in epithelial cell damage and in the inhibition of phagocytic cells. The entry, colonization, and infection by P. aeruginosa depend on a number of factors; termed virulence factors that assist the survival of the organisms and evasion of host defense. P. fluorescens conosciuto principalmente per aver creato problemi nellindustria alimentare, presenta diversi fattori di virulenza che rendono i ceppi anche patogeni per luomo. Tatuaggio infetto: come riconoscerlo e cosa fare? Pseudomonas fluorescens is an unusual cause of disease in humans, and usually affects patients with compromised immune systems (e.g., patients on cancer treatment). Inoculate stool in selenite F broth or tetrathionate broth and incubate. Interestingly, pyocyanin can colour pus and stain wool a greenish blue. Biologa. Flagellum In questo terreno, infatti, sono presenti sia il cristalvioletto che i sali biliari i quali sono in grado di inibire la crescita dei batteri Gram positivi (ed anche quella dei Gram negativi pi esigenti dal punto di vista nutrizionale). Cytotoxic, invasion of tissue and cellular damage, immunosuppressive action Grazie per linteresse! The skin form is also characterized by the formation of tubercle-like nodules, usually along the cutaneous lymphatics, which frequently ulcerate. 18.4) in varying combinations and amounts. Burkholderia pseudomallei is the causative agent of melioidosis or pseudoglanders. Related Burkholderia mallei, the aetiologic agent of glanders, is a listed disease by the Office International des pizooties (OIE), also known as the World Organization for Animal Health. Lastly, some species that forms a capsule appear differently. The typical lesions are nodules which may suppurate and can form in any tissue, including the brain. Pseudomonas can grow at wide ranges of temperature; the optimum temperature is 37 C. It can grow on ordinary media like nutrient agar and grows almost on all the culture media used routinely in the bacteriology lab. To 500ml of agar base cooled to 50C add the contents of 1 vial of Pseudomonas CN Supplement (SR0102) rehydrated as directed. The growth of B. mallei is enhanced by 1% glycerol. 2003, Althouse & Lu 2005). Exotoxin A (ADP-ribosyl-transferase) Bi ging Vi sinh vt i cng (Phn 2) why do staphylococcus aureus look dark red and smooth on Mac conkey agar. Just curious. The manifestations of the disease depend on the extent and distribution of the lesions in the animal. Some strains have colonies with a distinctive metallic sheen (Fig. In animals, infections are usually systemic and chronic but acute disease with terminal septicaemia may occur. MAC contains a pH indicator that turns pink under acidic conditions. Ps. CFC Selective Supplement is an antibiotic supplement for the selective isolation of . If MAC is for non-fastidious Gram-negative bacilli, why would Enterococcus and Staphylococcus give a reaction on this medium? Crystal violet dye and bile salts halt the growth of gram-positive bacteria. The culture was isolated from pre-filter tanks and has applications in bioinformatics, water testing, and antimicrobial preservative testing. Consequently, DNA/RNA hybridization confirmed the presence of five diverse rRNA groups (rRNA groups I-V) (Palleroni et al. The Origin of MacConkey Agar - ASM.org Burkholderia mallei once had a wide geographical distribution but now is mainly seen in China and Mongolia with pockets of infection in India, Iraq, Turkey and the Philippines. Most strains give a clear zone of haemolysis on blood agar (Fig. Please help me, pasteurella multocida (gram negative coccobacilli) does not grow on MacConkey .please explain, Good evening sir I dont understand you said macconkey agar inhibits the growth of gram positive bacteria but you are saying that staphylococcus gives a pale pink colour on this culture midum. Buongiorno, Mix well before pouring into sterile Petri plates. Burkholderia mallei once had a wide geographical distribution but now is mainly seen in China and Mongolia with pockets of infection in India, Iraq, Turkey and the Philippines. Primary lesions occur at the point of entry (skin or mucosal surfaces) with dissemination via the lymphatic system and dissemination by the bloodstream. Abbott IJ, Jenney AW, Spelman DW, Pilcher DV, Sidjabat HE, Richardson LJ, Paterson DL, Peleg AY. Jung B, Hoilat GJ. Access free multiple choice questions on this topic. Example of mucoid colony-forming species: Elazhary MA, Saheb SA, Roy RS, Lagac A. Figure 18.1 Pseudomonas aeruginosa on sheep blood agar showing large, flat, irregular-edged colonies resembling those of some Bacillus species. I salute you for the nice explanations in Microbiology you regularly upload in Facebook. A major function of this secretion system is to secrete virulence-associated proteins into target cells of the host organism. Burkholderia mallei, the aetiologic agent of glanders, is a listed disease by the Office International des pizooties (OIE), also known as the World Organization for Animal Health. Motility, adherence to mucin Clinical microbiologists must be trained with a thorough understanding of the principles to run the tests, interpret the results, and report the findings. When bacterial gastroenteritis is suspected, the patient's specimen can be sampled and cultured on a panel of bacterial cultures, which includes MacConkey medium. 1989; Swain etal. The colonies of P. aeruginosa are large (34mm), flat, spreading, greenish-blue with a serrated edge and a characteristic fruity, grape-like odour of aminoacetophenone. grazie mille. However, cases of human-to-human transmission have been reported. Studies have indicated that virulence of selected B. pseudomallei isolates is variable and dependent on factors such as iron bioavailability, inoculum size and host risk factors (Ulett etal. The toxins are involved in epithelial cell damage and in the inhibition of phagocytic cells. MacConkey agar is used for the isolation of gram-negative enteric bacteria and the differentiation of lactose fermenting from lactose non-fermenting gram-negative bacteria. Pseudomonas are aerobic, rod-shaped, motile, non-spore-forming flagellated (one or more polar flagella), gram-negative bacteria. PDF Lesson-28 - National Institute of Open Schooling Mix well and pour into sterile Petri . Agar MacConkey - Wikipedia Why does pseudomonas aeruginosa grow on MacConkey agar despite - Study 2007). These bacteria are environmental microorganisms typically found in water, soil, on plants, fruits and vegetables. Figura 2 La filogenesi di Pseudomonas fluorescens. The colonies on the cetrimide agar appear dry and flat with a circular circumference and lobular edges. Pseudomonas aeruginosa - Wikipedia Category:Pseudomonas fluorescens - Wikimedia Commons MacConkey agar is designed to grow Gram-negative bacteria and also contains crystal violet dye which inhibits the growth of Gram-positive bacteria. Media such as Pseudomonas agar P (BD Diagnostics) (Fig. Some strains may be encountered that grow poorly or fail to grow on this medium. Pseudomonas, Stenotrophomonas and Burkholderia species have a worldwide distribution. The section in this information sheet entitled "Preventive actions and reducing risk" communicates updates in risk . Members of the Pseudomonas, Burkholderia and Stenotrophomonas genera grow in broth blood culture systems within the five-day standard incubation guideline. Burkholderia mallei is a host-adapted pathogen, causing glanders (pulmonary and nasal forms) or farcy (the skin form) in the Equidae population. MacConkey Agar: Introduction, Principle, Composition, Preparation Biochemical Test and Identification of Pseudomonas aeruginosa Human infection has occurred rarely and sporadically among laboratory workers and also those in direct contact with infected, domestic animals. These virulence factors all play a role in disease pathogenesis (Table 18.2). Pseudomonas fluorescens strain NCTC 10038 is a whole-genome sequenced bacterial type strain. Agar isthe solidifying agent. Some species produce soluble pigments and most will grow on MacConkey agar as lactose non-fermenters as well as converting nitrate to nitrite or nitrogen gas. Flagella provide motility and act as adhesins to mucin and respiratory epithelial cells. Siderophores are involved in iron acquisition and promote survival in low-iron conditions such as host tissues. It has emerged as an opportunistic pathogen in animals and immunocompromised humans. 2001). grows only in the presence of oxygen.. There are various culture media used for the cultivation of Pseudomonas aeruginosa (P. aeruginosa) in the laboratory and most commonly the Nutrient Agar medium and MacConkey Agar medium is used, the other media are as follows - This bacterium is frequently found in aerators and traps of sinks. Pseudomonas aeruginosa produces large, pale colonies on MacConkey agar (unable to utilize lactose) with greenish-blue pigment superimposed (Fig. Escherichia coli and Campylobacter jejuni are some of the common causes of bacterial gastroenteritis. Assistant Professor of Agricultural Microbiology, are thin, rod-shaped, non spore-forming gram-negative bacilli with a guanine/cytosine content of 57-68 mol%. Gram-positive bacteria will not form any colonies on MacConkey medium. To Prepare Pseudomonas CN Agar. Protection from phagocytosis, adhesin, antimicrobial resistance Burkholderia species will also grow on MacConkey agar, with the exception of B. mallei. The growth of B. mallei is enhanced by 1% glycerol. The colonies of P. aeruginosa are large (34mm), flat, spreading, greenish-blue with a serrated edge and a characteristic fruity, grape-like odour of aminoacetophenone. Under particular conditions, P. aeruginosa can produce an alginate structure which is a slime-like, mucoid exopolysaccharide. Le colonie si presentavano lisce, traslucide e biancastre; sul terreno PDA si poteva riconoscere 2 colorazioni differenti: un pigmento blu intenso e fluorescente legato alla cellula e un pigmento idrosolubile verdastro diffuso nel terreno. The degree of. Colony morphology of Pseudomonas fluorescens, after 24 hrs growth on Neutral red is a pH indicator that turns from off-white to bright red/pink as the pH drops below 6.8. Based on the ability to ferment lactose, different species will yield colonies in varying appearance on a MacConkey medium. The Final Screening Assessment for the Pseudomonas fluorescens strain ATCC 13525 was published in February 2015 under the Chemicals Management Plan (CMP). StatPearls Publishing, Treasure Island (FL). Colonial variation includes smooth, soft and shiny (S-forms), dwarf, dry and granular (R-forms) not unlike some colonies of Bacillus species, and mucoid (M-forms) that are frequently biochemically atypical. Its name come from the production of the soluble fluorescent pigment pyoverdin. 28.2: Pseudomonas aeruginosa on nutrient agar showing greenish . Malleobactin is a siderophore involved in iron acquisition (Alice etal. Check turbidity and subculture on solid media like blood agar and MacConkey agar. However, selective media which inhibit the growth of P. aeruginosa is recommanded for the recovery of B. pseudomallei (Ashdown agar or broth with colistin) and B. cepacia (PC, OFPBL, and BCSA agars). Weak lactose fermenters will form colonies slower than the rest. Strains of P. aeruginosa produce the water-soluble diffusible pigments pyocyanin (blue, phenazine pigment), pyoverdin (water-soluble yellow-green or yellow-brown pigment), pyorubin (red) and pyomelanin (dark brown) (Fig. fluorescens. Some strains of P. aeruginosa do not produce pigments and are highly mucoid. Predisposing causes include trauma to tissue (burns and wounds), debilitation due to malignancy or immunodeficiency and an imbalance in the normal flora, often caused by antibiotic therapy. 70143, because . Its capsular polysaccharide is reported as a major virulence factor (DeShazer etal. Bacillus subtilis No or very little growth. While working for the 'Royal Commission' on sewage disposal, his role was to inspect drinking water sources for gram negative enteric organisms. Pseudomonas has been seen to grow in distilled water, also. Pseudomonas aeruginosa is a common encapsulated, gram-negative, aerobic-facultatively anaerobic, rod-shaped bacterium that can cause disease in plants and animals, including humans. A simple screening technique for the detection of Pseudomonas aeruginosa colonies by their fluorescence on MacConkey agar under ultraviolet light is proposed. Ci che d selettivit a questo terreno sono il cristalvioletto e i sali biliari, che inibiscono la crescita dei batteri Gram positivi. The respiratory tract is the most common site of infection. In these animals, it can be found on skin and mucous membranes, particularly the gastrointestinal tract. Download Genome Learn about our Enhanced Authentication Initiative. MacConkey agar - scheda del terreno di coltura per microbiologia I inoculated an agar slant to use as a backup culture. In humans, the clinical signs of melioidosis (also referred to as Whitmores disease) vary greatly from an asymptomatic presentation to a fatal septicaemia. You are not required to obtain permission to distribute this article, provided that you credit the author and journal. Lactose-fermenting microorganisms will produce organic acids, particularly lactic acid, which will lower the pH. are motile due to one (e.g. A species of considerable medical importance, P. aeruginosa is a multidrug resistant pathogen recognized for its ubiquity, its intrinsically advanced antibiotic resistance mechanisms, and its association with . They are common course of food contamination. Pyocyanin, a bluish pigment unique to P. aeruginosa, gives the blue-green colour associated with many cultures. Cool the medium to approximately 50C and pour into sterile Petri dishes. The type III secretion system consists of bacterial proteins which act as a syringe to deliver cytotoxins into the cytoplasm of host cells. They have a predilection for aqueous environments, surviving well in them. [10] Pseudomonas putida is a aerobic, gram negative, rod shaped bacteria. Pseudomonas fluorescens Migula - 13525 | ATCC 2004). These bacteria are strict aerobes, non-spore-forming, oxidative, catalase-positive, oxidase-positive (except P. oryzihabitans, P. luteola and the genus Stenotrophomonas) and most are motile by one or several polar flagella (except B. mallei). The use of selective media will facilitate the recovery of these bacteria from specimens with mixed flora. All bacteria were cocci-shaped except a few bacilli. Lactose fermentingstrains grow as red or pink and may be surrounded by a zone of acid precipitated bile. Oxoid - Product Detail http://creativecommons.org/licenses/by-nc-nd/4.0/ It also produces a number of extracellular products such as protein exotoxin A, proteases, type III secretion system exoenzymes, rhamnolipid, phospholipase C, and siderophores (pyochelin, pyocyanin, and pyoverdin). L'agar MacConkey contiene, inoltre, il lattosio come unica fonte di carboidrati ed il rosso neutro (un colorante che vira al rosso allorch il pH del mezzo scende al di sotto di 6,8). The green pigment pyoverdin and blue pigment pyocyanin are produced by many, but not all, strains of Pseudomonas aeruginosa. 18.5) will enhance pyocyanin production and Pseudomonas agar F (BD Diagnostics) enhances pyoverdin production (Fig. Glanders is now rare as there has been considerable success in the global eradication of this disease, principally owing to the fact that B. mallei is an obligate parasite with a restricted host range and, in addition, effective tests are available to detect carriers of the infection. It can, in rare circumstances, cause community-acquired pneumonias, as well as ventilator-associated pneumonias. I batteri che non fermentano il lattosio, invece, formano colonie trasparenti od incolori. The bacterium is capable of intracellular survival. Oxoid - Product Detail Pseudomonas aeruginosa bacteria - Microbiology in pictures Pseudomonas aeruginosa is classified as a member of the fluorescent pseudomonad group which produce pyoverdin. Some strains have colonies with a distinctive metallic sheen (Fig. http://tesi.cab.unipd.it/44793/1/Don%C3%A0_Alberto.pdf. MacConkey agar (MAC) was developed as the first solid differential media in the 20th century by a bacteriologist, Alfred Theodore MacConkey. The Pseudomonas koreensis group is classified within the Pseudomonas fluorescens complex and consist of several species, including Pseudomonas koreensis [1, 2]. From 2004 to 2006, an outbreak of P. fluorescens in the United States involved 80 patients in six states. Nei soggetti immunodepressi, P. fluorescens pu annidarsi nel tratto orofaringeo e dare infezioni post-trasfusionali e respiratorie. They grow well on standard broth and solid media such as blood agar, chocolate agar, and MacConkey agar, which are recommended to isolate Pseudomonas species from clinical specimens. Pseudomonas fluorescens - information sheet - Canada.ca Damage to tissues of the lungs and blood vessels It is selective for Gram negative species (meaning the growth of Gram positive species is inhibited). fluorescence. These will be varied and will depend on the clinical signs and lesions. 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