The pseudomonads umbrella term ¹⁶ for pseudomonas are aerobic rod-shaped, gram-negative bacteria, motile by means of one or more polar flagella. They are do not form spores. They have an absolute aerobic metabolism and the catalase positive. When aerobic bacteria such as Pseudomonas aeruginosa and facultative ‘coliforms’ grow in oxygen, glucose is completely metabolized by aerobic respiration, using it as the final electron acceptor: Glucose + 6O2→ 6H2O + 6CO2 Δ G0 = – 686 kcal/mole.¹⁵

The guanine and cytosine (G+C) content of the DNA ranges from 57 -70 mol%.B. pseudomallei (formerly Pseudomonas pseudomallei causes melioidosis (pneumoenteritis) in humans Latent infections may become active some years later due to immunosuppression.The disease is found mainly in Malaysia, Vietnam, Thailand, Myanmar, Guam, Sri Lanka, northern Australia, and the Philippines, especially in rice-growing areas and bacteria enter open wounds but can also be inhaled. B. pseudomallei is a highly infectious pathogen (Hazard Risk Group 3), therefore handle specimens with care. It is a small, motile, Gram negative rod which shows bipolar staining (like safety pins), particularly when stained with methylene blue or Giemsa stain.¹³

Gamma group – Chromatium, Francisella, Xanthomonas, Coxiella, Legionella, Pseudomonas, Acinetobacter, Moraxella, Vibrio, Aeromonas, Haemophilus, the Enterobacteriaceae + others ¹ However, in spite of the progress, the results of these studies suggested that the species assigned to the genus Pseudomonas, a number considerably smaller than years before. The number of Pseudomonas species to be included in the new edition of Bergey’s Manual of systematic bacteriology will be about 65 that the list of species should not go beyond January 2000. ⁹The strains of Pseudomonas syringae and P. fluorescens are the first genetically engineered microorganisms to be released legally in the USA were from which the ice nucleation gene was deleted to competitively exclude indigenous ice nucleation-active bacteria involved in frost damage ¹¹ .

2] Chang et al. 2001 ,high densities of Pseudomonas aeruginosa living in the insole allow for inoculation into the weight bearing area of the foot if a nail penetrates the foot through the shoe. Fisher et al. 1985; Niall et al. 1997 studied the most common atypical pathogen in this context, and a paradigm for the interaction between environmental compromise, would be the high incidence of osteochondritis to osteomyelitis, caused by Pseudomonas aeruginosa in puncture wounds of the foot associated with the wearing of training shoes (sneakers) .⁶ In 1983, Pappas et al. reported an outbreak resulting from contaminated flexible fiberoptic bronchoscopes involving 72 patients with positive cultures.. Bronchoscopes had initially been disinfected with glutaraldehyde. Matthews and Fitzsimmons 1964; Shooter et al. 1966 the problem was eventually traced to puncture of the suction channel of the bronchoscopes, resulting in contamination of the bronchoscope interior with Pseudomonas aeruginosa.⁸ Together with the genera Burkholderia and Stenotrophomonas, the species of Pseudomonas are the most opportunistic pathogens causing infections in hospitalized patients. Plasmid-determined resistance to mercuric ions (Hg2+) is very common in Pseudomonas spp found in soil and water (Bale et al. 1987). Kahyaoglu et al. 1995,a more recent potential hazard that has been highlighted is pseudomonas infection following birth in water baths & Burkholderia (Pseudomonas) cepacia, and Burkholderia pickettii, as hospital opportunists.B. cepacia grows in water and in the presence of aqueous chlorhexidine and quaternary ammonium compounds and may cause infections in neonates.Shah et al. 1999, Pseudomonas aeruginosa is rarely reported from otogenic abscesses despite its presence in the external ear canal of patients with chronic ear disease, a cause of meningitis and brain abscess in a premature baby, Pseudomonas aeruginosa is rarely reported from otogenic abscesses despite its presence in the external ear canal of patients with chronic ear disease, a cause of meningitis and brain abscess in a premature baby. Gellin et al. 1997,the occurrence of Pseudomonas aeruginosa, is on the increase both in postoperative cases and in injecting drug users; up to 37 percent of isolates . The increase in infections in injecting drug users is represented in most series (Tunkel and Pradhan 2002 ,Nussbaum et al. 1992) and may account for up to 40 percent. The disease is often necrotizing and there is diffuse bilateral consolidation on chest radiograph with occasional multiple abscesses.

Gentamicin and carbenicillin can be effective treatment combination of acute infections. ³ Some B. cepacia genomovars worsen prognosis of CF patients, (Jones et al. 2001). It is intrinsically highly resistant to antibiotics, including those effective against P. aeruginosa but it is usually sensitive to trimethoprim–sulfamethoxazole and chloramphenicol. Wilkinson and Pitt 1995,Stenotrophomonas (Xanthomonas) maltophilia also colonizes with CF patients and can result in disease, it does not adversely affect prognosis. ⁵ These resistance characters may be transferred to other Proteus strains, -to enterobacteria & to Pseudomonas aeruginosa. Unfortunately monotherapy can result in the spread of resistant organisms, e.g. P. aeruginosa, and this has been demonstrated following the use of ceftazidime (Pedersen et al. 1986). Chartrand and Marks 1994,Emergence of P. aeruginosa resistance is also after monotherapy with imipenem and aztreonam. The use of oral quinolones has more flexible management in exacerbations. Ramsey et al. 1999 the use of aerosolized tobramycin given by nebulizer has been shown to eradicate P. aeruginosa and improve pulmonary function tests (Smith et al. 1989), but resistance is a problem.[ Giakkoupi et al. 2003] Carbapenem resistance in Pseudomonas, and some Enterobacteriaceae is becoming an increasing. Two types of class B metallo b-lactamase are prevalent: Verona imipenemase (VIM) and IMP and a VIM-producing P. aeruginosa recently spread among 200 patients in Greece

Resistance pattern observed in Sree Mookambika Institute of Medical Sciences, Kulasekharam which varies from Hospital to Hospital (24-4-2020 to 7-4-2021)

Burkholderia (Pseudomonas) cepacia is a lysine positive, that can be distinguished from Pseudomonas spp opportunistic pathogen in being of cystic fibrosis patients. ³ .Pseudomonads found in bottled waters at densities ranging from 103 to 105 organisms per milliliter reported in some drinking-water supplies include Pseudomonas aeruginosa, Burkholderia cepacia (Pseudomonas cepacia), Pseudomonas fluorescens, Burkholderia mallei (Pseudomonas mallei), Stenotrophomonas maltophilia (Pseudomonas maltophila), Pseudomonas putida, and Comamonas testosterone (Pseudomonas testosteroni),also Pseudomonas stutzeri, Brevundimonas diminuta (Pseudomonas diminuta), and Delftia acidovorans (Pseudomonas acidovorans).Alginate interferes with antibody coating ,inhibits phagocytosis of P. aeruginosa. Pseudomonas elastase increases the permeability of epithelial cells, and induces shedding of epithelial cell-surface heparan sulfate proteoglycans, encouraging colonization.The pigments pyocyanin and 1-hydroxyphenazine , the hemolysin rhamnolipid interferes mucociliary transport .In CF patients produce markedly elevated elastase produced by P. aeruginosa interferes with their activity, so that they are unable to interact with receptor sites on pulmonary macrophages or neutrophils ⁵ encourages persistent colonization.

6] Biofilm:

Biofilms begin with a single bacterium attaching by binary fission and ultimately a glycocalyx for environmental protection. Quorum Sensing is bacteria within a biofilm produce small molecules, such as homoserine lactones, which are taken up by adjacent bacteria and functionally serve as a colony, informing individual bacteria to turn on certain genes at a particular time and these signals are known as quorum sensors. ¹⁷ The genetics of this process are well understood for Pseudomonas aeruginosa, in which the ica gene cluster orchestrates the production of biofilms . Davies et al. 1998 quorum sensing is also likely to be important within biofilms and during biofilm formation . ⁷

Tests: Presumptive tests used for identification of the Pseudomonas spp

Brown's Opacity Tubes: Comparing the turbidity (opacity) of the suspension with the graded turbidities of a series of ten standard tubes (obtainable from, for example, Burroughs Wellcome & Co., London). ¹⁴

Plasma: Oxalate or heparin plasma can be used and not citrated plasma because citrate-utilizing bacteria e.g. , Pseudomonas may cause clotting of the plasma (in tube test). Occasionally, human plasma may contain inhibitory substances which can interfere with coagulase testing can be stored frozen in amounts ready for use. ¹² Pseudomonas organisms may also produce red colonies on XLD . Pseudomonas strains form small green colonies in TCBS agar glucose oxidation positive and glucose fermentation negative, positive cytochrome oxidase test and pigment production : P. aeruginosa produces yellow pyoverdins (fluorescein) and/or pyocyanin (blue aqua pigment). There characteristic grapelike odor of aminoacetophenone as well as growth at 42°C . There are also nonfluorescent Pseudomonas species (P. stutzeri, P. alcaligenes, P. pseudoalcaligenes).⁹ Motility was not studied in any of the species of Pseudomonas, with the exception of P. aeruginosa. Perhaps they are involved in propelling the cell over the surface of solid media (Palleroni et al. 1970; Shinoda and Okamoto 1977 Out of 353 strains, Jessen (1965) found that 19 strains were nonmotile 5[ subsequently acquired] of characteristic of motility. In addition to the polar one P. Stutzeri has lateral flagella of shorter wavelength. ¹⁰ The study by Okhravi et al. 2000 identified , Pseudomonas spp., through detecting bacterial DNA by PCR and by restriction fragment length polymorphism and/or sequencing techniques.

An example of a pigment forming organism is Pseudomonas aeruginosa which gives a yellow-green colour in media such as blood agar and MacConkey agar¹². Pyocyanin, a blue phenazine derivative characteristic of P. aeruginosa, is diffusible and its production can be enhanced by growth in the ‘King A’ medium (King et al. 1954) recognized as the cause of ‘blue pus’ in wounds, named Bacillus pyocyaneus by Gessard the name was changed by S_edillot in 1850 to Pseudomonas pyocyanea, and later called Pseudomonas aeruginosa by Schroeter in 1872 .Chimeric virus-like particles also protected dogs from a lethal challenge with CPV (Langeveld et al. 2001b).

Monoclonal antibodies for outer-membrane proteins have been isolated and characterized by (Hancock et al. 1982). One of the antibodies specific for an antigenic epitope on an outermembrane protein of P. aeruginosa, H2, was recommended as an identification tool for use in clinical laboratories (Mutharia and Hancock 1985) is detected in all 17 serotype strains of the species and, in addition, in other Pseudomonas species that were tested, but not in any species of other genera of aerobic pseudomonads tested.

VACCINE?

A multivalent vaccine based on peptides of CPV, canine distemper virus (CDV), and Pseudomonas aeruginosa could be a commercially viable strategy because they would protect dogs against two important canine diseases and minks against the three major diseases in mink farming.

Pseudomonas answers catalase +oxidase +growth on TSA Pseudomonas on mcConkey agar.

Pseudomonas grown in a cetrimide agar. Pseudomonas in a nutrient agar.

Pseudomonas aeruginosa identified by PNA FISH. PNA FISH is a rapid and highly sensitive and specific fluorescent assay for the detection of Gram‐negative pathogens, including P. aeruginosa. The specimen is placed on a slide and fixed. After fixation, a drop of the probe solution isadded and hybridized. The slide is washed and read using a fluorescent microscope. The red color signifies hybridization with P. aeruginosa.

1. TOPEY WILSON .What’s in a new name? Pg 17

2.ROBERTW. BAUMAN, PH.D MICROBIOLOGY WITH DISEASES BY TAXONOMY ,CHAPTER 20 Pathogenic Gram-Negative Cocci and Bacilli.. Fifth edition global edition,. Pg 630

3. ELSEVIER’S INTEGRATED REVIEW IMMUNOLOGY AND MICROBIOLOGY CHAPTER 12 Clinical Bacteriology, SECOND EDITION ISBN: 978-0-323-07447-6/ 1600 John F. Kennedy Blvd. Ste 1800 Philadelphia, PA 19103-2899 Pg 116

4. ROBERT R. HARR.MEDICAL LABORATORY SCIENCE REVIEW / Chap 7.3— 4th ed. ISBN 978-0-8036-2828-1 — ISBN 0-8036-2828-5 | Nonfermentative Bacilli.Pg 400

5. TOPEY WILSON . Bacterial infections of the lower respiratory tract .Chap 29

6. TOPEY WILSON Clinical features Chap 8

7. TOPEY WILSON Bacterial infections of bones and joints . Chap 5

8. TOPEY WILSON Rapidly growing mycobacteria Chap 17

9. TOPEY WILSON Pseudomonas Chap 2

10.TOPEY WILSON Pseudomonas Chap 4

11.TOPEY WILSON Bacteriology of soils and plants Chap 4

12. MONICA CHEESBROUGH District laboratory practice in tropical countries Cambridge University Press Part 2 Second Edition CHAP 46 The Edinburgh Building, Cambridge CB2 8RU, UK First published in print format ISBN-13 978-0-521-67631-1 ISBN-13 978-0-511-34842-6 © Monica Cheesbrough 2000, 2006 pg 102

13. MONICA CHEESBROUGH MICROBIOLOGICAL TESTS District laboratory practice in tropical countries Cambridge University Press Part 2 Second Edition The Edinburgh Building, Cambridge CB2 8RU, UK First published in print format ISBN-13 978-0-521-67631-1 ISBN-13 978-0-511-34842-6 © Monica Cheesbrough 2000, PG195

14. W. F. HARRIGAN AND MARGARET E. MCCANCE Laboratory Methods in Microbiology Department of BacteriologyDETERMINATION OF THE TOTAL NUMBER OF ORGANISMS IN A SAMPLE, The West of Scotland Agricultural College Auchincruive, Ayr, Scotland Chap 33

15. KEITH STRUTHERS Introduction to Clinical Microbiology Second Edition Chapter 1 CRC Press Taylor & Francis Group 6000 Broken Sound Parkway NW, Suite 300 Boca Raton, FL 33487-2742 © 2017 by Taylor & Francis Group, LLC CRC Press is an imprint of Taylor & Francis Group, an Informa business pg4

16. W. JOHN SPICER . Clinical Microbiology and Infectious Diseases SECOND EDITIONSPECIFIC PATHOGENS BACTERIAElsevier’s Health Sciences Rights Department, 1600 John F. Kennedy Boulevard, Suite 1800, Philadelphia, PA 19103-2899, USA: phone: (+1) 215 239 3804.pg 52

17.JAWETZ, MELNICK, & ADELBERG’S MEDICAL MICROBIOLOGYTWENTY-Eighth Edition . CHAPTER 4 Growth, Survival, and Death of MicroorganismsCopyright © 2019, 2015, 2013, 2010, 2004, 2001, 1995, 1991, 1989 by McGraw-Hill Education ISBN: 978-1-26-001203-3 MHID: 1-26-001203-4.pg58