Pseudomonas Genus of Pathogens - Literature review Example

Pseudomonas Isolation: Literature Review Student Name Tutor Course Date Pseudomonas Isolation LITERATURE REVIEW Introduction Medical attention has long been drawn to Pseudomonas genus of pathogens which take advantage of situations and cause infections in case an individual immune system is not strong enough. Some clinical infections arise as a result of pseudomonas contaminating equipment, fluids or items used during treatments. According to Madigan and Martinko (2005), bacteria under this category display a high degree of metabolic diversity capable of inhabit a wide range of areas. Members of the Pseudomonas group are known to resist antibiotics over time. This is majorly as a result of diversification in their metabolism. The bacteria’ gram negative aerobic bacillus gives them a more advantage for adaption since they are also able to survive in almost all environments. Vegetation, water and soil are the main environments that providing an efficient breeding place for pseudomonas bacteria. Bacterial isolation involves sequestration or separation of bacterium either by their characteristics species or their breeding sites conducted in the labs or environments where the bacteria breed. Traditional microbiological procedures have widely been used to conduct isolation of the microorganisms. The traditional techniques include metabolic reactions procedures, gram stain reaction and morphology. Physical isolation of bacterium is easily conducted by the medics in laboratories to identify what bacterium is responsible for the given disease in a given patient. Isolation can be important to separate these bacteria since they do not occur in lone pairs but within communities of other micro-organisms like them within animal or human body and in the environments where they breed. Roles of bacteria in disease transmission also occur in communities of the same within the body. Identification of bacterial characteristics of a given bacteria, suitable habitants for their survival and development mechanisms including survival adaptations facilitate the isolation process and makes it successful. Once a patient is already ill and their immune system is weak, these bacteria act as opportunistic pathogens. That is, they take advantage of a weak immune system and not an immune system which is strong enough to fight them. According to Hertveldt (2005), this group of bacteria easily causes outbreaks if they are not well controlled and end up causing infections in a wide range of niches. He states that, biofilm slime matrix growth is the bacteria adaptation mechanism which enables them to survive in the many environments they inhabit. Phagocytosis and other activities of that kind are prevented from affecting the bacteria by the exopolysaccharide which surrounds the micro colonies. Although pseudomonas bacteria are pathogens, they possess both positive and negative qualities making them both helpful and also harmful. This paper reviews literatures from scholars on pseudomonas and the isolation of bacteria which fall in that group discussing positive and negative effects in relation to isolations and the environment. Methods of isolation, characteristics of the bacteria and medium of breading are also explored in the pseudomonas literature. Pseudomonas and their isolation Pseudomonas is likely to survive in water for a long time because it does not need too much nutrients to survive in water, but even under minimal levels of nutrients they still live. They are mostly found in animals, plants, faeces and human oropharynx since they provide a better breeding environment for survival and reproduction. Clinical infections that result from pseudomonas infections arise from suitable breeding environment for the bacteria such as aqueous solutions which are contaminated. Dialysis fluids, eye drops, equipment, antiseptics, soaps and tap water are the solutions that facilitate pseudomonas breeding and bring about infection in a clinical environment (Krueger and Sheikh, 1987). Invasive devices that are used on patients are also a medium that providing breeding sites for pseudomonas thus causing clinical infections, such as catheters, endotracheal tubes and ventilators. Isolation of bacterium physically in laboratories also involves knowledge of the microorganisms’ survival mechanism. These include the maximum oxygen requirement for their survival, optimum temperature that facilitate their growth and ideal nutritional requirements. However, some bacteria are not easily cultured in the lab. Laboratory isolation procedures include using an agar plate to streak isolation and employing the pour plate method. Use of the agar plate in laboratories for streaking isolation of the bacteria implicates continuous attenuation of organisms. This is done up until the cells have attained a lower density to the extent that individual cells became isolated physically and spatially. At this point, individual colonies can be identified. Pour plate method includes sufficient dilution of the microorganisms’ sample which is then poured into molten agar that has been cooled before the mixture is transferred into a petri dish. However, there can be challenges involved for example, the melted agar can be at very high temperatures and in turn kill all sample bacteria. Very cool agar can also result to inefficiency since it results to a bigger lump of sample in the petri dish. According to Northeast laboratory services, (2009), the agar technique for streaking is easy to comprehend and fast enough compared to the pour plate method since it produces single colonies on the agar surface. Isolation of the bacteria can be conducted using asparagine broth where soil is placed in sufficient amount of the chemical like in the experiment conducted by Al-Hinai et al (2010). Asparagine broth is used in the procedure to improve growth of the pseudomonas bacteria through the asparagine L-monohydrate compound contained in it. This step is followed by a 48 hours incubating the sample at a temperature of 37 degree Celsius. This should be shaken at 200 rpm so as to provide airing for bacteria. The process results to a ring of bacteria suspended which is then lined on asparagine plates with agar, also known as Oxoid, UK and maintained at 37 degree Celsius to the point where colonies are formed. After the colonies form, they are transferred to another new asparagine plates under the different morphological features of colony such as their size, their color or shapes. Identified qualities from isolation Isolation of pseudomonas bacteria has long revealed their importance in reducing damping-off disease caused by the pythium pathogen in cucumber. Studies have indicated that fungal bio control agents can be used to inhibit this damping off on Oman’s most productive crop, the cucumber and also sugar beet. Al-Hinai et al (2010) in their article on isolating and characterizing pseudomonas aeruginosa employ aggressive activities against pythium aphanidermatum. The authors conduct a greenhouse bacterial isolation and characterization exercise on the soils. From the study, pseudomonas aeruginosa are identified to be the bacterial microorganisms which suppress damping off of pythium by cucumber. The study was aimed at identifying the bacteria responsible for suppressing damping-off of pythium by cucumber. Identification of the isolates in terms of their species was conducted using API20NE and a number of 16S ribosomal RNA. Pseudomonas bacteria have their optimum growth rated at a temperature of 30 degree Celsius while they display active contentious behavior at the temperature of 37 degree Celsius. This category of pseudomonas bacterial display unresponsiveness to salinity environments which are increased to high levels of 10 dS m-1. Their growth and antagonistic activity does not change as a result of change in salinity to the highest level. This therefore displays high levels of tolerance of pseudomonas bacteria. On the other side, suppression of pythium damping off was evident at a temperature of 28 degree Celsius according to the authors’ experiments but did not have an effect on the growth of greenhouse seedlings of cucumber. The authors state that, the pseudomonas species are used in controlling many other diseases in crops like sugar beet other than cucumber. It is therefore possible that pseudomonas species aeruginosa exists in the soil of a greenhouse and can survive in such environments. Pseudomonas aeruginosa and Aeromonas can also be isolated using membrane filtration from drinking water. The membrane filters used to isolate bacteria are positioned on a solid surface with potassium sulphate and magnesium chloride which facilitates formation of pigment. Nalidixic acid and cetyl trimethylammonium bromide are added to the solid surface where membrane filters have been placed so as to provide selectivity ability to the medium. Isolates are subjected to milk agar compound which is complemented with a compound cetyl trimethylammonium bromide (CMA). Alternatively, Mueller Hinton agar can also be sub-cultured and phenanthroline impregnated discs added to achieve the same purpose of casein hydrolysis. Since drinking water is considered as having no nutrients, it is expected that pseudomonas pathogens do not survive in such waters. However, normal intestinal systems of animals and humans are found to have traces of such pathogens. According to Environmental agency (2010), patients suffering from other infections like ear infections, septicaemia, urinary tract, skin or respiratory infections after conducting other diseases are likely to have been infected by the pseudomonas pathogens. Testing such infections and isolating the pathogens is important. The author states that, swimming pools, and waters at spas are likely to have most numbers of the microorganisms. This provides a medium for follicular dermatitis or ear infections when people immerse in the waters to swim. However, the micro-organisms are significant in treating water and water supply and are important in serving their purpose due to their resistance feature to antibiotics. In isolating these bacteria, control measures can be taken to confirm bacteria colonies as indicated in the diagram below: Role of bacteria and bacteria isolation in disease Bacteria play a major role in causing diseases and causing infections to patients. Since the existence of the bacteria in human body and that of animals occur in communities, treatment of bacterial diseases requires isolation of the single bacterium species. Medics identify the responsible bacteria for a given disease in patients through isolation processes of bacterium into their individual species. In this case, bacteria can be cultured in the hospital labs or clinic for physical isolation. Incidences of bacteria in clinical environments occur on surfaces, inserted devices and water, thus causing infections to patients who are weak. An example is the use of catheters in hospitals which are inserted in a patient’s body. Infections related to catheters are known to contribute about 31% to hospital related infections. These infections can be prevented and controlled by standardizing the insertion kits and coating the catheters with linings that will discourage accumulation of bacterial elements. Johnson et al. (2012) state that urinary tract infections is one of the infections that occur as a result of exposure to the hospital environment. This is associated with the accumulation of microorganisms in the area which such devices are inserted within the body. Diseases contracted by patients in this case are referred to acquire diseases such as growth of a biofilm. Various species of bacteria can develop around the catheter after it is inserted in the human body. The bacteria are likely to discharge polysaccharide components. The biofilm created afterwards causes infection to the body (Du Plessis, 2011). The article written by Curtis (2008) involves prevention measures that can be undertaken to prevent nosocomial infections in hospitals which are majorly caused by bacteria using non pharmacological intermediations. The study involves a thirteen years research using bibliographic data of research journals, medical websites and hospitals. The control interventions as stated by the article include better nutrition to patients, ensuring that ventilations are well managed and ensuring cleanliness on surfaces and hands. The author also states that having appropriate number of nurses to attend to patients is also another control measure. Another measure mentioned is ensuring that air filters used are highly efficient and particulate to reduce the rates of nosocomial infections and using catheters which are coated for central venous and urinary operations. The article illustrates that employing several procedures and strategies in controlling infection at hospitals has a greater impact in controlling the hospital based infections on patients thus reducing mortality rate and cut on the increasing rate of morbidity (Dancer, 2009). Pseudomonas aeruginosa infects eyes, wounds, ears or burns and are well known as opportunistic pathogen. For this reason, it is widely known to cause most nosocomial infections acquired by patients. According to Aumedia (2011), pseudomonas aeruginosa are known to cause infection mostly to patients who are undergoing therapy of the antibiotics. This group of pseudomonas bacteria can be isolated from other groups of pseudomonas using agar by adding glycerol and is accomplished through formation of pigment. After incubation, the bacteria is expected that it turns green if it is incubated for about eighteen hours and turn blue-green if incubated to about 24 to 28 hours. Nitrofurazone is one of the components used in coating catheters as a measure of infection control. Nitrofurazone is an agent that kills bacteria (bactericidal) used as an antibiotic or disinfectant. It does not kill any other components other than bacteria and appears as a pale yellow in color when in liquid form which is also crystalline. Its chemical structure is made up of carbon, hydrogen, nitrogen and oxygen components. Coating catheters with the compound kills bacteria and prevents their accumulation on the surfaces. This as a result works as an infection control measure. Another coating component used to control infections in hospital environments is silver. Silver alloy contains its ion (Ag+) which sufficiently kills bacteria and other microorganisms. Other than the ion, silver and silver Nano components also kill bacteria and prevent accumulation on surfaces thus controlling any infections likely to be caused. Siegel et al (2007) provides a precaution guideline that can be used to prevent infectious agents from being transmitted in the healthcare environments. The authors state that healthcare settings might move from the primary acute kind of care and also embrace other measures like long-term cares, ambulatory care or home care. This is in order to suit specific needs settings while applying standards recommended by the infection control practice principles. Healthcare associated infection is a term that explains evolution of the term nosocomial infection due to changing practices in the healthcare infection environment. The authors state that, modification of the previous guidelines and adjustment of the same were as a result of emerging species of pathogens like in pseudomonas group, SAR-COV which causes severe respiratory syndrome and evolving ones like difficile, noroviruses. Antibiotics are criticized by researchers stating that if non pharmacological techniques are well used, patients will not have to be supplied with a couple of antibiotics for treatment of clinical bacterial infections especially those resulting from opportunistic pathogens. Patients have to leave on antibiotics to prevent infections and treat them. Curtis (2008) suggests the non-pharmacological techniques since excessive use of antibiotics control develops a resistance to the same in patient’s bodies. In this case, further use of the drugs will not help such a patient. Low antibiotic usage increases the effectiveness of the drug in the body (Helder, 2012). Isolation of bacteria through classification and treatment of the specific species is the best way of dealing with such infections. According to Siegel et al (2007), other conditions like respiratory hygiene etiquette in coughing and employing safe practices during injection such as use of masks during highly risky procedures concerning punctures of the spinal cord have been affirmed to reduce such infections to a great deal. Spreading of the SARS coronavirus and other pseudomonas infections have been speeded up by failure in control measures during break-ups among personnel in the healthcare, visitors and to patients. Pseudomonas is identified by the authors to have resistance to environmental control measures and drugs administered to kill them and prevent further infections caused by the pathogens. This is due to mutation and changing metabolism of the micro-organisms. They are classified under category of multidrug resistant organisms, also known as MDROs. MDROs are resistant not only to a given class but a number of antimicrobial agents, especially chemical agents. Isolation and understanding the behavior of such pathogens is of rising importance in the clinical field. There is a high possibility of emergence and transmission of these pathogens among patients even staff. However, these can also be controlled by application of committed measures by the administration through full involvement. The administration is likely to achieve controls through activities like sufficient communication systems, improving the processes of performance, training and education personnel responsible for the healthcare. Siegel et al (2007) suggest that, medics should also try to survey individually targeted MDROs and employ environmental precautious care and also employ decolonization therapy where possible. Pathogens classified as MDROs require that control measures are changed against their metabolism and mutation systems for effective control of infections. Disinfection of clippers during surgery is an example of control measure used to control cross infection between patients. Clipping involves using clippers which fine teeth to remove hair around surgical areas. The hair is cut leaving it at a length of only 1mm from the skin surface. The procedure also involves disinfection of clipper heads or disposal of the tools after using them on one patient before attending to another. It reduce chances of cross infection between patients. Shaving is the most common technique used in preoperative hair removal procedures. It is cheaper compared to other hair removal methods involving razors which are fixed with sharp blades at the heads. It is then drawn over the skin of a patient cutting hair and leaving it short and close to the skin surface (Hemingway, 2007). There adverse exposure to microorganisms in the health care environment. Moist organic environments are likely to be more exposed than dry environments. According to WHO (2002), it is the duty of workers in the social and healthcare facilities to practice infection control management to ensure the safety of patients. Antibiotics and diagnostic approaches can be used to detect and manage infections before they occur. Issues like preoperative hair removal to reduce surgical infection as a result of surgical site infections and use of coated catheters to impend micro-organic infection in urinary tract prevent micro-organic infections in clinical environments. Conclusion Pseudomonas bacteria are an opportunistic pathogen that takes advantage of a weak immune system to cause infection to patients. However, the bacteria also have an advantage other than infection both in human or animal body and plants such as cucumber and sugar beet which experience damping-off, and can only be solved using deposits of the bacteria colonies. Isolation of the bacteria involves use of different methods like use of agar plate, pour method and others. Agar method is considered simpler and provides quick comprehension. Matrialscan also be suplimented to achieve the same results. Pseudomonas is also known to contribute a big deal to nosocomial infections on patients which are easily controlled using pharmacological and non-pharmacological techniques. Pharmacological techniques will include administration of antibiotic drugs. Although some researchers considers both methods as being efficient and appropriate, some criticize use of antibiotics since they are known into cause resistance in the patient’s bodies if used frequently and also resistance by the pathogens. Bibliography Madigan, M and Martinko, 2005, Brock bology of micro-organisms. Prentice Hall. Johnson, J.R., Johnston, B. & Kuskowsk, M.A. (2012). In Vitro Comparison of Nitrofurazone- and Silver Alloy-Coated Foley Catheters for Contact-Dependent and Diffusible Inhibition of Urinary Tract Infection-Associated Microorganisms. Antimicrob Agents Chemother. 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