Treating the Bacterial Disease - Essay Example

LABORATORY REPORT Treatment of bacterial diseases requires specific medication that targets thepathogen to avoid development of resistance. Therefore, diagnosis of enteric and other pathogens require precise identification process that narrows down to the characteristics distinguishing one pathogen from the rest. The study aimed to identify pathogen using a series of simple biochemical tests like catalyse, oxidases and the grams staining technique as well as the morphological characteristics of the colony to identify the mysterious bacterial pathogens. These strategies were used alongside the cellular arrangement. Besides, the experiment studied resistance patterns of the pathogens against common antibiotics. The study found that all the bacterial pathogens were found to be susceptible to chloramphenicol. However, only staphylococcus aureus was found resistant to streptomycin, the rest were susceptible. For the ampicilin resistance, only, Pseudomonas fluorescens was resistance. Testing the resistance for penicillin depicted that booth Escherichia coli and Pseudomonas fluorescens were resistance to penicillin. Pseudomonas fluorescens was distinguished from other rod shaped single cells arrangement by the positive oxidase test. The rod shaped single cell arrangement that produced gas for glucose, fructose, and lactose fermentation was identified as that Escherichia coli. These findings shows that pathogens exhibits different morphological a biochemical tests. Combination of these tests helps in the specific identification of the pathogens. 1.0 Introduction Recent development of resistance by the enteric and environmental pathogens is a dangerous phenomenon because scientist would not be in a position of using most of the current broad spectrum in the treatment of pathogens (Hariharan et al., 2015). The main problem occurs when there are several bacterial pathogens in a particular infection. Such scenarios makes it difficult to target a specific pathogen because one remedy may not be effective for all the bacterial pathogens since some of the bacterial have developed resistance to the antibiotics (Fischbach and Walsh, 2009). However, the most important strategy for combating the increasing cases of bacterial infection is the appropriate identification regimes. Effective identification requires the use of specific strategies like the use of a polymerase chain reaction. Using the PCR helps in the identification of the genome of the pathogen hence increasing the likelihood of specific identification (Piddock, 2014; Silver, 2011). However, not all laboratory setting and health facilities can afford the procedure. Therefore, the best alternative is the combination of simple yet effective strategies that identifies pathogens based on the specific characteristics. The identification of pathogens is important before doing the antibiotic resistance tests that would help identify the most appropriate treatment regime for use in case of coming up with appropriate treatment method to avoid development of resistance to the antibiotic (Piddock, 2014; Silver, 2011). The experiment aimed to identify pathogens based on their morphology, biochemical characteristics, and cellular arrangement before studying their resistance to specific antibodies. 2. Materials and methods 2.1 Chemicals and reagents Four antibiotics namely penicillin, chloramphenicol, streptomycin, and ampicilin were selected for testing antibiotic resistance. The nutrient agar and agar disk were used for antibiotic resistance study. Gram stain reagent and staining rack were used for the preparation of the studies of cell morphology. The microscope was used for observing the stained slides that were covered with cover slips 2.1 Disk diffusion assay The disk diffusion was used for the antibiotic resistance studies. The agar was prepared and plated with specific bacteria. A disk was made in the agar and antibiotic poured. The experiment was incubated and checked after 18hrs. The diameter with no growth referred to as the inhibition zone was determined using the diameter measurement. Measurement of the diameter less than 10mm was reported as resistance development while diameters greater than or equals to 10mm signified susceptibility 2.2 Biochemical and morphological tests Tests like oxidase, catalase test, fermentation of sugar, lactose, and fructose were performed to assess the biochemical characteristics of the pathogens. A colony of specific bacteria colony was used to test the specific test as previously described (Silver, 2011). 3. Results 3.1 Antibiotic resistance All the bacteria pathogens were found to be susceptible to chloramphenicol. However, only staphylococcus aureus was found resistant to streptomycin, the rest were susceptible. For the ampicilin resistance, only, Pseudomonas fluorescens was resistance. Testing the resistance for penicillin depicted that booth Escherichia coli and Pseudomonas fluorescens were resistance to penicillin. 3.2 Morphology and biochemical Pseudomonas fluorescens was distinguished from other rod shaped single cells arrangement by the positive oxidase test. The rod shaped single cell arrangement that produced gas for glucose, fructose, and lactose fermentation was identified as that Escherichia coli. The other rod shaped pairs or short chains of cell arrangement that was positive for grams stain was identified as Bacillus megaterium. The other positive grams reaction for the cocci irregular clusters of cell arrangement was identified as staphylococcus aureus because it fermented glucose, fructose, and lactose. The unknown culture was rod shaped single cell arranges grams negative, with positive catalyse, glucose, fructose, and lactose fermentation. The other rod shaped pairs or short chains of cell arrangement that was positive for grams stain was identified as Bacillus megaterium. The other positive grams reaction for the cocci irregular clusters of cell arrangement was identified as staphylococcus aureus because it fermented glucose, fructose, and lactose. The unknown culture was rod shaped single cell arranges grams negative, with positive catalyse, glucose, fructose, and lactose fermentation. To identify the pathogen simply complete the blanks in the table below and compare your results to the diagnostic characteristics provided. Table 3.1:Table of characteristics for interpretation of biochemical bacterial identification tests. Simply match the characteristics. Name of bacterial species Penicillin (P) Ampicillin (AMP) Streptomycin (S) Chloramphenicol (C) Pseudomonas fluorescens R R S S Escherichia coli R S S S Staphylococcus aureus S S R S Bacillus megaterium S S S S Number or letter of unknown culture is: All the organisms reported positive to Catalase test, oxidase test was negative in all except the pseudomonas Discussions All the bacterial pathogens were found to be susceptible to chloramphenicol, which suggested the need to use this antibiotic as the broad-spectrum choice for the treatment of all the pathogens. However, only staphylococcus aureus was found resistant to streptomycin, which suggested the need to avoid using this antibiotic against the bacteria owing to the potential of resistance development. For the ampicilin resistance, only, Pseudomonas fluorescens was resistance suggesting that ampicillin is better alternative for all the pathogens with the exception of this pathogen. Testing the resistance for penicillin depicted that both Escherichia coli and Pseudomonas fluorescens were resistance to penicillin. One strategy is the use of grams staining to group the pathogens in two broad categories of positive or negative (Fischbach and Walsh, 2009). The staining also helps in the identification of their cellular characteristics like shape and cell arrangement. Other strategies involve the use of biochemical tests that studies characteristic of the pathogens to procedures like oxidase, catalase, and fermentation of carbon molecules. Sugar fermentation is different between the organisms because these pathogens have different capacities of using glucose as source of energy. E. Coli ferment glucose to produce gas as opposed to other species REFERENCES Fischbach, M.A & Walsh, C.T. (2009). Antibiotics for Emerging Pathogens, Science, 325(5944): 1089-1093 Hariharan P, Bharani T, Franklyne JS, Biswas P, Solanki SS, Paul-Satyaseela M (2015). Antibiotic susceptibility pattern of Enterobacteriaceae and non-fermenter Gram-negative clinical isolates of microbial resource orchid. J Nat Sci Biol Med. 6(1):198- 201. Lewis, K. (2013). Platforms for antibiotic discovery, Nature Reviews Drug Discovery, 12(May): 371-387 Piddock, L.J.V. (2014). Understanding the basis of antibiotic resistance: a platform for drug discovery, Microbiology. 160(Pt11): 2366-2373 Silver, L.L (2011). Challenges of Antibacterial Discovery, Clinical Microbiology Review, 24(1): 71-109 Read More