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Posted: April 28th, 2022
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INTRODUCTION.
The aims of the investigation are to identify the effect of bacteria, for example, what effect can Serratia marcescens pilli on red blood cells gives when mixed with molecular inhibitor. Also, to examine how pathogenic bacteria for example Clostridium perfringes produce toxin and what effect does toxin have on cells. In addition to the above, to examine the microbial resistance by using four different antibiotics by BASC method this will guide us to know the sensitivity and resistance to particular type of antibiotics.
Pathogenicity is the capability of microorganism to cause disease. According to Madigan et al (2009) the favarouble environment for growth of microorganism is live in or on human body and cause disease. Bacteria have protein filaments called pili which bind to the epithelial cells and cause infection.Pili are much more common in Gram negative bacteria. In our study we used Gram negative pathogenic organism Serratia marcescens which can cause disease to human also Serratia marcescens genome has pili on cell surface for adherence to the epithelial cells and cause infection. In our investigation we use this bacteria suspension to agglutinate with erythrocytes by examined the activity of pili in the presence of molecular inhibitor. Also we investigate the pathogenicity of bacteria by analysing the toxin they produce. In addition we study on microbial resistance to different antibiotics to know the sensitivity and resistance to the drugs. Since 1950s, pathogenic microorganisms have developed the ability to resist some antimicrobial chemotherapy agent. The earliest widely used of chemotherapy agents were amongst penicillin and sulphur drugs. However, these drugs are no longer commonly used as they were back in 1950s, this due to majority of pathogens have obtained some resistance.( Madigan et al 2009).Furthermore, at present some strains of pathogens become resistance to almost all antimicrobial agent, such as Staphylococcus aureus, Mycobacteriun tuberculosis and Candida albicans which are not treatable. An experiment of antimicrobial resistance can be carry out to test some antibiotics which are Cephalosporin’s group, against five strains
RESULTS : DILUTION Table 1
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DILUTION |
TEST |
CONTROL |
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Positive within 20sec |
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10-2 |
Positive within 30sec |
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10-3 |
Positive within 30 sec |
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10-4 |
Weak + within 30 sec Can You Follow My Professors Unique Guidelines?Absolutely! Share your professors rubric or specific instructions, and we will tailor the paper to meet every detail, no matter how unique. Our writers thrive on precision and customization. We have experience with unconventional requirements and specialized formatting across all disciplines. Upload your guidelines in the order form for best results. Research study bay professionals excel at interpreting and executing complex instructor specifications. |
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10-5 |
Weak within 30sec |
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The table above shows the results which obtain after bacteria dilution were tested against erythrocytes.
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PILI RESULTS: Table 2
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SUGARS |
TEST |
CONTROL How Do I Specify My Requirements?Use our order form to detail your topic, style, and any specific instructions, or upload files for your writer to follow precisely. Clear instructions ensure a perfect paper every time. The more details you provide, the better we can match your expectations and academic goals. Our support team can guide you if needed. Essay writer assignments benefit from comprehensive requirement descriptions in the initial order. |
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Glucose |
Positive after 15 seconds |
Positive after 5sec |
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Fructose Are Your Writers Native English Speakers?Many are native speakers, and all are fluent, passing strict language tests to ensure clear, professional writing for your papers. This guarantees polished, high-quality work. Our language proficiency standards ensure flawless grammar and natural academic expression. You can request a native speaker if preferred. My assignment help comes from linguistically skilled professionals who master academic English. |
Positive 35 seconds |
– |
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Mannose |
Negative Can You Help with Literature Reviews?Definitely! Our writers craft comprehensive literature reviews, synthesizing sources to support your research with clarity and depth. We ensure every source is relevant and properly cited. Literature reviews require critical analysis and thematic organization that our experts deliver consistently. Just provide your topic or sources to get started. Research essay service includes meticulous literature synthesis for scholarly credibility. |
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Table 2 shown the results when Serratia marcescens tested with erythrocytes to check for their ability to agglutinate in the presence of glucose, fructose, and mannose.
FOUR BACTERIA SUSPENSION Table 3
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MANNOSE |
WITHOUT MANNOSE |
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Bacteria |
Agglutination How Do You Handle Group Projects?We can write sections for group projects, ensuring consistency with your teams style and integrating seamlessly with other parts. Our writers collaborate to match your groups vision. Coordinated group work requires attention to unified tone and formatting standards. Upload any shared guidelines to keep everything aligned. Paper writing for collaborative assignments maintains cohesion across all contributed sections. |
Time |
Agglutination |
Time |
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310258 |
Positive Can You Assist with Grant Proposals?Yes, our writers have experience crafting compelling grant proposals, tailored to meet funding requirements with persuasive arguments. We help you make a strong case for funding. Successful proposals require strategic narrative and evidence-based justification that we specialize in. Provide your guidelines, and we will deliver a winning proposal. Assignment writer expertise extends to professional grant writing with proven success rates. |
15 sec |
Strong Positive |
10 sec |
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310217 |
Negative What Is Your Turnaround for Editing?Editing typically takes 24-48 hours, depending on paper length, with rush options available for faster delivery. We polish your work to perfection quickly. Our editors focus on enhancing academic tone while maintaining your original ideas and arguments. Upload your draft to start the process today. Assessment help through editing services delivers professional refinement on tight schedules. |
30 sec |
Weak Positive |
25 sec |
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Negative |
30 sec |
Weak Positive |
20 sec |
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706498 How Do You Ensure Writer Expertise?Our writers are vetted for degrees and expertise, with ongoing evaluations to maintain high standards across all subjects. This ensures your paper is handled by a true professional. Regular quality audits and performance reviews keep our team at peak excellence. Trust us to match you with the right expert. Research study bay maintains rigorous quality control through continuous writer assessment. |
Negative |
30 sec |
Negative |
30 sec |
Table 3. Among these four bacteria only one resulted in marked agglutination after 15 seconds in the presence of mannose and strong agglutination observed within 10 seconds without mannose as shown on the table
Toxin test results Table 4
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Lecithinase Can I Request Unlimited Revisions?We offer free revisions within 7 days; additional revisions are available at a small cost if needed beyond that period. Your satisfaction is our goal. Most students find our first delivery meets expectations, but we are flexible for perfectionism. Contact support to discuss any extra changes you need. Ace tutors remain committed to perfecting your work through reasonable revision requests. |
Result |
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Clostridium perfringens |
Strong positive(+++) |
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Staphylococcus epidermidis |
Negative |
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DNAase What Is Included in Your Outlines?Our outlines detail structure, key points, and sources, giving you a clear roadmap for your paper before writing begins. They are designed to keep your project focused and on track. A well-crafted outline serves as both planning tool and progress checkpoint. Request an outline with your order for added clarity. Essay writer planning includes comprehensive outlines that guide the entire writing process. |
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Staphylococcus aureus |
Positive |
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Staphylococcus epidermidis |
– |
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Haemolysis |
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Streptococcus pyogenes |
Positive |
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Streptococcus sanguis Can You Handle Technical Writing?Yes, our writers excel in technical fields like engineering or IT, producing precise, well-documented papers for specialized topics. We ensure accuracy and clarity in every technical detail. Complex technical concepts are explained clearly without sacrificing academic depth. Share your requirements for a tailored result. My assignment help covers specialized technical subjects with field-specific expertise. |
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The table above showed the production of toxin during bacteria growth.
ANTIMICROBIAL SUSCEPTIBILITY RESULTS Table 5
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SAMPLE |
CEFALAXIN |
CEFUROXIMINE |
CEFOTAXAMINE |
CEFEPIME |
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Control |
20/22 How Do You Support ESL Students?We tailor papers to match your language proficiency, ensuring clarity and academic quality while supporting non-native English speakers. Our writers make complex ideas accessible. ESL students benefit from our culturally aware approach and clear academic language. Let us know your needs for a perfect fit. Research essay service accommodates international students with language-sensitive academic support. |
32/30 |
45/40 |
45/44 |
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310258mm |
0 Can You Meet Tight Formatting Rules?Absolutely! We adhere to any formatting guidelines, from margin sizes to heading styles, ensuring your paper meets all requirements. Precision is our promise. Even the most detailed formatting specifications are followed exactly as instructed. Upload your formatting guide to get it right. Essay helper professionals master intricate formatting requirements across all citation styles. |
17/18 |
30/32 |
35/44 |
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310258 S or R |
R What Happens If I Am Not Satisfied?If you are not happy, we offer free revisions within 7 days or a refund review to ensure you are fully satisfied with the outcome. Your peace of mind matters to us. We work collaboratively to address concerns and find solutions that meet your academic needs. Reach out to our support team to resolve any issues. Paper writing satisfaction remains guaranteed through our responsive resolution process. |
R |
R |
S |
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310217mm Can You Help with Annotated Bibliographies?Yes, we create detailed annotated bibliographies, summarizing sources and aligning with your citation style for academic precision. Each entry is concise and relevant. Annotated bibliographies require critical evaluation of sources alongside proper documentation. Provide your sources or let us find them for you. Assignment writer services include expertly crafted annotated bibliographies for research projects. |
0 |
0 |
0 |
0 |
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310217 S or R |
R What Is the Shortest Deadline You Offer?We can deliver quality papers in as little as 3 hours for urgent needs, with dedicated writers ensuring no drop in quality. Perfect for last-minute assignments. Our express service combines speed with excellence for emergency academic situations. Chat with support to confirm timing for your task. Assessment help remains available even for the most urgent deadlines without quality compromise. |
R |
R |
R |
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704476/2mm |
20/21 |
26/20 |
34/36 |
30/35 |
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704476/2 |
S |
S |
S |
S |
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706498mm |
18/22 |
19/17 |
34/34 |
34/36 |
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706498 S or R |
S |
R |
S |
S |
The disc of antibiotics show susceptibility is determined by measuring the diameter of the zone growth of bacteria inhibition. In the table shown above, the bacterium is susceptible to most antibiotics.
DISCUSSION
Five organisms including Serrecia marscence and control were tested for the presence and absence of pili. These organisms were tested with erythrocyte to assess their ability to agglutinate in the presence and absence of different types of sugars. Serretia marscence is Gram negative organism, its genome code for pili on their cell surface because pilli are protein found on the cell surface of bacteria cell. Pilus is a contributing factor of microbial adherence to host epithelial cells. ( I. E. Salit,$ and E. C. Gotschlich) There are there are 2 classes of pili, these are mannose sensitive (MS) and mannose resistant (MR) of adhension One class is mannose resistant pili is capable to agglutinate erythrocyte in the presence of D-mannose known as type non-type 1 pili and second class mannose sensitivite pili adherence linked with mannose sensitive haemagglutination of chicken and guinea-pig erythrocytes and also known as type 1 pili (A. Hejaz and F. R. Falkiner 1997) .In addition Dhakal 2009 said Gram negative organism encoded pili and show significant role in cause infection of some of those of Gram negative bacteria. .
Agglutination test of bacteria were carried out on together with the control as shown on table 1, almost each bacteria dilution after gentle mixing with erythrocytes together with PBS on a slide, result in clear agglutination within 30 seconds on101 to 103 but weak positive observed on104 to 105 within 30 seconds.
On table 2 show the result from the activity of Serratia marcescens pili by means of molecular inhibitor for example mannose, glucose and fructose. Mannose inhibited the agglutination since have the ability of preventing the adhesion of bacteria pili on to the cells. Therefore Serratia marcescens has type 1 pili since it is mannose sensitive, mannose prevents the adherence by protecting cell surface receptors, therefore inactivates the bacteria pili to adhere to the red cells. In place of glucose agglutination occurs in 15 seconds and fructose was showing agglutination within 35 seconds therefore glucose and fructose has no influence to act as inhibitor. Both glucose and fructose are able to absorbed with erythrocytes but unable to stop the adherence. Additionally, agglutination was much slower in control in the presence of glucose and fructose.
Clostridium perfiringes break down the lecithin caused halo around the streak on the egg york agar because egg york contain a lot of lecithin which make easy for detecting lesithinase activity. This lesithinase activity is produce by Clostridium perfringes which cause the breakdown of the lecithin in the cell membrane. Clostridium perfringens contains two main toxins which are alpha-toxins phospholipase and sphingomyelinase but the key toxin is alpha toxins which play main role in the pathogenicity of Clostridium perfringes. As it also contain lecithanase activity as target cell membrane. (O’Brien, 2004) Staphylococcus epidermis does not breakdown the lecithin since does not have toxin contain enzymes to breakdown the lecithin (www1)
In DNase plate two bacteria were tested for DNase reaction, these are Staphylococcus aureus and Staphylococcus epidermins where Staphylococcus aureus produce hallow or clear zone around DNase after covered and acidified with 1NHCL that indicate positive but no precipitates are produced by Staphylococcus epidermis means DNase negative. DNAse is suitable to differentiate Staphylococcus aureus which are coagulase positive from other Staphylococcus which are coagulase negative.(www3)
A simple analysis to investigate the haemolysis of red blood cells used in identification of Streptococcus pyogens and Streptococcus sanguis reaction on blood agar. Hemolytic type reactions used to categorize strains of Streptococci, the hemolysis that is related with extensive lysis of erythrocytes surrounding the colony is known as β-Hemolysis while hemolysis associated with lessening of erythrocytes and produce greening hemolysis is known as α-hemolysis (www2). Result on table 4 show there was a wide haemolysis around the Streptococcus pyogens colony known as beta haemolysis. Streptococcus .pyogens belongs to beta haemolytic group and produce virulent factors such as pyrogenic toxin which play role in haemolysis and produce a clear zone (www 2). There was no haemolysis around the Streptococcus.sanguinis which means it would be simple to investigate these bacteria through their toxins. Especially that S.sanguinis is partially destructing the erythrocytes and belongs to alpha haemolytic group.
Four bacteria were tested according to BSAC method to investigate antimicrobial susceptibility. Cephalosporin’s group was used and cephalosporin is a group of Beta-lactamases antibiotics. The 1stgeneration cephalosporin antibiotics have much effect with gram positive bacteria and less effect against gram negative bacteria. However gram negative bacteria are more effective to 3rd generation and 2nd generation are more active than1st generation.
The table 5 show the bacteria strain 704476/2 was susceptible to all four antibiotics. This suggest that this bacteria indicate have not got any resistance genes in its plasmid and furthermore all four antibiotics can treat the patient infected with this strain as showing on above table. Strain 310217 was resistant to all antibiotics including all three Cephalosporin group, this indicate that this bacteria has resistance gene, so this strain has to be treated with other antibiotics. In addition bacteria strain 706498 only resistant to one antibiotic and susceptible to the remaining three antibiotics while bacteria strain 310258 was resistant to three antibiotics but susceptible to one antibiotic. There is slightly difference that affected my result if I compare my control test with control chart this may be due to human error which can be caused by inoculation, because inoculum size is the most important factor affecting result, furthermore antibiotic be difference because could make with different company. Also on resistant result is not on a good interpretation because if I compare my results with my control some of them show the resistance but if I compare with zone chart provided is susceptible or intermediate I would suggest that in order to improve my experiment results then I should undergo once again until I produce the results that is more accurate and valid. It has been suggested that the results of antimicrobial susceptibility test can affect both the clinician’s choice of antimicrobial therapy and the patient’s outcome. (Daniel J. Diekema 2003)
CONCLUSION
To improve the experiment more study is necessary to correct the mistakes in the investigation, thus to have good results. However generally the experiment went good mainly in haemolytic test where Streptococci pyogens show clear zone of haemolysis when clear the colour of the agar as they breakdown erythrocytes as we expected.
REFERENCE
Hejazi, A. And Falkiner., F.R. 1997. Serratia marcescens. J. Med. Microbiol (online), pp903-912. Accessible at: http://jmm.sgmjournals.org/content/46/11/903.long (Accessed: 18/11/2013)
Dhaka, B.K., Bower, J.M.and Mulvey, M.A.2009. Pili-Fimbriae. Encyclopaedia of Microbiology(online), pp470. Accessible at: http://www.sciencedirect.com/science/article/pii/B9780123739445000511 (Accessed: 18/11/2013)
O’Brien, D. K. and Melville, S.B. 2004. Effects of Clostridium perfringens Alpha-Toxin (PLC) and Perfringolysin O (PFO) on Cytotoxicity to Macrophages, on Escape from the Phagosomes of Macrophages, and on Persistence of C. perfringens in Host Tissues. American Society for Microbiology (online), pp2204-5206. Accessible at: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC517428/ (Accesses : 18/11/2013)
Tan, T. Y., C. McNulty, A. Charlett, N. Nessa, C. Kelly, and T. Beswick.2003. Laboratory antibiotic susceptibility reporting and antibiotic prescribing in general practice.J. Antimicrob. Chemother.51:379-384.9. Accessible at http://www.ncbi.nlm.nih.gov/pmc/articles/PMC404617/ (Accessed: 20/11/2013)
Schentag, J. J., C. H. Ballow, A. L. Fritz, J. A. Paladino, J. D. Williams, T. J. Cumbo, R. V. Ali, V. A. Galletta, M. B. Gutfeld, and M. H. Adelman.1993. Changes in antimicrobial agent usage resulting from interactions among clinical pharmacy, the infectious disease division, and the microbiology laboratory.Diagn. Microbiol. Infect. Dis.16:255-263. Accessible at http://www.ncbi.nlm.nih.gov/pmc/articles/PMC404617/ (Accessed; 20/11/2013)
Johannes F. Van Den Bosch,1Ursula Verboom-Sohmer,1Peter Postma,1Johannes De Graaff,2andDavid M. MacLaren1 1980Mannose-sensitive and mannose-resistant adherence to human uroepithelial cells and urinary virulence of Escherichia coli.Accessible at http://www.ncbi.nlm.nih.gov/pubmed/?term=Verboom-Sohmer%20U%5Bauth%5D
British Society For Antimicrobial Chemotherapy ,Susceptibility Testing 2013 at: http://bsac.org.uk/wp-content/uploads/2012/02/Version-12-Apr- (Accessed 17/11/2013)
Websites
www1
http://www.lecithinguide.info/lecithin-is-powerful.html. (Accessed 18/11/2013)
www2-
http://www.ncbi.nlm.nih.gov/books/NBK7611/ (Accessed :19/11/2013)
www3
2013_final.pdfhttp://www.vetbact.org/vetbact/?biochemtest=1&PHPSESSID=00b630304e2e57b12bc34a9a8bb7497b#id27 (Accessed 23/11/2013)
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2180968/pdf/je14651169.pdf (Accessed 18/11/2013)
APPENDIX
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