"1 Progression of an infection to bacteraemia in immuno-suppressedmiceStudent Name:Student Number: Progression of an infection to bacteraemia inimmuno-suppressed mice1 2 Progression of an infection to bacteraemia in immuno-suppressedmiceStudent Name:Student Number:Part AIntroductionInjuries received due to burning have more long-lasting effects than previouslythought. Extensive burning potentially increases the risk of development of veryserious infections. These infections are extremely deadly, as they are potentiallyfatal. Burn injuries result in skin damage, which in turn, result in an entry ofunnecessary and dangerous bacterial species in the blood. This condition istermed as bacteraemia, and its development is the phenomenon that ultimatelyleads to life-threatening repercussions. Bacterial species that have been known toaffect and contaminated burned injury sites include Pseudomonas aeruginosa,which is among the most dangerous bacterial species. Their tendency to invadeother tissues of burn victims in addition to their resistance to antibiotics makethem extremely formidable. Studies attempting to solve the riddle of itsprogression and determining its mode of action have been conducted. Relevanthypotheses and ideas regarding its progression methods are tested for theirvalidity in this study. Main experiments include testing on mice to check howbacterial species progress from a simple local skin infection to bloodcontamination in immune-suppressed patients. Herein, this study is about thedetermination of progress of specific dangerous bacterial species from infection tothe potentially fatal bacteraemial stage. 2 3 Progression of an infection to bacteraemia in immuno-suppressedmiceStudent Name:Student Number:MethodsMain information has been gathered using testing and experiments done on tenselected mice. These mice proved to be the main samples to be used in thesestudies. They were selected based on their reaction towards external stimuli. Onceselected, it is appropriate and necessary to divide them into selected groups. The division into selected groups is done to classify them based on control and1 test . In order to gain relevant and accurate data, the best way is to include fivemice in each of these groups. An equal number of members in both groups alsofavor accurate comparison during times of testing and generation of results. Thecontrol group is mainly used for gaining data of mice with normal functions.Mice classified into the control group had normal functions of their immunesystems. Any reactions and results generated from this group are to be taken asthe general reaction of mice to any kind of testing or experimentation. Deviationsfrom these reactions are to be taken as abnormalities, and the extent of these2 abnormalities are observed, calculated and incorporated into the results . 1 Dantas RC, Ferreira ML, Gontijo-Filho PP, Ribas RM. Pseudomonas aeruginosa bacteraemia: independentrisk factors for mortality and impact of resistance on outcome. Journal of medical microbiology. 2014 Dec1;63(12):1679-87.2Hodgson LE, Dragolea N, Venn R, Dimitrov BD, Forni LG. An external validation study of a clinicalprediction rule for medical patients with suspected bacteraemia. Emergency Medicine Journal. 2015 Aug5:emermed-2015. 3 4 Progression of an infection to bacteraemia in immuno-suppressedmiceStudent Name:Student Number:The test group of mice, as is obvious, is treated with certain chemicals. Treatmentof members of this group with specific doses of cyclophosphamide is the mainhallmark of the conducted experiments. Main dose given on Day 0 is preferably200 mg/kg. The dosage is given for the sole purpose of suppressing theirimmunity systems, which in turn, will help in generating relevant results.One of the main processes that happen in most living organisms in response to3 any kind of infections is a drastic increase in the production rate of leukocytes .Leukocytes are also commonly known as white blood cells. They are the main‘soldiers’ produced by bodies in order to cope with any kind of external invasion.This includes invasion by bacterial or any other microbial species. This protectionextends to foreign particles of any nature and it is extremely effective in dailyprotection of most multi-cellular beings. In order to gain valuable data pertainingto the needs of this study, it is necessary that leukocyte actions be suppressed.This task is performed efficiently by the aforementioned cyclophosphamide.Another important activity performed on Day 0 is infection of bacteriadeliberately by injecting them into the upper skin layer of mice of control and testgroups. Injecting them into the upper skin layer gives a very accurate3Milstone AM, Elward A, Song X, Zerr DM, Orscheln R, Speck K, Obeng D, Reich NG, Coffin SE, PerlTM, Pediatric SCRUB Trial Study Group. Daily chlorhexidine bathing to reduce bacteraemia in critically illchildren: a multicentre, cluster-randomised, crossover trial. The Lancet. 2013 Apr 5;381(9872):1099-106. 4 5 Progression of an infection to bacteraemia in immuno-suppressedmiceStudent Name:Student Number:representation of how bacterial species may invade in the case of patientssuffering from burns, as their upper skin layers are damaged as well.Upon completing injection, each mouse has been weighed daily for a period ofnine days. Weighing a mouse gives a very good indication of overall state ofhealth of an animal. It is extremely important to check the well-being of testedmice daily, as progression of bacteria directly corresponds to gradual worseningof health. Taking samples of blood from each mouse also gives very accurate4 information regarding their body conditions .Taking blood samples gives best results when it is done daily. Blood samplingand testing has been done for a period of six days following original injection of5 bacteria . Analysis of procured data from these studies is to be tabulated in Excel.A table is dedicated to giving information regarding daily body weight in gramsover the period of nine days following infection. A similar table detailing4Munita JM, Murray BE, Arias CA. Daptomycin for the treatment of bacteraemia due to vancomycin- resistant enterococci. International journal of antimicrobial agents. 2014 Nov 30;44(5):387-95. 5Stentzel S, Sundaramoorthy N, Michalik S, Nordengrün M, Schulz S, Kolata J, Kloppot P, Engelmann S,Steil L, Hecker M, Schmidt F. Specific serum IgG at diagnosis of Staphylococcus aureus bloodstreaminvasion is correlated with disease progression. Journal of proteomics. 2015 Oct 14;128:1-7.5 6 Progression of an infection to bacteraemia in immuno-suppressedmiceStudent Name:Student Number:leukocyte count of six days following infection is also necessary. Observationsand deviations of data are to be recorded for coming to a conclusion.Average 30.00 25.00 20.00 15.00 Average 10.00 5.00 0.00 Day 1 Day 2 Day 3 Day 4 Day 5 Day 6 Day 7 Day 8 Day 9 6 7 Progression of an infection to bacteraemia in immuno-suppressedmiceStudent Name:Student Number:Standard deviation 1.00 0.90 0.80 0.70 0.60 0.50 Series1 0.40 0.30 0.20 0.10 0.00 1 2 3 4 5 6 7 8 9 Figure 1: Average and Standard Deviation of weight of CY infected mice ingrams(Created by Self)Average 26.00 25.50 25.00 24.50 24.00 23.50 Series1 23.00 22.50 22.00 21.50 1 2 3 4 5 6 7 8 9 7 8 Progression of an infection to bacteraemia in immuno-suppressedmiceStudent Name:Student Number:Standard Deviation 1.2 1 0.8 0.6 Series1 0.4 0.2 0 1 2 3 4 5 6 7 8 9 Figure 2: Average and Standard Deviation of weight of Control mice ingrams(Created by Self)Average 70 60 50 40 30 Series1 20 10 0 1 2 3 4 5 6 8 9 Progression of an infection to bacteraemia in immuno-suppressedmiceStudent Name:Student Number:Standard Deviation 60 50 40 30 Series1 20 10 0 1 2 3 4 5 6 Figure 2: Average and Standard Deviation of leukocyte count in CY infectedmice(Created by Self)Average 120 100 80 60 Series1 40 20 0 1 2 3 4 5 6 9 10 Progression of an infection to bacteraemia in immuno-suppressedmiceStudent Name:Student Number:Standard Deviation 60 50 40 30 Series1 20 10 0 1 2 3 4 5 6 Figure 2: Average and Standard Deviation of leukocyte count in Controlmice(Created by Self)10 11 Progression of an infection to bacteraemia in immuno-suppressedmiceStudent Name:Student Number:Reference ListDantas RC, Ferreira ML, Gontijo-Filho PP, Ribas RM. Pseudomonas aeruginosabacteraemia: independent risk factors for mortality and impact of resistance onoutcome. Journal of medical microbiology. 2014 Dec 1;63(12):1679-87. Hodgson LE, Dragolea N, Venn R, Dimitrov BD, Forni LG. An externalvalidation study of a clinical prediction rule for medical patients with suspectedbacteraemia. Emergency Medicine Journal. 2015 Aug 5:emermed-2015. Milstone AM, Elward A, Song X, Zerr DM, Orscheln R, Speck K, Obeng D,Reich NG, Coffin SE, Perl TM, Pediatric SCRUB Trial Study Group. Dailychlorhexidine bathing to reduce bacteraemia in critically ill children: amulticentre, cluster-randomised, crossover trial. The Lancet. 2013 Apr5;381(9872):1099-106. Munita JM, Murray BE, Arias CA. Daptomycin for the treatment of bacteraemiadue to vancomycin-resistant enterococci. International journal of antimicrobialagents. 2014 Nov 30;44(5):387-95. Stentzel S, Sundaramoorthy N, Michalik S, Nordengrün M, Schulz S, Kolata J,Kloppot P, Engelmann S, Steil L, Hecker M, Schmidt F. Specific serum IgG atdiagnosis of Staphylococcus aureus bloodstream invasion is correlated withdisease progression. Journal of proteomics. 2015 Oct 14;128:1-7.11 12 Progression of an infection to bacteraemia in immuno-suppressedmiceStudent Name:Student Number:12 "