the effect of age on isolated bacteria from cancer patients
TRANSCRIPT
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Mashooma Khalifa
BIO 415N
Spring 2011
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Recently in the Indian Journal of Cancer the
article, The effect of age on the bacteria
isolated and the antibiotic-sensitivity pattern in
infections among cancerpatients was published
in December of 2010.
Focus of this article is how age can have an
effect on the types of bacteria present in cancer
patients as well as its antibiotic treatment
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As one ages, the cells of the individual start to loseits ability and the individual begins to respond less
frequently to stress while an increase in homeostatic
imbalances occurs.
The body becomes susceptible to cancer as well as
to vast amounts of infectious diseases
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As one ages, do the types of bacteria differ from agegroups?
Does the effect of antibiotics increase or decrease as
one ages, especially if diagnosed with cancer?
Can the effectiveness of the antibiotics be
controlled?
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*Patients with cancer are predisposed to infections,and on many occasions, no focus of infection can be
established
*Pathologists enforce the treatment ofempirical
antibiotic therapy in these patients
*This therapy involves prescribing certain
medications even though an accurate diagnosis
cannot be made [trial & error]
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*It can be hypothesized, that the effectiveness of the
antibiotics may change over time & as age progresses,
which would result in changes in the antibiotic
sensitivity (Kumar et all. 2010).
*That is why, the main purpose of this experiment is to
test such a hypothesis on seriously ill patients, such ascancer patients
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*Laboratory experiments were conducted at Tata Memorial
Hospital in Mumbai, India during the fall of 2010.
*The experiment had to be conducted secretly so that the
neither the patients nor their family & friends be affected in
any physical, emotional or mental way
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Bacterial samples were selected from patients with a
constant history of infections, were selected for this
experiment
A total of 935 isolates were collected of patients
admitted under the Medical Oncology service
Antibiotic sensitivity & microbiological profile of
infections were divided according to three age
groups: 12-19 years, 20-49 years & >50 years.
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Five pathogens were isolated/used:
Pseudomonas species
Enterococcus species
Staphylococcus aureus
Escheria coli
Klebsiella species
Figure 1 Figure 2
Figure 3 Figure 4
Figure 5
http://upload.wikimedia.org/wikipedia/commons/8/85/Pseudomonas_aeruginosa_01.jpghttp://en.wikipedia.org/wiki/File:Enterococcus_histological_pneumonia_01.pnghttp://en.wikipedia.org/wiki/File:Staphylococcus_aureus_VISA_2.jpghttp://en.wikipedia.org/wiki/File:EscherichiaColi_NIAID.jpghttp://en.wikipedia.org/wiki/File:EscherichiaColi_NIAID.jpghttp://en.wikipedia.org/wiki/File:Staphylococcus_aureus_VISA_2.jpghttp://en.wikipedia.org/wiki/File:Enterococcus_histological_pneumonia_01.pnghttp://upload.wikimedia.org/wikipedia/commons/8/85/Pseudomonas_aeruginosa_01.jpg -
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The pathogens were easily identifiable via the
Analytical Profile Index or API test
It is a rapid biochemical test system that
allows to identify a limited number of Gram-
Negative Enterobacteriacea
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All test chambers are rehydrated by inoculation with a saline
suspension of a pure culture of the bacterial strain
After incubation in a humidity chamber for 18-24 hours at 37C,the color reactions are read. The results of the test reactions are
converted to a seven-digit code. The code can then be looked upin the database book or software
Common substrates tested on the API is:
CITCitrate for the reaction: citrate utilization UREUrea for the reaction: urea hydrolysis
INDTryptophan for the reaction: indole production
SACSucrose for the reaction: fermentation/oxidation
OXOxidase for the reaction: Oxidation
H2SNa Thiosulfate for the reaction: H2S Production
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Inoculation of chambers with the
saline bacterial culture
Code is interpreted by only writing the
+ reactions that occurred, for
example in set of boxes, two out of
four reactions were positivethus the#2 is written in the box below
Sample of the API Test, the roman numerals on top indicate the 7 set of
boxes in which 7 digits will be recorded
Figure 6
Figure 7
Figure 8
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The antibiotic-sensitivity pattern wasidentifiable via the Kirby-Bauer AntimicrobialSusceptibility Test or the KB Test
This test was done following standardprotocol, such as the CLSI standard (Clinical
& Laboratory Standards Institute)
The samples obtained were then mapped onspecial microbiological software
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An agar plate is inoculated with the bacterial culture
Filter paper is impregnated with an antibiotic that is
then placed on the plate using an Automatic Disk
Dispenser
The antibiotic would diffuse from the disk onto the
agar plate
Based on the characteristics of the bacteria, around
each disk there will be a zone of inhibition
(diameter) that is measured to determine the
resistance and susceptibility values
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Example:
The antibiotic Penicillin Staphylococci in the presence of
Penicillin, has a resistance value of 28mm and a
susceptible value of 29mm
For a pathogen to be resistant to Penicillin, it must have a
diameter of < 28mm
Sample of a KB Test, in which disks of Erthromycin [E-2],
Novobiocin [NA-30], Penicillin [P-10], Polymyxin B [PB-
300], Vancomycin [VA-30] and Streptomycin [S-10] were
impregnated on a bacterial culture ofEscheria coli.
Figure 9
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The results determined that there was no significant
change in the distribution of Gram-Positive & Gram-
Negative bacteria with age
HOWEVER,
There was an increase in the occurrence of
Enterobacteriacea group with increasing age
There was a decrease in infections caused by
nonlactose fermenters with increasing age
Most antibiotics decreased in activity with increasing
age
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Gram-Negative bacteria were the predominant
group of bacteria66.96% of the isolates
An increase in infections caused byE.coli &
Klebsiella with increasing age was found
There was a decrease in infections caused by the
Pseudomonas species with increasing age
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Site of Infections:51.76% at the blood stream
15.08% at the respiratory tract
12.62% at the GI tract
11.55% at the skin & soft tissue
3.32% at the urinary tract
5.67% at other sitesTable 1 Table 2
Percentage of occurrence of bacterial pathogens
among the three age groups
Organism:% by age group (years)
12 19 20 49 > 50
Pseudomonas sp (245) 30.01 26.01 23.87Enterococcus sp (109) 12.72 11.75 10.70S. aureus sp (107) 12.14 10.40 13.17E. coli (106) 8.67 11.37 13.17Klebsiella sp (99) 7.51 11.37 11.11
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Extended-spectrum -lactamase (ESBL) are enzymes whichhydrolyze extended-spectrum cephalosporin's with anoxyimino side chainESBLs are enzymes made by some germs. Germs can cause you to
get sick and antibiotics are used to treat or kill these germs. When
germs have ESBLs, many antibiotics do not work.
ESBL production increased from 10.52% in patients 12-19years to 19.31% in patients 20-49 years to 24.88% in patients50 years or older
The production of ESBL was statistically significant wheneach age group was compared with each other resulting witha P value of < 0.01
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The most active antibiotic agent against Gram-Positive
bacteria was Linezolid with no resistance, but it wasuniversally sensitive to the Enterococcal species
Vancomycin resistance (83%) was common among
Enterococcal speciesbut there was a decrease infrequency as age increased
Vancomycin, Linezolid and Teicoplanin were most active
again Staphylococcal species with no resistance, irrelevant
with ageDecreasing trend of activity for meropenam was
statistically significant against Pseudomonas species (73.3-
41.2%) and of aminoglycosides forAcinetobacterspecies
(61.1-17.4%) as age increased
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An evident trend of decreasing antibiotic
resistance against pathogens with increasing age
The reasons could be greater exposure to
antibiotics during life, higher ESBL production
and the need for more invasive procedures than
Empirical therapy
Only improving efficacy of polymyxin &colistin again Pseudomonas species (40-91.7%)
was observed
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Article
Kumar P, Medhekar A, Ghadyalpatil N.S, Noronha V, Biswas S, Kurkure P, Nair R, Kelkar R, Banavali S.D. 2010. The
effect of age on the bacteria isolated and the antibiotic-sensitivity pattern in infections among cancer patients. Indian
Journal of Cancer [Internet] [cited 2011 Mar 14]; 47(4):391-396. Available from:
http://corvette.salemstate.edu:2561/ehost/detail?hid=8&sid=cfba2cdc-a31e-4f75-baad-
5d212b89487f%40sessionmgr10&vid=3&bdata=JkF1dGhUeXBlPWNvb2tpZSxpcCxjcGlkJmN1c3RpZD1zc2Mmc2l0Z
T1laG9zdC1saXZlJnNjb3BlPXNpdGU%3d#db=bxh&AN=BACD201100046666
Images
Figure 1: http://phil.cdc.gov/PHIL_Images/20050113/c6f15fe678d24b739134735e2296e3f7/6687_lores.jpg
Figure 2: http://phil.cdc.gov/phil_images/20030114/16/PHIL_2899_lores.jpg
Figure 3: http://phil.cdc.gov/PHIL_Images/11157/11157_lores.jpg
Figure 4: http://www.niaid.nih.gov/SiteCollectionImages/topics/biodefenserelated/e_coli.jpg
Figure 5: http://www.flickr.com/photos/ajc1/3787968951/ Figure 6: http://www.biotech.ug.edu.pl/odl/biochem/picbiochem/api2.jpg
Figure 7: http://www.biotech.ug.edu.pl/odl/biochem/picbiochem/api4.jpg
Figure 8: http://www.biotech.ug.edu.pl/odl/biochem/picbiochem/apiE20.JPG
Figure 9: Microbiology Department. Salem State University. 11/3/2010.
Tables
Table 1: http://www.indianjcancer.com/articles/2010/47/4/images/IndianJournalofCancer_2010_47_4_391_73574_b1.jpg
Table 2: http://www.indianjcancer.com/text.asp?2010/47/4/391/73574