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Bacteriological analysis of the water in El Caño Martín Peña
Detection and quantification of Enteric Pathogenic Bacteria Salmonella and Shigella spp.
1 / march/ 2013
Mario R. BarreraGeorge M. RiveraPaola TabaroMentor: Prof. Mayra RolónUniversity of the Sacred Heart
Meet the Team
Mario R.Barrera
PaolaTabaro
George M. Rivera
Mentor:Prof: Mayra Rolón
Meet the Team
2012
2013
Caño Martín Peña
Is an invaluable natural habitat for the Metropolitan area of San Juan.
Caño Martín Peña
+ During the recurring years the mangroves and lagoons stretched of the Caño have been contaminated and stretched out into smaller measurements.
Caño Martín PeñaThe deterioration of this area has taken its toll on the residents lifestyles.
How to Improve !!
The decontamination of the lagoon that boards the community.
Rehabilitation of the estuary system.
To broaden the community’s knowledge of the microbial presence and the harms that can be encountered.
Our objectives
+ To detect the presence of pathogenic enteric bacteria such as Salmonella and Shigella SPP.
+ To acquire knowledge of the microbial presence found in the Caño Martín Peña.
+ Refine research skills through different bacteriological techniques.
+ Analyze and compare the degree of contamination found with past investigations.
+ Raise awareness to the community.
Hypothesis
In the past year our fellow classmates were able to detect the presence of Shigella serotypes A,B and C.
The Caño Martín Peña has excessive levels of coliforms, indicators of fecal matter that represent a hazard for the community.
Hypothesis
+ Because of this information we can say there is a high probability of encountering Salmonella and Shigella SPP. Bacteria due to the high coliform levels resulting from water contamination by fecal matter.
Coliform bacteria
Rod-shaped Gram negative bacillus.
Lactose fermenters with the production of acid and gas when incubated at 35-37°C.
Easy to culture and their presence is used to indicate fecal contamination by other pathogenic organisms in the water.
Enterobacteriaceae
Total coliforms: Escherichia Klebsiella Serratia Citrobacter
Fecal coliform: Eschericia coli
Escherichia coli
Klebsiella pneumoniae
Enterobacter
Serratia marcescens
Citrobacter
Salmonella spp. Vs Shigella spp.
Salmonella spp.
Rod-shaped Gram negative bacillus
Non-spore forming. Motile enterobacteria
Facultative anaerobes
Hydrogen sulfide production which can be detected on a TSI growth media
Shigella spp.
Rod-shaped Gram negative bacteria
Non-spore forming and non-motile bacteria
Salmonella Pathogenicity
Most of the infections are due to the ingestion of contaminated food.
Salmonella is responsible for various illnesses such as: Typhoid fever Paratyphoid fever Foodborne illnesses
Enteritis Salmonella also known as food poisoning Salmonella.
Shigella Pathogenicity
Most of the infections are caused via ingestion.
The illness is known as Shigellosis.
During infection, Shigella typically causes dysentery .
Invasion of Shigella Bacterial multiplication Spreading to epithelial
cells Tissue destruction
Investigation Site
Station #1
Width : 8 Ft.
Depth: 2.05 Ft
Observations Boat nearby Carton Boxes
Station #2
Width: 3 – 5 Ft
Depth: 4 Ft
Observations Current of water
coming from a pipe. Fishing traps
Station #3
Width: 8 Ft.
Depth: 2 Ft.
Observations Samples were taken
at the shore, because difficult access.
Garbage bags
Methodologies
Dilution of sample
Membrane filtration
Cultivation SS Agar MacConkey Agar EMB RVS
Identification Enterotubes
Procedure
DilutionMembrane Filtration
Cultivation
Dilution of sample
At the Laboratory
Membrane filtration 0.45 μm
At the Laboratory
Membrane filtration
Pure sampleDiluted Sample
1:1000
Cultivation
At 37º C.
24 – 48 Hours
Selective Mediums
Salmonella Shigella Agar
MacConkey Agar
Selective Mediums
RVS EMB
Taxonomic Determination: Enterotube System
Taxonomic Determination: Enterotube System
Enterotube System
Citrobacter freundii
Serratia plymuthica
Sampling Dates
M1 – 30/ October/ 2012 M2 – 28/ November/ 2012 M3 – 7/ December/ 2012 M4 – 30/ January/ 2013 M5 – 11/ February/ 2013
Types of Bacteria
Characteristics of the Colony
Bacteria
Salmon Colony Klebsiella ozanae
Pink Colony Enterobacter cloacae
Pink Colony Klebsiella pneumoniae
Pink colony Enterobacter aerogenes
Transparent Colony Shigella
Transparent Colony Yersinia
Black Colony Citrobacter freundi
Pale pink Serratia plymuthica
Metallic Green E. coli
Types of Bacteria
Bacteria
Serratia marcenscens
Enterobacter aglomerans
Enterotubes
Shigella Enterotube
Findings
Year Quantity
2008 118,000
2009 3,335,000
2010 6,800,000
2011 60,000,000
2012 69,000,000
2013 25,333,333
Year Quantity
2000 4,000,000
2001 5,500,000
2002 4,000,000
2003 2,000,000
2004 100,000
2005 73,000
2006 80,000
2007 57,000
Number of bacteria/ ml of water
Findings on Dilutions
Station #1
1 col in 10¯⁷ : 10,000,000 bact/ml
Station #2 4 col in 10⁻⁶: 4,000,000 bact/ml
Station #3 62 col in 10⁻⁶: 62,000,000 bact/ml
Findings on Filtration
Station#1 Pure > 100 col (TNTC) Dilute: 5 col * 1000: 5,000 bact/100ml
Station #2 Pure>100 col (TNTC) Dilute: 40 col * 1000: 40,000 bact/100ml
Station #3 Pure>100 col (TNTC) Dilute: 3 col * 1000: 3,000 bact/100ml
Citrobacter freundiCedecea lapageiKlebsiella pneumoniaeYersinia enterocoliticaCitrobacter freundiCedecea lapageiEnterobacter agglomeransKlebsiella ozaenae
Enterobacter agglomerans
Conclusions
The high level of contamination is due to:
Stagnant water in some areas.
Sewage and water discharge in to the cano.
Deposition of debris into the community.
Conclusions of the Research
From 2004 to 2008 the amount of bacteria were less than a million/ ml of water.
This could be due to the removal of debris
Effort of government and people that lives in the community.
+ During our Research period we noticed a sudden decrease in the level of bacteria.
+ Although we were not able to isolate colonies of Salmonella, we did isolate Citrobacter freundi, Yersinia enterocolitica and Shigella Spp.
+ Which are indicative of the severity of contamination and a possible source of contamination with pathogens to the residents of nearby areas.
We recommend:
To maintain the movement of dredging and cleaning of the entire San Juan Estuary.
We saw that according to data, levels of bacteria were down under a million. With a little of effort is possible to maintain to provide a better life to the people.
To Help organizations and people that live there in a constant way not only, to show interest when elections are near.
References
Http://www.mayoclinic.com/health/shigella/DSOO719
Http://www.cdc.gov/salmonella/enteritidis/
Centers for Disease Control and Prevention. October 2005. Shigellosis. http://www.cdc.gov/nczved/divisions/dfbmd/diseases/shigellosis/
Indian and Northern Affairs Canada. May 2003. First Nations Water Management Strategy. Http://www.ainc-inac.gc.ca/ai/arp/es/0506/fnwms/fnwms-eng.asp
References
Public Health Agency of Canada:Notifiable Diseases on-line. December 2003. Shigellosis. http://dsol-smed.hc-sc.gc.ca/dsol-smed/ndis/diseases/shig_e.html
www.safewater.com
World Health Organization. 2005. Guidelines for the control of shigellosis, including epidemics due to shigella dysenteriae type 1. http://whqlibdoc.who.int/publications/2005/9241592330.pdf
United States Pathogenic Microorganisms and Natural Toxins Handbook. January 2002. Shigella. Http://pdf.usaid.gov/pdf_docs/ PNADO152.pdf