Bacteriological analysis of the water in the Caño Martín Peña

Bacteriological analysis of the water in the Caño Martín Peña

Bacteriological analysis of the water in El Cao Martn Pea Detection and quantification of Enteric Pathogenic Bacteria Salmonella and Shigella spp. Mario R. Barrera George M. Rivera Paola Tabaro Mentor: Prof. Mayra Roln University of the

Sacred Heart 1 / march/ 2013 Meet the Team George M. Rivera Mario R. Barrera Mentor: Prof: Mayra

Roln Paola Tabaro Meet the Team 2 0 1 2

2 0 1 3 Cao Martn Pea Is an invaluable natural habitat for the Metropolitan area of San Juan.

Cao Martn Pea + During the recurring years the mangroves and lagoons stretched of the Cao have been contaminated and stretched out into smaller measurements. Cao Martn Pea

The 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 communitys 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

Cao Martn Pea. + 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 Cao Martn Pea 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 3537C. 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 Dilution of sample At the Laboratory Membrane filtration 0.45

mm At the Laboratory Membrane filtration Pure sample Diluted Sample 1:1000 Cultivation At 37 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 Year Quantity 2000

4,000,000 2008 118,000 2001 5,500,000 2009

3,335,000 2002 4,000,000 2010 6,800,000 2003

2,000,000 2011 60,000,000 2004 100,000 2012

69,000,000 2005 73,000 2013 25,333,333 2006

80,000 2007 57,000 Number of bacteria/ ml of water Findings on Dilutions Station #1

1 col in 10 : 10,000,000 bact/ml Station #2 4 col in 10: 4,000,000 bact/ml Station #3

62 col in 10: 62,000,000 bact/ml Findings on Filtration Citrobacter freundi Pure > 100 col (TNTC) Cedecea lapagei Dilute: 5 col * 1000: 5,000 bact/100ml Klebsiella pneumoniae Yersinia enterocolitica

Station #2 Citrobacter freundi Pure>100 col (TNTC) Cedecea lapagei Dilute: 40 col * 1000: 40,000 bact/100ml Enterobacter agglomerans Klebsiella ozaenae Station #3 Pure>100 col (TNTC) Enterobacter Dilute: 3 col * 1000: 3,000 bact/100ml

agglomerans Station#1 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/DSOO 719 Http://www.cdc.gov/salmonella/enteritidis/ Centers for Disease Control and Prevention. October 2005. Shigellosis. http://www.cdc.gov/nczved/divisions/dfbmd/diseas es/shigellosis/ Indian and Northern Affairs Canada. May 2003.

First Nations Water Management Strategy. Http://www.ainc-inac.gc.ca/ai/arp/es/0506/fnwms/f nwms-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/shi g_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

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