1 / march/ 2013 Mario R. Barrera George M. Rivera Paola Tabaro Mentor: Prof. Mayra Rolón University of the Sacred Heart

Mario R. Barrera PaolaTabaro George M. Rivera Mentor: Prof: Mayra Rolón

 Is an invaluable natural habitat for the Metropolitan area of San Juan.

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

The deterioration of this area has taken its toll on the residents lifestyles.

 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.

 + 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.

 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.

 Total coliforms:  Escherichia  Klebsiella  Serratia  Citrobacter  Fecal coliform:  Eschericia coli

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

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

 Dilution of sample  Membrane filtration  Cultivation  SS Agar  MacConkey Agar  EMB  RVS  Identification  Enterotubes

Dilution Membrane Filtration Cultivation

Pure sampleDiluted Sample 1:1000

 At 37º C.  24 – 48 Hours

Salmonella Shigella Agar MacConkey Agar

RVSEMB

Citrobacter freundiiSerratia plymuthica

 M1 – 30/ October/ 2012  M2 – 28/ November/ 2012  M3 – 7/ December/ 2012  M4 – 30/ January/ 2013  M5 – 11/ February/ 2013

Characteristics of the ColonyBacteria Salmon ColonyKlebsiella ozanae Pink ColonyEnterobacter cloacae Pink ColonyKlebsiella pneumoniae Pink colonyEnterobacter aerogenes Transparent ColonyShigella Transparent ColonyYersinia Black ColonyCitrobacter freundi Pale pinkSerratia plymuthica Metallic GreenE. coli

Bacteria Serratia marcenscens Enterobacter aglomerans

YearQuantity , ,335, ,800, ,000, ,000, ,333,333 YearQuantity 20004,000, ,500, ,000, ,000, , , , ,000 Number of bacteria/ ml of water

 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

 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 freundi Cedecea lapagei Klebsiella pneumoniae Yersinia enterocolitica Citrobacter freundi Cedecea lapagei Enterobacter agglomerans Klebsiella ozaenae Enterobacter agglomerans

 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.

 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.

 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.

   Centers for Disease Control and Prevention. October Shigellosis. osis/ osis/  Indian and Northern Affairs Canada. May First Nations Water Management Strategy. inac.gc.ca/ai/arp/es/0506/fnwms/fnwms-eng.aspHttp:// inac.gc.ca/ai/arp/es/0506/fnwms/fnwms-eng.asp

 Public Health Agency of Canada:Notifiable Diseases on-line. December Shigellosis. smed/ndis/diseases/shig_e.htmlhttp://dsol-smed.hc-sc.gc.ca/dsol- smed/ndis/diseases/shig_e.html   World Health Organization Guidelines for the control of shigellosis, including epidemics due to shigella dysenteriae type  United States Pathogenic Microorganisms and Natural Toxins Handbook. January Shigella. PNADO152.pdf