Ground water contamination, Amity University

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Microbial contamination of ground water and microbial risks analysis

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Microbial contamination of unlined water through leaching of sewage waste from heavily loaded drains of Delhi, and associated microbial risks analysis:

Microbial contamination of unlined water through leaching of sewage waste from heavily loaded drains of Delhi, and associated microbial risks analysis Presented by: Ashwani Kumar Supervisor Dr. Tanu Jindal* Amity Institute of Environment Toxicology Safety and Management AMITY UNIVERSITY (UTTAR PRADESH), (tjindal@amity.edu)

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Water POLLUTION Water Pollution – Degradation of water quality making it unsuitable for its intended use. Types of pollutants can be biological pathogens, industrial chemicals, heavy metals, fertilizers , sediment, heat, radioactive isotopes, fecal Coliform bacteria, sewage, nutrients or organic materials. Impacts of Water Pollution: ANOXIA: Low level O2 in the bottom surface of water. BOD: A chemical procedure for determining how fast biological organisms use up oxygen in a body of water. It is usually performed over a 5-day period at 20° Celsius. FLOCCULATION: Aggregation of solids to hindered the flow of water. AESTHETIC IMPACTS: Undesirable tastes or odors. TOXINS: Toxic substances released by various biological pollutants. EUTROPHICATION: Process where water bodies receive excess nutrients that stimulate excessive plant growth. BIOMAGNIFICATIONS: WATER BORN DISEASE: Fecal Coliform Bacteria: Population of fecal coliform bacteria in water makes it unsafe for drinking.

Total coliform group, cont. :

Total coliform group, cont. Citrobacter Use citrate as a carbon source Found everywhere including in soil, water, wastewater, human intestine Rarely source off illness (except urinary tract & infant meningitis) Enterobacter Highly motile Part of normal flora of human intestinal tract Several strains can be pathogenic and cause urinary tract infection Escherichia Inhabitants of the gastrointestinal tracts of warm-blooded animals Provide a portion of the microbial–derived vitamin K E.coli – most numerous from this group Klebsiella Widely distributed in nature Occurring in soil, water, grain, vegetation, wood pulp, other… Opportunistic pathogens Can cause pneumonia, urinary tract infections, other infections Associated with coliform re-growth in large water distribution systems Klebsiella pneumoniae ( from feces) – 60 – 80 % shows as positive in FC tests Others Budvicia, Erwinia, Leclercia, Serratia Found only in the environment (no fecal origin)

General Characters of coliform bacteria:

General Characters of coliform bacteria Gram-negative bacilli or Coccobacilli. Non-spore forming. Mobile with peritrichate flagella. Some are non-motile e.g. Klebsiella and Shigella . Non-capsulate except Klebsiella species and Salmonella typhi . Aerobe and facultative anaerobe . Can grow on ordinary media. Selective and differential media e.g. MacConkey’s agar. Oxidase negative, Catalase positive, reduce nitrate to nitrite.

Classification of Enterobacteriaceae Family:

Classification of Enterobacteriaceae Family According to their Cultural Characters: (Growth on MacConkey’s agar medium) Lactose – fermenters (rose pink colonies) include Coliform group: Escherichia , Klebsiella , Citrobacter and Enterobacter . Non-lactose fermenters (pale yellow colonies) include: Salmonella, Shigella (except Shigella sonnei ), Yersinia , and proteus species. Late lactose fermenters (after 48 hours incubation) include: Shigella sonnei . According to their pathogenicity : Primary pathogens: Salmonella , Shigella , Yersinia and certain strains of E. coli . Opportunistic pathogens: Coliform bacilli .

Virulence Factors Associated with Enterobacteriaceae:

Virulence Factors Associated with Enterobacteriaceae Endotoxin. Capsular polysaccharide. Antigenic H phase variation of Salmonella species. Adhesion factors (Fimbriae or pili). Intracellular survival and multiplication. Polysaccharide chain in cell wall resists serum killing by complement. Inhibition of unrelated organisms by enterocines production. Resistance to antimicrobial agents. Production of iron-chelating siderophores that liberate iron essential for growth of bacteria from transferrin or lactoferrin .

Genus Escherichia:

Genus Escherichia  It contains five species; the most important is Escherichia coli (E. coli) Diseases caused by E. coli (A) Diarrheogenic E. coli: Six groups of E. coli cause diarrhea. Each associated with: Certain serotype Has specific virulence factors. Manifest distinct disease pathology. (B) Non Diarrheogenic E. coli .

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Diarrheogenic E. Coli group Site Serotype Disease Virulence factors Pathogenesis identification Enterotoxigenic E. Coli (ETEC) Small intestine 06,08 Travelers’ diarrhea, diarrhea in malnourished infant Colonization factor antigens (CFAs) encoded on plasmid. two toxins: LT (heat labile) ST (heat stable) enterotoxins encoded on plasmid Hypersecretion of fluids and electrolytes into gut lumen  watery diarrhea. LT  ligated ileal loop, steroid stimulation in adrenal tumor cells, EIA ST  diarrhea in mice, DNA probe Enteropathogenic E. Coli (EPEC) Small intestine 018 Infant diarrhea in developing countries Bundle forming pili  localized adherence Destruction of microvilli  malabsorption, watery diarrhea (mucoid, no blood). Serotyping with specific antisera. Enteroinvasive E. Coli (EIEC) Large intestine 029 Diarrhea in children (like bacillary dysentery) Invasion of epithelial cell of colon Intracellular multiplication. Extension to adjacent epithelial cells  watery diarrhea (blood, mucus, pus in stool). Serney’s Test. Enterohaemorrhagic E. Coli (EHEC) or Verotoxigenic E. coli (VTEC) Large intestine 0157 Hemorrhagic colitis, hemolytic uremic syndrome (HUS) Verotoxin (cytotoxic on vero cell line tissue culture) Inhibition of protein synthesis, destruction of intestinal microvilli  decrease absorption of water and electrolytes (like shiga toxin)  bloody diarrhea. Serotyping with specific antisera. No sorbitol fermentation. Vero cell tissue culture. Enteroagreggative E. coli (EAEC) Small intestine Infant diarrhea Bundle forming pili. heat stable enteroaggregative enterotoxin. Adherence to intestinal mucosa  watery, mucoid diarrhea. adherence to HEP-2 tissue culture. DNA probe. Diffusely adherent E. coli (DAEC) Small intestine Infant diarrhea Characterized by adherence to cultured cells Stimulate elongation of intestinal microvilli with bacteria embedded in the cell membrane  watery diarrhea. DNA probe.

Biochemical reactions of coliform bacilli and proteus vulgaris:

Biochemical reactions of coliform bacilli and proteus vulgaris E. coli Klebsiella species Citrobacter species Proteus vulgaris Motility + - + + Capsule  + - - Swarming growth - - - + Blood haemolysis  - - - Glucose Fermentation + + + + Lactose fermentation + + + - Maltose fermentation + + + + Mannite fermentation + + + - Sucrose fermentation + + + +

Biochemical reactions of coliform bacilli and proteus vulgaris:

Biochemical reactions of coliform bacilli and proteus vulgaris E. coli Klebsiella species Citrobacter species Proteus vulgaris Indole production + - - + Methyl red reaction + - + + Voges proskauer - + - - Citrate utilization - + + - Urease production - + - + H 2 S production - - + + Gelatin liquefaction - - - + + Sugars are fermented with acid and gas production.  Some strains give positive while others give negative results.

Okhala drain Delhi Sample:

Okhala drain Delhi Sample

Isolation of single colony:

Isolation of single colony

Growth of colony in NA and EME Agar plate:

Growth of colony in NA and EME Agar plate

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THANK YOU VERY MUCH FOR YOUR KIND ATTENSION Ashwani Kumar bioashwani@gmail.com

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