Measuring Stream Microbiology: Methods and Preliminary Results Dr. Robert B. Simon Mr. Jonah Stevens Department of Biology SUNY-Geneseo

Problems Measuring Microbial Quality in Streams Which organisms to measure? How to do measurements on the scale required to monitor 7 streams weekly as well as events, especially considering the need for immediate measurement! Samples are held during events - How does this change the results?

The Real Culprits The main route of exposure to illness-causing organisms in recreational beach waters is through direct contact with polluted water while swimming. Bacterial infection (such as cholera, salmonellosis, shigellosis, and gastroenteritis). Viral infection (such as infectious hepatitis, gastroenteritis, and intestinal diseases caused by enteroviruses). Protozoan infections (such as cryptosporidiosis, amoebic dysentery, and giardiasis).

Microbial Quality is Monitored by Measuring Surrogates Coliforms Fecal Coliforms Fecal Streptococci Enterococci Total Heterotrophic Bacteria

Coliforms Found in the gut and feces of warm-blooded animals Aerobic and facultative anaerobic, Gram-negative, non-sport forming rod shaped bacteria that ferment lactose with gas formation within 48 hours at 35 °C. Enterobacteriaceae Membrane Filtration (MF) MF - Dark colony with a metalic sheen on an “Endo” type medium.

Fecal Coliforms A subset of Total Coliforms Fecal coliforms - Produce gas from lactose at 44.5 °C. Prior to its 1986, EPA recommended the use of fecal coliforms as an indicator organism to protect bathers from gastrointestinal illness in recreational waters.

Membrane Filter methods for Total Coliforms and E. coli using Millipore mColiBlue24 Next day results Easy to read with turbid samples Clonal isolates can be saved for further study

Fecal Streptococcus Isolated from the feces of warm-blooded animals A number of species of the genus Streptococcus, such as: S. faecalis, S. faecium, S. avium, S. bovas, S. equinus, S. gallinarum Positive reaction with Lancefield Group D antisera It is not possible to differentiate the source of fecal contamination based on speciation FC/FS ratio can also not be used as a means of differentiating human and animal feces

Enterococcus A subgroup of fecal streptococci, including: S. faecalis, S. faecium, S. gallinarum, S. avium Differentiated by ability to grow in 6.5% NaCl at 10 °C and 45 °C Difco m Enterococcus Agar

Heterotrophic Bacteria Bacto R2A Agar. A low-nutrient medium. Incubation at 25 °C for hours, stimulates the growth of stressed and chlorine-tolerant bacteria Multiple 20  l drops of water dilutions are spotted on plates, and micro-colonies are counted under a dissecting microscope

EPA’s Recommended 1986 Water Quality Criteria for Bacteria in Recreational Fresh Water Based on the epidemiology of water-born disease occurrence as E. coli and Enterococci levels exhibit a strong correlation to swimming-associated gastroenteritis. January 2003 tentative release of new microbial strategy

The End