# Presentation: 122350

## Dale Griffin and Mike Gray

- USGS, St. Petersburg, FL

## The Problem

- The susceptibility of Florida ground waters to recharge-borne microorganisms and pollutants (nutrients, herbicides, pesticides, hydrocarbons, etc.)
- Rising nitrate levels in Florida’s springs
- The need to determine the source of pollutants
-  Agricultural versus residential – waste water, fertilizer, hydrocarbons, pesticides, etc.
- Human versus animal (agricultural and indigenous)

## Enter the Virus

- Bacteriophage

- Animal viruses

- Polio

- Norwalk-like

- Astro

- Astro

-  Many viruses are host specific and 	within a host only infect specific 	types of tissue
-  Host specific viruses can not replicate 	outside of their host – thus when 	they are found in the environment it 	means that its host is impacting that 	environment.
-  Molecular techniques are available 	to detect low concentrations 	of viruses in water (Reverse 	transcriptase – Polymerase chain 	reaction, RT-PCR).

## Reverse Transcriptase - Polymerase Chain Reaction

## Slide 5

## Homosassa Springs

## Mystery Spring 

***Mystery Spring ***

- Average spring vent bacteria count/ml = 5,870
- Average spring vent virus count/ml = 29,900

## Table 1. Microbiology data (spring sample data for 9-25-03)

| Spring | pH/Temperature-oC/EC-μS/TDS-ppm | Indicator Data – Colony Forming Units/100ml (duplicate average) |  |  |  | Virus Polymerase Chain Reaction data (Equivalent volume assayed) |  |  |  |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
|  |  | Enterococci | Clostridium perfringens | Fecal coliforms | E.coli | Enteroviruses | Hepatitis A viruses | Adenoviruses | Cattle enteroviruses |
| Hornsby | 6.9/22.8//298/150 | 0 | 0 | 0.2 | 0.1 | - (0.04L) | - | - | - |
| Poe | 7.4/-/287/143 | 0.1 | 0 | 0 | 0 | - (0.5L) | - | - | - |
| Manatee | 7.3/-/329/164 | 0.2 | 0 | 0.2 | 0.1 | - (0.1L) | - | - | - |

| Spring | pH/Temperature-oC /EC-μS/TDS-ppm | Indicator Data – Colony Forming Units/100ml (duplicate average) |  |  |  | Virus Polymerase Chain Reaction data  |  |  |  |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
|  |  | Enterococci | Clostridium perfringens | Fecal coliforms | E.coli | Enteroviruses | Hepatitis A viruses | Adenoviruses | Cattle enteroviruses |
| Hornsby | 7.1/22.0/271/131 | 0.2 | 0 | 0 | 0 | - (0.3L) | - | - | - |
| Poe | 7.1/22.1/266/133 | 0 | 0 | 0 | 0 | - (0.3L) | - | - | - |
| Manatee | 7.0/22.0/311/155 | 2.0 | 0 | 4.7 | 4.7 | - (0.3L) | - | - | - |

**Table 2. Microbiology data (spring sample data for 12-17-03)**

| Spring | pH/Temperature-oC /EC-μS/TDS-ppm | Indicator Data – Colony Forming Units/100ml (duplicate average) |  |  |  | Virus Polymerase Chain Reaction data  |  |  |  |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
|  |  | Enterococci | Clostridium perfringens | Fecal coliforms | E.coli | Enteroviruses | Hepatitis A viruses | Adenoviruses | Cattle enteroviruses |
| Hornsby | 7.2/22.5/276/139 | 1.2 | 0 | 3.0 | 3.0 | + (0.5L) | - | - | - |
| Poe | 7.2/22.5/273/137 | 0.1 | 0 | 4.5 | 4.5 | ++ (0.5L) | - | - | - |
| Manatee | 7.1/22.5/320/158 | 0.4 | 0 | 14.7 | 14.7 | +++(0.5L) | - | - | - |

- + = light signal via genetic probe/dot blot
- ++ = medium signal via genetic probe/dot blot
- +++ = heavy signal via genetic probe/dot blot

**Table 3. Microbiology data (spring sample data for 4-20-04)**

**SRWMD District rain data – Sept. 03, actual 3.17 (avg. 5.46) – Manatee 2.36**
**                   ****	****               ****Dec. 03, actual 1.43 (avg. 3.17) – Manatee 1.23**
**                                               ****May 04, actual 1.69 (avg. 3.39) – Manatee 3.53**

## Conclusions

- Surface associated waste water and pollutants can impact Florida’s ground water
- Human viruses have been detected in Florida ground water and emanating from spring vents
- Host specific virus assays are a useful tool in determining sources of pollutants
- Agricultural versus residential
- Human versus animal