STREAM SAMPLING: Where’s the contamination coming from?


Is E. coli found in local waterways coming from humans or from animals?

Short answer: We don’t know yet.

Last spring, Brown County Health Department staff and volunteers visited creeks, ponds and lakes in select locations in Brown County to collect water samples. Those were sent to an Indiana State Department of Health laboratory in Indianapolis to analyze in an attempt to determine what’s in the water and where it’s coming from.

Most of those samples came from the Bean Blossom area, in areas the Brown County Regional Sewer District is planning to serve with public sewers to replace individual septic systems. Some samples also were collected from streams in Nashville in areas not currently served by public sewers. Sample collection took place in March and May, 2019.

The sewer board had hoped to see results and share them with the public early last summer. However, the process took longer than expected, and when results did start to appear, they were not as clear as board members had hoped.

“What we don’t have yet is an opportunity to sit down with the state lab director overseeing this and have them explain carefully and thoroughly to us what that tells us,” member Clint Studabaker told the sewer board meeting audience in July. “I don’t want to make assumptions at all.”

Sewer board members believed that this testing — checking for chemicals such as caffeine and nicotine, which would logically only come from humans — would help answer the big question: Whether the E. coli bacteria that other tests had found in lakes and streams was coming from animals, or from humans whose septic systems were leeching into waterways.

“I think they’re still trying to get to that point,” Studabaker said earlier this month about the state lab’s work. But that lab has been beset with technology and staffing challenges, he said, so they’re not there yet.

What the reports said

In November, the Brown County Democrat asked for and received copies of 14 chemical analyses of water samples from the Brown County Health Department. Some were pharmaceutical analyses, looking at medicines and other substances found in the water. Others were reports just on levels of E. coli.

You can read more details about those reports, and clarification questions we asked experts from the state health department and the Indiana Department of Environmental Management, in the box included with this story. (Scroll to the bottom to see it.)

Based on the explanation we were given from the state about how to read the reports, we ignored any results with a “less than” sign (<) next to them and focused only on the chemicals or compounds that were detected above a given reporting limit. However, since IDEM said that there were no standards or benchmarks for pharmaceuticals in surface water, it’s difficult to say whether the levels of any of them were “good,” “bad” or otherwise.

We also ignored the readings for the pesticide DEET, which the experts said was a very commonly found chemical in water samples for unknown reasons.

The pharmaceuticals found in these water samples were:


  • Behind Abbey Inn (now called The Yellowwood) at bridge: sulfadimethoxine (veterinary antibiotic) at 39 ng/L
  • Pond behind Abbey Inn: albuterol (asthma medication) at 53 ng/L; sulfadimethoxine (veterinary antibiotic) at 36 ng/L
  • Bridge by Brown County Highway Department garage (downstream from Greasy Creek Road): albuterol (asthma medication) at 140 ng/L; sulfadimethoxine (veterinary antibiotic) at 36 ng/L
  • Behind Brown County Inn (downstream of the Town Hill neighborhood, upstream of the town’s wastewater treatment plant): caffeine at 66 ng/L; naproxen (Aleve, pain reliever) at 5.6 ng/L


  • Ditch near Wagler cattle field on Gatesville Road: sucralose (an artificial sweetener) at 400 ng/L; sulfadimethoxine (veterinary antibiotic) at 32 ng/L
  • 100 yards west of 1943 Gatesville Road: caffeine at 75 ng/L; gemfibrozil (cholesterol medication) at 1.6 ng/L
  • North end of Woodland Lake: acetaminophen (Tylenol) at 35 ng/L; albuterol (asthma medication) at 10,000 ng/L; caffeine at 91 ng/L; cotinine (a metabolite of nicotine) at 17 ng/L; codeine (a narcotic pain reliever) at 220 ng/L; sulfadimethoxine (veterinary antibiotic) at 43 ng/L
  • South end of Woodland Lake: trimethoprim (antibiotic) at 6.6 ng/L; sucralose (artificial sweetener) at 390 ng/L; sulfadimethoxine (veterinary antibiotic) at 45 ng/L
  • Woodland Lake east of Bittersweet at pasture: albuterol (asthma medication) at 88 ng/L; codeine (narcotic pain reliever) at 62 ng/L; sulfadimethoxine (veterinary antibiotic) at 37 ng/L
  • North end of Little Fox Lake (sample taken March 27, 2019): norfluoxetine (psychiatric medication) at 34 ng/L; albuterol (asthma medication) at 14 ng/L; caffeine at 100 ng/L; codeine (narcotic pain reliever) at 5.6 ng/L; fluoxetine (psychiatric medication) at 20 ng/L
  • North end of Little Fox Lake (sample taken May 2, 2019): albuterol (asthma medication) at 36 ng/L; caffeine at 230 ng/L; diphenhydramine (Benadryl) at 190 ng/L; gemfibrozil (cholesterol medication) at 56 ng/L; codeine (narcotic pain reliever) at 180 ng/L; sulfadimethoxine (veterinary antibiotic) at 14,000 ng/L; warfarin (blood thinner medication) at 800 ng/L
  • South end of Little Fox Lake at culvert: albuterol (asthma medication) at 36 ng/L; caffeine at 170 ng/L; cotinine (metabolite of nicotine) at 9.5 ng/L; gemfibrozil (cholesterol medication) at 1.1 ng/L; sulfadimethoxine (veterinary antibiotic) at 23 ng/L
  • Old Settlers Road at State Road 135 North bridge: caffeine at 73 ng/L
  • State Road 45 at first bridge: caffeine at 89 ng/L
  • Railroad Road at 6472 by culvert: caffeine at 73 ng/L
  • North side of bridge at 135 North/Railroad Road: albuterol (asthma medication) at 32 ng/L; caffeine at 85 ng/L; naproxen (Aleve, pain reliever) at 5.2 ng/L; ibuprofen (Advil) at 45 ng/L

What about E. coli?

Last March and May, the health department also had water samples analyzed for E. coli bacteria at many of the same sites where samples were collected for the chemical analysis tests.

Drinking water must contain less than 1 CFU/100 mL of E. coli.

“Most surface water in Indiana would not meet this standard,” says an undated publication on water quality by Purdue University, “but compliance with the drinking water standard is not required because water is treated before it is used for drinking.”

Instead, all Indiana lakes and streams are to meet the standards for “full body contact recreation,” or swimming, Purdue says.

That standard is less than 125 CFU/100 mL, with no sample testing higher than 235 CFU/100 mL. “CFU” stands for colony-forming units, or bacterial cells. However, this is to be measured in five samples over a 30-day period, and the average of those samples is the number that counts.

The Brown County Health Department did not take five samples in 30 days at all of these sites. Seven reports from only one day of samples, in early May, were sent to The Democrat in November.

Those samples came from the stream behind the Abbey Inn bridge, the pond behind Abbey Inn, the bridge near the highway garage on Old State Road 46, the north end of Woodland Lake, the south end of Woodland Lake, Gatesville Road near the Wagler cattle field, and Woodland Lake east of Bittersweet at the pasture.

When asked last week, health department Environmental Health Supervisor John Kennard said he didn’t have the E. coli reports from the other seven locations which the lab had also analyzed for pharmaceuticals. The ISDH also was unable to locate them by press time for this story.

Of the seven reports the health department provided, four exceeded the swimming standard for the presence of E. coli. Those samples were from:

  • the bridge by the Brown County Highway Department garage on Old State Road 46 (365.4 CFU/100 mL),
  • the north end of Woodland Lake (686.7 CFU/100 mL),
  • Woodland Lake at culvert east of Bittersweet at pasture (2,419.6 CFU/100 mL), and
  • the ditch near Waglers’ livestock field on Gatesville Road (greater than 241,960 CFU/100 mL).

Other water samples have been collected and analyzed for E. coli from various locations around Brown County over the past 15 years or more, said Studabaker, board member of the Brown County Regional Sewer District.

As of last week, Studabaker said he had not had a chance to sit down and go through all of that historical E. coli sample data to see if there were any fluctuations or patterns, or if it matched up in some way with the pharmaceutical analysis work done by the state lab.

The Environmental Protection Agency and IDEM create a list of “impaired” waterways every two years. The most recent report, for 2018, included Bean Blossom Creek, Lick Creek, Bear Creek, Bell Creek, Hopper’s Branch and Dunnaway Creek, all in the Bean Blossom/Jackson Township area. In all listings, the reason for impairment was E. coli.

The 2018 impaired waters list included 9,431 waterways throughout the state. “High E. coli values are clearly not unusual in Indiana streams,” the Purdue publication said.

The danger of E. coli-contaminated water is the exposure to disease-causing organisms, Purdue says. Mostly, they enter the body through drinking or swallowing the water while swimming or playing in it.

So, what’s next?

Since the Brown County Regional Sewer District won a $118,000 Ready Communities Implementation Grant from the Regional Opportunity Initiatives in late December, it will be able to undertake a more comprehensive study with a different lab in hopes of answering the question of whether E. coli and other substances found in stream water are coming from humans or animals.

That study will take the testing a step further, checking water samples that contain E. coli for genetic markers that indicate what type of animal the bacteria came from — a human or some other species. It’ll also look at other data, like housing density, where livestock live, and the location of wastewater treatment plants, to try to correlate where it might be coming from, but not necessarily a point on a map, Studabaker said.

The sewer district is going to be hiring a multidisciplinary firm that will “have the wherewithal and technical expertise to begin to do some of those analytics. It’s just never been done before because it takes quite a lot of effort,” he said.

The grant also will fund a study of “options for improving wastewater management for residential homes and commercial operations throughout Brown County.” Currently, only Nashville, Gnaw Bone and Helmsburg have sewers.

The timeline for this new testing and analysis project isn’t firm yet. Read more about it in a related story in today’s paper.

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In November, the Brown County Democrat received pharmaceutical reports on water samples collected from 14 locations in the Bean Blossom and Nashville areas last spring.

To get help reading and understanding the reports, we sent them back to the Indiana State Department of Health lab which did the analysis, as well as to the Indiana Department of Environmental Management’s Watershed Planning and Restoration Division. We also sent questions to each agency to try to understand what this data said in plain English and to learn the circumstances under which these compounds could enter a water source.

Media relations staff from both agencies answered back in December. (Though emails were sent to specific staff members, the practice for both agencies is for media relations staff to act as the go-betweens for questions from the media.)

We’ll be posting all the water sample reports we received on, along with this information from experts on how to read them, in case you want to see them for yourself.

Clarification questions sent to Mary Hagerman in the ISDH lab:

Would you be able to explain any of the drug names listed on these reports in more commonly used language — for example, is “sucralose” an artificial sweetener?

Answer (from Jennifer O’Malley in IDSH’s media relations office): “Here’s a general description of some of the most common compounds we look for:

  • “Acetaminophen — Tylenol
  • Albuterol — asthma medication
  • Carbmazephine — anti-convulsant medication
  • Metformin — anti-diabetes medication
  • Cotinine — metabolite of nicotine
  • Naproxen — Aleve
  • DEET — insect repellant
  • Diphenhydramine — Benadryl
  • Sulfamethoxazole, Sulfadimethoxine*, Sulfamethazine — sulfa drugs, antibiotics
  • Gemfribrozil — cholesterol medication
  • Ibuprofen — Advil
  • Trimethoprim — antibiotic
  • Lincomycin — antibiotic (human and veterinary)
  • Paraxanthine — metabolite of caffeine
  • Fluoxetine — psychiatric medication
  • Sucralose — Splenda, artificial sweetener
  • Tylosin — antibiotic (mostly veterinary)
  • Warfarin — blood thinner medication”

*NOTE: Newspaper staff searched for more information online about sulfadimethoxine, as it turned up in multiple local water samples. All searches told us that this antibiotic is used in veterinary medicine.

What does the caret/less than sign (<) in the “Q” column mean?

Answer (from O’Malley): “Units are ng/L. This is nanograms per liter, which is parts per trillion.


analyte // result // Q

Acetaminophen // 7.5 // &lt;

DEET // 1200 //

“To read the reports, if there is a “<” less than sign in the Q (qualifier) column, then that compound/analyte was not detected at the reporting limit in the result column. In the above example, that means we did not find acetaminophen at all, or did not find it above 7.5 ng/L. We did find DEET at 1200 ng/L. The results are corrected for the initial volume used, which is why the detection limits vary from sample to sample.

“Note that in our experience, DEET seems to be ubiquitous, either because the samplers are using it out in the field, or it is present in many surface waters.”

Are there “safe” levels of any of these substances for water that people are either playing in or drinking?

Answer (from O’Malley): “ISDH is unaware of any EPA limits for any of these compounds under the Safe Drinking Water Act or the Clean Water Act. You may need to verify with the EPA.”

Questions sent to IDEM’s Angela Brown:

As a watershed expert, could you explain how common it is to find substances such as these in the amounts shown, and what the reasons could be for them showing up?

Answer (from IDEM’s Director of External Relations Sarah Bonick): “There have been studies in other parts of the country regarding the prevalence of pharmaceuticals and personal care products in streams, but IDEM is not familiar with any such studies in Indiana. Typically, pharmaceutical care products are related to wastewater discharges into surface water, as many of these chemicals are metabolized in the body and excreted in urine (for example, cotinine is a breakdown product of nicotine found in urine), or are flushed down the drain. The highest detections in surface waters are often associated with municipal wastewater treatment plant outfalls.”

Is there a “safe” level at which any of these substances are allowed to show up in stream water and not be considered a public health concern or danger?

Answer (from Bonick): “There are currently no surface water quality standards or drinking water maximum contaminant levels (MCL), or other ‘benchmarks’ for pharmaceuticals in surface water or drinking water.”


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