Penitentiary Glen

Diatom slides from core sample

This week in lab we extruded sediment from the core we collected at Beach City on October 10^th and learned how to prepare and examine diatom slides. The process of preparing diatom slides from a sediment core occurs in five steps: extruding the core from the collection tube, cleaning the sediment in nitric acid, rinsing and centrifuging the sediment, drying cleaned sample onto coverslips, and fixing dried material onto microscope slides.

Step 1: Extruding the core

extruding a core

For the sake of time, we only collected 4 samples from our core, but ordinarily an entire sediment core would be extruded and collected in 1-cm or 1/2-cm intervals. To extrude the core, the sediment must be pushed from the bottom up through the top of the collection tube. To prevent loss of sample, a plastic “stage” is attached to the top of the collection tube (fig. 1). This stage also allows for the sediment to be scraped into Whirl-Paks at desired intervals. Each Whirl-Pak was labeled with its corresponding measurement in the core (i.e. the top centimeter of sediment is labeled “0-1”).

Step 2: ‘Cleaning’ the sediment

plastic "stage" attached to collection tube

Approximately 0.5 grams of each sediment sample was mixed with 20 mL of DI water and 10 mL of nitric acid in a beaker and boiled on a hot plate until the solution reached a volume of ~10 mL. This step is necessary to remove organic matter from the sample. Because the inorganic sediment and diatoms cannot be separated in core samples, removing organic material makes diatom identification and count data easier to collect. At the end of this step, we are left with inorganic material (sediment + diatoms) and nitric acid.

samples boiled in nitric acid

Step 3: Rinsing the sediment

We did not complete this step in class, but the samples must be rinsed with DI water in order to remove the nitric acid. To do this, the sample is transferred to a Falcon tube and centrifuged at 2000 rpm for 10 minutes. The supernatant nitric acid is decanted and the tube is refilled with DI water. Next, the tube must be shaken to resuspend the sediment pellet. This process is repeated 6 times to completely remove the acid.

Steps 4 and 5: Preparing permanent mount slides

A solution of ~0.1 mL of cleaned sample + ~0.9 mL DI water was pipetted onto a coverslip and allowed to dry for 24 hours. Three dilutions of each sample were made and prepared slides were examined for concentration of diatoms. Dry coverslips were fixed to microscope slides using Naphrax permanent mountant. For the remainder of class, we looked at previously made permanent mount diatom slides and identified different genera based on morphology. 

Some of the common genera found were also known indicator organisms. Acidophiles: Eunotia, Frustulia, and Actinella, as well as Pleurosira, an indicator of elevated conductivity. Below are examples of each morphological group of diatoms: Centrics, with radial symmetry (most often seen in valve view); Araphids, without a raphe and often found in chains or star-shaped colonies; Eunotioids, acidophiles with an abbreviated (short) raphe along the valve mantle; Monoraphids (not pictured), with a raphe on only one of the two valve faces, Biraphids, with raphes on both valve faces (pictured: a "Naviculoid" biraphid with symmetry to both the apical and transapical axes); Gomphocymbelloids, with asymmetry to one or both axes (or symmetrical to both!), Epithemioid, with its raphe in a canal, Nitzschoid, with its raphe in a keel along one side of the valve, and Surirelloid, with a single raphe in a keel around the entire periphery of the valve face.

Asterionella, an araphid, and two (separated) centric
Stephanodiscus valves
 

Pinnularia, a biraphid

Eunotia, with an abbreviated raphe along the valve mantle

Cymbella, a biraphid with asymmetry to
both apical and transapical axes 

Epithemia, with a raphe in a canal

Surirella, with a single raphe around the periphery

Nitzschia, with its raphe in a keel 

Diatoms from Soil Cores

 

Diatoms on a Slide from a Soil Core

Background:

                  Diatoms are single celled organisms that have shells composed of silica. These incredibly diverse tiny organisms can be found in both freshwater and marine ecosystems. They are often studied as important indicators of past ecological conditions, because when the diatom dies, the silica shells remain in the sediment.  Because they remain in the sediment for long periods of time, the composition of diatoms found at a specific location in a soil core can tell researchers what conditions were like when that layer was deposited. Diatoms can indicate water salinity, temperature, and pH, as well as show nutrient and pollution levels in a system. For example, imagine a system that experienced no pollution until humans developed the surrounding area. The diatom composition before and after human development would differ. You would expect to see more pollution tolerant diatoms in the soil after development, and fewer pollution sensitive diatom species.

 

Taking a Soil Core at Beach City

Extracting a Core

                   Sediment core extraction required several pieces of equipment. Two boats were used to navigate the body of water (lake) in order to find a desired location for an extraction. Once a desired location was found, the boat remained stationary for a minute to ensure no sediment agitation occurred before taking a sample. Then, the core tube was slowly lowered into the water and forcefully pushed into the sediment, reaching a desired depth. The extraction of the core was performed by careful lifting of the sediment core tube, followed by a quick placement of a rubber stopper at the end of the tube. Finally, both ends of the core tube were plugged and taped to insure preservation of the sample. While, performing a sediment core extraction, it was observed that the bottom of the core was fairly dark due to anoxic conditions that are associated with deeper sediment layers. The top however, observed a brown-dark greenish color, indicating higher levels of oxygen. While extruding the sediment core, the pH of water was tested using a pH meter, which indicated a fairly neutral and acceptable value within the range of 6.5 to 8.  

  

Removing the Bottom Stopper and Inserting Plunger

 Creating Samples
                  Samples from a sediment core are important in studying the history of a desired body of water. They are great indicator of historical biodiversity which can be used to recreate past environments conditions. This information can be compared to the present biodiversity and further analyzed in order to grasp a better understanding of the changing environmental conditions such as the pH, as well as nutrient and pollution concentrations within the body of water. 

Extruding the Soil Core

                  In order to create samples from the extruded core, whirlpac bags were obtained and labeled to ensure accurate tracking of samples. Then, the water layer present inside the sediment core tube was expelled using a plunger- like tool that pushed the sediment from the bottom to the top. A specially designed tray was fitted on top of the sediment core which was used to concentrate the sediment samples into 1 cm intervals. The intervals were collected (by scraping the top) and stored inside designated whirlpac bags. All of the samples collected were then further analyzed by preparing microscopic slides in order to observe diatom biodiversity. 

This Plate Helps Measure Out 1cm of soil

Samples are Scraped into Whirlpak Bags for Sampling

 Preparing Slides

                  To analyze the diatoms in the sample, they must be spread out onto microscope slides. The first step in slide preparation is to remove as much of the organic matter and soil from your samples. This can be done by boiling a small about of sample in nitric acid, to break down organic compounds. After the samples are boiled, they are rinsed with water several times to remove traces of acid. The finished diatom mixture is cloudy and will need to be diluted farther before it can be made into slides. It is recommended that you make three separate dilutions of each sample while making slides. 

Boiling Samples to Purify Them

A Purified Sample: Ready for Dilution and Slides

                  The slides themselves are made by placing the sample onto a clean cover slip.  The sample should cover the entire cover slip and form a mound in the middle. These will need to sit for at least twenty-four hours, as they dry.  Once dry, the cover slips can be permanently mounted to a clean microscope slide.  Attaching the cover slips permanently allows researchers to preserve samples for later analysis.

Alison Demonstrates How to Correctly Make A Slide

Make sure to get a mound of water in the middle of your cover slip. It shouldn't spill onto other cover slips.

 Some of the diatoms found include:

Cymatopleura

Stauroneis

Stephanodiscus

Navicula

Pinnularia

Cocconeis

Cymbella

Aulacosara

Ctenophera

Puncticulata

Nitzschia

Diatoma

Rhopalodia

Eunotia

Frustulia

Asterionella

Platessa

Surirella

Gyrosigma

Enopnema

Pleurosira

Cymbella Seen Under the Microscope

A Visit to Triangle Lake Bog and Herrick Fen

Triangle Lake Bog - November 2013

Herrick Fen - November 2013

 
Located an hour south of Cleveland, there are two beautiful wetlands: Triangle Lake Bog and Herrick Fen. Both of these properties are examples of endangered habitats. Most of Ohio’s wetlands have been destroyed by development, while the wetlands that are left are continually threatened by eutrophication and suburbanization. Eutrophication threatens wetlands with large increases in nutrients from fertilizer run offs. The large inflow of nutrients disrupts the delicate balance of these systems.

Triangle Lake Bog - Early September 2013

Triangle Lake Bog was formed when glaciers moved across Northern Ohio approximately 25,000 years ago. As the glacier retreated it left behind heavy ice blocks that broke off and settled into the soil. After the ice melted, the depression filled in forming a kettle lake. The remnants of this kettle lake can be seen in the middle of Triangle Lake Bog. Bogs are a unique type of wetland because they lack water inflow and outflow; they are fed only through precipitation. The lack of water flow causes bogs to be acidic and low in oxygen. Nutrient levels are low in bogs because decomposition is slowed by the acidity and anoxic conditions. These traits that are characteristic of bogs make them the home of a number of unique species. 

Herrick Fen - Early September 2013

Herrick Fen was formed at approximately the same time as Triangle Lake Bog on top of a large gravel deposit left behind by the glacier. Fens are continuously fed by groundwater and water drainage from the surrounding areas. Because the fen has constant water influx, the soil is considered hydria (saturated and sometimes anoxic). Nutrient levels in fens are higher than in bogs because the mixing water has higher oxygen content and encourages more decomposition. Fens are also home to some of the special species found in bogs.

Tamarack is a common tree found in bogs and fens

Grass of Parnassus: a rare plant found at Herrick Fen

The field trip to visit these Ohio wetlands was filled with excitement and new discoveries, ranging from algae/diatom collections to discovering and learning about unique plant species and their preferred environments. Plants that were the center of attention during the trip were carnivorous plants. Due to their low nutrient environment, these plants have developed unique methods of acquiring nutrients. These plants compensate by consuming and breaking down animals such as ants and spiders as well as different types of protozoa. They each have a different method of trapping their prey. The types of carnivorous plants that were observed at Triangle Lake Bog and Herrick Fen were pitcher plants, sundews and bladderworts.

Close-up of the digestive juices in the pitcher plant

One of the most common carnivorous plants found in Ohio bogs are pitcher plants (Family: Sarraceniaceae). They are characterized by folding their leaves like a cup and forming a pitcher. The pitcher contains a sweet nectar juice that attracts many victims but at the same time has digestive properties. These victims crawl into the pitcher and once trapped, are unable to climb out. Over time, they are digested by the enzymes in the nectar juice and consumed by the plant. 
 

Pitcher plants at Triangle Lake Bog

Sundews (Family: Droseraceae) are carnivorous plants that are characterized by having specialized tentacles. The tentacles contain a gel like substance that will attract prey such as fruit flies, gnats and ants. After a prey insect gets stuck on the tentacles, the tentacles will wrap around the insect to prevent escape. The trapped animals will be dissolved by the enticing mucus and then absorbed as nutrients. 
 

Sundews grow in beds of Sphagnum moss

Close-up of Sundew: Red tentacles are visible

Another type of carnivorous plants that can be found in Ohio are bladderworts (Family: Utricularia). These plants are aquatic and are characterized by having a unique trap mechanism, which involves actively pumping water in order to initiate the trap. During contact with prey, a trap mechanism is initiated which will cause the plant to swell up due to active intake of water from the environment and at the same time suck in the prey. Once the prey has been engulfed by the bladderwort, they will be dissolved by the digestive secretions produced by the plant. 

 

Bladderworts (photo credit: plants.usda.gov)

These wetland habitats are home to so many unique plants that this blog is only able to highlight a few of them.  There are more beautiful things to be seen at these nature preserves. It is well worth the drive from Cleveland to visit them. It is important that people  visit these wetlands and appreciate their biodiversity. The general public needs to continue to educate themselves about the importance of these habitats and why they should be protected. 

For more information on these locations, visit:
Herrick Fen 
Triangle Lake Bog

For more information on the plants species above, visit:
http://plants.usda.gov/java/

Catch of the Day: The Fish Families of Ohio

Professors and students work together to identify a "mystery" fish.

This week’s lab involved taking a tour through the fish families of Ohio, guided by Dr. Sheil. We began by covering the basics of fish morphology. The first thing emphasized was the arrangement of fins and their proper titles, which included: the dorsal fin, caudal fin, anal fin, pelvic fin and pectoral fin. The bony supports within fins are fin rays, which are softer and more flexible, and spines, which are stiffer processes. The bony plate that covers the gills is called the operculum. The line running the length of the fish’s body is known as the lateral line, and it is a sensory organ used to detect pressure changes in water.

Basic external anatomy. (Photo credit)

Our lecture also covered the three basic scale types: ganoid, cycloid and ctenoid scales. Ganoid scales are the basal, or ancestral, scale type that has an upper surface covered in ganoine, which is a mineralized tissue comparable to enamel. Cycloid scales are a rounded type of scale, whereas ctenoid scales have more of an irregular shape.

Basal (heterocercal) and derived (homocercal) tail conditions. (Photo credit)

Another of the basic morphological characteristics discussed were the two types of tails. The ancestral condition is a heterocercal tail, which is asymmetrical, with the vertebral column extending into the larger lobe. Alternately, the symmetrical homocercal tail type is derived and seen much more commonly in the fish of Ohio.

Meaghanardo da Vinci puts the finishing touches on her scientific illustration.

After learning the basics we were ready to take a plunge through the different families of fish found in Ohio. We traveled to each of the different tables throughout the lab and sketched the preserved representatives of the families, taking note of particular morphological features that will aid in identification.

Petromyzontidae: Lampreys

Lamprey specimens. Note the buccal funnel (right), which these parasites use to attach to fish hosts, as shown in the center photo.  (Photo credit)

Lampreys are parasites of other fish. They use their buccal funnel (mouth), which is lined with cornified “teeth,” and rasping tongue to feed of host fish. Although lampreys have a mouth, they are jawless, so they lack an operculum. The lamprey’s overall morphology resembles an eel in the sense it lacks pelvic and pectoral fins. The dorsal fin is posteriorly displaced and is fused with the caudal and anal fin. The dorsal fin can be undivided, have a wide notch between the dorsal and caudal fins, or have a deep notch between the two fins. The condition of the dorsal fin and the anatomy of the mouth are often used to characterize species belonging to this family.

Acipenseridae: Sturgeons

Representatives from Acipenseridae. These fish have conspicuous bony plates on their dorsum, four conspicuous barbels on the ventral snout, and heterocercal tails. (Photo credit)

The sturgeon is a large, prehistoric-looking fish that has an ancestral, heterocercal tail. This fish has bony plates, or scutes, that are arranged in rows running the length of the body. Under the snout, one can see four distinct barbels and a tubular mouth used for sucking food items off the substrate. There are two species in Ohio, Acipenser fluvescens and Scaphirhynchus platorynchus. Scaphirhynchus platorynchus can be differentiated due to its long, slender tail and large, bony plates on the caudal peduncle.

Lepisosteidae: Gars

Members of Lepisosteidae. Note the slender snout and abbreviated heterocercal tail. (Photo credit)

Gars have a long snout that is filled with long, fang-like teeth, making them look fierce in appearance. Typically found in slow-moving and turbid tributaries, these fish turn right or left to grab prey as they swim through the water column. This fish has an elongated body that is covered with rhomboid, ganoid scales, which function as “armor-plated” protection. The dorsal fin is posteriorly displaced, which is often associated with “sprinter” fish or fish that swim near the surface. The operculum of members of this family has many bones on the “cheek” rather than just a singular opercular shield. It should also be noted the gars have an abbreviated heterocercal tail. The genus that we have in Ohio is Lepisosteus.

Clupeidae: Herrings and Herring allies

Alewife specimens. The vertical eyelid and the ventral row of keeled scales that help distinguish this fish. (Photo credit)

This “cute” family of fish has a few unique morphological characteristics that can help in identification. First, the dorsal fin is situated almost directly over the pelvic fin. Secondly, this family of fish has no lateral line system. Also, on the underside of the fish there is a row of keeled scales that resemble saw teeth. Furthermore, the anal fin is so broad and long it almost appears to interact with the caudal fin. A particularly interesting member of this group is the alewife (Alosa pseudoharengus), which has a “bizarre” eyelid that opens and closes vertically as opposed to horizontally, and is a dead giveaway for identification.

Salmonidae: Salmons, Trouts, and Whitefish

Examples of fish belonging to Salmonidae. In the photo on the left, the pelvic axillary process is highlighted. In the photo on the right, the adipose fin can be seen. (Photo credit)

These fish have small cycloid scales which give them a smooth appearance. Both male and female fish of this family possess a bilateral axillary process located above the pelvic fin. Cameron shared that he recently completed work for a research project that utilized this fleshy process for genetic testing. Researchers remove the process without significant negative impact on the fish by clipping it with a pet nail trimmer. Another feature of this group is presence of an adipose fin, which is a small, fleshy fin, composed of fat and located between the dorsal and caudal fin. The brook trout (Salvelinus frontinalis) and the lake trout (S. namaycush) are two examples of fish from this family that can be found in Ohio.

Esocidae: Pikes

Fish belonging to Esocidae are distinguished by their "duck bill" shaped mouths. (Photo credit)

These fish look distinct because their snouts resemble duck bills and are filled with large canine teeth. Similar to gars, the pike’s dorsal fin is displaced posteriorly because this fish swims near the surface and is a “sprinter.” These voracious predators rest in the grass, wait for prey, and then shoot out to grab them with their sharp teeth. Esox is the genus found in Ohio. Muskellunge and pike are typically found in large lakes, while pickerels live in small streams.

Catastomidae: Suckers, Redhorses and Buffalo fish 

The common carp (left), which is member of the Cyprinidae family, can be confused with Carpoides (right), which is a member of the Catastomidae family. The center photo shows the ventral view of both fishes. Note the striate, sucker-like lips of Carpoides and lack of barbels.

The most obvious feature of fish in this family is the subterminal mouth. The large and striated, sucker-like lips are used to adhere to substrate and suck up food, such as periphyton from rocks and logs. In Ohio, we can find the red horse (Moxostoma), the buffalo fish (Carpoides), and the common white sucker (Catastoma commersoni). We were cautioned not to confuse Carpoides with the common carp, which belongs to Cyprinidae. Carpoides superficially resembles a carp, but is distinguished by a nipple-like process on the bottom lip and a lack of barbels.

Ictaluridae: Catfishes, Madtoms

Madtom (left) and catfish (right) specimens. Note the small size of the madtom, as well as the unforked, rounded caudal fin. Alternately, the larger catfish displays the characteristic forked caudal fin. (Photo credit)

Members of this family usually have more than eight barbels around their mouth. An interesting feature lies within the fins, with both the dorsal and pectoral fins having a large spine. The spines are defensive weapons, which prevent fish from being swallowed by impaling predators. Dr. Sheil mentioned that people walking in lakes will sometimes accidentally step on skeletons of these fish and receive a painful puncture wound from the dorsal spine. These fish lack scales, but an adipose fin is present. Catfish (Ictalurus) and madtoms (Noturus) can be easily confused if one does not know what specific morphological features to compare. However, adult madtoms do not get much larger in length than the distance between your extended thumb and pinky finger, while catfish can grow to be quite large. Looking at the tail can also help one decipher between the two. Madtoms have an unforked, rounded caudal fin, whereas the caudal fin of the catfish is forked. 

Cyprinidae: Carp, Minnows, Daces, Shiners and Goldfish

A close-up of the tiny barbels of a "unknown" member of Cyprinidae, which we later determined to be a northern creek chub. The central stoneroller (Campostoma anomalum, left) and the fathead minnow (Pimephales) belong to Cyprinidae. (Photo credits: 1, 2)

Cyprinids have a more terminal mouth and in some species, the upper jaw has fleshy barbels that hang from it. Sometimes these barbels can be cryptic. One example of a familiar Ohio cyprinid is the common carp (Cyprinus carpio). The goldfish (Carassius auratus) is distinguished from the common carp due to a lack of barbels on its upper jaw. Another group within Cyprinidae is the minnows (Campostoma), which can be characterized by a large “C" shaped upper lip that causes a heavy overbite. Fathead minnows (Pimephales) have only a slight overbite, but very crowded scales on the dorsum.

Anguillidae: Freshwater Eels

American eel specimens. Note the continuous dorsal, caudal and anal fin. (Photo credit)

The freshwater eel has almost a serpent-like appearance, with pelvic fins absent and a fused dorsal, caudal and anal fin. This fish has scales so small it appears to be scaleless. On each side of the fish we find a single opercular opening. A freshwater eel species found in Ohio is the American eel (Anguilla rostrata).

Atherinidae: Sliversides

Silversides (top) can potentially be confused with a similarly colored Notropis (Cyprinidae, bottom). However, fish in Atherinidae are distinguished by their distinct "bird-beak" shaped mouth, large eyes, and serrated row of scales running the length of the body.

Sliversides resemble a miniature freshwater barracuda. These fish have a very flat head and back, along with a “bird–beak” shaped mouth. Sliversides also have very large eyes. Running down the length of the body is row of serrated scales that stand out against the conspicuous cycloid scales of the rest of the body. Sliversides also have two dorsal fins, the first of which is shorter and has six spines. In Ohio, we find the brook silverside (Labidesthes sicculus). When captured in a seine net, these fish are easily recognized by the silver flash on their side that reflects light.

Moronidae: White Basses

Specimens from Moronidae. Note the spine on the opercular flap. (Photo credit)

The fish in this group have two dorsal fins, the first of which has spines that are sharp and the second just has flexible rays. The key feature for this family is a spine that is located on the opercular flap. Members of this family commonly seen in Ohio are white bass (Morone chrysops), striped bass (M. saxatilis) and white perch (M. americana). Moronids can be distinguished from members of Centrarchidae by the condition of the dorsal fins, which as discussed below, are fused in centrarchids.

Centrarchidae: Blackbass, Crappies and Sunfishes

Fused dorsal fins are characteristic of many members of the Centrarchidae family. (Photo credits: 1, 2)

When it comes to members of this family, attention to detail is critical for identification. Counting the number of dorsal and anal fin spines as well as examining the size of the scales is very important. In Ohio, members of this family include blackbass (Micropterus), which have fused dorsal fins and small scales. The sunfishes also have fused dorsal fins, but larger scales than the blackbass. Finally, Ohio is home to crappies (Pomoxis), which have 5-8 dorsal spines and 5-7 anal spines.

Percidae: Walleyes, Perch and Darters

Morphological features such as obvious paired dorsal fins, a torpedo-like body, and spines associated with the anterior of the anal fin distinguish fish belonging to Percidae. (Photo credits: 1, 2)

Fish in this family generally have large, blocky heads and the body depth tends to be less than the head length. The overall shape of the body resembles a torpedo. These fish also have conspicuously paired dorsal fins and a spine associated with the anterior end of the anal fin. In Ohio, we find walleye (Stizostedion), which have a very flat belly and two dorsal fins. We also have darters (Etheostoma), which have very blunt faces and huge pectoral fins. Finally, we have the roughbelly darters and logperches (Percina) that have a more tapered face compared to darters in the genus Etheostoma, as well as significantly smaller pectoral fins.

Gobiidae: Gobies

The cup-shaped pelvic fin of the goby acts a suction cup and allows the fish to hold onto rocks.

This invasive group of fish is distinguished by its cup-shaped pelvic fin, which acts as a suction cup to allow them to withstand the current and sit in place on the benthos. As we learned in a previous lab, these voracious gobies will eat the eggs of other fish species in the time that it takes a fisherman to pose with the captured fish for a photo. There are strict restrictions on fishing activity due to the threat of goby predation on the eggs of native fish species.

Cameron labels new preservation jars to update the JCU fish collection as Alison enjoys the aroma of fish preservatives. Meanwhile, Chelsea digs deep for more unknown fishes and Kim supervises the search.

We concluded the lab by helping improve the JCU Biology Department’s fish collection by identifying and sorting "unknown" fish specimens. Using a dichotomous key specific to families of Ohio fishes, we were able to organize mixed collections of unknown fish into fresh and properly labeled preservation jars. 

A figure detailing different types of scale counts that can be performed to achieve more specific taxonomic classification. (Photo credit)

In addition to basic morphological features such as fin condition, presence or absence of barbels, tail type and snout shape, it was necessary to count scales to determine finer classifications of unknown specimens.


Fish Family List

Petromyzontidae: Lampreys

Acipenseridae: Sturgeons

Lepisosteidae: Gars

Clupeidae: Herrings and Herring allies

Salmonidae: Salmons, Trouts, and Whitefish

Esocidae: Pikes

Catastomidae: Suckers, Redhorses and Buffalo fish

Ictaluridae: Catfishes, Madtoms

Cyprinidae: Carp, Minnows, Daces, Shiners and Goldfish

Anguillidae: Freshwater Eels

Atherinidae: Sliversides

Moronidae: White Basses

Centrarchidae: Blackbass, Crappies and Sunfishes

Percidae: Walleyes, Perch and Darters

Gobiidae: Gobies