Living Material Replacement Questions

Living Materials Replacement Questions

Questions to Ask Before Processing a Replacement of Living Material

Check online Care Sheets for care information and FAQ’s.
General
• What indicates the material is not useable?
• What was the condition of the package when received?
• Always check tracking to ensure the material was delivered on time.

DOA & DAA
• When did you receive the order?
o Track the package and see if there was a delay
o If delayed, use CDL for replacement or credit
• Are all of the organisms affected, or only some?
o We send one or two extra with most of our organisms. So if customer ordered a pack of 3 and got 4 with one dead, no replacement is needed.
• Cold Weather
o Many of our organisms may look dead until they warm up – Let them come to room temperature first

• Dead After Arrival
o Were the organisms unhealthy when you got them, or were they fine and then died later?
o Have you reviewed our CareSheets and online videos? Would you be interested in speaking with tech support?
▪ To find a CareSheet or video, look up the item online, click on the Resources tab, and click on the link. Email CareSheet or link to customer directly if need be.
▪ For technical support: Sarah ext .4381, cell 263-2795 (sarah.bottorff@carolina.com); Adrian ext .6201  (adrian.yde@carolina.com)
o If the person calling is not the end user:
▪ Ask them to share the CareSheets and videos, and suggest that the end user can contact tech support (or tech support can contact them) if they have any questions

Omits & Empty Containers
• Omits
o Did customer receive other items with the same pick number?
▪ Check to see if the order shipped in multiple containers. Missing items may be in a container not opened or not yet received.
• Empty Containers
o Have you spread out and examined all the material in the container, paying special attention to any folds or crevices?
▪ Termites are often in the wood or in paper toweling
▪ Pillbugs are often hidden in folds of the paper toweling
Inside the container

Received too Early
• Determine if the CSR keyed incorrectly or if department shipped early
o If available compare PO to the Keyed Order
o If unavailable have phone call listened to
▪ Check if the dates keyed by CSR match the PO or customer’s requests
▪ If yes, then department shipped early (EAR)
▪ If no, then CSR keyed incorrectly (ISD)

Wrong Items
• What did you actually receive?
o Was the order keyed correctly? Compare our order to PO.
o Determine if they received the wrong item or if they received the correct item but it was mislabeled
▪ If they received the correct item, but it was mislabeled,
do not replace.
➢ If you are uncertain, call Sarah & Adrian.

Fish
• Did you have a tank set up before the fish arrived?
• What water source did you use?
o Tap water should be treated with a water conditioner to remove chlorine and chloramines – aging the water is not sufficient
o Bottled spring water is acceptable, but check the label to be sure it doesn’t have added vitamins
o Do not use distilled water
• See our CareSheets and videos on how to acclimate fish

Daphnia Magna
• How long after arrival, did you notice a problem?
o For most purposes, use Daphnia as soon after arrival as possible. We recommend within two days
• Did you add food pellets to the jar?
o This is unnecessary and often fouls the water, killing the Daphnia
• Are you trying to culture the Daphnia?
o Daphnia are difficult to culture. For most uses, it isn’t necessary to culture them. Use them for your lab or project as soon as possible

Termites
• Did you break open the pieces of wood to look for the termites?
• Because of their life cycle, termites are fragile in the late spring and early fall.
o We send extra termites during these times, but sometimes not enough survive shipment.

Drosophila
• For all cultures except Easy Fly
o Were the adult flies dead when you received the culture?
▪ Are there pupae on the sides of the vial and larvae in the food?
➢ If yes, check the date stamped on the label. New adults will begin emerging about two weeks from that date. If customer can wait, do not replace.
• For Easy Fly Cultures
o Were the adult flies dead when you received the culture?
▪ If yes, replace
➢ Easy Flies contain adult females only. They do not contain eggs, larvae, or pupae and must be used right away

Painted Lady Larvae
• Is the problem that you do not see the larva (caterpillars) moving?
o This does not mean the larvae are dead.
o Brush one gently with a soft paintbrush to see if it reacts
• The 30-35 larvae cup must be subdivided into smaller containers
o If not subdivided into smaller containers, they will die
▪ If they don’t have containers, refer them to 144068 (Culture Cup Set)
▪ If they don’t have food, refer them to 144040 (Culture Medium)
▪ If they don’t have instructions, refer them to CareSheet and videos

Elodea
• Is the problem that your Elodea is dark in color?
o Elodea is often very dark in winter. This does not indicate that it is bad
o If it is not falling apart or foul-smelling, it is good to use
• Did your Elodea die within a few days of receipt?
o Elodea will die or stop growing once it has exhausted the minerals in the water. Either do a partial water change every 3 to 5 days or use a water soluble fertilizer
• Why can’t you ship me Elodea densa (162101)?
o Some states ban shipments of Elodea densa. For those states we ship a substitute
o Refer to our sheet that identifies the substitutes (sent with each order)

Crayfish
• Are your crayfish dying or in poor condition?
o Crayfish molt in early fall and sometimes in early spring. They may not ship well during this time

Mantis Eggs
• Is the problem that your mantis egg case hasn’t hatched?
o Mantises may take 6-8 weeks to emerge from the egg case
o Recommend either 17291 (culture) or 172900 (kit) for food. The kit includes the culture

Tadpoles
• Are your tadpoles dying?
o Tadpoles have a high mortality rate even in nature. Out of 50, 2 to 5 might survive to become young adults
o Tap water should be treated with a water conditioner to remove chlorine and chloramines

Snails
• Are the snails not moving?
o This does not indicate that they are dead. Snails are often immobile for days.
• Are the snails floating?
o Snails often do this. It does not indicate they are dead. Dead snails seldom float.
• To determine if the snails are dead:
o Take a sniff. A dead snail has a really foul odor. There will be no doubt
o Pick up a snail; a dead snail will hang limply out of the shell, while a live snail will be tightly closed in the shell
• Snails are sensitive to heavy metals and other contaminants in the water.
o If your snails have died, try switching to bottled spring water. Make sure the water has no vitamins or other additives.

Amoeba Proteus
• Have you examined the jar under a dissecting scope (stereomicroscope) at 20 to 40X?
o Most amoeba will be on the bottom of the jar
o Amoeba move slowly. You may have to watch for up to 30 seconds to see any movement.
o Refer customer to our online video

Bacteria & Fungi
• What culture do you have?
• What is the lot number on the culture label?
• What indicates to you that there is a problem with the culture?
o If the culture is dead, what indicates that it is dead?
o If the culture is contaminated, what indicates that it is contaminated?
• How did you store the culture?
o Store at room temperature. Refrigeration can harm or kill most of these cultures.
• If they have tried to transfer the culture to fresh media (subculturing) and there is no growth on the new media:
o What media did you use?
▪ Recommended media are listed on the label, in our catalog, and in our online catalog
o How long have you incubated the new culture and at what temperature?
▪ Lower temperatures = longer incubation times. Recommended incubation temperatures are listed in our catalog and online. Most will grow at normal room temperatures but more slowly

Reactivating MicroKwik Cultures (both Bacterial & Fungal)

• Did you follow the instructions included with the culture?

• What media did you subculture on

•Recommended media are listed in a table on the instruction sheet.

 

NB 11/20/2023

Care guide

Caution
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This care sheet provides general information only for handling Carolina™ bacterial cultures. When you work with bacteria, it is imperative that you use sterile techniques at all times. Failing to use sterile techniques can contaminate cultures and work areas, and cause health and safety risks. See our Carolina™ Techniques for Studying Bacteria and Fungi Manual (item #154664) for descriptions of sterile techniques and standard practices for handling bacterial cultures.
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Immediate care and handling
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When your bacterial cultures arrive, immediately open the shipping container and remove the cultures. We ship cultures in tubes, plates, and as MicroKwik Culture® freeze-dried cells. Visually inspect each culture. Ensure that tubes are intact with caps securely in place, plates have no cracks and lids are secured by tape, and plastic bags containing MicroKwik Culture® cells are securely closed with the enclosed tube and vial intact.

You must seal any culture damaged during shipment—and anything it contaminated—in an autoclavable bag, and then sterilize it by autoclaving or soaking all contaminated materials in disinfectant. Note: Never discard a damaged, unsterilized culture in the trash. Know and follow your school or district’s guidelines for proper disposal. Contact our Customer Service at 800.334.5551 for a replacement of your damaged culture. It will help to have your order number available when you call.

Hold most cultures at room temperature, 20 to 22° C (68 to 73° F). For tube cultures, loosen the cap and keep the culture tube upright in a test tube rack or beaker. For best results, use cultures within 3 to 5 days of receipt; however, most bacterial cultures remain usable for up to 4 weeks when held at room temperature. Keep in mind that Carolina provides a variety of bacterial cultures and some have very specific care and handling needs that differ from these general guidelines. Vibrio fischeri, for example, must be kept in the dark and subcultured 2 to 3 times per week to maintain bioluminescence.

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Maintaining and culturing
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Eventually your bacterial culture will deplete the nutrients of the medium in or on which it is growing and will need subculturing. Before working with bacterial cultures, wash your hands with soap and water, ensure that the work area is draft free, and wipe the work surface with 70% alcohol or similar disinfectant. Note: Always check the culture for signs of contamination immediately prior to using. Never work in an area where food is prepared or consumed.

Transfer broth cultures to fresh broth using a sterile pipet or loop or streak onto agar using a sterile inoculating loop. For faster growth, you can incubate most cultures at 25 to 30° C (77 to 86° F). After making the transfers, clean the work area with disinfectant and wash your hands again. Either autoclave the old stock cultures and glassware, or cover them with disinfectant overnight. We recommend the use of Clavies® Autoclavable Bags (item #831642) for disposal.

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Biosafety
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In the US the CDC (Centers for Disease Control) sets standards for the safe handling of microorganisms according to their Biosafety Level (BSL). There are 4 Biosafety Levels with BSL-1 being the lowest risk microorganisms and BSL-4 being the highest risk. Bacterial cultures provided by Carolina Biological Supply Company fall into BSL-1 and BSL-2. Our catalog and online listings identify BSL-2 bacterial cultures as pathogens. They are only available as MicroKwik Culture® freeze-dried cells, and we can only ship them to colleges and universities. Cultures not identified in our listings as pathogens are BSL-1.

The CDC standards for working with BSL-1 microorganisms include the following:
[bulleted list]
Follow all standard microbiological practices.
Work can be performed on an open lab bench or table.
Wear personal protective equipment (PPE), i.e., lab coats, gloves, and eye protection, as needed.
A sink for hand washing.
Doors that separate the lab room from the rest of the facility.

For working with BLS-2 microorganisms the CDC recommends all of the above, plus the following:
[bulleted list]
Restricted access to the lab room while cultures are being used.
PPE to include face shields as needed.
All procedures that can produce a splash or an aerosol must be performed within a biological safety cabinet.
An autoclave or other method of decontamination for proper disposal.
Self-closing doors that separate the lab room from the rest of the facility.
An eyewash station.

For a fuller discussion of CDC criteria for handling microorganisms see the following link: http://www.cdc.gov/biosafety/publications/bmbl5/bmbl5_sect_iv.pdf [link].

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FAQs
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Which agar should I use for my bacterial cultures?
In Carolina’s print and online catalogs, the product descriptions for our bacterial cultures include the type of nutrient medium that we use for each culture. This information also appears on the label affixed to the culture. A wide range of bacteria will grow on Nutrient Agar (item #821862) and Tryptic Soy Agar (item #822022).

Can I pour my own plates if I don’t have an autoclave?
See our Nutrient Agar Bottle (item #776360) and other prepared media bottles. A bottle containing 125 mL of medium will pour 4 to 5 standard 100 × 15-mm plates. Watch our “How to Melt and Pour Agar Plates” video (at www.carolina.com) for a demonstration of the technique.
Should I order a tube, plate, or MicroKwik Culture®?
Tube cultures are best for stock. A tube culture is often streaked on a plate and incubated before lab use. Order a plate culture for convenience and immediate use. You can receive a plate culture and use it in a lab on the same day. A MicroKwik Culture® is best used for longer-term storage and to receive a culture of a pathogen. A MicroKwik Culture® can be held at room temperature for up to 2 months before being activated or for 6 to 8 months refrigerated.

Which cultures should I order for antibiotic testing?
Gram-positive and Gram-negative bacteria often give different results when tested against the same antibiotic. For that reason, we recommend testing against Bacillus cereus (item #154872), which is Gram-positive, and Escherichia coli (item #155068), which is Gram-negative. We recommend broth cultures because this makes it easier to spread the culture over the surface of an agar plate. See our Carolina BioKits®: Antibiotic Sensitivity kit (item #154740) for more information.
You recommend holding a culture at room temperature, but the recommended temperature for Escherichia coli (item #155065) is given in your catalog as 37° C. Why is that?
Thirty-seven degrees C is the incubation temperature, the temperature needed for maximum culture growth. The culture is mature when we ship it out and does not need further incubation. Maintaining the culture at room temperature allows you to hold it longer before use.

Care guide

Immediate care and handling
When your daphnia arrive, immediately:
Open the shipping container; remove and inspect the jar of daphnia.
Unscrew the lid and rest it on top of the jar to allow air exchange that is vital to daphnia survival. Note: Do NOT aerate the culture with a pipette. Do NOT screw the lid back on the jar.
Keep the culture jar in a cool area (21° C or 69° F) out of direct sunlight.

If you are planning on doing an experiment with the Daphnia, have them arrive as close to the procedure date as possible and have the experiment completed within two days. Establishing a long term culture is not necessary in most cases and we do not recommend it. If you are setting up a long term culture, see directions below in our “Culturing daphnia” section and allow at least one month to establish a culture.

Your shipment is a culture of at least 30 adult daphnia; juveniles may also be present. Do not be alarmed while inspecting the culture if you notice debris that looks like dead daphnia on the bottom of the jar. These are most likely discarded exoskeletons that daphnia shed as they grow and molt. Look for daphnia actively swimming slightly above the bottom of the jar, where they settle if they experience shipping shock. Give your daphnia 12 to 24 hours to recover and resume normal movement.

The culture should survive in the culture jar for 4 to 5 3 to 4 days as long as the jar's lid remains loose. For most uses, no further care is needed.

If you transfer daphnia to a different container, use only room temperature springwater. Slowly submerge the jar and pour its contents under the water into the new container. If you do not submerge the jar before pouring, air will be trapped under the daphnia's carapace. The organisms will then float, resulting in death. See the “Culturing daphnia” section below to maintain a culture of daphnia for more than a few days,

Culturing daphnia
We recommend culturing daphnia with 1 of our culture kits that conveniently includes materials and instructions you will need. However, if you want to maintain an existing culture of daphnia, there are several options for feeding it. We recommend our Daphnia Food (item #142316). Another good food source is a continuous supply of “green water” (algae-rich water), if you can provide it. Daphnia also feed on bacteria and yeast.

To prepare a yeast suspension, add springwater to a clean 2-liter plastic bottle and stir in enough baker’s or brewer’s yeast to make the water appear milky. Store the suspension in a refrigerator and always agitate before use to resuspend the yeast. Add a few drops of this food to your culture each day. An alternative is to crush 3 to 4 grains of dry baker’s yeast on clean paper and dust this on the culture water’s surface. Hard-boiled egg yolk or powdered egg yolk can be used in the same amounts as yeast to encourage bacterial growth. Note: Avoid overfeeding. If bacteria overgrow, they can kill the daphnia.

Do not allow the culture water to become cloudy. Each week draw off and discard about ¼ of the water; replace it with fresh springwater. Do not use city tap water because daphnia are extremely sensitive to the metal ions it contains. In fact, daphnia are so sensitive to metal contaminants that they are used to monitor the water quality of streams and lakes. Glass-distilled water or deionized water can be used if needed minerals are replaced. Brandweins and Chalkleys solutions are 2 widely used recipes for artificial liquid media.

FAQs
We just received our daphnia and they are all on the bottom. Are they dead?
Give them time to recover from shipping and begin swimming. Remember that daphnia grow by shedding their exoskeletons and these accumulate on the bottom of the culture jar.

I poured my daphnia into an aquarium and they all floated. What went wrong?
Daphnia have an expanded carapace that can retain air. When you poured the daphnia, air retained by their carapaces caused them to float and become trapped on the surface. Add daphnia to an aquarium by gently submerging the open jar and “pouring” them underwater.

We want to do a project on the heart rate of daphnia. Which daphnia should we use?
We recommend Daphnia magna (item #142330) for heart rate studies due to its larger size and slightly slower heart rate. For heart rate studies, we recommend that you use the daphnia as soon after you receive them as possible. See our Observing Daphnia Heart Rate video (www.carolina.com/video) for more information.

Which daphnia should we use to demonstrate feeding by hydra?
Use Daphnia pulex (item #142314) for feeding brown hydra. Daphnia magna is too large for the hydra to consume. You can also feed freshly hatched brine shrimp if you rinse them to remove the salt. See our Care and Handling of Hydra video (www.carolina.com/video) for more information.

How can I tell male from female daphnia?
During breeding season, this is easy. Female daphnia have a dorsal egg sac. Most daphnia populations contain females, with few if any males.

Can I keep daphnia in an aquarium with fish?
You can, but the fish will eat them. In fact, many aquarium hobbyists culture daphnia to feed their fish.

Can I refrigerate daphnia?
We do not recommend refrigerating daphnia. Keep them at normal room temperature and plan on using the culture as soon as possible after its arrival.

We tried to culture daphnia. At first everything went well but after several weeks they all died. What went wrong?
Daphnia populations sometimes die off. In most cases the females leave behind eggs that will hatch and restart the culture. It is best to maintain 3 or more separate cultures so you have a backup.

Care guide

The common fruit fly, Drosophila melanogaster, is the organism most widely used in genetic studies. It is also an excellent live food.

Immediate Care & Handling
When cultures arrive, remove the caps but leave the plugs in place. Keep cultures out of direct sunlight, and maintain at 20° to 25° C (68° to 77° F).

Culturing
It is easy to culture the fruit fly and its generation time is only 2 weeks at 21° C (70° F). The flies develop more slowly if you maintain the culture at a lower temperature, while keeping the culture at a higher temperature may promote male sterility, growth of bacteria and fungi, and mite infestation.

Use transparent vials (item #173120) or glass or plastic bottles (item #173135) as culture vessels for Drosophila. We recommend Carolina’s Formula 4-24® Instant Drosophila Medium (item #173200, #173202, #173204, #173218, #173210, #173212, #173214, or #173216). Follow the directions sent with the medium to set up vials for new cultures. Food medium should be moist at all times, as dry food will inhibit larval growth and result in few flies eclosing. Transfer flies to new culture vials every 10 to 14 days.

For more detailed information on setting up cultures and making crosses, see the Carolina™ Drosophila Manual (item #452620).

FAQs
Which type of Drosophila should I order to feed my mantis (or small frog or reptile)?
We recommend our flightless flies culture (item #144455). Because they cannot fly, they seldom escape from a terrarium or habitat with a fine mesh screen.

What materials do you recommend for culturing Drosophila?
Our Drosophila Culture Kit (item #173050) has everything you need except for the starter cultures. It also comes with the Carolina™ Drosophila Manual, which explains the procedures for culturing and doing Drosophila crosses, and gives information on crosses that are widely used in genetics labs.

Which cultures should I order for a beginning genetics lab?
In the Drosophila cultures section of our print catalog, the list of strains is color coded to indicate those with easy-to-recognize phenotypes that are good for beginning students. Among the most widely used are vestigial crossed with wild type to give a monohybrid cross; vestigial crossed with sepia for a dihybrid cross; and white crossed with wild type for a sex-linked cross. We list F1 cultures of these crosses for your convenience.

Care guide

Elodea densa is an aquatic plant widely used in biology for observation of plant cells and for studies of photosynthesis. This CareSheet covers care of Elodea for lab use. For use of Elodea as an aquarium plant, see our Aquatic Plants Carolina™ CareSheet.

Immediate care and handling

We ship Elodea (items #162101, #162102, and #162103) in a plastic bag without water. (The plant is also available as Elodea Tips [item #157340] in a jar filled with water.) Open the bag and examine the plants. Elodea can vary in color from dark brown to bright green. As long as the plants are not mushy and breaking down, they are healthy. Rinse the plants under running tap water, then shake them gently to remove excess water before use.

Care of the plants before your lab

Although you can use the plants immediately after rinsing them, it is usually best to transfer them to water and put them under lights for 2 days before using them in a lab. This allows the plants time to recover from any shipping shock. Shallow containers with a broad surface area are best. Use either tap water treated to remove chlorine and chloramines (see our “General Guidelines on Living Materials from Carolina Biological Supply Company” on www.carolina.com for details) or spring water. We do not recommend distilled or deionized water, as these lack minerals needed by the plants. Clip the rubber bands from the stems and spread the plants into a single layer in the water. A fluorescent light bank (such as our item #158999) or a 23- to 32-W compact fluorescent bulb in a desk lamp will provide the intense light needed by the plants.

If you must keep the plants for several days before lab use, replace about ¼ of the water every third day. This is to prevent mineral depletion.

Using Elodea to observe plant cells, chloroplasts, and cyclosis

Have students remove, with forceps, a leaf from near the growing tip of an Elodea plant. Place this leaf on a microscope slide in a couple drops of water and add a coverslip. Observe under a microscope at 40×, 100×, and 400×. Students will see cell walls and chloroplasts. They may notice that the chloroplasts of many cells are located along the inner cell wall. This is because a large vacuole occupies the central portion of the cell and tightly presses the cytoplasm against the cell wall.

Students will sometimes notice that the chloroplasts in a cell are moving. This results from movement of the cytoplasm, which is termed cyclosis. As the cytoplasm flows, the chloroplasts move along with it. Light and heat stimulate cyclosis. Tungsten or halogen substage lamps produce both heat and light, and after 2 to 3 minutes students will begin to observe cyclosis. If your microscopes have fluorescent or LED lamps, these produce very little heat and often will not stimulate cyclosis. To provide the needed heat, use a desk lamp equipped with a halogen bulb. Position the lamp so that it shines down on the lab bench. After a few minutes, the surface of the lab bench should become noticeably warm to the touch. Students can place their slides on this warm surface for 3 minutes and then observe for signs of cyclosis. A somewhat better arrangement is to position the lamp so that it shines directly onto the stage of a microscope, thus heating the slide as students view it. Although not all slides may show cyclosis, enough will so that students can share them and see the movement.

Substitutes for Elodea densa

Several states ban Elodea densa. If you live in one of those states, we must ship you a substitute. Here is some information on the plants we commonly send as substitutes.

Elodea canadensis (item #162111): This native plant is similar in appearance to E. densa but is a smaller plant and the leaves typically do not cluster as closely on the stem. Students can observe cells, cell walls, and cyclosis as with E. densa. E. canadensis is a good oxygen producer for studies of photosynthesis and can be shipped to most states. It is often unavailable in the winter months.

Elodea najas: Almost identical in appearance to Elodea densa and can substitute for all uses of that plant. Students can observe cells, cell walls, and cyclosis as with E. densa. E. najas is a good oxygen producer for studies of photosynthesis and can be shipped to all states. There are times when it is unavailable.

Chara (item #162120): The plants (Chara is a multicellular green algae) consist of stems with whorls of branchlets. It is a good oxygen producer for studies of photosynthesis. When viewed under magnification, the internodal cells (stems) show cyclosis; however, this is visualized as streaming of granules within the cytoplasm, not as movement of chloroplasts. The large chloroplast, which forms a background of green, does not move. Chara can be shipped to all states. Chara often has a strong odor, something like garlic. This does not indicate that the plants are dead or decaying.

Disposal

Elodea densa is a non-native plant that is invasive in many areas. After lab use, add leftover plants to an aquarium. Otherwise, seal the plants in a plastic bag, freeze, and discard the unopened bag in the trash.

FAQs

How can I use Elodea to study photosynthesis?
During photosynthesis, the plant absorbs carbon dioxide from the water and releases oxygen. Since oxygen is much less soluble in water than is carbon dioxide, the water quickly saturates with oxygen and bubbles form. The number of bubbles released per minute measures the rate of photosynthesis. Alternately, use a simple volumeter to measure the increase in volume of gas caused by the release of oxygen.

What is the difference between Elodea (item #162101) and Elodea Tips (item #157340)?
Elodea (item #162101) consists of Elodea sprigs that are about 11 cm (4¼”) in length. We recommend these for aquarium plants and for studies of photosynthesis, although they can be used also for cell studies. Elodea Tips (item #157340) are the terminal 3- to 4-cm ends cut from sprigs. We recommend these as best for cell studies.

Can I substitute other plants for Elodea densa?
For photosynthesis studies, almost any aquatic plant will work. These include Ludwigia (item #162141), Cabomba (item #162022), Myriophyllum (item #162161), Ceratophyllum (item #162041), and Sagittaria (item #162201). For easy viewing of plant cells with chloroplasts, consider Fern Prothallia (item #156877).

Problems?

We hope not, but if so, contact us. We want you to have a good experience.

Orders and replacements: 800.334.5551, then select Customer Service.
Technical support and questions: caresheets@carolina.com

Care guide

Salmonella precautions

Always wash your hands after touching an amphibian or any part of an amphibian’s habitat. For more information, see our “Amphibians, Reptiles, and Prevention of Salmonella Transmission” statement.

Immediate care and handling

Open the shipping container as soon as it arrives and acclimate the tadpoles to holding pails or habitats immediately.

Allow 50 to 60 minutes for acclimation. Have a habitat or holding pail made of glass, plastic, or stainless steel prepared with room-temperature water before proceeding. Use spring water, pond water, or dechlorinated tap water only. Spring water is available from Carolina (item #132458) or your local grocery store. If you buy spring water at the store, read the label carefully to be sure it contains no additives. If you use tap water, you must treat it first with a chemical water conditioner (such as item #971160, #672291, or #672292) to remove chlorine and chloramines.

To acclimatize your tadpoles:

1. Float the bag in the holding pail or set the bag next to the habitat.

1. After 20 to 30 minutes, remove about 1/4 of the water from the bag and replace it with water from the holding pail or habitat.

1. Wait 15 minutes and then repeat step 2.

1. After another 15 minutes, position a net over an empty cup or bucket (not the holding pail or habitat) and gently pour the tadpoles from the bag into the net. Transfer the netted tadpoles to the holding pail or habitat and discard the shipping water and shipping bag. Your tadpoles are now acclimated to their new environment.

Habitat setup and maintenance

Shallow trays, aquariums, and large culture dishes are all suitable tadpole habitats. Keep tadpoles in glass, plastic, or stainless steel only.

Clean the habitat with hot water before use, but do not use soap or detergent. Native tadpoles generally live in shallow water, so fill the habitat to a depth of 2 to 5 cm (3/4 to 2”). Bullfrog tadpoles can have a water depth of 8 to 13 cm (3 to 5”), and Xenopus tadpoles should have a depth of 10 to 20 cm (4 to 8”). Remember to always use spring water, pond water, or tap water that has been treated with a chemical water conditioner.

Change 1/4 to 1/3 of the water 2 to 3 times a week, or more often if it becomes cloudy. Be sure that any water you add matches the temperature of the current water. We do not recommend a filter, since it may suck up the tadpoles. A small aquarium pump with an air stone or other bubbler will help keep the water oxygenated. Keep the habitat at room temperature, away from heating or cooling vents, and out of direct sunlight.

Once a week, clean and rinse all habitats and utensils in hot water, without soap or detergent. Transfer the tadpoles to a holding pail while you clean the habitat.

Tadpoles can tolerate a wide range of temperatures but not a sudden temperature change. Never transfer tadpoles to water that differs in temperature by more than 1 to 1.5° C (2 to 3° F).

For maximum growth, put no more than 30 small tadpoles in every 4 L (1 gal) of water. As the tadpoles grow, decrease the population density by setting up more habitats and dividing the tadpoles among them.

Care and feeding

Most native tadpoles are vegetarian and live on a variety of plant materials. You can add aquatic plants such as Elodea (item #162101) to the habitat for decoration and as a food source. Provide enough light for the plants to carry out photosynthesis, but always avoid placing the habitat in direct sunlight.

Our Tadpole Food (item #146500) is an excellent food source for native tadpoles. Other options include pelleted rabbit food, fish food, and algae supplemented with finely powdered beef liver or powdered egg yolk. Parboiled lettuce and spinach are also suitable, but you should supplement them with other foods; tadpoles fed exclusively on lettuce or spinach may develop tumors. Xenopus tadpoles are filter feeders and will eat our Xenopus Tadpole Food (item #146501), nettle powder, or pea soup.

Do not feed more than the tadpoles can consume in a few hours. If your tadpoles are newly hatched from eggs, wait to begin feeding them until they are actively swimming. For 2 to 3 young tadpoles, a small pinch of food every other day is a good starting point. The amount you feed will depend on the size and number of your tadpoles, so it may require some trial and error. Increase the amount of food you provide as the tadpoles grow.

Remove any uneaten food from the habitat a few hours after every feeding. You may find that a pipette, turkey baster, or aquarium siphon makes it easier to remove the uneaten food, but be careful not to siphon up or injure the tadpoles.

After the front limbs appear, tadpoles may stop eating. This is because they are literally digesting their tails and need no additional food. Tadpoles also develop lungs at about the same time as their front limbs, and they will need a way to reach the air to breathe. Add a flat rock or other object to the habitat once the tadpole has hind limbs, so that it will be able to climb out as it matures. (Xenopus are completely aquatic, so they don’t need a way out of the water.) Frogs can climb on almost any surface, but toads need a surface that provides traction. Spring peepers are excellent climbers and will escape from any container that does not have a lid.

Frog habitat

Once frogs or toads are crawling out of the water with their tails mostly gone, move them to a terrarium. (Xenopus are completely aquatic; keep them in an aquarium as you would goldfish.) The terrarium should have a sand substrate and a source of water.

For frogs, put about 5 cm (2”) of clean sand in the bottom of the terrarium and bulldoze it to one end to create a land side and a water side. The depth of water depends on the size of the frogs, but it should be a few centimeters at most. Toads are terrestrial and can drown if they are trapped in water, so cover the bottom of their terrarium with clean sand and place a shallow dish of water on top. Keep the terrarium at room temperature and away from direct sunlight.

Native frogs and toads need live insects to eat. Start by offering them fruit flies, then add small cricket nymphs as the frogs grow. Larger frogs and toads can be fed redworms (small whole worms or chopped pieces), waxworms, mealworms, and crickets. Feed frogs and toads 2 to 3 times a week. Once or twice a week, dust the insects with a commercial vitamin/mineral supplement prior to feeding.

Xenopus frogs do not need live food. You can feed them pelleted food such as HBH Frog & Tadpole Bites (item #146503).

For additional information, see our Carolina™ Reptiles and Amphibians: Care and Culture manual as well as our individual Small Toad Carolina™ CareSheet, Tree Frogs Carolina™ CareSheet, and Xenopus Carolina™ CareSheet.

FAQs

How do I know if I have Xenopus tadpoles?

Unless you raised Xenopus tadpoles from the egg stage, you probably have tadpoles of a native frog or toad. Native tadpoles are heavily pigmented. Xenopus tadpoles, especially in early stages, are nearly transparent.

What species of tadpole does Carolina have?

We ship a variety of species depending on seasonal availability. In general, spring peeper tadpoles are available in the early spring, followed by toads and then grass frogs. In the winter we may send lab-reared grass frog tadpoles. Refer to any information that came with your order for the exact species you received.

Our tadpoles are now frogs. Can we release them into a pond?

No. A frog may be native to North America, but it may not be native to your area. Do not release Xenopus into the environment because it is not native and could damage native amphibian populations. Your state Department of Natural Resources or Department of Wildlife can advise you on relevant laws, guidelines, and regulations.

Our tadpoles are dying. What can we do?

Rushing the acclimation procedure can kill the tadpoles. Also, soap and detergent can leave a toxic residue.

You can try switching to a different water source. Tap water can have toxic metal ions, spring water from a grocery store might contain trace contaminants, and locally collected spring or pond water might contain a pollutant.

When performing water changes, be sure that the new water is the same temperature as the old water. It is also less stressful for tadpoles if you change small amounts of water more frequently, rather than changing a large amount of water all at once.

Unfortunately, the death rate of native frog tadpoles is often extremely high, even if you do everything right.

How long before the tadpoles become frogs?

Spring peeper and toad tadpoles transform into small adults in 6 to 8 weeks. Other species, including Xenopus, take from 10 to 14 weeks. Bullfrog tadpoles may take from 4 to 18 months to metamorphose.

Problems?

We hope not, but if so, contact us. We want you to have a good experience.

Orders and replacements: 800.334.5551, then select Customer Service.

Technical support and questions: caresheets@carolina.com

Care guide

Immediate care and handling

Open the shipment upon arrival, remove the container, and inspect your termites. Examine the moist paper towel and wood in the shipping cup. You will find the termites in the folds of the paper and in or on the wood. If you plan to use the termites within a few days, they'll need little or no further care.

Maintenance

We do not ship the reproductive castes of termites, so we do not recommend trying to establish a self-sustaining colony. Instead, your intent should be to maintain the shipped termites until you use them for either a lab or live food. You can maintain them in the plastic shipping cup for these purposes. We ship collected worker and soldier termites of various and unknown ages, so expect some natural, gradual die-off as individual termites live out their life expectancies.

Controlling moisture is critical. A dry culture soon dies, so check moisture at least daily. On the other hand, a water-saturated culture produces foul conditions unsuitable for termites. It is best to sprinkle a little water onto the top paper as needed to maintain a slightly moist condition. Note: Do not use tap water for this. Use water from an established aquarium or springwater (preferred).

Keep the lid tightly closed and keep the container in a dark, cool place. If mold appears, increase ventilation and cut back on adding water. Remove and replace any paper or wood that becomes covered with mold. Any well-rotted hardwood (oak, maple, elm, hickory, etc.) is suitable as replacement wood.

To dispose of termites that remain after your intended use, freeze them in their container for a week and then place the unopened container in the trash.

FAQs

If the termites escape, will they destroy the furniture in my classroom or damage the school?

No. They won't eat your classroom furniture if they escape. These termites belong to the genus Reticulitermes of subterranean termites that live in colonies in the soil. They infest materials that are in direct contact with some source of moisture, which isn't usually the case for most classroom furniture.

Do termites bite or sting?

Soldiers are capable of biting due to their large mouthparts. However, they won't hurt you since they don't have venom in their bite and can’t sting. At most, their bite produces some irritation.

I want my students to do the activity in which termites follow the mark of an ink pen. Do I order workers or soldiers?

We recommend workers for this activity. Order 100 worker termites for each class of up to 30 students to be certain that you have enough termites. You will find all materials for this activity, including worker termites, in our item #143722 Termites Catch the Scent! kit.

Why did my termites die?

The most common cause is too little moisture. Each day lightly sprinkle water on the top layer of paper. Keep the culture away from direct sunlight. Termites do best at temperatures below normal room temperature, with 10 to 15.5^° C (50 to 60^° F) optimal. In early fall, there may be large numbers of nymphs in termite colonies. Nymphs don't ship as successfully as adult workers and soldiers. We compensate by sending more termites in each shipment. During these times, you may see many dead termites, but there should be plenty of live ones for your use.

Problems?

We hope not, but if so, contact us. We want you to have a good experience.

Orders and replacements: Call 800.334.5551, then select Customer Service.

Technical support and questions: E-mail caresheets@carolina.com.