Dear Mrs Metcalfe,

please be so kind as to send me a bottle with some specimens in different size. I will try to pin and set them or find a way to prepare them so that they can be given to specialists in order to be identified. It is no good idea to throw away the material as it is a sample of biodiversity. I will do my best and write you my experiences. My address:

Dr. Thomas Witt, Tengstr.33, D-80796 Munich

With kind regards, Yours sincerely

Thomas Witt

If you have to go for biological extraction or isolation of most required biomolecules, in such a condition you have to remain careful and preserve the insects in cold by addition 1% formalin. You should try to store them in formalin at least for two days, you will get positive result. 

DNA-Analysis (see http://www.boldsystems.org/) is presently the most easiest way of identifying alkohol-preserved specimens and it may also become cheaper in the furure. For that reason, you should store the specimens in absolute ethanol as soon as possible after catching them and not do any other treatment. Do not add formalin or any other chemical and do not use methylated spirit. Such samples can normally be identified well, provided that they are not older than about 10 years.

Moths in alcohol can be stored for a very long time. Our Institute has a rich collection of immature stages of Lepidoptera is stored in alcohol. To determine the species on the genitals. Besides from the abdomen of females,stored in alcohol, you can remove the eggs and explore them. I believe that this material is of great scientific interest.

Of course, Lepidoptera can nearly always be determined when examining the genitalia. But this is really an exhaustive way with thousands of specimens. This is possible also after many decades or even centuries. In any case, the material is valuable and should be kept.

Certainly the material should be kept. As pointed out above, genitalia is useful for determining species, so one does not need specimens in perfect condition. What you have is base line data for what occurs in that area, it will certainly prove useful in the years to come! 

Yes, you can study this material with the genital slide, but it' s a big job.

Please, in alcohol only the inmature stages of Lepidoptera.

Please keep them all, meanwhile you can search for student or interested researchers to separate by morphos and pin a part of these. If you cannot process all the insects look for museums  or universities interested in having this material. Preserving a great part of it in absolute alcohol  will ensure future ADN  studies. Male genitalia is also very important to separate species. Although ADN is in vogue for this days, future generations should relate it with corresponding (must be) preserved insect morphology becuase ADN studies alone do not "tell all the story about animals".

 Thank you all for your input. The bottles that these specimens are currently in are not sufficient for long term storage - they are plastic and do not have gaskets. Ethanol is definitely evaporating from them and needs to be replenished every two years minimum.

What do you all think about storing the moths dry and flat in wax envelopes? I realize it ruins the possibility of DNA-analysis, but it would still allows identifications with genitalia and to get measurements for phenological studies. Has anyone ever seen leps stored this way?

A little more background: These moths are bycatch of a project that focuses on aquatic insects captured with light traps. Curating a high quality collection is unfortunately outside of the scope of my lab and my centers research goals.

Dried, papered insects can be kept for centuries in airtight boxes. If you take the moths out of alcohol, let them dry a bit and then, when they are still soft but not wet, place them in paper triangles and let them dry before sealing the packets, they will keep forever. All you need to do is add some napthalene in the box before closing it. Make certain the insects are perfectly dry before shutting the airtight box.

These can be used for genitalia studies and in many, there will be enough of the wings, etc left to place at least to genus level. Good luck!

Please transfer the sample to high quality air tight (rubber-ringed) storage jar with full data written on acid free paper in #2 graphite pencil.  Put the label inside he jar in he alcohol.  High quality jars obviates the need for regular replenishment of the alcohol, and preserves the samples for a very long time.  Proper labeling of each sample with the label, inside the jars, not on the outside, preserves that the data stays with the samples.  Find someone experienced with alcohol samples, such as Dr. Sandra Brantley at University of New Mexico in Albuquerque to determine the best long term storage containers, and the proper fluid.

Whenever possible, do not store lepidopterans in alcohol.  Finding someone to do all those dissections is nearly impossible.  Most of us (the number dwindles every year) who do dissections much prefer to work on specimens when we can see the wing patterns.  Genitalia are only part of a species identity. Whenever I'm offered alcohol stored lepidopterans, I reply "Maybe someday when I have nothing else to do."

And please, deposit the new and improved storage jars in a museum equipped to handle alcohol storage. jars.  Do not let the jars stay in a storage site where they will be overlooked by researchers.  OSHA, EPA, and other governmental agencies have strict requirements for storing alcohol samples. A large university or governmental museum is best. for keeping the samples. Storing the samples away in an obscure location thereby making them unavailable to researchers would be a tragedy.

Good luck.

Eric

My answer to your question is: Not much. My advice would be (based on experience with alcohol soups and also contact with lepidopterists) to dump the samples. Based on your description they are by now useless for either DNA extraction or for morphological study. I know of no lepidopterists who can use alcohol specimens that have been stewing in general samples (leps specially collected into 95% for DNA are another story).  Some of the suggestions from others in this forum may salvage some material if you have the time, space and money, but I suspect you don't have an abundance of those.

Another question is why are you mass-trapping at all? What do you hope to find in the aquatic insects that you can't find through less destructive collecting? In mayflies your sample will be largely subimagos which aren't much more use for identification than the moths in your moth soup. One method we use is put a small tray of alcohol on the ground with a coarse screen across it to keep out large moths and a UV fluorescent tube laid on top of that, connected to a battery with about two hours of life. This traps plenty of aquatics—you'll still pick up more caddisflies than you will ever classify in your lifetime—but also cuts down on the moth kill and spares all the large species.

My main suggestion would be to think carefully about the goals of this project and use more focused collecting methods to achieve them.

Dear Mrs Metcalfe,

please be so kind as to send me a bottle with some specimens in different size. I will try to pin and set them or find a way to prepare them so that they can be given to specialists in order to be identified. It is no good idea to throw away the material as it is a sample of biodiversity. I will do my best and write you my experiences. My address:

Dr. Thomas Witt, Tengstr.33, D-80796 Munich

With kind regards, Yours sincerely

Thomas Witt

Dear Dr. Witt,

I agree with you, that the material must not be thrown away. I therefore would like to offer my assistance with determining pinned specimens to family level and to make genitalia slides of non-ditrysian and tineid moths as far as possible. My address is:

Michael Kurz, Josef-Waach-Str. 13/1, A-5023 Salzburg

Kind regards

Michael Kurz

Dear Dimitri,

Please share the note with us when you are ready.

Thank you.

Eric

I agree with other kollegs that the material must not be thrown away. As  dr Witt says they are saples of biodiversity. Many of the species are in depression  for a long period of time and are difficult to collect. The meteorological conditions are different in every year as well. I hope this material will be very  use full for comparisons  the diversity of species.

Kind regards

N. Velcheva

It's possible to identify lepidoptera through a detailed analysis of its genitalia. The fauna of this region is possibly not rich in species and not studied and the genitalia is possibly represented in some paper. Thus the material collected can be valuable for biodiversity, phenological studies, population ecology, ethology and natural history. If possible, it's better to send to taxonomy specialists and try to publish the data. The data for Lepidoptera can be compared with other insect orders. First, the material can be separated in a number of morphospecies. It's desirable to set the identity to at least a family level. Than its necessary to prepare the genitalia or send a sample of the individuals of the morphospecies to a specialist or try to identify with literature. First. Even morphospecies level of classification is desirable as an estimated biodiversity index.

I answered you. I am a worker from Germany. You make a lot of blah blah internationally on research gate. I made a true offer to find a way to solve your problem. You do not hear this! Only international blah blah. Go on to so so, but without me!

best wishes Thomas

Dr. Witt,

I by no means ignored your offer. I have been away from the computer (holidays), and only read your response yesterday. I immediately began to discuss the potential of sending specimens to you and Dr. Kurz with my supervisors. I apologize for the delayed response. These specimens do not belong to me, I am helping with the management of them. I would definitely prefer to send specimens to interested researchers, especially to you and your collection which is a wonderful place for these moths, rather than to throw them away, but it will be several months before I could even get full permission to begin preparing these samples for shipment. This post was requesting advice, not offering specimens. Thank you for the idea, I am actively exploring it and will be in touch once we have a plan for these samples.

Happy new year,

Anya

Dear Mrs Metcalfe,

thank you for your answer. If you ned any help I will do my best. With my best wishes

Thomas Witt

To collect Lepidoptera for scientific purposes and store bulk collections in ethanol alcohol is plain stupid at the very least. You could have much better quality insect specimens if saved in 70% isopropyl alcohol. The whole assumption of using very expensive 95% - 100% ethanol was the wrong method from the beginning.  But still who has several lifetimes to look over almost worthless material. I am one of those taxonomic-minded persons that says never throw any specimens away, but in this case what you have done is wasted lots of time and energy, and alcohol and expensive plastic storage bottles and much needed storage space.   Either process Lepidoptera properly using established and proven methods, or not at all.  If they cannot be spread at the time of collecting, at lease a selected sample of hundreds should be pinned and bulk labeled with data, with the obvious duplicates discarded.  Otherwise this continued collecting and storage as you outline is worthless. The man-hours required to later sort through just a few jars of specimens with locked wing muscles, beat up and devoid of realistic color, and devoid of scales, is mind-boggling, not to mention investigating questionable specimens, as all of them will be questionable specimens.  How many hours will be required to identify 10,000 specimens of a single pest moth species ?  I would think it would take a staff of several hundred taxonomists. Using inexperienced students to sort this material would be an even more stupid exercise in worthless futility.  What would be the reliability of using students to identify species.

 I have collected insects 365-366 days/nights of every year for the past 46 years, in huge proportions (as many as 36,000.000 in a single day using hundreds of insect traps) and done so like this for the past 60 years, and sorted, processed and or discarded millions daily. But those I have culled from the masses of pest species and valuable rare and new species of Lepidoptera has amounted to more than a million nice spread, pinned, labeled, and mostly determined specimens.

I think the person who initiated this project was totally lacking in actual taxonomical understanding at your facility, and did not look at the bigger picture of what is obvious folly.  The ethanol you are using locks the wing and leg muscles of the specimens over time, so that they cannot be spread or pinned later.

There are many other insect orders which can be stored in 70% isopropyl alcohol, and I have done so, but there is some sorting on the front end, for example, here are some hemiptera, coleoptera, hymenoptera, etc., stored in 70% isopropyl alcohol.

If these moths are stored in 95% ethanol you may use them to various genetic analysis. My friends from the lab make such a diversity analysis. Please ask Piotr Gadawski or Michal Grabowski, here on RG, they will help you to use this wide collection for a good result and good publication.

 According to her initial posting, there would now be 6,000 light trap samples of lepidoptera, not coleoptera or other orders. This is very unclear as to what 'light trap' is being referenced , perhaps the old standard, a 15 watt fluorescent lamp trap, what if perhaps those light traps have 1500 watts of high intensity discharge lamps, along with (8) 15 watt fluorescent lamps on a single trap. The difference would be tens of millions of specimens for each trap sample.  No one with any common sense would consider spending the typical current cost of $10.00 US for DNA analysis on each specimen of alcohol stored lepidoptera.  DNA alone answers nothing, it is almost meaningless without accompanying undamaged quality adult specimens and availability for associated study of morphological, genetical, phenological, etc., etc., characteristics. To think otherwise is foolish, and knowledgeable taxonomist are well aware of this.  I previously stated having captured as many as 36 million insects in a single day using primarily light traps. Why would someone spend the huge amounts (hundreds of millions) of $$ to build and operate a facility, hire hundreds of taxonomist specialist, millions of $$ worth of laboratory equipment to sort through such worthless alcohol-stored material. Have you ever seen lepidoptera stored in OH for many years. The only things of any value whatsoever in an alcohol filled Nalgene bottle of lepidoptera, are the plastic bottle and the alcohol, both for use as fire fuel. Consider, one persons definition of what a 'light trap' is can be quite different from what someone else envisions when they see the words 'light trap'.   

https://www.academia.edu/9665262/Images_of_some_of_the_insect_traps_designed_fabricated_and_operated_by

If as indicated, these specimens came from the National Park, these discussions are interesting from an intellectual point of view, yet the fact remains the NPS owns them, and only the NPS can decide what to do with them.  Sounds like the NPS approved a not-too-well thought out experiment design, IOW what was the exit strategy? What will` happen to the specimens? What were they thinking about? I would love to have just the jar and alcohol budget.

I am doing a long term study of moths at White Sands National Monument. I put out 4 USDA-type bucket traps every month 12 months of the year This will seem paltry to Vernon who runs nightly traps. However I am required to sift through every single specimen that goes into the traps. I have to voucher every species of Lepidoptera, and, in the case of the microlepidoptera they are impossible to identify without making hundreds of dissections . In fact nobody can identify all the microlepidopterans without DNA bar odes or dissections. DNA bar coding is expensive particularly when because I am self-funded – that is why I spend many many hours every day dissecting microlepidoptera.. The upside is I discovered a great many new species in the National Monument. The downside is page charges are expensive. All of the duplicate specimens of moths go out onto my front yard where they become food for the lizards. This way of disposal of duplicates of common moths was all arranged with the National Park Service before I began the study.– I've been doing this for 10 years. The amount of material and processing` time is enormous.

Before I got started I laid out a clear plan of what would happen to every single specimen that ended up in the traps. I go through every sample and pull out every moth from all moth families. Everything else goes into 95% ethanol and is deposited at the Museum of Southwestern biology at the University of New Mexico. The moths are sorted to species, one of each species is vouchered, the rest become food for the lizards in my front yard.  in the case of microlepidoptera and they all look alike,  each specimen is vouchered.

Storing so many jars of alcohol specimens requires a lot of space. New Mexico State University, and the Museum of Southwestern Biology at the University of New Mexico agreed to take the alcohol samples. These two universities along with the AJ Cook Arthropod Research Collection at Michigan State University, where I have an adjunct appointment take spread and identified lepidopteria. The 2 universities in New Mexico utilize students to prepare the alcohol samples for future study. These two universities also agreed to take voucher lepidopterans. They both have storage issues to deal with. The National Park Service never thinks about how much it costs to store all this material that they claim to own. The National Park Service is very happy to let someone else pay those costs. That is why so many moths and up scattered on my front yard after I pick out the vouchers. There simply is not enough storage space to handle all material, and the lizards love it :-)

Because I do not recognize most microlepidopterans to species nearly all them must be dissected, A great amount of storage space is devoted to the specimens of microlepidoptera even though they tend to be quite small many drawers in my collections room are full of microlepidopterans from White Sands National Monument waiting to be dissected.

For me the bottom line of this give and take is t that he decision to collect all this material in the Grand Canyon was not well thought out without prior arrangements for storage and what to do with the samples. To be searching for the answer now has the cart well ahead of the horse. I hope the readers of this discussion are thinking about how they might approach a similar problem when collecting in a US National Park where everything collected belongs to the National Park Service.

Best wishes for the upcoming holiday season.

Eric H Metzler, winner of the 2016 Emil H Haury Award for outstanding scientific and scholarly contributions to Western National Parks and Monuments

Alamogordo New Mexico.