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INTRODUCTION

 

Welcome to the Proyecto Primates Protocol Site!  In the following pages you will find detailed information on the different protocols that are being used for data collection, as well as protocols that are necessary for the management of the project and for collecting samples both in Yasuni National Park, Ecuador and in Formosa, Argentina.  Anyone working in collaboration with Dr. Anthony Di Fiore and Dr. Eduardo Fernandez-Duque needs to read and follow the protocols.  It is the information  in the protocols that you are responsible for following, not the information passed to you by other colleagues, students, assistants or even Tony or Eduardo.  If you notice a discrepancy between what Tony or Eduardo tell you and the written word in the protocols, please bring that up to our attention so that we can udpate the protocols accordingly. 

ECOLOGY

BEHAVIOR

PROJECT MANAGEMENT

SAMPLES

Climatological Data

 Behavioral Data

Flagging, Trail Maintenance, and Mapping

Fecal Samples for Hormone Extraction

Fruit Morphology Data

 

Laboratory Maintenance and Equipment Care

Fecal Samples for DNA Extraction

Phenological Data

 

Safety in the Forest

Proyecto Mirikina's Information Package

Urine Samples for Hormone Extraction

 

 

Darting and Capture

Plant Voucher Specimens

 

 

Data Entry and Management

Plant Nutrient Samples

 

 

Position on Data Property and Signature Page

Salt Lick Camera and Video Camera Trap

 

 

Institutional Liability Release and Signature Page

 

WHAT IS A PROTOCOL?

CHARACTERISTICS OF OUR PROTOCOL

TYPES OF DATA TO BE COLLECTED

DESCRIPTIONS OF FORMS, SUBFORMS AND TABS IN DATABASE


WHAT IS A PROTOCOL?

            A protocol consists of a detailed description of the methods used to collect data. The reality is that, no matter what anyone might say, no one can read a published scientific paper and precisely replicate the described methods, because the methods published in articles are not sufficiently detailed to allow that. But, everyone should be able to replicate the methods of a study by following the “protocol” used for the study. The use of protocols is common in laboratory settings, where it is important to have explicit instructions. For example, a protocol would indicate if you should centrifuge a reaction for 10 or 12 seconds. Although protocols do not appear to be used that often by field biologists, we are convinced that they are extraordinarily useful.

            The secret to checking if a protocol is complete and ready to be followed is to “visualize” the entire data collection process: close your eyes, think about what you are going to do step by step, and make sure the protocol details that process precisely. For example, you are watching an animal and the alarm on your watch beeps: What do you see? What data you write down?  In formulating a protocol, you need to be very explicit and leave little in doubt or ambiguous about what data an observer should be recording. Never think, “It’s obvious”! Instead, try to leave a minimum of things open to the interpretation of the observer.

            In this project, you will be part of one of the first studies (we think THE first study!) where researchers are trying to collect truly comparable data on four different species of primates in two different countries with a set of unified protocols. The emphasis of our protocol is on the QUALITY of the data we aim to collect, not the quantity.


CHARACTERISTICS OF OUR PROTOCOL

            Our sampling procedure is designed so that all data can be collected by a single person. There is a lot of different kinds of data to collect, and we expect it to be collected, recorded, and entered into the computer in a very specific way, so you must reread the protocol pertinent to your work once a month during the first week of the month to make sure you remember everything you need to be doing.

            Our protocols have been designed to ensure that the collection of CERTAIN kinds of data is done in an exactly comparable manner for all primate species being studied both in Ecuador and Argentina.  There is another set of data that are being collected so that they are totally comparable for the four species under investigation. Clearly, it is impossible to collect ALL kinds of data using comparable methods, given for example the nocturnal habits of owl monkeys and the intrinsic habitat differences between the Amazonian rainforest and the gallery forest of the Argentine Gran Chaco. Nonetheless, our aim is that certain kinds of data, and the analyses to be performed on these data, will be COMPLETELY comparable

            To accomplish this goal, is of paramount importance that: 1- the protocols take precedence over what other students, assistants or even researchers may say.  You are responsible for reading the protocols and following them.  You must not assume that what you are being told is correct, 2- all observers need to practice inter-observer reliability or go through procedures together.  Once per month, at the beginning of the month, all observers should go to collect FOCAL SAMPLES together on the same animals, simultaneously. Data for practicing inter-reliability should be collected in data books or on the Palm, but you should note in the Comments section for that FOCAL SAMPLE that the sample was one where inter-observer reliability was being evaluated. When that data is entered or passed into the Proyecto Primates Master Database, mark the checkbox for Inter-Observer Reliability and note which observer collected the “real” focal data in the Comments section. That way the various data from the various observers collecting data simultaneously can be looked independently of other data being analyzed.


TYPES OF DATA TO BE COLLECTED

            There are a number of different types of data that you will be recording in the field and/or laboratory. The data we are collecting is organized in a hierarchical or nested fashion. All “inner level” data pertain to corresponding higher levels, but not all “higher level” data have associated “inner level” data.  Below you will find a table with all the different protocols that you may need to become familiar depending on the specifics of your work.  Regardless of the kind of data you will be collecting, there are three numbers that are extremely important and need to be entered consistently everywhere: OBSERVER SAMPLE NUMBER, AVISTAJE NUMBER AND FOCAL SAMPLE NUMBER.  WRITE THE OS, AV, AND FS NUMBERS IN ALL PERTINENT PLACES IN DATA BOOK (i.e., observer data should have OS number; avistaje data should have OS and AV number; focal samples should have OS, AV, and OS numbers). Detailed instructions for each of those follow. 

 

 

OBSERVER SAMPLES


            OBSERVER SAMPLES serve to organize almost all of the data being collected by each individual person on this project and also gives us an idea of how observers are spending their time and of what parts of our protocol are difficult and may need modification. This constitutes the highest level of organization of the database, and EVERY person affiliated with Proyecto Primates in Ecuador or Proyecto Mirikiná in Formsa must fill out an Observer Sample record for each day you are associated with the project.  Ideally, we would like this data for everyone who works on the project for more than a few days (e.g., volunteers, SIT students) BECAUSE this data can be used in grant applications (e.g., NSF) to QUANTIFY the Broader Impacts of our work in terms of training students, etc.

            This is simple and MANDATORY – these data MUST be collected each day, and there is no excuse for not doing it. The database has been reorganized so that this information must be added. Almost everything else in the database is pegged to a particular OBSERVER SAMPLE.

 

      In Ecuador:

In Argentina:

You MUST review your Observer Samples after they are entered (especially if you are uploading them from the Palm!).  Every Observer sample MUST have the following fields filled out:

Observer Sample Number

Observer

Date Dates in your book and in the database should all have the same format: DD-MMM-YY, e.g., "21-JUL-06".

Time Start AM

Time Finish AM

General Activity AM

 Time Start PM

Time Finish PM 

General Activity PM

      If you do the same activity all day, that should be entered TWICE, once for General Activity AM and once for General Activity PM.  The Time Finish AM should then be entered as noon and the Time Start PM should also be entered as noon.

      The fields "Avistaje Link", "Obs Activity Link", "Weather Link", and "Phenology Link" should ALL simply match the "Obs Sample ID" -- these fields are redundant in Access but are used by HanDBase to link tables.   These fields are not seen in the forms, only in the tables.

NOTE: you do not need to enter any more detail on your observer activities other than your general activity in the Morning and Afternoon!  This was an onerous thing that we were doing last year, but we have gotten rid of it.  However, the old data remains in the database in the Obs Activity Data table.

 

 

FOCAL SAMPLES

 

 

 

AVISTAJES

 

            Most of the data you will collect in the field are organized around AVISTAJES (“sightings”) or encounters, either with your focal group of animals or with other groups or species. These data are to be collected in the Avistaje section of your data book or the Palm data logger and entered in the Avistaje SubForm of the Database.

            You must collect AVISTAJE data EVERY TIME you encounter monkeys of any species, no matter whether you collect any other kind of data on them that day. This includes the animals you see while doing any field work activity. Every separate encounter you have within a given observer-day in the field constitutes a new AVISTAJE. For example, suppose you are with a group of sakis in the morning and you go back to camp for lunch (which should be rare!) or heavy rains force you to stop data collection. If you then return to the same group and do more sampling in the afternoon, the afternoon sampling session is considered a new AVISTAJE (since your original encounter “ended”) and the session should get a new number when the data are entered into the computer.

            How much data you will collect and the time spent in collecting AVISTAJE data will depend on the situation. For example, in Ecuador, if you are working in the Monogamous Primate Project AVISTAJE data should be collected as completely as possible for Pithecia, Aotus, and Callicebus.  For all other species you must get the time when you see them and an approximate location. All other information should be collected or not depending on how it will interfere with your current activity.

If data are not available for a given cell, enter "Not Available", but do NOT leave those cells blank.

            If you are entering data only on the Palm, be SURE you review your Avistaje data immediately after you have transferred it to make sure you have entered EVERYTHING that needs to be entered (i.e., that none of the required fields are blank or contain odd pieces of information.  Do not simply upload and forget about it!  Do this BEFORE you delete the data from your Palm.

            The fields "Range Data Link", "Focal Sample Link", "Biological Sample Link", "Feeding Bout Link", "Marked Tree Link", and "Ad Lib Data Link" should ALL have simply match the "Avistaje Sample ID" -- these fields are redundant in Access but are used by HanDBase to link tables.   They are not shown in the forms, only in the tables.

 

 

DESCRIPTION OF FORMS, SUBFORMS AND TABS IN PROYECTO PRIMATES DATABASE

            
    The Field Data Entry Form in the Database has the following five subforms in the following order from left to right: Avistajes, Phenology, Biological Samples, and Mapping Data.  In the following pages we provide detailed instructions on how the data you collect MUST be entered in the various subforms.

    There are other kinds of data that are not entered in the Proyecto Primates Database: FRUIT MORPHOLOGY (see the Fruit Morphology Data Collection protocol) and CLIMATOLOGICAL DATA for the site, including temperature, humidity, and rainfall (see the Climatological Data Collection protocol).

   

AVISTAJES SUBFORM:


    For each AVISTAJE associated with a follow of a focal group of animals, there will be a number of additional types of data you will be collecting, including Demography and Census Data, Ranging Data, Marked Trees Data , Feeding Bouts Data, Focal Samples Data, and Ad Libitum Data.

The Avistajes subform has six Tabs that address each of the different kinds of data listed below:

    Data on group size and composition and on presence/absence of known group members (Demography and Census Data).

    Data on ranging patterns, habitat use, general group activity, and weather conditions (Ranging Data).

    Data on the locations, identity, and collections made for feeding trees (Marked TreesData).

    Data on diet and feeding bout characteristics (Feeding Bout Data).

    Behavioral Data, including systematically collected data (Focal Samples)

    Opportunistic observations (Ad Libitum Data).

Demography and Census Tab:

    

    Data on group size and composition and on presence/absence of known group members.

Ranging Data Tab:


Throughout a follow, you will record the location of the group being observed every 20 minutes  on the hour and at 20 and 40 min past the hour  even if those points fall during a FOCAL SAMPLE. If you cannot take the position during the FOCAL SAMPLE, try to remember where you were and take the position after the end of the focal sample.

Some of the ranging data are recorded with the AVISTAJE for the group you are following since the AVISTAJE asks for the time and position where you first encounter monkeys and where you leave or lose them. The additional data on group locations recorded during a follow are recorded in the Ranging Data section of your data book or the Palm data logger.

Positions should be noted as distances and angles from reference points on the trails and transects or trees that have already been mapped. Whenever you can, it is better to take a position using a trail point rather than a tree as a reference point, but a nearby tree is better than a much further trail point.

All location data should be recorded in your data book in the form of “Distance, Compass Angle, Reference Point”. Do NOT record location data in any other way! Use the codes “m” for meters and “d” for degrees without inserting spaces after the commas. Example: “09:20 20m,250d,AV-05-021”. You should practice estimating distances as you walk on the trails, which are marked at regular intervals. Count your paces for 25 meters and then use that to guide your distance estimates. You will get pretty good at estimating distances of up to 40 or 50 meters using your paces! From time to time, bring out the laser range finder and check yourself!

IMPORTANT NOTE: When you note an angle, the angle should be AS IF YOU WERE STANDING AT THE MAPPED REFERENCE POINT. This means that if you take an angle from where the monkeys are back to a mapped point, you will have to invert the angle (e.g., an angle of 180 degrees read from where the monkeys are back to a reference point corresponds to an angle of 360 degrees from the reference point).

When you enter the data in the database you can enter ranging data comments. You should classify each ranging data point as one of the following:

We cannot stress enough how absolutely critical the accurate and systematic collection of the ranging data is for this project. You should not be missing more than a few ranging points in any given full day follow. Be especially conscientious of writing down the CORRECT tree number if you use a tree as a mapping point. It is very easy to accidentally invert two digits or to assume you know the tree number without looking at the metal tag, but this causes a lot of problems if you are wrong, because such errors are VERY HARD to correct post-hoc. If you are far away from a reference point or simply cannot stop to take the ranging data, and especially you are in a seldom used part of the group’s range or far away from trails, the ranging data are especially important. In this case, you may need to put up a temporary flag with the date and time and find your way back to that spot on a different day to map it. When you do this, be sure to go back and write in the data in the APPROPRIATE PLACE IN YOUR DATA BOOK or Palm. Please do this sparingly, however, and REMOVE the temporary flagging once you go back to map the point.

In addition to collecting ranging data every 20 minutes, you should also note the predominant activity of the group at that point as either “Resting”, “Foraging”, “Moving”, , “Socializing”, “Vocalizing”, “Other”, “Active Fuera de Vista”, “Inactive Fuera de Vista”, “Unknown” and “Not Recorded”. 

 

You will also use these points to keep an informal running record of weather conditions, which we can use to examine the relationship between weather variables and particular behaviors such as army ant foraging by saki monkeys. There are two different weather variables to record at the 20 minute point: Light/Rain conditions and Wind conditions. Light/Rain should be classified on a scale representing the degree of cloud cover and rain: “Rain”, “Drizzle”, “Cloudy”, “Partly Cloudy”, “Slightly Cloudy”, “Full Sun” and “Mostly Cloudy” for daytime observations, “Clear Night”, “Moon Visible”, “Moon Partially Visible”, “Moon Not Visible” and “Dark” for nighttime observations. Wind condition is classified on a 3-point scale: “No Wind”, “Slight Breeze”, and “Windy”.

Note that taking weather data this way during AVISTAJES supercedes our old way of collecting weather data all of the time during OBSERVER SAMPLES and should be easier to do consistently.

 

How to enter the data:

REMEMBER, the goal is to automate the assignment of actual X,Y coordinates to your ranging points... we cannot look up the X,Y value for a reference point that does not exist in the database (or that cannot be found), thus the above conventions are ESSENTIAL to successfully search the database for reference point location.

 

Marked Trees Tab:

 

            To continue gathering data on the diets and ecological strategies of the taxa we are studying, we mark and map the locations of feeding trees and we estimate the time spent in feeding patches of various species. Thus, each time you find your target taxon feeding in a food patch, you will mark the tree and record a number of pieces of data concerning the feeding source. You only need to mark only patches fed in by a group or individual monkey for more than five “group-feeding” minutes (i.e., “major” food sources: those fed in by a single animal for 5 minutes, or by several animals for a total of 5 “monkey minutes” over the course of the day). You should train your ears to listen for the sound of falling fruits and you should get in the habit of looking at your watch when you first hear fruits dropping in order to accurately assess whether or not to mark a patch.

            You will mark with plastic tape all those trees used for foraging as described above plus trees that you know the animals sleep in (sakis, titis, and owl monkeys all reuse at least several sleeping trees). Mark sleeping and feeding trees (or the trunk supporting the feeding source in the case of epiphytes, hemiepiphytes, lianas, vines, or bromeliads) in the following way:

Flagging – EVERYONE IN PROYECTO PRIMATES IN ECUADOR SHOULD FOLLOW THESE CONVENTIONS:


            In Ecuador, all trees get marked with a unique letter and number combination that includes information on the primate species using the tree, the year in which the tree was marked, and a sequential number. The first two digits of the tree number correspond to the first letter of the genus and species of the primate using the tree (“PA” for Pithecia aequatorialis, “CD” for Callicebus discolor, “AV” for Aotus vociferans,  “AB” for Ateles belzebuth, “LL” for Lagothrix lagotricha, “AS” for Alouatta seniculus). Then, the rest of the tree number consists of 2 digits corresponding to the year in which the tree was 1st used, a dash, and then a three digit sequential number (“001”, “002”, etc.). For example, the 1st tree marked in 2005 for owl monkeys was given the code “AV-05-001”, the second given “AV-05-002”, etc.).

            To mark the tree, write the tree number on PINK FLAGGING TAPE (please ONLY use pink!) and tie the tape around the tree, preferably UNDER (rather than around) any lianas hanging down or attached to the trunk. You should also write the date (in “mmm yy” format) and the initials “PP” for “Proyecto Primates” on the tape, and you should write on the tape “Sleep” if the tree is a sleeping tree or the Life Form of the feeding source if the tree is a feeding tree. Finally, you should write the tree number on a metal “write-on” or engravable tag that you then put on the tree with an aluminum nail. Go over the engraving with a black sharpie marker to make the number stand out at a distance. Do not hammer the nail all the way in; you should leave about an inch or so of shaft exposed on the nail to allow the tree to grow without swallowing the tag.

            Note that if you are marking a liana or other epiphytic plant that spans several trees, you need only tag the trunk that supports the bulk of the liana. In Ecuador, every new life form marked on a given trunk should be tagged as a different number. For example, say one day the titi monkeys eat fruit from a tree that you tag as “CD-05-104” and then they eat, a few days later, from a liana in the same tree crown, the liana should get a NEW TAG NUMBER. Similarly, if multiple species eat from the same life form, you should put up a tag and tape for each species. Thus, a given tree may be marked with 2 or more metal tags and pink flags. If you do record a new number for a previously marked tree, enter all the other tags separated by slashes in the “AKA” section of the marked tree table in the database. Also note in this section any numbers on the metal tags from other projects (e.g., "FT" tree numbers from Abigail Derby's work on howler monkeys; the numbers from Mark Mulligan and collaborators' HERB projects).

The following data will be noted in the Marked Trees section of your data book or on the Palm data logger:

AGAIN, IMPORTANT NOTE: Remember that when you note an angle, the angle should be AS IF YOU WERE STANDING AT THE MAPPED REFERENCE POINT. This means that if you take an angle from a tree back to a mapped point, you will have to invert the angle (e.g., an angle of 180 degrees read from where the monkeys are back to a reference point corresponds to an angle of 360 degrees from the reference point). Be very careful to not map a tree 180 degrees off from its true location!

 

Some Additional Useful Definitions for Tree Measurements, even if not prompted to enter them in the Database 

 

 

Feeding Bouts Tab:

            Additionally, for each marked tree, you will record feeding bout data in the Feeding Bouts section of your data book or the Palm data logger for all bouts greater than 5 “monkey minutes”. You need not worry about recording bout data for bouts lasting less than 5 minutes. For each feeding bout, note the time (to the nearest minute) at which the first feeder(s) entered the tree and began feeding (e.g., “10:45 BE AV-05-001” for “Begin Eat AV-05-001 at 10:45”) and when the last feeder(s) finished feeding (e.g., “11:15 EE AV-05-001” for “End Eat AV-05-001 at 11:15”). If you are following some animals and arrive at a food patch where animals are already feeding, score your arrival time as “BE Late” (“begin eat late”, check the box “Late?”). If you need to leave a tree before the last feeders have left (e.g., if your focal animal leaves the feeding patch while others are still eating), record the ending time as “EE Early” (“end eat early”, check the box “Early?”). If you miss a BE or EE time, score it as “Not Recorded”, but please be aware that this is a code of last resort, introduced to deal with when observers forgot to collect data that they were supposed to collect. With the BE and EE times, we can calculate a minimum number of minutes that the group spent feeding in a particular patch. If the animals stop feeding in a patch for more than five minutes, and they return to the same tree, record another set of BE and EE times. If the animals are feeding but you do not know from which tree you may record it as PA-05-XXX, meaning the monkeys are having a feeding bout but you cannot be sure on which tree, therefore you cannot mark it. Also, record the maximum number of individuals that you see feeding in the patch simultaneously (Max # Feeders).

            If the food item is one that you do not recognize or that you have not collected previously, MAKE A COLLECTION of both the leaves and the fruit of the plant (note in the Marked Trees section of your data book and in the database what collections you have made; you will enter specific information on the collection in the Plant Collections section of the database later). It is important to do these collections to allow us to identify the plants being eaten. To make collections, look carefully at the tree through binoculars to get a good idea of the appearance of the leaves and fruits and then look for these on the ground under the tree. Carry plastic bags with you at all times to shove these collections into until you get back to the lab. In Formosa, if you cannot identify the species, write down in the last page of the Avistaje yellow book the information on the tree so that the Botanist David Iriart can identify the tree when he goes to Formosa. In Ecuador, when you return to the lab with your collection, process the samples according to the Plant Voucher Specimens protocol.

Focal Samples Tab:

Ad Lib Data Tab: 

Enter in this tab data pertaining to observations that may have happened outside focals or that were not amenable to the systematic data collection that is done during a focal.

 

PHENOLOGY SUBFORM:

It is used to enter data on PHENOLOGICAL PATTERNS in the forest (see the Phenological Data Collection protocol).

 

BIOLOGICAL SAMPLES SUBFORM:

It is used to enter Data on any BIOLOGICAL SAMPLES you collect (see the protocols for Plant Voucher Specimens, Plant Nutrient Samples, Fecal Samples for DNA Extraction, Fecal Samples for Hormone Extraction, and Urine Samples for Hormonal Analysis).

 

MAPPING DATA SUBFORM:

It is used to enter MAPPING DATA for trails and transects around the site (see the Flagging, Trail Maintenance and Mapping protocol).