USER INSTRUCTIONS

User Instructions can be downloaded as a pdf file >>>   notes.pdf


EPG-Calc is designed to analyze raw data files provided by the dedicated softwares Probe or Stylet+ for Windows operating systems and automatically calculate 142 parameters.
EPG-Calc is accessed at http://www2.sophia.inra.fr/ID/epg.php.



page1

Screen 1 allows the user to connect and download user instructions and the list of calculated variables in Portable Document Format (".pdf") files.



page2

Screen 2 Information necessary for data processing have to be filled in.
- The option "aphid species with expected waveforms sequence" is checked by default. If the data have been collected on non sheath-feeding hemipterans exhibiting non stereotypical behavioral sequences the option "species with non conventional characteristics and sequence of EPG waveforms" has to be selected.
- The analysis can be performed from "the start of the recording time" which is the commonly used mode. However, because time to initiate the first probe (stylet insertion within plant tissue) could be highly variable, the analysis can be realized "from the start of the first C phase" which allows to compare identical probing durations.
- "Experimental time in seconds" is the time from the initial recording time that will be used to calculate parameters (28,800 sec i.e. 8 h by default). For instance a 8 h recording experiment is too large when only intracellular probes (potential drops) are studied. Once validate by uploading the first file on the server, this variable is set permanently for the user session.
- As EPG-Calc allows to upload files from different experimental situations independently of their uploading order, each file has to be identified by its "Experimental class name". The file to upload is directly chosen by clicking the "Browse..." button, no modification of the raw data files provided by the analysis software (Probe or Stylet+) being necessary. If not specified in the data file, EPG-Calc automatically marks the start and/or the end of each recording. Raw data files, i.e. Probe or Stylet+ ".ana" files (1 file per individual), can be uploaded one by one on the server. To speed up the uploading process, all the files from ".ana" format of a same experimental situation can be uploaded once in a ".zip" archive file (".7z" or ".rar" file format are not accepted). ZIP archives are managed by the PclZip 2.8.2 PHP library (http://www.phpconcept.net/pclzip). By clicking "Submit" button, the different filled fields are checked and the file (only ".ana" and ".zip" format files are allowed) is uploaded.
If the option "aphid species with expected waveforms sequence" is checked the uploaded EPG data file is automatically checked for errors in the expected sequence of waveforms exhibited by phloemophagous aphids such as two identical waveform codes in successive rows, a lower time than that of the preceding row, or an ingestion (E2) phase not preceded by a salivation bout (E1).



error

Only coding and duplication errors are checked when the option "species with non conventional characteristics and sequence of EPG waveforms" is selected. In such a case, an error message is displayed identifying the row and the waveform code or time column in a specific file where an error occurs, and the corrupted file is deleted from the server. In the case of a ."zip" file note that the whole archive is deleted and has to be uploaded again.



page3

Screen 3 is accessed if the raw data file is successfully checked ("file (n° XXX) successfully analyzed") and 2 buttons "Calculate EPG parameters" and/or "Upload another file" are displayed. It is noteworthy that at least 2 files per experimental class are needed to calculate EPG parameters. Clicking "Upload another file" redirects to Screen 2.
By selecting "Calculate EPG parameters", average, standard deviation, standard error of the mean and the number of individuals used to determine each parameter statistics are calculated for each experimental class and the session is stopped.



page4

Screen 4 allows the user to choose the file format, database-compatible CSV text by default or Microsoft Excel® XLS, which will be sent as a mail attachment. XLS files are generated using php_writeexcel which is a part of Spreadsheet::WriteExcel Perl package to PHP (http://www.bettina-attack.de/jonny/view.php/projects/php_writeexcel/).



page5

Screen 5 Clicking "Ok" ends the session.



NOTES


i) EPG parameters of individuals and EPG statistics are sent as:
   - 2 database compatible ".csv" files:
      DATA_*****.csv: EPG parameters of individuals (1 individual = 1 Probe/Stylet file)
      STAT_*****.csv: descriptive statistics of experimental classes (screenshot below)
   - or 1 Microsoft Excel® ".xls" file with 2 worksheets.

screen

ii) If a phase (xxx) is missing only the following variables are evaluated:
   t>1xxxrec as the recording time
   t>1xxx as the time from the 1st Pr to the end of the recording
   the other variables appear as "-" in the DATA_***.csv file or in the DATA worksheet of the MS Excel workbook.

iii) For each file to upload the maximum size is 1,000,000 bytes

iv) When using ".zip" file, all the raw data files (".ana") have to be at the root of the archive