TITLE: Macrofauna Occidental Farms
AUTHORS: Carlos Alberto Zuniga Gonzalez, Conrado Ronaldo Quiroz Medina, Alvaro Jose Caballero Hernandez

[DESCRIPTION]
The protocol was divided into two phases, the first was the collection of the sample, and the second was the classification, coding, and storage of the extracted macrofauna populations. Subsequently, the indices of diversity and species richness, relative abundance, by functional groups were estimated. The sampling consisted of the extraction of 40 soil samples at a depth of 0-20 cm and 40 samples of litter on the soil surface of the 8 banana farms sampled and distributed in the municipalities of León and Posoltega, for a total of 80 samples collected, coded, stored and transported to the Entomology Laboratory of the Department of Agroecology, School of Agrarian and Veterinary Sciences of the National Autonomous University of Nicaragua, Leon.

[BEFORE_START]
This protocol is part of the research untitled Diversity of Functional Edaphic Macrofauna in Musa acuminata x Musa balbisiana (AAB) Agroecosystems, the sampling design was random and descriptive, it was divided into two phases, the first was the collection of the sample, and the second was the classification, coding, and storage of the extracted macrofauna populations. Subsequently, the indices of diversity and species richness, relative abundance, by functional groups were estimated. 

[STEPS]
12. Phase 1: Field: It includes steps 1, 2, 3, 4, 5: Following the methodology modified by Rousseaua in the cocoa plantations in Costa Rica and by Laurente in the Quenzalgual systems in agroforestry systems. (Rousseaua, Deheuvelsb, Rodriguez-Arias, & Somarriba, 2012; Rousseaua, Fonte, Téllez, van der Hoekc, & Lavelle, 2013)
4. First Step: Delimitation of the Sampling Area
Upon arrival at each farm, an area of 0.7 hectares established with banana cultivation was located and an area of 1000 m² (50m x 20 m) was delimited with a tape measure. This study plot was divided into 5 subplots each one of 10 m² x 20 m². Where a wooden frame with the dimensions of 20 centimeters long and 20 centimeters wide was used to mark the extraction point.
12. Second Step: Litter removal process at ground level and soil from 0-20 cm
   Already put the wooden frame (20 cmx20cm) on the ground. We use a shovel to border the outside of the wooden frame to delimit the litter block and the soil column from 0 to 20 cm to obtain an approximate weight of 1 kilogram of soil; divided into two successive strata (leaf litter, 0-20 cm) each side was surrounded with plastic mesh so that soil fauna did not escape with the least possible disturbance.
12. Third Step: Sieving of the litter-soil and separation of macrofauna
  The sieving (20 mesh x inch² sieve) of the litter and the soil volume of 0-20 cm was carried out with great care, manually separating the soil, clods, roots, twigs, and stones from the macrofauna found.
12. Fourth Step: Manual collection of each insect and placement in airtight jars
After separating the litter and soil from the macrofauna found in the sampling area. Each insect is taken and placed inside an airtight plastic jar with dimensions of 500 cubic centimeters in volume, labeled, coded, and preserved with 70% alcohol, and the worms are preserved with 4% formalin.
12. Fifth Step: Transfer of samples to the laboratory
  The labeled and coded containers were transferred to the Entomology Laboratory of the Agroecology Department of UNAN-León, for proper identification. Protecting samples from the sun.
4.1. Phase 2: Laboratory: Includes step 6: Scientific Reference: The methodology of these Invertebrate Identification Manuals (Maes, 1998; King, 1984; Andrews & Caballero, 1990) was followed.
4.2. Step Six: Identification of the Macrofauna in the Laboratory
The collected individuals were quantified and identified by order, family, and gender with the help of invertebrate identification manuals in the Entomology Laboratory of the Department of Agroecology. The organisms were placed in a Petri dish and then observed with the help of a 4x binocular stereoscope, where the particular structures of each species were detailed. The macrofauna includes all those organisms whose length is greater than 4 mm. Each individual was separated in a bottle according to its origin, sample number, and biological classification.
12. Step Seven: Uses of macrofauna richness and diversity indices
The diversity and richness of the species present in this study were analyzed using the following indices reported by the authors (Moreno, 2001; Rousseaua, Deheuvelsb, Rodriguez-Arias, & Somarriba, 2012; Zerbino, 2005):
Richness (number of species), DAFGA (RRID:SCR_003319) :
Relative abundance (Density number of individuals 1 m² for each species):
Functional Groups (Density number of individuals 1 m² for each species):
Shannon-Wiener Index (SW):
Margalef Diversity Index
Proportional abundance indices
Dominance indices:
Simpson index:
Equity indices:
Pielou index:
12. Phase 3: Producer Consultation Yield of fruits per apple (number of fruits/ha)
Eighth Step: Yield of fruits per hectarea
The total number of fingers per ha was recorded with the help of the producer who provided us with the data according to the records kept on the farm.