Revista Pastos y forrajes vol41n4 2018

Vol. 41, No. 4, October-December 2018 / NRS 0099
ISSN 0864-0394 (printed version) / ISSN 2078-8452 (online version)
Quarterly journal. Official organ of the Ministry of Higher Education for pastures and forages | 1978
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TOPICS
• Introduction, evaluation and dissemination of
plant genetic resources related to the farming
sector.
• Agroecological management of production
systems.
• Sustainable livestock production.
• Conservation of forages and agroindustrial
byproducts for animal feeding.
• Agroforestry for animal and agricultural
production.
• Integrated food and energy production
systems in rural areas.
• Utilization of alternative medicine in tropical
farming systems.
• Adaptation to and mitigation of the climate
change in farming ecosystems.
• Economic, managerial and social aspects of
farming production.
• Extension, agricultural innovation and
technology transference.
• Rural and local development.
ESTACIÓN EXPERIMENTAL DE PASTOS Y FORRAJES INDIO HATUEY
EDITORIAL COUNCIL
Editor-in-Chief | Dra. Tania Sánchez Santana
Assistant Editor | M.Sc. Nayda Armengol López
Editor-Agricultural Sciences | Dra. Marta Hernández Chávez
Editor-Veterinary Sciences | Dr. Javier Arece García
EDITORIAL COMMITTEE
Dr.C. Jesús Suárez Hernández | Dra.C. Maybe Campos Gómez
Dra.C. Marlen Navarro Boulandier | Dra.C. Maykelis Díaz Solares
Dr.C. Jesús M. Iglesias Gómez | Dr.C. Hilda B. Wencomo Cárdenas
Dr.C. Anesio R. Mesa Sardiñas | Dr.C. Luis A. Hernández Olivera
Dra.C. Hilda C. Machado Martínez | Dr.C. Osmel Alonso Amaro
Dr.C. Giraldo J. Martín Martín | Dra.C. Odalys C. Toral Pérez
Dr.C. Luis Lamela López | M.Sc. Onel López Vigoa
Dra.C. Mildrey Soca Pérez | M.Sc. Milagros de la C. Milera Rodríguez
Dr.C. Félix Ojeda García | M.Sc. Yolai Noda Leyva
Dr.C. Gertrudis Pentón Fernández | M.Sc. Juan C. Lezcano Fleires
SCIENTIFIC COMMITTEE
Dra. Sonia Jardines González | Universidad de Matanzas, Cuba
Dra. Angela Borroto Pérez | UNIVERSIDAD DE CIEGO DE ÁVILA, Cuba
Dr. Aníbal E. Fernández Mayer | Instituto Nacional de Tecnología
Agropecuaria, Argentina
Dr. Argemiro Sanavria | Universidad Federal Rural de Rio de Janeiro, Brasil
Dr. Tyrone J. Clavero Cepeda | Universidad de Zulia, Venezuela
Dr. José M. Palma García | Universidad de Colima, México
Dr. Oscar Romero Cruz | Universidad de Granma, Cuba
Dr. Carlos J. Bécquer Granados | Estación Experimental de Pastos y Forrajes
de Sancti SpÍritus, Cuba
Dr. Rodobaldo Ortíz Pérez | Instituto NACIONAL de Ciencias agrícolas, CUBA
Dr. Pedro C. Martín Méndez | Instituto de Ciencia Animal, Cuba
Dr. Pedro P. del Pozo Rodríguez | Universidad Agraria de La Habana, Cuba
Dr. Redimio Pedraza Olivera | Universidad de Camagüey, Cuba
Dr. Rafael S. Herrera García | Instituto de Ciencia Animal, Cuba
Dr. Pedro José González Cañizares | Instituto Nacional de ciencias agrícolas, CUBA
Dr. Ángel Arturo Santana Pérez | Universidad de Granma, Cuba
SUPPORT COMMITTEE
Editing and correction
M.Sc. Alicia Ojeda González
Design and editing
Dailys Rubido González
Miresleidys Rodríguez Rizo
Translation
B.A. Nidia Amador Domínguez
Cover design
B.A. Israel de Jesús Zaldívar Pedroso

Vol. 41, No. 4, October-December 2018
Revista Trimestral. Órgano oficial del Ministerio de Educación Superior para el área de los pastos y forrajes
Quarterly journal. Official organ of the Ministry of Higher Education for pastures and forages
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CONTENT
| analysis and comments |
Innovation processes in local food and energy production in Cuban
municipalities
Jesús Suárez-Hernández, Julio Ramiro Quevedo-Benkí, Maikel Rodolfo Hernández-
Aguilera, Abel Peña-Alfonso and Guillermo González-Telles..................221
| scientific paper |
Effect of hydric stress on the germination of seeds from Sorghum
bicolor (L.) Moench cv. UDG-110
Yunel Pérez-Hernández, Marlen Navarro-Boulandier, Leannys Rojas-Sánchez,
Leticia Fuentes-Alfonso and Maryla Sosa-del Castillo..............................226
Population performance of a collection of Urochloa brizantha (Hochst.
ex A. Rich.) R.D. Webster in association with Stylosanthes guianensis
CIAT-184 (Aubl.) Sw
Yuseika Olivera-Castro, Pedro Pablo del Pozo-Rodríguez, Lisset Castañeda-
Pimienta and Juan Francisco Ramírez-Pedroso...........................................236
Performance of the edaphic macrofauna in animal husbandry systems, in
a productive entity of the Yaguajay municipality, Cuba
Wendy Mercedes Ramírez-Suárez, Marta Beatriz Hernández-Chávez, Alexis
Abilio Zurita-Rodríguezand Marlen Navarro-Boulandier.........................241
Milk production and bromatological quality and costs of supplementation
with Tithonia diversifolia (Hemsl.) A. Gray, in Jersey cows
Luis Mauricio Arias-Gamboa, Andrés Alpízar-Naranjo, Miguel Ángel Castillo-
Umaña, María Isabel Camacho-Cascante, Victoria Arronis-Díazand José
Enrique Padilla-Fallas.....................................................................................248
Silvopastoral system with Tithonia diversifolia (Hemsl.) A. Gray: effect
on the rumen microbial population of cows
Juana L. Galindo-Blanco, Idalmis Rodríguez-García, Niurca González-Ibarra,
Roberto García-López and Magaly Herrera-Villafranca...........................254
Evaluation of a system of selective antiparasitic treatments in Pelibuey
sheep during mating and pregnancy
Javier Arece-García, Yoel López-Leyva, Roberto González-Garduño, Ramón
Luck Montero and Leticia de la Caridad Carballo-Silverio.......................261
Meals from woody forage plants and palm fruit in the diet of broilers
Emiro Rafael Canchila-Asencio, Julio Cesar Rodríguez-Galvis, Rodolfo
Corredor-Barrios and Ubaldo Navarro-Contreras......................................267
Contribution of innovation to local development management in a
municipality of Matanzas
Taymer Miranda-Tortoló, Hilda Machado-Martínez, Juan Carlos Lezcano-Fleires,
Antonio Suset-Pérez, Katerine Oropesa-Casanova, Frank David Tirado-
García, Luis Lamela-López and Iván Lenin Montejo-Sierra.....................272
Evaluation of mixtures of Jatropha curcas (L.) biodiesel in diesel engine banks
Indira Tobío-Pérez, Eliezer Ahmed Melo-Espinosa, Jesús Suárez-Hernández,
José Ángel Sotolongo-Pérez and Ramón Piloto-Rodríguez.......................279

Pastos y Forrajes, Vol. 41, No. 4, October-December, 221-225, 2018 / Innovation processes in local food and energy production 221
Analysis and Comments
Innovation processes in local food and energy production in Cuban
municipalities
Jesús Suárez-Hernández1
, Julio Ramiro Quevedo-Benkí1
, Maikel Rodolfo Hernández-Aguilera2
,
Abel Peña-Alfonso3
and Guillermo González-Telles4
1
Estación Experimental de Pastos y Forrajes Indio Hatuey, Universidad de Matanzas, Ministerio de Educación Superior
Central España Republicana, CP 44280, Matanzas, Cuba
2
Unidad de Extensión, Investigación y Capacitación Agropecuaria, Velazco, Holguín, Cuba
3
Centro de Desarrollo Local, Gobierno Municipal de Manatí, Las Tunas, Cuba
4
Filial Universitaria de Martí (Universidad de Matanzas), Martí, Matanzas, Cuba
E-mail: jesus.suarez@ihatuey.cu
Abstract
The objective of the work is to make an analysis of the diverse innovation processes in the local integrated food and
energy production in Cuban rural areas in the context of the Biomas-Cuba project1
. For the identification and evaluation
of the local innovation process in the integrated food and energy production (IFEP), three questions were established:
1) what innovation processes have been developed or introduced?, 2) What has the link consisted in?, and 3) Which have
been the key learnings? The IFEP is promoted for the development of agroenergetic farms, which has been implemented
in private farms, cooperatives and state farms, in 22 municipalities of six Cuban provinces. They concentrate two types
of innovation: i) sowing and agronomic management of Jatropha curcas L. for biodiesel production, and ii) integration
of biodigesters in animal and plant production farms. In addition, 176 biodigesters were designed and constructed,
mainly fixed-dome ones, and two anaerobic lagoons covered with synthetic geomembrane. In the organizational context
there was a link among research centers, municipal universities, governments and local state entities, and professional
and farmer organizations. It is concluded that the promotion of local innovation processes in agroenergy, in Cuban
municipalities, contributes to a new strategic approach for the IFEP from biomass in rural areas, in the framework of
local innovation systems, with high participation of actors, open innovation processes and a science-productive sector-
government link.
Keywords: bioenergy, extension, food production
1
Biomas-Cuba: International project led by the Pastures and Forages Research Station Indio Hatuey (EEPFIH) and funded by the
Swiss Development and Cooperation Agency (SDC).
Introduction
At global scale there is a challenge: how to
make agroenergy, food security and environmental
protection coexist?, in the presence of climate
changes, environmental degradation, food crisis
and the «biofuels vs. food» contradiction, generated
by a senseless policy to obtain the former in large
land extensions which were previously dedicated
to produce food, something morally rejectable.
Nevertheless, biofuels are an ecological alternative
to substitute fossil biofuels, in the rural context, and
reduce the emissions of greenhouse gases (GHG).
This is enhanced in integrated agricultural systems,
in which biofuels and food can be produced (Suárez
et al., 2011).
This approach to produce, in an integrated
way, foodstuffs and biofuels (liquid, gaseous and
solid), demands to develop learning and innovation
processes, create a remarkable absorptive capacity
in the different involved actors and promote
innovation systems and interrelations among the
governments, universities, research centers and
the productive sector, which constitute important
catalysts for a successful implementation of such
integrated approach.
In this regard, there are in literature diverse
antecedents which approach: learning and cons-
truction of technological and innovation capacities
(Ramirez et al., 2014), absorptive capacity (Warda and
Johansson, 2014; Castellacci and Natera, 2015a),
open innovation (West and Lakhani, 2008; West et
al., 2014), innovation systems (Castellacci and Na-
tera, 2015b; Wu et al., 2017), local innovation sys-
tems (Saldaña, 2014; Guercini and Runfola, 2015),
the university-enterprise link (Motoyama, 2014)
and the Triple Helix –the relation among the academy,

222 Pastos y Forrajes, Vol. 41, No. 4, October-December, 221-225, 2018 / Jesús Suárez-Hernández
enterprises and Government– (Deakin and Ley-
desdorff, 2014; Gebhardt, 2015; Leydesdorff et al.,
2017).
These approaches configure the national, re-
gional and local environment for innovation, for
which the literature on the topic has structured a
solid theoretical framework for understanding such
processes. However, the large majority of the works
have been focused on the experience of manufac-
turing and service enterprises, and few efforts are
still made to understand these issues in the agricul-
tural sector and, specifically, in bioenergy.
For such reason, the objective of the work
was to make an analysis of the diverse innovation
processes in the local integrated food and energy
production in Cuban rural areas in the context of
the Biomas-Cuba project.
Integrated food and energy production in the
rural context
The Biomas-Cuba project started in 2009, in a
context in which considerable volumes of foodstuffs
and fuels were imported, and the use of renewable
energy sources (RES) at that time was not a high
priority for the Government, unlike food production.
Since its conception, the decision was made to focus
it on three topics: biodiesel, biogas and gasification,
and their production and utilization in the framework
of the agricultural systems to contribute to food pro-
duction, to cost reduction and to the improvement of
rural life and environment quality.
In this sense, the project contributed in the in-
tegrated production of food and Jatropha curcas
L. biodiesel; biogas and biofertilizer production
from the effluents of biodigesters; biomass gasifi-
cation to generate energy; evaluation of integrated
systems to produce food and energy in Cuba; and
the generated economic, social and environmental
impact, with a strong link between the communi-
ties and decision-makers, as well as a remarkable
science-productive sector-government link (Suárez
and Martin, 2012). These results were obtained in
six Cuban provinces and contributed to improve
the quality of life, through the integrated food and
energy production (IFEP), in harmony with the en-
vironment.
The importance of this experience lies on the
promotion of local innovation systems in bioenergy,
based on open innovation processes among multi-
ple actors (researchers, farmers, decision-makers,
state institutions and communities), in which a new
concept emerges: the agroenergetic farm2
, new ap-
propriate technologies and applications are dissemi-
nated in Cuba, national and international networks
are promoted, and incidence is made on public poli-
cies, all this supported by the formulation and im-
plementation of local IFEP strategies.
For the identification and evaluation of the inte-
grated food and energy production in Cuba, in the
rural context, and for the characterization of the
link among the productive sector, the researchers,
professors and decision-makers of the government,
three questions were established for the analysis of
innovation processes:
1. What innovation processes have been developed
or introduced?,
2. What has the link consisted in?, and
3. Which have been the key learnings?
Innovation processes developed or introduced
in the rural context
The IFEP was promoted through the imple-
mentation of agroenergetic farms, located in private
farms, cooperatives and state farms, in 22 munici-
palities of the provinces Guantánamo, Holguín,
Granma, Las Tunas, Sancti Spíritus and Matanzas
(40 % of the Cuban provinces). In these systems
mainly two innovation types are concentrated:
i) sowing and agronomic management of food
crops in association with J. curcas, an adequate
shrub for biodiesel production from the seeds of its
fruits, and ii) integration of biodigesters in animal
and plant production farms.
In that sense, the first model for biodiesel pro-
duction emerged from the evaluation of a germplasm
of non-edible oil plants, such as J. curcas, Ricinus
communis L. and Aleuritis trisperma, introduced
in the country or collected in rural areas of Cuba,
which were established in germplasm banks of three
provinces (Machado et al., 2012). This evaluation
was complemented by the one carried out along with
several farmers in their own farms, which allowed
to identify promising materials, mainly of J. curcas,
for their seed and oil yields. Taking into considera-
tion the aspects reported about J. curcas and the
physical-chemical characteristics of its oil, it was
2
Defined as «the productive exploitation where technologies and innovations are developed, improved and evaluated to produce,
in an integrated way, food and energy (Suárez et al., 2011).

identified as the most appropriate plant for producing
biodiesel in Cuba (Sotolongo et al., 2012).
Planting combinations of J. curcas were evalua-
ted and 21 agricultural crops were intercropped
between the rows, among which beans, soybean,
peanut, corn, cassava, sorghum and rice stood out
with the best yields, with the application of survival
irrigation and fertilization with biofertilizers (Soto-
longo et al., 2012). This experience of intercropping
in the J. curcas rows proved that it is possible to
increase the productivity per hectare with spatial
arrangements that allow to produce biodiesel and
crops for human consumption.
On the other hand, the reforestation with J. cur-
cas was performed in 74 % of the soils not used
for agriculture, with diverse degradation conditions
–salinity, erosion, low fertility–, in high-fragility
areas with environmental affectations and, in some
cases, in degraded basins. This generated environ-
mental impacts, because it has been observed that
such plantations sequester annually 1 956 000 t of
carbon dioxide (CO2
), an important GHG; J. curcas,
for example, captures 6 kg of CO2
/year/tree (Suárez
and Martín, 2012).
The second innovation model (biogas and bio-
fertilizer) promotes the construction of biodigesters
for the anaerobic treatments of pig and/or cattle ex-
creta, which generate biogas as energy porter and
high-quality biofertilizers. In this context 176 bio-
digesters were designed and constructed, mainly
fixed-dome ones (up to 90 m3
of capacity), besides
polyethylene tubular and two anaerobic lagoons
covered with high-density synthetic geomembrane3
of 400 and 5 000 m3
. These facilities generate re-
markable productions, of biogas4
(which is used in
food and feed cooking, refrigeration, generation of
electricity and irrigation5
), as well as biofertilizers6
,
from the biodigester effluents, aimed at the ame-
lioration of degraded soils (Suárez, 2017). In this
innovation process the link among several research
centers, pig production enterprises and farmers
has been essential, which has generated synergies
and diverse improvements in the technologies, de-
signs and components of the different biodigester
systems.
Another innovation of impact on the quality of
life of the inhabitants is the creation of four biogas
supply networks, fed by biodigesters, which benefit
53 houses and 272 persons in the Cabaiguán munici-
pality (in the central region of the country), which
constitute the first rural communities in Cuba with
a gas supply network in food cooking and other
usages, annually saving 77,2 MWh of electricity.
The gasification of biomass for energy genera-
tion, more efficient process than the traditional
combustion, as firewood or charcoal, is also the ob-
ject of analysis. Two gasifiers and their generators
were installed, with capacity of 20 and 40 kW of
power, at the Pastures and Forages Research Station
Indio Hatuey (EEPFIH), in Matanzas province, and
in a sawmill, in Santiago de Cuba province, ope-
rated with Dichrostachys cinerea (L.) Wight &
Arn. branches and trunks and with wastes from the
pruning of animal husbandry agroforestry systems
and from the processing of timber (bark), respec-
tively (Cepero et al., 2012).
Economic and environmental appraisals were
made in order to evaluate the IFEP in different
productive scenarios of 15 municipalities. An in-
crease of local food production (vegetables, fruits,
milk, meat and eggs) was found, influenced by the
above-mentioned innovations, from 1,6 to 27,3 mil-
lion Cuban pesos (CUP), and the productive items
were remarkably diversified. Likewise, between
2013 and 2016 88,7 million CUP were generated,
due to the substitution of food, fuel and fertilizer
imports for a value of 5,9 million USD –with-
out considering the savings in maritime freights
(Suárez, 2017).
In the improvement of the farmer families’
quality of life, the following indicators stand out:
the creation of 372 new direct jobs, with a mean
monthly salary higher than the average salary of the
involved provinces, from which 28 % are occupied
by women, favoring the family economy.
A positive environmental impact has also
been generated associated to the reforestation with
335 000 trees, which favors carbon sequestration;
substitution of fossil fuels and decrease of CO2
and
SO2
emissions due to the management of the pig and
cattle excreta; production of biofertilizers, which
contributes to improve soil fertility, among others.
3
Adequate technology for large waste volumes, solving the limitations of uncovered anaerobic lagoons, which emit methane and
unpleasant odors.
4
At present 1 145 317 m3
of biogas/year are generated (equivalent to 3 460 barrels of oil).
5
A total of 12 841 people who inhabit rural zones of Cuba is benefitted.
6
At present, 90 423 t per year are produced; this has allowed to improve 3 874 ha of degraded soils.

224 Pastos y Forrajes, Vol. 41, No. 4, October-December, 221-225, 2018 / Jesús Suárez-Hernández
A key catalyst of this entire process has been
the formulation and implementation of local IFEP
strategies in six municipalities, which are integrat-
ed to their Local Development strategy, which has
allowed to provide their governments with a stra-
tegic management tool to promote, in the synergy
framework, food security, utilization of RES, waste
treatment and utilization, land reclamation, GHG
reduction and mitigation and adaptation to the cli-
mate change.
Importance of the science-productive sector-
government link in the innovation processes
These results were obtained through the pro-
motion of intense open innovation processes by re-
searchers and farmers and between them, to which
local decision-makers from the governments and
state entities were incorporated, as well as the mu-
nicipal university centers and campuses. This al-
lowed to create a kind of local innovation systems,
in which the participation and inclusion of every-
one, individual and organizational learning, and the
creation of absorptive and local innovation capaci-
ties, are permanent elements.
In the participating municipalities, this link be-
came closer among the Local Operational Commit-
tee, Council of Municipal Administration (CMA),
Municipal Delegation of Agriculture; municipal
directions of Physical Planning, Economy and
Planning, Hydraulic Resources and of the National
Association of Small Farmers (ANAP, for its ini-
tials in Spanish), the Municipal University Center
and other projects that act in these territories, with
which important synergies are achieved. In this
sense, each semester in the CMAs –the local execu-
tive power– the implementation of the above-ap-
proached Local IFEP Strategies, which exist in six
municipalities, is evaluated; while once per year
this evaluation is made by the Municipal Assem-
blies of People’s Power –the local legislative power.
The organizational context for the implemen-
tation of these innovation processes is shown in a
permanent link among research centers, municipal
university campuses, producers –with emphasis on
farmer men and women–, governments and local
state entities and professional and farmer organiza-
tions, which more than a Triple Helix is a Four-He-
lix –enlarged with the civil society–, which has
allowed to have incidence on local public policies.
Likewise, the local actors interact periodically
with national and sectorial decision-makers, linked
to the Ministries of Energy and Mines (Minem),
Agriculture (Minag), Higher Education (MES),
Industries (Mindus) and Science, Technology and
Environment (Citma), which contributes to have in-
cidence not only on local public policies, but also
at ministry and national level. As examples of this
interaction the following stand out: i) the Directions
of Renewable energy and of Energy Policy of the
Minem, ii) several enterprises from the metal-me-
chanic, chemical and appliances industries to manu-
facture equipment and components of the biogas
and biodiesel systems, and iii) the Entrepreneurial
Agricultural, Agroforestry, Animal Husbandry and
Labiofam groups, to train their managers and spe-
cialists, as well as to implement diverse technolo-
gies for bioenergy.
Key learnings in the IFEP
Concerning the lessons learned or under cons-
truction, the following stand out:
• The thematic approach focused on the IFEP, with
the application of the concept of agroenergetic
farm, which has allowed to create in diverse ac-
tors a more integrated and systemic conception.
• A wide work in the network among all the ac-
tors and synergies at local, territorial and na-
tional scale, supported on the participation in
multi-institutional and multi-actor platforms, as
well as a link among the academic sector and the
farmers and decision-makers.
• Intense processes of local agricultural innova-
tion, in the framework of an open innovation
model aimed at the achievement of practical re-
sults, in which technologies and innovations are
developed and improved with wide participation
of the beneficiary, for the sustainability of the
actions.
• Direct links with the farmers and their families,
which has allowed to accompany them in the de-
velopment of sustainable production processes
with community participation.
• The promotion of synergies with other interna-
tional projects and with institutions (ministries,
local governments, enterprises, technical and
producers’ organizations, and farmers).
• The permanent process of systematization and
socialization of results, experiences, good prac-
tices, technologies and designs, among others,
aimed at direct beneficiaries and project mana-
gers, policy decision-makers and at the scienti-
fic and academic sector, at local, provincial and
national scale.

• Active contribution to the formulation of the na-
tional biogas and biodiesel programs.
Conclusions
The promotion of local innovation processes in
agroenergy, in Cuban municipalities, contributes to
a new strategic approach for the IFEP from biomass
in rural areas, in the framework of local innovation
systems, with high participation of actors, open
innovation processes and a science-productive sec-
tor-government link.
Acknowledgements
The authors thank the international project
Biomas-Cuba, funded by the Swiss Development
and Cooperation Agency (SDC), which contributed
with the funding for the implementation of
technologies and innovations linked to food and
energy production in the Cuban rural context.
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Castellacci, F. & Natera, J. M. The convergence para-
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Received: January 20, 2018
Accepted: November 16, 2018

226 Pastos y Forrajes, Vol. 41, No. 4, October-December, 226-235, 2018 / Yunel Pérez-Hernández
Scientific Paper
Effect of hydric stress on the germination of seeds from Sorghum
bicolor (L.) Moench cv. UDG-110
Yunel Pérez-Hernández1
, Marlen Navarro-Boulandier2
, Leannys Rojas-Sánchez1
, Leticia
Fuentes-Alfonso1
and Maryla Sosa-del Castillo1
1
Centro de Estudios Biotecnológicos, Facultad de Ciencias Agropecuarias, Universidad de Matanzas, Cuba
2
E-mail: yunelph@gmail.com
Abstract
The objective of this work was to evaluate morphological and biochemical indicators in Sorghum bicolor (L.)
Moench cv. UDG-110, during the germination process under conditions of hydric stress induced by polyethylene
glycol-6000. The seeds were sown on Petri dishes with different concentrations of polyethylene glycol-6000 (0, 3, 6, 9,
12, 15, 18 and 21 %) and were placed in a growth chamber during eight days. The following indicators were evaluated:
germination percentage, germination value, length of the root and the aerial part, α-amylase activity and contents
of proteins, reducing sugars and soluble phenols. A completely randomized design was used with four replicas per
treatment. Polyethylene glycol-6000 affected the germination percentage and the morphophysiological traits, such
as length of the roots and aerial parts. The α-amylase activity increased in the variants with presence of the osmotic
agent, with values higher than 9 %. In the low and intermediate concentrations of polyethylene glycol-6000 the highest
contents of reducing sugars and total soluble proteins in the root and the aerial part were observed, respectively. The
concentration of soluble phenols in the aerial part reached high values between 15 and 18 %, which can be related with
an antioxidant defense mechanism to face the consequences of the oxidative stress generated under diverse conditions
of abiotic stresses. Polyethylene glycol-6000 affected the germination process of S. bicolor cv. UDG-110, although
there was germination with 21 %, showing the presence of drought tolerance mechanisms, such as the production of
osmotically active compounds and the synthesis of antioxidant substances.
Keywords: sorghum, reducing sugars, phenolic compounds, drought
Introduction
Drought is one of the most important
environmental stresses that affect crop growth,
development and productivity. The understanding
of tolerance mechanisms of plants to this stress
constitutes one of the most important challenges
for agricultural researchers and for plant breeding
(Mujtaba et al., 2016). This situation is worsened
if the consequences of the climate change are
considered, which aggravates the abiotic stresses
at global scale by increasing the irregularity of
meteorological events (Jain and Saxena, 2016). For
such reasons, it is necessary to develop strategies
that allow crop adaptation to specific environments
and to identify at early ages those that can maintain
high yields under water deficit conditions (Khaton
et al., 2016).
Sorghum [(Sorghum bicolor (L.) Moench)] is
a highly demanded crop at global scale because of
its usage in human and animal feeding, as well as
its high potential for renewable energy production
(Rezende et al., 2017). This species was catalogued
as moderately tolerant to hydric stress and although
water scarcity can exert an adverse effect on the
germination process, many sorghum cultivars
adapt well to semiarid conditions (Achón-Forno et
al., 2014).
Germination and the first stages of plant growth
constitute the most vulnerable moments within the
life cycle of plants. Good germination can contribute
remarkably with the transit of the plant through the
different phenological stages (Channaoui et al.,
2017). In this sense works have been conducted
about the response of S. bicolor to hydric and saline
stress during the germination process, in order to
understand the tolerance mechanisms that occur
in this species (Tsago et al., 2013). These studies
are fundamental for establishing later successful
breeding programs in this crop (Khan et al.,
2015). The objective of this wok was to evaluate
the effect of hydric stress induced by polyethylene
glycol-6000 on the germination of seeds from
S. bicolor cv. UDG-110.

Pastos y Forrajes, Vol. 41, No. 4, October-December, 226-235, 2018 / Hydric stress on the germination of seeds from Sorghum bicolor 227
Materials and Methods
Seeds from sorghum cv. UDG-110 were used,
supplied by the Pastures and Forages Research
Station Indio Hatuey (eePFIH), of Matanzas
province, Cuba: while the studies were conducted
in the laboratory of Biotechnology of the School
of Agricultural Sciences, University of Matanzas,
Cuba.
The essays were developed according to a
completely randomized design. In this sense,
the seeds were placed on Petri dishes of 5 cm
diameter on filter paper moisturized with different
concentrations of polyethylene glycol-6000 (0, 3,
6, 9, 12, 15, 18 and 21 %), in a proportion of three
times the weight of the substrate which constituted
the treatments. Twenty five seeds were placed
per Petri dish and four replicas were made per
treatment (ISTA, 2010). The germination process
was evaluated daily during eight days and the
results were expressed in percentage of germinated
seeds. For the biochemical analyses five samples
were taken per treatment; while for the evaluation
of the morphological and physiological indicators
10 seedlings were analyzed.
Measurements
Germinationvalue.Thequantityofgerminated,
non-germinated and rotten seeds was determined
daily during the experiment. With the obtained
data the germination value (GV) was calculated,
according to the formula proposed by Djavanshir
and Pourbe (1976).
Biochemical indicators. The root and aerial part
of the germinated seedlings were cold macerated with
buffer solution of sodium phosphate 50 mmo/L, pH
7,0 and in a proportion 1:3 (w/v). The homogenate
was centrifuged at 12 000 rpm and the supernatant
was collected, which was preserved at -20 ºC until
the moment of the determinations. The biochemical
indicators were carried out in all the treatments,
except in the highest concentration of polyethylene
glycol-6000 (PeG-6000), where the percentage of
germinated seeds was very low and the biomass
was insufficient to perform the determinations.
α-amylase enzymatic activity. The enzymatic
extract was cold performed by homogenization of
the plant material in a buffer solution of sodium ci-
trate pH 5,0 in a proportion 1:2 (w/v). The mixture
was centrifuged during 10 minutes at 10 000 rpm
and 4 ºC. The supernatant was collected for deter-
mining the α-amylase activity. A quantity of 0,1 mL
of the enzymatic extract was added to 0,4 mL, a
solution of 1 % starch (w/v) in sodium phosphate
buffer 20 mmo/L pH 6,9 and it was left to react
during 10 min at 37 ºC. The reaction was stopped
with the addition of 0,5 mL of 3,5-dinitrosalicylic
acid. Afterwards, the reacting mixture was heated
at 100 ºC during 10 minutes and 1,2 mL of distilled
water were added to it. The absorbance was deter-
mined at 546 nm, and the enzymatic activity was
expressed as µmoles/min of glucose released per µg
of protein at pH 6,9 and 37 ºC.
The enzymatic activity (eA) was calculated
through the following formula:
Where:
Rde: Rate of daily emergence, calculated as the
percentage of the accumulated emergence divided
by the number of days since the beginning of the
test.
N: Frequency or number of Rde which was cal-
culated during the test
ef: Percentage of seedling emergence at the
end of the 8 days of the test.
Morphological indicators. The length of the
root and the aerial part of the seedlings eight days
after the beginning of the germination experiment
was determined, with the use of paper graduated in
millimeters and the values were expressed in cen-
timeters.
Where:
t: Time of the essay
TV: Total volume of the essay (9,5 mL)
Ve: Volume of the sample (0,1 mL)
df: Dilution factor of the enzymatic extract
All the described spectrophotometric mea-
surements were made in a UV/VIS Ultrospec 2000
spectrophotometer (Pharmacia Biotech, Sweden).
Content of reducing sugars. The content of re-
ducing sugars was determined by the method of di-
nitrosalicylic acid and D-glucose (Sigma) was used
as pattern sugar (Miller, 1959). The absorbance values
were obtained at a wavelength of 456 nm and the
concentration was expressed in mg/mL from the
pattern curve.

Content of total soluble proteins. The protein
content was determined colorimetrically through
the method described by Lowry et al. (1951), with
the use of bovine serum albumin (BSA) as pattern.
The absorbance values were obtained at 750 nm
and the concentrations (mg/mL) were determined
by the pattern curve.
Content of soluble phenols. One hundred milli-
grams of root and aerial part were homogenized
in 1 mL of methanol and were centrifuged at
12 000 rpm. The supernatant was collected for
the colorimetric determination of soluble phenolic
compounds (Friend, 1992). To determine the con-
centration of these compounds chlorogenic acid
(0,05 mol/L) was used as pattern and the absor-
bance values were determined at 725 nm.
Statistical analysis. The data were processed
with the statistical package SPSS®
version 15.0 for
Windows. The adjustment of the data to a normal
distribution was determined by the Kolmogorov-
Smirnov goodness of fit test and variance
homogeneity through Bartlett’s tests (Sigarroa,
1985). The data were processed through simple
classification Anova and Duncan’s multiple range
test was carried out for the comparison among
means (p ≤ 0,05).
Results and Discussion
Germination percentage. The high concentra-
tions of polyethylene glycol in the medium caused
a decrease in the germination percentage of S. bi-
color (fig. 1). The highest values in this indicator
were obtained with 3 % of the polymer, followed
by 9, 6 and 0 %. The high contents of PeG-6000
in the medium (18 and 21 %) remarkably decreased
the germination percentage. The negative effect of
polyethylene glycol on this indicator is related to
a reduction of the solute potential of the medium
and, consequently, of the hydric potential. The de-
crease of the hydric potential affects the imbibition
process and water availability for the seeds, which
is fundamental for the hydration of enzymes and
substrates that participate in the diverse biochemi-
cal reactions which, in turn, trigger the germination
process (Swapna and Rajendrudu, 2015; Fathi and
Tari, 2016).
The decrease in germination indicators such
as germination percentage because of the polye-
thylene glycol, has been reported in different species,
such as: S. bicolor (Rezende et al., 2017), Triticum
durum Desf. (Khayatnezhad and Gholamin, 2011),
Pennisetum glaucum L. (Sani and Boureima, 2014)
andPongamiapinnata(L.)(Swapna and Rajendrudu,

2015). In these works the responses of the seedlings
depended on the genotype (variety) in question.
The results also coincide with the ones observed
in alfalfa (Medicago sativa L.) seedlings, where
germination, growth and tolerance to osmotic stress
induced by polyethylene glycol-6000 in different
concentrations and in three different genotypes:
“Dk166”, “Verdor”, susceptible to water deficit,
and “Salina”, resistant, were evaluated (Castroluna
et al., 2014). The results showed a delay in the
germination process in all the varieties, although
with differences among the evaluated genotypes.
In this research, the fact that cultivar S. bicolor cv.
UDG-110 germinated in a solution of 18 and 21 %
PeG-6000 could suggest the presence of drought
tolerance mechanisms.
Effect of polyethylene glycol on the germination
value of the seedlings. Polyethylene glycol also
affected the vigor of S. bicolor seedlings (fig. 2). The
3 % PEG-6000 solution significantly increased this
indicator with regards to the control; nevertheless,
the treatments with high PeG-6000 concentrations
significantly affected the germination value. The
increase of this indicator in the treatment with
3 % PeG could be related to a higher emergence
rate, associated with higher metabolic activity
in the root, which is the organ directly in contact
with the stressing agent. The increase in the
content of polypeptides or other osmotically active
compounds can compensate the difference of hydric
potential that is generated between the medium
and the radicle. On the other hand, low PeG-
6000 concentrations could induce the expression
of anti-stress genes, as well as many enzymes
that accelerate the metabolic reactions during the
germination stage.
The osmotic stress caused by polyethylene
glycol-4000 (–0,2 MPa) during the germination
process of Vigna unguiculata (L.) Walp., also
decreased several germination indicators, such as
germination energy, daily mean germination and
vigor index (Jain and Saxena, 2016). Kulkarni et al.
(2014) referred a decrease of different germination
indicators of Coriandrum sativum L. under hydric
stress conditions with PeG-6000. In this work
the vigor index progressively decreased with the
increase of the polyethylene glycol concentration:
control (883,04), 5 % (396,67), 10 % (92,13) and 15 %
(0,00).
Effect of polyethylene glycol on root and aerial
part length. The effect of PeG-6000 on sorghum
seedling growth is shown in figure 3. The roots as
well as the aerial part significantly decreased their
length.Inthecaseoftherootstherewerenodifferen-
ces between the control and 3 %, but the higher
concentrations caused an inhibition of growth and
the lowest values were observed in 18 and 21 % of
PeG-6000.
Similarly, the aerial part length did not show
changes between the control and 3 % PeG-6000.
The concentrations between 6 and 15 % of the os-
motic agent affected the length of this organ, while
the higher percentages caused total inhibition of its
growth. These results proved that the aerial part was

more sensitive to the hydric stress than the roots in
this cultivar, which is in agreement with the results
obtained by Gholami et al. (2010) in Prunus spp.
seeds during the germination process with different
PeG concentrations.
Kulkarni et al. (2014) observed a similar effect
of PeG-6000 (0-20 %) on the root and aerial part
length of Coriandrum sativum L. seedlings. With
15 % (–30 MPa) of polyethylene glycol root and ae-
rial part growth completely stopped.
Growth inhibition of the seedlings is directly
related to the presence of PeG-6000 in the medi-
um, which reduces the input of water to the tissues,
which is essential to develop turgor pressure and
for cell lengthening to occur. The inhibition of this
second process under severe hydric stress condi-
tions was observed in different genotypes of Oryza
sativa L. by other authors (Pirdashti et al., 2003).
On the other hand, if the hydric status of the
embryo is affected the metabolism in general
decreases, due to a reduction in the activity of
the enzymes that participate in different vital
processes, such as degradation of food reserves and
cell respiration. The latter is important, because
the fast growth during germination and later stages
demands high energy intake to perform all the
synthesis processes.
Α-amylase activity. The effect of different con-
centrations of PEG-6000 on the α-amylase activity
of S. bicolor seedlings eight days after germination
is shown in figure 4. As can be observed the enzy-
matic activity in the treatments with PeG-6000 was
higher than the control. The highest values were
obtained in the treatment with 9 % of the osmotic
agent, followed by the medium with 6 % PeG-
6000, which was higher than the variants of 3 and
12 %. The lowest values were obtained in the high-
est PeG-6000 concentrations (15 and 18 %). With
21 % PeG-6000 no results were referred, due to the
low germination and seedling growth percentage,
which reduced remarkably the quantity of biological
material to conduct the biochemical determinations.
Similar results were observed by Li et al. (2017)
in Zea mays L. cv. zhengtian 68, who determined
higher α-amylase activity in seeds germinated in the
presence of PeG-6000 compared with the control.
On the contrary, in works conducted by Muscolo
et al. (2014) with varieties of Lens culinaris L., a
decrease in the α-amylase and α-glycosidase, which
participate directly in the germination process,
was shown. The inhibition of these enzymatic
activities was more remarkable in the sensitive
genotypes Ustica and Pantelleria. The decrease
in the enzymatic activity in the presence of high
PeG-6000 concentrations can be related to the
negativeeffectofthepolymerontheenzymestructure,
because polyethylene glycol is a highly hydrophilic
compound that affects the hydration shell of proteins
and protein precipitates are formed (Sim et al., 2012).
The increase of the amylolytic activity in the
presence of polyethylene glycol could be related
to mechanisms of hydric stress tolerance. The in-
crease in the α-amylase activity was used as a cri-
terion to characterize the response of the genotypes
to environmental stresses (Jamil et al., 2006; Oth-
man et al., 2006).

Reducing sugars and total soluble proteins.
The content of reducing sugars showed variations
in the treatments with different polyethylene glycol
levels (fig. 5). In the roots of the seedlings treated
with 3 and 6 % PeG-6000 a higher concentration
was observed compared with the control. In the
treatments with 9, 12 and 15 % no differences were
observed with regards to the control, while the low-
est values were obtained with 18 % of the product.
The concentration of reducing sugars in the
aerial part showed higher values in the treatments
with 6, 9 and 12 % PeG, and the treatment with
9 % was significantly higher than 6 % of the osmotic
agent. The values in these variants were higher than

the ones observed in the treatments with 3 and 15
%, and the control (without differences among
them), while the lowest values were obtained with
18 % PeG-6000.
These results could be related to the increase
of the α-amylase activity in the presence of PEG-
6000, as was observed in some treatments such as
in 3 and 6 % (aerial part), and 6, 9 and 12 % in the
roots. On the other hand, the increase in the lev-
els of reducing sugars can also be associated with
the increase in the expression of other hydrolytic
enzymes under osmotic stress conditions (Thal-
mann and Santelia, 2017). The sugars released by
the catalyst action of these enzymes may constitute
an osmoprotective mechanism to increase osmotic
pressure in plant tissues and prevent water loss, as a
consequence of the decrease of the hydric potential
in the medium imposed by polyethylene glycol. The
decrease in the reducing sugar contents of the aerial
part, in the treatment with 18 % PeG-6000, can be
related to an increase of maintenance respiration in
these seedlings; that is, in order to maintain a lev-
el of metabolic activity, sugar intake increases for
obtaining energy used in cell refill and synthesis of
proteins, enzymes and other compounds that par-
ticipate in anti-stress response.
The results of this research are in correspon-
dence with the report by Neto et al. (2009). These
authors found an accumulation of soluble carbohy-
drates, sucrose, glucose and fructose in the tissues
of Sorghum sp. leaves during vegetative develop-
ment. This suggests that plants are capable of adapt-
ing to osmotic changes, due to a fast degradation of
starch and formation of soluble sugars during the
vegetative stage.
Figure 6 shows the content of total soluble
proteins in the seedlings germinated under hydric
stress conditions. In the roots the highest values
were observed in 3 and 6 % PeG-6000, without
differences between both; however, the treatment
with 3 % was higher than the control and the
variants with 9, 12 and 15 % (without differences
among them). The lowest content was obtained with
18 % polyethylene glycol in the medium. Regarding
the aerial part, the highest contents were obtained
with the variant of 6 % PeG-6000, followed by 9
and 12 %, among which there were no differences.
The other treatments showed soluble protein
contents lower and similar among them.
The increase in the content of soluble proteins
in the aerial part in the presence of 6, 9 and 12 %
PeG-6000, can be associated with an increase in the
synthesis of specific proteins that participate in the
anti-stress response (Lum et al., 2014; Shayanfar et
al., 2015; Chen et al., 2018), which could indicate
hydric stress tolerance mechanisms, because the
seedling is capable of increasing the protein metabo-
lism depending on several possible functions such
as cell refill and growth. Nevertheless, in similar
trials with P. pinnata seeds subject to hydric stress
with PeG-6000, a decrease was observed of the
content of total soluble proteins with regards to the

control; although the concentrations of sugars and
free proline significantly increased. This last result
was associated to defense mechanisms of seedlings
to counteract the effect of the osmotic shock exert-
ed by the medium on the plant material (Swapna
and Rajendrudu, 2015).
Soluble phenols. The concentration of soluble
phenols in the aerial part of the seedlings showed
the highest values in the treatment with 18 % PeG-
6000, followed by 15 %, which was higher than 12
%. Between this last variant and the concentrations
of 9, 6 and 3 % there were no differences; however,
the content of polyphenols was higher in 12 % with
regards to the control (fig. 7). Regarding the root,
the highest values were obtained in 3, 6, 9 and
12 % of polyethylene glycol, among which there
were no differences. The polyphenol contents in
the treatments of 6 and 9 % were in turn higher
than the ones observed in the variants of 15 % and
the control (without differences), while the lowest
values were obtained with 18 of the polymer.
The increase of polyphenols in the plants in the
presence of abiotic stresses, such as the hydric and
saline ones, was referred as an antioxidant defense
mechanism (Apel and Hirt, 2004). This could be re-
lated to the overexpression of transcription factors,
which induce the synthesis of enzymes that par-
ticipate in the production of phenolic compounds.
Among them is phenylalanine ammonia-lyase,
which constitutes the main enzyme in the metabolic
pathway of polyphenolic compounds (Baâtour et
al., 2013).
These substances have the capacity to eliminate
the oxygen reactive species (ROS), which affect
drasticallycellmetabolismandprotectthebiological
membranes from the oxidative damage caused by
these radicals (Chernane et al., 2015). On the other
hand, phenolic compounds can form complexes
with the metals that catalyze oxygenation reactions
and inhibit the activity of oxidative enzymes.
In addition, the antioxidant efficiency is much
higher than that of the compounds α-tocopherol
and ascorbate, also known for their antioxidant
properties (Blokhina et al., 2003).
Conclusions
Polyethylene glycol-6000 affected the germi-
nation process of S. bicolor cv. UDG-110, although
the seeds germinated in high concentrations (21 %),
proving the presence of drought tolerance mecha-
nisms.
The presence of PeG-6000 in the medium in-
creased the levels of reducing sugars in the seed-
lings, probably due to the increase of the α-amylase
activity in the tissues and other enzymes related to
carbohydrate metabolism.

The increase in the content of total soluble pro-
teins in the aerial parts of the seedlings germinated
in intermediate PEG-6000 concentrations and the
increase in the concentration of polyphenolic con-
centrations in the treatments with severe hydric
stress, suggest the presence of biochemical mecha-
nisms of anti-stress responses to attenuate the damage
by osmotic and oxidative stress, respectively.
Acknowledgements
The elaboration and execution of this work
took place with the funding provided by the project
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Received: May 16, 2018
Accepted: November 8, 2018

236 Pastos y Forrajes, Vol. 41, No. 4, October-December, 236-240, 2018 / Yuseika Olivera-Castro
Scientific Paper
Population performance of a collection of Urochloa brizantha (Hochst.
ex A. Rich.) R.D. Webster in association with Stylosanthes guianensis
CIAT-184 (Aubl.) Sw
Yuseika Olivera-Castro1
, Pedro Pablo del Pozo-Rodríguez2
, Lisset Castañeda-Pimienta1
and Juan
Francisco Ramírez-Pedroso3
1
2
Universidad Agraria de La Habana Fructuoso Rodríguez, Mayabeque, Cuba
3
Estación Experimental de Pastos y Forrajes Cascajal, Villa Clara
E-mail:yuseika@ihatuey.cu
Abstract
The objective of this study was to evaluate the structural and floristic composition performance of 19 accessions
of Urochloa brizantha, associated with Stylosanthes guianensis CIAT-184 on an acid soil, after three years of simulated
grazing. The variables live shoots-dead shoots ratio and floristic composition were analyzed. The measurements were
made in all the rotations during the experimental period. The substitution of dead shoots was favorable in most of the
treatments; four accessions stood out (CIAT-16322, CIAT-26646, CIAT-16332 and CIAT-16335) in which, at the end of
the period, the index of substitution of dead shoots was over two. From the accessions, 36,6 % reached a percentage higher
than 80 % at the end of the experimental period; the accessions CIAT-16317, CIAT-16335 and CIAT-16332 were the ones
with higher presence of the base pasture (> 90 %). In all the associations the legume was represented in a low percentage
(< 16 %); while the weeds remained, in most cases, below 20 %. In general, the evaluated U. brizantha accessions showed
an acceptable persistence or stability in time, and CIAT-16322, CIAT-26646, CIAT-16332 and CIAT-16335 stood out.
Keywords: botanical composition, grasses, legumes, shoot
Introduction
Although the agronomic and nutritional value
variables are important in the process of plant
selection, it is considered essential to complement
thisinformationwiththatfromnolesstranscendental
variables, such as floristic composition and live
shoots-dead shoots ratio (Ls/Ds); the latter as
component of the pasture structure (Olivera,
2016). In addition, an analysis of the variation of
the structural indicators of pasture and of floristic
composition provides elements to discuss about the
possible contrast among the treatments for a certain
condition.
Thus, it is necessary to know the performance
of pasture, because the inadequate use of the pas-
tureland –in terms of under- or overgrazing or
due to ignorance– affects cattle production (Vane-
gas-Moreno, 2015); because pasture, as feeding
basis, contributes to cattle the necessary feedstuff
and energy (Pintado-Lazo and Vásquez-Rodríguez,
2016).
The association of pasture with legumes is
beneficial, because they play an important role
in the fixation and subsequent increase of N in
the associated forage; this has been the object of
research, which has allowed to conclude that the N
fixed by legumes shows values between 50 and 300
kg N/ha/year (Carrero, 2012). For such reason, the
association of legumes with grasses is an excellent
alternative to fix this element and make it available
for the associated species, with which higher forage
production is achieved (Luengas-Barrera and Hena-
Ruiz, 2016).
In addition, it is known that the systems with
legumes –herbaceous or shrubby–, associated with
grasses, can play a very outstanding role in the eco-
system, due to it and other qualities of those species.
In that sense, Leucaena leucocephala (Lam). de
Wit, Teramnus labialis (L.f.) Spreng., Centrosema
molle Mart. ex Benth., Stylosanthes guianensis
(Aubl.) Sw., among others, stand out.
For such reason, the objective of this study was
to evaluate the structural and floristic composition
performance of 19 accessions of Urochloa brizantha,
associated with S. guianensis CIAT-184 on an acid
soil, after three years of simulated grazing.

PastosyForrajes,Vol.41,No.4,October-December,236-240,2018 / Population performance of Urochloa brizantha withStylosanthesguianensis 237
The study was conducted in areas of the
Cascajal Pastures and Forages Research Station,
located in the Santo Domingo municipality –Villa
Clara province, Cuba–, at 22º 36’ North latitude
and 80º 04’ West longitude, at 60 m.a.s.l.; on a
petroferric dystic Gley Nodular Ferruginous, which
shows acid pH (4,2), as well as low organic matter
contents (1,91 %), total N (0,40 %) and assimilable P
(1,90 mg/100 g). According to these characteristics,
it is considered as an acid and low fertility soil
(Hernández-Jiménez et al., 2015). The climate
variables are shown in table 1.
Experimental procedure
Design and treatments. A randomized block
design with three replicas was used. The plots
measured 23,52 m2
and were separated by 1,50-m
spaces in both senses.
The treatments were represented by the 19
U. brizantha accessions, previously selected from a
study carried out before: CIAT-16300, CIAT-16317,
CIAT-16809, CIAT-16469, CIAT-16322, CIAT-
16132, CIAT-16128, CIAT-16335, 1539, CIAT-
26290, CIAT-16332, CIAT-16819, CIAT-16303,
CIAT-16334, CIAT-16448, CIAT-26646, CIAT-
16485, CIAT-16197 and CIAT-26032.
Measurements. The following measurements
were made at the end of the experimental period,
which was of three years of evaluation under
simulated grazing, in which 15 rotations were
carried out. For such purpose the recommendations
made by Machado et al. (1999) were followed:
• Live shoots-dead shoots (Ls/Ds) ratio. It was de-
termined by the physical count of the number of
live shoots and dead shoots in two tillers from
each plot, in the three replicas, for all the acces-
sions. To count the dead shoots those which were
in a status of total deterioration, that is, necrotic
or detached from the tiller, were considered. In
this case, the sample size was 4,25 %.
• Floristic composition. To determine the floristic
composition of the existing species in the flora
of each one of the associations, a 1,0-m2
frame
was used divided into four quadrants. In each
of them the percentage of weeds was estimated,
as well as the percentage of U. brizantha and S.
guianensis CIAT-184. Two samplings were sys-
tematically performed per plot in each replica,
which represented a sample size of 8,5 % of the
whole plot.
Statistical analysis. For the data processing
variance analysis was carried out from a model
that included the effects of the accessions and/or
associations and the replicas. Before it the normal
distribution was tested through the Kolmogorov-
Smirnov test, and the variance homogeneity, by
Levene’s test. The means were compared through
Duncan’s multiple range test, for a significance level
of p < 0,05. For such purpose the statistical package
SPSS®
(version 15.0)was used.
Table 2 shows the results of the studied variables
(Ls/Ds ratio and floristic composition of the pas-
tureland) in each of the studied associations. Thir-
teen of the 19 evaluated accessions (68,4 %) ended
with a favorable Ls/Ds ratio, that is, with a produc-
tion rate of live shoots over 1 (Machado, 2002);
CIAT-16335, CIAT-16332 and CIAT-26646 stood
out for ending with a value higher than 2, show-
ing highly significant differences (p < 0,001) when
compared with the other treatments. Nevertheless,
although CIAT-16322 also finished the experimen-
tal period with an index higher than two, it showed
statistical differences with regards to the three
above-mentioned ones.
Such performance indicates that these acces-
sions were capable of maintaining a favorable pro-
duction rate of live shoots, particularly the most
outstanding ones, and, thus, keeping an acceptable
density of the new material. This favors the substi-
Table 1. Performance of climate during the research.
Year
Rainfall
(mm)
Variable
Temperature (ºC) Relative humidity
(%)
Hours light
(h)Maximum Minimum Mean
1 1 697,7 31,5 19,5 24,4 78 7,9
2 1 153,5 31,3 19,2 24,3 76 8,0
3 1 262,3 30,2 18,5 23,6 77 7,8

tution of dead shoots by live shoots, and this way
the maintenance of the integrity of tillers is en-
sured, which may be considered as an adaptation
element of such accessions.
In this sense, Ramírez-Reynoso et al. (2011)
claimed that the production and survival of
shoots is a mechanism used by plants to maintain
persistence, which depends on their capacity
to remove dead shoots and keep the population
density of stems stable, aspect that contributes
decisively in the stability of the pastureland and
which is directly determined by the combined
effect of seasonal patterns of the emergence, death
and survival processes of those plant components
(Ramírez-Reynoso et al. 2011).
These authors also referred to the fact that there
are remarkable differences among species and culti-
vars to achieve stability of the population density of
the shoots, and, thus, favor pastureland persistence.
For such reason, they stated that the variations that
occur in the emergence, death and survival rate of
the shoots are valuable to understand the mecha-
nisms that are involved in pasture persistence and
regrowth, which is considered important in the
selective process. In addition, these variations in
plant structure may be influenced by genetics, plant
physiology and interaction with the surrounding
environment (climate variables and soil).
However, although the rainfall volume and
other climate variables, such as temperature, rela-
tive humidity and hours light, were propitious for
the production of new shoots, it was observed that
some of the accessions showed a very low produc-
tion index of new shoots. Among them were 1539,
CIAT-16809 and CIAT-16469, with 0,42; 0,52 and
0,54, respectively; which did not differ statistically
among them, but they did differ from the other ac-
cessions, and showed a marked trend towards tiller
degradation and, subsequently, pastureland degra-
dation.
The existence of other accessions with better
replacement index than the above-mentioned one
(lower than 0,55), but which, similarly, reached
values below one, was also noted, as in the case of
Table 2. Performance of the Ls/Ds ratio and floristic composition of the pastureland.
Accession Ls/Ds Grass S. guianensis Arvenses
CIAT-16300 1,24cd
80,00h
16,30h
3,70abc
CIAT-16322 2,09f
84,00i
10,03cdefgh
6,00bc
CIAT-16819 1,85e
66,07fg
4,80abcde
20,07d
CIAT-16334 1,21cd
60,57cd
6,37abcdef
26,87e
CIAT-26646 2,27g
83,83i
12,20fgh
4,03abc
CIAT-16197 0,78b
64,20ef
2,47ab
20,60d
CIAT-16809 0,52a
51,47b
9,23cdefg
30,33e
CIAT-16128 1,30d
60,00cd
10,03cdefgh
15,97d
CIAT-16332 2,29g
96,87l
1,70a
1,43ab
CIAT-16317 1,10c
91,93k
4,27abcd
3,77abc
CIAT-16132 1,08c
45,47a
11,13efgh
35,73f
CIAT-26290 0,74b
58,37c
10,20defgh
28,40e
CIAT-16303 0,82b
62,53de
9,40cdefg
26,13e
CIAT-16448 1,23cd
59,17cd
8,13abcdef
26,17e
CIAT-16485 1,85e
78,03h
15,00gh
7,00c
CIAT-26032 1,36d
68,93g
6,00abcdef
16,97d
CIAT-16469 0,54a
88,50j
4,40abcde
6,13bc
CIAT-16335 2,30g
96,47l
3,33abc
0,20a
1539 0,42a
62,53de
8,67abcdefg
28,80e
SE ± 0,082*** 1,11*** 1,99*** 1,64***
a, b, c, d, e, f, h, g: means with different letters in the same column significantly differ at
p < 0,05, (*** p < 0,001).

PastosyForrajes,Vol.41,No.4,October-December,236-240,2018 / Population performance of Urochloa brizantha withStylosanthesguianensis 239
CIAT-16303, CIAT-16197 and CIAT-26290, which
did not differ among them. This is considered as an
element that limits the possible selection of all those
materials as a whole.
When analyzing the values in floristic com-
position of the pastureland, it was proven that the
associations made up by accessions CIAT-16332
and CIAT-16335 concluded the experimental pe-
riod with more than 95 % of the area covered by
the grass, with which they were significantly higher
(p < 0,001) than the other evaluated ones.
These accessions were followed, in descending
hierarchical order, by CIAT-16317, CIAT-16469,
CIAT-16322, CIAT-26646, CIAT-16300 and CIAT-
16485, among which highly significant differences
(p < 0,001) were also found, but all of them with a
percentage of covered area that varied between 78,0
and 91,9 %.
The results showed that all those accessions
concluded the experimental period with high values
of area covered by the grass, which indicates that
they kept outstanding population stability after 15
rotations under those conditions. These values are
considered an adequate and, in turn, recommended
index for the selection and proposal of the possible
commercial varieties (Machado et al., 1999).
In the accession CIAT-16132 the lowest covered
area percentage (45,4 %) was detected, although
other two (CIAT-16809 and CIAT-26290), also
with highly significant differences between them
(p < 0,001), showed values of 51,4 and 58,3 %; and,
in turn, values below one in terms of the Ls/Ds
ratio; which contrasts with the performance main-
tained by the most advantageous materials, as was
previously discussed.
In turn, in the associations in which the grass
had a lower percentage in the floristic composition
the weeds increased (table 2). It is known that these
species develop mechanisms that facilitate the
colonization of empty spaces that the pasture does
not occupy during the establishment or exploitation
period (Sardiñas et al., 2015).
The association in which the accession CIAT-
16335 was found, without differing from CIAT-
16332, CIAT-16317 and CIAT-26646, showed the
lowest area invaded by the other weed species
(0,20 %); while the association made up by the
accession CIAT-16132 was the most invaded one,
although the absolute value did not exceed 35,73 %.
According to Padilla et al. (2013), when the invasion
surpasses 65 %, the floristic composition of the
pastureland and the pasture yield are considerably
affected. In this case, in none of the associations the
weeds exceeded 60 % (table 2).
On the other hand, S. guianensis CIAT-184
is one of the legumes which have shown better
adaptability attributes under the acidity conditions
of Cuban soils, and is among the varieties reported
by MINAG (2017) as commercial, for which it was
chosen to establish the association in this research.
According to the results (table 2), highly
significant results (p < 0,001) were found in the
area occupied by S. guianensis. The highest value
was detected in the association with CIAT-16300
(16,30 %); it did not differ from the one found in the
associations made up by CIAT-16485, CIAT-26646,
CIAT-16132, CIAT-26290, CIAT-16322 and CIAT-
16128, in which the percentage of this legume
varied in the range from 10,0 to 15,0 %.
The lowest values were found in the associa-
tions with CIAT-16332, CIAT-16197, CIAT-16335,
CIAT-16469, CIAT-16819 and CIAT-16317, all with
less than 5 % and without significant differences
among them.
Thus,itispossibletostatethattherepresentation
of S. guianensis in the floristic composition, in all
cases, was little consistent, particularly in the last
rotations in which values below 10 % were observed
(table 2); because for the legumes in the association
to have impact, they should be in a proportion never
lower than 30 % (Roca-Cedeño et al., 2014).
Some of the factors that contributed to the
detriment of the population of S. guianensis could
be related with the growth habits of the grass,
which is tillering and erect; for which it is not easily
associated with non-twining herbaceous legumes
(Smith, 2014).
In addition, Traveset (2015) asserted that the
character of the competition and competitive ability
of one species not only depend on the species and
its needs, but also on the environmental conditions,
and change with them.
It is also known that, from the physiological
point of view, grasses show a series of advantages
with regards to legumes, among which the higher
photosynthesisratecanbementioned,whichconfers
them higher growth and development (Vanegas-
Moreno, 2015; Gutiérrez-Guiñan, 2016); as well as
higher tolerance to high temperatures (Pozo et al.,
2011), allowing them to compete favorably.
S. guianensis, even when it was maintained
throughout the exploitation period, did it with low
percentage and showed a strong trend to disappear;
especially if it is taken into consideration that its
population at the beginning of the experimental
stage fluctuated between 31,2 and 36,3 %, depending

on the criterion for considering the plot area as es-
tablished (Machado et al., 1999).
It is concluded that, for the analyzed population
variables, the associations made up by the acces-
sions CIAT-16322, CIAT-26646, CIAT-16332 and
CIAT-16335 stood out, which have potential for
their utilization on acid soils.
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Received: August 3, 2018
Accepted: October 22, 2018

Pastos y Forrajes, Vol. 41, No. 4, October-December, 241-247, 2018 / Edaphic macrofauna in animal husbandry systems 241
Scientific Paper
Performance of the edaphic macrofauna in animal husbandry systems,
in a productive entity of the Yaguajay municipality, Cuba
Wendy Mercedes Ramírez-Suárez1
, Marta Beatriz Hernández-Chávez1
, Alexis Abilio Zurita-
Rodríguez2
and Marlen Navarro-Boulandier1
1
Estación Experimental de Pastos y Forrajes Indio Hatuey, Universidad de Matanzas. Ministerio de Educación Superior
2
Estación de Protección de Plantas, Iguará, Yaguajay. Sancti Spíritus
E-mail: wendy.ramirez@ihatuey.cu
Abstract
A study was conducted in areas of a basic unit of cooperative production, of the Yaguajay municipality –Sancti
Spiritus province, Cuba–, in order to evaluate the edaphic macrofauna in two animal husbandry systems: natural
pastureland and silvopastoral system, on a vertic Gleysol soil. The soil was sampled during two years in the rainy and dry
seasons, in the litter and at the depths 0-10, 10-20 and 20-30 cm, according to the methodology of the Tropical Soil Biology
and Fertility International Research Program. The macrofauna was identified to the taxonomic category of order. In both
seasons and systems three phyla, seven classes and 11 orders were found. A total of 1 207 individuals were collected;
from them, 840 corresponded to the silvopastoral system and 367 to the pastureland. In both systems coleopterans
prevailed (36 and 37 % for the pastureland and the silvopastoral system, respectively), followed by Oligochaeta (21 and
17 %, respectively). A higher density of individuals was obtained in the silvopastoral system. It is concluded that the
taxonomic composition of the edaphic macrofauna was similar in the evaluated systems, with differences regarding the
diversity of the orders. The highest quantity and density of individuals were obtained in the silvopastoral system. The
effect of depth on the macrofauna composition and the distribution of the collected individuals, as well as the preference
for the shallowest layer of the soil, was proven.
Keywords: biota, natural pastureland, silvopastoral system
Introduction
The deterioration of soil quality is one of the
most serious problems that animal husbandry sys-
tems have experienced in recent years, mainly due
to inadequate agricultural practices.
The most significant problems in soil conser-
vation at global scale, according to FAO (2016), are
erosion, loss of organic carbon and nutrient unbalance.
Animal husbandry utilizes those soils of lower pro-
ductive value and, thus, in them the above-men-
tioned characteristics are increased (Lok, 2016).
Romanuk et al. (2014) sustain that the soil consti-
tutes a unique biological system where there is a
large diversity of organisms which play multiple
key functions for the ecosystems. In this sense,
Veresoglu et al. (2015) stated that soil biodiversity
should also be considered as keeper of food security
and of the ecosystem services in the face of climate
change, because it propitiates a more complex and
much more resistant structure.
The edaphic biota plays an important role in
the biogeochemical processes of the soil in animal
husbandry systems, and its functions include: litter
decomposition, nutrient recycling, synthesis and
mineralization of organic matter and modification
of the soil structure, among others; this influences
the system integrity and productivity (Sánchez et
al., 2011).
In animal husbandry ecosystems the knowledge
of the edaphic biota has particular interest in
Cuba, due to the functions it plays in the systems
that are used for cattle feeding. In the case of the
Yaguajay municipality, and specifically in the basic
unit of cooperative production (UBPC) La Elvira,
no reference was found about the macrofauna
behavior, very important indicator for evaluating
the soil quality. For such reason, the objective of
this study was to evaluate the edaphic macrofauna
composition and behavior in two systems: natural
pastureland and silvopastoral system, in a vertic
Gleysol soil.
Characterization of the study area. The
research was conducted in areas of the UBPC La
Elvira, belonging to the agricultural enterprise

242 Pastos y Forrajes, Vol. 41, No. 4, October-December, 241-247, 2018 / Wendy Mercedes Ramírez-Suárez
Obdulio Morales, in the Yaguajay municipality –
Sancti Spiritus province, Cuba–. This productive
entity has a total area of 1 878 ha and from them
720 ha are aimed at animal husbandry.
Edaphoclimatic conditions. The soil where the
experimental stage was conducted belongs to the
grouping of Gleysols and to the genetic type vertic
Gleysol, according to the classification proposed by
Hernández-Jiménez et al. (2015), and shows clayey
texture, effective depth of 0,90 m and deficient sur-
face and internal drainage. Its apparent density is
1,26 g/cm3
, it shows a field capacity of 42,1 % and
its topography is flat.
During the experimental period rainfall varied
between 1 200 and 1 400 mm as annual average,
with two well-differentiated seasons: a rainy sea-
son (May-October) in which 76 % of the rainfall
occurred and a dry season (November-April). The
mean annual temperature was 25,6 ºC.
Treatments. The treatments consisted in two
systems, both with more than 10 years of exploita-
tion:
• Silvopastoral system. In the tree composition the
species Leucaena leucocephala (Lam.) de Wit,
Albizia niopoides (Spruce ex Benth.) Burkart
and Talipariti elatum (Swartz) Fryxell (blue
mahoe) were present; and in the herbaceum
stratum, natural pastures. The global stocking
rate was 1,1 LAU/ha.
• Pastureland system. A pastureland system was
evaluated mainly composed by the species Bo-
thriochloa pertusa (L.) A. Camus and Urochloa
ruzziziensis (R. Germ et Evrard). The area cove-
red by the pastures in all the paddocks exceeded
90 %.
Experimental design. A completely randomized
experimental design was used, with three replicas
per treatment.
Experimental procedure. The macrofauna sam-
pling was performed between 7:00 a.m. and 9:00
a.m. in the two systems, at the end of both seasons
during two years, according to the methodology of
the Tropical Soil Biology and Fertility International
Research Program (Anderson and Ingram, 1993);
it consists in the extraction of monoliths of 25 x
25 x 30 cm in a transept, whose point of origin is
randomly and linearly determined. The following
strata were evaluated: litter, 0-10 cm, 10-20 cm and
20-30 cm. The macrofauna was manually collected
in situ. The earthworms were preserved in 4 % for-
maldehyde, and the other invertebrates, in 70 % al-
cohol, for their later identification in the laboratory.
The macrofauna was identified to the taxo-
nomic category of order, according to the criterion
expressed by Ruiz et al. (2008). The average den-
sity values (ind. m-2
) for the edaphic community,
for each taxon and per stratum were determined,
in each study system. The density was determined
depending on the number of individuals.
Statistical analysis. For the analysis of the
studied variable the fulfillment of the variance ho-
mogeneity (test of Levene, 1960) and normal dis-
tribution (Shapiro and Wilk, 1965) assumptions
was tested; as the homogeneity requisite was not
fulfilled, non-parametric analysis was performed.
To determine the variations of the density of the
edaphic macrofauna between the systems and stra-
ta, the Kruskal-Wallis test was used. The statistical
processing was done with the software InfoStat,
free version for Windows®.
Taxonomic composition of the soil macrofauna
The taxonomic composition of the soil macro-
fauna in the natural pasture system (NP) and in the
silvopastoral system (SPS), for the rainy and the dry
season, is shown in table 1.
In both seasons and systems three Phylum, seven
classes and 11 orders were found; in general, in the
silvopastoral system there was a higher presence of
orders, which could be related to a higher soil cover
in this system, propitiating better temperature and
humidity conditions for the optimum development
of the macrofauna (Cabrera-Dávila et al., 2017).
Such results are higher than the ones found by
Chávez-Suárez et al. (2016), who studied the macro-
fauna in mountain animal husbandry ecosystems in
Guisa, Cuba, and only seven orders were reported
in pasturelands; this could have been related with
the drainage characteristics of the vertic Gleysol
soil, present in the studied areas.
On the other hand, García et al. (2014) reported
the presence of 14 orders in a silvopastoral system
with L. leucocephala on a Ferralitic Red soil, which
could have been due to the fact that their study was
conducted in systems with cultivated grasses and
on another soil type; these plants propitiate a high-
er biomass quantity and maintain the cover, which
creates adequate conditions for the development of
macrofauna biodiversity.
The fact that in the two seasons the same quan-
tity of Phylum, classes and orders was found could
have been related to the soil characteristics in the

Pastos y Forrajes, Vol. 41, No. 4, October-December, 241-247, 2018 / Edaphic macrofauna in animal husbandry systems 243
experimental areas; which is formed from clayey
sediments, with predominance of smectites among
the clayey minerals, and shows gley characteristics at
less than 50 cm of depth and vertic horizon (Hernán-
dez-Jiménez et al., 2015). That is why throughout
the year in this soil type humidity is maintained,
which influences positively the development of the
edaphic biota. According to Siqueira et al. (2016),
the macrofauna community is affected by the hy-
drological regime in the different land uses.
Theearthwormswerepresentinthetwosystems
and in both seasons, which coincides with the report
by Chávez-Suárez et al. (2016) regarding the fact
that, in animal husbandry systems –especially in the
most humid ecosystems and in pasturelands–, there
is a predominance of these macrofauna individuals.
It is important to acknowledge the functionality of
earthworms for the maintenance of the ecosystem
services of the soil, due to their contribution as
physical engineers, because they create channels
and aerate the edaphic medium; in addition, they are
considered biochemical engineers, for promoting
organic matter decomposition and causing
interactions with fungi and bacteria (Lavelle et
al., 2016). These authors emphasize the services
provided by earthworms in the formation of the
edaphic medium through the mixture of organic
and mineral components, and their bioturbation
activities contribute to homogeneization; while
the critical points of the drilosphere (soil zone
influenced by earthworm action) increase spatial
heterogeneity.
Figures 1 and 2 show the percentage distri-
bution of the total number of individuals for each
taxonomic group in the natural pasture and the
silvopastoral system, respectively. As can be ob-
served, the Coleoptera order was the one with the
highest presence in both systems, because it was
represented by 36 and 37 % of the total, respective-
ly, followed by Haplotaxida (21-17 %) and Isopoda
(14-14 %); similar results were reported by Cabrera
et al. (2011) and García et al. (2014). The other or-
ders did not exceed 5 %, except Orthoptera with 11
% in natural pastures, and Orthoptera and Geophi-
lomorpha with 6 % in the silvopastoral system.
The higher presence of coleopterans in both
systems is important, because due to their wide
variety of feeding habits and biotic preferences
they have ecological and economic repercussion on
agroecosystems. In this sense, Cabrera-Dávila et al.
(2017) stated that, according to their functionality,
theycanbedetritivorous,predatorsandherbivorous;
and they also show high abundance and diversity of
species.
Density of the edaphic macrofauna
Figure 3 shows the average density values in
litter for each season and system. The best perfor-
mance was found in the silvopastoral systems for
both seasons, in which there was higher density,
Table 1. Taxonomic composition of the macrofauna.
Phylum Class Order
Natural pastures Silvopastoral system
Year 1 Year 2 Year 1 Year 2
RS DS RS DS RS DS RS DS
Arthropoda
Insecta
Coleoptera X X X X X X X X
Lepidoptera X X X X X X X
Hemiptera X X X X X X X
Orthoptera X X X X X X X X
Diptera X X X X X X X
Arachnida Araneae X X X X X X X
Chilopoda Geophilomorpha X X X X X X X
Diplopoda Spirobolida X X X X X X
Malacostraca Isopoda X X X X X X X X
Mollusca Gastropoda Archaeogastropoda X X X X X X X
Annelida Clitellata Haplotaxida X X X X X X X X
X: presence, DS: dry season, RS: rainy season.

244 Pastos y Forrajes, Vol. 41, No. 4, October-December, 241-247, 2018 / Wendy Mercedes Ramírez-Suárez
with significant difference from the natural pasture
system.
These results coincide with the ones obtained
by Cabrera-Dávila et al. (2017), who found high-
er abundance in agroforestry systems and forests,
which was ascribed to a higher diversity of resources
offered by these ecosystems, such as: shade, soil
protection, high edaphic humidity and low tem-
peratures, elements that contribute to the soil life
subsistence.
In that sense, the values of this research could
be ascribed to the higher presence of foodstuffs in
the litter layer which, gradually, is formed on the
soil with the fall of leaves from the trees, which

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