Journal of the Saudi Society of Agricultural Sciences 21 (2022) 425–431

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Journal of the Saudi Society of Agricultural Sciences

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Extract yield, dilution methods and antifungal potential of fruits of

Picralima nitida (Stapf.) T. A. Durand & H. Durand
Ghislain Comlan Akabassi a,b,c,⇑, Elie Antoine Padonou b,d, Edouard Jacques Kouadio Yao e, Silue Nakpalo e,
Koffi Kibalou Palanga f, Bidossèssi Eliane Juliette Assogbadjo a,b, Martine Zandjanakou-Tachin g,
Achille Ephrem Assogbadjo b, Noël Guede Zirihi c
a
African Center of Excellence for Climate Change, Biodiversity and Sustainable Agriculture, Unviversity Félix Houphouët Boigny, 22 BP 582 Abidjan 22, Cote d’Ivoire
b
Laboratory of Applied Ecology, Faculty of Agronomic Sciences, University of Abomey Calavi, 05 BP 1752 Cotonou, Benin
c
Laboratoire de Botanique, UFR Biosciences, Unviversity Félix Houphouët Boigny, 22 BP 582 Abidjan 22, Cote d’Ivoire
d
School of Tropical Forestry, National University of Agriculture, Kétou, Benin
e
Laboratoire de Physiologie Végétale, UFR Biosciences, Université Félix Houphouët-Boigny, 22 BP 582 Abidjan 22, Cote d’Ivoire
f
Institut Supérieur des Métiers de l’Agriculture de l’Université de Kara, BP 404, Togo
g
Laboratory of Plant Pathology, National University of Agriculture, 01 BP 55 Porto-Novo, Benin

a r t i c l e i n f o a b s t r a c t

Article history: Today, despite its status as a key sector, agriculture faces many problems due to fungi and the use of syn-
Received 19 April 2021 thetic pesticides. This study evaluated the antifungal potential of Picralima nitida fruits against Fusarium
Revised 2 September 2021 oxysporum. The extracts were obtained by the Total Aqueous Extraction method (TEA) and
Accepted 25 November 2021
Hydroethanolic extraction (EE70%) method. Two dilution types of the extracts in PDA medium were
Available online 1 December 2021
tested: (1) dilution before autoclaving (AvA) and (2) dilution after autoclaving (ApA). The extracts of
two fruit morphotypes from two climatic zones were tested at a concentration of 3 mg.ml
1. The yield
Keywords:
of fruit extracts from Dahomey Gap (DG) was significantly higher (>8%) than that from the Guinea-
Dahomey Gap
Picralima nitida
Congolese region (GC). A significant difference was observed in yield (p < 0.05) between TEA type and
Extraction method the EE70% type with a higher inhibition rate (27%) of EE70% on Fusarium oxysporum. No significant differ-
Dilution method ence was observed between the pathogen inhibition rates in the dilution type (p > 0.05). The inhibition
Fusarium oxysporum rate of the pathogen was 25.65% for the short fruit morphotype and 25.08% for the large fruit morpho-
Yield extract type. This study demonstrated possibility of using extracts of Picralima nitida in agriculture.
Ó 2021 The Authors. Production and hosting by Elsevier B.V. on behalf of King Saud University. This is an
open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

1. Introduction less of their stage of development. For example, The Colletotrichum

In most developing countries, agriculture is hampered by many
difficulties such as the lack of arable land, the acquisition of suit-
able materials and the eternal concern about low yields and con-
servation due to pathogens. Among the most important
pathogens, fungi are those that have a broad spectrum and are
more dangerous. They are able to infecting all plant organs regard-
⇑ Corresponding author at: African Center of Excellence for Climate Change,
Biodiversity and Sustainable Agriculture, Unviversity Félix Houphouët Boigny, 22
BP 582 Abidjan 22, Cote d’Ivoire.
E-mail address: cgakabassi@gmail.com (G.C. Akabassi).
Peer review under responsibility of King Saud University.
Production and hosting by Elsevier
gloeosporioides responsible for crops’ anthracnose is able to infect-
ing leaves, flowers and fruits, thus reducing yield and quality. In
tomatoes (Lycopersicon esculentum M.), Pythium sp. and Fusarium
oxysporum can attack all the organs and especially root system,
inducing a loss of up to 60% of the production (Doumbouya et al.,
2012). F. oxysporum is a pathogen common to several crops in agri-
culture (tomato, banana, groundnut, maize, etc.) and difficult to
control.
A wide range of synthetic and biological fungicides have been
developed and used to control the pathogenic strains. The use of
synthetic fungicides is the most widely used method in intensive
production systems. The overuse of these pesticides can have
harmful consequences on human health (e.g. headaches, respira-
tory problems, skin irritation, coughing, fatigue, diarrhea, dizzi-
ness) and on the environment such as groundwater pollution
(Son et al., 2017). The multiple consequences associated with this
method often do not encourage producers to use it (Yarou et al.,

https://doi.org/10.1016/j.jssas.2021.11.006
1658-077X/Ó 2021 The Authors. Production and hosting by Elsevier B.V. on behalf of King Saud University.
This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
Ghislain Comlan Akabassi, Elie Antoine Padonou, Edouard Jacques Kouadio Yao et al.
2017). To these consequences, is added the problem of resistance
of certain pathogenic strains (Houndété et al., 2010; Agboyi et al.,
2016). For an environmentally friendly alternative solution, phyto-
fungicides has been adopted. The method consists of applied aro-
matic and/or medicinal plant extracts as a biological pesticide.
The advantages of the method is a low-cost and environmentally
friendly solution.
Some aromatic and/or medicinal plants contain substances with
therapeutic and antiparasitic properties (Pamo et al., 2004). Among
these, some have already demonstrated their effectiveness in the
agricultural field; for instance, Ocimum gratissimum and Ocimum
basilicum, Melaleuca quinquenervia and Cymbopogon giganteus.
Others such as Picralima nitida (Stapf.) T. A. Durand & H. Durand,
are very rich in alkaloids with fungicidal, bactericidal, acaricidal,
insecticidal properties (Dibua et al., 2013) and have been widely
used in the pharmacological field. They can also be used as
fungicides.
P. nitida is a medicinal plant of the Apocynaceae family; widely
used in traditional medicine. It is known by several names depend-
ing on the country and the socio-cultural group. In the Guineo-
Congolese region of West Africa, it is more locally called
‘‘Akuamma” and ‘‘Ayokpè” in the Dahomey Gap zone. The common
name in French is ‘‘Obéro à gros fruit” (Akabassi et al., 2020). P.
nitida is a shrub that reaches 5–35 m in height. When ripe, the fruit
is yellow or light orange in colour. It is traditionally used to treat
malaria (Adjanohoun et al., 1996; Akabassi et al., 2017), diabetes
(Teugwa et al., 2013; Akabassi et al., 2017; 2021), fevers (Yakeu,
2012), cough (Dapaah, 2016) and infectious diseases in general
(Akabassi et al., 2020).
In pharmacology, previous studies (Duwiejua et al., 2002) have
shown that P. nitida is rich in alkaloids. The methanol fruit extract
has shown an anti-pyrexia effect (Ezeamuzie et al., 1994). The
methanol and chloroform extracts from the seeds were very effec-
tive against ulcers (Kouitcheu et al., 2008). Hydroethanolic,
metanolic, chloroform and aqueous extracts from different parts
of the plant showed effective inhibitory activity against certain
bacteria such as Escherichia coli, Pseudomonas aeruginosa, Bacillus
subtilis, Staphylococcus aureus and Salmonella kintambo (Nkere
Fig. 1. Geographical location of the study area.
426
Journal of the Saudi Society of Agricultural Sciences 21 (2022) 425–431
and Iroegbu, 2005; Dapaah, 2016). The antifungal activity of two
extracts (ethanolic and aqueous) from leaves tested on Aspergillus
flavus, Candida albicans, Microsporum canis confirmed that P. nitida
has effective antifungal activity (Ubulom et al., 2011). hydoethano-
lic and aqueous extracts of P. nitida seeds have strong larvicidal
activity against larvae of Anopheles gambiae (Dibua et al., 2013).
Despite the efficacy of the antimicrobial activity of the species,
no studies have yet been carried out to test the antifungal potential
of P. nitida extracts on mycopathogens. Therefore, more informa-
tion is needed to avoid contamination leading to the loss of prod-
ucts and materials during the test. In addition, as most plant
extracts are contaminated either during the extraction procedure
or the extract conservation, it is important to have microbiological
control of the method of dilution plant extracts in in vitro culture
media to avoid contamination and bias in the results.
This study therefore aims at testing the antifungal activity of P.
nitida extracts on plant fungi. More specifically, the study (i) deter-
mined the percentage yield of extracts (ethanol and aqueous) of P.
nitida fruit with climatic gradient; (ii) assessed thermo-stability of
P. nitida extracts; (iii) evaluated the antifungal potential of extracts
on F. oxysporum for the extraction and dilution types.
2. Material and methods
2.1. Vegetal material and fungal isolation
The vegetal material used was mainly P. nitida fruits (pulp and
seeds). The ripe fruits were directly collected from trees in two
contrasting climatic zones (Fig. 1). These were the Guinea-
Congolese region of West Africa (N 05°38.640 and W 04°05.017)
and Dahomey Gap zone (N 06°45.107 and E 02°42.697).
Isolates of F. oxysporum were obtained on the basis of isolates
made from tomato plants showing symptoms of wilt and crown
rot in the Phytopathology Laboratory of University Félix
HOUPHOUËT-BOIGNY of Cocody (Côte d’Ivoire) according to the
method described by Rivera-Vargas et al. (2006). Different parts
of the plant (roots, crowns and stems) were taken for isolation of
the pathogen. The samples were cut into fragments of approxi-
Ghislain Comlan Akabassi, Elie Antoine Padonou, Edouard Jacques Kouadio Yao et al.
mately 0.5 cm, then disinfected by soaking in sodium hypochlorite
solution (10%) for a period of 5 min. The fragments were then
washed with sterile distilled water and then dried between 2 ster-
ile filter papers. Once dried, the fragments were placed in Petri
dishes containing PDA (Potato Dextrose Agar) medium amended
with streptomycin sulfate at a rate of 200 mg/l in order to prevent
the proliferation of bacteria. Plates were incubated in the dark at
25 °C for 4–5 days. The purification of the pathogen was carried
out by successively transplanting fragments of the growth front
of the young developing cultures. In order to ensure the purity of
the colonies, single-pore cultures were applied for all isolates
obtained. The identification of the pathogen was carried out micro-
scopically, based on the characteristics of macroconidia, phialides
and chlamydospores (4), using the determination key presented
by Nelson et al. (1983).
2.2. Sampling and data collection
Three morphotypes of P. nitida have been identified in DG (dry
zone with low rainfall and high temperatures) and one in the GC
region of West Africa (humid region with high rainfall and low
temperature) (Akabassi et al., 2021). There are large rounded fruit
morphotypes (1), lengthened oval fruit morphotypes (2) and
rounded short fruit morphotypes (3) in DG and the large rounded
fruit morphotypes (1) in GC region. Because of penury of ripe fruits
from morphotypes 2 during the data collect period, only morpho-
types 1 and 3 were used in this study. To assess the yield of the
fruit populations according to climatic zone, the morphotypes 1
(M1D) and 3 (M3D) from DG zone and the morphotypes 1 (M1G)
from GC region were used. However, to assess the fungicide poten-
tial activities of extracts, only the morphotypes 1 (M1D) and 3
(M3D) from DG zone were used due to the limitation of laboratory
materials such as saccharose and agar. Two fruits of 800 g each
from each morphotype were rinsed with water to remove dirt
and cut separately into small pieces. They were then dried in an
oven at a temperature of 50 °C for 72 h and ground to powder using
a grinder.
2.3. Preparation of plant extracts
2.3.1. Preparation of total aqueous extract (TEA)
The preparation of the TEA was carried out according to the
method described by Zirihi et al. (2003). This method consists of
macerating 100 g of vegetable powder in 1L of distilled water.
The macerate is homogenised for 15 min using a multifunctional
blender sharpener at room temperature. The homogeneous solu-
tion obtained was filtered once with poplin, successively three
times on cotton wool and once on Whatman 3 mm filter paper.
The filtrate obtained was evaporated using a Med Center Venticell
oven at 50 °C for 72 h.
2.3.2. Preparation of the hydroethanolic extract 70 % (EE70%)
EE70% was obtained by macerating 100 g of vegetable powder
in 1L of 70% ethanol solution then homogenized for 15 min using
a Multifunctional Blender sharpener. The homogeneous solution
obtained was filtered once with poplin, successively three times
on cotton wool and once on Whatman 3 mm filter paper. The fil-
trate was evaporated using a Med Center Venticell oven at 50 °C
for 96 h.
2.4. Effect of dilution ways and extract types on the mycelia growth of
the mycopathogen
In order to identify the suitable dilution way and temperature
effect on the stability of the extracts, two dilution types were
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Journal of the Saudi Society of Agricultural Sciences 21 (2022) 425–431
tested; (1) dilution of the extracts in PDA medium before autoclav-
ing (AvA) and (2) dilution of the extracts in PDA medium after
autoclaving (ApA). Four extract types (2 TEA and 2 EE70%) from
DG were used at a concentration of 3 mg.ml
1. Fruit morphotypes
from GC region was used to compare the percentage yield accord-
ing to climate gradient.
Eight solutions of 40 ml were prepared by solubilizing them in
sterilized distilled water. From the eight solutions, 120 mg of each
of the four extracts were incorporated into four solutions before
autoclaving and the remaining four remained without extract.
The solutions were prepared by autoclaving at a temperature of
121 °C under a pressure of 1 bar for 2 h. After cooling the solutions
to 45 °C, the extracts were then incorporated into the other four
solutions at the same concentration. The PDA solutions with
extracts were homogenized and distributed in 50 mm diameter
Petri dishes, 10 ml per dish. Mycelial discs of 7 mm diameter, were
taken from 7-day old cultures with a punch and placed in the cen-
ter of the Petri dishes containing the PDA-extract mixture. All cul-
tures were incubated in a culture chamber at room temperature of
22 ± 2 °C for a 12-hour photoperiod. For each treatment, five Petri
dishes were used and the experiment was repeated three times.
Five controls without extract were carried out under the same cul-
ture conditions.
Radial mycelial growth of fungal colonies was assessed daily
until the surface of the culture medium was completely covered
in the control Petri dishes for 7 days. Measurements of radial
mycelial growth were made along the two perpendicular diagonals
drawn at the bottom of each petri dish with a cut-off point in the
middle of the mycelial disc.
The effect of the extracts on the growth of the fungus was deter-
mined for each method of dilution of the extracts of each morpho-
type of fruit from the inhibition rate (Ic) of the mycelial growth
according to the following formula (Nakpalo et al., 2018):
D0
DC
Ic ¼  100 ð1Þ
D0
where D0 is the mean diameter of the control colonies and Dc is the
mean diameter of colonies growth in the presence of the extracts
according to each dilution method.
2.5. Data analysis
The percentage yield of the extracts was determined according
to the formula,
MES
R¼  100 ð2Þ
MEE
where MES is the mass of the dry extract obtained and MEE is the
mass of the extracted sample. The effect of the dilution way, extrac-
tion types and the fruit morphotypes were analyzed using an
ANOVA with interactions. In case a significant difference was
observed, the means were compared using the Student-Newman-
Keuls test. No data transformation was applied on inhibition rate
data because normality and homoscedasticity were verified without
transformation using the Ryan-Joiner normality test and the Levene
variance homogeneity test. Analyzes were carried out using the Sta-
tistica version 7.1 software.
3. Results
3.1. Extracts of fruit morphotypes
In aqueous medium, there was no significant difference in yield
between the two fruit morphotypes from DG zone (M1D: 36.015%
and M3D: 36.211%) whereas there was significant difference
Ghislain Comlan Akabassi, Elie Antoine Padonou, Edouard Jacques Kouadio Yao et al. Journal of the Saudi Society of Agricultural Sciences 21 (2022) 425–431

Table 1 3.2. Effect of dilution way and antifungal potential of the extracts on
Percentage extract yield of fruit morphotypes from two climatic zones. Fusarium oxysporum
Dry extract yield (%)
Morphotypes of fruits collected TEA EE70% Results of the ANOVA with interactions (Table 2) performed on
inhibition rate of F. oxysporum according to the dilution way and
M1D 36.015 27.090
M3D 36.211 29.860
the extract types of the fruit morphotypes indicated that the factor
M1G 27.506 27.074 ‘‘Time” and some interactions (Morphotype  Extraction methods;
Time  Morphotype  Extraction method;
M1D: large rounded fruit morphotype from DG; M3D: rounded short fruit mor-
photype from DG; M1G: large rounded fruit morphotypes from the GC region; TEA:
Time  Morphotype  Dilution method) had a significant effect
Aqueous extract type; EE70%: ethanolic (70%) extract type. on inhibition rate of pathogen (p < 0.05). This means that the
extract type influenced the effectiveness of the antifungal potential
of P. nitida fruit morphotypes. However, the effect of the dilution
way was only noticeable over time.
40
The ethanolic extract type obtained the best inhibition rate
35 (42%) compared to the total aqueous extract type (35%) (Fig. 3;
30 Photo 1). The daily evolution of the inhibition rate of the two
Percentage (%)

25 extract types showed that the curve of the inhibition rate of the
20 ethanolic extract type was always above the aqueous extract types
TEA curve (Fig. 4).
15
EE70% The dilution way of the extracts after autoclaving (ApA)
10
obtained a inhibition rate of 41.58% while the dilution way of the
5 extracts before autoclaving (AvA) obtained a inhibition rate of
0 40.28% (Fig. 5). The daily evolution of the inhibition rate of the
M1D M3D M1G two dilution way showed that the inhibition rate curve of ApA
Fruit morphotypes always surpasses that of AvA (Fig. 4)
Fig. 2. Percentage yield of aqueous and ethanolic extracts of P. nitida fruit
morphotype; M1D: large rounded fruit morphotype from DG; M3D: rounded short
fruit morphotype from DG; M1G: large rounded fruit morphotype from the GC
region; TEA: Aqueous extract type; EE70%: ethanolic (70%) extract type.
50.0 b
45.0
(p < 0.001) in yield between the large rounded fruit morphotype 40.0 a
Inhibition rate (%)

from GC region (M1G) and those from DG zone (Table 1). Nearly 35.0
8.509 yield difference was observed between large rounded fruit 30.0
morphotype from DG zone and large rounded fruit morphotype 25.0
from GC region (M3D – M1G) and 8.705% difference yield differ- 20.0
ence was observed between rounded short fruit morphotypes from 15.0
DG zone and large rounded morphotype from GC region (M3D – 10.0
M1G) (Fig. 2). 5.0
In ethanolic medium, there was no significant difference in 0.0
yield between M1D (27.090%) from DG zone and M1G (27.074%) TEA EE70%
Extract types
from GC region. However, the extract yield of rounded short fruit
morphotypes from DG zone (M3D) was higher than that of large Fig. 3. Inhibition rate of F. oxysporum growth of in the extracts. TEA: Aqueous
rounded fruit morphotype from DG zone (M1D) with a 02.77% dif- extract type; EE70%: ethanolic (70%) extract type. The means followed by the same
letter are not significantly different (p  0.05 according to the SNK test).
ference (Fig. 2).

Table 2
ANOVA results on inhibition rate of F. oxysporum according to the dilution and extraction methods of the fruit morphotypes of P. nitida.

Source DF Type III SS Mean Square F Value

Morphotype 1 13.70 13.70 0.02 ns
Extract types 1 1084.87 1084.87 1.60 ns
Dilution way 1 331.17 331.17 0.49 ns
Morphotype  Extypes 1 6248.75 6248.75 9.20***
Morphotype  DiWay 1 1608.56 1608.56 2.37 ns
Extypes*DiWay 1 530.18 530.18 0.78 ns
Morphotype  Extypes  DiWay 1 105.21 105.21 0.15 ns
Time 6 14798.80 2466.47 29.49***
Time  Morphotype 6 138.23 23.04 0.28 ns
Time  Extypes 6 742.93 123.82 1.48 ns
Time  DiWay 6 112.44 18.74 0.22 ns
Time  Morphotype  Extypes 6 2017.84 336.31 4.02***
Time  Morphotype  DiWay 6 1244.45 207.41 2.48**
Time  Extypes  DiWay 6 346.51 57.75 0.69 ns
Time  Morphotype  Extypes  DiWay 6 13.29 2.22 0.03 ns

Extypes: extraction methods, DiWay: dilution ways, DF: degree of freedom, Type III SS: Sum of squares of deviations, F value: Fisher value, ns: not significant, ***: significant
at 1‰.

428
Ghislain Comlan Akabassi, Elie Antoine Padonou, Edouard Jacques Kouadio Yao et al. Journal of the Saudi Society of Agricultural Sciences 21 (2022) 425–431

45.0 4. Discussion
40.0
35.0 4.1. Extracts of fruit morphotypes
Inhibition rate (%)

30.0
25.0 The yield of extracts in the aqueous solution of DG fruit mor-
20.0 ETA photypes was 8% higher than that of fruit morphotypes from the
15.0 EE70%
GC region. The fruit morphotypes from DG region may be less rich
10.0 in water and more concentrate in active principles, which may be
5.0 due to the high temperature and relatively low rainfall (maximum
0.0 1400 mm/year) of the region. However, fruit morphotypes from
1 2 3 4 5 6 7 the GC region (with high rainfall, 3,000 mm/year) were rich in
Days water and less concentrate in active principles. These results cor-
roborate those of Péné (1999), who showed that sugar-cane in
Fig. 4. Trend of the F. oxysporum inhibition rate in the extracts. TEA: Aqueous southern Côte d’Ivoire (GC zone with high rainfall) was less con-
extract type; EE70%: ethanolic (70%) extract type.
centrated in sugar than that of northern Côte d’Ivoire (Sudanian
zone with low rainfall), which was more concentrated in sugar,
thus climate impacts the extracts yield. Rounded short fruit mor-
44.0 a photypes had higher yields compared to the large rounded mor-
photypes. In general, large fruits accumulate more water reserve
43.0 with a relatively low composition compared to short fruits that
a
42.0 are poor in water but rich in active principles.
Inhibition rate (%)

The fruit extract yield observed in this study is higher than that
41.0
observed in the leaves in both methanolic and aqueous solutions
40.0 (Dibua et al., 2013). These yields are much lower than those of
39.0 fruits whatever the climatic region of origin of the fruits. It can
therefore be concluded that fruits (pulp, seed) of P. nitida were
38.0
more concentrated in active principles.
37.0 The difference in yield observed between Total Aqueous Extract
36.0 (TEA) and 70% Ethanolic Extract (EE70%) could be explained by the
ApA AvA broad extraction spectrum of TEA. Water is a solvent that extracts a
Dilution methods large group of chemical compounds (small, medium and large
molecules) and EE70% concentrates many small and medium
Fig. 5. Inhibition rate of F. oxysporum growth according to the dilution way and active principles (terpenoids, phenols, polyphenols, quinones,
extract type in PDA medium. PDA: Potato Dextrose Agar; AvA: dilution way of
etc.). This could justify the higher yield obtained in TEA. These
extracts before autoclaving; ApA: dilution way of extracts after autoclaving; The
means followed by the same letter are not significantly different (p  0.05 results were in line with those obtained by Bene et al. (2015)
according to the SNK test). who explained that the yield of TEA from Harrisonia abyssinica
leaves was much higher than that of EE70% because of the wide
extraction spectrum in TEA.

TE

TEA EE70%

Photo 1. Photo showing the inhibition rate of Fusarium oxysporum on aqueous and ethanolic extracts; TE: Control box; TEA: Aqueous extract type; EE70%: ethanolic (70%)
extract type.

429
Ghislain Comlan Akabassi, Elie Antoine Padonou, Edouard Jacques Kouadio Yao et al.
4.2. Dilution way and antifungal potential of the extracts on Fusarium
oxysporum
The results of ANOVA with interaction showed that EE70%
extract type was more efficient (42% Inhibition Rate (IR)) compared
to the TEA extract type (40% IR). The low activity observed in TEA
could be attributed to the inability of water to extract more bioac-
tive compounds (due to its high oil content) readily available with
ethanol, which allowed more complete extraction, including less
polar compounds, many of which have antifungal properties. This
observation was also supported by Dibua et al. (2013) who showed
that the methanolic extract of P. nitida seeds had much better lar-
vicidal properties than the aqueous extract against Anopheles gam-
biae larvae. In addition, ethanol allowed a better concentration of
active ingredients compared to TEA (Okoli and Schabram, 2010;
Bagré et al., 2014; Bene et al., 2015).
No significant difference between the IRs of the two extract
dilution ways (p > 0.05). Both dilution ways were also valid. Con-
taminants could be eliminated during the AvA extract dilution
way. The extract can be qualified as thermostable if the heating
the extracts to 121 °C has the same efficiency as heating at the
extracts to 45 °C. This implies that the temperature had no effect
on the effectiveness of P. nitida active principles. This thermosta-
bility of the extracts could be explained by the high alkaloid con-
tent of P. nitida. Alkaloids are toxic and therapeutic substances
possessing the properties of alkalis (generic name for bases, sub-
stances which chemical properties are similar to those of soda
and potash).
As far as fruit morphotypes concerned, there was no significant
difference in their effects on the pathogen (p > 0.05). This showed
that morphotypes were subject to the same climatic conditions.
The only difference was the water level in the large round fruit
morphotype.
5. Conclusion
This study assessed the antifungal activity of two extract types
obtained from the Picralima nitida fruits. The total aqueous extract
type produced more extract than the ethanolic extract type. How-
ever, the aqueous extract was less potent in inhibition Fusarium
oxysporum than the ethanolic extract. P. nitida extracts were ther-
mostable. The temperature has no effect on the effectiveness of P.
nitida extracts. The short fruit morphotypes had the best inhibition
rate. This study offers the possibility to use P. nitida fruit extracts in
control strategies against Fusarium oxysporum.
Declaration of Competing Interest
The authors declare that they have no known competing finan-
cial interests or personal relationships that could have appeared
to influence the work reported in this paper.
Acknowledgements
Our honest thanks to ‘‘Centre d’Excellence Africain sur les
Changements Climatiques, la Biodiversité et l’Agriculture Durable
(CEA-CCBAD)” of Côte d’Ivoire which supported the cost related
to data collection. We are grateful to all the local communities of
Dahomey Gap and Guineo-Congolese region for helping during
the fieldwork.
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