Fungicidal Activity Evaluation of Malunggay (Moringa oleifera) Leaf and Seeds

Methanolic Extract Against Aspergillosis (Aspergillus fumigatus)

CHAPTER 1

Introduction

The Philippines is a low to middle-income country, a tropical country in Southeast

Asia. Infectious diseases in the country remain the leading cause of the morbidity of the

population (M Batac, et al. 2017). Aspergillus species are globally pervasive saprophytes

found in a variety of ecological niches. Species of aspergilli have almost 200 species that

have been identified. Midst them, Aspergillus fumigatus is the most prevalent, and it is

the one that is in charge for the increased incidence of invasive aspergillosis (IA) (Taylor

R. T. Dagenais and Nancy P. Keller, 2020). Aspergillosis, which Aspergillus fumigatus

causes, is one of the common fungal infections in the Philippines. The devious fungal

pathogen is responsible for various human respiratory diseases, such as allergic

bronchopulmonary aspergillosis, aspergilloma, and invasive aspergillosis (Sophie Paris et

al. 2020). As stated by Jean-Paul Latgé and Georgios Chamilos (2019), aspergillosis

remains a significant problem on human health, with a quickly evolving human

epidemiology and a new group of at-risk patients. Immunocompromised patients cause a

usually fatal invasive aspergillus (IA).

Since fungicides are exceptionally costly and cause genuine environmental

pollution, control techniques are coordinated to supplant hazardous chemical fungicides

by naturally inviting characteristic items (Mamdouh and Eweis, 2007). Plant extracts are

often used as a safer alternative for synthetic drugs due to the latter’s high cost and
various side effects. There are immensely herbal plants in the Philippines alone that have

substantially anti-bacterial,anti-viral, antimicrobial properties.

Moringa oleifera, coming from the Moringaceae family, a plant commonly

known as the “drumstick” or “horseradish” tree. It is widespread in the Philippines,

including Asia, South Africa, Northeast Africa, and Northwest India (Meireles & Gomes,

2020). Height may vary between 10-12 m with a diameter of 45 cm; in terms of color, it

may appear pale gray or tan (Olson, 2010). Different parts of the said plant contains

antimicrobial properties and can be utilized in medicinal field and for other purposes.

Ascorbic acid, fatty acids and phenolic acids are several antioxidant compounds it

contains (Vongsak et al., 2014; Alhakmani et al., 2013).

Based on current findings, the other compounds in Moringa oleifera comprise

quercetin, zeatin, kaempferol, β-sitosterol, and caffeoylquinic acid (Sharma and Paliwal,

2013) along with myricetin, chlorogenic acid, rutin, glycosides niaziminin and niazin

(Mbikay, 2012). Other studies assessed that these compounds present are possessing

antimicrobial properties (Arora et al., 2014; Arora et al., 2013; Dwivedi and Enespa,

2012), antioxidant (Verma et al., 2009), antibacterial, antiulcer (Arora and Onsare,

2014; Belay and Sisay, 2014), anti-inflammatory, diuretic (Krishnamurthy et al., 2015)

anticancer (Pinto et al., 2015) and antifungal (Batista et al., 2014; Kadhim and Al-

Shammaa, 2014) .There are various of available reports in regards to bio-fungicide

efficacy of M. oleifera plant extracts against pathogenic fungi. Hence, further

investigations are required for deeper understanding of its fungicidal activity. Therefore,

in this study, the antifungal activity of M. oleifera’s methanolic leaf and seeds extract

will be eveluated on Aspergillus fumigatus.

Statement of the Problem

Generally, this study aims to “Evaluate the Fungicidal Activity of Malunggay

(Moringa oleifera) Leaf and Seeds Methanolic Extract Against Aspergillosis (Aspergillus

fumigatus)”

Specifically, this seeks answers to the following questions:

1. Does the methanolic leaf extract of M. oleifera has a fungicidal activity against

Aspergillus fumigatus ?

2. Does the methanolic seeds extract of M. oleifera has a fungicidal activity

against Aspergillus fumigatus ?

3. Is there a significant difference between the efficacy of the antifungal properties

of M. oleifera and the synthetic/chemical fungicides?

Hypotheses

1. The methanolic leaf extract of M. oleifera has a fungicidal activity against

Aspergillus fumigatus.

2. The methanolic seeds extract of M. oleifera has a fungicidal activity against

Aspergillus fumigatus.

3. There is no significant difference between the efficacy of the antifungal

properties of M. oleifera and the synthetic/chemical fungicides.

Significance of the Study

The findings of the study will be beneficial to the following:

Community. This study is all about the antifungal activity of M. oleifera on A.

fumigatus, which will be an absolutely significant finding to help prevent the spread of

the disease in the community; thus, people can have their low-cost yet safe medication.

Researchers. They will gain more information about the focus study and develop

new findings based on it. They can also contribute to the community in the field of

medicine.

Agricultural Suppliers and Farmers. The findings may provide knowledge for

the agricultural suppliers who supply M. oleifera seeds and for farmers who harvest the

said plant to increase their yield and gain more profit since it can be used for medicinal

purposes.

Future Researches. This study can be a source of information for future

researchers, especially those that are not widely discussed yet.

Chapter II
 Review of Related Literature and Studies

A. Related Literatures

Aspergillus fumigatus

Aspergillus fumigatus is one of the most significant common saprophytic

fungi found in the air. This fungus' conidia are regularly inhaled by humans and

animals (Jean-Paul Latgé 2020). Significant genome comparisons have shown

that no gene sets are shared entirely by both Aspergillus fumigatus and any other

human pathogen sequenced to date, such as Candida or Cryptococcus species,

Aspergillus fumigatus.

The enzymatic machinery compulsory by a fungus to take over plant

substrates has been found in the A. fumigatus genome ( Fredj Tekaia and Jean-

Paul Latgé 2005). The immune response of the host must be able to differentiate

between the various morphological forms of A. fumigatus to frustrate tissue

invasion and control fungal growth. At the same time, the fungus requires

nutrients and must acclimate to the hostile environment by dodging immune

recognition and counteracting host responses. A. fumigatus can adjust and

become resilient to antifungal drugs, highlighting the importance of designing

new antifungal drugs and strategies like immunotherapy ( Frank L. van de

Veerdonk et al., 2017).

Moringa oleifera
 Moringa oleifera, commonly known as Malunggay is a tiny, densely

branched tree that can reach a height of 9 meters and has a soft, white wood and

corky, sticky bark. Leaves are alternating and 25 to 50 cm long, frequently thrice

pinnate. On a compound (3 times pinnate) stalk, each compound leaf has 3-9 very

thin leaflets distributed. The leaflets are 1 to 2 centimeters in length, thin, ovate to

elliptic, and ovate to elliptic. Flowers on spreading panicles are white and fragrant,

about 1.5 to 2 cm in length. Pendulous, three-angled, and nine-ribbed pod is 15 to

30 centimeters long. Seeds have three angles, and the angles are winged.

(Stuartxchange, 2017).

Through research, it was found out that M. oleifera contains many

essential nutrients, like vitamins, minerals, amino acids, beta-carotene,

antioxidants, anti- inflammatory nutrients as well as as omega 3 and 6 fatty acids

(Fahey, 2005; Hsu et. al., 2006; Kasolo et. al., 2010). The plant has the presence

of several sorts of antioxidant compounds such as flavonoids, ascorbic acid,

carotenoids,and phenolics (Dillard & German, 2000; Siddhuraju & Becker, 2003).

Nutrition content of plant plays an essential function in medicinal, nutritional, and

therapeutic properties (Al-Kharusi et al., 2009).

Phytochemical composition of Moringa oleifera (MO)

Different portions of the MO tree have been shown to contain distinct

glucosinolates, flavonoids, and phenolic acids (Amaglo et al. 2010; Coppin et

al. 2013) Carotenoids (Saini et al. 2014c), polyunsaturated fatty acids (PUFAs)

(Saini et al. 2014d), tocopherols (Saini et al. 2014e), highly bioavailable minerals
(Saini et al. 2014a), and folate (Saini et al. 2014a) are all examples of highly

bioavailable minerals (Saini et al. 2016).