Wood Metabolites of Myrcia insularis Gardner (Myrtaceae) have Potential Anti-Candida Activity

Abstract

The present work aimed to isolate secondary metabolites from Myrcia insularis Gardner (Myrtaceae) wood and to evaluate the anti-Candida activity and further extracts obtained by partition and the respective main isolated compounds. Wood was collected in a Seasonal Semideciduous Forest remnant of the Atlantic Forest of northern Rio de Janeiro State, Brazil. Chromatographic and spectrographic techniques were used to isolate and identify secondary metabolites. Methanol extract inhibited the growth of Candida buinensis and Candida tropicalis, with inhibition being approximately 82% for the latter. The main compound isolated from the ethyl acetate partitions was arjunolic acid, a triterpene. The antimicrobial activity was first observed with the wood metabolites of M. insularis adds to our understanding of the antifungal properties of this species and other species within the Myrtaceae family, including the presence of arjunolic acid, which may play a role in this activity.

Keywords:
Atlantic Forest; phytochemistry; triterpene; antifungal activity; arjunolic acid

1. INTRODUCTION AND OBJECTIVES

Studying the classes of compounds found in plants has contributed to discovering new natural compounds with applications in diverse areas, such as agriculture, medicine, and pharmacology (Vieira et al., 2016). More than 25% of all medicines are of plant origin, making them an important study subject. Besides, it´s necessary to search different sources to obtain active principles responsible for pharmacological actions and/or therapeutics (Ahmed et al., 2016; Gevú et al., 2019).

Plants possess a wide variety of secondary metabolites, alkaloids, terpenes, flavonoids, lignins, tannins, and glycosides (Bisoli et al., 2008; Vieira et al., 2016; Laursen et al., 2015; Knudsen et al., 2018; Li et al., 2020). Many of these compounds possess a pharmacological potential and can be found in all parts of the plant, including roots, leaves, flowers, and stems (Tungmunnithum et al., 2018).

The improper or indiscriminate use of antibiotics has contributed to the emergence of microorganisms with resistant multiple drugs (Chandra et al., 2017). This scenario has discovered new antimicrobial agents needed to combat these microorganisms, increasingly becoming one of the biggest threats to global health (Afroz et al., 2020). Since antiquity, natural products have performed essential roles worldwide in treating diseases of humans and other animals. They have also served as a source of new microbial agents due to their unique and enormous chemical diversity (Torrent et al., 2012; Wong et al., 2015; Ghosh et al., 2019; Afroz et al., 2020).

The occurrence of invasive fungal diseases in humans, such as candidiasis, is a concern for the global population, affecting immunosuppressed patients and elevating the number of deaths from septicemia (Scorzoni et al., 2017). There are about 200 species described for the fungal genus Candida, of which 17 have been identified as pathogenic to man. Although Candida albicans continues to be the most common clinical isolate, other species of Candida are being recovered from clinical samples with increasing frequency, such as C. glabrata, C. parapsilosis, C. tropicalis, and C. krusei, among others (Gabaldón et al., 2016; Colombo et al., 2017; Sharma et al., 2019). Infections caused by fungi of the Candida are usually treated with synthetic antifungal agents, especially azoles (itraconazole and fluconazole) and opolien (amphotericin B). However, these drugs have disadvantages, such as high toxicity to the host and the emergence of resistant strains (Duraipandiyan & Ignacimuthu, 2011; Spampinato & Leonardi, 2013; Perfect, 2017; Shekhova et al., 2017). The concern and studies of new drugs have grown in recent years in search relates natural products to an inhibitory activity against Candida spp. (Cavalcanti et al., 2011; Alexandrino et al., 2016; Gevú et al., 2019; Wang et al., 2021).

Speciesofthefamily Myrtaceaeareknownforproducingsecondary metabolites, emphasizing phenolic compounds. Investigations of somespecies havefound terpenes, flavonoids, steroids, andtannins (Araújo et al., 2019; Batiha et al., 2020). The genus Myrcia includes species with pharmacologic potential, including a hypotensive, diuretic, hypoglycemic, antidiarrheal, antimicrobial, antitumor, and hepatoprotective properties (Silva et al., 2019).

Some studies of metabolite isolation and biological activity with Myrcia spp. have revealed hypoglycemic activity for myricetin, compounds known as “insulin plant,” anti- inflammatory, antinociceptive, antioxidant, enzyme inhibitor, antifungal, antibacterial, or antimicrobial activities in general and acaricide were also described for Myrcia essential oils (Cerqueira et al., 2007; Cascaes et al., 2015; Ribeiro et al., 2022), mainly extracted from the leaves, flowers and fruits and less frequently on bark and fine stems. Terpenes have diverse biological proprieties, such as anticancer, antifungal, antiparasitic, antibacterial, antiallergic, antihyperglycemic, and immunomodulatory activities (Chan-Bacab & Pena-Rodrigues, 2001; Theis & Lerdau, 2003; Salem & Werbovetz, 2006; Paduch et al., 2007; Masoko et al., 2008, 2010; Liu, 2011; Zacchino et al., 2017; Wang et al., 2021). Studies with similar chemical classes evidenced activities relevant to the central nervous system, such as sedative, anticonvulsant, and pro-convulsive activities, and compounds used in natural insecticides (Passos et al., 2009; Huang & Osbourn, 2019) (Table 1). So, the Myrtaceae family, has great economic potential, with species used for food and pharmaceutical purposes. Even with this representativeness, the species Myrcia insularis Gardner (Myrtaceae), has no studies related to its wood, which is essential to contribute to research on anti-candida pharmacological effects.

Table 1
Main terpenoids found in amounts greater in Myrcia species and their chemical and biological activities. (Jorge et al., 2000; Limberger et al., 2004; Stefanello et al., 2006; 2010; Andrade et al., 2012; Silva et al., 2013; 2018; Cascaes et al., 2015; 2019; Pereira Junior, 2018; Santana et al., 2018; Silva et al., 2018; Franco et al., 2021; Jerônimo et al., 2021; Ribeiro et al., 2022; Fehlberg et al., 2023; Santana et al., 2023; Santos et al., 2023).

Species	Main Terpenoids	Chemical and biological activities
M. alagoensis O. Berg.	Germacrene B.	Antibacterial (Gram-positive and Gram-negative)
M. bella Cambess.	Guaiol acetate; a-cadinol; Sesquiterpene Hydrocarbons; Oxigenated sesquiterpenes.	Antioxidant, Allelopathic, Hypoglycemic, Antinociceptive
M. bracteata (L.C. Rich.) DC.	(E)-nerolidol; (E)-β - farnesene, spathulenol	Hypoglycemic, Antinociceptive, Antioxidant.
M. cuprea (O. Berg.) Kiaersk.	Myrcene; β-caryophyllene; α-pinene; germacrene D; Germacrene B.	Anticholinesterase; Larvicide; Fungicide
M. fallax (L.C. Rich.) DC.	α-pineno; β-pineno; β-elemene.	Antioxidant, Antiproliferative, Antimicrobial Activity, Antifungal
M. fenestrata DC.	(E)-Cariofileno; Espatulenol; α-Cadinol; Caryophyllene oxide; β-Copaen-4-α-ol.	Antibacterial, Antimalarial, Antioxidant, Anthelmintic
M. hiemalis Cambess.	2α,3β,21α-Trihydroxy-28,20β-hydroxytaraxastanolide; Sesqui-, di- and tetraterpenoids eudesm-4-(15)-en-7α,11-diol and geranylgeranyl acetate and α-tocopherol.	Antiparasitic, Antibacterial; Antifungal.
M. laruotteana Cambess.	α-bisabolol; 14-hidroxi-α-muuroleno.	Antioxidant, Antiproliferative Activity.
M. multiflora (Lam.) DC.	β-caryophyllene; Selin-11-en-4α-ol.	Antioxidant, Enzyme Inhibitor, Hepatoprotective, Antiobesity, Hypolipidemic, Allelopathic.
M. myrtillifolia DC.	Betulinic acid; Betulonic acid; Betulinaldehyde; Betulona; Oleanolic acid; ursolic acid.	Antimicrobial Activity.
M. oblongata DC.	Caryophyllene oxide; Trans-verbenol.	Antioxidant, Antimicrobial, Acaricidal.
M. obtecta Kiaersk.	Trans-calameneno; Monoterpeno α-terpineol.	Antioxidant.
M. pubiflora DC.	Cariofileno; Mustakone; 1,8-cineol; Tricicleno.	Antinociceptive, Anti-inflammatory
M. pubipetala Miq.,	Biciclogermacreno; Espatulenol.	Antienzymatic, Antioxidant.
M. rostrata DC.	Carotol; Germacreno B; E-cariofileno; Germacreno D; Dauceno.	Antibacterial, Antifungal
M. rufipila McVaugh.	β- and α-amyrin; Desmantin-I; and some flavonoids derived from terpenoids: Dihydromyricetin-4’-Ogallate; myricetin-3-O-α-L-rhamnopyranoside; Myricetin; Myricitrin; Isovitexin.	Hypoglycemic, Antioxidant.
M. splendens DC.	E-caryophyllene	Antinoceptive, Antifungal, Anti-inflammatory, Allelopathic, Insecticide. Antioxidant, Cytotoxic Activity against gastric, melanoma and colon human cancer cells.
M. sylvatica DC.	E-caryophyllene; γ-Elemene; Germacrene B.	Antioxidant Capacity, Cytotoxic Activity against melanoma and gastric human cancer cells.
M. tomentosa DC.	γ-elemeno; Germacreno D; (E)-cariofileno.	Antifungal, Antibacterial, Antioxidant, Allelopathic

Myrcia insularis, commonly called vapiranga or guapiranga, is endemic to Brazil, where it occurs in Espírito Santo, Rio de Janeiro, São Paulo, Paraná, Bahia, and Pernambuco (Santos et al., 2020). In Brazil, M. insularis is distributed in the Atlantic Forest domain in the vegetation of Dense Ombrophylous, Seasonal Semideciduous forest and Restinga, supposedly presenting fragmented subpopulations; it is found in several conservation units and has a high population density (CNCFlora, 2012). In northern Rio de Janeiro State, this species is known to occur in Seasonal Semideciduous Forest (SSF) in the Environmental Protection area of Morro do Itaoca, Campos dos Goytacazes, RJ, Brazil. As the species currently have little information on research related to the isolation of metabolites and biological activity they can be a potential source of research in the area, considering the family to which it belongs (Cascaes, 2015; 2019; Jeronimo et al., 2021). Thus, investigations of the phytochemistry of the wood of M. insularis can be necessary for studies about your biological proprieties, and contributes to knowledge about antifungal activities as a base for other species within Myrtaceae. So, this study aimed to isolate metabolites of M. insularis wood and evaluate the anti-Candida activity of the partitions. Beyond that, to isolate and identify the main compound of these partitions.

2. MATERIALS AND METHODS

2.1. Plant material and study area

Wood samples of M. insularis were collected by non- destructive methods of a remnant of SSF in the Atlantic Forest. The selected individuals (n. 5) without apparent defects, and with DBH above 10 cm had samples of the parts of the branches removed manually using a handsaw (Stanley®) and taken to the laboratory. The SSF remnant is located on a rocky outcrop of the Morro do Itaoca (21°48’0” S - 41°26’0” W) in the municipality of Campos dos Goytacazes, in northern Rio de Janeiro State (RJ), Brazil. The identified material was deposited in the wood collection “Xiloteca Drª Cecília Gonçalves da Costa” (HUENFw) under register nº 565, and herbarium HUENF of Universidade Estadual do Norte Fluminense Darcy Ribeiro (UENF) under register nº 10663 (Table 2). The samples were dried through the rotary oven and then ground by the hammer mill at the UENF Chemical Science Laboratory, resulting in approximately 2,0724 Kg.

Table 2
General data regarding of M. insularis on the Morro of Itaoca (SSF) and information on registration.

Data	SSF
Xiloteca HUENFw (nº)	565
Exsiccates HUENF (nº)	10663
Average height of individuals (m)	9.71
Average diameter of stalk (cm)	32.88

2.2. Isolation and extraction

The ground wood samples of M. insularis were subjected to extraction by cold maceration with methanol for four days. The extracts were concentrated in a rotary evaporator (FISATOM 802) and dried in a fume hood. This procedure was repeated three times, guaranteeing all the crude wood extract. A rotary evaporator was used to concentrate the extract, which was suspended in water and methanol (H₂O:MetOH - 1:3) and partitioned using the solvents dichloromethane (CH₂Cl₂), ethyl acetate (AcOEt), and butanol (ButOH) in ascending order of polarity (Figure 1). The steps were carried out at the UENF Chemical Sciences Laboratory.

Figure 1
Fractionation steps of the crude methanolic extract of M. insularis wood. collected at remnant of SSF in the Atlantic Forest.

The extracts obtained were submitted to anti-Candida activity tests, selects the best results of inhibition of yeast growth were selected for chromatographic and spectroscopic analysis.

2.3. Yeast growth-inhibition assay

Strains of the species Candida tropicalis (CE017) and Candida buinensis (3982) were maintained in Sabouraud agar (1 % peptone, 2 % glucose, and agar 1.7 %; Merck). The cell strains were provided by Physiology and Biochemistry of Microorganisms Laboratory at UENF, municipality of Campos dos Goytacazes, RJ, Brazil.

Inoculants of each stock of C. buinensis and C. tropicalis were transferred to Petri dishes containing Sabouraud agar and left to grow at 30 ºC for 24 h. Each cell aliquot was added to a 10 mL sterile culture medium (Sabouraud broth, 1 % peptone, 2 % glucose; Merck). The cells were counted in a Neubauer chamber (Optik Labor) under light microscopy (Axioplan, ZEISS). Only the CH₂Cl₂ and AcOEt extracts that presented significant antimicrobial activity were analyzed further in the other experiments.

The yeast strains C. buinensis, and C. tropicalis (1 x 104 cel.mL^-1) were incubated in Sabouraud broth containing 200, 100, and 50 μg.mL^-1 of each fraction of the CH2Cl2 and AcOEt extracts (solubilized in 20 % DMSO), with the final volume adjusted to 200 μL. The test was performed in 96-well microplates (NUNC, Nunclein Surface) at 30 ºC for 24 h. The control cells were cultured in the absence of extracts. The procedure followed the method of Broekaert et al. (1990) with adaptations and was performed entirely under aseptic conditions. Cell growth was analyzed after 24 h of incubation by optical density in a microplate reader at a wavelength of 620 nm. The cells were then observed by differential interference contrast (DIC) under a light microscope (Axioplan, A2, Zeiss). The experiments were performed in triplicate.

2.4. Spectrometry analysis

This analysis was performed by uni-two-dimensional Nuclear Magnetic Resonance (NMR) with the operation of 500 MHz for hydrogen (1H NMR) and 125 MHz for carbon-13 (¹³C NMR) with Ascend 500 (Brüker) spectrophotometer. Spectrometry analysis was performed with a high-resolution mass spectrometer (micrOTOF-Q II Brucker Daltonics) using the negative mode of study. Column chromatography (CC) was performed with a 60G silica gel (Merck), while thin layer chromatographic analysis was done with chrome aluminum foils (CCD Silica gel 60 F254, Merck). The compounds found were observed by ultraviolet irradiation at 254nm or 365nm and/or revealed with the sulfuric vanillin chromogenic reagent by warming.

The AcOEt (1.53g) partition was subjected to CC, eluted with CH₂Cl₂:MeOH of gradually increasing concentration until 100% of MetOH. Seven fractions (MIF1 - MIF7) were obtained. Fraction MIF4 (253mg) was resubmitted to CC, eluted with MetOH and CH₂Cl₂ until 100% of MetOH to obtain nine fractions (MIF4.1 - MIF4.9), with the compound 1 identified in the MIF4.6 (36.8 mg) fraction.

2.5. Statistical analysis

Growth inhibition data were evaluated by One-Way ANOVA, with differences of p < 0.05 being considered significant. All statistical analyses were performed using GraphPad Prism Software (6.0, version for Windows).

3. RESULTS

Of the three partitions obtained from the M. insularis wood only that of AcOEt and CH₂Cl₂ showed significance in the tests. The antimicrobial activity of the fractions obtained from AcOEt and CH₂Cl₂ was tested on the strains of C. buinensis and C. tropicalis using the concentrations of 200, 100, and 50 µg.mL^-1 (Figures 2 and 3).

Figure 2
The growth inhibition of yeast cells of C. buinensis (A) and C. tropicalis (B). Growth in the absence of extract (control) and the presence of 200, 100, and 50 μg.mL^-1 of AcOEt fractions. (*) Indicates significance by One-Way ANOVA (p < 0.05) among the treatments and their respective controls.

Figure 3
Growth inhibition of yeast cells of C. buinensis (A) and C. tropicalis (B). Growth in the absence of extract (control) and the presence of 200, 100, and 50 μg.mL^-1 of CH₂Cl₂ fractions. (*) Indicates significance by One-Way ANOVA (p < 0.05) among the treatments and their respective controls.

At concentrations of 200 and 100 µg.mL^-1, the AcOEt partition was able to significantly inhibit C. buinensis cells (Figure 2A), resulting in approximately 73% and 64% growth inhibition, respectively. The tested concentrations were also able to significantly inhibit C. tropicalis cells, with reductions of about 61%, 46%, and 15% in the growth of the strain in the concentrations of 200, 100, and 50 µg.mL^-1, respectively (Figure 2B).

The CH₂Cl₂ partition had a lower inhibitory effect against the yeasts than the AcOEt partition (Figures 2 and 3), with none of the three tested concentrations inhibiting the growth of C. buinensis cells (Figure 3A). However, growth inhibition against C. tropicalis cells was approximately 14%, 11%, and 8% for the concentrations of 200, 100, and 50 µg.mL^-1, respectively (Figure 3B). Data obtained through optical density corroborated the images obtained by light microscopy; however, only cells treated with 200 µg.mL^-1 of both partitions were recorded due to this concentration having the greatest inhibition (Figure 4).

Figure 4
Light microscopy of yeast cells of C. buinensis and C. tropicalis visualized by DIC after 24 h of incubation with 200 μg.mL^-1 extracts obtained from AcOEt or CH₂Cl₂. Bars: 20 μm.

Since the AcOEt partition inhibited more than 60% of yeast growth in the initial tests, it was selected for chromatographic analysis. The fraction MIF4 was set similarly for CC since it had better inhibition against C. buinensis. Finally, the fraction MIF4.6 showed growth inhibition of C. buinensis and C. tropicalis of approximately 72% and 82%, respectively (Figure 5).

Figure 5
Growth inhibition of yeast cells of C. buinensis (A) and C. tropicalis (B). Growth in the absence of extract (control) and in the presence of the MIF4.6 fraction.

The analysis of the MIF4.6 fraction by CCD revealed its degree of purity and, thus, was used to perform the NMR and HRESI- MS experiments. Based on these experiments, together with data from the literature (Table 3) and molecular structure (Figure 6), the substance was identified as arjunolic acid (Mann et al., 2012).

Table 3
₁H (500 MHz) and ₁₃C (125MHz) NMR data for compound 1, and correlation obtained by HSQC and HMBC spectra in CD₃OD as solvent.

C	δC	δH	HMBC
1	46.5	1.90 (m), 0.90 (m)	2, 25
2	68.3	3.71 (ddd, 11.3, 9.6, 4.5)	1, 3
3	76.8	3.37 (d, 9.6)	1, 2, 23, 24
4	42.7	-	3, 23, 24
5	46.8	1.65 (m)	1, 23, 24, 25
6	17.7	1.40 (m), 1.10 (m)	-
7	31.9	1.70 (m), 1.30 (m)	26
8	39.2	-	26, 27
9	47.5	0.95 (m)	12, 25, 26
10	37.6	-	1, 25
11	23.2	2.02 (m), 1.78 (m)	12
12	122.0	5.27 (t, 3.5)	18
13	144.0	-	12, 18, 27
14	41.6	-	18, 26, 27
15	27.4	1.80 (m), 1.08 (m)	27
16	22.6	2.02 (m), 1.78 (m)	-
17	46.3	-	16
18	41.3	2.84 (dd, 13.7, 4.1)	12
19	45.8	1.78 (m), 1.15 (m)	18, 29, 30
20	30.2	-	29, 30
21	33.5	1.75 (m), 1.40 (m)	-
22	31.4	1.56 (m), 1.25 (m)	-
23	64.9	3.52 (d, 11.1), 3.28 (d, 11.1)	3, 24
24	12.5	0.72 (s)	3, 23
25	16.1	1.05 (s)	-
26	16.4	0.84 (s)	-
27	25.1	1.20 (s)	-
28	180.4	-	16a, 18, 22a
29	32.2	0.93 (s)	30
30	22.6	0.96 (s)	29

Figure 6
Molecular structure of arjunolic acid extracted from the MIF4.6 fraction.

4. DISCUSSION

There have been important studies on plant secondary metabolism that produces compounds active in the defensive system. These metabolites have been extracted in different ways and from other plant parts and have served as the raw material for producing essential oils and extracts for different uses (Machado et al., 2006). These secondary metabolites perform various functions, such as protection against herbivores, pathogens, and other external influences, including temperature, humidity, and nutrient deficiency (Batiha et al., 2020).

Initial tests of the CH₂Cl₂, AcOEt, and ButOH partitions in the present study were performed to evaluate antifungal activity. The partitions with greater inhibition were those of CH₂Cl₂ and AcOEt. The AcOEt extract inhibited C. buinenis growth by about 73%. Giordani et al. (2015) found that CH₂Cl₂ and ethanol (C₂H₅OH) extracts of leaves and roots of Eucalyptus camaldulensis have positive anti-Candida actions. Despite the methanol extract from the wood having a lower inhibitory percentage, the data obtained in this study reiterate the presence of bioactive compounds against yeast.

The chromatography and spectrometry analyses of M. insularis extract isolated a skeleton triterpene pentacyclic arjunolic acid. This terpene was isolated from the MIF4.6 fraction derived from methanol extract. Arjunolic acid was previously isolated from leaves of other species of Myrtaceae, such as Melaleuca alternifolia (Vieira et al., 2004), Myrcia guianensis (Fehlberg, 2006), and Myrcia rotundifolia (Silva, 2014). Several studies have reported on the pharmacological potential of arjunolic acid. Facundo et al. (2005) considered this triterpene a multifunctional drug with anti-inflammatory, antinociceptive, and anticholinesterase activities. Besides that, this acid showed antifungal activity against Candida krusei, Candida albicans, and Cryptococcus neoformans (Silva et al., 2020).

Investigations of the chemical composition of the essential oil of the stem of Myrcia alternifolia identified diverse terpenes, including arjunolic acid, and another oil constituents presents antifungal, antibacterial, antiviral, and analgesic properties (Buck et al., 1994; Hammer et al., 1996; Halcón & Milkus, 2004; Veras et al., 2019). This triterpene isolated has shown a relatively high antimicrobial activity (Moreira, 2010). The author also mentioned that this substance was the major component (or the main component) found in M. alternifolia. On the other hand, several essential oil compounds showed that anti-Candida effectiveness was lower due to the interaction of other compounds present. The AcOEt and CH2Cl2 partitions of the present study inhibited growth by approximately 73% and 61% at 200 μg.mL^-1 for C. buinensis and C. tropicalis, respectively. So the fraction MIF4.6 inhibited about 72% and 82%, containing the isolated principal substance, arjunolic acid. Some terpenoids compounds function as repellents and/or attractants and are components of the typical aroma of several plant species, such as the high monoterpenes present in some essential oils (Silva et al., 2019; An et al., 2020). Furthermore, terpenes can be toxic at high concentrations and can serve as important combatants against pathogens and herbivory (Theis & Lerdau, 2003; Crowell, 2002).

There have been no published studies on the antimicrobial activity of M. insularis. Nevertheless, this study presents positive data regarding the growth inhibition potential of the species with the elimination of approximately 72% and 82% of C. buinensis and C. tropicalis, respectively, in 200 μg.mL-1 of the fraction MIF 4.6.

5. CONCLUSIONS

A compound belonging to the terpene class, arjunolic acid, was identified for the first time from M. insularis wood. The fraction contained this metabolic presented significant anti-Candida potential. Further investigations should be undertaken regarding the chemical and biological properties of this species of the family Myrtaceae, aiming for biotechnological applications, and serving as a basis for further studies.

REFERENCES

• Afroz M, Akter S, Ahmed A, Rouf R, Shilpi JA, Tiralongo E, Uddin SJ. Ethnobotany and antimicrobial peptides from plants of the solanaceae family: an update and future prospects. Frontiers in Pharmacology 2020; 11(565):1-15.
• Ahmed S, Hasan M, Mahmood Z. Antiurolithiatic plants: multidimensional pharmacology. Journal of Pharmacognosy and Phytochemistry 2016; 5(2):4-24.
• Alexandrino CR, Carvalho LP, Melo EJT, Mello EO, Gomes VM, Callado CH, Da Cunha M. Bioactivity of leaf extracts from species of Palicourea (Rubiaceae) on Trypanossoma cruzi, Candida sp and Fusarium solari. European Journal of Biomedical and Pharmaceutical Sciences 2016; 3(6):489-496.
• Al-Snafi A. Therapeutic properties of medicinal plants: a review of their dermatological effects (part 1). International Journal of Pharmacy Review & Research 2015; 5(4):328-337.
• An NTG, Huong LT, Satyal P, Tai TA, Dai DN, Hung NH et al. Mosquito larvicidal activity, antimicrobial activity, and chemical compositions of essential oils from four species of Myrtaceae from central Vietnam. Plants 2020; 9(544.):1-20.
• Andrade GS, Guimarães AG, Santana MT, Siqueira RS, Passos LO, Machado SMF, Adauto de S. Ribeiro AS, Sobral M, Almeida JRGS, Quintans-Júnior LJ. Phytochemical screening, antinociceptive and anti-inflammatory effects of the essential oil of Myrcia pubiflora in mice. Brazilian Journal of Pharmacognosy 2012; 22(1): 181-188.
• Araújo FF, Neri-Numa IA, Farias DP, Cunha GRMC, Pastore GM. Wild Brazilian species of Eugenia genera (Myrtaceae) as an innovation hotspots for food and pharmacological purposes. Food Research International 2019; 121:57-72.
• Batiha GE, Alkazmi LM, Wasef LG, Beshbishy AM, Nadwa EH, Rashwan EK. Syzygium aromaticum L. (Myrtaceae): Traditional uses, bioactive chemical constituents, pharmacological and toxicological activies. Biomolecules 2020; 10(202):1-16.
• Bisoli E, Garcez WS, Hamerski L, Tieppo C, Garcez FR. Bioactive pentacyclic triterpenes from the stems of Combretum laxum. Molecules 2008; 13(11): 2717-2728.
• Broekaert WF, Terras FR, Cammue BP, Vanderleyden J. An automated quantitative assay for fungal growth inhibition. FEMS Microbiology Letters 1990; 69(1-2):55-59.
• Buck DS, Nidorf DM, Addino JG. Comparison of two topical preparations for the treatment of onychomycosis: Melaleuca alternifolia (tea tree) oil and clotrimazole. The Journal of family practice 1994; 38(6):601-605.
• Cascaes MM, Guilhon GMSP, Andrade EHA, Zoghbi MGB, Santos LS. Constituents and pharmacological activies of Myrcia (Myrtaceae): a review of an aromatic and medicinal group of plants. International Journal Molecular Sciences 2015; 16(10):23881-23904.
• Cascaes MM, Guilhon GMSP, Zoghbi MGB, Andrade E. Flavonoids, antioxidant potential and antimicrobial activity of Myrcia rufipila McVaugh leaves (Myrtaceae). Natural Product Research 2019; 35(4): 1-5.
• Cavalcanti, YW, Almeida LFD, Padilha WWN. Screening of essential oils’ antifungal activity on candida strains. Odontologia Clínico- Científica (Online) 2011; 10(3):243-246.
• Cerqueira MD, Souza-Neta LC, Passos MGVM, Lima EO, Roque NF, Martins D et al. Seasonal variation and antimicrobial activity of Myrcia myrtifolia essential oils. Journal of the Brazilian Chemical Society 2007; 18(5):998-1003.
• Chan-Bacab MJ, Peña-Rodríguez LM. Plant natural products with leishmanicidal activity. Natural Product Reports 2001; 18(6):674-688.
• Chandra H, Bishnoi P, Yadav A, Patni B, Mishra AP, Nautiyal AR. Antimicrobial resistance and the alternative resources with special emphasis on plant-based antimicrobials-A. Plants 2017; 6(2):1-11.
• CNCFlora - Centro Nacional de Conservação da Flora. Myrcia insularis in Lista Vermelha da flora brasileira versão 2012.2 Centro Nacional de Conservação da Flora 2012. Available in <Available in http://cncflora.jbrj.gov.br/portal/pt-br/profile/Myrciainsularis >. Access: 05 jun. 2022.
  » http://cncflora.jbrj.gov.br/portal/pt-br/profile/Myrciainsularis
• Colombo AL, De Almeida Júnior JN, Slavin MA, Chen SC-A, Sorrell TC. Candida and invasive mould diseases in non-neutropenic critically ill patients and patients with hematological cancer. The Lancet Infectious Diseases 2017; 17(11):e344-e356.
• Crowell AL, Williams DC, Davis EM, Wildung MR, Croteau R. Molecular cloning and characterization of a new linalool synthase. Archives of Biochemistry and Biophysics 2002; 405(1):112-121.
• Duraipandiyan V, Ignacimuthu S. Antifungal activity of traditional medicinal plants from Tamil Nadu, India. Asian Pacific Journal of Tropical Biomedicine 2011; 1(2Suppplement):S204-S215.
• Facundo VA, Rios KA, Medeiros CM, Militão JSLT, Miranda ALP, Epifanio RA et al. Arjunolic acid in the ethanolic extract of Combretum leprosum root and its use as a potential multi-functional phytomedicine and drug for neurodegenerative disorders: anti-inflammatory and anticholinesterasic activities. Journal of the Brazilian Chemical Society 2005; 16(6B):1309-1312.
• Fehlberg I, Ferraz CG, Santos IBF, Santos IIP, Guedes MLS, Ribeiro PR, Cruz FG. A new C-methyl-flavone and other compounds from Myrcia guianensis. Biochemical Systematics and Ecology 2023; 106:104566.
• Franco CJP, Ferreira OO, Moraes AAB, Varela ELP, Nascimento LD, Percário S, Oliveira MS, Andrade EHA. Chemical composition and antioxidant activity of essential oils from Eugenia patrisii Vahl, E. punicifolia (Kunth) DC., and Myrcia tomentosa (Aubl.) DC., leaf of family Myrtaceae. Molecules 2021; 26(11):1-12.
• Gabaldón T, Naranjo-Ortíz MA, Marcet-Houben M. Evolutionary genomics of yeast pathogens in the Saccharomycotina. FEMS Yeast Research 2016; 16(6):1-10.
• Gevú KV, Lima HRP, Neves IA, Mello EO, Bonan GT, Carvalho LP et al. Chemical composition and anti-Candida and anti-Trypanosoma cruzi activities of essential oils from the rhizomes and leaves of Brazilian species of Renealmia L. fil. Records of Natural Products 2019; 13(3):268-280.
• Ghosh C, Sarkar P, Issa R, Haldar J. Alternatives to conventional antibiotics in the Era of antimicrobial resistance. Trends in Microbiology 2019; 27(4):323-338.
• Giordani C, Santin R, Cleff MB. Levantamento de extratos vegetais com ação anti-Candida no período de 2005-2013. Revista Brasileira de Plantas Medicinais 2015; 17(1): 175-185.
• Halcón L, Milkus K. Staphylococcus aureus and wounds: A review of tea tree oil as a promising antimicrobial. American Journal of Infection Control 2004; 32(7):402-408.
• Hammer KA, Carson CF, Riley TV. Susceptibility of transient and commensal skin flora to the essential oil of Melaleuca alternifolia (tea tree oil). American Journal of Infection Control 1996; 24(3):186-189.
• Huang AC, Osbourn A. Plant terpenes that mediate below‐ground interactions: prospects for bioengineering terpenoids for plant protection. Pest Management Science 2019; 75(9):2368-2377.
• Jeronimo LB, Costa JS, Pinto LC, Montenegro RC, Setzer WN, Mourão RHV, Silva JKR, Maia JGS, Figueiredo PLB. Antioxidant and cytotoxic activities of Myrtaceae essential oil rich in terpenoids from Brazil. Natural Product Communications 2021; 16(2): 1-13.
• Jorge LIF, Aguiar JPL, Silva MLP. Foliar anatomy of pedra-hume- caa (Myrcia sphaerocarpa, Myrcia guianensis, Eugenia punicifolia, Myrtaceae). Acta Amazônica 2000; 30(1): 49-57.
• Pereira Júnior RC. Caracterização química e avaliação dos potenciais antimicrobiano, inseticida e citotóxico de óleos essenciais obtidos de Myrcia spp. (Myrtaceae) ocorrentes em ecossistema de terra firme (Amazônia) [tese]. Manaus: Universidade Federal do Amazonas; 2018.
• Knudsen C, Gallage NJ, Hansen CC, Møller BL, Laursen T. Dynamic metabolic solutions to the sessile life style of plants. Natural Product Reports 2018; 35(11):1140-1155.
• Laursen T, Møller BL, Bassard JE. Plasticity of specialized metabolism as mediated by dynamic metabolons. Trends in Plant Science 2015; 20(1):20-32.
• Li Y, Kong D, Fu Y, Sussman MR, Wu H. The effect of developmental and environmental factors on secondary metabolites in medicinal plants. Plant Physiology and Biochemistry 2020; 148:80-89.
• Limberger RP, Sobral M, Henriques AT, Menut C, Bessiere JM. Essential oils from Myrcia species native to Rio Grande do Sul. New Chemical 2004; 27(6): 916-919.
• Liu WJH. Traditional herbal medicine research methods: identification, analysis, bioassay, and pharmaceutical and clinical studies. 3rd. New Jersey: John Wiley & Sons; 2011.
• Machado FAPSA, Capelasso M, Oliveira AJB, Zamuner MLM, Mangolin CA, Machado MFPS. Alkaloid production and isozymes expression from cell suspension culture of Cereus peruvianus Mill. (Cactaceae). Journal of Plant Science 2006; 1(4):324-331.
• Mann A, Ibrahin K, Oyewale AO, Amupitan JO, Fatope MO, Okogun JI. Isolation and elucidation of three triterpenoids and its antimycobacterial activity of Terminalia avicennioides. American Journal of Organic Chemistry 2012; 2(2): 14-20.
• Masoko P, Mdee LK, Mampuru LJ, Eloff JN. Biological activity of two related triterpenes isolated from leaves of Combretum nelsonii (Combretaceae). Natural Product Research 2008; 22(12):1074-1084.
• Masoko P, Picard J, Howard RL, Mampuru LJ, Eloff JN. In vivo antifungal effect of Combretum and Terminalia species extracts on skin wound healing in immunosuppressed rats. Pharmaceutical Biology 2010; 48(6):621-632.
• Moreira TMS. Estudo da composição química, citotoxicidade e alvos da atividade antifúngica de Melaleuca alternifolia Cheel (Myrtaceae) e de Plinia cauliflora (Mart.) Kausel (Myrtaceae) [dissertação]. Araraquara: Faculdade de Ciências Farmacêuticas Universidade Estadual Paulista; 2010.
• Paduch R, Kandefer-Szerszen M, Trytek M, Fiedurek J. Terpenes: compounds useful in human healthcare. Archivum Immunologiae et Therapiae Experimentalis 2007; 55:315-327.
• Passos CS, Arbo MD, Rates SMK, Von-Poser GL. Terpenoids with activity in the Central Nervous System (CNS). Brazilian Journal of Pharmacognosy 2009; 19(1A):140-149.
• Perfect JR. The antifungal pipeline: a reality check. Nature Reviews Drug Discovery 2017; 16(9):603-616.
• Ribeiro CL, Paula JAM, Peixoto JC. Pharmacological properties of species of the genres: Myrcia, Eugenia and Psidium-Myrtaceae-, typical of the Cerrado: A scope review. Research, Society and Development, 2022; 11(8): e44711830356.
• Salem MM, Werbovetz KA. Natural products from plants as drugs candidates and lead compounds against Leishmaniasis and Trypanosomiasis. Current Medicinal Chemistry 2006; 13(21): 2571-2598.
• Santana CB, Souza JGL, Coracini MDA, Walerius AH, Soares VD, Costa WF, Pinto FGS. Chemical composition of essential oil from Myrcia oblongata DC and potencial antimicrobial, antioxidant and acaricidal activity against Dermanyssus gallinae (DEGEER, 1778). Bioscience Journal 2018; 34(4): 996-1009.
• Santos C, Melo NC, Ruiz ALTG, Floglio MA. Antiproliferative activity from five Myrtaceae essential oils. Research, Society and Development 2023; 12(3): e14612340536.
• Santos MF, Amorim BS, Burton GP, Fernandes T, Gaem PH, Lourenço ARL et al. Myrcia in flora and Funga of Brazil. Botanical Garden of Rio de Janeiro. 2020. [dataset]. 5 Jun. 2022. http://floradobrasil.jbrj.gov.br/FB10709
  » http://floradobrasil.jbrj.gov.br/FB10709
• Scorzoni L, Silva ACAP, Marcos CM, Assato PA, Melo WCMA, Oliveira HC et al. Antifungal therapy: new advances in the understanding and treatment of mycosis. Frontiers in Microbiology 2017; 08:1-23.
• Sharma J, Rosiana S, Razzaq I, Shapiro RS. Linking cellular morphogenesis with antifungal treatment and susceptibility in Candida pathogens. Journal of Fungi 2019; 5(1):1-28.
• Shekhova E, Kniemeyer O, Brakhage AA. Induction of mitochondrial reactive oxygen species production by itraconazole, terbinafine and amphotericin B as a mode of action against Aspergillusfumigatus. Antimicrobial Agents and Chemotherapy 2017; 61(11):e00978-17.
• Silva EAJ, Estevam EBB, Silva TS, Nicolella HD, Furtado RA, Alves CCF et al. Antibacterial and antiproliferative activities of the essential oil of fresh leaves of Psidium guajava L. (Myrtaceae) Brazilian Journal of Biology 2019; 79(04):697-702.
• Silva FS, Landell MF, Paulino GBV, Coutinho HDM, Albuquerque UP. Antifungal activity of selected plant extracts based on an ethnodirected study. Acta Botanica Brasilica 2020; 34(2):442-448.
• Silva NA, Bomfim HF, Magalhães AO, Rocha ML, Lucchese AM. Chemical composition and antinociceptive activity of essential oil from Myrcia rostrata DC. (Myrtaceae) in animal models. New Chemistry 2018; 41(9):982-988.
• Silva NA, Uetanabaro ANT, Lucchese AM. Chemical composition and bacterial activity of essential oils from Myrcia alagoensis (Myrtaceae). Natural Product Communications 2013; 8(2): 269-271.
• Spampinato C, Leonardi DC. Infections, Causes, Targets and Resistance Mechanisms: Traditional and Alternative Antifungal Agents. BioMed Research International 2013; 2013(12):1-13.
• Stefanello MEA, Cervi AC, Wisniewski Júnior A, Simionatto EL. Essential oil composition of Myrcia laruotteana Camb. Journal of essential oil research 2006; 19: 466-467.
• Stefanello MEA, Cervi AC, Wisniewski Júnior A, Simionatto EL Composition and seasonal variation of essential oils of Myrcia obtecta (O. Berg) Kiaersk. var. obtecta, Myrtaceae. Brazilian Journal of Pharmacognosy 2010; 20(1):82-86.
• Theis N, Lerdau M. The evolution of function in plant secondary metabolites. International Journal of Plant Sciences 2003; 164(S3):S93-S102.
• Torrent M, Pulido D, Rivas L, Andreu D. Antimicrobial Peptide Action on Parasites. Current Drug Targets 2012; 13(9):1138-1147.
• Tungmunnithum D, Thongboonyou A, Pholboon A, Yangsabai A. Flavonoids and Other Phenolic Compounds from Medicinal Plants for Pharmaceutical and Medical Aspects: An Overview. Medicines (Basel) 2018; 5(3):1-16.
• Veras BO, Santos YQ, Oliveira FGS, Almeida JRGS, Silva AG, Correia MTS et al. Algrizea Minor Sobral, Faria & Proença (Myrteae, Myrtaceae): Chemical composition, antinociceptive, antimicrobial and antioxidant activity of essential oil. Natural Product Research 2019; 34(20):3013-3017.
• Vieira MGC, Santos FR, Braz Filho R, Vieira IJC. Chemical constituents of Trichilia hirta (Meliaceae). Natural Product Communications 2016; 11(5):593-596.
• Vieira TR, Barbosa LCA, Maltha CRA, Paula VF, Nascimento EA. Chemical constituents from Melaleuca alternifólia (Myrtaceae). Química Nova 2004; 27(4):536-539.
• Wang Y, Lu C, Zhao X, Wang D, Liu Y, Sun S. Antifungal activity and potential mechanism of Asiatic acid alone and in combination with fluconazole against Candida albicans. Biomedicine & Pharmacotherapy 2021; 139(4):1-7.
• Wong KY, Vikram P, Chiruvella KK, Mohammed A. Phytochemical screening and antimicrobial potentials of Borreria sps (Rubiaceae). Journal of King Saud University - Science 2015; 27(4):302-311.
• Zacchino SA, Butassi E, Libert MD, Raimondi M, Postigo A, Sortino M. Plant phenolic and terpenoids as adjuvants of antibacterial and antifungal drugs. Phytomedicine 2017; 37:27-48.

Edited by

Associate editor: José Luis Louzada https://orcid.org/0000-0002-0991-1711

Publication Dates

• Publication in this collection
  04 Dec 2023
• Date of issue
  2023

History

• Received
  03 Aug 2022
• Accepted
  19 Sept 2023