Asian Pacific Journal of Tropical Medicine 2017; 10(4): 351–360 351

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Asian Pacific Journal of Tropical Medicine

journal homepage: http://ees.elsevier.com/apjtm

Review http://dx.doi.org/10.1016/j.apjtm.2017.03.021

Acacia karroo Hayne: Ethnomedicinal uses, phytochemistry and pharmacology of an important

medicinal plant in southern Africa

Alfred Maroyi✉
Medicinal Plants and Economic Development (MPED) Research Center, Department of Botany, University of Fort Hare, Private Bag X1314, Alice
5700, South Africa

A R TI C L E I N F O ABSTRACT

Article history: Acacia karroo (A. karroo) has been used as herbal medicine by the indigenous people of
Received 16 Jan 2017 southern Africa for several centuries. The potential of A. karroo as herbal medicine, its
Received in revised form 15 Feb 2017 associated phytochemistry and biological activities are reviewed. The extensive literature
Accepted 14 Mar 2017 survey revealed that A. karroo is traditionally used to treat or manage 32 and five human
Available online 6 Apr 2017 and animal diseases and ailments, respectively. The species is used as herbal medicine for
diseases and ailments such as colds, diarrhoea, dysentery, flu, malaria, sexually trans-
mitted infections (STIs), wounds, and also as colic and ethnoveterinary medicine. Mul-
Keywords:
tiple classes of phytochemicals such as flavonoids, phenols, phytosterols,
Acacia karroo
proanthocyanidin, tannin, terpenes as well as several minerals have been identified from
Ethnomedicinal uses
leaves and roots of A. karroo. Scientific studies on A. karroo indicate that it has a wide
Ethnopharmacology
range of pharmacological activities which include antibacterial, antifungal, anti-
Ethnoveterinary medicine
gonococcal, antihelmintic, antilisterial, antimalarial, antimycobacterial, antioxidant, HIV-
Southern Africa
1 reverse transcriptase, anti-inflammatory and analgesic. A. karroo has a lot of potential as
Traditional knowledge
a possible source of pharmaceutical products for the treatment of a wide range of both
human and animal diseases and ailments. Future research should focus on the mecha-
nisms of action of the different plant parts used as herbal medicines, isolated compounds,
their efficacy, toxicity and clinical relevance.

1. Introduction therefore, renamed Vachellia karroo (V. karroo, Hayne) Banfi

& Galasso when the genus Acacia was renamed Vachellia.
Acacia karroo (A. karroo) Hayne is a member of the genus Taxonomically, Vachellia is closer to Senegalia, the main
Acacia Miller, family Fabaceae and subfamily Mimosoideae. difference is that Vachellia has capitate inflorescences (round,
The genus was first described by Philip Miller in 1754, the name head-like flowers) and spinescent stipules (thorns) while Sene-
was derived from the Greek word ‘akis’ which means point or galia has spicate inflorescences (flowers in spikes) and the
barb, referring to the thorns found on African Acacia species [1]. stipules are non-spinescent [3]. But taxonomists worldwide want
The species name ‘karroo’ is the old spelling for the South the name Acacia to be conserved as renaming the genus as
African semi-desert natural biome ‘karroo’, where the species Vachellia will create numerous taxonomic and retypification
was first described by botanical explorers [2]. The genus contains problems [5]. In literature both names are used, for example
a large number of species (approximately 1500), making it the V. karroo instead of A. karroo was used by Taylor and Barker
largest genus within the Fabaceae family and is widespread, [6] and Idamokoro et al [7]. But at the present moment
occurring in Australia, Asia, Africa and the Americas [3]. The V. karroo is regarded as an invalid name by the Royal Botanic
genus Acacia was re-classified recently into five distinct Garden and Missouri Botanic Garden plant name database
genera, Vachellia, Senegalia, Mariosousa, Acaciella and Acacia (www.theplantlist.org) and therefore, A. karroo has been
which are clearly distinct based on a number of morphological, adopted in this study.
anatomical and biochemical attributes [4]. A. karroo was A. karroo has been recorded throughout southern Africa,
ranging from the south-western Cape in South Africa, northwards
✉
First and corresponding author: Alfred Maroyi, Department of Botany, Uni- into Lesotho, Swaziland, Namibia, Angola, Botswana, Malawi,
versity of Fort Hare, Private Bag X1314, Alice 5700, South Africa. Mozambique, Zambia and Zimbabwe [6]. It has been introduced
E-mail: [email protected] to North Africa, Australia, India, Myanmar and South America
Peer review under responsibility of Hainan Medical University.

1995-7645/Copyright © 2017 Hainan Medical University. Production and hosting by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://
creativecommons.org/licenses/by-nc-nd/4.0/).
352 Alfred Maroyi/Asian Pacific Journal of Tropical Medicine 2017; 10(4): 351–360

(Argentina, Bolivia, Chile and Paraguay), where it is often used ethnobotany, ethnomedicinal uses, ethnopharmacology, phar-
as live fence around agricultural fields [8,9]. A. karroo grows macology, phytochemistry and therapeutic value) and
from sea level to 1800 m on soils ranging from pure geographical coverage (i.e., southern Africa). The following
unconsolidated sand to heavy clays with an annual rainfall keywords were used to search literature sources: A. karroo and
from 1500 mm down to less than 200 mm where ground water V. karroo. Other literature sources included papers published in
is available along drainage lines and around pans and dams [8]. international journals, reports from international, regional and
The species can grow under different climatic conditions but its national organizations, conference papers, books, theses, web-
limiting factors are water availability and intense cold [8]. A. sites and other grey literature. References were also identified by
karroo is the most widespread Acacia in southern Africa, searching the library collections of the National Herbarium and
occupying a diverse range of habitats including dry thornveld, Botanic Gardens (SRGH), Harare, Zimbabwe and the University
river valley scrub, bushveld, woodland, grassland, river banks of Fort Hare, South Africa.
and coastal dunes [6]. A. karroo shows a huge variety in terms
of its growth form, with plants from different areas in the 3. Species description and ethnomedicinal uses
species' geographical range often having a different appearance.
In the formal taxonomic revision of the species, Ross [10] A. karroo varies from a multi-stemmed shrub to a tree of up
detailed the vast range in morphology in this species, to 15 m in height [15]. The stem of A. karroo is dark brown to
describing seven different informal taxonomic entities of the almost black characterized by rough and somewhat flaky,
species which were generally correlated to its distribution. revealing reddish underbark [8]. A. karroo has pairs of large
Recently, Taylor and Barker [6] evaluated the genetic variability white spines which occur on the twigs and branches. The
of the species throughout South Africa using the Inter-Simple leaves comprise about five pairs of leaflets, each divided into
Sequence Repeat (ISSR) DNA ‘fingerprinting’ to determine ten or more pairs of smaller leaflets of about 5 mm long [16].
whether there is any genetic structure that correlates to the The branches bear minute golden-yellow, ball-shaped flowers
morphological diversity of the species. The authors concluded and the fruit is a long, narrow, spirally twisted pod [8].
that A. karroo should be considered as an ochlospecies, as the A. karroo boasts a large number of recorded ethnomedicinal
evolution of the observed morphotypes has been recent and rapid, and traditional uses in southern Africa (Table 1). The roots of
and therefore the genetic variation observed represents the A. karroo are used as remedy for colic in infants in Lesotho
ancestral gene pool that has not yet undergone lineage sorting as [17,18] and South Africa [19,20] while bark, gum and leaf infusions
a consequence of isolation. are used as remedy for diarrhoea and dysentery in South Africa
According to Barnes et al [8], A. karroo is a multipurpose tree [19–23] and Zimbabwe [24]. A. karroo is also widely used as
with great potential for increasing productivity in agroforestry herbal medicine for sexually transmitted infections (STIs) such
and silvopastoral systems over a wide range of sites in the dry as gonorrhoea and syphilis in Zimbabwe [25,26], sexually
zones of the tropics and subtropics. It is also categorized as a transmitted diseases and venereal diseases in South Africa
species with potential commercial value in Botswana, South [20,27]. The bark, gum and leaves are used as emollient and
Africa and Zimbabwe [8,11,12]. In Botswana, A. karroo exudate astringent for colds, conjunctivitis and haemorrhage [28]. In
is used for pharmaceutical purposes and is considered to be of Zimbabwe, roots of A. karroo are also used as aphrodisiac, for
economic importance in this country [11] while the gum is general body pains, convulsions and dizziness [25]. In
collected and used as a substitute for gum arabic in Zimbabwe Mozambique, root bark infusion of A. karroo is taken orally
[8]. According to van Wyk [12], A. karroo bark and leaf have as remedy for malaria [29]. Gum of A. karroo is used with
commercial potential as remedies for diarrhoea, its exudates Capsicum spp. fruit and vinegar in a plaster dressing for acute
are used as an emollient for conjunctivitis and haemorrhage osteomyelitis [28]. The gum from A. karroo has been used
and also as pharmaceutical aid in solid formulations. The medicinally as emollient and as pharmaceutical aids such as
seeds are traditionally roasted and used as a coffee substitute emulsifiers, stabilisers of suspensions and additives for solid
in southern Africa [13,14]. Gum collected from A. karroo can formulations. In South Africa, the gum of A. karroo has been
be used in the commercial production of sweets and other applied to mouth ulcers and is diluted with water and used as
confectioneries [14]. Over the last three decades, various a mouthwash against oral thrush and sprue [13,30]. Thorns are
attempts have been made to investigate chemical constituents, used to relieve heart pains and for magical purposes [31].
biological activities of A. karroo and its ethnomedicinal uses A. karroo is used in ethnoveterinary medicine for diarrhoea,
in southern Africa. Unfortunately, no comprehensive review of coughs and opthalmia in cattle and dogs [19,32]. Root infusions
this important plant species in southern Africa has been of A. karroo are used in ethnoveterinary medicine as an
published, documenting the species’ biology, traditional uses, antidote to poisoning as a result of cattle and goats eating
phytochemistry and pharmacological properties. Therefore, in Moraea spp. [33]. A. karroo is used to treat cattle which have
this study, the advances in traditional utilization, botany, tulp poisoning, that is poisoning caused by consuming parts of
phytochemistry, pharmacology and safety aspects of A. karroo Homeria spp., a bulbous plant species known to be poisonous
are systematically reviewed. to stock [1,13]. A. karroo provides shade for livestock such as
cattle and goats in southern Africa [8]. The leaves, flowers,
2. Methodology of the review pods and its parasitic mistletoes are excellent fodder for
livestock and game in southern Africa [8]. The wood is an
The literature search was performed from June 2016 to excellent fuel, the bark can be used for tanning, the inner bark
January 2017 using electronic search engines such as Google, makes good cord and the sawn timber can be used for general
Google scholar, publishing sites such as Elsevier, scienceDirect, purposes. A. karroo gum is collected and used as a substitute
BioMed Central (BMC) and PubMed. The databases and liter- for gum arabic is Zimbabwe [8]. Seeds of A. karroo have been
ature sources were chosen based on the topic covered (i.e., used as a substitute for coffee [19].
 Alfred Maroyi/Asian Pacific Journal of Tropical Medicine 2017; 10(4): 351–360 353

Table 1
Ethnomedicinal and other traditional uses of Acacia karroo in southern Africa.

Use Plant part(s) used Country practised Refs

Abscesses Gum applied externally South Africa [28]
Aphrodisiac Root decoction taken orally Zimbabwe [25,26]
Astringent Bark decoction applied externally South Africa [34]
Coagulant Bark decoction applied externally South Africa [28]
Colds Bark decoction taken orally South Africa [19,20]
Colic Crushed roots mixed with food or Lesotho, South Africa [17–20]
root decoction taken orally
Convulsions Root decoction Zimbabwe [25,35]
Diarrhoea Bark, gum and leaf concoctions and South Africa; Zimbabwe [19–24]
infusions taken orally
Dizziness Root infusion taken orally Zimbabwe [25]
Dysentery Bark, gum and leaf concoctions South Africa; Zimbabwe [19,20,22,24]
and infusions
Emetic Bark decoction taken orally South Africa [19,20]
Flu Bark decoction taken orally South Africa [23]
General body pains Body washed with root infusion Zimbabwe [25]
Gonorrhoea Root decoction taken orally Zimbabwe [25,26]
Haemorrhage Bark, gum and leaf concoctions and South Africa [22]
infusions taken orally
Headache Leaf infusion taken orally South Africa [36]
Heart pains Thorn used to relieve pains South Africa [31]
Inflammation of eyes Bark, gum and leaf concoctions South Africa [20]
and infusions taken orally
Magical purposes Thorn used for magical purposes South Africa [31]
Malaria Root bark infusion taken orally Mozambique [29]
Mouth ulcers Gum applied to mouth ulcers South Africa [13,30]
Oral thrush Gum diluted with water and taken orally South Africa [13]
Osteomyelitis Gum mixed with Capsicum spp. South Africa [28]
fruit and vinegar applied in a plaster
Purge symptoms Root decoction taken orally South Africa [19]
of evil and sorcery
Ringworm Bark decoction applied on affected body part South Africa [37]
Sexually transmitted diseases Root decoction taken orally South Africa [27]
Snake repellent Root bark decoction sprinkled to repel snakes South Africa [38]
Stomach ache Bark infusions taken orally South Africa [28]
Syphilis Root decoction taken orally Zimbabwe [25,26]
Urinary schistosomiasis Root decoction taken orally Zimbabwe [24]
Venereal diseases Root decoction taken orally South Africa [20]
Worms Bark or leaf decoction South Africa [39]
Ethnoveterinary medicine
Diarrhoea Bark, leaf and root decoction Namibia, South Africa [40–42]
Ectoparasites Leave root in fowl run Zimbabwe [43]
Fractures Bark decoction applied externally South Africa [40]
Tulp poisoning Bark decoction taken orally South Africa [28]
Wounds and myiasis Leaf decoction applied externally South Africa [44]
Other uses
Coffee substitute South Africa [19]
Cord or rope South Africa [45]
Dye Botswana, South Africa, Zimbabwe [46]
Edible gum South Africa, Zimbabwe [8,47]
Fence Zimbabwe [48]
Firewood Botswana, South Africa, Zimbabwe [31,45,48,49]
Fodder Botswana, South Africa, Zimbabwe [8,46]
Shade Botswana, South Africa, Zimbabwe [8,46]

4. Phytochemical and nutritional constituents of A. high levels of condensed tannins (Table 2), which have been
karroo documented by several other authors such as Mokoboki et al
[52], Dube et al [54], Ngambu et al [55], Gxasheka et al [56] and

The nutritional composition of A. karroo leaves is shown in Brown et al [57]. The inclusion of A. karroo leaves as
Table 2. A. karroo leaves contain high levels of crude protein supplementary feed in the diet of goats and other livestock
and minerals (Table 2) and the crude protein values for the could benefit the smallholder farmers in the communal areas
species are within the optimal range of (120–230) g/kg dry of southern Africa during the critical fodder scarcity.
matter required for body weight gain, maintenance and pro- Phytochemical screenings of various plant parts of A. karroo
duction requirements in growing goats [50,51]. A. karroo leaves demonstrated the presence of flavonoids, phenols, phytosterols,
also have moderate levels of detergent fibres which are proanthocyanidin, tannin and terpenes [54,57,61–64]. Nyila et al [63]
indication of high feeding values [52,53]. A. karroo contains isolated epicatechin 1, b-sitosterol 2 and epigallocatechin 3 from
354 Alfred Maroyi/Asian Pacific Journal of Tropical Medicine 2017; 10(4): 351–360

Table 2 six chemical compounds [64]: 4-methyl-2,4-bis(40 -trimethy-lsily-

Nutritional composition of Acacia karroo leaves. loxyphenyl)pentene 5, cyclotrisiloxane,hexamethyl-trans-decalin
7, 2-methyl-bicyclo[2.2.1]heptan-2-one,4,7,7-trimethyl 8, furan,
Caloric and nutritional Values Refs
composition
2-hexyl 9, cyclohexane,1-methyl-4-(1-methylethenyl) 10 and
decalin,2-methyl 11 (Table 3). The ethanol extract of A. karroo
Acid detergent fibre (ADF) (%) 32.4 [57]
[58]
leaves contained eight chemical compounds [64]: trimethyl[4-
Ash (g/kg DM) 51
Ca 1.73 ± 0.02 [59] (1,1,3,3,-tetramethylbutyl)phenoxy]silane 6, cyclotrisiloxane,
Crude fibre (g/kg DM) 259 [58] hexamethyl-trans-decalin 7, cyclodecene,1-methyl 12, acet-
Crude protein (g/kg DM) 148.9 [52] amide,N-(3-imidazol-1-yl-propyl)-2-methoxy-cyclohexene 13,
Cu (ppm) 10.7 [60]
cyclohexene,1-pentyl 14, 2-methyl-bicyclo[2.2.1]heptan-2-
Dry matter (%) 97.0 [59]
Ether extract (%) 2.4 [57] one,4,7,7-trimethyl 15, methyl-2,4-bis(40 -trimethylsilylox-
Fe (ppm) 175 ± 18 [59] yphenyl)pentene 16 and oxamide,N-[3-(1-imidazolyl)propyl]-N0 -
K (%) 0.970 ± 0.001 [59] methyl 17 (Table 3). The chloroform extract of the roots of A
Mg (%) 0.320 ± 0.001 [59] karroo contained six chemical compounds [64]: decalin,2-methyl
Mn (ppm) 13 ± 3 [59]
10, 2-methyl-bicyclo[2.2.1]heptan-2-one,4,7,7-trimethyl 15,
Na (%) 0.01 ± 0.00 [59]
Neutral detergent 38.0 [57] methyl-2,4-bis(40 -trimethylsilyloxyphenyl)pentene 16, 3-(4,8,12-
fibre (NDF) (%) trimethyltridecyl)furan 18, bicyclo[4.1.0]heptan-3-one,4,7,7-
Organic matter (%) 92.1 [57] trimethyl-,1r-(1a,4a,6a) 19, 1,2-bis(trimethylsilyl)benzene 20,
P (%) 0.13 ± 0.01 [59] trimethyl(4-tert.-butylphenoxy)silane 21 (Table 3). The ethyl ac-
Se (ppm) 0.17 [60]
[59]
etate extract of the roots of A karroo contained six chemical
Tannin (%) 2.220 ± 0.008
Total phenolics (%) 38.0 [57] compounds [64]: cyclotrisiloxane,hexamethyl-trans-decalin 7,
Zn (ppm) 66 ± 2 [59] furan,2-hexyl 9, decalin,2-methyl 11, cyclodecene,1-methyl 12,
bicyclo[4.1.0]heptan-3-one,4,7,7-trimethyl-[1r-(1a,4b,6a) 22 and
spiro[5.5]undecane 23 (Table 3). The ethanol extract of A. karroo
ethyl acetate extracts of A. karroo leaves using silica gel column
roots also contained six chemical compounds [64]:
chromatographic (CC) purification technique (Table 3). A chlo-
cyclotrisiloxane,hexamethyl-trans-decalin 7, cyclodecene,1-
roform crude extract of A. karroo leaves analyzed using the gas
methyl 12, bicyclo[4.1.0]heptan-3-one,4,7,7-trimethyl-,[1r-
chromatography-mass spectrometry (GC-MS) technique [64]
(1a,4a,6a) 19, trimethyl(4-tert.-butylphenoxy)silane 21, bicyclo
yielded three ingredients: cyclohexanone,2-methylene-5-(1-
[4.1.0]heptan-3-one,4,7,7-trimethyl-1r-(1a,4b,6a) 22, 2-
methylethyl) 4, 4-methyl-2,4-bis(40 -trimethylsilyloxyphenyl)pen-
cyclohexen-1-one,2-methyl-5-(1-methylethyl),(S) 24 and 1,2-
tene 5 and trimethyl[4-(1,1, 3,3,-tetramethylbutyl)phenoxy]silane
benzisothiazol-3-amine-tbdms 25 (Table 3).
6 (Table 3). The ethyl acetate extract of A. karroo leaves contained

Table 3
Chemical compounds isolated and characterized from Acacia karroo.

No. Compound Extract Plant part Method of Refs

characterization
1 Epicatechin Ethyl acetate Leaves CC [63]
2 b-sitosterol Ethyl acetate Leaves CC [63]
3 Epigallocatechin Ethyl acetate Leaves CC [63]
4 Cyclohexanone,2-methylene-5-(1-methylethyl) Chloroform Leaves GC-MS [64]
5 4-methyl-2,4-bis(40 -trimethylsilyloxyphenyl)pentene Chloroform, ethyl Leaves GC-MS [64]
acetate, ethanol
6 Trimethyl[4-(1,1,3,3,-tetramethylbutyl)phenoxy]silane Chloroform, ethanol Leaves GC-MS [64]
7 Cyclotrisiloxane,hexamethyl-trans-decalin Ethyl acetate, ethanol Leaves GC-MS [64]
8 2-methyl-(bicyclo[2.2.1]heptan-2-one,4,7,7-trimethyl Ethyl acetate Leaves GC-MS [64]
9 Furan,2-hexyl Ethyl acetate Leaves, roots GC-MS [64]
10 Cyclohexane,1-methyl-4-(1-methylethenyl) Ethyl acetate Leaves GC-MS [64]
11 Trans-decalin,2-methyl Chloroform, ethyl acetate Leaves, roots GC-MS [64]
12 Cyclodecene,1-methyl Ethanol, ethyl acetate Leaves, roots GC-MS [64]
13 Acetamide,N-(3-imidazol-1-ylpropyl)-2-methoxy Ethanol Leaves GC-MS [64]
14 Cyclohexene,1-pentyl Ethanol Leaves GC-MS [64]
15 2-methyl-bicyclo[2.2.1]heptan-2-one,4,7,7-trimethyl Chloroform, ethanol Leaves, roots GC-MS [64]
16 Methyl-2,4-bis(40 -trimethylsilyloxyphenyl)pentene Chloroform, ethanol Leaves, roots GC-MS [64]
17 Oxamide,N-[3-(1-imidazolyl)propyl]-N0 -methyl Ethanol Leaves GC-MS [64]
18 3-(4,8,12-trimethyltridecyl) furan Chloroform Roots GC-MS [64]
19 Bicyclo[4.1.0]heptan-3-one,4,7,7-trimethyl-,[1r-(1a,4a,6a) Chloroform Roots GC-MS [64]
20 1,2-bis(trimethylsilyl)benzene Chloroform Roots GC-MS [64]
21 Trimethyl(4-tert.-butylphenoxy)silane Chloroform, ethanol Roots GC-MS [64]
22 Bicyclo[4.1.0]heptan-3-one,4,7,7-trimethyl-,1r-(1a,4b,6a) Ethyl acetate, ethanol Roots GC-MS [64]
23 Spiro[5.5]undecane Ethyl acetate Roots GC-MS [64]
24 2-cyclohexen-1-one,2-methyl-5-(1-methylethyl),(S) Ethanol Roots GC-MS [64]
25 1,2-benzisothiazol-3-amine tbdms Ethanol Roots GC-MS [64]
 Alfred Maroyi/Asian Pacific Journal of Tropical Medicine 2017; 10(4): 351–360 355

5. Pharmacological activities respectively. Methanol extracts of A. karroo leaves caused the

maximum zone of inhibition against P. vulgaris
A number of pharmacological activities of A. karroo have (20.33 ± 1.53) mm and the lowest against S. typhi
been reported in literature justifying some of its ethnomedicinal (10.33 ± 1.53) mm. An ethyl acetate extract of A. karroo root
uses. These biological activities include antibacterial caused the maximum zone of inhibition against Staphylooccus
[14,20,27,62,64–66], antifungal [14,20,27,65,66], antigonococcal [20], aureus (33.30 ± 1.53) mm and the lowest against E. coli
antihelmintic [24,58,67–69], anti-inflammatory and analgesic (8.67 ± 1.53) mm [64]. Mamba et al [27] evaluated antibacterial
[27,70], antilisterial [63], antimalarial [71], antimycobacterial activities of ethanol extracts of A. karroo against Gardnerella
[62,65], antioxidant [66] and HIV-1 reverse transcriptase [20,27,72]. vaginalis, Neisseria gonorrhoeae and Oligella ureolytica
using the serial broth micro-dilution assay with ciprofloxacin
5.1. Antibacterial as the positive control. Good activity was demonstrated by the
tested microbes with minimal inhibition concentration (MIC)
Mulaudzi et al [20] investigated the antibacterial effects of values of 6.3 mg/mL against G. vaginalis, O. ureolytica (1.6 mg/
aqueous, acetone, dichloromethane, ethanol, methanol and mL) and N. gonorrhoeae (0.8 mg/mL) [27]. Tshikalange et al [66]
petroleum ether bark extracts of A. karroo against Bacillus also evaluated antibacterial activities of ethanol extracts of
subtilis, Escherichia coli, Klebsiella pneumoniae and A. karroo roots against E. coli, Klebsiella oxytoca, K.
Staphylococcus aureus using micro-dilution bioassay with pneumoniae subsp. pneumoniae, N. gonorrhoeae,
neomycin as positive control. The minimal microbicidal con- Staphylococcus aureus using the serial broth micro-dilution
centration (MMC) of the tested bacteria ranged from 0.195 to assay with ciprofloxacin as positive control. Lowest minimal
3.125 mg/mL, with the best activity with MMC value of inhibition concentration (MIC) value were demonstrated against
0.195 mg/mL displayed by aqueous extract against Staphylo- K. oxytoca (0.8 mg/mL), N. gonorrhoeae (0.8 mg/mL) and
coccus aureus [20]. Madureira et al [62] evaluated antibacterial Staphylococcus aureus (0.4 mg/mL) [66]. Chattopadhyay et al
[73] evaluated antibacterial activities of the compound b-
activities of hexane, dichloromethane, ethyl acetate and
methanol extracts of aerial parts of A. karroo against Gram- sitosterol 1 against E. coli, Enterococcus faecalis, P. mirabilis,
positive (Staphylococcus aureus and Enterococcus faecalis) P. aeruginosa, Staphylooccus aureus and Staphylooccus
and Gram-negative bacteria (E. coli, Pseudomonas aeruginosa, saprophyticus using disk diffusion methods with amoxycillin
K. pneumoniae) using the broth microdilution method. The and gentamicin as controls. Weak activity was demonstrated
minimal inhibition concentration (MIC) of the tested bacteria by the tested microbes with minimal inhibition concentration
ranged from 7.5 to >250 mg/mL, with the best activity with MIC (MIC) values ranging from 512 to 1000 mg/mL [73]. These
value of 7.5 mg/mL displayed by methanol extract against antibacterial properties displayed by different extracts of
Staphylococcus aureus [62]. Similarly, Nielsen et al [65] A. karroo somehow confirm the species’ antibacterial potential
evaluated antibacterial activities of leaf and stem methanol and its usefulness in the treatment and management of
extracts of A. karroo against Citrobacter, E. coli, K. bacterial infections such as diarrhoea, dysentery, gonorrhoea
pneumoniae, P. aeruginosa and Staphylococcus aureus using and syphilis, see Table 1.
liquid micro-broth dilution technique with ciprofloxacin as
positive control. The minimal inhibition concentration (MIC) of 5.2. Antifungal
the tested bacteria ranged from 78.12 to 1250.00 mg/mL, with
the lowest MIC value of 78.12 mg/mL displayed by stem extracts Mulaudzi et al [20] evaluated the antifungal effects of
against K. pneumoniae and Staphylococcus aureus [65]. The aqueous, acetone, dichloromethane, ethanol, methanol and
minimal microbicidal concentration (MMC) of the tested petroleum ether bark extracts of A. karroo against Candida
bacteria ranged from 156.25 to >2500.00 mg/mL, with the albicans using micro-dilution bioassay with amphotericin as
lowest MMC value of 156.25 mg/mL displayed by stem positive control. The minimal inhibition concentration (MIC) of
extracts against Staphylococcus aureus (Nielsen et al., 2012). the tested fungus ranged from 3.125 to 6.250 mg/mL, while the
Cock and van Vuuren [14] evaluated antibacterial activities of minimum fungicidal concentration (MFC) values ranged from
methanol and aqueous leaf extracts of A. karroo against 3.125 to >12.500 mg/mL [20]. Nielsen et al [65] also evaluated
Alicaligenes faecalis, Aeromonas hydrophilia, Bacillus cereus, the antifungal activities of leaf and stem methanol extracts of
B. subtilis, Citrobacter freundi, E. coli, K. pneumoniae, A. karroo against C. albicans and Microsporum audouinii
Proteus mirabilis, Proteus vulgaris, P. aeruginosa, using liquid micro-broth dilution technique. The results of the
Pseudomonas fluorescens, Salmonella typhimurium, Serratia minimal inhibitory concentration (MIC) indicated the lowest
marcescens, Shigella sonnei, Staphylococcus aureus and value of 78.12 mg/mL from methanol stem extracts against both
Staphylococcus epidermidis using modified disc diffusion species [65]. The results of the minimal microbicidal
method with chloramphenicol, ampliciilin and nystatin as concentration (MMC) indicated weak activity of 312.50 mg/
positive controls. The minimal inhibition concentration (MIC) mL from methanol stem extracts against both species [65].
values of the tested microbes ranged from 235 mg/mL to Cock and van Vuuren [14] evaluated antifungal activities of
4836 mg/mL, with lowest MIC value of 235 mg/mL methanol and aqueous leaf extracts of A. karroo against
demonstrated by aqueous extracts against B. subtilis [14]. Aspergillus niger, C. albicans and Rhizopus stolonifer using a
Priyanka et al [64] evaluated antibacterial activities of leaf and modified disc diffusion method with chloramphenicol,
root chloroform, ethanol, ethyl acetate and methanol A. karroo ampliciilin and nystatin as positive controls. Antifungal
extracts using the agar well diffusion method against activities were observed in methanol and aqueous leaf extracts
Staphylooccus aureus, E. coli, Salmonella typhi, P. against A. niger with minimal inhibition concentration (MIC)
aeruginosa, K. pneumoniae, P. vulgaris and B. subtilis with values of 486 mg/mL and 325 mg/mL, respectively [14].
ampicillin and distilled water as positive and negative controls, Recently, Mamba et al [27] evaluated antifungal activities of
356 Alfred Maroyi/Asian Pacific Journal of Tropical Medicine 2017; 10(4): 351–360

ethanol extracts of A. karroo against C. albicans using the serial et al [69] found that the cattle that received the A. karroo leaf
broth micro-dilution assay. The extract demonstrated good ac- meal had lower H. contortus and O. colombianum worm
tivity with minimal inhibition concentration (MIC) value of burdens than those that received the control diet. Marume et al
0.8 mg/mL [27]. In a separate study, Tshikalange et al [66] [58] evaluated antihelmintic effects of A. karroo leaf extracts in
evaluated antifungal activities of ethanol extracts of A. karroo four month old Xhosa lop-eared goats exposed to a single dose
roots against C. albicans. The extract demonstrated some of 6000 freshly cultured L3 H. contortus larvae. Marume et al [58]
activity with minimal inhibition concentration (MIC) value of observed reduction in faecal larval counts and H. contortus worm
1.6 mg/mL against the fungus [66]. counts in goats that consumed A. karroo leaves (182 g/d). Based
on these evaluation reports, A. karroo presents an inexpensive,
5.3. Antigonococcal risk-free and eco-friendly approach to controlling worm popula-
tion in livestock in rural areas and other marginalized commu-
Mulaudzi et al [20] evaluated the antigonococcal activities of nities. According to Brown et al [57], the use of A. karroo for
aqueous, acetone, dichloromethane, ethanol, methanol and helminthic control can be used as a feasible alternative to
petroleum ether bark extracts of A. karroo against N. commercially manufactured antihelmintic or as part of an
gonorrhoeae through determination of clear zones of integrated system to reduce future occurrences of antihelmintic
inhibition with ciprofloxacin and dimethylsulfoxide (DMSO) resistance to commercial medicines.
as positive and negative controls respectively. A. karroo
showed moderate activity with dichloromethane, ethanol and 5.5. Anti-inflammatory and analgesic
petroleum ether extracts with %inhibition ranging from
44.0 ± 0.0 to 55.0 ± 2.0 [20]. The good activity observed from Adedapo et al [70] evaluated anti-inflammatory activities of
the plant extracts tested in this study could lead to the the aqueous extract of the stem bark of A. karroo using the
isolation of lead antigonococcal compounds. carrageenan-induced and histamine-induced rat paw oedema
models and analgesic activity was evaluated using acetic acid-
5.4. Antihelmintic induced writhing response in mice. The extract at 100 and
200 mg/kg reduced significantly the formation of oedema
Sparg et al [67] evaluated the antihelmintic effects of induced by carrageenan and histamine [70]. In the acetic acid-
A. karroo leaf extracts against schistosomules of the species induced writhing model, the extract showed a good analgesic
Schistosoma haematobium. A. karroo extracts at 50 mg/mL effect characterized by a significant reduction in the number of
killed 33% of schistosomula worms after 1 h, 66.7% of the writhes with two doses (100 and 200 mg/kg) used when
worms were killed at 25 mg/mL and A. karroo extracts were compared to the untreated control group [70]. In the tail
100% lethal at 12.5 mg/mL [67]. Mølgaard et al [24] evaluated immersion test, the extract at the doses used (100 and 200 mg/
the antihelmintic effects of A. karroo leaf and root extracts kg) increased reaction time to pain after 30 min of oral
against schistosomules of the trematode Schistosoma mansoni administration of the extract. Mamba et al [27] evaluated anti-
and cysticercoids of the cestode Hymenolepis diminuta. The inflammatory activities of ethanol extracts of A. karroo by
extracts killed the newly excysted cysticercoids within an determining the inhibitory effect of the extracts on the activities
hour, when incubated in a culture medium. The lethal of the pro-inflammatory enzyme, lipoxygenase and inducible
concentrations of A. karroo extracts varied from 0.8 to nitric oxide synthase with quercetin and dimethyl sulphoxide
17.0 mg/mL after 24 h [24]. The best results against H. (DMSO) as positive and negative controls respectively. A.
diminuta were obtained with leaf extracts with lethal karroo showed good 15-LOX inhibition activity with IC50 value
concentrations of 3.1 mg/mL and 0.8 mg/mL after 1 h and of 62.24 mg/mL, which is comparable to the IC50 value of the
24 h, respectively. A. karroo extracts showed some activity positive control quercetin which was 48.86 mg/mL [27].
against S. mansoni with lethal concentrations varying from Chattopadhyay et al [73] evaluated anti-inflammatory activ-
0.25 to 0.30 mg/mL [24]. A. karroo leaf extract was also tested ities of the methanol extracts of compound b-sitosterol 1 using
against schistosomules showing weak activity with lethal carrageenan-induced rat paw oedema (acute model), dextran-
concentrations of 103.0 mg/mL [24]. These pharmacological induced rat paw oedema (sub-acute model) and cotton pellet-
evaluations are of importance in the traditional use of induced granuloma (chronic model) with indomethacin as con-
A. karroo as an antihelmintic [39] and as herbal medicine trol. The anti-inflammatory activity of b-sitosterol 1 demon-
against urinary schistosomiasis [24] and future research strated maximum inhibition of 64.39% at 25 mg/kg dose in
focusing on control and management of schistosomiasis in carrageenan-induced rat paw oedema against 67.47% inhibition
sub-Saharan Africa. demonstrated by the standard indomethacin after 3 h of drug
Kahiya et al [68] evaluated the antihelmintic effects of treatment. In the dextran-induced rat paw oedema model, b-
A. karroo leaves (200 g/d) on Boer goats orally infected with a sitosterol 1 showed inhibition of 60.48% at 25 mg/kg nearly
single dose of Haemonchus contortus third stage larvae. Kahiya equal to the inhibition of 60.73% produced by indomethacin.
et al [68] observed a 34% decrease in the faecal egg counts in The results of the cotton-pellet granuloma model of inflamma-
A. karroo fed animals relative to the control group. Similarly, tion that b-sitosterol 1 significantly inhibited the granuloma
Xhomfulana et al [69] evaluated antihelmintic effects of weight in a dose dependent manner with a maximum inhibition
A. karroo leaf meal on H. contortus and Oesophagostomum of 51.54% at 25 mg/kg compared to 54.07% for indomethacin
colombianum in cattle. Faecal samples were collected from the [73]. These results gave a scientific basis to the traditional uses of
cattle recta every fortnight and examined for nematode egg A. karroo mainly for wound poultices, eye treatments and cold
types using the modified McMaster technique. Xhomfulana remedies.
 Alfred Maroyi/Asian Pacific Journal of Tropical Medicine 2017; 10(4): 351–360 357

5.6. Antilisterial activity documented antioxidant activities A. karroo root extracts are
probably due to flavonoids and phenols that have been isolated
Nyila et al [63] evaluated antilisterial activity of ethyl acetate from leaves [54,57,61–64]. Flavonoids and phenolic compounds
and chloroform extracts of A. karroo against Listeria found in plants are known to have antioxidant properties [75].
monocytogenes using the disc diffusion method with
erythromycin as positive control. The ethyl acetate extract of 5.10. HIV-1 reverse transcriptase
A. karroo showed good antilisterial activity, exhibiting both
minimum inhibitory concentration (MIC) and minimum Mulaudzi et al [20] evaluated anti-HIV activities of aqueous and
bactericidal concentration (MBC) of 3.1 mg/mL, while MIC methanol bark extracts of A. karroo using a non-radioactive HIV-1
and MCB for chloroform extract were 6.25 mg/mL [63]. The RT colorimetric ELISA kit. The aqueous and methanol extracts of
same authors also evaluated the antilisterial activities of ethyl A. karroo bark showed good HIV-1 reverse transcriptase (RT)
acetate extracts of three compounds namely epicatechin 1, b- inhibition percentage (70%) at 1 mg/mL based on COX-assay,
sitosterol 2 and epigallocatechin 3 isolated from A. karroo with all tested extracts exhibiting dose dependent IC50 values of
against L. monocytogenes using the disc diffusion method with (0.03 ± 0.00) and (0.10 ± 0.01) mg/mL, respectively [20]. Moll et al
erythromycin as positive control. The three compounds [72] evaluated anti-HIV activities of 50% methanol:dichloro-
demonstrated good antilisterial activities with MIC and MCB methane (1:1) leaf and twig extracts of A. karroo using a reverse
values ranging from 0.031 to 0.500 mg/mL [63]. transcriptase test kit. A. karroo demonstrated some inhibitory ac-
tivity of reverse transcriptase [72]. Recently, Mamba et al [27]
5.7. Antimalarial evaluated anti-HIV activities of ethanol extracts of A. karroo
against recombinant HIV-1 enzyme using non-radioactive HIV-
Ramalhete et al [71] evaluated antimalarial activity of n- RT colorimetric assay with doxorubicin as positive control. A.
hexane, dichloromethane, ethyl acetate and methanol extracts karroo demonstrated moderate inhibition of HIV-1 reverse tran-
of aerial parts of A. karroo against Plasmodium falciparum. A. scriptase activity with 66.8% inhibition compared to 96.5%
karroo showed moderate to no significant activity with IC50 inhibitory activity demonstrated by doxorubicin, the positive
values ranging from (60.00 ± 12.30) mg/mL to > 100 mg/mL control. Therefore, the good inhibitory activity on HIV-1 reverse
[71]. However, it is important to note that A. karroo is transcriptase demonstrated by A. karroo extracts may imply that
frequently used to treat fever or malaria in Mozambique [29] the species could be a good source of potent compounds for
and therefore, an explanation for their lack of significant therapeutic strategy against HIV-1 reverse transcriptase.
in vitro antimalarial inactivity could be that these plants may
act as antipyretics or may enhance the immune system, rather 5.11. Cytotoxicity and toxicity
than having direct antiparasitic activity [74].
The aqueous extract from the shoot of A. karroo was eval-
5.8. Antimycobacterial uated for its acute toxicity by the oral route in mice and for the
sub-acute effect on haematological, biochemical and histological
Madureira et al [62] evaluated antimycobacterial activities of parameters in Wistar rats [76]. In the acute toxicity test, A. karroo
hexane, dichloromethane, ethyl acetate and methanol extracts of extract caused death in animals that received 1600 and 3200 mg/
aerial parts of A. karroo against Mycobacterium smegmatis kg doses. Oral treatments in rats with this extract at 800 mg/kg
using the broth microdilution method. The minimal inhibition did not cause any significant change in the red blood cell count
concentration (MIC) of the tested bacterium ranged from 31.0 (RBC), packed cell volume (PCV), haemoglobin concentration
to >250.0 mg/mL, with the best activity with MIC value of (HB), mean corpuscular volume (MCV), mean corpuscular
31.0 mg/mL displayed by n-hexane extract [62]. Similarly, haemoglobin concentration (MCHC), mean corpuscular
Nielsen et al [65] evaluated antimycobacterial activities of the haemoglobin (MCH), white blood cells and its differentials. It,
stem methanol extract of A. karroo against M. smegmatis and however, caused a significance decrease in the levels of
Mycobacterium tuberculosis using the radiometric respiratory platelets [76]. In the biochemical parameters, the extract caused
techniques with dimethylsulfoxide (DMSO) as control. Both a significant decrease in the levels of total protein, albumin,
M. smegmatis and M. tuberculosis demonstrated weak activity globulin, aspartate amino transferase (AST), alanine amino
with minimal inhibition concentration (MIC) values of 1250 transferase (ALT), total and unconjugated bilirubin. Adedapo
and 2500 mg/mL, respectively [65]. Therefore, these et al [76] noted changes in the body weights of the mice but
preliminary evaluations done by Madureira et al [62] and no significant changes were observed in the levels of some
Nielsen et al [65] provide baseline data for future research on electrolytes (sodium, potassium and chloride). Lung with
the species as a possible source of traditional medicine for multiple abscess, kidney and liver with mild congestion were
treatment of tuberculosis and other respiratory ailments. also observed histopathologically [76]. Cock and van Vuuren
[14] evaluated toxicity of aqueous and methanol leaf extracts of

5.9. Antioxidant A. karroo using a modified Artemia franciscana nauplii

lethality assay. A. karroo leaf water and methanolic ectracts
Tshikalange et al [66] evaluated antioxidant activities of induced mortalities in the Artemia nauplii below 20%
ethanol extracts of A. karroo roots by assessing the free radical following 24 h and 48 h of exposure, indicating that the
scavenging activity using DPPH (2, 2-diphenyly-1- extracts are of low toxicity.
picrylhydrazyl) with ascorbic acid (vitamin C) as a positive Nyila et al [63] evaluated the cytotoxicity of ethyl acetate and
control. The IC50 of the extract was 0.83 mg/mL, while vitamin C chloroform extracts of A. karroo using the XTT method
(positive control) had an IC50 value of 1.44 mg/mL [66]. The using the cell proliferation kit II (Boehringer-Mannheim) with
358 Alfred Maroyi/Asian Pacific Journal of Tropical Medicine 2017; 10(4): 351–360

zearalenone as positive control. Epicatechin 2 was the least toxic [2] Palmer E, Pitman P. Trees for southern Africa covering all known
compound with IC50 value of >200.0 mg/mL, while b-sitosterol indigenous species in Republic of South Africa, South West Africa,
1 and epigallocatechin 3 were found to be 63.82 and 28.91 mg/ Botswana, Lesotho and Swaziland. Cape Town: A.A. Balkema;
1972, p. 792.
mL, respectively [63]. Tshikalange et al [66] evaluated
[3] Dyer C. New names for the African Acacia species in Vachellia
cytotoxicity activities of ethanol extracts of A. karroo roots on and Senegalia. South For A J For Sci 2014; 76(1): 4.
Vero African monkey cells lines with 2, 3-bis[2-methoxy-4- [4] Maslin BR. Generic and subgeneric names in Acacia following
nitro-5-sulfophenyl]-2H-tetrazolium-5-carboxanilide (XTT) re- retypification of the genus. Muelleria 2008; 26(1): 7-9.
agents with actinomycin D as positive control. The extracts [5] Moore G. Action on the proposal to conserve the name Acacia at
showed toxicity with 50% viability of cells (EC50) at concen- the nomenclature section of the XVII international botanical
trations value of 115 mg/mL against actinomycin D which was congress in vienna: did a yes have it? Linnean 2008; 24(1): 16-120.
[6] Taylor CL, Barker NP. Species limits in Vachellia (Acacia) karroo
used as positive control exhibited an IC50 value of 0.009 32 mg/ (mimosoideae: Leguminoseae): evidence from automated ISSR
mL [66]. These preliminary cytotoxicity and toxicity evaluations DNA ‘fingerprinting’. S Afr J Bot 2012; 83(1): 36-43.
carried out so far [14,63,66,76] study concluded that caution must [7] Idamokoro EM, Masika PJ, Muchenje V. Vachellia karroo leaf
be exercised in the use of the plant for medicinal purposes. meal: a promising non-convential feed resource for improving goat
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6. Conclusions Range Forage Sci 2016; 33(3): 141-153.
[8] Barnes RD, Filer DL, Milton SJ. Acacia karroo: monograph and
annotated bibliography. Oxford: Oxford Forestry Institute, Uni-
A. karroo has been used in southern Africa as herbal medi- versity of Oxford; 1996, p. 97. Tropical Forestry Paper No. 32.
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limited. More research is required and future research should development and innovation. Queensland: Department of Employ-
focus on more comprehensive chemical characterization of both ment, Economic Development and Innovation, Queensland Gov-
ernment; 2010, p. 15.
crude and pure extracts, evaluate potential for commercialization
[10] Ross JH. A conspectus of the African Acacia species. Pretoria:
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The author declares that he has no conflict of interest. plants used for the treatment of diarrhoea in the Eastern Cape,
South Africa. Pak J Biol Sci 2008; 11(15): 1961-1963.
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