FROM TOPS TO BOTTOM: USING DNA BARCODING TO COMBAT THE THE Formatted: Line spacing: Double

ILLEGAL HARVEST

OF THREATENED PLANT SPECIES IN SOUTH AFRICA AS RECORDED

IN THE NATIONAL ENVIRONMENTAL MANAGEMENT: BIODIVERSITY

ACT, 10 OF 2004 TBA TOPS LIST 2015. Commented [S1]: We agreed that you remove FROM
TOPS TO BOTTOM: and also write NEM:BA in full
by Formatted: Line spacing: Double

Mosshadi Mashangoane

are

Dissertation submitted in fulfilment of the requirements for the degree

HONOURS BACHELOR OF SCIENCE

in

Botany

in the

Faculty of Science

at the

University of Johannesburg

Supervisor: Prof Prof. Michelle van der Bank

Co-supervisor: Ms Lerato Hoveka

November 2017
 DECLARATION

I declare that this dissertation hereby submitted to the University of Johannesburg for the

degree BACHELOR OF SCIENCE HONOURS (Botany), is my own work and has not

been previously submitted by me for a degree at another institution.

TABLE OF CONTENTS

Formatted Table
INDEX TO TABLES
LIST OF ABBREVIATIONS Formatted: Left

BOLD - Barcode of Life Database Formatted: Font: Not Bold, Font color: Dark Blue
CITES - Convention on International Trade of Endangered Species of Fauna and Flora Formatted: Font: Not Bold, Font color: Dark Blue
IUCN - International Union for Conservation of Nature
MatK - Maturase K Commented [S2]: Rather use the table to have smooth
alignment, and then select the table, choose no border
option

NEM: BA - National Environmental Management: Biodiversity Act Formatted: Font: Not Bold
Formatted: Font: Not Bold
NFTP - Non- Timber Forest Product
Formatted: Font: Not Bold

rbcLa - Ribulose-1,5-biphosphate Carboxylase/ Oxygenase Formatted: Font: Not Bold

TOPS - Threatened or Protected Species

WWF - World Wildlife Fund

Formatted: Font: Not Bold

Formatted: Left
ACKNOWLEDMENTS Formatted: Left

I would like to thank SANBI for sponsoring my honours degree studies at the university Formatted: Font: Not Bold
Formatted: Font: Not Bold, Font color: Dark Blue
of Johannesburg. I also would like to thank my supervisor Prof Michelle van der Bank for
Formatted: Justified

her supervision and allowing me to be part of a wonderful team at the African Centre for Formatted: Font: Not Bold
Formatted: Font: Not Bold
DNA Barcoding. I would also like to thank Ms Lerato Hoveka for being my co-supervisor. Formatted: Font: Not Bold
Formatted: Font: Not Bold
Lastly, I thank all the lab technicians for assisting me with my lab work. Without all the
Formatted: Font: Not Bold
above-mentioned people, this paper would not exist. Formatted: Font: Not Bold

Formatted: Left

ABSTRACT
Background: Overexploitation of natural resources is a major threat to biodiversity as it
may lead to the extinction of rare and vulnerable species. According to the National
Environmental Management: Biodiversity Act, 2004 (Act No 10 of 2004)
[Source/Reference], National Environmental Biodiversity Act (NEM: BA), act 10 of 2004
any person who is in possession of a species that is listed as protected or threatened or
owns any product derived from these species must have a valid permit to do so
[Source/Reference]. Countries with unique floras such as South Africa face major
challenges in protecting indigenous plants from specialist collectors while having to
manage other threats such as habitat destruction, biological invasions and climate
change [Source/Reference]. DNA barcoding can be used as a tool in aiding plant
conservation. In this study, core DNA barcoding regions (rbcLa and matK) were used to
assist in identifying the threatened plant species of South Africa [Source/Reference].
Results: Based on genetic divergence, PCR amplification efficiency and the BLAST
algorithm [Source/Reference], the core DNA barcodes proved to be efficient
[Source/Reference]. Hence a DNA barcoding library was created on BOLD (Barcode of
Formatted: Left
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later and it will be different from the one you submitted to
the conference.
Formatted: Line spacing: Double
Commented [S4]: Please reference the source or the link
where the NEM: BA (Act No 10 of 2004) can be found
Formatted: Highlight
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be reference from someone who is a subject matter
authority
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be reference from someone who is a subject matter
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be reference from someone who is a subject matter
authority
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be reference from someone who is a subject matter
authority
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be reference from someone who is a subject matter
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be reference from someone who is a subject matter
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Life Database) to assist in future identification of unknown or taxonomically doubtful Commented [S16]: This is a bold statement and needs to
be reference from someone who is a subject matter
authority
species [Source/Reference],. Significance: Plant diversity is not only crucial in
Commented [S17]: This is a bold statement and needs to
maintaining healthy and sustainable ecosystems [Source/Reference], but also holds be reference from someone who is a subject matter
authority

socio-economic value [Source/Reference]. In order to implement strategies to protect Commented [S18]: This is a bold statement and needs to
be reference from someone who is a subject matter
authority
threatened species, identification of these species is a critical first step
Commented [S19]: This is a bold statement and needs to
be reference from someone who is a subject matter
[Source/Reference],. authority
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Chapter 1HAPTER 1 Formatted: Left, Line spacing: Double
Formatted: Font: Bold
1. General Introduction and objectives Formatted: Line spacing: Double
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GENERAL INTRODUCTION AND OBJECTIVES
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1.1 Overexploitation of natural resources as a threat to global biodiversity Formatted: Left, Line spacing: Multiple 1.08 li
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Many species are becoming extinct and biodiversity is wiped out at a fast rate subheadings but please confirm this with Lerato
Formatted: Font: (Default) Arial, 12 pt
[Source/Reference]. Biodiversity is defined as the variety of life on earth which include all
Formatted: Font: (Default) Arial, 12 pt
the species as well as their habitat [Source/Reference]. Furthermore, it can be measured Commented [S25]: This is a bold statement and needs to
be reference from someone who is a subject matter
authority
in terms of generic diversity and the identity structure of each species (Swinggland,2002).
Commented [S26]: This is a bold statement and needs to
be reference from someone who is a subject matter
Overexploitation of natural resources is a major threat to biodiversity as it may lead to the authority
Commented [S27]: Very excellent way of introducing the...
extinction of rare and vulnerable species (Schippmann et al. 2002; Courchamp 2006). Commented [S28]: This is a bold statement and needs to...

According Seshrest and Brooks (2002) extinction of species is the most obvious Commented [S29]: This is a bold statement and needs to...
Commented [S30]: This is a bold statement. Its great that...
biodiversity loss. According to the (WWF, 2017), environmental management and
biodiversity field specialists have reported that at least 10 000 species go extinct every
year. Moreover, the rapid loss of species was estimated to be 1000 to 10000 higher than
natural extinction rate (WWF, 2017).
Ding (2011) avers that as human population increase, overexploitation of natural
resources increases due to over-consumption. Reichard and White (2001) reported that
since the beginning of time (from 8000 BC) Non-timber forest products (NFTPs) have
been exploited by indigenous communities for subsistence and trade. However, in the
past two decades, the commercialization of natural resources including medicinal plants
has increased (Ticktin,2004). The aforesaid increase is attributed to the fact that rural
communities in developing countries depend greatly on NFTPs for their livelihood (Twine
et al. 2003).
Additionally, factors such as the growing number of black people living in the cities have
also contributed greatly to the over-harvest. Most indigent black peopleof them consult
traditional healers since western medicines are too expensive and some peoples beliefs
do not agree with Western medicinese (Williamson et al., 2000). The trade in NFTPs
ranks among the top 5 most profitable yet unlawful trade alongside drug smuggling,
weapons, counterfeit goods as well as human trafficking (Lawson &Vines, 2014). The wild
life trade is permitted for some species, provided that the person has a permit from
authorities [Source/Reference]. However, the problem arises when the trade is done
unsustainably making other species to become vulnerable and putting them in danger of
being extinct [Source/Reference]. The unsustainable trade of species does not only affect
the species being exploited, it also affects the livelihood of the people particularly the poor
Commented [S31]: I have shortened he sentence.
Commented [S32]: Please introduce NFTP by defining
itsimilarly to the way you have introduced overexploration
Commented [S33]: These should be two different
sentences.
Commented [S34]: Please make sure that your intext
references is consistent and correct e.g. (Williamson et al.,
2000) vs (Twine et al. 2003).
Commented [S35]: Please break this into two sentenced.
As a rule of thumb, sentence should have at most 14 words.
Commented [S36]: This sentence is too long
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be reference from someone who is a subject matter
authority
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[Source/Reference]. Moreover, the trade affect other species and the illegal harvesting of
timber plants affects the forest. The overexploitation of this wildlife plants results in loss
of biodiversity (WWF,2017).
Examples of illegally harvested plants
The plant species that are poached are usually sold as ornamentals, crafts and
medicines (Ticktin,2004). Hydrastis canadensis and Cimicifuga racemesa are two
examples of the traditional medicines which are harvested for different purpose.
An example of the aforesaid among others include treating of ailment such as
hemorrhoids and pain respectively. Roberson (2008) reported that in a year more than
60 million of the Hydratis canadensis is collected without being replaced as a consequent
the population of this species has been reduced. It is currently considered to be a plant
of conservation concern by CITES and it is recorded in appendix II of CITES regulations
(Davis and Mc Coy,2000).
Approximately 100 percent of Cimicifuga racemesa comes from wild harvesting
[Source/Reference]. In South Africa, the Encepharlartos population is declining at a fast
rate [Source/Reference]. It has been reported by Williamson et al., (2016) that almost
two-thirds of South Africas Encephalartos species is illegally traded at the traditional
black markets. This is due to the belief that Encephalartos species are useful in chasing
evil spirits away [Source/Reference]. Brackenridgea zanguebarica which is also known
as the magic tree is used by Venda people due to their belief that it possesses magical
powers and healing properties. These species have declined over years and has been
classified as vulnerable according to the IUCN red list categories South Africa
Commented [S41]: This is a bold statement and needs to
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Commented [S43]:
Commented [S44]: Never start a sentence with e.g. (for
an example)
Commented [S45]: Never start a sentence with For
Commented [S46]: Never start a sentence with For"
Commented [S47]: Introduce CITE properly by defining it
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be reference from someone who is a subject matter
authority
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be reference from someone who is a subject matter
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Commented [S51]: Define this scientific tern
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be reference from someone who is a subject matter
authority
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be reference from someone who is a subject matter
authority
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be reference from someone who is a subject matter
authority
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be reference from someone who is a subject matter
authority
Commented [S56]: This is an excellent way of explaining
or describing briefly the scientific term. Please do the same
for the Encephalartos
(Tshisikwawe and Rooyen,2012). The overexploitation of these plants results in

biodiversity loss (WWF,2017)

A B

C D

Formatted: Tab stops: 5.29", Left

Figure 1: Species which are illegally harvested: (A) Hydrastis canadensis (Credit: Lynch,

K.A. and Jones, E.T.). (B) Cimicifuga racemesa (Credit: J. Coddington). (C) Confiscated

Encerphalartos spp. (Credit: Eastern Cape Department of Economic Development and Formatted: Font: Italic

Environmental Affairs. (D) Brackenridgea zanguebarica (Credit: Tshisikwawe, M.P and

Van Rooyen, M.W.).

The significance of biodiversity

Biodiversity is enormously significant to human beings because they depend on other

species to an extent that the loss such species could mean loss of health and wealth and

thus threaten the survival of humans (Barker, 2011). Biodiversity is also very significant

for environmental management and sustainability, as an essential major factor in the

maintenance of the ecosystem [Source/Reference]. It also offers natural capital which is Commented [S57]: I have paraphrased this sentence,
please see if you agree or Lerato agrees with my suggestion
critical in providing ecosystem services [Source/Reference]. Most of the critically

endangered species hotspots are found in the developing countries [Source/Reference]. Commented [S58]: Adding this complete the meaning of
this sentence
Increase loss of endangered species could destabilize the economy of the developing

countries by negatively affecting tourism, which is one of their main sources of national

income [Source/Reference]. The loss of biodiversity worsens poverty similarly poverty is Commented [S59]: I have paraphrased this sentence,
please see if you agree or Lerato agrees with my suggestion
also a major threat of biodiversity (Sligenburg et al., 2009). Commented [S60]: I have paraphrased this sentence,
please see if you agree or Lerato agrees with my suggestion
The diversity of plants is also significant to humans because plants are used for different

things such as medicines, food. Plant ,diversity help to regulate climate change, provide
ecosystem services and most importantly, it is highly valued for its aesthetic features
Commented [S61]: Maybe you should introduce a
sentence which explains/link the plant as part of
endangered species, and please reference the source of
such a bold statement.

(Mutia,2009Sligenburg et al., 2009). Maybe you need to explain biodiversity of plants relying on
relevant research papers/articles and authorities

South Africas biodiversity and its important

According to Germishuizen and Meyer (2003), South Africa is rich in the diversity of

plants as it has got the most abundant flora in the entire world. SANBI (2017) figures

show that South Africa comprises approximately 21000 species which constitute around

8% of the worlds vascular plants. Staden and Raimono (2017) added that 2 576 species

are threatenedd. Furthermore, South Africa also has three biodiversity hotspots

(SANBI,2017). These are the areas which have high amount of species consisting of both

animals and plants which are endemic to the area. The species in these regions are

threatened, endangered, rare and/ or combination of these attributes (Reid, 1998).

According to Taylor and Crous (2001) South Africa has a unique biodiversity hotspot

namely Cape Floristic Region. The Cape Floristic Region is one of the six plant kingdoms

located entirely within the borders of South Africa. Although Cape Floral kingdom is the

smallest of all 6 kingdoms, it has high diversity of species richness and endemism.

However, it has the highest number of threatened or endangered species in the world

(????? Portfolio Review,2004). Furthermore,South Africa is known as the third most Commented [S62]: Theres a missing operative word, e.g
Biodiversity Portfolio Review, 2004)
biodiverse country after Brazil and Indonesia [Source] and it is the only country with more Commented [S63]: Country not countries
than one biodiversity hotspot [Source]. Hence its conservation is very important not only

for South Africa but for the entire world (EThekwini Municipality, 2009).

Other threats to Biodiversity

There are other factors that contributes to the biodiversity besides overexploitation of

plants [Source]. The threats are brought by the anthropogenic factor and some are

brought by natural factors [Source]. [Please write a sentence that introduces or link the

Invasive alien species, Habitat loss and Climate change] as part of biodiversity and

overexploitation.

Invasive alien species

Illegal trading has a potential to cause the increase in the number of species that get Commented [S64]: Introduce and define Illegal trade in
context of biodiversity and endangered species
introduced into a new region which is not their natural habitat. These kind of species are

known as Invasive alien species and they are defined as non-native species introduced

to a certain region by human activities either intentional or unintentionally.

Furthermore, these species pose a negative impact on the environment as it can quickly

outcompete indigenous species (Weber, 2003, Mutia ,2009). Chamier (2012) reported

that invasive alien species are the driver to the loss of biodiversity which have a potential

to cause harm to the terrestrial as well as aquatic ecosystem. Alien invasive species are
known to be the second most common threat which has driven other species to extinction

(IUCN, 2017).

Habitat loss

Habitat loss is one of the main threat to biodiversity caused by human activities [Source]. Commented [S65]: The first sentence must introduce
Habitat loss by defining its meaning
Population growth leads to habitat loss because more people increase pressure on Commented [S66]: Delete which was

natural resources [Source]. Moreover, the population is growing faster thaIt n natural

resources can replace itself [Source]. Habitat loss involves the conversion of land for

other use, which include - (a) production of agricultural crops and (b) creating space for

urban development and industry (Tschakert et al., 2017). Commented [S67]: This should be one complete
sentence
Formatted: Font: Italic
Habitat loss does not only destroysdestroy the individual animals and plant species. Habit

loss also threatens long term survival of the species that were not directly targeted by

fragmenting the habitats [Source]. Consequent to habitat fragmentation, animals become

vulnerable because of environmental fluctuations and diseases [Source]. The animals

find it hard to reach other individuals for mating purposes (Sechrest and Brooks,2002)..

Climate change

Miller and Spoolman (2015) reported, climate has changed drastically over the past 3.5 Commented [S68]: Never start a sentence with AS

billion years. It has been influenced by factors such as volcanic eruptions, shifts in tectonic

plates and changes in the solar input [Source]. Recent studies [Source] have shown that

human activities are the contributors of climate change due to burning of fossil fuels

[Source] and other activities which release greenhouses gases [Source]. These

greenhouse gases have altered the atmospheric variability as they absorb the heat from

sunlight and preventing it from escaping it back into the space (IPCC, 2007).
The change in climate patterns have an impact on the terrestrial biodiversity [Source].

Longer term variability in climate as well as ecosystem health influence the shifts in

distributions of plants, pathogens, animals and even settlement [Source]. Moreover, the

weather events such as flooding and droughts directly impact the ecosystem health,

productivity availability of goods and services which is available for human use (World

Health Organization, 2017).

Management

There are different international organizations were established specifically for the

protection wildlife and management of global trade of the species. The organizations

include CITES, WWF and IUCN to which South Africa is party to (Pandy et al.,2014).

South Africa has drafted specific goals for protecting wild life and management of trading

endangered species globally that are targeted to be achieved by 2020 [Source]. These

goals are classified as target 2, which has a national red list that contains the conservation

status of each plant [Source]. South Africa achieved and reached its target for the target

2 goals, in 2009 [Source]. Conservation status is important as it guides a wide range of Commented [S69]: See how I have broken this into three
different sentences to make it brief and on point. Please
follow this example through the document.
conservation intervention such as land-use decision making and protected area

expansion plan (Raimondo, 2015.;., Staden and Raimondo, 2017).

Making the community of the aware of the biodiversity crisis is a very crucial

thereforecrucial therefore, South Africa has been involved in projects which offers

education programmes about conservation. For example, Silverglen medicinal plant

nursey offers free education programmes about conservation. Furthermore, the

traditional healers and muthi gatherers are taught techniques to cultivate and identify

plants using morphological characters (Ethekweni municipality,2009).

Since key to sustainable environmental management is compliance to the laws and

regulations, South Africa has laws which supports biodiversity conservation (Department

of Environmental Affairs and Tourism, 1998). NEM: BA 10 of 2004 is a South African law

that monitor the species which have been illegally harvested species and ensure that

resources are used in a sustainable manner (Strydom and King,2009) by restricting any

activities which involves threatened or protected species, or have any product derived

from those species (NEMBA, 2015). In addition, South Africa has Protected areas which

are defined under National Environmental Protected Areas Act 57 of 2003. The act

includes special nature reserves, national parks, nature reserves and protected

environment declared under the Act (Paterson,2009). The Act declares that certain areas

must be protected for the protection of ecologically viable areas and to protect the species

that are conservation concern. A permit is required to be in possession of any species

within those areas (Strydom and King,2009).

DNA barcoding

[Define the DNA barcoding]

DNA barcoding is a rapid, accurate and a reliable technique which is developed to help

identify species. It uses a short fragment of gene from chloroplast region in the case of Commented [S70]: This should go to the first sentence,
where I said define DNA barcoding
plants and mitochondrial region in animals. It can identify species that are not wholedoes

not come intact particularly those that , and in such cases, it would be difficult to identify

the species by using morphological characteristics (Panday et al.,2014). In DNA

barcoding, only a small fragment of a specific gene gets sequenced. DNA barcoding only

requires tiny number of samples. The advantages of this technique are thatIt it is

applicable in many ways., Ffor instance, it can be used to detect food fraud, monitor the Commented [S71]: Never start a sentence with for
instance
quality of water and others (Dudu et al., 2016) and. tTaxonomic expert is not that much

necessary to identify the species once a solid database is used (Harvig et al.,2015). DNA

barcoding proved to be successful in identifying the Encerphalatos species that are sold

illegally at the muthi market (Williamson et al., 2016). Commented [S72]: This should be the first paragraph and
should go to the section where I said define DNA barcoding
Formatted: Font: Italic, Font color: Accent 2
[Define in full (NEMBA,2015)]. Although South Africa has good regulations regarding

conservation of species like National Environmental Biodiversity Act (NEMBA) which

restrict any activities involving protected and threatened species (NEMBA,2015). Without
being able to identify species, the regulation would not be effective thus a technique like
DNA barcoding is necessary [Source]. In this study, the species which are of conservation
Commented [S73]: Please be consistent, if its correctly
abbreviated like this, rather use this abbreviation
throughout the document
NEMBA vs NEM:BA (No 10 of 2005) double check the
correct abbreviation and use it consistently throughout the
document

concern from NEMBAs TOPS list will be barcoded to create a library reference that can Commented [S74]: Use the correct abbreviation

help in with plant conservation.

Hartvig et al., (2016) reported added that correct identification for the threatened species

whose trade is regulated by Convention on International Trade of Endangered Species

(CITES ) is essential for the enforcement of laws and regulation as well as future

conservation of species. Vernooy et al., (2010) addedreported that understanding the

science of taxonomy is key to understanding and monitoring biodiversity. It can act as a

guidance to differentiate between legal and illegal species in trade (WWF, 2006).
LIST OF

REFERENCES Commented [S75]: I think your referencing should be

numbered, please check your department referencing
guide.
Department of Environmental Affairs and Tourism. (1998). Convention on Biological

Diversity. South African National Report on the convention on Biological Diversity. Pg. Commented [S76]: Most of you intext references are not
included in your List of References.
50. Please make sure that every reference source of an intext
reference is included in your list of References

Davis, J. and Mc Coy, J. (2000). Commercial Goldenseal Cultivation. NC State

Extension Publication. Available from: https://content.ces.ncsu.edu/commercial- Field Code Changed

goldenseal-cultivation. [Accessed on 03/08/2017] Formatted: No underline, Font color: Auto

Ethekweni Municipality. (2009). Our Bio Diverse City. Available from Formatted: Line spacing: Double

(http://www.durban.gov.za/City_Services/development_planning_management/environ

mental_planning_climate_protection/Publications/Documents/Our%20Biodiverse%20Cit

y.pdf) Accessed on [ 14 April 2017]. Pg. 9.

Germishuizen, G. and Meyer, N.L. (2003). Plants of Southern Africa: an annotated

checklist. Strelizia Strelizia. (14). Pg. iv.

Ethekweni Municipality. (2009). Our Bio Diverse City. Available from

(http://www.durban.gov.za/City_Services/development_planning_management/environ Formatted: Default Paragraph Font, Font: (Default)

+Body (Calibri), 11 pt
mental_planning_climate_protection/Publications/Documents/Our%20Biodiverse%20Cit

y.pdf) Accessed on [ 14 April 2017]. Pg. 9.

Hartvig, I., Czako, M., Kjaer, E. D., Nielsen, LR and Theilade, I. (2015). The use of DNA

barcoding in Identification and conservation of Rosewood (Dalbergia spp.). PLos ONE.

(9). Pg.2.

IUCN. (2017). Invasive species. Available from https://www.iucn.org/theme/species/our-

work/invasive-species) . [Accessed on 10 July 2017].

Lawson, K. and Vines, A. (2014). The global impacts of Illegal Wildlife Trade. The

Royal Institute of International Affairs. Pg.3.

Mutia, T.M. (2009). Biodiversity Conservation. Geothermal Development Company

Limited. Pg.4. Formatted: Font: Italic

National Environmental Biodiversity Act. (March 2015). Government gazette no.38600.

(597). Pg.27.

Pandy, R., Jha,D.K., Thapha, N., Pokharel, B.R. and Aryal, N.K. (2014). Forensic

Wildlife Parts and their Product Identification and Individualization using DNA

Barcoding. The Open Forensic Science Journal. (7). Pg.6-13.

Paterson, A. R. (2009). Legal framework for protected areas: South Africa. Pg.20.

Reid, W.V. (1998). Biodiversity hotspot. Elsevier Science. (3). Pg.275.

Slingeberg, A., Braat, L., Van der Windt, H., Rademaekers, K., Eicheler, L. and Turner,

K. (2009). Study on understanding the causes of biodiversity loss and the policy

assessment framework. European Commission Directorate-General for Environment.

South African National Biodiversity Institute. (2017). e-flora of South Africa. Available

from ( http://www.sanbi.org/biodiversity-science/foundations-biodiversity/biosystematics-

collections/e-flora) Accessed on [31/07/2017].

South African Biodiversity Institute. (2017). Threatened Plant Conservation. Biodiversity

advisor. Available from ( http://biodiversityadvisor.sanbi.org/planning-and-

assessment/plant-conservation-strategy/target-7/) [ Accessed on 31/07/2017].

Strydom, H. A. and King, N.D. (2009). Environmental Management in South Africa. 2nd

ed. JUTA law. Pg. 372.

Swingland, I. (2002). Capturing carbon and protecting biodiversity. The Royal Society.

Pg.

Taylor, J.E., Lee, S. and Crous,P.W. (2001). Biodiversity in the Cape Floral Kingdom:

Fungi occurring on Proteaceae. The British Mycological Society. Pg.1480.

Tschakert,P., King, B. Baum, S. and Wang, C. (2017). Threats to Biodiversity.

Pennstate. Pg.1.

Tshisikwawe, M.P. and Van Rooyen, M. W. (2012). Population biology of Brackenridgea

zanguebarica in the presence of harvesting. Journal of Medicinal Plants Research. (6).

Pg. 5748-5756.
Twine, W., Moshe, D., Netshiluvhi, T. and Siphungu, V. (2003). Consumption and

direct-use values of Savanna bio-resources used by rural household in Mametja, a

semi-arid area of Limpopo province, South Africa. South African Journal of Science.

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