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The influence of processing on the microbial risk associated with Rooibos

(Aspalathus linearis) tea

Article  in  Journal of the Science of Food and Agriculture · December 2014

DOI: 10.1002/jsfa.6719

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Received: 14 August 2013 Revised: 23 April 2014 Accepted article published: 30 April 2014 Published online in Wiley Online Library: 2 June 2014

(wileyonlinelibrary.com) DOI 10.1002/jsfa.6719

The influence of processing on the microbial

risk associated with Rooibos (Aspalathus
linearis) tea
Pieter Gouws,a* Toni Hartelb and Rudean van Wykb

Abstract
This review discusses the influence of processing on the microbial risk associated with Salmonella in Rooibos tea, the
identification of Salmonella and preventative and control measures to control microbial contamination. Rooibos tea, like other
plant products, naturally contains a high microbial load. Downstream processing steps of these products usually help in reducing
any contaminants present. Due to the delicate flavour properties and nature of Rooibos, gentle processing techniques are
necessary for the production of good quality tea. However, this has a major influence on the microbiological status of the
product. The presence of Salmonella in Rooibos is poorly understood. The ubiquitous distribution of Salmonella in the natural
environment and its prevalence in the global food chain, the physiological adaptability, virulence of the bacterial pathogen
and its serious economic impact on the food industry, emphasises the need for continued awareness and stringent controls at
all levels of food production. With the advances of technology and information at hand, the processing of Rooibos needs to be
re-evaluated. Since the delicate nature of Rooibos prohibits the use of harsh methods to control Salmonella, alternative methods
for the steam pasteurisation of Rooibos show great potential to control Salmonella in a fast, efficient and cost-effective manner.
These alternative methods will significantly improve the microbiological quality of Rooibos and provide a product that is safe
to consumers.
© 2014 Society of Chemical Industry

Keywords: Rooibos tea; microbial risk; Salmonella; fermentation; aspalathus linearis

INTRODUCTION safe to drink and of good export quality. Internationally and in

For 46 million South Africans, the agricultural and food process-
ing industries are of main importance. These industries contributes
approximately R124 billion to South Africa’s Gross Domestic Prod-
uct (GDP) each year. The South African agricultural sector provides
a host of investment and export opportunities in a number of dif-
ferent sectors whereby the country’s main crop exports include
wine, wheat, fruit, tea, tobacco, groundnuts and sugar.1 The South
African beverage market, including alcoholic, non-alcoholic, dairy
and powdered beverages, has also contributed significantly to the
gross income of the country.
In recent years the non-alcoholic beverage industry has gained
particular interest due to the consumer drive towards health
and wellness in South Africa and on a global scale. This trend
has therefore led to the worldwide demand for alternative and
healthier choices. As a result, the herbal tea market has flourished.
South Africa produces 25% of the world’s tea, of which Rooibos tea
and Honey Bush tea have gained tremendous popularity for their
unique taste and aroma.2
The first exports of Rooibos were in 1955 (524 tons), but it is only
during the past 14 years that the international market demand
steadily grew from 750 tons in 1993 to 7200 tons in 2007. In 2007
total production of Rooibos, including unfermented Rooibos, was
in excess of 14 000 tons, with Germany the major international
market (53%), followed by the Netherlands (11%), UK (7%), Japan
(6%) and USA (5%).2
Hence, due to the large consumption of Rooibos tea worldwide,
South Africa microbiological standards have been enforced which
Rooibos export companies have to adhere to. Although their
regulations differ in some aspects, all have a zero tolerance for the
presence of Salmonella spp. Salmonella is a serious food-borne
pathogen capable of causing salmonellosis, a gastrointestinal
disease.3
However, the literature available on the microbiological quality
of South African herbal tea is both minimal and dated. More
studies are therefore needed to elucidate the presence of microbial
contaminants in Rooibos tea.
This review discusses the influence of processing on the micro-
bial risk associated with Salmonella in Rooibos tea, the identifica-
tion of Salmonella and preventative and control measures to con-
trol microbial contamination.
ROOIBOS TEA
Rooibos (Aspalathus linearis) is a shrub-like leguminous bush,
native to the Cedarberg Mountains in the Western Cape region
of South Africa, where it is extensively cultivated within this area
∗
Correspondence to: Pieter Gouws, Department of Food Science, University of
Stellenbosch, South Africa. E-mail: pgouws@sun.ac.za
a Department of Food Science, University of Stellenbosch, South Africa
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a need arises for the production of tea that is microbiologically b Department of Biotechnology, University of the Western Cape, South Africa

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 www.soci.org
Figure 1. The location of the Cederberg area where the Rooibos is cultivated.6
for its commercial use as an herbal infusion material (Fig. 1).4 The
genus Aspalathus consists of over 200 species, which occur only in
South Africa.5
Rooibos has been traditionally used for medicinal purposes to
treat asthma, colic, eczema, headache, nausea and mild depres-
sion. The tea is rich in polyphenols, is caffeine free and con-
tains a rare source of the dietary dihydrochalcones aspalathin and
nothofagin.4 Most polyphenols have shown antimicrobial poten-
tial; however, their activity is dependant on the bacterial species
and type of polyphenols present. Polyphenols can inhibit growth
of Clostridia and Helicobacter pylori but have not shown antimicro-
bial action against intestinal lactic acid bacteria.7
Animal models have also shown Rooibos to possess potent
antioxidant activity, i.e. it is a scavenger of active oxygen species
which adversely affect human health,8 as well as antimutagenic9
and hypoglycaemic action.10 The strong antioxidant effect of Rooi-
bos has led to the use of the plant as an ingredient in cosmetic
products,11 which is the first non-beverage application of Rooibos.
Furthermore, Rooibos is also marketed in its unfermented form
known as Green Rooibos. It was first produced in 2001 to meet
the demand of consumers in need of a product with a higher
antioxidant potential. Unfermented Rooibos currently accounts for
1% of total sales of Rooibos sold worldwide.2
The processing of Rooibos tea
Rooibos is harvested during the months of January to April by
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cutting the plant approximately 45 cm from the ground2 and
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P Gouws, T Hartel, R van Wyk
placed into bundles for further processing. The tea is then passed
through a commercial cutting machine and cut into the desired
length (∼3–4 mm). Thereafter the moisture content of the tea is
increased by the addition of water followed by a bruising step. The
bruising of the tea is necessary for the release of polyphenols and
oxidative enzymes needed to speed up the fermentation process.
At this stage, the Rooibos is still green in colour.
The tea is left to ferment in heaps at ambient temperature for
approximately 12–14 h. As the fermentation progresses, the heaps
reach a temperature of 38–42 ∘ C depending on environmental
conditions.12 The fermentation heap is turned over several times
to ensure adequate aeration that is required for oxidation. The
fermentation is an essential step within the processing of Rooibos
as it imparts its characteristic brick-red colour, taste and aroma.
Fermentation is essentially an oxidative process carried out by
endogenous plant enzymes that oxidise chemicals present in the
plant such as polyphenols.2 The fermented tea is then sprayed
onto cement floors/tea courts, using tractors, where it is allowed to
sun dry for ∼6 h and thereafter stored in bulk. It is worthy to note
that the tractors and boots used by farm workers are only used on
the tea courts to prevent cross-contamination. Subsequently, prior
to the packaging of the tea, the product is pasteurised.
The pasteurisation of the tea is carried out using steam. The tea
leaves reaches a temperature of 85–92 ∘ C for 2.5 min to reduce any
microbial contaminants present and then sent off to a set of sifts
to remove excess moisture. The drying of the tea is essential since
the low water activity will prevent the growth of moulds. Lastly, the
J Sci Food Agric 2014; 94: 3069–3078
Effect of processing on microbial risk in Rooibos tea www.soci.org

Addition of water
and bruising Table 1. Microbiological guidelines for Rooibos tea in Europe and
South Africa
Harvest and Transportation Fermentation
cutting of Harvest
34-42°C Microbial
12-14h
contaminant Europe* South Africa†

Aerobic plate count 1.0 × 108 CFU g−1 7.5 × 104 CFU g−1
Pasteurisation (bulk); 1.5 × 105
85-92°C for 2.5 min Bulk Storage Sun dried for 6 h CFU g−1 (retail)
Yeasts 1.0 × 106 CFU g−1 –
Moulds 1.0 × 106 CFU g−1 –
Escherichia coli 1.0 × 104 CFU g−1 2.0 × 101 CFU g−1
Drying and Salmonella Absent in 5 × 25 g 0
Packaging Final Product
* Guidelines extracted from European Herbal Infusion Association

Figure 2. Schematic overview of the processing of Rooibos tea. Microbiological Guidelines.

† Regulations governing microbiological standards for foodstuffs and
related matters, Foodstuffs, Cosmetics and Disinfectants Act of South
tea is sent to a large container for bulk packaging. The processing Africa, 1972.
of Rooibos tea is shown schematically in Fig. 2.

Microbial risks associated with Rooibos tea

Rooibos tea, like other plant products, naturally contains a high
microbial load. Downstream processing steps of these products
usually help in reducing any contaminants present. Due to the del-
icate flavour properties and nature of Rooibos, gentle processing
techniques are necessary for the production of good quality tea.
However, this has a major influence on the microbiological status
of the product.
The long fermentation times, high moisture content and temper-
atures during production provides optimum conditions for bacte-
rial and mould growths. In addition, the extended exposure of the
tea to open areas further provides the opportunity for contami-
nation by airborne bacteria and spores. A study conducted by Du
Plessis and Roos13 illustrated the effect of fermentation on micro-
bial growth. Their data indicated that the colony count of Rooibos
tea artificially spiked with Escherichia coli and Salmonella enteri- Figure 3. Scanning electron micrograph of Salmonella Typhimurium.21
tidis, increased from 104 CFU g−1 to 108 CFU g−1 after fermentation
for both microorganisms tested. peritrichously flagellated, Gram-negative and facultative anaer-
The presence of microbial organisms in food can be responsible obic bacteria. They are able to carry out mixed acid fermenta-
for both spoilage and health hazards.14 It is therefore important tions producing mainly lactate, acetate, succinate, formate and
to minimise such contamination of food and the consequences of ethanol.3 Salmonella grows between pH 6.6 and 8.2 with a neutral
microbial growth. Microbiological standards have therefore been pH optimum. The bacterium is unable to tolerate high salt concen-
established in South Africa (Foodstuff, Cosmetics and Disinfectants tration and growth is inhibited below a water activity (aw ) of 0.94.
Act)15 and on the European market [European Herbal Infusion
Association (EHIA)].16
These standards serve as guidelines for the accepted level of a Pathogenicity of Salmonella
microorganism that may be present per gram of Rooibos tea. As Salmonella is an important food-borne pathogen and is one of the
shown in Table 1, the South African standard enforces a more strin- leading causes of food poisoning worldwide. The main clinical syn-
gent acceptance level as compared to Europe. However, although dromes associated with Salmonella infection are enteric (typhoid)
the guidelines differ between Europe and South Africa, both coun- fever and gastroenteritis. Enteric fever is a systemic illness that
tries have a zero tolerance for the presence of Salmonella in Rooi- results from infection with the exclusively human pathogens S.
bos. typhi and S. paratyphi. Without treating enteric fever, mortality is
In view of the fact that the Rooibos is prepared by adding boiling 12–30%.18
water to the tea, the guidelines are acceptable. The heat from the The non-typhoidal strains of Salmonella are the causative agent
boiling water kills a large number of the microorganisms present of salmonellosis, a gastrointestinal disease commonly associated
in the tea thereby ensuring that the hot beverage is perfectly safe with contaminated foods such as raw poultry products, eggs,
for consumption.17 Manufactures should therefore clearly state on pork and processed meats.19 Salmonellosis is caused by over 2000
their packaging correct preparation instructions. serovars; however, the most frequently reported one from humans
is Salmonella enterica serovar Typhimurium (Fig. 3).
Salmonella usually has an incubation period of 1–7 days where
SALMONELLA SPP. thereafter the disease presents itself via vomiting, nausea, diar-
The genus Salmonella belongs to the family Enterobacteriaceae, rhoea and abdominal pain which typically persist for 2–5 days but
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also known as enteric bacteria. Salmonella spp. are rod-shaped can last several weeks. The bacteria are able to invade and multiply

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 www.soci.org
within the intestinal mucosa where they produce an enterotoxin
and cytotoxin that can destroy epithelial cells.20
Salmonella in herbal tea
There is a noticeable lack of research concerning the microbiolog-
ical quality of herbal tea, especially locally produced tea. In a study
conducted by Du Toit et al.14 on honeybush, another well-known
South African tea, two thermophilic moulds, a thermo-tolerant
mould, Rhizomucor pusillus, and five endospore-forming species of
the genus Bacillus were isolated.
The data indicated that R. pusillus was the predominant microbial
contaminant; however, the honeybush was not tested for the
presence of Salmonella. The author suggested that the low curing
temperatures (40–50∘ C) of the honeybush were ineffective in
killing the microbial contaminants present. No contaminants were
present when the curing temperature was increased to > 60∘ C. In
addition, the higher temperatures did not affect the organoleptic
quality of the product.
Furthermore, a study conducted by Foote,22 examined the
microbiological status of chamomile tea. Significant amount of
yeasts and moulds were present on the chamomile; however, no E.
coli, Staphylococcus aureus or Salmonella spp. were detected. Simi-
larly, the study proposed that the microbial load was an indication
of the natural microflora of the plant as well as the handling of
the tea.
The presence of Salmonella in Rooibos is poorly understood. The
literature available is also limited and outdated. Studies have iden-
tified the presence S. enteritidis and S. lindrick from the fermenta-
tion heaps (after 23 h) during processing. S. greiz was also isolated
from the product after a 2 min steam pasteurisation procedure. The
steam pasteurisation treatment was highly effective in reducing
the coliform count but failed to eliminate the Salmonella present.
The results suggest that high levels of Salmonella must have been
present prior to pasteurisation.13
Similarly, a study conducted by Swanepoel23 detected the pres-
ence of 46 different serovars isolated from Rooibos. None of the
identified serovars was of the highly pathogenic Salmonella such S.
Typhimurium or S. paratyphi. The author suggests that the sources
of contamination could be of environmental origin such as lizards
and/or farm workers’ hands.
The use of Rooibos, with its broad appeal and use by a wide range
of population groups, including high-risk individuals, creates a
safety concern and merits a microbiological evaluation of the
product.
Sources of contamination
Salmonella is widely distributed in the environment; however,
most Salmonella serovars are associated with animal species such
as sheep (S. ser. Abortus ovis), pigs (S. ser. Cholerae suis), poul-
try (S. ser. Gallinarum), horses (S. ser. Abortus equi) and cattle
(S. ser. Dublin). Animals infected after exposure to infected ani-
mals, feed or environmental conditions excrete Salmonella by fae-
cal shedding. The faecal or intestinal contamination of carcasses is
the primary source of human food-borne infections.20
Salmonella can be further disseminated into the environment by
animal carriers. Salmonella has been isolated from various animal
species such as birds,24 muscoid flies,25 reptiles26 and mice.27 These
carriers play an important role in the epidemiology of Salmonella
contamination especially in food production warehouses/plants.
These animals may pass freely through the plantation and excrete
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Salmonella into its surroundings.
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P Gouws, T Hartel, R van Wyk
In a study to examine mice as carriers of Salmonella, observed
that up to 104 colony forming units per gram (CFU g−1 ) of
Salmonella were detected in a single dropping.27 Furthermore,
Salmonella can also survive in aquatic environments such as
surface and waste waters which further serve as a source of
contamination when used on farms or food plantations.28
The possible presence of Salmonella contaminants in Rooibos
tea may therefore be traced back to various sources such as
the soil the plant was grown in, irrigation water and water used
during fermentation, contaminated equipment due to animal
carriers etc. Because Salmonella is able to infect a large number
of animal species, identification of the source of environmental
contamination may not always be possible.
Even so, survival of Salmonella in the soil and in water provides
the bacterium with an increased probability of infecting a new
host.29 Salmonella does not multiply significantly outside its nat-
ural environment; however, the bacterium can survive in soil if the
temperature, humidity and pH conditions are favourable.13
The ubiquitous distribution of Salmonella in the natural environ-
ment and its prevalence in the global food chain, the physiological
adaptability, virulence of the bacterial pathogen and its serious
economic impact on the food industry, emphasises the need for
continued awareness and stringent controls at all levels of food
production.
IDENTIFICATION OF SALMONELLA SPP.
Conventional and molecular detection methods
The detection of Salmonella by conventional methods includes
a non-selective primary enrichment step, followed by a selective
secondary enrichment step, isolation on a selective agar media
and a preliminary biochemical confirmation test.30 Conventional
culturing techniques are often labour intensive, time consuming
and requires 4–6 days to produce results. Furthermore, the results
may also show poor sensitivity and low level contamination.31
Biochemical testing is also used to confirm the presence of
Salmonella in samples. These tests often make use of the API
20E test, a diagnostic strip capable of detecting 20 biochemical
reactions within the Enterobacteriaceae family.32 In recent years,
molecular based methods such as polymerase chain reaction (PCR)
have been successfully used to detect the presence of food-borne
pathogens including Salmonella.19
The PCR used for the detection of Salmonella is based on the
amplification of a 429 base pair region specific to Salmonella spp.
using the ST11 and ST15 primers. PCR allows for a more specific
and sensitive detection method as compared to conventional
techniques alone. Furthermore, the simplicity and rapidity of PCR
makes it a preferred choice.31
However, according to Myint et al.33 it was observed that PCR
failed to detect a positive sample when no pre-enrichment was
conducted. Furthermore, the sensitivity of the assay was also
considerably decreased when only a primary enrichment step
was performed. These results illustrate the need for conventional
methods in conjunction with molecular techniques to provide
accurate and conclusive results.
Typing methods
Although the standard PCR technique offers a high degree
of specificity, the assay is limiting with regards to Salmonella
species/strain characterisation. The ability to characterise specific
strains of Salmonella provides valuable insight into the primary
J Sci Food Agric 2014; 94: 3069–3078
Effect of processing on microbial risk in Rooibos tea
Figure 4. Phylogenetic tree of the evolution of Salmonella spp. within
closely related families.38
sources of contamination within the processing of Rooibos.
This information will therefore lead to the improvement of the
processing conditions and microbiological quality of the tea.
There are a number of ways Salmonella isolates are charac-
terised on a species level, either using phenotypic or geno-
typic methods.34 Phenotypic typing techniques are based on the
expression products of particular genes that may be present
in the different strains of bacteria. Conversely, genotypic tech-
niques/molecular typing identify differences in the nucleotide
sequence of the genome to type bacteria, and are largely indepen-
dent of gene expression.35
Phenotypic methods include phage typing, antimicrobial sus-
ceptibility testing and serological tests. Each method may be
advantageous over the other with regards to specificity and repro-
ducibility; however, the most common phenotypic method used
to differentiate Salmonella isolates is by serotyping.34 Moreover,
numerous molecular typing techniques have also been applied
to characterise Salmonella such as restriction enzyme-based,
PCR-based and sequencing-based methods.
Serotyping
Over 2500 serovars of Salmonella have been identified are tax-
onomically divided into two species, i.e. Salmonella enterica and
Salmonella bongori of which each contains sub-species differentia-
tion. The serology of Salmonella is based on the Kauffmann–White
scheme, which differentiates Salmonella serovars by the surface
antigen differences of somatic (O) and flagellar (H) antigens.36
A suspension of Salmonella is mixed and incubated with a
panel of antisera specific for a variety of O and H surface anti-
gens. Specific agglutination profiles are then generated and used
to determine the serotype present.34 However, this method is
labour-intensive, expensive, complicated and time-consuming.37
Also, it does not provide a basis for investigating evolutionary
genetic relatedness among strains (Fig. 4).36
Molecular typing
In recent years molecular typing assays have been employed to
overcome the problems associated with phenotypic methods.
A particularly useful PCR-based molecular typing method for
Salmonella is repetitive element PCR (Rep-PCR). This technique
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www.soci.org
detects non-coding repetitive sequences within the genome
of the microorganism and generates type specific banding
patterns.35
In particular, enterobacterial repetitive intergenic consensus
(ERIC)–PCR, a type of Rep-PCR, is designed specifically for enteric
bacteria. ERIC elements (highly conserved regions within genome)
are amplified using ERIC 1 and ERIC 2 primers. Following PCR, the
amplicons are separated by gel electrophoresis and the banding
patterns are compared to one another. Differences in the sizes
of the amplified fragments produce different banding patterns
(Fig. 5).
One of the major advantages of this set of techniques is that
the results can be obtained in a relatively short amount of time
with a relatively high discriminatory value and the techniques are
relatively easy to perform. According to a study conducted by Lim
et al.37 comparing four molecular typing methods, the ERIC-PCR
technique could differentiate the 57 Salmonella strains into 50
patterns well enough to be used for molecular typing.
PREVENTION AND CONTROL
Alternative methods to control Salmonella
Reduction of the number of microorganisms in Rooibos tea is
difficult due to the delicate nature and flavour properties of the
plant. Less abrasive methods are therefore necessary to control
microbial contamination. Currently, Rooibos tea farmers use steam
pasteurisation whereby water is heated to approximately 180∘ C to
generate steam. The Rooibos is then exposed using a conveyor
system, to the steam for 2.5 min and reaches a temperature of
85–92∘ C.
Steam pasteurisation treatment is efficient in reducing the
microbial load on Rooibos as well as significantly reduces the col-
iform count. However, the presence of Salmonella after pasteuri-
sation as observed by Du Plessis and Roos13 warrants the need for
alternative decontamination methods. In addition, steam pasteuri-
sation can also reduce the antimutagenic potential of the tea40 as
well as result in the loss of volatile oils17 thereby compromising the
organoleptic quality of the product.
Other, less abrasive and more competent methods are available,
such as the use of radiation, Salmonella specific bacteriophages
and ozone treatment.
Food radiation
Food irradiation involves the use of high-energy radiation in any
of three approved forms, i.e. gamma rays, X-rays, or electron
beams. Gamma rays can be generated by either of two approved
radionuclide sources, cobalt-60 or caesium-137, whereas X-rays
and electron beams are generated electrically.
Doses of radiation used in food processing are measured in units
of grays (Gy) or kilograys (kGy) where less than 1 kGy (low dose) is
used for disinfestation and the extension of shelf life, 1–10 kGy for
pasteurisation of meats, poultry and other foods, and more than
10 kGy for sterilisation.41
Radiation is also referred to as a cold pasteurisation as this tech-
nology can reduce the microbial load of a food product with-
out significant changes in temperature and chemical and phys-
ical composition of the product.42 Furthermore, this technology
can also be applied as a final treatment step after packaging to
ensure the product is free from contaminants until it reaches the
consumer.43
The use of radiation is applied in various sectors within the food
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industry such as meat, poultry, fruits and vegetables, spices and
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A B C

Figure 5. (A) Schematic representation of the repetitive element-based PCR. Hatched sections represent the primers complementary to the repetitive
sequences in the bacterial genome. (B) PCR products with different molecular sizes. (C and D) Gel electrophoresis: PCR products are separated by sizes
resulting in strain-specific fingerprints.39

herbs and is accepted by international health organisations such

as the US Food and Drug Administration (FDA) and World Health
Organization (WHO).
The use of radiation in the microbial decontamination of tea
herbs has gained particular interest in recent years. According to
Mishra et al.,44 the use of gamma radiation was successfully applied
in the decontamination of Camellia sinensis, commonly known as
black tea. The radiation dose of 5 kGy was observed to effective
and had no effect on the chemical and physical characteristics of
the tea. Likewise, in a similar study conducted by Thomas et al.,45
irradiation resulted in the removal of microbes present in the tea
and increased its shelf life without affecting the quality of the
product. Figure 6. Electron micrograph of Salmonella phage P7.48
To date, no published studies have been conducted on the use
radiation on Rooibos specifically. However, the effect on radiation also a useful method due to its speed, relative simplicity, and
on the quality of the product is dependent on the composition cost-effectiveness.49 More importantly, this technology can be eas-
of the food product itself therefore may not be applicable to ily applied to the processing of Rooibos tea. The phages are simply
all types of tea on the market. Further studies are therefore added to the water used during fermentation resulting in a prod-
needed to elucidate the potential use of radiation in the microbial uct that is both safe and of export quality (Fig. 6).
decontamination of Rooibos.
Ozone treatment
Salmonella-specific bacteriophages Ozone is a strong oxidant and potent disinfecting agent and
Bacteriophages represent a group of viruses that infect and repli- is generated by ultra-violet radiation (188 nm wavelength) and
cate in bacteria. Phages are extremely host-specific and are able corona discharge methods. Ozone destroys microorganisms by
only to infect specific species or even strains of bacteria. Salmonella the progressive oxidation of vital cellular components. Ozone
specific bacteriophages have been reported as biocontrol mea- degradation of the unsaturated lipids in the cell envelope results
sures in a number of food products such as broiler chicken,46 seed in cell disruption and subsequent leakage of cellular contents.50
sprouts,47 and cooked and raw meat products.48 All studies agreed The use of ozone is widely applied in the food industry including
that with the correct concentration and cocktail of phages, these increasing the shelf life of fruits and vegetables. The antibacterial
viruses can be successfully used as control measure for food-borne effects of ozone have been documented for a wide variety of
pathogens. organisms, including Gram-positive and Gram-negative bacteria
The extreme specificity of phages renders them ideal can- as well as spores and vegetative cells.51 However, further analysis
didates for applications designed to increase food safety dur- is needed to test the efficiency and viability of ozone against
3074

ing the production process. Furthermore, phage treatment is Salmonella spp. in Rooibos tea.

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Table 2. Commercial significance of metabolic products of lactic acid bacteria (adapted from Holzapfel et al.)54

Metabolite Beneficial Deleterious

Lactic acid Preservation; sensory improvement; enhancement Acidification

of digestion and nutrient uptake
Acetic acid Aroma Off-taste
Diacetyl/acetoin Aroma (dairy products) Off-taste (beer)
CO2 Preservation; taste enhancement Gas production (blowing/bloaters)
H2 O2 Preservation Discolouration; greening
Biogenic amines – Health (allergies)
Slime Stabilisation (e.g. yoghurt) Sensory
Methane thiol. H2 S Aroma Sensory
Growth factors Aroma; nutritional value Clostridia (gas); yeasts
Bacteriocins Preservation (inhibition of closely related bacteria) Health; inhibition of beneficial lactic acid bacteria
Wide-spectrum antimicrobials Inhibition of pathogens and spoilage Possible allergies; resistance of intestinal
microorganisms microorganisms

Table 3. Practices for the processing of Rooibos

Production step Practices

Cultivation • Herbal materials should not be cultivated in soils contaminated with, for example, sewage sludge, heavy metals,
pesticides, radio-elements and other industrial chemicals
• Water used for irrigation should be fit for the purpose, i.e. substantially free from contaminants, such as faeces, heavy
metals, agrochemicals
• No cattle should be allowed in the cultivation area
• No sewage sludge should be used for fertilisation

Harvesting • Harvesting should be carried out in dry, low humidity conditions to avoid mould growth
• Harvesting equipment and mechanical cutters should be clean and well maintained
• All containers used for primary collection of the crop when not in use must be kept in a dry place free from vermin and
inaccessible to farm and domestic animals
• The harvested crop should be protected from all types of pests (rodents, insects) and farm and domestic animals
• Water used on raw material must be free of contaminants

Drying • Areas used for drying crops should be well ventilated and never used for livestock
• The drying area should be constructed to protect the crop from birds, insects, rodents, farm and domestic animals
• There should be strict physical separation of harvested materials from waste containers
• Clearly marked waste bins should be provided, emptied daily and cleaned
• Dried crops should be stored as soon as possible for protection and to lessen the opportunity of pest infestation, as
well as to prevent ingress of foreign matter

Packing • Packaging materials should be stored in a clean dry place free from pests and inaccessible to animals
• The packed crop should be stored in a dry place away from the wall and off the ground and be protected from pests
and farm and domestic animals

Storage and transport • Packed dried crop should be stored in a dry, well-ventilated building, with minimal variation in diurnal temperature
• Shutter and door openings should be protected by wire screens to keep out pests, birds, and farm and domestic
animals. There should be appropriate pest-control measures, such as traps, electrical insect-control devices, as well
as measures for identifying an infestation
• For bulk deliveries the use of vented containers and transport vehicles is highly recommended to minimise mould risks
• The transport vehicles should be clean and in good condition
• Chemicals used as pesticides, fumigants etc., should be kept in a separate area

Equipment • Equipment used for the gathering, handling and processing of crops should be easily cleaned to minimise contami-
nation

Personnel and facilities • All persons having contact with raw materials should observe a strict level of personal hygiene
• Personnel must not be permitted to work in the herbal material handling area if they are known to be suffering from,
or carriers of, diseases likely to be transmitted through food, including diarrhoea
• Personnel with open wounds, sores, and skin infections should be transferred away from herbal material handling
areas until completely recovered
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 www.soci.org P Gouws, T Hartel, R van Wyk

Table 4. Hazard analysis critical control points (HACCP) plan for the production of Rooibos tea
Production step Possible hazard CCP/QCP Control measure Critical limit Monitoring procedure Corrective action

Delivery Soiled packs. QCP Visual inspection of Material and packs meet Visual inspection Rejection, sorting,
Cross-contamination from incoming deliveries requirements instructions to
other raw materials, and removal of packs supplier
infestation which do not meet
requirements
Mechanical Soil, stones, twigs, leaves, QCP Removal by hand and/or No obvious Visual inspection before –
precleaning other foreign bodies by sieves contamination and after removal
Cutting Metal fragments (broken QCP Inspect equipment No metal fragments in Magnets can be used Repair and replace
pieces of blade) regularly, observe the material when filling material damaged equipment.
maintenance into sacks, e.g. metal Install magnets
procedures detectors at a later
production stage
Fermentation Mould and bacterial growth No – – – Downstream
pasteurisation
Drying in the sun Inadequate drying, QCP Ensure proper air e.g. Maximum layer Check layer thickness, Re-dry, sorting out
composting, formation of circulation, optimise thickness. e.g. Turning drying time and if
mould drying conditions frequency (e.g. twice necessary
daily) Temperature temperature. Sensory
control Sufficient perception, visual
ventilation Humidity inspection, humidity
gauge
Sun-drying: pollution by QCP Use covering nets, No contamination Visual inspection Sorting out
animals fencing or provide a
roof
Metal fragments (broken QCP Inspect equipment No metal fragments in Magnets can be used Repair and replace
pieces of shovel or metal regularly, observe the material when filling material damaged equipment.
grids) maintenance into sacks, e.g. metal Install magnets
procedures detectors at a later
production stage
Bulk storage Infestation (insects, rodents, QCP Pack as soon as possible. No infestation Suitable control Sorting out, fumigate if
insect damage, Keep material measures (traps, necessary
contamination (by farm protected by using electric insect
animals and domestic wire mesh to cover eradicators,
animals, birds) shutters and monitoring for pests)
doorways
Mould and bacterial growth QCP Temperature and No obvious Humidity gauge/air Sorting out
humidity control contamination conditioner
Pasteurisation Faulty temperature gauge CCP Faulty equipment Pasteurisation must be Inspect pasteurisation Testing of final product
and timer should be repaired as carried out at correct equipment regularly
soon as possible temperature and
duration
Drying Mould growth QCP Spread in thin layers on Thickness. e.g. Turning Visual inspection Re-dry, sorting out
sieves, proper air frequency (e.g. twice
circulation daily). Temperature
control, sufficient
ventilation, humidity
Packaging Contaminated packaging QCP Removal of packs which Material and packs meet Visual inspection Rejection, instruction to
material do not meet requirements supplier
requirements.
Materials from
authorised supplier

CCP, critical control point; QCP, quality control point.

Lactic acid bacteria as biocontrol agents
As Salmonella has been shown to be present after pasteurisation,
as previously described, alternative methods which do not signif-
icantly affect the quality of the food product have become more
important in the food industry. The prospect of using lactic acid
bacteria (LAB) has become a very appealing topic within the scien-
tific community as these bacteria render many advantages during
food fermentation and also food preservation.52
The advantages of using LAB as starter cultures may not
only include industrial benefits such as lowering economic
expenses, enhancing the fermentation processes within the
industry, enhancing the properties of the food product,53 extend
shelf-life54 but also includes health-related advantages such as
3076
A study carried out by Yasui et al.55 found that two specific strains
of LAB enhanced the humoral immunity of humans, provided
protection against viral infections and enhanced cellular immu-
nity by inhibiting tumour production and mediating allergic
reactions. This observation, among other research, has substan-
tiated and encouraged the use of LAB as starter cultures for food
fermentation processes.
LAB are generally defined as Gram-positive, non-sporing bac-
teria which are non-aerobic but aero tolerant and which have
morphological characteristics of being cocci or rod-like in shape.56
LAB have been implemented in the industry for many years as
they are involved in spontaneous food fermentation processes.57
Dependent on the mode of fermentation the bacteria use,

being a positive enforcement within the human immune system.55 homo-fermentation or hetero-fermentation, the main product

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Effect of processing on microbial risk in Rooibos tea
formed is lactic acid as well as other organic acids (including acetic
acid), ethanol and bacteriocins (Table 2).58
Bacteriocins are described as any variety of protein (usually) that
are bacteriocidal or bacteriostatic most often specific to bacte-
ria closely related to the bacteriocin-producing microorganism.59
Bacteriocins produced by bacteria have become of great interest
within the microbiological community as they have been shown to
have an antagonistic effect on various bacteria. Such an example
is shown where 14 different strains of LAB from the genera Lac-
tobacillus, Leuconostoc, Pediococcus and Lactococcus inhibited the
growth of Listeria monocytogenes which is a pathogen that can
be isolated from the environment.60 Bacteriocin-producing LAB
isolated from retail cuts of meat were shown to have inhibitory
effects against Aeromonas hydrophila, Staphylococcus aureus and L.
monocytogenes.61 These bacteriocins include nisin, lacticin, ente-
rocin and many others of which nisin are the most abundantly
produced.62
Food safety systems
Prerequisite programmes
Even though microbiological guidelines are in place to control
the microbial load in Rooibos, it is important that producers
should continue to improve their cultivation and processing prac-
tises. The use of principles of Good Manufacturing Practice (GMP),
Good Agricultural Practice (GAP) and Good Handling Practice
(GHP), as prerequisite programmes, can help improve hygiene and
reduce microbiological contamination of the product. These prac-
tices include suitable premises for product manufacture, proper
transportation and storage of goods, sanitation and pest control,
hygiene of personnel, etc., as illustrated in Table 3.
Hazard Analysis Critical Control Points
In conjunction with these prerequisite programmes, the concept
of Hazard Analysis Critical Control Points (HACCP) should be imple-
mented. The execution of HACCP is particularly important as this
system helps to identify key steps within the processing of Rooi-
bos that need to monitored and kept under controlled conditions.
Jointly, these two concepts form a food safety system that can
minimise the microbial risk associated with the processing of Rooi-
bos. Although the tea is subject to further preparation with boiling
water preventative and control measures are crucial.
Table 4 is an example of a HACCP plan to facilitate the character-
istic processing of Rooibos to help minimise the risk of Salmonella
and other microbial contaminants.
CONCLUSION AND FUTURE PROSPECTS
With the global trend steering towards healthier food choices, it
is not unlikely that the demand for Rooibos will reach greater
heights. The high consumption of Rooibos tea worldwide there-
fore places great emphasis on the microbiological quality and
safety of the product, however little is known about its microbio-
logical status. To date, the scientific literature available has mainly
focused on the quality and health benefits of Rooibos rather than
its safety.
The fact that the conventional processing of Rooibos can poten-
tially promote the growth of Salmonella, and other microbial
contaminants, is of great concern to human health. Further inves-
tigation is therefore needed to isolate the sources of Salmonella
contamination within the processing of Rooibos, consequently
being able to implement preventative and control measures to
J Sci Food Agric 2014; 94: 3069–3078 © 2014 Society of Chemical Industry
www.soci.org
ensure the safety of consumers. The microbiological quality of the
tea will also have severe implications for the tea export industry
and manufacturers likewise, if the presence of Salmonella is not
dealt with accordingly.
With the advances of technology and information at hand,
the processing of Rooibos needs to be re-evaluated. Since the
delicate nature of Rooibos prevents the use of harsh methods to
control Salmonella, alternative methods to steam pasteurisation
of Rooibos show great potential to control Salmonella in a fast,
efficient and cost-effective manner. These alternative methods will
significantly improve the microbiological quality of Rooibos and
provide a product that is safe to consumers.
The advances in Rooibos production are inevitably hindered by
the lack of information available which is both minimal and dated.
Extensive studies are thus needed to investigate the need to isolate
the sources of microbial contamination and potentially improve on
the processing of Rooibos thereby reducing the risks and health
concerns involved.
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