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8]

Review Article

Challenges of Bovine Tuberculosis Control and Genetic

Distribution in Africa
Benjamin David Thumamo Pokam1,2, Prisca W. Guemdjom3, D. Yeboah‑Manu2, Elroy P. Weledji4, Jude E. Enoh1,5, Patience G. Tebid1, Anne E. Asuquo6
Departments of 1Medical Laboratory Science 3Public Health and 4Surgery, Faculty of Health Sciences, University of Buea, Buea, 5Laboratory of Endocrinology and
Radioelements, Medical Research Centre, Institute of Medical Research and Studies on Medicinal Plants, Yaounde, Cameroon, 2Bacteriology Department, Noguchi Memorial
Institute for Medical Research, University of Ghana, Accra, Ghana, 6Department of Medical Laboratory Science, College of Medicine, University of Calabar, Calabar, Nigeria

Abstract
Bovine tuberculosis (BTB), caused by Mycobacterium bovis (M. bovis); a member of the Mycobacterium tuberculosis complex (MTBC), is
a well‑known zoonotic disease, which affects mainly cattle. Control programs have either nearly or completely eliminated this disease from
domesticated animals in many developed countries. Its persistence in developing countries results from interactions between people, livestock
transhumance, and wildlife. In addition, deficiencies in preventive and/or control measures, poor sanitation, veterinary and slaughterhouse
services, and lack of political measures have been blamed. The proportion of human TB cases caused by M. bovis is most likely underestimated
since tests to distinguish between MTBC are seldom performed. Molecular techniques, especially spoligotyping, have helped to link human
and animal transmission. Several challenges in the control of M. bovis TB in Africa have been identified, and its eradication efforts require a
holistic approach. This review explores the challenges in the control efforts of BTB in Africa, as well as the impact of the genotyping evolution
and distribution of M. bovis in the continent and strategies to improve its control.

Keywords: Africa, bovine tuberculosis, control, genetic diversity, human transmission

Introduction the world.[3,4] Its persistence in developing countries results

Bovine tuberculosis (BTB) is a well‑known zoonotic disease
resulting primarily from infection with Mycobacterium bovis, a
member of the Mycobacterium tuberculosis complex (MTBC)
which consists of M. tuberculosis sensu stricto, Mycobacterium
africanum, Mycobacterium caprae, Mycobacterium
pinnipedii, Mycobacterium canettii, Mycobacterium microti,
Mycobacterium mungi, Mycobacterium orygis, Mycobacterium
suricattae, and the dassie bacillus.[1,2] BTB was previously
found worldwide; however, the introduction of control
measures including test and slaughter and pasteurization has
nearly eliminated this disease from domesticated animals
in many countries. Several European, Asian, or American
Nations are currently classified as BTB free or are underway
of eradication with adopted programs. BTB, however, is still
widespread in domestic animals or wildlife in parts of Africa,
Asia, the Pacific, and some Middle Eastern countries, as well
as some regions in the Americas and Europe. In 2017 and early
2018, the disease was declared present in 43% of reporting
countries and territories and is distributed in every region of
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from deficiencies in preventive and/or control measures, poor
sanitation, and health care.[5] It is classified by the Food and
Agriculture Organization of the United Nations (FAO) and
the World Organization for Animal Health (OIE) as a “List B”
disease; a category including “Transmissible diseases which
are considered to be of socioeconomic and/or public health
importance within countries and which are significant in the
international trade of animals and animal products.” (http://
www.fao.org/docrep/004/x3331e/X3331E01.htm).
Address for correspondence: Dr. Benjamin David Thumamo Pokam,
Department of Medical Laboratory Science, Faculty of Health Sciences,
University of Buea, P.O.Box, 63 Buea, Cameroon.
E‑mail: thumamo@yahoo.fr
ORCID: https://orcid.org/0000-0001-5828-8455
This is an open access journal, and articles are distributed under the terms of the Creative
Commons Attribution‑NonCommercial‑ShareAlike 4.0 License, which allows others to remix,
tweak, and build upon the work non‑commercially, as long as appropriate credit is given and
the new creations are licensed under the identical terms.
For reprints contact: reprints@medknow.com
How to cite this article: Pokam BD, Guemdjom PW, Yeboah-Manu D,
Weledji EP, Enoh JE, Tebid PG, et al. Challenges of bovine tuberculosis
control and genetic distribution in Africa. Biomed Biotechnol Res J
2019;3:217-27.
Received: 08.02.2019; Revision: 18.08.2019; Accepted: 20.09.2019; Published: 03.12.2019

© 2019 Biomedical and Biotechnology Research Journal (BBRJ) | Published by Wolters Kluwer - Medknow 217
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Pokam, et al.: BTB control in Africa
The genetic characterization of tubercle bacillus isolates
causing disease in humans has been extensively studied
contrary to those causing disease among wild and domesticated
mammals. [6] Despite the fact that very closely related
mycobacteria have adapted to causing TB disease in a
specific host species (some exclusively human), for example,
M. tuberculosis, M. africanum, M. canettii or rodent
pathogens (e.g. M. microti), interspecies transmission can occur
with a wide host spectrum as with M. bovis.[7,8]
Mycobacterium bovis Hosts and Transmission
Among the MTBC, M. bovis has the widest host range; it infects
mainly cattle, but cross‑infection of humans and a wide range
of both domestic and wild animals can occur. The pathogen can
be transmitted by aerosol and ingestion of infected carcasses.
The interactions between humans, livestock, wildlife, and the
consumption of unpasteurized contaminated milk and other
dairy products, especially in developing countries, puts people
at a greater risk of exposure to the disease. BTB manifests as
tubercles consisting of a necrotic caseous core surrounded by
granulomatous inflammation. These tubercles can be found
in any organ but are the most common in the lymph nodes,
particularly of the mammary gland and lungs.[9,10] Cattle shed
M. bovis in various secretions including the respiratory, feces,
milk, and urine, as well as vaginal secretions or semen. In most
cases, M. bovis is transmitted between cattle in aerosols in close
contact. Ingestion is the main route in calves that nurse from
infected cows. Cutaneous, genital, and congenital infections
have been seen but are rare. All infected cattle may not transmit
the disease, however, asymptomatic and anergic carriers occur.[3]
Eating meat from wildlife obtained by poaching/hunting which
are likely not to undergo veterinary inspection is another
potential source of infection to humans.[11] While disease
prevalence in cattle decreases, control efforts are sometimes
impeded by the passage of M. bovis from wildlife to cattle. In
epidemiological terms, the disease can persist in some wildlife
species, creating disease reservoirs.[12] Several animals have
been reported to be spillover hosts.[3] Although most mammals
may be susceptible, little is known about the susceptibility of
birds to M. bovis, as they are generally thought to be resistant
based on the experimental infection.[3]
Bovine Tuberculosis Control in Africa
Several control strategies such as tuberculin testing of
livestock, removal of positive reactors and milk pasteurization
have contributed to the very low risk of contracting zoonotic
TB in developed countries. The lack of control measures
among other factors contribute to the continued risk and
difficult eradication and spread of BTB in Africa. The impact
of noneradication will have global implications considering
the current trends in international trade and globalization. In
2016, 35 and 27 countries between January–June and July–
December, respectively, notified the disease in the African
region. None of the countries had either official vaccination,
218 Biomedical and Biotechnology Research Journal ¦ Volume 3 ¦ Issue 4 ¦ October-December 2019
treatment, or wildlife reservoir control policy, although 24
countries practiced some precautions at the borders. (www.
oie.int/wahis_2/public/wahid.php/Diseasecontrol/measures).
Wildlife reservoirs have been shown to hinder greatly the control
efforts of M. bovis.[12,13] The role of these reservoirs in M. bovis
disease dynamics has been established by various molecular
methods and guiding to some level the control measures,
especially in developed countries. It has also aided in tracing the
sources of infections that result from within‑herd transmission
and movement of livestock. Once BTB is established in a herd,
it spreads through aerosols, suckling, direct contact between
animals, and sharing of water and feed. The BTB infection in
maintenance hosts can persist through horizontal transfer in the
absence of any other source of M. bovis and can be transmitted to
other susceptible hosts.[14] Moreover, the comparison of the clonal
complex data from African regions indicates an intercountry
transmission associated with a country‑specific evolution, and the
clusters suggest the current transmission that occurs mainly within
cattle populations, less frequently between cattle and humans and
possibly between humans, pointing out the difficulty to develop
an efficient national control strategy of BTB in Africa.[15]
Challenges associated with Bovine Tuberculosis
Control in Africa
Diagnostic challenges of human Mycobacterium bovis
infection in Africa
Although studies in Africa have shown that a median of
2.8% (range 0%–37.7%) of all TB cases in humans is caused
by M. bovis,[16] these figures most likely underestimate the
true number of cases since differential diagnosis of MTBC
are seldom performed.[11] Several African settings still rely
on smear microscopy diagnosis of TB, with culture and
differentiation of M. tuberculosis species remaining limited.
The importance of the introduction of laboratory culture for the
specific identification of the mycobacteria in Africa has been
pointed out.[17] Several other factors might be associated with
the low prevalence of M. bovis in Africa; further contributing
to its underestimation. These include the generalized use of
glycerol in the culture media for tubercle bacilli isolation in
available facilities with such diagnostic capacity.[18] Human
disease caused by M. bovis and other species of the MTBC
are similar, although the anatomic site of M. bovis disease is
more often extrapulmonary. Therefore, identifying M. bovis in
extrapulmonary and respiratory samples would provide more
accurate estimates of M. bovis infections of human TB.[19]
It has been reported that approximately 13% of isolates were
more likely to be M. africanum and M. bovis rather than
M. tuberculosis.[20] The need for molecular characterization of
clinical isolates to ensure that correct estimates are made of the
true burden of infection due to M. bovis and M. africanum in
developing countries has been highlighted; M. africanum isolates
have substantial phenotypic heterogeneity with some strains
resembling M. bovis and others resembling M. tuberculosis.[21‑25]
This has led several authors to question the validity of the
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Pokam, et al.: BTB control in Africa
species designation.[26,27] In most cases, the available information
indicates the mere presence or absence of the TB disease but
does not provide any analysis of the spatial prevalence and
proportion of M. bovis infection in a specific region. Data on
human infection with M. bovis mainly stem from individual
research on molecular epidemiology or genetic diversity studies
of MTBC in Africa. Given the low‑sample sizes and the lack
of large‑scale, population‑based studies, available data are
insufficient to identify specific risk groups associated with M.
bovis infections.[16] The link between the population of M. bovis
strains causing human TB disease originating from a distant
population has been shown through spoligotype patterns.[28]
As advanced as the newly introduced Genexpert molecular
technique is, it is unable to differentiate the various species of
the MTBC. Consequently, patients infected by M. bovis who are
placed on standard treatment are likely to experience treatment
failure as a result of intrinsic pyrazinamide resistance.[29]
Transhumance control challenge
Several factors such as the variation in the ecological
zones (arid, semiarid, subhumid, humid, and highlands), the
differences in the production and movement of livestock
within the continent (pastoralism, “agro‑pastoralism,”
mixed farming, and intensive dairy farming), as well as the
macro‑ and micro‑climates affecting the stability of the agent
in the environment influence the distribution and transmission
of animal TB and thus its control in Africa. This affects its
prevalence between different areas.[30] About two‑thirds of the
African continent comprises arid or semi‑arid areas, and the
majority of rural African drylands are inhabited by livestock
keepers, who are either pastoralists or farmers; combining
rainfed agriculture with pastoralism.[31] Most of the pastoral
communities (Bedouins, Berbers, Maasai, Somali, Boran, and
Turkana) across the continent live in the semi‑arid grasslands
or arid deserts where rain‑fed agriculture is difficult, and the
effects of climate change observed in some subregions have
led to intensification of these movements. Drought generally
affects the life of pastoralists, and with the diminution of rains
and dried pastures, they are forced to move in areas where
the forage is available to avoid the starvation of cattle. The
mobility of animals and their herders is not only motivated
by the search for pasture and water but equally to avoid areas
affected by livestock disease or to engage in livestock trade.
The movements vary greatly in the distance and may take place
within a country or into bordering countries. The diversity of
the restriction fragment length polymorphism (RFLP) and/or
spoligotype patterns has been shown to reflect the extensive
internal movements of cattle belonging to pastoralists,[32]
and strains sharing common evolutionary origin have been
described between countries with cattle trading links.[20]
Nomadic or seminomadic pastoralists predominate across
the dryland Sahelian and Saharan belt that transects West,
North and the Horn of Africa, where the belt of aridity turns
southward to penetrate East Africa.[33] With cross‑border
mobility commonly practiced with some resultant occurrence
of friction and conflicts between pastoralists and local
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communities, countries such as Benin, Togo, Ghana, and Côte
d’Ivoire, like countries in the Sahel region, have become host
or transit countries,[34] and initiatives have been recorded in the
horn of Africa seeking to promote and document cross‑border
trade to stimulate regional growth, food security, and allow the
effective vaccination of livestock.[35]
Large variations in disease occurrence within different regions
of the same country have been reported in Africa.[36] High
prevalence rates of BTB have been described in areas where
cattle shared grazing and water as well as in areas where the
traditional management of livestock in transhumant herds
prevailed.[37,38] Under these often nomadic conditions, the
risk of exposure to M. bovis is increased significantly by
creating multiple herd contacts and increasing the total herd
size. The latter has also been suggested as a driver of the
disease prevalence.[39] Isolation of M. bovis from humans and
M. tuberculosis from livestock has pointed out transmission
between livestock and humans in the pastoral areas. [40]
Although some countries within the Economic Community
of the West African States have formulated and passed
legislation, these legal texts have not been put into practice.
Moreover, pastoralist development projects and programs
helping to improve transhumance practices by establishing
infrastructure along the livestock corridors (a strip of land
reserved for livestock passage to access pasture, a source
of water or other herd infrastructures such as a livestock
market, vaccination area, or livestock‑holding area) are still
insufficient.[41]
There have been initiatives seeking to promote cross‑border
trade and also document it, to stimulate regional growth and
food security and allow the effective vaccination of livestock.
Initiatives include Regional Resilience Enhancement Against
Drought, the Enhanced Livelihoods in Mandera Triangle/
Enhanced Livelihoods in Southern Ethiopia as part of the
Regional Enhanced Livelihoods in Pastoral Areas program in
East Africa, and the Regional Livelihoods Advocacy Project.[35]
Slaughterhouse to consumers and environmental security
challenge
Although postmortem inspection of carcasses is carried out at
the structured available slaughterhouse in Africa, low standard
of hygienic practices within its setting as well as unreliable
and inadequate meat inspection record have been reported.
This points out to the lack of thoroughness of the veterinary
staff incriminated with downstream consumers risk of BTB
infection.[42,43] Within the slaughterhouse, the diagnosis of BTB
can be made following the macroscopic detection at necropsy
of typical lesions, and postmortem inspection of carcasses
has been found to be insensitive for the detection of lesions.
To determine the significance of cattle that give a positive
reaction in diagnostic tests but do not have visible lesions due
to early infection, a bacteriological examination is necessary.
Although methods employing monoclonal antibodies and
DNA probes may be used to obtain a rapid identification,[44]
this is impractical in Africa due to lack of basic infrastructure.
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Risky behavior and poor attitudes and practices within African
slaughterhouse constitute itself the point of dissemination of
the disease, not only to workers but equally to the population
and environment. It has been observed that there are many risks
for those working within its vicinity that can create accidents
likely to promote zoonosis as BTB infection especially
because of lack of safety procedures observance (e.g. no
use of Personal Protective Equipment) leading to accidents
works such as injuries caused by the different sharp objects,
falls on wet, greasy, and slippery floor from handling raw
meat. Furthermore, good hygiene and sanitation are not
fully observed in slaughterhouse added to the lack of potable
water (the use of untreated water drawn from well polluted
with drain) put at‑risk workers to M. bovis infection. The
lack of proper sewage system leading to stagnant water,
environmental pollution, poor waste management, and
disposal contributing to pollution in slaughterhouse itself,
air, water, land in the vicinity [Figure 1], as well as delayed
disposal of animal carcasses such as abandoned horns often
heaped awaiting its sales and/or incineration have been
observed [Figure 2]. Slaughterhouses are a major source of
water and air pollution worldwide, and the environmental and
public health implications of unhygienic waste disposal has
been pointed out in Africa with recommendations for a safe
disposal following the international norms.[45]
Movement of people in and out of the slaughterhouse is not
controlled, and the lack of knowledge by the workers eating in
the slaughter hall and not respectful of personal hygiene and
sanitation all contribute to put at risk all those having access
to the slaughterhouse. Moreover, there are usually no medical
facilities to take care of any emergencies that may occur
while they are working. Interrupted power supply, inadequate
temperature checks and close monitoring of cold chambers for
proper meat preservation, as well as transportation of meat
in dirty car trunks or other available local transport systems,
constitute the poor system management of slaughterhouse in
Africa. Moreover, the deficiencies of veterinary services in
the rural and remote areas in developing countries constitute a
setback and thus likely to promote the spread of the infection.
Sometimes, animals are slaughtered outside approved
facilities, especially during transhumance, where some sick
animal can be sold to avoid the economic loss of herders.
Figure 1: Wastewater disposal from a slaughterhouse to close inhabited
environment
220 Biomedical and Biotechnology Research Journal ¦ Volume 3 ¦ Issue 4 ¦ October-December 2019
Furthermore, socioeconomic challenges faced by herders,
religious and various cultural practices as well as the inability
of the governments in Africa to compensate herders hinder
the test and slaughter policy, leading to the perpetuation of
disease transmission.
Molecular Epidemiology as a Contribution in
Bovine Tuberculosis Control in Africa
Host tropism and genetic evolution of Mycobacterium
bovis
The MTBC is characterized by 99.9% similarity at the
nucleotide level and identical 16S rRNA sequences but
differs widely in terms of their host tropisms, phenotypes, and
pathogenicity.[7,46,47] Gene expression differences between the
sequenced M. tuberculosis and M. bovis have been defined,[48]
and comparative analyses of the M. tuberculosis and
M. bovis genomes have revealed the basis for distinguishing
phenotypes such as the pyruvate requirement of M. bovis in
glycerol‑based media, or the reason for eugonic/dysgonic
colony morphology.[49] Contrary to earlier assumptions that
human TB evolved from the bovine disease by adaptation of
an animal pathogen to the human host,[50] new findings in the
light of genome sequencing indicate that TB first emerged in
humans and was subsequently transmitted to animals.[51] The
common ancestor of the MTBC is suggested to have emerged
from its progenitor perhaps 40,000 years ago in East Africa.
Some 10,000–20,000 years later, two independent clades
evolved, one resulting in M. tuberculosis lineages in humans,
while the other spread from humans to animals, resulting in
the diversification of its host spectrum and formation of other
MTBC members, including M. bovis.[8] Based on the high
sequence conservation in housekeeping genes, it has been
hypothesized that the tubercle bacilli encountered a major
bottleneck 15,000–20,000 years ago.[47] As the conservation
of the TbD1 junction sequence in all tested TbD1‑deleted
strains suggests a descent from a single clone, the TbD1
deletion is a perfect indicator that modern M. tuberculosis
strains that account for the vast majority of today’s TB cases
definitely underwent such a bottleneck and then spread
around the world, and the adaptation to animal hosts probably
Figure 2: Unsanitary environment and disposal of animal carcasses and
horns within an African slaughterhouse
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coincided with the domestication of livestock approximately
13,000 years ago.[8]
M. bovis has undergone numerous deletions compared to
M. tuberculosis. The genome sequence analysis of M. bovis
AF2122/97, isolated from cattle, revealed no new gene clusters
that were confined specifically to M. bovis, indicating that the
genome of M. bovis is smaller than that of M. tuberculosis.[52]
M. bovis seems, therefore, the final member of a separate
lineage represented by M. africanum, M. microti and M. bovis
that branched from the progenitor of M. tuberculosis isolates.
Successive loss of DNA may have contributed to clonal
expansion and the appearance of more successful pathogens
in certain new hosts.[7] Whole‑genome sequencing (WGS)
and comparative genomics have generated information
on the evolution and molecular basis of pathogenicity and
transmissibility. Although genomic information is more
available for M. tuberculosis infecting humans, little is
known about M. bovis, probably because of the limited
human‑to‑human transmission.[53] Widespread application of
WGS using approaches that more directly integrate it as an
additional epidemiological data to M. bovis will bring novel
and important insights into the dynamics of its spread and
persistence.[54]
Genotyping utilities and techniques for Mycobacterium
bovis detection
Different techniques have been developed for the genotyping of
M. bovis. Although the IS6110‑RFLP (classified with variable
number tandem repeat [VNTR] typing as the Tandem Repeated
Sequences) has been used as the gold standard for both human and
BTB over many years, its use in BTB is less common due to the
lower copy number of IS6110 in M. bovis.[55] Another technique
includes the spacer oligonucleotide typing (spoligotyping – a
nontandem‑repeated sequences) that identifies polymorphism
in the presence or absence of 43 specific DNA spacer units
in the direct‑repeat (DR) region in strains of the MTBC, with
M. bovis showing a characteristic absence of five spacers in
the 30 DR region (39–43).[56]
The whole‑genome techniques include the restriction
endonuclease analysis (REA) typing, the pulsed‑field
gel electrophoresis, the RFLP, WGS, and whole‑genome
microarray. REA typing has helped in tracing the source of
infections that resulted from within‑herd transmission and
movement of livestock, thus allowing for outbreak source
differentiation among wildlife reservoirs in different regions
or reinfection from herd movement.[57‑59]
Other techniques termed the partial genome techniques with
specific repeated sequences include the tandem‑repeated
sequences and non‑tandem repeated sequences; random
sequences as the random amplified polymorphic
deoxyribonucleic acid analysis; rarely used now due to
its lack of reproducibility and poor discrimination. Others
including house‑keeping genes as the multilocus sequence
typing, typing, and regions of difference and single nucleotide
polymorphism (SNP).[60] SNP typing has a clear discriminatory
Biomedical and Biotechnology Research Journal ¦ Volume 3 ¦ Issue 4 ¦ October-December 2019
power though there is a need to test a large set of genes to
achieve a satisfactory resolution. It has been used to show some
phylogenetic distribution.[60,61] It is not used too commonly
because it is a two‑step technique which makes it expensive and
time‑consuming.[62] SNP‑based analyses have demonstrated the
genotypic differences within M. bovis strains and differentiated
these strains from M. tuberculosis strains representing diversity
in time and space. It has equally provided population genetic
frameworks that may aid in identifying factors responsible for
the wide host range and disease phenotypes of M. bovis.[63]
Although the various molecular epidemiological studies
enable the traceability of M. bovis infections and help in the
detection of the outbreaks source,[64] M. bovis genotyping
systems still need to be standardized.[65] VNTR typing (that
provides better discrimination than all other methods except
for REA),[55] and spoligotyping, either alone or together,
have now become the preferred approaches as they are
robust and amenable to electronic analysis and comparison.
Spoligotyping though only moderately discriminatory can
be easily applied to large numbers of isolates. It is the most
common epidemiological molecular‑typing method applied to
strains of M. bovis[66] [Table 1].
Molecular studies of Mycobacterium bovis infection in
human in Africa
Spoligotyping and VNTR have aided the survey of
isolates,[80] and clearly illustrated the geospatial localization
of epidemiologically distinct group of M. bovis types across
wildlife populations and cattle. Although the discriminatory
power of the different typing techniques for M. bovis varies
by country, M. bovis genotypes of human origin (with rare
exception of M. bovis recovered from humans not matching
the molecular type of the cattle strains)[20] generally parallel
those of livestock within the same country, which supports
the presence of an epidemiological link between human
and animal M. bovis TB cases.[30,81] Three clonal lineages of
M. bovis: African 1, African 2, and European 1, with a distinct
spoligotype signature, have been characterized with the African
1 and African 2 groups shown to be restricted to Central‑west
Africa and East Africa, respectively, while the European 1
members being globally distributed.[82]
The African 1 (Af1) clonal complex of M. bovis defined by
a specific chromosomal deletion  (RDAf1) and the absence
of spacer 30 in the standard spoligotype has been shown to
be present at high frequency in cattle samples from several
sub‑Saharan west/central African countries such as Mali,
Cameroon, Nigeria, and Chad, suggesting that the recent
mixing of strains between countries is not common in this
area of Africa.[66] However, two groups of M. bovis have been
shown to circulate in Burkina Faso: a major group (Af1) and a
minor group belonging to the putative Af5 clonal complex.[15]
The Af1‑specific deletion has not been identified in M. bovis
isolates in several other African countries such as Algeria,
Burundi, Ethiopia, Madagascar, Mozambique, South Africa,
Tanzania, and Uganda, as well as from Europe and South
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Table 1: Mycobacterium bovis prevalence and typing techniques from livestock and human in Africa
Country Sample source Prevalence  Typing methods Predominent livestock/human spoligotypes References
Algeria Cattle 1.21% Spoligotyping, VNTR SB0120, SB0121, SB0134, SB0941 [67]
Burkina Faso Cattle 25.75% Spoligotyping, RDAf1 SB0944 [15]
Human 1.22% deletion, MIRU‑VNTR
Ethiopia Cattle 11.1% Spoligotyping SB1176 (41.2%) [68]
SB0133 (14.7%)
Cattle LSP, Spoligotyping SB0133, SB0933, [40]
Camels/goats 11.7% SB1942, SB1983
Human 1.9% SB0133, SB0303
Ghana Human 0.6% Spoligotyping, LSP, IS6110, SB0136 [69]
Madagascar Cattle/goat and 162 animals rpoß PCR [70]
humans 18 humans RFLP, Spoligotyping
(1994-2000)
Mali Cattle 33.33% Spoligotyping SB0300, SB0134 [71]
Morocco Cattle 1.7% Spoligotyping, SB0121, SB0265, SB0120, SB0125 SB0869 [72]
deletion typing
Nigeria Cattle 23.5% Spoligotyping SB0944, SB0952 [20]
Human 5% LSP, VNTR
South Africa Wild animals 31 Mycobacterium Spoligotyping, VNTR, SB0130 [73]
bovis isolates PGRS, IS6110-RFLP
Cattle Spoligotyping, VNTR, SB0131 [74]
IS6110-RFLP, PGRS SB0130
SB0140
Tanzania Cattle, wildlife 0.83% Spoligotyping, SB0133 [75]
Human 0% MIRU‑VNTR
Cattle 8.9% Spoligotype, RD4 and RD9 SB1467 [76]
PCR assays
Tunisia Raw milk 4.9% Spoligotyping, VNTR SB1093, SB 1003 [77]
Zambia Cattle 4.46% Spoligotyping SB0120 [78]
Human 0.86% SB0120 [79]
MIRU ‑ VNTR: Mycobacterial Interspersed Repetitive Units ‑ Variable Number of Tandem Repeats, RDAf1: Af1‑specific deletion, LSP: Large Sequence
Polymorphism, RFLP: Restriction fragment length polymorphism, PGRS: Polymorphic G–C‑rich sequences

America pointing out its geographic localization to several Strategies to Improve Bovine Tuberculosis
African countries.[66]
Control in Africa
Molecular studies have shown that the isolation rate
of M. bovis from human patients is highly variable in
Roadmap for zoonotic tuberculosis and one heath strategy
The strategies required for the control and elimination of BTB
Africa, [20,83‑86] and despite limited human‑to‑human
are defined, though several challenges including among others
transmission of zoonotic TB, the infection often causes
financial constraints, scarcity of trained professionals, lack of
extrapulmonary disease in humans,[87] with severe effects on
political will, and the underestimation of the importance of
livelihoods.[53] Higher prevalence (up to 30%) of M. bovis
zoonotic TB in both the animal and public health sectors by national
is usually recorded in rural settings where the population is
governments and donor agencies hinder the applications of the
more closely associated with livestock compared to urban
control measures, especially in Africa.[87] There is an initiative
areas.[88] Some studies have shown that the mutations on the
to combat BTB in humans, alongside other zoonotic diseases in
PhoPR regulation system in the MTBC lead to a decreased
an integrated approach to improve human and animal health.[95]
affinity of M. bovis to humans.[89,90] In Africa, M. bovis
A multi‑sectorial collaboration pioneered by the World Health
bacillus Calmette–Guérin (BCG) vaccination generalized
Organization (WHO), OIE, and FAO under a tripartite partnership
in several countries is hypothesized to have lead in M. bovis
and providing guidance on how to reduce zoonoses risks at the
strain suppression with the emergence and dissemination of
human‑animal‑environment interface has been developed to
a different lineage.[91] BCG vaccination has been implicated
jointly pursue the One Health approach and working in close
in the decrease of M. africanum which is less virulent in
partnership with research institutions, academia, intergovernmental
experimental animal models than M. tuberculosis,[92] and
organizations, the private sector, nongovernmental organizations,
equally its possible role in the selection of resistant strains
civil societies, and all stakeholders.[96]
with BCG‑induced immunity has been suggested to explain
the expansion of Beijing family strains and the predominance The roadmap calling for concerted action through broad
of other families in certain geographic settings.[93,94] engagement across political, financial, and technical levels lays

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Pokam, et al.: BTB control in Africa
down ten priorities grouped into three core themes (improve
the scientific evidence base, reduce transmission at the
animal‑human interface, strengthen intersectoral, and
collaborative approaches) with milestones defined for the
short‑term, by 2020, and medium‑term, by 2025, in a WHO’s
End TB Strategy by 2030.[97] This initiative, however, will
require a strong political will (still absent in Africa) to address
the basic utilities and focus first on appropriate human and
infrastructure capacities building to ensure its applications. It
has to take into account the economic, cultural, technological,
and logistical issues encountered in Africa. The success of
the “One Health” approach has shown the potential benefit of
human and veterinary medical health professionals working
cooperatively to identify “shared risks” to humans and
animals.[98] This synergism will advance health care, improve
biomedical research discoveries, and enhance public health
efficacy (http://www.onehealthinitiative.com/about.php).
Surveillance, capacity building, and innovation strategies
The complex nature of zoonotic diseases and the limited
resources is a reminder of the importance of disease
surveillance in the most affected countries.[99] The human
burden of disease cannot be reduced without improving
standards of food safety and controlling bovine TB in
the animal reservoir.[97] Low prevalence of BTB has been
attributed to effective surveillance activities which included
both active surveillance with detection during meat inspection,
targeted screening of cattle, laboratory testing, restriction
of movement from infected herds, and awareness creation
on its economic and health implications, as well as passive
surveillance with reporting from dairy farms and cattle
herdsmen to veterinary staff. BTB surveillance in Africa is
either weak or nonexistent and will require an enforcement of
known regulations which has shown to lead to a reduction of
human TB infections due to M. bovis.[100] Building capacity for
surveillance of BTB will include building technical capacity
for quality control of tuberculin and tuberculin testing and
designing epidemiological surveys; strengthening capacity for
epidemiological data analysis; improving the capacity of meat
inspectors to recognize and to report TB on carcasses; building
and strengthening existing capacities of laboratory diagnosis
of animal TB; and building leadership of veterinarians and
farmer/nomadic population awareness, trust, and advocacy.[101]
Emphasis has to be laid on the capacity building for physicians,
veterinarians, scientists, and technicians trained for an
adequate surveillance system, including a strong laboratory
network, which is a key component of a meaningful prevention
and control of BTB diseases. The development of capable
diagnostic facilities is paramount for dealing with BTB
surveillance for differentiation from other mycobacteria for
effective treatment and control. Although some progress
has been made, there is a need for continued investment
and political commitment to meet the enormous persisting
challenge of BTB in Africa.[99]
The increasing use of bioinformatics tools and linkage of
epidemiological and molecular surveillance data on animal and
Biomedical and Biotechnology Research Journal ¦ Volume 3 ¦ Issue 4 ¦ October-December 2019
human health to understand better the evolution of pathogens
crossing between animal and human populations, and the
factors driving pathogen adaptation, as well as the introduction
of routine molecular information within the African public
health and veterinary systems coupled with proper capacity
building will help to improve zoonotic TB surveillance. The
progress on genomic sequencing of nonhuman animals will
open up opportunities for comparative genomic approaches to
understanding differential susceptibility between species, and
the genomic development in Africa could result in significant
progress to monitor and improve the environmental conditions
that are critical to both human and animal health. The molecular
and biodiversity information introduced into public health
practice in Africa will enable epidemiologists to target their
population‑level surveillance of zoonotic TB to the most
susceptible at‑risk animals.[98]
The linkages between the types of data streams to identify key
environmental factors driving disease emergence in animal
and human populations can be established and analyzed
using the on‑time and real‑time strengthening communication
and awareness creation of FAO established systems as the
Global Early Warning System for transboundary animal
diseases and major zoonoses.[102] Biosurveillance system such
as the real‑time outbreak and disease surveillance system
at the Automated Epidemiologic Geotemporal Integrated
Surveillance could be modified to support the linkage of
animal and human data for monitoring of zoonotic diseases
in Africa.[103,104]
Mycobacterium bovis vaccination as control strategy in
Africa
A long‑lasting solution such as the protection of cattle against
BTB by vaccination could be an important control strategy in
countries where there is the persistence of M. bovis infection in
wildlife and in developing countries where it is not economical
to implement a “test and slaughter” control program.[105]
Vaccination of cattle represents an alternative intervention
strategy to reduce the impact of BTB on livestock productivity
and human health in developing countries. The only currently
available vaccine against TB is the human vaccine ‑ M. bovis
BCG. Although early field trials with BCG M. bovis vaccine
in cattle have been shown to produce disappointing results,
with the induction of tuberculin skin‑test reactivity following
vaccination and low levels of protection,[105] an encouraging
protective effect of BCG against BTB in a natural transmission
setting in Africa has been demonstrated.[106] However, a
large number of experiments and trials have shown variable
efficacies,[107] and the variation in the level of protection is
explained with the increased in the understanding of immunity
to TB.[108] This includes the use of different BCG strains, very
high doses of BCG administered, pre‑existing exposure to
environmental mycobacteria, lack of long‑term protection,
very high levels of natural challenge, and M. bovis exposure
from milk of infected cows prior to vaccination. This variable
levels of protection observed in field trials mirror the situation
in humans where BCG vaccine efficacy against pulmonary
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Pokam, et al.: BTB control in Africa
TB in children and adults is highly variable.[109] Moreover,
a complementary diagnostic test to differentiate between
vaccinated animals and those infected with M. bovis is required
so that test‑ and slaughter‑based control strategies can continue
alongside vaccination.[110]
Peptide and protein cocktails has been suggested to
discriminate between M. bovis infection and BCG vaccination
in animals, [110] and a prototype differential diagnosis of
infected and vaccinated animals reagents based on the early
secreted antigen target 6 kDa protein and culture filtrate
protein 10 present in M. bovis but absent from BCG in the
bovine interferon‑gamma (Bovigam) test assay has been
shown to be able to distinguish between vaccinated animals
that were protected against BTB and those animals that
were not in Africa.[106] In the past two decades, considerable
progress has been made in the development and evaluation
of TB vaccines for cattle, with new attenuated mycobacterial
vaccines providing an alternative to the use of BCG vaccine
and subunit vaccines to boost protection induced by BCG.
With the development of tests to differentiate infected from
vaccinated animals, it is now feasible to use vaccines to assist
in the control of this disease.[108]
Conclusion
Understanding the epidemiology of the infection within and
between species is crucial to the control of BTB in both domestic
and wild animals. The transmission is highest from infected
domestic animals to susceptible wildlife (and vice versa) when
they share pasture or territory.[111] BTB prevention and control
need to be step up in the African region and will require a
constant effort to be eradicated. As the trade of animal products,
the urbanization and the movements of people intensify; the risk
of its introduction/reintroduction in different areas is challenging
and thus will require a country level as well as regional or
subregional surveillance scheme to tackle its spread. Improving
laboratory capacities for rapid and appropriate diagnosis of strain
involved in human transmission will improve treatment and limit
haphazard management of acid‑fast bacilli in patients with the
direct consequence of increased drug resistance. The evidence
of genomic overlap of M. bovis between cattle and man points
out the need for synergy of veterinary and medical policies in
the control of TB for its eradication.
Acknowledgment
The authors are grateful to Dr. Afutendem Lucas of the
University of Dschang (English Language ‑ Applied
Linguistics, Phonology) for English language editing and
proofreading.
Financial support and sponsorship
This work was supported by the Bill and Melinda Gates
Foundation to B.D.T.P, under the Postdoctoral and Postgraduate
Training in Infectious Diseases Research awarded to the
Noguchi Memorial Institute for Medical Research (Global
Health Grant number OPP52155). The funding body has no
role in the design and writing of the manuscript.
224 Biomedical and Biotechnology Research Journal ¦ Volume 3 ¦ Issue 4 ¦ October-December 2019
Conflicts of interest
There are no conflicts of interest.
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