64 R.A. Skuce et al.
Longitudinal typing data were observed to track
reduction in pathogen diversity as France’s erad-
ication scheme gathered momentum in the pre-
ceding decade (Hauer et al., 2015). MLVA has
been applied to cattle M. bovis isolates in order to
better understand the population structure and
epidemiology of the pathogen in Belgium (Allix
et al., 2006). Human and deer isolates from Swe-
den have been genotyped using IS 6110 RFLP
(Szewzyk et al., 1995). The genetic differences
observed confirmed that both hosts were affected
by distinct bacterial lineages, thereby ruling out
disease transmission (Szewzyk et al., 1995).
5.5.2 AfricaSpoligotype and MLVA analysis of M. bovis iso-
lates from cattle slaughtered in Algeria con-
firmed the presence of molecular types
associated with previously documented Euro-
pean strains (Sahraoui et al., 2009). In Tunisia,
initial development of spoligotyping and an
MLVA panel has taken place and revealed the
widespread distribution of molecular types asso-
ciated with European cattle (Lamine-Khemiri
et al., 2013). Both studies presented data consis-
tent with the legacy of European colonialism in
North Africa. Initial efforts to characterize the
M. bovis population of Cameroon relied on RFLP
and pulsed field gel electrophoresis (PFGE) meth-
ods (Njanpop-Lafourcade et al., 2001). Subse-
quent application of spoligotyping and MLVA to
180 Cameroonian cattle samples detected
greater diversity and molecular types associated
with neighbouring Nigeria (Cadmus et al.,
2011). More recently, comparative genomics
and genotyping of a large sequence polymor-
phism has enabled the detection of a dominant
M. bovis clonal complex, named Af1, in Saharan
West Africa encompassing the cattle popula-
tions of Mali, Nigeria, Chad and Cameroon
(Müller et al., 2009). MLVA of cattle isolates
from all territories indicated very limited sharing
of molecular types indicating recent mixing via
cattle movement (Müller et al., 2009). Similar
work in neighbouring Burkina Faso revealed the
same clonal complex dominance and sharing of
spoligotypes, with MLVA, suggesting country-
specific evolution of molecular types (Sanou
et al., 2014).
In Ethiopia early application of MLVA and
spoligotyping revealed novel molecular types in
the Addis Ababa area (Ameni et al., 2007). Fur-
ther application of spoligotyping in northern
Ethiopia revealed clustering of six molecular
types, three of which were novel (Ameni et al.,
2010). MLVA and spoligotyping have been used
in central Ethiopia (Firdessa et al., 2012), while
further optimization of the techniques has been
published (Biffa et al., 2014). In Uganda, spoligo-
typing and RFLP typing confirmed M. bovis
infection and distinguished members of the MTC
(Oloya et al., 2007). In Tanzania, MLVA and
spoligotyping have been applied to M. bovis iso-
lates from multiple hosts, including humans,
livestock and wildlife, where ongoing transmis-
sion was occurring, but transmission to humans
was not occurring (Katale et al., 2015). Com-
parative genomics revealed a chromosomal dele-
tion/large sequence polymorphism unique to
M. bovis isolates from the East African countries
of Ethiopia, Uganda, Burundi and Tanzania,
marking a further clonal complex named as Af2
(Berg et al., 2012).
In South Africa, IS 6110 RFLP was used to
investigate the epidemiology of M. bovis trans-
mission between buffalo and other wildlife spe-
cies in the Kruger National Park (Michel et al.,
2009). Molecular typing confirmed that buffalo
interacting with cattle was the source of M. bovis
entering the park (Michel et al., 2009). Local
optimization of an MLVA panel for the Kruger
National Park facilitated distinction of related
and unrelated cases (Hlokwe et al., 2013), and
subsequently MLVA and spoligotyping were used
to detect spillover from wildlife hosts in the
greater Kruger National Park area to bordering
regions with livestock (Musoke et al., 2015).
Further north in Zambia, spoligotyping has
revealed shared molecular types of M. bovis in
cattle and humans affected by tuberculosis sug-
gesting zoonotic transmission (Malama et al.,
2014).5.5.3 The AmericasIn the USA, RFLP methods were applied initially
to isolates from cattle, captive elk and other wild-
life species (Whipple et al., 1997). In general, the
same M. bovis molecular type was observed for