The predominant spoligotype, widely dispersed geographically (Table 1 &2), was found in the international data base to have a pattern with a spoligotype number SB0120 with the corresponding hexacode of 6F-5F-5F-7F-FF-60. Five out of the six study districts had this predominant spoligotype, and Namwala district accounted for 30% of spoligotype SB0120. The second most predominant spoligotype had a pattern named SB0871 with a corresponding hexacode of 6F-4F-5F-7F-FF-60. Isolates C14 was

named SB1572 with a hexacode number of 6F-5F-5F-7F-FF-40, isolate C16 was CB-839 mouse SB1536 with a hexacode number of 2F-5F-5F-6F-FF-60 and isolate C19 was SB0162 with a hexacode number of 00-00-00-0F-FF-60. The distribution

of these spoligotypes on the international data base is shown in Table 2. Table 2 Major Spoligotypes in Zambia find more Spoligotype1 Shared type2 Geographical distribution Sp1 SB0120 France, Belgium, Brazil, South Africa, Sri Lanka, Iran, The Netherlands, Spain, China, Japan, Portugal, Russia, Denmark, Zambia Sp2 SB0871 France Sp3 SB1763* Zambia Sp4 SB1764* Zambia Sp5 SB1572 Italy Sp6 SHP099 solubility dmso SB1765* Zambia Sp7 SB1536 Italy Sp8 SB1766* Zambia Sp9 SB1767* Zambia Sp10 SB0162 Belgium 1 Arbitrary spoligotype designation 2 Shared type, designation of the spoligotype in the World Spoligotype Database. *New Spoligotype assigned by http://​www.​mbovis.​org Five individually occurring isolates (16.1%) displayed new spoligo patterns that have not yet been described on the international PIK-5 spoligotyping data base (Figure 2 and Table 2). These isolates

originated from Namwala district (isolate C26, 42 and C41); from Mumbwa (isolate C21); and from Monze (isolate C9) (Table 1 and Figure 2). These new patterns were allotted new spoligo numbers as SB1763 (hex code 66-03-5F-6D-FF-60), SB1764 (hex code 60-0F-1F-6C-FF-00), SB1765 (hex code 2F-5F-5F-7F-FF-40), SB1766 (hex code 6F-4F-1F-6F-FF-60) and SB1767 (hex code 62-0E-50-09-FF-40) by http://​www.​mbovis.​org Table 2. The technique showed a good discrimination power; Hunter Gaston Discriminatory Index (HGDI = 0.98) (Table 1 and Figure 2.). Discussion Our results do not agree with what has been found in other parts of Africa [21, 22], where more than 40% of the animals with tuberculous lesions had Non-tuberculous Mycobacteria (NTM). In this study, only two animals had mycobacteria other than M. bovis. However, our findings tie up with a similar study conducted in Algeria [23]. Whereas excluding the differences in bacterial species as accounting for these observations [23], strain isolation has been found to be dependant on the specific type of media used [24]. The usage of specific culture media such as Stonebrink has been shown to increase the recovery and discrimination of strains on culture [25, 26].