NRU :: Browsing by Author "Siegismund, Hans R."

Browsing by Author "Siegismund, Hans R."

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Characterisation of recent foot-and-mouth disease viruses from African buffalo (Syncerus caffer) and cattle in Kenya is consistent with independent virus populations
(BMC veterinary research, 2015) Wekesa, Sabenzia N.; Sangula, Abraham K.; Belsham, Graham J.; Tjørnehøj, Kirsten; Muwanika, Vincent B.; Gakuya, Francis; Mijele, Dominic; Siegismund, Hans R.
Understanding the epidemiology of foot-and-mouth disease (FMD), including roles played by different hosts, is essential for improving disease control. The African buffalo (Syncerus caffer) is a reservoir for the SAT serotypes of FMD virus (FMDV). Large buffalo populations commonly intermingle with livestock in Kenya, yet earlier studies have focused on FMD in the domestic livestock, hence the contribution of buffalo to disease in livestock is largely unknown. This study analysed 47 epithelia collected from FMD outbreaks in Kenyan cattle between 2008 and 2012, and 102 probang and serum samples collected from buffalo in three different Kenyan ecosystems; Maasai-Mara (MME) (n = 40), Tsavo (TSE) (n = 33), and Meru (ME) (n = 29). Results: Antibodies against FMDV non-structural proteins were found in 65 of 102 (64%) sera from buffalo with 44/102 and 53/102 also having neutralising antibodies directed against FMDV SAT 1 and SAT 2, respectively. FMDV RNA was detected in 42% of the buffalo probang samples by RT-qPCR (Cycle Threshold (Ct) ≤32). Two buffalo probang samples were positive by VI and were identified as FMDV SAT 1 and SAT 2 by Ag-ELISA, while the latter assay detected serotypes O (1), A (20), SAT 1 (7) and SAT 2 (19) in the 47 cattle epithelia. VP1 coding sequences were generated for two buffalo and 21 cattle samples. Phylogenetic analyses revealed SAT 1 and SAT 2 virus lineages within buffalo that were distinct from those detected in cattle. Conclusions: We found that FMDV serotypes O, A, SAT 1 and SAT 2 were circulating among cattle in Kenya and cause disease, but only SAT 1 and SAT 2 viruses were successfully isolated from clinically normal buffalo. The buffalo isolates were genetically distinct from isolates obtained from cattle. Control efforts should focus primarily on reducing FMDV circulation among livestock and limiting interaction with buffalo. Comprehensive studies incorporating additional buffalo viruses are recommended.
Characterization of Foot-And-Mouth Disease Viruses (FMDVs) from Ugandan Cattle Outbreaks during 2012-2013: Evidence for Circulation of Multiple Serotypes
(PLoS One, 2015) Namatovu, Alice; Tjørnehøj, Kirsten; Belsham, Graham J.; Dhikusooka, Moses T.; Wekesa, Sabenzia N.; Muwanika, Vincent B.; Siegismund, Hans R.; Ayebazibwe, Chrisostom
To investigate the foot-and-mouth disease virus (FMDV) serotypes circulating in Uganda’s cattle population, both serological and virological analyses of samples from outbreaks that occurred during 2012–2013 were performed. Altogether, 79 sera and 60 oropharyngeal fluid (OP)/ tissue/oral swab samples were collected from herds with reported FMD outbreaks in seven different Ugandan districts. Overall, 61/79 (77%) of the cattle sera were positive for antibodies against FMDV by PrioCHECK FMDV NS ELISA and solid phase blocking ELISA detected titres 80 for serotypes O, SAT 1, SAT 2 and SAT 3 in 41, 45, 30 and 45 of these 61 seropositive samples, respectively. Virus neutralisation tests detected the highest levels of neutralising antibodies (titres 45) against serotype O in the herds from Kween and Rakai districts, against SAT 1 in the herd from Nwoya district and against SAT 2 in the herds fromKiruhura, Isingiro and Ntungamo districts. The isolation of a SAT 2 FMDV from Isingiro was consistent with the detection of high levels of neutralising antibodies against SAT 2; sequencing (for the VP1 coding region) indicated that this virus belonged to lineage I within this serotype, like the currently used vaccine strain. From theWakiso district 11 tissue/swab samples were collected; serotype A FMDV, genotype Africa (G-I), was isolated from the epithelial samples. This study shows that within a period of less than one year, FMD outbreaks in Uganda were caused by four different serotypes namely O, A, SAT 1 and SAT 2. Therefore, to enhance the control of FMD in Uganda, there is need for efficient and timely determination of outbreak virus strains/serotypes and vaccine matching. The value of incorporating serotype A antigen into the imported vaccines along with the current serotype O, SAT 1 and SAT 2 strains should be considered.
Diversity and transboundary mobility of serotype O foot-and-mouth disease virus in East Africa: Implications for vaccination policies
(Infection, Genetics and Evolution, 2010) Balinda, Sheila N.; Sangula, Abraham K.; Heller, Rasmus; Muwanika, Vincent B.; Belsham, Graham J.; Masembe, Charles; Siegismund, Hans R.
Foot-and-mouth disease (FMD) virus serotype O has been responsible for most reported outbreaks of the disease in East Africa. A sustained campaign for the past 40 years to control FMD mainly by vaccination, combined with quarantine and zoosanitary measures has been undertaken with limited success. We investigated the genetic relationships among serotype O strains in eastern Africa using complete VP1 coding region sequences obtained from 46 FMD virus isolates collected in Kenya in the years 1964–2008 and 8 Ugandan isolates collected between 1999 and 2006. In addition, 21 selected FMDV sequences from Genbank representing reference strains from eastern Africa and elsewhere were included in the Bayesian inference analyses and the detection of selection forces. The results confirmed previous observations that eastern Africa harbours four distinct topotypes (clades with >15% sequence divergence). All but one strain isolated post-2000 belonged to topotypes EA-2, EA-3 and EA-4, while all three vaccines have been based on strains in the EA-1 topotype. The estimated dN/dS ratios across the individual codons of the entire VP1 coding region revealed that purifying (negative) selection constituted the dominant evolutionary force. Cross-border disease transmission within the region has been suggested with probable incursions of topotypes EA-3 and EA-4 into Kenya and Uganda from neighboring Ethiopia and Sudan. Weconclude that the vaccines have probably been effective in controlling EA-1, but less so for the other topotypes and propose a more comprehensive representation of topotypes in the development of new vaccines in recognition of the considerable diversity and transboundary nature of serotype O.
Evolutionary analysis of foot-and-mouth disease virus serotype SAT 1 isolates from east Africa suggests two independent introductions from southern Africa
(BMC evolutionary biology,, 2010) Sangula, Abraham K.; Belsham, Graham J.; Muwanika, Vincent B.; Heller, Rasmus; Balinda, Sheila N.; Masembe, Charles; Siegismund, Hans R.
In East Africa, foot-and-mouth disease virus serotype SAT 1 is responsible for occasional severe outbreaks in livestock and is known to be maintained within the buffalo populations. Little is known about the evolutionary forces underlying its epidemiology in the region. To enhance our appreciation of the epidemiological status of serotype SAT 1 virus in the region, we inferred its evolutionary and phylogeographic history by means of genealogy-based coalescent methods using 53 VP1 coding sequences covering a sampling period from 1948-2007. Results: The VP1 coding sequence of 11 serotype SAT 1 FMD viruses from East Africa has been determined and compared with known sequences derived from other SAT 1 viruses from sub-Saharan Africa. Purifying (negative) selection and low substitution rates characterized the SAT 1 virus isolates in East Africa. Two virus groups with probable independent introductions from southern Africa were identified from a maximum clade credibility tree. One group was exclusive to Uganda while the other was present within Kenya and Tanzania. Conclusions: Our results provide a baseline characterization of the inter-regional spread of SAT 1 in sub-Saharan Africa and highlight the importance of a regional approach to trans-boundary animal disease control in order to monitor circulating strains and apply appropriate vaccines.
Genetic consequences of population expansions and contractions in the common hippopotamus (Hippopotamus amphibius) since the Late Pleistocene
(Molecular Ecology, 2015) Stoffel, Céline; Dufresnes, Christophe; Okello, John Bosco A.; Noirard, Christian; Joly, Pierre; Nyakaana, Silvester; Muwanika, Vincent B.; Alcala, Nicolas; Vuilleumier, Séverine; Siegismund, Hans R.; Fumagalli, Luca
Over the past two decades, an increasing amount of phylogeographic work has substantially improved our understanding of African biogeography, in particular the role played by Pleistocene pluvial-drought cycles on terrestrial vertebrates. However, still little is known on the evolutionary history of semi-aquatic animals, which faced tremendous challenges imposed by unpredictable availability of water resources. In this study, we investigate the Late Pleistocene history of the common hippopotamus (Hippopotamus amphibius), using mitochondrial and nuclear DNA sequence variation and range-wide sampling. We documented a global demographic and spatial expansion approximately 0.1-0.3 My ago, most likely associated with an episode of massive drainage overflow. These events presumably enabled a historical continent-wide gene flow among hippopotamus populations, and hence no clear continental-scale genetic structuring remains. Nevertheless, present-day hippopotamus populations are genetically disconnected, probably as a result of the mid- Holocene aridification and contemporary anthropogenic pressures. This unique pattern contrasts with the biogeographic paradigms established for savannah-adapted ungulate mammals and should be further investigated in other water-associated taxa. Our study has important consequences for the conservation of the hippo, an emblematic but threatened species that requires specific protection to curtail its long-term decline.
Laboratory capacity for diagnosis of foot-and-mouth disease in Eastern Africa: implications for the progressive control pathway
(BMC veterinary research, 2013) Namatovu, Alice; Wekesa, Sabenzia N.; Tjørnehøj, Kirsten; Dhikusooka, Moses T.; Muwanika, Vincent B.; Siegismund, Hans R.; Ayebazibwe, Chrisostom
Accurate diagnosis is pertinent to any disease control programme. If Eastern Africa is to work towards control of foot-and-mouth disease (FMD) using the Progressive Control Pathway for FMD (PCP-FMD) as a tool, then the capacity of national reference laboratories (NRLs) mandated to diagnose FMD should match this task. This study assessed the laboratory capacity of 14 NRLs of the Eastern Africa Region Laboratory Network member countries using a semi-structured questionnaire and retrospective data from the World Reference Laboratory for FMD annual reports and GenbankW through National Centre for Biotechnology Information for the period 2006–2010. Results: The questionnaire response rate was 13/14 (93%). Twelve out of the 13 countries/regions had experienced at least one outbreak in the relevant five year period. Only two countries (Ethiopia and Kenya) had laboratories at biosecurity level 3 and only three (Ethiopia, Kenya and Sudan) had identified FMD virus serotypes for all reported outbreaks. Based on their own country/region assessment, 12/13 of these countries /regions were below stage 3 of the PCP-FMD. Quarantine (77%) and vaccination (54%) were the major FMD control strategies employed. The majority (12/13) of the NRLs used serological techniques to diagnose FMD, seven used antigen ELISA and three of these (25%) also used molecular techniques which were the tests most frequently requested from collaborating laboratories by the majority (69%) of the NRLs. Only 4/13 (31%) participated in proficiency testing for FMD. Four (31%) laboratories had no quality management systems (QMS) in place and where QMS existed it was still deficient, thus, none of the laboratories had achieved accreditation for FMD diagnosis. Conclusions: This study indicates that FMD diagnostic capacity in Eastern Africa is still inadequate and largely depends on antigen and antibody ELISAs techniques undertaken by the NRLs. Hence, for the region to progress on the PCP-FMD, there is need to: implement regional control measures, improve the serological diagnostic test performance and laboratory capacity of the NRLs (including training of personnel as well as upgrading of equipment and methods, especially strengthening the molecular diagnostic capacity), and to establish a regional reference laboratory to enforce QMS and characterization of FMD virus containing samples.
Molecular characterization of SAT 2 foot-and-mouth disease virus from post-outbreak slaughtered animals: implications for disease control in Uganda
(Epidemiology & Infection, 2010) Balinda, Sabenzia N.; Belsham, Graham J.; Masembe, Charles; Sangula, Abraham K.; Siegismund, Hans R.; Muwanika, Vincent B.
In Uganda, limiting the extent of foot-and-mouth disease (FMD) spread during outbreaks involves short-term measures such as ring vaccination and restrictions of the movement of livestock and their products to and from the affected areas. In this study, the presence of FMD virus RNA was investigated in cattle samples 3 months after FMD quarantine measures had been lifted following an outbreak in 2004. Oropharyngeal tissue samples were obtained from 12 cattle slaughtered in a small town abattoir in Kiboga. FMD virus RNA was detected by diagnostic RT–PCR in nine of the 12 tissue samples. Part of the coding region for the capsid protein VP1 was amplified and sequenced. All samples were identified as belonging to the SAT 2 serotype. The implications for FMD control of both virus introduction into Uganda and the presence of carrier animals following outbreaks are discussed.
Patterns, risk factors and characteristics of reported and perceived foot-and-mouth disease (FMD) in Uganda
(Tropical Animal Health and Production, 2010) Ayebazibwe, Chrisostom; Tjørnehøj, Kirsten; Mwiine, Frank N.; Muwanika, Vincent B.; Okurut, Anna Rose Ademun; Siegismund, Hans R.; Alexandersen, Soren
Patterns of outbreaks of foot-and-mouth disease (FMD) in Uganda were elucidated from spatial and temporal retrospective data retrieved from monthly reports from District Veterinary Officers (DVOs) to the central administration for the years spanning 2001–2008. An assessment of perceived FMD occurrence, risk factors and the associated characteristics was made based on semi-structured questionnaires administered to the DVOs. During this period, a total of 311 FMD outbreaks were reported in 56 (70%) out of Uganda’s 80 districts. The number of reported FMD outbreaks changed over time and by geographical regions. Occurrence of FMD was significantly associated with the dry season months (p = 0.0346), the time when animals movements are more frequent. The average number of FMD outbreaks was higher for some sub-counties adjacent to national parks than for other sub-counties, whilst proximity to international border only seemed to play a role at the southern border. DVOs believed that the major risk factor for FMD outbreaks was animal movements (odds ratio OR 50.8, confidence interval CI 17.8–144.6) and that most outbreaks were caused by introduction of sick animals.
Patterns, Risk Factors And Characteristics Of Reported And Perceived Foot-And-Mouth Disease (FMD) In Uganda
(Tropical Animal Health and Production, 2010) Ayebazibwe, Chrisostom; Tjørnehøj, Kirsten; Mwiine, Frank N.; Muwanika, Vincent B.; Okurut, Anna Rose Ademun; Siegismund, Hans R.; Alexandersen, Soren
Patterns of outbreaks of foot-and-mouth disease (FMD) in Uganda were elucidated from spatial and temporal retrospective data retrieved from monthly reports from District Veterinary Officers (DVOs) to the central administration for the years spanning 2001–2008. An assessment of perceived FMD occurrence, risk factors and the associated characteristics was made based on semi-structured questionnaires administered to the DVOs. During this period, a total of 311 FMD outbreaks were reported in 56 (70%) out of Uganda’s 80 districts. The number of reported FMD outbreaks changed over time and by geographical regions. Occurrence of FMD was significantly associated with the dry season months (p = 0.0346), the time when animals movements are more frequent. The average number of FMD outbreaks was higher for some sub-counties adjacent to national parks than for other sub-counties, whilst proximity to international border only seemed to play a role at the southern border. DVOs believed that the major risk factor for FMD outbreaks was animal movements (odds ratio OR 50.8, confidence interval CI 17.8–144.6) and that most outbreaks were caused by introduction of sick animals.
Population Genetic Structure of Savannah Elephants in Kenya: Conservation and Management Implications
(Journal of heredity, 2008) Okello, John B. A.; Masembe, Charles; Rasmussen, Henrik B.; Wittemyer, George; Omondi, Patrick; Kahindi, Onesmas; Muwanika, Vincent B.; Arctander, Peter; Douglas-Hamilton, Iain; Nyakaana, Silvester; Siegismund, Hans R.
We investigated population genetic structure and regional differentiation among African savannah elephants in Kenya using mitochondrial and microsatellite markers. We observed mitochondrial DNA (mtDNA) nucleotide diversity of 1.68% and microsatellite variation in terms of average number of alleles, expected and observed heterozygosities in the total study population of 10.20, 0.75, and 0.69, respectively. Hierarchical analysis of molecular variance of mtDNA variation revealed significant differentiation among the 3 geographical regions studied (FCT 5 0.264; P , 0.05) and a relatively lower differentiation among populations within regions (FSC 5 0.218; P , 0.0001). Microsatellite variation significantly differentiated among populations within regions (FSC 5 0.019; P , 0.0001) but not at the regional levels (FCT 5 0.000; P . 0.500). We attribute the high differentiation at the mitochondrial genome to the matrilineal social structure of elephant populations, female natal philopatry, and probably ancient vicariance. Lack of significant regional differentiation at the nuclear loci vis-a-vis strong differences at mtDNA loci between regions is likely the effect of subsequent homogenization through male-mediated gene flow. Our results depicting 3 broad regional mtDNA groups and the observed population genetic differentiation as well as connectivity patterns should be incorporated in the planning of future management activities such as translocations.
Population genetic structure of the common warthog (Phacochoerus africanus) in Uganda: evidence for a strong philopatry among warthogs and social structure breakdown in a disturbed population
(African Journal of Ecology, 2007) Muwanika, Vincent B.; Nyakaana, Silvester; Siegismund, Hans R.; Arctander, Peter
Fine-scale genetic structure of large mammals is rarely analysed. Yet it is potentially important in estimating gene flow between the now fragmented wildlife habitats and in predicting re-colonization following local extinction events. In this study, we examined the extent to which warthog populations from five localities in Uganda are genetically structured using both mitochondrial control region sequence and microsatellite allele length variation. Four of the localities (Queen Elizabeth, Murchison Falls, Lake Mburo and Kidepo Valley) are national parks with relatively good wildlife protection practices and the other (Luwero), not a protected area, is characterized by a great deal of hunting. In the total sample, significant genetic differentiation was observed at both the mtDNA locus (FST ¼ 0.68; P < 0.001) and the microsatellite loci (FST ¼ 0.14; P < 0.001). Despite the relatively short geographical distances between populations, significant genetic differentiation was observed in all pair-wise population comparisons at the two marker sets (mtDNA FST ¼ 0.21–0.79, P < 0.001; microsatellite FST ¼ 0.074–0.191, P < 0.001). Significant heterozygote deficiency was observed at most loci within protected areas while no significant deviation from Hardy–Weinberg expectation was observed in the unprotected Luwero population. We
A recent bottleneck in the warthog and elephant populations of Queen Elizabeth National Park, revealed by a comparative study of four mammalian species in Uganda national parks
(Animal Conservation, 2003) Muwanika, Vincent B.; Siegismund, Hans R.; Okello, John Bosco A.; Masembe, Charles; Arctander, Peter; Nyakaana, Silvester
Until 1972, Uganda’s national parks boasted of large numbers of large mammal species. Following the breakdown of law and order between 1972 and 1985, large-scale poaching led to an unprecedented decline in numbers of most large mammals in Uganda’s national parks. However, the extent of decline varied in the different parks across different animal species. We have investigated the genetic effects of these reductions in four mammalian species (the common warthog, African savannah elephant, savannah buffalo and common river hippopotamus) from the three major parks of Uganda using both microsatellite loci (for elephant and warthog populations) and mitochondrial control sequence variation in the warthogs, elephants, buffaloes and hippopotamuses. Queen Elizabeth National Park showed extreme reduction in nucleotide diversity for two species, the common warthog (π = 0.0%) and African elephant (π = 0.4%); no such decrease was found for the two other species, the buffalo (π = 3.7–5.4%) and hippopotamus (π = 1.7–1.9%), in the three parks. Nuclear microsatellite markers on the other hand showed high gene diversity in all populations in the common warthog (mean He 0.66–0.78) and the African savannah elephant (mean He 0.68–0.72). We interpret these results in terms of varying poaching pressure in the different parks, susceptibility of different species to poaching and differences in effective population sizes at the mitochondrial and nuclear loci
Six new polymorphic microsatellite loci isolated and characterized from the African savannah elephant genome
(Molecular Ecology Notes, 2005) Nyakaana, Silvester; Okello, John B. A.; Muwanika, Vincent B.; Siegismund, Hans R.
The African savannah elephant (Loxodonta africana) is a ‘keystone’ species that plays a vital role in regulating the dynamics of both plant and animal communities and yet it is endangered and its numbers have been reduced to approximately 500 000 across their entire continental range. Molecular genetic markers are important tools for providing genetic information useful in formulating effective management and conservation strategies for the surviving elephant populations. We describe the isolation and characterization of six new polymorphic microsatellite markers in the African savannah elephant and demonstrate that these loci can be PCR (polymerase chain reaction)-multiplexed, a desirable attribute that saves costs in large-scale microsatellite screening.
Three genetically divergent lineages of the Oryx in eastern Africa: Evidence for an ancient introgressive hybridization
(Conservation Genetics, 2006) Masembe, Charles; Muwanika, Vincent B.; Nyakaana, Silvester; Arctander, Peter; Siegismund, Hans R.
Phylogeographic and population genetic studies using sequence information are frequently used to infer species boundaries and history; and to assess hybridization and population level processes. In this study, partial mitochondrial DNA (mtDNA) control region (423 bp) and cytochrome b sequences (666 bp) of Oryx beisa sampled from five isolated localities in its entire current range in Africa were analyzed to investigate the extent of genetic variation and differentiation between populations. We observed high nucleotide diversity at the control region in the total sample (6.3%) but within populations, it varied considerably ranging from 1.6% to 8.1%. Population pairwise genetic differentiation was generally significantly high (ranging from FST = 0.15, P<0.01 to FST = 0.54, P<0.001). In the total sample, 29 and 12 haplotypes were observed in the control region and the cytochrome b data sets respectively. For both data sets, the haplotypes cluster into three distinct clades (sequence divergence ranged from 6.0%–12.9% to 0.8%–1.0% for the control region and cytochrome b sequences, respectively) that do not correspond to sampling locations. Two of these clades are found in the same localities (Samburu and Marsabit), which represent the O.beisa beisa subspecies, whereas the last clade represents the fringe-eared oryx (O. beisa callotis). We interpret these findings in terms of an ancient hybridization and introgression between two formerly isolated taxa of Oryx beisa.
Time Clustered Sampling Can Inflate the Inferred Substitution Rate in Foot-And- Mouth Disease Virus Analyses
(PLoS ONE, 2015) Pedersen, Casper-Emil T.; Frandsen, Peter; Wekesa, Sabenzia N.; Heller, Rasmus; Sangula, Abraham K.; Wadsworth, Jemma; Knowles, Nick J.; Muwanika, Vincent B.; Siegismund, Hans R.
With the emergence of analytical software for the inference of viral evolution, a number of studies have focused on estimating important parameters such as the substitution rate and the time to the most recent common ancestor (tMRCA) for rapidly evolving viruses. Coupled with an increasing abundance of sequence data sampled under widely different schemes, an effort to keep results consistent and comparable is needed. This study emphasizes commonly disregarded problems in the inference of evolutionary rates in viral sequence data when sampling is unevenly distributed on a temporal scale through a study of the foot-andmouth (FMD) disease virus serotypes SAT 1 and SAT 2. Our study shows that clustered temporal sampling in phylogenetic analyses of FMD viruses will strongly bias the inferences of substitution rates and tMRCA because the inferred rates in such data sets reflect a rate closer to the mutation rate rather than the substitution rate. Estimating evolutionary parameters from viral sequences should be performed with due consideration of the differences in short-term and longer-term evolutionary processes occurring within sets of temporally sampled viruses, and studies should carefully consider how samples are combined.

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