The prevalence and genetic characterisation of Cryptosporidium isolates from cattle in Kiruhura district, South Western Uganda.

Cryptosporidium is an emerging opportunistic zoonotic pathogen that causes diarrheal illness in a wide range of hosts including livestock and humans. This study set out to establish the prevalence of Cryptosporidium as well as the circulating genotypes in order to elucidate the potential role of cattle in the spread of human cryptosporidiosis. Rectal coprological samples from 363 cattle in 11 households in Kiruhura district, Southwestern Uganda were collected and screened for the presence of Cryptosporidium oocysts using the phenol auramine staining method followed by fluorescent microscopy. DNA was extracted from the microscopy positive samples and the COWP gene amplified using PCR. PCR products were sequenced and subjected to phylogenetic analysis. Additionally a multiplex realtime PCR was used to identify the Cryptosporidium spp. Multivariable mixed effect logistic regression models were used to identify potential risk factors for Cryptosporidium infection. The overall prevalence of Cryptosporidium was 7.7% (95% CI 5.1-10.9), and herd level prevalence was 33.3% (95% CI 18.5-52.2). We found a statistically significant difference (OR = 30.78, 95% CI 4.31-219.95, p = 0.001) between infection in bulls as compared to cows. There was no significant difference in the prevalence among the different cattle breeds sampled. All the sequenced COWP gene DNA amplicons were confirmed to be C. hominis, with 93%-100% identity to sequences in the GenBank. The amplification of the small subunit rRNA by multiplex realtime PCR further established that the isolates in this study are C. hominis. This study represents the first time naturally occurring C. hominis has been detected from cattle in Uganda.

Prevalence of Trypanosoma congolense and Trypanosoma vivax in Lira District, Uganda.

Trypanosomes are the causative agents of animal African trypanosomiasis (AAT) and human African trypanosomiasis (HAT), the former affecting domestic animals prevalent in Sub-Saharan Africa. The main species causing AAT in cattle are T. congolense, T. vivax, and T. b. brucei. Northern Uganda has been politically unstable with no form of vector control in place. The return of displaced inhabitants led to the restocking of cattle from AAT endemic areas. It was thus important to estimate the burden of trypanosomiasis in the region. This study was designed to compare the prevalence of animal African trypanosomes in cattle in Lira District using microscopy and polymerase chain reaction amplification (PCR) methods. In this cross-sectional study, a total of 254 cattle from the three villages of Acanakwo A, Barropok, and Acungkena in Lira District, Uganda, were selected by simple random sampling technique and screened for trypanosomiasis using microscopy and PCR methods. The prevalence of trypanosomiasis according to microscopic results was 5/254 (2.0%) as compared to 11/254 (4.3%) trypanosomiasis prevalence according to PCR analysis. The prevalence of trypanosomiasis infection in the animal studied is 11/254 (4.3%). Trypanosoma congolense was the most dominant trypanosome species with a proportion of 9/11 (81.8%), followed by T. vivax 1/11 (9.1%) and mixed infection of T. congolense/T. vivax1/11 (9.1%). Barropok village had the highest prevalence of trypanosomiasis with 6/11 (54.5%). There is a statistically significant relationship (OR = 6.041; 95% CI: 1.634-22.331; p < 0.05) between abnormal PCV and trypanosome infection. Polymerase reaction amplification was the most reliable diagnostic method due to its high sensitivity and specificity as compared to the conventional microscopic method. Polymerase reaction amplification appears to have adequate accuracy to substitute the use of a microscope where facilities allow. This study, therefore, underscores the urgent need for local surveillance schemes more especially at the grassroots in Uganda to provide data for reference guideline development needed for the control of trypanosomiasis in Uganda.

Molecular detection and phylogenetic analysis of lumpy skin disease virus from outbreaks in Uganda 2017-2018.

BACKGROUND: Lumpy skin disease (LSD) is an infectious viral disease of cattle caused by a Capripoxvirus. LSD has substantial economic implications, with infection resulting in permanent damage to the skin of affected animals which lowers their commercial value. In Uganda, LSD is endemic and cases of the disease are frequently reported to government authorities. This study was undertaken to molecularly characterize lumpy skin disease virus (LSDV) strains that have been circulating in Uganda between 2017 and 2018. Secondly, the study aimed to determine the phylogenetic relatedness of Ugandan LSDV sequences with published sequences, available in GenBank. RESULTS: A total of 7 blood samples and 16 skin nodule biopsies were screened for LSDV using PCR to confirm presence of LSDV nucleic acids. PCR positive samples were then characterised by amplifying the GPCR gene. These amplified genes were sequenced and phylogenetic trees were constructed. Out of the 23 samples analysed, 15 were positive for LSDV by PCR (65.2%). The LSDV GPCR sequences analysed contained the unique signatures of LSDV (A11, T12, T34, S99, and P199) which further confirmed their identity. Sequence comparison with vaccine strains revealed a 12 bp deletion unique to Ugandan outbreak strains. Phylogenetic analysis indicated that the LSDV sequences from this study clustered closely with sequences from neighboring East African countries and with LSDV strains from recent outbreaks in Europe. It was noted that the sequence diversity amongst LSDV strains from Africa was higher than diversity from Eurasia. CONCLUSION: The LSDV strains circulating in Uganda were closely related with sequences from neighboring African countries and from Eurasia. Comparison of the GPCR gene showed that outbreak strains differed from vaccine strains. This information is necessary to understand LSDV molecular epidemiology and to contribute knowledge towards the development of control strategies by the Government of Uganda.