Ticks of domestic animals in Lesotho: Morphological and molecular characterization.

A total of 3311 tick specimens were randomly collected from domestic animals including cattle, sheep, goats, horses, donkeys, and dogs from Lesotho districts namely, Berea, Butha-Buthe, Leribe, Mafeteng, Maseru, Mohale's Hoek, Mokhotlong, Qacha's Nek, Quthing and Thaba Tseka. Tick species were identified morphologically and verified by amplification and sequencing of the CO1 and 18S rRNA genes. Nine species were identified under different genera namely, Haemaphysalis elliptica 0.1% (n = 2), Hyalomma rufipes 2.6% (n = 87), Hy. truncatum 1.2% (n = 41), Otobius megnini 13.6% (n = 451), Rhipicephalus appendiculatus 0.1% (n = 3), Rhipicephalus decoloratus 9.3% (n = 308), Rhipicephalus evertsi evertsi 65.1% (n = 2156), Rhipicephalus glabroscutatum 1.3% (n = 43) and Rhipicephalus microplus 6.6% (n = 220). There was a significant difference at p = 6.2E-06 (ꭓ2 = 1.923, df = 7) in the distribution of tick species and their abundance p = 0.04 (ꭓ2 = 1.923, df = 7) from each population. The CO1 and 18S rRNA sequences matched the morphological determinations on the NCBI database and clustered with relevant species on the phylogenetic tree. Genetic analysis of CO1 and 18S rRNA provided very strong support for monophyly of the Rhipicephalinae and Ornithodorinae complexes. Both CO1 and 18S rRNA are useful genetic markers for the specific and generic characterization of tick species in Lesotho and elsewhere. This is the first scientific publication of tick species occurring in Lesotho.

Highly pathogenic avian influenza (A/H5N1) virus outbreaks in Lesotho, May 2021.

In May 2021, Lesotho reported its first outbreak of highly pathogenic avian influenza (HPAI) to the OIE. Samples were collected from infected poultry and the virus was confirmed by molecular tests to be of the H5N1 subtype. Full genome sequencing and phylogenetic analysis revealed that the viruses belonged to clade 2.3.4.4b and showed high identity with A/H5N1 viruses identified in Nigeria and Senegal in early 2021. The identification of A/H5N1 HPAI in Lesotho has important implications for disease management and food security in the region.

Epidemiology of Rabies in Lesotho: The Importance of Routine Surveillance and Virus Characterization.

Rabies is widespread throughout Africa and Asia, despite the fact that the control and elimination of this disease has been proven to be feasible. Lesotho, a small landlocked country surrounded by South Africa, has been known to be endemic for rabies since the 1980s but the epidemiology of the disease remains poorly understood due to limited sample submission, constrained diagnostic capabilities, and a lack of molecular epidemiological data. Considering the existing challenges experienced in Lesotho, we aimed to evaluate the direct, rapid immunohistochemical test (DRIT) as an alternative to the direct fluorescent antibody (DFA) test for rabies diagnosis in Lesotho. Towards this aim, extensive training on the implementation and interpretation of the DRIT was hosted in Lesotho in April 2016 before both tests were applied to all samples subjected to routine rabies diagnosis at the Central Veterinary Laboratory (CVL). We found agreement between the DFA and DRIT assays in 90/96 samples (93.75%). The samples that produced inconsistent results (n = 6) were re-tested a further two times with both assays before being subjected to a real-time qPCR to confirm the diagnosis. Additionally, a statistically significant three-fold increase in the average number of samples submitted per month was observed after the DRIT implementation started, following continuous rabies awareness initiatives amongst the animal health professionals in the country over a 12-month period (p = 0.0279). Partial G-L intergenic regions of selected rabies-positive samples (n = 21) were amplified, sequenced, and subjected to phylogenetic analyses. Molecular epidemiological analyses, that included viruses from neighbouring provinces in South Africa, suggested that at least three independent rabies cycles within Lesotho were implicated in instances of cross-border transmission. This study has evaluated alternative methods for diagnosing and improving rabies surveillance in Lesotho, as well as providing new information that would be of importance in the planning of future disease intervention campaigns, not only in Lesotho, but also in neighbouring South Africa.