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Lumpy skin disease is one of the fast-spreading viral diseases of cattle and buffalo that can potentially cause severe economic impact. Lesotho experienced LSD for the first time in 1947 and episodes of outbreaks occurred throughout the decades. In this study, eighteen specimens were collected from LSD-clinically diseased cattle between 2020 and 2022 from Mafeteng, Leribe, Maseru, Berea, and Mohales' Hoek districts of Lesotho. A total of 11 DNA samples were analyzed by PCR and sequencing of the extracellular enveloped virus (EEV) glycoprotein, G-protein-coupled chemokine receptor (GPCR), 30 kDa RNA polymerase subunit (RPO30), and B22R genes. All nucleotide sequences of the above-mentioned genes confirmed that the PCR amplicons of clinical samples are truly LSDV, as they were identical to respective LSDV isolates on the NCBI GenBank. Two of the elevem samples were further characterized by whole-genome sequencing. The analysis, based on both CaPV marker genes and complete genome sequences, revealed that the LSDV isolates from Lesotho cluster with the NW-like LSDVs, which includes the commonly circulating LSDV field isolates from Africa, the Middle East, the Balkans, Turkey, and Eastern Europe.

Haemonchus contortus is the most pathogenic and economically restrictive gastrointestinal nematode in the small ruminant industry globally. Morbidity, poor cross-bodily state, and mortality of sheep in Lesotho suggest the presence of H. contortus. The present study investigated the morphological, molecular, and population genetics of H. contortus third-stage larvae infecting sheep in four ecological zones (EZ) of Lesotho. Coprocultures were prepared for larval morphological identification and PCR determination. Larvae were identified morphologically as 100% H. contortus. The Second Internal Transcribed Spacer (ITS-2) gene of the ribosomal DNA of H. contortus isolates in the present study revealed nucleotide homology ranging from 97 to 100% when compared with selected GenBank reference sequences. Pairwise evolutionary divergence among H. contortus isolates was low, with 0.01318 recorded as the highest in the present study. Five haplotypes resulted from 14 Lesotho sequences. Haplotype diversity and nucleotide diversity were 0.76923 and 0.00590, respectively. Genetic differentiation among isolates was low but not statistically significant. An analysis of molecular variance revealed that most molecular variation was distributed within topographic populations at 94.79% (F ST  = 0.05206, p > 0.05) and 5.21% among populations. There was high gene flow and no definite population genetic structure among Lesotho isolates., Competing Interests: Declaration of Competing Interest The authors do not declare any competing interests., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Free-range chickens are an integral part of poultry production in many developing countries. In the Mountain Kingdom of Lesotho, the majority of the population own free-range chickens, which serve a variety of purposes including being a source of meat, eggs and use for cultural rituals amongst others. However, there is lack of scientific studies on occurrence of parasitic infections on free-range chickens in Lesotho. The aim of this study was to document common gastrointestinal parasites infecting free-range chickens in four villages of Mafeteng District in Lesotho. A total number of 462 pooled faecal samples were collected from various households in HaKubutu (n = 114), HaMatjeka (n = 120), HaMpalipali (n = 120) and Thabang Villages (n = 108) which were subjected to microscopic examination using McMaster technique. The prevalence of gastrointestinal parasite infection was as follows: Eimeria tenella (12.8%), Ascaridia galli (10.4%) and Heterakis gallinarum (5%). The prevalence for H. gallinarum and Ascaridia galli were comparatively higher during the hot-wet season (7.1% and 2.8% respectively) than the cold-dry season (3.2% and 1.9% respectively) and varied significantly (P < 0.05). For E. tenella, the oocysts per gram were slightly higher in the cold-dry season than the hot-wet season. Polymerase chain reaction only amplified DNA from six (29%) adult A. galli worms of which two amplicons were successfully sequenced. The obtained cytochrome C oxidase subunit 1 partial gene sequences displayed 98-100% identity with South African A. galli isolates. This is the first scientific study on prevalence and molecular characterization of nematodes and coccidia species infecting free-range village chickens in Lesotho. The findings can be used to review management of gastrointestinal nematodes and protozoal parasites of free-range chickens in Lesotho., Competing Interests: Declaration of Competing Interest The authors have no conflicts of interest that might unduly influence their work., (Copyright © 2022 Elsevier B.V. All rights reserved.)

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 (ꭓ 2  = 1.923, df = 7) in the distribution of tick species and their abundance p = 0.04 (ꭓ 2  = 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., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Tick-borne diseases (TBDs) hamper the growth of the livestock sector and impose major constraints for the health and management of domestic animals in the tropic and subtropical regions globally. Currently, there is no scientific report on the presence of zoonotic pathogens transmitted by tick species in Lesotho. This study aimed to identify zoonotic tick-borne pathogens of economic importance from ticks infesting domestic animals in Lesotho using molecular techniques. A total of 322 tick DNA pools were subjected to PCR screening for the presence of zoonotic pathogens and sequenced. The overall prevalence of Anaplasma spp. was 35% (113/322), with a 100% infection rate in Rhipicephalus microplus , followed by R . evertsi evertsi (92%), Hyalomma rufipes and Otobius megnini sharing 50% and the lowest infection rate was observed in R . decoloratus with 40%. The prevalence of Coxiella burnetii , a gram-negative pleomorphic etiological agent of Query fever (Q fever), was 1% (2/322) for all screened samples, with 20% of R . decoloratus and 1% of R . e . evertsi infected. Rickettsia africae was detected from Hyalomma rufipes with a 70% prevalence. This study provides a baseline knowledge of tick-borne pathogens of medical and veterinary importance in Lesotho and raises awareness of the prevalence of such diseases within the tourism sector as they are mostly affected.