Numerous outbreaks of high-pathogenicity avian influenza (HPAI) were reported during 2020–2021. In Africa, H5Nx has been detected in Benin, Burkina Faso, Nigeria, Senegal, Lesotho, Namibia and South Africa in both wild birds and poultry. Botswana reported its first outbreak of HPAI to the World Organisation for Animal Health (WOAH) in 2021. An H5N1 virus was detected in a fish eagle, doves, and chickens. Full genome sequence analysis revealed that the virus belonged to clade 2.3.4.4b and showed high identity within haemagglutinin (HA) and neuraminidase proteins (NA) for viruses identified across a geographically broad range of locations. The detection of H5N1 in Botswana has important implications for disease management, wild bird conservation, tourism, public health, economic empowerment of vulnerable communities and food security in the region.

We sought to investigate whether SARS‐CoV‐2 was present, and to perform full‐length genomic sequencing, in a 5‐year‐old male crossbreed dog from Gaborone, Botswana that presented overt clinical signs (flu‐like symptoms, dry hacking cough and mild dyspnoea). It was only sampled a posteriori, because three adult owners were diagnosed with SARS‐CoV‐2 infection. Next‐generation sequencing based on Oxford Nanopore Technology (ONT) was performed on amplicons that were generated using a reverse transcriptase real‐time polymerase chain reaction (RT‐qPCR) of confirmed positive SARS‐CoV‐2 nasopharyngeal and buccal swabs, as well as a bronchoalveolar lavage with mean real cycle threshold (qCt) value of 36 based on the Nucleocapsid (N) gene. Descriptive comparisons to known sequences in Botswana and internationally were made using mutation profiling analysis and phylogenetic inferences. Human samples were not available. A near‐full length SARS‐CoV‐2 genome (∼90% coverage) was successfully genotyped and classified under clade 20 O and Pango‐Lineage AY.43 (Pango v.4.0.6 PLEARN‐v1.3; 2022‐04‐21), which is a sublineage of the Delta variant of concern (VOC) (formerly called B.1.617.2, first detected in India). We did not identify novel mutations that may be used to distinguish SARS‐CoV‐2 isolates from the dog and humans. In addition to Spike (S) region mutation profiling, we performed phylogenetic analysis including 30 Delta sequences publicly available reference also isolated from dogs. In addition, we performed another exploratory analysis to investigate the phylogenetic relatedness of sequence isolated from dog with those from humans in Botswana (n = 1303) as of 31 March 2022 and of same sublineage. Expectedly, the sequence formed a cluster with Delta sublineages – AY.43, AY.116 and B.1.617.2 – circulating in same time frame. This is the first documented report of human‐associated SARS‐CoV‐2 infection in a dog in Botswana. Although the direction of transmission remains unknown, this study further affirms the need for monitoring pets during different COVID‐19 waves for possible clinically relevant SARS‐CoV‐2 transmissions between species.

We sought to investigate whether SARS-CoV-2 was present, and to perform full-length genomic sequencing, in a 5-year-old male crossbreed dog that presented with flu-like symptoms (including a dry hacking cough and mild dyspnea) and resided in a household with 3 adults that were diagnosed with SARS CoV-2 infection. Next generation sequencing based on MinION technology was performed on amplicons that were generated using a reverse transcriptase real-time polymerase chain reaction (RT-qPCR) of confirmed positive SARS-CoV-2 nasopharyngeal and buccal swabs, as well as a bronchoalveolar lavage with mean qCt value of 36 based on the Nucleocapsid gene. Descriptive comparisons to known sequences in Botswana and internationally were made using mutation profiling analysis and phylogenetic inferences based on maximum likelihood. Samples from the dog’s owners were not available. A near-full length SARS-CoV-2 genome (~90% coverage) was successfully genotyped and classified under clade 20 O and Pango-Lineage AY.43 (Pango v.4.0.6 PLEARN-v1.3; 2022-04-21), which is a sub-lineage of the Delta variant of concern (VOC) (formerly called B.1.617.2, first detected in India). We did not identify novel mutations that may be used to distinguish SARS-CoV-2 isolates from the dog and humans. In addition to S region mutation profiling, we performed phylogenetic analysis using Delta sequences from Botswana (n=1303); expectedly, the sequence isolated from the dog was closely related to the Delta sequences, particularly the AY.43, AY.116, and B.1.617.2 sub-lineages that were reported in Botswana within the same time frame. This is the first documented report of human-associated SARS-CoV-2 infection in a dog in Botswana. Although the direction of transmission remains unknown, this study further affirms the need for monitoring pets during different COVID-19 waves for possible clinically relevant SARS-CoV-2 transmissions between species.

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