This paper presents the first comprehensive annotated checklist of the spider fauna in the Witsand Kalahari Nature Reserve in the Northern Cape, South Africa. Historically, 45 species were sampled in the reserve. Recent collecting efforts since 2017 included pitfall trapping surveys, implementation of a rapid sampling protocol, and ad hoc collecting in various microhabitats. This enabled the collection of a further 162 species, taking the currently known diversity to 207 species in 40 families. The rapid sampling protocol showed that profes-sional arachnologists collected higher spider abundance and species richness than students for active methods (beating and litter sifting) but not pitfalls. Total abundance declined from the woodland (n=349) to the dunefield (n=251) and rocky outcrop (n=196), whereas species richness was similar between the dunefield (S=45) and woodland (S=43) and slightly lower on the rocky mountainside (S=38). Litter sifting was the most effective method and pitfall trapping the least. Among the recently collected specimens, 77 specimens were submitted for DNA barcoding, with the sequence data publicly available.
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