Multigene phylogeny of South African <i>Anopheles </i>mosquitoes

Until recently, no medicinal uses were recorded for the South African Erythroxylaceae species, although, this family is used world wide in traditional medicine. This study reveals for the first time that Erythroxylum delagoense and Erythroxylum pictum roots were used to treat dysentery and diarrhoea and that Erythroxylum emarginatum leaves decoction was used to treat asthma, kidney problems, arthritis, child bearing problems and influenza in South Africa. To validate some of the medicinal uses, antibacterial testing was done for the first time on all five South African species. Leaf and bark extracts of four of the five South African Erythroxylaceae species ( E. delagoense, E. emarginatum, E. pictum and Nectaropetalum capense ) showed some good antibacterial activities with MIC <1 mg/ml. E. delagoense showed good results against Bacillus subtilis , Klebsiella pneumoniae and S taphylococcus aureus ; E. emarginatum against Klebsiella pneumoniae ; E. pictum against Bacillus subtilis and Klebsiella pneumonia; N. capense against Klebsiella pneumonia . Key words: Antimicrobial activity, Erythroxylaceae, medicinal uses, South Africa.

South African Helichrysum species: A review of the traditional uses, biological activity and phytochemistry

Two South African Pyrgomorpha species have reduced chromosome numbers, due to centric fusions between the largest autosomes and the medium and small autosomes. P. rugosa has 2n♂=11(XO) (4 pairs of submetacentric and 1 pair of acrocentric autosomes) and P. granulata has 2n♂=13(XO) (3 pairs of submetacentric and 3 pairs of acrocentric autosomes). A third South African species has a typical Pyrgomorphidae number of 2n♂=19(XO) (acrocentrics). The mean chiasma frequency of the 2n♂=19 species is higher than that of the other two, although the frequencies of distal chiasmata in all three are similar. The recombination potential of the two species with lower chromosome numbers has been reduced, due to fewer crossovers in comparison to the 2n♂=19 species, as well as to independent assortment.

Marthasterias glacialis are found in the cool-temperate waters of the north-eastern Atlantic Ocean, in the subtropical waters of the Mediterranean Sea and along the south-western tip of Africa. The South African Marthasterias population includes two morphotypes, a smooth, spineless rarispina form and a spiny africana form, that have been described as separate species, subspecies, or forma by various authors over the past century. To test whether these two morphotypes represent separate species, and whether either, or both, are conspecific with the North-East Atlantic species, 78 Marthasterias were collected from the Cape Peninsula of South Africa. Morphological comparisons between individuals of the two forms showed no significant clustering of samples, indicating that there is no morphological separation of the two South African forms into distinct groupings. The africana and rarispina forms were also shown to be genetically indistinguishable, using a fragment of the mitochondrial cytochrome c oxidase subunit I (COI) gene and the nuclear internal transcribed spacer 1 (ITS1). In addition, the COI sequences were also compared to those from European specimens, and phylogenetic reconstruction and intra- and interspecific levels of divergence suggested that the South African specimens form a single group that is genetically distinct from the European M. glacialis. Although the allopatric distribution, high genetic divergence (more than 3% for the COI fragment) and morphological differences suggest that the South African form should be raised to species status under the name Marthasterias africana, further work must assess an independent genetic marker (nuclear) to support raising the COI clade to species level. True M. glacialis have a spine-armament pattern of a series of three or more regular spine rows down the length of each arm, whereas M. africana are either covered in many irregularly spaced spines, or have an extraordinarily bare surface with only two spine rows per arm. Marthasterias africana may also have an actinal spine simulating the presence of a third inferomarginal spine. This work tentatively resolves the taxonomic dispute, elucidates the separation and amalgamation of the two African forms and suggests a single, uniquely South African Marthasterias species that might be distinct from the north Atlantic M. glacialis, although further analyses to test reproductive isolation between the North-East Atlantic and South African forms are required.

Two Umbrina species, U. canariensis Valenciennes 1843 and U. robinsoni Gilchrist and Thompson 1908, are recognised from southern Africa. The latter species was hitherto believed to be a synonym of Umbrina ronchus Valenciennes 1843 (type locality Canary Islands). U. canariensis is distributed along the South Africa eastern seaboard from Cape Point to Sodwana Bay and U. robinsoni is known from False Bay to Madagascar and Oman. African Umbrina taxonomy has, however, been hindered by geographic samples that were either too few or consisted of specimens of disparate length; and as a result the identification and distribution of South African Umbrina species was confused. Morphological comparison of a large number of South African Umbrina with specimens from the type locality (Canary Islands) confirmed the identity of South African U. canariensis and allowed for an expanded description of the species. However, differences between specimens of U. ronchus and those of the second South African species (n = 251) led us to resurrect U. robinsoni (Gilchrist and Thompson 1908) as a valid name for this species. U. robinsoni differs from U. ronchus in having a smaller supraoccipital crest and thus a less steep pre-dorsal profile; a shallower preorbital bone (13–21% head length [HL] vs 21% HL); and a shorter nostril-orbit distance (2.4–6.9% HL vs 7.8–8.5% HL). Colour patterns also differ between the two species, with U. ronchus lacking the oblique, wavy, white stripes evident on the flanks of U. robinsoni. U. ronchus does not occur in South African waters, and is an eastern Atlantic species occurring from Gibraltar to Angola. Specimens from the east coast of Africa (Moçambique to Gulf of Oman) that were previously identified as U. ronchus are U. robinsoni. Differences between U. robinsoni and U. canariensis include: a lower modal number of soft dorsal fin rays, (22–27 vs 24–30); less deep body depth, (26–36% standard length [SL] vs 33–39% SL); shorter pectoral fin length (15–21% SL vs 20–25% SL); longer caudal peduncle length (26–34% SL vs 21–28% SL) and snout length (27–38% HL vs 23–32% HL); and smaller orbit diameter (14–33% HL vs 23–34% HL). Otoliths of U. robinsoni differ from those of U. canariensis in being smaller, less elongate, lacking a massive post-central umbo and having a post-dorsal spine remnant. The body colour and nature of the striping pattern on the flanks differs markedly between the species: in U. robinsoni the oblique stripes are thin, wavy, white lines; in U. canariensis the oblique stripes are thicker, nearly straight and brown; U. robinsoni also lacks the triangle-shaped mark on the outer operculum and the dark pigmentation of the inner operculum that is found on U. canariensis. Spatial analysis of South African specimens collected with a variety of gear revealed U. robinsoni to be a shallow-water species found from the surf-zone to 40m, whereas U. canariensis occurs predominantly from 40 to 100m depth. Although both species occur throughout the South African eastern seaboard, U. canariensis is most common west of the Kei River, where the shelf is wider. Examination of three specimens of U. steindachneri Cadenat 1950 confirmed the presence of a fourth sub-Saharan Umbrina species that is limited to tropical West African waters from Senegal to Angola. U. steindachneri differs from the other African Umbrina in having a high number of soft dorsal rays (28–29), a greater 3rd dorsal spine length (25–27% SL) and a very pronounced and convoluted striping pattern on the flanks.

Several species of Achnatherum (grass tribe Stipeae) and Melica (tribe Meliceae) typically are infected by nonpathogenic, seed-transmissible fungi with characteristics of Neotyphodium species (anamorphic Clavicipitaceae). Molecular phylogenetic studies clearly have distinguished the endophytes from Achnatherum inebrians (from Xinjiang Province, China), A. robustum and A. eminens (both from North America) and indicate that the A. inebrians endophyte comprises a unique nonhybrid lineage within the Epichloë and Neotyphodium phylogeny, whereas the endophytes of A. robustum, and A. eminens are hybrids with multiple Epichloë species (holomorphic Clavicipitaceae) as ancestors. Likewise distinct hybrid origins are indicated for Neotyphodium species from the European Melica species, M. ciliata and M. transsilvanica, the South African species M. decumbens and M. racemosa, and the South American species M. stuckertii. Neotyphodium species have been described from A. inebrians from Gansu Province, China, (N. gansuense), A. eminens (N. chisosum), M. stuckertii (N. tembladerae) and the South African Melica species (N. melicicola). However the endophytes from A. robustum and the European Melica species have not been described and the phylogenetic relationships of N. gansuense have not been investigated. Here we report a comprehensive study of morphological features and phylogenetic analyses of β-tubulin and actin gene sequences on an expanded collection of endophytes from the Stipeae and Meliceae. These data provide a firm foundation for the description of two new Neotyphodium species, N. guerinii from M. ciliata and M. transsilvanica, and N. funkii from A. robustum. We also propose the new variety, N. gansuense var. inebrians for endophytes of A. inebrians from Xinjiang Province, which are morphologically and phylogenetically distinct from, yet clearly related to, N. gansuense from Gansu Province.

Several species of Achnatherum (grass tribe Stipeae) and Melica (tribe Meliceae) typically are infected by nonpathogenic, seed-transmissible fungi with characteristics of Neotyphodium species (anamorphic Clavicipitaceae). Molecular phylogenetic studies clearly have distinguished the endophytes from Achnatherum inebrians (from Xinjiang Province, China), A. robustum and A. eminens (both from North America) and indicate that the A. inebrians endophyte comprises a unique nonhybrid lineage within the Epichloe and Neotyphodium phylogeny, whereas the endophytes of A. robustum, and A. eminens are hybrids with multiple EpichlooY species (holomorphic Clavicipitaceae) as ancestors. Likewise distinct hybrid origins are indicated for Neotyphodium species from the European Melica species, M. ciliata and M. transsilvanica, the South African species M. decumbens and M. racemosa, and the South American species M. stuckertii. Neotyphodium species have been described from A. inebrians from Gansu Province, China, (N. gansuense), A. eminens (N. chisosum), M. stuckertii (N. tembladerae) and the South African Melica species (N. melicicola). However the endophytes from A. robustum and the European Melica species have not been described and the phylogenetic relationships of N. gansuense have not been investigated. Here we report a comprehensive study of morphological features and phylogenetic analyses of beta -tubulin and actin gene sequences on an expanded collection of endophytes from the Stipeae and Meliceae. These data provide a firm foundation for the description of two new Neotyphodium species, N. guerinii from M. ciliata and M. transsilvanica, and N. funkii from A. robustum. We also propose the new variety, N. gansuense var. inebrians for endophytes of A. inebrians from Xinjiang Province, which are morphologically and phylogenetically distinct from, yet clearly related to, N. gansuense from Gansu Province.

In 2006, a strain of bluetongue virus serotype 8 (BTV-8) of sub-Saharan origin was responsible for the first outbreaks in recorded history of clinical bluetongue disease (BT) in northern Europe. In this study, we examine the oral susceptibility of Culicoides (Avaritia) imicola Kieffer (Diptera: Ceratopogonidae) and other livestock-associated Culicoides species from southern Africa to infection with several strains of BTV-8. Following feeding using an artificial membrane-based method and incubation, virus was found in <1% of C. imicola individuals tested. Higher rates of susceptibility were found, however, for a variety of other South African species, including Culicoides (Avaritia) bolitinos Meiswinkel. Although these results do not preclude the role of C. imicola as a vector of BTV-8, its low susceptibility to BTV indicates that other less abundant Culicoides species may have the potential to play decisive roles in the epidemiology of this virus and should not be excluded from risk assessment studies.

Genome size (C-value) and the percentage of adenine and thymine nucleotides in the genome (AT content) are fundamental characteristics of every species, and very important parameters in molecular cytogenetic and phylogenic studies, and for the progress of whole-genome sequencing. In this study, the C-value and AT content of nine abalone species inhabiting the Northwest Pacific (around Japan), Southeast Asia, South Africa, and Oceania were determined using flow cytometry. C-value ranged from 1.32 pg for Haliotis varia (Southeast Asia) to 2.01 pg for Haliotis laevigata (Oceania), and AT content ranged from 58.0% for Haliotis planata (Southeast Asia) to 66.3% for Haliotis diversicolor aquatilis (Southeast Asia). Reported chromosome numbers ranged from 2n = 32 to 36, and were higher in Oceanian, South African, and Northwest Pacific species (2n = 36) compared with the Southeast Asian group (2n = 32). This increase of chromosome number in the Oceanian species seems to be related to an increase in the DNA amount, unlike that of Haliotis midae (South African species). An increase of the C-value associated with an increase in DNA amount was also shown in the North Pacific group. These results suggest that increases in the DNA amount occurred independently in the ancestor of each of the different lineages.

By virtue of their low vagility, flightless insects are useful indicators of biogeographical history. Relationships of the flightless dung beetle genus, Gyronotus, are of particular interest due to its Gondwanaland ancestry, distinctive relict distribution along the south-eastern seaboard of Africa, and its restriction to forests which are seriously threatened by exploitation. Because of the limited number of diagnostic morphological characters, it was necessary to code morphometric data in order to conduct distance and cladistic parsimony analysis of interspecific relationships in Gyronotus. There was a good correlation between relationships indicated by the dendrograms/cladograms and those determined by an examination of aedeagus character states both of which indicate a disjunction between south and east African species and a broad separation between northern and southern South African species. Comparison of the bilaterally asymmetrical aedeagi of Gyronotus with the symmetrical aedeagi of the sister genus, Anachalcos Hope, suggests geographical polarization of character states from greater plesiomorphy in east African Gyronotus to greater apomorphy in South African species, particularly in the southernmost element in which the aedeagus shows extreme asymmetry. Furthermore, body shape follows a similar geographical gradient in that the three Gyronotus species of tropical east Africa are significantly more elongate than the three ovoid, lowland/afromontane species of South Africa. An examination of historical factors suggests that this spatially-restricted distribution is the relict of a very old tropical lowland pattern. In extant taxa, the phylogenetic polarization is towards one of five main centres of afrotropical forest biodiversity in the geologically old Eastern Arc and the adjoining lowland forest (Swahili centre of endemism). Survivors from old lineages may be one reason for such centres of high biodiversity.

The genus Salvia (Lamiaceae) encompasses about 900 species worldwide of which 26 are indigenous to southern Africa. The South African species are used in traditional medicines [1], as spice or tea due to reported good anti-oxidant properties [2]. The anti-oxidant capacity of these plants has been ascribed to the presence of phenolic compounds such as rosmarinic acid (RA), caffeic acid, carnosic acid and carnosol [2]. HPTLC-densitometric and HPLC-UV chromatographic techniques were used for qualitative and quantitative analyses of RA in 18 methanol:chloroform extracts from sixteen Salvia species. Polynomial and linear regression analyses were used to estimate the amount of RA in solvent extracts by HPTLC-densitometric and HPLC-UV techniques, respectively. RA was identified in all the samples investigated and ranged from 13.1µg/mg (S. stenophylla Burch. ex Benth.) to 113.0µg/mg (S. muirii L.Bolus). The paired sample t-test showed no statistical significant difference in the estimation of the amount of RA in the solvent extracts using the two chromatographic techniques. A strong correlation (r2=0.93) was found between the estimation using the HPTLC-densitometric and the HPLC-UV calibrations.

The Restionaceae is one of the three major families defining fynbos, the characteristic vegetation type of the Cape Floristic Region. Periodic fires with a frequency of 5 to 40 years are a natural phenomenon in fynbos vegetation. Fire-stimulated seed germination has been reported for a variety of fynbos species, and species in the Asteraceae, Ericaceae, Proteaceae and Restionaceae have shown a germination response to smoke and/or aqueous smoke extracts. In the present study seed of 32 species was screened to obtain an indication of how important the smoke cue is for germination in the South African Restionaceae. The results of the present study represents the first occasion that comparative germination data for South African species in this family have ever been obtained. Twenty-five of the 32 species tested showed a statistically significant improvement in germination following smoke treatment. Untreated seeds of 18 of the species responding, showed a high degree of dormancy with only 0.1% to 2.0% germination. These results suggest that under natural conditions smoke from fynbos fires may provide an important cue for triggering seed germination in this family. The degree of improvement in germination following smoke treatment ranged from 147% in the case of Restio festuciformis to 25300% in the case of Rhodocoma capensis. It is suggested that the 16 species which showed a 1000% or more increase in germination following smoke treatment form a group in which smoke is likely to be the major cue for germination. In those species in which there is a lesser response, smoke may be one of a number of germination cues which include heat, and possibly alternating high and low incubation temperatures. The four species that did not germinate were all myrmecochorus, nut-fruited species. More information is needed concerning the fire survival strategy of South African Restionaceae species and many more will have to be investigated in the nursery and in the field, before the full pattern of response within the family and its significance can be elucidated.

The perceived cosmopolitanism of polychaete worms could be an artefact of historical factors such as poor original species descriptions, lack of type material and the European taxonomic bias, to name a few. Thus, it is possible that several cosmopolitan species hide complexes of cryptic and pseudocryptic species. Two putative cosmopolitan species, Platynereis dumerilii and Platynereis australis, collected in South Africa were investigated here (1) to determine whether the South African taxa are conspecific with the morphologically identical taxa from France and New Zealand (the respective type localities of P. dumerilii and P. australis), (2) to compare the South African species morphometrically to determine whether their morphological characters are reliable enough to separate them, and (3) to investigate whether these species have geographically structured populations along the coast of South Africa. Molecular data (COI and ITS1) confirm that P. dumerilii and P. australis do not occur in South Africa. Instead, the South African taxon formerly thought to be Platynereis dumerilii is new and is described here as Platynereis entshonae, sp. nov.; the identity of the other South African species is currently unresolved and is treated here as Platynereis sp. Surprisingly, Platynereis massiliensis (type locality: Marseilles) nested within the South African Platynereissp. clade but, since it is part of a cryptic species complex in the Mediterranean, the name is considered doubtful. Morphological characters traditionally used to define these South African Platynereis species are not reliable as predefined morphological groupings do not match phylogenetic clades and principal component scores revealed no separation in morphological characters that could distinguish between them. Haplotype networks and phylogenetic trees revealed that P. entshonae, sp. nov. and Platynereis sp. have geographically structured populations along the South African coast. http://zoobank.org/urn:lsid:zoobank.org:pub:6E36A210-9E48-430F-8A93-EDC27F0C5631

Chromosome numbers are presented for five genera in the Hemimerideae. Mitotic counts were made for 69 species of Diascia, seven species of Nemesia, six species of Alonsoa, four species of Hemimeris, and two species of Diclis. At the species level, 80 of these counts are new reports. Counts for two genera (Diclis and Hemimeris) are also new. Diascia has a base number of x = 9 and most species are diploid. Four tetraploid and one hexaploid species were discovered among the perennials of section Racemosae, bringing the total number of polyploids for the section to seven of 26 species (ca. 27%). Two species in section Racemosae have more than one cytotype; D. patens has 2n and 4n populations, whereas D. capsularis has 2n, 4n and 6n populations. In section Diascia, consisting entirely of annuals, only five of 46 species (ca. 11%) are tetraploid, while the rest are diploid. Polyploidy has apparently had multiple origins in both sections of Diascia. The seven species of Nemesia counted were uniformly 2n = 18, but hypotheses of a close relationship between Nemesia and Diascia conflict with differences in corolla aestivation and preliminary molecular evidence. Hemimeris and Diclis are diploid with 2n = 14 and appear to be closely related. Alonsoa is also related to Hemimeris with South African species having 2n = 28 and South American species mostly 2n = 56. Chromosomal, pollination and morphological differences between South African and South American species suggest that Alonsoa originated in South Africa and dispersed to South America before or shortly after the breakup of Gondwanaland. If true, the Scrophulariaceae has had a more ancient presence in the Southern Hemisphere than previously suspected.

The phylogeny of the early African hominins has long been confounded by contrasting interpretations of midfacial structure. In particular, the anterior pillar, an externally prominent bony column running vertically alongside the nasal aperture, has been identified as a homology of South African species Australopithecus africanus and Australopithecus robustus. If the anterior pillar is a true synapomorphy of these two species, the evidence for a southern African clade of Australopithecus would be strengthened, and support would be given to the phylogenetic hypothesis of an independent origin for eastern and southern African "robust" australopith clades. Analyses of CT data, however, show that the internal structure of the circumnasal region is strikingly different in the two South African australopith species. In A. africanus the anterior pillar is a hollow column of cortical bone, whereas in A. robustus it is a column of dense trabecular bone. Although Australopithecus boisei usually lacks an external pillar, it has internal morphology identical to that seen in A. robustus. This result supports the monophyly of the "robust" australopiths and suggests that the external similarities seen in the South African species are the result of parallel evolution.