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Rinus Knoetze, Antoinette Swart and Louwrens R. Tiedt

A new cyst nematode, herein described as Globodera capensis n. sp., was found on several farms in the Swartland and Sandveld areas in South Africa. As the host plant of G. capensis n. sp. is currently unknown, the species is represented by cysts and second-stage juveniles (J2) only. The cysts have a spherical form, are yellow to dark brown in colour and have bullae (vulval bodies) and subcuticular punctations. Six to 20 cuticular ridges/lines are present between the anus and vulval basin, Granek’s ratio is 0.7-4.0, the vulval basin diam. is 16-28.5 μm and vulval basin to anus distance is 19-60 μm. The J2 is 430-528 μm long and the stylet is 23-28 μm long with anteriorly flattened to rounded stylet knobs. These stages are morphologically and morphometrically very near to, or indistinguishable from PCN, G. artemisiae, G. millefolii and G. tabacum tabacum. It can be distinguished by the molecular characteristics of the ITS-region of the ribosomal DNA. Pairwise distances between closely related Globodera species show that G. capensis n. sp. differs the least in number of base differences per sequence from G. millefolii (63-64 bp) and most from an undescribed Globodera from Chile (72-73 bp). Phylogenetic relationships of G. capensis n. sp. with selected species of Punctoderinae, inferred from ITS-rRNA sequences by using the Maximum Likelihood method, indicate that G. capensis n. sp. occupies a basal position within a lineage of Globodera species from Europe, Asia and New Zealand that parasitises non-solanaceous plants. For diagnostic purposes, digestion of the ITS1 with AluI will distinguish G. capensis n. sp. from G. mexicana, G. pallida, G. rostochiensis and G. tabacum, but not from G. artemisiae, G. millefolii and G. zelandica, whilst FauI will distinguish G. capensis n. sp. from all the other Globodera species tested.

Antoinette P. Malan, Rinus Knoetze and Louwrens R. Tiedt

Steinernema nguyeni n. sp. was recovered by baiting from beneath an Olea africana tree in South Africa. The combination of morphological and molecular features suggests that S. nguyeni n. sp. is a member of the feltiae-kraussei-oregonense group, clustering with members of this group in Clade III. The new species is morphologically characterised by the infective juvenile body length of 737 (673-796) μm and the number of ridges in the infective juvenile lateral field is 2, 8, 2. The male of the first generation can be recognised by the spicule length of 66 (58-75) μm and a gubernaculum length of 43 (30-55) μm. The first generation female can be recognised by the vulval lips only slightly protruding and the presence of low, double-flapped epiptygmata. Analysis of the ITS and D2-D3 regions of the ribosomal DNA confirms that S. nguyeni n. sp. differs from all other known Steinernema species.

Esther van den Berg, Louwrens R. Tiedt and Sergei A. Subbotin

During recent collections in South Africa, two species of Criconemoides were found. Criconemoides brevistylus is described and illustrated from a golf course in KwaZulu-Natal Province. It is compared with several closely related species, and three previously described species (C. helicus, C. onostris and C. paronostris) are regarded as junior synonyms of C. brevistylus. Criconemoides obtusicaudatus is described and illustrated from a maize field in Limpopo Province. The male of the species is described for the first time. Molecular characterisation of C. brevistylus and C. obtusicaudatus using the D2-D3 expansions segments of 28S rRNA and ITS rRNA gene sequences are provided. Phylogenetic relationships of these species with other Criconemoides are discussed.

Esther Van den Berg, Esther Van den Berg, Louwrens R. Tiedt, Esther Van den Berg, Louwrens R. Tiedt and Sergei A. Subbotin

Pin nematodes of the genus Paratylenchus are widely distributed across the world and associated with many plant species. Morphological identification of Paratylenchus species is a difficult task because it relies on many characters with a wide range of intraspecific variation. In this study we provide morphological and molecular characterisation of several pin nematodes: Paratylenchus aquaticus, P. dianthus, P. hamatus, P. nanus and P. straeleni, collected in different states of the USA and South Africa. Paratylenchus aquaticus is reported from South Africa and Hawaii and P. nanus is found from South Africa for the first time. Morphological descriptions, morphometrics, light and scanning electron microscopic photos and drawings are given for these species. Molecular characterisation of nematodes using the D2-D3 of 28S rRNA and ITS rRNA gene sequence revealed that samples morphologically identified as P. aquaticus, P. hamatus and P. nanus indeed represent species complexes containing several species. Sequences of the rRNA genes are also provided for several unidentified Paratylenchus. Phylogenetic relationships within the genus Paratylenchus are given as inferred from the analyses of the D2-D3 of 28S rRNA and ITS rRNA gene sequences. We present here the most complete phylogenetic analysis of the genus.

Isaiah Nthenga, Rinus Knoetze, Shaun Berry, Louwrens R. Tiedt and Antoinette P. Malan

A new species of entomopathogenic nematode, Steinernema sacchari n. sp., was isolated by trapping with the sugar cane borer, Eldana saccharina, from soil of a sugar cane field in the KwaZulu-Natal province of South Africa. The new species is morphologically characterised by the length of the infective juvenile (IJ) of 680 (630-722) μm, tail length of 64 (51-74) μm, ratio a = 19 (14-23), H% = 49 (43-57) and E% = 82 (70-109). The pattern of the lateral field of the IJ of the new species is 2, 5, 2 ridges (3, 6, 3 lines or incisures). The male of the first generation can be recognised by the long spicule of 83 (73-89) μm, gubernaculum of 61 (50-68) μm, D% = 67 (54-88) and GS% = 73 (66-81). The first generation male lacks a mucron, while the second generation male always has one. The first generation female can be recognised by the vulval lips not being raised, the possession of long double-flapped epiptygmata and the lack of a postanal swelling. Analysis of the ITS and D2D3 regions showed S. sacchari n. sp. to differ from all other Steinernema species and to belong to a new monophyletic group, the ‘Cameroonian’ clade, consisting of S. cameroonense, S. nyetense and S. sacchari n. sp. This group is closely related to the feltiae-kraussei-oregonense Clade III.

Willem P. Steyn, Rinus Knoetze, Louwrens R. Tiedt and Antoinette P. Malan

During a survey in the Mpumalanga province of South Africa, a Steinernema species was isolated from a soil sample taken from a litchi orchard. Steinernema litchii n. sp. can be separated from other, closely related, species in the glaseri-group by morphological, morphometric and molecular analyses. The infective third-stage juvenile of the new species has a body length of 1054 (953-1146) μm, distance from head to excretory pore of 78 (64-86) μm, as well as eight ridges (i.e., nine lines) in the mid-body region. The c-ratio of 10 (9-13) is low and the tail is long at 95 (73-105) μm. First generation males have a spicule length of 86 (76-96) μm and a gubernaculum length of 65 (59-72) μm. The tail of the first generation male lacks a mucron; that of the second generation always bears one. The genital papillae total 23 and consist of 11 pairs and an unpaired precloacal papilla. The vulva of S. litchii n. sp. has a slightly asymmetrical protuberance and short, double-flapped epiptygmata. The female has a slightly protuberant postanal swelling. Phylogenetic analysis of the internal transcribed spacer (ITS) and of the 28S (D2-D3) regions of the ribosomal DNA (rDNA) confirmed the close relationship of S. litchii n. sp. to the Karii-clade. Both morphological and molecular evidence support the species status of S. litchii n. sp.

Esther Van den Berg, Louwrens R. Tiedt, Jason D. Stanley, Renato N. Inserra and Sergei A. Subbotin

The genus Scutellonema contains more than 40 species of spiral nematodes with enlarged phasmids called scutella. In this study, we provide morphological and molecular characterisation of S. clavicaudatum sp. n., S. brachyurus, S. bradys, S. cavenessi, S. transvaalense, S. truncatum and Scutellonema sp. A. from North and Central America, and Africa. The new species, S. clavicaudatum sp. n., was found on sugarcane in South Africa and is characterised by a lack of lip annuli as in S. africanum, S. siamense and S. truncatum. The lip region, in both males and females, is conical and marked by six large rectangular blocks separated or fused with the submedian and lateral lip sectors, which surround a round and distinct labial disc. Females of this new species also have large vaginal glands, a functional spermatheca, the lateral field posterior to the scutellum ending in a bluntly pointed shape and a clavate tail. Morphological descriptions, measurements, light and scanning electron microscopic photos and drawings are also given for S. bradys, S. cavenessi, S. transvaalense and S. truncatum. The study of spiral nematode samples from Florida, USA, confirmed the presence of a morphologically and genetically atypical populations of S. bradys. The morphology of the S. bradys population from Bermuda grass in pasture land from central Florida fits that of type specimens of this species, but differs in having a truncate tail terminus rather than round and also a prominent spermatheca filled with flagellate spermatozoa. Other Florida Scutellonema samples analysed in this study belonged to S. cavenessi, a species native to West Africa. This is the first report of S. cavenessi in Florida, where it parasitises the ornamental plant Sansevieria trifasciata. Our study showed a high level of intraspecific variation for Scutellonema rRNA and mtDNA genes, which can reach 5.6% for the D2-D3 of 28S rRNA, 12.9% for the ITS rRNA genes and 14.4% for the COI gene. Phylogenetic relationships within Scutellonema are given as inferred from the analyses of the D2-D3 of 28S rRNA, ITS rRNA and the COI mtDNA gene sequences.

Annika Pieterse, Louwrens R. Tiedt, Antoinette P. Malan and Jenna L. Ross

Worldwide interest in Phasmarhabditis originates from the successful commercialisation of P. hermaphrodita as a biological control agent against molluscs in Europe. To date, P. hermaphrodita has not been isolated from South Africa and, therefore, the formulated product may not be sold locally. During a survey for mollusc-associated nematodes, P. papillosa was dissected from the slug, Deroceras reticulatum, collected from George, South Africa. The nematode was identified using a combination of morphological, morphometric, molecular and phylogenetic techniques. Virulence tests were conducted which demonstrated that P. papillosa caused significant mortality to the European invasive slug Deroceras panormitanum. Additional data are provided in the morphometrics of the infective juvenile and in the molecular identification, using the mitochondrial cytochrome c oxidase subunit I (cox1) gene. This is the first report of P. papillosa from the African continent and of its virulence against D. panormitanum.

Moses M. Lesufi, Antoinette Swart, Alex H. Mc Donald, Rinus Knoetze, Louwrens R. Tiedt and Mariette Truter

Aphelenchoides arachidis is reported for the first time from South Africa and for the fourth time outside Nigeria. The A. arachidis-infested pods from South Africa showed the following symptoms: small seeds with the testa wrinkled and darker in colour than that of non-infested seeds; the pods showed dark lesions and some seeds within the pods showed early germination. Differences between the two South African and the Nigerian populations of A. arachidis include more lateral lines in some specimens (2-4 vs 2) and, on average, longer post-uterine sac length (extending for 74 (41-96) and 62 (33-82) vs about 50% of vulva to anus distance). Scanning electron micrographs of this species are presented for the first time. The ITS regions of ribosomal DNA were amplified, sequenced, aligned and compared with other sequences of Aphelenchoides species. Two pathogenic fungi, Thielaviopsis basicola and Neocosmospora vasinfecta, were also isolated from this material.

Agil Katumanyane, Antoinette P. Malan, Louwrens R. Tiedt and Brett P. Hurley


Two isolates of Steinernema bertusi n. sp. were separately recovered from Tito, Mpumalanga, and Port Edward, Kwa Zulu Natal, South Africa. In this paper, we describe the isolates as a new entomopathogenic nematode (EPN) species using molecular and morphological methodologies. The new species belongs to the cameroonense-clade, which consists of nematodes only isolated from the African continent. Steinernema bertusi n. sp. is characterised by having the longest infective juvenile (IJ) for this clade at 716 (628-814) μm. The IJ is further characterised by a body diam. of 32 (28-36) μm and the pattern for the arrangement of the lateral ridges from head to tail is 2, 4, 5, 4, 2. The first-generation male spicule and gubernaculum length is 82 (72-88) μm and 63 (54-72) μm, respectively. Only 25% of the second-generation males possess a mucron. The first-generation females of S. bertusi n. sp. have a slightly protruding vulva, with double-flapped epiptygmata and a mucron at the posterior end. The new EPN species is most closely related to S. sacchari and is the sixth species to be included in the cameroonense-clade.