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Summary Stegelletina salinaria is redescribed from Spain and South Africa. Spanish material was collected in sandy soil from the type locality and a nearby area, while one population collected in compost was examined from South Africa. The species is characterised by its small size (0.27-0.41 mm), lateral field with three incisures, lip region with three pairs of dentate lips bearing four tines each, primary axils bearing a large guarding process, secondary axils lacking guarding processes, nerve ring and secretory-excretory pore at isthmus level, spermatheca and post-vulval uterine sac poorly developed, female tail conoid with rounded hyaline terminus and males unknown. The description, light micrographs and scanning electron microscope images, illustrations and molecular analyses are provided. 18S and 28S rDNA sequence analyses show that the isolates of S. salinaria from Spain and South Africa are closely related to each other and to other species of Stegelletina, while also indicating a close relationship with some species of Nothacrobeles, Scottnema and Stegelleta (28S tree) and Zeldia (18S tree).

Kiwifruit (Actinidia species), a temperate plant, is becoming one of the most important crops in tropical regions of high altitudes at Limpopo Province, South Africa. During a survey on free-living nematodes in five kiwifruit orchards, 19 nematode species were identified from the soil samples across five kiwi orchards at Limpopo Province. Overall, the most prevalent nematodes encountered were Rhabditis species, followed by Cephalobus persegnis and Aporcelaimellus with 46.37, 45.12 and 18.29% frequency of occurrence, respectively. Prominence Values (PVs) of Rhabditis sp. (17834.40) and C. persegnis (14329.70) were higher when as compared to those of other free-living nematode species. The results indicated no significant correlation between the free-living nematode species and pH, except for Plectus aquatilis. However, Ca% had a significant positive correlation with C. persegnis, Acrobeles complexus, Clarkus papillatus, Trypilina, Diptherophora, Prismatolaimus and Aporcelaimellus. In contrast, Na and Ca exhibited a significant positive correlation with P. rigidus. In conclusion, the high altitude-kiwifruit-producing regions suggested the existence of a considerable range of free-living nematodes, with the highest nematode diversity in Nooyenskopje orchards.

Soil inhabiting organisms are important determinants of agroecosystem productivity. Understanding the composition, the abundance, and the type of interactions established by soil microorganisms is therefore crucial to design strategies to improve agricultural practices and agroecosystem management. In this study, we collected Zeldia punctata nematodes in maize fields in South Africa and profiled their associated bacterial communities using next-generation sequencing. We observed that Z. punctata nematodes establish associations with ecologically diverse bacterial species. The most abundant species observed are Pseudomonas syringae, a phytopathogenic bacterial complex; Lactobacillus paraplantarum, a broadly distributed bacterial species that is present in soils, water bodies, and animal intestinal tracts and has certain probiotic and antimicrobial properties; and Melissococcus plutonius, a serious pathogenic bacterial species that causes brood disease in honeybees. Our study contributes to a better understanding of the soil bacterial communities associated with nematodes in maize agricultural soils in South Africa and unravels the presence of diverse detrimental and beneficial nematode-associated bacteria.

Ten Steinernema abbasi isolates were obtained during a soil survey in Western Uttar Pradesh, India. The nematode isolates were characterised using morpho-taxometrical and molecular approaches, and ...

Summary Butlerius butleri was isolated during a survey on free-living soil nematodes from South Africa. The South African population of B. butleri is characterised by having a large stoma divided into two chambers and bearing a thorn-like dorsal tooth, spicules 44 (41-49) μm long, gubernaculum 29 (28-30) μm long, three pairs of precloacal papillae and five pairs of postcloacal papillae, and a filiform tail 280 (204-332) μm long in females, 310 (256-356) μm long in males. The SEM observation showed the cheilostom is divided into six conical plates 2.1-2.3 μm long. Nblast of 18S rDNA revealed 99% similarity and 6-7 bp differences with the B. butleri (KP453998) from Iran. Molecular analysis of B. butleri based on the sequence of the 18S and 28S rDNA placed it together with Butlerius and close to Pseudodiplogasteroides and Diplogasteroides. This species showed predatory and cannibalistic behaviours, feeding on other B. butleri in the substrate. SEM photographs of the species are presented for the first time.

Summary A nematode survey conducted in the Telperion Nature Reserve (Mpumalanga, South Africa) resulted in a new Ironus species being described as well as two known species reported for the first time from South Africa, viz., Chronogaster aspinata and Paraphanolaimus behningi. Ironus telperionensis n. sp. is characterised by a long (1724 (1456-1864) μm) and slender body (50 times longer than mid-body diam.), long stoma (116 (87-129) μm), and a 235 (212-257) μm long tail with a ventral curl at the tip. The new species is morphologically most similar to I. ignavus and I. paludicola. However, the new species can be separated from I. ignavus based on the lower ratio c (7.3 (6.5-8.0) vs 8.0-11.0) and longer stoma (116 (87-129) vs 76-90 μm). Ironus telperionensis n. sp. can be distinguished from I. paludicola by the lower ratio a (49.2 (37.4-53.0) vs 60.0-80.0) and lower ratio c (7.3 (6.5-8.0) vs 8.0-13.0). The 18S rDNA phylogenetic analysis of I. telperionensis n. sp. placed it close to I. elegans (KC133064), whereas the 28S rDNA phylogenetic analysis placed it close to I. tenuicaudatus. The phylogenetic analysis revealed that Ironus is a monophyletic group. Paraphanolaimus behningi was identified based on anteriorly bent vagina in the female, spicule length (106 or 110 μm) and number of supplements (13 or 14) in the male. Chronogaster aspinata was identified based on body length (1127 or 1203 μm), cephalic setae length (7 μm), finely rounded tail terminus without a mucron or additional spines, and no longitudinal ridges. It fits the known description of the species. Although various differences were observed between previously reported populations of these species, the morphometrics of the specimens found in Telperion widen the existing variation in these two species.

Summary The population of Xiphinema americanum species group was detected from the soil surronding the roots of peach (Prunus persica) during a survey of plant-parasitic nematodes in Gilgit-Baltistan, Pakistan. From the results of the morphological study, the species was identified as X. diffusum. Morphometrics of X. diffusum generally agree with those of the type species and the topotype specimens. Molecular analysis of 18S rDNA of X. diffusum from Pakistan indicated two nucleotide differences and 99 % similarity with the Chinese (AM086669) and the Australian (AM086685) population of X. diffusum. Phylogenetic analysis of 18S rDNA placed the Pakistanis population close to X. diffusum from China and Australia. This species is reported for the first time from Pakistan

In a comprehensive survey of plant-parasitic nematodes of Gilgit Province, Pakistan, Quinisulcius capitatus (Allen, 1955) Siddiqi, 1971, was detected from 10 localities at high population density (≥1000 nematodes/100 g of soil) infecting different crop plantations. Morphplogical and molecular variations using principle component analysis of Q. capitatus have been given in this research. This species is characterized by body length (605–856 μm in females and 536–662 μm in males), stylet length (13–19 μm), pharynx (114–174 μm), tail length (40–56 μm in females and 38–50 μm in males), spicules (16–22 μm) and gubernaculum (6–8 μm). Principal component analysis (PCA) revealed the morphometric variation among the Pakistani populations of Q. capitatus. Phylogenetic analysis using the D2-D3 segment of 28S rDNA and ITS rDNA placed the Pakistani population of Q. capitatus in a clade with 100 posterior probability supports. Table with measurements, line illustrations, LM photographs, PCA plot and phylogenetic position for Q. capitatus within Dolchodoridae is given.

Entomopathogenic nematodes (EPNs) are important biocontrol agents of insect pests. To increase the availability of locally adapted entomopathogenic nematode isolates for biocontrol programs, a survey of several agricultural soils in Western Uttar Pradesh, India was conducted. Eight hundred and sixty soil samples from the districts Meerut, Bulandshahr, Baghpat, and Bijnor were collected and examined for the presence of entomopathogenic nematodos using the “Galleria baiting method”. Steinernema and Heterorhabditis nematodes were recovered. The isolated Heterorhabditis nematodes were molecularly, and morphologically characterized, and their biocontrol potential was evaluated against Spodoptera litura. Finally, the geographical distribution of entomopathogenic nematodes was studied based on the analysis of ITS GenBank records. A small proportion of the collected soil samples were positive for Heterorhabditis and Steinernema nematodes. Twelve soil samples were positive for the presence of Heterorhabditis nematodes, and 29 samples were positive for Steinernema. The Heterorhabditis nematodes were identified as Heterorhabditis indica based on morphological, morphometrical and molecular analyses. No other species of Heterorhabditis were isolated from the soil samples analyzed, suggesting that this species is dominant in the western part of Uttar Pradesh, India. The morphology of the nematode isolates was somewhat similar to the morphology of the H. indica isolate used for the original description of this species, with a notable exception mucrons were present in the hermaphrodite and female specimens we collected, but this structure was not observed in the specimens used for the original description of the species. Principal component analyses (PCA) show small inter- and intraspecific morphological variability between the nematodes species of the “Indica” clade. The insecticide properties of one isolate, CH7, were evaluated against Spodoptera litura, and the results show that this isolate effectively killed this pest under laboratory conditions, demonstrating its potential as a biocontrol agent. This study sets the basis for establishing new biocontrol agents to be used in future pest management programs in India.

Summary Paratrophurus anomalus is redescribed from the North-West Province, South Africa, associated with the rhizosphere of a willow tree, a dominant ornamental tree in the province. This population is characterised by its adult body length (696 (625-834) μm for female and 706 μm for male), lateral field with four longitudinal incisures, cephalic framework well cuticularised, lip region smooth and lacking annuli, female stylet 19.6 (18-21) μm long, female tail cylindrical, 35 (31-38) μm long, c′ = 2.3 (2.1-2.8), with thick hyaline region forming 31-43% of the tail length, spicules 22 μm long and gubernaculum 12 μm long. Morphologically, P. anomalus is very similar to P. kenanae and P. dissitus, although they differ on the basis of lip region morphology, stylet length, hyaline tail region, and spicule length. In addition, SEM observations are provided for the first time for this species along with a new host and new geographical record of the species from South Africa, providing new information for the revision of Paratrophurus. Molecular analysis of P. anomalus using ITS rDNA showed a close relationship with P. bursifer, P. bhutanensis and Bitylenchus species. In addition, principal component analysis was done for 14 character states of species in the genus.

Alternatives to hazardous insecticides are urgently needed for an environmentally friendly and effective management of insect pests. One such option is the use of entomopathogenic nematodes (EPN). To increase the availability of EPN with potential for biocontrol, we surveyed agricultural soils in the Republic of Rwanda and collected two Steinernema isolates. Initial molecular characterization showed that they represent a new species, for which we propose the name S. africanum n. sp. To describe this new species, we reconstructed phylogenetic relationships, calculated sequence similarity scores, characterized the nematodes at the morphological level, conducted crossing experiments, and isolated and characterized their symbiotic bacteria. At the molecular level, S. africanum n. sp. is closely related to S. litorale and S. weiseri. At the morphological level, S. africanum n. sp. differs from closely related species by the position of the nerve ring and also because the stoma and pharynx region is longer. The first-generation males have ventrally curved spicules with lanceolate manubrium and fusiform gubernaculum and the second-generation males have rounded manubrium and anteriorly hook-like gubernaculum. Steinernema africanum n. sp. does not mate or produce fertile progeny with any of the closely related species.

During a survey in Iran and South Africa on the plant parasitic nematode, a criconematid species namely, Criconema mutabile was recovered from undisturbed land. Criconema mutabile is characterised by having body length 240–378 μm long, with total body annuli 88–118, conspicuous lip region offset from the body with one annulus; long and robust stylet 56–76 μm, long and short rounded tail, 12–19 μm long. Illustration and measurements for both Iranian and South African populations and LM and SEM photographs of the South African population are provided. Phylogenetic analysis of the Iranian and South African C. mutabile using 28S rDNA, placed them together with populations of the same species from Venezuela (AY780954; 11 nucleotide difference; 98% identity) and Iran (KU722385; 7 nucleotide difference; 99% identity). In addition, the population studied showed 100% identity with two populations of unidentified Criconema (FN433874 and FN433865) from California, USA.

Soil inhabiting organisms are important determinants of agroecosystem productivity. Understanding the composition, the abundance, and the type of interactions established by soil microorganisms is therefore crucial to design strategies to improve agricultural practices and agroecosystem management. In this study, we collected Zeldia punctata nematodes in maize fields in South Africa and profiled their associated bacterial communities using next-generation sequencing. We observed that Z. punctata nematodes establish associations with ecologically diverse bacterial species. The most abundant species observed are Pseudomonas syringae, a phytopathogenic bacterial complex; Lactobacillus paraplantarum, a broadly distributed bacterial species that is present in soils, water bodies, and animal intestinal tracts and has certain probiotic and antimicrobial properties; and Melissococcus plutonius, a serious pathogenic bacterial species that causes brood disease in honeybees. Our study contributes to a better understanding of the soil bacterial communities associated with nematodes in maize agricultural soils in South Africa and unravels the presence of diverse detrimental and beneficial nematode-associated bacteria.

After hatch, second-stage juveniles (J2) of root-knot (Meloidogyne species) nematodes could spend at least 12 weeks in soil solutions searching for penetration sites of suitable host plants. The external covering of nematodes, the cuticle, consists of various layers that contain glycoproteins, lipids, soluble proteins (collagens) and insoluble proteins (cuticulins). Generally, cucurbitacins are lipophilic, but there is scant information on how cuticular proteins relate to these complex terpenoids. A study was conducted to investigate the nature and extent of damage post-exposure of J2 to a wide range of Nemafric-BL phytonematicide concentrations. Post-72 h exposure to Nemafric-BL phytonematicide, nematode morphometrics versus phytonematicides exhibited either negative quadratic, positive quadratic, or negative linear relations, with the models explained by significant (P

During a survey on the biodiversity of plant-parasitic nematodes of natural areas in Botswana, Mesocriconema sphaerocephalum was discovered around the rhizosphere of the wild grass. The nematodes were extracted using the tray method and then fixed according to the available protocols. The morphological characters fit well with the M. sphaerocephalum. Besides, molecular aspects using 18S and 28S rDNA were studied. The phylogenetic analysis of 18S and 28S rDNA placed the examined population with other populations of M. sphaerocephalum in a group. According to the knowledge, this is the first report of M. spaherocephalum from Botswana.

Agricultural soils and open fields from Western Uttar Pradesh (India) were surveyed to determine the presence of entomopathogenic nematodes. From the entomopathogenic nematodes isolated, Heterorhabditis isolates were selected and further characterized using morphological, morphometrical and molecular approaches. The results showed that three isolated nematodes were Heterorhabditis bacteriophora and were associated with Photorhabdus laumondii subsp. clarkei bacteria, while the rests were identified as Heterorhabditis indica. The biocontrol potential of H. bacteriophora against three agricultural pests was evaluated. Nematode infectivity experiments showed that the nematode isolates DH7 and DH8 were highly pathogenic against cotton bollworm (Helicoverpa armigera) and tobacco cutworm (Spodoptera litura), and less pathogenic against white grub (Holotrichia serrata) larvae. This study sets the basis for establishing new biocontrol agents to be used in pest management programs in India.

Entomopathogenic nematodes (EPNs) are important biocontrol agents of insect pests. To increase the availability of locally adapted entomopathogenic nematode isolates for biocontrol programs, a survey of several agricultural soils in Western Uttar Pradesh, India was conducted.Eight hundred and sixty soil samples from the districts Meerut, Bulandshahr, Baghpat, and Bijnor were collected and examined for the presence of entomopathogenic nematodos using the "Galleria baiting method". Steinernema and Heterorhabditis nematodes were recovered. The isolated Heterorhabditis nematodes were molecularly, and morphologically characterized, and their biocontrol potential was evaluated against Spodoptera litura. Finally, the geographical distribution of entomopathogenic nematodes was studied based on the analysis of ITS GenBank records.A small proportion of the collected soil samples were positive for Heterorhabditis and Steinernema nematodes. Twelve soil samples were positive for the presence of Heterorhabditis nematodes, and 29 samples were positive for Steinernema. The Heterorhabditis nematodes were identified as Heterorhabditis indica based on morphological, morphometrical and molecular analyses. No other species of Heterorhabditis were isolated from the soil samples analyzed, suggesting that this species is dominant in the western part of Uttar Pradesh, India. The morphology of the nematode isolates was somewhat similar to the morphology of the H. indica isolate used for the original description of this species, with a notable exception mucrons were present in the hermaphrodite and female specimens we collected, but this structure was not observed in the specimens used for the original description of the species. Principal component analyses (PCA) show small inter- and intraspecific morphological variability between the nematodes species of the "Indica" clade. The insecticide properties of one isolate, CH7, were evaluated against Spodoptera litura, and the results show that this isolate effectively killed this pest under laboratory conditions, demonstrating its potential as a biocontrol agent.This study sets the basis for establishing new biocontrol agents to be used in future pest management programs in India.

Helicotylenchus species were recovered from natural areas of three provinces (Mazandaran, Kerman and Guilan) in Iran, with morphological and molecular characteristics suggesting that they were Helicotylenchus minzi and Helicotylenchus pseudorobustus. A phylogenetic analysis of the two Helicotylenchus species using 28S rDNA placed H. pseudrobustus and H. minzi in a clade with 1.00 posterior probability. Results suggest that the Helicotylenchus population identified using molecular characteristics as H. pseudorobustus is a paraphyletic group, with hierarchical clustering analysis indicating a close relationship between the Iranian and the Chinese populations of H. pseudorobustus. In conclusion, further molecular studies within the genus Helicotylenchus are needed, especially for morphologically similar populations that have been recognized as separate species.

Basiria gracilis is redescribed from North West Province, South Africa, associated with willow (Salix sp.), which is a dominant ornamental tree species in the province. B. gracilis is characterised by its female body length (692–994 µm), lateral field with two longitudinal incisures, weakly cuticularised labial framework, high lip region lacking annuli, stylet 8–13 µm long, axial spermatheca filled with small rounded sperm, conical-elongated female tail with round to pointed terminus (103–146 µm, c=6.1–7.4, c’=8.8–12.1). Morphologically, B. gracilis is closely related to B. paragracilis but the taxa are differentiated on the basis of lip region and stylet morphology. In addition, B. gracilis is reported for the first time from South Africa. Line drawings, LM and SEM illustrations are given for the species.

Kiwi is becoming one of the most important fruit in subtropical regions of South Africa with altitudes that confer sufficient chilling requirements. During a survey on biodiversity of plant-parasitic nematodes of kiwi in Magoebaskloof in Limpopo Province, several plant-parasitic nematodes were discovered, with Meloidogyne species occurring at the highest frequency. Nematodes were sampled from roots and the rhizosphere of one stunted Kiwi tree, extracted using the tray method and then fixed. The morphological characters fit well with those of M. hapla. The molecular approach using ITS and 28S rDNA, along with the related phylogenetic analysis, placed the examined population in a group with other populations of M. hapla. Kiwi is being reported as a new host for M. hapla in South Africa.

Shelf-life information provides end-users with the assurance that the product is still in compliance with label claims. Behavioral reaction orders of the Arrhenius model had been consistently used under fixed conditions to provide shelf-life in food products. Due to non-conformity of the cucurbitacin-containing phytonematicides to the Arrhenius behavioral reaction orders, an alternative quadratic model consistent with the behavioral reaction orders of cucurbitacins was developed under chilled (5°C at 95-98% RH) and fixed tropical (38°C at 90% RH) conditions, while room temperature constituted unfixed conditions. Sampling for cucurbitacins was done at time-frames compliant with geometric series, with cucurbitacin analysis regularly performed using high-performance liquid chromatography techniques. Under chilled conditions, neither the Arrhenius nor the quadratic model could predict the shelf-life for Nemarioc-AL phytonematicide, whereas Nemafric-BL phytonematicide had shelf-life of 35 weeks. In contrast, under tropical conditions, the positive quadratic models showed that Nemarioc-AL and Nemafric-BL phytonematicides had shelf-life of 35 and 825 weeks, respectively. In conclusion, the two phytonematicides could be stored under fixed tropical conditions to enhance the shelf-life of their active ingredients.

Third-stage larvae (L3) of Steinernema feltiae exist as free-living infective juveniles (IJ), with suspended development activities. In contrast, parasitic stages (L1, L2, L4, adult) have mutualistic relations with Xenorhabdus species bacteria, along with unique morphological changes and development inside the cadaver of host insects and/or plant-parasitic nematodes. Commercial IJ strains are tolerant to cucurbitacin-containing phytonematicides, but we have scant information on how morphological adjustments in IJ are achieved. In this study, we investigated the nature of morphological adjustments in commercial S. feltiae IJ strains to Nemafric-BL phytonematicide, which contains cucurbitacin B as active ingredient. Post-72 h exposure to phytonematicide concentration, IJ specimens were fixed on mounting slides. Length (body, excretory pore to anterior end, pharynx, rectum, stoma, tail), diameter (head width, neck base, mid-body, anal body), cuticle thickness and De Man ratios were measured with a computer software programme attached to Omax light microscope. Morphometric data against increasing phytonematicide concentration exhibited either density-dependent quadratic, linear or neutral relations. Increase in body length at low phytonematicide concentration was accompanied by decrease in tail length and pharynx length during muscle contraction when IJ were still alive. After death at high phytonematicide concentration, the opposite morphometric effects ensued due to muscle relaxation. The observed changes in morphometric structures were explained on the basis of morphological adjustments that modulated volumes of pseudocoelom cavity in IJ. The modulation is intended to maintain hydrostatic pressure within permissible upper limits in order to avoid structural damage to internal organs embedded in the pseudocoelom fluids.

During a survey of plant-parasitic nematodes in Iran, five populations of Pratylenchus neglectus were studied by means of morphological and molecular characterization. The results showed that body length has high significant correlation with morphometric data indices such as a (r = 0.487; p ≤ 0.01), b (r = 0.355; p ≤ 0.05), c (r = 0.527; p ≤ 0.01), stylet (r = 0.625; p ≤ 0.01), excretory pore (r = 0.750; p ≤ 0.01), overlapping (r = 0.434; p ≤ 0.01) and tail length (r = 0.447; p ≤ 0.05). However, the highest correlation was detected between pharynx overlapping and postvulval uterine sac (r = −0.935; p ≤ 0.01). A molecular study of the P. neglectus populations using 28S rDNA showed close relationships of the Iranian, UK (KX683377), South Korea (KY468842) and China (JX046968). The COI of mtDNA analysis showed close relationship of the Iranian P. neglectus with the Chineese populations (KX349423; KY424105). The results indicated monophyly of P. neglectus. Illustration and measurement tables are provided for the species.

During a survey of soil nematodes in Iran, some known species belonging to the genus Ditylenchus were recovered from the rhizosphere of alfalfa: Ditylenchus acutus, Ditylenchus myceliophagus and Di...

Summary A population of Macrolaimus arboreus was collected from the rhizosphere of pine in Iran. This population of M. arboreus is characterised by the body length, 800-1010 μm in females, lip region with six seta-like papillae directed anteriad, stoma nearly as long as wide (9-11 × 7 μm), with cheilostom slightly longer than gymnostom and cheilorhabdia and gymnorhabdia well-developed, pharyngeal corpus 1.8-2.0 times the isthmus length, with slightly swollen metacorpus, excretory pore located at isthmus level, female reproductive system monodelphic-prodelphic reflexed with short post-vulval uterine sac, female tail conical (57-67 μm, c = 13.7-15.0, c′ = 3.5-3.8), ending in an acute tip which is weakly curved dorsad, and phasmids at 54-60% of tail length. Measurements and illustrations, including SEM, are provided.

SummaryIn a survey of nematode damage in groundnut (Arachis hypogaea) (Fabaceae) production areas in the Vaalharts Irrigation Scheme in the Northern Cape Province of South Africa, a nematode was recovered and initially believed to be a member of the genusRobustodorus,i.e., the species has a characteristic robust stylet with very well-developed teardrop-shaped (garlic bulb-like) basal swellings. The specimens were recovered in large numbers from damaged hulls and kernels of field-collected groundnuts and, after thorough morphological and molecular studies, were subsequently identified asAphelenchoides arachidis. Molecular analysis based on near-full-length small subunit (SSU), internal transcribed spacer (ITS) and D2-D3 expansion segments of the large subunit (LSU) rRNA genes supported the monophyly ofA. arachidis,A. subtenuisandR. megadoruswithin the subfamily Aphelenchoidinae. Based on the typological characters observed in the present study compared with descriptions in the literature, these three species are considered as congeneric,i.e., they share very well-developed basal swellings of the stylet. Accordingly, these twoAphelenchoidesspecies are transferred toRobustodorusasR. arachidisn. comb. andR. subtenuisn. comb. In addition, the male tail characters ofRobustodorusandAphelenchoidesare discussed.

Hirschmanniella anchoryzae from Iran and Pratylenchus hippeastri from South Africa were recovered during a survey of plant-parasitic nematodes belonging to the family Pratylenchidae. Both species were studied using morphological and molecular techniques. Hirschmanniella anchoryzae is identified based on the flattened head, short stylet (19–22 µm), excretory pore position (anterior to pharyngo-intestinal junction), spicule length (27–30 µm), and existence of an axial mucro at the tail end. Phylogenetic analysis using 28S rDNA showed monophyly of Hirschmanniella which Iranian H. anchoryzae placed close to H. halophila (EU620464; EU620465). This result was supported by the principal component analysis of Hirschmanniella species. SEM observation of the South African population of P. hippeastri showed the presence of two annuli in the lip region. Morphometric characters resembled those of specimens earlier reported from South Africa. Hierarchal cluster using morphometrical criteria showed that the Floridian (USA) and South African populations form a group. However, the principal component analysis showed variation within this species. The molecular study of P. hippeastri populations using 18S, ITS, 28S rDNA, and COI of mtDNA showed that all P. hippeastri cluster in one group and confirmed the identification of the species using both morphological and molecular techniques. In addition, the results indicated that South African populations group close to the USA populations. Illustrations of both species including light and scanning electron microscopy observations for P. hippeastri are provided.

The entomopathogenic nematodes have been reported from all continents (except Antarctica) and almost all regions of the world. Surveys of EPNs in India has resulted in the recovery of several isolates of Steinernema. Among one of them, isolate CS34 was identified as S. hermaphroditum Stock, Griffin & Chaerani, 2004. We investigated the identification and the pathogenicity of S. hermaphroditum in District Meerut of Western Uttar Pradesh, India. The Steinernema was examined for its pathogenicity and accurate identification by the mean of morphological and molecular technique and its geographical distribution was mapped based on meta-analysis of the ITS GenBank records. The surveys of agricultural soils of district Meerut, India, resulted in the isolation of one strain from entomopathogenic nematode labelled CS34 through Galleria baiting technique. Morphological characters and morphometrical analysis indicated that the strain CS34 was closely related to the “glaseri” group of Steinernema spp. The Nblast results indicated that ITS rDNA sequence had no nucleotide differences in comparison with the S. hermaphroditum (JQ687355). However, one variation in the D2–D3 segment of 28S rDNA was observed in comparison with the AY598358. The phylogenetic analysis using ITS and 28S rDNA indicated that the Indian S. hermaphroditum could be placed together with other S. hermaphroditum, with strong posterior probability. Besides, the PCA analysis demonstrated some variability within the test populations. The distribution of S. hermaphroditum based on meta-analysis of the GenBank records showed its presence in the three Asian countries—India, Thailand and Indonesia. The Indian strain of S. hermaphroditum also tested positively for its virulence against three major pests, namely, Galleria mellonella, Helicoverpa armigera, and Spodoptera litura, with resultant which showed good efficacy on the mortalities. In conclusion, the economy of India is agriculture-based, but there are huge losses due to different insect pests infesting different crops. Steinernema hermaphroditum CS34 is an indigenous species to Indian subcontinent and efforts should be made to evaluate its virulence and pathogenicity against the other agricultural pests hampering productivity throughout the country. This may lead to incorporate S. hermaphroditum strain CS34 as a regular biological control agent against important lepidopteran pest in integrated pest management programs in the future.