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Abstract

Coffee is an important commodity for Kenya, where production is steadily declining, despite a global rise in demand. Of the various constraints affecting production, plant-parasitic nematodes are a significant, but often overlooked, threat. As a perennial crop, treating plantations once infected with nematodes becomes difficult. The current study evaluated the drenching application of two biocontrol agents, Trichoderma asperellum and Purpureocillium lilacinum, for their nematode control efficacy, as well as their impact on the soil nematode community structure on mature, established coffee trees in Kenya. Seven Arabica coffee field trials were conducted over two years on trees of various ages. All the fields were heavily infested with Meloidogyne hapla, the first report of the species on coffee in Kenya. Both fungal biocontrol agents were detected endophytically infecting roots and recovered from soil but not until six months after initial applications. The population densities of M. hapla had significantly declined in roots of treated trees 12 months after the initial application, although soil nematode density data were similar across treatments. Based upon the maturity index and the Shannon index, treatment with T. asperellum led to improved soil health conditions and enrichment of diversity in the microbial community. Application of P. lilacinum, in particular, led to an increased abundance of fungivorous nematodes, especially Aphelenchus spp., for which P. lilacinum would appear to be a preferred food source. The soils in the trials were all stressed and denuded, however, which likely delayed the impact of such treatments or detection of any differences between treatments using indices, such as the functional metabolic footprint, over the period of study. A longer period of study would therefore likely provide a better indication of treatment benefits. The current study positively demonstrates, however, the potential for using biologically based options for the environmentally and climate-smart management of nematode threats in a sustainable manner on established, mature coffee plantations.

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Abstract

Potato Cyst Nematodes (PCN) (Globodera rostochiensis and G. pallida Woll.) are quarantine pests of potato (Solanum tuberosum L.) worldwide capable of causing significant yield loss and difficult to manage with conventional methods. The study explored the diversity of antagonistic fungi associated with PCN obtained from soil samples collected in Nyandarua and Nakuru Counties in Kenya and their effect on PCN egg viability and hatching was also evaluated. Twelve fungal isolates from five genera were isolated and characterized using morphological and molecular techniques. The twelve isolates were Trichoderma asperellum (4), T. hamatum, T. breve, T. atrobruneum, Amanita basiorubra, Setophoma terrestris (2), Penicillium chrysogenum and Clonostachys rosea. The most abundant isolate was Trichoderma spp. with 58 % occurrence. The effect of seven of the isolates on PCN eggs showed that T. breve and P. chrysogenum reduced egg viability by 41 % and 34 %, respectively while T. asperellum and T. breve reduced their hatching by 50 % on average. Trichoderma atrobrunneum, T. hamatum, and A. basiorubra also reduced the PCN egg viability by 27 % on average. These fungal isolates could provide a potential tool for PCN management in potato production systems for improved yields. However, further studies are warranted to validate these findings under greenhouse and field conditions. A more comprehensive bioprospecting survey for PCN associated antagonistic fungi needs to be extended to other potato growing regions to explore further cyst pathogens.

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Abstract

The root-burrowing nematode, Radopholus similis, is reputedly the most damaging nematode pest of banana and responsible for major production losses. In this study, the endophytic potential of 13 fungal isolates was assessed for the management of R. similis in East African Highland bananas (‘Ng’ombe’). All isolates successfully colonised tissue-cultured banana roots, with isolates from Trichoderma, Fusarium and Hypocrea producing the highest (⩾49.1%) and Beauveria isolates the lowest (⩽14.4%) colonisation. The fungal endophytes T. asperellum (ICIPE 700) and H. lixii (ICIPE 697) were the most effective in reducing R. similis densities (>81%) relative to the non-inoculated control. However, the combined inoculation of ICIPE 700 and ICIPE 697 led to greater suppression of R. similis (>21%) relative to individual inoculation. Suppression of R. similis following inoculation of banana roots with ICIPE 700 and/or ICIPE 697 was associated with the significant upregulation of the defence-related gene PR-1, the cell signalling gene calmodulin Ca2+ and the cell-wall-strengthening gene β-1,3-glucan synthase. This study demonstrates the potential for nematode management in bananas with fungal endophytes, especially using the isolates ICIPE 700 and ICIPE 697 when combined.

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Abstract

The legume cavalcade, Centrosema pascuorum, is used extensively as a cover crop and as a component of conservation agriculture systems. It is also an attractive rotation or cover crop for the management of root-knot nematodes (RKN; Meloidogyne spp.) as it is a non-host. RKN are persistent pests that are well known to be difficult to control. However, the mechanisms governing the non-host status of cavalcade is unknown. The current study established that cavalcade leaves are toxic to RKN as either aqueous extracts or soil amendments. Bioassays conducted using Meloidogyne javanica showed that a 90% concentration of aqueous extract derived from 1-month-old cavalcade leaves (89 mg crude extract ml−1) suppressed nematode hatch (82.9%) and killed infective second-stage juveniles of M. javanica (85.3%). Soil amendments with 1% (w/w) of 1-month-old cavalcade leaves (0.99 mg crude extract g−1 soil) also provided effective control of M. javanica in the glasshouse on okra. One-month-old leaves appeared more effective than 2- or 3-month-old leaves. The soil amendments had no adverse phytotoxic effect on okra seed germination. Our study demonstrates the potential for using cavalcade leaves or extracts to manage RKN. This may be due to the nematicidal activity of the various compounds in the leaves, such as flavonoids, phenols and terpenoids, which should be further assessed.

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Abstract

The banana weevil (Cosmopolites sordidus) and the burrowing nematode Radopholus similis represent two of the most important pests of bananas. Previously, colonization of banana plants by the non-pathogenic Fusarium oxysporum (isolate V5w2) and the entomopathogenic Beauveria bassiana (isolate WA) have been shown to increase host resistance to various banana pests and diseases. However, there is limited data on how the combined inoculation of these isolates would affect field performance of bananas. In this study, the fungal endophytes were inoculated separately and in combination. Tissue cultured plantlets of cooking banana cultivar Mbwazirume and dessert banana cultivar Grande Naine were inoculated by root drenching with a suspension of 1.0 × 107 spores mL−1 of the endophytes on three occasions, separated 4 weeks apart, before transplanting into the field. Each plantlet was further inoculated with 1800 nematodes, composed primarily of R. similis. Inoculation of banana plants with the fungal endophytes significantly reduced nematode densities by >34%. Similarly, plant toppling was lower in the endophyte-enhanced plants (<16.5%) compared with the control (23.3%). We also observed improved yield of the first crop cycle in the endophyte-enhanced plants, which yielded >11 t ha−1 year−1 versus 9 t ha−1 year−1 achieved in the non-inoculated plants. These findings demonstrate the benefits of fungal endophytes in improving the yield of both cooking and dessert bananas via suppression of nematode densities and nematode-related damage.

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Abstract

The success of Phasmarhabditis hermaphrodita (Schneider) Andrássy (Rhabditida: Rhabditidae) as a biological control agent of molluscs has led to a worldwide interest in phasmarhabditids. However, scant information is available on the lifecycle development of species within the genus. In the current study, the development of P. hermaphrodita, Phasmarhabditis papillosa, Phasmarhabditis bohemica and Phasmarhabditis kenyaensis were studied using ex vivo cultures, in order to improve our understanding of their biology. Infective juveniles (IJs) of each species were added to 1 g of defrosted homogenized slug cadavers of Deroceras invadens and the development monitored after inoculated IJ recovery, over a period of eight–ten days. The results demonstrated that P. bohemica had the shortest development cycle and that it was able to produce first-generation IJs after eight days, while P. hermaphrodita, P. papillosa and P. kenyaensis took ten days to form a new cohort of IJs. However, from the perspective of mass rearing, P. hermaphrodita has an advantage over the other species in that it is capable of forming self-fertilizing hermaphrodites, whereas both males and females are required for the reproduction of P. papillosa, P. bohemica and P. kenyaensis. The results of the study contribute to the knowledge of the biology of the genus and will help to establish the in vitro liquid cultures of different species of the genus.

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Abstract

The success of the mollusc-parasitic nematode, Phasmarhabditis hermaphrodita (Schneider) Andrássy (Rhabditida: Rhabditidae), as a biological control agent in Europe has led to worldwide interest in phasmarhabditids as biocontrol agents. In this study, the mass culture potential of three phasmarhabditids, namely Phasmarhabditis papillosa, Phasmarhabditis kenyaensis and Phasmarhabditis bohemica, was assessed. In addition, ten bacterial candidates, consisting of seven associated with slugs and three associated with entomopathogenic nematodes, were investigated. The bacteria were tested for their ability to cause mortality to Deroceras invadens, as well as to support nematode growth. Initial mortality studies demonstrated that Kluyvera, Aeromonas and Pseudomonas spp. (AP3) caused 100% mortality when they were injected into the haemocoel of D. invadens. However, in growth studies, Pseudomonas sp. (AP4) was found to be the most successful bacterium, leading to recovery and reproduction in almost all nematode species, except for P. kenyaensis. In flask studies, P. bohemica, which showed exceptional growth with Pseudomonas sp. (AP1), was chosen for further investigation. The effect of inoculating flasks with different concentrations of Pseudomonas sp. (AP1), as well as with different concentrations of P. bohemica, was evaluated by assessing the nematode populations for 14 days. The results indicated that the lowest, 1% (v/v), bacteria inoculation led to higher total nematode and to infective juvenile (IJ) yield, with flasks with the highest IJ inoculum (3000 IJs/ml) having a positive effect on the total number of nematodes and IJs in cultures of P. bohemica. This study presents improvements for the mass-culturing of nematodes associated with molluscs.

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Abstract

Despite their important ecological roles for soil health and soil fertility, free-living nematodes (FLN) have received relatively limited research attention. The present study evaluated the community structure and diversity of FLN in a field setting. The experiments were conducted in on-farm and on-station field plots sown to maize (Zea mays) and beans (Phaseolus vulgaris) under four cropping practices. These farming systems included organic (compost and biopesticide use), conventional (synthetic fertilizer and pesticide applications), farmer practice (organic and synthetic amendments) and a control (non-amended plots). Nineteen genera of free living nematodes, belonging to bacterivores, fungivores, omnivores and predators were recorded. Among these, bacterivores (Cephalobidae and Rhabditidae) were the most dominant group in the organic systems when compared to the conventional and control systems. Farming systems influenced the abundance and diversity of free living nematodes, with the organic farming system having higher values of maturity, enrichment and structural indices than other farming systems. This would indicate greater stability in soil health and improved soil fertility. This implies that the organic farming systems play a key role in improving the biodiversity and population buildup of FLN, compared with other systems. Our study helps to improve our understanding of how farming systems influence soil biodynamics, while studies on the longer-term effects of organic and conventional farming systems on the build-up or reduction of free living nematodes for improved ecosystem services are needed.

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Abstract

Bio-communication occurs when living organisms interact with each other, facilitated by the exchange of signals including visual, auditory, tactile and chemical. The most common form of bio-communication between organisms is mediated by chemical signals, commonly referred to as ‘semiochemicals’, and it involves an emitter releasing the chemical signal that is detected by a receiver leading to a phenotypic response in the latter organism. The quality and quantity of the chemical signal released may be influenced by abiotic and biotic factors. Bio-communication has been reported to occur in both above- and below-ground interactions and it can be exploited for the management of pests, such as cyst nematodes, which are pervasive soil-borne pests that cause significant crop production losses worldwide. Cyst nematode hatching and successful infection of hosts are biological processes that are largely influenced by semiochemicals including hatching stimulators, hatching inhibitors, attractants and repellents. These semiochemicals can be used to disrupt interactions between host plants and cyst nematodes. Advances in RNAi techniques such as host-induced gene silencing to interfere with cyst nematode hatching and host location can also be exploited for development of synthetic resistant host cultivars.

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Abstract

Plant-parasitic nematodes (PPN) cause significant yield reduction in commercial pineapple (Ananas comosus) worldwide. In Kenya, few nematode studies have been conducted, although the main commercial pineapple producer has sole dispensation to use Telone II (1,3-Dichloropropene) indicating the magnitude of the nematode problem. This study was conducted with the aim to investigate the population densities and diversity of nematodes in two commercial plantations with two contrasting management practices. We additionally assessed the influence of crop age and compared this with nearby smallholder pineapple production systems. Soil and root samples were collected from fields of different ages in each commercial plantation and from 29 smallholder fields. A total of 18 genera were associated with pineapple, with a relatively greater diversity found in smallholder than commercial farms. The most prevalent genus was Meloidogyne spp. (M. javanica) followed by Helicotylenchus spp., Tylenchus spp. and Aphelenchoides spp. PPN densities were higher in relatively older fields of 24 and 36 months than from fallow and 3-month-old fields. Regression analysis additionally demonstrated the rise of PPN densities with age of pineapple fields, especially Meloidogyne spp., while free-living nematode densities declined. This study provides an indication of the high level of PPN infection in pineapple in Kenya, which would constitute an important factor contributing to low yields. The study confirms an obvious need for pineapple producers to control PPN to improve crop yields.

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Abstract

The banana weevil (BW), Cosmopolites sordidus, is the main coleopteran pest of banana, causing up to 100% yield loss. In this study, we screened 20 isolates of entomopathogenic fungi (EPF) for the management of BW. In the lab, eight Beauveria bassiana isolates caused >50% mortality of the adult BW, whereas Metarhizium anisopliae and Isaria fumosorosea isolates were less pathogenic. B. bassiana isolates ICIPE 648, ICIPE 660 and ICIPE 273 were the most pathogenic, killing ≥80% of adult BW. B. bassiana isolate ICIPE 622 yielded the highest spores per BW cadaver (1.84 × 108 spores), followed by ICIPE 660, ICIPE 273 and ICIPE 648—1.17 × 108, 3.8 × 107 and 3.6 × 107 spores, respectively. ICIPE 273 had the shortest LT50 (5.3 days) followed by ICIPE 648 (9.8 days) and 660 (11.1 days). Similarly, the LC50 values for the three isolates were 5.18 × 107, 5.49 × 107 and 5.2 × 107 spores mL−1, respectively. In the field, ICIPE 273 and ICIPE 648 had the highest (31.3%) and lowest (20.8%) pathogenicity, respectively. This study indicates that the B. bassiana isolates ICIPE 273, ICIPE 648 and ICIPE 660 are potential candidates for the environmentally sustainable management of BW.

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Abstract

A new species of Phasmarhabditis was isolated from the slug, Polytoxon robustum, from Nairobi, Kenya. The nematode was identified using morphological, morphometric, molecular and phylogenetic analyses. Phasmarhabditis kenyaensis n. sp. is characterised by an infective juvenile with the longest body length in the genus, measuring 1232 (1107-1336) μm, by the presence of males with a bursa bearing nine bilateral pairs of genital bursal papillae and one pair of papilliform phasmids flanking the tail, cephalate paired spicules, with an arc length of 71 (57-81) μm, as well as by females with a vulva located at the mid-body region and a conoid tail shape, with two phasmids located at ca 40% of the tail length. The molecular phylogeny of the new species, as inferred from its SSU (small subunit) rRNA gene, places P. kenyaensis n. sp. genetically close to undescribed phasmarhabditids from South Africa, suggesting an African grouping, while the D2-D3 (large ribosomal subunit) and ITS region analyses relate P. kenyaensis n. sp. to P. meridionalis, with weak bootstrap support. This is the third new Phasmarhabditis species described from the African continent, the new species bringing the total known complement of the genus to 14 species. A morphometric compendium to all species cultured in vivo is supplied.

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Abstract

Enset (Ensete ventricosum), is a perennial herbaceous plant belonging to the family Musaceae, along with banana and plantain. Despite wild populations occurring in eastern, central and southern Africa, it is only in Ethiopia that the crop has been domesticated, where it is culturally and agriculturally symbolic as a food security crop. Although an under-researched orphan crop, enset serves as a staple food for about 20% of the Ethiopian population, comprising more than 20 million people, demonstrating its value in the country. Similar to banana and plantain, enset is heavily affected by plant-parasitic nematodes, with recent studies indicating record levels of infection by the root lesion nematode Pratylenchus goodeyi. Enset is propagated vegetatively using suckers that are purposely initiated from the mother corm. However, while banana and plantain suckers have proven to be a key source of nematode infection and spread, knowledge on the infection levels and role of enset suckers in nematode dissemination is lacking. Given the high levels of plant-parasitic nematodes reported in previous surveys, it is therefore speculated that planting material may act as a key source of nematode dissemination. To address this lack of information, we assessed enset planting material in four key enset growing zones in Ethiopia. A total of 340 enset sucker samples were collected from farmers and markets and analyzed for the presence of nematodes. Nematodes were extracted using a modified Baermann method over a period of 48 h. The root lesion nematode P. goodeyi was present in 100% of the samples, at various levels of infection. These conclusive results show that planting material is indeed a key source of nematode infection in enset, hence measures taken to ensure clean suckers for planting will certainly mitigate nematode infection and spread. The effect of nematode infection on yield and quality on enset remains to be investigated and would be a way forward to complement the nematode/disease studies conducted so far and add valuable knowledge to the current poorly known impact of pests and diseases.

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Abstract

Pratylenchus goodeyi appears to be the most prevalent nematode pest of enset in Ethiopia, where it can occur in extremely high densities. However, the damage to yield or how different enset cultivars react to the nematode has yet to be determined. The current study therefore sought to establish a first assessment of these reactions by enset to P. goodeyi infection. Determining pest resistant cultivars is an important task in developing management strategies. Our study evaluated nine enset cultivars to establish host response and identify potential sources of resistance. In addition, the pathogenicity of P. goodeyi was assessed on three enset cultivars. After 9 months’ growth, significant differences in final population densities (Pf) and reproduction factor (RF) were observed amongst the nine cultivars, with ‘Gefetanuwa’ the most susceptible (Pf = 25 799 and RF = 12.9), and similarly in a repeat experiment for 4.5 months (Pf = 126 534 and RF = 63.3). ‘Maziya’ and ‘Heila’ were the most resistant in the first experiment (Pf < 455 and RF < 0.2) as well as in the repeat, together with ‘Kellisa’ (Pf < 5255 and RF < 2.6). In the pathogenicity experiment four inoculum densities significantly affected the Pf and RF but not among the three cultivars ‘Maziya’, ‘Arkiya’ and ‘Heila’. This is the first known study to assess genotype reaction to P. goodeyi, which shows that there are significant differences in the reactions of different cultivars and that resistance appears to be present in enset.

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Abstract

A survey of helminths associated with terrestrial slugs focusing on the invasive Arion vulgaris and the native A. ater was conducted on populations from France, Germany, Netherlands, Norway and Poland. In total, 648 terrestrial slugs were collected from 18 sample sites, and identified by means of morphological examination, dissection of genitalia and molecular analysis using mitochondrial DNA. In addition to A. vulgaris and A. ater, also A. vulgaris/A. rufus hybrids and A. ater/A. rufus hybrids were collected. Helminth species were identified based on morphological features and sequencing of the 18S and ITS rDNA regions. The parasites included four nematode species: Alloionema appendiculatum, Angiostoma sp., Phasmarhabditis hermaphrodita, Entomelas sp., two trematode species: Brachylaima mesostoma, Eurytrema sp., and one cestode (tapeworm) species: Skrjabinia sp. Alloionema appendiculatum was the most common helminth in the investigated slug populations. Furthermore, we found higher prevalence of trematodes in the invasive A. vulgaris compared with the native A. ater, while differences in the prevalence for nematodes were not as clear.

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Abstract

Enset (Ensete ventricosum) is an important starch staple crop, cultivated primarily in south and southwestern Ethiopia. Enset is the main crop of a sustainable indigenous African system that ensures food security in a country that is food deficient. Related to the banana family, enset is similarly affected by plant-parasitic nematodes. Plant-parasitic nematodes impose a huge constraint on agriculture. The distribution, population density and incidence of plant-parasitic nematodes of enset was determined during August 2018. A total of 308 fields were sampled from major enset-growing zones of Ethiopia. Eleven plant-parasitic nematode taxa were identified, with Pratylenchus (lesion nematode) being the most prominent genus present with a prominence value of 1460. It was present in each sample, with a highest mean population density per growing zone of 16 050 (10 g root)−1, although densities as high as 25 000 were observed in fields at higher altitudes in Guraghe (2200-3000 m a.s.l.). This lesion nematode is found in abundance in the cooler mountainous regions. Visible damage on the roots and corms was manifested as dark purple lesions. Using a combination of morphometric and molecular data, all populations were identified as P. goodeyi and similar to populations from Kenya, Uganda and Spain (Tenerife). Differences in population densities amongst cultivars indicate possible resistance of enset to P. goodeyi.

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Abstract

Potato cyst nematodes (PCN), such as Globodera rostochiensis and Globodera pallida, are quarantine restricted pests of potato causing major yield and financial losses to farmers. G. rostochiensis was first reported from Kenya’s key potato growing area in 2015. We sought to determine the diversity, prevalence and distribution of PCN species across the country by conducting a country-wide survey between 2016 and 2018, which included a more focused, follow-up assessment in three key potato growing counties. A total of 1,348 soil samples were collected from 20 potato growing counties. Information regarding local potato farming practices, potato cultivar use, their diversity and availability was also recorded. PCN cysts were obtained from 968 samples (71.8%) in all the counties surveyed, with Nyandarua County recording the highest PCN field-incidence at 47.6%. The majority of PCN populations, 99.9%, were identified as G. rostochiensis, while G. pallida was recovered from just one field, in a mixed population with G. rostochiensis. Inconsistencies in PCR amplification efficiency was observed for G. rostochiensis using the recommended EPPO primers, compared with ITS primers AB28/TW81, indicating that this protocol cannot be entirely relied upon to effectively detect PCN. Egg density in Nyandarua County varied between 30.6 and 158.5 viable eggs/g soil, with an average egg viability of 78.9 ± 2.8% (min = 11.6%, max = 99.9%). The PCN-susceptible potato cultivar named Shangi was the most preferred and used by 65% of farmers due to its shorter dormancy and cooking time, while imported cultivars (Destiny, Jelly, Manitou, and Markies) with resistance to G. rostochiensis were used by 7.5% of farmers due to unavailability and/or limited access to seeds. Thus, most farmers preferred using their own farm-saved seeds as opposed to purchasing certified seeds. Establishing the distribution and prevalence of PCN and elucidating the local farming practices that could promote the spread of PCN is a necessary precursor to the implementation of any containment or management strategy in the country and ultimately across the region.

Abstract

The invasive slug Arion vulgaris (Gastropoda: Arionidae) is an agricultural pest and serious nuisance in gardens of Central and Northern Europe. To investigate if the success of A.vulgaris in Norway can be attributed to a release from parasites, we compared the prevalence and parasite load of nematodes and trematodes in A. vulgaris to that of three native gastropod species, A. circumscriptus, A. fasciatus and Arianta arbustorum, in SE Norway. We found A. vulgaris to have the highest prevalence of both parasite groups (49% nematodes, 76% trematodes), which does not support the parasite release hypothesis, but rather points to A. vulgaris as a potentially important intermediate host of these parasites. For trematodes the number of individuals (parasite load) did not differ among host species; for nematodes it was higher in A. vulgaris than A. fasciatus. To further compare the parasite susceptibility of the surveyed gastropods, we exposed A. vulgaris, A. fasciatus, and A. arbustorum to a slug parasitic nematode, Phasmarhabditis hermaphrodita, in the laboratory. This nematode is commercially available and widely used to control A. vulgaris. The non-target species A. fasciatus was most affected, with 100% infection, 60% mortality and significant feeding inhibition. A. vulgaris was also 100% infected, but suffered only 20% mortality and little feeding inhibition. The load of P. hermaphrodita in infected specimens was not significantly different for the two Arion species (median: 22.5 and 45, respectively). Only 35% of A. arbustorum snails were infected, none died, and parasite load was very low (median: 2). However, they showed a near complete feeding inhibition at highest nematode dose, and avoided nematode-infested soil. Our results indicate that A. vulgaris may be less susceptible to P. hermaphrodita than the native A. fasciatus, and that non-target effects of applying this nematode in fields and gardens should be further investigated.

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Abstract

Angiostoma norvegicum n. sp. (Angiostomatidae) is described from the oesophagus, crop and the buccal mass of five species of slugs of the family Arionidae, Arion vulgaris (Moquin-Tandon), Arion ater (L.), Arion fasciatus (Nilsson), Arion fuscus (Müller) and Arion rufus/Arion ater hybrid), collected throughout Norway. Angiostoma norvegicum n. sp. was found parasitising arionids at seven of the 30 sample sites examined (23.3%), and 9.9% of all Arion spp. were infected with this nematode. The new species is characterised by its large size (4.0–8.6 mm long) and in having: lateral alae; 6 + 6 papillae at the cephalic end; a large circular mouth aperture; a spacious stoma; a pharyngeal basal bulb without valvular apparatus; an excretory pore near the base of bulb; a distal part of posterior ovary always outstretched; an anterior ovary distally nearly always outstretched; a vulva situated anterior to mid-body; long, nearly straight spicules and a small gubernaculum; three circumcloacal papillae and caudal genital papillae (GP) arranged in a pattern 1+2/3+3 with GP 5 and GP 8 opened on dorsal side of narrow bursa not reaching tail tip; short conical tails in both sexes with tips supplied by 4 short, unequal denticles. Morphologically, A. norvegicum n. sp. is similar to Angiostoma limacis Dujardin, 1845, which diagnostic characteristics are given based on examination of specimens from Norway and the UK. Conversely, the phylogenetic analyses based on D2D3 large subunit (LSU) rRNA gene sequences performed in the present study did not support the morphological affinity of these two species. Phylogenetic analyses demonstrated that although Angiostoma spp. cluster together, A. norvegicum n. sp. forms a tight monophyletic clade with the milacid nematode parasites Angiostoma margaretae Ross, Malan & Ivanova, 2011 and Angiostoma milacis Ivanova & Wilson, 2009.

Abstract

A survey of nematodes associated with terrestrial slugs was conducted for the first time in Norway. A total of 611 terrestrial slugs were collected from 32 sample sites. Slugs were identified by means of morphological examination, dissection of genitalia and molecular analysis using mitochondrial DNA. Twelve slug species were identified, representing four different slug families. Internal nematodes were identified by means of morphological analysis and the sequencing of the 18S rRNA gene. Of the sample sites studied, 62.5% were found to be positive for nematode parasites, with 18.7% of all slugs discovered being infected. Five nematode species were identified in this study: Alloionema appendiculatum, Agfa flexilis, Angiostoma limacis, Angiostoma sp. and Phasmarhabditis hermaphrodita. Of these species, only one nematode was previously undescribed (Angiostoma sp.). This is the first record of the presence of A. appendiculatum, A. flexilis and A. limacis in Norway.

Abstract

The detection in 1999 of the pine wood nematode (PWN), Bursaphelenchus xylophilus, in Portugal triggered survey activities in many European countries. With the assumption that PWN would reach frequency 10 times lower than the native B. mucronatus and the requirement of a 95 % confidence limit suggested 2 995 samples to be required for a safe statement on the absence of PWN from the territory surveyed. Samples were taken from 10 circular areas with 50 km diameter erected from a point of import of risk materials. In the period 2000-2006 3 165 wood samples, 2 880 from Pinus sylvestris, 279 from Picea abies and 6 from unknown wood, were collected from 446 logging sites, in 84 municipalities and 13 counties. Of the total material 85 % of the samples came from cutting wastes, timber or lying trees. Wood showing signs of insect activity (incl. Monochamus) formed 73 % of the total material. Nematodes were recorded in 85 % of the samples. The order Rhabditida was most frequent, followed by the orders Aphelenchida, Tylenchida and Dorylaimida. Rhabditid nematodes were equally frequent in pine and spruce, while Aphelenchida (Aphelenchus, Aphelenchoides, Cryptaphelenchus, Seinura and Bursaphelenchus) and Tylenchida (Filenchus, Lelenchus, Ditylenchus, Deladenus and Nothotylenchus) tended to be more common in pinewood. Aphelenchoides was the most common genus. The genus Bursaphelenchus occurred in 1 % of the samples. B. mucronatus was detected in 0,3 % of the samples and most often in cutting waste of pine. The pine wood nematode (PWN), B. xylophilus, was not detected in this survey. The unexpected low natural occurrence of B. mucronatus indicates that the number of potential niches for PWN also is lower than expected, and hence it will be necessary to continue this surveillance program to reach 10 000 samples. The present zone sites in central and south-eastern Norway will be supplemented with 1-2 zone sites in southwestern region of the country. In the future these zone sites will function as permanent observation areas. Care will also be taken to collect all samples from detached wood with signs of Monochamus activity.

Abstract

In this survey of 2002, 600 samples were collected from 83 forest blocks in the counties Akershus, Buskerud, Oppland and Østfold. The sampling activity involved 16 municipallities situated mainly within the three zone sites A, B, and C. Samples from Scots pine (Pinus sylvestris) formed 89%, while samples of Norway spruce (Picea abies) made up 10% of the total sample volume. Timber and forest debris were the most common objects sampled. Sixty-five percent of the pine samples and 81% of the spruce samples showed signs of Monochamus activity. Nematodes were common and occurred in 94% of the samples analysed. Thirteen samples of pinewood were positive for the genus Bursaphelenchus. Bursaphelenchus mucronatus was recorded for the third time in Norway, and was detected in forest debris attacked by Monochamus at Bjørdalen in the municipality of Eidsberg in the county of Østfold. The pine wood nematode Bursaphelenchus xylophilus was not detected in this survey.

Abstract

In this survey of 2003, 600 samples were collected from 96 forest blocks in the counties of Aust-Agder and Vest-Agder in southern Norway. The sampling activity involved 19 municipalities situated mainly within the two zone sites D and E close to Kristiansand and Arendal. Samples from Scots pine (Pinus sylvestris) formed 92%, while samples of Norway spruce (Picea abies) made up 8% of the total sample volume. Timber and forest debris was the most common objects sampled. Ninety-eight percent of the samples, regardless of tree species, showed signs of Monochamus activity. Nematodes were common and occurred in 90% of the samples analysed. Eight samples of pinewood were positive for the genus Bursaphelenchus. This genus did not occur in spruce. Bursaphelenchus mucronatus was detected in 6 samples of forest debris of pine attacked by Monochamus and collected in the county of Aust-Agder. In the municipality of Evje and Hornes B. mucronatus was detected at Skjerkelia and Sutestad. In the municipality of Froland the nematode was found in two samples from Budalsfjellet, and in one sample from Mjålandsvatn. In the municipality of Birkenes one sample from Vågsdalen contained B. mucronatus. This is the fourth report on the occurrence of B. mucronatus in Norway. The pine wood nematode Bursaphelenchus xylophilus was not detected.

Abstract

In 2001, a zone site C, was established as a circular areas with 50 km radius and centred in Greåker close to Sarpsborg in South-eastern Norway. Zone site C is complementary to the similar zone sites A and B established in 2000. From June 2001 to December 2002, 601 wood samples were collected from 66 forest blocks, all situated within the 3 existing zone sites A, B and C. The sampling was carried out in the provinces Akershus, Buskerud, Vestfold and Østfold, but was concentrated to the South-eastern region, where 399 samples were taken from 45 forest blocks in the province of Østfold. Samples from Scots pine (Pinus sylvestris) formed 96% of the collected material, while Norway spruce (Picea abies) was a minor fraction. Lying trees and various kinds of detached wood formed the main part of the objects sampled. The frequency of objects showing signs of attack by wood boring insects was 44%. Some samples were also taken in Porsgrunn in the province of Telemark from a consignment of spruce imported from Russia. Nematodes were often more common in samples from objects with signs of insect activity. In four samples, all from lying pine trees, nematodes belonging to the genus Bursaphelenchus were detected. Bursaphelenchus mucronatus was recorded for the second time in Norway, and occurred in a lying pine tree attacked by Monochamus at Ombudstvedt in the municipality of Våler in the province of Østfold. The pine wood nematode Bursaphelenchus xylophilus was not detected, nor in forests or in 5 samples from spruce imported from Russia

Abstract

Two zone sites, i.e. two circular areas with 50 km radius, were established in southern Norway. The zone sites were centred in Tofte (the location of a major pulp mill) and in Drammen (the site of a major timber yard). From June to October 2000, 66 forest blocks were visited, 65 of which were situated within the zone site areas. Samples were collected from 40 forest blocks, especially from wood attacked by wood boring insects. At 34 forest blocks, trees of Scots pine, Pinus sylvestris, or Norway spruce Picea abies were provided as trap-logs for Monochamus spp. This material will be sampled in the survey of 2001. Some samples were also taken from a wood chip pile and from imported wood material. The total number of wood samples analysed for nematodes were 275. Out of these, 214 samples were collected from forest trees, stumps, timber and logging wastes of P. sylvestris and P. abies. Three samples contained nematodes belonging to the genus Bursaphelenchus, but the Pine Wood Nematode (PWN), B. xylophilus, was not detected. Similarly, this nematode was not detected in the 10 samples of wood chips, or in the 25 samples of imported lumber or in the 26 samples of imported solid wood packing material. In order reach the minimal number of 3000 samples within reasonable time, the number of samples for the next survey season of 2001 needs to be increased drastically. To achieve this, the sampling will continue within the existing zone sites, and be extended into new zone sites to be established in 2001.