Chitin-enriched insect frass fertilizer as a biorational alternative for root-knot nematode (Meloidogyne incognita) management.

Root-knot nematodes (Meloidogyne spp.) are serious pests of most food crops, causing up to 100% yield loss. Nevertheless, commercial nematicides are costly and harmful to the environment. While the nematicidal potential of crustacean and synthetic chitin has been demonstrated globally, research on the potential of insect-derived chitin for nematode control has received limited attention. Here, seven chitin-fortified black soldier fly frass fertilizer extracts (chFE) were assessed for their suppressiveness of Meloidogyne incognita and impacts on spinach growth in comparison with a commercial nematicide using in vitro and in vivo bioassays. The performance of chFE and control treatments was assessed by determining their effects on nematode egg hatchability; infective juvenile (J2) mortality and paralysis; number of galls, egg masses, and J2s per plant; and spinach root and shoot biomass. In vitro results showed that chFE and commercial nematicide suppressed nematode egg hatchability by 42% and 52%, respectively, relative to the control (sterile distilled water). Up to 100% paralysis was achieved when M. incognita J2s were exposed to either chFE or commercial nematicide. Further, the J2 mortality achieved using chFE (95%) was comparable to the value achieved using commercial nematicide (96%); in all treatments, mortality increased with exposure time. Similarly, up to 85% suppression of gall development was achieved when spinach plants were grown in soil drenched with chFE; up to 79% reduction in egg mass formation and 68% suppression of J2 development in the root system were achieved using chFE. Also, chFE application significantly increased spinach root and shoot biomass by 54%-74% and 39%-58%, respectively, compared to commercial nematicide. Our findings demonstrate the nematicidal potential of chFE and its benefits on crop production. Thus, chFE could be considered as a promising multipurpose, regenerative, and cost-effective input for sustainable management of plant-parasitic nematodes and enhancement of crop yield.

Evolving dynamics of insect frass fertilizer for sustainable nematode management and potato production.

Potato production faces major challenges from inadequate soil fertility, and nematode infestation, yet synthetic fertilizers and nematicides are costly and harmful to the environment. This study explored the potential of chitin-fortified black soldier fly-composted organic fertilizer (BSFCOF) as a multipurpose organic fertilizer amendment for enhancing potato yield and suppressing potato cyst nematodes (PCN). The BSFCOF was applied at a rate equivalent to 150 kg N ha-1 and fortified with chitin from black soldier fly pupal exuviae at inclusion rates equivalent to 0.5, 1, 2, 3, 4 and 5% chitin. Data were collected on potato growth characteristics, PCN population densities, and soil chemical properties for two growing cycles. Results showed that chitin fortified BSFCOF significantly improved potato growth parameters, chlorophyll concentration, marketable tuber yield and number of marketable tubers. The marketable tuber yield achieved using chitin-fortified BSFCOF was 70 - 362%, and 69 - 238% higher than the values achieved using unfertilized soil during the first and second growing cycles, respectively. Soil amendment with chitin-fortified BSFCOF significantly reduced the number of cysts per 200 g soil-1, number of eggs and J2 per cyst-1, eggs g-1 soil and reproduction rate by 32 - 87%, 9 - 92%, 31- 98% and 31 - 98%, respectively. The PCN suppression increased with chitin inclusion rates. There were significantly higher values for soil pH, ammonium nitrogen, nitrate nitrogen, available phosphorus, calcium, magnesium, potassium, and cation exchange capacity in soil amended with BSFCOF compared to unamended soil. This study demonstrates that BSFCOF fortified with 5% chitin is an effective soil enhancer with multiple benefits, including improved soil fertility, potato performance, and effective management of potato cyst nematodes.

Biocontrol-based strategies for improving soil health and managing plant-parasitic nematodes in coffee production.

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.

Planting Material of Enset (Ensete ventricosum), a Key Food Security Crop in Southwest Ethiopia, Is a Key Element in the Dissemination of Plant-Parasitic Nematode Infection.

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.

Cyst nematode bio-communication with plants: implications for novel management approaches.

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. © 2020 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Potato Cyst Nematodes: A New Threat to Potato Production in East Africa.

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.

Prevalence and parasite load of nematodes and trematodes in an invasive slug and its susceptibility to a slug parasitic nematode compared to native gastropods.

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.

African Nightshade and African Spinach Decrease Root-Knot Nematode and Potato Cyst Nematode Soil Infestation in Kenya.

Plant-parasitic nematodes, particularly root-knot nematodes (RKN: Meloidogyne spp.) and cyst nematodes (CN: Globodera and Heterodera spp.) cause severe yield reduction in most cultivated crops and are of high economic importance. African nightshade (Solanum spp.) and African spinach (Amaranthus spp.) are important African indigenous vegetables (AIV) and are rich sources of nutrition and income. However, their host status to plant-parasitic nematodes remains largely speculative. Therefore, a survey was conducted which revealed that S. villosum exhibited high root galling, whereas on S. scabrum, A. cruentus, and A. dubius root galling was rare or very low. Additionally, soil collected from the rhizosphere of S. villosum and S. scabrum contained few cysts of potato cyst nematodes (PCN), and no developing PCN females were observed on the roots of growing plants. Therefore, we studied the dynamics of RKN and PCN on A. dubius, A. cruentus, S. scabrum, and S. villosum over 2 years in a field experiment. The effects of AIV crop species on RKN and PCN soil infestation were evaluated using susceptible S. lycopersicum or S. tuberosum. After first, second, and third cultivation of A. dubius, A. cruentus, and S. scabrum, RKN infestation of the soil decreased by more than 85%, whereas S. scabrum and S. villosum decreased PCN densities by more than 80%. When cropping susceptible crops, after three seasons of successive cultivation of these AIV, galling index and number of developing PCN females measured on susceptible crops decreased by more than 75%. Wilting and RKN-PCN coinfection incidences also decreased significantly. Here, we present data that support the development of a novel cropping system including African spinach and African nightshade, which reveals a high potential to manage RKN and PCN in an environmentally friendly, effective, and productive way.

Management of Cyst and Root Knot Nematodes: A Chemical Ecology Perspective.

Plant parasitic nematode infection of crops can be highly detrimental to agricultural production. Since the discovery that plant roots release chemicals that attract the infective stage of plant parasitic nematodes some 80 years ago, significant progress in identifying the signaling molecules has occurred only relatively recently. Here, we review the literature on chemical ecological studies of two major plant parasitic nematode groups: root knot nematodes in the genus Meloidogyne and cyst nematodes in the genus Globodera because of the negative impact their parasitism has on farming systems in Africa. We then highlight perspectives for future directions for their management.

Plant-Parasitic Nematodes and Food Security in Sub-Saharan Africa.

Sub-Saharan Africa (SSA) is a region beset with challenges, not least its ability to feed itself. Low agricultural productivity, exploding populations, and escalating urbanization have led to declining per capita food availability. In order to reverse this trend, crop production systems must intensify, which brings with it an elevated threat from pests and diseases, including plant-parasitic nematodes. A holistic systems approach to pest management recognizes disciplinary integration. However, a critical under-representation of nematology expertise is a pivotal shortcoming, especially given the magnitude of the threat nematodes pose under more intensified systems. With more volatile climates, efficient use of water by healthy root systems is especially crucial. Within SSA, smallholder farming systems dominate the agricultural landscape, where a limited understanding of nematode problems prevails. This review provides a synopsis of current nematode challenges facing SSA and presents the opportunities to overcome current shortcomings, including a means to increase nematology capacity.

Angiostoma norvegicum n. sp. (Nematoda: Angiostomatidae) a parasite of arionid slugs in Norway.

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.

The effects of strawberry cropping practices on the strawberry tortricid (Lepidoptera: Tortricidae), its natural enemies, and the presence of nematodes.

Cropping practice can affect pests and natural enemies. A three-year study of the strawberry tortricid, Acleris comariana (Lienig and Zeller) (Lepidoptera: Tortricidae), its parasitoid Copidosoma aretas Walker (Hymenoptera: Encyrtidae), and its entomopathogenic fungi was conducted in seven pairs of organic and conventional farms to test the hypothesis that farming practice (organic versus conventional) will affect the level of pest infestation and will affect the natural enemies. In addition, the number of years with strawberries on the farm, field age, and other factors that may affect pests and their natural enemies were considered. Farms were characterized by their cropping practices, cropping history, and other parameters. Field-collected larvae were laboratory reared to assess mortality from parasitoids and entomopathogenic fungi. In 2010, a survey of nematodes was made to assess the response of an unrelated taxonomic group to cropping practice. 2,743 larvae were collected. Of those, 2,584 were identified as A. comariana. 579 A. comariana were parasitized by C. aretas and 64 A. comariana were parasitized by other parasitoid species. Finally, 28% of the larvae and pupae of A. comariana died from unknown causes. Only two of the field-collected A. comariana larvae were infected by entomopathogenic fungi; one was infected by Isaria sp. and the other by Beauvaria sp. The density of A. comariana was on average four times lower in organic farms, which was significantly lower than in conventional farms. A. comariana was more dominant on conventional farms than on organic farms. The effect of crop age (One, two, or three years) on A. comariana infestation was significant, with higher infestations in older fields. Crop age had no effect on A. comariana infestation in a comparison of first- and second-year fields in 2010. Cropping practice did not lead to significant differences in the level of total parasitism or in C. aretas parasitism; however, C. aretas contributed to a higher proportion of the parasitized larvae on conventional farms than on organic farms. Mortality from unknown causes of A. comariana was higher in organic farms than conventional farms, and unknown mortality was two to seven times higher in second-generation A. comariana than in first generation. Entomopathogenic nematodes were found on three organic farms and one conventional farm. Plant parasitic nematodes were found in more samples from conventional farms than from organic farms. The low density of A. comariana in organic farms exposes the specialist C. aretas to a higher risk of local extinction. In organic farms, where the density of A. comariana is low, other parasitoids may play an important role in controlling A. comariana by supplementing C. aretas. Other tortricid species may serve as alternative hosts for these other parasitoids, contributing to conserving them in the habitat. The higher unknown mortality of larvae from organic fields may be the result of non-consumptive parasitoid or predator effects. This study reports an example of the effects of cropping practice on an insect pest, with similar effects on nematodes. An understanding of the responsible factors could be used to develop more sustainable cropping systems.

Invasive slug populations (Arion vulgaris) as potential vectors for Clostridium botulinum.

BACKGROUND: Norwegian meadows, including those for silage production, are recently found heavily invaded by the slug Arion vulgaris in exposed areas. As a consequence, large numbers of slugs might contaminate grass silage and cause a possible threat to animal feed quality and safety. It is well known that silage contaminated by mammalian or avian carcasses can lead to severe outbreaks of botulism among livestock. Invertebrates, especially fly-larvae (Diptera), are considered important in the transfer of Clostridium botulinum type C and its toxins among birds in wetlands. C. botulinum form highly resistant spores that could easily be consumed by the slugs during feeding. This study aimed to determine whether Arion vulgaris could hold viable C. botulinum and enrich them, which is essential knowledge for assessing the risk of botulism from slug-contaminated silage. Slug carcasses, slug feces and live slugs were tested by a quantitative real-time PCR (qPCR) method after being fed ≅ 5.8 × 104 CFU C. botulinum type C spores/slug. RESULTS: Low amounts of C. botulinum were detected by qPCR in six of 21 slug carcasses with an even spread throughout the 17 day long experiment. Declining amounts of C. botulinum were excreted in slug feces up to day four after the inoculated feed was given. C. botulinum was only quantified the first two days in the sampling of live slugs. The viability of C. botulinum was confirmed for all three sample types (slug carcasses, slug feces and live slugs) by visible growth in enrichment media combined with obtaining a higher quantification cycle (Cq) value than from the non-enriched samples. CONCLUSIONS: Neither dead nor live invasive Arion vulgaris slugs were shown to enrich Clostridium botulinum containing the neurotoxin type C gene in this study. Slugs excreted viable C. botulinum in their feces up to day four, but in rapidly decreasing numbers. Arion vulgaris appear not to support enrichment of C. botulinum type C.

Spatiotemporal analysis of predation by carabid beetles (Carabidae) on nematode infected and uninfected slugs in the field.

The dynamics of predation on parasites within prey has received relatively little attention despite the profound effects this is likely to have on both prey and parasite numbers and hence on biological control programmes where parasites are employed. The nematode Phasmarhabditis hermaphrodita is a commercially available biological agent against slugs. Predation on these slugs may, at the same time, result in intraguild predation on slug-parasitic nematodes. This study describes, for the first time, predation by carabid beetles on slugs and their nematode parasites on both spatial and temporal scales, using PCR-based methods. The highest nematode infection levels were found in the slugs Deroceras reticulatum and Arion silvaticus. Numbers of infected slugs decreased over time and no infected slugs were found four months after nematode application. The density of the most abundant slug, the invasive Arion vulgaris, was positively related to the activity-density of the carabid beetle, Carabus nemoralis. Predation on slugs was density and size related, with highest predation levels also on A. vulgaris. Predation on A. vulgaris decreased significantly in summer when these slugs were larger than one gram. Predation by C. nemoralis on slugs was opportunistic, without any preferences for specific species. Intraguild predation on the nematodes was low, suggesting that carabid beetles such as C. nemoralis probably do not have a significant impact on the success of biological control using P. hermaphrodita.