Hanne Steel

Mononchoides composticola n. sp. (Nematoda: Diplogastridae) associated with composting processes: morphological, molecular and autecological characterisation

Mononchoides composticola n. sp. was isolated from compost and is described based on light and scanning electron microscopy, supplemented with SSU rDNA sequence data. It is characterised by the following features: a denticulate ridge in addition to the dorsal claw-like tooth, a small tooth-like swelling at the stegostom base, ca 26 longitudinal ridges on the female body, a uterine sac associated with two dumb-bell-shaped pouches, relatively small spicules (30-38 µm long), a simple gubernaculum shorter than half the spicule length, the genital subventral papillae (v6) consisting of three very small papillae, and a long filiform tail (female: 391-550 µm, 18-26 anal body diam.; male: 304-548 µm, 19-30 anal body diam.). Phylogenetic analyses placed the new species together with M. striatus, sister to Tylopharynx foetida. Since the use of nematodes as functional indicators often relies on the allocation of nematodes to feeding groups, experiments were performed to elucidate the feeding strategy of the new species. Both its ability to move actively to bacterial food sources and to prey on other compost nematodes were tested. Mononchoides composticola n. sp. actively moved towards the compost bacterium Achromobacter, a taxis that was temperature dependent, and also preyed on other nematodes. Predation was selective, with a higher predation rate on the relatively small and slow-moving Rhabditella sp. than on the considerably larger and more motile Rhabditis (Poikilolaimus) sp. Adults of M. composticola n. sp. have a dual feeding behaviour and can apparently alternate between bacterial and nematode prey

Beyond plant–soil feedbacks: mechanisms driving plant community shifts due to land‐use legacies in post‐agricultural forests

Summary Although biotic legacies of past agricultural practices are widespread and increasing in contemporary ecosystems, our understanding of the mechanisms driving such legacies is still poor. Forest understories on former agricultural land show low frequencies and abundance of typical woodland species when compared with ancient forests. These community shifts have been ascribed to the effects of dispersal limitation. A rarely considered mechanism is that post-dispersal processes driven by plant-associated communities determine the poor performance and recruitment of woodland indicators. Given the strong alterations in soil conditions due to former agricultural practices, we hypothesized that (abiotic) plant–soil feedbacks could be a major factor in community shifts. We addressed this hypothesis by comparing plant-associated communities in the soil and above the ground in ancient and post-agricultural alluvial forests; then, we experimentally tested whether the changes in biotic and abiotic soil properties could affect above-ground herbivore abundance and pressure and plant performance. Ancient and post-agricultural communities clearly differed in composition at different levels of the food web. Besides the plant community, we also observed the differences in the microbial and nematode community with increased abundance of root-feeding nematodes in post-agricultural soils. The composition of the above-ground invertebrate community did not differ in ancient and post-agricultural forest parcels; however, plants growing in post-agricultural sites showed higher abundance of invertebrate herbivores and suffered more herbivory. Nutrient analyses of soil and plants showed that increased levels of phosphorus (and to a lesser extent, nitrogen) made plants more nutritious for insect herbivores. Laboratory experiments further pointed to this mechanism as an explanation of the poorer performance of woodland indicators in post-agricultural woodlands. Our results point to biotic and abiotic plant–soil feedbacks coupled with herbivory as a new mechanism to explain the legacy effects in temperate forests. The modification of the below-ground community and soil abiotic characteristics by previous agricultural activity affects not only the plant growth but also the plant nutrient content in the compared understorey species, making them more susceptible to above-ground herbivory. Our results provide one of the first examples of integrating plant–soil feedback and above- and below-ground interactions to explain land-use legacies.

Nematodes enhance plant growth and nutrient uptake under C and N-rich conditions

The role of soil fauna in crucial ecosystem services such as nutrient cycling remains poorly quantified, mainly because of the overly reductionistic approach adopted in most experimental studies. Given that increasing nitrogen inputs in various ecosystems influence the structure and functioning of soil microbes and the activity of fauna, we aimed to quantify the role of the entire soil nematode community in nutrient mineralization in an experimental set-up emulating nutrient-rich field conditions and accounting for crucial interactions amongst the soil microbial communities and plants. To this end, we reconstructed a complex soil foodweb in mesocosms that comprised largely undisturbed native microflora and the entire nematode community added into defaunated soil, planted with Lolium perenne as a model plant, and amended with fresh grass-clover residues. We determined N and P availability and plant uptake, plant biomass and abundance and structure of the microbial and nematode communities during a three-month incubation. The presence of nematodes significantly increased plant biomass production (+9%), net N (+25%) and net P (+23%) availability compared to their absence, demonstrating that nematodes link below- and above-ground processes, primarily through increasing nutrient availability. The experimental set-up presented allows to realistically quantify the crucial ecosystem services provided by the soil biota.

Experimental induction of intraspecific morphometric variability in a single population of Halicephalobus cf. gingivalis may surpass total interspecific variability

Although molecular techniques are revolutionising nematode taxonomy, morphological data still form the basis of nematode species descriptions. However, morphological characters show a natural variability that should be taken into account before describing new species. The current study presents the results of an elaborate morphometric study of Halicephalobus cf. gingivalis, including 15 measurements and 13 indices of 540 specimens, the progeny of a single parthenogenetic female and cultured under different temperature and food conditions and measured in different adult age groups, i.e., young adults with a developed vulva but before the onset of oviposition, adults laying eggs, and old, post-reproductive adults near the end of their life cycle. The morphometric characters were analysed using both univariate (analysis of variance) and multivariate (principal components and canonical discriminant analysis) techniques. The main results reveal that the morphometric characters most used in Halicephalobus identification keys have a huge variability within a single progeny, e.g., body length 1.9 times longer than the shortest or ratio VA/tail length 3.9 times larger than the smallest. This variability has a magnitude that has not been observed in nematodes before. Further, by changing the environmental factors, the morphometric characters are influenced to an extent that one could assign – with seemingly ‘statistical support’ – different ‘species’ of the genus to different subpopulations. With this experimental study we provide convincing elements to advocate an integrative taxonomic approach and to discourage the description of new species based only on morphometric differences.

Intestine ultrastructure of the facultative parasite Halicephalobus gingivalis (Nematoda: Panagrolaimidae)

Halicephalobus gingivalis, classified as a free-living nematode, is a known facultative parasite of horses, zebras and humans. However, detailed information concerning its internal morphology is lacking, especially in relation to possible adaptations to its lifestyle as a facultative parasite. The research presented here uses TEM and PI staining to analyse the morphology of the intestine of H. gingivalis. Specimens cultured under different conditions were included to determine if differences in ultrastructure are induced by culturing method. TEM analysis revealed that the intestinal tract comprises a single layer of cells in which nine pairs of nuclei can be distinguished. Further, unusual dichotomously and trichotomously branched microvilli were observed next to finger-like cylindrical microvilli, the latter being the most commonly described form in nematodes. Finally, three different types of secretion vesicles, i.e., spherical (type I), thread-like (type II) and enlarged globular (type III), occurred independently from each other along the intestinal tract. The relationship of morphological adaptations of the microvilli to parasitism in nematodes is discussed.

Quantifying the Contribution of Entire Free-Living Nematode Communities to Carbon Mineralization under Contrasting C and N Availability

To understand the roles of nematodes in organic matter (OM) decomposition, experimental setups should include the entire nematode community, the native soil microflora, and their food sources. Yet, published studies are often based on either simplified experimental setups, using only a few selected species of nematode and their respective prey, despite the multitude of species present in natural soil, or on indirect estimation of the mineralization process using O2 consumption and the fresh weight of nematodes. We set up a six-month incubation experiment to quantify the contribution of the entire free living nematode community to carbon (C) mineralization under realistic conditions. The following treatments were compared with and without grass-clover amendment: defaunated soil reinoculated with the entire free living nematode communities (+Nem) and defaunated soil that was not reinoculated (-Nem). We also included untreated fresh soil as a control (CTR). Nematode abundances and diversity in +Nem was comparable to the CTR showing the success of the reinoculation. No significant differences in C mineralization were found between +Nem and -Nem treatments of the amended and unamended samples at the end of incubation. Other related parameters such as microbial biomass C and enzymatic activities did not show significant differences between +Nem and -Nem treatments in both amended and unamended samples. These findings show that the collective contribution of the entire nematode community to C mineralization is small. Previous reports in literature based on simplified experimental setups and indirect estimations are contrasting with the findings of the current study and further investigations are needed to elucidate the extent and the mechanisms of nematode involvement in C mineralization.

Deceptive morphological variation in Hirschmanniella mucronata (Nematoda: Pratylenchidae) and a polytomous key to the genus

Hirschmanniella mucronata populations isolated from two Cambodian provinces were characterised using morphological, morphometric and molecular criteria. Examination of 1024 specimens from 60 different paddy fields revealed high intraspecific variation in morphology and morphometrics, especially in tail terminus shape and stylet length. Sequence results confirmed that morphologically divergent individuals represent a single species, suggesting that neglecting morphological variation has led to an overestimation of Hirschmanniella diversity in former studies. Phylogenetic analysis of the SSU, D2-D3, LSU and ITS1-5.8S-ITS2 regions revealed three concordant clades, H. mucronata having a sister relationship with H. kwazuna and H. loofi. Plotting the diagnostic features, including tail terminus shape, stylet length and lip region morphology on the phylogenetic framework, revealed that none of them supported the clades and represented convergent features. All three molecular markers were able to discriminate all Hirschmanniella species, but the D2-D3 region was the easiest, fastest and most successful region to be amplified. Species delimitation and the diagnostic features of Hirschmanniella were re-evaluated. Hirschmanniella abnormalis and H. exacta are considered to be junior synonyms of H. oryzae and H. mannai a species inquirenda. Finally, a list of valid species with indication of synonyms and a polytomous key are provided.

Soil and litter nematode diversity of Mount Hamiguitan, the Philippines, with description of Bicirronema hamiguitanense n. sp. (Rhabditida: Bicirronematidae)

The nematode diversity in soil and litter was investigated on Mount Hamiguitan, the Philippines, along four eco-habitats from elevations of 75-1600 m a.s.l. A total of 155 and 467 nematodes were identified to 39 and 62 genera from litter and soil, respectively. The nematode assemblages and diversity did not show any relation to eco-habitat or elevation. Bacterivorous nematodes were the most common group (37.5%). Acrobeloides was most abundant from the soil and Aphelenchoides from the litter. Bicirronema hamiguitanense n. sp. is herein described based on morphology, morphometrics and molecular data. The new species has the following diagnostic features: a wide lateral field one-fifth of its body diam. with four incisures forming two ornamented ridges; gubernaculum with proximal thickening; spicules (35-38 mu m) and gubernaculum (18-20 mu m) longer than B. caledoniense; and with 37 molecular autapomorphies supporting its new species status. The phylogenetic position of the new species within Cephalobomorpha is discussed.

Nematodes from terrestrial, freshwater and brackish water habitats in Belgium: an updated list with special emphasis on compost nematodes

A study of nematodes from a semi-artificial and controlled composting process in Eastern Flanders revealed 35 taxa, 21 of which were new records for Belgium. An updated checklist of free-living, plant-parasitic and entomopathogenic nematodes from terrestrial, freshwater and brackish water habitats in Belgium is presented. The Belgian non-marine nematofauna comprises 418 taxa, representing 4 subclasses, 14 orders, and 76 families. In total 127 new records were added: i.e. 21 from the newly explored compost habitat, 7 from freshwater samples and 99 from published data in literature.