Wilfrida Decraemer

Effectiveness of different Heterorhabditis isolates from Southern Benin for biocontrol of the subterranean termite, Macrotermes bellicosus (Isoptera: Macrotermitinae), in laboratory trials

The host-finding ability of 28 Heterorhabditis sonorensis isolates and one H. indica isolate from southern Benin and one H. indica isolate from India was examined in vertical migration sand bioassays against workers of a target citrus termite pest, Macrotermes bellicosus. Thereafter, nine selected isolates were subjected to further investigations on virulence. Our results revealed that both H. sonorensis and H. indica isolates exhibit a cruiser type of search strategy and were capable, to various degrees, of migrating, infecting and killing workers of M. bellicosus in sand columns up to 20 cm long over a period of 3 days. However, only three isolates of H. sonorensis caused 100% mortality to M. bellicosus at the greatest depth tested. The exotic H. indica isolate (LN2) did not show strong finding ability compared to the indigenous one. Concerning virulence, differences were observed among isolates for their ability to invade workers of M. bellicosus. After 12 h post exposure, H. sonorensis from Ze (Ze2) and H. sonorensis from Azohoue (Azohoue2) exhibited the lowest invasion time with IT50 = 3.35 and 3.67 h, respectively, and a higher penetration rate (11.4% and 10%, respectively) compared with the other isolates. In the concentration-mortality test, we found that, based on 95% confidence limits, all H. sonorensis and H. indica isolates appeared to be equal, with LC50 values ranging from nine to 16 infected juveniles (IJ) termite I. Interestingly, 40 II termite(-1) were enough to cause 80% mortality to M. bellicosus. Contrary to the LC50, the results of our studies clearly demonstrate that M. bellicosus exhibits a time-dependent susceptibility to the tested nematode isolates. So, the lowest LT50 was observed for H. sonorensis Ze2 (LT50 = 23.30 h), the highest for H. sonorensis Yokon (34.76 h). The LT50 of the indigenous H. indica isolate was estimated to 24.07 h. In addition, all selected isolates were able to reproduce in M. bellicosus workers. The highest reproduction potential in M. bellicosus was observed with H. sonorensis Yokon with 20 213 IJ/termite followed by H. sonorensis Ze2 with 19 368 IJ/termites. All tested Beninese EPN isolates were pathogenic to the citrus termite pest M. bellicosus, with H. sonorensis Ze2 being the most virulent.

Molecular phylogeny of Malenchus and Filenchus (Nematoda: Tylenchidae)

The family Tylenchidae is phylogenetically important to understanding early‐branching Tylenchomorpha and to assess soil ecosystems. In this study, we focus on Malenchus and Filenchus as representatives of the Tylenchidae. Samples collected worldwide result in 58 new sequences, and light microscopy and transmission electron microscopy provide details on morphological features. For the first time, comprehensive morphological data are evaluated in the context of a molecular framework, thus highlighting the phylogenetic and evolutionary complexity of this structurally minimalistic group. Results show that the genus Filenchus is polyphyletic in both the 18S and 28S rDNA phylogeny, while Malenchus is polyphyletic and monophyletic in the 28S rDNA and the 18S rDNA, respectively. Ultrastructural study demonstrates specific aspects of lateral cuticular incisures, cuticular layering and the amphideal fovea are surprisingly congruent with the obtained molecular phylogenies, while classical characteristics such as cuticle annulations are evolutionary highly plastic and mosaic in distribution. The study also reveals the shortage of D2/D3 domain in 28S rDNA as a phylogenetic marker for early‐branching Tylenchomorpha.

Influence of pesticides, soil temperature and moisture on entomopathogenic nematodes from southern Benin and control of underground termite nest populations

The influence of three pesticides on the viability and infectivity of four Beninese isolates of entomopathogenic nematodes (EPN), Heterorhabditis indica Ayogbe1, H. sonorensis Azohoue2, H. sonorensis Ze3, and Steinernema sp. Bembereke, was determined. The impact of both soil temperature and soil moisture on the virulence of these EPN to Trinervitermes occidentalis was investigated in laboratory assays. The effect of EPN-infected Galleria mellonella larvae on underground populations of Macrotermes bellicosus was also examined. All tested Heterorhabditis species were more tolerant to glyphosate and fipronil than the Steinernema species. Heterorhabditis sonorensis Azohoue2, showed the best results with 63.2% termite mortality at a soil temperature of 35°C. The increase of soil moisture to 20% (w/w) did not negatively influence the virulence of tested EPN. The underground populations of 71% or 60% treated nests were controlled by H. sonorensis Azohoue2- or H. indica Ayogbe1-infected G. mellonella larvae, respectively.

Molecular diversity of Photorhabdus and Xenorhabdus bacteria, symbionts of Heterorhabditis and Steinernema nematodes retrieved from soil in Benin

The diversity of 43 bacterial strains isolated from Beninese entomopathogenic nematodes was investigated molecularly by analyzing the 16S rRNA, recA, and gyrB genes. Based on 16S rRNA sequence analysis, 15 bacterial strains were identified as Xenorhabdus sp., 27 strains as Photorhabdus sp., and one as Serratia sp. The Xenorhabdus strains were isolated from Steinernema nematodes and identified as Xenorhabdus indica based on 16S rRNA gene and concatenated recA and gyrB sequence analysis. However, analysis of 16S rRNA and concatenated recA and gyrB gene sequences of the Photorhabdus strains, all isolated from Heterorhabditis nematodes, resulted in two separate sub-clusters (A) and (B) within the Photorhabdus luminescens group, distinct from the existing subspecies. They share low sequence similarities with nearest phylogenetic neighbors Photorhabdus luminescens subsp. luminescens HbT, Photorhabdus luminescens subsp. caribbeanensis HG29T, and Photorhabdus luminescens subsp. noenieputensis AM7T.

Steinernema kandii n. sp. (Rhabditida: Steinernematidae), a new entomopathogenic nematode from northern Benin

Two nematode isolates from the genus Steinernema were collected in northern Benin. Morphological, morphometric, molecular and cross-hybridisation studies placed these nematodes into a new species, Steinernema kandii n. sp., within the bicornutum-group. Phylogenetic analyses based on both ITS and D2-D3 regions of 28S rDNA revealed that S. kandii n. sp. is different from all known Steinernema species and sister to S. abbasi (97.3-97.6% ITS nucleotide similarity) and S. bifurcatum (98.3-98.4% D2-D3 similarity). Steinernema kandii n. sp. can be separated from other members of the bicornutum-group by the greater infective juvenile (IJ) max. body diam. of 35 (27-48) μm (type isolate). It differs from S. abbasi by the greater IJ body length 707 (632-833) μm (type isolate), EP distance 55 (52-60) μm (type isolate), spicule length 67 (57-75) μm (type isolate) and the occurrence of one pair of genital papillae at the cloacal aperture.