Artykuły naukowe (WM)
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- ItemBacteria from the midgut of common cockchafer (Melolontha melolontha L.) larvae exhibiting antagonistic activity against bacterial symbionts of entomopathogenic nematodes: isolation and molecular identification(MDPI, 2020-01-16) Skowronek, Marcin; Sajnaga, Ewa; Pleszczyńska, Małgorzata; Kazimierczak, Waldemar; Lis, Magdalena; Wiater, AdrianThe mechanisms of action of the complex including entomopathogenic nematodes of the genera Steinernema and Heterorhabditis and their mutualistic partners, i.e., bacteria Xenorhabdus and Photorhabdus, have been well explained, and the nematodes have been commercialized as biological control agents against many soil insect pests. However, little is known regarding the nature of the relationships between these bacteria and the gut microbiota of infected insects. In the present study, 900 bacterial isolates that were obtained from the midgut samples of Melolontha melolontha larvae were screened for their antagonistic activity against the selected species of the genera Xenorhabdus and Photorhabdus. Twelve strains exhibited significant antibacterial activity in the applied tests. They were identified based on 16S rRNA and rpoB, rpoD, or recA gene sequences as Pseudomonas chlororaphis, Citrobacter murliniae, Acinetobacter calcoaceticus, Chryseobacterium lathyri, Chryseobacterium sp., Serratia liquefaciens, and Serratia sp. The culture filtrate of the isolate P. chlororaphis MMC3 L3 04 exerted the strongest inhibitory effect on the tested bacteria. The results of the preliminary study that are presented here, which focused on interactions between the insect gut microbiota and mutualistic bacteria of entomopathogenic nematodes, show that bacteria inhabiting the gut of insects might play a key role in insect resistance to entomopathogenic nematode pressure.
- ItemComparative nanopore sequencing-based evaluation of the midgut microbiota of the summer chafer (Amphimallon solstitiale L.) associated with possible resistance to entomopathogenic nematodes(MDPI, 2022-03-15) Sajnaga, Ewa; Skowronek, Marcin; Kalwasińska, Agnieszka; Kazimierczak, Waldemar; Lis, Magdalena; Jach, Monika Elżbieta; Wiater, AdrianRoot-feeding Amphimallon solstitiale larvae and certain other scarab beetles are the main soil-dwelling pests found in Europe, while entomopathogenic nematodes (EPN) have been used as a biocontrol agent against these species. Our study provides the first detailed characterization of the bacterial community of the midgut in wild A. solstitiale larvae, based on the nanopore sequencing of the 16S rRNA gene. In the whole dataset, we detected 2586 different genera and 11,641 species, with only 83 diverse bacterial genera shared by all studied individuals, which may represent members of the core midgut microbiota of A. solstitiale larvae. Subsequently, we compared the midgut microbiota of EPN-resistant and T0 (prior to EPN exposure) individuals, hypothesizing that resistance to this parasitic infection may be linked to the altered gut community. Compared to the control, the resistant insect microbiota demonstrated lower Shannon and Evenness indices and significant differences in the community structure. Our studies confirmed that the gut microbiota alternation is associated with resistant insects; however, there are many processes involved that can affect the bacterial community. Further research on the role of gut microbiota in insect-parasitic nematode interaction may ultimately lead to the improvement of biological control strategies in insect pest management.
- ItemEvolution and taxonomy of nematode-associated entomopathogenic bacteria of the genera Xenorhabdus and Photorhabdus: an overview(Springer Nature, 2020-01-10) Sajnaga, Ewa; Kazimierczak, WaldemarEntomopathogenic bacteria from the genera Photorhabdus and Xenorhabdus are closely related Gram-negative bacilli from the family Enterobacteriaceae (γ-Proteobacteria). They establish obligate mutualistic associations with soil nematodes from the genera Steinernema and Heterorhabditis to facilitate insect pathogenesis. The research of these two bacterial genera is focused mainly on their unique interactions with two different animal hosts, i.e. nematodes and insects. So far, studies of the mutualistic bacteria of nematodes collected from around the world have contributed to an increase in the number of the described Xenorhabdus and Photorhabdus species. Recently, the classification system of entomopatogenic nematode microsymbionts has undergone profound revision and now 26 species of the genus Xenorhabdus and 19 species of the genus Photorhabdus have been identified. Despite their similar life style and close phylogenetic origin, Photorhabdus and Xenorhabdus bacterial species differ significantly in e.g. the nematode host range, symbiotic strategies for parasite success, and arrays of released antibiotics and insecticidal toxins. As the knowledge of the diversity of entomopathogenic nematode microsymbionts helps to enable the use thereof, assessment of the phylogenetic relationships of these astounding bacterial genera is now a major challenge for researchers. The present article summarizes the main information on the taxonomy and evolutionary history of Xenorhabdus and Photorhabdus, entomopathogenic nematode symbionts.
- ItemGum arabic/gelatin and water-soluble soy polysaccharide/gelatin blend films as carriers of astaxanthin - a comparative study of the kinetics of release and antioxidant properties(MDPI, 2021-03-28) Kowalczyk, Dariusz; Łupina, Katarzyna; Kazimierczak, WaldemarPolymer blending and incorporation of active substances offer a possibility of generation of novel packaging materials with interesting features. Astaxanthin is one of the most powerful antioxidants. Hence, in this study, water-soluble AstaSana astaxanthin (AST) was incorporated into 75/25 gum arabic/gelatin (GAR75/GEL25) and water-soluble soy polysaccharides/gelatin (WSSP75/GEL25) blend films in different concentrations (0, 0.25%, 0.5%, 1%). Microscope images showed good compatibility between the polysaccharides and GEL. Basing on time required for 50% release, the WSSP-based film exhibited an approximately four-fold slower release rate (t50% = 65.16–142.80 min) than the GAR-based film (t50% = 14.64–34.02 min). This result was mainly ascribed to the slower dissolution of the WSSP-based carrier. The faster release rate of the GAR-based films resulted in stronger antioxidant activity (quarter-scavenging time (t25%ABTS) = 0.22–7.51 min) in comparison to the WSSP-based films (t25%ABTS = 0.91–12.94 min). The increase in the AST concentration was accompanied by gradually reduced solubility and the release rate. It is possible that the increasing number of starch granules (from the AST formulation) acted as a dissolution blocking agent. In general, the WSSP75/GEL25 film displayed the most linear (the Zero-order similar) release profile. So, this carrier has potential for release of AST at a quasi-constant speed.
- ItemIs soil management system really important? comparison of microbial community diversity and structure in soils managed under organic and conventional regimes with some view on soil properties(PLoS, 2021-09-09) Rachwał, Kamila; Gustaw, Klaudia; Kazimierczak, Waldemar; Waśko, AdamThe fertility and productive value of soil are closely related to the physical and chemical properties of the soil as well as its biological activity. This activity is related to the intensity of microbially catalysed processes of transformation of organic and mineral substances contained in the soil. These processes are closely correlated with the abundance and biodiversity of soil microorganisms, especially bacteria, and the activity of enzymes produced by them. In this article we have compared some physicochemical properties of soil derived from conventional and organic farms and microbial communities inhabiting these ecosystems. We aim to investigate whether the soil management regime affects the abundance and diversity of these environments in terms of bacteria. Some differences in microbial communities were observed, but the rhizosphere of plants from organic and conventional soils does not harbour separate microbiomes. Albeit, the method of fertilization influences the diversity of soil microorganisms. A greater diversity of bacteria was observed in soils from farms where organic fertilizers were applied. Soil pH and activity of some soil enzymes were also shown to differ between organic and conventional soil cropping systems.
- ItemNanopore-sequencing characterization of the gut microbiota of Melolontha melolontha larvae: contribution to protection against entomopathogenic nematodes?(MDPI, 2021-03-25) Sajnaga, Ewa; Skowronek, Marcin; Kazimierczak, Waldemar; Ferenc, Karolina; Lis, Magdalena; Wiater, AdrianThis study focused on the potential relationships between midgut microbiota of the common cockchafer Melolontha melolontha larvae and their resistance to entomopathogenic nematodes (EPN) infection. We investigated the bacterial community associated with control and unsusceptible EPN-exposed insects through nanopore sequencing of the 16S rRNA gene. Firmicutes, Proteobacteria, Actinobacteria, and Bacteroidetes were the most abundant bacterial phyla within the complex and variable midgut microbiota of the wild M. melolontha larvae. The core microbiota was found to include 82 genera, which accounted for 3.4% of the total number of identified genera. The EPN-resistant larvae differed significantly from the control ones in the abundance of many genera belonging to the Actinomycetales, Rhizobiales, and Clostridiales orders. Additionally, the analysis of the microbiome networks revealed different sets of keystone midgut bacterial genera between these two groups of insects, indicating differences in the mutual interactions between bacteria. Finally, we detected Xenorhabdus and Photorhabdus as gut residents and various bacterial species exhibiting antagonistic activity against these entomopathogens. This study paves the way to further research aimed at unravelling the role of the host gut microbiota on the output of EPN infection, which may contribute to enhancement of the efficiency of nematodes used in eco-friendly pest management.
- ItemScreening and molecular identification of bacteria from the midgut of Amphimallon solstitiale larvae exhibiting antagonistic activity against bacterial symbionts of entomopathogenic nematodes(MDPI, 2021-11-05) Skowronek, Marcin; Sajnaga, Ewa; Kazimierczak, Waldemar; Lis, Magdalena; Wiater, AdrianEntomopathogenic nematodes (Rhabditida: Steinernematidae and Heterorhabditidae) are a group of organisms capable of infecting larvae of insects living in soil, including representatives of the family Scarabaeidae. Their insecticidal activity is related to the presence of symbiotic bacteria Xenorhabdus spp. or Photorhabdus spp. in the alimentary tract, which are released into the insect body, leading to its death caused by bacterial toxins and septicemia. Although the antibacterial activities of symbionts of entomopathogenic nematodes have been well described, there is insufficient knowledge of the interactions between these bacteria and microorganisms that naturally inhabit the alimentary tract of insects infested by nematodes. In this study, 900 bacterial strains isolated from midgut samples of Amphimallon solstitiale larvae were tested for their antagonistic activity against the selected five Xenorhabdus and Photorhabdus species. Cross-streak tests showed significant antibacterial activity of 20 isolates. These bacteria were identified as Bacillus [Brevibacterium] frigoritolerans, Bacillus toyonensis, Bacillus wiedmannii, Chryseobacterium lathyri, Chryseobacterium sp., Citrobacter murliniae, Enterococcus malodoratus, Paenibacillus sp., Serratia marcescens and Serratia sp. Since some representatives of the intestinal microbiota of A. solstitiale are able to inhibit the growth of Xenorhabdus and Photorhrhabdus bacteria in vitro, it can be assumed that this type of bacterial interaction may occur at certain stages of insect infection by Steinernema or Heterorhabditis nematodes.
- ItemSteinernema sandneri n. sp. (Rhabditida: Steinernematidae), a new entomopathogenic nematode from Poland(Exeley, 2021-05-21) Lis, Magdalena; Sajnaga, Ewa; Skowronek, Marcin; Wiater, Adrian; Rachwał, Kamila; Kazimierczak, WaldemarA new species of entomopathogenic nematodes, Steinernema sandneri n. sp., was recovered by baiting from Poland. Its morphological traits indicate that the new species is a member of the feltiae-kraussei group. A body length of 843 (708-965) μm., a more anterior position of excretory pore (56 μm), and the lower D% value (40 vs. >46) discriminate this species from most of the other group members. The first-generation males of S. sandneri n. sp. can be distinguished from the other clade members by a 60 μm long spicule, a relatively long gubernaculum (GS%=79), and the position of the excretory pore (80 μm). Phylogenetic analysis of the ITS rDNA, D2D3 of 28S rDNA, and cox1 sequences confirmed that S. sandneri n. sp. is a new species of the feltiae-kraussei group, closely related to S. kraussei and S. silvaticum.