Survival Rate Of Nematodes Research Articles

An automatic recognition method of nematode survival rate based on bright field and dark field experimental images.

The survival rate of experimental animals is a very important index in chemical toxicity evaluation experiments. The calculation of nematode survival rate is used in many experiments. Traditional survival rate quantification methods require manual counting. This is a time-consuming and laborious work when using 384-well plate for high-throughput chemical toxicity assessment experiments. At present, there is a great need for an automatic method to identify the survival rate of nematodes in the experiment of chemical toxicity evaluation. We designed an automatic nematode survival rate recognition method by combining the bright field experimental image of nematodes and the dark field image of nematodes which is captured after adding Propidium Iodide dye, and used it to calculate the nematode survival rate in different chemical environments. Experiment results show that the survival rate obtained by our automatic counting method is very similar to the survival rate obtained by manual counting. Through several different chemical experiments, we can see that chemicals with different toxicity have different effects on the survival rate of nematodes. And the survival rate of nematodes under different chemical concentrations has an obvious gradient trend from high concentration to low concentration. In addition, our method can quantify the motility of nematodes. There are also significant differences in the motility of nematodes cultured in different chemical environments. Moreover, the nematode motility under different chemical concentrations showed an obvious gradient change trend from high concentration to low concentration. Our study provides an accurate and efficient nematode survival rate recognition method for chemical toxicology research.

Functional analysis of 3 genes in xenobiotic detoxification pathway of Bursaphelenchus xylophilus against matrine

Bursaphelenchus xylophilus is the causative agent of pine wilt disease. It has caused devastating damage to ecosystems worldwide, owing to the characteristic of being widely spread and uncontrollable. However, the current methods of control are mainly based on pesticides, which can cause irreversible damage to the ecosystem. Therefore, the search for new drug targets and the development of environmentally friendly nematicides is especially valuable. In this study, three key genes of the xenobiotic detoxification pathways were cloned from B. xylophilus, which were subsequently subjected to bioinformatic analysis. The bioassay experiment was carried out to determine the concentration of matrine required for further tests. Subsequently, enzyme activity detection and three gene expression pattern analysis were performed on matrine treated nematodes. Finally, RNA interference was conducted to verify the functions carried out by the three genes in combating matrine. The results indicated that cytochrome P450 and glutathione S-transferase of B. xylophilus were activated by matrine, which induced high expression of BxCYP33C4, BxGST1, and BxGST3. After RNA interference of three genes of B. xylophilus, the sensitivity of B. xylophilus to matrine was increased and the survival rate of nematodes was reduced to various degrees in comparison to the control group. Overall, the results fully demonstrated that BxCYP33C4, BxGST1, and BxGST3 are valuable drug targets for B. xylophilus. Furthermore, the results suggested that matrine has value for development and exploitation in the prevention and treatment of B. xylophilus.

Ginseng volatile oil prolongs the lifespan and healthspan of Caenorhabditis elegans.

Ginseng volatile oil (GVO) is one of the main components of ginseng and has antibacterial and anti-inflammatory properties. In this study, gas chromatography–mass spectrometry (GC–MS) was applied to characterize GVO chemical composition, and 73 volatile components were detected from GVO. Caenorhabditis elegans was used as animal model to further elucidate the antioxidant and anti-aging effects of GVO in vivo. The results suggested that GVO significantly prolonged the lifespan of C. elegans and promoted its health without damaging its reproductive capacity. In addition, GVO increased the antioxidant capacity and survival rate of nematodes after heat shock. Transcriptional sequencing showed that autophagy-related genes atg-4.2, atg-7, lgg-2, and cyd-1 were up-regulated, and superoxide dismutase 1 (sod-1) expression was increased after GVO pretreatment. Considering the role of autophagy and antioxidant in aging, the expression of autophagy substrate P62 protein in BC12921 strain was analyzed and found to decrease by more than 50.00% after treatment with GVO. In addition, the lifespan of SOD-1 mutant nematodes was not significantly different from that of the control group. SOD activity and autophagy were activated, which is a clear expression of hormesis. All these results suggest that GVO prolongs the lifespan and healthspan of C. elegans, and its biological functions may be related to hormesis.Supplementary InformationThe online version contains supplementary material available at 10.1007/s10522-022-09956-z.

COMPATIBILITY OF NIMBECIDINE AND TRICEL WITH ENTOMOPATHOGENIC NEMATODE, HETERORHABDITIS BACTERIOPHORA

A laboratory study was conducted to study of compatibility of entomopathogenic nematode, Heterorhabditis bacteriophora with insecticides currently used against cutworm (Agrotis ipsilon). To evaluate the nematode survival rate, infective juveniles (IJs) H. bacteriophora were exposed to Nimbecidine and Tricel at three different concentrations for 3 h,6 h, 12 h and 24 h. The surviving nematodes were used to determine the infectivity, penetration rate and reproduction potential in last instars cutworm larvae. Data showed that the survival rate of nematodes exposed to Nimbecidine and Tricel was 53% and 50% respectively. H. bacteriophora was found to be virulent with the A. ipsilon mortality (60 % to 70%).Penetration rate of IJs of H.bacteriophora in A. ipsilon larvae reached 55.5% when exposed to Nimbecidine at 0.125%. H.bacteriophora was more susceptible to chlorpyriphos than Nimbecidine. H. bacteriophora infectious ability and reproduction rate in A. ipsilon was found to be reduced in exposure of chlorpyriphos

Small interfering RNAs targeting agrA and sarA attenuate pathogenesis of Staphylococcus aureus in Caenorhabditis elegans.

The use of small interfering RNA (siRNA) gene silencing is a promising therapeutic option as it does not impose selective pressure on bacteria that is often associated with the development of resistance. The study assessed the effect of siRNA targeted to sarA and agrA in S. aureus and the relationship between the transcriptional response, biofilm formation and pathogenicity. siRNAs designed against agrA and sarA were electroporated into methicillin-resistant and methicillin-susceptible S. aureus strains. mRNA levels, growth kinetics, biofilm formation and minimal inhibitory concentration were measured. Efficacy of siRNA in bacteria was assessed using survival assays in a C. elegans model. Differences in gene expression before and after siRNA treatment were anaysed using the paired t-test, while the log rank test was used to assess the significance of any difference among survival rates of nematodes. Biofilm formation decreased significantly in siRNA treated strains and growth rates of siRNA treated strains were significantly higher compared to untreated strains. We observed significant decreases in the transcriptional response in siRNA treated strains, with concomitant significant increases in the lifespan of C. elegans worms exposed to siRNA-treated versus untreated strains. siRNA targeted to agrA and sarA lowered mRNA transcription and pathogenicity of S. aureus.

Application of Entomopathogenic Nematodes/Metal Organic Frameworks/Sodium Alginate Composite Microspheres in the Control of Termites

In order to provide scientific basis for control of termites, the control of termites using entomopathogenic nematodes/Metal organic Frameworks/sodium alginate composite microspheres was herein studied. Firstly, four MOFs with antibacterial properties, MOF-199, ZIF-67, MOF-5 and ZIF-8, were synthesized, and then entomopathogenic nematodes/MOFs/sodium alginate composite microspheres were prepared. Under laboratory culture conditions, the survival rate of nematodes in the particles and mortality rate against termites were determined. Entomopathogenic nematodes/MOFs/sodium alginate composite microspheres were found to have good antibacterial properties, which can prolong the action time for the particles. MOFs had no adverse effect on survival rate of nematodes, and the lethal effect of MOFs on termites reached up to 85%. We can safely draw the conclusion that the entomopathogenic nematodes/MOFs/sodium alginate composite microspheres can effectively trap and kill termites. This study will provide a new idea for development of biological insecticides and control of termites.

Capsule-C: an improved Steinernema carpocapsae capsule formulation for controlling Agrotis ipsilon Hufnagel (Lepidoptera: Noctuidae)

BackgroundEntomopathogenic nematodes (EPNs) have long been used for controlling soil-dwelling insects. Steinernema carpocapsae HB310, previously showed a high virulence against many pests including Agrotis ipsilon Hufnagel (Lepidoptera: Noctuidae). Due to the lack of durable formulations, up until now, S. carpocapsae HB310 has thus far been prevented from use in large-scale farming. The present study aimed to get a better EPNs capsule formulation suitable for long-term storage and effective application.ResultsAn improved EPNs capsule formulation, herein named: Capsule-C was prepared by the following composition: Solution I: 18% glycerol, 0.075% formaldehyde, 1% sodium alginate, 0.2% xanthan gum, 0.5% potassium sorbate, 9% glucose, 2% fructose, 2% sucrose, and the remainder was distilled water. The nematodes suspension was added to the alginate mixture in 2 × 104 IJs/mL; Solution II: 18% glycerol, 0.075% formaldehyde, 0.5% calcium chloride, 0.5% potassium sorbate, with the remainder being distilled water. After storage for 180 days at 16 °C and 100% RH, the survival rate of nematodes in Capsule-C was 75.68 ± 0.48% and the nematodes caused 82.33 ± 1.45% mortality in the 5th instar larvae of Galleria mellonella. A. ipsilon larvae preferred to chew and ingest Capsule-C due to the addition of the glucose compound. The feeding rate of A. ipsilon larvae on Capsule-C reached to 100% within 24 h and the larval mortality of A. ipsilon was 90.48 ± 6.35%.ConclusionEPNs-containing capsules were as effective as sprayed EPNs in water solution at killing A. ipsilon. These results will provide ideas to acquire a stable and efficient EPNs capsule formulation and further promote the application of environmental friendly biological pesticides.

Manufacturing of Granular Matrix with Diatomacious earth, Lime and Sugarcane Bagasse ash to Encapsulate Entomopathogenic Nematodes (Steinernema glaseri)

An alternative for pest control in corn production is the use of biopesticides formulated with entomopathogenic nematodes encapsulated in granular matrices. However, it has been observed that the granular matrices´ lack of resistance to handling and transport causes their deterioration, therefore limiting their use. This study analyzes the use of lime and sugarcane bagasse ash as stabilizer additives in granular matrices made of diatomite earth in order to improve the survival rate of Entomopathogenic Nematodes. Six formulations were prepared and evaluated at 3, 12, 21 and 30 days of age. A total of 192 granular matrices were produced, 144 for axial compressive strength testing and 48 for the estimation of the survival rate of the nematodes. In order to carry out the last test, Steinernema glaseri nematodes were encapsulated. The results indicate that the granular matrices made with the combination of 88% diatomaceous earth plus 6% lime and 6% sugarcane bagasse ash have adequate hardness (mechanical strength) for handling and transportation purposes; in addition, at 30 days of age the survival rate was the highest obtained when compared to the granular matrices of all tested formulations

Impact of insecticides on the efficacy of entomopathogenic nematodes against the diamondback moth Plutella xylostella (L.) (Lepidoptera: Plutellidae) on cabbage in northern Benin

Description of the subject. The article deals with the study of compatibility of entomopathogenic nematode (EPN) species with insecticides currently used against diamondback moth (DBM) Plutella xylostella (L.) in northern Benin Objectives. The aim of this work was to determine the impact of five insecticides on the efficacy of entomopathogenic nematodes against DBM larvae. Method. Infective juveniles (IJs) of two EPN species (Steinernema sp. 83a and Heterorhabditis sonorensis KF723827) were exposed to five insecticides used against P. xylostella larvae in Djougou for 48 h. The number of surviving nematodes was used to infest DBM larvae. The experiment was carried out under laboratory and semi-field conditions. Results. The bioassays carried out in laboratory showed that the survival rate of nematodes exposed to KARATE 2.5 WG, LAMBDA SUPER 2.5EC or neem oil ranges between 95% and 98%. In the treated plots, Steinernema sp. 83a was the most virulent with the highest P. xylostella mortality (87% at 50 IJs·cm-2 after 24 h) while only 35% larval mortality was recorded for H. sonorensis applied at the same dose. Population density of nematodes which penetrated DBM larvae reached 9 ± 3I IJs·larva-1 for H. sonorensis KF723827 and 6 ± 2 IJs·larva-1 for Steinernema sp. 83a. In cadaver of DBM, nematode reproduction did not appear to be affected by the contact with insecticides. Conclusions. Based on our research, we conclude that the three insecticides did not affect EPNs efficiency and could be used in combination against DBM.

Brevinin-2 Drug Family—New Applied Peptide Candidates Against Methicillin-Resistant Staphylococcus aureus and Their Effects on Lys-7 Expression of Innate Immune Pathway DAF-2/DAF-16 in Caenorhabditis elegans

The issue of Staphylococcus aureus (MRSA) developing a resistance to drugs such as methicillin has long been the focus for new drug development. In recent years, antimicrobial peptides, such as small molecular peptides with broad-spectrum antibacterial activity and special antibacterial mechanism, have shown a strong medicinal potential. In particular, the Brevinin-2 family has been shown to have a significant inhibitory effect against gram-positive bacteria (G+). In this study, we researched the influence of MRSA on the behavior and survival rate of nematodes. We established an assay of Caenorhabditis elegans–MRSA antimicrobial peptides to screen for new potent anti-infective peptides against MRSA. From the Brevinin-2 family, 13 peptides that had shown strong effects on G+ were screened for their ability to prolong the lifespan of infected worms. Real-time Polymerase Chain Reaction (PCR) tests were used to evaluate the effect on the innate immune pathway dauer formation defective (DAF)-2/DAF-16 of C. elegans. The assay successfully screened and filtered out four of the 13 peptides that significantly improved the survival rate of MRSA-infected worms. The result of real-time PCR indicated that the mRNA and protein expression levels of lys-7 were consistently upregulated by being treated with four of the Brevinin-2 family. The Brevinin-2 family peptides, including Brevinin-2, Brevinin-2-OA3, Brevinin-2ISb, and Brevinin-2TSa, also played an active role in the DAF-2/DAF-16 pathway in C. elegans. We successfully demonstrated the utility of anti-infective peptides that prolong the survival rate of the MRSA-infected host and discovered the relationship between antibacterial peptides and the innate immune system of C. elegans. We demonstrated the antimicrobial effects of Brevinin-2 family peptides, indicating their potential for use as new drug candidates against MRSA infections.

Treatment of Nematode in Soil Using Surface Barrier Discharge Ozone Generator

One of the practical usages of plasma in large-scaled agriculture is the indirect application of plasma by irradiating plasma derivative radicals. Since agricultural chemicals cause environmental problems, such as the generation of resistant bacteria and the increase of toxic residuals, we have proposed the extermination of soil worms and the acceleration of plant’s growth by the ozone treatment of soil. Here, we propose soil treatment using ozone, generated by a surface barrier discharge, as both roles of pesticides and fertilizers. To investigate the effectiveness of gaseous ozone for the extermination of soil worms, this paper researched the influence of ozone on representative nematodes of C. elegans . Furthermore, to evaluate the practical effectiveness of the ozone treatment of natural nematodes living in agricultural soil, natural andosol was treated by ozone and the survival rate of nematodes was analyzed statistically using the Baermann funnel extraction.

Fermented Brown Sugar Residue Prolongs the <i>Caenorhabditis elegans</i> Lifespan <i>via</i> DAF-16

Purification of biomass ethanol from the products of brown sugar yeast-fermentation produces a large amount of residue. This fermentation residue contains abundant brown sugar-derived nutrients and is mainly used as compost or livestock feed. However, the in vivo physiological effects of oral residue ingestion are not known. The purpose of this study was to elucidate the physiological action and molecular mechanism of fermented brown sugar residue in nematode stress tolerance, aging, and lifespan using Caenorhabditis elegans. Fermented brown sugar residue was divided into two layers, supernatant and precipitate, and each was given to nematodes. Analysis of motility and survival rate under thermal stress revealed reduced mobility and increased survival rate following treatment with fermented brown sugar residue. The survival rate of nematodes under 1% H2O2 was markedly increased by the residue and mitochondrial membrane depolarization was induced and mitochondrial radical oxygen species levels increased. Furthermore, aging dependent reduction of motility was suppressed, and the average life span of nematodes was extended by treatment with fermented brown sugar residue. Moreover, the effects of fermented brown sugar residue on stress tolerance, lifespan elongation, and decreased aging dependent momentum reduction were lost in the daf-16 mutant. Taken together, our results show that the various physiological actions of fermented brown sugar residue, including stress tolerance and lifespan extension, occur via DAF-16.

Anti-aging properties of Ribes fasciculatum in Caenorhabditis elegans

The present study investigated the effects and underlying mechanism of ethylacetate fraction of Ribes fasciculatum (ERF) on the lifespan and stress tolerance using a Caenorhabditis elegans model. The longevity activity of ERF was determined by lifespan assay under normal culture condition. The survival rate of nematodes under various stress conditions was assessed to validate the effects of ERF on the stress tolerance. To determine the antioxidant potential of ERF, the superoxide dismutase (SOD) activities and intracellular reactive oxygen species (ROS) levels were investigated. The ERF-mediated change in SOD-3 expression was examined using GFP-expressing transgenic strain. The effects of ERF on the aging-related factors were investigated by reproduction assay and pharyngeal pumping assay. The intestinal lipofuscin levels of aged nematodes were also measured. The mechanistic studies were performed using selected mutant strains. Our results indicated that ERF showed potent lifespan extension effects on the wild-type nematode under both normal and various stress conditions. The ERF treatment also enhanced the activity and expression of superoxide dismutase (SOD) and attenuated the intracellular ROS levels. Moreover, ERF-fed nematodes showed decreased lipofuscin accumulation, indicating ERF might affect age-associated changes in C. elegans. The results of mechanistic studies indicated that there was no significant lifespan extension in ERF-treated daf-2, age-1, sir-2.1, and daf-16 null mutants, suggesting that they were involved in ERF-mediated lifespan regulation. In conclusion, R. fasciculatum confers increased longevity and stress resistance in C. elegans via SIR-2.1-mediated DAF-16 activation, dependent on the insulin/IGF signaling pathway.

Virulence variations in Shigella and enteroinvasive Escherichia coli using the Caenorhabditis elegans model.

Shigella species and enteroinvasive Escherichia coli (EIEC) belong to the same species genetically, with remarkable phenotypic and genomic similarities. Shigella is the main cause of bacillary dysentery with around 160 million annual cases, while EIEC generally induces a milder disease compared to Shigella. This study aimed to determine virulence variations between Shigella and EIEC using the nematode Caenorhabditis elegans as a model host. Caenorhabditis elegans killing- and bacterial colonization assays were performed to examine the potential difference in virulence between Shigella and EIEC strains. Statistically significant difference in the survival rates of nematodes was demonstrated, with Shigella causing death at 88.24 ± 1.20% and EIEC at 94.37 ± 0.70%. The intestinal load of bacteria in the nematodes was found to be 7.65 × 10(4) ± 8.83 × 10(3) and 2.92 × 10(4) ± 6.26 × 10(3) CFUml(-1) per nematode for Shigella and EIEC, respectively. Shigella dysenteriae serotype 1 which carries the Shiga toxin showed the lowest nematode survival rate at 82.6 ± 3.97% and highest bacterial colonization of 1.75 × 10(5) ± 8.17 × 10(4) CFUml(-1), whereas a virulence plasmid-negative Shigella strain demonstrated 100 ± 0% nematode survival and lowest bacterial accumulation of 1.02 × 10(4) ± 7.23 × 10(2) CFUml(-1). This study demonstrates C. elegans as an effective model for examining and comparing Shigella and EIEC virulence variation.