A Facile and Modified Scheme for Synchronization and Isolation of Nematode Eggs

Agriculture ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 676
Author(s):  
Satish Kumar Rajasekharan ◽  
Chaitany Jayprakash Raorane ◽  
Jintae Lee
Keyword(s):  
Caenorhabditis Elegans ◽  
Bursaphelenchus Xylophilus ◽  
In Vitro Studies ◽  
Parasitic Nematodes ◽  
Distilled Water ◽  
Agricultural Crop ◽  
Laboratory Setting ◽  
Straightforward Method ◽  
Nematode Eggs

Nematodes are common pests that damage agricultural crop plants. Some of them are beneficial while others are parasitic and harmful to plants, animals and humans. Several in vitro studies have aimed to develop chemicals to kill parasitic nematodes, while others have been conducted to use beneficial nematodes as biocontrol agents. However, the preparation of large quantities of nematode eggs in a laboratory setting is challenging. Traditional egg isolation protocols involve the use of sieves to filter eggs or the use of chemicals that can be harmful to nematodes while isolating the eggs. Our method utilizes 1.5 × 1.5 cm sized chunks of bacterial or fungal feed to lure nematodes. A subsequent gentle washing of the consumed chunk with distilled water provides a rapid and straightforward method of collecting eggs in 6-well polystyrene plates and removing unwanted nematodes. Approximately 4000 Bursaphelenchus xylophilus eggs from a fungal chunk and 2400 Caenorhabditis elegans eggs from a bacterial chunk were obtained when tested. This study shows a protocol for the isolation of eggs and synchronization of nematode stages that is relatively straightforward, rapid, eco-friendly, and efficient. The protocol also provides a chemical-free and a reliable, simple means of separating eggs from adults and induces the synchronization of nematodes based on the simple concept that gravid nematodes can be provoked to lay eggs by providing additional feed.

scholarly journals Natural variation in Caenorhabditis elegans responses to the anthelmintic emodepside

2021 ◽  
Author(s):  
Janneke Wit ◽  
Steffen R. Hahnel ◽  
Briana C. Rodriguez ◽  
Erik Andersen
Keyword(s):  
Caenorhabditis Elegans ◽  
Mode Of Action ◽  
Natural Variation ◽  
Treatment Strategies ◽  
Parasitic Nematodes ◽  
Anthelmintic Drug ◽  
C Elegans ◽  
Hormetic Effect ◽  
Filarial Nematodes

Treatment of parasitic nematode infections depends primarily on the use of anthelmintics. However, this drug arsenal is limited, and resistance against most anthelmintics is widespread. Emodepside is a new anthelmintic drug effective against gastrointestinal and filarial nematodes. Nematodes that are resistant to other anthelmintic drug classes are susceptible to emodepside, indicating that the emodepside mode of action is distinct from previous anthelmintics. The laboratory-adapted Caenorhabditis elegans strain N2 is sensitive to emodepside, and genetic selection and in vitro experiments implicated slo-1, a BK potassium channel gene, in emodepside mode of action. In an effort to understand how natural populations will respond to emodepside, we measured brood sizes and developmental rates of wild C. elegans strains after exposure to the drug and found natural variation across the species. Some variation in emodepside responses can be explained by natural differences in slo-1. This result suggests that other genes in addition to slo-1 underlie emodepside resistance in wild C. elegans strains. Additionally, all assayed strains have higher offspring production in low concentrations of emodepside (a hormetic effect), which could impact treatment strategies. We find that natural variation affects emodepside sensitivity, supporting the suitability of C. elegans as a model system to study emodepside responses across parasitic nematodes.

scholarly journals In Vitro Efficacy of Aqueous and Methanol Extract of Cassia siamea Against the Motility of Caenorhabditis elegans

2020 ◽  
Vol 31 (3) ◽  
pp. 145-159
Author(s):  
Haladu Ali Gagman ◽  
Nik Ahmad Irwan Izzauddin Nik Him ◽  
Hamdan Ahmad ◽  
Shaida Fariza Sulaiman ◽  
Rahmad Zakaria ◽  
...  
Keyword(s):  
Caenorhabditis Elegans ◽  
Methanol Extract ◽  
Anthelmintic Resistance ◽  
Gastrointestinal Nematode ◽  
Parasitic Nematodes ◽  
Cassia Siamea ◽  
Methanol Extracts ◽  
C Elegans ◽  
Significant Difference

Gastrointestinal nematode infections can cause great losses in revenue due to decrease livestock production and animal death. The use of anthelmintic to control gastrointestinal nematode put a selection pressure on nematode populations which led to emergence of anthelmintic resistance. Because of that, this study was carried out to investigate the efficacy of aqueous and methanol extract of Cassia siamea against the motility of C. elegans Bristol N2 and C. elegans DA1316. Caenorhabditis elegans Bristol N2 is a susceptible strain and C. elegans DA1316 is an ivermectin resistant strain. In vitro bioassay of various concentrations of (0.2, 0.6, 0.8, 1.0 and 2.0 mg mL–1) aqueous and methanol extracts of C. siamea was conducted against the motility of L4 larvae of C. elegans Bristol N2 and C. elegans DA1316. The L4 larvae were treated with 0.02 μg mL–1 of ivermectin served as positive control while those in M9 solution served as negative control. The activity of the extracts was observed after 24 h and 48 h. A significant difference was recorded in the extract performance compared to control at (P < 0.001) after 48 h against the motility of the larvae of both strains. The methanol extracts inhibited the motility of C. elegans Bristol N2 by 86.7% as well as DA1316 up to 84.9% at 2.0 mg mL–1 after 48 h. The methanol extract was more efficient than aqueous extract (P < 0.05) against the motility of both strains of C. elegans. Cassia siamea may be used as a natural source of lead compounds for the development of alternative anthelmintic against parasitic nematodes as well ivermectin resistant strains of nematodes.

scholarly journals Morphologically distinct microtubule ends in the mitotic centrosome of Caenorhabditis elegans

2003 ◽  
Vol 163 (3) ◽  
pp. 451-456 ◽  
Author(s):  
Eileen T. O'Toole ◽  
Kent L. McDonald ◽  
Jana Mäntler ◽  
J. Richard McIntosh ◽  
Anthony A. Hyman ◽  
...  
Keyword(s):  
Caenorhabditis Elegans ◽  
Dynamic Behavior ◽  
High Voltage ◽  
Electron Tomography ◽  
In Vitro Studies ◽  
Embryonic Cells ◽  
Nematode Caenorhabditis Elegans ◽  
Voltage Electron ◽  
High Voltage Electron

During mitosis, the connections of microtubules (MTs) to centrosomes and kinetochores are dynamic. From in vitro studies, it is known that the dynamic behavior of MTs is related to the structure of their ends, but we know little about the structure of MT ends in spindles. Here, we use high-voltage electron tomography to study the centrosome- and kinetochore-associated ends of spindle MTs in embryonic cells of the nematode, Caenorhabditis elegans. Centrosome-associated MT ends are either closed or open. Closed MT ends are more numerous and are uniformly distributed around the centrosome, but open ends are found preferentially on kinetochore-attached MTs. These results have structural implications for models of MT interactions with centrosomes.

scholarly journals Mortality and movement behaviour of Bursaphelenchus xylophilus under different dosages of copper sulphate

2013 ◽  
Vol 49 (No. 2) ◽  
pp. 98-103 ◽  
Author(s):  
Q.Q. Tan ◽  
H.Y. Wu ◽  
S.X. Jiang ◽  
H. Bing
Keyword(s):  
Locomotor Activity ◽  
Copper Sulphate ◽  
Bursaphelenchus Xylophilus ◽  
Distilled Water ◽  
Movement Behaviour ◽  
Median Lethal Concentration ◽  
Locomotor Behaviour ◽  
In Vitro Exposure ◽  
Harmful Effects

The effects of in vitro exposure to different concentrations of copper sulphate for different durations on the mortality and locomotor behaviour of Bursaphelenchus xylophilus were evaluated. Copper sulphate showed strong effects against the nematode B. xylophilus at a low concentration (5 mg/l). The median lethal concentration (LC<sub>50</sub>) at 8, 24, 48, and 72 h were 31.634, 7.353, 6.557, and 5.568 mg/l, respectively. Mortality rate of B. xylophilus after treatment with various concentrations of copper sulphate for 6, 8, 24, 48, and 72 h were significantly different compared with control animals exposed to distilled water (P &lt; 0.01). Locomotor activity of the nematodes decreased with increased duration of exposure and was completely lost after 8 h of exposure to 100 mg/l copper sulphate. Our results indicate that copper sulphate is effective against B. xylophilus and restricts their harmful effects on plants at least in part by inhibiting their movement.&nbsp; &nbsp;

scholarly journals Practical High-Throughput Method to Screen Compounds for Anthelmintic Activity against Caenorhabditis elegans

Molecules ◽  
2021 ◽  
Vol 26 (14) ◽  
pp. 4156
Author(s):  
Aya C. Taki ◽  
Joseph J. Byrne ◽  
Peter R. Boag ◽  
Abdul Jabbar ◽  
Robin B. Gasser
Keyword(s):  
Caenorhabditis Elegans ◽  
Small Molecules ◽  
High Throughput ◽  
High Throughput Screening ◽  
Anthelmintic Activity ◽  
Infrared Light ◽  
Parasitic Nematodes ◽  
Screening Assay ◽  
High Throughput Screening Assay

In the present study, we established a practical and cost-effective high throughput screening assay, which relies on the measurement of the motility of Caenorhabditis elegans by infrared light-interference. Using this assay, we screened 14,400 small molecules from the “HitFinder” library (Maybridge), achieving a hit rate of 0.3%. We identified small molecules that reproducibly inhibited the motility of C. elegans (young adults) and assessed dose relationships for a subset of compounds. Future work will critically evaluate the potential of some of these hits as candidates for subsequent optimisation or repurposing as nematocides or nematostats. This high throughput screening assay has the advantage over many previous assays in that it is cost- and time-effective to carry out and achieves a markedly higher throughput (~10,000 compounds per week); therefore, it is suited to the screening of libraries of tens to hundreds of thousands of compounds for subsequent evaluation and development. The present phenotypic whole-worm assay should be readily adaptable to a range of socioeconomically important parasitic nematodes of humans and animals, depending on their dimensions and motility characteristics in vitro, for the discovery of new anthelmintic candidates. This focus is particularly important, given the widespread problems associated with drug resistance in many parasitic worms of livestock animals globally.

scholarly journals In vitro activity of Lasioseius penicilliger (Arachnida: Mesostigmata) against three nematode species: Teladorsagia circumcincta, Meloidogyne sp. and Caenorhabditis elegans

2015 ◽  
Vol 2 (1) ◽  
Author(s):  
Noemí García-Ortiz ◽  
Liliana Aguilar-Marcelino ◽  
Pedro Mendoza-de-Gives ◽  
María Eugenia López-Arellano ◽  
Carlos Ramón Bautista-Garfias ◽  
...  
Keyword(s):  
Caenorhabditis Elegans ◽  
Parasitic Nematode ◽  
Biological Control Agent ◽  
Feeding Activity ◽  
Nematode Species ◽  
Predatory Behavior ◽  
Parasitic Nematodes ◽  
Teladorsagia Circumcincta ◽  
Meloidogyne Sp

The aim of this study was to evaluate the predatory behavior in vitro of the mite Lasioseius penicilliger on 3 nematode species: Teladorsagia circumcincta (L3) (a sheep-parasitic nematode), Meloidogyne sp. (J2) (a plant-parasitic nematode), and on various developmental stages of Caenorhabditis elegans (a free-living nematode). The confrontation between mites and nematodes was individually assessed in 2% water agar placed in plastic Petri dishes (2 cm x 1 cm diameter). One thousand nematodes of a species and 5 mites were placed into each plate (10 replicates) and incubated for 5 days at room temperature (18-25ºC). L. penicilliger showed voracious feeding activity against the 3 assessed nematode species. The percentages of predatory activity recorded were 95.1, 80.5 and 79.3 against Meloidogyne sp., C. elegans, and T. circumcincta, respectively (P ≤ 0.05). These results suggest that L. penicilliger has important potential as a biological control agent of parasitic nematodes.

scholarly journals A new strain of Volutella citrinella with nematode predation and nematicidal activity, isolated from the cysts of potato cyst nematodes in China

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Xinyue Zhang ◽  
Hui Zhang ◽  
Zhaochun Jiang ◽  
Qing Bai ◽  
Shishi Wu ◽  
...  
Keyword(s):  
Fermentation Broth ◽  
Bursaphelenchus Xylophilus ◽  
Nematophagous Fungi ◽  
Plant Diseases ◽  
Nematicidal Activity ◽  
Parasitic Nematodes ◽  
Negative Effects ◽  
Cyst Nematodes ◽  
Highly Active

Abstract Background Plant parasitic nematodes (PPNs) are responsible for causing many plant diseases and are extremely difficult to control at present. Currently, due to the negative effects of chemical agents on the environment and human health, the development of new biological pesticides has become an important part of plant nematode control. Nematophagous fungi refers to a class of fungi that kill plant nematodes. Notably, a large number of nematophagous fungi resources remain to be studied. The objective of our study was to use in vitro screening to identify nematophagous fungi and select strains that were highly active against nematodes, providing a primary research for the development and utilization of new nematophagous fungi. Results A new nematophagous fungal strain (GUCC2219) was isolated from cysts of possibly Globodera spp. and Heterodera spp., identified as Volutella citrinella. The hyphae of V. citrinella produced ring structures of variable size and exhibited predatory and nematicidal activity. The hyphal predation rates (in vitro) against three species of nematodes, Aphelenchoides besseyi, Bursaphelenchus xylophilus, and Ditylenchus destructor, averaged 59.45, 33.35, and 50.95%, respectively, while the fermentation broth produced by the fungus exhibited mortality rates of 100, 100, and 55.63%, respectively, after 72 h. Conclusion V. citrinella is a new strain with nematophagous properties, which are a novel discovery. At the same time, this is the first report of nematicidal and nematode predation activity in the genus Volutella.

Sign in / Sign up

Export Citation Format

Share Document