Hatching and in Vitro Cultivation of the Nematode Ascaridia galli to the Third-Stage Larva

Dick, J. W.; Leland, S. E.; Hansen, M. F.

doi:10.2307/3224919

Cited by 7 publications

(14 citation statements)

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“…At the end of each incubation period at 26°C, embryonated eggs were subjected to in vitro hatching, liberating the larvae using a modification of the method developed by Dick et al . (1973). Ascaridia galli eggs were placed in a solution containing equal parts of 4% NaHO (Chemsupply Pty Ltd, Gillman, Australia) and 4% NaClO (Pental Limited, Shepparton, Australia) for 24 h at 25°C.…”

Section: Methodsmentioning

confidence: 99%

“…It has been suggested that not all eggs obtained in uteri are mature and able to complete embryonation (Tiersch et al ., 2013) whereas adult female worms appear to shed only mature eggs (Kim et al ., 2012). In line with this, a number of A. galli studies have used eggs oviposited by mature female worms under in vitro incubation in artificial media such as physiological saline (0.85% NaCl) (Dick et al ., 1973; Salih & Saleem, 1987) and Roswell Park Memorial Institute (RPMI) media (Ruhnke et al ., 2017; Sharma et al ., 2017). This approach can be considered the most feasible, efficient and established method of recovering mature A. galli eggs for experimental purposes.…”

Section: Introductionmentioning

confidence: 99%

“…Early studies showed that the infectivity (Elliott, 1954) and severity of infection (Todd et al ., 1952) of A. galli larvae diminished with age of embryonated eggs. Eggs used to induce A. galli infection can be obtained from several sources: the host excreta (Luna-Olivares et al ., 2012; Ferdushy et al ., 2013), physical removal from, or disruption of, the worm's uterus (Gauly et al ., 2002; Daş et al ., 2010) or by in vitro culturing of female worms in artificial media and recovering eggs shed into the media typically across 3–5 days (Dick et al ., 1973; Salih & Saleem, 1987; Ruhnke et al ., 2017; Sharma et al ., 2017).…”

Section: Introductionmentioning

confidence: 99%

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Viability and development of Ascaridia galli eggs recovered in artificial media followed by storage under different conditions

Feyera

Ruhnke

Sharpe³

et al. 2020

J. Helminthol.

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Eggs oviposited by Ascaridia galli females in artificial media are commonly used as a source of infective material. We investigated the rate of egg production by cultured mature females (n = 223), and changes in egg viability under different storage and incubation conditions. Eggs recovered after 1, 2 or 3 days of culture were subjected to either (1) storage in water at 4°C (1, 4 or 8 weeks) followed by incubation in 0.1 N H2SO4 at 26°C (2, 4 or 6 weeks); or (2) prolonged storage at 4°C (up to 14 weeks). Egg development and viability was assessed by morphology coupled with a viability dye exclusion test of hatched larvae. Of the 6,044 eggs recovered per mature female 49.2, 38.5 and 12.3% were recovered on days 1, 2 and 3 of worm incubation respectively with similar initial viability (≥99%) between days. Eggs recovered on different days had only minor differences in viability after storage. The prolonged storage period at 4°C significantly affected both viability and embryonation ability resulting in decline in viability of 5.7–6.2% per week. A smaller but significant decline in egg (2.0%) and hatched larval (1.4%) viability per week of incubation at 26°C was also observed. We conclude that storage and incubation conditions, not the day of egg recovery, are the main factors affecting A. galli egg viability. Our findings indicate that under aerobic conditions storage at 26°C may be preferable to 4°C whereas other studies indicate that under anaerobic conditions storage at 4°C is preferable.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Viability and development of Ascaridia galli eggs recovered in artificial media followed by storage under different conditions

Feyera

Ruhnke

Sharpe³

et al. 2020

J. Helminthol.

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“…After ingestion, the infective eggs are mechanically transported to the proventricalus and gizzard and further down to the duodenum where they hatch within the first 24 hours. Triggering factors that signal the larvae to hatch are believed to be temperature, carbon dioxide level and pH levels (Dick et al 1973;Salih and Saleem 1987). Following hatching, the larvae burrow into the mucosal layer of the small intestine to enter the histotrophic phase (Ackert 1931).…”

Section: Life Cyclementioning

confidence: 99%

Environmental tolerance of free-living stages of the poultry roundworm Ascaridia galli

Tarbiat

Jansson

Höglund

2015

Veterinary Parasitology

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“…As embryonation of nematode eggs is affected by several environmental conditions (Dick et al 1973; Anderson 1992; Permin et al 1997a) and shows species-specific characteristics, specific requirements must be determined. Two important factors influencing embryonation and the subsequent infectivity of nematode eggs are culture media and duration of incubation.…”

Section: Introductionmentioning

confidence: 99%

The role of culture media on embryonation and subsequent infectivity of Capillaria obsignata eggs

et al. 2012

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This study investigated whether infectivity of Capillaria obsignata eggs depends on media culture used for embryonation. Intact female worms were kept in one of following four media: 0.5 % formalin, 2 % formalin, 0.1 % potassium dichromate and 0.1 N sulfuric acid. Embryonation rates of the eggs were quantified either daily in intact females for 16 days, or weekly in disrupted females. Infectivity of the embryonated eggs was tested through an experimental infection of chickens with a single dose of 250 eggs/ bird. The vast majority of the eggs (>82 %) in the first two thirds of the uteri was able to complete embryonation, irrespective of the culture media used for incubation. However, only 32.6 % of total eggs could be harvested after disruption of the intact females. Embryonation rates of the eggs from disrupted worms were different among four culture media, with 0.1 N sulfuric acid resulting in the highest embryonation rate (44.2 %). All the experimentally infected birds harboured mature worms, with varying establishment rates depending on the culture media (P < 0.001). Incubation of the eggs in potassium dichromate 0.1 % resulted in a lower (P < 0.001) establishment rate (10.2 %) when compared with formalin (70.5 and 47.9 % for concentrations at 0.5 and 2 %, respectively) or with 0.1 N sulfuric acid (57.5 %). It can be concluded that most of the eggs in first two thirds of the uteri in the intact females have the potential to complete embryonation without being influenced by the culture media. However, disruption of the intact females results in lower number of harvestable embryonated eggs, with a considerable variation due to culture media used. With the exception of 0.1 % potassium dichromate, any of the three media, particularly 0.1 N sulfuric acid, can be suggested for embryonation of C. obsignata eggs.

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Hatching and in Vitro Cultivation of the Nematode Ascaridia galli to the Third-Stage Larva

Cited by 7 publications

References 0 publications

Viability and development of Ascaridia galli eggs recovered in artificial media followed by storage under different conditions

Viability and development of Ascaridia galli eggs recovered in artificial media followed by storage under different conditions

Environmental tolerance of free-living stages of the poultry roundworm Ascaridia galli

The role of culture media on embryonation and subsequent infectivity of Capillaria obsignata eggs

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