Optimization of the Agar-gel Method for Isolation of Migrating Ascaris suumLarvae From the Liver and Lungs of Pigs

Saeed, I.; Roepstorff, Allan; Rasmussen, Torben Valdbjørn; Høg, M.; Jungersen, Gregers

doi:10.1186/1751-0147-42-279

Cited by 7 publications

(1 citation statement)

References 14 publications

(18 reference statements)

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“…Pigs were sacrificed at 8 days post-infection for lung-stage larvae, and 16 days post-infection for L4-stage larvae. Lung-stage L3 larvae were retrieved as previously described with minor modifications (Slotved et al, 1997 ; Saeed et al, 2001 ). Briefly, harvested organs were ground using a hand-operated meat grinder.…”

Section: Methodsmentioning

confidence: 99%

The Intestinal Roundworm Ascaris suum Releases Antimicrobial Factors Which Interfere With Bacterial Growth and Biofilm Formation

Midha

Janek

Niewienda

et al. 2018

Front. Cell. Infect. Microbiol.

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Ascariasis is a widespread soil-transmitted helminth infection caused by the intestinal roundworm Ascaris lumbricoides in humans, and the closely related Ascaris suum in pigs. Progress has been made in understanding interactions between helminths and host immune cells, but less is known concerning the interactions of parasitic nematodes and the host microbiota. As the host microbiota represents the direct environment for intestinal helminths and thus a considerable challenge, we studied nematode products, including excretory-secretory products (ESP) and body fluid (BF), of A. suum to determine their antimicrobial activities. Antimicrobial activities against gram-positive and gram-negative bacterial strains were assessed by the radial diffusion assay, while effects on biofilm formation were assessed using the crystal violet static biofilm and macrocolony assays. In addition, bacterial neutralizing activity was studied by an agglutination assay. ESP from different A. suum life stages (in vitro-hatched L3, lung-stage L3, L4, and adult) as well as BF from adult males were analyzed by mass spectrometry. Several proteins and peptides with known and predicted roles in nematode immune defense were detected in ESP and BF samples, including members of A. suum antibacterial factors (ASABF) and cecropin antimicrobial peptide families, glycosyl hydrolase enzymes such as lysozyme, as well as c-type lectin domain-containing proteins. Native, unconcentrated nematode products from intestine-dwelling L4-stage larvae and adults displayed broad-spectrum antibacterial activity. Additionally, adult A. suum ESP interfered with biofilm formation by Escherichia coli, and caused bacterial agglutination. These results indicate that A. suum uses a variety of factors with broad-spectrum antibacterial activity to affirm itself within its microbe-rich environment in the gut.

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Section: Methodsmentioning

confidence: 99%

The Intestinal Roundworm Ascaris suum Releases Antimicrobial Factors Which Interfere With Bacterial Growth and Biofilm Formation

Midha

Janek

Niewienda

et al. 2018

Front. Cell. Infect. Microbiol.

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Ascaridia galli in chickens: intestinal localization and comparison of methods to isolate the larvae within the first week of infection

et al. 2012

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This study was conducted to observe the localization and to compare methods for isolation of minute Ascaridia galli larvae in chicken intestine. Firstly, six 7-week-old layer pullets were orally infected with 2,000 embryonated A. galli eggs and necropsied either at 3, 5 or 7 days post infection (dpi). More than 95 % of the recovered larvae were obtained from the anterior half of the jejunoileum, suggesting this part as the initial predilection site for A. galli larvae. Secondly, the intestinal wall of one layer pullet infected with 20,000 A. galli eggs 3 days earlier was digested in pepsin-HCl for 90 min. The initial 10 min of digestion released 51 % of the totally recovered larvae and the last 30 min of continuous digestion yielded only 5 %. This indicates that the majority of larvae were located superficially in the intestinal mucosa. Thirdly, 48 7-week-old layer pullets were infected with 500 A. galli eggs and necropsied at 3 dpi to compare three different larval isolation methods from the intestinal wall, viz., EDTA incubation, agar-gel incubation and pepsin-HCl digestion, resulting in mean percentages of the recovered larvae: 14.4, 18.2 and 20.0 %, respectively (P = 0.15). As conclusion, we recommended Pepsin-HCl digestion as the method of choice for larval recovery from the intestinal wall in future population dynamics study due to high efficiency and quick and simple detection. The agar-gel method was considered to be a prerequisite for molecular and immunological investigations as the larvae were more active and fully intact.

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WITHDRAWN: Co-administration of CCL5 and IL4 enhances DNA vaccine-induced protective immunity to Ascaris suum infection in mice

Luo

Wang

Zhangwe

et al. 2013

International Journal for Parasitology

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Optimization of the Agar-gel Method for Isolation of Migrating Ascaris suumLarvae From the Liver and Lungs of Pigs

Cited by 7 publications

References 14 publications

The Intestinal Roundworm Ascaris suum Releases Antimicrobial Factors Which Interfere With Bacterial Growth and Biofilm Formation

The Intestinal Roundworm Ascaris suum Releases Antimicrobial Factors Which Interfere With Bacterial Growth and Biofilm Formation

Ascaridia galli in chickens: intestinal localization and comparison of methods to isolate the larvae within the first week of infection

WITHDRAWN: Co-administration of CCL5 and IL4 enhances DNA vaccine-induced protective immunity to Ascaris suum infection in mice

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