In vitro development of histotropic larvae of Oesophagostomum dentatum under various conditions of cultivation

Daugschies, Arwid; Watzel, C.

doi:10.1007/s004360050527

Cited by 21 publications

(16 citation statements)

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“…Recent successes with RNAi in parasitic nematodes for selected genes (see Hussein et al 2002;Aboobaker and Blaxter 2004;Lustigman et al 2004;Islam et al 2005;Issa et al 2005) indicate unique prospects for investigating the function of genes directly in O. dentatum. This nematode is considered a unique model for studying the fundamental and developmental biology of parasites (Boag et al 2001(Boag et al , 2003 because it has a rapid and direct life cycle, it produces relatively large numbers of progeny (within a prepatent period of 18-24 days; Talvik et al 1997) and it can also be maintained for longer periods of time in culture in vitro than most bursate nematodes (see Daugschies and Watzel 1999). The results from the present study should provide a basis for discovering the molecular developmental processes associated with UBC-2 in this nematode in vivo and in vitro.…”

Section: Discussionmentioning

confidence: 99%

Ubiquitin-conjugating enzyme genes in Oesophagostomum dentatum

et al. 2006

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Full-length genes representing different isoforms of the ubiquitin-conjugating enzyme UBC-2 were isolated from Oesophagostomum dentatum, cloned and sequenced. The alignment of their sequences (designated Od-ubc-2.1 to Od-ubc-2.3) revealed nucleotide variation at three positions within the predicted open reading frame of 444 bp. Substitutions were at positions 141 (A<-->G), 142 (A<-->G) and 296 (T<-->C). Both former substitutions resulted in amino acid changes from a glycine residue to an arginine residue, whereas the latter resulted in a change from isoleucine to threonine. Comparison of predicted OD-UBC-2 with UBC-2 (protein) homologues/orthologues from 12 other species representing nematodes, Drosophila melanogaster, Saccharomyces cerevisiae, mice and humans revealed identities between species varying from 77 to 100% at the amino acid level, and motifs associated with protein conformation and function were identified. While the function of a representative ubc-2 gene from O. dentatum could not be established in C. elegans, it is likely to play a key role in the catabolism of proteins and in the development of O. dentatum.

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Section: Discussionmentioning

confidence: 99%

Ubiquitin-conjugating enzyme genes in Oesophagostomum dentatum

et al. 2006

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“…(1) the life cycle in vitro cannot be completed under the conditions tested by Daugschies and Watzel (1999), (2) L4 in culture do not reach the size of L4 recovered from pigs (Joachim et al 1999c), (3) larvae do not grow continuously after 25 days cultivation (Experiment 1; Joachim et al 1999c), (4) the size of adults from rectal transplantation of pre-cultured larvae lags behind the size of those from oral infection, and (5) worms recovered ex vivo lose their viability in culture after 1 week.…”

Section: Discussionmentioning

confidence: 99%

“…Adults and L4 recovered from intestinal contents (see below) for cultivation (Experiment 2) were washed intensively in warm 0.9% sodium chloride and transferred carefully to culture¯asks containing either standard larval cultivation medium (Daugschies and Watzel 1999), RPMI-1640 (Life Technologies, Germany) + 10% swine serum, or phosphate-buered saline (PBS; pH 7.2) + 10% swine serum, including double the amount of antibiotics described (Daugschies and Watzel 1999). In these cultures (c. 150 L4 or 50 adults/2.5 ml), the medium was changed daily for the ®rst 3 days and every 3 days thereafter.…”

Section: Cultivation Of Parasites and Recovery From Intestinal Contentsmentioning

confidence: 99%

“…For the cultivation of larvae, sheathed L3 recovered from the faeces and puri®ed by small-scale agar-gel migration (Daugschies 1995) were exsheathed and incubated in standard cultivation medium in densities of 10,000 L3/2.5 ml medium as described by Daugschies and Watzel (1999) for 14, 21 or 26 days with weekly changes of medium. L4 were separated from L3 by sedimentation (Joachim et al 1998) and kept at 37°C in 0.9% sodium chloride until further use (<1 h).…”

Section: Cultivation Of Parasites and Recovery From Intestinal Contentsmentioning

confidence: 99%

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Comparative studies on the development of Oesophagostomum dentatum in vitro and in vivo

Joachim

Ruttkowski²,

Daugschies³

2001

Parasitology Research

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Although in vitro cultivation of Oesophagostomum dentatum provides defined material of third- (L3) and fourth- (L4) stage larvae, these are morphologically and biochemically different from larvae recovered ex vivo. The development of pre-cultivated larvae was investigated by rectal transplantation into worm-free pigs with subsequent recovery of worms from intestinal contents after different time periods and determination of worm burdens and sizes. Additionally, the in vitro maintenance of L4 and adults recovered from intestinal contents of orally infected pigs in different media was investigated. Although growth and development rates of cultivated L4 are lower than those of larvae recovered from intestinal contents after oral infection, pre-cultured L4 are able to develop into egg-laying adults in the large intestines (without nodule formation) within 9-14 days in rates comparable with those after oral infection. In contrast, rectally transplanted L3 only establish in low numbers without egg excretion. L4 and adult worms recovered from intestinal contents cannot be maintained in cultivation medium for more than 1 week, although most L4 grow and moult during the first 3 days. Although the standard cultivation conditions for mass production of L4 are not suitable for development or maintenance of preadult and adult stages, L4 recovered from cultures have the ability to establish in vivo as fertile adults, indicating that the basic biological functions are retained in vitro.

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“…Several studies conducted on in vitro culture/survival of the parasitic helminthes were of Hall 1971;Narien et al 1995;Narien 1980;Kuchler 1928;Weinland 1901;Weinstain and Jones 1959;Tayler and Baker 1968;Daugschies and Watzel 1999;Hata 1978 andHata et al 1980. In vitro culture of nematodes parasites is important because of their medical, veterinary, and economic importance, extensive work has been carried out on the Ostertagia could never live for more than 24 h in sodium chloride solution and with addition of (0.23) percent calcium chloride solution the above parasite could live for 3 days (Davey 1938).…”

Section: Introductionmentioning

confidence: 99%

A study on in vitro culture of Trichuris ovis in different physiological solutions at constant temperature, 37°C

Singh

Lal

2011

J Parasit Dis

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The primary aim of in vitro culture of whipworms (Trichuris ovis) is to provide artificial conditions under which the life cycle of the parasites completed outside the host under controlled conditions. The physiological solutions used for the present study were sodium chloride (0.64%), Ringer's solution, Tyrode's solution, and Lock-Lewis solution. Parasites were collected from freshly slaughtered intestine of the host. The recovered parasites were washed with running tap water after that with normal saline. After washing parasites were put in four petridishes containing different physiological solutions. Observations were recorded after interval of every 8 h. The hundred percent survival of Trichuris ovis was observed at 32, 40, and 48 h in NaCl (0.64%), Ringer's, Tyrode's, and LockLewis solution, respectively in case of both male and female parasites. In sodium chloride solution (0.64%) cent percent mortality was observed after 64 h of incubation in males and in case of females it was observed 72 h. In Ringer's solution cent percent mortality was observed after 72 in males and in females it was observed 80 h. In Tyrode's solution cent percent mortality was observed after 88 h in males and 96 h in females. In Lock-Lewis solution cent percent mortality was observed after 96 h in case of both the male and female parasites. Present study could be used to understand the effects of various drugs on the above parasites and also other intra-intestinal parasites.

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In vitro development of histotropic larvae of Oesophagostomum dentatum under various conditions of cultivation

Cited by 21 publications

References 6 publications

Ubiquitin-conjugating enzyme genes in Oesophagostomum dentatum

Ubiquitin-conjugating enzyme genes in Oesophagostomum dentatum

Comparative studies on the development of Oesophagostomum dentatum in vitro and in vivo

A study on in vitro culture of Trichuris ovis in different physiological solutions at constant temperature, 37°C

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