Successful in vitro cultivation of Cryptosporidium andersoni: evidence for the existence of novel extracellular stages in the life cycle and implications for the classification of Cryptosporidium

2002 ◽  
Vol 32 (14) ◽  
pp. 1719-1726 ◽  
Author(s):  
N.S Hijjawi ◽  
B.P Meloni ◽  
U.M Ryan ◽  
M.E Olson ◽  
R.C.A Thompson
2009 ◽  
Vol 95 (4) ◽  
pp. 913-919 ◽  
Author(s):  
Oscar J. Pung ◽  
Ashley R. Burger ◽  
Michael F. Walker ◽  
Whitney L. Barfield ◽  
Micah H. Lancaster ◽  
...  

2021 ◽  
Author(s):  
◽  
Rosemary Jennifer Hurst

<p>The life cycle of Anisakis simplex in New Zealand waters is described from observations on the morphology, distribution and behaviour of free-living and parasitic stages. Comparison with the life cyles of two other anisakids, Phocanema decipiens Myers 1959 and Thynnascaris adunca Rudolphi 1802 shows differences in distribution, degrees of host specificity, the status of invertebrate hosts, the factors influencing infestation levels of teleost hosts, and the location and pathological effects of infestation. Larval stages occurring in intermediate and paratenic hosts were identified by comparison of larval and adult morphometrics. A. simplex larvae were also positively identified by in vitro cultivation through to adults. Some morphometric variations compared to overseas descriptions are apparent. The ventriculus of A. simplex larvae is shorter relative to body length and the intestinal caecum of P. decipiens is longer relative to ventriculus length. Egg and free-living larval stages were obtained from in vitro cultivation of (A. simplex) and collection of eggs from mature adults from definitive hosts (T. adunca). Eggs of P. decipiens were not obtained. Eggs of A. simplex and T. adunca hatch in 8-11 days at 15 [degrees] C. A. simplex eggs hatch in 6 days at a temperature of 22 [degrees] C and did not hatch in 16 days at 10 [degrees] C. Eggs and free-living stage III larvae of A. simplex and T. adunca are similar in morphology with little differentiation of internal structures. Examination of the stomach contents of pelagic fish infested with anisakids indicated that possible intermediate hosts of A. simplex are the euphausiid Nyctiphanes australis and the decapod Munida gregaria. Possible hosts of T. adunca and M. gregaria are a wide variety of smaller zooplanktonic groups, e.g. decapod larvae and copepods. Larvae of A. simplex were found in one of 8850 N. australis; larvae of T. adunca were found in 69 of 3999 chaetognaths (Sagitta spp.) a medusa and a decapod larva. These larvae are morphologically similar to Stage III larvae from teleosts. No anisakids were found in 3956 Euphausia spp., 1147 M. gregaria and 740 prawns. Twenty five T. adunca larvae and adults were found in 818 freshly eaten M. gregaria in teleost stomachs, indicating that this invertebrate may act as a paratenic and a definitive host. Experimental infection of N. australis and M. gregaria with stage II larvae of A. simplex and T. adunca was unsuccessful. The location of anisakid infestation in three pelagic teleost species, Thyrsites atun, Trachurus novaezelandiae and Trachurus declivis is described. A. simplex larvae are found mainly in the body cavity of all species, at the posterior end of the stomach, with less than one percent occurring in the musculature. Distribution of A. simplex larvae does not change with increasing size of the host or increasing total worm burden. Thyrsites atun have a higher proportion of larvae in the stomach wall (8-13%) compared to Trachurus spp. (< 4%). T. adunca larvae are found infrequently in the body cavity of all three species, on the pyloric caeca and in the stomach wall. Adults and larvae of T. adunca are found more commonly in the alimentary canal, indicating that these teleosts are more important as definitive hosts in the life cycle of this anisakid. P. decipiens larvae are found only in Thyrsites atun and occur mainly in the muscles (98.5%). No quantitative pathogenic effects of anisakid infestation on these teleosts hosts were detected. The main factors influencing the infestation of the three teleost species are age of the host, locality and season. Sex of the host and depth (over the continental shelf, 0-250 m) are not important. A. simplex infestation increased with age in all host species examined, and was higher in Trachurus declivis from the southern-most locality, suggesting the existence of at least two distinct populations of this species. Significant differences in infestation of Thyrsites atun with P. decipiens suggests that this anisakid may be more common in southern localities also. The infestation of Thyrsites atun by larval and adult T. adunca in the alimentary canal is most influenced by season and closely related to diet. Nematode samples were obtained from the marine mammals Arctocephalus forsteri, Kogia breviceps and Phocarctos hookeri. Adult A. simplex were recorded from A. forsteri (a new host record) and Kogia breviceps; preadults from Phocarctos hookeri. Adult P. decipiens were recorded from Phocarctos hookeri; preadults from Arctocephalus forsteri and K. breviceps. Other anisakids found were Anisakis physeteris (Baylis 1923), Contracaecum osculatum Rudolphi 1802 and Pseudoterranova kogiae (Johnston and Mawson 1939) Mosgovoi 1951. These records are all new for the New Zealand region except P. decipiens from P. hookeri and C. osculatum from Arctocephalus forsteri. A. simplex and C. osculatum were found associated with gastric ulcers in Arctocephalus forsteri.</p>


2021 ◽  
Author(s):  
◽  
Rosemary Jennifer Hurst

<p>The life cycle of Anisakis simplex in New Zealand waters is described from observations on the morphology, distribution and behaviour of free-living and parasitic stages. Comparison with the life cyles of two other anisakids, Phocanema decipiens Myers 1959 and Thynnascaris adunca Rudolphi 1802 shows differences in distribution, degrees of host specificity, the status of invertebrate hosts, the factors influencing infestation levels of teleost hosts, and the location and pathological effects of infestation. Larval stages occurring in intermediate and paratenic hosts were identified by comparison of larval and adult morphometrics. A. simplex larvae were also positively identified by in vitro cultivation through to adults. Some morphometric variations compared to overseas descriptions are apparent. The ventriculus of A. simplex larvae is shorter relative to body length and the intestinal caecum of P. decipiens is longer relative to ventriculus length. Egg and free-living larval stages were obtained from in vitro cultivation of (A. simplex) and collection of eggs from mature adults from definitive hosts (T. adunca). Eggs of P. decipiens were not obtained. Eggs of A. simplex and T. adunca hatch in 8-11 days at 15 [degrees] C. A. simplex eggs hatch in 6 days at a temperature of 22 [degrees] C and did not hatch in 16 days at 10 [degrees] C. Eggs and free-living stage III larvae of A. simplex and T. adunca are similar in morphology with little differentiation of internal structures. Examination of the stomach contents of pelagic fish infested with anisakids indicated that possible intermediate hosts of A. simplex are the euphausiid Nyctiphanes australis and the decapod Munida gregaria. Possible hosts of T. adunca and M. gregaria are a wide variety of smaller zooplanktonic groups, e.g. decapod larvae and copepods. Larvae of A. simplex were found in one of 8850 N. australis; larvae of T. adunca were found in 69 of 3999 chaetognaths (Sagitta spp.) a medusa and a decapod larva. These larvae are morphologically similar to Stage III larvae from teleosts. No anisakids were found in 3956 Euphausia spp., 1147 M. gregaria and 740 prawns. Twenty five T. adunca larvae and adults were found in 818 freshly eaten M. gregaria in teleost stomachs, indicating that this invertebrate may act as a paratenic and a definitive host. Experimental infection of N. australis and M. gregaria with stage II larvae of A. simplex and T. adunca was unsuccessful. The location of anisakid infestation in three pelagic teleost species, Thyrsites atun, Trachurus novaezelandiae and Trachurus declivis is described. A. simplex larvae are found mainly in the body cavity of all species, at the posterior end of the stomach, with less than one percent occurring in the musculature. Distribution of A. simplex larvae does not change with increasing size of the host or increasing total worm burden. Thyrsites atun have a higher proportion of larvae in the stomach wall (8-13%) compared to Trachurus spp. (< 4%). T. adunca larvae are found infrequently in the body cavity of all three species, on the pyloric caeca and in the stomach wall. Adults and larvae of T. adunca are found more commonly in the alimentary canal, indicating that these teleosts are more important as definitive hosts in the life cycle of this anisakid. P. decipiens larvae are found only in Thyrsites atun and occur mainly in the muscles (98.5%). No quantitative pathogenic effects of anisakid infestation on these teleosts hosts were detected. The main factors influencing the infestation of the three teleost species are age of the host, locality and season. Sex of the host and depth (over the continental shelf, 0-250 m) are not important. A. simplex infestation increased with age in all host species examined, and was higher in Trachurus declivis from the southern-most locality, suggesting the existence of at least two distinct populations of this species. Significant differences in infestation of Thyrsites atun with P. decipiens suggests that this anisakid may be more common in southern localities also. The infestation of Thyrsites atun by larval and adult T. adunca in the alimentary canal is most influenced by season and closely related to diet. Nematode samples were obtained from the marine mammals Arctocephalus forsteri, Kogia breviceps and Phocarctos hookeri. Adult A. simplex were recorded from A. forsteri (a new host record) and Kogia breviceps; preadults from Phocarctos hookeri. Adult P. decipiens were recorded from Phocarctos hookeri; preadults from Arctocephalus forsteri and K. breviceps. Other anisakids found were Anisakis physeteris (Baylis 1923), Contracaecum osculatum Rudolphi 1802 and Pseudoterranova kogiae (Johnston and Mawson 1939) Mosgovoi 1951. These records are all new for the New Zealand region except P. decipiens from P. hookeri and C. osculatum from Arctocephalus forsteri. A. simplex and C. osculatum were found associated with gastric ulcers in Arctocephalus forsteri.</p>


2002 ◽  
Vol 70 (4) ◽  
pp. 1853-1859 ◽  
Author(s):  
L. F. Gagliardo ◽  
C. S. McVay ◽  
J. A. Appleton

ABSTRACT Trichinella spiralis is an obligate parasite of animals that has an unusual intracellular life cycle. Investigation of parasitism at the cellular and molecular levels has been challenging because of a shortage of tools for in vitro cultivation of T. spiralis. We have found that T. spiralis larvae molt, ecdyse, develop to adulthood, and reproduce when they are inoculated onto cultured intestinal epithelial cells. Initially, larvae invade and migrate through cells in a monolayer (T. ManWarren, L. Gagliardo, J. Geyer, C. McVay, S. Pearce-Kelling, and J. Appleton, Infect. Immun. 65:4806-4812, 1997). During prolonged culture in Caco-2 epithelial cells, L1 larvae molted and ecdysed with efficiencies as high as 50%. Molting and ecdysis in vitro required entry of the parasite into cells; conditions that prevented entry into cells also prevented ecdysis. When larvae were inoculated at a low density and cultured for 5 to 9 days, as many as 50% of the larvae developed to adult stages. Low numbers of mature male worms with copulatory appendages were observed in these cultures. The majority of worms that survived for five or more days were unfertilized females. Low-density cultures supported development of female worms with embryos at rates of 4 to 5%. These results show that the intestinal life cycle of T. spiralis can be supported entirely by host epithelial cells. Our model should allow more detailed investigation of intracellular parasitism by T. spiralis.


PLoS ONE ◽  
2020 ◽  
Vol 15 (10) ◽  
pp. e0239089
Author(s):  
Aparna Krishnavajhala ◽  
Brittany A. Armstrong ◽  
Job E. Lopez

2002 ◽  
Vol 15 (3) ◽  
pp. 390-400 ◽  
Author(s):  
Michael J. Arrowood

SUMMARY The in vitro cultivation of protozoan parasites of the genus Cryptosporidium has advanced significantly in recent years. These obligate, intracellular parasites colonize the epithelium of the digestive and respiratory tracts, are often difficult to obtain in significant numbers, produce durable oocysts that defy conventional chemical disinfection methods, and are persistently infectious when stored at refrigerated temperatures (4 to 8°C). While continuous culture and efficient life cycle completion (oocyst production) have not yet been achieved in vitro, routine methods for parasite preparation and cell culture infection and assays for parasite life cycle development have been established. Parasite yields may be limited, but in vitro growth is sufficient to support a variety of research studies, including assessing potential drug therapies, evaluating oocyst disinfection methods, and characterizing life cycle stage development and differentiation.


Author(s):  
Petar Halachev ◽  
Victoria Radeva ◽  
Albena Nikiforova ◽  
Miglena Veneva

This report is dedicated to the role of the web site as an important tool for presenting business on the Internet. Classification of site types has been made in terms of their application in the business and the types of structures in their construction. The Models of the Life Cycle for designing business websites are analyzed and are outlined their strengths and weaknesses. The stages in the design, construction, commissioning, and maintenance of a business website are distinguished and the activities and requirements of each stage are specified.


1996 ◽  
Vol 24 (3) ◽  
pp. 325-331
Author(s):  
Iain F. H. Purchase

The title of this paper is challenging, because the question of how in vitro methods and results contribute to human health risk assessment is rarely considered. The process of risk assessment usually begins with hazard assessment, which provides a description of the inherent toxicological properties of the chemical. The next step is to assess the relevance of this to humans, i.e. the human hazard assessment. Finally, information on exposure is examined, and risk can then be assessed. In vitro methods have a limited, but important, role to play in risk assessment. The results can be used for classification and labelling; these are methods of controlling exposure, analogous to risk assessment, but without considering exposure. The Ames Salmonella test is the only in vitro method which is incorporated into regulations and used widely. Data from this test can, at best, lead to classification of a chemical with regard to genotoxicity, but cannot be used for classification and labelling on their own. Several in vitro test systems which assess the topical irritancy and corrosivity of chemicals have been reasonably well validated, and the results from these tests can be used for classification. The future development of in vitro methods is likely to be slow, as it depends on the development of new concepts and ideas. The in vivo methods which currently have reasonably developed in vitro alternatives will be the easiest to replace. The remaining in vivo methods, which provide toxicological information from repeated chronic dosing, with varied endpoints and by mechanisms which are not understood, will be more difficult to replace.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Malabika Chakrabarti ◽  
Nishant Joshi ◽  
Geeta Kumari ◽  
Preeti Singh ◽  
Rumaisha Shoaib ◽  
...  

AbstractCytoskeletal structures of Apicomplexan parasites are important for parasite replication, motility, invasion to the host cell and survival. Apicortin, an Apicomplexan specific protein appears to be a crucial factor in maintaining stability of the parasite cytoskeletal assemblies. However, the function of apicortin, in terms of interaction with microtubules still remains elusive. Herein, we have attempted to elucidate the function of Plasmodium falciparum apicortin by monitoring its interaction with two main components of parasite microtubular structure, α-tubulin-I and β-tubulin through in silico and in vitro studies. Further, a p25 domain binding generic drug Tamoxifen (TMX), was used to disrupt PfApicortin-tubulin interactions which led to the inhibition in growth and progression of blood stage life cycle of P. falciparum.


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