scholarly journals The impact of in vitro cultivation on the natural life cycle of the tick-borne relapsing fever spirochete Borrelia turicatae

PLoS ONE ◽  
2020 ◽  
Vol 15 (10) ◽  
pp. e0239089
Author(s):  
Aparna Krishnavajhala ◽  
Brittany A. Armstrong ◽  
Job E. Lopez
Keyword(s):  
Life Cycle ◽  
In Vitro Cultivation ◽  
Relapsing Fever ◽  
The Impact ◽  
Natural Life

Cloning and Molecular Characterization of a Multicopy, Linear Plasmid-Carried, Repeat Motif-Containing Gene from Borrelia turicatae, a Causative Agent of Relapsing Fever

1998 ◽  
Vol 180 (18) ◽  
pp. 4974-4981 ◽  
Author(s):  
Jason A. Carlyon ◽  
Richard T. Marconi
Keyword(s):  
Transcriptional Start Site ◽  
Gene Families ◽  
In Vitro Cultivation ◽  
Relapsing Fever ◽  
Related Sequence ◽  
Site Analysis ◽  
Low Levels ◽  
Ribosomal Binding Site

ABSTRACT Borrelia turicatae is one of several spirochete species that can cause relapsing fever. Here, we describe the identification and characterization of a gene from B. turicatae and other relapsing-fever spirochetes that exhibits homology with therep + and ORF-E gene families of the Lyme disease spirochetes. This gene, which we have designatedrepA, encodes a putative protein of 30.2 kDa with an isoelectric point of 4.69. The central region of RepA harbors a series of amino acid repeat motifs which exhibit homology with casein kinase 2 phosphorylation sites. Through Southern hybridization analyses, we demonstrate that repA (or a closely related sequence) is multicopy in the relapsing-fever spirochetes and is carried on variably sized linear plasmids in bothBorrelia parkeri and B. turicatae. Transcriptional analyses demonstrate that repA is expressed, albeit at low levels, during in vitro cultivation ofB. turicatae. Transcriptional start site analysis revealed that repA is preceded by a consensus ribosomal binding site and an appropriately spaced promoter element. The sequence conservation, unique features, and multicopy status of repAand its homologs suggest that RepA may play an important genus-wide role in the biology of the Borrelia.

In Vitro Cultivation of Microphallus turgidus (Trematoda: Microphallidae) from Metacercaria to Ovigerous Adult with Continuation of the Life Cycle in the Laboratory

2009 ◽  
Vol 95 (4) ◽  
pp. 913-919 ◽  
Author(s):  
Oscar J. Pung ◽  
Ashley R. Burger ◽  
Michael F. Walker ◽  
Whitney L. Barfield ◽  
Micah H. Lancaster ◽  
...  
Keyword(s):  
Life Cycle ◽  
In Vitro Cultivation

scholarly journals Studies on the Life Cycle of Some New Zealand Anisakidae (Nematoda)

2021 ◽  
Author(s):  
◽  
Rosemary Jennifer Hurst
Keyword(s):  
Life Cycle ◽  
New Zealand ◽  
Body Cavity ◽  
The Body ◽  
In Vitro Cultivation ◽  
Free Living ◽  
Larval Stages ◽  
Factors Influencing ◽  
Phocarctos Hookeri

<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>

scholarly journals Studies on the Life Cycle of Some New Zealand Anisakidae (Nematoda)

2021 ◽  
Author(s):  
◽  
Rosemary Jennifer Hurst
Keyword(s):  
Life Cycle ◽  
New Zealand ◽  
Body Cavity ◽  
The Body ◽  
In Vitro Cultivation ◽  
Free Living ◽  
Larval Stages ◽  
Factors Influencing ◽  
Phocarctos Hookeri

<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>

scholarly journals Molting, Ecdysis, and Reproduction of Trichinella spiralis Are Supported In Vitro by Intestinal Epithelial Cells

2002 ◽  
Vol 70 (4) ◽  
pp. 1853-1859 ◽  
Author(s):  
L. F. Gagliardo ◽  
C. S. McVay ◽  
J. A. Appleton
Keyword(s):  
Life Cycle ◽  
Epithelial Cells ◽  
Intestinal Epithelial Cells ◽  
Trichinella Spiralis ◽  
Mature Male ◽  
In Vitro Cultivation ◽  
Low Density ◽  
Intestinal Epithelial ◽  
Prolonged Culture

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.

scholarly journals Development and optimization of an in vitro cultivation protocol allows for isolation of Borrelia miyamotoi from patients with hard tick-borne relapsing fever

2017 ◽  
Vol 23 (7) ◽  
pp. 480-484 ◽  
Author(s):  
J. Koetsveld ◽  
N.M. Kolyasnikova ◽  
A. Wagemakers ◽  
M.G. Toporkova ◽  
D.S. Sarksyan ◽  
...  
Keyword(s):  
In Vitro Cultivation ◽  
Hard Tick ◽  
Borrelia Miyamotoi ◽  
Relapsing Fever

Methods of intensifying the tuberization process of potato seeds in vitro culture

2017 ◽  
Vol 4 (2) ◽  
pp. 37-41
Author(s):  
H. Balashova ◽  
Yu. Lavrynenko ◽  
R. Vozhehova ◽  
B. Kotov
Keyword(s):  
Experimental Data ◽  
Culture Medium ◽  
In Vitro Cultivation ◽  
Early Potato ◽  
Medium Ph ◽  
In Vitro Methods ◽  
Systemic Analysis ◽  
Effi Ciency ◽  
The Impact

Aim. To determine optimal factors of impacting the tuberization intensity of the plants of mid-early potato variety Nevska in vitro. Methods. Complex application of the laboratory, mathematical-statistical, calculative- comparative methods and systemic analysis. Results. The experimental data on the impact of photoperiod and acidity of the culture medium (pH) on the tuberization induction in culture in vitro were presented. The dependence of tuberization intensity on pH of the culture medium was demonstrated. Conclusions. Optimal indices of tuberization intensity and economic effi ciency were obtained during in vitro cultivation of the plants of mid-early Nevska variety at the photoperiod of 16 hours and pH of the culture medium of 5.3: tuberization intensity was 83.0 %, the mass of the average microtuber was 612.5 mg, the mass of microtubers per one plant was 514.4 mg, the number of microtubers with the mass over 350 mg – 75.4 % with the production profi tability of 100 % and microtuber cost price of UAH 8.08 (USD 0.3).

#85: Nutrient Availability Drives Community Dynamics in the Vaginal Microbiota

2021 ◽  
Vol 10 (Supplement_1) ◽  
pp. S9-S9
Author(s):  
M Indriati Hood-Pishchany ◽  
Seth Rakoff-Nahoum
Keyword(s):  
Community Dynamics ◽  
Bacterial Species ◽  
Nutrient Utilization ◽  
Strain Level ◽  
In Vitro Cultivation ◽  
Carbohydrate Utilization ◽  
Glycogen Utilization ◽  
The Impact

Abstract Background Nutrient utilization is both critical for niche occupation and is the driver of competitive and cooperative interactions in microbial communities. The FRT is replete with host-associated glycans in the form of glycoproteins, epithelial glycogen stores, and the breakdown products of these glycans. I hypothesized that host-associated glycans drive environment, microbe–microbe and host–microbe interactions in the FRT. Methods We have developed robust, scalable, high-throughput culturing systems to empirically define the substrate utilization traits from more than 60 unique bacterial species capable of colonizing the vagina. In addition, we are using batch and continuous culture in vitro cultivation of multispecies communities to study vaginal bacteria within the complex community, that closely recapitulate key dynamics observed in vivo. Results Demonstrating the power of these in vitro models, I have defined the carbohydrate utilization profiles of hundreds of unique FRT isolates, identifying species and strain-level variation in utilization of host-derived carbohydrates. Given the known abundance of glycogen in the vaginal epithelium, I hypothesized that the utilization of host-associated glycogen represents an adaptation to the vaginal environment. Indeed, we identify glycogen degradation enzymes in diverse species resident in the reproductive tract, and find enrichment in genes encoding glycogen-degrading enzymes in L. crispatus strains derived from vaginal as opposed to intestinal sites. Metatranscriptomic analyses from human samples demonstrate that bacterial glycogen and maltose (a breakdown product of glycogen) utilization genes are highly expressed in the vagina and elucidate patterns of gene expression suggestive of context-dependent competition and cooperation for glycogen utilization in vivo. To empirically investigate the impact of glycogen availability and glycogen utilization in FRT microbiota communities, I assembled type strains or co-resident consortia into model, polymicrobial communities in vitro. These studies demonstrate that among health-associated L. crispatus strains, those that use glycogen have a competitive advantage during growth in a complex community. However, preliminary results suggest that some strains may benefit from cross-fed nutrients liberated by other members of the consortium. Conclusions Taken together, these data establish that strain-level variability in glycan utilization contributes to competitive fitness during growth in community, and suggest that these traits may influence community stability or persistence in vivo. Moreover, the methods we have developed provide a scalable system in which to empirically study ecological dynamics within complex community ex vivo.

scholarly journals In Vitro Cultivation of Cryptosporidium Species

2002 ◽  
Vol 15 (3) ◽  
pp. 390-400 ◽  
Author(s):  
Michael J. Arrowood
Keyword(s):  
Life Cycle ◽  
Life Cycle Stage ◽  
In Vitro Cultivation ◽  
Parasite Life Cycle ◽  
Disinfection Methods ◽  
Intracellular Parasites ◽  
Development And Differentiation ◽  
Stage Development ◽  
Life Cycle Development

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.

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