scholarly journals Culturing and Characterization of Gut Symbiont Burkholderia spp. from the Southern Chinch Bug, Blissus insularis (Hemiptera: Blissidae)

2016 ◽  
Vol 82 (11) ◽  
pp. 3319-3330 ◽  
Author(s):  
Yao Xu ◽  
Eileen A. Buss ◽  
Drion G. Boucias
Keyword(s):  
Organ Culture ◽  
Genetic Manipulation ◽  
Culture Method ◽  
Content Type ◽  
Chinch Bug ◽  
Midgut Crypts ◽  
Blissus Insularis ◽  
Genotypic Characteristics ◽  
Southern Chinch Bug

ABSTRACTThe phloem-feeding Southern chinch bug,Blissus insularis, harbors a high density of the exocellular bacterial symbiontBurkholderiain the lumen of specialized midgut crypts. Here we developed an organ culture method that initially involved incubating theB. insulariscrypts in osmotically balanced insect cell culture medium. This approach enabled the crypt-inhabitingBurkholderiaspp. to make a transition to anin vitroenvironment and to be subsequently cultured in standard bacteriological media. Examinations using ribotyping and BOX-PCR fingerprinting techniques demonstrated that mostin vitro-produced bacterial cultures were identical to their crypt-inhabitingBurkholderiacounterparts. Genomic and physiological analyses of gut-symbioticBurkholderiaspp. that were isolated individually from two separateB. insularislaboratory colonies revealed that the majority of individual insects harbored a singleBurkholderiaribotype in their midgut crypts, resulting in a diverseBurkholderiacommunity within each colony. The diversity was also exhibited by the phenotypic and genotypic characteristics of theseBurkholderiacultures. Access to cultures of crypt-inhabiting bacteria provides an opportunity to investigate the interaction between symbioticBurkholderiaspp. and theB. insularishost. Furthermore, the culturing method provides an alternative strategy for establishingin vitrocultures of other fastidious insect-associated bacterial symbionts.IMPORTANCEAn organ culture method was developed to establishin vitrocultures of a fastidiousBurkholderiasymbiont associated with the midgut crypts of the Southern chinch bug,Blissus insularis. The identities of the resulting cultures were confirmed using the genomic and physiological features ofBurkholderiacultures isolated fromB. insulariscrypts, showing that host insects maintained the diversity ofBurkholderiaspp. over multiple generations. The availability of characterized gut-symbioticBurkholderiacultures provides a resource for genetic manipulation of these bacteria and for examination of the mechanisms underlying insect-bacterium symbiosis.

scholarly journals Variation in southern chinch bug ( Blissus insularis ) survival and damage on St. Augustinegrass germplasm

itsrj ◽  
2021 ◽  
Author(s):  
Rocio Laat ◽  
Adam G. Dale ◽  
Consuelo Arellano ◽  
Susana R. Milla‐Lewis
Keyword(s):  
Chinch Bug ◽  
Blissus Insularis ◽  
Southern Chinch Bug

scholarly journals In vitro mouse spermatogenesis with an organ culture method in chemically defined medium

PLoS ONE ◽  
2018 ◽  
Vol 13 (2) ◽  
pp. e0192884 ◽  
Author(s):  
Hiroyuki Sanjo ◽  
Mitsuru Komeya ◽  
Takuya Sato ◽  
Takeru Abe ◽  
Kumiko Katagiri ◽  
...  
Keyword(s):  
Organ Culture ◽  
Culture Method ◽  
Defined Medium ◽  
Chemically Defined Medium

scholarly journals Nonselective Persistence of a Rickettsia conorii Extrachromosomal Plasmid during Mammalian Infection

2016 ◽  
Vol 84 (3) ◽  
pp. 790-797 ◽  
Author(s):  
Sean P. Riley ◽  
Abigail I. Fish ◽  
Daniel A. Garza ◽  
Kaikhushroo H. Banajee ◽  
Emma K. Harris ◽  
...  
Keyword(s):  
Pathogenic Bacteria ◽  
Genetic Manipulation ◽  
Fluorescent Proteins ◽  
Spotted Fever ◽  
Rickettsia Conorii ◽  
Mediterranean Spotted Fever ◽  
Extrachromosomal Dna ◽  
Content Type

Scientific analysis of the genusRickettsiais undergoing a rapid period of change with the emergence of viable genetic tools. The development of these tools for the mutagenesis of pathogenic bacteria will permit forward genetic analysis ofRickettsiapathogenesis. Despite these advances, uncertainty still remains regarding the use of plasmids to study these bacteria inin vivomammalian models of infection, namely, the potential for virulence changes associated with the presence of extrachromosomal DNA and nonselective persistence of plasmids in mammalian models of infection. Here, we describe the transformation ofRickettsia conoriiMalish 7 with the plasmid pRam18dRGA[AmTrCh]. TransformedR. conoriistably maintains this plasmid in infected cell cultures, expresses the encoded fluorescent proteins, and exhibits growth kinetics in cell culture similar to those of nontransformedR. conorii. Using a well-established murine model of fatal Mediterranean spotted fever, we demonstrate thatR. conorii(pRam18dRGA[AmTrCh]) elicits the same fatal outcomes in animals as its untransformed counterpart and, importantly, maintains the plasmid throughout infection in the absence of selective antibiotic pressure. Interestingly, plasmid-transformedR. conoriiwas readily observed both in endothelial cells and within circulating leukocytes. Together, our data demonstrate that the presence of an extrachromosomal DNA element in a pathogenic rickettsial species does not affect eitherin vitroproliferation orin vivoinfectivity in models of disease and that plasmids such as pRam18dRGA[AmTrCh] are valuable tools for the further genetic manipulation of pathogenic rickettsiae.

Interrelationship of Big-Eyed Bugs (Hemiptera: Lygaeidae) and Southern Chinch Bugs (Hemiptera: Lygaeidae) in Florida Lawns2

2005 ◽  
Vol 40 (4) ◽  
pp. 385-389
Author(s):  
Ron Cherry
Keyword(s):  
Significant Positive Correlation ◽  
Insect Pest ◽  
Stenotaphrum Secundatum ◽  
Positive Correlation ◽  
Chinch Bug ◽  
Predator Response ◽  
Blissus Insularis ◽  
Southern Chinch Bug

The southern chinch bug, Blissus insularis Barber, is the most important insect pest of St. Augustinegrass, Stenotaphrum secundatum (Walt.) Kuntze. Fifteen chinch bug infestations in St. Augustinegrass lawns in Florida were sampled by vacuuming. Additional vacuum samples were taken in 15 randomly selected St. Augustinegrass lawns. The big-eyed bug, Geocoris uliginosus Say, was the most frequent and abundant big-eyed bug found at the infestations. Data showed that big-eyed bugs (Geocoris spp.) were highly aggregated at chinch bug infestations. Furthermore, there was a significant positive correlation between numbers of chinch bugs and big-eyed bugs at chinch bug infestations showing that big-eyed bugs had a numerical predator response to increasing chinch bug populations.

scholarly journals Altered Murine Tissue Colonization by Borrelia burgdorferi following Targeted Deletion of Linear Plasmid 17-Carried Genes

2012 ◽  
Vol 80 (5) ◽  
pp. 1773-1782 ◽  
Author(s):  
Timothy Casselli ◽  
Yvonne Tourand ◽  
Troy Bankhead
Keyword(s):  
Lyme Disease ◽  
Borrelia Burgdorferi ◽  
Mutant Strain ◽  
Genetic Manipulation ◽  
Infectious Clone ◽  
Linear Plasmid ◽  
Peripheral Tissues ◽  
Content Type ◽  
Immunodeficient Mice

ABSTRACTThe causative agent of Lyme disease,Borrelia burgdorferi, possesses a segmented genome comprised of a single linear chromosome and upwards of 23 linear and circular plasmids. Much of what is known about plasmid-borne genes comes from studying laboratory clones that have spontaneously lost one or more plasmids duringin vitropassage. Some plasmids, including the linear plasmid lp17, are never or rarely reported to be lost during routine culture; therefore, little is known about the requirement of these conserved plasmids for infectivity. In this study, the effects of deleting regions of lp17 were examined bothin vitroandin vivo. A mutant strain lacking the genesbbd16tobbd25showed no deficiency in the ability to establish infection or disseminate to the bloodstream of mice; however, colonization of peripheral tissues was delayed. Despite the ability to colonize ear, heart, and joint tissues, this mutant exhibited a defect in bladder tissue colonization for up to 56 days postinfection. This phenotype was not observed in immunodeficient mice, suggesting that bladder colonization by the mutant strain was inhibited by an adaptive immune-based mechanism. Moreover, the mutant displayed increased expression of outer surface protein Cin vitro, which was correlated with the absence of the genebbd18. To our knowledge, this is the first report involving genetic manipulation of lp17 in an infectious clone ofB. burgdorferiand reveals for the first time the effects of lp17 gene deletion during murine infection by the Lyme disease spirochete.

Production of collagenase and inhibitor (TIMP) by intracranial tumors and dura in vitro

1983 ◽  
Vol 59 (3) ◽  
pp. 461-466 ◽  
Author(s):  
Ahmed N. Halaka ◽  
Rowena A.D. Bunning ◽  
Colin C. Bird ◽  
Myles Gibson ◽  
John J. Reynolds
Keyword(s):  
Organ Culture ◽  
Tumor Invasion ◽  
Culture Media ◽  
Intracranial Tumors ◽  
Short Term ◽  
Content Type ◽  
Collagenase Inhibitor ◽  
Tissue Degradation ◽  
Fine Print

✓ The production of collagenase and collagenase inhibitor (TIMP) by various intracranial tumors (25 meningiomas, eight gliomas, seven metastases, four pituitary adenomas, and five others) was studied in short-term organ culture. While meningiomas produced negligible amounts of collagenase, two metastatic carcinomas of bronchial and breast origin produced significant amounts of the enzyme. Cultures of dura from an invasive meningioma and of bone invaded by a meningioma also produced collagenase. In varying amounts, TIMP was detected in culture media from most of the tumors studied; invasive tumors tended to produce less TIMP than noninvasive tumors. The results are discussed in relation to current views on tissue degradation and mechanisms of tumor invasion.

scholarly journals Simulating the evolutionary trajectories of metabolic pathways for insect symbionts in the genus Sodalis

2020 ◽  
Vol 6 (7) ◽  
Author(s):  
Rebecca J. Hall ◽  
Stephen Thorpe ◽  
Gavin H. Thomas ◽  
A. Jamie Wood
Keyword(s):  
Type Species ◽  
Genetic Manipulation ◽  
Balance Model ◽  
Ecological Change ◽  
Content Type ◽  
Link Type ◽  
Starting Point ◽  
Evolutionary Trajectories ◽  
Insect Symbionts

Insect–bacterial symbioses are ubiquitous, but there is still much to uncover about how these relationships establish, persist and evolve. The tsetse endosymbiont Sodalis glossinidius displays intriguing metabolic adaptations to its microenvironment, but the process by which this relationship evolved remains to be elucidated. The recent chance discovery of the free-living species of the genus Sodalis , Sodalis praecaptivus , provides a serendipitous starting point from which to investigate the evolution of this symbiosis. Here, we present a flux balance model for S. praecaptivus and empirically verify its predictions. Metabolic modelling is used in combination with a multi-objective evolutionary algorithm to explore the trajectories that S. glossinidius may have undertaken from this starting point after becoming internalized. The order in which key genes are lost is shown to influence the evolved populations, providing possible targets for future in vitro genetic manipulation. This method provides a detailed perspective on possible evolutionary trajectories for S. glossinidius in this fundamental process of evolutionary and ecological change.

Southern Chinch Bug, Blissus insularis Barber (Hemiptera: Blissidae)

2008 ◽  
pp. 3470-3470
Author(s):  
John B. Heppner ◽  
David B. Richman ◽  
Steven E. Naranjo ◽  
Dale Habeck ◽  
Christopher Asaro ◽  
...  
Keyword(s):  
Chinch Bug ◽  
Blissus Insularis ◽  
Southern Chinch Bug

Survival of Different Life Stages of the Southern Chinch Bug (Hemiptera: Lygaeidae) Following Insecticidal Applications

1999 ◽  
Vol 34 (1) ◽  
pp. 126-131 ◽  
Author(s):  
Russell T. Nagata ◽  
Ron H. Cherry
Keyword(s):  
Life Stages ◽  
Stenotaphrum Secundatum ◽  
Chinch Bug ◽  
Lambda Cyhalothrin ◽  
Topical Treatments ◽  
Blissus Insularis ◽  
Southern Chinch Bug

Survival of different life stages of the southern chinch bug, Blissus insularis Barber, was measured after insecticidal applications of acephate, chlorpyrifos, and lambda-cyhalothrin. Adults and nymphs, but not eggs, were killed with all three insecticides sprayed at recommended field rates. Even when sprigs of St. Augustinegrass, Stenotaphrum secundatum (Walt.) Kuntze, were sprayed to runoff (drench), eggs were not killed. In topical treatments, only chlorpyrifos killed eggs when the insecticides were applied directly to the eggs.

scholarly journals Cryptosporidium Pathogenicity and Virulence

2013 ◽  
Vol 26 (1) ◽  
pp. 115-134 ◽  
Author(s):  
Maha Bouzid ◽  
Paul R. Hunter ◽  
Rachel M. Chalmers ◽  
Kevin M. Tyler
Keyword(s):  
Virulence Factors ◽  
Genetic Manipulation ◽  
Protozoan Parasite ◽  
Molecular Techniques ◽  
Host Susceptibility ◽  
Content Type ◽  
Susceptibility To Infection ◽  
Substantial Progress ◽  
Made In

SUMMARYCryptosporidiumis a protozoan parasite of medical and veterinary importance that causes gastroenteritis in a variety of vertebrate hosts. Several studies have reported different degrees of pathogenicity and virulence amongCryptosporidiumspecies and isolates of the same species as well as evidence of variation in host susceptibility to infection. The identification and validation ofCryptosporidiumvirulence factors have been hindered by the renowned difficulties pertaining to thein vitroculture and genetic manipulation of this parasite. Nevertheless, substantial progress has been made in identifying putative virulence factors forCryptosporidium. This progress has been accelerated since the publication of theCryptosporidium parvumandC. hominisgenomes, with the characterization of over 25 putative virulence factors identified by using a variety of immunological and molecular techniques and which are proposed to be involved in aspects of host-pathogen interactions from adhesion and locomotion to invasion and proliferation. Progress has also been made in the contribution of host factors that are associated with variations in both the severity and risk of infection. Here we provide a review comprised of the current state of knowledge onCryptosporidiuminfectivity, pathogenesis, and transmissibility in light of our contemporary understanding of microbial virulence.

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