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J R Barta

Publications and source records attributed to J R Barta.

At least 19 recordsLinked to original sources
PubMed2006

Differential display analysis of gene expression in two immunologically distinct strains of Eimeria maxima.

Gene expression during sporulation and sporozoite excystation of two strains of Eimeria maxima was analyzed using the mRNA differential display technique. The two strains, the Guelph strain (GS) and a single sporocyst-derived strain (M6) from Florida, have been shown to be immunologically distinct. We isolated and cloned a 453-bp complimentary DNA (cDNA) fragment (GS-453) found only in GS sporozoites. In GS, this mRNA begins to be expressed during the earliest stages of oocyst sporulation and is continuously expressed up to and including in the excysted sporozoite. In all Northern blots, digoxigenin (DIG)-labeled GS-453 probe recognized an mRNA of approximately 1.6 kb from GS but not from RNA of M6. Southern blots using various endonucleases and probed with DIG-labeled GS-453 demonstrated that the genomes of both strains contained sufficiently similar sequences to permit hybridization with the probe, but the pattern of hybridization differed between the two strains. Extensive searches of the GenBank, European Molecular Biology Laboratory, and various apicomplexan expressed sequence tag databases using the DNA or inferred amino acid sequences of GS-453 cDNA clone did not identify similarity to any existing sequences.

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PubMed2005

The genus Atoxoplasma (Garnham 1950) as a junior objective synonym of the genus Isospora (Schneider 1881) species infecting birds and resurrection of Cystoisospora (Frenkel 1977) as the correct genus for Isospora species infecting mammals.

Molecular and morphological data permit a rational subdivision of the paraphyletic Isospora into 2 apparently monophyletic groups of parasites, i.e., Isospora and Cystoisospora. Atoxoplasma was determined to be a junior objective synonym for Isospora. Tetrasporozoic, diplosporocystic oocysts possessing Stieda bodies in their sporocysts belong to Isospora (Eimeriidae) and have been described principally from the feces of birds. Tetrasporozoic, diplosporocystic oocysts without Stieda bodies in their sporocysts belong to Cystoisospora (Sarcocystidae).

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PubMed2002

Redescription of Neospora caninum and its differentiation from related coccidia.

Neospora caninum is a protozoan parasite of animals, which before 1984 was misidentified as Toxoplasma gondii. Infection by this parasite is a major cause of abortion in cattle and causes paralysis in dogs. Since the original description of N. caninum in 1988, considerable progress has been made in the understanding of its life cycle, biology, genetics and diagnosis. In this article, the authors redescribe the parasite, distinguish it from related coccidia, and provide accession numbers to its type specimens deposited in museums.

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PubMed2001

Molecular approaches for inferring evolutionary relationships among protistan parasites.

Within the diverse group of parasites broadly recognized as protists, there are limited morphological characters that can be used to distinguish species and even fewer characters that can infer evolutionary relationships among species. For this reason, molecular data are commonly used to infer relationships among species and strains. These studies most commonly rely on sequences associated with the ribosomal RNA genes but increasingly other nuclear, mitochondrial or plastid genes are contributing data. Molecular systematics has been invaluable in expanding the range of characters that are available for inferring relationships among protistan taxa. As an adjunct to morphological characters, sequence data allow us to better understand the evolutionary history of this group of parasites and thereby erect a well-supported taxonomic framework that reflects these historical relationships. Importantly, the predictive nature of such a framework can aid the search for therapeutic compounds (e.g. via shared biochemical pathways) and highlight organisms that should be tested for cross-reactivity in immunological or molecular diagnostic methods (e.g. use of the closest relatives to assess test specificity). For emergent and poorly known parasites, molecular characterization and placement within the broader phylogenetic framework can help predict likely life history traits, including possible or likely definitive hosts.

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PubMed2001

A comparison of sporozoite transport after homologous and heterologous challenge in chickens immunized with the Guelph strain or the Florida strain of Eimeria maxima.

The two strains of Eimeria maxima, Guelph and Florida, used in this study were previously shown to only partially cross-protect immunologically with respect to lesion scores, weight gains and feed conversions after heterologous challenge. In this paper, we provide evidence that this partial lack of cross-protection is manifested at the level of sporozoite transport. In birds immunized and challenged with the homologous strain, sporozoites accumulated in the lamina propria and were blocked from further movement into the crypts by 72 h post-challenge, unlike the situation observed in naive birds. Fewer than 5% of sporozoites were found in the crypts by 72 h post-challenge. In immunized birds challenged with the heterologous strain, fewer sporozoites reached the crypts than in naive birds but at least four times as many sporozoites successfully migrated to the crypts, when compared with birds challenged with the homologous strain. The degree of cross-protection afforded by the heterologous strain as measured by sporozoite transport success was not equally reciprocal.

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PubMed2001

Involvement of CD 8+ and CD 3+ lymphocytes in the transport of Eimeria necatrix sporozoites within the intestinal mucosa of chickens.

The phenotype of cells transporting sporozoites of Eimeria necatrix during a primary infection was determined using a panel of six monoclonal antibodies to various chicken lymphocyte surface markers. Sporozoites and cells harboring them were examined at 8, 12 and 18 h postinfection using two-color immunoflorescence and confocal microscopy. The majority of parasites observed within lymphocytes were found in CD 8 + (15%) or CD 3+ (13-22%) cells at all time periods examined. Smaller numbers were found within deltagamma TCR+ (5%) and alphabeta TCR+ (5%) lymphocytes. No sporozoites were found within CD 4+ or IgM+ lymphocytes at any of the time periods.

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PubMed2001

Chasing coccidia--new tools enter the race.

The 8th International Coccidiosis Conference, held on 9--13 July 2001 in Palm Cove, Australia, was a showcase of the latest studies on widely known coccidia, including Eimeria and Toxoplasma in addition to the emerging or re-emerging parasites such as Neospora, Cryptosporidium and Cyclospora. This meeting was staged in conjunction with the Annual Scientific Meeting of the Australian Society for Parasitology.

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PubMed2001

Molecular phylogeny of the other tissue coccidia: Lankesterella and Caryospora.

Nearly complete sequences were obtained from the 18S rDNA genes of Eimeria falciformis (the type species of the genus), Caryospora bigenetica, and Lankesterella minima. Two clones of the rDNA gene from C. higenetica varied slightly in primary structure. Parsimony-based and maximum likelihood phylogenetic reconstructions with a number of other apicomplexan taxa support 2 major clades within the Eucoccidiorida, i.e., the isosporoid coccidia (consisting of Toxoplasma, Neospora, Isospora [in part], and Sarcocystis spp.) and a second clade containing Lankesterella and Caryospora spp., as well as the eimeriid coccidia (Cyclospora, Isospora [in part], and Eimeria spp.). Our observations suggest that Caryospora spp. may not belong in the family Eimeriidae but rather may be allied with the family Lankesterellidae with which they share molecular and life history similarities. This may be a third lineage of coccidian parasites that has independently evolved a unique heteroxenous transmission strategy.

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PubMed2000

Localization and immunogenicity of a low molecular weight antigen of Eimeria tenella.

A low molecular weight (LMW) antigen recognized by a murine monoclonal antibody (C(3)4F(1)) was localized within endogenous stages of Eimeria tenella (USDA strain 80). Using indirect fluorescent antibody assay and immunoelectron microscopy, the LMW antigen was found in: sporozoites, first, second and third generation meronts, gamonts, unsporulated oocysts, and sporocysts. The antigen was observed in the cytoplasm and pellicle of the parasite, and in the parasitophorous vacuole, sporocyst walls and cytoplasm of infected host cells. The immunogenicity of this LMW antigen was assessed by antigen-specific serum antibody responses in chickens orally inoculated with live oocysts or injected intramuscularly with dead sporozoites. LMW antigen-specific serum antibodies were detected using Western blots of E. tenella sporozoites as early as 4 days after sporozoite injection and 6 days after oocyst inoculation. Unusually, the monoclonal antibody C(3)4F(1) reduced the binding of immune chicken serum to the antigen in a competitive antibody binding assay, but not the reverse, suggesting that there is a single, immunodominant epitope on this antigen.

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PubMed2000

Partial characterization of a non-proteinaceous, low molecular weight antigen of Eimeria tenella.

A low molecular weight (LMW) antigen of Eimeria tenella, initially identified using a murine monoclonal antibody (mAb C(3)4F(1)) raised against E. tenella sporozoites, was partially characterized using enzymatic degradation. solvent extraction, and immunization into various inbred lines of mice. The LMW antigen could be isolated using Folch extraction (methanol/chloroform/ water) and the epitope recognized by mAb C(3)4F(1) was resistant to degradation by alpha-amylase, pronase, and proteinase K, but was sensitive to sodium m-periodate treatment or digestion using mixed glycosidases (from Turbo cornutus). These observations suggest that the antigenic epitope recognized by mAb C(3)4F(1) is carbohydrate-dependent and, based on our ability to isolate the LMW antigen by Folch extraction, the epitope probably resides on a polar glycolipid. The inability of sporozoite-immunized nude mice to elicit a serum antibody response to this molecule indicates that it acts as a T-dependent antigen. Furthermore, sporozoite-immunized male CBA/N mice (with an X-linked immunodeficiency) also failed to elicit a serum antibody response to this molecule, which is consistent with a carbohydrate antigenic epitope. We propose that this antigenic molecule be designated ET-GL1 to reflect its origin and probable structure (E. tenella glycolipid 1).

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PubMed2000

Responses of chickens to a recombinant refractile body antigen of Eimeria tenella administered using various immunizing strategies.

A refractile body antigen (designated SO7') found in sporozoites of Eimeria tenella was administered to chickens in various immunizing forms to assess its ability to protect against virulent challenge. These included native antigen from the parasite (viable oocysts; per os), recombinant antigen (CheY-SO7'; s.c.), and naked DNA immunization (pcDNA3-SO7'; i.m.). Male White Leghorn chicks were inoculated with oocysts or injected with one of the forms of SO7' antigen at 1 wk of age, boosted at 3 wk of age with the same treatment, and subsequently challenged at 5 wk of age with oocysts of Eimeria tenella USDA Strain 80. Seven days postchallenge, chicks were killed, and weight gains and lesion scores were determined and compared with those of the controls. Significant protection against cecal lesions and weight loss was noted in birds inoculated with oocysts or injected with pcDNA3-SO7' (25 microg). Birds injected with other doses of pcDNA3-SO7' (12.5, 50, 60, and 100 microg) showed no reduction in cecal lesions and maintained poor rates of growth similar to controls. The recombinant antigen (CheY-SO7') was also nonprotective. These results offer promise to the use of DNA immunization strategies for controlling avian coccidiosis and show for the first time naked DNA vaccination with a single coccidial antigen that protects chickens (as measured by reduced lesion scores and high rate of growth) against cecal coccidiosis.

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PubMed1999

Cryptosporidium is more closely related to the gregarines than to coccidia as shown by phylogenetic analysis of apicomplexan parasites inferred using small-subunit ribosomal RNA gene sequences.

The phylogenetic placement of gregarine parasites (Apicomplexa: Gregarinasina) within the Apicomplexa was derived by comparison of small-subunit ribosomal RNA gene sequences. Gregarine sequences were obtained from Gregarina niphandrodes Clopton, Percival, and Janovy, 1991, and Monocystis agilis Stein, 1848 (Eugregarinorida Léger 1900), as well as from Ophriocystis elektroscirrha McLaughlin and Myers, 1970 (Neogregarinorida Grassé 1953). The sequences were aligned with several other gregarine and apicomplexan sequences from GenBank and the resulting data matrix analyzed by parsimony and maximum-likelihood methods. The gregarines form a monophyletic clade that is a sister group to Cryptosporidium spp. The gregarine/ Cryptosporidium clade is separate from the other major apicomplexan clade containing the coccidia, adeleids, piroplasms, and haemosporinids. The trees indicate that the genus Cryptosporidium has a closer phylogenetic affinity with the gregarines than with the coccidia. These results do not support the present classification of the Cryptosporidiidae in the suborder Eimerioirina Léger, 1911.

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PubMed1999

An Eimeriid origin of isosporoid coccidia with Stieda bodies as shown by phylogenetic analysis of small subunit ribosomal RNA gene sequences.

Morphological and life cycle features of the tissue cyst-forming coccidia have been difficult to interpret in devising taxonomic classifications for the various genera. In this study, we amplified the full small subunit rRNA gene sequence of Isospora robini McQuistion and Holmes, 1988, and the partial sequence of Isospora gryphoni Olsen, Gissing, Barta, and Middleton, 1998 by PCR. Both of these species vary from Isospora species of mammals in having Stieda bodies on the sporocysts. The sequences were cloned and sequenced and were incorporated into an alignment with other Isospora species lacking Stieda bodies as well as with other coccidia. Maximum parsimony analysis of these sequences produced a single most parsimonious tree that placed I. robini and I. gryphoni in a clade containing various other eimeriid species. The Isospora species lacking Stieda bodies were in the sarcocystid clade. Similar results were found by maximum likelihood analysis. These findings indicate that the genus Isospora as defined by several authors is polyphyletic. Taxonomic changes to the genus Isospora would have to incorporate the 2 major clades found by molecular phylogenetic analysis. Isospora species with Stieda bodies should be classified in the family Eimeriidae, whereas those without Stieda bodies should remain in the family Sarcocystidae.

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PubMed1998

Major histocompatibility complex class I- and II-deficient knock-out mice are resistant to primary but susceptible to secondary Eimeria papillata infections.

Two distinct mechanisms seem to function in reducing oocyst output during Eimeria papillata infections in mice. For naive mice, immunity was afforded by a T-cell-independent gamma-interferon (IFN-gamma) response mediated by natural killer (NK) cells. On reinfection, resistance was associated with T-cells and, to a lesser extent, perforin. To determine if antigen presentation with major histocompatibility complex (MHC) molecules was required to control oocyst production by NK cells during primary infection or by T-cells during secondary infection, mutant mice that lacked H2-IAbeta(b) (Abeta(b)-/-) or beta2-microglobulin (beta2m-/-) were used. Since MHC molecules are required for the maturation of alphabeta T-cells, Abeta(b)-/- and beta2m-/- mutant mice are also deficient in functional alphabeta+CD4+ or alphabeta+CD8+ T-cells, respectively. As compared with wild-type control mice, oocyst output by mutant mice was not significantly affected during primary infection, suggesting that the ability of NK cells to control parasite replication is not dependent on the expression of MHC molecules. On reinfection, differences were observed for mutant mice as compared with controls. Abeta(b)-/- mice were found to be more susceptible than beta2m-/- mice, suggesting that the alphabeta+CD4+ T-cell subset plays a greater role in resistance to reinfection than does the alphabeta+CD8+ T-cell subset. The mechanism of resistance depends on the immune status of the host and requires the coordinated interaction of both alphabeta+ T-cell subsets for optimal parasite control during subsequent infections.

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PubMed1998

Analysis of infraspecific variation among five strains of Eimeria maxima from North America.

Two laboratory strains from the eastern shore of Maryland 15 years ago and from an Ontario broiler house 23 years ago and three recent field strains of Eimeria maxima (isolated in Maryland, North Carolina and Florida) were examined for phenotypic and genotypic variation using protein profiles, random amplified polymorphic DNA-PCR analysis and DNA sequences obtained from the internal transcribed spacer regions of the rRNA genes. Staining profiles obtained by one-dimensional SDS-PAGE of sporozoite proteins were identical in all five strains. Using random amplified polymorphic DNA-PCR analysis with high %G-C content decamers as primers, we were able to confirm that the five strains are all E. maxima, but were unable to discern any relationships among them because of the limited number of shared polymorphisms identified. In contrast, cloning and sequencing of the internal transcribed spacer-1, 5.8S rDNA and internal transcribed spacer-2 regions of the rRNA genes provided sufficient sequence information to infer phylogenetic relationships among the strains. Almost all of the infraspecific variation was located in the internal transcribed spacer regions. Only two base changes were identified within the 5.8S rRNA gene. Evolutionary relationships among the strains inferred using parsimony analysis of the aligned internal transcribed spacer sequences were well supported, but the hypothesised relationships did not correlate well with the demonstrated immunological cross-reactivities of these strains.

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PubMed1998

Plastids are widespread and ancient in parasites of the phylum Apicomplexa.

Current evidence supports the presence of a non-photosynthetic chloroplast-like organelle in several apicomplexan parasites, including Plasmodium falciparum and Toxoplasma gondii. This apicomplexan organelle, referred to here as the "plastid", may have been acquired through a primary or secondary endosymbiosis of a photosynthetic organism. Alternatively, apicomplexan plastids may have been acquired through several independent endosymbiotic events, as appears to be the case for the acquisition of chloroplasts by dinoflagellates. The likelihood of multiple origins of an apicomplexan plastid is enhanced by the close evolutionary relatedness of apicomplexan and dinoflagellate taxa. In this study, we have tested the hypothesis that apicomplexan plastids are derived from a single ancient ancestor. Two lines of evidence supporting this hypothesis are presented. First, this study supports the widespread presence of plastid DNA in apicomplexan species. Second, the topologies of the phylogenetic trees derived from plastid and nuclear-encoded rRNA gene sequences suggest the co-evolution of the DNAs localised in these two compartments. Taken together, these data support a single ancient lineage for the plastids of parasites in the phylum Apicomplexa.

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PubMed1998

Small subunit ribosomal RNA genes of tabanids and hippoboscids (Diptera: Brachycera): evolutionary relationships and comparison with other Diptera.

The small subunit ribosomal RNA (SSU rRNA) genes of hippoboscid (Ornithoica vicina Walker) and tabanid (Chrysops niger Macquart) Diptera were sequenced to determine their phylogenetic position within the order and to determine whether or not extensive hypervariable regions in this gene are widespread in the Diptera. A parsimony analysis of an alignment containing 8 dipteran sequences produced a single most parsimonious tree that placed O. vicina as sister group to Drosophila melanogaster Meigen. The tabanid Chrysops niger was sister group to the asilomorphan taxa, and the sister group to the Brachycera was a Tipula sp. although this relationship was not supported by bootstrap analysis. The hippoboscid and tabanid sequences contain extensive hypervariable regions in the V2, V4, V6, and V7 regions as do other Diptera. When these regions of the alignment were excluded from the phylogenetic analysis, a single most parsimonious tree was found. This tree had an identical overall topology to the tree obtained from the total data set. The hypervariable regions in parts of the dipteran SSU rRNA genes were more extensive in the nematocerous dipteran sequences used in this study than in the other dipteran representatives; these hypervariable regions may be of more utility in inferring relationship among species and subspecies than at the suprageneric level.

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