Biomaterial Database

Individual oocysts of Isospora (Apicomplexa: Coccidia) parasites from avian feces: from photo to sequence. 2009

Olga V Dolnik, and Vaidas Palinauskas, and Staffan Bensch

Department of Animal Ecology, Lund University, Sölvegatan 37, 22362 Lund, Sweden. odolnik@ipoe.uni-kiel.de

Numerous microscopic studies of coccidian oocysts from avian feces have become the basis for species identification. In contrast, molecular studies of wild birds' Coccidia are still in their infancy and are mostly based on DNA extracted from the blood stages of these parasites. Linking microscopic and molecular data requires a method that reliably extracts DNA from single oocysts with parallel detailed morphological examination of the same cell. We offer a thorough manual of isolating, photographing, and trapping single oocysts from avian feces, followed by extraction of parasite DNA and amplification of mitochondrial DNA from the same cells. In 39 single oocysts from 6 wild blackcaps, we combined microscopic studies of individual cells with studies on their mitochondrial haplotype. In 72% of the single oocysts sampled, we detected unambiguous sequences. From feces and blood of investigated birds, we obtained 6 different haplotypes of Isospora sp. (iSAT1-iSAT 6), finding both the same haplotype in different host individuals and various haplotypes in the same host individual. Our described methodology enables linking the huge amount of morphological data with innovative gene analysis. This method expands the scope of genetic studies conducted on Isospora species, including routine molecular analysis of single oocysts isolated from fecal samples.

UI	MeSH Term	Description	Entries
D007549	Isospora	A genus of protozoan parasites found in the intestines of birds, amphibians, reptiles, and mammals, including man. The oocysts produce two sporocysts, each with four sporozoites. Many species are parasitic in wild and domestic animals.	Isosporas
D010784	Photomicrography	Photography of objects viewed under a microscope using ordinary photographic methods.	Photomicrographies
D010802	Phylogeny	The relationships of groups of organisms as reflected by their genetic makeup.	Community Phylogenetics,Molecular Phylogenetics,Phylogenetic Analyses,Phylogenetic Analysis,Phylogenetic Clustering,Phylogenetic Comparative Analysis,Phylogenetic Comparative Methods,Phylogenetic Distance,Phylogenetic Generalized Least Squares,Phylogenetic Groups,Phylogenetic Incongruence,Phylogenetic Inference,Phylogenetic Networks,Phylogenetic Reconstruction,Phylogenetic Relatedness,Phylogenetic Relationships,Phylogenetic Signal,Phylogenetic Structure,Phylogenetic Tree,Phylogenetic Trees,Phylogenomics,Analyse, Phylogenetic,Analysis, Phylogenetic,Analysis, Phylogenetic Comparative,Clustering, Phylogenetic,Community Phylogenetic,Comparative Analysis, Phylogenetic,Comparative Method, Phylogenetic,Distance, Phylogenetic,Group, Phylogenetic,Incongruence, Phylogenetic,Inference, Phylogenetic,Method, Phylogenetic Comparative,Molecular Phylogenetic,Network, Phylogenetic,Phylogenetic Analyse,Phylogenetic Clusterings,Phylogenetic Comparative Analyses,Phylogenetic Comparative Method,Phylogenetic Distances,Phylogenetic Group,Phylogenetic Incongruences,Phylogenetic Inferences,Phylogenetic Network,Phylogenetic Reconstructions,Phylogenetic Relatednesses,Phylogenetic Relationship,Phylogenetic Signals,Phylogenetic Structures,Phylogenetic, Community,Phylogenetic, Molecular,Phylogenies,Phylogenomic,Reconstruction, Phylogenetic,Relatedness, Phylogenetic,Relationship, Phylogenetic,Signal, Phylogenetic,Structure, Phylogenetic,Tree, Phylogenetic
D005243	Feces	Excrement from the INTESTINES, containing unabsorbed solids, waste products, secretions, and BACTERIA of the DIGESTIVE SYSTEM.
D006239	Haplotypes	The genetic constitution of individuals with respect to one member of a pair of allelic genes, or sets of genes that are closely linked and tend to be inherited together such as those of the MAJOR HISTOCOMPATIBILITY COMPLEX.	Haplotype
D000818	Animals	Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA.	Animal,Metazoa,Animalia
D001715	Bird Diseases	Diseases of birds not considered poultry, therefore usually found in zoos, parks, and the wild. The concept is differentiated from POULTRY DISEASES which is for birds raised as a source of meat or eggs for human consumption, and usually found in barnyards, hatcheries, etc.	Avian Diseases,Avian Disease,Bird Disease,Disease, Avian,Disease, Bird,Diseases, Avian,Diseases, Bird
D012680	Sensitivity and Specificity	Binary classification measures to assess test results. Sensitivity or recall rate is the proportion of true positives. Specificity is the probability of correctly determining the absence of a condition. (From Last, Dictionary of Epidemiology, 2d ed)	Specificity,Sensitivity,Specificity and Sensitivity
D016054	DNA, Protozoan	Deoxyribonucleic acid that makes up the genetic material of protozoa.	Protozoan DNA
D016133	Polymerase Chain Reaction	In vitro method for producing large amounts of specific DNA or RNA fragments of defined length and sequence from small amounts of short oligonucleotide flanking sequences (primers). The essential steps include thermal denaturation of the double-stranded target molecules, annealing of the primers to their complementary sequences, and extension of the annealed primers by enzymatic synthesis with DNA polymerase. The reaction is efficient, specific, and extremely sensitive. Uses for the reaction include disease diagnosis, detection of difficult-to-isolate pathogens, mutation analysis, genetic testing, DNA sequencing, and analyzing evolutionary relationships.	Anchored PCR,Inverse PCR,Nested PCR,PCR,Anchored Polymerase Chain Reaction,Inverse Polymerase Chain Reaction,Nested Polymerase Chain Reaction,PCR, Anchored,PCR, Inverse,PCR, Nested,Polymerase Chain Reactions,Reaction, Polymerase Chain,Reactions, Polymerase Chain