Phylogenetic Relationships among Eight Eimeria Species Infecting Domestic Fowl Inferred Using Complete Small Subunit Ribosomal DNA Sequences

Abstract

Complete 18S ribosomal RNA gene sequences were determined for 8 Eimeria species of chickens and for Eimeria bovis of cattle. Sequences were aligned with each other and with sequences from 2 Sarcocystis spp., Toxoplasma gondii, Neospora caninum, and 4 Cryptosporidium spp. Aligned sequences were analyzed by maximum parsimony to infer evolutionary relationships among the avian Eimeria species. Eimecia bovis was found to be the sister taxon to the 8 Eimeria species infecting chickens. Within the avian Eimeria species, E. necatrix and E. tenella were sister taxa: this clade attached basally to the other chicken coccidia. The remaining Eimeria spp. formed 3 clades that correlated with similarities based on oocyst size and shape. Eimeria mitis and Eimeria mivati (small, near spherical oocysts) formed the next most basal clade followed by a clade comprising Eimeria praecox. Eimeria maxima, and Eimeria brumetti (large, oval oocysts), which was the sister group to Eimeria acervulina (small, oval oocysts). The 4 clades of avian Eimeria species were strongly supported in a bootstrap analysis. Basal rooting of E. necatrix and E. tenella between E. bovis and the remaining Eimeria species and the apparent absence of coccidia that infect the ceca of jungle fowl all suggest that E. necatrix and E. tenella may have arisen from a host switch, perhaps from the North American turkey, Meleagris gallopavo.

Supplementary studies on Henneguya doneci Schulman, 1962 (Myxozoa: Myxosporea) infecting the gill filaments of Carassius auratus gibelio (Bloch) in China: histologic, ultrastructural, and molecular data

“…Several cysts, collected from three infected fish, were ground using a pestle and the total DNA was extracted with SBS UltraPure™ Genomic DNA extraction kit (SBS Genetech Co., Ltd, Shanghai) according to the manufacturer's protocol. The gene was amplified on an MJ Research PTC-200 thermocycler with the primer pairs 18e (Hillis and Dixon 1991) and ERIB10 (Barta et al 1997). The polymerase chain reaction (PCR) was performed in 50 μL volumes consisting of approximately 1 μL of extracted DNA, 5 μL of 10× PCR buffer, 2.5 U of Ex Taq (Takara, Japan), 5 μL of 10× Ex Taq buffer, 0.25 mM dNTP mixture (Takara, Japan), and 40 pM of each primer.…”

Supplemental description and molecular characterization of Myxobolus miyarii Kudo, 1919 (Myxosporea: Myxobolidae) infecting intestine of Amur catfish (Silurus asotus)

“…The 18S small subunit ribosomal DNA (18S rDNA) gene was amplified by the nested-PCR, firstly using the ERIB1-ERIB10 primer pair (Barta et al 1997) in a 25 μl reaction mixture, containing 30 ng template DNA, 1 × PCR mixture (CWbiotech, China), 10 pmol of each primer. The PCR cycle consisted of an initial denaturation step at 95°C for 4 min, followed by 35 cycles at 95°C for 1 min, 48°C for 1 min, 72°C for 2 min, and a final extension at 72°C for 10 min.…”

Three actinosporean types (Myxozoa) from the oligochaete Branchiura sowerbyi in China

“…Genomic DNA was extracted using the QIAamp DNA Mini Kit (Qiagen Inc.). The partial 18S small-subunit ribosomal DNA gene was first amplified with the universal eukaryotic primers ERIB1 and ERIB10 (Barta et al 1997). And then, a nest PCR reaction was carried out employing more specific myxozoan primers MyxospecF and MyxospecR (Fiala 2006).…”