Molecular and Morphological Characterization of Anncaliia azovica sp. n. (Microsporidia) Infecting Niphargogammarus intermedius (Crustacea, Amphipoda) from the Azov Sea

Abstract

Five out of one hundred adults of Niphargogammarus intermedius caught at the Azov sea shore were found to be infected with microsporidia. The infection was found in the subcuticular fat body and myocytes. Parasites developed in direct contact with host cells, displayed a disporoblastic sporogony and diplokaryotic arrangement of nuclei at all stages. Spores were oval, 4.6-5.8 × 2.6-3.0 μm. Exospore appendages, vesicular-tubular secretions, and the anisofilar polar filament indicated a similarity to Anncaliia species. Sporont surfaces displayed ridges of amorphous material. Meronts and sporonts formed protoplastic extensions, similar to A. vesicularum and A. meligheti. Mature spores possessed a bipartite polaroplast. The polar tube was arranged in one row of 13-18 coils including 0-3 distal coils of lesser diameter. Partial sequencing of SSU, ITS, and LSU regions of rRNA gene (GenBank accessions: KY288064-KY288065) confirmed this new species to be congeneric with A. algerae (#AF069063) and A. meligheti (#AY894423). The SSU gene of this novel microsporidium shared 99.4% sequence similarity to A. algerae and 98.9% to A. meligheti. Genes for HSP70 and RPB1 amplified with primers designed for A. algerae orthologs displayed 99.7% and 97.4% similarity, respectively, between A. algerae and the novel microsporidium. A new species, Anncaliia azovica, is described based on morphological and molecular characterization.

Development of Anncaliia algerae (Microsporidia) in Drosophila melanogaster

“…The protoplasmic extensions were first described by Cali et al (1998) as a characteristic feature of Anncaliia (Brachiola) vesicularis. They were later observed in A. azovica (Tokarev et al 2018). Our current study proves that A. algerae stages also exhibit these processors, which are, likely, common to all (or at least several) Anncaliia spp., but can be easily overseen during EM analysis.…”

Development of Microsporidiology in Russia

“…Использование молекулярно-филогенетического анализа сделало возможным выяснение родственных отношений видов, относящихся к разным родам, в нескольких группировках микроспоридий. Подтверждена валидность ранее описанных видов микроспоридий пресноводных ракообразных (Issi et al, 2012;Tokarev et al, 2012) и рыб (Токарев и др., 2015), прямокрылых (Issi et al, 2008), жесткокрылых (Franzen et al, 2006) и чешуекрылых насекомых , составлены полные описания новых видов (Issi et al, 2010;Malysh et al, 2013;Simakova et al, 2018;Tokarev et al, 2018b), в том числе типовых видов новых родов (Tokarev et al, 2010ab;Lipa et al, 2020), проведена дифференциальная диагностика микроспоридий, обнаруженных в лабораторных и производственных культурах насекомых (Tokarev et al, 2018a), в том числе двух микроспоридий медоносной пчелы в условиях Западной Сибири (Tokarev et al, 2018c). Очень интересна и своевременна работа по ревизии всего состава видов, отнесённых к двум родам -Nosema и Vairimorpha, для уточнения диагнозов этих родов с учётом всех последних молекулярно-филогенетических данных (Tokarev et al, 2020).…”

Intraspecific genetic diversity of the fish-infecting microsporidian parasite Pseudokabatana alburnus (Microsporidia)

“…The Rpb1 gene was also applied into the species classification and intraspecific variation of microsporidia ( Gomez-Moracho et al, 2014 ; Maside et al, 2015 ; Kyei-Poku and Sokolova, 2017 ; Pretto et al, 2018 ; Tokarev et al, 2018 , 2020 ; Yurakhno et al, 2021 ), which was generally thought to be of single copy and possesses more variable sites than those of the SSU rDNA, However, the amplification of Rpb1 is difficult and only a small amount of Rpb1 gene sequence is so far available in the public database. In the present study, failing to amplify Rpbl of some P. alburnus isolates with the same primer pairs and conditions can partially explain the problem.…”