Ophryocystis sitonae sp. nov., (Neogregarinida: Ophryocystidae) parasitizing Sitona humeralis Stephens, 1831 (Coleoptera: Curculionidae).

Sitona humeralis Stephens 1831 (Coleoptera: Curculionidae) is an important pest of the Medicago and Vicia species in Turkey, and this study was conducted the determine the natural pathogens of this beetle. In the present study, a new neogregarine was observed in Malpighian tubules of the S. humeralis, collected from Ordu (Turkey) on the wild Medicago species. The yellowish oocysts were the most notable feature of the current neogregarine. The Giemsa-stained mature oocysts were fusiform shaped and measured 8.7 ± 0.7 (7.12-11.11) µm in length and 4.1 ± 0.3 (3.05-5.01) µm in width. The smooth oocyst wall was relatively thin (175-230 nm), and polar plugs were non-evident (weight = 380 nm, height = 500 nm). The 18S rDNA gene of the current neogregarine was sequenced and compared with fifteen sequences from GenBank. Morphological, ultrastructural, and molecular features indicate that the described neogregarine in S. humeralis differed from the all known Ophryocystis species and named here Ophryocystis sitonae sp. nov.

First record and distribution of Adelina sp. (Coccidia: Adeleidae) in populations of the Anisoplia segetum Herbst (Coleoptera: Scarabaeidae) in Turkey.

In this study, an Adelina sp. (Coccidia) infection was determined for the first time from Anisoplia segetum (Coleoptera: Scarabaeidae). The infection was observed in the gut and fat body of the host. The determined oocysts were ellipsoidal and measured 36.3 (34-38) × 30.2 (27-32) µm. The number of sporocysts per oocyst varied from 7 to 15. The dizoic sporocysts were usually subspherical, 10.2 (9-12) × 8.9 (8-10) µm. Also, some developmental stages and unsporulated oocysts were determined with electron microscopical observations. The immature microgamonts were observed in different shapes, from ovoid to spherical. And the detected oval unsporulated oocyst was measured 4.9 µm in length and 3.6 µm in width. The formation and distribution of these pathogens were also presented in A. segetum populations.

Assignment of Vairimorpha leptinotarsae comb. nov. on the basis of molecular characterization of Nosema leptinotarsae Lipa, 1968 (Microsporidia: Nosematidae).

Nosema leptinotarsae Lipa, 1968 is a microsporidian pathogen of the Colorado potato beetle, Leptinotarsa decemlineata Say. (Coleoptera: Chrysomelidae). To determine the phylogenetic status of N. leptinotarsae, the 16S SSU rRNA gene was sequenced (GenBank Accession No. MN841279) and compared phylogenetically against 21 microsporidian 16S SSU rRNA sequences using neighbour-joining and maximum-parsimony methods. The per cent identities of the N. leptinotarsae and other members of the Nosema-Vairimorpha clade ranged from 78.1 to 98.5%. Pairwise phylogenetic distances between the N. leptinotarsae and other species ranged from 0.009 to 0.320. Phylogenetic analysis shows clearly that N. leptinotarsae is a member of the Vairimorpha clade rather than the Nosema clade. The sequence divergence and morphological traits separated the N. leptinotarsae from other species in the Vairimorpha complex. As a result, a new assignment of Vairimorpha leptinotarsae comb. nov. has been implemented for N. leptinotarsae according to the phylogenetical positioning in the present study.

Ultrastructural and molecular characterization of Nosema alticae sp. nov. (Microsporidia: Nosematidae), pathogen of the flea beetle, Altica hampei Allard, 1867 (Coleoptera: Chrysomelidae).

In this study, the first microsporidian pathogen from Altica hampei (Coleoptera: Chrysomelidae) is described based on light microscopy, ultrastructural characteristics and comparative 16S SSU rDNA analysis. All developmental stages of the microsporidium are diplokaryotic and in direct contact with the host cell cytoplasm. Giemsa-stained mature spores are oval in shape and measured 3.82 ± 0.35 µm in length and 2.54 ± 0.27 µm in width. The polar filament of the binucleate spores is isofilar with 12-14 coils. Coils are 140.28 ± 4.88 nm (135.59-147.06; n = 36) in diameter and consist of six concentric layers of different electron density and thickness. The spores have a relatively thick (161.72 ± 29.19 nm) trilaminar spore wall. Morphological, ultrastructural and molecular features indicate that the described microsporidium belongs to the genus Nosema and is named Nosema alticae sp. nov.

A new microsporidium, Vairimorpha subcoccinellae n. sp. (Microsporidia: Burenellidae), isolated from Subcoccinella vigintiquatuorpunctata L. (Coleoptera: Coccinellidae).

A new microsporidium was isolated from Subcoccinella vigintiquatuorpunctata L. (Coleoptera: Coccinellidae), a pest of Galega officinalis L. in Turkey. Infection in larval and adult stages was systemic with mature spores produced in the midgut, gonads, Malpighian tubules and, most extensively, fat body tissues. The microsporidium was polymorphic with two sporulation sequences producing two types of spores, binucleate spores with 13-15 coils of the polar tube, and uninucleate spores with 7 coils of the polar tube that developed within a sporophorous vesicle (SPV) to form meiospores. The 16S small subunit rRNA (SSU rRNA) gene of the microsporidium was sequenced and compared with twenty-seven microsporidian sequences from GenBank. Based on the phylogenetic analysis of the SSU rRNA sequence, this microsporidium is unique within the Vairimorpha group. Morphological and genetic characters indicate that the described microsporidium is dissimilar to all known Vairimorpha species, and so is named here as Vairimorpha subcoccinellae n. sp.

Aranciocystis muskarensis n. gen., n. sp., a neogregarine pathogen of the Anisoplia segetum Herbst (Coleoptera: Scarabaeidae).

In this study, a new genus and species of neogregarine which is a pathogen of Anisoplia segetum Herbst (Coleoptera: Scarabaeidae), is described. The adult beetles of A. segetum were collected from Nevsehir, Turkey and neogregarine infection rates were determined as 18.52%. The Giemsa-stained mature oocysts are lemon-shaped and measured 9.34±0.82µm in length and 5.77±0.77µm in width. The oocyst wall surface of the mature oocysts is similar to an osage orange (tuberculate). Morphological, ultrastructural and molecular features indicate that the previously undescribed neogregarine is dissimilar to all known neogregarine taxa and represents the first record from Anisoplia segetum and is named here as Aranciocystis muskarensis n. gen., n. sp.

Description and phylogeny of a new microsporidium from the elm leaf beetle, Xanthogaleruca luteola Muller, 1766 (Coleoptera: Chrysomelidae).

This study describes a new genus and species of microsporidia which is a pathogen of the elm leaf beetle, Xanthogaleruca luteola Muller, 1776 (Coleoptera: Chrysomelidae). The beetles were collected from Istanbul in Turkey. All developmental stages are uninucleate and in direct contact with the host cell cytoplasm. Giemsa-stained mature spores are oval in shape and measured 3.40 ± 0.37 µm in length and 1.63 ± 0.20 µm in width. These uninucleate spores have an isofilar polar filament with 11 turns. The spore wall was trilaminar (75 to 115 nm) with a rugose, electron-dense exospore (34 to 45 nm) and a thickened, electron-lucent endospore (65 to 80 nm) overlaying the plasmalemma. Morphological, ultrastructural, and molecular features indicate that the described microsporidium is dissimilar to all known microsporidian taxa and confirm that it has different taxonomic characters than other microsporidia infecting X. luteola and is named here as Rugispora istanbulensis n. gen., n. sp.

The first record of nucleopolyhedrovirus isolated from the gypsy moth, Lymantria dispar (Lepidoptera, Lymantriidae) in Turkey.

OBJECTIVE: The gypsy moth, Lymantria dispar L. (Lepidoptera, Lymantriidae) is a common pest of forests and fruit trees throughout the world. This insect is also a major serious pest in Turkey. Nowadays L. dispar can be managed by biological control methods especially, using entomopathogenic viruses. The aim of this study is to characterize entomopathogenic viruses and is the first record of nucleopolyhedrovirus isolated from the gypsy moth in Turkey. METHODS: PIBs obtained from infected larvae were measured and photographed using an Olympus BX51 microscope with a DP-25 digital camera and a DP2-BSW Soft Imaging System and examined with a Philips 208 electron microscope (TEM). RESULTS: The virus had the typical characteristics of nucleopolyhedroviruses. The dimension of the polyhedral inclusion bodies (PIBs) was 2.03±0.25 µm. PIBs varied in size from 1.65 to 2.21 µm and were usually polygonal in shape. Virions in PIBs contained 1 to 8 nucleocapsids per virion. The size of the viral particles was 366.67±54.72 (312-500) x 42.95±6.12 (30-47) nm. CONCLUSION: The isolation and characterization of a pure isolate of Lymantria dispar multinucleopolyhedrovirus (LdMNPV-TR) from Turkey is presented for the first time.