Wings’ Descriptive and Geometric Morphometry Analyses Support Distinct Phlebotomus sergenti Parrot Populations in Central Morocco

Phlebotomus (Paraphlebotomus) sergenti Parrot, 1917, the proven vector of Leishmania tropica Wright, 1903, the causative agent of anthroponotic cutaneous leishmaniasis, is widely distributed in Morocco. Previous works using molecular markers (ITS2 rDNA and Cyt b mtDNA) have hy-pothesized the existence of multiple closely related populations of sandfly species (cryptic species) that would exhibit distinct vectorial capacities. In this work, descriptive characteristics of wings (size and shape of the right and left wings) were measured in samples collected from fourteen sta-tions in central Morocco. These analyses support the existence of distinct P. sergenti populations, enlightening significant phenotypic variations of P. sergenti’s wings, regarding their size and shape, depending on geographic origin. In addition, geomorphometric analyses of wing’s length, centroid size, alpha, and beta distances allowed clear discrimination of P. sergenti sub-populations. These data pinpoint the adaptative ability of P. sergenti to local environmental conditions. Additional studies are now required to further shed light on the genetic structure of P. sergenti populations in Morocco.

The Structural Difference of Isobaric N-Glycans of Two Microalgae Samples Reveals Taxonomic Distance

Microalgae of the Chlorella clade are extensively investigated as an environmentally friendly source of renewable biofuels and high-value nutrients. In addition, essentially unprocessed Chlorella serves as wholesome food additive. A recent study on 80 commercial Chlorella preparations revealed an unexpected variety of protein-linked N-glycan patterns with unprecedented structural features, such as the occurrence of arabinose. Two groups of products exhibited a characteristic major N-glycan isobaric to the Man2GlcNAc2XylFuc N-glycan known from pineapple stem bromelain, but tandem mass spectrometry (MS/MS) analysis pointed at two types of N-glycan different from the bromelain structure, as well as from each other. Here we report the exact structures of these two novel N-glycan structures, elucidated by nuclear magnetic resonance spectroscopy and MS/MS, as well as on their phylogenetic context. Despite their humble size, these two N-glycans exhibited a very different design with structural features unrelated to those recently described for other Chlorella-clade strains. The major glycans of this study presented several novel structural features such as substitution by arabinose or xylose of the internal N-acetylglucosamine, as well as methylated sugars. ITS1-5.8S-ITS2 rDNA barcode analyses revealed that the xylose-containing structure derived from a product primarily comprising Scenedesmus species, and the arabinose-containing glycan type related to Chlorella species (SAG211-34 and FACHB-31) and to Auxenochlorella. This is another example where characteristic N-glycan structures distinguish phylogenetically different groups of microalgae.

Gerronema nemorale: first report of the genus and species from Pakistan

Basidiomata of a Gerronema species, collected in Murree, Punjab, Pakistan, have been confirmed as G. nemorale by morphological and ITS1-5.8S-ITS2 rDNA sequence analyses. This is the first report of the genus Gerronema from Pakistan, and the first record of G. nemorale for the South Asian region.

Morphology and molecular data of the species of Suillus (Suillaceae, Boletales) associated with Pinus sibirica at the European northeast of Russia

The genus Suillus represents the largest genus of the family Suillaceae, forming ectomycorrhizae with coniferous trees. Here, we presented the first results of molecular research (nrITS1-5.8S-ITS2 rDNA sequences) of the specimens of Suillus, forming ectomycorrhizae with Pinus sibirica, at the territory of European northeast of Russia. The six taxa (Suillus acidus var. intermedius, S. placidus, S. plorans subsp. plorans, S. plorans subsp. cyanescens, S. punctipes and S. sibiricus) are identified based on morphological and molecular evidence. The key for identification, full morphological descriptions, the color photos of basidiomata and information on their ecology and distribution in Russia and world are presented for these taxa. On the basis of the molecular approach, we confirmed the relevance of subspecies Suillus plorans subsp. cyanescens previously described only by morphological features. BLAST-analysis of Suillus placidus with untypical for this species brown pileus revealed 99% identity with the specimens of S. placidus from Asia, for which the color of pileus varies from white to brown.

Changes in the Splenic Melanomacrophage Centre Surface Area in Southern Bluefin Tuna (Thunnus maccoyii) Are Associated with Blood Fluke Infections

Melanomacrophage centres (MMCs) are aggregates of macrophages accumulating various pigments. They have been proposed as an indicator of fish immune response. Blood flukes are common parasites in farmed fish. Two cohorts of wild Southern Bluefin Tuna (Thunnus maccoyi) were examined at transfer, before treatment against blood flukes (pre-treatment) and at harvest. MMCs were assessed in histological sections using image analysis, while Cardicola forsteri and Cardicola orientalis infection severity was determined using qPCR, count of adult flukes in heart flushes and count of eggs in gill filaments. Fish from both cohorts showed the same pattern in the changes in the surface area of MMCs. The surface area of splenic MMCs increased over the ranching duration and was positively correlated to the PCR determined copy numbers of Cardicola forsteri ITS2 rDNA in the gills of those fish. However, the infection with blood fluke was more variable, both between cohorts and individuals within the same cohort. Eggs of blood fluke were detected in renal MMCs using histology. Cardicola forsteri had a higher prevalence than Cardicola orientalis. This study contributes to our understanding of blood fluke infections in Southern Bluefin Tuna and their interactions with MMCs.

Morphological and Molecular Evidence for the Occurrence of Hodophilus variabilipes (Clavariaceae, Agaricales) in Poland

<em>Hodophilus variabilipes</em>, a species hitherto unknown in Poland, has been reported from a single locality in the Pieniny Mts (S Poland). The first studies on <em>H. variabilipes </em>were those of macro- and microscopic features and molecular characteristics (ITS1-5.8S-ITS2 rDNA sequences). Here, a brief description and illustration of <em>H. variabilipes </em>based on Polish material are given, and the ecology and characteristics distinguishing <em>H. variabilipes </em>from related species of the genus <em>Hodophilus </em>are briefly discussed.

The first record of the trematode Urogonimus certhiae (Trematoda: Leucochloridiidae) in the Eurasian nuthatch Sitta europaea

Adult trematodes of the genus Urogonimus Monticelli, 1888 were found in the cloaca of a male Eurasian nuthatch Sitta europaea Linnaeus, 1758, the carcass of which was found in the Vyritsa Settlm. (Leningrad Prov., Northwest Russia). Based on morphological characters, the worms were identified as U. certhiae McIntosh, 1927. This is the first record of this parasite from the nuthatch and from Northwest Russia. We analysed nucleotide sequences of ITS1-5.8S-ITS2 rDNA of this species and found important differences with U. macrostomus (Rudolphi, 1803). Genetic and morphological data indicated that U. certhiae and U. macrostomus were two separate species. Phylogenetic analysis confirmed that Urogonimus and Leucochloridium Carus, 1835 were two distinct genera.

Sequence analysis of the internal transcribed spacer 2 (ITS2) region of rDNA for identifying Trichogramma species and evaluating genetic diversity

Species of Trichogramma Westwood, 1833 (Hymenoptera: Trichogrammtidae) are frequently used as biological control agents against Lepidoptera, but practical application of these egg endoparasitoids are complicated because of their complex taxonomy. This study aimed to compare sequences of internal transcribed spacer regions of ribosomal DNA (ITS2-rDNA) of Trichogramma accessions with those deposited in GenBank in order to access the reliability of the ITS2 as a barcode for discriminating species and evaluating the genetic diversity. ITS2-rDNA sequences obtained from seventeen specimens of Trichogramma confirmed previous identifications based on morphological characteristics. Multiple sequence alignment revealed the existence of highly conserved regions in ITS2 sequences while the neighbour-joining dendrogram indicated that the specimens formed three clusters comprising T. manicobai and T. marandobai (group I), T. galloi (group II) and T. pretiosum (group III). The ITS2 marker was shown to be a powerful DNA barcode for discriminating Trichogramma species and could be used to complement the morphological approach.

Comparative Phylogeography and Integrative Taxonomy of Ochlerotatus caspius (Dipera: Culicidae) and Ochlerotatus dorsalis

Given that accurately identifying pathogen vectors is vital for designing efficient mosquito control programs based on the proper surveillance of the epidemiologically important species, it has been suggested the complementary use of independently evolving genes and morphometric traits as a reliable approach for the characterization and delimitation of related species. Hence, we examined the spatial distribution of COI mtDNA and ITS2 rDNA variation from the historical perspective of Ochlerotatus caspius (Pallas, 1771) and O. dorsalis (Meigen, 1830), while simultaneously testing the utility of the two markers in integrative species delimitation when combined with phenotypic character analyses of larvae and adults. Despite the striking difference in haplotype diversity (high in COI mtDNA, low in ITS2 rDNA), no evident phylogeographic structure was apparent in the Palearctic O. caspius. The Holarctic O. dorsalis species was subdivided into two highly distinctive COI mtDNA phylogroups which corresponded to the Nearctic and Palearctic regions. Strong support for the independence of the two allopatric evolutionary lineages suggested that geographical barrier and climatic changes during Pleistocene caused vicariance of the ancestral range. COI mtDNA reliably distinguished O. caspius and O. dorsalis, while ITS2 rDNA yet again lacked the proper resolution for solving this problem. An integrative approach based on the larval and adult morphological traits have varying taxonomic applications due to their differential diagnostic values. Thus, by the implementation of an integrative taxonomic approach, we successfully detected species borders between the two epidemiologically relevant species and uncovered the presence of cryptic diversity within O. dorsalis.

Caterpillar gut and host plant phylloplane mycobiomes differ: a new perspective on fungal involvement in insect guts

ABSTRACT Compared with the highly diverse microbiota of leaves, herbivorous insects exhibit impoverished gut microbial communities. Research to date has focused on the bacterial component of these gut microbiomes, neglecting the fungal component. As caterpillar gut bacterial microbiomes are derived strongly from their diet, we hypothesized that their mycobiomes would reflect the host leaf mycobiomes. Using the ITS2 rDNA and V5–V6 16S rRNA gene regions for DNA metabarcoding of caterpillar gut and host leaf sample pairs we compared their mycobiome genus diversity and compositions and identified genera associated with caterpillar guts. Leaves and caterpillar guts harbored different mycobiomes with quite low qualitative similarity (Jaccard index = 38.03%). The fungal genera most significantly associated with the caterpillar gut included Penicillium, Mucor and unidentified Saccharomycetales, whereas leaf-associated genera included Holtermanniella, Gibberella (teleomorph of Fusarium) and Seimatosporium. Although caterpillar gut and leaf mycobiomes had similar genus richness overall, this indicator was not correlated for individual duplets. Moreover, as more samples entered the analysis, mycobiome richness increased more rapidly in caterpillar guts than in leaves. The results suggest that the mycobiota of the caterpillar gut differs from that of their feeding substrate; further, the mycobiomes appear to be richer than the well-studied bacterial microbiotas.