During the investigation of parasitic pathogens of Mytilus coruscus, infection of a Perkinsus-like protozoan parasite was detected by alternative Ray's Fluid Thioglycolate Medium (ARFTM). The diameter of hypnospores or prezoosporangia was 8–27 (15.6 ± 4.0, n = 111) μm. The prevalence of the Perkinsus-like species in M. coruscus was 25 and 12.5% using ARFTM and PCR, respectively. The ITS1-5.8S-ITS2 fragments amplified by PCR assay had 100% homology to that of P. beihaiensis, suggesting that the protozoan parasite was P. beihaisensis and M. coruscus was its new host in East China Sea (ECS). Histological analysis showed the presence of trophozoites of P. beihaiensis in gill, mantle and visceral mass, and the schizonts only found in visceral mass. Perkinsus beihaiensis infection led to inflammatory reaction of hemocyte and the destruction of digestive tubules in visceral mass, which had negative effect on health of the farmed M. coruscus and it deserves more attention.

The Conyza genus includes nearly 150 species, comprising closely related weedy species. Proper identification of Conyza spp. is essential to develop effective strategies for their management. The overlap of traits, species varieties, and the putative occurrence of hybridization hampers the identification of Conyza spp. and its management in agricultural and natural environments. Herein, we assessed five DNA barcodes and 32 morphological traits to classify Conyza spp. and survey their dispersion in soybean fields [Glycine max (L.) Merr.] in Brazil in 2019, 2020, and 2021. The Conyza accessions included two species, hairy fleabane [Conyza bonariensis (L.) Cronquist) and Sumatran fleabane [Conyza sumatrensis (Retz.) E. Walker], and each species comprised two varieties. The ITS and rps16-trnQ gene regions showed the ability to distinguish between the two Conyza species, while the matK, rbcL, and trnF-trnF gene regions were not polymorphic. Out of 32 morphological traits, phyllary color, involucre shape, capitulescence type, and inflorescence type were the most polymorphic and even reliable for taxonomic purposes. The combination of ITS or ITS+rps16-trnQ regions and the four morphological markers was able to discriminate 91% of the plants, except those of C. bonariensis var. angustifolia. These results support the taxonomic resolution between C. bonariensis and C. sumatrensis and are useful for other Conyza spp. and other closely related weedy species worldwide. Conyza sumatrensis was detected in 94% of soybean fields across macroregions and seasons in Brazil, while C. bonariensis was sparsely dispersed, mainly in the southern macroregion (MRS 1).

Four species of Porpidia are newly reported from China, including one species new to science (Porpidia crystallina) and three records (Porpidia umbonifera, P. seakensis and P. cf. contraponenda) new to China. Porpidia crystallina is characterized by a macrocarpa-type exciple containing crystals, a Cinereorufa-green epihymenium, large ascospores and a lack of secondary metabolites. Morpho-anatomical, chemical and phylogenetic analyses were carried out to elucidate the placement of the species and to support the delimitation of the new taxon. Detailed taxonomic descriptions, ecological and chemical characters, and illustrations are provided for each species. A key to all known Chinese Porpidia species is also provided.

Infection by parasites or pathogens can have marked physiological impacts on individuals. In birds, infection may affect moult and feather growth, which is an energetically demanding time in the annual cycle. Previous work has suggested a potential link between clinically visible Trichomonas gallinae infection and wing length in turtle doves Streptopelia turtur arriving on breeding grounds. First, T. gallinae infection was characterized in 149 columbids from 5 species, sampled on turtle dove wintering grounds in Senegal during the moulting period, testing whether infection by T. gallinae is linked to moult. Trichomonas gallinae prevalence was 100%, so rather than testing for differences between infected and uninfected birds, we tested for differences in moult progression between birds infected by different T. gallinae strains. Twelve strains of T. gallinae were characterized at the internal transcribed spacer 1 (ITS1)/5.8S/ITS2 region, of which 6 were newly identified within this study. In turtle doves only, evidence for differences in wing length by strain was found, with birds infected by strain Tcl-1 having wings nearly 6 mm longer than those infected with strain GEO. No evidence was found for an effect of strain identity within species on moult progression, but comparisons between infected and uninfected birds should be further investigated in species where prevalence is lower.

This paper describes Diegloglossidium maradonai n. g., n. sp. a parasite of the intestine of Hoplosternum littorale (Hancock) from La Plata River basin. The new genus is morphologically similar to members of Alloglossidiidae and Macroderoidiidae although they also share some traits observed in both families. Those families can be differentiated from each other by the combination of morphological features, including the density and distribution of the tegumental spines, the distribution of the vitelline follicles and the extent of the post-testicular space. The molecular analyses based on the large subunit of the ribosomal RNA gene, and the internal transcribed spacer (ITS) regions including ITS1, 5.8S and ITS2 unequivocally place the new genus in the family Alloglossidiidae which is amended based on new observed features. Diegoglossidium n. g. is characterized by a combination of characteristics, being most notably the presence of a deeply lobed ovary. Lastly, the geographical distribution and host associations of the two closely related Neotropical genera of Alloglossidiidae: Magnivitellinum and Diegoglossidium are discussed, and the host and distribution range of Magnivitellinum saltaensis is expanded into Argentina.

Since 2017, a sea anemone species has been registered in central Portugal with a morphology comparable to Phymactis papillosa, from the Pacific shore of south and central America. In this paper, we have collected two individuals from Cascais, Portugal, extracted their DNA and amplified and sequenced the internal transcribed spacers I and II, markers frequently used in Cnidaria barcoding. The sequences obtained were compared with the ones available on GenBank, using the Basic Local Alignment Search Tool (BLAST) on the NCBI website. This procedure allowed the identification of P. papillosa. This is the first confirmed record of P. papillosa in European coasts. Citizen science data suggest occurrences in the Alentejo coast (south-west Portugal) and Mediterranean coast around Murcia, although no genetic or morphological confirmations of these records have been made. The potential effects of this species introduction are discussed.

The widespread species Parmotrema crinitum (Ach.) M. Choisy and Parmotrema perlatum (Huds.) M. Choisy are mainly distinguished by their reproductive strategies. While P. crinitum propagates by isidia, P. perlatum produces soredia. In this study, we aim to evaluate the phylogenetic relationship between both species and to critically examine their species boundaries. To this purpose, 46 samples belonging to P. crinitum and P. perlatum were used in our analysis, including 22 for which we studied the morphology and chemistry, before extracting their DNA. We used 35 sequences of the internal transcribed spacer region of nuclear ribosomal DNA (ITS) of Parmotrema perlatum from Europe and Africa (20 of which were newly generated), and 11 of Parmotrema crinitum from Europe, North America and North Africa (two newly generated). Additionally, 28 sequences of several species from Parmotrema were included in the ITS dataset. The ITS data matrix was analyzed using different approaches, such as traditional phylogeny (maximum likelihood and Bayesian analyses), genetic distances, automatic barcode gap discovery (ABGD) and the coalescent-based method poisson tree processes (PTP), in order to test congruence among results. Our results indicate that all samples referred to P. crinitum and P. perlatum nested in a well-supported monophyletic clade, but phylogenetic relationships among them remain unresolved. Delimitations inferred from PTP, ABGD and genetic distance analyses were comparable and suggested that P. crinitum and P. perlatum belong to the same lineage. Interestingly, two samples of P. perlatum separate in a different monophyletic clade, which is supported as a different lineage by all the analyses.

Twenty-eight species of Verrucariaceae are reported from Nepal. One genus and nine species are described as new: Nesothele gen. nov., sister to Staurothele s. lat., with a crustose to squamulose thallus, hymenial algae and 4–8 colourless muriform ascospores per ascus; Nesothele glebulosa sp. nov., resembling N. rugulosa but with smaller perithecia; Thelidium uvidulum sp. nov., producing a thin thallus with soralia, prominent perithecia, and 1-septate ascospores; Verrucaria antepotens sp. nov., having a well-developed thallus with dark-sided areoles, immersed perithecia, and small ascospores 12.5–16.5 μm long; V. lactea sp. nov., resembling V. praetermissa but with larger ascospores and a strongly deviating ITS sequence; V. parvipeltata sp. nov., with brown, basally constricted areoles on an extensive dark prothallus; V. senta sp. nov., with a brown cracked thallus and prominent naked perithecia; Willeya eminens sp. nov., with prominent perithecia, and differing from W. protrudens in its ITS sequence; W. irrigata sp. nov. with a thallus cracked into dark-sided areoles, and relatively large ascospores 28.5–40 μm long; W. nepalensis sp. nov. with a cracked thallus, immersed perithecia and a distinctive ITS sequence. Three species complexes might contain new taxa, but wider geographical sampling is necessary before delimiting species: Thelidium minutulum, Verrucaria elaeomelaena and V. hydrophila. Five new combinations are made: Willeya honghensis comb. nov. (for Staurothele honghensis), Nesothele globosa comb. nov. (for Endocarpon globosum), N. hymenogonia comb. nov. (for Staurothele hymenogonia), N. rugulosa comb. nov. (for Staurothele rugulosa) and N. succedens comb. nov. (for Staurothele succedens). Four species are unidentified.

Trematode taxonomy is mainly based on the morphological traits of adults. The identification of metacercariae is challenging because such traits are not developed in larval forms, and they even may show some level of morphological variability. Studies testing the potential correspondence between morphological differences and genetic variation of parasites are still lacking. The metacercariae of Posthodiplostomum minimum are probably the diplostomids more widely distributed in North and Middle American freshwater fish, and their intraspecific morphological variability has been attributed to the effect exerted by the host. Here, we tested the hypothesis whether they represent a single species, or a species complex by assessing the genetic divergence and phylogenetic relationships of metacercariae sampled from several host species in a wide geographical range across Middle America. The internal transcribed spacers (ITS1-5.8S-ITS2), and the mitochondrial COI gene were sequenced for 124 and 55 metacercariae, respectively. Phylogenetic analysis inferred from ITS sequences uncovered six well-supported monophyletic lineages. The six lineages show no correspondence to any Posthodiplostomum species for which sequences are available thus far in GenBank. Lineages exhibit some degree of host specificity; Lineages I, II, IV and V are primarily parasites of cyprinodontiforms of the families Poeciliidae, Goodeidae, Profundulidae and Fundulidae. In poeciliids there are at least four candidate species of Posthodiplostomum, some of them occurring in sympatry; instead, Lineages II and VI are exclusively parasites of cichlids. This study contributes to our understanding of the diversity of larval forms of diplostomids and provides an opportunity to further study their life cycles.

The tropical areas of eastern Asia contain a high diversity of foliicolous lichens, including various species of the genus Badimia. Badimia xanthocampylidia W. C. Wang & J. C. Wei is described from tropical rainforests in southern China and Thailand based on morphology, chemistry, and combined mtSSU, ITS and nrLSU sequences. It is characterized by a pale green thallus with yellow verrucae and bright yellow campylidia and the presence of isousnic acid. Three other species, B. multiseptata Papong & Lücking, B. pallidula (Kremp.) Vězda and B. polillensis (Vain.) Vězda, are discussed and the genus Badimia is newly reported from China. A worldwide key to currently known species in the genus is presented.

In vitro activation of primordial follicles could serve as a safe method to preserve fertility in patients with cancer subjected to ovarian tissue cryopreservation during oncotherapy, however the culture medium for this purpose requires to be optimized. Granulosa cell conditioned medium (GCCM) has been recognized to enhance primordial follicle activation and the present study was conducted to understand whether addition of pyruvate, a combination of insulin, transferrin and selenium (ITS) or testosterone to GCCM could improve its efficiency in this regard. To this end, 1-day-old mouse ovaries were cultured in four different media including CON (control; containing GGCM only), PYR (containing GCCM plus pyruvate), ITS (containing GCCM plus ITS) or TES (containing GCCM plus testosterone) for 11 days. Furthermore, follicular dynamics and gene expression of factors involved in follicular development were assessed using histological examination and RT-PCR, respectively, on days 5 and 11 of culture. Pyruvate decreased follicular activation, but it enhanced the progression of follicles to the primary stage. Moreover, it upregulated Bmp15 and Cx37 (P < 0.05). In the ITS group, activation of follicles was not affected and total number of follicles was reduced by day 11 of culture. Additionally, ITS downregulated Pi3k, Gdf9, Bmp15 and Cx37 (P < 0.05). Although testosterone did not affect primordial follicle activation, it enhanced the development of follicles up to the preantral stage (P < 0.05). Furthermore, testosterone inhibited the expression of Pten but stimulated the expression of Gdf9 and Cx37 (P < 0.05). In conclusion, the present study revealed that inclusion of pyruvate and testosterone into GCCM could enhance the early development of follicles in cultured 1-day-old mouse ovaries.

Twelve hyper-β carotene-producing strains of algae assigned to the genus Dunaliella salina have been isolated from various hypersaline environments in Israel, South Africa, Namibia and Spain. Intron-sizing of the SSU rDNA and phylogenetic analysis of these isolates were undertaken using four commonly employed markers for genotyping, LSU rDNA, ITS, rbcL and tufA and their application to the study of Dunaliella evaluated. Novel isolates have been identified and phylogenetic analyses have shown the need for clarification on the taxonomy of Dunaliella salina. We propose the division of D. salina into four sub-clades as defined by a robust phylogeny based on the concatenation of four genes. This study further demonstrates the considerable genetic diversity within D. salina and the potential of genetic analyses for aiding in the selection of prospective economically important strains.

Lecanora s. lat. is a genus of crustose, rarely placodioid lichens comprising c. 1000 recognized species and subdivided into several morphology-based groups. Some of these groups have been supported in phylogenetic analyses and segregated as new genera. One of the remaining groups that has not been previously studied by molecular methods in much detail, the L. saligna-group, includes corticolous and lignicolous crustose lichens, usually containing isousnic or usnic acid (or both) as major secondary metabolites. As part of our ongoing project ‘Lecanomics’, a phylogenetic analysis based on two loci was conducted and found the L. saligna-group to be divided into two main clades and several well-supported minor clades. The L. varia clade, chosen as one of the outgroups, emerged within the L. saligna-group. The majority of the clades are characterized by phenotypic differences. However, several well-supported clades share similarities with their sister groups, suggesting that species circumscriptions based solely on phenotypic characters may be too conservative to characterize the true species diversity present within the group. Also, there is evidence for some geographical separation of lineages; for example, most North American individuals, previously known as Lecanora saligna and L. albellula, form two clades separate from their European namesakes and are here preliminarily called ‘Lecanora sp. B’, ‘Lecanora sp. C’ and ‘Lecanora sp. D’. However, L. saligna and L. albellula also appear to occur in North America, and some specimens from the Caucasus and Iran cluster within the North American clades. Lecanora anopta and L. subravida are reported for the first time from Iran.

The phylogeny of foliicolous taxa in Pilocarpaceae was reconstructed using Bayesian and maximum likelihood analyses of concatenated ITS and mtSSU sequences. Sixty-six new partial sequences representing 36 taxa were generated and 29 sequences were downloaded from GenBank. Our results indicate that Lasioloma R. Sant. is nested within a paraphyletic Calopadia Vězda, whereas Fellhanera Vězda is polyphyletic and the phylogenetic relationships of Eugeniella Lücking et al. and Sporopodium Mont. with Fellhanera require further investigation. In addition, Tapellaria parvimuriformis W. C. Wang & J. C. Wei is described as a new species from Khao Yai National Park, Thailand, and is characterized by its small muriform ascospores, grey campylidia with a pale base, and short conidia. Eight new records of foliicolous lichens for Thailand are also listed.

Porina collina Orange, Palice & Klepsland is described as new from siliceous rocks in the Czech Republic, Great Britain and Norway. The thallus produces isidioid propagules which resemble those in P. rosei, P. hibernica and P. pseudohibernica, but which are more fragile and poorly-defined, and almost ecorticate; the perithecial wall is dark purplish brown and the ascospores are 3-septate. Sequences of the mitochondrial ribosomal DNA suggest that the new species belongs in the Porina byssophila clade.

A new species, Huriella flakusii, is described from the arid region of the Colca Canyon in southern Peru, and is characterized by squamulose, orange thalli and abundant, crowded and aggregated, usually flexuous when compressed, reddish apothecia. The thalline cortex, proper margin and hypothecium are paraplectenchymatous. The ascospores are broadly ellipsoid, 10‒15 × 5.0‒9.5 µm, with rather thin septa, 2‒4 µm. Phylogenetic analyses of the DNA sequences of ITS, nuLSU and mrSSU rDNA markers revealed that the new species is nested within the Amundsenia-Squamulea subclade in the subfamily Xanthorioideae, and has a sister group relationship to Huriella loekoesiana from South Korea.

As part of our ongoing research on Peltigera, we recognize a morphologically and phylogenetically distinct new species, Peltigera shennongjiana L. F. Han & S. Y. Guo, from the Shennongjia region of Central China. It is distinguished from other members of the P. canina-group by the presence of abundant phyllidia and flat, branched lobules along the margin or laminal cracks, short lobes, and a pruinose, usually greyish upper surface. The various populations sampled share identical ITS nr DNA sequences, of which the ITS2 regions are characterized by a unique secondary structure. Furthermore, we provide a detailed comparison of the characteristics of P. shennongjiana with morphologically similar species and a key to Peltigera species reported from China.

Species identification in the tropical lichen genus Phyllopsora is generally challenging and is based on ascospore morphology, vegetative dispersal units, thallus structure and secondary chemistry. As several type specimens are in poor condition and difficult to interpret, it is often unclear how these old names fit with the currently used taxonomy. In the present study, we aim to identify species boundaries in Phyllopsora s. str. supported by an integrative approach using multiple sources of evidence. We investigated a substantial amount of herbarium as well as freshly collected material and generated mtSSU and ITS sequence data from most of the described species, including several types. Species delimitation analyses are applied on the gene trees using mPTP and we construct a species tree of both markers with *BEAST, facilitating discussion of species delimitation and sister-relationships. Comparing morphology, chemistry and molecular data, we found that the mPTP analyses split established species repeatedly. Based on our integrative results, we exclude nine species from the genus, resurrect one (P. melanoglauca Zahlbr.), reduce two into synonymy with other Phyllopsora species and describe five as new to science: Phyllopsora amazonica Kistenich & Timdal (which shares the secondary chemistry (atranorin and terpenoid pattern) with P. halei chemotype 1, but differs, e.g., in having smaller areolae that are attached to a thinner, white prothallus, and in having more persistently marginate and less convex apothecia), Phyllopsora concinna Kistenich & Timdal (which shares the secondary chemistry (atranorin and parvifoliellin) with P. parvifoliella and P. rappiana, but differs from both in forming larger isidia, having a white prothallus, apothecial margin paler than the disc, and longer and broader ascospores), Phyllopsora furfurella Kistenich & Timdal (which is here segregated from P. furfuracea based on having a white prothallus and in containing skyrin in the hypothecium (K+ red)), Phyllopsora isidosa Kistenich & Timdal (which differs from P. byssiseda in forming a more crustose thallus with more delicate isidia, and from P. isidiotyla in forming somewhat coarser, less branched isidia) and Phyllopsora neotinica Kistenich & Timdal (a neotropical species here segregated from the now exclusively paleotropical P. chodatinica, differing in containing an unknown xanthone (not chodatin)). Lectotypes are designated for Biatora pyrrhomelaena Tuck., Lecidea leucophyllina Nyl., L. pertexta Nyl., and P. brachyspora Müll. Arg. In total, we accept 54 species in the genus Phyllopsora.

In this study we clarify the phylogeny and reassess the current taxonomy of the Micarea prasina group, focusing especially on the M. byssacea and M. micrococca complexes. The phylogeny was investigated using ITS, mtSSU and Mcm7 regions from 25 taxa belonging to the M. prasina group. A total of 107 new sequences were generated. Data were analyzed using maximum parsimony and maximum likelihood methods. The results reveal five undescribed well-supported lineages. Four of the lineages represent new species described as Micarea pseudomicrococca Launis & Myllys sp. nov., M. czarnotae Launis, van den Boom, Sérusiaux & Myllys sp. nov., M. microareolata Launis, Pykälä & Myllys sp. nov. and M. laeta Launis & Myllys sp. nov. In addition, a fifth lineage was revealed that requires further study. Micarea pseudomicrococca is characterized by an olive green granular thallus, small cream-white or brownish apothecia lacking the Sedifolia-grey pigment and two types of paraphyses up to 2 µm wide. Micarea czarnotae forms a granular, densely granular or continuous olive green thallus, convex to hemispherical apothecia often with the Sedifolia-grey pigment and no crystalline granules in the thallus. Micarea microareolata is characterized by a ± pale green areolate thallus (composed of goniocysts), cream-white apothecia lacking the Sedifolia-grey pigment and narrow spores. Micarea laeta has a vivid to olive green granular thallus, pale apothecia lacking the Sedifolia-grey pigment and wider spores compared to M. microareolata. Descriptions, images and a key are provided for the new species. Crystalline granules are introduced as a novel species-level character for Micarea.

Neoechinorhynchus is one of the most speciose genera of acanthocephalans, with approximately 116 described species. A recent study, aimed at establishing the genetic diversity of Neoechinorhynchus in Middle American freshwater fishes, validated nine species molecularly and morphologically and revealed the existence of 10 putative candidate species. Neoechinorhynchus golvani, a parasite commonly found in cichlids throughout Middle America with an allegedly large intraspecific morphological variability, was found to represent a species complex; species delimitation methods uncovered three additional genetic lineages. Here, we re-analyse the morphological and molecular data for N. golvani species complex infecting cichlids in that geographical area. A multivariate analysis of variance (MANOVA) was conducted particularly for the length of apical, middle and posterior hooks of the species/lineages of Neoechinorhynchus in cichlids, revealing morphological variation in the length of apical hooks for Lineage 8, although no morphological distinction was observed for Lineages 9 and 10. A new concatenated phylogenetic analysis of one mitochondrial and two ribosomal DNA genes was used to further corroborate the species delimitation among lineages; Neoechinorhynchus Lineage 8 was found to be morphologically and genetically distinct from its sister taxa, N. golvani and other two undescribed genetic lineages, and is formally described as a new species. Neoechinorhynchus costarricense n. sp. is described from the intestines of eight species of cichlids in Costa Rica. The new species is distinguished from the other species/lineages of Neoechinorhynchus in cichlids mainly by the size of the apical hooks of the proboscis.