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Protozoans of genus Sarcocystis are widespread parasites infecting mammals, birds, and reptiles. Morphology of their sarcocysts is an important criterion for species identification. However, as more and more morphologically similar Sarcocystis species are being found and described, additional methods for their routine diagnostics are needed. We investigated restriction fragment length polymorphism (RFLP) as potential alternative to sequencing data analysis for the identification of Sarcocystis species using birds as an intermediate host. The internal transcribed spacer 1 (ITS1) region sequences of seventeen Sarcocystis species (S. albifronsi, S. anasi, S. calchasi, S. columbae, S. cornixi, S. corvusi, S. cristata, S. halieti, S. falcatula, S. fulicae, S. kutkienae, S. lari, S. lindsayi, S. rileyi, S. turdusi, S.wenzeli, and S. wobeseri) of interest were analysed and five best-fitting endonucleases generating most informative restriction fragments were selected for routine testing. In general, RFLP analyses are always inconclusive as they target very short DNA sequences. However, it can be an irreplaceable technique when fast and cheap identification and discrimination of known species are required, which was our main goal and preliminary results indicate that RFLP could be successfully used when identifying closely related avian Sarcocystis species with just two nucleases. [ABSTRACT FROM AUTHOR]

Fusarium head blight (FHB) is an important crop disease worldwide and is mainly caused by members of the Fusarium graminearum species complex. F. graminearum sensu stricto is the most common cosmopolitan and predominant FHB causal agent in Europe. Thus far, the majority of studies have focused on the primary hosts (wheat and barley) of this pathogen, while the relationships between other sources of infection remain unclear. We monitored and sampled test fields over the course of 3 years and acquired 804 F. graminearum isolates from different sources: primary hosts and other plant species included in the crop rotations, weeds from the test fields, decaying plant residue, soil samples, and crop seed. Of these isolates, 73.3% had the 15-acetyldeoxynivalenol genotype and 26.7% had the 3-acetyldeoxynivalenol genotype. F. graminearum isolation rates from weeds (>50%) were much higher than from soil (< 10%) or decaying plant matter (4%). Variable number of tandem repeat markers were used for population analysis. Noticeable genetic differentiation was detected between isolates from living plants and soil biome. In contrast, absence of any noticeable division between primary and alternative plant host communities indicates the importance of weeds and other segetal plants for FHB control and prevention. [ABSTRACT FROM AUTHOR]

Our previous studies reported that phytoplasma was the causative agent of the pine disease in the Curonian Spit, Lithuania. In this study, insects from diseased pine trees and their adjacent areas were collected from 2016 to 2019 to further identify potential insect vectors that spread phytoplasmas. A total of 1018 phloem-feeding insects (order Hemiptera) were identified, 98.62% of which were aphids (Aphididae), and no known phytoplasma vectors were found. The results from semi-nested polymerase chain reaction (PCR) using phytoplasma-specific universal primers revealed that phytoplasmas were detected in Scots pine aphids (Cinara pini (Linnaeus, 1758)), waxy grey pine needle aphids (Cinara pineti (Fabricius, 1781)), and species-unknown aphids. Further sequence analysis and virtual RFLP (restriction fragment length polymorphism) analysis of aphid-harbored phytoplasma strains indicated that they were closely related to 'Candidatus Phytoplasma pini' (16SrXXI-A), but mainly 16SrXXI-A variants, which were also main strains identified in diseased pine trees. In addition, three new phytoplasma subgroups were delineated in the present study. Subgroups 16SrXXI-C and 16SrXXI-D were unveiled from previously identified (but classification was overlooked) Lithuanian pine phytoplasma strains. Subgroup 16SrXXI-E was discovered from the newly identified aphid-harbored phytoplasmas. Further transmission trial studies on these aphids will provide insights into the epidemiology and pathosystem of pine phytoplasma diseases, as well as disease management. [ABSTRACT FROM AUTHOR]

Simple Summary: Western chimpanzees are currently listed as a Critically Endangered subspecies. Human encroachment has taken a toll on this great ape due to fragmented habitat and the exchange of pathogens. This epidemiological study investigated the occurrence and genetic diversity of intestinal and blood parasites in faecal samples from wild chimpanzees living in the Dindefelo Community Nature Reserve, Senegal. We paid special attention to potential human-driven sources of infection and transmission pathways. Potential diarrhoea-causing protist parasites (e.g., Cryptosporidium spp., Giardia duodenalis, Entamoeba histolytica) were detected at low infection rates (and densities) or absent, whereas commensals (Entamoeba dispar) or protist of uncertain pathogenicity (Blastocystis sp.) were far more abundant. We detected Sarcocystis spp. in chimpanzee faeces. Blood protist parasites such as Plasmodium spp. and Trypanosoma brucei spp. (the etiological agents of malaria and sleeping sickness, respectively, in humans) were also found at low prevalences, but microfilariae of the nematode Mansonella perstans were frequently found. Molecular analyses primarily revealed host-adapted species/genotypes and an apparent absence of gastrointestinal clinical manifestations in infected chimpanzees. Zoonotic events of still unknown frequency and directionality may have taken part between wild chimpanzees and humans sharing natural habitats and resources. Wild chimpanzee populations in West Africa (Pan troglodytes verus) have dramatically decreased as a direct consequence of anthropogenic activities and infectious diseases. Little information is currently available on the epidemiology, pathogenic significance, and zoonotic potential of protist species in wild chimpanzees. This study investigates the occurrence and genetic diversity of intestinal and blood protists as well as filariae in faecal samples (n = 234) from wild chimpanzees in the Dindefelo Community Nature Reserve, Senegal. PCR-based results revealed the presence of intestinal potential pathogens (Sarcocystis spp.: 11.5%; Giardia duodenalis: 2.1%; Cryptosporidium hominis: 0.9%), protist of uncertain pathogenicity (Blastocystis sp.: 5.6%), and commensal species (Entamoeba dispar: 18.4%; Troglodytella abrassarti: 5.6%). Entamoeba histolytica, Enterocytozoon bieneusi, and Balantioides coli were undetected. Blood protists including Plasmodium malariae (0.4%), Trypanosoma brucei (1.3%), and Mansonella perstans (9.8%) were also identified. Sanger sequencing analyses revealed host-adapted genetic variants within Blastocystis, but other parasitic pathogens (C. hominis, P. malariae, T. brucei, M. perstans) have zoonotic potential, suggesting that cross-species transmission between wild chimpanzees and humans is possible in areas where both species overlap. Additionally, we explored potential interactions between intestinal/blood protist species and seasonality and climate variables. Chimpanzees seem to play a more complex role on the epidemiology of pathogenic and commensal protist and nematode species than initially anticipated. [ABSTRACT FROM AUTHOR]

In order to devise a method for rapid detection of 'Candidatus (Ca.) Phytoplasma pini' and for distinguishing it rapidly from other phytoplasmas, we carried out preliminary sequencing of Lithuanian 'Ca. Phytoplasma pini' strain PineBL2 using Illumina (NGS) technology and targeted sequencing employing universal phytoplasma primers. We focused on two resulting chromosomal segments that contained a 16S rRNA gene and a translation elongation factor EF‐TU gene (tuf), respectively. Based on alignments of the 'Ca. Phytoplasma pini' gene sequences with the corresponding sequences of other phytoplasmas, we designed new primer pairs for PCR‐based detection of 'Ca. Phytoplasma pini'. Because 'Ca. Phytoplasma pini' strains are expected to reside in the pine phloem in a very low titre, one might expect that they could be detected only by nested PCR. By contrast, the primers and PCR protocols designed in the current work enabled rapid direct PCR detection and identification of 'Ca. Phytoplasma pini' by amplifying a 484 bp 16S rDNA segment and a 513 bp tuf gene fragment that contain regions unique to this phytoplasma. [ABSTRACT FROM AUTHOR]