Intraspecific Genetic Variation Research Articles

Molecular characterization and genetic diversity of three Stomoxys flies S. bengalensis, S. calcitrans, and S. sitiens (Diptera: Muscidae) in Central Thailand.

Stomoxys flies (Diptera: Muscidae) are hematophagous ectoparasites of medical and veterinary importance. In this study, three Stomoxys species, i.e. S. bengalensis, S. calcitrans, and S. sitiens, were collected from three provinces in Central Thailand with the aim of estimating the genetic divergence between species, for species identification, as well as within species, for a genetic diversity study based on the cytochrome c oxidase subunit I (COI) gene. Our results showed that the average intraspecific genetic divergences of Stomoxys flies ranged from 0.11% in S. sitiens to 0.98% in S. calcitrans, whereas the average interspecific genetic divergences ranged from 5.24% between S. sitiens and S. bengalensis to 6.69% between S. calcitrans and S. bengalensis. In addition, there was no overlap between the intraspecific and interspecific genetic divergences. The COI sequence analysis revealed a high haplotype diversity and low nucleotide diversity, reflecting a rapid population expansion after past bottlenecks. Moreover, there was no significant difference (P>0.05) in the pairwise population differentiation (Fst) among Stomoxys flies in Central Thailand, because of the lack of natural barriers, thus allowing genetic exchange between them. The monitoring of the haplotype network revealed that two lineages of S. calcitrans in Central Thailand were distributed in all study areas, including the Nakhon Pathom, Pathum Thani, and Saraburi Provinces. These findings may improve our understanding of the genetic patterns of these three Stomoxys flies, as well as the underlying biological mechanisms, which is knowledge that can be used for further effective control of these flies.

Species Identification and Mycotoxigenic Potential of Aspergillus Section Flavi Isolated from Maize Marketed in the Metropolitan Region of Asunción, Paraguay.

Zea mays var. amylacea and Zea mays var. indurata are maize ecotypes from Paraguay. Aspergillus section Flavi is the main spoilage fungus of maize under storage conditions. Due to its large intraspecific genetic variability, the accurate identification of this fungal taxonomic group is difficult. In the present study, potential mycotoxigenic strains of Aspergillus section Flavi isolated from Z. mays var. indurata and Z. mays var. amylacea that are marketed in the metropolitan region of Asunción were identified by a polyphasic approach. Based on morphological characters, 211 isolates were confirmed to belong to Aspergillus section Flavi. A subset of 92 strains was identified as Aspergillus flavus by mass spectrometry MALDI-TOF and the strains were classified by MALDI-TOF MS into chemotypes based on their aflatoxins and cyclopiazonic acid production. According to the partial sequencing of ITS and CaM genes, a representative subset of 38 A. flavus strains was confirmed. Overall, 75 A. flavus strains (86%) were characterized as producers of aflatoxins. The co-occurrence of at least two mycotoxins (AF/ZEA, FUM/ZEA, and AF/ZEA/FUM) was detected for five of the Z. mays samples (63%). Considering the high mycological bioburden and mycotoxin contamination, maize marketed in the metropolitan region of Asunción constitutes a potential risk to food safety and public health and requires control measures.

Pleistocene glaciations caused the latitudinal gradient of within-species genetic diversity.

Intraspecific genetic diversity is a key aspect of biodiversity. Quaternary climatic change and glaciation influenced intraspecific genetic diversity by promoting range shifts and population size change. However, the extent to which glaciation affected genetic diversity on a global scale is not well established. Here we quantify nucleotide diversity, a common metric of intraspecific genetic diversity, in more than 38,000 plant and animal species using georeferenced DNA sequences from millions of samples. Results demonstrate that tropical species contain significantly more intraspecific genetic diversity than nontropical species. To explore potential evolutionary processes that may have contributed to this pattern, we calculated summary statistics that measure population demographic change and detected significant correlations between these statistics and latitude. We find that nontropical species are more likely to deviate from neutral expectations, indicating that they have historically experienced dramatic fluctuations in population size likely associated with Pleistocene glacial cycles. By analyzing the most comprehensive data set to date, our results imply that Quaternary climate perturbations may be more important as a process driving the latitudinal gradient in species richness than previously appreciated.

Spineless and overlooked: DNA metabarcoding of autonomous reef monitoring structures reveals intra- and interspecific genetic diversity in Mediterranean invertebrates.

The ability to gather genetic information using DNA metabarcoding of bulk samples obtained directly from the environment is crucial to determine biodiversity baselines and understand population dynamics in the marine realm. While DNA metabarcoding is effective in evaluating biodiversity at community level, genetic patterns within species are often concealed in metabarcoding studies and overlooked for marine invertebrates. In the present study, we implement recently developed bioinformatics tools to investigate intraspecific genetic variability for invertebrate taxa in the Mediterranean Sea. Using metabarcoding samples from Autonomous Reef Monitoring Structures (ARMS) deployed in three locations, we present haplotypes and diversity estimates for 145 unique species. While overall genetic diversity was low, we identified several species with high diversity records and potential cryptic lineages. Further, we emphasize the spatial scale of genetic variability, which was observed from locations to individual sampling units (ARMS). We carried out a population genetic analysis of several important yet understudied species, which highlights the current knowledge gap concerning intraspecific genetic patterns for the target taxa in the Mediterranean basin. Our approach considerably enhances biodiversity monitoring of charismatic and understudied Mediterranean species, which can be incorporated into ARMS surveys.

First Discovery of the North American Leaf-Mining Moth Chrysaster ostensackenella (Lepidoptera: Gracillariidae) in Russia: The Genetic Diversity of a Novel Pest in Invaded vs. Native Range.

Here, we report the first detection of the North American leaf-mining moth Chrysaster ostensackenella (Fitch, 1859) (Lepidoptera: Gracillariidae) on North American black locust Robinia pseudoacacia (Fabaceae) in Primorsky Krai (the Russian Far East) in July 2022. Overall, six moths were reared from the leaf mines and identified based on adult morphology (forewing pattern and male genitalia) and three of them were DNA barcoding. Description of the leaf mines that allowed us to distinguish the damage of Ch. ostensackenella from other gracillariids associated with R. pseudoacacia is provided. The phylogeographic analysis comparing the DNA barcodes from Russia with those from other invaded countries in Europe (Italy) and East Asia (South Korea and Japan) and from the native range (North America) was performed. Intraspecific genetic diversity reached 3.29%. Altogether, 10 haplotypes were revealed among 21 studied specimens in the Holarctic. The detection of one haplotype common for Japan and the USA (North Carolina) suggests that the invasion to East Asia could have happened from the USA directly, rather than through Europe. A shared haplotype defined for Japan and the Russian Far East points at a possible moth species' spread to Primorsky Krai from earlier invaded Hokkaido. Further distribution of Ch. ostensackenella in East Asia and Europe is expected, bearing in mind the wide planting of R. pseudoacacia in these continents. Furthermore, an accidental introduction of the moth to the Southern Hemisphere, where black locust was introduced, is not ruled out.

Spatial prioritization of amphibian intraspecific genetic diversity: The need of accounting for palaeoenvironmental legacies

Biodiversity is a multifaceted concept, and conservation actions must consider all levels of biodiversity. Spatial prioritization of conservation efforts usually targets species diversity, while consideration of intraspecific genetic diversity is frequently hampered by the lack of data. Here, we combined genetic data (mtDNA) from multiple amphibian species and measures of environmental stability to i) identify the overall drivers of present-day intraspecific genetic diversity (nucleotide diversity and phylogenetic originality); ii) define priority areas for the conservation of genetic diversity, and iii) assess the surrogacy value of species diversity for genetic diversity in Italy and nearby areas. We tested for potential environmental predictors of genetic diversity and originality by fitting spatially-explicit Bayesian mixed models, and used species-specific predictions to generate spatial prioritizations of intraspecific genetic diversity. Present-day phylogenetic originality was positively correlated with climate and habitat stability since the Last Glacial Maximum (pseudo-R2: 0.61), while the same set of predictors had limited explanatory power for nucleotide diversity (pseudo-R2: 0.15). The spatial pattern of originality remarkably matched previously identified refugial areas for plants, as well as the distribution of some micro-endemic species. Prioritizations targeting phylogenetic originality showed that high conservation performance might be achieved with relatively low effort. However, the surrogacy between genetic and species-diversity was limited, implying that larger efforts would be needed to preserve genetic diversity while taking species diversity into account. Our study provides an example of how spatially-explicit approaches allow prioritization of conservation efforts accounting for both species and intraspecific genetic diversity, thus improving the long-term conservation of biodiversity.

SCoT markers efficiency for characterizing the intraspecific genetic diversity of Cyclamen coum, Helleborus caucasicus, Galanthus woronowii, Paeonia caucasica

Creating strategies of rare and endangered plants conservation requires studying the intraspecific genetic diversity of each particular species. Thus, efficient marker systems are necessary for implementing these objectives. In this study, the efficiency of 36 SCoT markers were evaluated for characterizing the intraspecific genetic diversity of Cyclamen сoum Mill., Helleborus caucasicus A. Brown, Galanthus woronowii Losinsk., Paeonia caucasica Schipcz. collected from different habitats of the Sochi National Park. Single markers, such as SCoT16 (58,2 %), SCoT6 (50,3 %), SCoT7 (49,6 %), SCoT8 (48,1 %), SCoT30 (42,8 %), SCoT29 (41,1 %), and SCoT20 (39,3 %) showed high levels of polymorphism, indicating good applicability of these markers to assess the intraspecific genetic diversity of the studied of the Western Caucasus flora species. In addition, the results of the study are important for the implementing conservation measures for these species and combating genetic erosion.

Diversity of Sand Snakes (Psammophiidae, Psammophis) in the Horn of Africa, with the description of a new species from Somalia

The biological diversity of the Horn of Africa is one of the least studied in the world. Yet the Horn supports rich communities of species that are mostly endemic to the region. Here we study the diversity of Sand Snakes (Psammophis) in East Africa, their phylogeny and systematics. Previous studies have unveiled several cryptic and potentially undescribed species of Psammophis that occur in the Horn and their taxonomic status has remained unclear to this day. We used sequence data from two mitochondrial and one nuclear genes to reconstruct the phylogeny of the genus, in which we included newly obtained samples of six different Psammophis species from Somalia, Ethiopia, Eritrea, Sudan, and Egypt. Our aim was to assess the status of some of the undescribed species, examine the level of intraspecific genetic variation within individual species, improve our understanding of the species distributions, and contribute to the taxonomy of the genus. Our results confirm the existence of two undescribed species, one in eastern Somalia, which we formally describe as new, and one in southern Ethiopia that we refer to as Psammophis cf. sudanensis in accordance with previous studies. Further, we provide first genetic data for the nominotypical subspecies of P. punctulatus and confirm the species status for its subspecies P. trivirgatus. In addition, we provide new genetic data for P. tanganicus from Ethiopia and Somalia, and range extension records for P. rukwae from Eritrea and Ethiopia and for P. aegyptius from Somalia. Our findings contribute considerably to our understanding of the diversity and distribution of Psammophis in East Africa.

Genetic variation and phylogenetic relationships of commercial populations of Bombus ignitus (Hymenoptera, Apidae) with wild populations in Eastern Asia

The bumblebee, Bombus ignitus (Hymenoptera, Apidae), plays a vital role in pollination in Northeast Asia, including Korea, China, Japan, and Far East Russia. Understanding the genetic makeup of the population can aid in its conservation. This study explores the DNA barcode region of cytochrome C oxidase subunit I (COI) of B. ignitus in commercial populations from Korea and Japan. The results reveal low intraspecific genetic diversity among commercially reared populations, with a maximum sequence divergence of 0.3%. Analysis of a 458-bp region of the COI gene, including 384 previously reported sequences, identified 20 haplotypes with the highest sequence divergence of 2.01% in East Asia. Commercial populations show a genetic similarity primarily with the Japanese population. Cross-mating with native populations could result in competition and genetic contamination, leading to reduced fitness and sensitivity to future environmental conditions. Morphological similarities make monitoring of such effects challenging. This study provides a basis for further research on population studies, conservation, and commercialization of local populations of B. ignitus for better pollination services while minimizing risks of reducing genetic diversity and increasing competition between native and introduced populations.

A molecular and ecological study of Macracanthorhynchus ingens (von Linstow, 1879) (Acanthocephala: Archiacanthocephala), in its paratenic and definitive hosts in southeastern Mexico and the Eastern USA.

The acanthocephalan Macracanthorhynchus ingens (von Linstow 1879) (Acanthocephala: Archiacanthocephala) is a parasite that infects the gut of carnivores (racoons, coyotes, wolves, foxes, badgers, skunks, opossum, mink and bears) as an adult and the body cavity of lizards, snakes, and frogs as a cystacanth in the Americas. In this study, adults and cystacanths of M. ingens from southeastern Mexico and southern Florida, USA, were identified morphologically by having a cylindrical proboscis armed with 6 rows of hooks each with 6 hooks. Hologenophores were used to sequence the small (SSU) and large (LSU) subunits of ribosomal DNA and cytochrome c oxidase subunit 1 (cox 1) from mitochondrial DNA.Phylogenetic analysis of the new SSU and LSU sequences of M. ingens placed them in a clade with other sequences available in GenBank identified as M. ingens. The cox 1 tree showed that the nine new sequences and six previously published sequences of M. ingens from the USA form a clade with other sequences previously identified as M. ingens from GenBank. The intraspecific genetic divergence among isolates from the Americas ranged from 0 to 2%, and in combination with the phylogenetic trees confirmed that the isolates belonged to the same species. The cox 1 haplotype network inferred with 15 sequences revealed 10 haplotypes separated from each other by a few substitutions. Rio Grande Leopard Frogs and Vaillant´s Frogs harbored cystacanths with low prevalence, 28% and 37% respectively, in Mexico. Brown Basilisks, an invasive lizard in Florida, USA, had high values of prevalence, 92% and 93% in males and females, respectively. Females harbored more cystacanths than males (0-39 vs 0-21) for unknown reasons that may, however, be related to ecological differences.

Mobilization of a novel diatom Mutator-Like Element (MULE) transposon inactivates the uridine monophosphate synthase (UMPS) locus in Phaeodactylum tricornutum.

Diatoms are photosynthetic unicellular microalgae that drive global ecological phenomena in the biosphere and are emerging sustainable feedstock for an increasing number of industrial applications. Diatoms exhibit enormous taxonomic and genetic diversity, which often result in peculiar biochemical and biological traits. Transposable elements (TE) represent a substantial portion of diatom genomes and have been hypothesized to exert a relevant role in enriching genetic diversity and centrally contribute to genome evolution. Here, through long-read whole genome sequencing, we identified a novel Mutator-Like Element (MULE) in the model diatom Phaeodactylum tricornutum, and we report the direct observation of its mobilization within the course of a single laboratory experiment. Under selective conditions, this novel TE inactivated the uridine monophosphate synthase (UMPS) gene of P. tricornutum, one of the few endogenous genetic loci currently targeted for selectable auxotrophy for functional genetics and genome editing applications. We report the observation of a recently mobilized transposon in diatoms with unique features. These include the combined presence of a MULE transposase with Zinc finger, SWIM-type domains, and of a diatom-specific E3 ubiquitin ligase of the zinc finger UBR type, which indicate a novel mobilization mechanism. Our findings provide new elements for the understanding of the role of TEs in diatom genome evolution and in the enrichment of intraspecific genetic variability.

Spatial Patterns of Neutral and Functional Genetic Variations along Dendritic Networks of Riverscape in Brown Trout Populations

Understanding how environmental gradients shape the spatial patterns of intraspecific genetic diversity is a central issue in ecological and evolutionary sciences. In riverine ecosystems, there is generally an increase in neutral genetic diversity downstream, as well as an increase in genetic differentiation among upstream populations. However, selective pressures may vary markedly along the upstream–downstream gradient, which could modify these patterns, but this has rarely been tested empirically. Here, we investigated how environmental gradients in a river network could shape the spatial patterns of intraspecific genetic diversity and differentiation in both neutral SNP markers and functional genetic markers putatively under natural selection (candidate SNPs associated with physiological functions and immune Major Histocompatibility Complex (MHC) loci) in wild brown trout populations. First, we showed that both the distance from the confluence and the centrality on the river network could explain the variation in genetic diversity and differentiation. Second, we found that both neutral and functional markers followed a similar pattern, with a higher genetic diversity and a lower genetic differentiation among populations that were more central and/or near to the confluence. This study highlights the importance of considering both the spatial and hydrological factors of a river network to understand and predict the role of dendritic connectivity in the spatial patterns of genetic diversity and differentiation in wild fish populations.

The unexplored effects of artificial selection on urban tree populations.

The unexplored effects of artificial selection on urban tree populations.

Genetic diversity varies with species traits and latitude in predatory soil arthropods (Myriapoda: Chilopoda).

To investigate the drivers of intra-specific genetic diversity in centipedes, a group of ancient predatory soil arthropods. Asia, Australasia and Europe. Present. Centipedes (Class: Chilopoda). We assembled a database of 1245 mitochondrial cytochrome c oxidase subunit I sequences representing 128 centipede species from all five orders of Chilopoda. This sequence dataset was used to estimate genetic diversity for centipede species and compare its distribution with estimates from other arthropod groups. We studied the variation in centipede genetic diversity with species traits and biogeography using a beta regression framework, controlling for the effect of shared evolutionary history within a family. A wide variation in genetic diversity across centipede species (0-0.1713) falls towards the higher end of values among arthropods. Overall, 27.57% of the variation in mitochondrial COI genetic diversity in centipedes was explained by a combination of predictors related to life history and biogeography. Genetic diversity decreased with body size and latitudinal position of sampled localities, was greater in species showing maternal care and increased with geographic distance among conspecifics. Centipedes fall towards the higher end of genetic diversity among arthropods, which may be related to their long evolutionary history and low dispersal ability. In centipedes, the negative association of body size with genetic diversity may be mediated by its influence on local abundance or the influence of ecological strategy on long-term population history. Species with maternal care had higher genetic diversity, which goes against expectations and needs further scrutiny. Hemispheric differences in genetic diversity can be due to historic climatic stability and lower seasonality in the southern hemisphere. Overall, we find that despite the differences in mean genetic diversity among animals, similar processes related to life-history strategy and biogeography are associated with the variation within them.

Genetic Diversity and Population Structure of Leishmania infantum in Morocco as Revealed by Multilocus Sequence Typing (MLST) Approach.

Leishmania infantum is endemic in Morocco, and it causes both visceral (VL) and cutaneous leishmaniasis (CL). In this study, the multilocus sequence typing (MLST) approach was used to investigate the phylogeny and population structure of Leishmania infantum strains isolated from CL and VL patients and the canine reservoir in different leishmaniasis endemic foci in Morocco. For this purpose, eight loci (pgm, alat, me, fh, g6pd, pgd, gpi and cytb) were amplified in 40 samples, out of which 31 were successfully sequenced. The genetic diversity analysis detected a high degree of intraspecific genetic variability among the studied strains. The phylogenetic and the haplotype analyses showed that most of the strains from the same geographical areas clustered together. The recombination among Leishmania infantum strains was revealed through a splits tree analysis and the number of recombination events. Moreover, the assessment of the gene flow between Leishmania infantum and Leishmania tropica through phylogenetic analysis and haplotype diversity in two endemic foci where the two species were sympatric showed no genetic exchange between the two species.

Detecting and analysing intraspecific genetic variation with eDNA: From population genetics to species abundance.

Advancements in environmental DNA (eDNA) approaches have allowed for rapid and efficient species detections in diverse environments. Although most eDNA research is focused on leveraging genetic diversity to identify taxa, some recent studies have explored the potential for these approaches to detect within-species genetic variation, allowing for population genetic assessments and abundance estimates from environmental samples. However, we currently lack a framework outlining the key considerations specific to generating, analysing and applying eDNA data for these two purposes. Here, we discuss how various genetic markers differ with regard to genetic information and detectability in environmental samples and how analysis of eDNA samples differs from common tissue-based analyses. We then outline how it may be possible to obtain species absolute abundance estimates from eDNA by detecting intraspecific genetic variation in mixtures of DNA under multiple scenarios. We also identify the major causes contributing to allele detection and frequency errors in eDNA data, discuss their consequences for population-level analyses and outline bioinformatic approaches to detect and remove erroneous sequences. This review summarizes the key advances required to harness the full potential of eDNA-based intraspecific genetic variation to inform population-level questions in ecology, evolutionary biology and conservation management.

Ocean crossers: A tale of disjunctions and speciation in the dwarf-fruticose Lichina (lichenized Ascomycota)

Lichens thrive in rocky coastal areas in temperate and cold regions of both hemispheres. Species of the genus Lichina, which form characteristic black fruiting thalli associated with cyanobacteria, often create distinguishable bands in the intertidal and supralittoral zones. The present study uses a comprehensive specimen dataset and four gene loci to (1) delineate and discuss species boundaries in this genus, (2) assess evolutionary relationships among species, and (3) infer the most likely causes of their current geographic distribution in the Northern and Southern hemispheres. A dated phylogeny describes the time frame in which extant disjunctions of species and populations were established. The results showed that the genus is integrated by four species, with Lichina pygmaea, L. confinis and the newly described L. canariensis from rocky seashores in the Canary Islands, occurring in the Northern Hemisphere, whereas L. intermedia is restricted to the Southern Hemisphere. Lichina intermedia hosted a much higher intraspecific genetic diversity than the other species, with subclades interpreted as species-level lineages by the different species delimitation approaches. However, a conservative taxonomic approach was adopted. This species showed a striking disjunct distribution between Australasia and southern South America. The timing for the observed interspecific and intraspecific divergences and population disjunctions postdated continental plate movements, suggesting that long-distance dispersal across body waters in the two hemispheres played a major role in shaping the current species distributions. Such ocean crossings were, as in L. canariensis, followed by speciation. New substitution rates for the nrITS of the genus Lichina were inferred using a tree spanning the major Ascomycota lineages calibrated using fossils. In conclusion, this work lays the foundation for a better understanding of the evolution through time and space of maritime lichens.

What Does “ITS” Say about Hybridization in Lineages of Sarsia (Corynidae, Hydrozoa) from the White Sea?

Hydrozoans are widely known for their complex life cycles. The life cycle usually includes an asexual benthic polyp, which produces a sexual zooid (gonophore). Here, we performed an extensive analysis of 183 specimens of the hydrozoan genus Sarsia from the White Sea and identified four types of gonophores. We also compared the type of gonophore with haplotypes of the molecular markers COI and ITS. Analysis of COI sequences recovered that the studied specimens related to the species S. tubulosa, S. princeps and S. lovenii, and that the S. lovenii specimens divided into two COI haplogroups. More intraspecific genetic diversity was revealed in the analysis of the ITS sequences. The Sarsia tubulosa specimens divided into two ITS haplotypes, and presumably, hybrid forms between these lineages were found. For S. lovenii, we identified 14 ITS haplotypes as a result of allele separation. Intra-individual genetic polymorphism of the ITS region was most likely associated with intraspecific crossing between the different haplotypes. The diversity of the morphotypes was associated with the genetic diversity of the specimens. Thus, we demonstrated that the morphologically variable species S. lovenii is represented in the White Sea by a network of intensively hybridizing haplotypes. Hybridization affects the morphology and maturation period of gonophores and presumably affects the processes of speciation.

DNA Barcoding of Black Flies (Diptera: Simuliidae) in Slovakia and Its Utility for Species Identification

DNA barcoding based on the cytochrome oxidase I gene is increasingly used in black flies (Diptera: Simuliidae), but extensive data for larger areas are still rare. Slovakia, with well-explored black fly fauna, was chosen to verify the reliability of DNA barcoding for species identification. The DNA barcoding region of the COI gene of 235 individuals of 25 black fly species from Slovakia was sequenced. Among them, 30 sequence clusters with assigned Barcode Index Numbers (BINs) were identified, and 5 of them were recorded for the first time. The average intraspecific genetic divergence was 0–3.24%, whereas the average interspecific divergence was 12.3–17.8%. Based on the barcode sequence, 14 species could be identified unambiguously, and 3 of them (Prosimulium latimucro, Simulium costatum, S. degrangei) are split into two or more barcode clusters. In eleven species, some degree of barcode sharing occurred, often combined with barcode splitting. The results showed hidden diversity as well as cases of shared barcode sequences among the studied species. Further investigation using other molecular markers is necessary due to the overlap of intraspecific and interspecific variability.

The Richness of Sarcocystis Species in the Common Gull (Larus canus) and Black-Headed Gull (Larus ridibundus) from Lithuania

The common gull (Larus canus) and the black-headed gull (Larus ridibundus) are common waterbird species in Lithuania. Until now, the composition of Sarcocystis species in these birds was unknown. The current study aimed to identify Sarcocystis spp. by the morphological examination of sarcocysts found in the muscle tissues and by DNA sequence analysis. Between 2011 and 2019, the leg muscles of 42 common gulls and 63 black-headed gulls were tested for Sarcocystis spp. Based on the methylene blue staining of squashed muscle samples, sarcocysts were detected in six common gulls (14.3%) and in six black-headed gulls (9.5%). Under a light microscope, one type of microcyst was observed. Sarcocysts were thread-like (2860–8250 × 40–180 μm) and had a smooth and thin (0.8–1.4 μm) cyst wall, while bradyzoites were banana-shaped and 5.0–9.2 × 1.3–2.4 μm in size. The sequencing of complete ITS1 showed the presence of S. columbae, S. halieti and S. wobeseri in the common gull and S. columbae and S. halieti in the black-headed gull. The highest intraspecific genetic variability was established for S. halieti, which is characterized by a wide host range. This species is considered to be pathogenic, therefore further histopathological examination of the various organs of gulls is needed.