Skip to main content

Molecular identification of four Sarcocystis species in the herring gull, Larus argentatus, from Lithuania



Birds of the family Laridae have not been intensively examined for infections with Sarcocystis spp. To date, sarcocysts of two species, S. lari and S. wobeseri, have been identified in the muscles of gulls. The aim of the present study was to evaluate the species richness of Sarcocystis in the herring gull, Larus argentatus, from Lithuania.


In the period between 2013 and 2019, leg muscles of 35 herring gulls were examined for sarcocysts of Sarcocystis spp. Sarcocystis spp. were characterised morphologically based on a light microscopy study. Four sarcocysts isolated from the muscles of each infected bird were subjected to further molecular examination. Sarcocystis species were identified by means of ITS1 sequence analysis.


Sarcocysts were detected in 9/35 herring gulls (25.7%). Using light microscopy, one morphological type of sarcocysts was observed. Sarcocysts were microscopic, thread-like, had a smooth and thin (about 1 µm) cyst wall and were filled with banana-shaped bradyzoites. On the basis of ITS1 sequences, four Sarcocystis species, S. columbae, S. halieti, S. lari and S. wobeseri, were identified. Furthermore, it was demonstrated that a single infected herring gull could host two Sarcocystis species indistinguishable under light microscopy.


Larus argentatus is the first bird species found to act as intermediate host of four Sarcocystis spp. According to current knowledge, five species, S. falcatula, S. calchasi, S. wobeseri, S. columbae and S. halieti can use birds belonging to different orders as intermediate hosts.


Protozoan parasites of the genus Sarcocystis are cyst-forming coccidians having an obligatory two-host prey-predator life-cycle [1, 2]. Asexual multiplication occurs in the intermediate host (IH), whereas sexual multiplication takes place in the small intestine of the definitive host (DH) [2, 3]. Thus far over 200 Sarcocystis species have been described; however, a much higher number or species diversity of these parasites is presumed [1, 4].

Birds may serve as intermediate or definitive hosts for many Sarcocystis species [1, 5,6,7,8]. More than 25 Sarcocystis species have been identified using birds as intermediate hosts [1, 9]. Two species, S. falcatula and S. calchasi are highly pathogenic for their intermediate hosts. Some species, such as S. falcatula, S. calchasi and S. wobeseri are not strictly specific to the intermediate host and could form sarcocysts in birds of several different orders [10,11,12,13]. By contrast, other species like S. fulicae, S. lari and S. ramphastosi are strictly specific to a single bird species [14,15,16].

Herring gulls are opportunistic predators of marine invertebrates, fishes, insects and birds, as well as opportunistic scavengers of dead animals and garbage [17, 18]. To date, only two Sarcocystis species, S. lari and S. wobeseri, have been described in birds of the family Laridae [15, 19]. The present study provides molecular identification of four Sarcocystis species from L. argentatus that are morphologically indistinguishable under light microscopy examination.


Collection of samples

A total of 35 herring gulls, road-killed and received from taxidermists between 2013 and 2019 were studied. Leg muscles were examined for the presence of sarcocysts.

Morphological analysis

The prevalence and intensity of infection with Sarcocystis spp. was evaluated in methylene blue-stained preparations. For this purpose, 28 oat-size fragments (about 1 g) of muscles were cut-off, stained with 0.2% methylene blue solution, clarified with 1.5% acetic acid solution and pressed in a glass compressor. After squeezing of fresh muscle tissues, sarcocysts were excised with the help of preparation needles and then morphologically characterized under a light microscope (LM).

DNA extraction and PCR

Four sarcocysts were extracted from the leg muscles of each infected bird and subjected to light microscopy and molecular investigation. For the molecular analysis, sarcocysts were placed in individual 1.5 ml tubes containing 20 μl of 96% ethanol and kept at − 20 °C. Genomic DNA was extracted from individual sarcocysts using the GeneJET Genomic DNA Purification Kit (Thermo Fisher Scientific Baltics, Vilnius, Lithuania) according to the manufacturer’s recommendations.

The complete ITS1 region was amplified using the SU1F/5.8SR2 primer pair [20]. Each PCR mixture consisted of 25 μl, containing 12.5 μl of Dream-Taq PCR Master Mix (Thermo Fisher Scientific, Waltham, US), 0.5 μM of each primer, 0.02 μg template DNA and nuclease-free water. The cycling conditions began with one cycle at 95 °C for 5 min followed by 35 cycles of 94 °C for 45 s, 60 °C for 60 s and 72 °C for 80 s, and a final extension step at 72 °C for 7 min. PCR products were evaluated using a 1.5% agarose gel and visualized via UV light after staining with 0.05 µg/ml ethidium bromide. Amplified DNA fragments were purified with exonuclease ExoI and alkaline phosphatase FastAP (Thermo Fisher Scientific).

DNA sequencing, sequence alignment and phylogenetic analysis

Sequencing reactions were performed using the Big-Dye Terminator v3.1 Cycle Sequencing Kit and the 3500 Genetic Analyzer (Applied Biosystems, Foster City, California, USA) according to the manufacturer’s recommendations. PCR products were sequenced directly using the PCR forward and reverse primers. The ITS1 sequences obtained in this study were compared with those of various Sarcocystis spp. using the Nucleotide BLAST program (megablast option). Sequences were aligned using the MUSCLE algorithm implemented in MEGA7 [21] software. The TOPALi v2.5 software [22] was used to select a nucleotide substitution model with the best fit to the aligned sequence dataset and to construct the phylogenetic tree under the Bayesian inference. Sequences for Sarcocystis spp. from L. argentatus generated in the present study are deposited in the GenBank database under the accession numbers MN450338-MN450373.


Sarcocysts were detected in 9 out of 35 (25.7%) herring gulls examined in Lithuania. The infection intensity of Sarcocystis spp. sarcocysts in 1 g of the leg muscle in L. argentatus varied from 1 to 85 cysts (mean = 33.0, median = 19.0). Examination of 36 sarcocysts under LM revelaed that they are morphologically similar. Sarcocysts were microscopic, thread-like, 2860–7930 × 43–200 μm in size, with a thin (0.7–1.5 μm), apparently smooth cyst wall. Septa divided sarcocysts into compartments filled with banana-shaped bradyzoites, 5.5–9.0 × 1.2–2.4 μm in size.

Surprisingly, the comparison of ITS1 sequences showed that the morphologically similar sarcocysts belonged to four different species of Sarcocystis, S. columbae, S. halieti, S. lari and S. wobeseri (Fig. 1). In the phylogenetic tree, the examined Sarcocystis spp. were placed into single-species clusters with a maximum support value. Based on ITS1, 833-bp long sequences of S. columbae obtained from L. argentatus (GenBank: MN450338-MN450339) demonstrated 99.9–100% identity with those of S. columbae (GenBank: GU253885, HM125052) from the wood pigeon (Columba palumbus). The BLAST analysis revealed that 860-bp long sequences of S. lari from L. argentatus (MN450357-MN450364) shared 99.1–100% identity with those of S. lari from the black-backed gull (Larus marinus) (IH, GenBank: JQ733510) and from the white-tailed sea eagle (Haliaeetus albicilla) (DH, GenBank: MF946597-MF946609). The 844-bp long ITS1 sequences of S. wobeseri obtained in this study (MN450365-MN450373) showed 99.8–100% identity with other sequences of S. wobeseri from the mallard duck (Anas platyrhynchos) (GenBank: JN256121), the barnacle goose (Branta leucopsis) (GenBank: GU475111) and L. argentatus (GenBank: HM159421). At ITS1, 830-bp long sequences of S. halieti from L. argentatus (MN450340-MN450356) shared 98.1–100% identity with other sequences of S. halieti from the great cormorant (Phalacrocorax carbo) (IH; GenBank: MH130209, JQ733513) and H. albicilla (DH; GenBank: MF946589-MF946596).

Fig. 1

Phylogenetic tree of selected Sarcocystis species based on ITS1 sequences. The figures next to branches show the posterior probability support values. Sequences generated in the present study are indicated with squares

Two Sarcocystis species identified in the present study, S. columbae (n = 2) and S. lari (n = 8) did not show any intraspecific genetic variability. The obtained ITS1 sequences of S. wobeseri differed only by one SNP (A/G) at nucleotide position 120, whereas S. halieti sequences demonstrated 98.7–100% identity. Thirteen identical sequences of S. halieti (MN450344-MN450356) showed 98.6% (MN450340-MN450341) and 98.7% (MN450342-MN450343) identity with other sequences obtained in the present study; sequences N450340-MN450341 differed in three SNPs from MN450342-MN450343.

Based on ITS1 sequences, S. columbae was identified in one out of nine infected birds. Two other species, S. lari and S. wobeseri, were confirmed in two and three herring gulls, respectively; whereas the most common species, S. halieti, was observed in five birds. It should be emphasized, that two different Sarcocystis species were discovered in L. argentatus (No. 9) and (No. 26). Larus argentatus (No. 9) harboured S. columbae and S. wobeseri, while L. argentatus (No. 26) had sarcocysts of S. halieti and S. wobeseri (Table 1).

Table 1 Sarcocystis species diversity in nine herring gulls from Lithuania based on molecular identification of four sarcocysts from each bird

The morphological analysis of sarcocysts isolated from herring gulls indicated that S. columbae, S. halieti, S. lari and S. wobeseri are indistinguishable based on the size of sarcocysts and bradyzoites, as well as the thickness of the sarcocyst wall (Table 2). For instance, S. wobeseri had the thickest sarcocysts wall and S. columbae was distinguished by the thinnest cyst wall. However, morphological parameters of the four Sarcocystis species overlapped and it was impossible to discriminate these parasites under LM.

Table 2 Morphological characteristics of Sarcocystis species from herring gulls


In the present study four Sarcocystis species, S. columbae, S. halieti, S. lari and S. wobeseri, were identified in L. argentatus from Lithuania. These species had thread-like sarcocysts with a smooth cyst wall and were indistinguishable from one another under LM. Previously two Sarcocystis species were recorded in gulls: S. wobeseri was detected in L. argentatus [19] and S. lari was described based on material from L. marinus [15]. To our knowledge, S. columbae and S. halieti are detected in gulls for the first time in our study. Sarcocysts of Sarcocystis sp. detected in the muscles of the California gull (Larus californicus) from Canada had a thin (0.8 μm) and smooth cyst wall [5]. In Kazakhstan, Pak & Eshtokina [23] discovered sarcocysts with a thin and smooth cyst wall and banana-shaped bradyzoites in the black-headed gull (L. ridibundus) and the common gull (L. canus). Thus, the morphology of sarcocysts observed in the gulls from Canada and Kazakhstan is quite similar to those recorded in the present study.

The results of the present study indicate that not only S. falcatula, S. calchasi and S. wobeseri [10,11,12,13] but also S. columbae and S. halieti could form sarcocysts in birds belonging to different orders. Sarcocysts of S. columbae have previously been detected in the woodpigeon C. palumbus (Columbiformes) and S. halieti has been detected in P. carbo (Suliformes) [11, 24]. Haliaeetus albicilla and the Eurasian sparrow hawk (Accipiter nisus) have been confirmed as definitive hosts for S. halieti [6, 25]. Accipiter nisus does not prey on adult great cormorants and mainly feeds on small passerines [26]. Consequently, the range of the intermediate hosts of S. halieti might be much wider, whereas S. lari has been identified only in gulls, in L. marinus and in L. argentatus so far. Hence, further studies are needed to reveal the intermediate host specificity of avian Sarcocystis species.

Sarcocystis species richness detected in L. argentatus in the present study is greater than that found in other bird species. Anas platyrhynchos serves as an intermediate host for three Sarcocystis species, S. anasi, S. rileyi and S. wobeseri [10, 15, 27,28,29,30]. According to current knowledge, other birds can be involved as intermediate hosts for one or two Sarcocystis species [14,15,16, 31, 32]. The richness of Sarcocystis species observed in L. argentatus can be related to the wide geographical distribution and great variety of feeding habitats of this bird species, where herring gulls might ingest sporocysts shed by the definitive hosts [33, 34]. It should be noted, that breeding colonies of L. argentatus are often located in the areas that are also used by other gull species, cormorants and ducks, acting as intermediate hosts of S. lari, S. halieti and S. wobeseri, respectively [35].

The morphology of the sarcocysts wall is the main diagnostic feature for morphological separation of Sarcocystis species in intermediate hosts [1]. Under LM, a thin and smooth sarcocyst wall was described for several avian Sarcocystis species, S. calchasi, S. columbae, S. corvusi, S. halieti, S. fulicae, S. lari and S. wobeseri. These species also share similar sarcocyst wall structure under the transmission electron microscope [10, 11, 15, 16, 36, 37]. Thus, Sarcocystis species discussed are apparently morphologically indistinguishable. To the best of our knowledge, our study provides first evidence for several Sarcocystis spp. with a very similar morphological appearance under LM using a single bird species as an intermediate host. We have also demonstrated that one bird might host two Sarcocystis species, which could not be distinguished under LM. It should be emphasized, that the conclusions about Sarcocystis spp. richness in certain bird species might be misleading if only one sarcocyst is isolated for molecular identification. In 2011, our research group detected sarcocysts in the neck and leg muscles of four out of 11 herring gulls examined [19]. Under LM, one morphological type of sarcocyst was observed and only one excised cyst was subjected to molecular examination. At that time, it was assumed that sarcocysts detected in four herring gulls belonged to S. wobeseri. In contrast, the present study showed that L. argentatus can act as an intermediate host for four Sarcocystis species. Hence, when seeking to determine Sarcocystis species richness in birds, several sarcocysts should be isolated from each infected individual.


In the present study four Sarcocystis species, S. columbae, S. halieti, S. lari and S. wobeseri were identified in L. argentatus from Lithuania by means of ITS1 sequence analysis. Detected Sarcocystis species were morphologically indistinguishable under LM. In comparison with other bird species, L. argentatus has the highest Sarcocystis species richness. The results of the present study showed that S. columbae and S. halieti could use birds of different orders as intermediate hosts. It was revealed that muscles of a single herring gull could be infected with two Sarcocystis species indistinguishable under LM; therefore, in order to determine Sarcocystis species richness in bird intermediate hosts, or at least within genus Larus, we recommend molecular characterization of several sarcocysts isolated from each infected individual.

Availability of data and materials

Data supporting the conclusions of this article are included within the article. The ITS1 sequences generated in the present study were submitted to the GenBank database under the accession numbers MN450338-MN450373.



light microscopy


internal transcribed spacer


single nucleotide polymorphism


intermediate host


definitive host




  1. 1.

    Dubey JP, Calero-Bernal R, Rosenthal BM, Speer CA, Fayer R. Sarcocystosis of animals and humans. 2nd ed. Boca Raton: CRC Press; 2016.

    Google Scholar 

  2. 2.

    Prakas P, Butkauskas D. Protozoan parasites from genus Sarcocystis and their investigations in Lithuania. Ekologija. 2012;58:45–58.

    Article  Google Scholar 

  3. 3.

    Odening K, Frölich K, Kirsch B, Bockhardt I. Sarcocystis: development of sporocysts in cell culture. Anim Behav. 1997;7:9–16.

    Google Scholar 

  4. 4.

    Olias P, Olias L, Lierz M, Mehlhorn H, Gruber AD. Sarcocystis calchasi is distinct to Sarcocystis columbae sp. nov. from the wood pigeon (Columba palumbus) and Sarcocystis sp. from the sparrowhawk (Accipiter nisus). Vet Parasitol. 2010;171:7–14.

    Article  Google Scholar 

  5. 5.

    Drouin TE, Mahrt JL. The prevalence of Sarcocystis Lankester, 1882, in some bird species in western Canada, with notes on its life cycle. Can J Zool. 1979;57:1915–21.

    CAS  Article  Google Scholar 

  6. 6.

    Gjerde B, Vikøren T, Hamnes IS. Molecular identification of Sarcocystis halieti n. sp., Sarcocystis lari and Sarcocystis truncata in the intestine of a white-tailed sea eagle (Haliaeetus albicilla) in Norway. Int J Parasitol Parasites Wildl. 2018;7:1–11.

    Article  Google Scholar 

  7. 7.

    Munday BL, Hartley WJ, Harrigan KE, Presidente PJ, Obendorf DL. Sarcocystis and related organisms in Australian wildlife: II. Survey of findings in birds, reptiles, amphibians and fish. J Wildl Dis. 1979;15:57–73.

    CAS  Article  Google Scholar 

  8. 8.

    Černá Ž. The role of birds as definitive hosts and intermediate hosts of heteroxenous coccidians. J Protozool. 1984;1984(31):579–81.

    Article  Google Scholar 

  9. 9.

    Odening K. The present state of species-systematics in Sarcocystis Lankester, 1882 (Protista, Sporozoa, Coccidia). Syst Parasitol. 1998;41:209–33.

    Article  Google Scholar 

  10. 10.

    Kutkienė L, Prakas P, Sruoga A, Butkauskas D. The mallard duck (Anas platyrhynchos) as intermediate host for Sarcocystis wobeseri sp. nov. from the barnacle goose (Branta leucopsis). Parasitol Res. 2010;107:879–88.

    Article  Google Scholar 

  11. 11.

    Prakas P, Butkauskas D, Sruoga A, Švažas S, Kutkienė L. Identification of Sarcocystis columbae in wood pigeons (Columba palumbus) in Lithuania. Vet Zootech. 2011;55:33–9.

    Google Scholar 

  12. 12.

    Konradt G, Bianchi MV, Leite-Filho RV, da Silva BZ, Soares RM, Pavarini SP, Driemeier D. Necrotizing meningoencephalitis caused by Sarcocystis falcatula in bare-faced ibis (Phimosus infuscatus). Parasitol Res. 2017;116:809–12.

    Article  Google Scholar 

  13. 13.

    Parmentier SL, Maier-Sam K, Failing K, Gruber AD, Lierz M. High prevalence of Sarcocystis calchasi in racing pigeon flocks in Germany. PLoS One. 2019;15:14.

    Google Scholar 

  14. 14.

    Dubey JP, Lane E, van Wilpe E. Sarcocystis ramphastosi sp. nov. and Sarcocystis sulfuratusi sp. nov. (Apicomplexa, Sarcocystidae) from the keel-billed toucan (Ramphastos sulfuratus). Acta Parasitol. 2004;49:93–101.

    Google Scholar 

  15. 15.

    Prakas P, Kutkienė L, Butkauskas D, Sruoga A, Žalakevičius M. Description of Sarcocystis lari sp. n. (Apicomplexa: Sarcocystidae) from the great black-backed gull, Larus marinus (Charadriiformes: Laridae), on the basis of cyst morphology and molecular data. Folia Parasitol. 2014;61:11–7.

    Article  Google Scholar 

  16. 16.

    Prakas P, Butkauskas D, Švažas S, Juozaitytė-Ngugu E, Stanevičius V. Morphologic and genetic identification of Sarcocystis fulicae n. sp. (Apicomplexa: Sarcocystidae) from the Eurasian Coot (Fulica atra). J Wildl Dis. 2018;54:765–71.

    Article  Google Scholar 

  17. 17.

    Kubetzki U, Garthe S. Distribution, diet and habitat selection by four sympatrically breeding gull species in the south-eastern North Sea. Mar Biol. 2003;143:199–207.

    Article  Google Scholar 

  18. 18.

    Nisbet ICT, Weseloh DV, Hebert EC, Mallory LM, Poole FA, Ellis CJ, Pyle P, Pattern AM. Herring Gull (Larus argentatus). 2017; Accessed 30 Aug 2019.

  19. 19.

    Prakas P, Kutkienė L, Sruoga A, Butkauskas D. Sarcocystis sp. from the herring gull (Larus argentatus) identity to Sarcocystis wobeseri based on cyst morphology and DNA results. Parasitol Res. 2011;109:1603–8.

    Article  Google Scholar 

  20. 20.

    Gjerde B. Molecular characterisation of Sarcocystis rileyi from a common eider (Somateria mollissima) in Norway. Parasitol Res. 2014;113:3501–9.

    Article  Google Scholar 

  21. 21.

    Kumar S, Stecher G, Tamura K. MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol Biol Evol. 2016;33:1870–4.

    CAS  Article  Google Scholar 

  22. 22.

    Milne I, Wright F, Rowe G, Marshall DF, Husmeier D, McGuire G. TOPALi: software for automatic identification of recombinant sequences within DNA multiple alignments. Bioinformatics. 2004;20:1806–7.

    CAS  Article  Google Scholar 

  23. 23.

    Pak SM, Eshtokina NV. Sarcosporidians of birds. Sarcosporidians of animals in Kazakhstan. Almaty: Nauka; 1984.

    Google Scholar 

  24. 24.

    Prakas P, Butkauskas D, Švažas S, Stanevičius V. Morphological and genetic characterisation of Sarcocystis halieti from the great cormorant (Phalacrocorax carbo). Parasitol Res. 2018;117:3663–7.

    Article  Google Scholar 

  25. 25.

    Mayr SL, Majer K, Müller J, Enderlein D, Gruber AD, Lierz M. Accipiter hawks (Accipitridae) confirmed as definitive hosts of Sarcocystis turdusi, Sarcocystis cornixi and Sarcocystis sp. ex Phalacrocorax carbo. Parasitol Res. 2016;115:3041–7.

    Article  Google Scholar 

  26. 26.

    Zawadska D, Zawadski J. Breeding populations and diets of the sparrowhawk Accipiter nisus and the hobby Falco subbuteo in the Wigry National Park (NE Poland). Acta Ornithol. 2001;36:25–31.

    Article  Google Scholar 

  27. 27.

    Kutkienė L, Sruoga A, Butkauskas D. Sarcocystis sp. from white-fronted goose (Anser albifrons): cyst morphology and life cycle studies. Parasitol Res. 2006;99:562–5.

    Article  Google Scholar 

  28. 28.

    Kutkienė L, Sruoga A, Butkauskas D. Sarcocystis sp. from the goldeneye (Bucephala clangula) and the mallard (Anas platyrhynchos): cyst morphology and ribosomal DNA analysis. Parasitol Res. 2008;102:691–6.

    Article  Google Scholar 

  29. 29.

    Kutkienė L, Prakas P, Sruoga A, Butkauskas D. Identification of Sarcocystis rileyi from the mallard duck (Anas platyrhynchos) in Europe: cyst morphology and results of DNA analysis. Parasitol Res. 2011;108:709–14.

    Article  Google Scholar 

  30. 30.

    Kutkienė L, Prakas P, Sruoga A, Butkauskas D. Description of Sarcocystis anasi sp. nov. and Sarcocystis albifronsi sp. nov. in birds of the order Anseriformes. Parasitol Res. 2012;110:1043–6.

    Article  Google Scholar 

  31. 31.

    El-Morsey A, El-Seify M, Desouky AY, Abdel-Aziz MM, El-Dakhly KM, Kasem S, et al. Morphologic and molecular characteristics of Sarcocystis atraii n. sp. (Apicomplexa: Sarcocystidae) infecting the common coot (Fulica atra) from Egypt. Acta Parasitol. 2015;60:691–9.

    Article  Google Scholar 

  32. 32.

    El-Morsey A, EL-Seify M, Desouky AY, Abdel-Aziz MM, Sakai H, Yanai T. Sarcocystis chloropusae (Protozoa Sarcocystidae) n sp from the common moorhen (Gallinula chloropus) from Egypt. Parasitology. 2015;142:1063–5.

    CAS  Article  Google Scholar 

  33. 33.

    Camphuysen CJ. A historical ecology of two closely related gull species (Laridae): multiple adaptations to a man-made environment. PhD thesis, University of Groningen, Groningen, Netherlands; 2013.

  34. 34.

    Somers MCH, Lozer MN, Quinn SJ. Interactions between double-crested cormorants and herring gulls at a shared breeding site. Waterbirds. 2007;30:241–50.

    Article  Google Scholar 

  35. 35.

    Marchowski D, Jankowiak Ł, Wysocki D. Newly demonstrated foraging method of herring gulls and mew gulls with benthivorous diving ducks during the nonbreeding period. Auk. 2015;133:31.

    Article  Google Scholar 

  36. 36.

    Olias P, Gruber AD, Hafez HM, Heydorn AO, Mehlhorn H, Lierz M. Sarcocystis calchasi sp nov of the domestic pigeon (Columba livia f. domestica) and the northern goshawk (Accipiter gentilis): light and electron microscopical characteristics. Parasitol Res. 2010;106:577–85.

    Article  Google Scholar 

  37. 37.

    Prakas P, Kutkienė L, Butkauskas D, Sruoga A, Žalakevičius M. Molecular and morphological investigations of Sarcocystis corvusi sp. nov. from the jackdaw (Corvus monedula). Parasitol Res. 2013;112:1163–7.

    Article  Google Scholar 

Download references


This study was supported by the Open Access research infrastructure of the Nature Research Centre under the Lithuanian open access network initiative. The authors are grateful to Valentinas Pabrinkis (Nature Research Centre, Vilnius, Lithuania) who provided samples for the study.


Not applicable.

Author information




PP conceived and designed the laboratory tests. EJN performed the experiments. PP and DB contributed reagents/materials/analysis tools. PP, DB and EJN drafted the manuscript. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Petras Prakas.

Ethics declarations

Ethics approval and consent to participate

Not applicable. Samples were collected with the permission of the Ministry of Environment of the Republic of Lithuania (2013-03-26 no.14; 2014-03-03 no.15; 2017-03-23 no.26-A4-3119; 2019-03-01 no. 26-A4-1535).

Consent for publication

Not applicable.

Competing interests

The authors declare that they have no competing interests.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit The Creative Commons Public Domain Dedication waiver ( applies to the data made available in this article, unless otherwise stated in a credit line to the data.

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Prakas, P., Butkauskas, D. & Juozaitytė-Ngugu, E. Molecular identification of four Sarcocystis species in the herring gull, Larus argentatus, from Lithuania. Parasites Vectors 13, 2 (2020).

Download citation


  • Sarcocystis
  • Herring gull
  • ITS1
  • Species differentiation