Search

The monogeneric family Calomyscidae includes the brush-tailed mice, genus Calomyscus , which have a Palearctic distribution ranging from the Hindu Kush Mountains in western Pakistan to the eastern Mediterranean region. Zagros Mountains—stretching from northwest to south of Iran—was assumed for long as the range of a single species, Calomyscus bailwardi ; however, recent studies revealed considerable heterogeneities among the geographical populations presumably referring to the persistence of undescribed diversity. One such group from the western Zagros Mountains has recently been recognized as C. behzadi Akbarirad, Dezhman, Aliabadian, Siahsarvie, Shafaeipour, and Mirshamsi, 2021. Using molecular, morphometric, geometric morphometric, and karyotypic data, we examine the divergence of 2 additional groups from western Iran and name these as new species. The first of these species is distributed in the northern Zagros and western Alborz Mountains in northwestern Iran was recovered as the sister species to C. urartensis from the South Caucuses. The second species from a single locality in the central Zagros Mountains was clustered closely with C. grandis from the western Alborz Mountains. Morphologic analyses, karyological features, and genetic distances disentangled these 2 new species from their sister species. [ABSTRACT FROM AUTHOR]

Haemosporidian parasites that infect birds (Apicomplexa: Haemosporida) are blood parasites that require an invertebrate host (vector) and a vertebrate host for their lifecycle and cause malaria-like diseases. This group of parasites has provided valuable insights into host specificity, virulence, and parasite dispersal. Additionally, they have played a significant role in reshaping our understanding of the evolutionary history of apicomplexans. In order to accurately identify species and to address phylogenetic questions such as the timing of the haemosporidian radiation, the use of a sufficiently large genetic data set is crucial. However, acquiring this genetic data poses significant challenges. In this research, a sensitive nested PCR assay was developed. This assay allows for the easy amplification of complete mitochondrial genomes of haemosporidian parasites in birds, even during the chronic stage of infection. The effectiveness of this new nested PCR assay was evaluated using blood and tissue samples of birds with verified single parasite infections from previous studies. The approach involves amplifying four overlapping fragments of the mitochondrial genome and requires DNA extracts from single-infected samples. This method successfully amplified the complete mitochondrial genomes of 24 distinct haemosporidian parasite lineages found in various bird species. This data is invaluable for conducting phylogenetic analyses and accurately defining species. Furthermore, this study proposes the existence of at least 15 new haemosporidian parasite species based on the genetic information obtained. Data regarding pGRW04, previously categorized as Plasmodium relictum like pSGS1 and pGRW11, indicates that the pGRW04 lineage is actually a separate, hidden Plasmodium species. [ABSTRACT FROM AUTHOR]

Ants of the «Formica picea—F. candida» complex are widespread across Eurasia. However, it is still a matter of debate if it constitutes one or two species. In this study, we collected a sample of specimens from different parts of Eurasia, sequenced the mitochondrial cox1 and cytb genes, as well as three nuclear loci—wg, Top1, and ITS2—and analyzed the available published data. We found this complex to contain a new, yet undescribed, taxon that has a large distribution in Siberia and East Asia. Thus, the «Formica picea—F. candida» complex consists of at least three taxa with distinct distributions. [ABSTRACT FROM AUTHOR]

In this work, for the first time, the genetic variability of the Metagonimus suifunensis population in the Russian southern Far East was estimated based on the full-length sequences of the nad1 gene of mitochondrial DNA. In addition, for a sample of the same size, the sequences of cox1 and cytb genes, previously used for population studies for M. suifunensis, were reanalysed. Three markers were combined to a common sequence, and the obtained data were studied. Despite the higher level of variability, nad1 and cox1 mtDNA genes did not reveal subdivisions within the population. The combined dataset made it possible to determine that the sample from the Odyr River was the centre of the species' range formation and clarified the continental migration route of the parasite from south to north. According to the data obtained, it was presumed that piscivorous birds participate in the life cycle of the parasite. The subdivision within population revealed that using all three mitochondrial markers is consistent with the features of differentiation within populations of related species, but the reasons for its formation remain unclear due to the insufficient amount of data and the use of different markers in studies of different species. [ABSTRACT FROM AUTHOR]

The largest genus in the Sylviidae family is Sylvia, which is the archetype for warblers. It contains up to 28 species that are distributed in the Old World. Here, we study Iranian Sylvia using cytochrome c oxidase subunit 1 (cox1) and cytochrome b (cytb) mitochondrial markers to identify them and place them in a phylogenetic context using all available GenBank samples of sufficient length. Bayesian trees for both cytb and cox1 mitochondrial markers are compatible. Previous results suggesting that the Sylvia curruca complex is separated into two major clades with high support are corroborated, as our results show two main clades, one that includes Sylvia atricapilla and Sylvia borin plus Pseudoalcippe atriceps and Lioptilus nigricapillus, and the remainder of the Sylvia species in the other one. Our study did not support the geographical groups: The West-Mediterranean group, the Central-Mediterranean group, and the East-Mediterranean or the African-European species, the African-Asian species, and the Mediterranean species inferred on molecular evidenced by previous studies. Sylvia cantillans is divided into three clades, corroborating previous studies. All Iranian samples were conclusively identified to species level, and none of them give rise to any further taxonomic inquiry.

Ants of the «Formica picea—F. candida» complex are widespread across Eurasia. However, it is still a matter of debate if it constitutes one or two species. In this study, we collected a sample of specimens from different parts of Eurasia, sequenced the mitochondrial cox1 and cytb genes, as well as three nuclear loci—wg, Top1, and ITS2—and analyzed the available published data. We found this complex to contain a new, yet undescribed, taxon that has a large distribution in Siberia and East Asia. Thus, the «Formica picea—F. candida» complex consists of at least three taxa with distinct distributions.

The common vole Microtus arvalis (Pallas, 1778) is the most widespread Microtus species. It has two forms - the European arvalis form (2n: 46, NF: 84) and the Asian obscurus form (2n: 46, NF: 72). The present study aimed to clarify the taxonomic status of M. arvalis populations distributed in Eastern Turkey, Europe and Asia by analysing two mitochondrial (CYTB and COX1) and one nuclear (IRBP) markers. Phylogenetic dendrograms (median-joining networks and Bayesian trees) constructed using the mitochondrial markers clearly separated the Anatolian population from the European and Asian populations. Contrarily, any explicit differentiation was not shown in IRBP analyses. Mean and net genetic distance values (d) were found to be notably low for three markers. Species delimitation test (Automatic Barcode Gap Discovery Method) supported these results. Our results indicate that the arvalis and obscurus forms are not sufficiently differentiated to be considered different species, while the Anatolian population has only recently split from the Asian population. Together, these findings demonstrate that the speciation process is ongoing. [ABSTRACT FROM AUTHOR]

Babesia gibsoni is considered as an emerging protozoan parasite of dogs in North America and Europe. However, no data have been published on its prevalence, molecular-phylogenetic characteristics and associated co-infections in dogs used for illegal fighting (i.e. predisposed to acquiring this piroplasm via biting) in Europe. In this study, blood samples from 79 American Staffordshire Terrier dogs, confiscated for illegal dog fights, were molecularly analyzed for tick-borne pathogens. Babesia gibsoni was detected in 32 dogs, i.e. with a prevalence of 40.5%. In addition, Babesia vulpes was found in 8 samples (prevalence of 10.1%), for the first time in dogs in Hungary. Canine hemoplasmas were also identified in 49 samples (62%): only Mycoplasma haemocanis in 32 (40.5%) dogs, only “Candidatus Mycoplasma haematoparvum” in 9 (11.4%) dogs, and both hemoplasmas in 8 (10.1%) dogs. Thus, hemoplasma infections also showed a particularly high prevalence in this dog population. Based on a partial fragment of the 18S rRNA gene, B. gibsoni from Hungary exhibited complete sequence identity with conspecific strains reported from Europe and Asia. The cytochrome c oxidase subunit 1 (cox1) gene sequence of this isolate showed the closest identity with B. gibsoni reported from Japan but had a nonsynonymous mutation (M33I). Furthermore, the 11 B. gibsoni-positive samples analyzed for sequence variants of the cytochrome b (cytb) gene showed the presence of a common mutation (P310S). Most importantly, B. gibsoni had two further nonsynonymous mutations, M121I and F258L, in a dog with severe and relapsing anemia following atovaquone treatment. Phylogenetically, both cytb sequence variants clustered together, with a clear geographical pattern showing the closest relationship of both haplotypes identified in Hungary with those from China and Japan. To the best of our knowledge, this is the first cox1 and cytb characterization of B. gibsoni in Europe, as well as the first report on the emergence of this piroplasm and hemoplasmas with high prevalence among “fighting dogs” north of the Mediterranean Basin.

Abstract Background Anopheles stephensi, the major vector of urban malaria in India, was recently detected for the first time in Sri Lanka in Mannar Island on the northwestern coast. Since there are different biotypes of An. stephensi with different vector capacities in India, a study was undertaken to further characterise the genotype and biotype of An. stephensi in Mannar Island. Methods Mosquito larvae were collected in Pesalai village in Mannar and maintained in the insectary until adulthood. Adult An. stephensi were identified morphologically using published keys. Identified adult An. stephensi were molecularly characterized using two mitochondrial (cox1 and cytb) and one nuclear (ITS2) markers. Their PCR-amplified target fragments were sequenced and checked against available sequences in GenBank for phylogenetic analysis. The average spiracular and thoracic lengths and the spiracular index were determined to identify biotypes based on corresponding indices for Indian An. stephensi. Results All DNA sequences for the Mannar samples matched reported sequences for An. stephensi from the Middle East and India. However, a single nucleotide variation in the cox1 sequence suggested an amino acid change from valine to methionine in the cox1 protein in Sri Lankan An. stephensi. Morphological data was consistent with the presence of the Indian urban vector An. stephensi type-form in Sri Lanka. Conclusions The present study provides a more detailed molecular characterization of An. stephensi and suggests the presence of the type-form of the vector for the first time in Sri Lanka. The single mutation in the cox1 gene may be indicative of a founder effect causing the initial diversification of An. stephensi in Sri Lanka from the Indian form. The distribution of the potent urban vector An. stephensi type-form needs to be established by studies throughout the island as its spread adds to the challenge of maintaining the country’s malaria-free status.

Eleutherodactylus planirostris (Cope 1862), commonly called the Greenhouse Frog, is an insectivorous, direct-developing frog native to the Caribbean. It has been widely introduced outside of its native range and has been known to reach population densities of about 12,500 frogs per hectare, posing a potential ecological threat in areas of its introduction, especially to local insect populations. Recently, the species has been detected on several islands throughout the Philippines. Samples for this study were obtained from two locations in Quezon City (Luzon Island) and one location in Bacolod City (Negros Island). DNA barcoding using three genes (Cytochrome b, 16S rDNA, and Cytochrome Oxidase subunit 1) was performed with the objective of identifying the source population of Philippine E. planirostris. Our results indicate that E. planirostris samples in the Philippines are identical genetically to populations in Hawai'i and Florida, USA and are closely related to an individual from Matanzas, Cuba. A haplotype network built using the Fitch algorithm also supports the Cuban origin of the Philippine samples. Moreover, the Philippine specimens have nearly identical sequences for all three genes, which may have implications on its success as an introduced species. [ABSTRACT FROM AUTHOR]

Background: Anopheles stephensi, the major vector of urban malaria in India, was recently detected for the first time in Sri Lanka in Mannar Island on the northwestern coast. Since there are different biotypes of An. stephensi with different vector capacities in India, a study was undertaken to further characterise the genotype and biotype of An. stephensi in Mannar Island. Methods: Mosquito larvae were collected in Pesalai village in Mannar and maintained in the insectary until adulthood. Adult An. stephensi were identified morphologically using published keys. Identified adult An. stephensi were molecularly characterized using two mitochondrial (cox1 and cytb) and one nuclear (ITS2) markers. Their PCR-amplified target fragments were sequenced and checked against available sequences in GenBank for phylogenetic analysis. The average spiracular and thoracic lengths and the spiracular index were determined to identify biotypes based on corresponding indices for Indian An. stephensi. Results: All DNA sequences for the Mannar samples matched reported sequences for An. stephensi from the Middle East and India. However, a single nucleotide variation in the cox1 sequence suggested an amino acid change from valine to methionine in the cox1 protein in Sri Lankan An. stephensi. Morphological data was consistent with the presence of the Indian urban vector An. stephensi type-form in Sri Lanka. Conclusions: The present study provides a more detailed molecular characterization of An. stephensi and suggests the presence of the type-form of the vector for the first time in Sri Lanka. The single mutation in the cox1 genemay be indicative of a founder effect causing the initial diversification of An. stephensi in Sri Lanka from the Indian form. The distribution of the potent urban vector An. stephensi type-form needs to be established by studies throughout the island as its spread adds to the challenge of maintaining the country's malaria-free status. [ABSTRACT FROM AUTHOR]

Present study aimed to address molecular diversities of the small five-toed jerboa Allactaga elater Lichtenstein, 1825, and the Toussi jerboa Allactaga toussi Darvish et al., 2008, in marginal geographic distribution of the genus in Iran. The study involved 35 individuals of A. elater and A. toussi, from the east and central parts of Iranian plateau. The two probabilistic phylogenetic algorithms, Maximum Likelihood (ML) and Bayesian Inference (BI), applied on 68 sequences of the two mitochondrial genes (34 cytochrome b and 34 cytochrome c oxidase subunit 1), retrieved reciprocal monophyly of the two species. Independent species status of A. elater and A. toussi is further evident from their sympatry in eastern Iran. Each of these species was further subdivided into two deeply divergent phylogeographic lineages within Iranian plateau, showing high level of genetic divergence ranging from 7-10.7 % for cytb and 7.8-12.4 % for cox1 genes. Such values exceed the intraspecific level of variation in rodents. [ABSTRACT FROM AUTHOR]

Copyright of Acta Scientiarum: Biological Sciences is the property of Universidade Estadual de Maringa and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Acta Scientiarum: Biological Sciences is the property of Universidade Estadual de Maringa and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Sarcocysts were detected in routinely processed histological sections of skeletal muscle, but not cardiac muscle, of two adult male otters ( Lutra lutra; Mustelidae) from northern Norway following their post-mortem examination in 1999 and 2000. The sarcocysts were slender, spindle-shaped, up to 970 μm long and 35-70 μm in greatest diameter. The sarcocyst wall was thin (∼0.5 μm) and smooth with no visible protrusions. Portions of unfixed diaphragm of both animals were collected at the autopsies and kept frozen for about 14 years pending further examination. When the study was resumed in 2013, the thawed muscle samples were examined for sarcocysts under a stereo microscope, but none could be found. Genomic DNA was therefore extracted from a total of 36 small pieces of the diaphragm from both otters, and samples found to contain Sarcocystidae DNA were used selectively for PCR amplification and sequencing of the nuclear 18S and 28S ribosomal (r) RNA genes and internal transcribed spacer 1 (ITS1) region, as well as the mitochondrial cytochrome b ( cytb) and cytochrome c oxidase subunit 1 ( cox1) genes. Sequence comparisons revealed that both otters were infected by the same Sarcocystis sp. and that there was no genetic variation (100 % identity) among sequenced isolates at the 18S and 28S rRNA genes (six identical isolates at both loci) or at cox1 (13 identical isolates). PCR products comprising the ITS1 region, on the other hand, had to be cloned before sequencing due to intraspecific sequence variation. A total of 33 clones were sequenced, and the identities between them were 97.9-99.9 %. These sequences were most similar (93.7-96.0 % identity) to a sequence of Sarcocystis kalvikus from the wolverine in Canada, but the phylogenetic analyses placed all of them as a monophyletic sister group to S. kalvikus. Hence, they were considered to represent a novel species, which was named Sarcocystis lutrae. Sequence comparisons and phylogenetic analyses based on sequences of the 18S and 28S rRNA genes and cox1, for which little or no sequence data were available for S. kalvikus, revealed that S. lutrae otherwise was most closely related to various Sarcocystis spp. using birds or carnivores as intermediate hosts. The cox1 sequences of S. lutrae from the otters were identical to two sequences from an arctic fox, which in a previous study had been assigned to Sarcocystis arctica due to a high identity (99.4 %) with the latter species at this gene and a complete identity with S. arctica at three other loci when using the same DNA samples as templates for PCR reactions. Additional PCR amplifications and sequencing of cox1 (ten sequences) and the ITS1 region (four sequences) using four DNA samples from this fox as templates again generated cox1 sequences exclusively of S. lutrae, but ITS1 sequences of S. arctica, and thus confirmed that this arctic fox had acted as intermediate host for both S. arctica and S. lutrae. Based on the phylogenetic placement of S. lutrae, the geographical location of infected animals (otters, arctic fox) and the distribution of carnivores/raptors which may have interacted with them, the white-tailed eagle ( Haliaeetus albicilla) seems to be a possible definitive host of S. lutrae. Some of the muscle samples from both otters were shown to harbour stages of Toxoplasma gondii through PCR amplification and sequencing of the entire ITS1 region (five isolates) and/or the partial cytb (eight isolates) and cox1 (one isolate). These sequences were identical to several previous sequences of T. gondii in GenBank. Thus, both otters had a dual infection with S. lutrae and T. gondii. [ABSTRACT FROM AUTHOR]

The five-toed jerboas of the genus Allactaga inhabit arid areas of Asia and North Africa and are widely distributed in deserts, semideserts, and steppe regions of Iran. Many morphological and morphometric studies have been carried out on this genus; however, there is little molecular data available on them. Therefore, their phylogenetic relationships need to be evaluated. Here, using sequences of 903 and 632 bp of two mitochondrial DNA genes, cytochrome b ( Cytb) and cytochrome oxidase subunit I ( Cox1), the phylogenetic relationships of five nominal species of Allactaga were evaluated from the Iranian Plateau. Phylogenetic trees were constructed on the basis of maximum parsimony, maximum likelihood, and Bayesian analyses. The resulting trees supported seven major clades: (1) Allactaga firouzi and A. hotsoni, (2) A. williamsi, (3) A. euphratica, (4) A. elater from Esfahan and Tehran, (5) A. toussi, (6) A. elater from Golestan, and (7) A. elater from Kashmar and Golestan. On the basis of the results of current study, A. firouzi and A. hotsoni are synonymized, while A. elater could be considered a complex species with three subclades. In addition, the geographic distribution analysis of the species was clarified and as a result, A. euphratica is a new record for the Ilam region. [ABSTRACT FROM AUTHOR]

Metacercariae of trematodes from the genus Diplostomum are major helminth pathogens of freshwater fish, infecting the eye or the brain. The taxonomy of the genus Diplostomum is complicated, and has recently been based mainly on the molecular markers. In this study, we report the results of the morphological and molecular genetic analysis of diplostomid metacercaria from the brain of the minnow Phoxinus phoxinus from three populations in Fennoscandia (Northern Europe) and one population in Mongolia (East Asia). We obtained the data on the polymorphism of the partial mitochondrial cox1 gene and ribosomal ITS1–5.8S–ITS2 region of these parasites. РСА-based morphological analysis revealed that the parasites in the Asian and the European groups of Diplostomum sp. were distinctly different. Metacercariae from the brain of Mongolian minnows were much larger than those from the brain of Fennoscandian minnows but had much fewer excretory granules. Considering that the two study regions were separated by a distance of about 4500 km, we also tested the genetic homogeneity of their host, the minnow, using the mitochondrial cytb gene. It was shown that Diplostomum -infected minnows from Mongolia and Fennoscandia represented two previously unknown separate phylogenetic lineages of the genus Phoxinus. Both molecular and morphological analysis demonstrated that the parasites from Fennoscandia belonged the species Diplostomum phoxini , while the parasites from Mongolia belonged to a separate species, Diplostomum sp. MТ. Each of the two studied Diplostomum spp. was associated with a specific, and previously unknown, genealogical lineage of its second intermediate host, P. phoxinus. • Two distinct species of ' Phoxinus phoxinus ' complex inhabit in Fennoscandia and Mongolia. • Two distinct species of ' Diplostomum phoxini ' complex parasitize brain of minnows in Fennoscandia and Mongolia. • Each of brain species is associated with specific genealogical lineages (European and Asian) of minnow as the second intermediate fish host. [ABSTRACT FROM AUTHOR]

Monoxenous parasites may easily infect animals in captivity, and nematodes belonging to the genus Trichuris are commonly reported in zoological gardens worldwide. Infections in captive animals should be accurately monitored and the characterization of pathogens is highly advisable, as a tool to infer possible routes of intra- and interspecific transmission pathways and to assess the related zoonotic potential. Whipworms are usually identified on the basis of few morphological features of adults males and eggs and by an host-affiliation criterion. Given the strong morphological convergence of adaptive traits and the possible occurrence of hybridization and/or cross-infections events, the use of molecular methods is of great utility. Here, we analysed two partial mitochondrial loci, the cytochrome c oxidase I and the cytochrome b regions, in Trichuris spp. infecting four animal species hosted in the Bioparco Zoological Garden of Rome. Results from molecular systematics, compared to previous data, suggested that the five Trichuris taxa recovered were well separated, showing a significant degree of host affiliation (herbivorous, primates/swine and rodents/canids). The screw horn antelopes and the camels were infected with two variants of Trichuris ovis ; Trichuris sp. similar to those observed in rodents from South America was infecting the Patagonian maras. Moreover, Trichuris from the ring-tailed lemur showed a great similarity to Trichuris infecting the Japanese macaque previously analysed from the same zoological garden, and clustering together with Trichuris trichiura , posing a potential zoonotic threat for visitors and workers. Unlabelled Image • Different Trichuris spp. infected four animal species hosted in the Bioparco Zoological Garden of Rome. • Trichuris sp. similar to whipworms found in south-american rodents infected the Patagonian mara. • Two variant of Trichuris ovis infected the screw horn antelope and the camel. • Two taxa related to T. trichiura infected the ring-tailed lemur suggesting potential zoonotic risk. [ABSTRACT FROM AUTHOR]