Partial Mitochondrial Gene Research Articles

A new species of Neoergasilus Yin 1956 (Copepoda: Cyclopoida: Ergasilidae) parasitic on the catfish Clarias gariepinus (Burchell, 1822) (Siluriformes: Clariidae) from South Africa

Twenty-one specimens of an ergasilid were collected from the gills of the sharptooth catfish, Clarias gariepinus (Burchell), from the Eastern Cape, South Africa. The first leg and antennae morphology conformed to the genus Neoergasilus Yin, 1956. While the invasive Neoergasilus japonicus (Harada, 1930) is the only Neoergasilus species that has been reported from Africa, the combination of several characteristics, separates the Eastern Cape specimens from N. japonicus and the other eight Neoergasilus congeners. These include an inflated cephalothorax; the presence of an oval dorsal ornamentation anterior to the cephalosome; a spine on the posterodistal margin of the first antennal segment; a cone-like process at the proximal margin of the second antennal segment; a knob-like process on the inner distal margin of the first exopodal segment of leg 1 and two forked spines on the third exopodal segment; leg 4 bearing a 2-segmented exopod and 3-segmented endopod; a single-segmented fifth leg with a seta extending from the base of the pedigerous somite and three unequal setae on its free segment; and a median caudal rami seta with an array of spines. Supporting genetic data were generated using two partial ribosomal RNA genes, 18S and 28S, and one partial mitochondrial DNA gene, COI. The Eastern Cape species is here proposed as new to science and described as Neoergasilus africanusn. sp. (Ergasilidae: Cyclopoida). This is the first Neoergasilus species described from the sharptooth catfish and from the southern hemisphere. Additionally, a key to all the species of this genus is provided.

Uncovering further diversity of Ochoterenatrema Caballero, 1943 (Digenea: Lecithodendriidae) in South American bats

Ochoterenatrema Caballero, 1943 is a genus of lecithodendriid digeneans that prior to this study included 8 species parasitic in bats in the Western Hemisphere. Species of Ochoterenatrema possess a unique morphological feature in form of the pseudogonotyl on the sinistral side of the ventral sucker. In this study, we describe 2 new species of Ochoterenatrema from bats in Ecuador. The new species are readily differentiated from their congeners by a combination of morphological characters, including the distribution of vitelline follicles, length of oesophagus, sucker ratio and the body shape, among other features. We have generated partial nuclear 28S rDNA and mitochondrial cox1 gene DNA sequences from both new species. The newly obtained sequences were used to differentiate among species and study the phylogenetic interrelationships among Ochoterenatrema spp. The internal topology of the clade was weakly supported, although the cox1 tree was much better resolved than the 28S tree. Comparison of sequences revealed 0–1.2% interspecific divergence in 28S and 3.3–20.5% interspecific divergence in cox1 among Ochoterenatrema spp. The new findings demonstrate that bats in South America likely harbor multiple additional undescribed species of Ochoterenatrema. More extensive sampling from broader geographic and host ranges, especially in North America, should allow for a better understanding of the evolution of host associations and morphological traits of this lineage of lecithodendriid digeneans.

Mitogenomic Analysis and Phylogenetic Implications for the Deltocephaline Tribe Chiasmini (Hemiptera: Cicadellidae: Deltocephalinae).

The grassland leafhopper tribe Chiasmini (Cicadellidae: Deltocephalinae) presently comprises 324 described species worldwide, with the highest species diversity occurring in the Nearctic region but a greater diversity of genera occurring in the Old World. In China, this tribe comprises 39 described species in 11 genera, but the fauna remains understudied. The complete mitogenomes of three species of this tribe have been sequenced previously. In order to better understand the phylogenetic position of Chiasmini within the subfamily Deltocephalinae and to investigate relationships among Chiasmini genera and species, we sequenced and analyzed the complete mitogenomes of 13 species belonging to seven genera from China. Comparison of the newly sequenced mitogenomes reveals a closed circular double-stranded structure containing 37 genes with a total length of 14,805 to 16,269 bp and a variable number of non-coding A + T-rich regions. The gene size, gene order, gene arrangement, base composition, codon usage, and secondary structure of tRNAs of the newly sequenced mitogenomes of these 13 species are highly conserved in Chiasmini. The ATN codon is commonly used as the start codon in protein-coding genes (PCGs), except for ND5 in Doratura sp. and ATP6 in Nephotettix nigropictus, which use the rare GTG start codon. Most protein-coding genes have TAA or TAG as the stop codon, but some genes have an incomplete T stop codon. Except for the tRNA for serine (trnS1(AGN)), the secondary structure of the other 21 tRNAs is a typical cloverleaf structure. In addition to the primary type of G-U mismatch, five other types of tRNA mismatches were observed: A-A, A-C, A-G, U-C, and U-U. Chiasmini mitochondrial genomes exhibit gene overlaps with three relatively stable regions: the overlapping sequence between trnW and trnC is AAGTCTTA, the overlapping sequence between ATP8 and ATP6 is generally ATGATTA, and the overlapping sequence between ND4 and ND4L is generally TTATCAT. The largest non-coding region is the control region, which exhibits significant length and compositional variation among species. Some Chiasmini have tandem repeat structures within their control regions. Unlike some other deltocephaline leafhoppers, the sequenced Chiasmini lack mitochondrial gene rearrangements. Phylogenetic analyses of different combinations of protein-coding and ribosomal genes using maximum likelihood and Bayesian methods under different models, using either amino acid or nucleotide sequences, are generally consistent and also agree with results of prior analyses of nuclear and partial mitochondrial gene sequence data, indicating that complete mitochondrial genomes are phylogenetically informative at different levels of divergence within Chiasmini and among leafhoppers in general. Apart from Athysanini and Opsiini, most of the deltocephaline tribes are recovered as monophyletic. The results of ML and BI analyses show that Chiasmini is a monophyletic group with seven monophyletic genera arranged as follows: ((Zahniserius + (Gurawa + (Doratura + Aconurella))) + (Leofa + (Exitianus + Nephotettix))).

Molecular phylogenetic analysis of Echinococcus granulosus sensu lato infecting sheep in Italy.

Cystic Echinococcosis (CE), caused by the larval form of Echinococcus granulosus sensu lato, is a neglected zoonosis still threatening public health worldwide. In Italy different epidemiological scenarios were reported depending on the geographical area and associated socio-economic activities. Although in northern Italy the occurrence of E. granulosus is considered sporadic, in the southern regions and, particularly in Sardinia, CE prevalence reaches high levels. We analysed CE cysts collected from infected sheep from various areas of mainland Italy and the Sardinia island, with the main objective to investigate intergenotypic and intragenotypic variations at national level. CE cysts were collected from slaughtered sheep following post mortem inspection at local abattoirs. Total genomic DNA was extracted and amplification and sequencing of the partial mitochondrial genes nad5 and cox1 were performed. A Bayesian phylogenetic tree was estimated on a nad5 dataset (n=260) composed of E. granulosus samples from this study (n=126) and all the nad5 haplotypes available in GenBank (n=134). In addition, haplotype network, diversity and neutrality analysis were performed on nad5 and cox1 sequences of Italian origin obtained in this study. E. granulosus sensu stricto (s.s.) was found to be the only Echinococcus species infecting sheep in Italy, mainly represented by G1 genotype (76%) and, to a lower extent, by G3 genotype (24%). Phylogenetic analyses revealed 40 nad5 and 33 cox 1 haplotypes, and the presence of two founder haplotypes, belonging to G1 and G3 genotype, showing 100% similarity with DNA sequences from different geographic regions. The lack of geographical segregation, high haplotype and low nucleotide diversity coupled with significant negative values of Tajima's D and Fu's Fs observed in this study indicated high genetic variation and demographic expansion of E. granulosus s.s. in Italy.

Exploring the Potential in Combining the Two Molecular Approaches, COI Barcoding and PCR-RFLP (COIBar-RFLP) in Identification of Selected Species of the Family Carangidae

The family Carangidae is considered a markedly diverse, widespread taxon. Due to the characteristic “cryptic diversity” and “hybrid speciation” within the family, there is an exigency for taxonomic approaches that go beyond traditional phenotypic modus in identifying species. Mitochondrial cytochrome c oxidase subunit - I (COI) barcoding gene region plays a significant role over phenotypic characters in identifying species. This study evaluates the combination of the two molecular approaches: the COI DNA barcoding and PCR-RFLP for judicious species discrimination. The partial mitochondrial COI gene region of the selected Carangid fish species was amplified, sequenced and confirmed sequences with a mean length of 655 bp were submitted to the main databases; NCBI and BOLD. Intraspecific and interspecific nucleotide divergences were computed by the Kimura-2-Parameter (K2P) and they ranged between (0.00–2.96 %) and (6.46–21.83 %), respectively. The possibility of acquiring the same RFLP profiles given by the restriction enzymes HaeIII and MbOII was observed by the theoretical cleavage of 250 reference sequences of the corresponding gene. Hence, major and minority composite haplotypes of each species were obtained based on the fragment types derived by both HaeIII and MbOII. The resulting divergence values were compatible with the previously reported values for marine fish species. All the species were clearly differentiated by both RFLP banding patterns and the highest probability assumption of getting the same RFLP profile was compatible with the most abundant composite haplotype of each species. This reveals the practicability of the combination of two consolidated molecular approaches, COI barcoding and PCR-RFLP (COIBar-RFLP).

Confirmation of the valid specific status of Dolichovespulakuami Kim & Yoon, 1996 (Hymenoptera, Vespidae) based on molecular and morphological evidence.

The taxonomic validity of Dolichovespulakuami, especially in relation to D.flora, has been the subject of a long-term debate. Herein, the valid specific status of the former was supported through an integrated analysis of morphological characters and DNA barcodes. The pronotal rugae and male genitalia of the two species are different, and partial mitochondrial genes (cytochrome oxidase subunit I, COI) indicate that they form significantly distinct lineages. The hitherto unknown male of D.kuami is described for the first time, and a brief discussion of the D.maculata species group is provided.

A phylogenetic study of the Mongolian Tree Pipit (Anthus trivialis, Linnaeus, 1758) population based on mitochondrial DNA (mtDNA) genes

Our aim was to identify nucleotide polymorphisms, assess their distribution in haplotype diversity, and construct a phylogenetic tree by analyzing mtDNA markers of the Mongolian Tree Pipits(Anthus trivialis). We conducted this study using partial gene sequencesof mitochondrial marker genes, such as COI, Cyt-b, D-loop, andND2,to determine the genetic diversity of Mongolian Tree Pipits. We successfully amplified 2307 bp of the mitochondrial DNA fragments, including 469bp of COI, 435bp of Cyt-b, 554bp of D-loop and 846bp of ND2 from total 27 individuals of Mongolian (21) and Hungarian (6) populations of Anthus trivialis.The Hd value wasthe highest for ND2 (0.96) as compared with the other gene fragments in all populations, whileit was 0.94 in the Mongolian population. Moreover, the nucleotide diversity (Pi) ranged from 0.00234 to 0.004 in all population, it was observed that the Pi was between 0.00183 and 0.00376 in the Mongolian population. The phylogenetic tree based on combined mtDNA sequences revealed two mean clades. The probability value of the node supporting the posterior between these clades is 0.65,which suggests an indicative support in relationship between the two clades. Furthermore, phylogenetic analysis showed that Mongolian Tree Pipits do share common genetic characteristics with other populations and do not form distinct clusters. Publisher correction: Author name Sabor Sramko was revised to Gabor Sramko dated on 2024-10-07.

First mitochondrial genome of Amblyomma triste Koch, 1844 (Acari: Ixodidae): Evidence for studying species within the A. maculatum group</p >

The tick species that comprise the Amblyomma maculatum group (Acari: Ixodidae) are widely distributed in the Nearctic and Neotropical regions, ranging from the United States of America (USA) to Argentina. This group includes three species: Amblyomma maculatum, Amblyomma tigrinum, and Amblyomma triste, which parasitize a high number of vertebrates, including domestic mammals and humans, where they are the main vectors of Rickettsia parkeri s.s. The identification, distribution, and validity of the three species within the group have been controversial and continue to raise questions. Previous studies have mostly focused on the analysis of partial nuclear and mitochondrial genes, highlighting the need for new integrative analyses that contribute to clarifying their systematics and ecology. In this investigation, we obtained the first mitochondrial genome of A. triste (14,808 bp), which, upon comparison with the reported mitochondrial genomes of A. maculatum, exhibits a genetic distance of 4.2%, providing new evidence for the validity of the former species. The order, composition, and structure of the mitogenome of A. triste are consistent with the characteristics reported for most Metastriata including taxa within Amblyomma, that count with several mitochondrial genomes published in the last few years. The attributes of the mitogenomes, such as the absence of paralogs and a small genome, confirm the utility in studies involving complexes of cryptic species as it also serves as a robust tool for inferring phylogenies. Furthermore, we reported new molecular markers at the mitochondrial level which can be used in phylogenetic studies for other tick species, especially those with controversial or challenging taxonomy. We highlight the need of sequencing the mitochondrial genome of A. tigrinum, which is part of the A. maculatum group, as well as experimental crosses of populations from different places in the Americas to obtain additional evidence for species recognition.

Examination of sequence variations in partial mitochondrial 12S gene amongst damselfish species as references for DNA barcoding.

Accurate species identification, based on DNA barcoding, can be achieved when sufficient sequence variations are present amongst species in the sampled marker. In general, the ability to discriminate species decreases with shorter sequences; however, shorter regions have a merit in amplification success by the polymerase chain reaction. In either case, it is important to investigate sequence variations amongst species before barcoding to understand its reliability and limitations. In this study, we investigate how accurately short, but hypervariable portion of the mitochondrial 12S ribosomal RNA (12S) gene (MiFish region with approximately 180 bp) is used to identify each species in diversified pomacentrid fishes compared with the longer region of the same gene (approximately 750 bp). We prepared three datasets with 301 sequences of the MiFish region for 150 species, the same 301 of sequences of the longer 12S region and 476 sequences of the MiFish region for 183 species. Neighbour-joining (NJ) analyses and genetic distance analyses revealed several indistinguishable pairs of species in these DNA regions. Although the number of such pairs was larger in the MiFish region, 83.6% (153 of 183) of species possessed respective unique sequences even in the MiFish region (versus 96.0% [144 of 150 species] in the longer 12S region). A part of indistinguishable pairs of species might have caused by mitochondrial DNA introgressions and taxonomically unresolved problems. Our analysis clarified the effectiveness and limitations of species identification using DNA barcoding for Pomacentridae and the sequences we provided here contribute to the expansion of references for pomacentrid mitochondrial 12S sequences.

New records of Ripersiella (Tinsley) (Hemiptera: Coccomorpha: Rhizoecidae) species and identification key of Rhizoecidae in Italy

Biological invasions are one of the major constraints worldwide because of the economic and environmental consequences they may pose. Root mealybugs (Hemiptera: Coccomorpha: Rhizoecidae) are continuously transported worldwide with plant soil because of their cryptic behavior. In this study it was reported the presence of Ripersiella multiporifera Jansen in Sicily for the first time and discussed the presence of R. maasbachi (Jansen) and R. hibisci (Kawai & Takagi) in Italy. A key is provided to identify the species of Rhizoecidae currently known in Italy. The species were identified by morphological features and characterized molecularly by using a partial COI mitochondrial gene. Our report alerts the presence of alien root mealybugs in Italy. R. hibisci is a quarantine species listed in Annex IIA of EU Regulation 2019/2072.

Exploring gene sequences and phylogenetic relationships of four terrestrial planarian species (Platyhelminthes; Tricladida; Geoplanidae) in Europe.

Partial mitochondrial cox1 gene sequences from four recently recognised European species of terrestrial planarians, and ribosomal ITS1 sequences for two of them, are presented: Marionfyfea adventor, Artioposthia exulans (both introduced from New Zealand), Australopacifica atrata (from Australia) and specimens putatively identified as Microplana edwardsi, presumed to be native to the UK. The sequences are compared with those from other terrestrial planarian species and analysed phylogenetically. Results indicate that the sister group of M. adventor comprises a clade constituted by at least the genus Arthurdendyus. The phylogenetic position of Ar. exulans remains less certain, Australopacifica atrata might be closely related to the species Parakontikia ventrolineata and Endeavouria septemlineata. The specimens of M. cf. edwardsi are distinct from all other Microplana species for which sequences are available.

A new snake of the genus Dendrelaphis Boulenger, 1890 (Squamata: Colubridae) from the coastal area of southern Vietnam.

We describe a new species of Dendrelaphis from the coastal area in southern Vietnam based on morphological data and nucleotide sequences from one partial mitochondrial gene (Cytb). Dendrelaphis binhi sp. nov. is diagnosed by the following morphological characters: body scale rows 13 at neck and midbody, 9-11 before vent; vertebral scale row feebly enlarged; ventrals 154-158 in males and 161-170 in females; subcaudals 95-106 in males and 95-102 in females; one supralabial touching the eye; 18 or 19 maxillary teeth; hemipenis spinose, reaching 10th or 11th SC, with a slender papilla; cloacal plate divided; dorsum dark brown; and a white stripe along the lower flank. The new species differs from its congeners by an uncorrected p-distance in Cytb sequences of at least 8.1%.

Genetic Variations of Malaysian And Golden Thai Strains of Climbing Perch and Their Hybrids Based On The Partial Mitochondrial Cytochrome Oxidase Subunit 1 Gene

Rapid genetic improvements from selective breeding are anticipated in many aquaculture species and mitogenome is used to complement the morphological taxonomy of hybrids to evaluate its genetic structures. A study of the genetic variations within the two strains (Malaysian strain and Golden Thai strain) of Anabas testudineus (Bloch, 1792) and their hybrids, phylogenetic trees based on cytochrome oxidase subunit 1 (CO1) of the partial mitochondrial DNA gene were constructed using Maximum Likelihood and Neighbor Joining approaches. The findings support the monophyletic status of the genus with only one haplogroup from which other haplotypes were descended and of a single common ancestor. The individual fishes’ phylogenetic relationships revealed two major clades and Saurida undosquamis as an outgroup. All the groups had high haplotype diversity, except for the hybrid Malaysian strain × golden Thai strain (0.1540±0.126). This suggests that different fish species in each of these studies had different nucleotide compositions in their mitochondrial genomes. The highest number of haplotypes and the presence of distinct haplotypes in the Golden Thai strain × Malaysian strain hybrid point to the absence of recent or regular gene flow as well as high genetic diversity within the hybrids. This study demonstrated that the mtDNA diversity of Anabas testudineus from Malaysia and Thailand had been preserved. Studies on the population genetic diversity and molecular evolution of anabas fish species can benefit from the data provided by this work.

Nasopharyngeal myiasis due to Cephalopina titillator in Southeastern Iran: a prevalence, histopathological, and molecular assessment.

The Cephalopina titillator is one of the most important causative agents of nasal myiasis in camels. This study aimed to explore the prevalence, histopathological effects, and molecular identification of C. titillator infestation in camels of Kerman province, South-Eastern Iran, between 2019 and 2021. The larvae were placed in 10% formalin for histopathological evaluation and species identification. Pieces of larval abdominal segments of C. titillator were selected for extraction of DNA. Partial mitochondrial CO1 genes were sequenced for final analysis. Out of the 870 camels examined, 339 (38.9%) were infested with larval stages of C. titillator. There was a significant difference between age and infection rate (P = 0.001), while no association between males and females (P = 0.074) was found. The infection rate was significantly higher in the winter (P < 0.001) than in the other seasons. In this study, different lesions depending on duration, locations, and the depth of larval adhesion notably degeneration changes, necrosis, and ulceration were observed. Also, in chronic cases, granulation tissue reactions were organized. Cephalopina titillator was confirmed by PCR sequencing analysis using mitochondrial CO1 region. A 582bp nucleotide sequence was deposited in GenBank under the MW136151 accession number. Phylogenetic analysis of CO1 produced a single uniform sister clade to MZ209004 and MW167083 records from China and Iraq, respectively. The high prevalence of C. titillator in camels in this region and other areas of Iran declares that the country is in an endemic status and displays the existence of the potential risk for camels.

Two new deep-water species of squat lobsters (Crustacea: Anomura: Galatheoidea) from the Central and Southwest Indian Ridge

The high diversity and endemism of benthic species on seamounts are extensively documented. Although the number of squat lobster species from seamount habitats has increased, species from the Central Indian Ridge (CIR) and Southwest Indian Ridge (SWIR) systems remain largely unknown. The hydrothermally active Indian Ridge system harbours potential sites for future Seafloor Massive Sulphides (SMS) mining. Since seabed mining may create a long-lasting impact on the deep-sea ecosystems, it could harm the unique deep-sea life associated with mineral deposits. This study identified two undescribed squat lobster specimens: one from the genus Munidopsis Whiteaves, 1874 from the CIR system in water depths ranging from 1981 to 2033 m, and one from the genus Typhlonida Macpherson & Baba in Machordom, Ahyong, Andreakis, Baba, Buckley, Garcia-Jimenez, McCallum, Rodriguez-Flores & Macpherson, 2022 from the SWIR system in depths ranging from 2070 to 2404 m. Both new species may be distinguished morphologically and genetically based on the partial mitochondrial (mtCOI) gene. To generate baseline environmental and biodiversity data, efforts for accurate sampling and correct identification of biological species are required.

Phylogenetic relationships of the Amblyomma cajennense complex (Acari: Ixodidae) at mitogenomic resolution

The genus Amblyomma is the third most diverse in the number of species within the Ixodidae, with practically half of its species distributed in the Americas, though there are also species occurring in Africa, Asia, and Australia. Within the genus, there are several species complexes with veterinary and public health importance. The Amblyomma cajennense complex, in the Americas, is represented by six species with a wide distribution, from Texas to northern Argentina. We combined two sequencing techniques to generate complete mitogenomes of species belonging to the Amblyomma cajennense complex: genome skimming and long-range PCRs sequencing methods. Thus, we generated seven new mitochondrial genomes for all species of the Amblyomma cajennense complex, except for Amblyomma interandinum. Genetic distances between the mitogenomes corroborate the clear differentiation between the five species of the Amblyomma cajennense complex. The phylogenetic relationships of these species had previously been evaluated by combining partial nuclear and mitochondrial genes and here these relationships are corroborated with a more robust framework of data, which demonstrates that the conjunction of mitochondrial and nuclear partial genes can resolve close relationships when entire genes or genomes are unavailable. The gene order, structure, composition, and length are stable across these mitogenomes, and they share the general characteristics of Metastriata. Future studies should increase the number of available mitogenomes for this genus, especially for those species from the Indo-Pacific region and Africa, by means of a better understanding of their relationships and evolutionary process.

Diversity of helminths with zoonotic potential and molecular characterization of Toxocara canis infecting domestic dogs from locations of Amazon and Atlantic Forest Brazilian biomes.

The coproparasitological examination of dogs (n=278) from two Brazilian biomes (Amazon [AZ] and Atlantic Forest [AF]) by centrifugal flotation demonstrated positivity values of 54.2% (AF) and 48.5% (AZ). The most prevalent parasites in AF were hookworms (81.0% - 47/58), Toxocara sp. (17.3% - 10/58) and Trichuris vulpis (12.1% - 7/58); while in AZ they were hookworms (86.7% - 72/83), Toxocara sp. (18.1% - 15/83), Dipylidium caninum (13.3% - 11/83) and T. vulpis (10.8% - 9/83). PCR was performed using the partial mitochondrial genes cytochrome c oxidase subunit 1 (pcox1) and NADH dehydrogenase 1 (pnad1) in 25 fecal samples positive for Toxocara sp. eggs and found one sample positive for pcox1 and six positives for pnad1. The sequencing of these samples was unsuccessful due to the difficulties inherent in copro-PCR+sequencing. The sequencing of 14 samples of T. canis adult helminths retrieved 11 sequences of 414 bp for pcox1 and nine sequences of 358 bp for pnad1. The phylogenetic trees of these sequences confirmed the species T. canis. Intraspecific genetic variation was only observed for pnad1. This is the second study involving molecular analysis of T. canis in dogs from Brazil and adds new information through the use of pnad1.

Description and Phylogenetic Relationships of Anhingatrema n. gen. (Digenea: Diplostomidae) with Two New Species from New World Anhingas (Aves: Anhingidae).

The Diplostomidae is a globally distributed family of digeneans that parasitize a wide variety of tetrapod definitive hosts. Recent molecular phylogenetic studies have revealed unknown diplostomid diversity in avian hosts throughout the New World. Herein, we provide descriptions of a novel genus of diplostomids with two new species. Two species of diplostomids belonging to the new genus were collected from anhinga birds in Mississippi (USA) and Brazil. Partial nuclear 28S ribosomal and mitochondrial cox1 genes were sequenced. Ribosomal data were used for phylogenetic inference. Both species of Anhingatrema n. gen. were positioned in a 100% supported, monophyletic clade in the phylogenetic tree. The molecular phylogenetic position and a combination of morphological features (e.g., presence of pseudosuckers, testes shape and orientation) supported erection of the new genus. Anhingatrema overstreeti n. sp. and Anhingatrema cararai n. sp. are morphologically similar, but differ in size of and ratios associated with pseudosuckers. The two species differ by 2% of 28S sequences and 13.8% of cox1 sequences. Comparison of DNA sequences revealed that Diplostomidae gen. sp. in GenBank (MZ314151) is conspecific with An. overstreeti n. sp. Anhingatrema n. gen. is the sixth genus of diplostomids known from anhingas worldwide. Anhingatrema cararai n. sp. is the first diplostomid to be reported from anhingas in South America. Combined with previous studies, the molecular phylogenies revealed at least two host switches to anhingas from other birds during the evolutionary history of the Diplostomidae.

The complete mitochondrial genome of Tuta absoluta (Lepidoptera: Gelechiidae) and genetic variation in two newly invaded populations in China

• The size of mitogenome sequences in T. absoluta varied among individuals from different geographical locations. • The differences in the lengths of intergenic spacer were contributed to the size difference of mitogenomes among different populations. • cox2 , atp6 , nad1 and nad5 genes were sensitive mitochondrial gene markers for population genetics in T. absoluta . Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) is a devastating invasive pest worldwide, causing severe damage to tomatoes. Recently, it has been recorded in the northwestern and southwestern parts of China. Here, the mitogenomes and genetic variation of two newly invaded T. absoluta populations in Xinjiang and Yunnan, were determined. The results showed that the complete mitogenome size of T. absoluta is 15298 bp for the individual from Xinjiang and 15296 bp for the individual from Yunnan, which were both longer than the reported mitogenome from Spain (15290 bp). The mitogenome sequences of individuals collected from three locations showed high levels of sequence similarity, except for 8 polymorphic sites, which were in genes cox2 (1 site), cox3 (2 sites), cob (1 site) , atp6 (1 site), nad1 (2 sites) and nad5 (1 site). Tuta absoluta mitogenomes share many features with other 6 Gelechiidae mitogenomes, except for several differences in the start and stop codons of protein-coding genes and the length of intergenic spacers. Seven partial mitochondrial genes ( cox1 , cox2 , cox3 , atp6 , cob , nad1 , and nad5 ) were used for genetic variation analysis, and significant population differentiation was found between the two populations based on cox2 , atp6 , nad1 , and nad5 . The complete mitogenomes and sensitive mitochondrial gene markers reported here provide useful data for further population genetics study of this pest.

A Novel PCR-RFLP to Detect and Differentiate Schistosoma spindale and S. indicum, the Pathogenic Schistosomes in Indian Cattle.

Visceral schistosomosis is an economically important trematode infection caused by Schistosoma spindale and S. indicum in among ruminants. The lack of sensitive diagnostic tools has often led to underestimation of the prevalence in live animals. A sensitive copro-PCR targeting partial mitochondrial gene was developed to detect Schistosoma spp. However, this protocol could not differentiate between the two species. This study was conducted to explore the possibility of species differentiation using restriction fragment length polymorphism of PCR products (PCR- RFLP). Polymerase chain reaction was carried out to amplify mitochondrial gene of adult S. spindale and S. indicum. Copro PCR was done with schistosome-positive faecal samples. A novel PCR-RFLP was designed targeting the Hpy166II recognition sequence in the mitochondrial gene sequence of S. indicum. The PCR using primers targeting the mitochondrial gene of S. spindale and S. indicum amplified a distinct product of approximately 454bp with adult fluke as well as faecal DNA, which upon RFLP with Hpy166II yielded 330bp and 124bp products with S. indicum amplicons alone. The novel PCR-RFLP possesses the potential to be used in epidemiological surveys among bovines and in snail intermediate hosts to screen for S. spindale and S. indicum infection.