The Impact of Artificial Barriers on the Varuna litterata Migration Route in the Lower Serayu River, Central Java and Its Molecular Identification

BackgroundAccurate identification of mosquito species is essential for the development and optimization of strategies to control mosquitoes and mosquito-borne diseases. Problems with the morphological identification of mosquito species have led to the use of molecular identification techniques, in particular the Folmer cytochrome c oxidase subunit I (COI) PCR system (FCOS), originally designed to identify a range of other invertebrates.MethodsAs there can be difficulties identifying mosquitoes using FCOS, we re-evaluated the FCOS primers and developed a new COI-based SYBR PCR (the Auburn COI system—AUCOS) to improve the molecular identification of mosquitoes. Sequence data in GenBank for 33 species from 10 genera of mosquitoes were used to develop our AUCOS primers. Two molecular assays (AUCOS, FCOS) and morphological identification were carried out on mosquitoes collected from the field in Auburn, Alabama (USA) and on Saint Kitts.ResultsWith a convenience sample of individual mosquitoes comprising 19 species from six genera in Saint Kitts (n = 77) and Auburn (n = 48), our AUCOS provided higher-quality sequence data than FCOS. It also proved more sensitive than FCOS, successfully amplifying 67.5% (85/126) as opposed to 16.7% (21/126) of the samples. The species determined by morphology, or genus with damaged samples, matched that as determined by AUCOS for 84.9% (62/73) of the samples. Morphological classification was confirmed by FCOS with 81.0% (17/21) of samples producing utilizable sequences. While both FCOS and AUCOS correctly identified all the Aedes, Anopheles, Deinocerites, and Uranotaenia species in the study, identification of Culex species was less successful with both methods: 50.0% (3/6) by FCOS and 35.7% (5/14) by AUCOS.ConclusionsThe AUCOS DNA barcoding system for mosquito species described in this study is superior to the existing FCOS for the identification of mosquito species. As AUCOS and FCOS amplify the same variable region of the COI, the large amount of existing data on GenBank can be used to identify mosquito species with sequences produced by either PCR.Graphical

Dear Editor, Multiple infections with small liver flukes and minute intestinal flukes are the serious public health concern in the lower Mekong basin [1,2]. Although the epidemiological survey for those trematode infections are primarily carried out based on copro-parasitological examination, detection/identification of fecal eggs/worms is a tedious job and often problematic because of the morphological similarities of eggs/worms. Along with the popularization of PCR-sequencing methods, copro-DNA diagnosis and molecular phylogenetic identification/speciation have been introduced in epidemiological studies. Among various genes and non-coding lesions of nuclear and mitochondrial DNAs, mitochondrial cytochrome c oxidase subunit I (COXI) is one of the most widely used inter- and intra-species marker. Using COXI and some other markers, Lee and his colleagues performed molecular phylogenetic analyses on small liver flukes (Lee SU, Huh S. Variation of nuclear and mitochondrial DNAs in Korean and Chinese isolates of Clonorchis sinensis. Korean J Parasitol 2004; 42: 145-148) and on minute intestinal flukes (Lee SU, Huh S, Sohn WM, Chai JY. Sequence comparisons of 28S ribosomal DNA and mitochondrial cytochrome c oxidase subunit I of Metagonimus yokogawai, M. takahashii and M. miyatai. Korean J Parasitol 2004; 42: 129-135). The COX1 gene sequences appeared in those articles are; Clonorchis sinensis ({type:entrez-nucleotide,attrs:{text:AF184619,term_id:296940320,term_text:AF184619}}AF184619, {type:entrez-nucleotide,attrs:{text:AF181889,term_id:285026682,term_text:AF181889}}AF181889, {type:entrez-nucleotide,attrs:{text:AF188122,term_id:285809842,term_text:AF188122}}AF188122), Metagonimus yokogawai ({type:entrez-nucleotide,attrs:{text:AF096230,term_id:297039733,term_text:AF096230}}AF096230), Metagonimus takahashii ({type:entrez-nucleotide,attrs:{text:AF096231,term_id:297039734,term_text:AF096231}}AF096231), Metagonimus miyatai ({type:entrez-nucleotide,attrs:{text:AF096232,term_id:297039735,term_text:AF096232}}AF096232), Pygidiopsis summa ({type:entrez-nucleotide,attrs:{text:AF181884,term_id:288563287,term_text:AF181884}}AF181884), and Stellantchasmus falcatus ({type:entrez-nucleotide,attrs:{text:AF181887,term_id:285804435,term_text:AF181887}}AF181887). In addition, Park [3] compared his COXI sequence of Opisthorchis viverrini Laotian isolate (AY055 382) to those of Gymnophalloides seoi ({type:entrez-nucleotide,attrs:{text:AF096234,term_id:297039736,term_text:AF096234}}AF096234) and Neodiplostomum seoulense ({type:entrez-nucleotide,attrs:{text:AF096233,term_id:285026845,term_text:AF096233}}AF096233) registered in the DNA database (Lee et al. unpublished). For the phylogenetic analyses of COXI gene of minute intestinal flukes of our own data, we have downloaded all those above mentioned COXI of Lee et al. and aligned them including our own COXI sequence of Haplorchis taichui ({type:entrez-nucleotide,attrs:{text:EF055885,term_id:119855482,term_text:EF055885}}EF055885) [4] and Paragonimus bangkokensis ({type:entrez-nucleotide,attrs:{text:AB354227,term_id:155369203,term_text:AB354227}}AB354227) [5]. Surprisingly, those sequence data were divided into 2 distinct groups without any similarities (Fig. 1). Eventually, we realized that this astonishing result is due to the reverse complementary sequences of COXI data deposited by Lee et al. (in the bottom half of the figure). We also noticed similar mixed-up deposition of the forward and reverse sequences of COXI gene of Fasciola spp., which were also included in Fig. 1 ({type:entrez-nucleotide,attrs:{text:AJ628024,term_id:88319716,term_text:AJ628024}}AJ628024, {type:entrez-nucleotide,attrs:{text:AJ628039,term_id:88319746,term_text:AJ628039}}AJ628039, {type:entrez-nucleotide,attrs:{text:FJ469984,term_id:238631966,term_text:FJ469984}}FJ469984; Zhu XQ et al. unpublished). Fig. 1 The DNA sequence alignment of the partial COXI gene of some trematodes obtained from the GenBank. Seven sequences from the top are the forward strands with JB3 primer sequence, and 2 in the middle are the forward strands without primer. Eight sequences ... For the determination of partial COX1 sequences of Platyhelminthes, the primer set of JB3 (5'-TTT TTT GGG CAT CCT GAG GTT TAT-3') and JB4.5 (5'-TAA AGA AAG AAC ATA ATG AAA ATG-3') [6] was widely used for investigating the inter- and intra-species variations of trematodes and cestodes. We noticed the mixed-up of the forward and reverse COXI sequences by Lee et al. as well as Zhu et al. because of the presence of the characteristic feature of this primer set (boxed in Fig. 1) in the sequences. The primer sequence should be deleted from the sequence data because it is not always identical with the real DNA sequence of the gene and the inclusion of the primer sequences sometimes causes the misreading in phylogenetic analyses [7]. In 3 reverse sequences, {type:entrez-nucleotide,attrs:{text:AF181884,term_id:288563287,term_text:AF181884}}AF181884, {type:entrez-nucleotide,attrs:{text:AY055380,term_id:22203992,term_text:AY055380}}AY055380, and {type:entrez-nucleotide,attrs:{text:AF096233,term_id:285026845,term_text:AF096233}}AF096233 seems to contain also the partial sequence of the cloning vector, which should be trimmed off before deposition. In general, raw data of forward and reverse sequences obtained from the sequencer should be aligned manually by cross-checking of the wave patterns because some 10-20 bases downstream from the forward primer and upstream from the reverse primer often contain erroneous base pairs [8]. Deposition of the reverse sequence means that those sequences were not aligned against forward sequence and not quite reliable. Since each sequence data in GenBank are opened for the public use, an accuracy of the sequence data is critically important for the mutual reliability of the scientists. The scientists should aware how to deposit accurate sequence data to the DNA data base. The reappraisal and correction of those sequences mentioned above is urgently necessary.

One of the Brachyura crabs found in the lower reaches of the Serayu River is Varuna litterata, also known as the "herring bow crab". This crab usually inhabits shallow tidal areas and hides under rocks, logs, or dead leaves. It is a highly adapted crab found in a wide range of salinities. V. litterata is also a fishery product consumed in Thailand, the Philippines, and Indonesia. This research was conducted in the Lower Serayu River, Central Java, which consists of three observation stations, namely: 1) the first station is located above the dam with fresh water, 2) the second station is below the dam with fresh water properties and 3) the third station is downstream (estuary) of the Serayu River with brackish water. This research was conducted for four months, from October 2023 to January 2024. This study aimed to determine the level of egg development in V. litterata. The results showed that V. litterata experienced four stages in egg development and then hatched into zoea. At the initial development stage, V. litterata eggs will be purple; in the second stage, the eggs will turn reddish; in the third development stage, the eggs will turn orange to brownish; and in the fourth stage, the eggs will develop and become black. V. litterata that will spawn will migrate to brackish waters to hatch their eggs. V. litterata produces 20,708 - 85,886 eggs with an average egg diameter of 0.440 - 0.466 mm.

ABSTRACT: This research was purposed to reveal and discover fish community in Serayu river at Banyumas Residency, Central Java, Indonesia. Research was done in two years which replicated each 3 months, since October 2009 till July 2011. Data analysis including: aquatic physical and chemical parameter and plankton species variation, fish abundance, fish species diversity and domination, age structure, condition factor, fecundity, Gonade Growth Level (GGL), Gonade Growth Index (GGI), and environmental factor and fish community structure relationship. Analysis done in this research were diverse analysis with software: Plymouth Routines in Multivariate Ecological Research (Primer), Cluster, and Multi Dimensional Scalling (MDS). During research fishes caught were as many as 3,871 fishes divided to 29 species, 20 families, and 11 ordos. Aquatic qualities factor including aquatic physical and chemical parameters and plankton species variation in Serayu river at Banyumas Residency were in good condition and suitable for fishes life. Cyprinidae family found as most varied species and most founded species in individual number at Serayu river in Banyumas Residency. There were low species diversity, but there weren’t found any dominance. Fish abundance in Serayu river increasing toward downstream, but discovered otherwise with fish diversity. Fish community in area toward upstream from Serayu Adjustable Dam differs with fish community in area toward downstream from Serayu Adjustable Dam. Serayu Adjustable Dam existence causing on decreasing fish species abundance and diversity in Serayu river at Banyumas Residency; thus, decrease fish community quality both in area before and after the dam. Fish growth pattern and reproduction profile in Banyumas Residency are not ideal. KEY WORDS: Fish community, abundance, diversity, domination, upstream and downstream, Serayu river, Serayu Adjustable Dam, and pattern and reproduction. About the Author: Dr. Susanto is a Senior Lecturer at the FKIP UMP (Faculty of Education and Teacher Training, Muhammadiyah University of Purwokerto), Jalan Raya Dukuhwaluh, Purwokerto City, Central Java, Indonesia. For academic purposes, the author can be contacted via e-mail at: susanto280266@gmail.com How to cite this article? Susanto. (2015). “Fish Community in Serayu River, Banyumas Residency, Central Java, Indonesia” in EDUCARE: International Journal for Educational Studies , Vol.8(1) August, pp…. Bandung, Indonesia: Minda Masagi Press and UMP Purwokerto, ISSN 1979-7877. Chronicle of the article: Accepted (July 15, 2015); Revised (August 3, 2015); and Published (August 25, 2015).

Abstract. Kusuma B, Prayitno SB, Sabdaningsih A, Soedibya PHT, Saputra SW. 2024. Threat of extinction of Macrobrachium esculentum in the Serayu River (Central Java, Indonesia) confirmed by DNA barcoding. Biodiversitas 25: 3531-3539. Sweet river prawn (Macrobrachium esculentum Thallwitz 1891 is one of the amphidromous shrimp. Serayu River is one of the habitats of this shrimp. Information about the M. esculentum in the Serayu River, is still minimal, so this research is essential. The Serayu Weir affects the population of Macrobrachium esculentum, which impacts the balance of the Serayu River ecosystem. This study aims to determine the condition of the M. esculentum population in the lower reaches of the Serayu River in order to maintain its sustainability and avoid the threat of extinction. This research was conducted for one year (January-December 2023). The method used was the descriptive method. We collected M. esculentum catches both upstream and downstream of the weir for one year to determine the presence of shrimp as a result of the Serayu Weir. The catch data was compared with the catch data from previous research. M. esculentum has morphological characteristics with a blackish-gray pattern with stripes along its abdomen and has an upper rostrum of 11-14 teeth and 2-4 on the lower rostrum. M. esculentum in the Serayu River has an mtDNA fragment of 686 bp and shows the same species as M. esculentum in GenBank. A total of 65 M. esculentum were collected. This shrimp was not found above the weir (Station 1) but only at station 2 (below the weir) due to the presence of weir. The results showed the importance of constructing migration routes for aquatic biota in the Serayu Weir. Further research is also needed, especially related to the reproductive system of M. esculentum.

Glyptothorax platypogon is freshwater fish, it’s potential to be an ornamental fish. Its need to be protected from extinction because population in other areas is till abundance. G. platypogon has reported that it's in Serayu River and based on pre-research observations found in Ringin River, Semarang, Central Java. The differences in the geographic location of the two rivers have the potential for the emergence of morphological variations of G. platypogon. Research on the morphology study of the G. platypogon from the Serayu River in Banjarnegara and the Ringin River has been conducted. This study describe morphological variations and provide information about the phenetic relationship between them. Sampling was carried out at Serayu River and Ringin River by direct observation and collection methods in the Field then continued at the Structure Laboratory of Biology Department State Islamic University Walisongo for observed morphometric and meristic,including 43 characters. Data was analyzed with the Kruskall Wallis test, Mann Whitney U test, PCA (Principal Component Analysis) and UPGMA (Unweighted Pair Group Arithmetic Average Method). The result showed that there were high morphological variations in the Allopatric population in Serayu River and Ringin River and there were low morphological variations in the Sympatric population in Serayu river. There were two clad taxa branching groups from Serayu and Ringin population which show morphological variation. UPGMA analysis and Euclidean distance show the population of Serayu River and Ringin River have a close phenetic relationship.

Sphaeroma terebrans, a wood-boring isopoda, is distributed worldwide in tropical and subtropical mangroves. The taxonomy of S. terebrans is usually based on morphological characteristics, with its molecular identification still poorly understood. The number of teeth on the uropodal exopod and the length of the propodus of the seventh pereopod are considered as the major morphological characteristics in S. terebrans, which can cause difficulty in regards to accurate identification. In this study, we identified S. terebrans via molecular and morphological data. Furthermore, the validity of the mitochondrial cytochrome c oxidase subunit I (COI) gene as a DNA barcode for the identification of genus Sphaeroma, including species S. terebrans, S. retrolaeve, and S. serratum, was examined. The mitochondrial COI gene sequences of all specimens were sequenced and analysed. The interspecific Kimura 2-parameter distances were higher than intraspecific distances and no intraspecific-interspecific distance overlaps were observed. In addition, genetic distance and nucleotide diversity (π) exhibited no differences within S. terebrans. Our results revealed that the mitochondrial COI gene can serve as a valid DNA barcode for the identification of S. terebrans. Furthermore, the number of teeth on the uropodal exopod and the length of the propodus of the seventh pereopod were found to be unreliable taxonomic characteristics for S. terebrans.

BackgroundTaxonomic uncertainties in the morphological species identification and taxonomic revisions in individual groups are known for all echinoderm classes. These uncertainties in morphological species identification and discrimination have spawned the application of molecular genetic identification techniques. However, as the fundamental step to allow and ensure future molecular species identification, valid and comprehensive reference library entries comprising morphological and molecular species information together with various metadata are essentially needed. In our study we compare morphological and molecular genetic species identification techniques for representatives of North Sea echinoderm classes, i.e. the Asteroidea, Ophiuroidea, Echinoidea and Holothuroidea.MethodsIndividuals were sampled during different surveys in different regions of the North Sea, identified to species level based on morphological diagnostic features, and were genetically analysed using a fragment of the mitochondrial cytochrome c oxidase subunit I (COI).Results and DiscussionThe morphological determination revealed 32 species including one taxon determined only to genus level. In contrast to this, the COI analysis supported 34 monophyletic clades with pronounced differences between the intra- and the inter-specific genetic variability (a barcoding gap of 4.93 %) with highest intra-specific variabilities found in the ophiuroid species Amphiura filiformis, A. chiajei and Ophiura sarsii. In 94 % of the investigated species, morphological identification and COI sequence clusters were congruent whereas for two asteroid species we found an underestimated diversity. For Astropecten irregularis, one of the most common starfish species of the North Sea, we found two distinct and possibly depth-related clades, probably sibling species, differing by 11.1–11.9 % sequence divergences (p-distances). For two starfish individuals, morphologically identified as Henricia sanguinolenta, the COI analysis revealed two monophyletic clades, of which one was classified as H. cf. oculata by comparison to published sequences.ConclusionsThis newly established sequence reference library for the North Sea Echinodermata allows and ensures future molecular species identification for various life-cycle stages including juveniles and meroplanktonic larvae and provides sequences for phylogeographic studies and the detection of sibling as well as cryptic species.

Co-overexpression of the epidermal growth factor (EGF) receptor (EGFR) and c-Src frequently occurs in human tumors and is linked to enhanced tumor growth. In experimental systems this synergistic growth requires EGF-dependent association of c-Src with the EGFR and phosphorylation of Tyr-845 of the receptor by c-Src. A search for signaling mediators of Tyr(P)-845 revealed that mitochondrial cytochrome c oxidase subunit II (CoxII) binds EGFR in a Tyr(P)-845- and EGF-dependent manner. In cells this association involves translocation of EGFR to the mitochondria, but regulation of this process is ill-defined. The current study demonstrates that c-Src translocates to the mitochondria with similar kinetics as EGFR and that the catalytic activity of EGFR and c-Src as well as endocytosis and a mitochondrial localization signal are required for these events. CoxII can be phosphorylated by EGFR and c-Src, and EGF stimulation reduces Cox activity and cellular ATP, an event that is dependent in large part on EGFR localized to the mitochondria. These findings suggest EGFR plays a novel role in modulating mitochondrial function via its association with, and modification of CoxII.

Characteristic of cytochrome C oxidase subunit I (COI) gene in giant clam from Wakatobi National Park Waters was studied to detect the specific nucleotide and show the relationship among species of giant clams. Sample collection was conducted around Wangi-wangi, Kaledupa, and Tomia waters. Genomic DNA was extracted from clam mantle tissue of two generas, and amplified by PCR method. We obtained nucleotide size of 522 bp for genus Tridacna (T. crocea, T. maxima) and 708 bp for genus Hippopus (H. hippopus). There were 52 specific nucleotide for the clams (included T. gigas and T. squamosa from GenBank data). T. crocea, T. gigas, T. squamosa, T. maxima, and H. hippopus had 2, 3, 9, 2, and 36 sites, respectively. Reconstruction phylogenetic tree using Neighbour Joining of 476 bp COI gene sequences showed that all species of giant clams were monophyletic.

Belodontichthys dinema with the local name of sengarat fish, is one of freshwater fish in Riau Province of Indonesia. B. dinema distributed in some rivers of Riau Province were Indragiri and Tapung rivers. This study reports analysis of DNA barcode of B. dinema using cytochrome c oxidase subunit I (COI) gene. The muscle of B. dinema fish using for DNA extraction. In this study had been obtained 652 bp of COI gene from Polymerase Chain Reaction (PCR) product. The Basic Local Alignment Search Tool (BLAST) analysis based on COI gene sequences shows that B. dinema from the Indragiri and Tapung rivers has 100% similarity with B. dinema from the existing database in GenBank. There are 14 marker nucleotides can characterize the species B. dinema molecularly. The value of genetic distance between B. dinema from Indragiri and Tapung rivers with B. dinema from GenBank data is 0.00. The phylogenetic tree showed that B. dinema from Indragiri and Tapung rivers, formed one group with B. dinema from Genbank data with bootstrap value is 100%. These results confirmed that sengarat fish identified was B. dinema based on the COI gene as DNA barcode.

The Satyrinae is a subfamily of Nymphalid butterfly, which is morphologically and ecologically the most diverse group, occurring in all habitats. In the present study, Cytochrome c oxidase subunit I (COI) gene of seven species of Satyrinae was sequenced, aligned, and used to construct phylogenetic trees. The molecular identification of these Satyrinae species was confirmed by comparing the related sequences in the National Center for Biotechnology Information (NCBI) GenBank. The base compositions of the COI sequences were 39.07% T, 16.44% C, 29.83% A, and 14.64% G, revealing a strong AT bias (68.9%). The sequence distance among Satyrinae species ranged from 0.09% to 0.18%. Phylogenetic trees were constructed by the neighbor-joining (NJ) and maximum likelihood (ML) methods, using Orthetrum sabina as an outgroup. Both trees had almost identical topologies. The sampled species in Satyrinae exhibited the following relationships: Melanitis leda + [(Mycalesis mineus+(Mycalesis gotama+Mycalesis anaxias)) + (Ypthima baldus + (Lethe chandica+Elymnias hypermnestra))], suggesting that M. leda might be distantly related with the rest of the Satyrinae species. This clustering result is almost identical to current traditional classification. This study confirms that the COI based DNA barcoding is an efficient method for the identification of butterflies including Satyrinae species and, as such, may further contribute effectively to biodiversity and evolutionary research.
 Jahangirnagar University J. Biol. Sci. 8(1): 67-74, 2019 (June)

Background and Aim:Q fever Coxiella burnetii is a worldwide zoonotic disease, and C. burnetii was detected in mammals and ticks. Ticks play an important role in the spread of C. burnetii in the environment. Therefore, the aims of this study were to detect Q fever C. burnetii in camels and ixodid ticks by molecular tools and identification of Hyalomma dromedarii and Hyalomma excavatum using molecular and immunological assays.Materials and Methods:A total of 113 blood samples from camels and 190 adult ticks were investigated for the infection with C. burnetii by polymerase chain reaction (PCR) and sequencing the targeting IS30A spacer. The two tick species H. dromedarii and H. excavatum were characterized molecularly by PCR and sequencing of 16S ribosomal RNA (16S rRNA) and cytochrome oxidase subunit-1 (CO1) genes and immunologically by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and western blot.Results:A total of 52 camels (46%) were positive for Q fever infection. Only 10 adult ticks of H. dromedarii were infected with C. burnetii. The IS30A sequence was around 200 bp in length for C. burnetii in H. dromedarii ticks with a similarity of 99% when compared with reference data in GenBank records. The length of 16S rDNA and CO1 was 440 and 850 bp, respectively, for both H. dromedarii and H. excavatum. The phylogenetic status of H. dromedarii was distant from that of H. excavatum. SDS-PAGE revealed seven different bands in the adult antigens of either H. dromedarii or H. excavatum with molecular weights ranged from 132.9 to 17.7 KDa. In western blot analyses, the sera obtained from either infested camel by H. dromedarii or infested cattle by H. excavatum recognized four immunogenic bands (100.7, 49.7, 43.9, and 39.6 kDa) in H. dromedarii antigen. However, the infested camel sera identified two immunogenic bands (117 and 61.4 kDa) in H. excavatum antigen. Furthermore, the sera collected from cattle infested by H. excavatum recognized three immunogenic bands (61.4, 47.3, and 35 kDa) in H. excavatum antigen.Conclusion:Molecular analyses indicated that both camels and ticks could be sources for infection of animals and humans with Q fever. Furthermore, the molecular analyses are more accurate tools for discriminating H. dromedarii and H. excavatum than immunological tools.

Species in the brackish crab genus Ptychognathus are common in the seashore and estuary habitats with freshwater input. Due to their similar morphologies and dull colorations, it is always difficult to distinguish the species in this genus. In this study, the DNA barcode gene COI (cytochrome c oxidase subunit I) was used to help identify Ptychognathus from Taiwan. The results showed that the 10 species can be identified successfully based on COI, with intraspecific distances below 1.54% and interspecific distances of 12.2%-19.57%. In addition, two new species of Ptychognathus are described from Taiwan. Ptychognathus makii sp. nov. from southern Taiwan is similar to P. altimanus (Rathbun, 1914), and P. stimpsoni sp. nov. from southern Taiwan and the southern Philippines resembles P. aff. barbatus (A. Milne-Edwards, 1873) and P. pusillus Heller, 1865. Both species can be distinguished from other congeners by a suite of characters, including the carapace, orbital region, frontal region, telson of male pleon, male first gonopod, and setae on ambulatory legs.

Among the Ixodid ticks, Hyalomma anatolicum is a well-known vector that transmits various pathogens to terrestrial and semi-terrestrial vertebrates including humans, and its population differ in ecology and vector competence. Expansion of this tick to new areas changes the genetic structure, and lead to affect the vector-pathogen interaction and disease outcomes. The present study was designed to infer the haplotype diversity, demographic dynamics, gene flow and genetic differentiation, and phylogeny of H. anatolicum from different countries based on the cytochrome oxidase I (COI) and 16S rDNA sequences. A total of 320 ticks were collected from cattle, buffaloes, and sheep in five districts of Khyber Pakhtunkhwa, Pakistan, morphologically identified as H. anatolicum, and subjected to genetic analysis. A total 85 and 138 sequences for COI and 16S rDNA, including 11 and 2 sequences generated in this study, respectively, were analyzed to assess haplotype network, population structure and divergence, demographic changes, and phylogenetic analysis. Analysis based on COI sequences yielded 29 haplotypes in which haplotype 1 and 15 were the predominant consisting of 35 and 20 sequences, respectively, from Pakistan, India, China, Bangladesh, Iraq, Saudi Arabia, Kazakhstan and Egypt. The 16S rRNA yielded 30 haplotypes in which haplotype 1 was predominant consisting of total 86 sequences from Pakistan, India, China, United Arab Emirates, Tajikistan, Kazakhstan, Turkey, Egypt, and Iraq. Complete haplotype network based on COI and 16S rRNA confirmed stellate structure, together with high haplotype diversity (COI 0.77899, 16S rRNA 0.60774) and low nucleotide diversity (COI 0.00445, 16S rRNA 0.00431), which support recent population expansion. Similarly, neutrality indices for the whole dataset, Tajima’s D (COI − 2.36363**, 16S rRNA − 2.54127***), Fu and Li’s D (COI − 5.72992, 16S rRNA − 6.31313*), and Fu and Li’s F (COI − 5.04435*, 16S rRNA − 5.56085*) were negative, indicating deviation from neutrality and recent population dispersal. In the phylogenetic tree based on the COI and 16S rDNA sequences, with exception of one sequence for a single haplotypes, which appeared independently, there is a single main clade that includes the largest number of sequences for all other haplotype. Based on COI and 16S rDNA sequences, the present study provided first detail information about the population genetics and haplotype networks of H. anatolicum.