COX-1 Research Articles

Morphometric and Molecular Characterization of Fasciola spp. in Livestock From Northwestern Provinces of Iran.

Liver flukes from the genus Fasciola are the causative agents for human and livestock fascioliasis. Accurate identification of Fasciola spp. is essential to understanding the epidemiology of fascioliasis. This study aimed to determine the morphometric and molecular characterization of Fasciola spp. in livestock from Northwestern provinces of Iran. Five hundred adult Fasciola flukes were obtained from different definitive hosts (cattle, sheep, goats, and buffaloes) in four local abattoirs in the northwestern provinces of Iran (West-Azerbaijan, East-Azerbaijan, Ardabil, and Zanjan) from September 2021 to August 2022. All samples were examined by morphometric criteria; then, 49 samples were identified using PCR-RFLP based on ITS1 region and 23 sequence of isolates analyzed by cox1 marker. PCR-RFLP methods compared morphometric results, and cox1 gene sequences were used to confirm PCR-RFLP results and phylogenetic analysis. The differences between the body length, body width, cephalic cone length, cephalic cone width, body area, and distance between the ventral sucker and posterior end of the body in two species were significant (p < 0.05). Based on the morphometric criteria, 139 samples (27.8%) were identified as Fasciola gigantica and 361 (72.2%) as Fasciola hepatica. Similarly, PCR-RFLP analysis of ITS1 region confirmed morphometric results. No hybrid forms of Fasciola were detected. Partial sequences of cox1 showed 13 variable sites with eight haplotypes in F. hepatica and 12 variable sites with five haplotypes in F. gigantica. Based on the results of this study, the PCR-RFLP method can be used to confirm the morphological method of Fasciola species, but it is insufficient to study their genetic diversity. Also, sequences of cox1 results of the present study showed that F. hepatica and F. gigantica species in the Northern provinces of Iran have different genetic structures and haplotypes.

Molecular characterization of Fasciola hepatica obtained from cattle and horse in Central Chile

Liver fluke infection, caused by the trematode Fasciola hepatica, is a parasitic zoonotic disease affecting various mammals, including humans, and has significant implications for public, animal, and ecosystem health. This study provides the first genetic characterization of F. hepatica in Chile, focusing on the complete mitochondrial gene cox1. Samples were collected from two different host species: cattle and horses. Our findings revealed that 70 % of detected haplotypes were found in either cattle or horses, which coincides with their geographical origin. Interestingly, the use of full-length sequences resulted in the identification of 80 % unique sequences, whereas this reduced to 45 % when analyzing the traditionally used short sequences. This underestimation of genetic diversity suggests that broader sequencing efforts might be essential for a more accurate understanding of F. hepatica genetic landscape. This research underscores the importance of understanding the genetic variability in parasites to improve strategies for disease control and treatment.

Anisakis spp. as indicators of latitudinal movements of short‐finned pilot whales, Globicephala macrorhynchus, in the Northeast Atlantic

AbstractIn the Northeast Atlantic, it is unclear whether short‐finned pilot whales, Globicephala macrorhynchus (SFPWs), are transient or resident above 40°N. We used Anisakis spp. to identify the latitudes recently visited by a SFPW pod stranded in NW Spain (43°N) in 2020. Analysis of cox2 gene in 30 nematodes from 6 SFPWs revealed the presence of A. simplex sensu stricto (s.s.) (93.3%) and A. pegreffii (6.7%). Morphological analysis of 972 nematodes corroborated species molecular identification and relative proportions; in males (n = 66), the estimated proportions of A. simplex s.s. and A. pegreffii were 86.4% and 13.6%, respectively. These percentages resembled those reported in fish/cetaceans above 40°N, and L4 larvae or adults of A. typica (a tropical‐temperate species found in SFPWs up to 38°N–39°N) were not detected. Population structure of A. simplex s.s. + A. pegreffii in SFPWs suggested a continuous recruitment of nematodes starting at least 3 weeks before stranding. We interpret that either the SFPW pod was within the range of the species (perhaps as a recent northern shift) or represented a vagrant group that visited waters off Northwest Spain for a protracted period. Future analysis on nematode assemblages could shed light on movements and climate‐driven shifts in cetacean distribution.

Comparative selective pressure analysis on mitochondrial protein-coding genes in flying squirrels (Pteromyini) and tree squirrels (Sciurini)

Different animal groups with varying locomotion modes may have unique energy requirements. Mitochondria produce adenosine triphosphate (ATP) and reactive oxygen species via oxidative phosphorylation to support organisms energy requirements. The tribes Pteromyini (flying squirrels) and Sciurini (tree squirrels), two closely related taxa within the family Sciuridae, exhibit distinct locomotion modes, energy requirements, and likely face different selective pressures on mitochondrial protein-coding genes (PCGs). We analysed 13 mitochondrial genome sequences from species belonging to the tribe Pteromyini and 117 from species belonging to the tribe Sciurini. Phylogenetic analysis revealed Pteromyini and Sciurini formed a sister relationship within the family Sciuridae. Among the 13 PCGs, ATP8 exhibited the highest dN/dS values, while COX1 showed the lowest. The background selection ratio (ω2) values for six genes (ND1, ND2, ND4, ATP6, ND5, and COX3) in Pteromyini were lower than the foreground selection ratio (ω0) values observed in Sciurini. A RELAX analysis revealed that CYTB, ND4, ATP6, and COX3 genes experienced intensified in selection strength. BUSTED analysis identified stronger signatures of diversifying selection in CYTB and ATP6, highlighting amino acid changes. MEME identified episodic diversifying selection at specific sites among eight PCGs. These findings revealed distinct selective pressures on PCGs in flying and tree squirrels.

Molecular characterization and genetic variability of Toxocara vitulorum from naturally infected buffalo calves for the first time in Bangladesh.

Toxocara vitulorum is one of the deadliest parasite of buffalo calves in Bangladesh. This study was conducted to explore genetic variability within and among the T. vitulorum populations in buffalo calves of Bangladesh. Genomic DNA was extracted, ITS2, COX1 and NAD1 gene were amplified and sequenced. Distinct 29 ITS2, 21 unique NAD1 and 24 COX1 genotypes were detected among the T. vitulorum of different geographic regions. These three gene genotypes similarities ranged from 97 to 99%, when these were compared to best hit scoring T. vitulorum sequences retrieved from GenBank. A total of 12 and 6 unique haplotypes were detected for COX1 and NAD1 gene sequences. The average nucleotide and haplotype diversity for COX1 and NAD1 were 0.0931 & 0.89493 and 0.00658 & 0.77895 respectively and the recorded values were more dispersed than previously published values. The pairwise Nst values ranged from −0.050 to 0.602 and Fst from −0.050 to 0.600 between all the T. vitulorum genotypes indicated huge genetic differentiation which were reportedly higher than other published reports Fst values. This is the first report of T. vitulorum on the basis of COX1 gene in Bangladesh. The study findings will be helpful for further extensive epidemiological studies regarding anthelmintic resistance, control and prevention of T. vitulorum infection in buffalo calves.

The terrestrial flatworm Microplana scharffi (Geoplanidae, Microplaninae): mitochondrial genome, phylogenetic proximity to the Bipaliinae and genes related to regeneration

A genome skimming approach of sequencing was undertaken on a subfamily of terrestrial flatworms that had been neglected in genomic studies until now, namely the Microplaninae as represented here by Microplana scharffi. A single run of short-read sequencing enabled retrieval of the complete mitogenome, the two paralogous versions of the 18S gene, the elongation factor gene EF1α, plus two genes involved in the regeneration process, namely those coding for ß-CATENIN-1 and adenomatous polyposis coli (APC). The 15,297 bp mitogenome lacks a functional tRNA-Ala and has a mandatory alternative TTG start codon in its cox1 gene. The multiprotein phylogeny, inferred from mitogenome proteins, positions M. scharffi as sister-group to the Bipaliinae with maximum support, although the organisation of the mitogenomes shows features previously never observed among Bipaliinae, such as the conserved 32 bp overlap between ND4 and ND4L. Similarly to what has been observed in recent publications on other species of Geoplanidae, the two types of 18S genes display strongly different coverages and are only 90.57% identical. Additionally, alien DNA was identified in the pool of contigs in the form of the complete mitochondrial genome of Lumbricus rubellus, confirming previous observations on the feeding habits of M. scharffi.

Human Schistosomiasis due to Schistosoma bovis in Nigeria.

Schistosomiasis is a global public health challenge, especially in sub-Saharan Africa, where Nigeria has the highest burden of disease. Schistosoma hybrids have been discovered in various countries, including Nigeria, where livestock and human beings share common water resources. This study, carried out in three communities, two of which are endemic for urinary schistosomiasis, aimed to identify the urinary schistosomiasis causative agent by using a species-specific molecular technique and their evolutionary relationship. Polymerase chain reaction using primers specific for a partial DNA sequence of the mitochondrial cox1 gene of Schistosoma haematobium was carried out on pooled urine sediments preserved in 70% ethanol. DNA from the high-intensity pooled samples from Onye-Uku camp amplified, and a BLAST search identified the pooled samples as Schistosoma bovis, whereas S. haematobium DNA did not amplify. The phylogenetic relationship of the sequence showed that it clustered with an S. bovis hybrid obtained from a human host in Côte d'Ivoire and had close ancestry with isolates from cattle in Cameroon. This finding revealed the prevalence of S. bovis among some inhabitants in a zone in Nigeria where schistosomiasis is endemic.

First Molecular Identification and Prevalence of Sarcocystis spp. in Sheep Intended for Human Consumption in Shanxi Province, China.

Sarcocystis species are intracellular coccidian protozoans that can infect a range of animals and humans and cause public health problems and economically significant losses. Sarcocystosis in sheep (Ovis aries) can cause abortion, neurological symptoms, and even death and results in significant economic losses to the livestock industry. To date, however, it is yet unknown whether sheep in Shanxi Province, north China, are infected with Sarcocystis spp. The purpose of this study was to investigate the prevalence of Sarcocystis spp. in sheep in Shanxi Province. Thus, 582 muscle samples of sheep were purchased from farmers' markets from ten representative counties in Shanxi Province, north China, and examined for the presence and prevalence of Sarcocystis spp. by PCR amplification of the mitochondrial cytochrome c oxidase subunit I (cox1) gene. Of the examined 582 mutton samples, 197 samples (33.85%) were Sarcocystis-positive and were sequenced. Of the obtained 197 cox1 sequences, 196 sequences showed nucleotide similarity of 98.56-99.81% with those of S. tenella, and the remaining one cox1 sequence showed nucleotide similarity of 99.71% with that of S. arieticanis. Two representative cox1 sequences of S. tenella (accession nos. PQ189447 and PQ189448) have 99.52% and 99.61% identity with S. tenalla (KC209725) and S. tenalla (MK419984), respectively. The sequence of S. arieticanis (accession no. PQ165949) obtained in this study has 99.71% identity with S. arieticanis (MK419975). This present study documents the occurrence and prevalence of Sarcocystis spp. in sheep in Shanxi Province for the first time, which enriches the data on the distribution of Sarcocystis spp. in sheep in China and has implications for the control of sheep sarcocystosis.

Prevalence and genetic diversity of the lung nematode Eucoleus aerophilus in red foxes (Vulpes vulpes) in Central Europe (Poland) assessed by PCR and flotation

Eucoleus aerophilus is the causative agent of respiratory capillariosis in wild and domestic carnivores and has sporadically affected humans. To date, this parasite has been detected in 38 countries, confirming its worldwide distribution. The aim of this study was to evaluate the prevalence of E. aerophilus in red fox populations from Central Europe and to describe the sequence variations in the partial cox1 gene of this parasite recovered from stool samples. An investigation was carried out using 342 samples of red foxes faeces investigated via seminested PCR and coproscopy. PCR results confirmed the presence of E. aerophilus DNA in 230 samples and E. boehmi DNA in 14 samples. Molecular analysis of the retrieved sequences revealed 22 haplotypes of E. aerophilus, EaPL1–EaPL22, and 4 haplotypes of E. boehmi, EbPL1–EbPL4. Coproscopic examination revealed that eggs of the Capillariidae family were most prevalent in all regions, with a mean prevalence of 73%. Furthermore, the following eggs were detected: Taeniidae (21.3%), Toxocara spp. (26.3%), Toxascaris leonina (5.3%), Trichuris vulpis (1.5%), trematodes (23.4%), hookworms (2.3%) and Mesocestoides spp. (1.8%). This study evaluated the prevalence of E. aerophilus in red fox populations in selected regions of Poland. To the best of our knowledge, this is the first molecular study on E. aerophilus in Poland.

Comparative Mitogenomics Analysis Revealed Evolutionary Divergence among Purpureocillium Species and Gene Arrangement and Intron Dynamics of Ophiocordycipitaceae.

The species of Purpureocillium are cosmopolitan and multitrophic fungi that can infect a wide range of invertebrate hosts. This study reports the mitogenome of P. atypicola, a specialized spider pathogenic fungus. The 112,465 bp mitogenome encoded genes typically found in fungal mitogenomes, and a total of 52 introns inserted into seven genes. A comparison with three other Purpureocillium species revealed significant differences in length and intron number, primarily due to intron variation; however, there was no dynamic variation in the introns of the cox1 gene within the same species of the Purpureocillium genus. Different mitochondrial protein-coding genes showed variable degrees of genetic differentiation among these species, but they were all under purifying selection. Additionally, frequent intron loss or gain events were detected to have occurred during the evolution of the Ophiocordycipitaceae mitogenomes, yet the gene arrangement remains conserved. A phylogenetic analysis of the combined mitochondrial gene set gave identical and well-supported tree topologies. The estimated age of the crown of Ophiocordycipitaceae and Purpureocillium were around the Early Cretaceous period (127 Mya) and Late Cretaceous period (83 Mya), respectively. The results of this study advance our understanding of the genomics, evolution, and taxonomy of this important fungal group.

Molecular detection of Coxiella burnetii in blood and hard tick-infested Egyptian camels and the possibility of coinfections

Coxiella burnetii, a bacterium that causes Q fever. It can infect mammals and has a global geographical distribution, but data on its occurrence in Egyptian dromedaries and the associated ticks are limited. Therefore, this study aims to detect C. burnetii in the blood of infested camels and associated ticks collected from Egypt by using molecular techniques and to examine the possibility of coinfections with C. burnetii. A total of 133 blood samples and 1260 hard ticks infesting these camels were collected from Egyptian slaughterhouses. Nested PCR and sequencing were used based on the IS1111 gene for molecular detection of C. burnetii. The identification of tick species at the molecular level was performed using the COX1 gene. C. burnetii was detected in Hyalomma (H) dromedarii, H. anatolicum, H. marginatum, Amblyomma (Am) lipidium, and Am. cohaerens with an overall prevalence rate of 1.3% (16/1260), while in the camel blood samples, it was 15.8% (21/133). Out of C. burnetii-positive ticks, there were double infections by Borrelia species and C. burnetii in H. dromedarii and Am. lipidium and triple infections at one Am. cohaerens tick (C. burnetii, Borrelia spp., and Babesia microti). In addition, two positive camel blood samples were found to carry C. burnetii with Borrelia spp. Our research findings indicate the presence of Coxiella burnetii among camels and their associated ticks in Egypt and emphasize the potential of having coinfection. To prevent the transmission of this infection to other animal species or humans, appropriate control measures should be implemented.

Aqueous Extracts of Rhus trilobata Inhibit the Lipopolysaccharide-Induced Inflammatory Response In Vitro and In Vivo.

Chronic noncommunicable diseases (NCDs) are responsible for approximately 74% of deaths globally. Medicinal plants have traditionally been used to treat NCDs, including diabetes, cancer, and rheumatic diseases, and are a source of anti-inflammatory compounds. This study aimed to evaluate the anti-inflammatory effects of Rhus trilobata (Rt) extracts and fractions in lipopolysaccharide (LPS)-induced inflammation models in vitro and in vivo. The aqueous extract (RtAE) and five fractions (F2 to F6) were obtained via C18 solid-phase separation and tested in murine LPS-induced J774.1 macrophages. Key inflammatory markers, such as IL-1β, IL-6, TNF-α, and COX-2 gene expression were measured using RT-qPCR, and PGE2 production was assessed via HPLC-DAD. The in vivo effects were tested in an LPS-induced paw edema model in Wistar rats. Results showed that RtAE at 15 μg/mL significantly decreased IL-1β and IL-6 gene expression in vitro. Fraction F6 further reduced IL-1β, TNF-α, and IL-6 gene expression, COX-2 expression, and PGE2 production. In vivo, F6 significantly reduced LPS-induced paw edema, inflammatory infiltration, and IL-1β and COX-2 protein expression. Chemical characterization of F6 by UPLC/MS-QTOF revealed at least eight compounds with anti-inflammatory activity. These findings support the anti-inflammatory potential of RtAE and F6, reinforcing the medicinal use of Rt.

Genetic Insights into the Giant Keyhole Limpet (Megathura crenulata), an Eastern Pacific Coastal Endemic: Complete Mitogenome, Phylogenetics, Phylogeography, and Historical Demography

Background: The giant keyhole limpet Megathura crenulata is a gastropod mollusk (Fissurella superfamily) that is endemic to the eastern Pacific coast from southern California, USA, to Baja California Sur, Mexico. M. crenulata is socioeconomically important as it produces a potent immune-stimulating protein, called Keyhole Limpet Hemocyanin, which is extracted in vivo and utilized for vaccine development. However, ecological studies are scarce and genetic knowledge of the species needs to be improved. Our objectives were to assemble and annotate the mitogenome of M. crenulata, and to assess its phylogenetic relationships with other marine gastropods and to evaluate its population genetic diversity and structure. Methods: Samples were collected for mitogenome assembly (n = 3) spanning its geographic range, Puerto Canoas (PCA) and Punta Eugenia (PEU), Mexico, and California (CAL), USA. Total DNA was extracted from gills sequenced using Illumina paired-end 150-bp-read sequencing. Reads were cleaned, trimmed, assembled de novo, and annotated. In addition, 125 samples from eight locations were analyzed for genetic diversity and structure analysis at the 16s rRNA and COX1 genes. Results: The M. crenulata mitogenomes had lengths of 16,788 bp (PCA) and 16,787 bp (PEU) and were composed of 13 protein-coding regions, 22 tRNAs, two rRNAs, and the D-Loop region. In terms of phylogeographic diversity and structure, we found a panmictic population that has experienced recent demographic expansion with low nucleotide diversity (0.002), high haplotypic diversity (0.915), and low φST (0.047). Conclusions: Genetic insights into the giant keyhole limpet provides tools for its management and conservation by delimiting fishing regions with low genetic diversity and/or genetically discrete units.

Exploring transmission dynamics of the Sarcoptes scabiei mite in humans by combining molecular typing and epidemiological variables, the Netherlands 2016–2023

BackgroundScabies, an infestation of the mite Sarcoptes scabiei, has seen an increase in clinical diagnoses in the Netherlands since 2011. This study aimed to analyse PCR-positive S. scabiei skin samples through partial genome sequencing and to link findings to patient epidemiological characteristics.MethodsSkin samples were collected from individuals in the Netherlands between January 2016 and January 2023. On the PCR-positive S. scabiei skin samples, partial mitochondrial cytochrome c oxidase subunit 1 gene (cox1) sequencing was performed to assess genetic variability. Epidemiological information was collected through interviews. We examined associations between cox1 subtypes, epidemiological factors and treatment outcomes.ResultsSequencing results were obtained from 128 patients, with epidemiological information available for 55 (43%) of these patients. Fifteen distinct cox1 subtypes were identified. Subtype 01 was most prevalent (45%) and present across all age groups and social settings. The remaining subtypes were less common and not consistently found in all contexts. Five clusters were identified, each with identical cox1 subtypes. Comparative analysis with GenBank sequences revealed genetic similarities with strains from Australia, the USA and China, suggesting the global distribution and transmission of specific subtypes. A substantial proportion (73%) of patients with scabies required multiple treatments to eradicate the infestation, with no subtype-related differences.ConclusionsThis is the first study linking S. scabiei sequencing results to patient epidemiological data. Several subtypes clustered in specific geographic regions and social contexts, underscoring localised transmission patterns. Further research with larger sample sizes is needed to enhance our understanding of the transmission of this mite. This study provides valuable insights that will strengthen scabies control efforts.Graphical

Investigating the effects of combined treatment of mesalazine with Lactobacillus casei in the experimental model of ulcerative colitis.

Ulcerative colitis (UC), a common gastrointestinal disorder in affluent nations, involves chronic intestinal mucosal inflammation. This research investigated the effects of combined probiotic treatment of Lactobacillus casei (L. casei) and mesalazine on disease activity index and inflammatory factors in the UC model. 20 male BALB/c mice were utilized and divided into four groups. To induce UC, all groups received 100 μL of 4% acetic acid (AA) intra-rectally. The first group received phosphate-buffered saline (PBS) (as a control group), the second group was treated with L. casei, the third group was treated with mesalazine and, the fourth group was treated with L. casei and mesalazine. Treatment with L. Casei and mesalazine commenced after the manifestation of symptoms resulting from UC induction. Finally, the mice were euthanized and the disease activity index, myeloperoxidase activity, nitric oxide rate, cytokines level (IL-1β, IL-6, TNF-α) and, gene expression (iNOS, COX-2, and cytokines) were evaluated. The combined treatment of L. casei and mesalazine led to a significant decrease in the levels of NO, MPO and inflammatory cytokines. In addition, the expression of cytokines, iNOS and COX-2 genes decreased in mice treated with the combination. This study shows that combined treatment of L. casei and mesalazine improves of experimental UC, which can be attributed to the anti-inflammatory properties of L. casei and mesalazine. In conclusion, this combination therapy can be considered a suitable option for the management of UC.

Development of a New Isoxsuprine Hydrochloride-Based Hydroxylated Compound with Potent Antioxidant and Anti-Inflammatory Activities.

The scientific community actively pursuits novel compounds with biological activities. In this context, our study utilized the predicted data mining approach (PDMA), which can efficiently screen out biotransformable precursor candidates to produce new bioactive compounds. The PDMA was applied to Bacillus megaterium tyrosinase (BmTYR) to form new bioactive hydroxyl compounds from isoxsuprine hydrochloride (isoxsuprine). The results show that isoxsuprine could be biotransformed by BmTYR to form a new compound, 3''-hydroxyisoxsuprine. 3''-Hydroxyisoxsuprine exhibited 40- fold and 10-fold higher potent antioxidant and anti-inflammation activities than the precursor, isoxsuprine. The 3''-hydroxyisoxsuprine effectively mitigates the hyperimmune response in RAW 264.7 macrophages by inhibiting the upregulation of pro-inflammatory cytokine (IL-1β and IL-6) and inflammatory enzyme COX-2 gene expression triggered by LPS stimulation. This study illustrates that PDMA is an effective strategy for screening known natural and chemical compounds and for generating new bioactive compounds through biotransformation. Our newly produced compound has potential future applications in pharmacology and biotechnology.

Cloning, characterization of β-glucosidase from Furfurilactobacillus rossiae in bioconversion and its efficacy.

Minor ginsenosides produced by β-glucosidase are interesting biologically and pharmacologically. In this study, new ginsenoside-hydrolyzing glycosidase from Furfurilactobacillus rossiae DCYL3 was cloned and expressed in Escherichia coli strain BL21. The enzyme converted Rb1 and Gyp XVII into Rd and compound K following the pathways: Rb1→Rd and Gyp XVII→F2→CK, respectively at optimal condition: 40°C, 15min, and pH 6.0. Furthermore, we examined the cytotoxicity, NO production, ROS generation, and gene expression of Gynostemma extract (GE) and bioconverted Gynostemma extract (BGE) in vitro against A549 cell lines for human lung cancer and macrophage RAW 264.7 cells for antiinflammation, respectively. As a result, BGE demonstrated significantly greater toxicity than GE against lung cancer at a dose of 500µg/mL but in normal cells showed lower toxicity. Then, we indicated an enhanced generation of ROS, which may be boosting cancer cell toxicity. By blocking the intrinsic way, BGE increased p53, Bax, Caspase 3, 9, and while Bcl2 is decreased. At 500µg/mL, the BGE sample was less toxic in normal cells and decreased the LPS-treated NO and ROS level to reduce inflammation. In addition, BGE inhibited the expression of pro-inflammatory genes COX-2, iNOS, IL-6, and IL-8 in RAW 264.7 cells than the sample of GE. In conclusion, FrBGL3 has considerable downstream applications for high-yield, low-cost, effective manufacture of minor ginsenosides. Moreover, the study's findings imply that BGE would be potential materials for anti-cancer and anti-inflammatory agent after consideration of future studies.

Microcotyle Tazeroutii n. sp. (Monogenea: Microcotylidae) from the Gills of the Boarfish Capros aper Linnaeus, 1758 (Teleostei: Caproidae) off the Algerian Coast, Western Mediterranean.

In this study we describe a new species Microcotyle tazeroutii n. sp. (Monogenea: Microcotylidae) found on the gills and operculum of the boarfish Capros aper (Caproidae) off the Algerian coast of the Western Mediterranean. Monogeneans were observed alive or recently dead on the operculum and gills using a dissecting microscope, measured and drawn for morphological study. Furthermore, a molecular analysis was conducted using a partial fragment of the mitochondrial cytochrome c oxidase subunit 1 (cox1) of two specimens of monogeneans and a tissue sample of the fish's gills in which the parasites were found to confirm the identity of fish. The new species Microcotyle tazeroutii n. sp., exhibits a combination of morphological features that differentiate it from all other known species within the genus, such as the shape and the size of body, the haptor length, the number and the size of clamps and testes, the number of spines of the genital atrium and the size of eggs. Additionally, a molecular analysis of the mitochondrial cytochrome c oxidase subunit 1 (cox1 gene) revealed significant interspecific differences between Microcotyle tazeroutii n. sp. and previously published sequences of other Microcotyle species. The morphological and molecular analyses revealed that Microcotyle tazeroutii n. sp. has unique characteristics that distinguish it from all previously identified species and confirmed the presence of Microcotyle within the Caproidae family for the first time.

First Report of Haemaphysalis longicornis (Neumann) in Oklahoma, USA.

Haemaphysalis longicornis (Neumann), the Asian longhorned tick, is a species native to East Asia, but invasive to Australia, New Zealand, and most recently, the United States. It has spread rapidly across the eastern United States after being established in New Jersey in 2017. Aiding this rapid expansion is the ability of this tick to reproduce parthenogenically and feed on diverse host species. In cattle, this tick can cause heavy burdens and act as a vector for the pathogenic hemoprotozoan parasite Theileria orientalis, genotype Ikeda, creating economic losses that impact the cattle industry. Here, we report Asian longhorned ticks, collected from cattle, a dog, and pastures and morphologically identified at the Oklahoma Animal Disease Diagnostic Laboratory as H. longicornis before molecular confirmation through PCR amplification of the cox1 gene. Blood samples from infested cattle were collected and assessed molecularly for the presence of T. orientalis, with no pathogenic DNA detected. This report describes the first record of H. longicornis in Oklahoma and the farthest westward detection of this tick in the United States to date.

Characterization of the Complete Mitochondrial Genome of Pleurogenoides japonicus (Digenea, Pleurogenidae): Comparison With the Members of Microphalloidea and Phylogenetic Implications.

Pleurogenoides japonicus (Trematoda: Microphalloidea) is an important parasite in wood frogs with high infection rates and significant ecological, economic, and societal importance. The scarcity of molecular data for these parasites severely limits population genetics and phylogenetic studies. In the present study, for the first time, we determined and described the entire mitochondrial (mt) genome of P. japonicus as the first representative of the family Pleurogenidae. The entire mt genome of P. japonicus was circular, with 15,043 bp (GenBank accession number OR900118), containing 36 genes, comprising 12 protein-coding genes (cox1-3, nad1-6, nad4L, cytb, and atp6), two ribosomal RNA genes, 22 transfer RNA genes, and two non-coding regions. There were 23 intergenic spacers, ranging from 2 to 162 bp, and only one 40 bp overlap between nad4L and nad4 genes in the P. japonicus mt genome. The nucleotide composition of P. japonicus mt genome exhibited a strong AT bias with a 63.75% A + T content, while the AT- and GC-skews were - 0.435 and 0.407, respectively. Comparative analysis demonstrated that the P. japonicus mt genome shared the most common characteristics with Microphalloidea trematodes, and the cox1 gene was the longest and most conserved gene in Microphalloidea trematodes. The gene arrangements of Xiphidiata trematodes were of the same order based on protein-coding genes and rRNA genes, except for tRNA. More than two gene arrangement types exist in Echinostomata and Xiphidiata, and the gene rearrangement events mainly occurred in "trnE-trnG" and "trnG-trnE". Phylogenetic analysis suggested that trematodes of the family Pleurogenidae clustered more with Prosthogonimidae than Eucotylidae. The mt genome data of P. japonicus provide an accurate genetic marker for further studies of Xiphidiata trematodes.