Molecular Identification Methods Research Articles

The Potential of Trichoderma AA1 and AA2 as Local Indonesian Cellulolytic Inoculum in the Fermentation of Gliricidia sepium Leaf Meal

The study aimed to test the cellulolytic ability of Trichoderma AA1 and AA2 inoculum as local Indonesian isolates and their application in the fermentation of Gliricidia sepium leaf meal. Measurement of cellulolytic ability includes qualitative cellulase activity (measurement of clear zone formation around fungal colonies) and quantitative cellulase activity (measurement of the amount of reducing sugar produced by broth enzymes from fungal cultures). Experiments of Gliricidia sepium fermentation using Trichoderma AA1 and AA2 were each designed using Completely Randomized Design. The treatments used consisted of fermentation duration of 0 (control), 2, 4, and 6 days—identification of Trichoderma AA1 and AA2 species using molecular identification method. The research results show that qualitative cellulase activities in Trichoderma AA2 were significantly higher compared to Trichoderma AA1. The CZD/CD ratio of Trichoderma AA1 and AA2 were 2.39±0.11 and 1.97±0.10, respectively. The CMC-ase activity of Trichoderma AA2 was 0.332±0.038 µmol/ml/min, and this was significantly higher compared to TrichodermaAA2, which was only 0.201±0.021 µmol/ml/min. Gliricidia sepium fermentation using AA1 and AA2 inoculum showed increased nutritional value, including soluble protein content, In-vitro digestibility of dry matter, and In-vitro digestibility of organic matter in 2 and 4 days of fermentation. There was a decrease in nutritional value in 6 days of fermentation. Trichoderma AA1 and AA2 were identified as Trichoderma koningiopsis and Trichoderma asperellum, respectively. The study concluded that T. Koningiopsis AA1 and T. Asperellum AA2 were proven to have cellulolytic abilities and could be used as inoculum in Gliricidia sepium fermentation to increase its nutritional value.

Phylogenetic and Morphological Study of Desmodesmus Strains from Can Gio Mangrove Biosphere Reserve

This study focused on five microalgal strains resembling Desmodesmus isolated from the Can Gio Mangrove Biosphere Reserve. The objective was to assess the effectiveness of morphological and molecular methods for algal identification and to evaluate the genetic diversity of the relevant taxa. The five isolated and reference strains were cultured axenically in a BG-11 medium. Both microscopy (at magnifications of 40× and 100×) and molecular techniques (using ITS and 18S rRNA markers) were employed for analysis. Phylogenetic analyses were conducted using Maximum Likelihood and Bayesian inference methods. The results indicated that five of the ten strains were consistently identified using both approaches. Molecular data prompted a taxonomic reassignment for the three remaining strains, while morphological traits were more decisive for two reference strains. Phylogenetic analyses revealed significant genetic diversity within Desmodesmus, highlighting the ecological adaptability of genetically distinct variants. This study emphasizes the reliability of molecular tools in algal taxonomy, particularly for differentiating between Desmodesmus and Scenedesmus-like taxa. It contributes to understanding microalgal genetic diversity in the Can Gio Biosphere Reserve and provides a foundation for future ecological and biotechnological applications.

Molecular and biochemical methods for species identification among the genus Cuscuta

Genus Cuscuta (Convolvulaceae) comprises of stem holoparasitic plants that are scientifically highly valuable, as they cause extensive crop damage. They exhibit high morphological similarity, which complicates their species identification. Determining the taxonomical status of obtained specimens from Bulgaria, is crucial for exploring the genetic diversity of the genus and establishing the phylogenetic relationships within the taxon. Thereby different correlations with geographic region and particular environmental conditions can be established. This study employs molecular and biochemical methods, including ITS region sequence analysis of rDNA, RAPD analysis, and HPLC/MS analysis of flavonoids, to identify Cuscuta species and conduct phylogenetic analysis. Overall, a successful identification has been made - while ITS PCR products provide more accurate species identification and phylogenic information, RAPD profiles are convenient for quick identification. Detailed investigation of the RAPD profiles is required due to differences correlating with the habitats of the species. Nevertheless, flavonoid profiles prove themselves as a valuable biochemical marker for species identification. All of the aforementioned methods are suitable and useful for species identification among the genus Cuscuta, depending on the needs and aims of the conducted analysis.

Molecular and taxonomical study of the genus Chara (Linnaeus 1753) in Israel

Abstract Israel’s climate is characterized by hot and dry summers and cool, rainy winters in the northern and coastal regions, whereas the southern and eastern regions are arid. The aquatic macrophyte Chara is a common genus in Israel, and its distribution varies across Israel’s climatic gradient. Although few studies described the abundance and taxonomy of the local Chara species in Israel, only little is known about the molecular taxonomy of this group in the region. We sampled Chara species along Israel’s climatic gradient and identified specimens using the classical taxonomy key and molecular identification methods. We characterized the molecular taxonomy of four Chara species: C. vulgaris, C. contraria, C. gymnophylla, and C. globata. Notably, the latter species, which was found in a hyper-arid area, was last recorded in Israel ~50 years ago on the northern Mediterranean coast and Judaean Mountains. Chara gymnophylla was more abundant in northern Israel, while C. vulgaris and C. contraria were common in southern Israel. We developed Inter Simple Sequence Repeat markers to identify the four species and sequenced their chloroplast gene rbcL. The phylogenetic trees generated using the rbcL sequences and ISSR-based dendrogram were consistent with the classical taxonomy of Chara.

Molecular detection of zoonotic Spirometra (Cestoda: Diphyllobothriidae) in Javan-spitting cobra (Naja sputatrix) snakes in Indonesia

Abstract. Yudhana A, Kartikasari AM, Edila R, Praja RN, Hamonangan JM, Wardhana AH, Mufasirin, Koesdarto S. 2024. Molecular detection of zoonotic Spirometra (Cestoda: Diphyllobothriidae) in Javan-spitting cobra (Naja sputatrix) snakes in Indonesia. Biodiversitas 25: 4853-4859. Sparganosis, a neglected disease caused by the larvae of Spirometra tapeworms, is considered a serious threat to public health worldwide. However, data on snake prevalence or molecular study of sparganosis in snakes still needs to be improved. In this study, we aim to investigate the prevalence of plerocercoids (spargana) infection in wild-caught Javan-spitting cobra (Naja sputatrix) snakes in Banyuwangi, East Java Province, Indonesia, using morphological and molecular identification methods. A total of 70 Javan-spitting cobra snakes were purchased from local sellers in Banyuwangi, Indonesia. Morphological identification was conducted on a plerocercoids collected from various predilection sites. Moreover, molecular identification was done by polymerase chain the Polymerase Chain reaction (PCR) method and analyzed using the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene. Plerocercoids were identified in 70 snakes, with the prevalence rate recorded at 60%. A total of 231 plerocercoids were collected and divided into 184 (79.65%) in muscular and 47 (20.34%) in subcutaneous tissues. The plerocercoids were macroscopically identified as thin, flat, and white colored with ribbon-like structure and were 2-14 cm long and 2-8 mm wide. Microscopic examination of plerocercoids using the carmine staining method revealed a mouth-like shape anterior side. Furthermore, PCR analysis results reveal that 5 plerocercoid specimens were identified as Spirometra, and each sample shows positive bands at 467 bp. To our knowledge, this study is the first report of Spirometra in species of Indonesian endemic snake primarily on Java Island. These findings constitute a serious potential risk of human sparganosis transmission in Indonesia because wild-caught Javan-spitting cobra snakes are locally used as human food.

Identification of Aspergillus sp. from El-Baida marsh in Algeria: phenotypic and genotypic characterization and industrial enzyme production

The aim of this study was to compare between morphological and molecular identification methods. Two strains (A1, A2) of Aspergillus sp. isolated from El-Baida marsh, were subjected to be identified using morphological characterization, and molecular analysis performed by amplification of the ITS1 and ITS4 regions of rDNA. Morphological analysis indicated that both strains were identified as Aspergillus oryzae. However, by molecular methods A1 strain was identified as Aspergillus alliaceus and A2 strain as Aspergillus oryzae. Our results showed that phenotypic methods were insufficient for correct identification, and the use of genotypic methods is the most reliable. Enzymatic activity of the A2 isolate was evaluated using a plate assay for Lipase, Amylase and Protease production; the results illustrated the capacity of the fungus of the production of the three enzymes, which are mostly used in the production of pharmaceuticals, foods, beverages, and textile. They are found in a vast variety of sources such as animals, plants and microorganisms but microbial enzymes are more stable than the others. In addition, the reality that this fungus is known as a safe microorganism in several domains (food, beverage, cosmetic, and pharmaceutical industries), this opens the research in the future.

Characterization of the sex determining region and development of a molecular sex identification method in a Salangid fish

BackgroundThe short-snout icefish, Neosalanx brevirostris, a member of the Salangidae family, is an economically important fishery species in China. Understanding the mechanisms underlying sex determination in this species has crucial implications for conservation, ecology and evolution. Meanwhile, there is a shortage of rapid and cost-effective genetic methods for sex identification, which poses challenges in identifying the sex of immature individuals in sex determination mechanism studies and aquaculture breeding applications.ResultsBased on whole genome resequencing data, sex-specific loci and regions were found to be concentrated in a region on chromosome 2. All sex-specific loci exhibited excess heterozygosity in females and complete homozygosity in males. This sex determining region contains seven genes, including cytochrome P450 aromatase CYP19B, which is involved in steroidogenesis and is associated with 24 sex-specific loci and two W-deletions. A haploid female-specific sequence was identified as paralogous to a diploid sequence with a significant length difference, making it suitable for rapid and cost-effective genetic sex identification by traditional PCR and agarose gel electrophoresis, which were further validated in 24 females and 24 males with known phenotypic sexes.ConclusionsOur results confirm that N.brevirostris exhibits a female heterogametic sex determination system (ZZ/ZW), with chromosome 2 identified as the putative sex chromosome containing a relatively small sex determining region (~ 48 Kb). The gene CYP19B is proposed as a candidate sex determining gene. Moreover, the development of PCR based method enables genetic sex identification at any developmental stage, thereby facilitating further studies on sex determination mechanisms and advancing aquaculture breeding applications for this species.

Morphological vs. molecular identification of trematode species infecting the edible cockle Cerastoderma edule across Europe

Identifying marine trematode parasites in host tissue can be complicated when there is limited morphological differentiation between species infecting the same host species. This poses a challenge for regular surveys of the parasite communities in species of socio-economic and ecological importance. Our study focused on identifying digenean trematode species infecting the marine bivalve Cerastoderma edule across Europe by comparing morphological and molecular species identification methods. Cockles were sampled from ten locations to observe the trematode parasites under a stereomicroscope (morphological identification) and to isolate individuals for phylogenetic analyses using two gene markers, the small sub-unit ribosomal (18S) RNA gene (SSU rDNA) and the mitochondrial cytochrome c oxidase subunit 1 (cox1). For the first time, we compared both morphological identification and phylogenetic analyses for each of the 13 originally identified species. First, we identified a group of five species for which morphological identification matched molecular results (Bucephalus minimus, Monorchis parvus, Renicola parvicaudatus, Psilostomum brevicolle, Himasthla interrupta). Second, we identified a group of six species for which molecular results revealed either misidentifications or cryptic diversity (Gymnophallus choledochus, Diphterostomum brusinae, Curtuteria arguinae, Himasthla quissetensis, H. elongata, H. continua). Third, our analyses showed that all sequences of two expected species, Gymnophallus minutus and G. fossarum, matched between the two, strongly suggesting that only G. minutus is present in the studied area. Our study clearly demonstrates that molecular tools are necessary to validate the trematode species composition. However, with 17 distinct genetic lineages detected, some of which are not fully identified, future studies are needed to clarify the identity and status (regular vs. accidental infection) of some of these cryptic trematode species.

Phenotypic and Molecular Identification of M.Canis

The current study was conducted to compare morphological and molecular identification methods for the fungus M.canis isolated from 42 patients with dermatophytosis who were clinically diagnosed by a dermatologist, after review by a consultant dermatologist and venereologist in Al-Muthanna Governorate at Al-Hussein Teaching Hospital. In the period from January to November 2024, using traditional methods of isolation and laboratory identification, and molecular methods using polymerase chain reaction (PCR) technology.

Isolation and Molecular Identification of Fungal Contamination on the Eggshells of Lepidochelys olivacea L. Collected from Coastal Waters

Background: One of the causes of the decline in the Olive ridley turtle population is the low egg hatching rate due to contamination by pathogenic fungi. Purpose: This study aims to evaluate the results of the isolation and molecular identification of fungal contamination on the eggshells of olive ridley turtles that failed to hatch from three coastal waters in Bali province. Method: Sampling was done using the purposive sampling technique. Samples of failed hatching eggshells from the coast were collected, as many as 2 eggs/semi-natural nest at TCEC. Samples were cultured on PDA media. Macroscopic and molecular identification methods were carried out to determine the species level of fungal isolates. Results: The results showed that 10 fungal isolates were successfully collected. Isolates KL1 and SG1 had white colony surface characteristics, isolates KL4, SY2 and SY3 showed black colonies, and isolates KL2, KL3, SG2, SG3, SY4 had green colonies. Three similar isolates were sent for further molecular identification. The BLASTn results of the nucleotide sequences of isolates SY2 and SG3 showed a similarity to Aspergillus niger strain PHY105 (91%), while fungal isolate KL1 had a similarity to Fusarium solani strain 87, by 98%. Conclusion: This study provides new information about the pathogenic fungi, Fusarium solani strain 87 and Aspergillus niger strain PHY105, which are suspected to be the cause of the hatching failure of olive ridley turtle eggs.

Non-destructive and rapid identification of yeasts by nearinfrared spectroscopy and machine learning

This study aimed to apply near-infrared (NIR) spectroscopy combined with machine learning techniques to identify yeast strains rapidly and practically, comparing the results with traditional molecular identification methods. Yeasts were isolated from the digestive tracts of aquatic mining insects collected in the extreme north of the Western Amazon (Roraima), Brazil, and preserved through cryopreservation and mineral oil methods. Molecular identification involved PCR amplification and sequencing of ribosomal DNA regions. NIR spectroscopy, coupled with multivariate analysis and machine learning algorithms such as principal component analysis (PCA), hierarchical cluster analysis (HCA), k-nearest neighbor (KNN), and soft independent modeling by class analogy (SIMCA), was used to analyze and classify the yeast samples, accurately identified yeast strains at the genus and species levels, achieving 100% accuracy in both the calibration and validation sets. The results indicate that this method provides a rapid, non-destructive, and environmentally friendly alternative to traditional molecular techniques, making it suitable for real-time, in situ analysis with minimal sample manipulation.

Diagnostic and surveillance testing capability for mpox in the EU/EEA, September 2024.

In response to the increasing number of mpox cases caused by monkeypox virus (MPXV) clade I in the African continent and the first reported travel-related clade Ib case of mpox in EU/EEA, the European Centre for Disease Prevention and Control surveyed national capability for detection and characterisation of MPXV in the EU/EEA. The results showed high level of capability for case confirmation by PCR, alongside molecular typing methods for identification of MPXV clades and/or clade I subclades within the EU/EEA.

Species identification and pollination biology of an economically important true halophyte, Salicornia brachiata Roxb.

Members of the genus Salicornia have gained a global attraction due to their ability to thrive under high saline conditions and as potential candidates in saline agriculture. However, it has been a taxonomically challenging genus for decades since the members show plastic responses to extreme environmental conditions and due to incongruences between morphological and molecular identification methods. While only a handful of commercially grown Salicornia species are fully described, most of the species including S. brachiata, a native species in the Indian sub-continent, Myanmar, and Sri Lanka are poorly described. With the potentials in adapting S. brachiata in saline agriculture, the aim of this study was to establish a morphology and DNA barcode-based species delineation system and to study pollination biology for future crop improvement projects. Tentatively identified S. brachiata plant samples were collected from two populations in Sri Lanka and completely described. GenBank lacked authenticated barcode data for S. brachiata except for one chloroplast genome to which the matK sequence obtained in the present study matched with 100 % identity. For the first time, well defined sequences of three barcode regions, ITS, ETS and matK, of S. brachiata were made available for accurate species identification. Reproductive dynamics in different parts of the inflorescence was studied. A facultative xenogamous mating system was recorded for the first time in the genus and while the lower florets in the cladode showed a preference towards outcrossing, the upper florets displayed adaptations for selfing. Data could be effectively utilized in future Salicornia breeding programs.

Isolation of Lactobacillus spp. from Healthy Infants' Feces and Study of their Antibiotic Susceptibility

This study aimed to isolate Lactobacillus spp. bacteria from the feces of healthy newborns and investigate their susceptibility to antibiotic resistance. 120 fecal samples were obtained from infants under 12 months of age and susceptibility was identified using biochemical tests and molecular identification methods. A total of 30 positive isolates, chosen randomly from the 90 positive samples, were selected to conduct a safety assessment. the isolates were analyzed for their hemolytic activity and susceptibility to antibiotics. Using 16S rRNA sequencing, a total of 30 positive isolates were determined to be lactobacillus spp. The results of the antibiotic resistance test indicated that the majority of the isolates showed susceptibility to tetracycline, chloramphenicol, gentamicin, clindamycin, and amoxicillin while demonstrating resistance to vancomycin, nalidixic acid, streptomycin, and amikacin. conclusion: Based on these specific traits, the isolates of Lactobacillus spp. demonstrated favorable qualities as probiotics.

Isolation and identification of fungi causing post-harvest spoiled mango fruits vended in Hanoi, Vietnam

Mango (Mangifera indica L.) is one of the most popular and nutritious fruits cultivated widely in Vietnam. However, under increasingly harsh climate conditions, mangoes are easily susceptible to fungal invasion and spoiled, thereby reducing mango yield. Therefore, this study was carried out to determine the pathogenic fungal strains of mango to provide useful information for finding effective measures to prevent the diseases. Rotten mango fruits were collected from different markets in Hanoi, Vietnam. Three fungal strains (M1, M2 and M3) were isolated from the studied mangoes samples. All strains were demonstrated as fungal agents associated with mango rot through pathogenicity tests. Microscopic observation showed that the mycelium of these fungal strains was branched and septate. M1 strain formed dark-brown conidiophores and conidia produced on conidiophores. M2 strain produced α- and β-conidia, as well as sub-ovoid and brown sclerotia. Whereas the M3 strain could not produce spores. Additionally, this study determined that all three fungal isolates showed the fastest growth on PDA at 30oC. The optimum growth of the M1 and M3 strains was observed at pH 5.0 while the M2 strain grew actively at pH 7.0 and 8.0. All selected strains showed the ability to produce extracellular enzymes, in which the M1 strain synthesized both cellulase and pectinase while the M2 and M3 strain secreted only pectinase. Finally, by molecular identification method, the isolates (M1, M2 and M3) were identified as Aspergillus niger isolate M1, Phomopsis sp. M2, Lasiodiplodia theobromae M3, respectively.

Mitigating the Threat of Invasive Mosquito Species Expansion: A Comprehensive Entomological Surveillance Study on Kastellorizo, a Remote Greek Island.

The expansion of the tiger mosquito, a vector that can transmit diseases such as dengue, chikungunya, and Zika virus, poses a growing threat to global health. This study focuses on the entomological surveillance of Kastellorizo, a remote Greek island affected by its expansion. This research employs a multifaceted approach, combining KAP survey (knowledge, attitude, practices), mosquito collection using adult traps and human landing catches, and morphological and molecular identification methods. Results from questionnaires reveal community awareness and preparedness gaps, emphasizing the need for targeted education. Mosquito collections confirm the presence of the Aedes albopictus, Aedes cretinus, and Culex pipiens mosquitoes, highlighting the importance of surveillance. This study underscores the significance of community engagement in entomological efforts and proposes a citizen science initiative for sustained monitoring. Overall, this research provides essential insights for developing effective mosquito control programs in remote island settings, thereby emphasizing the importance of adopting a One Health approach to mitigate the spread of vector-borne diseases.

A Microcosm Analysis of Species-Specific Responses of Chironomidae on Heavy Metal Pollution in The Nyanza Gulf of Lake Victoria

The Chironomidae family, known as "non-biting midges" in their adult stage and "bloodworms" in their larval stage, consists of diverse dipteran insects inhabiting various global aquatic environments. Despite extensive global research, data on Chironomidae in the polluted Nyanza Gulf of Lake Victoria, Kenya, is scarce, and molecular identification methods have not been explored. This study aimed to quantify heavy metal concentrations in water, sediment, and insect samples and assess their impact on Chironomid species identified using mitochondrial DNA barcoding of the cytochrome oxidase subunit 1 (COI) gene. Analysis of Variance was used to determine if there were any statistically significant differences in heavy metal concentrations across different sample types or locations along the pollution gradient. Chironomids were collected from Nyanza Gulf, focusing on a pollution gradient. Results showed that concentrations of arsenic (As), lead (Pb), and cadmium (Cd) in insect, water, and sediment samples exceeded standard limits, while mercury (Hg) concentrations were within limits. Significant variations (p ≤ 0.05) in Pb levels were observed in water samples, and heavy metal concentrations in sediment samples varied significantly (p ≤ 0.05), with Pb showing the highest variation (p ≤ 0.0001). Insect samples exhibited significant differences (p ≤ 0.0001) in As and Hg contents. Genetic analysis identified two known species: Chironomus transvaalensis at the heavily polluted Kisumu station and Chironomus pseudothummi at the moderately polluted Kendu Bay and Homa Bay stations. Additionally, a unique Chironomus species was found on Ndere Island, a relatively clean site with restricted human activities. Sequence comparisons indicated proximity to global data but also highlighted the evolutionary significance and uniqueness of the identified species. This study demonstrated the potential use of genetic methods in determining Chironomid species diversity, community structure, and abundance in relation to heavy metal concentration. It suggests that heavy metal pollution may act as a selective pressure, driving the evolution of Chironomid species. The study recommends combining genetic approaches with other pollution sources for a comprehensive understanding of using this species in monitoring pollution

Genetic Sex and Origin Identification Suggests Differential Migration of Male and Female Atlantic Bluefin Tuna (Thunnus thynnus) in the Northeast Atlantic

ABSTRACTKnowledge about sex‐specific difference in life‐history traits—like growth, mortality, or behavior—is of key importance for management and conservation as these parameters are essential for predictive modeling of population sustainability. We applied a newly developed molecular sex identification method, in combination with a SNP (single nucleotide polymorphism) panel for inferring the population of origin, for more than 300 large Atlantic bluefin tuna (ABFT) collected over several years from newly reclaimed feeding grounds in the Northeast Atlantic. The vast majority (95%) of individuals were genetically assigned to the eastern Atlantic population, which migrates between spawning grounds in the Mediterranean and feeding grounds in the Northeast Atlantic. We found a consistent pattern of a male bias among the eastern Atlantic individuals, with a 4‐year mean of 63% males (59%–65%). Males were most prominent within the smallest (< 230 cm) and largest (> 250 cm) length classes, while the sex ratio was close to 1:1 for intermediate sizes (230–250 cm). The results from this new, widely applicable, and noninvasive approach suggests differential occupancy or migration timing of ABFT males and females, which cannot be explained alone by sex‐specific differences in growth. Our findings are corroborated by previous traditional studies of sex ratios in dead ABFT from the Atlantic, the Mediterranean, and the Gulf of Mexico. In concert with observed differences in growth and mortality rates between the sexes, these findings should be recognized in order to sustainably manage the resource, maintain productivity, and conserve diversity within the species.

Fish eggs in the diet of White Sea cod Gadus morhua marisalbi (Gadiformes: Gadidae): species identification using DNA-based methods

Trophic relationships reflect the peculiarities of the functioning of communities of aquatic organisms and analysis of their structure and dynamics can be used to improve fisheries management and assess the state of marine ecosystems. Populations at the margins of the species' range have particular adaptations, including feeding habits, to extreme conditions, which makes them a convenient model object for studying the possible consequences of ecosystem changes under the influence of global factors. The cod Gadus morhua marisalbi Derjugin, 1920 permanently inhabits the coastal waters of the subarctic White Sea, located at the northeastern margin of the geographic range of the Atlantic cod G. morhua Linnaeus, 1758. This coastal cod population strongly depends on the local environmental conditions and available food resources during the summer feeding period. In addition to the main food organisms (fish, polychaetes, crustaceans), the eggs of fish spawning in shallow coastal areas in summer are also important in the diet of cod at this time. According to published data, fish eggs found in significant quantities and most often in the stomachs of cod belonged to threespine stickleback and lumpfish, but this has not been confirmed by any experimental research. Besides, the possible presence of herring eggs in the diet of cod remains unproven due to the difficulties of its visual identification. In this work, we used molecular genetic methods for species identification of fish eggs found in cod stomachs. At the first stage, the species identification of DNA isolated from eggs was done using two sets of species-specific primers: the first which we designed for the mitochondrial COI gene of stickleback, cod, and herring, and the second, published for a microsatellite locus of herring. At the second stage, the species identity of DNA extracted from fish eggs was checked using DNA barcoding. The results showed that cod stomachs contained herring and stickleback eggs, and in a number of cases we found the co-occurrence of eggs of these species. Of the 29 DNA preparations studied, fourteen were successfully amplified with microsatellite primers for herring, and thirteen amplified with primers for stickleback. Interannual differences in the occurrence of herring eggs in samples were insignificant, while the frequency of occurrence of stickleback eggs in 2018 was higher than in 2017. Our analysis did not reveal significant intra-annual differences between the observed and theoretically expected number of cases of simultaneous occurrences of herring and stickleback eggs. The high frequency of occurrence of herring and stickleback eggs in cod stomachs revealed in this study indicates the importance of these components in the diet of cod and provides new knowledge on the trophic role of cod as a key species in the White Sea ecosystem. Analysis of available data on the spatial and temporal diet patterns of coastal White Sea cod shows that fish eggs can significantly contribute to the total amount of food consumed by this species during the summer.

Deciphering the Plastome and Molecular Identities of Six Medicinal "Doukou" Species.

The genus Amomum includes over 111 species, 6 of which are widely utilized as medicinal plants and have already undergone taxonomic revision. Due to their morphological similarities, the presence of counterfeit and substandard products remains a challenge. Accurate plant identification is, therefore, essential to address these issues. This study utilized 11 newly sequenced samples and extensive NCBI data to perform molecular identification of the six medicinal "Doukou" species. The plastomes of these species exhibited a typical quadripartite structure with a conserved gene content. However, independent variation shifts of the SC/IR boundaries existed between and within species. The comprehensive set of genetic sequences, including ITS, ITS1, ITS2, complete plastomes, matK, rbcL, psbA-trnH, and ycf1, showed varying discrimination of the six "Doukou" species based on both distance and phylogenetic tree methods. Among these, the ITS, ITS1, and complete plastome sequences demonstrated the highest identification success rate (3/6), followed by ycf1 (2/6), and then ITS2, matK, and psbA-trnH (1/6). In contrast, rbcL failed to identify any species. This research established a basis for a reliable molecular identification method for medicinal "Doukou" plants to protect wild plant resources, promote the sustainable use of medicinal plants, and restrict the exploitation of these resources.