Taxonomic Identification Research Articles

Karyotyping: Basics and Application in Forestry

Karyotyping is the method of assessing an organism's chromosomal number and structure, is essential for comprehending the genetic composition of species, especially those used in forestry. This method provides important insights into the genetic diversity, evolution, breeding, and conservation of trees and other forest species by examining their chromosomal features. Karyotypes explain how many chromosomes each organism has and how they appear under a light microscope. Length of the chromosome, centromere location, banding pattern, sex-chromosome variations, and other physical traits can also be determined by karyotyping. At traditional level, by using light microscope, we can study chromosomes but to generate chromosomal data, sophisticated methods like fluorescence in-situ hybridization (FISH) and genomic in-situ hybridization (GISH) can be employed. From taxonomical classification to identify different types of abnormalities, karyotyping is important. There is a lack of study related to karyotyping in forestry species. This study summarizes the basics of karyotyping and the work related to it, has been done in forestry species. This review article deals with the importance of karyotyping such as how they contribute to the conservation biology of the forestry species, how it helps in the development of taxonomical identification marker and identification in different types of chromosomal abnormalities. It also covers how karyotyping can be done. From traditional methods i.e use of microscopes to the merge of different recent developments in the area of karyotyping helps not only in enrichment of cytogenetics data but also substitute the genomics and transcriptomics in the area of genetic diversity studies. Basics of karyotyping, different types of banding techniques, protocols of some of the techniques, its methodological developments, and its various applications in forestry have also been discussed in this review. Application of karyotyping in different forestry species is limited. With the advancement of imaging techniques and other genetic tools, biodiversity and forests can be managed sustainably.

MARLOWE: An Untargeted Proteomics, Statistical Approach to Taxonomic Classification for Forensics.

General proteomics research for fundamental science typically addresses laboratory- or patient-derived samples of known origin and composition. However, in a few research areas, such as environmental proteomics, clinical identification of infectious organisms, archeology, art/cultural history, and forensics, attributing the origin of a protein-containing sample to the organisms that produced it is a central focus. A small number of groups have approached this problem and developed software tools for taxonomic characterization and/or identification using bottom-up proteomics. Most such tools identify peptides via database search, and many rely on organism-specific peptides as markers. Our group recently introduced MARLOWE, a software tool for taxonomic characterization of unknown samples based on de novo peptide identification and signal-erosion-resistant strong peptides, which are shared peptides distributed in a taxonomy-dependent manner. In the current work, we further characterize the utility of MARLOWE using publicly available proteomics data from forensically-relevant samples. MARLOWE characterizes samples based on their protein profile, and returns ranked organism lists of potential contributors and taxonomic scores based on shared strong peptides between organisms. Overall, the correct characterization rate ranges between 44 and 100%, depending on the sample type and data acquisition parameters (with lower numbers associated with lower-quality data sets). MARLOWE demonstrates successful characterization of true contributors and close relatives, and provides sufficient specificity to distinguish certain microbial species. MARLOWE demonstrates its ability to provide insight into potential taxonomic sources for a wide range of sample types without prior assumptions about sample contents. This approach can find utility in forensic science and also broadly in bioanalytical applications that utilize proteomics approaches for taxonomic characterization.

Shape and Size Variations in the Astragalus of Large and Small Bovids

The astragalus acts as a crucial intermediary in the tarsal joint, facilitating the transfer of body weight to the lower extremities while also ensuring both stability and mobility through its articulation with adjacent bones, enabling efficient movement. This study investigates the morphological diversity of the astragalus in three modern cattle breeds, one goat breed, and three sheep breeds using three-dimensional geometric morphometric methods. A total of 225 specimens were analyzed in this study. Large bovids, such as the Holstein and Simmental cattle breeds, had a more developed lateral edge of the proximal trochlea and a more rounded astragalus. In small bovids, the Akkaraman and Morkaraman sheep breeds displayed a more prominent medial edge and a rounded astragalus form, while the Hair goat breed exhibited a rectangular structure with a well-developed lateral projection. The results of this study demonstrate that the astragalus serves as a reliable morphological marker for differentiating between large and small bovids, as well as between closely related taxa such as sheep and goats. These shape variations, particularly observed in the proximal trochlea, reflect functional adaptations to size, body mass, and locomotor demands, making the astragalus a critical element for taxonomic identification and biomechanical analysis.

First assessment of biofouling assemblages in the northern Red Sea, an important region for marine non-indigenous species transfer

IntroductionMaritime traffic and coastal urbanisation significantly contribute to the introduction and proliferation of non-indigenous species (NIS). However, the lack of information might prevent effective monitoring in data-limited regions, particularly in areas experiencing demographic growth, where monitoring biofouling communities could offer crucial insights into the dynamics of NIS invasions. This study represents a baseline characterization of the biofouling communities in the northern Saudi Arabian Red Sea (NEOM region) prior to extensive urban, industrial, and commercial development.MethodsSamples were collected in November 2023 and February 2024 from seven sites within the region. At each site and time, three settlement structures, each with a PVC panel attached to a brick and rope, were sampled after 3-months deployment. Panels were analysed with photo quadrat analysis (PhQd), examined to manually collect sessile macroinvertebrate specimens for taxonomic identification using morphological and DNA barcoding analysis, and scraped for bulk DNA analyses using DNA metabarcoding. Five water samples were also collected for environmental DNA (eDNA) analysis. The environmental characteristics of each site were obtained by deploying temperature data loggers and through an environmental risk assessment.ResultsCommunity patterns observed from the PhQd and bulk DNA datasets revealed a clear separation of two sites within Sharma lagoon from the remaining sites. The eDNA analysis of seawater confirmed these spatial differences, also detecting a variation between sampling times that was not observed with the other approaches. A total of 20 NIS and 18 cryptogenic species were recorded, from which 12 were identified morphologically and the remaining detected with molecular methods only. The generally low NIS coverage confirms that the NEOM region is less affected by marine biological invasions compared to other anthropized habitats within the Red Sea. However, sites in the Sharma lagoon showed high human pressure levels and comparatively higher coverage of cryptogenic bryozoans on panels.DiscussionThis study provides the first detailed assessment of biofouling communities in NEOM, establishing a baseline and contributing to a regional species reference library for non-indigenous and cryptogenic species. As coastal development spreads, it presents both challenges and opportunities, highlighting the need for sustainable, ecosystem-based approaches to protect valuable natural areas. This baseline is essential for future monitoring of biofouling dynamics as the region develops.

Safety evaluation of the food enzyme acylglycerol lipase from the genetically modified Penicillium sp. strain AE‐LGS

Abstract The food enzyme acylglycerol lipase (glycerol‐ester acylhydrolase, EC 3.1.1.23) is produced with the genetically modified Penicillium sp. strain AE‐LGS by Amano Enzyme Inc. The Panel could not conclude on the taxonomic identification of the production strain due to a lack of appropriate data. The genetic modifications do not give rise to safety concerns. The food enzyme was free from viable cells of the production organism and its DNA. The food enzyme is intended to be used in two food manufacturing processes. The compositional data provided were insufficient to characterise the food enzyme batches for commercialisation and the batch used for toxicological testing. As a result, the suitability of the batch used in the toxicological tests could not be established. The toxicological studies were thus not considered and the dietary exposure not calculated. A search for the homology of the amino acid sequence of the acylglycerol lipase to known allergens was made and no match was found. The Panel considered that a risk of allergic reactions upon dietary exposure to the food enzyme cannot be excluded, but the likelihood is low. Taken together, due to insufficient data regarding the production strain and the food enzyme composition, the Panel could not complete the safety assessment of the food enzyme.

VIMP approach to fern’s sporophyte/gametophyte stages: taxonomic and phylogenetic electrochemical correlations

Abstract The application of the voltammetry of immobilized particles methodology (VIMP) to microparticulate films prepared from the ethanolic extracts of different fern species in contact with aqueous acetate buffer is studied. The observed voltammetric responses can be mainly attributed to the oxidation of polyphenolic components of fern tissues. These responses provide a basis for linking the gametophyte stage to the fern sporophyte, facilitating taxonomic identification during the early developmental phase when distinguishing morphological characters are not clearly discernible. Data collected for 10 species from the Davalliaceae, Aspleniaceae, Athyriaceae, Polypodiaceae, Dryopteridaceae, Pteridaceae, and Osmundaceae families permit an electrochemical grouping consistent with taxonomic and phylogenetic data derived from available molecular genetic analysis.

Diversity and Mitochondrial CO1-Based Barcoding of Orthopteran Species of Bangladesh

Recognizing the ecological and economic significance of Orthoptera species in Bangladesh, maintaining an up-to-date inventory of the species within the country becomes crucial. Building upon previous records, the current investigation places a central focus on documenting the taxonomic identity, species distribution, and evolutionary relationships of Orthoptera species in Bangladesh, employing the mitochondrial COI-based barcoding technique. The findings reveal a total of 13 orthopteran species across 13 genera and 9 families, with three species identified through barcoding. Notably, Phlaeoba sikkimensis is recorded as a first for the Bangladeshi entomofauna, highlighting its importance in local biodiversity. The comprehensive genetic data presented in this study significantly enhance our understanding of orthopteran diversity, facilitating the development of effective measures for their conservation and sustainable management, thus fostering a harmonious coexistence with these ecologically and economically valuable species. J. Asiat. Soc. Bangladesh, Sci. 50(1-2): 125-136, June-December 2024

Chemotaxonomy and Bioactive Potential of High-Mountain Plantago atrata.

Plantago atrata Hoppe is a high-altitude mountain plant exposed to harsh environmental factors. This study aims to elucidate the ecological, phytochemical and pharmacological characteristics of this lesser-known plantain. Despite nutrient-poor peat soil, the leaves of P. atrata contained increased levels of phosphorus, potassium and calcium, and the anatomy revealed an isobilateral mesophyll. Molecular taxonomic identification and phylogenetic analyses confirmed the classification of the studied plant as P. atrata and its clustering with narrow-leaved plantains growing in extreme conditions. Detailed phytochemical profiles revealed primary and secondary metabolites that can be used as taxonomic identifiers and for bioactivity studies. The methanol fraction enriched in phenolic compounds appeared biologically active - by displaying limited antimicrobial activity, however, it possessed significant radical-scavenging and anti-inflammatory capacity. The data demonstrate that P. atrata's leaf parameters and phytochemical arsenal assure plant survival and reveal the plantain's pharmaceutical potential in inhibiting inflammation via the classical pathway of Complement activation.

First Report of Pantoea ananatis Causing Bacterial Leaf and Panicle Blight of Rice in Louisiana, USA.

In July 2023, panicle and leaf blight-like symptoms were observed from the rice (Oryza sativa) variety, PVL03, in research field plots in Louisiana (Rayne, LA 70578, USA; 30.21330⁰ N, 92.37309⁰ W). We observed that chlorosis started on the tips and spread throughout the whole leaf. We also observed panicle blight-like symptoms (grain discoloration of rice hulls) on infected rice plants at the heading stage, which ranged from dark brown to black. Disease incidences were approximately 70% in the observed plots. Five plants were sampled from five plots (one plant per plot). Leaf tissues including both symptomatic and healthy parts (~ 1 cm2) were surfaced sterilized and placed on CCNT medium, a semi-selective medium for the rice bacterial panicle blight pathogen Burkholderia glumae (Kawaradani et al., 2000), and on Luria-Bertani (LB) agar supplemented with cycloheximide (50 µg/ml) for non-selective isolation of bacteria. For the panicle samples, the surfaced sterilized rice kernels were dehulled and ground in sterile distilled H2O using a sterile mortar and pestle. The homogenate was spread on the CCNT media and LB agar supplemented with nitrofurantoin (50 µg/ml) and cycloheximide (50 µg/ml). After ~72 h at 41⁰C, round, smooth and yellow-colored bacterial colonies were observed on both media. The two isolates, one from an infected rice leaf (LPL-1) and the other from the infected rice panicles (LPP-1), were initially identified as Pantoea ananatis based on PCR-amplified 16S rDNA sequences (Weisburg et al., 1991). The taxonomic identity of these two isolates was further confirmed through whole genome sequencing (NCBI Accession ID: PRJNA1092676), which was performed through a hybrid sequencing approach using Oxford Nanopore sequencer & Illumina NextSeq2000 system (Plasmidsaurus Inc., South San Francisco, CA, USA). To satisfy Koch's postulates, we inoculated six one-month-old rice plants (cv. Kitaake) with LPL-1 and LPP-1, respectively, by pricking the stems with a sterile toothpick touched to the overnight grown bacterial culture on LB agar (~ 5-8 x 105 bacterial cells). This virulence assay was conducted twice, and rice plants inoculated with sterile distilled water were included as the negative control. By 27 days post inoculation (DPI), the inoculated plants developed symptoms on leaves and panicles, which were like the symptoms originally observed in the field, and additional symptoms such as shot holes and stem necrosis were also observed. From leaf and panicle samples of the artificially inoculated plants, we reisolated bacteria and confirmed the identity of the bacteria using the P. ananatis specific PCR primers PANA_1080 61F and PANA_1080 1009R (Asselin et al., 2016). To the best of our knowledge, this is the first report of bacterial panicle and leaf blight of rice caused by P. ananatis in Louisiana. Since P. ananatis is a recently emerging pathogen of rice worldwide, including the United States (Yu et. al., 2022; Lu et al., 2022; Luna et al., 2023), it is crucial to monitor the occurrence of this pathogen in other rice-growing areas as a potential threat to US rice production.

Adaptive sequence alignment for metagenomic data analysis.

With advances in sequencing technologies, the use of high-throughput sequencing to characterize microbial communities is becoming increasingly feasible. However, metagenomic assembly poses computational challenges in reconstructing genes and organisms from complex samples. To address this issue, we introduce a new concept called Adaptive Sequence Alignment (ASA) for analyzing metagenomic DNA sequence data. By iteratively adapting a set of partial alignments of reference sequences to match the sample data, the approach can be applied in multiple scenarios, from taxonomic identification to assembly of target regions of interest. To demonstrate the benefits of ASA, we present two application scenarios and compare the results with state-of-the-art methods conventionally used for the same tasks. In the first, ASA accurately detected microorganisms from a sequenced metagenomic sample with a known composition. The second illustrated the utility of ASA in assembling target genetic regions of the microorganisms. An example implementation of the ASA concept is available at https://github.com/elolab/ASA.

Multi-Stage Screening Purification: A New and Effective Method for Cleaning Diatom Samples from Marine Sediments

Diatoms are critical indicators in marine paleoecology and contemporary ecosystem studies, yet the accuracy of diatom analysis depends on effective purification methods. Current screening and purification techniques often yield low accuracy. This study introduces a multi-stage screening purification method that integrates both physical and chemical techniques to enhance the extraction of diatom remains from marine sediments, thereby improving the reliability of subsequent analysis. Using surface sediment samples from the Pacific Ocean, we compared the effectiveness of three purification methods: heavy liquid suspension purification, single-layer screening purification, and the newly developed multi-stage screening purification method. The study aimed to evaluate the impact of each method on diatom abundance, valve size distribution, and the accuracy of taxonomic identification. The results revealed that the multi-stage method significantly improved the accuracy of diatom abundance calculation. With this method, diatom abundance was 21.9 times higher than that obtained using the heavy liquid suspension method and 6.5 times higher than that obtained using the single-stage method. This method also proved to be cost-effective, easy to use, and produced samples with low impurity levels, which enhanced microscopic observation, identification, and the reliability of taxonomic statistics. Key factors influencing diatom abundance included sieve pore size, heavy liquid concentration, and valve size and density, while the type of acid and oxidant had minimal effect. Additionally, the multi-stage screening method facilitated the classification of diatoms into various size categories, providing a comprehensive view of diatom size distribution, including nano-sized diatoms (diameter < 20 μm) that are often overlooked in traditional studies. These findings demonstrate that the multi-stage screening purification method is an effective tool for improving the analysis of diatom remains in sediments, potentially refining the accuracy of diatom-based environmental investigations and paleoceanographic reconstructions.

Reviving diversity: cryoprotectants and culturing methods enhance recovery of mammalian gut microbes from field samples

Abstract The field of microbial ecology is increasingly recognizing the need for methods to isolate and culture gut microbes to better understand how these microorganisms impact animal physiology, especially in mammalian hosts. Currently, there is a lack of clear methods to store microbial samples for cultivability, especially when samples are collected from the field, transported to the laboratory, and preserved under long-term storage for weeks to months compared to mere days in the biomedical field. Here, the cecal contents of groundhogs (Marmota monax) were processed and stored with or without various preservation solutions at −80 °C for at least 2 months. All microbial samples were then grown in distinct nutrient media in liquid and plate conditions and were incubated under anaerobic and aerobic environments. Treatment comparisons revealed that the samples stored in preservation solutions containing 1 or more cryoprotectants provided the greatest and most consistent bacterial densities. To test the long-term storage efficacy of the preservation solutions, we inventoried taxonomic identities and abundances of these cultures using 16S rRNA amplicon sequencing. Our findings highlight that: (1) preserved samples containing cryoprotectants exhibited the highest microbial richness and diversity and resembled the original cecal samples the most when grown under anaerobic conditions; and (2) the effect of individual animal identity was detectable in the membership of cultured communities, irrespective of preservation solutions. Our study is the first to demonstrate the importance of preservation solutions containing multiple cryoprotectants for long-term storage and further microbial culturing and novel isolation. Understanding and improving storage methods that preserve microbial physiology and conserve their compositional diversity is essential for field-collected samples useful in mammalian microbiome and culturomics studies, promoting a better comprehension of the identity and function of wild host-associated microbiomes.

First Report of Bacterial Wilt of Ginger Caused by Ralstonia pseudosolanacearum in the Continental United States.

Ginger (Zingiber officinale) is an herbaceous perennial in the Zingiberaceae family grown primarily in tropical to subtropical biomes as a culinary spice, a traditional medicine, and a landscaping plant. While ginger grows at soil temperatures above 20°C, several farmers in the upper Midwestern US farmers grows short-season ginger in high tunnels. In 2023 and 2024, growers in southeastern Minnesota reported a new disease of ginger. USDA traced the origin of the ginger rhizomes imported by these growers to Peru (Soto-Heredia et al., 2024). Plants exhibited wilting, chlorosis, pseudostem collapse, and plant death. Rhizomes were squishy with a fetid, rotting odor. Disease incidence ranged from scattered plants to 50% of the crop. Symptomatic samples tested positive for Ralstonia solanacearum species complex (RSSC) using AgDia ImmunoStrips. Rhizome tissue extracts plated onto CPG + 1% tetrazolium chloride agar and mSMSA agar produced irregular, round-to-fluidal white colonies with pink centers after 48 h of incubation at 28°C (Kelman 1954; Engelbrecht 1994). Diagnostic RSSC phylotype-specific multiplex PCR primers Nmult21:1F, Nmult21:2F, Nmult23:AF, Nmult22:InF, Nmult22:RR, and 759f/760r confirmed isolate identity; two isolates (UMN24-1, UMN24-2) yielded bands at 280 bp and 144 bp, identifying them as RSSC and phylotype I respectively (Fegan and Prior 2005; Opina et al. 1997). Additionally, 16S rRNA PCR of these isolates was sequenced using primers 27F (5'-AGAGTTTGATCMTGGCTCAG-3')/1492R (5'-TACGGYTACCTTGTTACGACTT-3') and shared >99% identity (isolate UMN24-1: 99.00% 27F, 99.53% 1492R; UMN24-2: 99.32% 1492R) with R. pseudosolanacearum (Rps) strain Gj707 (GenBank CP104497.1). Eight ginger plants in individual pots were grown in a Conviron growth chamber (28°C, 250 μmol/m2/s, 16 h photoperiod) and inoculated at the six-leaf stage (6 weeks old) by pipetting 109 CFU of the isolate, UMN24-1, suspended in sterile water into a 30°angled 1 cm incision on the pseudostem. We scored disease incidence based on wilted leaves per total leaves on each plant for 14 days post inoculation (dpi). While control plants remained healthy, inoculated plants showed symptoms (wilting, vascular discoloration, chlorosis) at 4 dpi, were 100% wilted by 6 dpi, and all pseudostems completely collapsed by 11 dpi (Fig. S1). Bacteria recovered from the inoculated plants matched the original isolate in appearance and positively identified as RSSC phylotype I using the PCR analyses described above. To confirm taxonomic identification of UMN24-1, the complete genome of this isolate was sequenced using hybrid Illumina and Oxford Nanopore sequencing (NCBI BioProject ID PRJNA1178296, release date upon publication) (Fig. S2). Average nucleotide identity (ANI) analysis of UMN24-1 using KBase's FastANI app reported 99.8% identity shared with Japanese RSSC phylotype I strain MAFF311693 (GenBank GCF 015698385.1). We also sequenced UMN24-2, and its phylogenetic analysis revealed that the two isolates are clonal (Fig. S2). Hence, we did not submit its sequence to the NCBI. This marks the first documented case of Rps in ginger in the United States. Similar to bacterial wilt in ginger closely related crop like turmeric has been repeatedly reported in northern Europe in recent years (EPPO 2024). While RSSC phylotype I is geographically limited by its minimal cold tolerance, the emergence of a pathogen with a potentially broad host range has worrying implications in a changing climate.

Toward an Efficient Differentiation of Two Diaporthe Strains Through Mass Spectrometry for Fungal Biotyping.

Considering that fungi display a great morphological, ecological, metabolic, and phylogenetic diversity, their taxonomic identification is extremely important because it helps us establish important information about each species and its possible biochemical and ecological roles. Traditionally, the identification of fungi at the species level has been carried out with molecular tools such as DNA sequencing, but it still represents a huge challenge today due to the heterogeneity of the fungal kingdom, making the task of identification a complex and difficult process. Biotyping, a type of chemotaxonomy, has been developed in the field of the identification/differentiation and classification of micro-fungi through tools such as mass spectrometry (MS). Here, two endophytic strains isolated from two different hosts were cultivated and studied regarding their morphology and molecular biology. Morphology analysis determined the strains as Diaporthe, and the molecular analysis results grouped them as D. melongenae. We sought a faster and less complex way of differentiating these fungal strains of interest through an MS chemical profile and MS/MS data using a low-resolution mass spectrometer. Additionally, we linked this information with the structure of compounds previously isolated in the genus Diaporthe. Studies conducted using this technique allowed us to propose the structure of distinctive molecules that are unique to each strain and share compounds common to this genus (13 compounds in total). In addition, this is the first report of secondary metabolites in D. melongenae. The dataset demonstrates that the two strains under investigation can be distinguished via mass spectrometry, suggesting host affinity; both exhibits pronounced differences in their chemical profiles across all culture media and incubation periods with the parameters described herein.

Assessments of the earliest bats from the Quaternary of Serra da Mesa (Goiás, Brazil): phylogenetic insights and biogeographic modelling on the new extinct species of Rhinophylla, the first fossil record of the subfamily Rhinophyllinae (Chiroptera, Mammalia)

ABSTRACT Recovered from Quaternary deposits in Carneiro Cave (Serra da Mesa, Brazil), associated with a clayey-sandy sedimentary influx older than the overlying calcite layer, which has been dated to 182.8 ± 1.2 ka, a new extinct species of Rhinophylla is described based on a proximal radio fragment. This discovery represents one of the three oldest occurrences of Chiroptera in the exposed stratigraphic profile. The fossil record of Chiroptera consists of 1,117 bone fragments, distributed across Pleistocene and Holocene sedimentary packages. The chronology of this sedimentary sequence is established using dated layers, all of which are fully consistent with their order of deposition. A comparative study of the radioulnar morphology was conducted to determine the taxonomic identity of the new fossil bat. A strict consensus tree for phyllostomids was obtained, surprisingly consistent with recent molecular phylogenies, unequivocally places the new fossil in the subfamily Rhinophyllinae. Biogeographic modelling of Rhinophylla, typically characterised by an ecomorphology of small specialised epiphytic frugivores, suggests that around 200 ka (the fossil’s age reference) the genus may have had a broader distribution across Central Brazil than is observed today. This is consistent with the fact that Rhinophylla is currently absent in the surrounding region of the Carneiro Cave.

Early development of vertebral column and appendicular skeleton in Naozhou Larimichthys crocea (Richardson, 1846).

Understanding the developmental sequence characteristics of the vertebral and appendicular skeletons of the larvae and juveniles of Larimichthys crocea (Naozhou population) can provide theoretical basis for seedling cultivation, environmental adaptation, and taxonomic identification. The cartilage-bone double staining method was used to stain, observe, and analyse the vertebrae, pectoral fins, anal fins, caudal fins, and dorsal fins of the larvae and juveniles of L. crocea (0-30 days post-hatching [DPH]). Results showed that the notochord of the larvae and juveniles of L. crocea was tubular. At 6 DPH, there was obvious segmentation. At 8 DPH, the neural arches began to differentiate, and at 10 DPH, the haemal arches began to differentiate, with complete segmentation of the notochord. At 14 DPH, the dorsal and ventral ribs became clear, and the neural and haemal spines were completely formed by the elongation of the neural and haemal arches, respectively. At 18 DPH, the vertebral bones began to ossify, and ossification was complete at 28 DPH. The median fins of the larvae and juveniles of L. crocea formed in the order of caudal fin, anal fin, and dorsal fin. Among the 800 L. crocea larvae samples, 248 were observed to have skeletal deformities, with a deformity rate of 31.00%. The spine of L. crocea consists of 26 vertebrae, with developmental abnormalities mainly including vertebral anterior convexity, bifurcation of neural spines, vertebral body fusion, and redundancy of neural spines. The above results provide a theoretical basis for enriching the developmental biology of L. crocea.

Fatty Acid Content and Profile in Ulva lactuca in Response to Exposure to Variable Growth Conditions in Indoor Photobioreactors.

Seaweed presents a sustainable alternative source of valuable fatty acids (FAs) involving omega-3 (n-3) and omega-6 (n-6). As such, there is great potential to reduce pressure on wild fish populations, helping to combat overfishing and its associated global impacts. This study explored the effect of various environmental factors on the FA content and profile of Ulva lactuca using indoor photobioreactors. The taxonomic identity of U. lactuca was confirmed through DNA sequencing using 3 markers (rbcL, ITS, and tufa). The effects of temperature (8, 20, and 30 °C), seawater salinity (3.5, 3.0, 2.5, and 2.0% w/v), nutrient type and concentration (0 or 6.4 ppm, consisting of 50% w/w N-NO3, 50% w/w N-NH4, and 0-1 ppm P-PO4), and irradiance (50, 100, and 150 μmol photons m-2 s-1) were evaluated. This study assessed their influence on U. lactuca's biomass production rate (BPR), dry weight (DW), ash content (AC), and FA composition after 7 and 21 days. The results revealed that after 21 days, the polyunsaturated FA (PUFA) content decreased with the increasing seawater salinity (i.e., 38.9% ± 0.7, 33.8% ± 0.4, and 27.0% ± 0.4, and 6.6% ± 0.1 for a salinity of 2.0, 2.5, 3.0, and 3.5% w/v, respectively). The content of n-3 after 21 days increased significantly under the following conditions: 8 °C, a salinity of 2.5% w/v, 6.4 ppm of nitrogen without the addition of phosphorous, and an irradiation of 50 and 150 μmol photons m-2 s-1, affording a low n-6/n-3 proportion that fits a desirable level of an n6/n3 ratio (1-10) for a balanced nutritional diet.

New Record of Encarsia protransvena and Confirmed Occurrence of Encarsia hispida (Hymenoptera: Aphelinidae) as Parasitoids of Singhiella simplex (Hemiptera: Aleyrodidae) in Italy.

Encarsia protransvena (Hymenoptera: Aphelinidae) is recorded here for the first time in Italy as a parasitoid of the whitefly Singhiella simplex (Hemiptera: Aleyrodidae), one of the most invasive alien pests of exotic Ficus species. Singhiella simplex, originating from the Oriental region, has established a global presence. Monitoring of the whitefly and its parasitoids was conducted in the southern areas of Italy, providing crucial insights into their distribution and interactions. The taxonomic identity of E. protransvena, was confirmed by scrutiny of morphological and molecular taxonomic characters. At the time of writing, we also obtained some specimens of Encarsia hispida from the host collected in Campania. We reserve the right to provide bio-ethological information on the species in the future. Comprehensive illustrations and diagnostic features are provided for the host and the parasitoids. An identification key is included for all Encarsia species associated with S. simplex, which provides a valuable tool to distinguish these aphelinid wasps for future research and applications in biological control programs.

Taxonomic identification and prevalence of hard ticks on different breeds of sheep from district Quetta in Balochistan, Pakistan

Ticks, being ectoparasites, are vectors for the transmission of various pathogens that can infect animals and can also pose a significant threat to human health. Due to hot and humid climatic conditions across different agro-ecological regions of Pakistan, ticks are widespread and infest a diverse range of animal species. This study aimed for taxonomic identification and prevalence determination of ticks in sheep collected from four different marketplaces in the Quetta district of Balochistan. A total of 200 sheep from six different breeds, Balochi, Harnai, Turki, Mari wal, black Shinwari-Afghani, and Kooka, were examined, and ticks were collected from their five distinct anatomical regions. The species are identified over petri-dish and under microscope using their morphological features. Out of total 200 sheep, 107 were identified to be infested by ticks with an overall prevalence of 53.50 %. The class of ≤6 month's age are more prevalent and more prone to tick infestation, with tick prevalence of 55 %. The female sheep were found more prone to tick infestation (56 %) as compared to male species (51 %). A total of 559 species were documented including H. anatolicum, R. microplus, H. rufipes, H. dromedary, R. (Boophilus) annulatus, R. decoloratus, Ixodes Pilosus, R. camicasi, R. pravus, and R. turanicum. R. decoloratus was noted in higher abundance (N; 31) at larvae stage, while H. anatolicum are more in nymph stage (N: 30); the R. (Boophilus) annulatus species are dominating the adult sage (N: 60). The amount of tick infestation also varied according to the season and month of the year: in the summer, the highest number of sheep become infected with ticks, with July having the highest frequency of tick predominance; in the winter, the lowest prevalence of ticks is found in January. The specie abundance was noted to be 559, Specie richness to be 10, and the diversity was calculated as 0.93.

WHO IS WHO IN THE FORAMINIFERAL WORLD: TAXONOMIC REVISION OF SELECTED CIBICIDOIDES AND PLANULINA SPECIES IN RECENT SOUTHEAST PACIFIC MARINE SEDIMENTS

ABSTRACT Cibicidoides and Planulina are common genera of benthic foraminifera used for paleoceanographic reconstructions through the study of their stable isotopes (e.g., δ18O, δ13C) or morphological features (e.g., pore patterns). Often with an epifaunal mode of life, species of these genera are recognized as reliable indicators for assessing bottom water conditions. Hence, precision in their taxonomic identification is necessary for their effective utilization in paleoceanographic studies. To establish criteria for a consistent classification system of selected recent Cibicidoides and Planulina species, stereomicroscope and SEM imaging, manual illustration, and observations were conducted for specimens from surface sediment samples at the continental margin of Chile and Peru (12–44°S) in the Southeast Pacific, one of the most understudied regions worldwide. In the study area, Cibicidoides wuellerstorfi specimens showed morphological variation, differing mainly in the suture curvature, chamber inflation, and umbilical side convexity, with two morphotypes: C. wuellerstorfi forma convexa, and C. wuellerstorfi forma plana. Cibicidoides lobatulus showed differences in test shape, primarily attributed to chamber deformation from substrate fixation adaptation. Cibicidoides ungerianus is reported for the first time in the Southeast Pacific, characterized by a keeled, primarily biconvex test, inflated chambers, coarse pores on both sides, transparent test, and glassy sutures. While Planulina ariminensis and P. ornata may resemble C. wuellerstorfi, they can be distinguished by their evolute and nearly parallel umbilical and spiral sides, a large planispiral test for the former and raised umbilical sutures for the latter. Planulina limbata stands out from the other species described herein due to its raised sutures, involute umbilical side, and elongated, narrow chambers on the umbilical side. Therefore, species of Cibicidoides and Planulina might exhibit similarities; however, significant morphological differences take place among them. The criteria developed here will serve as essential tools for future applications in the Southeast Pacific, particularly for taxonomy-based proxy calibrations and paleoceanographic reconstructions.