Molecular-based Methods Research Articles

Filarial parasite infection prevalence in field-caught mosquitoes from Connecticut, United States.

Filarial nematodes are parasitic roundworms transmitted by mosquitoes that can cause morbidity and mortality for their human and animal hosts. The filariae community, specifically infection prevalence of heartworm, Dirofilaria immitis (Filarioidea: Onchocercidae) (Leidy), and its primary mosquito vector species, has not been described in Connecticut since 1977. In light of the recent invasion and establishment of an important filariasis vector, Aedes albopictus (Diptera: Culicidae) (Skuse), we used molecular-based sequencing methods to identify filarial species infecting field-caught mosquitoes in Connecticut, United States. The filarial parasites identified include D. immitis, Aproctella sp., and Setaria sp. (Filarioidea: Setariidae). The total minimum infection rate for D. immitis for all mosquito species tested in 2020 was 0.97 [0.56 to 1.56] and in 2021 was 1.48 [0.93 to 2.24]. Aedes albopictus had the highest infection prevalence compared to other species during both years. We determined a low but persistent mosquito infection prevalence for D. immitis and suggest that Ae. albopictus is likely to be the primary vector in the region. Aproctella sp. and Setaria sp. had lower burdens compared to D. immitis. Persistent mosquito infection with filarial parasites, particularly in invasive species, poses a risk to veterinary and public health.

Improving Replication in Endometrial Omics: Understanding the Influence of the Menstrual Cycle.

The dynamic nature of human endometrial tissue presents unique challenges in analysis. Despite extensive research into endometrial disorders such as endometriosis and infertility, recent systematic reviews have highlighted concerning issues with the reproducibility of omics studies attempting to identify biomarkers. This review examines factors contributing to poor reproducibility in endometrial omics research. Hormonal fluctuations in the menstrual cycle lead to widespread molecular changes in the endometrium, most notably in gene expression profiles. In this review, we examine the variability in omics data due to the menstrual cycle and highlight the importance of accurate menstrual cycle dating for effective statistical modelling. The current standards of endometrial dating lack precision and we make the case for using molecular-based modelling methods to estimate menstrual cycle time for endometrium tissue samples. Additionally, we discuss statistical considerations such as confounding and interaction effects, as well as the importance of recording the detailed and accurate clinical information of patients. By addressing these methodological challenges, we aim to establish more robust and reproducible research practises, increasing the reliability of endometrial omics research and biomarker discovery.

Integrative taxonomy of the genus Pseudoacanthocephalus (Acanthocephala: Echinorhynchida) in China, with the description of two new species and the characterization of the mitochondrial genomes of Pseudoacanthocephalus sichuanensis sp. n. and Pseudoacanthocephalus nguyenthileae

BackgroundAcanthocephalans (thorny headed worms) of the genus Pseudoacanthocephalus mainly parasitize amphibians and reptiles across the globe. Some species of the genus Pseudoacanthocephalus also can accidentally infect human and cause human acanthocephaliasis. Current knowledge of the species composition of the genus Pseudoacanthocephalus from amphibians and reptiles in China is incomplete. An insufficiency of genetic data on species of the genus Pseudoacanthocephalus, including the complete mitochondrial genomes, has limited the use of molecular-based methods to better define the taxonomy and phylogeny of the genus Pseudoacanthocephalus. A more rigorous molecular phylogeny with broader representatives of the genus Pseudoacanthocephalus is required to further clarify the systematic status of the family Pseudoacanthocephalidae.MethodsMany specimens of the genus Pseudoacanthocephalus collected from toads and frogs in China were identified to species level using integrated morphological methods (light and scanning electron microscopy) and molecular approaches (sequencing different nuclear and mitochondrial genetic markers). The Assemble Species by Automatic Partitioning (ASAP) and Bayesian inference (BI) methods were applied for species delimitation. The complete mitochondrial genomes of two Pseudoacanthocephalus species were also sequenced and annotated to enrich the body of mitogenomic data on acanthocephalans. Additionally, phylogenetic analyses based on the amino acid sequences of 12 protein-coding genes (PCGs) of mitochondrial genomes of acanthocephalans using maximum likelihood (ML) and BI were performed to further investigate the phylogenetic position of the family Pseudoacanthocephalidae in the order Echinorhynchida.ResultsThree Pseudoacanthocephalus species, including P. sichuanensis sp. n., P. previatesticulus sp. n. and P. nguyenthileae were described. The results of ASAP and BI analyses based on the cytochrome c oxidase subunit 1 and subunit 2 (cox1, cox2) and 12S ribosomal RNA (12S) sequences supported the separation of P. sichuanensis and P. previatesticulus from the congeneric species. The results of BI inference using the internal transcribed spacer (ITS), cox1, cox2 and 12S sequence data indicated that P. sichuanensis and P. nguyenthileae have a closer relationship than P. previatesticulus and P. bufonis in Pseudoacanthocephalus. The complete mitogenomes of P. sichuanensis and P. nguyenthileae have 15,812 and 13,701 bp, respectively, with both including 36 genes and two non-coding regions. Phylogenetic results based on mitogenomic data demonstrated that the two families Pseudoacanthocephalidae and Arhythmacanthidae have a sister relationship in the order Echinorhynchida.ConclusionsTwo new species of the genus Pseudoacanthocephalus, namely P. sichuanensis sp. n. and P. previatesticulus sp. n., were identified based on integrated evidence. This is the first report of P. nguyenthileae in China. A revised key for the species of the genus Pseudoacanthocephalus was provided. Molecular analyses revealed that the mitochondrial cox1, cox2 and 12S genes as genetic markers seem to be more suitable for species delimitation of Pseudoacanthocephalus than the nuclear ITS region. BI results suggested a close affinity between P. sichuanensis and P. nguyenthileae. The mitochondrial genomic data of P. sichuanensis and P. nguyenthileae are provided for the first time. Mitogenomic phylogenetic results further confirmed the validity of the family Pseudoacanthocephalidae.Graphical

The Effect of MiR320a on Lung Cancer.

Lung cancer has a high mortality rate among cancers in both women and men. Currently, lung cáncer diagnosis is made with clinical examination, low-dose CT scan and molecular-based methods and its treatment options include chemotherapy, surgery, radiotherapy or immunotherapy. However, the life expectancy of lung cancer is not very high, and still it is usually diagnosed very lately, which leads to poorer prognosis. MicroRNAs [miRNAs] are small noncoding RNAs that regulate many diverse activities in the cell that can affect tumorigenesis by regulating many cell functions related to cancer, such as cell cycle, metastasis, angiogenesis, metabolism, and apoptosis. Also, it can have a potential diagnostic, therapeutic, and prognostic value for lung cancer. MiR320a is a promising microRNA that may help us in the diagnosis, treatment and prognosis of lung cancer, but some aspects of its clinical application are still vague, especially its effect on heavy smokers, delivery mechanism, toxicity and lack of reliable critical value. In this paper, we examined its comprehensive molecular interactions that lead to its tumor suppressor effect, and we reviewed its clinical application until now.

Systematic review and meta-analysis protocol: Diagnostic accuracy assessment for soil-transmitted helminth using microscopy and molecular-based methods

Soil-transmitted helminth (STH) is a part of NTDs that remains to be eliminated because of its significant global impact. Diagnostics are essential in aiding the elimination of STH infection. Therefore, it is imperative to evaluate the progress of diagnostic techniques, especially molecular-based techniques. Nevertheless, there is currently no comprehensive systematic review and meta-analysis (SRMA) evaluating the performance of molecular-based techniques for diagnosing STH. This protocol, registered on PROSPERO (CRD42024563432), intends to assess the diagnostic accuracy of molecular-based techniques compared to microscopic-based techniques for four common STH infections (A. lumbricoides, T. trichiura, hookworms, and S. stercoralis) across global studies. Two authors will independently conduct the screening, study selection, data extraction, and quality assessment of this SRMA. The obtained studies will be assessed using QUADAS-2 and will be analyzed for publication bias and diagnostic accuracy, including sensitivity, specificity, NPV, PPV, and DOR, and SROC. This developed protocol is expected to be versatile enough to be implemented in the range of studies assessing the diagnostic accuracy of the advance compared to the convention methods with adjustments. The findings of this SRMA are expected to provide a valuable contribution towards achieving the goal of eradicating STH infection by 2030.

Culicoides biting midges feeding behaviour as a key for understanding avian Haemoproteus transmission in Lithuania.

Investigations of host feeding behaviour in haematophagous insects are critical to assess transmission routes of vector-borne diseases. Understanding if a certain species has ornithophilic or mammalophilic feeding behaviour can facilitate future studies focused on pathogens transmission to and from certain host species. Culicoides Latreille (Diptera: Ceratopogonidae) are vectors of several pathogens, which include arboviruses, bacteria and parasites to a considerable diversity of vertebrate hosts. However, most of the studies focused on feeding habits target Culicoides species that could transmit the Bluetongue virus, consequently with a mammalophilic feeding behaviour, leaving aside the Culicoides species that are involved in the transmission of vector-borne parasites to birds, such as Haemoproteus Kruse (Haemosporida: Haemoproteidae). This study aimed to investigate the source of blood meals of wild-caught Culicoides using molecular-based methods and to correlate our findings with the reports of Haemoproteus parasites in Culicoides species. Engorged Culicoides females were collected using ultraviolet (UV)-light traps at seven different localities in Lithuania in 2021-2023. Biting midges were dissected, and the abdomens of engorged females were used for molecular investigation of the blood meal source. A polymerase chain reaction (PCR) protocol that amplifies a fragment of the Cytochrome B gene of vertebrates was used. Obtained sequences were compared to available information in GenBank database to confirm the source of the blood meal. In total, 258 engorged Culicoides females, representing nine different species, were analysed. The source of blood meal was identified in 29.1% of them with most of the insects having fed on birds (74.7%) and the remaining on mammals (25.3%). Culicoides segnis Campbell, Pelham-Clinton was the only species to feed exclusively on birds; Culicoides from the Obsoletus group, C. pallidicornis Kieffer and C. punctatus Latreille were found to feed exclusively on mammals; C. festivipennis Kieffer, C. kibunensis Tokunaga and C. pictipennis Staeger had an opportunistic feeding behaviour, with the first two preferably feeding on birds. Due to their feeding behaviour and the presence of Haemoproteus parasites reported in the literature, C. festivipennis, C. kibunensis, C. pictipennis, and C. segnis play an important role in the transmission of those avian vector-borne parasite in the wild. These Culicoides species were already confirmed as being able to support the development of several Haemoproteus species and lineages. Future studies focused on understanding the epidemiology of avian pathogens transmitted by Culicoides should target these species.

Molecular prevalence and distribution of tick-borne bacterial and protozoan pathogens of sheep and goats in Africa: A systematic review and meta-analysis

Tick-borne pathogens (TBPs) are a major impediment to the health, welfare, and production of small ruminants across the world, including Africa. Comprehensive information about the epidemiology of TBPs infecting sheep and goats across Africa is lacking. Therefore, this study was undertaken to determine the prevalence through a meta-analysis of selected TBPs in blood DNA from domestic sheep and goats in Africa obtained using molecular-based methods. The literature review was done according to Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) guidelines using five English electronic databases (PubMed, Science Direct, Springer Link, Web of Science, and African Journals Online (AJOL). The search was performed with no restriction in time through to 18th January 2023. Of the 63 full-text articles subjected to eligibility, only 30 articles met the eligibility criteria and were included in the review. The overall pooled prevalence of selected TBPs varied considerably between host species (sheep vs. goats), with Anaplasma ovis (44.50 vs. 48.40%), Ehrlichia ruminantium (5.50 vs. 2.00%), Coxiella burnetii (4.40 vs. 1.70%), Borrelia theileri in sheep (5.20%), Babesia ovis (1.70% vs. 1.90%), Theileria ovis (40.50% vs 10.00%), T. separata (1.00% vs 1.00%) and T. lestoquardi in sheep (8.40%). However, the prevalence of the selected TBPs was generally higher in sheep compared to goats. Several genetic loci were targeted in the characterization of tick-borne pathogens, such as 16S rDNA, groEL, and msp4 for Anaplasma ovis, pCS20 for Ehrlichia ruminantium, Insertion Sequence (IS1111) for Coxiella burnetii, flaB (flagellin) and 16S rRNA for Borrelia theileri, 5.8S rRNA and 18S rRNA for Babesia/Theileria, as well as the utilization of numerous PCR variants including conventional polymerase chain reaction (PCR), nested-PCR, qPCR, Loop-mediated isothermal amplification (LAMP), and reverse line blotting (RLB). In conclusion, A. ovis was the most widely distributed and prevalent TBP affecting small ruminants within the continent. Hence, this warrants adequate attention towards early diagnosis and treatment of infected animals as well as the control of the tick vectors involved in their transmission.

Prediction of Vacuum Ultraviolet/Ultraviolet Gas-Phase Absorption Spectra Using Molecular Feature Representations and Machine Learning.

Ultraviolet (UV) absorption spectroscopy is a widely used tool for quantitative and qualitative analyses of chemical compounds. In the gas phase, vacuum UV (VUV) and UV absorption spectra are specific and diagnostic for many small molecules. An accurate prediction of VUV/UV absorption spectra can aid the characterization of new or unknown molecules in areas such as fuels, forensics, and pharmaceutical research. An alternative to quantum chemical spectral prediction is the use of artificial intelligence. Here, different molecular feature representation techniques were used and developed to encode chemical structures for testing three machine learning models to predict gas-phase VUV/UV absorption spectra. Structure data files (.sdf) and VUV/UV absorption spectra for 1397 volatile and semivolatile chemical compounds were used to train and test the models. New molecular features (termed ABOCH) were introduced to better capture pi-bonding, aromaticity, and halogenation. The incorporation of these new features benefited spectral prediction and demonstrated superior performance compared to computationally intensive molecular-based deep learning methods. Of the machine learning methods, the use of a Random Forest regressor returned the best accuracy score with the shortest training time. The developed machine learning prediction model also outperformed spectral predictions based on the time-dependent density functional theory.

Evaluation of molecular-based methods for the detection and quantification of Cryptosporidium spp. in wastewater

Evaluation of molecular-based methods for the detection and quantification of Cryptosporidium spp. in wastewater

Which Polymorphism Is The Distinguishing Factor For Fitness Athletes: The Ace Indel or Actn3 rs1815739?

Purpose: The purpose of this research is to investigate the occurrence of ACE I/D and ACTN3 rs1815739 variations in sub-elite fitness athletes and to determine which gene is distinctive by comparing genotype and allele frequencies with sedentary individuals. Material and Methods: 41 sub-elite fitness athletes and 41 sedentary volunteers participated in the study. Genetic analyzes of the athletes were determined using molecular-based methods such as DNA isolation using a commercial kit, Real-time polymerase chain reaction (Real-Time PCR), and Polymerase chain reaction (PCR) after blood collection in EDTA tubes. The Chi-Square test was used to compare the genotype distribution and I, D, C, and T allele frequencies of ACE I/D and ACTN3 rs1815739 polymorphisms for statistical significance. Results: Analysis of data from our study revealed no statistically significant differences in ACE I/D polymorphisms between groups (p=0.4438), but statistically significant differences in ACTN3 rs1815739 polymorphisms significant difference (p=0.0313). Conclusion: In conclusion, in our study, it can be said that the ACTN3 gene is a distinguishing factor in fitness.

Culture-negative periprosthetic joint infections: Do we have an issue?

Culture-negative periprosthetic joint infections: Do we have an issue?

Artificial Intelligence Plant Doctor: Plant Disease Diagnosis Using GPT4-vision

Integrated pest management is essential for controlling plant diseases that reduce crop yields. Rapid diagnosis is crucial for effective management in the event of an outbreak to identify the cause and minimize damage. Diagnosis methods range from indirect visual observation, which can be subjective and inaccurate, to machine learning and deep learning predictions that may suffer from biased data. Direct molecular-based methods, while accurate, are complex and time-consuming. However, the development of large multimodal models, like GPT-4, combines image recognition with natural language processing for more accurate diagnostic information. This study introduces GPT-4-based system for diagnosing plant diseases utilizing a detailed knowledge base with 1,420 host plants, 2,462 pathogens, and 37,467 pesticide instances from the official plant disease and pesticide registries of Korea. The AI plant doctor offers interactive advice on diagnosis, control methods, and pesticide use for diseases in Korea and is accessible at https://pdoc.scnu.ac.kr/.

High Abundance of Haemoproteus Parasites in Culicoides (Diptera, Ceratopogonidae), with a Confirmation of Culicoides reconditus as a New Vector of These Avian Blood Parasites.

Haemoproteus parasites are the most diverse among Haemosporida. However, their natural vectors (Culicoides) are still poorly investigated and were identified for only a few parasite species and lineages. The application of an integrative approach (insect dissection, microscopic analysis, and molecular-based methods) is necessary in these studies, which have been carried out by a few research groups, mainly in Europe. The aim of this study was (i) to determine the Culicoides species that are naturally infected by Haemoproteus parasites, and which can support its complete sporogonic development, and (ii) to investigate the prevalence of Culicoides species and Haemoproteus parasite lineages in different study sites. In total, 1953 parous Culicoides females, from 11 species, were collected in four different localities in Lithuania and were dissected and analyzed using an integrative approach. The most abundant was C. pictipennis (30.3%). Parasite DNA was found in 7.9% of all investigated Culicoides, of which ~30% had sporozoites in their salivary glands, confirming their vector competence for these parasites. The Botanical Garden presented the highest number of Culicoides parous females, Culicoides species, and parasite lineages, as well as the highest positivity for sporozoites. Culicoides reconditus was confirmed as a natural vector of Haemoproteus parasites, sporozoites of six Haemoproteus lineages were reported for the first time, and 12 new interactions between Haemoproteus parasite lineages and Culicoides species were identified. Haemoproteus parasites seem to be transmitted by a high number of Culicoides species, with C. kibunensis, C. pictipennis, and C. segnis being the most important vectors.

Meta-analysis of Johne's disease in the Iranian animal population (1999-2020).

Johne's disease (JD) affects domestic and wild animals across the globe. Paratuberculosis exerts huge economic impacts on the animal industry. Despite significant economic losses, little knowledge is available on the epidemiological status of Paratuberculosis in the animal population of Iran. The present study aimed to evaluate the prevalence rate of this disease in the Iranian animal population with confidence interval (CI) and p-value. The search was conducted on and screened the electronic international and national databases. Thereafter, sufficient and relevant data were extracted. Data were analyzed in STATA software (version 14). Prevalence disease rates were determined using random effect models. A total of 52 articles were included in the systematic review. According to the results, the overall disease incidence rate in Iran was 20.39%. The prevalence rate of JD was 22.33% (95% CI, 18.87-25.78) in the cattle population and 25.61% (95% CI, 21.43-29.78) in sheep. This study pinpointed that cattle and sheep were the most commonly infected hosts. The highest prevalence rate of disease was 35.88% in Tehran (95% CI, 16.77-54.99), followed by 32.86% (95% CI, 25.07-40.65), and 20.10% (95% CI, 14.63-25.58) in Khorasan Razavi and Kerman, respectively. The lowest prevalence rate of JD was 2.27% in Ilam (95% CI, 0.84-3.70). Based on this result, molecular-based methods were properly compared to other diagnostic methods. This study reported Mycobacterium avium subsp. paratuberculosis (MAP) prevalence in dairy herds in the provinces of Iran. The infection transmission from animal sources to humans and the potential role of MAP in human disease highlight a critical need for further study on this issue.

Diversity of antibiotic resistance gene variants at subsequent stages of the wastewater treatment process revealed by a metagenomic analysis of PCR amplicons.

Wastewater treatment plants have been recognised as point sources of various antibiotic-resistant bacteria (ARB) and antibiotic resistance genes (ARG) which are considered recently emerging biological contaminants. So far, culture-based and molecular-based methods have been successfully applied to monitor antimicrobial resistance (AMR) in WWTPs. However, the methods applied do not permit the comprehensive identification of the true diversity of ARGs. In this study we applied next-generation sequencing for a metagenomic analysis of PCR amplicons of ARGs from the subsequent stages of the analysed WWTP. The presence of 14 genes conferring resistance to different antibiotic families was screened by PCR. In the next step, three genes were selected for detailed analysis of changes of the profile of ARG variants along the process. A relative abundance of 79 variants was analysed. The highest diversity was revealed in the ermF gene, with 52 variants. The relative abundance of some variants changed along the purification process, and some ARG variants might be present in novel hosts for which they were currently unassigned. Additionally, we identified a pool of novel ARG variants present in the studied WWTP. Overall, the results obtained indicated that the applied method is sufficient for analysing ARG variant diversity.

Molecular characterization of antimicrobial resistance genes of Staphylococcus aureus isolated from mastitic camel milk in Egypt.

Staphylococcus aureus is one of the most common causes of mastitis worldwide. This study aimed to determine the prevalence and antimicrobial resistance (AMR) patterns of S. aureus in mastitic milk samples collected from camel farms in Matrouh Governorate, Egypt. A total of 200 mastitic camel milk samples were evaluated for S. aureus using both conventional culture-based and molecular-based methods. Antibiotic susceptibility testing of S. aureus isolates was conducted using disc diffusion and agar dilution methods, with antibiotic resistance genes identified through polymerase chain reaction with specific primers. Out of samples tested, 60 (30.00%) were positive for S. aureus. The isolates displayed the highest of resistance against piperacillin-tazobactam (55.00%) followed by trimethoprim- sulfamethoxazole (45.00%) and amoxicillin (40.00%). Half of the isolates were multidrug-resistant (MDR). The AMR genes included methicillin-resistant gene (mecA), β-lactamase gene (blaZ), tetracycline resistance gene (tetK), erythromycin resistance gene (ermB) and vancomycin resistant gene (vanA) were detected in 100%, 100%, 95.00%, 90.00% and 20.00% of the isolates, respectively. In conclusion, the presence of MDRS aureus as a cause of clinical camel mastitis is a significant veterinary and public health concern. These findings highlight the importance of proper antibiotic use in Egyptian camel farms and the need for molecular techniques to fully understand the genetic profile of antimicrobial-resistant S. aureus isolates.

Molecular Techniques for Root-Knot Nematode Identification.

Among plant-parasitic nematodes, root-knot nematodes (RKN), Meloidogyne spp., are the most important parasite infecting economically important crops globally and causing severe losses in crop production. The use of efficient nematode control methods against these parasites depends upon their correct detection in roots and soil samples. Currently, the use of integrated identification methods, including biochemical, molecular, and morphological-based characters, is preferred. But the techniques using morphology and phylogenetic analysis are time-consuming and not suitable for routine analysis. They have only been used for studies of cryptic species, which were identified using integrative taxonomy. Here we describe the enzymatic and molecular-based methods that have successfully been used in Brazil for more than 25years in the Nematology Lab at Embrapa Genetic Resources and Biotechnology for routine analysis. This technique is a combination of isozyme esterase profiling and molecular markers, with the aim of having a rapid and correct diagnosis of Meloidogyne spp. populations from field and greenhouse.

Molecular Approaches to Measure Bacterial Extracellular Vesicles in Tissue Samples.

Extracellular vesicles (EVs) are naturally occurring lipid-bound nanoparticles produced by all cell types. Growing work demonstrates the ability of EVs to facilitate long-distance and cross-kingdom communication. Their innate barrier crossing and cell targeting properties make them a uniquely useful starting ground for novel drug delivery platforms. To better understand the endogenous activity and therapeutic potential of EVs, recent work has measured particle circulation and distribution in vivo using several approaches. Here, we describe molecular-based methods for quantifying bacterial EV distribution in collected tissue samples for biodistribution studies. These methods are important for understanding cell-cell communication facilitated by bacterial EVs and for identifying opportunities for using bacterial EVs as a therapeutic platform.

Salmonella enterica Serovar Gallinarum Biovars Pullorum and Gallinarum in Poultry: Review of Pathogenesis, Antibiotic Resistance, Diagnosis and Control in the Genomic Era.

Salmonella enterica subsp. enterica serovar Gallinarum (SG) has two distinct biovars, Pullorum and Gallinarum. They are bacterial pathogens that exhibit host specificity for poultry and aquatic birds, causing severe systemic diseases known as fowl typhoid (FT) and Pullorum disease (PD), respectively. The virulence mechanisms of biovars Gallinarum and Pullorum are multifactorial, involving a variety of genes and pathways that contribute to their pathogenicity. In addition, these serovars have developed resistance to various antimicrobial agents, leading to the emergence of multidrug-resistant strains. Due to their economic and public health significance, rapid and accurate diagnosis is crucial for effective control and prevention of these diseases. Conventional methods, such as bacterial culture and serological tests, have been used for screening and diagnosis. However, molecular-based methods are becoming increasingly important due to their rapidity, high sensitivity, and specificity, opening new horizons for the development of innovative approaches to control FT and PD. The aim of this review is to highlight the current state of knowledge on biovars Gallinarum and Pullorum, emphasizing the importance of continued research into their pathogenesis, drug resistance and diagnosis to better understand and control these pathogens in poultry farms.

Antimicrobial Resistance Profile of Planctomycetota Isolated from Oyster Shell Biofilm: Ecological Relevance within the One Health Concept

Background: Planctomycetota isolation in pure culture is still challenging with most of the reported data coming from molecular-based methods. Here, we intended to isolate Planctomycetota from the filter-feeder Pacific oyster Magallana gigas, extending the search to a not yet explored natural reservoir and to characterize their antimicrobial resistance phenotype. Methods: Oyster samples from different supermarkets and from a farm producer were subject to isolation in selective medium. Inoculation was performed from the shell biofilm and after an enrichment of the edible content. Results: Planctomycetota isolates (n = 65) were only obtained from the shell biofilm with four different species identified: Rhodopirellula baltica (n = 62), Rhodopirellula rubra (n = 1), Rhodopirellula heiligendammensis (n = 1) and Gimesia chilikensis (n = 1). This study reports the first association of Planctomycetota members with oysters and the first description of R. heiligendammensis in Portugal. Moreover, R. rubra, originally identified in Portugal, was isolated from oysters of French origin. Antibiotic susceptibility testing, conducted in strains belonging to two species never assayed before revealed multidrug resistance phenotypes with bacteria showing resistance to several classes of clinically relevant antibiotics (e.g., β-lactams and aminoglycosides). Conclusion: The ecological role and impact of Planctomycetota on oyster holobiont and, ultimately, in public health, under the One Health concept, is discussed.