Targeted volume correlative light and electron microscopy of an environmental marine microorganism.

Photosynthetic microalgae are responsible for an important fraction of CO2 fixation and O2 production on Earth. Three-dimensional (3D) ultrastructural characterization of these organisms in their natural environment can contribute to a deeper understanding of their cell biology. However, the low throughput of volume electron microscopy (vEM) methods along with the complexity and heterogeneity of environmental samples pose great technical challenges. In the present study, we used a workflow based on a specific electron microscopy sample preparation method compatible with both light and vEM imaging in order to target one cell among a complex natural community. This method revealed the 3D subcellular landscape of a photosynthetic dinoflagellate, which we identified as Ensiculifera tyrrhenica, with quantitative characterization of multiple organelles. We show that this cell contains a single convoluted chloroplast and show the arrangement of the flagellar apparatus with its associated photosensitive elements. Moreover, we observed partial chromatin unfolding, potentially associated with transcription activity in these organisms, in which chromosomes are permanently condensed. Together with providing insights in dinoflagellate biology, this proof-of-principle study illustrates an efficient tool for the targeted ultrastructural analysis of environmental microorganisms in heterogeneous mixes.

Eco-friendly solvents in liquid-liquid microextraction techniques for biological and environmental analysis: a critical review.

In recent years, green solvents have emerged as promising alternatives in the field of analytical chemistry, replacing conventional organic solvents known for their toxicity, volatility, and flammability. The combination of these solvents with liquid-liquid microextraction techniques has facilitated the development of simpler, faster, more economical, and environment-friendly methodologies for the analysis of samples of varying complexity. This review discusses the fundamental physicochemical properties and advantages of using deep eutectic solvents, ionic liquids, switchable-hydrophilicity solvents, supramolecular solvents, and surfactants as extractants. Furthermore, analytical methods based on liquid-liquid microextraction techniques developed in the last 5 years for the determination of organic compounds and metals in biological and environmental samples are presented and discussed, highlighting their applications and benefits to improve analytical performance and sustainability.

The newest Oxford Nanopore R10.4.1 full-length 16S rRNA sequencing enables the accurate resolution of species-level microbial community profiling.

The long-read amplicon provides a species-level solution for the community. With the improvement of nanopore flowcells, the accuracy of Oxford Nanopore Technologies (ONT) R10.4.1 has been substantially enhanced, with an average of approximately 99%. To evaluate its effectiveness on amplicons, three types of microbiomes were analyzed by 16S ribosomal RNA (hereinafter referred to as "16S") amplicon sequencing using Novaseq, Pacbio sequel II, and Nanopore PromethION platforms (R9.4.1 and R10.4.1) in the current study. We showed the error rate, recall, precision, and bias index in the mock sample. The error rate of ONT R10.4.1 was greatly reduced, with a better recall in the case of the synthetic community. Meanwhile, in different types of environmental samples, ONT R10.4.1 analysis resulted in a composition similar to Pacbio data. We found that classification tools and databases influence ONT data. Based on these results, we conclude that the ONT R10.4.1 16S amplicon can also be used for application in environmental samples. IMPORTANCE The long-read amplicon supplies the community with a species-level solution. Due to the high error rate of nanopore sequencing early on, it has not been frequently used in 16S studies. Oxford Nanopore Technologies (ONT) introduced the R10.4.1 flowcell with Q20+ reagent to achieve more than 99% accuracy as sequencing technology advanced. However, there has been no published study on the performance of commercial PromethION sequencers with R10.4.1 flowcells on 16S sequencing or on the impact of accuracy improvement on taxonomy (R9.4.1 to R10.4.1) using 16S ONT data. In this study, three types of microbiomes were investigated by 16S ribosomal RNA (rRNA) amplicon sequencing using Novaseq, Pacbio sequel II, and Nanopore PromethION platforms (R9.4.1 and R10.4.1). In the mock sample, we displayed the error rate, recall, precision, and bias index. We observed that the error rate in ONT R10.4.1 is significantly lower, especially when deletions are involved. First and foremost, R10.4.1 and Pacific Bioscience platforms reveal a similar microbiome in environmental samples. This study shows that the R10.4.1 full-length 16S rRNA sequences allow for species identification of environmental microbiota.

Development of Simple Fluorescence Analytical Strategy for the Detection of Triazophos Using Greenish-Yellow Emissive Carbon Dots Derived from Curcuma longa.

This study describes a new method for synthesizing water-soluble carbon dots (CDs) using "Curcuma longa" (green source) named CL-CDs via a single-step hydrothermal process. The as-synthesized CL-CDs exhibited greenish-yellow fluorescence at 548 nm upon excitation at 440 nm. It shows good water stability and exhibits a quantum yield of 19.4%. The developed probe is utilized for sensing triazophos (TZP) pesticide via a dynamic quenching mechanism, exhibiting favorable linearity ranging from 0.5-500 µM with a limit of detection of 0.0042 µM. The as-prepared CL-CDs probe was sensitive and selective towards TZP. Lastly, the successful application of the CL-CDs-based fluorescent probe in water and rice samples highlights its potential as a reliable and efficient method for the detection of TZP in various real sample matrices. Eventually, bioimaging and biocompatibility aspects of CL-CDs have been assessed on Saccharomyces cerevisiae (yeast) cell and lung cancer (A549) cell lines, respectively.

The snow alga Chloromonas kaweckae sp. nov. (Volvocales, Chlorophyta) causes green surface blooms in the high tatras (Slovakia) and tolerates high irradiance.

Seasonally slowly melting mountain snowfields are populated by extremophilic microalgae. In alpine habitats, high-light sensitive, green phytoflagellates are usually observed in subsurface layers deeper in the snowpack under dim conditions, while robust orange to reddish cyst stages can be seen exposed on the surface. In this study, uncommon surface green snow was investigated in the High Tatra Mountains (Slovakia). The monospecific community found in the green surface bloom consisted of vegetative Chloromonas cells (Volvocales, Chlorophyta). Molecular data demonstrated that the field sample and the strain isolated and established from the bloom were conspecific, and they represent a new species, Chloromonas kaweckae sp. nov., which is described based on the morphology of the vegetative cells and asexual reproduction and on molecular analyses of the strain. Cells of C. kaweckae accumulated approximately 50% polyunsaturated fatty acids, which is advantageous at low temperatures. In addition, this new species performed active photosynthesis at temperatures close to the freezing point showed a light compensation point of 126 ± 22 µmol photons · m-2  · s-1 and some signs of photoinhibition at irradiances greater than 600 µmol photons · m-2  · s-1 . These data indicate that the photosynthetic apparatus of C. kaweckae could be regarded as adapted to relatively high light intensities, otherwise unusual for most flagellate stages of snow algae.

Multiple heteroatom dopant carbon dots as a novel photoluminescent probe for the sensitive detection of Cu2+ and Fe3+ ions in living cells and environmental sample analysis.

Heavy metal ion pollution harms human health and the environment and continues to worsen. Here, we report the synthesis of boron (B), phosphorous (P), nitrogen (N), and sulfur (S) co-doped carbon dots (BP/NS-CDs) by a one-step facile hydrothermal process. The optimum synthetic parameters are of 180 °C temperature, 12 h reaction time and 15% of PBA mass. The as-synthesized BP/NS-CDs exhibits excellent water solubility, strong green photoluminescence (PL) at 510 nm, and a high quantum yield of 22.4%. Moreover, BP/NS-CDs presented high monodispersity (7.2 ± 0.45 nm), excitation-dependent emission, PL stability over large pH, and high ionic strength. FTIR, XRD, and XPS are used to confirm the successful B and P doping of BP/NS-CDs. BP/NS-CD photoluminescent probes are selectively quenched by Cu2+ and Fe3+ ions but showed no response to the presence of other metal cations. The PL emission of BP/NS-CDs exhibited a good linear correlation with Cu2+ and Fe3+ concentrations with detection limits of 0.18 µM and 0.27 µM for Cu2+ and Fe3+, respectively. Furthermore, the HCT116 survival cells kept at 99.4 ± 1.3% and cell imaging capability, when the BP/NS-CDs concentration is up to 300 µg/mL by MTT assay. The proposed sensor is potential applications for the detection of Cu2+ and Fe3+ ions in environmental water samples.

Evaluation of two methods for detection of viable Bacillus anthracis simulant spores in maritime environmental samples.

Analytical methods exist to detect biothreat agents in environmental samples during a response to biological contamination incidents. However, the coastal zone facilities and assets of the US Coast Guard (USCG), including response boats in diverse geographical areas and maritime environmental conditions, can pose complex and unique challenges for adapting existing analytical detection methods. The traditional culture (TC) and the rapid viability polymerase chain reaction (RV-PCR) methods were evaluated for their compatibility for maritime environmental surface and grab sample analysis to detect spores of Bacillus thuringiensis subspecies kurstaki (Btk), a surrogate for Bacillus anthracis. The representative samples collected from a USCG installation included surfaces, such as aluminum on boats, nonskid tread on decks of watercraft, computer touchscreens, and concrete piers, and grab samples of boat washdown water, soil, vegetation, and gravel from surrounding areas. Replicate samples were spiked with Btk spores at two to three tenfold increasing levels and analyzed. Out of a total of 150 samples collected and analyzed, the TC method gave 10 false-positive and 19 false-negative results, while the RV-PCR method-based analysis resulted in 0 false-positive and 26 false-negative results. An abundance of microbial background and particulates in some samples interfered with true results, while both methods gave similar results for samples with low microbial background and particulates. Improved and high-throughput sample processing methods are needed for analysis of complex environmental samples.

Analysis of micro- and nanoplastics in wastewater treatment plants: key steps and environmental risk considerations.

The analysis of micro- and nanoplastics (MNPs) in the environment is a critical objective due to their ubiquitous presence in natural habitats, as well as their occurrence in various food, beverage, and organism matrices. MNPs pose significant concerns due to their direct toxicological effects and their potential to serve as carriers for hazardous organic/inorganic contaminants and pathogens, thereby posing risks to both human health and ecosystem integrity. Understanding the fate of MNPs within wastewater treatment plants (WWTPs) holds paramount importance, as these facilities can be significant sources of MNP emissions. Additionally, during wastewater purification processes, MNPs can accumulate contaminants and pathogens, potentially transferring them into receiving water bodies. Hence, establishing a robust analytical framework encompassing sampling, extraction, and instrumental analysis is indispensable for monitoring MNP pollution and assessing associated risks. This comprehensive review critically evaluates the strengths and limitations of commonly employed methods for studying MNPs in wastewater, sludge, and analogous environmental samples. Furthermore, this paper proposes potential solutions to address identified methodological shortcomings. Lastly, a dedicated section investigates the association of plastic particles with chemicals and pathogens, alongside the analytical techniques employed to study such interactions. The insights generated from this work can be valuable reference material for both the scientific research community and environmental monitoring and management authorities.

A dual-channel probe based on copper ion-mediated metal organic framework composite for colorimetric and ratiometric fluorescence monitoring of glyphosate degradation in soil and water.

A dual-channel probe was developed, based on a novel composite metal organic frameworks (ZnMOF-74@Al-MOF) for glyphosate determination through ratio fluorescence and colorimetric methods. The prepared probe can not only recognize and combine glyphosate by introducing copper ion into the MOF, but also possess peroxidase-like catalytic activity. The recognition of target glyphosate brought about changes relative to its concentration on fluorescence intensity and ultraviolet absorption. And, the high specific surface area and porosity of porphyrin MOF provides the developed probe with more response opportunities to afford a better detection performance for glyphosate. Under optimum conditions, the copper ion-mediated method exhibited good detection performance for glyphosate with low detection limits (0.070 and 0.092 µg mL-1 for fluorescence and colorimetric techniques, respectively). Furthermore, the possible mechanisms of the fluorescence quenching and the peroxidase-like catalytic of the probe were also explored. This dual-channel method was applied to monitor glyphosate degradation in environmental samples and satisfactory results were obtained.

Determination of 241Am in Environmental Samples: A Review.

The determination of 241Am in the environment is of importance in monitoring its release and assessing its environmental impact and radiological risk. This paper aims to give an overview about the recent developments and the state-of-art analytical methods for 241Am determination in environmental samples. Thorough discussions are given in this paper covering a wide range of aspects, including sample pre-treatment and pre-concentration methods, chemical separation techniques, source preparation, radiometric and mass spectrometric measurement techniques, speciation analyses, and tracer applications. The paper focuses on some hyphenated separation methods based on different chromatographic resins, which have been developed to achieve high analytical efficiency and sample throughput for the determination of 241Am. The performances of different radiometric and mass spectrometric measurement techniques for 241Am are evaluated and compared. Tracer applications of 241Am in the environment, including speciation analyses of 241Am, and applications in nuclear forensics are also discussed.

Analytical Methods for the Determination of 90Sr and 239,240Pu in Environmental Samples.

Artificial long-lived radionuclides such as 90Sr and 239,240Pu have been long released into the environment by human nuclear activities, which have a profound impact on the ecological environment. It is of great significance to monitor the concentration of these radionuclides for environmental safety. This paper summarizes and critically discusses the separation and measurement methods for ultra-trace determination of 90Sr, 239Pu, and 240Pu in the environment. After selecting the measurement method, it is necessary to consider the decontamination of the interference from matrix elements and the key elements, and this involves the choice of the separation method. Measurement methods include both radiometric methods and non-radiometric methods. Radiometric methods, including alpha spectroscopy, liquid scintillation spectrometry, etc., are commonly used methods for measuring 239+240Pu and 90Sr. Mass spectrometry, as the representative of non-radiometric measurement methods, has been regarded as the most promising analytical method due to its high absolute sensitivity, low detection limit, and relatively short sample-analysis time. Through the comparison of various measurement methods, the future development trend of radionuclide measurement is prospected in this review. The fully automatic and rapid analysis method is a highlight. The new mass spectrometer with ultra-high sensitivity shows strong analytical capabilities for extremely low concentrations of 90Sr, 239Pu, and 240Pu, and it is expected to develop determination methods with higher sensitivity and lower detection limit.

Strontium tungstate-modified disposable strip for electrochemical detection of sulfadiazine in environmental samples.

Rapid-monitoring of drugs has attracted tremendous consideration owing to robust global demand for cost-effective and high effectiveness. Binary metal oxides with various morphology have been reported as electrodes for electrochemical sensor to fulfilling the clinical and enviromental requirements. In this study, strontium tungstate (SrWO4) nanoflakes have been successfully prepared via the facile sonochemical method for the first time. The characteristics of as-prepared SrWO4 are systematically measured by various analytical and spectroscopic methods. The SrWO4 nanoflakes are utilized to modify the electrochemical electrode for the sulfadiazine (SDZ) determination. The SrWO4 modified electrode possesses excellent electrocatalytic activity and high recognition capability for the electrochemical detection of SDZ. Impressively, the as-fabricated SrWO4 modified electrode attainted lowest oxidation peak at +0.93 V (vs Ag/AgCl2) with the limit of detection of 0.009 µM, the sensitivity of 0.123 µA µM-1 cm2 and linear detection range of 0.05-235 µM. The enhanced performance of proposed SrWO4-based sensors could be attributed to the catalytic effect, large surface area, good electrical conductivity and physicochemical nature. Notably, the electrocatalytic performances of the SDZ sensors are good as compared to the previous literature, indicating the significance of the newly designed SrWO4 modified electrode. The real-sample diagnosis by the SDZ detection in environmental sample demonstrates the proposed SrWO4-based sensors with good recovery range.

Determination of sulfa antibiotic residues in river and particulate matter by field-amplified sample injection-capillary zone electrophoresis.

In the present research, field-amplified sample injection-CZE (FASI-CZE) coupled with a diode array detector was established to determine trace level sulfa antibiotic. Sulfathiazole, sulfadiazine, sulfamethazine, sulfadimethoxine, sulfamethoxazole, and sulfisoxazole were selected as analytes for the experiments. The background electrolyte solution consisted of 70.0 mmol/L borax and 60.0 mmol/L boric acid (including 10% methanol, pH 9.1). The plug was 2.5 mmol/L borax, which was injected into the capillary at a pressure of 0.5 psi for 5 s. Then the sample was injected into the capillary at an injection voltage of -10 kV for 20 s. The electrophoretic separation was carried out under a voltage of +19 kV. The capillary temperature was maintained at 20˚C throughout the analysis, and six sulfonamides were completely separated within 35 min. Compared with pressure injection-CZE, the sensitivity of FASI-CZE was increased by 6.25-10.0 times, and the LODs were reduced from 0.2-0.5 to 0.02-0.05 µg/mL. The method was applied to the determination of sulfonamides in river water and particulate matter samples. The recoveries were 78.59-106.59%. The intraday and interday precisions were 2.89-7.35% and 2.77-7.09%, respectively. This provides a simpler and faster method for the analysis of sulfa antibiotic residues in environmental samples.

Development of an advanced electrochemical biosensing platform for E. coli using hybrid metal-organic framework/polyaniline composite.

Because of numerous merits (e.g., the possibility of their synthesis in 1-D, 2-D, and 3-D forms, large surface-to-volume ratio, and flexible framework functionality), metal-organic frameworks (MOFs) are envisaged as excellent media for the development of biosensors for diverse analytes present in environmental media. The present research work, for the first time, reports the development of a Cu-MOF based electrochemical biosensor for highly sensitive detection of E. coli bacteria. In order to realize an MOF-based electrochemically active platform, Cu3(BTC)2 (BTC = 1,3,5-benzenetricarboxylic acid) was mixed with polyaniline (PANI). The spectroscopic/morphological characterizations of the resulting composite were established with the aid of FT-IR, UV-visible spectroscopy, X-ray diffraction, electron microscopy, and surface area analysis. The thin films of Cu3(BTC)2-PANI, on an indium-tin oxide (ITO) substrate, were bio-interfaced with anti-E. coli antibodies for use as a novel biosensing electrode. Based on the electrochemical impedance spectroscopy (EIS) technique of signal measurement, the above sensor exhibited high sensitivity to detect very low concentrations of E. coli (2cfu/mL) in a short response time (~2 min) and was also selective in the presence of other non-specific bacteria. As a novel highlight of the research, this new MOF/PANI based detection platform for E. coli has shown improved performance than many of the previously reported electrochemical biosensors.

Environmental sample characteristics and herd size associated with decreased herd-level prevalence of Mycobacterium avium ssp. paratuberculosis.

Environmental sampling is an effective method for estimating regional dairy herd-level prevalence of infection with Mycobacterium avium ssp. paratuberculosis (MAP). However, factors affecting prevalence estimates based on environmental samples are not known. The objective was to determine whether odds of environmental samples collected on farm changed culture status over 2 sampling times and if changes were specific for location and type of housing (freestall, tiestall, or loose housing), the sample collected (i.e., manure of lactating, dry, or sick cows; namely, cow group), and effects of herd size. In 2012-2013 [sampling 1 (S1)] and 2015-2017 [sampling 2 (S2)], 6 environmental samples were collected and cultured for MAP from all 167 (99%) and 160 (95%) farms, respectively, in the province of Saskatchewan, Canada. Only the 148 dairy farms sampled at both sampling periods were included in the analysis. A mixed effects logistic regression was used to determine whether differences between sampling periods were associated with herd size and sample characteristics (cow group contributing to environmental sample, type of housing, and location). In S1 and S2, 55 and 34%, respectively, of farms had at least 1 MAP-positive environmental sample. Correcting for sensitivity of environmental sampling, estimated true prevalence in S1 and S2 was 79 and 48%, respectively. Herds with >200 cows were more often MAP-positive than herds with <51 cows in both S1 and S2. The percentage of positive samples was lower in S2 compared with S1 for all sampled areas, cow groups contributing to samples, types of housing where samples were collected, and herd size categories. However, samples collected from dry cow areas had the largest decrease in MAP-positive samples in S2 compared with all other cow group samples. Herds that were MAP-negative in S1 with a herd size 51 to 100 or 101 to 150 were more likely to stay MAP-negative, whereas MAP-positive herds with >200 cows more frequently stayed MAP-positive. No difference was observed in the odds of a sample being MAP-positive among housing types or location of sample collection in both sample periods. Of all farms sampled, 104 (70%) did not change status from S1 to S2. In conclusion, when herd-level MAP prevalence decreased over the 3-yr interval, the change in prevalence differed among herd size categories and was larger in samples from dry cow areas. It was, however, not specific to other characteristics of environmental samples collected.

Use of Highly Specific Molecular Markers Reveals Positive Correlation between Abundances of Mesodinium cf. major and Its Preferred Prey, Teleaulax amphioxeia, During Red Water Blooms in the Columbia River Estuary.

In a previous study, Teleaulax amphioxeia-the preferred prey of Mesodinium in the Columbia River estuary-were undetectable within intense annual blooms, suggesting blooms are prey-limited or prey are acquired outside of bloom patches. We used a novel molecular approach specifically targeting the prey (i.e., Unique Sequence Element [USE] within the ribosomal RNA 28S D2 regions of T. amphioxeia nucleus and nucleomorph) in estuarine water samples acquired autonomously with an Environmental Sample Processor integrated within a monitoring network (ESP-SATURN). This new approach allowed for both more specific detection of the prey and better constraint of sample variability. A positive correlation was observed between abundances of M. cf. major and T. amphioxeia during bloom periods. The correlation was stronger at depth (> 8.2 m) and weak or nonexistent in the surface, suggesting that predator-prey dynamics become uncoupled when stratification is strong. We confirmed exclusive selectivity for T. amphioxeia by M. cf. major and observed the incorporation of the prey nucleus into a 4-nuclei complex, where it remained functionally active. The specific biomarker for T. amphioxeia was also recovered in M. cf. major samples from a Namibian coastal bloom, suggesting that a specific predator-prey relationship might be widespread between M. cf. major and T. amphioxeia.

Simultaneous monitoring of faecal indicators and harmful algae using an in-situ autonomous sensor.

UNLABELLED: Faecal indicator bacteria (FIB) and harmful algal blooms (HABs) threaten the health and the economy of coastal communities worldwide. Emerging automated sampling technologies combined with molecular analytical techniques could enable rapid detection of micro-organisms in-situ, thereby improving resource management and public health decision-making. We evaluated this concept using a robotic device, the Environmental Sample Processor (ESP). The ESP automates in-situ sample collection, nucleic acid extraction and molecular analyses. Here, the ESP measured and reported concentrations of FIB (Enterococcus spp.), a microbial source-tracking marker (human-specific Bacteriodales) and a HAB species (Psuedo-nitzschia spp.) over a 45-day deployment on the Santa Cruz Municipal Wharf (Santa Cruz, CA, USA). Both FIB and HABs were enumerated from single in-situ collected water samples. The in-situ qPCR efficiencies ranged from 86% to 105%, while the limit of quantifications during the deployment was 10 copies reaction(-1) . No differences were observed in the concentrations of enterococci, the human-specific marker in Bacteroidales spp., and P. australis between in-situ collected sample and traditional hand sampling methods (P > 0·05). Analytical results were Internet-accessible within hours of sample collection, demonstrating the feasibility of same-day public notification of current water quality conditions. SIGNIFICANCE AND IMPACT OF THE STUDY: This study presents the first report of in-situ qPCR enumeration of both faecal indicators and harmful algal species in coastal marine waters. We utilize a robotic device for in-situ quantification of enterococci, the human-specific marker in Bacteriodales and Pseudo-nitzschia spp. from the same water samples collected and processed in-situ. The results demonstrate that rapid, in-situ monitoring can be utilized to identify and quantify multiple health-relevant micro-organisms important in water quality monitoring and that this monitoring can be used to inform same-day notifications.

Assessment and genomic analysis of Salmonella and Campylobacter from different stages of an integrated no-antibiotics-ever (NAE) broiler complex: a longitudinal study.

The objective of this study was to determine prevalence and perform genomic analysis of Salmonella spp. and Campylobacter spp. isolated from different stages of an integrated NAE broiler complex. Environmental samples were screened with 3M-Molecular Detection System (MDS) and MDS positive samples were further processed for confirmation of results and identification. Core genome-based phylogenies were built for both bacteria isolated from this study along with selected NCBI genomes. The odds ratios and 95% confidence limits were compared among stages and sample types (α < 0.05) using multivariable model. Based on MDS results, 4% and 18% of total samples were positive for Salmonella spp. and Campylobacter spp. respectively. The odds of Salmonella detection in hatchery samples were 2.58 times as likely as compared to its detection in production farms' samples (P = 0.151) while the odds of Campylobacter detection in production farms' samples were 32.19 times as likely as its detection in hatchery (P = 0.0015). Similarly, the odds of Campylobacter detection in boot swabs, soil, water, and miscellaneous samples were statistically significant (P < 0.05) as compared with fly paper as reference group. The serovars identified for Salmonella were Typhimurium, Barranquilla, Liverpool, Kentucky, Enteritidis, Luciana, and Rough_O:r:1,5. For Campylobacter, the species identified were Campylobacter jejuni and Campylobacter coli. Phylogeny results show close genetic relatedness among bacterial strains isolated from different locations within the same stage and between different stages. The results show possibility of multiple entry points of such bacteria entering broiler complex and can potentially contaminate the final raw product in the processing plant. It suggests the need for a comprehensive control strategy with strict biosecurity measures and best management practices to minimize or eliminate such pathogens from the poultry food chain.

Synergistic activation of lamellar bismuth selenide anchored functionalized carbon nanofiber for detecting hazardous carbendazim in environmental water samples.

Pesticides pollute natural water reservoirs through persistent accumulation. Therefore, their toxicity and degradability are serious issues. Carbendazim (CBZ) is a pesticide used against fungal infections in agricultural crops, and its overexploitation detrimentally affects aquatic ecosystems and organisms. It is necessary to design a logical, efficient, and field-deployable method for monitoring the amount of CBZ in environmental samples. Herein, a nano-engineered bismuth selenide (Bi2Se3)/functionalized carbon nanofiber (f-CNF) nanocomposite was utilized as an electrocatalyst to fabricate an electrochemical sensing platform for CBZ. Bi2Se3/f-CNF exhibited a substantial electroactive surface area, high electrocatalytic activity, and high conductivity owing to the synergistic interaction of Bi2Se3 with f-CNF. The structural chemical compositions and morphology of the Bi2Se3/f-CNF nanocomposite were confirmed by X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), and field-emission scanning electron microscopy (FESEM). Electrochemical analysis was carried out using cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), and differential pulse voltammetry (DPV). The voltammetry and impedance experiments exposed that the Bi2Se3/f-CNF-modified GCE has attained adequate electrocatalytic function with amended features of electron transportation (Rct = 35.93 Ω) and improved reaction sites (0.082 cm2) accessible by CBZ moiety along with exemplary electrochemical stability (98.92%). The Bi2Se3/f-CNF nanocomposite exhibited higher sensitivity of 0.2974 µA µM-1cm-2 and a remarkably low limit of detection (LOD) of 1.04 nM at a broad linera range 0.001-100 µM. The practicability of the nanocomposite was tested in environmental (tap and pond water) samples, which supports excellent signal amplification with satisfactory recoveries. Hence, the Bi2Se3/f-CNF nanocomposite is a promising electrode modifier for detecting CBZ.

Development and optimization of sampling techniques for environmental samples from African swine fever virus-contaminated surfaces with no organic contaminants.

African swine fever (ASF) is a highly contagious diseases in domestic pigs and wild boars with up to 100% mortality. ASF virus (ASFV) is a causative agent responsible for ASF and highly resistant in environments, which creates a significant challenge for the control and eradication of the virus. Despite the geographical expansion of ASFV and international movement of products to sustain the swine production system, there is limited knowledge on the use of environmental samples to perform surveillance to prevent the introduction of ASFV into ASFV-free areas and for control of transmission in affected areas. Therefore, this study aimed to develop and optimize sampling techniques for environmental samples for ASFV detection. The stainless steel surfaces were contaminated with ASFV-infected blood, swabbed using different devices, and then processed through different techniques. The environmental samples were processed and tested using qPCR analysis. The results showed that the use of pre-moistened gauze surgical sponges, sweeping pads, and sponge sticks resulted in increased sensitivity, when compared to either dry sampling devices or Dacron swab. In particular, the combination of the sponge stick and the commercial nucleic acid preservative supported the best detection of ASFV DNA on the clean stainless steel surfaces evaluated. Pre-incubation for the short period of time and centrifugation at low speed were sufficient to provide satisfactory diagnostic sensitivity of ASFV detection using qPCR for environmental samples. Our findings contribute to the development of techniques for environmental samples for ASFV surveillance to prevent the introduction and dissemination of ASFV.