Beyond the Spectrum: Unleashing the Potential of Infrared Radiation in Poultry Industry Advancements.

The poultry industry is dynamically advancing production by focusing on nutrition, management practices, and technology to enhance productivity by improving feed conversion ratios, disease control, lighting management, and exploring antibiotic alternatives. Infrared (IR) radiation is utilized to improve the well-being of humans, animals, and poultry through various operations. IR radiation occurs via electromagnetic waves with wavelengths ranging from 760 to 10,000 nm. The biological applications of IR radiation are gaining significant attention and its utilization is expanding rapidly across multiple sectors. Various IR applications, such as IR heating, IR spectroscopy, IR thermography, IR beak trimming, and IR in computer vision, have proven to be beneficial in enhancing the well-being of humans, animals, and birds within mechanical systems. IR radiation offers a wide array of health benefits, including improved skin health, therapeutic effects, anticancer properties, wound healing capabilities, enhanced digestive and endothelial function, and improved mitochondrial function and gene expression. In the realm of poultry production, IR radiation has demonstrated numerous positive impacts, including enhanced growth performance, gut health, blood profiles, immunological response, food safety measures, economic advantages, the mitigation of hazardous gases, and improved heating systems. Despite the exceptional benefits of IR radiation, its applications in poultry production are still limited. This comprehensive review provides compelling evidence supporting the advantages of IR radiation and advocates for its wider adoption in poultry production practices.

Ultraspecific One-Pot CRISPR-Based "Green-Yellow-Red" Multiplex Detection Strategy Integrated with Portable Cartridge for Point-of-Care Diagnosis.

Versatile, informative, sensitive, and specific nucleic acid detection plays a crucial role in point-of-care pathogen testing, genotyping, and disease monitoring. In this study, we present a novel one-pot Cas12b-based method coupled with the "Green-Yellow-Red" strategy for multiplex detection. By integrating RT-LAMP amplification and Cas12b cleavage in a single tube, the entire detection process can be completed within 1 h. Our proposed method exhibits high specificity, enabling the discrimination of single-base mutations with detection sensitivity approaching single molecule levels. Additionally, the fluorescent results can be directly observed by the naked eye or automatically analyzed using our custom-designed software Result Analyzer. To realize point-of-care detection, we developed a portable cartridge capable of both heating and fluorescence excitation. In a clinical evaluation involving 20 potentially SARS-CoV-2-infected samples, our method achieved a 100% positive detection rate when compared to standard RT-PCR. Furthermore, the identification of SARS-CoV-2 variants using our method yielded results that were consistent with the sequencing results. Notably, our proposed method demonstrates excellent transferability, allowing for the simultaneous detection of various pathogens and the identification of mutations as low as 0.5% amidst a high background interference. These findings highlight the tremendous potential of our developed method for molecular diagnostics.

TriD-LAMP: A pump-free microfluidic chip for duplex droplet digital loop-mediated isothermal amplification analysis.

Digital nucleic acid amplification techniques are powerful and attractive approaches for providing sensitive and absolute quantification in biology. Among these, digital loop-mediated isothermal amplification (dLAMP) shows the potential for field detection, since its robustness and independence from thermal cycling. The key of dLAMP is to generate a large number of individual droplets or microwells. However, an auxiliary precision pump is always required for sample digitalization. In addition, current systems for droplet dLAMP usually need to transfer the droplets after digitalization or amplification. Herein, we developed and evaluated a pump-free microfluidic chip for duplex droplet dLAMP (TriD-LAMP) detection. This chip was designed based on step emulsification and contains a droplet generation zone and a droplet storage zone. Droplets are formed through the step due to the difference in Laplace pressure. There are 64 parallel nozzles that could generate tens of thousands of uniform droplets manually (variation <5%). The storage zone for droplets collection was previously filled with oil containing fluorosurfactant that keeps the droplets from fusing and evaporation during the amplification. Therefore, this custom chip is able to perform all stages of the dLAMP process without transferring droplets. Combined with the optimized fluorescent probe method, the chip achieves accurate quantification of the E. coli DNA down to 19.8 copies/µL. As a proof of concept, the simultaneous quantification of two targets was successfully realized on this custom chip. Conclusively, this integrated, pump-free TriD-LAMP chip can provide a promising tool for multiple targets detection in clinical diagnostics and point-of-care applications.

Two-dimensional nanocomposite-based electrochemical sensor for rapid determination of trans-resveratrol.

A two-dimensional nanocomposite-based disposable electrochemical sensor was fabricated for the rapid analysis of trans-resveratrol (TRA) in red wine. The sensor was prepared by modifying graphene-molybdenum disulfide (Gr-MoS2) nanocomposite on the surface of screen-printed electrode (SPE). Results show that the Gr-MoS2 nanocomposite with sheet-on-sheet structure can accelerate the oxidation reaction kinetics of TRA due to its large effective electrochemical surface area and high electron transfer rate. As a result, the Gr-MoS2 nanocomposite appears the synergistic effects, making the highly sensitive detection of TRA come true. The prepared sensor showed a linear response in TRA concentration from 1.0 to 200 µmol L-1 (with a limit of detection of 0.45 µmol L-1). After validating the accuracy with high performance liquid chromatography (HPLC), this nanocomposite-based electrochemical sensor can be applied for the detection of TRA in real red wine samples.

End-point dual specific detection of nucleic acids using CRISPR/Cas12a based portable biosensor.

A CRISPR/Cas12a based portable biosensor (Cas12a-PB) was developed to simultaneously visually detect CaMV35S promoter and Lectin gene from genetically modified (GM) soybean powders (Roundup Ready@). The Cas12a-PB, mainly made of polymethylmethacrylate (PMMA) and PMMA tape, has a connection structure, three channels and three detection chambers. The CRISPR/Cas12a detection reagents were preloaded in detection chambers and the reaction tube was connected to the connection structure by screw threads. After amplification, the amplicons were gone into three detection chambers by swinging the Cas12a-PB to conduct dual detection. Positive samples would produce green fluorescence while negative samples were black under the irradiation of 490 nm LED light. In this study, the Cas12a-PB successively combined with ordinary PCR, rapid PCR and loop-mediated isothermal amplification (LAMP) to achieve dual detection, which made detection process more convenient and portable. As low as 0.1% transgenic ingredients in soybean powders could be detected and the specificity of Cas12a-PB was confirmed with GM maize powders (MON810, GA21), GM soybean powders (DP305423), non-GM peanut and rice as targets. In the end, an amplification chamber combining with Cas12a-PB on a PMMA chip was further designed to eliminate the use of reaction tube and mineral oil, which made operation simpler. The established Cas12a-PB would provide a new reliable solution for multiple targets detection in clinic diagnostics, food safety, etc.

Magnetically separable and recyclable bamboo-like carbon nanotube-based FRET assay for sensitive and selective detection of Hg2.

The global occurrence of toxic hazards in aquatic ecosystems has aroused concern about the potential impacts on the ecological environment and human health in recent decades. Mercury(II) ions that originate from widespread sources including the mining industry, fossil fuel consumption, and industrial wastes are now well known as a highly toxic pollutant. Despite various detection methods which have been reported to sense Hg2+, it still poses a great challenge for us to develop a new effective sensing platform to replenish current fluorescent detection techniques. Here, we report a novel fluorescent biosensor using bamboo-like magnetic carbon nanotubes (BMCNTs) and FAM-labeled T-rich ssDNA for efficient detection of Hg2+ in aqueous solution. The proposed biosensor shows a good response toward Hg2+ detection over a linear response range of 0.05~1 µM (R2 = 0.98) with a detection limit of 20 nM. It also exhibits the capability to discriminate Hg2+ ions with negligible response to other metal ions, such as Ca2+, Cd2+, Cu2+, Mg2+, Mn2+, Ni2+, Pb2+, and Zn2+. Interestingly, the BMCNTs could be separated and recycled easily by using an external magnet, which means a much more cost-effective, easy-to-operate, and eco-friendly method for Hg2+ ion detection.

Rapid on-site detection of genetically modified soybean products by real-time loop-mediated isothermal amplification coupled with a designed portable amplifier.

Herein, we developed a rapid method for detection of genetically modified soybean (GTS 40-3-2) products using loop-mediated isothermal amplification (LAMP). A crude 5-minute extraction method was established for DNA extraction from soybeans and soybean products. LAMP reaction for CaMV35S promoter was optimized and the fastest threshold time (Tt) was 14 min with 4 mM magnesium ions at 63 °C. A portable instrument was designed to perform real-time LAMP in the field. As little as 0.1% GM soybean, specifically, or 0.5% GM ingredients in Chinese traditional tofu could be detected in 30 min from sampling to results. We used this method to further assess other five soybean products to determine whether they contained transgenic ingredients and compared the results with those obtained using PCR, which suggested the proposed method was applicable for rapid detection of genetically modified soybean in food products.

A fast and visual method for duplex shrimp pathogens detection with high specificity using rapid PCR and molecular beacon.

Molecular diagnosis of genome is one of the major methods for pathogens detection. The commonly used PCR method can realize an exponential amplification of the target gene but is time-consuming. In this work, we proposed a duplex and visual method using rapid PCR combined with molecular beacons to specifically detect two kinds of shrimp pathogens in one reaction tube. We only need to observe the fluorescence change of the reaction tube with naked eye to determine the result. A home-made automatic transfer equipment allows reaction tubes shuttling quickly between two water baths to achieve rapid PCR amplification. A simple device was also designed to present the detection results easily determined with naked eye. This duplex and visual detection method is fast, low-cost and of high specificity. From DNA extraction to results judgment, only 15 min was enough. Infectious Hypodermal and Hematopoietic Necrosis Virus (IHHNV) and Vibrio parahaemolyticus (VP) are two common shrimp pathogens which were chosen as our detection objects. This method may give a possibility to conduct end-point visual duplex detection, which may make a positive influence on the pathogen prevention.

An anti-passivation ink for the preparation of electrodes for use in electrochemical immunoassays.

p-Nitrophenylphosphate (PNPP) is usually employed as the substrate for enzyme-linked immunosorbent assays. p-Nitrophenol (PNP), the product of PNPP, with the catalyst alkaline phosphatase (ALP), will passivate an electrode, which limits applications in electrochemical analysis. A novel anti-passivation ink used in the preparation of a graphene/ionic liquid/chitosan composited (rGO/IL/Chi) electrode is proposed to solve the problem. The anti-passivation electrode was fabricated by directly writing the graphene-ionic liquid-chitosan composite on a single-side conductive gold strip. A glassy carbon electrode, a screen-printed electrode, and a graphene-chitosan composite-modified screen-printed electrode were investigated for comparison. Scanning electron microscopy was used to characterize the surface structure of the four different electrodes and cyclic voltammetry was carried out to compare their performance. The results showed that the rGO/IL/Chi electrode had the best performance according to its low peak potential and large peak current. Amperometric responses of the different electrodes to PNP proved that only the rGO/IL/Chi electrode was capable of anti-passivation. The detection of cardiac troponin I was used as a test example for electrochemical immunoassay. Differential pulse voltammetry was performed to detect cardiac troponin I and obtain a calibration curve. The limit of detection was 0.05 ng/ml.

Field Detection of Citrus Huanglongbing Associated with ' Candidatus Liberibacter Asiaticus' by Recombinese Polymerase Amplification within 15 min.

' Candidatus Liberibacter asiaticus' (Las) is the most prevalent bacterium associated with huanglongbing, which is one of the most destructive diseases of citrus. In this paper, an extremely rapid and simple method for field detection of Las from leaf samples, based on recombinase polymerase amplification (RPA), is described. Three RPA primer pairs were designed and evaluated. RPA amplification was optimized so that it could be accomplished within 10 min. In combination with DNA crude extraction by a 50-fold dilution after 1 min of grinding in 0.5 M sodium hydroxide and visual detection via fluorescent DNA dye (positive samples display obvious green fluorescence while negative samples remain colorless), the whole detection process can be accomplished within 15 min. The sensitivity and specificity of this RPA-based method were evaluated and were proven to be equal to those of real-time PCR. The reliability of this method was also verified by analyzing field samples.

Counting DNA molecules with visual segment-based readouts in minutes.

An ultrafast and extremely simple approach was proposed to count the number of DNA molecules without any microfluidic-based device. By directly counting the number of amplicon clusters in a capillary, the absolute amount of DNA molecules could be easily determined.

Rapid, Sensitive, and Carryover Contamination-Free Loop-Mediated Isothermal Amplification-Coupled Visual Detection Method for 'Candidatus Liberibacter asiaticus'.

Huanglongbing is a devastating citrus disease, and 'Candidatus Liberibacter asiaticus' (Las) is the most prevalent huanglongbing-associated bacterium. Its field detection remains challenging. In this work, a visual, rapid, sensitive, and carryover contamination-free method was developed for field detection of Las. Leaf samples were treated with 500 µL of 0.5 M sodium hydroxide solution for 3 min, and 50-fold dilutions were directly amplified by loop-mediated isothermal amplification. Then, a novel SYTO-9-based visual detection method was used to evaluate amplification results without uncapping operation. Negative samples remained colorless, while positive samples generated obvious green fluorescence, which could be easily distinguished by the naked eye with a mini-fluorescent-emission cartridge developed originally. The proposed detection method could be accomplished within 40 min and is about 100 times more sensitive than conventional TaqMan polymerase chain reaction. The reliability of this method was also verified by analyzing practical samples.

A Novel Impedimetric Microfluidic Analysis System for Transgenic Protein Cry1Ab Detection.

Impedimetric analysis method is an important tool for food safety detection. In this work, a novel impedimetric microfluidic analysis system consisted of a printed gold electrode chip and a microfluidic flow cell was developed for sensitive and selective detection of transgenic protein Cry1Ab. Anti-Cry1Ab aptamer coated magnetic beads were used to recognize transgenic protein Cry1Ab and form Cry1Ab-aptamer modified magnetic beads. After separation, the obtained Cry1Ab-aptamer modified magnetic beads were dissolved in 0.01 M mannitol and followed by injection into the microfluidic flow cell for impedimetric measurement. At the frequency of 358.3 Hz, the impedance signal shows a good linearity with the concentrations of Cry1Ab protein at a range from 0 to 0.2 nM, and the detection limit is 0.015 nM. The results demonstrate that the impedimetric microfluidic analysis system provides an alternative way to enable sensitive, rapid and specific detection of transgenic protein Cry1Ab.

Design and Elementary Evaluation of a Highly-Automated Fluorescence-Based Instrument System for On-Site Detection of Food-Borne Pathogens.

A simple, highly-automated instrument system used for on-site detection of foodborne pathogens based on fluorescence was designed, fabricated, and preliminarily tested in this paper. A corresponding method has been proved effective in our previous studies. This system utilizes a light-emitting diode (LED) to excite fluorescent labels and a spectrometer to record the fluorescence signal from samples. A rotation stage for positioning and switching samples was innovatively designed for high-throughput detection, ten at most in one single run. We also developed software based on LabVIEW for data receiving, processing, and the control of the whole system. In the test of using a pure quantum dot (QD) solution as a standard sample, detection results from this home-made system were highly-relevant with that from a well-commercialized product and even slightly better reproducibility was found. And in the test of three typical kinds of food-borne pathogens, fluorescence signals recorded by this system are highly proportional to the variation of the sample concentration, with a satisfied limit of detection (LOD) (nearly 10²-10³ CFU·mL-1 in food samples). Additionally, this instrument system is low-cost and easy-to-use, showing a promising potential for on-site rapid detection of food-borne pathogens.

Terahertz sensing of chlorpyrifos-methyl using metamaterials.

By squeezing electromagnetic energy into small volumes near a metal-dielectric interface, plasmonics provide many routes to enhance and manipulate light-matter interactions, which presents new strategies for signal enhancing technologies. As an extension of the ideas of plasmonics to the terahertz (THz) range, metamaterials have shown great potential in sensing applications. In this study, terahertz time-domain spectroscopy (THz-TDS) combined with metamaterials was used to detect chlorpyrifos-methyl (CM), which is one type of the broad-spectrum organophosphorus pesticides. The results demonstrate that sensitivity is greatly improved using THz metamaterials, with the limit of detection (LOD) of CM reaching 0.204mgL-1, which is lower than the World Health Organization's provisional guideline limit for CM in vegetables (1mgL-1). The results indicated that THz spectroscopy combined with metamaterials could be a valuable method for highly sensitive THz applications, presenting a new strategy for food quality and safety control in the future.

Discrimination of Transgenic Rice containing the Cry1Ab Protein using Terahertz Spectroscopy and Chemometrics.

Spectroscopic techniques combined with chemometrics methods have proven to be effective tools for the discrimination of objects with similar properties. In this work, terahertz time-domain spectroscopy (THz-TDS) combined with discriminate analysis (DA) and principal component analysis (PCA) with derivative pretreatments was performed to differentiate transgenic rice (Hua Hui 1, containing the Cry1Ab protein) from its parent (Ming Hui 63). Both rice samples and the Cry1Ab protein were ground and pressed into pellets for terahertz (THz) measurements. The resulting time-domain spectra were transformed into frequency-domain spectra, and then, the transmittances of the rice and Cry1Ab protein were calculated. By applying the first derivative of the THz spectra in conjunction with the DA model, the discrimination of transgenic from non-transgenic rice was possible with accuracies up to 89.4% and 85.0% for the calibration set and validation set, respectively. The results indicated that THz spectroscopic techniques and chemometrics methods could be new feasible ways to differentiate transgenic rice.

Development of an aptamer-based impedimetric bioassay using microfluidic system and magnetic separation for protein detection.

An aptamer-based impedimetric bioassay using the microfluidic system and magnetic separation was developed for the sensitive and rapid detection of protein. The microfluidic impedance device was fabricated through integrating the gold interdigitated array microelectrode into a flow cell made of polydimethylsiloxane (PDMS). Aptamer modified magnetic beads were used to capture and separate the target protein, and concentrated into a suitable volume. Then the complexes were injected into the microfluidic flow cell for impedance measurement. To demonstrate the high performance of this novel detection system, thrombin was employed as the target protein. The results showed that the impedance signals at the frequency of 90 kHz have a good linearity with the concentrations of thrombin in a range from 0.1 nM to 10nM and the detection limit is 0.01 nM. Compared with the reported impedimetric aptasensors for thrombin detection, this method possesses several advantages, such as the increasing sensitivity, improving reproducibility, reducing sample volume and assay time. All these demonstrate the proposed detection system is an alternative way to enable sensitive, rapid and specific detection of protein.

Impedimetric immunosensor based on gold nanoparticles modified graphene paper for label-free detection of Escherichia coli O157:H7.

In this study, a low-cost and robust impedimetric immunosensor based on gold nanoparticles modified free-standing graphene paper electrode for rapid and sensitive detection of Escherichia coli O157:H7 (E. coli O157:H7) was developed. Graphene paper was prepared by chemical reduction of graphene oxide paper obtained from vacuum filtration method. Scanning electron microscope, Raman spectroscopy and X-ray diffraction techniques were employed to investigate the surface morphology and crystal structure of the prepared graphene paper. The gold nanoparticles were grown on the surface of graphene paper electrode by one-step electrodeposition technique. The immobilization of anti-E. coli O157:H7 antibodies on paper electrode were performed via biotin-streptavidin system. Electrochemical impedance spectroscopy was used to detect E. coli O157:H7 captured on the paper electrode. Results show that the developed paper immunosensor possesses greatly enhanced sensing performance, such as wide linear range (1.5 × 10(2)-1.5 × 10(7) cfu mL(-1)), low detection limit (1.5 × 10(2) cfu mL(-1)), and excellent specificity. Furthermore, flexible test demonstrate the graphene paper based sensing device has high tolerability to mechanical stress. The strategy of structurally integrating metal nanomaterials, graphene paper, and biorecognition molecules would provide new insight into design of flexible immunosensors for routine sensing applications.

Monitoring of Escherichia coli O157:H7 in food samples using lectin based surface plasmon resonance biosensor.

A novel surface plasmon resonance (SPR) biosensor using lectin as bioreceptor was developed for the rapid detection of Escherichia coli (E. coli) O157:H7. The selective interaction of lectins with carbohydrate components from bacterial cells surface was used as the recognition principle for the detection. Five types of lectins from Triticum vulgaris, Canavailia ensiformis, Ulex europaeus, Arachis hypogaea, and Maackia amurensis, were employed to evaluate the selectivity of the approach for binding E. coli O157:H7 effectively. A detection limit of 3×10(3) cfu mL(-1) was obtained for determination of E. coli O157:H7 when used the lectin from T. vulgaris as the binding molecule. Furthermore, the proposed biosensor was used to detect E. coli O157:H7 in real food samples. Results showed that the lectin based SPR biosensor was sensitive, reliable and effective for detection of E. coli O157:H7, which hold a great promise in food safety analysis.

New trends in impedimetric biosensors for the detection of foodborne pathogenic bacteria.

The development of a rapid, sensitive, specific method for the foodborne pathogenic bacteria detection is of great importance to ensure food safety and security. In recent years impedimetric biosensors which integrate biological recognition technology and impedance have gained widespread application in the field of bacteria detection. This paper presents an overview on the progress and application of impedimetric biosensors for detection of foodborne pathogenic bacteria, particularly the new trends in the past few years, including the new specific bio-recognition elements such as bacteriophage and lectin, the use of nanomaterials and microfluidics techniques. The applications of these new materials or techniques have provided unprecedented opportunities for the development of high-performance impedance bacteria biosensors. The significant developments of impedimetric biosensors for bacteria detection in the last five years have been reviewed according to the classification of with or without specific bio-recognition element. In addition, some microfluidics systems, which were used in the construction of impedimetric biosensors to improve analytical performance, are introduced in this review.