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1.
J Dairy Sci ; 107(5): 2748-2759, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38101746

RESUMO

A novel ratiometric electrochemical aptasensor based on split aptamer and Au-reduced graphene oxide (Au-rGO) nanomaterials was proposed to detect aflatoxin M1 (AFM1). In this work, Au-rGO nanomaterials were coated on the electrode through the electrodeposition method to increase the aptamer enrichment. We split the aptamer of AFM1 into 2 sequences (S1 and S2), where S1 was immobilized on the electrode due to the Au-S bond, and S2 was tagged with methylene blue (MB) and acted as a response signal. A complementary strand to S1 (CS1) labeled with ferrocene (Fc) was introduced as another reporter. In the presence of AFM1, CS1 was released from the electrode surface due to the formation of the S1-AFM1-S2 complex, leading to a decrease in Fc and an increase in the MB signal. The developed ratiometric aptasensor exhibited a linear range of 0.03 µg L-1 to 2.00 µg L-1, with a detection limit of 0.015 µg L-1 for AFM1 detection. The ratiometric aptasensor also showed a linear relationship from 0.2 µg L-1 to 1.00 µg L-1, with a detection limit of 0.05 µg L-1 in natural milk after sample pretreatment, indicating the successful application of the developed ratiometric aptasensor. Our proposed strategy provides a new way to construct aptasensors with high sensitivity and selectivity.


Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Compostos Ferrosos , Grafite , Metalocenos , Animais , Aflatoxina M1/análise , Aptâmeros de Nucleotídeos/química , Grafite/química , Técnicas Biossensoriais/métodos , Técnicas Biossensoriais/veterinária , Técnicas Eletroquímicas/métodos , Técnicas Eletroquímicas/veterinária , Limite de Detecção
2.
J Dairy Sci ; 105(7): 5545-5560, 2022 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-35534270

RESUMO

The non-systematic evolution of ligands by the exponential enrichment (non-SELEX) method was used in the present study for the selection of ß-casomorphin-7 (BCM-7)-specific aptamers. These aptamers were tested to evaluate their ability to detect BCM-7 peptide in the human urine sample. The method did not employ aptamer amplification and counterselection as used in conventional SELEX but included a negative round of selection. The selection was performed in a single day, and after 5 rounds, a total of 16 numbers of aptamer were identified through Sanger sequencing. Newly selected aptamers named sequence ID no. 3 have performed better than other aptamers in detecting the BCM-7 peptide. Sequence ID no. 3 was also compared with previously selected aptamers through the SELEX method and its performance was found to be better than old aptamers. The sensing experiment was tried on different platforms from magnetic beads to the membrane. In each strategy, satisfactory results were obtained with aptamers that recognized BCM-7 spiked in a human urine sample at a very low amount. The non-SELEX method is an easy and time-saving process for aptamer selection. Selection of viable aptamers from a large pool of sequences for sensing experiments is a tedious job; however, an attempt has been made to select aptamers on the basis of In Silico (http://www.unafold.org/, https://bioinformatics.ramapo.edu/QGRS/index.php) information, observing DNA band intensity on agarose gel and colorimetric results obtained on magnetic beads and membrane. These aptamers have the potential in biosensor making for detecting BCM-7 peptide in urine samples of autistic patients.


Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Animais , Aptâmeros de Nucleotídeos/genética , Técnicas Biossensoriais/métodos , Técnicas Biossensoriais/veterinária , Endorfinas , Humanos , Ligantes , Técnica de Seleção de Aptâmeros/métodos , Técnica de Seleção de Aptâmeros/veterinária
3.
J Dairy Sci ; 105(4): 2895-2907, 2022 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-35181133

RESUMO

Salmonella, as a common foodborne pathogen in dairy products, poses a great threat to human health. We studied a new detection method based on quantum dots (QD). A fluorescent biosensor with multiple fluorescent signal amplification based on a streptavidin (SA) biotin system and the polyamino linear polymer poly-l-lysine (PLL) were established to detect Salmonella in milk. First, Salmonella was captured on a black 96-well plate with paired Salmonella mAb to form a double-antibody sandwich. Second, SA was immobilized on biotin-modified mAb by SA-biotin specific bond. Then, the biotin-modified polylysine (BT-PLL) was bound on SA and specifically bonded again through the SA-biotin system. Finally, water-soluble CdSe/ZnS QD-labeled SA was added to a black 96-well plate for covalent coupling with BT-PLL. The fluorescent signal was amplified in a dendritic manner by the layer-by-layer overlap of SA and biotin and the covalent coupling of biotinylated PLL. Under optimal conditions, the detection limit was 4.9 × 103 cfu/mL in PBS. The detection limit was 10 times better than that of the conventional sandwich ELISA. In addition, the proposed biosensor was well specific and could be used for detecting Salmonella in milk samples.


Assuntos
Técnicas Biossensoriais , Pontos Quânticos , Animais , Técnicas Biossensoriais/veterinária , Biotina/química , Leite , Polilisina , Salmonella , Estreptavidina/química
4.
J Dairy Sci ; 105(3): 2108-2118, 2022 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-34998563

RESUMO

Foods contaminated by foodborne pathogens have always been a great threat to human life. Herein, we constructed an electrochemical immunosensor for Salmonella detection by using a Fe3O4@graphene modified electrode. Because of the excellent electrical conductivity and mechanical stability of graphene and the large specific surface area of Fe3O4, the Fe3O4@graphene nanocomposite exhibits an excellent electrical signal, which greatly increased the sensitivity of the immunosensor. Gold nanoparticles were deposited on Fe3O4@graphene nanocomposite by electrochemical technology for the immobilization of the antibody. Cyclic voltammetry was selected to electrochemically characterize the construction process of immunosensors. The microstructure and morphology of related nanocomposites were analyzed by scanning electron microscopy. Under optimized experimental conditions, a good linear relationship was achieved in the Salmonella concentration range of 2.4 × 102 to 2.4 × 107 cfu/mL, and the limit of detection of the immunosensor was 2.4 × 102 cfu/mL. Additionally, the constructed immunosensor exhibited acceptable selectivity, reproducibility, and stability and provides a new reference for detecting pathogenic bacteria in milk.


Assuntos
Técnicas Biossensoriais , Grafite , Nanopartículas Metálicas , Nanocompostos , Animais , Técnicas Biossensoriais/veterinária , Carbono , Técnicas Eletroquímicas/veterinária , Eletrodos , Ouro/química , Grafite/química , Imunoensaio/veterinária , Limite de Detecção , Nanopartículas Metálicas/química , Leite , Nanocompostos/química , Reprodutibilidade dos Testes , Salmonella
5.
J Dairy Sci ; 105(3): 1966-1977, 2022 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-34955267

RESUMO

Aflatoxin M1 (AFM1) is a common toxin in dairy products that causes acute and chronic human health disorders. Thus, the development of a rapid and accurate AFM1 detection method is of vital importance for food safety monitoring. This work was to develop a novel electrochemical aptasensor for sensitive and specific determination of AFM1. The dendritic-like nanostructure was formed on the gold electrode surface by layer-by-layer assembly of gold-silver core-shell nanoparticles modified with DNA conjugates. In the presence of AFM1, the specific recognition between AFM1 and Apt caused the disassociation of the DNA controlled dual Au@Ag conjugates from the surface of the electrode, causing less methylene blue to bind to the surface and weakening the electrochemical signal. The more AFM1 there is, the weaker the electrochemical signal. Transmission electron microscope results showed that the successfully synthesized Au@Ag nanoparticles exhibited a core-shell structure with Au as core and Ag as shell, and their average diameter was about 30 nm. Under optimal conditions, the electrochemical aptasensor showed a wide detection ranging from 0.05 ng mL-1 to 200 ng mL-1, and a low detection limit of 0.02 ng mL-1. Moreover, the proposed strategy has been successfully applied to the detection of AFM1 in cow, goat, and sheep milk samples with satisfactory recoveries ranging from 91.10% to 104.05%. This work can provide a novel rapid detection method for AFM1, and also provide a new sensing platform for the detection of other toxins.


Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Nanopartículas Metálicas , Aflatoxina M1/análise , Animais , Aptâmeros de Nucleotídeos/análise , Aptâmeros de Nucleotídeos/química , Técnicas Biossensoriais/métodos , Técnicas Biossensoriais/veterinária , DNA/análise , Limite de Detecção , Nanopartículas Metálicas/química , Leite/química , Ovinos , Prata
6.
Vet Parasitol ; 301: 109638, 2022 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-34920304

RESUMO

Canine visceral leishmaniasis (CVL) is the most aggressive and lethal form of leishmaniasis manifesting in dogs and represents a major public health concern. Although there are sufficiently sensitive molecular tools for CVL diagnosis, they are not accessible at the main points of disease dissemination, in which context serodiagnosis has been used as an alternative tool on the epidemiological control. As an attempt to develop more accurate immunodiagnostic assays, many antigens have been tested over the years, on different platforms. This review aimed to access studies reporting new antigens that can be applied for CVL serodiagnosis. Articles published from January of 2016 to March of 2021 were retrieved from Google Scholar, Science Direct, and PubMed, using "Canine Visceral Leishmaniasis" and "Serodiagnosis" as keywords. In total, 1527 articles were identified, of which 42 were selected based on exclusion factors. Sensitivity, specificity, sample size, and sample quality data were extracted by manual curation and analyzed. Of the selected articles, 26 contemplated ELISA, which enabled a more thorough comparison and a critical review of these studies. Soluble Leishmania Antigens (SLA) and the A2 protein were used as controls in 53.8 and 46.15 % of these articles, respectively, and were evaluated separately; their frequent use was questioned. Subsequently, articles that evaluated other assay platforms, such as immunochromatography, immunosensors, and others, were also reported and evaluated. Finally, data relative to validation studies of commercial kits were briefly discussed. Our results show that there are several antigens with great potential for the development of accurate diagnostic tools, but further testing is required. The critical analysis also brings insights that can be useful for more assertive diagnostic development of more robust tools for CVL serodiagnosis.


Assuntos
Técnicas Biossensoriais , Doenças do Cão , Leishmania infantum , Leishmaniose Visceral , Animais , Antígenos de Protozoários , Técnicas Biossensoriais/veterinária , Doenças do Cão/diagnóstico , Cães , Ensaio de Imunoadsorção Enzimática/veterinária , Imunoensaio/veterinária , Leishmaniose Visceral/diagnóstico , Leishmaniose Visceral/veterinária , Sensibilidade e Especificidade , Testes Sorológicos/veterinária
7.
Comp Immunol Microbiol Infect Dis ; 79: 101708, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34481108

RESUMO

This study reports development of a novel point of care assay, namely an enhanced immuno-dot blot assay, for discrimination of anti-Toxoplasma IgG and anti-Toxoplasma IgM antibodies. This method has been designed based on formation of a sandwich complex between a gold nanoprobe (chitosan gold nanoparticle-anti-human IgG or anti-IgM) and anti- Toxoplasma lysate antigen (TLA) which holds anti-TLA antibodies, either IgG or IgM. Briefly, anti-human IgG or anti-IgM antibody was conjugated to chitosan gold nanoparticles via glutaraldehyde chemistry. Then, lysate antigen was immobilized on the surface of nitrocellulose membrane, which followed by addition of the sera sample and gold nanoprobes. The positive signals were readily detectable via observation with naked eye. This positive color change was further intensified via gold enhancement chemistry. The intensity of biosensor signal was proportional to the concentration of active antibodies on the surface of nanoparticles, titer of T. gondii antibodies in the sera samples and concentration of Toxoplasma lysate antigen coated on the nitrocellulose membrane. A minimum concentration to use the antibodies for conjugation, to detect titer of Toxoplasma IgG and IgM antibodies, and the concentration of TLA coated in nitrocellulose membrane were 0.5 mg/mL, 2 IU/mL, 10 IU/mL, and 20 µg/mL, respectively. This enhanced immuno-dot blot assay offers a simple diagnostic technique without expensive equipment requirement for distinguishing of anti- T. gondii IgM and IgG antibodies in field conditions, pregnant women, and immunocompromised patients.


Assuntos
Técnicas Biossensoriais , Nanopartículas Metálicas , Toxoplasma , Toxoplasmose , Animais , Anticorpos Antiprotozoários , Técnicas Biossensoriais/veterinária , Colorimetria/veterinária , Ensaio de Imunoadsorção Enzimática/veterinária , Feminino , Ouro , Imunoensaio/veterinária , Imunoglobulina M , Gravidez , Toxoplasmose/diagnóstico
8.
J Dairy Sci ; 104(12): 12295-12302, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34538487

RESUMO

Salmonella is a foodborne pathogen that has contributed to numerous food safety accidents worldwide, making it necessary to detect contamination at an early stage. A pair of specific primers based on the invA gene of Salmonella was designed for PCR. Target double-stranded DNA (dsDNA) from PCR was purified and denatured at high temperature to obtain target single-stranded DNA (ssDNA). Two carboxyfluorescein-labeled hairpin probes (H1-FAM and H2-FAM) were designed with complementary portions to the ssDNA sequence so that binding could trigger H1-FAM and H2-FAM hybridization chain reaction (HCR) to produce a long dsDNA complex. In this study, graphene oxide (GO) was used in the development of a homogeneous fluorescence detection platform for Salmonella. Using this HCR-GO assay platform, Salmonella detection was completed in 3.5 h. Salmonella was reliably and specifically detected with a limit of detection (LOD) of 4.2 × 101 cfu/mL in pure culture. Moreover, this new HCR-GO assay platform was successfully applied to the detection of Salmonella in artificially contaminated milk with a LOD of 4.2 × 102 cfu/mL.


Assuntos
Técnicas Biossensoriais , Grafite , Animais , Técnicas Biossensoriais/veterinária , Leite , Salmonella/genética
9.
J Dairy Sci ; 104(11): 11486-11498, 2021 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-34454766

RESUMO

Rapid and sensitive detection of foodborne pathogens is of great importance for food safety. Here, a set of nuclear magnetic resonance (NMR) biosensors based on a O-carboxymethyl chitosan target gadolinium (Gd) probe was developed to quickly detect Salmonella in milk by combining NMR technology and bioimmunotechnology with membrane filtration technology. First, O-carboxymethyl chitosan (O-CMC) was biotinylated to prepare biotinylated O-carboxymethyl chitosan (biotin-O-CMC) through amide reaction, and biotinylated magnetic complexes (biotin-O-CMC-Gd) were obtained by using O-CMC, which has strong chelating adsorption on Gd. The target probe was obtained by combining biotin-O-CMC-Gd with the biotinylated antibody (biotin-antibody) via streptavidin (SA) by introducing the SA-biotin system. Then, Salmonella was captured by the target probe through antigen-antibody interaction. Finally, NMR was used to measure the longitudinal relaxation time (T1) of the filtrate collected by membrane filtration. This NMR biosensor with good specificity and high efficiency can detect Salmonella with the sensitivity of 1.8 × 103 cfu/mL within 2 h; in addition, it can realize the detection of complex samples because of its strong anti-interference capability and may open up a new method for rapid detection of Salmonella, which has a great application potential.


Assuntos
Técnicas Biossensoriais , Biotina , Animais , Técnicas Biossensoriais/veterinária , Quitosana/análogos & derivados , Gadolínio , Espectroscopia de Ressonância Magnética , Leite , Salmonella , Estreptavidina
10.
Arch Virol ; 166(10): 2763-2778, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34342747

RESUMO

White spot syndrome virus (WSSV) is a significant threat to the aquaculture sector, causing mortality among crabs and shrimps. Currently available diagnostic tests for WSSV are not rapid or cost-effective, and a new detection method is therefore needed. This study demonstrates the development of a biosensor by functionalization of magnetosomes with VP28-specific antibodies to detect WSSV in seafood. The magnetosomes (1 and 2 mg/ml) were conjugated with VP28 antibody (0.025-10 ng/µl), as confirmed by spectroscopy. The magnetosome-antibody conjugate was used to detect the VP28 antigen. The binding of antigen to the magnetosome-antibody complex resulted in a change in absorbance. The magnetosome-antibody-antigen complex was then concentrated and brought near a screen-printed carbon electrode by applying an external magnetic field, and the antigen concentration was determined using impedance measurements. The VP28 antigen (0.025 ng/µl) bound more efficiently to the magnetosome-VP28 antibody complex (0.025 ng/µl) than to the VP28 antibody (0.1 ng/µl) alone. The same assay was repeated to detect the VP28 antigen (0.01 ng/µl) in WSSV-infected seafood samples using the magnetosome-VP28 antibody complex (0.025 ng/µl). The WSSV in the seafood sample was also drawn toward the electrode due to the action of magnetosomes controlled by the external magnetic field and detected using impedance measurement. The presence of WSSV in seafood samples was verified by Western blot and RT-PCR. Cross-reactivity assays with other viruses confirmed the specificity of the magnetosome-based biosensor. The results indicate that the use of the magnetosome-based biosensor is a sensitive, specific, and rapid way to detect WSSV in seafood samples.


Assuntos
Técnicas Biossensoriais/veterinária , Magnetossomos , Alimentos Marinhos/virologia , Vírus da Síndrome da Mancha Branca 1/isolamento & purificação , Animais , Anticorpos Antivirais/química , Anticorpos Antivirais/imunologia , Aquicultura , Reações Cruzadas , Espectroscopia Dielétrica , Ensaio de Imunoadsorção Enzimática , Microbiologia de Alimentos , Magnetossomos/química , Magnetossomos/imunologia , Penaeidae/virologia , Reprodutibilidade dos Testes , Proteínas do Envelope Viral/análise , Proteínas do Envelope Viral/imunologia , Vírus da Síndrome da Mancha Branca 1/imunologia
11.
Philos Trans R Soc Lond B Biol Sci ; 376(1830): 20200218, 2021 08 02.
Artigo em Inglês | MEDLINE | ID: mdl-34121461

RESUMO

Farmed aquatic animals represent an increasingly important source of food for a growing human population. However, the aquaculture industry faces several challenges with regard to producing a profitable, ethical and environmentally sustainable product, which are exacerbated by the ongoing intensification of operations and increasingly extreme and unpredictable climate conditions. Fortunately, bio-sensors capable of measuring a range of environmental, behavioural and physiological variables (e.g. temperature, dissolved gases, depth, acceleration, ventilation, heart rate, blood flow, glucose and l-lactic acid) represent exciting and innovative tools for assessing the health and welfare of farmed animals in aquaculture. Here, we illustrate how these state-of-the-art technologies can provide unique insights into variables pertaining to the inner workings of the animal to elucidate animal-environment interactions throughout the production cycle, as well as to provide insights on how farmed animals perceive and respond to environmental and anthropogenic perturbations. Using examples based on current challenges (i.e. sub-optimal feeding strategies, sub-optimal animal welfare and environmental changes), we discuss how bio-sensors can contribute towards optimizing the growth, health and welfare of farmed animals under dynamically changing on-farm conditions. While bio-sensors currently represent tools that are primarily used for research, the continuing development and refinement of these technologies may eventually allow farmers to use real-time environmental and physiological data from their stock as 'early warning systems' and/or for refining day-to-day operations to ethically and sustainably optimize production. This article is part of the theme issue 'Measuring physiology in free-living animals (Part I)'.


Assuntos
Bem-Estar do Animal , Aquicultura/instrumentação , Técnicas Biossensoriais/veterinária , Peixes/fisiologia , Tecnologia de Sensoriamento Remoto/veterinária , Animais , Animais Domésticos/fisiologia , Técnicas Biossensoriais/instrumentação , Tecnologia de Sensoriamento Remoto/instrumentação , Tecnologia/instrumentação
12.
J Dairy Sci ; 104(8): 8506-8516, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-34053767

RESUMO

Effective testing tools for Escherichia coli O157:H7 can prevent outbreaks of foodborne illness. In this paper, a smartphone-based colorimetric aptasensor was developed using functionalized gold nanoparticles (GNP) and multi-walled carbon nanotubes (MWCNT) for monitoring E. coli O157:H7 in milk. The maximum absorption peak of GNP bonded with aptamer (Apt) generated evident transformation from 518 to 524 nm. The excess GNP-Apt was removed by functionalized MWCNT magnetized with carbonyl iron powder (CIP) and hybridized with a DNA probe, whereas the GNP-Apt immobilized on E. coli O157:H7 remained in the system. In the presence of a high-salt solution, the GNP-Apt that captured E. coli O157:H7 remained red, but the free GNP-Apt aggregated and appeared blue. The chromogenic results were analyzed by a smartphone-based colorimetric device that was fabricated using acrylic plates, a light-emitting diode, and a mobile power pack. To our knowledge, this was the first attempt to use a smartphone-based colorimetric aptasensor employing the capture of GNP-Apt coupled with separation of MWCNT@CIP probe to detect E. coli O157:H7. The aptasensor exhibited good reproducibility and no cross-reaction for other bacteria. A concentration of 8.43 × 103 cfu/mL of E. coli O157:H7 could be tested in pure culture, and 5.24 × 102 cfu/mL of E. coli O157:H7 could be detected in artificially contaminated milk after 1 h of incubation. Therefore, the smartphone-based colorimetric aptasensor was an efficient tool for the detection of E. coli O157:H7 in milk.


Assuntos
Técnicas Biossensoriais , Escherichia coli O157 , Nanopartículas Metálicas , Nanotubos de Carbono , Animais , Técnicas Biossensoriais/veterinária , Colorimetria/veterinária , Microbiologia de Alimentos , Ouro , Leite , Reprodutibilidade dos Testes , Smartphone
13.
J Dairy Sci ; 104(6): 6944-6960, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-33814150

RESUMO

The aim of this study was to develop and evaluate a method for detecting Mycobacterium avium ssp. paratuberculosis (MAP) bacteria in bovine fecal, milk, and colostrum samples using immunomagnetic beads (IMB) and a rhodamine hydrazone immunosensor. Immunomagnetic beads were prepared by using purified antibodies from hyperimmunized sera that were coupled to Fe nanoparticles with diethylene triamine pentaacetic acid (DTPA) or ethyl (dimethyl aminopropyl) carbodiimide (EDC)-N-hydroxy succinimide (NHS) as linkers. Rhodamine hydrazone particles were synthesized and coupled to IgY anti-MAP antibodies using DTPA or EDC-NHS linkers. Separation efficiency of the IMB was tested on bovine fecal, milk, and colostrum samples experimentally contaminated with MAP. The studied methods were evaluated on their ability to detect MAP and separate bacteria in complex mediums. The ELISA results indicated 95% efficacy in antibody coupling to IMB, with the DTPA-IMB method being more efficient than the EDC-NHS-IMB method. By using the DTPA-IMB method, MAP bacteria were successfully recovered from fecal, milk, and colostrum samples. The DTPA-IMB method used in combination with the rhodamine hydrazone immunosensor had a limit of detection equal to 30 and 30,000 MAP cells/mL using chromogenic and fluorescent properties, respectively. Combining the DTPA-IMB separation method with the rhodamine hydrazone immunosensor provides a fast, sensitive, and cost-beneficial method for detecting MAP in bovine feces, milk, and colostrum.


Assuntos
Técnicas Biossensoriais , Doenças dos Bovinos , Mycobacterium avium subsp. paratuberculosis , Paratuberculose , Animais , Técnicas Biossensoriais/veterinária , Bovinos , Doenças dos Bovinos/diagnóstico , Colostro , Fezes , Feminino , Hidrazinas , Imunoensaio/veterinária , Leite , Gravidez , Rodaminas
15.
J Vet Sci ; 21(5): e79, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-33016024

RESUMO

BACKGROUND: Recently, the pork industry of Thailand faced an epidemic of highly virulent strains of porcine reproductive and respiratory syndrome virus (PRRSV), which spread throughout Southeast Asia, including the Lao People's Democratic Republic and Cambodia. Hence, the rapid and on-site screening of infected pigs on a farm is essential. OBJECTIVES: To develop the new aptamer as a biosensor for detection PRRSV which are rapid and on-site screening of infected pig. METHODS: New aptamers against PRSSV were identified using the combined techniques of capillary electrophoresis, colorimetric assay by gold nanoparticles, and quartz crystal microbalance (QCM). RESULTS: Thirty-six candidate aptamers of the PRRSV were identified from the systematic evolution of ligands by exponential enrichment (SELEX) by capillary electrophoresis. Only 8 out of 36 aptamers could bind to the PRSSV, as shown in a colorimetric assay. Of the 8 aptamers tested, only the 1F aptamer could bind specifically to the PRSSV when presented with the classical swine fever virus and a pseudo rabies virus. The QCM was used to confirm the specificity and sensitivity of the 1F aptamer with a detection limit of 1.87 × 1010 particles. CONCLUSIONS: SELEX screening of the aptamer equipped with capillary electrophoresis potentially revealed promising candidates for detecting the PRRSV. The 1F aptamer exhibited the highest specificity and selectivity against the PRRSV. These findings suggest that 1F is a promising aptamer for further developing a novel PRRSV rapid detection kit.


Assuntos
Aptâmeros de Nucleotídeos/química , Técnicas Biossensoriais/veterinária , Síndrome Respiratória e Reprodutiva Suína/diagnóstico , Vírus da Síndrome Respiratória e Reprodutiva Suína/isolamento & purificação , Técnica de Seleção de Aptâmeros/veterinária , Animais , Técnicas Biossensoriais/métodos , Colorimetria/veterinária , Ouro/química , Nanopartículas Metálicas/química , Síndrome Respiratória e Reprodutiva Suína/virologia , Técnicas de Microbalança de Cristal de Quartzo/veterinária , Técnica de Seleção de Aptâmeros/métodos , Sensibilidade e Especificidade , Suínos
16.
J Fish Biol ; 97(6): 1644-1650, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-32889736

RESUMO

Metabolic rates are one of many measures that are used to explain species' response to environmental change. Static respirometry is used to calculate the standard metabolic rate (SMR) of fish, and when combined with exhaustive chase protocols it can be used to measure maximum metabolic rate (MMR) and aerobic scope (AS) as well. While these methods have been tested in comparison to swim tunnels and chambers with circular currents, they have not been tested in comparison with a no-chase control. We used a repeated-measures design to compare estimates of SMR, MMR and AS in European perch Perca fluviatilis following three protocols: (a) a no-chase control; (b) a 3-min exhaustive chase; and (c) a 3-min exhaustive chase followed by 1-min air exposure. We found that, contrary to expectations, exhaustive chase protocols underestimate MMR and AS at 18°C, compared to the no-chase control. This suggests that metabolic rates of other species with similar locomotorty modes or lifestyles could be similarly underestimated using chase protocols. These underestimates have implications for studies examining metabolic performance and responses to climate change scenarios. To prevent underestimates, future experiments measuring metabolic rates should include a pilot with a no-chase control or, when appropriate, an adjusted methodology in which trials end with the exhaustive chase instead of beginning with it.


Assuntos
Técnicas Biossensoriais/veterinária , Consumo de Oxigênio/fisiologia , Percas/metabolismo , Animais , Técnicas Biossensoriais/instrumentação , Técnicas Biossensoriais/métodos , Natação/fisiologia
17.
J Dairy Sci ; 103(8): 7585-7597, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32505394

RESUMO

Commonly used lactose assays [enzymatic spectrophotometric absorbance (EZA) and HPLC] for dairy ingredients are relatively expensive and time consuming. A blood glucose meter (BGM)-based method has successfully been documented as a rapid lactose assay in milk. However, the BGM-based method has not been evaluated in dairy ingredients. The objective of this study was to evaluate the BGM-based lactose analysis method in whey-derived (WD) and skim milk-derived (SMD) ingredients. The study was carried out in 4 phases. In phase 1, the effect of pH and lactose concentrations on the BGM reading was investigated using a factorial design with 2 factors: pH (6.02-7.50) and lactose (0.2 or 0.4%). We found that BGM readings were significantly affected by lower pH values at both lactose levels. In phase 2, the effect of total solids and ingredient type was investigated using a factorial design with 2 factors: ingredient type (WD or SMD) and total solids (0-8%). It was observed that the BGM reading was significantly affected by ingredient type and total solids. Phase 3 involved developing a linear relationship between the BGM reading and the EZA reference method to ascertain the accuracy of the proposed BGM method. Different ingredient types (WD or SMD) and non-lactose solids (0.5-27%) model ingredient dilutions prepared over a range of lactose contents (0.08-0.62%) were measured using the BGM and EZA methods. The average absolute percentage bias difference between the BGM method and EZA reference method results for these model dilutions was found to be between 2.2 and 7.3%. In phase 4, 15 samples procured from commercial sources ranging from 0.01 to 81.9% lactose were evaluated using the BGM method and EZA reference method. The average absolute percentage bias difference for lactose results between the 2 methods ranged from 3.6 to 5.0% and 5.3 to 9.7% for well-performing and poorly performing meters, respectively. Overall, the BGM method is a promising tool for rapid and low-cost analysis of lactose in both high-lactose and low-lactose dairy ingredients.


Assuntos
Técnicas Biossensoriais/veterinária , Laticínios/análise , Lactose/análise , Animais , Glicemia , Bovinos , Leite/química , Soro do Leite/química , Proteínas do Soro do Leite/química
18.
Sensors (Basel) ; 20(6)2020 Mar 18.
Artigo em Inglês | MEDLINE | ID: mdl-32197444

RESUMO

The recent trend in electrocardiogram (ECG) device development is towards wireless body sensors applied for patient monitoring. The ultimate goal is to develop a multi-functional body sensor that will provide synchronized vital bio-signs of the monitored user. In this paper, we present an ECG sensor for long-term monitoring, which measures the surface potential difference between proximal electrodes near the heart, called differential ECG lead or differential lead, in short. The sensor has been certified as a class IIa medical device and is available on the market under the trademark Savvy ECG. An improvement from the user's perspective-immediate access to the measured data-is also implemented into the design. With appropriate placement of the device on the chest, a very clear distinction of all electrocardiographic waves can be achieved, allowing for ECG recording of high quality, sufficient for medical analysis. Experimental results that elucidate the measurements from a differential lead regarding sensors' position, the impact of artifacts, and potential diagnostic value, are shown. We demonstrate the sensors' potential by presenting results from its various areas of application: medicine, sports, veterinary, and some new fields of investigation, like hearth rate variability biofeedback assessment and biometric authentication.


Assuntos
Técnicas Biossensoriais/instrumentação , Eletrocardiografia/instrumentação , Frequência Cardíaca/fisiologia , Monitorização Fisiológica/instrumentação , Telemedicina , Animais , Identificação Biométrica/instrumentação , Identificação Biométrica/métodos , Técnicas Biossensoriais/métodos , Técnicas Biossensoriais/veterinária , Cardiotocografia/instrumentação , Eletrocardiografia/métodos , Eletrocardiografia/veterinária , Eletrodos/veterinária , Desenho de Equipamento , Feminino , Cavalos , Humanos , Aplicativos Móveis , Monitorização Ambulatorial/instrumentação , Monitorização Ambulatorial/métodos , Monitorização Ambulatorial/veterinária , Monitorização Fisiológica/métodos , Monitorização Fisiológica/veterinária , Valor Preditivo dos Testes , Gravidez , Cuidado Pré-Natal/métodos , Processamento de Sinais Assistido por Computador/instrumentação , Medicina Esportiva/instrumentação , Medicina Esportiva/métodos , Telemedicina/instrumentação , Telemedicina/métodos , Telemetria/instrumentação , Telemetria/métodos , Telemetria/veterinária , Fatores de Tempo , Medicina Veterinária/instrumentação , Medicina Veterinária/métodos , Tecnologia sem Fio/instrumentação
19.
Animal ; 14(S1): s187-s195, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-32024562

RESUMO

In this position paper, I shall summarise the current status of sensor technologies in ruminant livestock farming with emphasis on dairy cattle, outline the case for why I believe that sensor technologies could revolutionise global dairy farming in a positive way, describe the significant barriers that exist if that goal is to be achieved and highlight the benefits to animal wellbeing, profitability and sustainability that could result if the technologies are implemented to a significant extent. I shall not provide a comprehensive review of the sensor technology literature since that has been done before, but I intend to provide a sensible amount of background information and data that will allow the reader to obtain a picture not only of today's sensor usage but, more importantly, the possible future direction of dairy animal-oriented sensor technologies, and I shall substantiate my claims and conclusions with relevant literature.


Assuntos
Bem-Estar do Animal , Técnicas Biossensoriais/veterinária , Bovinos/fisiologia , Monitores de Aptidão Física/veterinária , Ruminantes/fisiologia , Agricultura , Animais , Indústria de Laticínios , Feminino , Gado
20.
Vet Rec ; 185(18): 572, 2019 11 09.
Artigo em Inglês | MEDLINE | ID: mdl-31554712

RESUMO

BACKGROUND: Lameness is a major health, welfare and production-limiting condition for the livestock industries. The current 'gold-standard' method of assessing lameness by visual locomotion scoring is subjective and time consuming, whereas recent technological advancements have enabled the development of alternative and more objective methods for its detection. METHODS: This study evaluated a novel lameness detection method using micro-Doppler radar signatures to categorise animals as lame or non-lame. Animals were visually scored by veterinarian and radar data were collected for the same animals. RESULTS: A machine learning algorithm was developed to interpret the radar signatures and provide automatic classification of the animals. Using veterinary scoring as a standard method, the classification by radar signature provided 85 per cent sensitivity and 81 per cent specificity for cattle and 96 per cent sensitivity and 94 per cent specificity for sheep. CONCLUSION: This radar sensing method shows promise for the development of a highly functional, rapid and reliable recognition tool of lame animals, which could be integrated into automatic, on-farm systems for sheep and cattle.


Assuntos
Técnicas Biossensoriais/veterinária , Doenças dos Bovinos/diagnóstico , Coxeadura Animal/diagnóstico , Radar , Doenças dos Ovinos/diagnóstico , Algoritmos , Animais , Bovinos , Aprendizado de Máquina , Sensibilidade e Especificidade , Ovinos
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