Thesis (Ph. D.)--University of Texas at Austin, 2007. / Vita. Includes bibliographical references.

Αντικείμενο της διπλωματικής αυτής εργασίας είναι η μελέτη ενός ZigBee ασύρματου δικτύου βιοαισθητήρων και η δυνατότητα διασύνδεσης του με μεγαλύτερα δίκτυα όπως το Ίντερνετ. Αρχικά γίνεται μια αναφορά στην χρησιμοποίηση δικτύων τηλεϊατρικής, στη σπουδαιότητά τους για τον άνθρωπο καθώς και σε ολοκληρωμένες λύσεις που έχουν προταθεί. Έπειτα παρουσιάζονται και περιγράφονται βιοαισθητήρες που χρησιμοποιούνται σε ένα WBAN όπως και η λειτουργία τους. Στη συνέχεια γίνεται εκτεταμένη ανάλυση όλων των επιπέδων του πρωτοκόλλου Zigbee/802.15.4 στοχεύοντας πιο συγκεκριμένα στη ζώνη μετάδοσης των 2,4 GHz και το πως αυτή μπορεί να χρησιμοποιηθεί κρατώντας υψηλό το QOS , απαραίτητη προϋπόθεση ενός δικτύου τηλεϊατρικής . Τέλος μελετώνται οι πιθανοί τρόποι διασύνδεσης δικτύων αισθητήρων και γίνεται προσομοίωση ενός TCP/IP δικτύου στο οποίο καταλήγουν τα δεδομένα τα οποία συγκεντρώνονται από μια σειρά αισθητήρων με σκοπό τη μελέτη της δυνατότητας χρήσης ενός τέτοιου συστήματος. / Subject of this diploma work is to study a ZigBee wireless biosensor network and connectivity with other networks like the internet. Initially is made a reference to the use of telemedicine networks, the importance for man as well as integrated solutions that have been proposed. Then there are presented and described biosensors used in a WBAN as their function. Then is made an analysis of all levels of the protocol Zigbee/802.15.4 targeting the particular zone of 2,4 GHz and how it can be used while holding a high QOS. Finally are examined possible ways of interconnecting networks of sensors and is made a simulation of a TCP / IP where the collected data from the sensors end.

Submitted by Johnny Rodrigues (johnnyrodrigues@ufcg.edu.br) on 2017-11-21T21:46:23Z No. of bitstreams: 1 Hugo Yves Cordeiro Eulálio - Dissertação PPG-CEMat - 2016..pdf: 9966635 bytes, checksum: 5ea265c58dad0db100d0e913e7338764 (MD5) / Made available in DSpace on 2017-11-21T21:46:23Z (GMT). No. of bitstreams: 1 Hugo Yves Cordeiro Eulálio - Dissertação PPG-CEMat - 2016..pdf: 9966635 bytes, checksum: 5ea265c58dad0db100d0e913e7338764 (MD5) Previous issue date: 2016-02-29 / Um dos desafios da atualidade está em encontrar métodos simples, rápidos e seletivos para determinações específicas de compostos variados nas mais diversas aplicações. Um componente fundamental para os biossensores, tratase do transdutor, responsável por transformar e conduzir os sinais gerados devido as reações entre analito alvo e biorreceptor. Em testes iniciais viu-se que a deposição se soltava facilmente da superfície do eletrodo então com o intuito de melhorar a condutividade dos eletrodos foi realizado eletrodeposições do composto Sb/Sb2O3 nas superfícies dos suportes metálicos e visando uma melhor adesão do eletrodepositado nos metais foram realizadas eletrocorrosões nestes suportes com solução de HCl. O objetivo deste trabalho foi analisar o comportamento de suportes metálicos para transdutores, submetidos a eletrodeposição de antimônio e de óxido de antimônio (Sb/Sb2O3) e ao processo de eletrocorrosão ácida, para o emprego em biossensores. Com a finalidade de identificar qual composto estava sendo depositado no metal, para visualizar a morfologia e o recobrimento e, ainda, confirmar essa possível melhor adesão realizou-se caracterizações como: microscopia eletrônica de varredura, difração de raios x, análise gravimétrica das amostras, análise da rugosidade superficial e testes de adesividade do depositado. A partir dos difratogramas foi possível observar a eletrodeposição do Sb metálico na superfície metálica. As imagens obtidas por MEV mostraram morfologias distintas que variaram dependendo do metal utilizado, da metodologia utilizada, porém todos apresentaram uma característica pulverulenta, pois são facilmente arrancados da superfície. O teste de gravimetria acusou que o cobre é mais sensível a corrosão que o aço inox. O teste de adesão mostrou que na média o cobre obteve uma maior adesão, porém a amostra que teve maior adesão foi utilizando uma fita de aço. Conclui-se que dentre os dois suportes metálicos escolhidos, levando em consideração apenas o tratamento superficial e a metodologia utilizada, o aço inox é o melhor suporte a ser empregado. / One of nowadays' challenges is to find simple, fast and selective methods for specific determinations of various compounds in various applications. A key component to the biosensor, is the transducer, responsible for transforming and lead the signals generated due to reactions between the target analyte and biorreceptor. In initial tests it was seen that the deposition easily let go of the electrode surface then, to improve the conductivity, they were conducted electrodeposition compound Sb / Sb2O3 on the surfaces of metal brackets and to better adhesion of the electroplated in metals were performed electroerosions on these stands with HCl solution. The aim of this study was to analyze the metal brackets behavior for transducers subjected to electrodeposition of antimony and antimony oxide (Sb / Sb2O3) and acid electroerosion process, for use in biosensors. In order to identify which compound was being deposited on the metal, to see the morphology and the coating and also confirm this possible better adhesion some characterizations were done, as scanning electron microscopy, X-ray diffraction, gravimetric analysis of the samples, analysis of surface roughness and deposited adhesion tests. From the XRD patterns was observed only the electrodeposition of Sb metallic on the metal surface. The images obtained by SEM showed distinct morphologies that varied depending on the metal used, the methodology used, but all had a powdery characteristic, because they are easily torn from the surface. The gravimetric test showed that copper is more sensitive to corrosion than stainless steel. The adhesion test showed that the average copper obtained an increased adhesion, but the sample had adhesion was greater using a steel tape. It is concluded that among the two metal supports chosen, taking into account only the surface treatment and the methodology used, the stainless steel is the best support to be used.

Made available in DSpace on 2016-05-17T16:51:15Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-07-29. Added 1 bitstream(s) on 2016-05-17T16:54:48Z : No. of bitstreams: 1 000863271.pdf: 3961818 bytes, checksum: ac858d20df08483eb5bdbe31474f4806 (MD5) / A espectroscopia de Raman intensificada por superfície (SERS, termo em inglês Surfaceenhanced Raman Spectroscopy) é uma técnica promissora que mostra a sensibilidade para a detecção da interação de biomoléculas que são importantes para detecção precoce de doenças. O vírus da imunodeficiência humana (HIV) têm sido um grande problema por várias décadas. Existem vários métodos de deteção baseados na interação específica de anticorpos, tais como, o ELISA e os testes rápidos (TR's). No entanto, novas estratégias têm sido desenvolvidas para rápido diagnóstico do vírus HIV, e uma prova de conceito de detecção do peptídeo p17-1 foi descrito neste trabalho. A proteina matriz p17 é uma essencial proteína no ciclo de replicação do vírus HIV. As fases iniciais da replicação do vírus envolve a pré integração do complexo do DNA no núcleo do p17 desempenhando um papel na ligação de RNA viral e transporte para a membrana. Neste trabalho foram descritos duas plataformas SERS para a detecção do vírus HIV baseado no peptide p17 -1 (sequência LSGGELDRWEKIRLPGG). O anticorpo foi imobilizado em um substrato de ouro usando duas diferentes camadas automontadas (SAM). A primeira SAM, os substratos de ouro foram imersos em uma solução aquosa de 11 mercaptoundecanóico (MUA). Na segunda SAM, os substratos foram imersos em uma mistura aquosa de politietileno glicol (SHPEG- COOH e SH-PEG-CH3). Aqui serão chamados de SAM-MUA e SAM-PEG, respectivamente. Ambas as SAM's foram imersas emu ma solução de anticorpo (anti-p17) e foram descritas como plataforma d captura MUA e PEG. Ambas plataformas foram funcionalizadas com o peptídeo p17-1. Sondas SERS foram preparadas com nanopartículas de ouro e revestidas com uma molécula Raman reporter (azul de Nilo A) e funcionalizadas com um anticorpo anti-p17. Estas estruturas (sonda SERS e plataformas de captura) formam um ensaio sanduíche... / Surface-enhanced Raman Scattering (SERS) technique offers great promises for simplified and sensitive detection of biomolecular interactions that are relevant for early disease diagnostics. Human immunodeficiency virus (HIV) has been a problem for decades. There are several methods of diagnostics based on antibodies specific reactions, such as enzyme-linked immunosorbent assays (ELISAs) and rapid test (RT). However, new strategies have been developed for rapid HIV diagnostics and, as a proof-of-concept, peptide p17-1 was considered here. The matrix protein p17 is a structural protein that is essential in the life cycle of the retrovirus The early stages of the virus replication involve the pre integration of the DNA complex into the nucleus P17 plays a role in RNA viral binding and transport to the membrane. Here were describe two new SERS platform for HIV detection based on peptide p17-1 (sequence LSGGELDRWEKIRLPGG). The antibody anti-p17 was immobilized in a planar gold surface using two differents self-assembled (SAM) techniques. First SAM, were obtained by immersion of the surface into ethanolic solution of 11-Mercaptoundecanoic acid (MUA). Second SAM were obtained by immersion in aqueous solution aquous mixtures of (SH-PEG-COOH/SH-PEG-CH3) and polyethylene glycol (PEG,). Here were describe the two platforms as SAM-MUA and SAMPEG, respectively. Both SAM's were immersed in a solution containing the anti-p17. Samples at this step were called capture platform-MUA and capture platform-PEG. Both capture platforms were funcionalizated with the peptide p17-1. SERS probes were prepared with gold nanoparticles coated with a Raman reporter molecule (Nile Blue A) and, functionalized with an anti-p17. These structures (SERS probe and capture platforms) allow for a sandwich assay, a strategy regularly used for high-sensitivity detection. The light blue color in the SERS mapping represents peptide strong...

Made available in DSpace on 2015-03-03T11:52:58Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-08-19Bitstream added on 2015-03-03T12:06:47Z : No. of bitstreams: 1 000810546_20160801.pdf: 1088702 bytes, checksum: cd367dff4dec1ca6d3501d00abf21caa (MD5) Bitstreams deleted on 2016-08-01T11:30:00Z: 000810546_20160801.pdf,. Added 1 bitstream(s) on 2016-08-01T11:30:56Z : No. of bitstreams: 1 000810546.pdf: 2492702 bytes, checksum: b28dd413d4faa07c29c95b215497fd1a (MD5) / Os corantes azo estão entre os mais utilizados pela indústria têxtil brasileira, representando de 20-40% dos corantes empregados para tingir algodão, rayon, nylon, seda, lã e couro. Um fator preocupante relacionado a estas substâncias é que estudos utilizando microorganismos e células de mamíferos têm demonstrado que diversos corantes azo apresentam atividade genotóxica, mesmo em baixas concentrações. Além disso, atualmente ainda não estão vigorando métodos oficiais para efetiva remoção dos corantes presentes nos efluentes gerados no processo de tingimento dos tecidos, e isso faz com que essas substâncias possam chegar à agua destinada ao consumo, e assim, o desenvolvimento de dispositivos sensores para o monitoramento dos corantes em água se torna de grande importância. Diante disso, o presente trabalho versa sobre o desenvolvimento de um imunossensor impedimétrico para detecção e quantificação do corante azo Disperso Red 1 (DR1) em baixos níveis de concentração em água tratada e para tal, duas estratégias metodológicas foram estudadas. A primeira delas se fundamentou na construção do imunossensor empregando eletrodos impressos de ouro modificados com monocamadas auto-organizadas. Monocamadas tioladas de cistamina, ácido lipóico e p-aminotiofenol foram estudadas para promoção do acoplamento dos anticorpos sobre a superfície do eletrodo. Caracterizações realizadas por voltametria cíclica (VC) e espectroscopia de impedância eletroquímica (EIS) demonstraram a efetiva imobilização dos anticorpos sobre a superfície dos eletrodos impressos, com aumento sucessivo do recobrimento e no valor de resistência de transferência de carga (Rct), respectivamente. No entanto, para todas as rotas estudadas encontraram-se dificuldades em se obter adequada repetibilidade entre as diferentes medidas e estabilidade das etapas de modificação, além de não ser observada... / Azo dyes are among the most used by Brazilian textile industry, representing 20-40% of dyes used for dyeing cotton, rayon, nylon, silk, wool and leather. A worrying factor related to these substances is that studies using micro-organisms and mammalian cells have shown that many azo dyes have genotoxic activity, even at low concentrations. Besides, currently there are not oficial methods for effective removal of dyes present in the effluents generated in the dyeing process, and this leads these substances can reach the water for consumption, and thus the development of sensor devices for monitoring the dyes in water becomes of great importance. Therefore, this project is focused on development of an impedimetric immunosensor for detection and quantification of the azo dye Disperse Red 1 (DR1) at low concentration levels in treated water and for this, two methodological strategies were studied. The first one was based on the construction of the immunosensor employing gold printed electrodes modified with self-assembled monolayers. Thiolated monolayers of cystamine, lipoic acid and p-aminothiophenol were studied to promote the coupling of the antibodies on the electrode surface. Characterizations performed by cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) demonstrated the effective immobilization of antibodies on the surface of the printed electrodes, with successive increase in the surface coverage and in the charge transference resistance (Rct), respectively. However for all ways investigated, difficulties in obtaining adequate repeatability between measures of stability and modification steps were observed, as well a non-linear relationship between the values of ?Rct and the concentration of antigen DR1, which was due by the heterogeneity of the surface of different printed electrodes. Thus, the second strategy employed glassy carbon electrode for immunosensor construction...

abstract: The past two decades have been monumental in the advancement of microchips designed for a diverse range of medical applications and bio-analysis. Owing to the remarkable progress in micro-fabrication technology, complex chemical and electro-mechanical features can now be integrated into chip-scale devices for use in biosensing and physiological measurements. Some of these devices have made enormous contributions in the study of complex biochemical processes occurring at the molecular and cellular levels while others overcame the challenges of replicating various functions of human organs as implant systems. This thesis presents test data and analysis of two such systems. First, an ISFET based pH sensor is characterized for its performance in a continuous pH monitoring application. Many of the basic properties of ISFETs including I-V characteristics, pH sensitivity and more importantly, its long term drift behavior have been investigated. A new theory based on frequent switching of electric field across the gate oxide to decrease the rate of current drift has been successfully implemented with the help of an automated data acquisition and switching system. The system was further tested for a range of duty cycles in order to accurately determine the minimum length of time required to fully reset the drift. Second, a microfluidic based vestibular implant system was tested for its underlying characteristics as a light sensor. A computer controlled tilt platform was then implemented to further test its sensitivity to inclinations and thus it‟s more important role as a tilt sensor. The sensor operates through means of optoelectronics and relies on the signals generated from photodiode arrays as a result of light being incident on them. ISFET results show a significant drop in the overall drift and good linear characteristics. The drift was seen to reset at less than an hour. The photodiodes show ideal I-V comparison between photoconductive and photovoltaic modes of operation with maximum responsivity at 400nm and a shunt resistance of 394 MΩ. Additionally, post-processing of the tilt sensor to incorporate the sensing fluids is outlined. Based on several test and fabrication results, a possible method of sealing the open cavity of the chip using a UV curable epoxy has been discussed. / Dissertation/Thesis / M.S. Electrical Engineering 2011

>Magister Scientiae - MSc / The tuberculosis (TB) disease to this day remains one of the world’s prominent killerdiseases. Pyrazinamide (PZA) is one of the most commonly prescribed anti- tuberculosis (anti-TB) drugs due to its ability to significantly shorten the TB treatment period from the former nine months to the current six months duration. However, excess PZA in the body causes hepatotoxicity and damages the liver. This hepatotoxicity, together with the resistance of the bacteria to treatment drugs, poor medication and inappropriate dosing, greatly contribute to the high incidents of TB deaths and diseases that are due to side effects (such as liver damage). This brings about the calls for alternative methods for ensuring reliable dosing of the drug, which will be specific from person to person due to inter-individual differences in drug metabolism. A novel biosensor system for monitoring the metabolism of PZA was prepared with a Ni-PPI-PPy star copolymer and cytochrome P450 2E1 (CYP2E1) deposited onto a platinum electrode. The nanobiosensor system exhibited enhanced electro-activity that is attributed to the catalytic effect of the incorporated star copolymer. The biosensor had a sensitivity of 0.142 µA.nM-1, and a dynamic linear range (DLR) of 0.01 nM-0.12 nM (1.231 – 7.386 ng/L PZA). The limit of detection of the biosensor was found to be 0.00114 nM (0.14 ng/L) PZA. From the HPLC peakconcentration (Cmax) of PZA determined 2 h after drug intake is 2.79 – 3.22 ng.L-1,which is very detectable with the nanobiosensor as it falls within the dynamic linear range.

The aim of the thesis was to identify and hence investigate the physical properties of liquid crystals that influence their potential as components of biosensor devices. Silicon surfaces presenting photolithographically fabricated arrays of 50nm thick gold spots were used as the model for a biosensor that detects the surface binding of a biological analyte. The spots ranged in diameter from 2μm to 16μm and their spatial separation varied between 5μm to 40μm. A Self Assembled Monolayer (SAM) of the thiol 3-mercaoptopropionic acid was used to control the surface chemistry of the gold. The responses of the nematic liquid crystals 5CB, E7, ZLI 1695, ZLI 1132 and MDA 01-2012 to were measured by optical microscopy. The spots were seen to induce a tilted planar alignment in the liquid crystals in their nematic phase for spot diameters down to 4μm and for all separations. Anchoring transitions between different tilt angles were observed between spots for some arrays. This was linked to a change in anchoring energy at the gold, possibly stemming from the angle of gold deposition. When heated through the nematic to isotropic phase transition cross defects were observed to nucleate on the gold spots for all spot sizes above 4μm. On cooling through the transition grid patterns of defects were observed to nucleate pinned between the spots for arrays of spots with length scales between 10μm and 20μm. The birefringence and elastic constants K11 and K33 of the liquid crystals were measured for temperatures up to their nematic to isotropic transition points. The birefringences of the liquid crystals at the transition were found to range between 0.003 and 0.007. The device thickness was varied between 7μm and 40μm. Values for the elastic constants were found to range between 1pN and 4pN. The intensity of monochromatic light (670nm) reflected from the arrays as the liquid crystals were cooled through the phase transition was found to increase for smaller values of the elastic constants and found to be highest where the grid of defects on the array was observed most clearly. The effect on the intensity of the birefringence and cell thickness was shown to be small compared to the effect of elasticity. Two possible biosensor designs are proposed. The first would identify the presence of a biological analyte at a surface by the change in alignment of a liquid crystal. This type of sensor would be optimised by carefully controlling the anchoring energy of the liquid crystal at the surface to minimise the quantity of surface binding required to induce an anchoring transition. The second would detect the presence at a patterned surface of an analyte by the defects that form over the pattern as the liquid crystal changes between the nematic to isotropic phases. This type of sensor would be optimised by choosing a liquid crystal with small elastic constants at the phase transition and by designing a patterned surface with length scales between 10microns and 20microns.

abstract: Engineered nanoporous substrates made using materials such as silicon nitride or silica have been demonstrated to work as particle counters or as hosts for nano-lipid bilayer membrane formation. These mechanically fabricated porous structures have thicknesses of several hundred nanometers up to several micrometers to ensure mechanical stability of the membrane. However, it is desirable to have a three-dimensional structure to ensure increased mechanical stability. In this study, circular silica shells used from Coscinodiscus wailesii, a species of diatoms (unicellular marine algae) were immobilized on a silicon chip with a micrometer-sized aperture using a UV curable polyurethane adhesive. The current conducted by a single nanopore of 40 nm diameter and 50 nm length, during the translocation of a 27 nm polystyrene sphere was simulated using COMSOL multiphysics and tested experimentally. The current conducted by a single 40 nm diameter nanopore of the diatom shell during the translocation of a 27 nm polystyrene sphere was simulated using COMSOL Multiphysics (28.36 pA) and was compared to the experimental measurement (28.69 pA) and Coulter Counting theory (29.95 pA).In addition, a mobility of 1.11 x 10-8 m2s-1V-1 for the 27 nm polystyrene spheres was used to convert the simulated current from spatial dependence to time dependence. To achieve a sensing diameter of 1-2 nanometers, the diatom shells were used as substrates to perform ion-channel reconstitution experiments. The immobilized diatom shell was functionalized using silane chemistry and lipid bilayer membranes were formed. Functionalization of the diatom shell surface improves bilayer formation probability from 1 out of 10 to 10 out of 10 as monitored by impedance spectroscopy. Self-insertion of outer membrane protein OmpF of E.Coli into the lipid membranes could be confirmed using single channel recordings, indicating that nano-BLMs had formed which allow for fully functional porin activity. The results indicate that biogenic silica nanoporous substrates can be simulated using a simplified two dimensional geometry to predict the current when a nanoparticle translocates through a single aperture. With their tiered three-dimensional structure, diatom shells can be used in to form nano-lipid bilayer membranes and can be used in ion-channel reconstitution experiments similar to synthetic nanoporous membranes. / Dissertation/Thesis / Doctoral Dissertation Electrical Engineering 2015

Submitted by Aelson Maciera (aelsoncm@terra.com.br) on 2017-05-11T17:29:15Z No. of bitstreams: 1 TeseTAN.pdf: 1864712 bytes, checksum: 75782f61cc6526aa709949101798707e (MD5) / Approved for entry into archive by Ronildo Prado (ronisp@ufscar.br) on 2017-05-18T20:22:16Z (GMT) No. of bitstreams: 1 TeseTAN.pdf: 1864712 bytes, checksum: 75782f61cc6526aa709949101798707e (MD5) / Approved for entry into archive by Ronildo Prado (ronisp@ufscar.br) on 2017-05-18T20:22:25Z (GMT) No. of bitstreams: 1 TeseTAN.pdf: 1864712 bytes, checksum: 75782f61cc6526aa709949101798707e (MD5) / Made available in DSpace on 2017-05-22T20:17:57Z (GMT). No. of bitstreams: 1 TeseTAN.pdf: 1864712 bytes, checksum: 75782f61cc6526aa709949101798707e (MD5) Previous issue date: 2016-06-29 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Bovine babesiosis and anaplasmosis are distinct diseases, which constitute the syndrome called tick fever, characterized by infection of red blood cells. Its distribution in herds causes the reduction in productivity and high economic losses to livestock worldwide. Anaplasma marginale, Babesia bovis, and Babesia bigemina are the predominant species in Brazil. Rhipicephalus (Boophilus) microplus is the biological vector of babesiosis and anaplasmosis may be transmitted by ticks, hematophagous flies, and fomites. It is known that the water buffalo can become infected with Babesia and Anaplasma, but little is known the extent of the infection and how it affects the health of these animals. Thus, the present study aimed to determine the frequency and the level of infection by B. bovis, B. bigemina and A. marginale in 108 water buffalo (50 calves and 58 adult females), naturally infested with the tick R. (B). microplus raised in farms located in areas of endemic stability in São Paulo state. From each animal, a blood sample from the jugular vein was taken for DNA extraction and packed cell volume (PCV) determination. Samples from auricular vessels were taken for blood smears. The body temperature was measured with a mercury thermometer column. Electrochemical impedance spectroscopy technique was used to differentiate the serum of uninfected animals from the serum of animals infected with B. bovis. To this end, antigens derived from culture were immobilized on gold electrodes (150 nm thick) to give a biosensor device using the compound [Fe (CN) 6] 3- / 4-proof redox. The peripheral blood smears were stained with May-Grunwald-Giemsa for research of hemoparasites by optical microscopy. DNA extractions were performed using the Easy DNA TM Kit (Invitrogen). DNA amplification protocols were tested with primers specific to B. bigemina, B. bovis, and A. marginale using nested PCR (nPCR) and quantitative real-time PCR (qPCR). qPCR was used to estimate the number of copies of the gene cytochrome b (mt-cytB) of both babesias and Anaplasma msp 1b gene in all samples. Merozoites of B. bigemina were seen in blood smears of three calves from the Alambari herd (all less than 0.1 of parasitemia). Molecular techniques, nPCR, and qPCR, were more sensitive in the detection of parasites that direct examination of the blood smears and the frequencies of infection were 20:37% and 100% for B. bovis and 59.26% and 100% to B. bigemina, respectively. CN of the mt-cytB gene of B. bovis and B. bigemina showed significant effects (p <0.05) of herd age, species, and their interaction. The CN values were higher (p≤0.05) for B. bovis (2.81 ± 0:07) when compared to B. bigemina (2.61 ± 12:07) and A. marginale (0:57 ± 0:07). These data suggest a high frequency of infection by B. bovis and B. bigemina in the population of water buffalo studied. Preliminary testing of diagnosing infection with B. bovis device showed changes in the impedance in the system used and clearly demonstrated that the biosensor can detect infected animals, which can be exploited for rapid detection of B. bovis infections and also extended for the test to other parasites. / A babesiose e anaplasmose bovina são enfermidades distintas, que constituem o complexo chamado de Tristeza Parasitária Bovina (TPB), caracterizada por infecção das células vermelhas do sangue. Sua distribuição nos rebanhos ocasiona redução na produtividade e grandes perdas econômicas para a pecuária mundial. Anaplasma marginale, Babesia bovis e Babesia bigemina são as espécies prevalentes no Brasil. O carrapato Rhipicephalus (Boophilus) microplus é o vetor biológico das babesioses, e a anaplasmose, além do carrapato pode ser transmitida por moscas hematófogas ou fômites. Sabe-se que, os búfalos podem se infectar com os agentes da TPB, porém pouco é conhecido a extensão dessa infecção e como ela afeta a saúde desses animais. Assim o presente estudo teve o objetivo de determinar a frequência e o nível de infecção por B. bovis, B. bigemina e A. marginale em 108 búfalos d’água (50 bezerros e 58 fêmeas adultas), naturalmente infestados com o carrapato R. (B). microplus, oriundos de fazendas localizadas em áreas endêmicas para as babesioses no estado de São Paulo. Foram colhidas amostras de sangue dos capilares auriculares para confeccção de esfregaços em lâminas de vidro e sangue da veia caudal para extração de DNA e determinação do volume globular (VG) pela técnica do microhematócrito. A técnica de espectroscopia de impedância eletroquímica foi realizada com o soro dos bovinos, com o objetivo de diferenciar animais saudáveis dos animais infectados com B. bovis. Para tanto, antígenos provenientes de cultura foram imobilizados em eletrodos de ouro (150 nm de espessura) dando origem a um dispositivo biossensor utilizando o composto [Fe (CN)6]3-/4-como prova redox. Os esfregaços de sangue periférico foram corados com May Grunwald-Giemsa para a pesquisa dos hemoparasitas por meio de microscopia óptica. As extrações de DNA foram feitas usando o Easy-DNATM Kit (Invitrogen). Foram testados protocolos de amplificação do DNA com primers específicos para B. bigemina, B. bovis e A. marginale por meio de Nested PCR (nPCR) e PCR em tempo real quantitativo (qPCR), usado para estimar o número de cópias (NC) do gene do citocromo b (mt-cytB) de ambas as babesias e do gene de superfície 1b (msp1b) de Anaplasma em todas as amostras. Foram visualizados merozoítas de B. bigemina nos esfregaços sanguíneos de três bezerros do rebanho de Alambari (todos com menos de 0,1 de parasitemia). As técnicas moleculares, nPCR e qPCR foram mais sensíveis para a detecção dos parasitas que o exame direto das lâminas, sendo que as frequências de infecção foram de 20.37% e 100% para B. bovis e 59.26% e 100% para B. bigemina, respectivamente. O NC do gene do mt-cytB de B. bovis e B. bigemina mostrou efeitos significativos (p<0.05) para rebanho-idade, espécies e sua interação. Os valores de NC foram superiores (p≤0.05) para B. bovis (2.81 ± 0.07) quando comparado a B. bigemina (2.61 ± 0.07) e A. marginale (0.57 ± 0.07). Estes dados sugerem uma elevada frequência de infecção por B. bovis e B. bigemina na população de búfalos estudada. Os testes preliminares do dispositivo de diagnóstico da infecção por B. bovis mostraram alterações na impedância do sistema usado e evidenciaram claramente que o biossensor é capaz de diferencia/detectar os animais infectados, podendo ser explorado para a detecção rápida da B. bovis e estendido também para a detecção de outros parasitas.