Name: non-terminal blood sampling techniques in guinea pigs
Uploaded: 2014-10-11T14:00:00.000+00:00
Duration: 7 min 58 s
Description: Watch this Scientific Journal Video about Non-Terminal Blood Sampling Techniques in Guinea Pigs at JoVE.com

Reconhecemos a assistência hábil de Annie B. Kristensen, Elisabeth V. Andersen, Belinda B. Bringtoft e os cuidadores de animais no Departamento de Medicina Experimental, Frederiksberg Campus da Faculdade de Saúde e Ciências Médicas da Universidade de Copenhague, na Dinamarca. Jacob Lønholdt está agradeceu por sua assistência técnica excelente em filmar os procedimentos. Este estudo foi financiado em parte pelo Centro LIFEPHARM de farmacologia in vivo e da Universidade de Copenhagen.

Os procedimentos mostrados foram realizados de acordo com protocolos aprovados pelo animal dinamarquês Experimentação Inspecção no âmbito do Ministério da Alimentação, Agricultura e Pescas. 1. Sangue Amostragem do lateral safena ou Tarsal Veia <ol><li> Execute este procedimento sem anestesia e com a ajuda de um investigador, uma manipulação a cobaia, e uma coleta de amostra. </li><li> Raspar a região tarsal da perna com um barbeador elétrico até que as veias são visíveis. Certifique-se de que a área é grande o suficiente para evitar interferir com a pele no local de punção. </li><li> Coloque a perna em um banho de água morna e esfregue suavemente a perna, fazendo com que as veias mais dilatada e evidente. </li><li> Limpe a perna com uma toalha limpa e desinfectar o local da punção com gaze ou um chumaço de algodão umedecido com álcool 70%. </li><li> Contenha a cobaia e estender a perna para baixo, aplicando pressão suave imediatamente acima do joelho to fornecer estase. </li><li> Por punção da veia com uma agulha de 21 G ou lanceta. Recolha de sangue, por exemplo., Usando um tubo de recolha ou capilar. Gentilmente manipular a perna para ajudar o fluxo de sangue. </li><li> Solte estase, uma vez que a amostra foi obtida. Aplique um pedaço de gaze ou algodão, com uma leve pressão ao local da punção até que o sangramento parou. </li><li> Retorne o animal para sua gaiola e monitor por 5-10 min para assegurar a hemostasia. </li></ol> 2. coleta de sangue da veia da orelha <ol><li> Executar este procedimento, sem anestesia, quer isoladamente ou, de preferência com a ajuda de um colega. </li><li> Desinfetar a superfície dorsal da orelha com gaze ou um chumaço de algodão umedecido com álcool 70%. </li><li> Aplicar uma leve pressão para a base da orelha para proporcionar estase. </li><li> Por punção da veia com uma agulha de 21 G ou lanceta, e coleta de sangue. </li><li> Solte estase, uma vez que a amostra foi obtida, aplique um pedaço de gaze ou algodão com gepressão ntle ao local da punção até que o sangramento parou. </li><li> Retorne o animal para sua gaiola e monitor por 5-10 min para assegurar a hemostasia. </li></ol> 3. Sangue Amostragem da veia jugular <ol><li> Realizar este procedimento com a ajuda de um investigador adicional; uma manipulação da cobaia e uma coleta de amostra. </li><li> Anestesiar a cobaia com um anestésico adequado durante toda a duração deste procedimento. Uma mistura de tiletamina (0,93 mg / kg), zolazepam (0,93 mg / kg), xilazina (1,49 mg / kg) e butorfanol (0,06 mg / kg) pode ser usado. </li><li> Coloque a cobaia anestesiada em suas costas e retirar delicadamente as pernas dianteiras caudal. </li><li> Raspar e desinfectar (por ex., Com 70% de álcool) a parte ventral do pescoço. </li><li> Cubra a seringa com heparina ou EDTA antes da amostragem, a fim de reduzir o risco de coagulação. </li><li> Palpar a clavícula e inserir um 25 G (ou 23 G) agulha montada em um 1 ml ap seringaximadamente 1 cm lateral à linha média ao nível da articulação do ombro. Manter a seringa em um pequeno ângulo para cima e medial enquanto aplica uma pressão ligeiramente negativa. Evite danificar as estruturas subjacentes como a veia jugular está situado muito superficial. Inclinação do gargalo da cobaia ligeiramente para trás, para proporcionar o aumento da estase venosa. </li><li> Libertar as pernas da frente, quando a amostra de sangue é obtida, e aliviar a estase antes de a agulha é retraída. Use um pedaço de gaze seca ou algodão para aplicar pressão suave para o local da punção. </li><li> Retorne o animal para uma gaiola de recuperação e monitoramento para garantir a hemostasia e recuperação da anestesia. </li></ol>

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K., Halliwell, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High+fat+diets+and+pathology+in+the+guinea+pig+Atherosclerosis+or+liver+damage.">High fat diets and pathology in the guinea pig Atherosclerosis or liver damage.</a> Biochim Biophys Acta. 1832 (2), 355-364 (2013).</li><li>Tveden-Nyborg, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Maternal+vitamin+C+deficiency+during+pregnancy+persistently+impairs+hippocampal+neurogenesis+in+offspring+of+guinea+pigs.">Maternal vitamin C deficiency during pregnancy persistently impairs hippocampal neurogenesis in offspring of guinea pigs.</a> PLoS One. 7 (10), 12-18391 (2012).</li><li>Schjoldager, J. 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Os autores não têm nada a revelar. Todos os procedimentos foram aprovados pela Lei de Experimentação Animal da Dinamarca, que está de acordo com o Conselho da Europa Convenção ETS 123.

Em comparação com camundongos e ratos, cobaias estão com medo, por natureza e deve ser abordada com calma e tratada corretamente, a fim de reduzir o estresse. Do ponto de vista do bem-estar animal, agindo com cuidado é naturalmente uma prioridade; no entanto, este também irá reduzir o risco de efeitos associados a ansiedade sobre o por exemplo, dados coletados., aumento dos níveis sanguíneos de corticosteróides e glicose 12. Da mesma forma, a movimentação diária irá habituar os animais c...

A cobaia um modelo experimental animal valioso e validado devido a um número de semelhanças biológicos para os seres humanos, tais como a exigência de uma fonte dietética de vitamina C, a lipoproteína plasmática comparável enzimas que metabolizam e perfis de lipoproteína, bem como semelhanças compartilhadas com placentação humana e desenvolvimento pré-natal 1-3,7,8. Isso faz com que a cobaia um modelo atraente e adequado para estudar os efeitos de doenças associadas à dieta, como a doença não alcoólica do fígado gorduroso, doenças cardiovasculares e efeitos putativos de deficiência de vitamina C, mas também o efeito de, por exemplo., A intervenção dietética materna sobre o desenvolvimento prole. Coleta de sangue repetida muitas vezes é necessária tanto em curto e longo prazo em estudos in vivo. Devido às diferenças anatómicas específicas de espécies é necessário modulação da técnica de amostragem, a fim de obter amostras de sangue em cobaias, em comparação com outros modelos de roedores <sup&gt; 4,5. Isto inclui a variação ao longo de veias e artérias, e também que as cobaias não tem uma cauda, ​​assim, ter diferentes requisitos e opções de amostragem e de manuseamento. Amostragem do safena ou veia do tarso, induz o mínimo de desconforto e estresse no animal e não requer anestesia 3,9. Esta técnica permite que as amostras de pequenas quantidades de sangue (100-400 ul), por exemplo, repetido., Para determinar os marcadores bioquímicos no plasma. Se um tamanho ainda mais pequeno da amostra é suficiente (50-100 ul), este pode facilmente ser obtido por punção da veia da orelha. Esta técnica também pode ser levada a cabo em animais conscientes e é útil, por exemplo, quando a medição da glucose no sangue 10,11. A amostragem a partir da veia jugular permite a recolha de maiores quantidades de sangue (1-2 ml), mas requer anestesia. Aqui, apresentamos quatro técnicas de amostragem de sangue diferentes para ambos cobaias conscientes ou anestesiados. O procedures são todos os procedimentos não-terminal na condição de os volumes de amostra e número de amostras não exceder as diretrizes para coleta de sangue em animais de laboratório 6. É no interesse da boa ciência, bem como de bem-estar animal que o estresse deve ser reduzido ao mínimo. Consequentemente, apenas bem treinados e pessoal competente deve executar os procedimentos com animais.

<table><tbody><tr><td>Electric shaver</td><td>Wella (Contura), Denmark</td><td>273207</td><td>Alternative product may be used. Multiple suppliers</td></tr><tr><td>Syringes&amp;nbsp;</td><td>B.Braun-Omnifix-F, Germany</td><td>9161406V</td><td>Alternative product may be used. Multiple suppliers</td></tr><tr><td>Needles</td><td>BD Microlance, Spain</td><td>301156</td><td>Alternative product may be used. Multiple suppliers</td></tr><tr><td>Microvette collection tube&amp;nbsp;</td><td>Sarstedt, D-51588N&amp;uuml;mbrecht, Germany</td><td>201,288,100</td><td>Alternative product may be used. Multiple suppliers</td></tr><tr><td>Accu-Check analyzer</td><td>Roche A/S Diagnostics, Denmark</td><td></td><td>Alternative product may be used. Multiple suppliers</td></tr><tr><td>Heparin 5000 IE/ml</td><td>LEO Pharma, Denmark</td><td></td><td>Alternative product may be used. Multiple suppliers</td></tr><tr><td>Anesthesia</td><td>Zoletil 50, Chemvet, Denmark</td><td></td><td>Alternative product may be used. Multiple suppliers</td></tr><tr><td>Anesthesia</td><td>Narcoxyl vet 20 mg/ml, Intervet, Denmark</td><td></td><td>Alternative product may be used. Multiple suppliers</td></tr><tr><td>Anesthesia</td><td>Torbugesic vet 10 mg/ml, Scanvet, Denmark</td><td></td><td>Alternative product may be used. Multiple suppliers</td></tr><tr><td>Neutral eye ointment</td><td></td><td></td><td>Alternative product may be used. Multiple suppliers</td></tr><tr><td>Gauze or a cotton swab&amp;nbsp;</td><td></td><td></td><td>Alternative product may be used. Multiple suppliers</td></tr><tr><td>Disinfectant (70% alcohol)</td><td></td><td></td><td>Alternative product may be used. Multiple suppliers</td></tr></tbody></table>

non-terminal blood sampling techniques in guinea pigs

As cobaias possuem várias semelhanças biológicas para os seres humanos e são validados modelos animais experimentais 1-3. No entanto, o uso de cobaias representa atualmente uma área relativamente estreita de dados de pesquisa e descritivos sobre metodologia específica é correspondentemente escasso. As características anatômicas de cobaias são um pouco diferentes de outros modelos de roedores, daí modulação de técnicas de amostragem para acomodar as diferenças espécie-específicos, por exemplo., Em comparação com camundongos e ratos, são necessários para obter amostras suficientes e de alta qualidade. Como tanto a longo e curto prazo em estudos in vivo, muitas vezes requerem repetidas sangue amostragem a escolha da técnica deve ser bem considerado, a fim de reduzir o estresse e desconforto nos animais, mas também para garantir a sobrevivência, bem como o cumprimento dos requisitos de tamanho da amostra e acessibilidade. Amostras de sangue venoso pode ser obtido em uma série de sites em cobaias por exemplo., A safena e jugularveias, cada uma das técnicas que contém tanto vantagens como desvantagens 4,5. Aqui, apresentamos quatro técnicas de amostragem de sangue diferentes para ambos cobaias conscientes ou anestesiados. Os procedimentos são todos os procedimentos não-terminal na condição de os volumes de amostra e número de amostras não exceder as diretrizes para coleta de sangue em animais de laboratório 6. Todos os métodos descritos têm sido exaustivamente testado e aplicado para repetido em amostras de sangue vivo em estudos dentro do nosso centro de pesquisa.

Quatro abordagens diferentes para in vivo de técnicas de amostragem de sangue não-terminal, nem em cobaias conscientes ou anestesiados foram apresentados. Figura 1 ilustra o curso da safena lateral e veia tarsal na cobaia e possíveis locais de punção. Veias da orelha visíveis na superfície dorsal de uma orelha da cobaia são ilustrados na Figura 2 Estas veias pode ser utilizado para a amostragem de sangue, quando apenas um volume muito pequeno, é necessária. A posiçã...

Watch this Scientific Journal Video about Non-Terminal Blood Sampling Techniques in Guinea Pigs at JoVE.com

Non-Terminal Blood Sampling Techniques in Guinea Pigs

Apesar de ser um modelo conhecido, a cobaia representa atualmente um nicho em ciências experimentais em animais e dados limitada disponível sobre a execução da maioria dos procedimentos. Apresentamos aqui quatro abordagens diferentes para não-terminal em técnicas de amostragem de sangue vivo em qualquer cobaias conscientes ou anestesiados.

Técnicas de coleta de sangue não terminal em cobaias

Guinea pigs possess several biological similarities to humans and are validated experimental animal models1-3. However, the use of guinea pigs currently ...

non-terminal-blood-sampling-techniques-in-guinea-pigs

Universidad de Cartagena UDEC

Research

JoVE Journal

Biology

37.3K visualizações.  University of Copenhagen.  Apesar de ser um modelo conhecido, a cobaia representa atualmente um nicho em ciências experimentais em animais e dados limitada disponível sobre a execução da maioria dos procedimentos. Apresentamos aqui quatro abordagens diferentes para não-terminal em técnicas de amostragem de sangue vivo em qualquer cobaias conscientes ou anestesiados. 

Vídeo: Non-Terminal Blood Sampling Techniques in Guinea Pigs

Drawing Blood from Rats through the Saphenous Vein and by Cardiac Puncture

Drawing blood from rodents is necessary for a large number of both in vitro and in vivo studies. Sites of blood draws are numerous in rodents: retro-orbital sinus, jugular vein, maxillary vein, saphenous vein, heart. Each technique has its advantages and disadvantages, and some are not approved any more in some countries (e.g., retro-orbital draws in Holland). A discussion of different techniques for drawing blood are available 1-3.
Here, we present two techniques for drawing blood from rats, each with its specific applications.
Blood draw from the saphenous vein, provided it is done properly, induces minimal distress in animals and does not require anesthesia. This technique allows repeated draws of small amounts of blood, such as needed for pharmacokinetic studies 4,5, determining plasma chemistry, or blood counts 6.
Cardiac puncture allows the collection of large amounts of blood from a single animal (up to 10 ml of blood can be drawn from a 150 g rat). This technique is therefore very useful as a terminal procedure when drawing blood from the saphenous would not provide a large enough sample. We use cardiac puncture when we need sufficient amounts of serum from a specific strain of rats to grow T lymphocyte lines in vitro 4-9.

Blood draws are necessary in a large number of studies, for example to study the pharmacokinetics profile of a compound. Here, we demonstrate how to draw blood from rats using two techniques: blood draw from the saphenous vein or by cardiac puncture.

Desenho de sangue de ratos através da veia safena e por punção cardíaca

Drawing blood from rodents is necessary for a large number of both in vitro and in vivo studies. Sites of blood draws are numerous in rodents: ...

Biologia

Optimized Interferon-gamma ELISpot Assay to Measure T Cell Responses in the Guinea Pig Model after Vaccination

The guinea pig has played a pivotal role as a relevant small animal model in the development of vaccines for infectious diseases such as tuberculosis, influenza, diphtheria, and viral hemorrhagic fevers. We have demonstrated that plasmid-DNA (pDNA) vaccine delivery into the skin elicits robust humoral responses in the guinea pig. However, the use of this animal to model immune responses was somewhat limited in the past due to the lack of available reagents and protocols to study T cell responses. T cells play a pivotal role in both immunoprophylactic and immunotherapeutic mechanisms. Understanding T cell responses is crucial for the development of infectious disease and oncology vaccines and accommodating delivery devices. Here we describe an interferon-gamma (IFN-&#947;) enzyme-linked immunospot (ELISpot) assay for guinea pig peripheral blood mononuclear cells (PBMCs). The assay enables researchers to characterize vaccine-specific T-cell responses in this important rodent model. The ability to assay cells isolated from the peripheral blood provides the opportunity to track immunogenicity in individual animals.

The development of the interferon-gamma ELISpot assay for guinea pig PBMCs allows the characterization of T-cell responses in this highly relevant model for studying infectious diseases. We have applied the assay to measure T cell responses associated with intradermal delivery of DNA vaccines.

Ensaio de ELISpot otimizado de Interferon-gama às respostas de célula T medida no modelo de porquinho da Índia após a vacinação

The guinea pig has played a pivotal role as a relevant small animal model in the development of vaccines for infectious diseases such as tuberculosis, ...

Imunologia e Infecção

Subclavian Vein Puncture As an Alternative Method of Blood Sample Collection in Rats

Blood collection with enough blood volume is essential in animal experiments. Blood collection from the tail vein of rats is popular and less stressful compared to other more aggressive methods such as retro-orbital plexus sample collection. However, this blood collection method is sometimes limited by an unsatisfactory success rate. Here, we introduce a method for blood collection through the subclavian vein puncture. The subclavian vein is located just under the clavicle and this vein is large enough to fulfill the volume requirements of blood collection. Our results show that this method is safe and applicable for blood collection sampling with the required blood volume. Blood collection through the subclavian vein puncture could serve as an alternative blood collection method in case of failed tail vein blood sampling in rats.

Here, we present a protocol to collect blood samples from the rat subclavian vein.

Punção da veia subclávia como um método alternativo de coleta de amostra de sangue em ratos

Blood collection with enough blood volume is essential in animal experiments. Blood collection from the tail vein of rats is popular and less stressful ...

Medicina

A Revised Surgical Approach to Induce Endolymphatic Hydrops in the Guinea Pig

Endolymphatic hydrops is an enlargement of scala media that is most often associated with Meniere&#39;s disease, though the pathophysiologic mechanism(s) remain unclear. In order to adequately study the attributes of endolymphatic hydrops, such as the origins of low-frequency hearing loss, a reliable model is needed. The guinea pig is a&#160;good model because it hears in the low-frequency regions that are putatively affected by endolymphatic hydrops. Previous research has demonstrated that endolymphatic hydrops can be induced surgically via intradural or extradural approaches that involve drilling on the endolymphatic duct and sac. However, whether it was possible to create an endolymphatic hydrops model using an extradural approach that avoided dangerous drilling on the endolymphatic duct and sac was unknown. The objective of this study was to demonstrate a revised extradural approach to induce experimental endolymphatic hydrops at 30 days post-operatively by obliterating the endolymphatic sac and injuring the endolymphatic duct with a fine pick. The sample size consisted of seven guinea pigs. Functional measurements of hearing were made and temporal bones were subsequently harvested for histologic analysis. The approach had a success rate of 86% in achieving endolymphatic hydrops. The risk of cerebrospinal fluid leak was minimal. No perioperative deaths or injuries to the posterior semicircular canal occurred in the sample. The presented method demonstrates a safe and reliable way to induce endolymphatic hydrops at a relatively quick time point of 30 days. The clinical implications are that the presented method provides a reliable model to further explore the origins of low-frequency hearing loss that can be associated endolymphatic hydrops.

This article demonstrates an extradural approach to obliterate the guinea pig endolymphatic sac and injure the endolymphatic duct with a fine pick in order to induce experimental endolymphatic hydrops.

Uma abordagem cirúrgica revisada para induzir hidrops endoglicêsticos na Cobaia

Endolymphatic hydrops is an enlargement of scala media that is most often associated with Meniere&#39;s disease, though the pathophysiologic mechanism(s) ...

Neurociência

Continuous Blood Sampling in Small Animal Positron Emission Tomography/Computed Tomography Enables the Measurement of the Arterial Input Function

For quantitative analysis and bio-kinetic modeling of positron emission tomography/computed tomography (PET/CT) data, the determination of the temporal blood time-activity concentration also known as arterial input function (AIF) is a key point, especially for the characterization of animal disease models and the introduction of newly developed radiotracers. The knowledge of radiotracer availability in the blood helps to interpret PET/CT-derived data of tissue activity. For this purpose, online blood sampling during the PET/CT imaging is advisable to measure the AIF. In contrast to manual blood sampling and image-derived approaches, continuous online blood sampling has several advantages. Besides the minimized blood loss, there is an improved resolution and a superior accuracy for the blood activity measurement. However, the major drawback of online blood sampling is the costly and time-consuming preparation to catheterize the femoral vessels of the animal. Here, we describe an easy and complete workflow for catheterization and continuous blood sampling during small animal PET/CT imaging and compared it to manual blood sampling and an image-derived approach. Using this highly-standardized workflow, the determination of the fluorodeoxyglucose ([18F]FDG) AIF is demonstrated. Further, this procedure can be applied to any radiotracer in combination with different animal models to create fundamental knowledge of tracer kinetic and model characteristics. This allows a more precise evaluation of the behavior of pharmaceuticals, both for diagnostic and therapeutic approaches in the preclinical research of oncological, neurodegenerative and myocardial diseases.

Here a protocol for continuous blood sampling during PET/CT imaging of rats to measure the arterial input function (AIF) is described. The catheterization, the calibration and setup of the system and the data analysis of the blood radioactivity are demonstrated. The generated data provide input parameters for subsequent bio-kinetic modeling.

A amostragem contínua do sangue no tomography de emissão de Positron animal pequeno/tomography computado permite a medida da função de entrada arterial

For quantitative analysis and bio-kinetic modeling of positron emission tomography/computed tomography (PET/CT) data, the determination of the temporal ...

Sampling Blood from the Lateral Tail Vein of the Rat

Blood samples are commonly obtained in many experimental contexts to measure targets of interest, including hormones, immune factors, growth factors, proteins, and glucose, yet the composition of the blood is dynamically regulated and easily perturbed. One factor that can change the blood composition is the stress response triggered by the sampling procedure, which can contribute to variability in the measures of interest. Here we describe a procedure for blood sampling from the lateral tail vein in the rat. This procedure offers significant advantages over other more commonly used techniques. It permits rapid sampling with minimal pain or invasiveness, without anesthesia or analgesia. Additionally, it can be used to obtain large volume samples (upwards of 1 ml in some rats), and it may be used repeatedly across experimental days. By minimizing the stress response and pain resulting from blood sampling, measures can more accurately reflect the true basal state of the animal, with minimal influence from the sampling procedure itself.

Blood samples are useful for assessing biomarkers of physiological states or disease in vivo. Here we describe the methodology to sample blood from the lateral tail vein in the rat. This method provides rapid samples with minimal pain and invasiveness.

A amostragem de sangue a partir da veia lateral da cauda do rato

Blood samples are commonly obtained in many experimental contexts to measure targets of interest, including hormones, immune factors, growth factors, ...

Repetitive Blood Sampling from the Subclavian Vein of Conscious Rat

Rats are widely used in pharmacokinetics (PK) and toxicokinetic (TK) studies that need to collect a certain amount of blood at specific time points to detect drug exposure. The rat blood sampling method determines the quality of the plasma and further affects the precision of the test results. The subclavian vein blood collection method described in this protocol collects blood samples repeatedly in the conscious state of animals to meet the needs of PK and TK tests. The skills of restraint handling and appropriate procedure of needle incision ensure the success rate of blood collection. It is easy to operate while ensuring the quality of plasma and at the same time catering to animal welfare. However, this method requires skilled operation, and an improper one may cause animal weakness, pain, lameness, and even mortality. The current method has been used in the testing facility for a 4-week oral toxicity study in Sprague Dawley (SD) rats with TK. The maximum amount of blood collected within 24 h did not exceed 20% of the animal's total blood. The animals' body weight was more than 200 g for males and females. The data showed that the animals' bodyweight increased steadily every week, and the clinical observation was normal after repetitive sample collection.

The present protocol describes a simple and efficient method for collecting blood from the subclavian vein in rats. It enables rapid, timely, and easily identifiable sampling without anesthesia and obtains high-quality blood through repetitive sample collection.

Amostragem Repetitiva de Sangue da Veia Subclávia de Rato Consciente

Rats are widely used in pharmacokinetics (PK) and toxicokinetic (TK) studies that need to collect a certain amount of blood at specific time points to ...

Transdermal Measurement of Glomerular Filtration Rate in Mechanically Ventilated Piglets

Transdermal measurement of glomerular filtration rate (GFR) has been used to evaluate kidney function in conscious animals. This technique is well established in rodents to study acute kidney injury and chronic kidney disease. However, GFR measurement using the transdermal system has not been validated in pigs, a species with a similar renal system to humans. Hence, we investigated the effect of sepsis on transdermal GFR in anesthetized and mechanically ventilated neonatal pigs. Polymicrobial sepsis was induced by cecal ligation and puncture (CLP). The transdermal GFR measurement system consisting of a miniaturized fluorescence sensor was attached to the pig's shaved skin to determine the clearance of fluorescein-isothiocyanate (FITC) conjugated sinistrin, an intravenously injected GFR tracer. Our results show that at 12 h post-CLP, serum creatinine increased with a decrease in GFR. This study demonstrates, for the first time, the utility of the transdermal GFR approach in determining renal function in mechanically ventilated, neonatal pigs.

Glomerular filtration rate (GFR) is the ideal marker for assessing kidney function. However, the standard measurement method using inulin injection with serial blood and urine analysis is impractical. This article delineates a practical method to measure GFR transdermally in piglets.

Medição Transdérmica da Taxa de Filtração Glomerular em Leitões Ventilados Mecanicamente

Transdermal measurement of glomerular filtration rate (GFR) has been used to evaluate kidney function in conscious animals. This technique is well ...

Estudo Comparativo de Técnicas de Coleta de Sangue em Ratos

Modified Tail Vein and Penile Vein Puncture for Blood Sampling in the Rat Model

Blood samples are required in most experimental animal designs to assess various hematological parameters. This paper presents two procedures for blood collection in rats: the lateral tail vein puncture and the dorsal penile vein puncture, which offer significant advantages over other previously described techniques. This study shows that these two procedures allow for fast sampling (under 10 min) and yield sufficient blood volumes for most assays (202 &#956;L &#177; 67.7 &#956;L). The dorsal penile vein puncture must be done under anesthesia, whereas the lateral tail vein puncture can be done on a conscious, restrained animal.
Alternating these two techniques, therefore, enables blood draw in any situation. While it is always recommended for an operator to be assisted during a procedure to ensure animal welfare, these techniques require only a single operator, unlike most blood sampling methods that require two. Moreover, whereas these previously described methods (e.g., jugular stick, subclavian vein blood draw) require extensive prior training to avoid harm to or death of the animal, tail vein and dorsal penile vein puncture are rarely fatal. For all these reasons, and according to the context (e.g., for studies including male rats, during the perioperative or immediate postoperative period, for animals with thin tail veins), both techniques can be used alternately to enable repeated blood draws.

Autor em destaque: métodos de coleta de sangue dependentes do contexto em ratos 

Here, we present a protocol to offer rapid, easy, and reliable blood collection alternatives for the rat model. We describe three different blood sampling methods according to the context: tail vein puncture under anesthesia or on a conscious animal, and dorsal penile vein puncture under anesthesia.

Veia Caudal e Punção Peniana Modificadas para Coleta de Sangue no Modelo de Rato

Blood samples are required in most experimental animal designs to assess various hematological parameters. This paper presents two procedures for blood ...

Técnicas de coleta de sangue não terminal em cobaias

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