Riconosciamo il sapiente aiuto di Annie B. Kristensen, Elisabeth V. Andersen, Belinda B. Bringtoft ed i custodi degli animali presso il Dipartimento di Medicina Sperimentale, Frederiksberg Campus, Facoltà di Scienze della Salute e Scienze Mediche, Università di Copenhagen, Danimarca. Jacob Lønholdt viene ringraziato per l&#39;eccellente assistenza tecnica per le riprese delle procedure. Questo studio è stato sostenuto in parte dal Centro LIFEPHARM di Farmacologia in vivo e Università di Copenhagen.

Le procedure illustrate sono state eseguite secondo protocolli approvati dal danese Animal Sperimentazione Ispettorato sotto il Ministero dell&#39;alimentazione, dell&#39;agricoltura e della pesca. 1 Prelievo di sangue dal laterale safena o tarsale Vena <ol><li> Eseguire questa procedura senza anestesia e con l&#39;aiuto di un investigatore, una gestione della cavia, e una raccolta del campione. </li><li> Radere la regione tarsale della zampa posteriore con un rasoio elettrico fino a quando le vene sono visibili. Assicurarsi che la zona è abbastanza grande da evitare pelliccia interferire con il sito di puntura. </li><li> Posizionare la zampa posteriore in un bagno di acqua tiepida e strofinare delicatamente la gamba, rendendo le vene più dilatati ed evidenti. </li><li> Pulire la gamba con un asciugamano pulito e disinfettare il sito di puntura con una garza o un batuffolo di cotone imbevuto di alcool al 70%. </li><li> Trattenere la cavia ed estendere la gamba posteriore verso il basso, mentre applicando una leggera pressione immediatamente al di sopra del ginocchio to fornire stasi. </li><li> Pungere la vena con un ago 21 G o bisturi. Raccogliere esempio sangue., Utilizzando un tubo di raccolta o capillare. Manipolare delicatamente la gamba per aiutare il flusso di sangue. </li><li> Rilasciare stasi, una volta ottenuto il campione. Applicare un pezzo di garza o cotone lana con una leggera pressione sul sito di puntura finché il sanguinamento si è fermato. </li><li> Ritorna l&#39;animale nella sua gabbia e monitor per 5-10 minuti al fine di garantire l&#39;emostasi. </li></ol> 2 Prelievo di sangue dalla vena dell&#39;orecchio <ol><li> Eseguire questa procedura senza anestesia da solo o preferibilmente con l&#39;aiuto di un collega. </li><li> Disinfettare la superficie dorsale del padiglione auricolare con una garza o un batuffolo di cotone imbevuto di alcool al 70%. </li><li> Applicare una leggera pressione alla base dell&#39;orecchio per fornire stasi. </li><li> Pungere la vena con un ago 21 G o bisturi, e raccogliere il sangue. </li><li> Rilasciare stasi, una volta ottenuto il campione, Applicare un pezzo di garza o cotone idrofilo con gepressione ntle al sito di puntura finché il sanguinamento si è fermato. </li><li> Ritorna l&#39;animale nella sua gabbia e monitor per 5-10 minuti al fine di garantire l&#39;emostasi. </li></ol> 3 Prelievo di sangue dalla giugulare Vena <ol><li> Effettuare questa procedura utilizzando l&#39;aiuto di un investigatore supplementare; una gestione della cavia e una raccolta del campione. </li><li> Anestetizzare la cavia con un anestetico adatto per tutta la durata di questa procedura. Una miscela di tiletamina (0,93 mg / kg), zolazepam (0,93 mg / kg), xilazina (1.49 mg / kg) e butorfanolo (0,06 mg / kg) può essere utilizzato. </li><li> Posizionare la cavia anestetizzata sulla sua schiena e ritrarre delicatamente le zampe anteriori caudalmente. </li><li> Shave e disinfettare (ad es., Con il 70% di alcol), la parte ventrale del collo. </li><li> Rivestire la siringa sia con eparina o EDTA prima del campionamento, al fine di ridurre il rischio di coagulazione. </li><li> Palpare la clavicola e inserire un G 25 (o 23 G) ago montato su una siringa da 1 ml approximately 1 centimetro lateralmente alla linea mediana a livello dell&#39;articolazione della spalla. Tenere la siringa in una leggera inclinazione verso l&#39;alto e mediale, mentre applicando una leggera pressione negativa. Evitare di danneggiare le strutture sottostanti come la vena giugulare si trova abbastanza superficiale. Inclinare il collo della cavia leggermente all&#39;indietro per fornire maggiore stasi della vena. </li><li> Rilasciare le zampe anteriori quando si ottiene il campione di sangue, e alleviare stasi prima che l&#39;ago viene retratto. Usare un pezzo secco di garza o di cotone per applicare una leggera pressione sul sito di puntura. </li><li> Ritorna l&#39;animale in una gabbia di recupero e monitorare al fine di garantire l&#39;emostasi e il recupero dall&#39;anestesia. </li></ol>

<ol>
	<li>DeOgburn, R., 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=Effects+of+increased+dietary+cholesterol+with+carbohydrate+restriction+on+hepatic+lipid+metabolism+in+Guinea+pigs.">Effects of increased dietary cholesterol with carbohydrate restriction on hepatic lipid metabolism in Guinea pigs.</a> Comp.Med. 62 (2), 109-115 (2012).</li><li>Fernandez, M., Volek, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Guinea+pigs+a+suitable+animal+model+to+study+lipoprotein+metabolism,+atherosclerosis+and+inflammation.">Guinea pigs a suitable animal model to study lipoprotein metabolism, atherosclerosis and inflammation.</a> Nutr Metab (Lond). 3 (17), (2006).</li><li>Frikke-Schmidt, H., Tveden-Nyborg, P., Birck, M. M., Lykkesfeldt, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High+dietary+fat+and+cholesterol+exacerbates+chronic+vitamin+C+deficiency+in+guinea+pigs.">High dietary fat and cholesterol exacerbates chronic vitamin C deficiency in guinea pigs.</a> Br J Nutr. 105 (1), 54-61 (2011).</li><li>Hem, A., Smith, A. J., Solberg, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Saphenous+vein+puncture+for+blood+sampling+of+the+mouse,+rat,+hamster,+gerbil,+guinea+pig,+ferret+and+mink.">Saphenous vein puncture for blood sampling of the mouse, rat, hamster, gerbil, guinea pig, ferret and mink.</a> Lab Anim. 32 (4), 364-368 (1998).</li><li>Parasuraman, S., Raveendran, R., Kesavan, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Blood+sample+collection+in+small+laboratory+animals.">Blood sample collection in small laboratory animals.</a> J Pharmacol Pharmacother. 1 (2), 87-93 (2010).</li><li>Guillen, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=FELASA+guidelines+and+recommendations.">FELASA guidelines and recommendations.</a> J Am Assoc Lab Anim Sci. 51 (3), 311-321 (2012).</li><li>Ye, P., Cheah, I. 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. G., Tveden-Nyborg, P., Lykkesfeldt, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prolonged+maternal+vitamin+C+deficiency+overrides+preferential+fetal+ascorbate+transport+but+does+not+influence+perinatal+survival+in+guinea+pigs.">Prolonged maternal vitamin C deficiency overrides preferential fetal ascorbate transport but does not influence perinatal survival in guinea pigs.</a> Br J Nutr. 110 (9), 1573-1579 (2013).</li><li>Greulich, S., 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=Secretory+products+of+guinea+pig+epicardial+fat+induce+insulin+resistance+and+impair+primary+adult+rat+cardiomyocyte+function.">Secretory products of guinea pig epicardial fat induce insulin resistance and impair primary adult rat cardiomyocyte function.</a> J Cell Mol Med. 15 (11), 2399-2410 (2011).</li><li>Swifka, J., Weiss, J., Addicks, K., Eckel, J., Rosen, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Epicardial+fat+from+guinea+pig+a+model+to+study+the+paracrine+network+of+interactions+between+epicardial+fat+and+myocardium.">Epicardial fat from guinea pig a model to study the paracrine network of interactions between epicardial fat and myocardium.</a> Cardiovasc Drugs Ther. 22 (2), 107-114 (2008).</li><li>Pilny, A. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clinical+hematology+of+rodent+species.">Clinical hematology of rodent species.</a> Vet.Clin North Am Exot Anim Pract. 11 (3), 523-vii (2008).</li><li>Franco, N. H., Olsson, I. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Scientists+and+the+3Rs:+attitudes+to+animal+use+in+biomedical+research+and+the+effect+of+mandatory+training+in+laboratory+animal+science.">Scientists and the 3Rs: attitudes to animal use in biomedical research and the effect of mandatory training in laboratory animal science.</a> Lab Anim. 48 (1), 50-60 (2014).</li><li>Van, L. J., 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=eAssessing+the+application+of+the+3Rs+a+survey+among+animal+welfare+officers+in+The+Netherlands.">eAssessing the application of the 3Rs a survey among animal welfare officers in The Netherlands.</a> Lab Anim. 47 (3), 210-219 (2013).</li><li>Sloan, R. C., 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=High+doses+of+ketamine-xylazine+anesthesia+reduce+cardiac+ischemia-reperfusion+injury+in+guinea+pigs.">High doses of ketamine-xylazine anesthesia reduce cardiac ischemia-reperfusion injury in guinea pigs.</a> J Am Assoc Lab Anim Sci. 50 (3), 349-354 (2011).</li><li>Jacobson, C. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+anaesthetic+regimen+for+surgical+procedures+in+guinea+pigs.">A novel anaesthetic regimen for surgical procedures in guinea pigs.</a> Lab Anim. 35 (3), 271-276 (2001).</li><li>Saha, J. K., Xia, J., Grondin, J. M., Engle, S. K., Jakubowski, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acute+hyperglycemia+induced+by+ketamine+xylazine+anesthesia+in+rats+mechanisms+and+implications+for+preclinical+models.">Acute hyperglycemia induced by ketamine xylazine anesthesia in rats mechanisms and implications for preclinical models.</a> Exp Biol Med(Maywood). 230 (10), 777-784 (2005).</li></ol>

Gli autori non hanno nulla da rivelare. Tutte le procedure sono state approvate dalla legge Sperimentazione Animale della Danimarca, che è in accordo con il Consiglio d&#39;Europa Convenzione ETS 123.

Rispetto ai topi e ratti, cavie sono timorosi per natura e devono essere affrontati con calma e gestite correttamente, al fine di ridurre lo stress. Da un punto di vista del benessere degli animali, agendo con cura è naturalmente una priorità; tuttavia, questo sarà anche ridurre il rischio di effetti di ansia associati sui dati raccolti ad es., aumento dei livelli ematici di corticosteroidi e di glucosio 12. Allo stesso modo, la gestione quotidiana sarà abituare gli animali a procedure normali e ...

La cavia è un modello valido e convalidato animale sperimentale a causa di un numero di somiglianze biologiche per l&#39;uomo, come l&#39;esigenza di un approvvigionamento alimentare di vitamina C, lipoproteine ​​plasmatiche paragonabili enzimi che metabolizzano e profili delle lipoproteine, così come similitudini in comune con placentazione umana e sviluppo prenatale 1-3,7,8. Questo rende la cavia un modello attraente e adatto per studiare gli effetti delle malattie dieta associata come la malattia non alcolica steatosi epatica, malattie cardiovascolari e gli effetti putativi di carenza di vitamina C, ma anche l&#39;effetto di esempio., Intervento dietetico materno sulla via di sviluppo prole. Ripetuto prelievo di sangue è spesso richiesto sia lungo e breve termine in studi in vivo. A causa delle differenze anatomiche specie-specifici è necessaria la modulazione della tecnica di campionamento al fine di ottenere campioni di sangue di cavie rispetto ad altri modelli di roditori <sup&gt; 4,5. Questo include la variazione nel corso di vene e arterie, e anche che le cavie non hanno una coda, quindi hanno i requisiti e le diverse opzioni sia per il campionamento e la manipolazione. Campionamento dalla safena o vena tarso, induce il minimo disagio e lo stress negli animali e non richiede anestesia 3,9. Questa tecnica permette ripetuta campioni di piccole quantità di sangue (100-400 ml), ad es., Per determinare i marcatori biochimici nel plasma. Se un campione ancora più piccolo è sufficiente (50-100 ml), questo può facilmente essere ottenuto pungere la vena dell&#39;orecchio. Questa tecnica può anche essere effettuata in animali coscienti ed è utile ad esempio quando si misura la glicemia 10,11. Campionamento dalla vena giugulare consente la raccolta di grandi quantità di sangue (1-2 ml), ma richiede l&#39;anestesia. Qui, vi presentiamo quattro diverse tecniche di campionamento del sangue sia per cavie coscienti o anestetizzati. Il procedures sono tutte le procedure non-terminali a condizione che i volumi di campione e il numero di campioni non superino le linee guida per la raccolta del sangue negli animali da laboratorio 6. E &#39;nell&#39;interesse della buona scienza, nonché di benessere degli animali che lo stress dovrebbe essere ridotto al minimo. Di conseguenza, solo ben addestrato e personale competente deve eseguire le procedure sugli animali.

<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

Le cavie possiedono diverse analogie biologiche per l&#39;uomo e sono convalidati modelli animali sperimentali 1-3. Tuttavia, l&#39;uso di cavie rappresenta attualmente una zona relativamente ristretta dei dati di ricerca e descrittivi sulla metodologia specifica è corrispondentemente scarsa. Le caratteristiche anatomiche di cavie sono leggermente diversi da altri modelli di roditori, quindi la modulazione di tecniche di campionamento per ospitare per specie-specifiche differenze, ad es., Rispetto ai topi e ratti, sono necessari per ottenere campioni di qualità sufficiente e di alta. Per quanto sia lungo e breve termine in studi in vivo spesso richiedono ripetuto sangue campionamento la scelta della tecnica deve essere ben considerato, al fine di ridurre lo stress e il disagio negli animali, ma anche per garantire la sopravvivenza, nonché la conformità con i requisiti di dimensione del campione e accessibilità. Campioni di sangue venoso possono essere ottenuti ad un certo numero di siti in cavie ad es., La safena e giugularevene, ogni tecnica contenente sia vantaggi che svantaggi 4,5. Qui, vi presentiamo quattro diverse tecniche di campionamento del sangue sia per cavie coscienti o anestetizzati. Le procedure sono tutte le procedure non-terminali a condizione che i volumi di campione e il numero di campioni non superino le linee guida per la raccolta del sangue negli animali da laboratorio 6. Tutti i metodi descritti sono stati accuratamente testati e applicati per ripetute in vivo prelievo di sangue in studi all&#39;interno della nostra struttura di ricerca.

Quattro diversi approcci alla non-terminali vivo tecniche di campionamento del sangue in entrambi in cavie coscienti o anestetizzati sono stati presentati. Figura 1 illustra il corso della safena laterale e vena tarsale nella cavia e possibili siti di puntura. Vene dell&#39;orecchio visibili sulla superficie dorsale di un orecchio cavia sono illustrati nella Figura 2 Queste vene possono essere usate per il prelievo di sangue quando è richiesto solo un volume molto pic...

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

Anche se un modello noto, la cavia rappresenta attualmente una nicchia in scienze animali sperimentali e dati limitati sono disponibili sul esecuzione della maggior parte delle procedure. Qui vi presentiamo quattro diversi approcci alla non-terminale in tecniche di campionamento del sangue in vivo su cavie sia coscienti o anestetizzati.

Non terminale Tecniche di prelievo di sangue in Guinea Pigs

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 Views.  University of Copenhagen.  Anche se un modello noto, la cavia rappresenta attualmente una nicchia in scienze animali sperimentali e dati limitati sono disponibili sul esecuzione della maggior parte delle procedure. Qui vi presentiamo quattro diversi approcci alla non-terminale in tecniche di campionamento del sangue in vivo su cavie sia coscienti o anestetizzati. 

Video: 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.

Disegno di sangue di ratti attraverso la vena safena e da puntura cardiaca

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: ...

Ricerca

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.

Ottimizzazione del interferone-gamma ELISpot analisi di risposte a cellula T misura nel modello cavia dopo la vaccinazione

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, ...

Immunologia e infezioni

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.

Puntura della vena succlavia come metodo alternativo di raccolta del campione di sangue in ratti

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.

Un approccio chirurgico rivisto per indurre idramici endolymphatic in 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) ...

Neuroscienze

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.

Il campionamento continuo del sangue nella tomografia a emissione di positroni di piccole dimensioni/tomografia computerizzata consente la misurazione della funzione di input arteriosa

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.

Prelievo di sangue dalla vena della coda laterale del Topo

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.

Campionamento ripetitivo di sangue dalla vena succlavia del ratto cosciente

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.

Misurazione transdermica della velocità di filtrazione glomerulare nei suinetti ventilati meccanicamente

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

Studio comparativo delle tecniche di raccolta del sangue nei ratti

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.

Autore in primo piano: Metodi di raccolta del sangue dipendenti dal contesto nei ratti 

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.

Puntura modificata della vena della coda e della vena del pene per il prelievo di sangue nel modello di ratto

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

Non terminale Tecniche di prelievo di sangue in Guinea Pigs

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Il campionamento continuo del sangue nella tomografia a emissione di positroni di piccole dimensioni/tomografia computerizzata consente la misurazione della funzione di input arteriosa

Prelievo di sangue dalla vena della coda laterale del Topo

Campionamento ripetitivo di sangue dalla vena succlavia del ratto cosciente

Misurazione transdermica della velocità di filtrazione glomerulare nei suinetti ventilati meccanicamente

Puntura modificata della vena della coda e della vena del pene per il prelievo di sangue nel modello di ratto

Puntura modificata della vena della coda e della vena del pene per il prelievo di sangue nel modello di ratto