<ol>
	<li>Baker, D. G., Fox, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chapter+23.">Chapter 23.</a> The Mouse in Biomedical Research: Diseases. Diseases Vol. 2, (2007).</li><li>Baker, D. G., Baker, D. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chapter+11.">Chapter 11.</a> Flynn's Parasites of Laboratory Animals. , 303-398 (2007).</li><li>Owen, D. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Parasites of Laboratory Animals. 12, (1992).</li><li>Pritchett, K. R., Fox, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chapter+22.">Chapter 22.</a> The Mouse in Biomedical Research: Diseases. Diseases Vol. 2, (2007).</li><li>Smith, P. H., Wiles, S. E., Malone, J. B., Monahan, C. M., Baker, D. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chapter+1.">Chapter 1.</a> Flynn's Parasites of Laboratory Animals. , 1-13 (2007).</li><li>Wasson, K., Fox, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chapter+21.">Chapter 21.</a> The Mouse in Biomedical Research: Diseases. Vol. 2, 517-550 (2007).</li></ol>

著者らは、上記のテストを含むチャールズリバー、診断テストや試薬の供給者、すべての従業員です。

実験室で作業する場合、安全性は常に問題になるはず。動物を扱う場合に適切な保護具を着用すると前と後の消毒薬を使用してワークステーションを清掃することを忘れないでください。これらのメソッドは、主に、すなわちを検討し、彼らはエキゾチックなまたは非常に稀寄生虫だけでなく、より一般的な蟯虫と毛皮のダニを検出できる場所に実験用げっ歯類のいずれかの寄生虫を見つけるために設計されています。彼らは他の種にも等しく適用可能であるが、野生のげっ歯類は、上記の方法で評価されない、などの肝臓、皮下組織、脳などの場所で追加の寄生虫があるかもしれません。

<table border="1"><tbody><tr><td> 試薬名 </td><td> 会社 </td><td> カタログ番号 </td><td> コメント </td></tr><tr><td>まな板</td><td>サーモエレクトロンコーポレーション</td><td>猫＃36114 </td><td></td></tr><tr><td>小さなはさみ</td><td> ROBOZ </td><td> RS - 5910、G204 </td><td> 23ミリメートルブレード、3.5&quot;長さ、ストレート</td></tr><tr><td>培地はさみ</td><td> ROBOZ </td><td> RS - 6808、G207 </td><td> 5&quot; </td></tr><tr><td>鉗子に湾曲した</td><td> ROBOZ </td><td> RS - 8254（M1/21004） </td><td> 4.5&quot;、鋸歯状、わずかな曲線</td></tr><tr><td>鉗子- Microdissecting </td><td> ROBOZ </td><td> RS - 5238 </td><td>ハドソン-（エーヴァルト） </td></tr><tr><td>鉗子 - 組織鉗子</td><td> ROBOZ </td><td> RS - 8160 </td><td>ラットの歯</td></tr><tr><td>金属プローブ</td><td> VWR </td><td>猫＃25778-000 </td><td></td></tr><tr><td>止血</td><td>有利</td><td> V97 - 48 </td><td></td></tr><tr><td>アプリケータースティック</td><td>ピューリタン</td><td> 6インチアプリケーター、参照番号807 </td><td></td></tr><tr><td>解剖顕微鏡</td><td>オリンポス</td><td> SZ51、ショット、ACE1 </td><td></td></tr><tr><td>ペトリ皿</td><td> VWR </td><td> 100ミリメートル、猫＃3401PDNL </td><td></td></tr><tr><td>カバースリップ</td><td> VWR </td><td>マイクロカバーガラス、猫＃48366から067 </td><td></td></tr><tr><td>スライド</td><td> VWR </td><td> VistaVisionの顕微鏡スライドの猫＃16004から368 </td><td></td></tr><tr><td>接種ループ</td><td> VWR </td><td>猫＃50815-040 </td><td></td></tr><tr><td>光顕微鏡</td><td>オリンポス</td><td>モデル＃BX41TF </td><td></td></tr><tr><td>実験室のオーブン</td><td>クインシーラボ株式会社</td><td>モデル10研究室のオーブン</td><td></td></tr><tr><td>遠心</td><td>ベックマンコールター</td><td>アレグラX - 12R </td><td></td></tr><tr><td>ボルテックス</td><td> VWR </td><td>ミニボルテックス</td><td></td></tr><tr><td>テストチューブ</td><td>キンブル=チェイス</td><td> 15ミリリットルディスポーザブル遠沈管、カタログ番号73790〜15 </td><td></td></tr><tr><td>カバースリップ</td><td> VWR </td><td>プラスチック製の顕微鏡のカバーガラス、22ミリメートル、カタログ番号48376から049 </td><td></td></tr><tr><td>ホワイトキャップ</td><td> VWR </td><td>猫＃60869-089 </td><td></td></tr><tr><td>ヨウ素</td><td>ローリー生化学研究所</td><td>猫＃SO - 364 </td><td></td></tr><tr><td>亜鉛硫酸</td><td>シグマアルドリッチ</td><td>猫＃1000917519、Z4750 - 500G </td><td></td></tr><tr><td>スクロース</td><td> Mallinckrodtケミカルズ</td><td>猫＃8360〜06 </td><td></td></tr><tr><td>フェノール</td><td> EMD </td><td>猫＃PX0510 - 1 </td><td></td></tr><tr><td>セロハンテープ</td><td>ステープルズ</td><td>目に見えないテープ、猫＃504712 </td><td></td></tr><tr><td>鉱油</td><td> Mallickrodt </td><td>猫＃6358 </td><td></td></tr></tbody></table>

diagnosis of ecto- and endoparasites in laboratory rats and mice

内部および外部寄生虫は、実験室のげっ歯類の施設で重要な心配の種である、と多くの研究施設は、いくつかの寄生動物を抱く。寄生虫の動物の調査に着手する前に、二つのことを考慮する必要があります。 One：何を使用すると、収集した情報で作られ、二つになります：どのテストが最も適切です。動物は寄生されていることを知ることは、施設を受け入れるが、動物の治療にして、治療の有効性を判断する必要がしばしばあることなのかもしれません。寄生虫は、ライブまたは安楽死させた動物から採取したサンプルを含む様々な技術を通じて、動物で検出されることがあります。 PCRは、寄生虫のいくつかのタイプのための高感度のテストを許可しているものの、歴史的に、最大の診断感度テストでは、動物の安楽死を必要とした。この記事はの検出エンドおよびマウスとラットにおける外部寄生虫のための手順を示しています。見つかった寄生虫の種類が異なりますが、同様の手順では、他のげっ歯類にも適用可能である。

Watch this Scientific Journal Video about Diagnosis of Ecto- and Endoparasites in Laboratory Rats and Mice at JoVE.com

Diagnosis of Ecto- and Endoparasites in Laboratory Rats and Mice

この記事ではendo -または外部寄生を検出するスクリーニングのラットとマウスの様々な手順を説明します。いくつかの診断アッセイが実証され、生きた動物での使用に適したものと動物の安楽死の後に使用されているもの両方。ラットとマウスの寄生虫の同定に役立つように写真が含まれます。

実験用ラットとマウスにおけるエクト - とEndoparasitesの診断

Internal and external parasites remain a significant concern in laboratory rodent facilities, and many research facilities harbor some parasitized ...

diagnosis-of-ecto-and-endoparasites-in-laboratory-rats-and-mice

Nanyang Technological University

Research

JoVE Journal

Immunology and Infection

41.1K ビュー.  Charles River. この記事ではendo -または外部寄生を検出するスクリーニングのラットとマウスの様々な手順を説明します。いくつかの診断アッセイが実証され、生きた動物での使用に適したものと動物の安楽死の後に使用されているもの両方。ラットとマウスの寄生虫の同定に役立つように写真が含まれます。

ビデオ: Diagnosis of Ecto- and Endoparasites in Laboratory Rats and Mice

Orthotopic Hind-Limb Transplantation in Rats

Composite tissue allotransplantation (CTA) now represents a valid therapeutic option after the loss of a hand, forearm or digits and has become a novel therapeutic entity in reconstructive surgery. However, long term high-dose multi-drug immunosuppressive therapy is required to ensure graft survival, bearing the risk of serious side effects which halters broader application. Further progression in this field may depend on better understanding of basic immunology and ischemia reperfusion injury in composite tissue grafts.
To date, orthotopic hind limb transplantation in rats has been the preferred rodent model for reconstructive transplantation (RT), however, it is an extremely demanding procedure that requires extraordinary microsurgical skills for reattachment of vasculature, bones, muscles and nerves.
We have introduced the vascular cuff anastomosis technique to this model, providing a rapid and reliable approach to rat hind limb transplantation. This technique simplifies and shortens the surgical procedure and enables surgeons with basic microsurgical experience to successfully perform the operation with high survival and low complication rates. The technique seems to be well suited for immunological as well as ischemia reperfusion injury (IRI) studies.

Here we describe the orthotopic rat hind-limb transplantation procedure, which seems to be the gold standard in vivo model for composite tissue allotransplantation research.

ラット同所性後肢移植

Composite tissue allotransplantation (CTA) now represents a valid therapeutic option after the loss of a hand, forearm or digits and has become a novel ...

免疫・感染学

Genotypic Inference of HIV-1 Tropism Using Population-based Sequencing of V3

Background: Prior to receiving a drug from CCR5-antagonist class in HIV therapy, a patient must undergo an HIV tropism test to confirm that his or her viral population uses the CCR5 coreceptor for cellular entry, and not an alternative coreceptor. One approach to tropism testing is to examine the sequence of the V3 region of the HIV envelope, which interacts with the coreceptor.
Methods: Viral RNA is extracted from blood plasma. The V3 region is amplified in triplicate with nested reverse transcriptase-PCR. The amplifications are then sequenced and analyzed using the software, RE_Call. Sequences are then submitted to a bioinformatic algorithm such as geno2pheno to infer viral tropism from the V3 region. Sequences are inferred to be non-R5 if their geno2pheno false positive rate falls below 5.75%. If any one of the three sequences from a sample is inferred to be non-R5, the patient is unlikely to respond to a CCR5-antagonist.

HIV tropism can be inferred from the V3 region of the viral envelope. V3 is PCR amplified in triplicate using nested RT-PCR, sequenced, and interpreted using bioinformatic software. Samples with with 1 or more sequence(s) with low g2P scores are classified as non-R5 virus.

V3の人口ベースのシーケンシングを用いたHIV - 1トロピズムの遺伝子型推論

Background: Prior to receiving a drug from CCR5-antagonist class in HIV therapy, a patient must undergo an HIV tropism test to confirm that his or her ...

Basophil Activation Test for Investigation of IgE-Mediated Mechanisms in Drug Hypersensitivity

Hypersensitivity reactions against non-steroidal anti-inflammatory drugs (NSAIDs) like propyphenazone (PP) and diclofenac (DF) can manifest as Type I-like allergic reactions 1. In clinical practice, diagnosis of drug hypersensitivity is mainly performed by patient history, as skin testing is not reliable and oral provocation testing bears life-threatening risks for the patient 2. Hence, evidence for an underlying IgE-mediated pathomechanism is hard to obtain. 
Here, we present an in vitro method based on the use of human basophils derived from drug-hypersensitive patients that mimics the allergic effector reaction in vivo. As basophils of drug-allergic patients carry IgE molecules specific for the culprit drug, they become activated upon IgE receptor crosslinking and release allergic effector molecules. The activation of basophils can be monitored by the determination of the upregulation of CD63 surface expression using flow cytometry 3. 
In the case of low molecular weight drugs, conjugates are designed to enable IgE receptor crosslinking on basophils. As depicted in Figure 1, two representatives of NSAIDs, PP and DF, are covalently bound to human serum albumin (HSA) via a carboxyl group reacting with the primary amino group of lysine residues. DF carries an intrinsic carboxyl group and, thus, can be used directly 4, whereas a carboxyl group-containing derivative of PP had to be organochemically synthesized prior to the study 1.
The coupling degree of the low molecular weight compounds on the protein carrier molecule and their spatial distribution is important to guarantee crosslinking of two IgE receptor molecules. The here described protocol applies high performance-size exclusion chromatography (HPSEC) equipped with a sequential refractive index (RI) and ultra violet (UV) detection system for determination of the coupling degree.
As the described methodology may be applied for other drugs, the basophil activation test (BAT) bears the potential to be used for the determination of IgE-mediated mechanisms in drug hypersensitivity. Here, we determine PP hypersensitivity as IgE-mediated and DF hypersensitivity as non-IgE-mediated by BAT.

Basophil activation test is a potent tool for the detection of IgE-dependent allergies in vitro. Here, an optimized protocol for basophil activation test is used to investigate drug hypersensitivity. A method for the efficient production of covalent drug-protein conjugates and their physicochemical characterization is described.

薬物過敏症におけるIgEが介在する機序の解明のための好塩基球の活性化試験

Hypersensitivity reactions against non-steroidal anti-inflammatory drugs (NSAIDs) like propyphenazone (PP) and diclofenac (DF) can manifest as Type I-like ...

Evaluation of Zika Virus-specific T-cell Responses in Immunoprivileged Organs of Infected Ifnar1-/- Mice

The Zika virus (ZIKV) can induce inflammation in immunoprivileged organs (e.g., the brain and testis), leading to the Guillain-Barr&#233; syndrome and damaging the testes. During an infection with the ZIKV, immune cells have been shown to infiltrate into the tissues. However, the cellular mechanisms that define the protection and/or immunopathogenesis of these immune cells during a ZIKV infection are still largely unknown. Herein, we describe methods to evaluate the virus-specific T-cell functionality in these immunoprivileged organs of ZIKV-infected mice. These methods include a) a ZIKV infection and vaccine inoculation in Ifnar1-/- mice; b) histopathology, immunofluorescence, and immunohistochemistry assays to detect the virus infection and inflammation in the brain, testes, and spleen; c) the preparation of a tetramer of ZIKV-derived T-cell epitopes; d) the detection of ZIKV-specific T cells in the monocytes isolated from the brain, testes, and spleen. Using these approaches, it is possible to detect the antigen-specific T cells that have infiltrated into the immunoprivileged organs and to evaluate the functions of these T cells during the infection: potential immune protection via virus clearance and/or immunopathogenesis to exacerbate the inflammation. These findings may also help to clarify the contribution of T cells induced by the immunization against ZIKV.

Evaluation of Zika Virus-specific T-cell Responses in Immunoprivileged Organs of Infected Ifnar1-/- Mice

A protocol to evaluate antigen-specific T-cell responses in the immunoprivileged organs of the Ifnar1-/- murine model for the Zika virus (ZIKV) infection is described. This method is pivotal for investigating the cellular mechanisms of the protection and immunopathogenesis of ZIKV vaccines and is also valuable for their efficacy evaluation.

ジカ ウイルス特異 Immunoprivileged 臓器感染 Ifnar1-/-マウスの T 細胞応答の評価

The Zika virus (ZIKV) can induce inflammation in immunoprivileged organs (e.g., the brain and testis), leading to the Guillain-Barr&#233; syndrome and ...

Quantification of Atherosclerosis in Mice

Cardiovascular disease is the main cause of death in the world. The underlying cause in most cases is atherosclerosis, which is in part a chronic inflammatory disease. Experimental atherosclerosis studies have elucidated the role of cholesterol and inflammation in the disease process. This has led to successful clinical trials with pharmaceutical agents that reduce clinical manifestations of atherosclerosis. Careful and well-controlled experiments in mouse models of the disease could further elucidate the pathogenesis of the disease, which is not fully understood. Standardized lesion analysis is important to reduce experimental variability and increase reproducibility. Determining lesion size in aortic root, aortic arch, and brachiocephalic artery are common endpoints in experimental atherosclerosis. This protocol provides a technical description for evaluation of atherosclerosis at all these sites in a single mouse. The protocol is particularly useful when material is limited, as is frequently the case when genetically modified animals are being characterized.

Murine models of atherosclerosis are useful tools to investigate pathogenic pathways on a molecular level, but require standardized quantification of lesion development. This protocol describes an optimized method to determine lesion size in the major arterial vessels including the aortic root, aortic arch, and brachiocephalic artery.

マウスにおけるアテローム性動脈硬化症の定量化

Cardiovascular disease is the main cause of death in the world. The underlying cause in most cases is atherosclerosis, which is in part a chronic ...

Adoptive Immunotherapy of iNKT Cells in Glucose-6-Phosphate Isomerase (G6PI)-Induced RA Mice

Rheumatoid arthritis (RA) is a complex chronic inflammatory autoimmune disease. The pathogenesis of the disease is related to invariant natural killer T (iNKT) cells. Patients with active RA present fewer iNKT cells, defective cell function, and excessive polarization of Th1. In this study, an RA animal model was established using a mixture of hGPI325-339 and hGPI469-483 peptides. The iNKT cells were obtained by in vivo induction and in vitro purification, followed by infusion into RA mice for adoptive immunotherapy. The in vivo imaging system (IVIS) tracking revealed that iNKT cells were mainly distributed in the spleen and liver. On day 12 after cell therapy, the disease progression slowed down significantly, the clinical symptoms were alleviated, the abundance of iNKT cells in the thymus increased, the proportion of iNKT1 in the thymus decreased, and the levels of TNF-&#945;, IFN-&#947;, and IL-6 in the serum decreased. Adoptive immunotherapy of iNKT cells restored the balance of immune cells and corrected the excessive inflammation of the body.

Adoptive Immunotherapy of iNKT Cells in Glucose-6-Phosphate Isomerase G6PI-Induced RA Mice

This protocol uses G6PI mixed peptides to construct rheumatoid arthritis models that are closer to that of human rheumatoid arthritis in CD4+ T cells and cytokines. High purity invariant natural killer T cells (mainly iNKT2) with specific phenotypes and functions were obtained by in vivo induction and in vitro purification for adoptive immunotherapy.

グルコース-6-リン酸イソメラーゼ(G6PI)誘発RAマウスにおけるiNKT細胞の導入免疫療法

Rheumatoid arthritis (RA) is a complex chronic inflammatory autoimmune disease. The pathogenesis of the disease is related to invariant natural killer T ...

Evaluation of T Follicular Helper Cells and Germinal Center Response During Influenza A Virus Infection in Mice

T Follicular Helper (Tfh) cells are an independent CD4+&#160;T cell subset specialized in providing help for germinal center (GC) development and generation of high-affinity antibodies. In influenza virus infection, robust Tfh and GC B cell responses are induced to facilitate effective virus eradication, which confers a qualified mouse model for Tfh-associated study. In this paper, we described protocols in detection of basic Tfh-associated immune response during influenza virus infection in mice. These protocols include: intranasal inoculation of influenza virus; flow cytometry staining and analysis of polyclonal and antigen-specific Tfh cells, GC B cells and plasma cells; immunofluorescence detection of GCs; enzyme-linked immunosorbent assay (ELISA) of influenza virus-specific antibody in serum. These assays basically quantify the differentiation and function of Tfh cells in influenza virus infection, thus providing help for studies in elucidating differentiation mechanism and manipulation strategy.

This paper describes protocols of evaluating Tfh and GC B response in mouse model of influenza virus infection.

マウスにおけるインフルエンザAウイルス感染時のT濾胞ヘルパー細胞と胚葉センター応答の評価

T Follicular Helper (Tfh) cells are an independent CD4+&#160;T cell subset specialized in providing help for germinal center (GC) development and ...

Conjunctival Commensal Isolation and Identification in Mice

The ocular surface was once considered immune privileged and abiotic, but recently it appears that there is a small, but persistent commensal presence. Identification and monitoring of bacterial species at the ocular mucosa have been challenging due to their low abundance and limited availability of appropriate methodology for commensal growth and identification. There are two standard approaches: culture based or DNA sequencing methods. The first method is problematic due to the limited recoverable bacteria and the second approach identifies both live and dead bacteria leading to an aberrant representation of the ocular space. We developed a robust and sensitive method for bacterial isolation by building upon standard microbiological culturing techniques. This is a swab-based technique, utilizing an &#8220;in-lab&#8221; made thin swab that targets the lower conjunctiva, followed by an amplification step for aerobic and facultative anaerobic genera. This protocol has allowed us to isolate and identify conjunctival species such as Corynebacterium spp., Coagulase Negative Staphylococcus spp., Streptococcus spp., etc. The approach is suitable to define commensal diversity in mice under different disease conditions.

Presented here is a protocol for the isolation and amplification of aerobic and facultative anaerobic mouse conjunctival commensal bacteria using a unique eye swab and culture-based enrichment step with subsequent identification by microbiological based methods and MALDI-TOF mass spectrometry.

マウスにおける結膜経月の分離と同定

The ocular surface was once considered immune privileged and abiotic, but recently it appears that there is a small, but persistent commensal presence. ...

Recurrent Escherichia coli Urinary Tract Infection Triggered by Gardnerella vaginalis Bladder Exposure in Mice

Recurrent urinary tract infections (rUTI) caused by uropathogenic Escherichia coli (UPEC) are common and costly. Previous articles describing models of UTI in male and female mice have illustrated the procedures for bacterial inoculation and enumeration in urine and tissues. During an initial bladder infection in C57BL/6 mice, UPEC establish latent reservoirs inside bladder epithelial cells that persist following clearance of UPEC bacteriuria. This model builds on these studies to examine rUTI caused by the emergence of UPEC from within latent bladder reservoirs. The urogenital bacterium Gardnerella vaginalis is used as the trigger of rUTI in this model because it is frequently present in the urogenital tracts of women, especially in the context of vaginal dysbiosis that has been associated with UTI. In addition, a method for in situ bladder fixation followed by scanning electron microscopy (SEM) analysis of bladder tissue is also described, with potential application to other studies involving the bladder.

Recurrent Escherichia coli Urinary Tract Infection Triggered by Gardnerella vaginalis Bladder Exposure in Mice

A mouse model of uropathogenic E. coli (UPEC) transurethral inoculation to establish latent intracellular bladder reservoirs and subsequent bladder exposure to G. vaginalis to induce recurrent UPEC UTI is demonstrated. Also demonstrated are the enumeration of bacteria, urine cytology, and in situ bladder fixation and processing for scanning electron microscopy.

マウスにおけるガードネレラ膣膀胱曝露によって引き起こされる再発性大腸菌尿路感染症

Recurrent urinary tract infections (rUTI) caused by uropathogenic Escherichia coli (UPEC) are common and costly. Previous articles describing models of ...

Partial Heterotopic Hindlimb Transplantation Model in Rats

Vascularized composite allotransplantations (VCA) represent the most advanced reconstruction option for patients without autologous surgical possibilities after a complex tissue defect. Face and hand transplantations have changed disfigured patients' lives, giving them a new aesthetic and functional social organ. Despite promising outcomes, VCA is still underperformed due to life-long immunosuppression comorbidities and infectious complications. The rat is an ideal animal model for in vivo studies investigating immunological pathways and graft rejection mechanisms. Rats are also widely used in novel composite tissue graft preservation techniques, including perfusion and cryopreservation studies. Models used for VCA in rats must be reproducible, reliable, and efficient with low postoperative morbidity and mortality. Heterotopic limb transplantation procedures fulfill these criteria and are easier to perform than orthotopic limb transplants. Mastering rodent microsurgical models requires solid experience in microsurgery and animal care. Herein is reported a reliable and reproducible model of partial heterotopic osteomyocutaneous flap transplantation in rats, the postoperative outcomes, and the means of prevention of potential complications.

This paper presents a partial heterotopic osteomyocutaneous flap transplantation protocol in rats and its potential outcomes in the mid-term follow-up.

ラットにおける部分的な異所性後肢移植モデル

Vascularized composite allotransplantations (VCA) represent the most advanced reconstruction option for patients without autologous surgical possibilities ...