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A Comparison of LED with Fluorescent Lighting on the Stress, Behavior, and Reproductive Success of Laboratory Zebra Finches (Taeniopygia guttata). LED 与荧光灯对实验室斑马雀(Taeniopygia guttata)的压力、行为和繁殖成功率的影响比较。
Pub Date : 2024-05-01 Epub Date: 2024-04-29 DOI: 10.30802/AALAS-JAALAS-24-000009
Alanna G Backx, April Wu, Alyx Tanner, Niora J Fabian

There are limited evidence-based husbandry recommendations for laboratory zebra finches (Taeniopygia guttata), including appropriate light sources. Light-emitting diode (LED) technology has been shown to improve circadian regulation and reduce stress in some laboratory animal species, such as mice and rats, when compared with cool-white fluorescent (CWF) lighting, but the effects of LED lighting on zebra finches have not been published. We compared the effects of broad-spectrum, blue-enriched (6,500 Kelvin) CWF and flicker-free LED lighting on the behavior, stress, and reproductive outcomes of indoor-housed zebra finches. Using breeding pairs housed in cubicles illuminated with either CWF or LED lighting, we compared the reproductive output as determined by clutch size, hatching rate, and hatchling survival rate. We also compared the behavior of group-housed adult males, first housed under CWF followed by LED lighting, using video recordings and an ethogram. Fecal samples were collected from these males at the end of each recording period, and basal fecal corticosterone metabolite (FCM) levels were compared. A FCM assay for adult male zebra finches was validated for efficacy and accuracy using a capture-restraint acute stress response and parallelism analysis, respectively. The breeding pairs had no significant difference in the clutch size or percent hatching rate, but percent hatchling survival improved under LED with an increased proportion achieving 100% survival. There was no significant difference in FCM between the lighting treatments. However, the activity budgets of the birds were altered, with a reduction in flighted movement and an increase in enrichment manipulation under LED. Overall, these results support the use of blue-enriched, broad-spectrum flicker-free LED as a safe alternative to CWF lighting for breeding and nonbreeding indoor-housed zebra finches.

目前针对实验室斑马雀(Taeniopygia guttata)的饲养建议(包括适当的光源)证据有限。与冷白荧光(CWF)照明相比,发光二极管(LED)技术已被证明可改善昼夜节律调节,并减少小鼠和大鼠等一些实验室动物物种的压力,但 LED 照明对斑马雀的影响尚未公布。我们比较了广谱、富蓝(6500开尔文)CWF和无闪烁LED照明对室内饲养斑马雀的行为、压力和繁殖结果的影响。我们利用饲养在使用CWF或LED照明的隔间中的斑马雀繁殖配对,比较了繁殖产量(由窝产仔数、孵化率和幼鸟存活率决定)。我们还使用视频记录和电子图谱对群居成年雄鸟的行为进行了比较。在每个记录期结束时收集这些雄鸟的粪便样本,并比较粪便中皮质酮代谢物(FCM)的基础水平。针对成年雄性斑马雀的粪皮质酮代谢物检测方法分别通过捕获-限制急性应激反应和平行分析验证了其有效性和准确性。配对繁殖的斑马雀在窝产仔数和孵化率方面没有显著差异,但在发光二极管的作用下,孵化成活率有所提高,100%成活的比例有所增加。照明处理之间的FCM没有明显差异。但是,鸟类的活动预算发生了变化,在 LED 下飞行运动减少,而强化操作增加。总之,这些结果支持使用富含蓝色的宽光谱无闪烁 LED,作为繁殖和非繁殖室内饲养斑马雀的 CWF 照明的安全替代品。
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引用次数: 0
Comparison of Plenum and Cage-level Filter Exhaust Dust PCR Testing to Soiled Bedding Sentinel Mice (Mus musculus) on an IVC Rack. 对 IVC 架上脏污垫料哨兵小鼠(Mus musculus)进行全空气和笼级过滤器排气粉尘 PCR 测试的比较。
Pub Date : 2024-05-01 Epub Date: 2024-03-01 DOI: 10.30802/AALAS-JAALAS-23-000073
Wendy R Williams, Shawn P Lane, Cheryl Perkins, Ken Henderson

The use of soiled-bedded sentinels (SBSs) has historically been the standard for colony health surveillance monitoring at our institution. With the advent of newer technologies in which dust collected from filters is tested by PCR, we compared traditional SBS with PCR testing of both exhaust air dust collected from a filter in the downstream vertical plenum (exhaust dust test [EDT]) and the SBS cage-level exhaust filter (SCEF). Our hypothesis was that both methods of filter testing would identify more pathogens than SBS testing. Twenty-five individually ventilated mouse racks that used disposable caging were sanitized and placed into rotation. Rack plenums were tested by PCR to verify negative results before the study start. Exhaust dust collection media were placed in the exhaust plenum (n = 25). SBS cages were placed on each side of the rack with 2 mice per cage (n = 42 mice), with the remaining cage slots occupied by research animals. At each triweekly cage change, the exhaust air filters were carefully removed from the cage top, placed in sterile 50-mL conical tubes, and pooled for submission. After 3mo, the SBS mice were tested via serology for bacterial and viral agents and by PCR for Helicobacter species, pinworms, and ectoparasites. In addition, the EDT filter and SCEF were collected for PCR to evaluate for the same agents. Our results indicate that the SCEF consistently detected agents more frequently than the EDT filter placed in the plenum and that the EDT filter media detected agents more frequently than did the SBS mice. Our data suggest that both PCR methods of detection are superior to SBS for individually ventilated disposable rodent cages and that the SCEF is superior to EDT. These data supported our movement of institution toward environmental monitoring as a method of rodent colony health surveillance.

在我们机构,使用污床哨兵(SBS)一直是菌落健康监测的标准。随着从过滤器中收集的粉尘通过 PCR 进行检测的新技术的出现,我们将传统的 SBS 与从下游垂直管道中的过滤器收集的废气粉尘(排气粉尘检测 [EDT])和 SBS 笼级排气过滤器(SCEF)的 PCR 检测进行了比较。我们的假设是,这两种过滤器检测方法都能比 SBS 检测方法识别出更多的病原体。我们对 25 个使用一次性笼具的独立通风小鼠架进行了消毒,并将其轮流放置。在研究开始前,用 PCR 对鼠架管道进行检测,以验证阴性结果。排气集尘介质被放置在排气管中(n = 25)。SBS 笼放置在机架两侧,每笼 2 只小鼠(n = 42 只),其余笼位由研究动物占据。每次三周换笼时,从笼子顶部小心取出排气过滤器,放入无菌的 50 毫升锥形试管中,然后集中送检。3 mo 后,SBS 小鼠通过血清学检测细菌和病毒,并通过 PCR 检测螺旋杆菌、蛲虫和体外寄生虫。此外,还收集了 EDT 过滤器和 SCEF 进行 PCR 检测,以评估相同的病原体。我们的结果表明,SCEF 检测到病原体的频率始终高于放置在通风孔中的 EDT 过滤器,EDT 过滤介质检测到病原体的频率高于 SBS 小鼠。我们的数据表明,对于单独通风的一次性啮齿动物笼而言,两种 PCR 检测方法都优于 SBS,而且 SCEF 优于 EDT。这些数据支持我们的机构将环境监测作为啮齿动物群落健康监测的一种方法。
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引用次数: 0
Noise as an Extrinsic Variable in the Animal Research Facility. 噪音是动物研究设施中的一个外部变量。
Pub Date : 2024-05-01 Epub Date: 2024-05-15 DOI: 10.30802/AALAS-JAALAS-24-000008
Jeremy G Turner, John R Manker

Animal research facilities are noisy environments. The high air change rates required in animal housing spaces tend to create higher noise levels from the heating and cooling systems. Housing rooms are typically constructed of hard wall material that is easily cleaned but simultaneously highly reverberant, meaning that the sound cannot be controlled/attenuated so the sounds that are generated bounce around the room uncontrolled. (Soft, sound-absorbing surfaces generally cannot be used in animal facilities because they collect microbes; various wall surface features and sound control panel options are available, although rarely used.) In addition, many of our husbandry tasks such as cage changing, animal health checks, cleaning, and transporting animals produce high levels of noise. Finally, much of the equipment we have increasingly employed in animal housing spaces, such as ventilated caging motors, biosafety, or procedure cabinets, can generate high levels of background noise, including ultrasound. These and many additional factors conspire to create an acoustic environment that is neither naturalistic nor conducive to a stress-free environment. The acoustic variability both within and between institutions can serve as an enormous confounder for research studies and a threat to our ability to reproduce studies over time and between research laboratories. By gaining a better appreciation for the acoustic variables, paired with transparency in reporting the levels, we might be able to gain a better understanding of their impacts and thereby gain some level of control over such acoustic variables in the animal housing space. The result of this could improve both animal welfare and study reproducibility, helping to address our 3Rs goals of Replacement, Reduction, and Refinement in the animal biomedical research enterprise.

动物研究设施环境嘈杂。动物饲养空间所需的高换气率往往会导致供暖和制冷系统产生更高的噪音。饲养室通常使用硬质墙壁材料,这种材料易于清洁,但同时具有高混响性,这意味着声音无法控制/衰减,因此产生的声音会在室内不受控制地反弹。(柔软的吸音表面一般不能用于动物设施,因为它们会聚集微生物;有各种墙面特征和声音控制面板可供选择,但很少使用)。此外,我们的许多饲养工作,如换笼、动物健康检查、清洁和运输动物等,都会产生大量噪音。最后,我们在动物饲养空间越来越多地使用的许多设备,如通风笼电机、生物安全柜或程序柜,都会产生高水平的背景噪声,包括超声波。这些因素和许多其他因素共同造成了一种既不自然也不利于营造无压力环境的声学环境。机构内部和机构之间的声学差异会对研究产生巨大的干扰,并威胁到我们在不同时间和不同研究实验室之间重现研究的能力。通过更好地了解声学变量,再加上透明的报告水平,我们或许能更好地理解它们的影响,从而在一定程度上控制动物饲养空间中的声学变量。这样做的结果既能改善动物福利,又能提高研究的可重复性,有助于在动物生物医学研究事业中实现 "替代、减少和完善 "的 3R 目标。
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引用次数: 0
Evaluating a Reduction in Treatment Duration of Ivermectin Diet for Fur Mite (Radfordia affinis) Eradication in Mice. 评估缩短伊维菌素饮食治疗小鼠毛螨(Radfordia affinis)的持续时间。
Pub Date : 2024-04-25 DOI: 10.30802/AALAS-JAALAS-24-000007
Wai H Hanson Dvm PhD Daclam, Cayden J Samuels Ba, Cheryl L Woods Bs, Kenneth S Henderson PhD MSc
Murine fur mites are commonly excluded in modern research animal programs, yet infestations continue to persist due to challenges in detection and control. Because all diagnostic methods and treatment options have limitations, programs must make many operational decisions when trying to eradicate these ectoparasites. The primary aim of this study was to assess various durations of treatment time with an ivermectin-compounded diet in eliminating Radfordia affinis in mice as determined by PCR testing and pelt examination. A shorter treatment duration would be highly advantageous as compared with the current regimen of 8 wk as it would minimize cost and time for animal management programs, impediments to research, and ivermectin drug effects on infested animals. Five experimental groups of R. affinis-positive mice received dietary ivermectin for 0, 2, 4, 6, or 8 wk. A fur mite-negative, naïve mouse was added to each group every 8 wk to perpetuate the infestation and amplify any remaining populations of fur mites. At 16 wk after the respective treatment end, PCR testing was performed for all treated groups in conjunction with the positive control group (no treatment). Visual examination of pelts for mites and eggs via direct microscopy was also performed at each time point. All treated mice were free of R. affinis at 16 wk after the end of treatment as confirmed by both PCR testing and pelt examination. These findings indicate that a dietary ivermectin treatment duration of as little as 2 wk is effective in eliminating R. affinis, making successful eradication initiatives more achievable.
在现代研究动物项目中,鼠毛螨通常被排除在外,但由于检测和控制方面的挑战,鼠毛螨的侵扰仍然持续存在。由于所有诊断方法和治疗方案都有局限性,因此项目在试图根除这些体外寄生虫时必须做出许多操作决定。本研究的主要目的是评估伊维菌素复配饲料在消除小鼠体内 Radfordia affinis(通过 PCR 检测和毛皮检查确定)方面的各种治疗持续时间。与目前的 8 周治疗方案相比,缩短治疗时间将非常有利,因为这将最大限度地减少动物管理项目的成本和时间、对研究的阻碍以及伊维菌素药物对受感染动物的影响。五个实验组的 R. affinis 阳性小鼠分别在 0、2、4、6 或 8 周内服用伊维菌素。每隔 8 周,每组中加入一只毛螨阴性的天真小鼠,以延续感染并扩大剩余的毛螨种群。治疗结束 16 周后,对所有治疗组和阳性对照组(无治疗)进行 PCR 检测。在每个时间点,还通过直接显微镜目测毛皮上的螨虫和虫卵。经 PCR 检测和毛皮检查证实,所有治疗小鼠在治疗结束后 16 周内均未发现 R. affinis。这些研究结果表明,只需 2 周的伊维菌素饮食治疗时间就能有效消灭 R. affinis,从而使根除行动更容易取得成功。
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引用次数: 0
Opinion: A Seven-step Approach to Communication about Animal Research. 意见:动物研究沟通七步法》。
Pub Date : 2024-04-25 DOI: 10.30802/AALAS-JAALAS-23-000108
Eric P Sandgren Vmd PhD
Organizations that receive public money to conduct research using animals should be able to explain the importance of and need for that work. More generally, anyone who believes that properly conducted and regulated animal research either does or does not make the world a better place wants the public to understand why they hold their belief. In a world with divided support for animal research, honest communication about these issues is essential to develop sound public policy. Specifically, communication about animal research (or any type of research) needs to address the scientific, ethical, and regulatory considerations that underlie public policy decisions. This opinion article describes a 7-step communication strategy designed to address these issues. The 7 elements of this approach are 1) motivation, 2) the right mix of information, 3) a team approach, 4) respect for your audience, 5) determination and courage, 6) humility and honesty, and 7) persistence.
接受公共资金利用动物进行研究的组织应该能够解释这项工作的重要性和必要性。更广泛地说,任何认为适当开展和监管动物研究能够或不能使世界变得更美好的人,都希望公众了解他们为何坚持自己的信念。在这个对动物研究的支持存在分歧的世界里,就这些问题进行坦诚的沟通对于制定合理的公共政策至关重要。具体而言,有关动物研究(或任何类型的研究)的沟通需要涉及作为公共政策决策基础的科学、伦理和监管因素。这篇观点文章介绍了旨在解决这些问题的七步沟通策略。该方法的 7 个要素是:1)动机;2)正确的信息组合;3)团队方法;4)尊重受众;5)决心和勇气;6)谦逊和诚实;7)坚持不懈。
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引用次数: 0
Association of Primate Veterinarians Guidelines for Cerebrospinal Fluid Aspiration for Nonhuman Primates in Biomedical Research. 灵长类动物兽医协会生物医学研究非人灵长类动物脑脊液抽吸指南。
Pub Date : 2024-04-23 DOI: 10.30802/AALAS-JAALAS-24-000029
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引用次数: 0
An Epidemiologic Study of Bacterial Culture and Antibiotic Susceptibility Analyses in Captive Macaques and Marmosets at the Wisconsin National Primate Research Center. 威斯康星国家灵长类动物研究中心对圈养猕猴和狨猴进行细菌培养和抗生素敏感性分析的流行病学研究。
Pub Date : 2024-04-22 DOI: 10.30802/AALAS-JAALAS-23-000079
Emma L Svenson, J. Coonen, James E Svenson, Heather A Simmons, Jennifer M Hayes, Saverio Capuano
Antimicrobial resistance (AMR) represents a growing public health threat that arises at the interface between animal, human, and environmental health. Although the pathways promoting the development of AMR are well characterized in human health settings, data within the veterinary medical world are less abundant, particularly from fields focusing on nontraditional species, such as nonhuman primates (NHPs). The purpose of this study was to describe trends in sample submission for bacterial culture, characterize patterns of microbial growth and any changes in AMR and susceptibility over time, and inform best practices for veterinary antimicrobial stewardship in a captively-housed, indoor NHP colony. Electronic health records from the Wisconsin National Primate Research Center were analyzed across a 10-y period using SAS Studio. There was an increasing pattern of sample submissions for culture and susceptibility analyses, with no corresponding increases in resistance to relevant antibiotics for potential zoonotic pathogens, such as Escherichia coli or Shigella species. Trends are suggestive of appropriate antimicrobial stewardship practices that were responsive to the medical needs of Wisconsin National Primate Research Center animals, as well as the needs of the larger research community at the University of Wisconsin-Madison. These findings can inform veterinary professionals working with NHPs and contribute to the growing body of literature surrounding AMR in nontraditional species.
抗菌药耐药性(AMR)是一个日益严重的公共卫生威胁,它出现在动物、人类和环境健康的交界处。虽然人类健康环境中促进 AMR 发展的途径已被充分描述,但兽医领域的数据却不那么丰富,尤其是来自非传统物种领域的数据,如非人灵长类动物 (NHP)。本研究的目的是描述细菌培养样本提交的趋势,描述微生物生长的模式以及随着时间推移 AMR 和敏感性的任何变化,并为圈养的室内 NHP 群落中兽医抗菌药管理的最佳实践提供信息。我们使用 SAS Studio 分析了威斯康星国家灵长类动物研究中心 10 年间的电子健康记录。提交的培养和药敏分析样本数量呈上升趋势,而潜在人畜共患病病原体(如大肠杆菌或志贺氏杆菌)对相关抗生素的耐药性却没有相应增加。这些趋势表明,威斯康星国家灵长类动物研究中心采取了适当的抗菌药物管理措施,以满足动物的医疗需求以及威斯康星大学麦迪逊分校广大研究社区的需求。这些研究结果可以为从事 NHPs 工作的兽医专业人员提供参考,并为围绕非传统物种 AMR 的不断增加的文献做出贡献。
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引用次数: 0
Comparing Genotyping Accuracy Using Buccal Swabs versus Tail Biopsies by PCR in B6;C3-Tg(Prnp-SNCA*A53T)83Vle and B6;C3-Tg(Prnp-SNCA*A53T)83Vle Sncatm1Mjff Mice. 通过 PCR 比较 B6;C3-Tg(Prnp-SNCA*A53T)83Vle 和 B6;C3-Tg(Prnp-SNCA*A53T)83Vle Sncatm1Mjff 小鼠口腔拭子与尾部活检的基因分型准确性。
Pub Date : 2024-04-22 DOI: 10.30802/AALAS-JAALAS-23-000045
Ming F Lui Dvm, Melissa Osborne Ms, Todd Dehm Ma, Min Lee Ba, Julian A Castaneda Dvm PhD Daclam
Genotyping is a common and necessary procedure performed on genetically modified animals to distinguish carriers from noncarriers of the variants of interest. Established methods involve collection of tissues such as tips of tails or notches of ears. Noninvasive methods have been described but not widely adopted for reasons including inertia to change, needs to adjust PCR protocols, and the lack of validation; noninvasive genotyping methods are a refinement on animal welfare, but questions remain regarding how they compare with invasive methods in terms of genotyping accuracy rate and reproducibility. To gain answers to these questions, we compared the detection accuracy of the transgene and determination of zygosity in B6;C3-Tg(Prnp-SNCA*A53T)83Vle and B6;C3-Tg(Prnp-SNCA*A53T)83Vle Sncatm1Mjff neonatal mice between tail biopsies and buccal swabs. Moreover, we weighed and observed mice following genotyping to see if any clinical differences can be discerned. Weight data did not support statistically significant differences in mice undergoing different genotyping procedures and control. No statistically significant difference was found between using buccal swabs or tail biopsies for genotyping with PCR or quantitative PCR. None of the pups swabbed was rejected by the dam. Our findings indicate that buccal swabbing is a more humane and feasible alternative to tail biopsies for high-throughput genotyping.
基因分型是对转基因动物进行基因分型的一种常见且必要的程序,用于区分相关变异体的携带者和非携带者。现有的方法需要采集组织,如尾巴尖或耳朵的缺口。无创基因分型方法是对动物福利的一种改进,但在基因分型准确率和可重复性方面与有创方法的比较仍存在问题。为了回答这些问题,我们比较了 B6;C3-Tg(Prnp-SNCA*A53T)83Vle和 B6;C3-Tg(Prnp-SNCA*A53T)83Vle Sncatm1Mjff 新生小鼠尾部活检与颊拭子之间的转基因检测准确性和子系测定。此外,我们还对基因分型后的小鼠进行了称重和观察,以确定是否存在临床差异。不同基因分型程序和对照组小鼠的体重数据在统计学上没有显著差异。使用口腔拭子或尾部活检进行 PCR 或定量 PCR 基因分型在统计学上没有明显差异。没有一只幼鼠被母鼠排斥。我们的研究结果表明,在进行高通量基因分型时,颊拭子比尾巴活检更人道、更可行。
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引用次数: 0
Effect of Adopting a Timothy Hay-based Diet at Weaning or in Adulthood on Urinary Tract Parameters in Strain 13/N Guinea Pigs (Cavia porcellus). 断奶期或成年期采用以提摩西干草为主的日粮对13/N品系豚鼠(Cavia porcellus)尿路参数的影响
Pub Date : 2024-04-05 DOI: 10.30802/AALAS-JAALAS-24-000019
Rachel C Wier, Timothy D. Flietstra, J. Coleman-McCray, S. Genzer, Marie E Brake, Eric M Velazquez, Catalina Forero, S. R. Welch, Cassandra M Tansey, Jillian A. Condrey, J. Spengler
Type of feed is an important consideration in herbivore colony management, yet limited studies report on the effects of diet on common conditions such as urolithiasis in guinea pigs. Urolithiasis is a well-documented cause of lower urinary tract disease in guinea pigs, with calcium carbonate uroliths reported as the predominant calculi formed in the guinea pig urinary tract. A calcium-rich diet has been suggested as a risk factor for of urolithiasis, with numerous commercially available guinea pig diets formulated for adults avoiding ingredients that are higher in calcium. Due to the high incidence of urolithiasis in our strain 13/N guinea pig colony, we conducted a prospective control study following the implementation of dietary changes aimed at improving overall urinary tract health and reducing risk factors for urolithiasis, thus improving colony welfare. A control group was kept on the original ad libitum alfalfa hay-based pellet diet with restricted loose timothy hay (control diet, 14 juveniles and 24 adults). An experimental group was placed on a portioned, 1 oz daily, timothy hay-based pellet diet with ad libitum loose timothy hay (experimental diet, 21 juveniles and 23 adults). Juveniles and adults were followed for a total of 14 and 26 wk, respectively. Longitudinal blood and urine samples were collected to evaluate blood chemistry and urinary parameters, along with weight and body condition scores to assess general health. Overall, dietary changes did not improve parameters associated with improved urinary tract health or reduced risk of urolithiasis; feeding strategy was not found to meaningfully affect calcium crystalluria, urine protein, urine specific gravity, or renal values. These data support alfalfa hay-based pellet or timothy hay-based pellet, when fed with loose timothy hay, as viable options and suggest that practices aimed at reducing dietary calcium by reducing pelleted diet portions are insufficient to mitigate risk factors for urolithiasis in guinea pigs.
饲料类型是食草动物群落管理的一个重要考虑因素,但有关饮食对豚鼠尿石症等常见疾病的影响的研究报告却很有限。尿路结石是豚鼠下尿路疾病的主要病因,据报道,碳酸钙尿路结石是豚鼠尿路中形成的主要结石。富含钙质的饮食被认为是导致豚鼠患尿路结石的一个危险因素,市面上许多为成年豚鼠配制的日粮都避免使用含钙量较高的成分。由于我们的 13/N 品系豚鼠群中尿路结石的发病率很高,因此我们进行了一项前瞻性对照研究,在实施饮食改变后,旨在改善整体尿路健康,减少尿路结石的风险因素,从而提高豚鼠群的福利。对照组采用原来的自由采食苜蓿干草颗粒饲料,并限制使用松散的提摩西干草(对照组 14 只幼鼠和 24 只成年鼠)。实验组每天喂食 1 盎司定量的提摩西干草颗粒饲料和自由采食的松散提摩西干草(实验组 21 只幼鼠和 23 只成鼠)。分别对幼鼠和成鼠进行了为期 14 周和 26 周的跟踪观察。收集纵向血液和尿液样本以评估血液化学和尿液参数,同时收集体重和身体状况评分以评估总体健康状况。总体而言,改变饮食并没有改善与改善尿路健康或降低尿路结石风险相关的参数;饲喂策略也没有对钙结晶尿、尿蛋白、尿比重或肾脏值产生有意义的影响。这些数据支持将苜蓿干草颗粒饲料或提摩西干草颗粒饲料与松散的提摩西干草一起饲喂作为可行的选择,并表明通过减少颗粒饲料的份量来降低膳食钙的做法不足以减轻豚鼠患尿路结石的风险因素。
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引用次数: 0
Polymerase Chain Reaction on In-cage Filter Paper at Different Time Points to Detect Helicobacter spp. 不同时间点的笼内滤纸聚合酶链式反应检测螺旋杆菌属
Pub Date : 2024-04-02 DOI: 10.30802/AALAS-JAALAS-24-000001
Abby C Bernardini, Wendy R Williams
Helicobacter spp. infections in mice can have broad-ranging effects on gastrointestinal, reproductive, and immune systems. This can introduce significant confounding variables for research and may reduce scientific rigor. Screening mouse colonies for Helicobacter species can be accomplished via noninvasive PCR testing on filter paper placed in animal-free dirty bedding sentinel cages. In our facility, one tablespoon of dirty bedding from each cage on a rack is added to a designated sentinel cage every 3 wk at cage change, and PCR testing is performed on in-cage filter paper quarterly. We hypothesized that cages that received Helicobacter spp.-positive bedding at later time points would have a lower detection rate of Helicobacter spp. with PCR testing compared with cages that received positive bedding at earlier time points due to the filter paper becoming saturated. To determine if screening would be able to detect one positive row of cages on a rack, 9 tablespoons of Helicobacter-positive bedding and 71 tablespoons of negative bedding were added at the 3-, 6-, or 9-wk time points to 14 empty sentinel cages per time point. Negative bedding was added every 3 wk to cages not scheduled to receive positive bedding. Negative controls received 80 tablespoons of negative bedding and positive controls received 80 tablespoons of positive bedding at each time point. Filter paper was tested via PCR for Helicobacter spp. at 12 wk. All positive controls tested positive, and all negative controls tested negative. Two 3-wk cages, two 6-wk cages, and three 9-wk cages were positive, indicating no difference between time points. This resulted in a 16.7% Helicobacter spp. detection rate. These results indicate that PCR on in-cage filter paper may not be reliable in detecting low levels of Helicobacter spp. nucleic acid in dirty bedding.
小鼠感染螺旋杆菌会对肠胃、生殖和免疫系统产生广泛的影响。这可能会给研究带来重大的混杂变量,降低科学的严谨性。筛选小鼠群落中的螺旋杆菌物种可通过在无动物的脏垫料哨兵笼中放置的滤纸上进行非侵入性 PCR 测试来实现。在我们的设施中,每 3 周换笼时都会从架子上的每个笼子中取一汤匙脏垫料放入指定的哨兵笼中,每季度在笼内滤纸上进行一次 PCR 检测。我们假设,与在较早时间点收到阳性垫料的笼子相比,在较晚时间点收到螺旋杆菌阳性垫料的笼子,由于滤纸已经饱和,PCR 检测的螺旋杆菌检出率会更低。为了确定筛查是否能够检测到笼架上的一排阳性笼子,在 3、6 或 9 周的时间点,在每个时间点的 14 个空哨兵笼中添加了 9 汤匙的螺旋杆菌阳性垫料和 71 汤匙的阴性垫料。阴性垫料每 3 周添加一次到未安排添加阳性垫料的笼子中。在每个时间点,阴性对照组添加 80 汤匙阴性垫料,阳性对照组添加 80 汤匙阳性垫料。12 周时,通过 PCR 对滤纸进行螺旋杆菌检测。所有阳性对照组检测结果均为阳性,所有阴性对照组检测结果均为阴性。两个 3 周的笼子、两个 6 周的笼子和三个 9 周的笼子都呈阳性,表明时间点之间没有差异。这导致 16.7% 的螺旋杆菌检出率。这些结果表明,用笼内滤纸进行 PCR 检测脏垫料中的低水平螺旋杆菌核酸可能并不可靠。
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Journal of the American Association for Laboratory Animal Science : JAALAS
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