Experiences with the identification of small rodents

Own experiences with the identification of mice and rats using ear tags, ear punching, toe clipping (for newborns only) and ear tattooing are given and compared with methods presented in literature. By far, ear tattooing by the newly developed pincers is the most suitable method for marking adult rodents for life. Tattooing is easy to apply, easy to read and not harmful to the animals. For newborn pups on day 4 p. p., the s. c. injection of India ink into the palm of paw is recommended.     

Lurking in the shadows: emerging rodent infectious diseases

Rodent parvoviruses, Helicobacter spp., murine norovirus, and several other previously unknown infectious agents have emerged in laboratory rodents relatively recently. These agents have been discovered serendipitously or through active investigation of atypical serology results, cell culture contamination, unexpected histopathology, or previously unrecognized clinical disease syndromes. The potential research impact of these agents is not fully known. Infected rodents have demonstrated immunomodulation, tumor suppression, clinical disease (particularly in immunodeficient rodents), and histopathology. Perturbations of organismal and cellular physiology also likely occur. These agents posed unique challenges to laboratory animal resource programs once discovered; it was necessary to develop specific diagnostic assays and an understanding of their epidemiology and transmission routes before attempting eradication, and then evaluate eradication methods for efficacy. Even then management approaches varied significantly, from apathy to total exclusion, and such inconsistency has hindered the sharing and transfer of rodents among institutions, particularly for genetically modified rodent models that may not be readily available. As additional infectious agents are discovered in laboratory rodents in coming years, much of what researchers have learned from experiences with the recently identified pathogens will be applicable. This article provides an overview of the discovery, detection, and research impact of infectious agents recently identified in laboratory rodents. We also discuss emerging syndromes for which there is a suspected infectious etiology, and the unique challenges of managing newly emerging infectious agents.     

嗜肺巴斯德杆菌研究进展

嗜肺巴斯德杆菌(Pasteurella pneumotropica)是一种条件致病菌,人兽共患。主要危害啮齿类动物,特别是免疫缺陷或抑制的动物,可引起炎症和脓肿等症状。该菌是实验动物中感染率最高的病原菌之一,多呈隐性感染,给动物实验带来了极大的干扰。本文针对嗜肺巴斯德杆菌的流行病学、检测和鉴定、分子分型和防治等方面进行综述。     

Functional morphology of the ear in fossorial rodents, Microtus arvalis and Arvicola terrestris

Functionally relevant features and parameters of the outer, middle, and inner ear were studied morphologically and morphometrically in two species of voles, smaller Microtus arvalis and larger Arvicola terrestris. The findings in these fossorial (i.e., burrowing) rodents with components of surface activity were compared with respective findings reported for taxonomically related muroid rodents representing the same size classes but different eco-morphotypes: obligate subterranean rodents (Ellobius talpinus and Spalax ehrenbergi superspecies) and generalized rodents (Mus domesticus and Rattus norvegicus). The ear in voles was characterized by traits reported for subterranean rodents. The eardrum was round, without a distinct pars flaccida, and had an area of 5.4 mm2 in M. arvalis and 9 mm2 in A. terrestris. The middle ear exhibited reduced goniale, enlarged incus nearly parallel to the manubrium of the malleus. The malleus-incus lever ratio amounted to 2.1 (M. arvalis) and 2.0 (A. terrestris). The malleus-incus complex weighed about 0.8 mg in both vole species. The stapedial footplate had an area of 0.3 mm2 in M. arvalis and 0.4 mm2 in A. terrestris. The cochlea had 2.3 coils in both vole species; the basilar membrane was 8.5 mm and 10.5 mm long in M. arvalis and A. terrestris, respectively. There were on average 1,030 (M. arvalis) and 1,220 (A. terrestris) inner hair cells, and 3,760 (M. arvalis) and 4,250 (A. terrestris) outer hair cells in the organ of Corti. In quantitative terms, all these (as well as some further) traits and parameters were intermediate (related to body size) between those reported for generalized rodents on the one hand and subterranean ones on the other. The sound transmission system of the ear seems to be best tuned to frequencies of about 8-16 kHz with a high-frequency cut-off at about 50-60 kHz. The ear of A. terrestris seems to be tuned to somewhat lower frequencies than that in M. arvalis. In this aspect as well as regarding hearing sensitivity (as judged from the mechanical transmission parameters), voles can be considered intermediate not only in their lifestyle but also in their hearing abilities between the subterranean rodents (mole-vole and blind mole-rat) and the surface dwellers (house mouse and Norway rat).     

Identification by 16S rDNA fragment amplification and determination of genetic diversity by random amplified polymorphic DNA analysis of Pasteurella pneumotropica isolated from laboratory rodents

Pasteurella pneumotropica is an opportunistic bacterium frequently isolated from colonies of various laboratory rodents. Identification of this species, including its differentiation into two distinct biotypes (Jawetz and Heyl), is usually based on the use of conventional bacteriologic methods. In this study, a 16S rDNA fragment amplification procedure was developed for use as an alternative method for identification and differentiation of P. pneumotropica. Polymerase chain reaction (PCR) products were two distinctive fragments of 937 and 564 bp specific for biotypes Jawetz and Heyl, respectively. Specificity of PCR products could be achieved by EcoRI cleavage, leading to 596 plus 341-bp and 346 plus 218-bp fragments for each of the amplification products. Use of this procedure confirmed identification of 34 field isolates and allowed definitive identification of some strains that could not have been done by use of bacteriologic examinations. Field isolates subjected to random amplified polymorphic DNA (RAPD) analysis had high genetic diversity among biotype Jawetz strains in contrast to biotype Heyl strains. In conclusion, RAPD could represent an additional means for identification of ambiguous strains of biotype Heyl and a valuable epidemiologic tool for identification of biotype Jawetz strains of P. pneumotropica.     

Genomic DNA analysis from mouse toe lysates

The protocol described in this report provides a simple, accurate and efficient assay for detection of transgenes and mutations in large colonies of rodents, using crude lysates prepared from the digit cut from the animals for identification purposes. This can be done as early as 6 days of age, minimizing trauma to the mice and allowing assays to be completed long before weaning. Editor's note: The US Food and Drug Administration (FDA) had amended its regulations on good laboratory practice (GLP) for nonclinical laboratory studies regarding humane procedures for animal identification and expreimentation (Vol. 54 # 75). While discouraging use of toe clipping for animal identification, the FDA charged each facility's Institutional Animal Care and Use Committee (IACUC) with the responsibility for evaluation and final approval. C.A.P.     

模式小鼠总DNA三种提取方法比较

目的：建立简便、快捷、经济的模式小鼠总DNA提取方法,以快速鉴定大批量模式小鼠基因型。方法采用苯酚抽提法、异丙醇沉淀法、鼠耳煮沸法提取同种模式小鼠总DNA,对比DNA纯度、得率、耗费时间,并比较基因型鉴定结果。结果苯酚抽提法得率最高,异丙醇沉淀法最低;而纯度则按照苯酚抽提法、异丙醇沉淀法、鼠耳煮沸法顺序递减;在耗时上鼠耳煮沸法最短。三种方法提取的DNA均可做模版用于基因型鉴定。结论鼠耳煮沸法操作简单、成本最低,快速、基因型鉴定结果可靠,可用于规模化的基因型鉴定实验中。     

Diet,nesting density,and breeding success of rough-legged buzzards (Buteo lagopus) on the Nenetsky Ridge,Arctic Russia

The rough-legged buzzard, a circumpolar avian predator, is usually defined as rodent specialist in the tundra but as a generalist in the boreal zone, leaving open the question of where the shift in feeding strategy occurs. Here, we investigated the diet and breeding biology of buzzards as well as abundance of their possible prey during 5 years in the low-Arctic shrub tundra on the Nenetsky Ridge, Russia. We employed three complementary methods to assess the diet of this Arctic predator—pellet dissection, identification of prey remains on nests, and stable isotope analysis—to overcome their respective limitations. We documented fluctuations in abundances of the likely prey, namely rodents, ptarmigans, and hares. Nesting density of buzzards changed substantially over the years, but did not track the abundance cycle of the rodents. The number of buzzard fledglings was relatively low (1.08 ± 0.3) and did not change according to the density of rodents. In the year when rodents were at their lowest abundance, diet analyses of nestlings documented a shift from rodents to alternative prey, with a decrease in the proportion of tundra voles and an increase in proportion of hares, ptarmigans, and ducks. Here, we argue that buzzards may adopt different feeding strategies along the gradient from generalists to specialists. While the rough-legged buzzard is usually considered a small rodent specialist, our study shows that it can shift to alternative prey where or when rodents are scarce and when alternative prey are sufficiently abundant to provide subsistence for breeding.     

北京市大鼠、小鼠、豚鼠的病理学调查

目的：实验动物的健康状况对科学研究的发展有重要的影响,为了保证实验用鼠的健康质量,自从2009年起本实验室对北京市多家实验动物单位的实验用鼠进行了病理学抽检,从病理学的角度来调查北京市部分实验鼠的主要脏器的情况。方法我们主要对实验鼠的心、肝、脾、肺、肾、大肠和小肠进行取材,通过甲醛固定,石蜡切片HE染色、冰冻切片油红“O”染色、PAS染色等方法对实验用鼠进行病理学分析。结果根据病理学的调查结果,部分实验动物在以肝脏和肾脏为代表的器官中出现了不同程度的病变。结论结合对实验鼠的微生物学检测,分析认为,北京市的啮齿类实验动物中几乎无细菌、病毒和寄生虫等病原微生物的阳性感染个体,对人、动物及周围环境无明显危害;但是少部分实验鼠在临床上出现一定程度的病理学异常。     

被动式电子标签用于花鼠种群动态研究的可行性

在野生动物研究过程中经常需要对动物个体进行标记。传统的标记方法存在着标记物易脱落、可能影响被标记动物的生理和行为、难于进行个体识别等缺点。被动式电子标签(passive integrated transponder, PIT)是基于射频识别技术的电子标签,可用于体内标记,具有不易脱落消失且能够进行个体识别的优点。我们在凉水国家级自然保护区以PIT标签为标记物,采用标志重捕技术开展花鼠(Eutamias sibiricus)的种群动态研究,同时对应用PIT标签的可靠性进行了研究。2011年秋季至2012年春季共标记花鼠个体140只,其中86只被重捕至少1次。2011年秋季标记的72只个体中有38只在2012年春季被重捕。重捕的花鼠个体注射口愈合完好,体重无明显降低。结果表明PIT标签是一种安全可靠、识别迅速、数据准确、不易丢失、可循环使用的体内标记电子标签,适用于基于个体识别的小型哺乳动物生态学研究。     

Hantaviruses: an overview

Hantaviruses are a newly emerging group of rodent-borne viruses that have significant zoonotic potential. Human infection by hantaviruses can result in profound morbidity and mortality, with death rates as high as 50%, and potentially long-term cardiovascular consequences. Hantaviruses are carried by peridomestic and wild rodents worldwide and have occasionally been linked to infections in laboratory rodents. Because these viruses have been associated with significant human disease, they have become the subject of intense scientific investigation. In this review the reader is introduced to the hantaviruses, including hantavirus diseases and their pathogenesis. A review of the biology, morphology, and molecular biology of the hantaviruses with a brief overview of the ecology and biology of hantavirus-rodent pairs is also included. The risks of occupational exposure to hantaviruses, diagnosis of hantavirus infections, and methods for handling potentially infected rodents and tissues are discussed as well.     

Environmental enrichment for laboratory rodents: animal welfare and the methods of science

Because of the difficulty of' establishing objective measures of laboratory rodents' psychological well-being, developing environmental enrichment programs that are actually beneficial to rodents destined to participate in laboratory research is particularly challenging. Many studies of effects of environmental complexity, social housing, and increases in cage size suggest that professional judgments as to the impact of diverse types of environmental enrichment on rodent welfare are not a reliable basis for evaluating the outcomes of enrichment programs for laboratory rodents. Successful enrichment programs will vary from one rodent species to another, between sexes, as well as between age classes. There is a need for objective, measurable goals for proposed environmental enrichment programs for rodents, as well a s for empirical investigations of the beneficial and detrimental consequences of proposed environmental manipulations.     

Environmental Enrichment for Laboratory Rodents: Animal Welfare and the Methods of Science

Because of the difficulty of' establishing objective measures of laboratory rodents' psychological well-being, developing environmental enrichment programs that are actually beneficial to rodents destined to participate in laboratory research is particularly challenging. Many studies of effects of environmental complexity, social housing, and increases in cage size suggest that professional judgments as to the impact of diverse types of environmental enrichment on rodent welfare are not a reliable basis for evaluating the outcomes of enrichment programs for laboratory rodents. Successful enrichment programs will vary from one rodent species to another, between sexes, as well as between age classes. There is a need for objective, measurable goals for proposed environmental enrichment programs for rodents, as well a s for empirical investigations of the beneficial and detrimental consequences of proposed environmental manipulations.     

Does caloric restriction extend life in wild mice?

To investigate whether mice genetically unaltered by many generations of laboratory selection exhibit similar hormonal and demographic responses to caloric restriction (CR) as laboratory rodents, we performed CR on cohorts of genetically heterogeneous male mice which were grandoffspring of wild-caught ancestors. Although hormonal changes, specifically an increase in corticosterone and decrease in testosterone, mimicked those seen in laboratory-adapted rodents, we found no difference in mean longevity between ad libitum (AL) and CR dietary groups, although a maximum likelihood fitted Gompertz mortality model indicated a significantly shallower slope and higher intercept for the CR group. This result was due to higher mortality in CR animals early in life, but lower mortality late in life. A subset of animals may have exhibited the standard demographic response to CR in that the longest-lived 8.1% of our animals were all from the CR group. Despite the lack of a robust mean longevity difference between groups, we did note a strong anticancer effect of CR as seen in laboratory rodents. Three plausible interpretations of our results are the following: (1) animals not selected under laboratory conditions do not show the typical CR effect; (2) because wild-derived animals eat less when fed AL, our restriction regime was too severe to see the CR effect; or (3) there is genetic variation for the CR effect in wild populations; variants that respond to CR with extended life are inadvertently selected for under conditions of laboratory domestication.     

The presence of a correlation between erythrocyte acetylcholinesterase activity and body mass in small rodents

Studies have been made on the activity of erythrocyte acetylcholinesterase in 18 species of wild and laboratory rodents. Significant differences in the enzymic activity unrelated to taxonomic position of rodents were revealed. Negative correlation was found between the enzymic activity and body mass in rodents. The relationship between these parameters for wild rodents is approximated by an allometric equation Y = 1.1 . X-0.73, for laboratory species--Y = 1.5 . X-0.41. It is suggested that the revealed relationship indicates the existence of a connection between the activity of erythrocyte acetylcholinesterase and the intensity of metabolism in small rodents.     

Contributions of Electric and Acoustic Hearing to Bimodal Speech and Music Perception

Cochlear implant (CI) users have difficulty understanding speech in noisy listening conditions and perceiving music. Aided residual acoustic hearing in the contralateral ear can mitigate these limitations. The present study examined contributions of electric and acoustic hearing to speech understanding in noise and melodic pitch perception. Data was collected with the CI only, the hearing aid (HA) only, and both devices together (CI+HA). Speech reception thresholds (SRTs) were adaptively measured for simple sentences in speech babble. Melodic contour identification (MCI) was measured with and without a masker instrument; the fundamental frequency of the masker was varied to be overlapping or non-overlapping with the target contour. Results showed that the CI contributes primarily to bimodal speech perception and that the HA contributes primarily to bimodal melodic pitch perception. In general, CI+HA performance was slightly improved relative to the better ear alone (CI-only) for SRTs but not for MCI, with some subjects experiencing a decrease in bimodal MCI performance relative to the better ear alone (HA-only). Individual performance was highly variable, and the contribution of either device to bimodal perception was both subject- and task-dependent. The results suggest that individualized mapping of CIs and HAs may further improve bimodal speech and music perception.     

A Bead-Based Multiplex Assay for the Detection of DNA Viruses Infecting Laboratory Rodents

The Federation of European Laboratory Animal Science Association (FELASA) recommends screening of laboratory rodents and biological materials for a broad variety of bacterial agents, viruses, and parasites. Methods commonly used to date for pathogen detection are neither cost-effective nor time- and animal-efficient or uniform. However, an infection even if silent alters experimental results through changing the animals’ physiology and increases inter-individual variability. As a consequence higher numbers of animals and experiments are needed for valid and significant results. We developed a novel high-throughput multiplex assay, called rodent DNA virus finder (rDVF) for the simultaneous identification of 24 DNA viruses infecting mice and rats. We detected all 24 DNA viruses with high specificity and reproducibility. Detection limits for the different DNA viruses varied between 10 and 1000 copies per PCR. The validation of rDVF was done with DNA isolated from homogenised organs amplified by pathogen specific primers in one multiplex PCR. The biotinylated amplicons were detected via hybridisation to specific oligonucleotide probes coupled to spectrally distinct sets of fluorescent Luminex beads. In conclusion, rDVF may have the potential to replace conventional testing and may simplify and improve routine detection of DNA viruses infecting rodents.     

Ecological and Control Techniques for Sand Flies (Diptera: Psychodidae) Associated with Rodent Reservoirs of Leishmaniasis

Background

Leishmaniasis remains a global health problem because of the substantial holes that remain in our understanding of sand fly ecology and the failure of traditional vector control methods. The specific larval food source is unknown for all but a few sand fly species, and this is particularly true for the vectors of Leishmania parasites. We provide methods and materials that could be used to understand, and ultimately break, the transmission cycle of zoonotic cutaneous leishmaniasis.

Methods and Findings

We demonstrated in laboratory studies that analysis of the stable carbon and nitrogen isotopes found naturally in plant and animal tissues was highly effective for linking adult sand flies with their larval diet, without having to locate or capture the sand fly larvae themselves. In a field trial, we also demonstrated using this technique that half of captured adult sand flies had fed as larvae on rodent feces. Through the identification of rodent feces as a sand fly larval habitat, we now know that rodent baits containing insecticides that have been shown in previous studies to pass into the rodents'' feces and kill sand fly larvae also could play a future role in sand fly control. In a second study we showed that rubidium incorporated into rodent baits could be used to demonstrate the level of bloodfeeding by sand flies on baited rodents, and that the elimination of sand flies that feed on rodents can be achieved using baits containing an insecticide that circulates in the blood of baited rodents.

Conclusions

Combined, the techniques described could help to identify larval food sources of other important vectors of the protozoa that cause visceral or dermal leishmaniasis. Unveiling aspects of the life cycles of sand flies that could be targeted with insecticides would guide future sand fly control programs for prevention of leishmaniasis.     

A desert octodontid rodent,Tympanoctomys barrerae,uses modified hairs for stripping epidermal tissue from leaves of halophytic plants

Desert rodents that consume halophytic plants must have adaptations for coping with the high salt content of the leaves. A kidney capable of excreting very concentrated urine is one method. Another is removal of the hypersaline epidermis by means of chisel-like incisors prior to ingestion of the leaves. Tympanoctomys barrerae has evolved a unique refinement of the latter adaptation. It possesses two bundles of stiffened hairs on either side of the palate just caudal to the incisors. The bundles vibrate against the lower incisors, removing the epidermis from the leaves. The efficiency of the operation is significantly greater than with the use of incisors alone. Such a device has not been described in any other mammal. The facial muscles associated with the lips, the cheek vibrissae, and the oral cavity are described in T. barrerae and the nonhalophilic octodontid Octomys mimax. M. buccinatorius pars intermaxillaris is the only muscle in direct contact with the bristle bundles. Other anatomical features found in T. barrerae that may be associated with this feeding device are: 1) a much enlarged and mobile lower labial pad operated by Mm. buccinatorius pars orbicularis oris, pars longitudinalis profunda, and mandibularis cranialis profunda; 2) two oral glands not described in other rodents; and 3) a shortened tongue. Although, taken as a whole, this epidermal stripping device is unique to T. barrerae, most of its features have evolved by modification of structures present in the facial region of more generalized rodents.     

A primer on rodent identification methods

Identifying laboratory rodents as a group may be sufficient if all the animals are to receive the same treatment and/or manipulation and if individual variations in response are not to be recorded separately. However, it is frequently necessary to be able to differentiate between individual animals used in a study. Identification of individual animals may also be necessary to maintain health records and to properly manage colonies of laboratory animals in compliance with regulatory agencies and research protocol requirements.