Colony Of Rats Research Articles

Systematic analysis reveals that colony housing aligns gait profiles and strengthens link between histological and micro-CT bone markers in rat models of osteoarthritis.

Osteoarthritis (OA) etiopathogenesis is complex with strong environmental/lifestyle determinants that, in laboratory animals, extend to social context and stress levels. This study seeks to identify whether colony housing of rats exerts a social impact on locomotion behaviors to influence alignment between symptomatic (gait) and structural (bone micro-CT measures, cartilage morphometry, and histology) OA outcome measures. Rats were randomly allocated to conventional (type IV; n=48) or rat colony cage (RCC; n=30) housing, further randomized to OA surgical models (ACLT+tMx, MMT or DMM) or no surgery (control), and maintained for 19weeks during which multiple gait recordings were made. Standard histological grading and bone micro-CT data were collected at necropsy. Principal component analysis was used to summarize the variation in gait, micro-CT or histology. Linear mixed effects model or two-way ANOVA was employed to evaluate the impact of the housing system, surgery and time on gait, or micro-CT and histology components Analyses reveal that RCC exaggerates trends in gait change via a combined effect of the housing system and surgery. Intriguingly, RCC-housed nonoperated control rats showed similar gait changes to rats subjected to surgery; the latter exhibited significant structural joint changes in both systems. Stronger correlation between histological and micro-CT bone changes were found in medial and lateral tibia joint compartments of rats housed in RCC system. This study has established that rat social housing exaggerates outcomes in traditional histological measures of OA, generates stronger links between histology and micro-CT bone changes and removes gait differences as a variable in their etiology.

Demographic drivers of Norway rat populations from urban slums in Brazil

The Norway rat is a globally distributed pest, known for its resilience to eradication and control programs. Efficient population control, especially in urban settings, is dependent on knowledge of rat demography and population ecology. We analyzed the relationship between four demographic outcomes, estimated by live-trapping data, and fine-scale environmental features measured at the capture site. Wounds, a proxy for agonistic interactions, were associated with mature individuals. Areas with environmental features favorable to rats, such as open sewers and unpaved earth, were associated with more mature individuals with a better body condition index. The control measures (environmental stressors) are likely to be disrupting the social structure of rat colonies, increasing the frequency and distribution of agonistic interactions, which were common in both sexes and maturity states. The relationship between the favorable environmental conditions and the demographic markers analyzed indicate possible targets for infestation control through environmental manipulation, and could be incorporated into current pest management programs to achieve long-term success. Our study indicate that urban interventions focused on removal of potential resources for rats could be potential long-term solutions by reducing the carrying capacity of the environment.

Colony housing promotes structural and functional changes during surgically induced osteoarthritis in rats

The aim of the study was to investigate how social housing with high locomotion activity affects experimental osteoarthritis (OA) in rats. Rats were housed either conventionally in type IV cages in pairs or in rat colony cages (RCC) on 4 levels interconnected by jump holes or staircase in groups of 48. OA was induced by anterior cruciate ligament transection and resection of the medial meniscus (ACLT+tMx), medial meniscal tear (MMT) or destabilization of the medial meniscus (DMM). Functional changes were characterized by continues tracking of individual activity and catwalk gait analysis. Cartilage volume and bone structure were investigated at week 20 after surgery by histology and micro-CT. In the RCC, healthy rats changed cage levels 82±15 times daily, reduced by 30% after ACLT+tMx (p<0.0001). In both housing systems, the order of severity of the investigated models was ACLT+tMx>MMT>DMM in all outcome measures. Compared to Type IV, RCC housed rats developed stronger gait disturbance symptoms (ACLT+tMx; 95%CI=-15-2; p<0.004), the cartilage volume was reduced (ACLT+tMx: 95%CI=-0.1-0.5; p<0.0001), serum levels of the cartilage remodeling marker AGNx1 were higher (MMT; 95%CI=-53-(-6); p=0.001), bone was denser with increased volume (ACLT+tMx; 95%CI=0.8-7.5; p=0.004) and joints were less flexible (ACLT+tMx; 95%CI=3.6-14; p<0.0001). Housing rats in an environment allowing increased locomotion and socialization promotes structural and functional alterations during joint instability-induced OA. This increases the assay window, improves the relevance for the human disease and enables to discriminate the models in structural and behavioral parameters.

Investigation and Therapeutic Management of Hymenolepiosis in Wistar Rats from Chhattisgarh

The present investigation was aimed to know the cause of sudden mortality among Wistar rats in a rat colony. The dead rats (n=20) were subjected to necropsy examination. Intestinal lumens were examined to know the presence of parasites or parasitic body parts. Intestinal faecal contents were collected for identification of the parasitic eggs. Intestinal tissues were collected for histopathological examination. Necropsy revealed the presence of cestodes in the intestinal lumen. The intestines were found to be haemorrhagic and filled with serous and blood tinged contents. The cestode worms were identified as Hymenolepis diminuta based on gross morphological examination of the adult worms and microscopic examination of the eggs. Histopathology revealed necrosis of enterocytes, blunting of villi, degeneration and necrosis of villous and crypt epithelium. The diseased rats were observed for clinical signs and symptoms. Clinically, the rats showed inappetence, dullness, depression, loss of body weight, rough hair coat and a typical hunchback posture. The affected rats were treated by oral administration of Praziquantel, Pyrantel Pamoate and Febantel combination (kiwof plus tablet, Savavet) at a dose rate of 6 mg/kg body weight. The rats were cured after anthelmintic treatment. The present study highlighted the occurrence of hymenolepiosis that caused high mortality among Wistar rats. This study emphasizes the necessity of rapid and specific disease diagnosis and treatment to control the mortality among affected rats and to prevent and control the further spread of parasitic infection among susceptible rats and in the personnel in close surroundings of the infected rats.

Methods for Establishment and Maintenance of Germ-Free Rat Models.

Numerous studies have demonstrated that the gut microbiota plays a vital role in human health and disease development. Although the number of studies on host–microbiota interactions have increased in recent years, the underlying pathogenesis of dysbiosis-related diseases are still largely unknown. Germ-free (GF) rodent models, with the animals housed in sterile isolators and completely free of microbiota, are useful tools to advance our understanding of host–microbiota relationship in vivo. Although protocols concerning the establishment and maintenance of GF mouse models have previously been reported, the establishment, maintenance and monitoring of GF rodents are labor-intensive, tedious and take experience and skills. The aim of our study was to establish a GF rat model for the following microbiota-related researches and provide an easy-to-use protocol for the establishment and maintenance of GF rat model in detail, including steps to set up the isolator, sterilize the flexible isolator bubble, import food, water and other supplies, and methods to acquire newborn GF rats, hand rearing of suckling GF rats and reproduction of GF offspring. During the hand feeding period, the body weight of suckling GF rats was weighed once a day to ascertain the amount of artificial milk was given. Based on our results, the body weight of suckling GF rats decreased 1 week after birth and then began to increase. Methods for verifying the quality of the model like assessing the sterile status of the rat colony are also described. Moreover, possible difficulties and challenges, especially during gavage, and suggestions to avoid contamination will be discussed. The protocol presented will facilitate the establishment of GF rat models and downstream microbiota-related researches.

PCR SURVEILLANCE OF FOUR PROTOTYPIC RAT PARVOVIRUSES IN LABORATORY RAT COLONIES IN TAIWAN

Prototypic rat parvoviruses are among the most common infectious agents in laboratory rats worldwide. It is important to determine the infectious condition of rat parvoviruses in research colonies in Taiwan. In this study, virus species-specific PCR assays were applied to screen Kilham rat virus (KRV), Toolan’s H-1 virus (H-1 virus), rat parvovirus (RPV) and rat minute virus (RMV) in 203 rats from five different research colonies in Taiwan. In this survey, the infection rate of rat parvoviruses was 19.2% (39/203), including KRV (12.3%; 25/203), RMV (8.9%; 18/203) and RPV (1.0%; 2/203). Dual infection of KRV and RMV were detected in 3.0% (6/203) of samples. No rats were positive for H-1 virus. The infectious rates of KRV and RMV detected in this study are much higher than those reported in USA and in Europe. Rat parvovirus PCR assays reported in this paper could be a useful tool for routine rat health monitoring program applied in laboratory rat facilities. [Chang YH, Liao YC, Wang KW, Hsieh SC, Wang MH, Fang BH, Chang CY, Chueh LL, Wan CH, PCR Surveillance of four prototypic rat parvoviruses in laboratory rat colonies in Taiwan, Taiwan Vet J 46(1): 1–7, 2020]

Conventional and Specific-Pathogen Free Rats Respond Differently to Anesthesia and Surgical Trauma

Specific-pathogen free (SPF) animals were introduced in the 1960s to minimize disease and infection as variables in biomedical research. Our aim was to examine differences in physiological response in rat colonies bred and housed in a conventional versus SPF facility, and implications for research. Sprague-Dawley rats were anesthetized and catheterized for blood and pressure monitoring, and electrocardiogram (ECG) leads implanted. Hematology was assessed, and coagulation profile using rotational thromboelastometry. Health screening was outsourced to Cerberus Sciences. SPF rats had significantly lower pulse pressure (38% decrease), arrhythmias and prolonged QTc (27% increase) compared to conventional rats. No arrhythmias were found in conventional rats. SPF rats had significantly higher white cell, monocyte, neutrophil and lymphocyte counts, and were hyperfibrinolytic, indicated by EXTEM maximum lysis >15%. Independent assessment revealed similar pathogen exclusion between colonies, with the exception of Proteus in SPF animals. Returning to a conventional facility restored normal host physiology. We conclude that SPF animals displayed an abnormal hemodynamic, hematological and hemostatic phenotype in response to anesthesia and surgery, and provide a number of recommendations to help standardize research outcomes and translation.

To be or not to be pink(1): contradictory findings in an animal model for Parkinson's disease.

The PTEN-induced putative kinase 1 knockout rat (Pink1−/−) is marketed as an established model for Parkinson’s disease, characterized by development of motor deficits and progressive degeneration of half the dopaminergic neurons in the substantia nigra pars compacta by 8 months of age. In this study, we address our concerns about the reproducibility of the Pink1−/− rat model. We evaluated behavioural function, number of substantia nigra dopaminergic neurons and extracellular striatal dopamine concentrations by in vivo microdialysis. Strikingly, we and others failed to observe any loss of dopaminergic neurons in 8-month-old male Pink1−/− rats. To understand this variability, we compared key experimental parameters from the different studies and provide explanations for contradictory findings. Although Pink1−/− rats developed behavioural deficits, these could not be attributed to nigrostriatal degeneration as there was no loss of dopaminergic neurons in the substantia nigra and no changes in neurotransmitter levels in the striatum. To maximize the benefit of Parkinson’s disease research and limit the unnecessary use of laboratory animals, it is essential that the research community is aware of the limits of this animal model. Additional research is needed to identify reasons for inconsistency between Pink1−/− rat colonies and why degeneration in the substantia nigra is not consistent.

The haematological profile with fifty generations of undernutrition cannot be reversed with six generations of a recuperation diet in a wistar rat colony

The haematological profile with fifty generations of undernutrition cannot be reversed with six generations of a recuperation diet in a wistar rat colony

A semi-nested RT-PCR assay for detection of norovirus in rat fecal samples.

Norovirus is a highly prevalent pathogen that can infect a wide range of host species. Thus far, there have only been two reports of norovirus infection in rats. Diagnostic assays for the detection of norovirus are well established, but a specific molecular assay for the diagnosis of norovirus infection in laboratory rats has not yet been reported. In this study, we describe the development of a sensitive, semi-nested RT-PCR assay for detection of norovirus in fecal samples from Rattus norvegicus, reared in animal facilities under different sanitary barrier conditions. Additionally, we describe the first report of the presence of norovirus in rat colonies from Brazilian animal facilities.

Organ Generation from Knockedout Rat Blastocysts Complemented with Pluripotent Stem Cells.

Regeneration of human organs in domestic animal model would provide enough number of functional donor organs in transplantation therapy. Recent progresses in pluripotent stem cells and nuclease-based genome editing tools have set the stage for investigating the chimeric complementation approach to generate functional organs from embryonic stem (ES) cells or induced pluripotent stem (iPS) cells. In this chapter, protocol for allogeneic or xenogeneic organ generation using knocked-out (KO) rat blastocysts and the rat or mouse ES/iPS cells is described. The protocol includes (1) the preparation of KO rat colony, (2) the preparation of rat or mouse ES/iPS cells, (3) the recovery of rat blastocysts, (4) the stem cell injection into blastocysts, (5) the embryo transfer into pseudopregnant recipient uteri, and (6) the genotyping and organogenetic analysis of chimeric offspring. The accumulation of basic and practical knowledge in the rodent model would be useful in improving therapeutic performance to regenerate 3D organs available for transplantation.

Temporal and Space-Use Changes by Rats in Response to Predation by Feral Cats in an Urban Ecosystem

Feral cats (Felis catus) are predators that cause widespread loss of native wildlife in urban ecosystems. Despite these risks, cats are commonly released as control agents for city rats (Rattus spp.). Cats can influence their prey directly by killing or indirectly through changes to feeding or space-use. However, cats prefer defenseless prey, and there are no data suggesting that cats influence large (>300g) urban rats. We used a pre-existing radiofrequency identification assay (microchipped rats and field cameras) and ethograms to assess the impact of cats, including temporal and space use patterns, on an active rat colony. From Dec 27, 2017 through May 28, 2018 we captured 306 videos of pre-identified cats and/or rats that shared the same space. There were three instances of predation and 20 stalking events. Logistic regression showed the likelihood of a rat being seen on a particular day is associated with the number of cats seen on the same day (OR=0.1, p<0.001) or previous day (OR=0.15, p<0.001). Space-use was also impacted. For every additional cat sighting, a rat is 1.19 times more likely to move in the direction of shelter. Our findings of low levels of predation support why ecologists believe the risks to native wildlife outweighs any benefits of releasing cats. Even though rats were less likely to be seen, they simply shifted their movements and remained present in the system. Our findings that cat presence led to fewer rat sightings may explain the common perception of their value as rat-predators despite the associated risks.

Wild Norway Rats Do Not Avoid Predator Scents When Collecting Food in a Familiar Habitat: A Field Study

The ability to avoid predators is crucial to wild prey animals’ survival. Potential danger is signalled, among others, by the presence of predator scents. These odors are used in research both to trigger and to study fear reactions in laboratory animals; they are also employed as repellents against pest rodent species. In our study, we assessed nine predator-derived odors for their effectiveness in eliciting avoidance responses in a free-living colony of Norway rats (Rattus norvegicus). The rats were studied in a field setting. Food was put in two compartments inside the experimental pen: in one of them, predator scent was introduced on experimental days. The rats did not avoid boxes with predator odor and did not display an increased latency of food-carrying behavior or any other fear-related behavior, such as freezing or increased grooming. The results confirm the hypothesis that the foraging of rodents in a well-known territory and in relative proximity to burrows and other shelters is not affected by indirect cues of predation risk, such as the presence of predator urine or feces. We have also concluded that in a well-established colony living in a familiar territory, predator scent holds little promise as rodent repellent.

Genetic diversity and sex ratio of naked mole rat, Heterocephalus glaber, zoo populations.

The naked mole rat, Heterocephalus glaber, is a highly unusual mammal that displays a complex social system similar to that found in eusocial insects. Colonies of H. glaber are commonly maintained in zoo collections because they represent fascinating educational exhibits for the public. However, little is known about the genetic structure or sex ratio of captive populations of H. glaber. In this study, we developed a set of microsatellite markers to examine genetic variation in three captive zoo populations of H. glaber. We also studied sex ratio of these captive populations. Our goal was to determine levels of genetic variation within, and genetic differences between, captive populations of H. glaber. Overall, we found modest levels of genetic variation in zoo populations. We also uncovered little evidence for inbreeding within the captive populations. However, zoo populations did differ genetically, which may reflect the isolation of captive naked mole rat colonies. Finally, we found no evidence of biased sex ratios within colonies. Overall, our study documents levels of genetic variation and sex ratios in a captive eusocial mammalian population. Our results may provide insight into how to manage captive populations of H. glaber.

Rat polyomavirus 2 infection in a colony of X-linked severe combined immunodeficiency rats in Japan.

Polyomaviruses (PyVs) infect a wide range of animals and provoke wasting diseases, particularly in immunosuppressed hosts. Recently, a novel Rattus norvegicus polyomavirus 2 (RatPyV2) has been identified in a colony of X-linked severe combined immunodeficiency (X-SCID) rats in the United States. Here, we describe the first report of the RatPyV2 infection in an X-SCID rat colony in Japan. The affected rats exhibited adult-onset wasting. Histologically, we observed large basophilic intranuclear inclusion bodies within the hyperplastic or dysplastic epithelial cells in the salivary glands, Harderian glands, extraorbital lacrimal glands, and in respiratory and reproductive tissues. Among these organs, the parotid salivary, Harderian, and extraorbital lacrimal glands were most obviously affected. In particular, the parotid salivary glands were the most severely and diffusely affected and atrophic lesions were prominent even at 1 month of age, which suggested that the parotid salivary glands would be highly susceptible to RatPyV2 in X-SCID rats. RatPyV2 inclusion bodies were also detected in the tail of the epididymis and deferent duct. Such reproductive lesions developed significantly in the later stage of breeding age, and therefore may be associated with the reduced fecundity observed in the infected X-SCID rats. We also established a simple, rapid, and non-invasive diagnostic method based on the Amp-FTA method, using buccal swabs for the detection of RatPyV2 in immunodeficient rats. Our findings contribute to the early detection and diagnosis of RatPyV2 infections.

Spatial variation in the parasite communities and genomic structure of urban rats in New York City.

Brown rats (Rattus norvegicus) are a globally distributed pest. Urban habitats can support large infestations of rats, posing a potential risk to public health from the parasites and pathogens they carry. Despite the potential influence of rodent-borne zoonotic diseases on human health, it is unclear how urban habitats affect the structure and transmission dynamics of ectoparasite and microbial communities (all referred to as "parasites" hereafter) among rat colonies. In this study, we use ecological data on parasites and genomic sequencing of their rat hosts to examine associations between spatial proximity, genetic relatedness and the parasite communities associated with 133 rats at five sites in sections of New York City with persistent rat infestations. We build on previous work showing that rats in New York carry a wide variety of parasites and report that these communities differ significantly among sites, even across small geographical distances. Ectoparasite community similarity was positively associated with geographical proximity; however, there was no general association between distance and microbial communities of rats. Sites with greater overall parasite diversity also had rats with greater infection levels and parasite species richness. Parasite community similarity among sites was not linked to genetic relatedness of rats, suggesting that these communities are not associated with genetic similarity among host individuals or host dispersal among sites. Discriminant analysis identified site-specific associations of several parasite species, suggesting that the presence of some species within parasite communities may allow researchers to determine the sites of origin for newly sampled rats. The results of our study help clarify the roles that colony structure and geographical proximity play in determining the ecology of R.norvegicus as a significant urban reservoir of zoonotic diseases. Our study also highlights the spatial variation present in urban rat parasite communities, indicating that rats across New York City are not reservoirs for a homogenous set of parasites and pathogens. As a result, the epidemiological risks may be similarly heterogeneous for people in urban habitats.

Clinical, pathological, and molecular investigation of Mycoplasma pulmonis-induced murine respiratory mycoplasmosis in a rat (Rattus norvegicus) colony.

Aim:Mycoplasma pulmonis (MP) remains potentially important rodent pathogen causing murine respiratory mycoplasmosis (MRM) which may go undiagnosed due to its asymptomatic nature. In the present study, we carried out clinical, pathological, and molecular investigations of MP-induced MRM in a rat colony.Materials and Methods:Two female Wistar rats were observed to be diseased in animal facility of NISER, Bhubaneswar, and were kept in isolation for further investigation. Both the animals were found to be positive for MP after serological and molecular tests. Thereafter, whole rat colony comprising of 36 animals was segregated based on clinical symptoms and further sampled for histopathological, serological, and molecular investigations. Tracheal washing and infected lung tissue were collected during necropsy examination for DNA extraction. Molecular diagnosis was done by polymerase chain reaction (PCR) assay using species-specific primers.Result::Classical symptoms of MP-associated respiratory tract infection were observed in only 2 of 36 infected animals, and most of the animals were found asymptomatic to the disease; however, all the animals were found to be carrier after necropsy and PCR assay. Gross and histopathological finding suggested severe congestion of the lungs along with suppurative and necrotizing pneumonia. The disease is confirmed by molecular diagnosis using species-specific primers in PCR assay.Conclusion:MRM may go undiagnosed due to asymptomatic nature. Detailed study of clinical symptoms, pathology, serology, and PCR-based molecular approach may aid in health monitoring and detection of MRM in a rodent colony reared for experimental purpose.

High prevalence of Seoul hantavirus in a breeding colony of pet rats.

As part of further investigations into three linked haemorrhagic fever with renal syndrome (HFRS) cases in Wales and England, 21 rats from a breeding colony in Cherwell, and three rats from a household in Cheltenham were screened for hantavirus. Hantavirus RNA was detected in either the lungs and/or kidney of 17/21 (81%) of the Cherwell rats tested, higher than previously detected by blood testing alone (7/21, 33%), and in the kidneys of all three Cheltenham rats. The partial L gene sequences obtained from 10 of the Cherwell rats and the three Cheltenham rats were identical to each other and the previously reported UK Cherwell strain. Seoul hantavirus (SEOV) RNA was detected in the heart, kidney, lung, salivary gland and spleen (but not in the liver) of an individual rat from the Cherwell colony suspected of being the source of SEOV. Serum from 20/20 of the Cherwell rats and two associated HFRS cases had high levels of SEOV-specific antibodies (by virus neutralisation). The high prevalence of SEOV in both sites and the moderately severe disease in the pet rat owners suggest that SEOV in pet rats poses a greater public health risk than previously considered.

Polyomavirus-associated Prostatitis in Wistar Han Rats Following Immunosuppression in a Chronic Toxicity Study.

Chronic prostatitis characterized on light microscopic examination by moderate, multifocal, predominantly lymphocytic inflammation associated with epithelial atypia and intranuclear and cytoplasmic inclusion-like material was identified in the prostate gland of 2 Wistar Han rats administered an immunomodulatory test article in a 6-month chronic toxicity study. Transmission electron microscopy of the prostate glands identified 45-nm, nonenveloped, icosahedral virions arranged in paracrystalline array within the cell nuclei in 1 of the 2 rats. The size, shape, location, and array pattern were most consistent with a polyomavirus. The light and electron microscopic findings after immunosuppression in our case have a resemblance to a polyomavirus recently reported to affect prostate gland epithelium in a colony of immunocompromised X-linked severe combined immune deficiency rats. To the best of our knowledge, this is the first report of light and electronic microscopic lesions in the reproductive tract associated with polyomavirus following chronic immunosuppression in a widely used, wild-type Wistar Han rat.

Survival rates of homozygotic Tp53 knockout rats as a tool for preclinical assessment of cancer prevention and treatment

BackgroundThe gene that encodes tumor protein p53, Tp53, is mutated or silenced in most human cancers and is recognized as one of the most important cancer drivers. Homozygotic Tp53 knockout mice, which develop lethal cancers early in their lives, are already used in cancer prevention studies, and now Tp53 knockout rats have also been generated. This study assessed feasibility of using homozygous Tp53 knockout rats to evaluate the possible outcome of cancer chemoprevention.MethodsA small colony of Tp53 knockout rats with a Wistar strain genetic background was initiated and maintained in the animal house at our institution. Tp53 heterozygotic females were bred with Tp53 homozygous knockout males to obtain a surplus of knockout homozygotes. To evaluate the reproducibility of their lifespan, 4 groups of Tp53 homozygous knockout male rats born during consecutive quarters of the year were kept behind a sanitary barrier in a controlled environment until they reached a moribund state. Their individual lifespan data were used to construct quarterly survival curves.ResultsThe four consecutive quarterly survival curves were highly reproducible. They were combined into a single “master” curve for use as a reference in intervention studies. The average lifespan of untreated male Tp53 homozygous knockout rats was normally distributed, with a median of 133 days. Sample size vs. effect calculations revealed that confirming a 20% and 30% increase in the lifespan would respectively require a sample size of 18 and 9 animals (when assessed using the t-test with a power of 80% and alpha set at 0.05). As an example, the Tp53 homozygous knockout rat model was used to test the chemopreventive properties of carnosine, a dipeptide with suspected anticancer properties possibly involving modulation of the mTOR pathway. The result was negative.ConclusionFurther evaluation of the Tp53 homozygous knockout male rat colony is required before it can be confirmed as a viable tool for assessing new methods of cancer prevention or treatment.