Background Data for General Toxicology Parameters in RccHan:WIST Rats at 8, 10, 19 and 32 Weeks of Age

Recently, RccHan(TM):WIST (Wistar Hannover) rats were introduced to toxicity studies in Japan. The present study was performed to obtain control data for general toxicological parameters as an aid for interpretation of results in toxicity studies using this strain of rats. Four test groups comprising of 25 male and 25 female RccHan(TM):WIST rats were housed for 2, 4, 13 or 26 weeks from 6 weeks of age and observed and examined for clinical observation, body weight, food consumption, urinalysis, hematology, blood chemistry, organ weight, necropsy and/or histopathology. Ophthalmological examination was not conducted in this study, and the data in this report were obtained from an ongoing 104-week background study in RccHan(TM):WIST rats. These data were compared with the historical control data of CD(SD) (Sprague-Dawley) and/or F344 (Fischer) rats. The body weights of RccHan(TM):WIST rats were lower than those of CD(SD) rats and higher than those of F344 rats. The ophthalmological examination revealed a greater incidence of focal corneal opacity. Histopathology revealed focal mineralization of the cornea and Berlin blue-positive pigmentation in the epididymal interstitium as well as hepatocytes. Other than the above, some minor differences were found in urinalysis, hematology, blood chemistry and organ weights as compared with CD(SD) rats.     

Hematology and clinical chemistry values in pregnant Wistar Hannover rats compared with nonmated controls

BACKGROUND: The Wistar Hannover rat has been considered as an alternative animal model to the Sprague-Dawley rat in the safety evaluation of candidate pharmaceuticals for potential reproductive and developmental toxicity. Hematology and clinical chemistry results may provide useful evidence of maternal toxicity in the absence of fetal effects. OBJECTIVE: The purpose of this study was to evaluate differences in routine laboratory values between nonmated and pregnant (near-term)Wistar Hannover rats during a control developmental study. METHODS: One hundred fifty pregnant female Wistar Hannover rats (Tac:Glx:WlfBR) were dosed orally once per day with distilled water from gestation days (GDs) 6 through 17. An additional 150 nonmated (nonpregnant) females used as age-matched controls were dosed from study days (SDs) 7 through 18. Blood samples were collected on GD 18 or 19 (SD 19 or 20) for routine hematology and plasma clinical chemistry tests. Reference intervals were established for pregnant and nonmated animals. RESULTS: On GD 18/19, pregnant rats had a lower RBC count, hemoglobin concentration, and HCT, and higher MCH, MCHC, reticulocyte percentage, and platelet, WBC, absolute reticulocyte, segmented neutrophil, lymphocyte, and monocyte counts compared with nonmated rats. Pregnant rats had lower albumin, glucose, urea, and chloride concentrations, lower creatine kinase and alkaline phosphatase activities, higher total bilirubin, cholesterol, triglyceride, calcium, phosphorus, and globulin concentrations, and higher ALT activity than nonmated rats. Serum triglyceride concentration was approximately fourfold higher in pregnant rats compared with nonmated controls. CONCLUSION: Differences in hematology and chemistry values in pregnant Wistar Hannover rats are similar to those in Sprague-Dawley rats and support use of the Wistar Hannover rat as an animal model in the assessment of maternal toxicity. Differences in laboratory values of pregnant rats should be considered when interpreting data following exposure to candidate pharmaceuticals.     

Background Data on Developmental Parameters During the Gestation Period in Rats

Abstract: Background data during the gestation period were obtained from 128 Wistar Hannover GALAS rats and 26 Crl:CD(SD) pregnant rats in the control groups of our previous toxicity studies. The body weights of dams in the Wistar Hannover GALAS rats were significantly lower throughout the gestation period than those in the Crl:CD(SD) rats. In contrast, the time-dependent change in the body weight gain (%) of dams showed very similar trends in both strains. The mean number of live embryos/fetuses in the Wistar Hannover GALAS rats was 12.0, and was lower than that (14.5) in the Crl:CD(SD) rats. The placental weights gradually increased with pregnancy progression and reached a plateau on gestation day (GD) 19, although the embryo/fetal weights rapidly increased from GD 17 to GD 21. The embryo/fetal weights in the Wistar Hannover GALAS rats were significantly lower on only GD 21 than those in the Crl:CD(SD) rats. It is considered that this fetal weight difference between the strains develops during the fetal period, but not during the organogenesis period. In contrast, there were no differences in the placental weights between the two strains. Microscopically, the thickness of the labyrinth zone in the Wistar Hannover GALAS rats was thicker throughout the gestation period than that in the Crl:CD(SD) rats.     

Histological Characteristics of the Regression of Corpora Lutea in Wistar Hannover Rats: the Comparisons with Sprague-Dawley Rats

We examined the ovaries of 44 Wistar Hannover (RccHan^TM:WIST) (WH) and 30 Sprague-Dawley (SD) rats at 32-weeks of age to determine whether the ovarian structure and formation/regression of the corpora lutea (CLs) differ between the two strains. The average ovary weight was higher in WH rats. The average number of all CLs, including currently formed and previously formed CLs, was higher in WH rats in all cycles; however, no appreciable difference was detected in the number of newly or currently formed CLs between the two strains. CLs regression characterized by degeneration and necrosis of luteal cells began to appear in diestrus in both strains; however, the distribution of degenerated/necrotic cells in CLs differed. Necrotic cells were scattered in SD rats but were focally observed in the center of the CL in WH rats. The reduction in size of previously formed CLs accompanied by regression started about 2 or more stages later in WH rats than in those of SD rats. In conclusion, the higher number of CLs in WH rats is considered to be due to slow CL regression compared with in SD rats.     

Uterine Carcinosarcoma in a 2-year-old Female Wistar Hannover GALAS Rat

Carcinosarcomas are rare tumors in humans as well as rats and most commonly occur in the uterus. Recently, we observed a case of incidental carcinosarcoma of the uterus in a female Wistar Hannover GALAS [BrlHan:WIST@ Jcl (GALAS)] rat at 2 years of age. Histopathologically, the tumor was characterized by an admixture of malignant epithelial and nonepithelial elements. The carcinomatous components represented a type of endometrial carcinoma, consisting of glandular and solid proliferation of large-sized tumor cells. Prominent mitoses and tumor cell invasion were observed. The sarcomatous components were characterized by multifocal proliferation of severe atypical cells with cartilage matrix and were diagnosed as chondrosarcoma. Transitions between carcinomatous and sarcomatous components were observed, and many tumor cells in the solid lesion showed immunohistochemical reactivity with both cytokeratin and vimentin. Based on these findings, this tumor was diagnosed as a uterine carcinosarcoma. This is the first report of uterine carcinosarcoma in Wistar Hannover GALAS [BrlHan:WIST@Jcl (GALAS)] rats.     

链脲佐菌素诱导1型糖尿病大鼠模型的影响因素探讨

通过注射链脲佐菌素(STZ)制作大鼠1型糖尿病模型,观察大鼠品系、给药剂量、给药次数对大鼠成模率、死亡率的影响,同时研究利用口服葡萄糖耐量试验(OGTT)判断大鼠糖尿病成模率的意义。随机取16只SD大鼠设置正常对照组。试验一:Wistar大鼠随机分为中剂量组(55 mg/kg,n=12)和高剂量组(65 mg/kg,n=12);试验二:SD大鼠一次性腹腔注射STZ(55 mg/kg,n=12),与1中的Wistar大鼠作对比;试验三:SD大鼠随机分为一次给药组(n=16)和两次给药组(n=16),注射剂量均为55 mg/kg,观察期间进行OGTT。结果表明,Wistar大鼠成模率和死亡率均高于SD大鼠;采用SD大鼠、中剂量给药和两次给药的方式可提高成模率,并降低死亡率;在有明确胰岛病理改变的模型组大鼠,其OGTT异常阳性率显著高于空腹血糖异常阳性率。用STZ诱导糖尿病模型是一种稳定可靠的方法。     

Establishment of a Model of Spontaneously-Running-Tokushima-Shikoku Rats with Left Atrial Thrombosis

Studies that investigate the underlying mechanisms of disease and treatment options typically require the use of a suitable animal model. Few suitable animal models exist for left atrial thrombosis. Here, we demonstrated that the Spontaneously-Running-Tokushima-Shikoku (SPORTS) rat — a Wistar strain known for its running ability—is predisposed to the development of thrombi in the left atrium. We investigated the incidence of left atrial thrombosis in male (n = 16) and female (n = 17) SPORTS rats and observed organized atrial thrombosis in 57% and 38% of males and female rats, respectively. In the male rats, systolic blood pressures and heart rates were significantly higher in SPORTS rats than in control Wistar rats. We could not find any evidence of arrhythmias, such as atrial fibrillation, during electrocardiographic examination of SPORTS rats. We believe that the SPORTS rat could serve as a new research model for left atrial thrombosis; further, it may be suitable for research investigating the development of new antithrombotic approaches for the control of atrial thrombosis or familial thrombophilia in humans.     

不同比例虫草培养基混合料对SPF级Wistar大鼠血液生化指标的影响

为了探讨不同比例的虫草培养基混合料对SPF级Wistar大鼠血液生化指标的影响， 收集3组共60只Wistar大鼠的18项血液生化指标进行统计分析。结果发现，试验组Wistar大鼠的血红蛋白、嗜酸性细胞的水平均显著低于对照组；试验组雄性Wistar大鼠的胆固醇、甘油三酯水平均显著低于对照组，尿素氮和血糖显著高于对照组；试验组雌性Wistar大鼠的谷丙转氨酶和谷草转氨酶水平均显著高于对照组，而胆固醇和甘油三酯水平均显著低于对照组；其他指标各组间和不同性别间均无显著差异。研究结果表明，饲喂不同比例的虫草培养基混合料，可使SPF级Wistar大鼠的部分血液生化指标发生明显改变。      

Carcinogen-induced Thyroid Proliferative Lesions in Wistar Hannover GALAS Rats with Thyroid Dysplasia

Incidences and morphological features of thyroid proliferative lesions induced by carcinogens in Wistar Hannover GALAS rats (GALAS rats) showing normal growth with or without thyroid dysplasia were examined. All thyroid tissue samples were obtained from our recently conducted study using male GALAS rats treated with 5 carcinogens according to the medium-term multiorgan carcinogenicity bioassay protocol (called DMBDD treatment). In the DMBDD-treated rats, thyroid dysplasia was found in 9 out of 114 rats. Follicular cell adenomas were found in 5 out of 9 rats with thyroid dysplasia and in 7 out of 105 rats without thyroid dysplasia. The incidence of adenoma was significantly increased in rats with thyroid dysplasia (55.6%) compared with that in rats without thyroid dysplasia (6.7%). Adenomas in rats with thyroid dysplasia were observed as single or multiple nodules, well demarcated and composed of variously sized vacuolated cells or unvacuolated cells. These histopathological features and staining profiles of luminal colloid for PAS and thyroglobulin, together with PCNA-positive cells, were fundamentally similar to those of rats without thyroid dysplasia. On the other hand, the luminal colloid in adenomas of rats with thyroid dysplasia had a tendency to be poorly stained for T(4) compared with that of rats without thyroid dysplasia. From these findings, it appears that dysplastic thyroids of rats showing normal growth are more sensitive to carcinogens than normal thyroids. In addition, the morphological features of carcinogen-induced thyroid proliferative lesions in GALAS rats with thyroid dysplasia were fundamentally similar to those of rats without thyroid dysplasia, except for the vacuoles and T(4) staining profile.     

Pancreatic Ductal Adenocarcinoma in a Wistar Hannover GALAS Rat

There are no reported spontaneous cases of pancreatic ductal adenocarcinoma (PDAC), and there are few reports about chemically-induced PDAC in rats. We encountered a PDAC in a Wistar Hannover GALAS rat that had been subjected to a medium-term multiorgan carcinogenicity bioassay. This article describes the histological and histochemical findings of the tumor. The tumor was located in the pancreatic tissue and had not invaded the liver parenchyma or the mucosal layer of the alimentary tract. The tumor cells were atypical and were mainly arranged in small tubules. In addition, abundant stroma and mucus production were observed in the tumor. In an immunohistochemical examination, the tumor cells were positive for cytokeratin, Sox9 and pancreas duodenum homeobox 1 and negative for amylase 2A and insulin. Therefore, the tumor was diagnosed as a PDAC based on its histological and histochemical findings. We considered that the tumor was caused by the carcinogens administered during the abovementioned bioassay.     

Lesions in the Larynx of Wistar RccHan: WIST Rats

Specific regions in the rat larynx exhibit cellular changes in response to inhaled xenobiotics. These regions include the base of the epiglottis, ventral pouch, and medial surfaces of the vocal processes of the arytenoid cartilages. 1 , 2 In order to collect information on the usefulness of trimming techniques, the influence of different vehicles, the impact of different application routes in toxicity studies, and differences between induced vs. spontaneous lesions, the data obtained from a large number of inhalation and non-inhalation studies performed in Wistar RCCHan(TM): Wist rats at Harlan Laboratories Ltd Switzerland, all evaluated or reviewed by the same pathologist, were compiled for a detailed review. The value of different trimming techniques was deemed to be greatest for transverse and sagittolongitudinal section techniques, as compared to horizontolongitudinally section techniques. The comparison of lesions encountered in control rats of inhalation studies treated with different vehicles did not reveal differences in the type, distribution pattern, incidence and/or severity of spontaneous lesions. The types of lesions were also independent of different application routes in non-inhalation studies compared to inhalation studies. The pattern of spontaneous lesions in the rodent larynx was determined by degenerative and inflammatory lesions starting most often in the submucosal glands by desiccated secretion followed by mineralization and local inflammation or were induced by impacted foreign bodies. Squamous metaplasia was recorded in the respiratory epithelium overlaying the ventral gland as a spontaneous lesion in male Wistar rats from inhalation studies with a maxim of 20.0% in an inhalation oncogenicity study. Induced metaplastic changes recorded in the larynx were reversible. Other induced lesions in inhalation studies consisted of submucosal edema, necrosis, inflammation and/or granuloma. Induced lesions in non-inhalation studies were found to be exclusively related to reflux laryngitis or food impaction. It is concluded, that in rodents induced lesions of the larynx differ in type, distribution pattern, severity and incidence from spontaneous lesions.     

Mineralized corneal sequestrum in a cat.

An 8-month-old 3-kg (6.6-lb) sexually intact male cat was evaluated for chronic refractory keratitis and a corneal plaque that developed after treatment with a corticosteroid-containing ophthalmic preparation. Pertinent ophthalmic findings included blepharospasm, conjunctivitis, corneal vascularization, and a tan raised corneal plaque with a dense and gritty composition. Lamellar keratectomy was performed to excise the plaque. The cornea healed with mild scarring. Histologic examination revealed extensive coagulation necrosis and mineralization of the corneal stroma with increased inflammatory cells, blood vessels, and fibrosis, which is compatible with a diagnosis of mineralized corneal sequestrum. The history suggested chronic feline herpesvirus-1 (FHV-1)-induced keratoconjunctivitis as the underlying cause. Topical corticosteroid administration may have potentiated the preexisting corneal necrosis and initiated mineralization. To our knowledge, mineralization of a corneal sequestrum in a cat has not been reported in a clinical case but has been reported in cats experimentally infected with FHV-1.     

芦丁对大鼠免疫器官指数的影响

研究天然植物提取物芦丁对大鼠免疫器官指数的影响,考察芦丁调节动物免疫功能的作用.选择16只Wistar雌性青春期大鼠,随机分为2组,每组8只,分别为对照组(每日每只灌胃生理盐水2 mL)、芦丁组(每日灌服芦丁60 mg/kg体重),正式试验为期2周.12只Wistar受孕母鼠,随机分为2组,每组6只,相同处理,从哺乳第...     

蝗虫多肽对大鼠肥胖抑制作用的研究

试验旨在研究能量型日粮中添加蝗虫多肽对大鼠肥胖及脂肪代谢的影响。试验选用30只6周龄的Wistar- Imamichi系雄性大鼠,随机分为3组,试验1组在基础日粮中添加20%猪油;试验2组在基础日粮中添加20%猪油+添加10%蝗虫多肽粉末;试验3组在基础日粮中添加20%猪油+添加10%纤维素粉,试验期8个月。试验期间每周末称重一次,试验结束后测定血液生化指标及各脏器中瘦素(Leptin)浓度,并运用Real-time PCR技术对脂肪中瘦素mRNA表达量进行了测定。结果表明,试验2组与试验1、3组相比,从18周龄开始体重增加明显减缓,对附睾周围脂肪重量存在极显著的抑制(P＜0.01),各脏器重量未因使用蝗虫多肽而变化;部分血液生化指标存在显著或极显著差异(P＜0.05,P＜0.01);试验2、3组的血清、脑脊髓液中瘦素浓度极显著低于试验1组(P＜0.01);试验2组瘦素mRNA表达量显著高于试验1组(P＜0.05)。说明蝗虫多肽所含多种功能性酶能够抑制大鼠体内脂肪的增加,有效地控制了肥胖。     

Factors affecting premature chromosome condensation of cumulus cell nuclei injected into rat oocytes

To date, production of cloned rats by somatic cell nuclear transfer (NT) has not yet been successful. Inducing premature chromosome condensation (PCC) of injected cell nuclei in recipient cytoplasm is considered essential for successful mouse cloning by the Honolulu method. In the present study, some factors affecting PCC of rat cumulus cell nuclei injected into rat oocytes were examined. Wistar female rats (young: 4 to 5-week-old, mature: > or =10-week-old) were superovulated by injections of eCG and hCG, and oocytes recovered 14 or 17 h after hCG injection were received with cumulus cell nuclei using piezo-driven micromanipulator. When the oocytes were recovered 14 h post-hCG injection from young rats and the nuclear injection into oocytes was completed within 45 min, PCC was observed in 44-49% of NT oocytes. In the case of oocytes from mature rats, PCC occurred in 11-19% of the NT oocytes. Oocytes recovered 17 h post-hCG injection did not support PCC of the injected nuclei (0-7%) regardless of the donor age. Treatment of oocytes with a neutral cysteine protease inhibitor, N-acetylleucylleucylnorleucinal, slightly increased the incidence of PCC (48 vs 37%). Comparison of rat strains for oocyte donors indicated that proportions of NT oocytes undergoing PCC in Wistar and LEW oocytes (41-46%) were higher than those in Donryu and F344 oocytes (17-25%). Thus, ability of rat oocytes to promote PCC of the injected nuclei is dependent on the characteristics of oocytes, such as age or strain of donor rats, and timing of oocyte recovery.     

Strain differences in morphometrical characteristics of rat kidneys

To clarify the strain differences in the morphology of the rat kidneys, we investigated the morphometrical characteristics of the kidneys of Slc:Wistar, Slc:SD, and F344/NSlc rats. The diameter of the renal corpuscles in female F344/N rats is smaller than that in female Wistar rats. Although sex differences (males>females) were shown in SD and F344/N rats, no effects of castration were detected in any of the groups. Strain-dependent differences in the percentage of renal corpuscles with a cuboidal parietal layer were found in both male and female groups. The highest percentage of them was noted in male Wistar rats. Effects of castration were observed in female Wistar and male F344/N rats, and the values after castration were significantly higher than those in the intact animals. As for the number of proximal convoluted tubular nuclei, no strain differences were detected in either the male or female groups. Although a sex difference was found in SD rats (female>male), no effects of castration were detected in any of the groups. In female F344/N rats, numerous numbers of PAS-positive granules, which were observed in the proximal convoluted and straight tubular epithelia, were noted. Orchiectomy induced an increase of these granules in male SD and F344/N rats, but ovariectomy showed no effects on them in any strains. This is the first study to clarify the strain differences in the morphological characteristics of the kidneys in ordinary rat strains.     

Availability of a Microglia and Macrophage Marker,Iba-1, for Differential Diagnosis of Spontaneous Malignant Reticuloses from Astrocytomas in Rats

In rats, it is sometimes difficult to distinguish malignant reticuloses from astrocytomas in routine histopathological assessment. In the present study, four spontaneous brain neoplasms developing in the cerebrum of one Wistar Hannover rat and three Sprague-Dawley rats were immunohistochemically examined using microglia and macrophage markers. Histopathologically, these neoplasms were localized mainly in the cerebral cortex, hypothalamus or piriform lobe, and the portions showing solid growth did not show characteristic cellular arrangement but had an indistinct boundary with the surrounding brain parenchyma. Neoplastic cells had oval or pleomorphic small nuclei with abundant eosinophilic cytoplasm. Two cases showed neoplastic cell infiltration into the meninges and perivascular spaces. Silver staining showed lack of reticulin fiber production in the stroma of the neoplasms. Immunohistochemically, the neoplastic cells were strongly positive for Iba-1 and sporadically positive for CD68 in all four cases. On the basis of these results, all the neoplasms examined here could be distinguished from astrocytomas and diagnosed as malignant reticuloses. Thus, immunohistochemical demonstration of microglia/macrophage characters, such as using Iba-1, is considered to be helpful for differential diagnosis of malignant reticuloses from astrocytomas among spontaneously occurring primary brain neoplasms in rats.     

Corneal sensitivity in healthy, immature, and adult alpacas

Objective To determine the corneal sensitivity of healthy, immature, and adult alpacas to establish normal reference values for this species. Animals studied Six normal crias and 18 normal adult alpacas. Procedures Corneal sensitivity was determined by evaluating the corneal touch threshold (CTT) in five regions of the cornea using a Cochet–Bonnet aesthesiometer. The nylon filament length in cm was then converted to applied pressure values in mg/0.0113 mm² and g/mm² using a conversion chart. Results The central region of the cornea was the most sensitive, and the dorsal and temporal regions were the least sensitive in alpacas. There were no significant differences between the right and left eyes or between values in males and females at any site. Crias had significantly greater corneal sensitivity compared with adult alpacas in all five regions of the cornea that were evaluated. Conclusions Normal reference values for CTT in alpacas were established using a Cochet–Bonnet aesthesiometer. The central region of the cornea is the most sensitive in alpacas, and crias have higher corneal sensitivity than adult alpacas.     

Meniscal Mineralization in Domestic Cats

Objective: To (1) determine prevalence of radiographically detectable meniscal mineralization in domestic cats and (2) to evaluate the association between meniscal mineralization and degenerative joint disease (DJD). Study Design: Prospective study. Animals: Client‐owned cats (n=100) and 30 feline cadavers. Methods: Randomly selected client‐owned cats were used to determine the prevalence of meniscal mineralization. Stifles from feline cadavers were used to evaluate the relationship between meniscal mineralization (using high‐resolution X‐ray), radiographic DJD, and cartilage damage. Menisci were evaluated histologically. Results: Forty‐six percent of the client‐owned cats had meniscal mineralization detected in 1 or both stifles. Pain scores were not significantly different between stifles with meniscal mineralization and those with no radiographic pathology (P=.38). Thirty‐four of 57 cadaver stifles had meniscal mineralization, which was always located in the cranial horn of the medial meniscus. Percentage mineralization of the menisci was significantly correlated with the cartilage damage score of the medial femoral (r²=0.6; P<.0001) and tibial (r²=0.5; P<.0001) condyles as well as with the total joint cartilage damage (r²=0.36; P<.0001) score and DJD score (r²=0.8; P<.0001). Conclusion: Meniscal mineralization is a common condition in domestic cats and seems to indicate medial compartment DJD. Clinical Relevance: Clinical significance of meniscal mineralization is uncertain. Further work is needed to determine if the meniscal mineralization is a cause, or a consequence of joint degeneration.     

大鼠肺鳞癌模型的建立

探讨和构建Wistar大鼠肺癌的动物模型,以了解肺癌发生发展的过程.采用Wistar大鼠68只,其中试验组60只大鼠(A组30只:三甲基胆蒽(MCA) 5 mg～7 mg,二乙基亚硝胺(DEN) 0.02 mL;B组30只:MCA 7.5 mg～9.5 mg,DEN 0.03 mL),左肺下叶支气管灌注含MCA和DEN的碘油溶液0.1 mL;另8只灌注0.1 mL碘油,作为对照组.试验的3月和6月末分别进行X线检测,发现3月末已有肺部阴影,6月末有50%出现明显的肺部阴影.组织学观察发现1月末已出现肺部原位癌和早期浸润癌;6月末48只发生癌变,致癌率为80%,癌变的Wistar大鼠肺标本中有多个阶段病变共存;共获取支气管黏膜上皮增生44例,不典型增生29例(含鳞状化生7例),原位癌18例,侵袭癌19例,转移癌11例.结果表明,一次性支气管灌注MCA和DEN可引起大鼠肺鳞癌的发生,并获得癌发生过程中各个阶段的标本,该疾病模型为人类肺癌发生机制的研究提供了基础依据.