Breeding and rearing of Japanese field voles (Microtus montebelli Milne-Edwards) and Hungarian voles (Microtus arvalis Pallas) as new herbivorous laboratory animal species

Microtus species are very useful as experimental small animals with herbivorous characteristics. In this report, to approach establishment of the use of Japanese field voles (M. montebelli) and Hungarian voles (M. arvalis) as experimental animals, the biological characteristics, breeding methods and reproductive performance of these animal species were comparatively investigated. The number of chromosomes of M. arvalis is 12n = 46, differing from those of M. montebelli, 2n = 30. The breeding and reproduction of the Microtus species were relatively easily accomplished in the laboratory, by giving high fiber-containing rations. These animals prove to be polyestrous and showed postpartum estrous on the day of parturition. In the breeding by the continuous mating of the same pair, there was little or no delay in implantation due to lactation, especially in M. arvalis resulting in continuous birth at intervals of 20 to 21 days. On examination of vaginal smears, Japanese field voles (M. montebelli) did not show any definite pattern whereas 23 out of 30 voles from Hungary (M. arvalis) examined showed 6- to 18-day cycles. There were remarkable differences between the Japanese voles and the Hungarian voles, both biologically and reproductively.     

Laboratory breeding of the red-backed vole (Clethrionomys rufocanus bedfordiae)]

Laboratory matings were attempted to establish breeding colonies of red-backed voles (Clethrionomys rufocanus bedformidae) as experimental animals. For these mating, 10 pairs of red-backed voles which were captured in the Tohbetsu region of Hokkaido, Japan and their litters were used. In the results for two years, 1987 to 1988, the rates of pregnancy, birth and weaning were 35.4%, 94.5% and 79.5%, respectively. The mean litter size was 5:1 +/- 1.6 with a range of 1 to 9. The mean gestation period was 20.0 +/- 0.7 days with a range of 18 to 22. These results suggest that planned production of red-backed voles in the laboratory is possible. To determine intraregional variations of red-backed voles with a view to the establishment of a strain by inbreeding, restriction patterns of mitochondrial DNAs using seven restriction endonucleases were compared. Four different patterns were obtained from wild red-backed voles used in the present study.     

NMR spectroscopy based metabonomic studies on the comparative biochemistry of the kidney and urine of the bank vole (Clethrionomys glareolus), wood mouse (Apodemus sylvaticus), white toothed shrew (Crocidura suaveolens) and the laboratory rat

The metabolic profiles of three wild mammals that vary in their trophic strategies, the herbivorous bank vole (Clethrionomys glareolus), the granivorous wood mouse (Apodemus sylvaticus), and the insectivorous white-toothed shrew (Crocidura suaveolens), were compared with that of a widely used strain of laboratory rat (Sprague Dawley). In conjunction with NMR spectroscopic investigations into the urine and blood plasma composition for these mammals, high resolution magic angle spinning (HRMAS) 1H-nuclear magnetic resonance (NMR) spectroscopy was applied to investigate the composition of intact kidney samples. Adaptation to natural diet affects both renal metabolism and urinary profiles, and while these techniques have been used to study the metabolism of the laboratory rat little is known about wild small mammals. The species were readily separated by their urinary profiles using either crude metabolite ratios or statistical pattern recognition. Bank vole urine contained higher concentrations of aromatic amino acids compared with the other small mammals, while the laboratory rats produced relatively more hippurate. HRMAS 1H-NMR demonstrated striking differences in both lipid concentration and composition between the wild mammals and Sprague Dawley rats. Bank voles contained high concentrations of the aromatic amino acids phenylalanine, tyrosine and tryptophan in all tissue and biofluids studied. This study demonstrates the analytical power of combined NMR techniques for the study of inter-species metabolism and further demonstrates that metabolic data acquired on laboratory animals cannot be extended to wild species.     

Impact of season on seminal characteristics and endocrine status of adult free-ranging African buffalo (Syncerus caffer)

Pituitary, gonadal and adrenal activity were compared in free-living, adult African buffalo bulls during the breeding and nonbreeding seasons. Frequent blood samples were collected for 2 h from anaesthetized bulls treated intravenously with saline, gonadotrophin-releasing hormone (GnRH, 200 micrograms), human chorionic gonadotrophin (hCG, 10,000 i.u.) or adrenocorticotrophic hormone (ACTH, 1.5 mg). Electroejaculates also were collected from anaesthetized bulls during the breeding and nonbreeding seasons. Pretreatment testosterone concentrations among bulls varied more during the breeding (0.17-23.0 ng/ml) than the nonbreeding (0.15-2.21 ng/ml) season. The variation within the breeding season was attributed to 8 of 25 bulls producing higher (P less than 0.05) serum testosterone (High-T; 16.28 +/- 2.03 ng/ml) and testicular LH receptor (1.53 +/- 0.22 fmol/mg testis) concentrations compared with their seasonal counterparts (Low-T; 0.95 +/- 0.26 ng/ml; 0.38 +/- 0.04 fmol/mg) or with all bulls during the nonbreeding season (0.90 +/- 0.27 ng/ml; 0.31 +/- 0.04 fmol/mg). The magnitude of GnRH- and hCG-induced increases in serum testosterone was similar (P greater than 0.05) between Low-T bulls and bulls during the nonbreeding season. In the High-T animals treated with GnRH or hCG, serum testosterone did not increase, suggesting that secretion was already maximal. Peak serum LH concentrations after GnRH were greater (P less than 0.05) in bulls during the nonbreeding than the breeding season; FSH responses were similar (P greater than 0.05). ACTH treatment did not increase serum cortisol concentrations above the 2-fold increase measured in bulls treated with saline, hCG and GnRH (P greater than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)     

High spatial resolution measurements of organ blood flow in small laboratory animals

With the use of a newly developed Imaging Cryomicrotome to determine the spatial location of fluorescent microspheres in organs, we validate and report our processing algorithms for measuring regional blood flow in small laboratory animals. Microspheres (15-microm diameter) of four different fluorescent colors and one radioactive label were simultaneously injected into the left ventricle of a pig. The heart and kidneys were dissected, and the numbers of fluorescent and radioactive microspheres were determined in 10 randomly selected pieces. All microsphere counts fell well within the 95% expected confidence limits as determined from the radioactive counts. Fluorescent microspheres (15-microm diameter) of four different colors were also injected into the tail vein of a rat and the left ventricle of a rabbit. After correction for Poisson noise, correlation coefficients between the colors were 0.99 +/- 0.02 (means +/- SD) for the rabbit heart and 0.99 +/- 0.02 for the rat lung. Mathematical dissection algorithms, statistics to analyze the spatial data, and methods to visualize blood flow distributions in small animal organs are presented.     

Serum concentrations of gonadotropins and steroid hormones of Neophocaena phocaenoides asiaeorientalis in middle and lower regions of the Yangtze River

As a relatively isolated and unique freshwater population, the Yangtze finless porpoise (Neophocaena phocaenoides asiaeorientalis) is the most endangered subpopulation of this species. The objective of this study was to improve our understanding of their reproductive endocrinology by measuring (radioimmunoassay) serum concentrations of FSH, LH, estradiol (E(2)), progesterone (P(4)) and testosterone (T(2)) in free-ranging animals. Blood samples were collected from 66 Yangtze finless porpoises (41 males and 25 females) captured in the middle and lower reaches of the Yangtze River. Based on significant variation of serum T(2) concentrations in males with body length (BL) >138 cm, we inferred that they were mature; in these animals, there were significant seasonal variations in serum T(2) concentrations, with the highest concentrations in March and April (502.0+/-319.8 ng/dL, mean+/-S.D.) and the lowest in December (79.4+/-83.2 ng/dL). Serum T(2) concentrations positively correlated with serum concentrations of LH and weakly correlated with serum concentrations of E(2), whereas there was a significant negative correlation between serum LH and FSH concentrations in males >138 cm. The smallest apparently pregnant female porpoise had a BL of 130 cm. Serum P(4) concentrations ranged from 13.2 to 112.4 ng/mL (43.9+/-28.3 ng/mL) in pregnant females, and fluctuated under 1.0 ng/mL in non-pregnant females with BL>130 cm. Serum LH concentrations were significantly higher in non-pregnant females >130 cm versus those females <130 cm. To our knowledge, this is the first study of endocrine-related maturity and seasonal breeding characteristics of the Yangtze finless porpoise.     

银狐气味对根田鼠繁殖和觅食的影响

在实验室条件下, 主要测定银狐的气味对根田鼠社会行为、觅食、繁殖特征及性激素水平的影响, 并探讨银狐气味使根田鼠产生繁殖迟滞的机制。结果表明, 银狐气味对82 %的配对根田鼠产生显著的繁殖迟滞, 对根田鼠一般动作、探究、自我修饰及婚配行为的频次具有显著的抑制作用( P < 0105) 。对各种行为累积时间的作用则产生明显的性别差异, 银狐气味可提高雄体及雌体一般动作及雌体亲密行为的累积时间; 对雄体自我修饰行为, 以及雌体攻击、探究及自我修饰行为的累积时间有显著的降低作用( P < 0105) 。雄体附睾精子计数、雌体卵巢指数及胚胎重量极显著地减低( P < 0101) , 相应地, 雌体血清孕酮显著地提高( P< 0101) 。     

Reproductive features of the Russian vole in laboratory breeding.

Reproductive features of newly bred Russian voles (Microtus rossiaemeridionalis) as a laboratory animal were studied. This species is a copulatory ovulator, and most couples copulated 6 to 16 h after pairing. The gestation period varied from 18 to 22 days (mean +/- SD: 20.6 +/- 0.9, n = 72), and the average litter size was 4.6 +/- 1.9 (n = 125). Compared with the litter size at the first parturition (3.6 +/- 1.6, n = 72), the litter size in the subsequent parturitions increased to 5.9 +/- 1.4 (n = 53). The animals exhibited postpartum estrus, and repeated pregnancy accompanied with suckling pups and parturition continuously in the laboratory condition unlike other vole species. In view of their complex stomach and good proliferation, the Russian voles were evaluated as a good laboratory animal, especially as a model animal for ruminant studies.     

捕食风险对越冬根田鼠肠道寄生物易感性的影响

在自然生态系统中,不同营养级物种可通过特征介导间接效应对生态系统的稳定及种群产生深刻的影响。但目前有关特征介导间接效应的实验研究多见于无脊椎动物、鱼类和两爬类。本研究以根田鼠为对象,在野外围栏内建立预防捕食者和未预防捕食者两种实验处理种群,并通过采用麦克马斯特法测定两种处理种群实验个体肠道寄生物感染种类及感染率和感染强度,采用PHA（phytohemagglutinin）反应和白细胞分类计数测定不同处理种群实验个体免疫能力,以分析捕食风险对根田鼠肠道寄生物的感染效应。结果表明,未预防捕食者处理组根田鼠PHA反应、白细胞计数和淋巴细胞计数较预防捕食者处理组实验个体显著降低,而球虫 E. wenrichi 的感染率和感染强度则显著增加,但绦虫和线虫以及其他3种球虫的感染率和感染强度无显著差异。结果表明,捕食者可通过介导猎物免疫力特征而间接影响猎物肠道寄生物的感染,验证了本项提出的捕食风险可通过降低根田鼠的免疫能力而增加其肠道寄生物感染的假设。     

Comparison of male reproductive parameters in three rat strains: Dark Agouti, Sprague-Dawley and Wistar

The choice of experimental animal can have a large impact on experimental results, an example is the anecdotal evidence suggesting that Dark Agouti (DA) rats have a lower reproductive capacity than other rat strains. In this paper we report on an investigation into male reproductive characteristics in three rat strains--Wistar, Sprague-Dawley (outbred strains) and DA (an inbred strain). Reproductive organ weights, blood testosterone levels and sperm counts were measured in mature age-matched male rats. DA animals had significantly smaller testis weights than the Sprague-Dawley and Wistar animals, and this did not appear to be related to the overall smaller body mass of the DAs. There were no differences between the three strains in testicular histology or sperm counts (per gram testis). Although there was also no significant difference in epididymal sperm count, the DA animals had a much greater variability in sperm count than the other strains. There were no differences in relative (to body weight) epididymal, seminal vesicle or ventral prostate weights or in the blood testosterone levels. These results suggest that differences in reproductive capacity in DAs are neither the result of morphological differences in the reproductive organs nor in circulating testosterone levels. Sperm production appears to be normal but the lowered testicular weight and variability in epididymal sperm counts suggests that there are other factors in the testicular or epididymal environment which alter male reproductive function.     

Comparative aspects of microsomal cytochrome P-450-dependent monooxygenase activities in musk shrew (Suncus murinus), Mongolian gerbil (Meriones unguiculatus), harvest mouse (Micromys minutus) and rat.

1. The cytochrome P-450 content (0.75 +/- 0.13 nmol/mg microsomal protein) in musk shrew (suncus, Suncus murinus) liver microsomes was lower than that (1.30 +/- 0.26) in rat liver microsomes, but it is approximately the same level as in the Mongolian gerbil (Meriones unguiculatus, 1.18 +/- 0.14), harvest mouse (Micromys minutus, 1.11 +/- 0.02) and rat. 2. The hydroxylation activity (based on cytochrome P-450) of medium-chain fatty acids (otanoic, decanoic, lauric and tridecanoic acids) is much higher in suncus, Mongolian gerbil and harvest mouse than in rat, with the exception of the activity of decanoic and tridecanoic acids in Mongolian gerbil. 3. This suggests that cytochrome P-450 species catalyzing the hydroxylation of medium-chain fatty acids are present in these laboratory animals in higher concentrations. 4. The aminopyrine N-demethylation activity based on microsomal protein or cytochrome P-450 in suncus is significantly lower than that in rat, but the N-demethylation activity in Mongolian gerbil and harvest mouse is approximately 1.7-2.0-fold greater than that in rat.     

Successive changes in the blood composition of experimental normal beagle dogs associated with age

Eight hematologic parameter values, 16 serum biochemical constituents, serum protein fractions and albumin-globulin ratios were determined in blood samples obtained from 879 normal, healthy Beagle dogs of both sexes which had been reproduced and bred in our laboratories. The blood samples were collected from the Beagles that ranged in monthly ages from 1 to 12 and in monthly ages from 13 to 121, which were classified as the adult class. As a result, red blood cell counts, hemoglobin concentrations and packed cell volumes increased with growth. Red blood cell parameters of normal Beagles in our laboratories were rather higher than those in literatures presented by many other researchers. MCV decreased and MCHC increased gradually with age. Total serum protein concentrations increased with growth. alpha 1-1 and alpha 1-2 Globulin fractions descended, but beta 2 and gamma globulin fractions ascended in serum proteins. Alkaline phosphatase activities, inorganic phosphorus concentrations and glucose concentrations decreased conspicuously with growth. Leucine aminopeptidase activities and calcium concentrations decreased slightly. Serum cholinesterase and LDH activities showed a tendency to diminish similarly. Blood urea nitrogen and creatinine concentrations multiplied gradually. Hematologic parameters became almost steady in our 7-month-old dogs or older ones and serum biochemical constituents had a tendency to be stable in our 7- to 9-month-old dogs or older ones in the blood. White blood cell counts, alkaline phosphatase activities, inorganic phosphorus concentrations, glucose concentrations, leucine aminopeptidase activities and calcium concentrations were lowest in the adult class.     

Hematological and serum biochemical effects of nursing on the mother in cynomolgus monkeys (Macaca fascicularis)]

The effects of nursing on maternal hematological and serum biochemical values were analyzed in cynomolgus monkeys reared in indoor cages at Tsukuba Primate Center. In our breeding system, infants are usually separated from their mothers at the age of 121 to 180 days. Mother monkeys of such infants were studied hematologically and biochemically (Group B), as were mother monkeys who happened to have nursed their infants past 181 days after parturition (Group A). During the period with their infants, mother monkeys in the latter group showed lower white blood cell counts (WBC) and higher red blood cell counts (RBC), hematocrit values (Ht) and blood urea nitrogen concentrations (BUN) than the mother monkeys who had been separated from their infants. Also, serum calcium concentrations (Ca) were decreased with prolonged nursing periods, indicating that lactation by the mother monkey probably continues even for a period from 181 days to about one year after parturition if she nurses her infant. Lactation during this period may accelerate hematogenesis and protein metabolism in the mother monkey.     

The dynamics of health in wild field vole populations: a haematological perspective

1. Pathogens have been proposed as potentially important drivers of population dynamics, but while a few studies have investigated the impact of specific pathogens, the wealth of information provided by general indices of health has hardly been exploited. By evaluating haematological parameters in wild populations, our knowledge of the dynamics of health and infection may be better understood. 2. Here, haematological dynamics in natural populations of field voles are investigated to determine environmental and host factors associated with indicators of inflammatory response (counts of monocytes and neutrophils) and of condition: measures of immunological investment (lymphocyte counts) and aerobic capacity (red blood cell counts). 3. Individuals from three field vole populations were sampled monthly for 2 years. Comparisons with individuals kept under controlled conditions facilitated interpretation of field data. Mixed effects models were developed for each cell type to evaluate separately the effects of various factors on post-juvenile voles and mature breeding females. 4. There were three well-characterized 'physiological' seasons. The immunological investment appeared lowest in winter (lowest lymphocyte counts), but red blood cells were at their highest levels and indices of inflammatory response at their lowest. Spring was characterized by a fall in red blood cell counts and peaks in indicators of inflammatory response. During the course of summer-autumn, red blood cell counts recovered, the immunological investment increased and the indicators of inflammatory response decreased. 5. Poor body condition appeared to affect the inflammatory response (lower neutrophil and monocyte peaks) and the immunological investment (lower lymphocyte counts), providing evidence that the capacity to fight infection is dependent upon host condition. 6. Breeding early in the year was most likely in females in better condition (high lymphocyte and red blood cell counts). 7. All the haematological parameters were affected adversely by high population densities.     

Are degraded habitats from agricultural crops associated with elevated faecal glucocorticoids in a wild population of common vole (Microtus arvalis)?

The severe impact of agriculture on species’ abundance and diversity is widely recognized. However, its effects on the physiology of wild animal populations are poorly known. We analyzed faecal glucocorticoids levels in wild common voles (Microtus arvalis) living in a farmland landscape to test whether living in degraded habitats, such as crops, is correlated with increased glucocorticoids. Other factors such as sex, reproductive status, and population density were also considered. We captured voles with Sherman traps in crops and in their field margins which were comprised of semi-natural vegetation. We collected fresh faecal samples from captured individuals and quantified their levels of faecal corticosterone metabolites (FCM) in the laboratory. The quantification of FCM concentrations was performed by competitive enzyme immunoassay. Individuals captured within the crops had higher levels of FCM than those in field margins; females and breeding individuals exhibited higher FCM levels. In addition, FCM concentrations positively correlated with abundance of voles. Our results suggest that degraded habitats in agricultural landscapes are associated with increased glucocorticoid levels on common voles likely caused by a higher disturbance from agricultural practices and a lesser vegetation cover in crops compared with field margins.     

Hematologic values of normal Bolivian squirrel monkeys (Saimiri sciureus): a comparison between wild-caught and laboratory-bred male animals

Complete and differential blood counts were conducted on 23 male squirrel monkeys (Saimiri sciureus) of Bolivian origin. The sample included 9 laboratory-bred and 14 wild-caught adult monkeys weighing between 600 and 1,500 g. The means of total white blood cell (WBC) counts, total hemoglobin and hematocrits of laboratory-bred animals were found to be significantly different from those of wild-caught monkeys (p less than 0.01). Other hematologic parameters showed no significant differences between the two experimental groups. 3 of 14 (21.0%) wild-caught animals each had 1 reactive lymphocyte and 7% of the same group had 10 nucleated erythrocytes per 100 WBC. No reactive lymphocytes or nucleated erythrocytes were observed in laboratory-bred animals. The importance of these baseline data and the basis for the differences observed between laboratory and wild-caught monkeys are discussed within the context of experimental studies involving primates in which hematologic parameters are valuable.     

A quantitative study on recovery from acute haemorrhage in adult laboratory rats. I. Blood morphology

We evaluated changes in blood morphology for detecting acute blood loss in laboratory rats. Haematologic indices which are normally used in investigating drug effects on rat blood morphology were examined. Sublethal acute blood loss (25 ml per kg body weight) modified rat blood morphology in a few minutes; the blood picture was normal within two weeks. Blood drawing (usually up to 10 ml per kg b.w.) can modify rat blood morphology within one week. Erythrocyte counts, haemoglobin concentration and haematocrit decreased earliest, reticulocyte counts were changed longest. Comparing our results and literature data the recovery of blood morphology from acute haemorrhage seems to be more rapid in rats than in men.     

Breeding suppression between two unrelated and initially unfamiliar females occurs with or without social tolerance in common voles (<Emphasis Type="Italic">Microtus arvalis</Emphasis>)

Female common voles can breed in small groups or in isolation. Given the option, will isolated females opt for communal breeding with unrelated females and a probable low reproductive bias, or will they remain isolated, forgoing the advantages of group living? This laboratory work examined the response of two unrelated females to a foreign male in order to determine their social and breeding strategies. Before encountering a male, 70% of the females lived communally and 30% were solitary with a dominance hierarchy. In the presence of the male, only 33% of the females were still associated and lived with the male in a communal nest. In the other triads, only the oldest female lived with the male and she dominated the younger female. Although all animals were then experimentally separated to avoid late abortion due to social stress or infanticide, in 89% of the dyads only one female littered. This breeding suppression happened in hierarchic dyads but also in tolerant ones. This laboratory study on the social influence on reproduction showed that breeding suppression can occur in unrelated female common voles even when they are not closely grouped. It suggests that cooperative breeding between unrelated females should remain rare.     

野生来源TW小鼠近交系培育及其生物学特性的研究

目的　培育中国特有TW (TianjinWild)野生小鼠来源的实验近交小鼠品系 ,筛选其重要的生物学特征 ,丰富现有实验动物基因库。方法　严格按照国际培育近交系动物标准 ,即每代繁育的雌雄小鼠严格按照同胎雌雄 (兄妹 )进行交配。结果　经过 1 2年的精心培育 ,培育TW小鼠至F2 2代 ,其他 1 5地野生小鼠品系均因故中断。结论　该项研究将为丰富国际现有实验小鼠基因库资源 ,建立抗肺肿瘤特性实验小鼠模型动物奠定坚实基础。     

Effect of testosterone on n-methyl-d, l-aspartate-induced growth hormone secretion in male swine

Previous research from our laboratory demonstrated that n-methyl-d, l-aspartate (NMA), a potent agonist of glutamate, increased growth hormone (GH) secretion in barrows and boars. To determine if testosterone modulates NMA-induced GH secretion, Poland China x Yorkshire swine were challenged with NMA in a model that compared GH responses in boars with those of barrows or barrows treated with testosterone propionate (TP). Boars and barrows weighing 112.6+/-1.4 kg (mean +/- SE) were fitted with indwelling jugular vein catheters. Barrows (n = 16) were given i.m. injections of TP (25 mg in corn oil) twice daily from d 0 to d 6. Boars (n = 16) and control barrows (n = 15) received twice daily injections of corn oil. On d 6, blood was sampled every 15 min for 4 h. Two h after sampling began, all animals received an i.v. injection of NMA at a dose of 2.5 mg/kg body weight. Mean testosterone concentrations (ng/ml serum) were similar (P > .1) for boars (8.1+/-0.8) and barrows receiving TP (7.3+/-0.3), but were greater in both cases (P < .05) than for barrows receiving corn oil (.2+/-.01). Prior to NMA injections, mean GH concentrations were similar (P > .1) among groups and averaged 2.7+/-.2 ng/ml serum across treatments. Serum concentrations of GH after NMA increased (P < .05) similarly among groups and averaged 6.3+/-0.3 ng/ml across treatments during the 2-h period after injection. These results were not supportive of a role for testosterone as a modulator of NMA-induced GH secretion in male swine.