The cotton rat (Sigmodon hispidus) is a permissive small animal model of human metapneumovirus infection, pathogenesis, and protective immunity

Human metapneumovirus (hMPV) is a newly described paramyxovirus that is an important cause of acute respiratory tract disease. We undertook to develop a small animal model of hMPV infection, pathogenesis, and protection. Hamsters, guinea pigs, cotton rats, and nine inbred strains of mice were inoculated intranasally with hMPV. The animals were sacrificed, and nasal and lung tissue virus yields were determined by plaque titration. None of the animals exhibited respiratory symptoms. The quantity of virus present in the nasal tissue ranged from 4.6 x 10(2) PFU/gram tissue (C3H mice) to greater than 10(5) PFU/gram (hamster). The amount of virus in the lungs was considerably less than in nasal tissue in each species tested, ranging from undetectable (<5 PFU/g; guinea pigs) to 1.8 x 10(5) PFU/gram (cotton rat). The peak virus titer in cotton rat lungs occurred on day 4 postinfection. hMPV-infected cotton rat lungs examined on day 4 postinfection exhibited histopathological changes consisting of peribronchial inflammatory infiltrates. Immunohistochemical staining detected virus only at the luminal surfaces of respiratory epithelial cells throughout the respiratory tract. hMPV-infected cotton rats mounted virus-neutralizing antibody responses and were partially protected against virus shedding and lung pathology on subsequent rechallenge with hMPV. Viral antigen was undetectable in the lungs on challenge of previously infected animals. This study demonstrates that the cotton rat is a permissive small animal model of hMPV infection that exhibits lung histopathology associated with infection and that primary infection protected animals against subsequent infection. This model will allow further in vivo studies of hMPV pathogenesis and evaluation of vaccine candidates.     

Pathogenesis of CAR bacillus in rabbits, guinea pigs, Syrian hamsters, and mice.

Cilia-associated respiratory (CAR) bacillus isolated from infected mice (designated, CBM) and propagated in embryonated chicken eggs was inoculated intranasally in rabbits (Oryctolagus cuniculus), guinea pigs (Cavia porcellus), hamsters (Mesocricetus auratus) and mice (Mus musculus). Gross and microscopic lesions, localization of CBM antigen in the respiratory tract, development of antibody, and ability to reisolate the CAR bacillus were studied in animals killed at 2-, 4-, or 8-week intervals postinoculation (PI). In rabbits, although no histopathological changes were observed in the respiratory tract, CBM antigen was detected on the ciliated epithelium of the respiratory tract, and serum CBM antibody was also detected 4 and 8 weeks PI. In guinea pigs, no histopathological changes were noted, CBM antigen was detected in the respiratory tract 2 and 4 weeks PI but not 8 weeks PI, and serum CBM antibody was detected 4 and 8 weeks PI. In hamsters, mononuclear cell proliferation in the submucosa of the bronchus and trachea was observed 8 weeks PI. CBM antigen was detected at first in the nasal cavity 2 weeks PI and in the lower respiratory tract 4 and 8 weeks PI and serum CBM antibody was detected 4 and 8 weeks PI. In mice, histopathological changes, CBM antigen and CBM antibody were observed. CBM was reisolated from the tracheal washouts of hamsters and mice 8 weeks PI but not from those of rabbits and guinea pigs. These results confirm and extend previous reports of experimentally-induced CAR bacillus infection in mice, guinea pigs, and rabbits. To this list of susceptible laboratory animals, we now add hamsters.(ABSTRACT TRUNCATED AT 250 WORDS)     

Vertebrate safety of Clostridium bifermentans serovar malaysia, a new larvicidal agent for vector control.

The safety of bacterial cells of Clostridium bifermentans serovar malaysia, which is highly toxic to mosquito larvae, was tested on mice, guinea pigs, rabbits, and goldfish. Inoculations of at least 1 x 10(8) cells per animal by routes recommended by World Health Organization (subcutaneous, percutaneous, inhalation, force-feeding, intraperitoneal, intravenous) and tests of subacute toxicity, anaphylactic shock, persistence in heart blood, and virulence by successive passages, were performed on mice, guinea pigs, or both. Growth was monitored for 1 mo before necropsy. Ocular irritation and skin scarification were tested with rabbits. C. bifermentans serovar malaysia did not induce any mortality or abnormal reactions in mammals at a dose 1,000 times higher than the level established by W.H.O. for the demonstration of safety. Bacterial cells are not toxic to goldfish at a dose 1,000 times higher than the LD50 for the target-mosquito larvae. We conclude that C. bifermentans serovar malaysia bacterial cells are safe for laboratory mammals and goldfish.     

Comparative studies of measuring condition of activated partial thromboplastin time and contact factors in experimental animals

For establishing the optimal incubation time (OIT) for measurement of the activated partial thromboplastin time (APTT) in dogs, rabbits, guinea pigs, rats and mice, we determined the shortest clotting time of the plasma from each animal species and compared them with that of human plasma. The OIT for APTT determination was 15 to 30 sec in guinea pigs, rats and mice and 5 to 10 minutes in dogs and rabbits. The mouse APTT (about 30 sec) with the OIT thus determined was similar to human APTT, and relatively longer than APTT in other animal species (10-20 sec). To elucidate the mechanism of the species differences in OIT, we examined the plasma of each animal species for the activity of the contact factors such as factor XII, factor XI, high molecular weight kininogen (HMWK) and prekallikrein (PK) and their effect on the coagulation of contact factor-deficient plasma. The total activity of contact factors was higher in dogs and guinea pigs and lower in rabbits and mice than that in humans. Species difference with the factor XII, Factor XI and HMWK was noted in clotting time but not in OIT. These results suggest that the species difference in OIT for APTT is probably due to difference in activity of the plasma contact factors and in the mode of coagulation for each contact factor.     

Respiratory mechanics and maximal expiratory flow in the anesthetized mouse.

Mice have been widely used in immunologic and other research to study the influence of different diseases on the lungs. However, the respiratory mechanical properties of the mouse are not clear. This study extended the methodology of measuring respiratory mechanics of anesthetized rats and guinea pigs and applied it to the mouse. First, we performed static pressure-volume and maximal expiratory flow-volume curves in 10 anesthetized paralyzed C57BL/6 mice. Second, in 10 mice, we measured dynamic respiratory compliance, forced expiratory volume in 0.1 s, and maximal expiratory flow before and after methacholine challenge. Averaged total lung capacity and functional residual capacity were 1.05 +/- 0.04 and 0.25 +/- 0.01 ml, respectively, in 20 mice weighing 22.2 +/- 0.4 g. The chest wall was very compliant. In terms of vital capacity (VC) per second, maximal expiratory flow values were 13.5, 8.0, and 2.8 VC/s at 75, 50, and 25% VC, respectively. Maximal flow-static pressure curves were relatively linear up to pressure equal to 9 cm H(2)O. In addition, methacholine challenge caused significant decreases in respiratory compliance, forced expiratory volume in 0.1 s, and maximal expiratory flow, indicating marked airway constriction. We conclude that respiratory mechanical parameters of mice (after normalization with body weight) are similar to those of guinea pigs and rats and that forced expiratory maneuver is a useful technique to detect airway constriction in this species.     

The organizational characteristics of the APUD system in mammalian lungs at different stages of ontogeny]

The organization peculiarities of APUD-system in the lungs of rabbits, rats and guinea pigs has been studied. The endocrine system in the lungs of rabbits in pre- and postnatal ontogenesis is presented by the adipocytes and neuroepithelial bodies (NEB) containing a considerable number of monoamines. The number of argyrophil adipocytes and NEBs in the lungs of 21 and more day-old adult rats seem to be less than in fetuses and newborns. Monoamines are not revealed in the endocrine rat lung structures by means of the glyoxylic acid. In the lungs of guinea pigs the single argyrophil adipocytes and NEBs are determined in the gestation period.     

Immunocytochemical studies on parafollicular cells of various mammals

Using specific antisera, calcitonin, calcitonin gene-related peptide (CGRP), somatostatin as well as neuron-specific enolase, chromogranin, secretory peptide I and calbindin (vitamin D-dependent calcium-binding protein) were looked for in parafollicular cells of rats, Syrian hamsters, Mongolian gerbils, mice, guinea pigs, rabbits and pigs. Calcitonin and CGRP were most invariably present in various species. Somatostatin was absent in mice and Mongolian gerbils and present in variable amounts in the remaining species. Neuron-specific enolase could not be detected in rabbits, while in the pigs and the Mongolian gerbils it could be demonstrated only in some parafollicular cells. Calbindin was present exclusively in parafollicular cells of guinea pigs. Chromogranin and secretory protein-I were present only in some animal species.     

Angiostrongylus cantonensis: Development following pulmonary arterial transfers into permissive and nonpermissive hosts

The survival, growth, and egg-laying capacity of young adult Angiostrongylus cantonensis, surgically transferred from intracranial sites into pulmonary arteries, were studied. A variety of experimental animals (rats, guinea pigs, mice, and mastomys) were chosen as donor animals and as recipient hosts (rats, guinea pigs, and rabbits). These species were specifically chosen to span the spectrum of host permissiveness relative to worm development in an attempt to understand the mechanisms which underlie species-dependent resistance. Recipient animals were monitored not only for the development of parasites per se but also for antibody production and histopathologic changes. The results indicated that these procedures were technically feasible, with good worm development following intra-rat transfers, as early as 15 days after initial exposure. Studies were performed to analyze the constraints of development both on initial, i.e., prelung and subsequent i.e., postlung development. When worms were obtained from permissive species such as rat or mastomys, transfer into rats resulted in good growth and development; however, worms which developed initially in exposed mice or guinea pigs developed less well in the rat. Conversely, worms which developed initially in permissive host such as the rat, when transferred into a variety of less permissive hosts such as the guinea pig and rabbit, apparently did not survive and caused significant morbidity and mortality within the nonpermissive host. Histopathologic evaluation revealed a strong eosinophilic perivascular and peribronchiolar infiltration as well as granulomatous reactions surrounding the worms in the lungs of recipient guinea pigs and rabbits, changes not observed in the lungs of permissive rat recipients. As reaginic antibody responses were also more prominent in nonpermissive than in permissive animals, it is possible that IgE responses may be more directly related to the phenomenon of morbidity and/or permissiveness than are other aspects of immune response. In support of this contention was the finding of nearly equivalent hemagglutinating antibody production between permissive rats and nonpermissive guinea pigs and rabbits.     

Species-related variations in lipoprotein metabolism: the impact of FER(HDL) on susceptibility to atherogenesis

Several animal models have been used to investigate the mechanisms of atherogenesis. Each animal species has advantages and disadvantages with regard to similarity with human lipoprotein metabolism. In humans, fractional esterification rate in apolipoprotein B-depleted plasma (FER(HDL)) has been shown to correlate with the quality of high density lipoprotein particles. Increased values of FER(HDL) indicate an atherogenic lipoprotein profile. Such an association has not been defined in animal models. Thus, we have characterized plasma lipoprotein profile and FER(HDL) values in four animal species namely, cats, pigs, guinea pigs and rabbits. These animal species have been used in experimental dyslipidemia and atherosclerosis. Our data indicate a wide rage of variations among various animal species. High-density lipoprotein (HDL) particles contain approximately 40% of total plasma cholesterol concentrations in rabbits, pigs and cats <10% in guinea pigs. A negative association between FER(HDL) values and plasma HDL-cholesterol levels was observed in pigs, rabbits and guinea pigs. On the other hand, FER(HDL) values showed a positive association with plasma triglyceride levels in all animal species tested. These findings are in agreement with data reported in humans. More research is needed to identify the better animal models which closely resemble human lipoprotein metabolism.     

Pharmacokinetics of fusidic acid in laboratory animals.

The ability of evaluate the efficacy of fusidic acid in animal models of infectious disease is limited by the absence of pharmacokinetic data for the agent in laboratory animals. In our study, aspects of fusidic acid pharmacokinetics were compared in rats (Rattus norwegicus), mice (Mus musculus), rabbits (Oryctolagus cuniculus), and guinea pigs (Cavia porcellus). Sodium fusidate was poorly absorbed after oral administration to rats, although limited absorption occurred in guinea pigs, mice, and rabbits. Subcutaneous injections of diethanolamine fusidate to laboratory rats, however, achieved a serum profile similar to that observed in humans. There was no evidence of drug accumulation in rats given repeated subcutaneous doses of diethanolamine fusidate during a 4-day period, but rabbits showed clear evidence of a cumulative effect.     

Hemoglobin and red blood cells alter the response of expired nitric oxide to mechanical forces

Expired nitric oxide (NO(e)) varies with hemodynamic or ventilatory perturbations, possibly due to shear stress- or stretch-stimulated NO production. Since hemoglobin (Hb) binds NO, NO(e) changes may reflect changes in blood volume and flow. To determine the role of blood and mechanical forces, we measured NO(e) in anesthetized rabbits, as well as rabbit lungs perfused with buffer, red blood cells (RBCs) or Hb following changes in flow, venous pressure (P(v)), and positive end-expiratory pressure (PEEP). In buffer-perfused lungs decreases in flow and P(v) reduced NO(e), but NO(e) rose when RBCs and Hb were present. These findings are consistent with changes in vascular NO production, whose detection is obscured in blood-perfused lungs by the more dominant effect of Hb NO scavenging. PEEP decreased NO(e) in all perfused lungs but increased NO(e) in live rabbits. The NO(e) fall with PEEP in isolated lungs is consistent with flow redistribution from alveolar septal capillaries to extra-alveolar vessels and decreased surface area or a direct, stretch-mediated depression of lung epithelial NO production. In live rabbits, increased NO(e) may reflect blood flow reduction and decreased Hb NO scavenging and/or autonomic responses that increase NO production. We conclude that blood and systemic responses render it difficult to use NO(e) changes as an accurate measure of lung tissue NO production.     

Role of nitric oxide during hyperventilation-induced bronchoconstriction in the guinea pig.

Airway function is largely preserved during exercise or isocapnic hyperventilation in humans and guinea pigs despite likely changes in airway milieu during hyperpnea. It is only on cessation of a hyperpneic challenge that airway function deteriorates significantly. We tested the hypothesis that nitric oxide, a known bronchodilator that is produced in the lungs and bronchi, might be responsible for the relative bronchodilation observed during hyperventilation (HV) in guinea pigs. Three groups of anesthetized guinea pigs were given saline and three groups given 50 mg/kg N(G)-monomethyl-L-arginine (L-NMMA), a potent nitric oxide synthase inhibitor. Three isocapnic ventilation groups included normal ventilation [40 breaths/min, 6 ml/kg tidal volume (VT)], increased respiratory rate only (150 breaths/min, 6 ml/kg VT), and increased respiratory rate and increased volume (100 breaths/min, 8 ml/kg VT). L-NMMA reduced expired nitric oxide in all groups. Expired nitric oxide was slightly but significantly increased by HV in the saline groups. However, inhibition of nitric oxide production had no significant effect on rate of rise of respiratory system resistance (Rrs) during HV or on the larger rise in Rrs seen 6 min after HV. We conclude that nitric oxide synthase inhibition has no effect on changes in Rrs, either during or after HV in guinea pigs.     

Experimental atherosclerosis: a historical overview

Almost one-hundred years ago the first evidence of experimental atherosclerosis was reported. Over the past century, significant advances have been made in the development of animal models of human coronary artery disease. In this minireview, induction of atherosclerotic lesions in several animal models including rodents (mice, rabbits, rats, hamsters, guinea pigs), avian (pigeons, chickens, quail), swine, carnivora (dogs, cats), and non-human primates is discussed. The limitations and advantages of the animal models of atherosclerosis have been summarized. The transgenic/knockout animal models have greatly enhanced our understanding of atherosclerosis. Compared to wild-type counterparts, the knockout/transgenic animals develop atherogenesis faster without a need for a highly atherogenic diet. Although almost all investigations support a causal role for increased plasma cholesterol levels in the development of atherosclerotic vascular disease, an increasing body of evidence indicates serious invqlvement of other factors including oxidative stress, inflammation, infection and other emerging risk factors.     

Dermatophytes isolated from laboratory animals

In order to determine the presence of dermatophytes in healthy skin, 200 animals from the animal house of Faculty of Medicine, U.N.A.M., were studied; these were 50 rats, 50 rabbits, 50 mice, and 50 guinea pigs. Out of these 200 animals, 29.5% had positive isolation of Trichophyton mentagrophytes, var. lacticolor. The frequency variation was: rats, 68%; rabbits, 36%; mice, 8%; and guinea pigs, 6%. Male rats and male rabbits, had the higher incidence of positives. The epidemiologic repercussion of these and the significance to use these animals in biomedical investigation is discussed.     

Transoral tracheal intubation of rodents using a fiberoptic laryngoscope

A fiberoptic laryngoscope which allows direct visualization of the deep pharynx and epiglottis has been developed for transoral tracheal intubation of small laboratory mammals. The device has been employed in the intubation and instillation of a variety of substances into the lungs of rats, and with minor modification, has had similar application in mice, hamsters, and guinea pigs. The simplicity and ease of handling of the laryngoscope permits one person to intubate large numbers of enflurane anesthetized animals either on an open counter top or in a glove-box, as may be required for administration of carcinogenic materials. Instillation of 7Be-labeled carbon particles into the lungs of mice, hamsters, rats, and guinea pigs resulted in reasonably consistent interlobal distribution of particles for each test animal species with minimal tracheal deposition. However, actual lung tissue doses of carbon exhibited some species dependence.     

A new conventional and high-frequency ventilator for small animals

A pressure limited, time controlled ventilator has been designed especially for studies on experimental animals with severe respiratory distress syndrome (SRDS). Inspiration: Expiration (I:E) ratio (1:99-99:1) and frequency can be changed independently. Frequency ranged from 1 to 199/min in conventional ventilation (CV), while in high-frequency jet ventilation (HFJV) from 1 to 30 Hz. The gas delivery system consists of 3 magnetic valves (inspiration, expiration and HFJV, respectively) to ensure superposition of CV with HFJ or to use them separately. A monitoring unit switches off inspiration gas sources during HFJV if intratracheal pressure exceeds the alarm threshold. The device has been used in the following animal models: premature newborn rabbits with surfactant deficient lungs, emphysematous rats and guinea pigs as well as dogs and rabbits with SRDS due to lung lavage. Ventilation was most effective with an I:E ratio of 4:1 during pressure controlled CV, whereas during HFJV optimum gas exchange could be maintained with an I:E ratio of 1:4 and a frequency of 15 Hz in beagle dogs and 10 Hz in rabbits, respectively.     

Saprophytic occurrence of Trichophyton mentagrophytes and Microsporum gypseum in the coats of healthy laboratory animals

A group of 199 healthy laboratory animals, comprising 63 guinea pigs; 58 white mice. 47 rats and 31 rabbits, was sampled for the presence of pathogenic dermatophytes. T. mentagrophytes, var. granulare, was isolated in 10% (5-guinea pigs, 6 mice, 6 rats and 2 rabbits) and M. gypseum was found in 7 animals (3 guinea pigs, 3 mice and one rat). No ringworm lesions were observed in the respective animals. This is the first report on such findings in Israel.     

Respiratory measurements of unsedated small laboratory mammals using nonrebreathing valves

The respiratory frequency, tidal volume, minute volume, oxygen uptake and carbon dioxide output of unsedated hamsters, rats, guinea pigs and rabbits were measured to obtain comparative data and to evaluate the performance of those species as unsedated subjects. The animals were trained to remain stationary and breathe through nonrebreathing valves while expired gas was collected and respiratory frequency was monitored. Measurements of dogs also were conducted to obtained comparative data by similar methods. Hamsters were readily trained and performed reliably during repeated trials. Rats and guinea pigs were more difficult to train and performed erratically. The rabbits' performance was intermediate between that of hamsters and the other species. The back pressures caused by the small animal nonrebreathing valves at estimated peak flow rates were either similar to or less than those encountered by dogs. Measured respiratory values were compared to values predicted by published equations based on body weight. Data from this study generally reflected species differences related to body weight and metabolic rate similar to those predicted by the equations, but values from the four smaller species also may have reflected differences related to behavior.     

Pathogenicity for several laboratory animals of toxoplasma oocysts originated from naturally infected cats.

The pathogenicity of four strains, O-1, O-2, O-3, and O-4, of Toxoplasma isolated in the form of oocyst from the feces of naturally infected cats was examined for such laboratory animals as mice, rats, rabbits, guinea pigs, and dogs, in comparison with that of the Beverley strain. Suspensions of seven graded doses of oocysts of each strain ranging from 1.0 X 10(-1) to 1.0 X 10(5) were inoculated orally into seven groups of five mice each. The O-1, O-2, and O-3 strains were as pathogenic for mice as the Beverley strain, but the O-4 strain was not so pathogenic as any other strain. Rats, rabbits, guinea pigs, and dogs were inoculated orally with around 1.0 X 10(5) oocysts. The four O strains were not so severly pathogenic for rats and rabbits as to cause death. The O-2 and O-3 strains showed strong pathogenicity for guinea pigs, almost all of which, when inoculated with them, died after manifesting severe clinical symptoms. The pathogenicity of the O-1 and O-2 strains showed essentially the same tendency for dogs as for any other animal. In inoculation with oocysts, as well as with proliferative forms or cysts, the same pathogenicity was not observed in different strains, even if the same species of host animals was used. On the contrary, the same pathogenicity was not always found even in one strain when a different species of animals was used.