Angiostrongylus cantonensis: Rejection of pulmonary arterial transfers of lung stage worms

Experimental transfer of the lung stage worms of Angiostrongylus cantonensis was performed between permissive hosts (rats) and between permissive (rat) and nonpermissive hosts (guinea pigs and rabbits). These worms from rats were rejected when implanted into nonpermissive hosts. Unexpectedly, similar worms did not survive well even in permissive hosts; the majority of recipient rats did not have first-stage larvae (L₁) in their stools and, even when positive for L₁, the number of the larvae shed was few. These findings contrast with the successful pulmonary arterial transfer of younger, intracranial-stage worms. It was shown that differences in rat strain between donor and recipient had no significant effect on the subsequent worm survival in recipient hosts. The alteration of maintaining conditions of the intrapulmonary worms, prior to transfer, in terms of temperature, media, and maintaining period, also showed no profound effect on the subsequent worm survival. The kinetics of precipitating and reaginic antibody levels in rats implanted with the intrapulmonary worms were analogous to those in rats with intracranial-stage worms. The findings indicate that some qualitative differences may exist between the worms obtained from two different sites.     

Eosinophil responses of permissive and nonpermissive hosts to the young adult worms of Angiostrongylus cantonensis

Blood and bone marrow eosinophilia was assessed in nonpermissive (guinea pigs) and permissive (rats) hosts following the pulmonary arterial transfers of live or dead young adult worms of Angiostrongylus cantonensis. Guinea pigs showed a marked eosinophilic response to live worms but only a slight response to dead worms. Neither IgE nor haemagglutinating antibodies correlated with the induction of this eosinophilia. In contrast, the rat responded to neither form of the young adult worm. When the guinea pig and the rat were injected with whole worm extract (WWE) of the young adult worms either by an osmotic mini-pump connected to the jugular vein or by intermittent intravenous injections, the former animal showed blood eosinophilia but the latter failed to do so. Guinea pigs also developed blood eosinophilia after continuous exposure to the excretory and secretory products of the young adult worms, administered by the mini-pump. Eosinophil responses to WWE could be induced both in athymic CD-1 (ICR) nude mice and in its heterozygous litter mates, suggesting that T cell-independent mechanism(s) could be involved in the induction of blood eosinophilia in the nonpermissive, mouse host. These data clearly indicate that the eosinophilia-inducing factor(s) and the mechanism of eosinophilia are different in permissive and nonpermissive hosts.     

Angiostrongylus cantonensis: role of eosinophils in the neurotoxic syndrome (Gordon-like phenomenon)

The role of eosinophils in the pathophysiology of Angiostrongylus cantonensis infections was investigated in nonpermissive (guinea pig) and permissive (rat) hosts. Neurological symptoms similar to the Gordon phenomenon (ataxia, tremor, paralysis) together with a loss of Purkinje cells in the cerebellum were observed after intracraneal injection of human eosinophil extracts or after infection with A. cantonensis, only in guinea pigs and not in rats. Blood eosinophilia as well as eosinophil numbers present in the cerebellum and in the cerebrospinal fluid were higher in guinea pigs than in rats, at all times after infection with A. cantonensis. Increased levels of cytotoxicity toward L3 larvae in vitro were obtained in the presence of guinea pig eosinophils and IgE antibodies, rather than with the corresponding rat effector system. The detection of one eosinophil granule component, the eosinophil peroxidase, in the cerebrospinal fluid from infected guinea pigs but not from rats suggested that in nonpermissive hosts, neurological disorders, similar to the previously described Gordon phenomenon, might be due to eosinophil neurotoxins released after interaction of eosinophils with the parasites.     

Effects of host endocrine gland removal on the permissive status of laboratory rodents to infection by Schistosoma mansoni

Knopf P. M. and Soliman M. 1980. Effects of host endocrine gland removal on the permissive status of laboratory rodents to infection by Schistosoma mansoni. International Journal for Parasitology, 10: 197–204. The capacity of Schistosoma mansoni to complete its life cycle was compared in CD-1 mice (permissive hosts) and Sprague-Dawley rats (nonpermissive hosts) from which the pituitary gland had been removed prior to infection with cercariae. Except for a modest decrease in egg burden, none of the parameters of worm life cycle assessed were affected in hypophysectomized mice. In contrast, all these parameters were affected in hypophysectomized rats, e.g. onset of adult worm elimination was delayed, worm development improved, oviposition increased and miracidia developed. Effects of removal from rats of the thyroid/parathyroid glands on the parasite life cycle were similar to hypophysectomy; adrenalectomy or gonadectomy were without affect. Differences between thyroidectomized and thymectomized rats are discussed. It is concluded that host hormones contribute to the nonpermissive status of rats to Schistosoma mansoni infections.     

The fecundity of Schistosoma mansoni in chronic nonhuman primate infections and after transplantation into naive hosts

Adult Schistosoma mansoni worms were transplanted from 8 nonhuman primates with chronic infections into 8 naive recipients, in an effort to test the hypothesis that worm fecundity reduction in chronic infections is the result of host immunity or some other host effect. Techniques for perfusing living donors without the added use of anti-schistosomal drugs and for reducing the likelihood of post-operative bacterial endotoxemia and septic shock are described. Fecundity values in terms of eggs per day per female worm were obtained for the worms in their original and in their new hosts and compared. In 3 experiments, perfusions were incomplete and the donors were saved, enabling direct comparisons of fecundity to be made in subpopulations of worms in both their original and new hosts, after equal life spans. In only 1 of the 8 transplantations was there a clear increase in fecundity after surgical introduction into a naive host. Therefore, these experiments fail to support the hypothesis that reduced fecundity of S. mansoni worms in permissive nonhuman primate hosts is a reversible result of host immunity or some other host-derived factor. Despite this negation, further evidence for reduced worm fecundity in older infections was obtained. In the absence of in vivo evidence for immune-mediated antifecundity, worm senescence is the most likely explanation for this finding, with irreversible immune damage to the worms being a less attractive alternative hypothesis.     

Host-parasite relationship of Angiostrongylus cantonensis. 1. Intracranial transplantation into various hosts.

Various stages of Angiostrongylus cantonensis recovered from the brain of experimentally infected mice were transplanted intracranially into rats. Third and fourth-stage worms recovered 2--7 days postinfection were able to develop normally after transplantation into recipient rats. The fifth-stage worm obtained 14--15 days postinfection would enter the brain tissue of rats but died shortly afterwards. However, the same stage of worms recovered from rats, after a similar transplantation, were found to develop normally in the recipient animal. Young fifth-stage worms, from the subarachnoid space of rats, which were ready for the pulmonary migration were also transplanted into rabbits but the worms failed to reach the lungs. In the control rat-to-rat transplantation, the worms successfully completed the pulmonary migration. The morphogenesis and initial growth rate of A. cantonensis were similar in both mice and rats but in the former host the worms started to grow at a markedly slower rate after the last moult and gradually degenerated.     

The role of eosinophils in Angiostrongylus cantonensis infection

Angiostrongylus cantonensis is the causative agent of human eosinophilic meningoencephalitis in the Pacific Islands and Southeast Asia. Prominent eosinophilia in the cerebrospinal fluid (CSF) of the patients has been used as one of the diagnostic criteria for the disease but the role(s) of the CSF eosinophils has remained to be elucidated. In this article, Kentaro Yoshimura, Hiroko Sugaya and Kazuto Ishido discuss the involvement of CSF eosinophils in the killing of intracranial worms and the damage of the central nervous system of the hosts, and consider why eosinophils in A. cantonensis infection play a more important role in nonpermissive hosts (including humans) than in the permissive rat host.     

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.     

Susceptibility of various animals to Pneumocystis carinii infection.

Pneumocystis carinii (Pc) is an important opportunistic pathogen of immune compromised hosts, and is known to infect various animals. The present study observed the infection status of 6 mammals and 3 strains of albino rats with Pc after suppression of their immunity. Methyl-prednisolone was injected once a week and tetracycline was supplied with water for 5 to 21 weeks. Hamsters, guinea pigs, rabbits, dogs, cats and pigs were negative by impression smear, and only the rats were found infected by Pc. All of the three strains of rats, Sprague-Dawley(SD), Wistar(W) and Fisher(F), were infected by Pc but W rats showed heavier degree of infection in earlier period than F or SD rats. The present findings suggest that W rat is the best among the animals used in the present study for production of Pc.     

Molecular Characterization of Pneumococcal Isolates from Pets and Laboratory Animals

Background

Between 1986 and 2008 Streptococcus pneumoniae was isolated from 41 pets/zoo animals (guinea pigs (n = 17), cats (n = 12), horses (n = 4), dogs (n = 3), dolphins (n = 2), rat (n = 2), gorilla (n = 1)) treated in medical veterinary laboratories and zoos, and 44 laboratory animals (mastomys (multimammate mice; n = 32), mice (n = 6), rats (n = 4), guinea pigs (n = 2)) during routine health monitoring in an animal facility. S. pneumoniae was isolated from nose, lung and respiratory tract, eye, ear and other sites.

Methodology/Principal Findings

Carriage of the same isolate of S. pneumoniae over a period of up to 22 weeks was shown for four mastomys. Forty-one animals showed disease symptoms. Pneumococcal isolates were characterized by optochin sensitivity, bile solubility, DNA hybridization, pneumolysin PCR, serotyping and multilocus sequence typing. Eighteen of the 32 mastomys isolates (56%) were optochin resistant, all other isolates were optochin susceptible. All mastomys isolates were serotype 14, all guinea pig isolates serotype 19F, all horse isolates serotype 3. Rats had serotypes 14 or 19A, mice 33A or 33F. Dolphins had serotype 23F, the gorilla serotype 14. Cats and dogs had many different serotypes. Four isolates were resistant to macrolides, three isolates also to clindamycin and tetracyclin. Mastomys isolates were sequence type (ST) 15 (serotype 14), an ST/serotype combination commonly found in human isolates. Cats, dogs, pet rats, gorilla and dolphins showed various human ST/serotype combinations. Lab rats and lab mice showed single locus variants (SLV) of human STs, in human ST/serotype combinations. All guinea pig isolates showed the same completely new combination of known alleles. The horse isolates showed an unknown allele combination and three new alleles.

Conclusions/Significance

The isolates found in mastomys, mice, rats, cats, dogs, gorilla and dolphins are most likely identical to human pneumococcal isolates. Isolates from guinea pigs and horses appear to be specialized clones for these animals. Our data redraw attention to the fact that pneumococci are not strictly human pathogens. Pet animals that live in close contact to humans, especially children, can be infected by human isolates and also carriage of even resistant isolates is a realistic possibility.     

CD81 expression is important for the permissiveness of Huh7 cell clones for heterogeneous hepatitis C virus infection

Huh7 cells constitute a permissive cell line for cell culture of hepatitis C virus (HCV) particles. However, our Huh7 line shows limited permissiveness for HCV. Thus, in this study we set out to determine which host factors are important for conferring permissiveness. To analyze the limited permissiveness of our Huh7 cells, 70 clones were obtained after single-cell cloning of parental Huh7 cells. The cloned Huh7 cells exhibited various levels of HCV pseudoparticles and JFH-1 virus infection efficiency, and some clones were not permissive. A subgenomic replicon was then transfected into the cloned Huh7 cells. While the replication efficiencies differed among the cloned Huh7 cells, these efficiencies did not correlate with infectious permissibility. Flow cytometry showed that CD81, scavenger receptor class B type I, and low-density-lipoprotein receptor expression on the cell surfaces of the Huh7 clones differed among the clones. Interestingly, we found that all of the permissive cell clones expressed CD81 while the nonpermissive cell clones did not. To confirm the importance of CD81 expression for HCV permissiveness, CD81 was then transiently and stably expressed on a nonpermissive Huh7 cell clone, which was consequently restored to HCV infection permissiveness. Furthermore, permissiveness was down-regulated upon transfection of CD81 silencing RNA into a CD81-positive cell clone. In conclusion, CD81 expression is an important determinant of HCV permissiveness of Huh7 cell clones harboring different characteristics.     

Life cycle and host specificity of Amblyomma triste (Acari: Ixodidae) under laboratory conditions

We report biological data of two generations of Amblyomma triste in laboratory and compared the suitability of different host species. Infestations by larval and nymphal stages were performed on guinea pigs (Cavia porcellus), chickens (Gallus gallus), rats (Rattus norvegicus), rabbits (Oryctolagus cuniculus), wild mice (Calomys callosus), dogs (Canis familiaris) and capybaras (Hydrochaeris hydrochaeris). Infestations by adult ticks were performed on dogs, capybaras and rabbits. Tick developmental periods were observed in an incubator at 27 degrees C and RH 90%. Guinea pigs were the most suitable hosts for larvae and nymphs, followed by chickens. The remaining host species were less suitable for immature ticks as fewer engorged ticks were recovered from them. Mean larval feeding periods varied from 3.8 to 4.7 d between different host species. Mean larval premolt periods ranged from 8.9 to 10.4 d. Nymphal mean feeding periods varied from 4.2 to 6.2 d for ticks fed on different host species. Premolt period of male nymphs (mean: 15.4 d) was significantly longer than that of female nymphs (14.7 d). Female nymphs were significantly heavier than male nymphs. The overall sex ratio of the adult ticks emerged from nymphs was 0.9:1 (M:F). Capybaras were the most suitable host for the tick adult stage as significantly more engorged females were recovered from them and these females were significantly heavier than those recovered from dogs or rabbits. The life cycle of A. triste in laboratory could be completed in an average period of 155 d. The potential role of guinea pigs, birds and capybaras, as hosts for A. triste in nature, is discussed.     

Surface antigens of Angiostrongylus cantonensis developing in permissive and non-permissive hosts

The immunofluorescent antibody test and immunocytochemical method were employed to study the surface antigens of Angiostrongylus cantonensis obtained from infected rats, mice and guinea pigs. Positive results with intense fluorescence and brownish peroxidase staining were observed on the cuticular surface of A. cantonensis recovered from rats 22 days (late cerebral phase) and 34 days (lung phase) post-infection when tested with antisera against host (normal rat serum) antigens as well as crude extracts of A. cantonensis. However, host antigens were not observed on the surface of the nematode recovered from the brain of mice and guinea pigs 15 days post-infection.     

Endemic echinostome infections of candidate hosts

Wild Lymnaea tomentosa snails, recovered from Lake Wanaka, New Zealand, were established in the laboratory. Wild snails, naturally infected with echinostomes, provided metacercariae for infection of laboratory maintained snails. Metacercarial cysts from wild and laboratory snails were then used to attempt infection of definitive host candidates. Laboratory snails provided convenient packaging of known numbers of cysts. Metacercariae excysted in the small intestines of ducklings to mature in 6 days. Worms were expelled as they became gravid. Attempts to establish infections in experimental hosts other than ducklings were not successful. No worms were recovered from mice, white rats, guinea pigs, hamsters or immunosuppressed white rats.     

Complement requirement of neutralizing antibodies in different classes of immunoglobulin appearing in rabbits and guinea pigs after primary and booster immunizations with herpes simplex virus.

Rabbits and guinea pigs were immunized with herpes simplex virus and bled periodically. The sera were fractionated into slow IgG, fast IgG and IgM by DEAE-cellulose column chromatography, and complement-requiring (CRN) and nonrequiring neutralizing (N) antibody activities were estimated. In early sera of rabbits, the two IgG and IgM fractions possessed about equal CRN activities, although some animals showed a slightly lower activity in fast IgG. In guinea pigs, the early CRN activity resided mainly in slow IgS (7 S gamma2). The early IgG antibody of guinea pigs differed from that of rabbits in that it resembled IgM in resistances to heating at 70 C and to 2-merceptoethanol. The level of CRN IgM antibody in rabbits declined following a peak reached in 2 to 3 weeks, whereas such a decline was never observed in guinea pigs. N IgG antibody was developed a few weeks after the first immunization in rabbits and much retarded in guinea pigs. In both species, booster immunization quickly evoked N antibody in the two IgG fractions and also CRN IgM antibody, but in the case of rabbits the IgM antibody disappeared soon. It is concluded that IgG plays an important role in humoral immunity from the initial stage of the immunization course.     

Complement Requirement of Neutralizing Antibodies in Different Classes of Immunoglobulin Appearing in Rabbits and Guinea Pigs after Primary and Booster Immunizations with Herpes Simplex Virus

Rabbits and guinea pigs were immunized with herpes simplex virus and bled periodically. The sera were fractionated into slow IgG, fast IgG and IgM by DEAE-cellulose column chromatography, and complement-requiring (CRN) and nonrequiring neutralizing (N) antibody activities were estimated. In early sera of rabbits, the two IgG and IgM fractions possessed about equal CRN activities, although some animals showed a slightly lower activity in fast IgG. In guinea pigs, the early CRN activity resided mainly in slow IgG (7 S γ₂). The early IgG antibody of guinea pigs differed from that of rabbits in that it resembled IgM in resistances to heating at 70 C and to 2-mercaptoethanol. The level of CRN IgM antibody in rabbits declined following a peak reached in 2 to 3 weeks, whereas such a decline was never observed in guinea pigs. N IgG antibody was developed a few weeks after the first immunization in rabbits and much retarded in guinea pigs. In both species, booster immunization quickly evoked N antibody in the two IgG fractions and also CRN IgM antibody, but in the case of rabbits the IgM antibody disappeared soon. It is concluded that IgG plays an important role in humoral immunity from the initial stage of the immunization course.     

Strongyloides ratti: reversibility of immune damage to adult worms

Transplantation experiments were conducted to assess the reversibility or irreversibility of the damage sustained by Strongyloides ratti during infections in the rat host. Worms of different ages from primary and secondary infections were recovered from their original hosts and transplanted surgically into naive rats. The size and fecundity of normal (Days 6–11 postinfection) worms were maintained after transfer. Damaged worms from primary infection (Days 22–26) showed complete recovery of size and fecundity within 10 days of transfer; damaged worms from a secondary infection (Days 6–7) also showed functional recovery but to a lesser extent. The ultrastructural changes observed mainly in the intestine of damaged worms from primary infections, prior to their transfer, were, however, only partially ameliorated following transplantation into new naive hosts; there was no complete return to structural normality. On the other hand, second infection worms did show almost complete ultrastructural recovery. The course of a transplanted infection established with either damaged or normal worms was similar to infections established percutaneously. Increase in the size of transplanted infections from 100 to 250 worms per recipient did not alter the dynamics of the host/parasite relationship. There was no evidence of adaptation in S. ratti and damaged worms, when transplanted into naive rats, were as successful as normal worms in protecting the host against a subcutaneous larval infection. The implications of this work on the present understanding of the phenomenon of autoinfection in experimental rodent strongyloidiasis are discussed.     

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)     

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.     

A heterophile antigen associated with experimental toxoplasmosis

The biological characteristics of a heterophile protein (HP) in peritoneal exudate from mice, hamsters, rats, and guinea pigs infected with Toxoplasma gondii were studied by immunofluorescence, immunoelectrophoresis and immunodiffusion techniques using specific antisera raised in rabbits. HP of mice had the highest antigenicity, HP of hamsters and rats had intermediate antigenicity and HP of guinea pigs had the lowest antigenicity. HP was found in normal peritoneal exudates from mice, hamsters and rats inoculated with paraffin oil instead of T. gondii and in normal guinea pig serum. HP was detected by the fluorescent antibody technique on the surface of T. gondii in peritoneal exudates of mice, but not on mouse peritoneal cells, and by the indirect fluorescent antibody technique on L cells infected with T. gondii and on free Toxoplasma derived from them, but not on uninfected L cells. T. gondii could make host cells produce HP to cover its surface for protection. The relation between HP from host cells and T. gondii is discussed.