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Trypanosoma cruzi in the anal glands of urban opossums. I--Isolation and experimental infections.

Opossums (Didelphis marsupialis) captured in intensely urbanized areas of the city of Caracas, Venezuela, were found infected with Trypanosoma cruzi. The developmental cycle of trypomastigote-epimastigote-metacyclic infective trypomastigote, usually occurring in the intestine of the triatomine vector, was taking place in the anal odoriferous glands of the opossums. Material from the glands, inoculated in young, healthy opossums and white mice by different routes, subcutaneously, intraperitoneally, orally, and into the eye, induced T. cruzi infections in all animals. Parasitemia, invasion of cardiac and skeletal muscle, and intracellular multiplication of amastigotes were observed. Inoculation of metacyclics from anal glands, cultured in LIT medium, gave equivalent results. All opossums survived; all mice died. Excreta of opossums may thus transmit Chagas' disease by contamination, even in urban areas where insect vectors are not present.

Anal Canal↗
PubMed2002

The opossum Didelphis virginiana as a synanthropic reservoir of Trypanosoma cruzi in Dzidzilché, Yucatán, México.

In México, the role of mammals in the transmission cycle of Trypanosoma cruzi is poorly known. In the State of Yucatán, an endemic area of Chagas disease, both Didelphis virginiana and D. marsupialis occur sympatrically. However, until now, only the former species had been found infected with T. cruzi. To evaluate the role of D. virginiana in a peridomestic transmission, nine periods of capture-recapture were performed around the village of Dzidzilché, Yucatán. The sex, age, reproductive status, location, and presence of infection with T. cruzi were recorded for each opossum. The chromosome morphology was used to identify the opossum species. T. cruzi was identified by the presence of pseudocysts of amastigotes in cardiac muscle fibers of Balb/c mice inoculated with strains isolated from opossums. However, xenodiagnosis was the best diagnostic method. Triatoma dimidiata, the vector, were collected in and around the opossums' nests, and human dwellings; and were checked for T. cruzi. From 102 blood samples of D. virginiana examined 55 (53.9%) were positive to T. cruzi, the only two D. marsupialis captured were negative. Significant differences were found between infection, and both sex and reproductive condition. Eight out of 14 triatomines collected in peridomestic nests (57.1%), and 32 of 197 captured inside houses (16.3%) were found infected, suggesting a peridomestic transmission. The statistically high abundance of infected opossums and triatomines during the dry season (March to May) suggested the existence of a seasonality in the peridomestic transmission of T. cruzi in Dzidzilché.

Animals↗
PubMed1999

[Prevalence of Trypanosoma cruzi infection in opossum (Didelphis albiventris) in Santiago del Estero, Argentina].

The opossum of the genus Didelphis is one of the principal wild reservoirs of Trypanosoma cruzi and is widely distributed in the Western Hemisphere. Didelphis albiventris is the most common marsupial in Amamá and Trinidad, two communities in the province of Santiago del Estero, Argentina. The D. albiventris population is replaced every year, and the opossum normally has two reproductive periods, one at the beginning of the spring and another at the beginning of the summer. The two litters are weaned, and they leave the mother's marsupial pouch to join the population, the first (G1) at the beginning of the summer and the second (G2) at the beginning of the fall. Between 1988 and 1991 409 D. albiventris opossums were studied, and xenodiagnoses showed that 35% of them were infected with T. cruzi. Annual cycles of renewed infection were observed, with prevalences that ranged between 22% and 43%. The acquisition of the parasite occurred over the entire year, from the summer through the spring. The prevalence of infection increased with age. The G1 individuals tended to present higher prevalences than the G2 individuals, probably from being exposed to transmission for a longer period of time. In the first two (younger) age categories for the opossums, G2 individuals showed higher prevalences than did the G1 individuals. This indicates a significant increase in transmission intensity during the fall. Opossums should be regarded as a potential source of T. cruzi entry to the domestic transmission cycle.

Animals↗
PubMed1993

A suburban focus of endemic typhus in Los Angeles County: association with seropositive domestic cats and opossums.

Thirty-three cases of locally acquired murine typhus were reported in Los Angeles County residents from May 1984 through February 1988. Only eight cases were reported over the previous 20-year period. Thirty (91%) cases resided within a suburban area encompassing approximately 50 km2 in northcentral Los Angeles or had contact with an animal from this area. Serologic testing (complement fixation and indirect fluorescent antibody) of selected animals in close association with human cases revealed a high prevalence of seropositivity among domestic cats and opossums. Nine (90%) of 10 resident cats tested had demonstrable antibody titers compared with none (0%) of 20 cats from a control area (P < 0.001). Suburban typhus cases were more likely than neighborhood controls to own a cat or dog (odds ratio = 6.9, 95% confidence interval = 1.8, 25.9, P = 0.002). Sixteen (42%) of 38 opossums trapped in close proximity to the residences of cases were seropositive versus none (0%) of 36 opossums from control areas (P < 0.001) A low frequency (2.8%) of seropositivity was found in commensal rodents, and the classic vector of murine typhus, Xenopsylla cheopis, was not found. Ectoparasite indices form seropositive opossums revealed heavy infestations with the cat flea, Ctenocephalides felis (mean flea count = 104.7), a species that readily bites humans. These data provide evidence that a suburban focus of murine typhus exists in Los Angeles that differs substantially from the classic transmission cycle, and that cats, opossums and C. felis may play an important role in the occurrence of human cases.

Adolescent↗
PubMed1988

Experimental Cryptosporidium parvum infections in opossums (Didelphis virginiana).

Five nursing opossums (Didelphis virginiana) were each inoculated with 5 x 10(6) Cryptosporidium parvum oocysts of calf origin. Following inoculation, endogenous stages of C. parvum were observed in the ileum, cecum, and colon of these opossums. Two of three noninoculated pouch mates acquired infections during the study based on examinations of feces and tissue sections of all eight opossums. Mild diarrhea was observed in four of seven opossums harboring C. parvum, although none died as a result of the infection. Under the conditions of this study, C. parvum appeared to be only mildly pathogenic for opossums.

Animals↗
PubMed2000

Granulomatous pneumonia in the opossum (Didelphis virginiana) associated with an intracellular fungal agent.

BACKGROUND AND PURPOSE: Two of nine female opossums (Didelphis virginiana) in a closed breeding colony were submitted for necropsy due to a history of poor reproductive performance in the absence of overt clinical disease. On histologic examination, marked granulomatous to pyogranulomatous pneumonia was identified in these animals. METHODS: Lung sections were stained with periodic acid-Schiff and Gomori's methenamine silver nitrate. RESULTS: Pulmonary lesions were characterized by large numbers of foamy macrophages within the alveoli and interstitium, prominent subpleural and peribronchiolar aggregates of histiocytes, and a few scattered lymphoid nodules. Numerous fungal organisms were evident within the cytoplasm of macrophages on impregnation of histologic sections with the aforementioned stains. Other inciting agents were not identified. A third opossum lacked pulmonary lesions, but had similar organisms within one auricular sebaceous gland/hair follicle without apparent reaction to the organisms. CONCLUSION: A fungal agent was associated with granulomatous pneumonia in the opossum, and comparison was made with endogenous lipid pneumonia previously described in opossums. These findings stress the importance of use of special stains and additional diagnostic techniques when prominent alveolar macrophage accumulation is present on histologic examination of the opossum lung.

Animals↗
PubMed1979

Nitric oxide induced conformational changes in opossum hemoglobin.

Opossum hemoglobin assumes a T quaternary structure upon NO ligation in the absence of organic phophates at pH 6.7. In addition, stripped opossum hemoglobin exhibits a low oxygen affinity when compared to human hemoglobin and a pH-dependent heme-heme interaction with an n value of 2.14 at pH 7.0 and 2.46 at pH 7.35. These observations indicate that opossum hemoglobin may have a destabilized oxy structure when compared to hemoglobin A due to differences in primary structure. Thus, the strong trans ligand effect of nitric oxide is able to disrupt the proximal histidine-iron bond in the alpha-hemes triggering a conformational transition to the T state. Absence of a distal histidine in the alpha-subunits and, therefore an impaired donor acceptor interaction with the sixth ligand, could contribute to the lack of stability of the R quaternary structure in opossum nitrosylhemoglobin. The reduced oxygen affinity of opossum hemoglobin may be compensated for by other physiological factors such as a reduced phosphate effect.

Amino Acids↗
PubMed2002

Origins of primate locomotion: gait mechanics of the woolly opossum.

The locomotion of primates differs from that of other mammals in three fundamental ways. During quadrupedal walking, primates use diagonal sequence gaits, protract their arms more at forelimb touchdown, and experience lower vertical substrate reaction forces on their forelimbs relative to their hindlimbs. It is widely held that the unusual walking gaits of primates represent a basal adaptation for movement on thin, flexible branches and reflect a major change in the functional role of the forelimb. However, little data on nonprimate arboreal mammals exist to test this notion. To that end, we examined the gait mechanics of the woolly opossum (Caluromys philander), a marsupial convergent with small-bodied prosimians in ecology, behavior, and morphology. Data on the footfall sequence, relative arm protraction, and peak vertical substrate reaction forces were obtained from videotapes and force records for three adult woolly opossums walking quadrupedally on a wooden runway and a thin pole. For all steps recorded on both substrates, woolly opossums always used diagonal sequence walking gaits, protracted their arms beyond 90 degrees relative to horizontal body axis, and experienced peak vertical substrate reaction forces on forelimbs that were significantly lower than on hindlimbs. The woolly opossum is the first nonprimate mammal to show locomotor mechanics that are identical to those of primates. This case of convergence between primates and a committed fine-branch, arboreal marsupial strongly implies that the earliest primates evolved gait specializations for fine-branch locomotion, which reflect important changes in forelimb function.

Animals↗
PubMed1984

Cerebellar cortical efferent fibers in the North American opossum, Didelphis virginiana. I. The anterior lobe.

The distribution of cerebellar corticonuclear and corticovestibular fibers from the anterior lobe of the North American opossum, Didelphis virginiana, was studied using the Fink and Heimer ('67) technique. Corticonuclear fibers from medial areas of anterior lobe project into the medial cerebellar nucleus (NM) in a topographically organized manner. Fibers from lobules II and III enter rostral and rostrodorsal NM, while those from lobules IV and V terminate in progressively more caudal parts of the nucleus. Collectively the terminal fields in NM for axons from lobules II-V occupy about the rostral two-thirds of the nucleus. Those areas of lobules II-V of opossum that project into NM presumably correspond to zone A of cat and primate. Cerebellar corticovestibular fibers originate from cortex located immediately lateral to areas projecting to NM. The predominance of corticovestibular projections into the lateral vestibular nucleus suggests the presence of a B zone and identifies its points of interface with the A zone. The results further suggest that zones A and B overlap at their respective margins. In contrast to other mammals studied to date, zones A and B of opossum anterior lobe are comparatively wide. Corticonuclear fibers to anterior and posterior interposed nuclei and to the lateral cerebellar nucleus (NL) originate from relatively narrow lateral portions of anterior lobe. These results also suggest that the intermediate cortex of opossum anterior lobe is not clearly divisible into individual zones C1,C2 or C3. The cortical area that innervates NL is very narrow and presumably corresponds to zone D of other forms.

Animals↗
PubMed1986

Origin of mammalian thalamocortical projections. I. Telencephalic projections of the medial geniculate body in the opossum (Didelphis virginiana).

Telencephalic projections from the medial geniculate nucleus (MG) in opossum were traced with tritiated leucine autoradiography and by horseradish peroxidase and fluorescent dye retrograde labeling techniques. The results show that the opossum's MG contains two separate populations of neurons-one in the anterior two-thirds of MG projecting to auditory neocortex, the other occupying the entire caudal one-third of MG and projecting mostly to lateral amygdala and putamen. Because the subcortical projection of the MG in opossum is larger than that seen in any other mammal to date, it is reminiscent of the subcortical projections of the MG in reptiles and birds. Furthermore, when the subcortical projections of the MG in reptiles and opossums are compared with similar subcortical projections of the MG in rats, cats, and monkeys, the proportion of the MG neurons projecting to subcortical structures is seen to be inversely related to the recency of each animal's common ancestry with primates. The possibility that the subcortical projection of the MG in mammals is homologous with that seen in reptiles or birds implies that it might be a dwindling vestige of the projection present in the common ancestry of reptiles and mammals.

Animals↗
PubMed1994

Developmental distribution of GFAP and vimentin in the Brazilian opossum brain.

Cells of glial origin are involved in the morphogenesis of the mammalian central nervous system (CNS). Characterization of glial-associated proteins during neurogenesis and differentiation may aid in understanding the complexity of CNS development. We have utilized immunoblotting and immunohistochemistry to characterize the developmental profiles of glial fibrillary acidic protein (GFAP) and vimentin (VIM) in the brain of the Brazilian opossum, Monodelphis domestica. Typical of marsupials, CNS morphogenesis and neurogenesis in the opossum extend well into the postnatal period. Opossum GFAP and VIM were found as single bands at molecular weights consistent with those reported for other species, thus indicating conservation of the VIM and GFAP proteins through mammalian evolution. Differential developmental trends were observed for both proteins with relative VIM levels decreasing and GFAP levels increasing with age. Vimentin-like immunoreactivity (VIM-IR) was present at day 1 of postnatal life throughout the brain. The density of VIM-IR was maximal at 10 and 15 days postnatal (especially in radial glial elements) and decreased slightly by 25 days postnatal. In the adult brain, VIM-IR was markedly reduced compared to that of younger ages. In contrast, GFAP-like immunoreactivity (GFAP-IR) in the brain of Monodelphis increased dramatically with age. No GFAP-IR was observed in the 1 and 5 day postnatal brains. By 25 days postnatal, the pattern of GFAP-IR in the brainstem resembled that of the adult. In the forebrain, more GFAP-IR was present than at younger ages. The adult distribution of GFAP-IR was very similar to that reported for other mammalian species. These results indicate that GFAP and VIM are reciprocally related during periods of morphogenesis and differentiation of the opossum brain.

Aging↗
PubMed1990

Immunohistochemical localization of cholecystokinin in the medial preoptic area and anterior hypothalamus of the Brazilian gray short-tailed opossum: a sex difference.

We have studied the anatomical localization of cholecystokinin-like immunoreactivity (CCK IR) in somata and fibers in the medial preoptic area (MPA) and anterior hypothalamus (AH) of the Brazilian gray short-tailed opossum, (Monodelphis domestica). With the aid of an avidin-biotin, nickel-enhanced, immunohistochemical technique, CCK IR neuronal elements were found within the MPA and AH. A large number of CCK IR cell bodies were located in the MPA of colchicine-treated opossums. The MPA also contained a CCK IR fiber plexus. Quantitative image analysis revealed that the periventricular preoptic area of noncolchicine-treated male opossums had a significantly higher percent of blocked light measurements than that of the noncolchicine-treated females, indicating a higher density of CCK IR neuronal elements in the males. Neuronal fibers and somata containing CCK IR were also found within the periventricular hypothalamic nucleus (Pe), and the suprachiasmatic nucleus (SCh). These results show that CCK IR neuronal elements are found within the MPA and AH of the Brazilian short-tailed opossum. Furthermore, there is a sexually dimorphic distribution of CCK IR elements within the MPA of this small marsupial.

Animals↗
PubMed1999

Marsupial light chains: IGK with four V families in the opossum Monodelphis domestica.

A full-length and several partial cDNAs encoding IGK light chains from the marsupial South American opossum, Monodelphis domestica, were isolated and characterized. Using these clones as a starting point, the expressed IGKV repertoire was sampled by anchored polymerase chain reaction using an IGKC-specific primer. Based on nucleotide sequences of twenty unique, expressed IGKV-J combinations, there are at least four IGKV families and two J segments. Southern blot analysis revealed each IGK-V family contains multiple gene segments totaling at least thirty-five IGKV in the opossum genome. No evidence for particular, recurrent IGKV-J combinations in the opossum IGK repertoire was seen, rather the V-J combinations appeared random and diverse. Each of the four IGKV families appear more closely related to V segments from placental mammals than to each other, suggesting the duplication of the IGKV families prior to the separation of marsupials and placental mammals more than one-hundred-million years ago. Overall, the complexity of opossum light chain V segments appears greater than that found in the heavy chain, and light chains are likely to contribute significantly to Ig diversity in this species. With this report, the homologues encoding all three classes of eutherian Ig chains, IGH, IGL, and IGK, have been described in a non-placental mammal.

Amino Acid Sequence↗
PubMed1996

Bronchial circulation in the marsupial opossum, Didelphis marsupialis.

This study characterizes the existence of a bronchial circulation in a marsupial, an animal which does not undergo placental development and does not have a ductus arteriosus. Direct perfusion of the lung by the pulmonary vasculature during the fetal development of opossums may occur, potentially eliminating the need for a bronchial circulation. We used radio- and fluorescent-labeled microspheres in conjunction with postmortem intravascular casting to determine if opossums have a systemic (bronchial) blood supply to the lung (n = 9). Gross postmortem examination of the intravascular casts showed a well-developed common bronchial artery. The histological distribution pattern of fluorescent microspheres was primarily to the airways. A few fluorescent microspheres were observed in the alveolar capillaries, indicating that a precapillary bronchial-to-pulmonary anastomosis exists in the opossum. Using the reference flow technique, total bronchial blood flow to the left lung averaged 0.95 +/- 0.58 SE ml/min. The presence of a bronchial circulation in the opossum suggests that it is more than a vestigial structure from embryonic development, potentially supporting its functional importance for carrying nutrients to the airway.

Animals↗
PubMed1981

The opossum, Didelphis marsupialis (Marsupialia: Didelphidae), as a reservoir host of Leishmania braziliensis guyanensis in the Amazon Basin of Brazil.

A total of 52 opossums (six species) were examined for evidence of infection with Leishmania in three different areas of forest near Manaus, Amazonas State, Brazil. No infections were detected in 27 opossums from a region of relatively undisturbed forest, including specimens of Didelphis marsupialis (18); Metachirus nudicaudatus (four); Monodelphis brevicaudata (one); Marmosa cinerea (two); M. murina (one) and M. parvidens (one). Of 15 D. marsupialis captured from a biological reserve, much disturbed by man, three were infected with L. braziliensis guyanensis: isolations were made from the skin of two of the animals, and from the viscera of the third. The isolates were biologically and biochemically indistinguishable from one isolate of L. b. guyanensis made from man and two from the sandfly vector Lutzomyia umbratilis from the same area. Two of eight D. marsupialis and both of two M. cinerea from another area of virgin forest used for army manoeuvres were infected with Leishmania mexicana amazonensis: the parasite was in all four cases isolated from normal skin. Five of nine specimens of Proechimys guyannensis, from the vicinity of Manaus, were also infected with L. m. amazonensis. A further 13 mammals (eight species) were negative for Leishmania. The importance of opossums as a reservoir of L. b. guyanensis is discussed. Although they may play only a minor role in virgin forest which is undisturbed by man, opossums (D. marsupialis) may become a significant reservoir of infection where man's activities have eliminated the major reservoir--which has yet to be incriminated.

Animals↗
PubMed1989

The early development of major projections to the dorsal striatum in the North American opossum.

We have employed immunocytochemical and axonal transport techniques to study the development of major projections to the dorsal striatum of the North American opossum. The opossum is born in a very immature state, 12-13 days after conception, and climbs into an external pouch where it remains attached to a nipple for several months. Its immaturity at birth and its protracted postnatal development make the opossum a good model for developmental studies. Although tyrosine hydroxylase-like immunoreactive (TH-LI), presumably dopaminergic, neurons were present in the ventral mesencephalon at birth (the presumptive substantia nigra and ventral tegmental area), there was no evidence for TH-LI axons in the striatal anlage. By postnatal day (PD)6, a few immunostained axons were found within the putamen. The subsequent growth of TH-LI axons into the striatum followed general caudal to rostral and ventrolateral to dorsomedial gradients and, at any age, they were most numerous in the areas exhibiting the greatest cytodifferentiation. By estimated (E)PD45, TH-LI axons were present in most, if not all, areas of the striatum. Serotoninergic (5-HT)-LI axons were found lateral to the presumptive striatum at birth but not within it. By PD7, however, a few 5-HT-LI axons could be identified in the putamen. The growth of 5-HT-LI axons into the striatum generally followed the same gradients described for TH-LI axons although at all ages their density was much less. Using the orthograde transport of wheat germ agglutinin conjugated to horseradish peroxidase (WGA-HRP), evidence was obtained for the existence of thalamostriatal projections by PD5 and for corticostriatal projections by PD10. Crossed corticostriatal projections were present by EPD23. Our results suggest that the development of major projections to the striatum occurs postnatally in the opossum, rather than prenatally as in placental animals. The timetable for striatal innervation is discussed in light of the developmental sequences established for other motor circuits.

Animals↗
PubMed1991

Ontogeny of cholecystokinin-like immunoreactivity in the Brazilian opossum brain.

We have studied the anatomical distribution of cholecystokinin-like immunoreactive (CCK-IR) somata and fibers in the brain of the adult and developing Brazilian short-tailed opossum, Monodelphis domestica. Animals ranged in age from the day of birth (1PN) to young adulthood (180PN). A nickel enhanced, avidin-biotin, indirect immunohistochemical technique was used to identify CCK-IR structures. Somata containing CCK immunoreactivity were observed in the cerebral cortex, hippocampus, hypothalamus, thalamus, midbrain, and brainstem in the adult. Cholecystokinin immunoreactive fibers had a wide distribution in the adult Monodelphis brain. The only major region of the brain that did not contain CCK-IR fibers was the cerebellum. The earliest expression of CCK immunoreactivity was found in fibers in the dorsal brainstem of 5-day-old opossum pups. It is possible that the CCK-IR fibers in the brainstem at 5PN are of vagal origin. Cholecystokinin immunoreactive somata were observed in the brainstem on 10PN. The CCK-IR cell bodies observed in the brainstem at 10PN may mark the first expression of CCK-IR elements intrinsic to the brain. A broad spectrum of patterns of onset of CCK expression was observed in the opossum brain. The early occurrence and varied ontogenesis of CCK-IR structures indicates CCK may be involved in the function of a variety of circuits from the brainstem to the cerebral cortex. The early expression of CCK-IR structures in the dorsal brainstem suggests that CCK may modulate feeding behavior in the Monodelphis neonate. Cholecystokinin immunoreactivity in forebrain structures such as the suprachiasmatic nucleus, medial preoptic area, thalamus and cortical structures indicates that CCK may also be involved in circadian rhythmicity, reproductive functions, as well as the state of arousal of the Brazilian opossum. The ontogenic timing of CCK immunoreactivity in specific circuitry also indicates that CCK expression does not occur simultaneously throughout the brain. This pattern of CCK onset may relate to the temporal need for CCK in specific circuits of the central nervous system (CNS) during development.

Aging↗
PubMed1992

Galanin-like immunoreactivity in the adult and developing Brazilian opossum brain.

The distribution of galanin-like immunoreactivity has been characterized in the brain of the adult and developing Brazilian opossum (Monodelphis domestica). Two commercially available antisera were used to examine the distribution of galanin-like immunoreactive (GAL-IR) cells and fibers. Nuclear groups containing GAL-IR cell bodies and fibers were seen throughout the adult opossum brain. The distribution of GAL-IR elements seen is similar to that reported for other mammals. Based on these findings, we believe that galanin may have similar physiological functions in the adult Brazilian opossum as has been reported for other mammals. In the developing brain, GAL-IR structures were seen as early as 1 day postnatal (PN) in the developing hypothalamus and brainstem. By days 5 and 10 PN, there was a robust expression of galanin-like immunoreactivity in specific regions of the brain. Since neurogenesis and brain morphogenesis are actively occuring postnatally in the opossum, galanin may be playing a role in the differentation of specific regions of the brain.

Aging↗