Characterization of a virulent Leptospira interrogans strain isolated from an abandoned swimming pool

Pathogenic Leptospira spp. are the etiological agents of leptospirosis, an important disease of both humans and animals. In urban settings, L. interrogans serovars are the predominant cause of disease in humans. The purpose of this study was to characterize a novel Leptospira isolate recovered from an abandoned swimming pool. Molecular characterization through sequencing of the rpoB gene revealed 100% identity with L. interrogans and variable-number tandem-repeat (VNTR) analysis resulted in a banding pattern identical to L. interrogans serogroup Icterohaemorrhagiae, serovar Copenhageni or Icterohaemorrhagiae. The virulence of the strain was determined in a hamster model of lethal leptospirosis. The lethal dose 50% (LD50) was calculated to be two leptospires in female hamsters and a histopathological examination of infected animals found typical lesions associated with severe leptospirosis, including renal epithelium degeneration, hepatic karyomegaly, liver-plate disarray and lymphocyte infiltration. This highly virulent strain is now available for use in further studies, especially evaluation of vaccine candidates.(AU)

Effects of diazepam on Mycobacterium bovis-induced infection in hamsters

The in utero exposure of hamsters to low doses of diazepam results in impaired host defense against Mycobacterium bovis during adulthood. Delayed developmental immunotoxicity, however, represents a specific situation that might not be general. The present experiment was undertaken to investigate the effects of diazepam on hamster resistance to M. bovis using adult animals. The effects of diazepam treatment on serum cortisol levels were also studied. Adult hamsters (N = 10 for each group) were treated with diazepam (E1 = 1.0, E2 = 2.0 or E3 = 3.0 mg kg-1 day-1 subcutaneously) or with control solution (C) for 30 days. Seven days after the beginning of the treatment, the animals received identical inoculum concentrations of M. bovis. Hamsters treated with the higher (2.0 and 3.0 mg kg-1 day-1) doses of diazepam exhibited: 1) increased granuloma areas in the liver (C = 1.81 + or - 1.39, E2 = 10.29 + or - 4.64 and E3 = 15.80 + or - 4.82) and lung (C = 0.54 + or - 0.55, E2 = 6.28 + or - 3.85 and E3 = 6.31 + + or - 3.56) and 2) increased scores of M. bovis colony-forming units isolated from liver (C = 2.0, E2 = 3.0 and E3 = 3.5), lung (C = 1.0, E2 = 3.0 and E3 = 3.5) and spleen (C = 1.0, E2 = 2.5 and E3 = 4.0). These effects were dose dependent, and were not detected or were less severe in animals treated with the lowest (1.0 mg/kg) dose of diazepam as well as in those of the control group. Furthermore, diazepam treatment (3.0 mg kg-1 day-1 for 30 days) increased (E3 = 71.32 + or - 2.99; N = 10) the serum levels of cortisol compared to control hamsters (C = 22.61 + or - 2.75; N = 10). The present data, that demonstrate an impaired defense against M. bovis in adult hamsters treated with diazepam, were tentatively explained on the basis of a direct and/or indirect action of diazepam on the cytokine network. The effects may be related to stimulation of peripheral benzodiazepine receptor binding sites (PBR) by macrophages and/or lymphocytes, or they may be mediated by PBR stimulation of the adrenals

Avaliaçäo quantitativa da concentraçäo de micobactérias em órgäos e humores de hamster experimentalmente infectados com Mycobacterium bovis, estirpe AN 5 / Quantitative evaluation of mycobacterial concentration in organs and fluids of experimentally infected hamster with Mycobacterium bovis, strain AN 5

Descreve-se a tuberculose desencadeada pela inoculaçäo intraperitoneal de 1,0 mg de Mycobacterium bovis (cepa AN 5) em hamsters através de exames bacteriológicos e histopatológicos, realizados 1, 15, 29 e 45 dias após a infecçäo. A bactéria se multiplicou no baço, rins, fígado e pulmöes dos animais infectados, induzindo a formaçäo de lesöes granulomatosas. Um dia após a infecçäo o baço albergou um maior número de bactérias e 45 dias após a infecçäo houve diferença nas concentraçöes bacterianas apenas entre baço e rim, com um maior número de bactérias para o baço. Foram observadas duas fases de bacteremia e a micobactéria foi cultivada a partir do líquido peritoneal em todas as etapas de experimento. Os animais perderam peso em decorrência da doença e morreram em média 50,6 dias após a infecçäo

Arbovirus studies in southwestern Venezuela during 1973-1981. II. Isolations and further studies of Venezuelan and eastern equine encephalitis, Una, Itaqui, and Moju viruses.

Increasing utilization of arable land in southwestern Venezuela has led to a potential increase in human exposure to arbovirus infections. Since previous studies in the Catatumbo region of this area documented the presence of eastern equine encephalitis (EEE) and Venezuelan equine encephalitis (VEE) viruses, an attempt was made to study the transmission and maintenance of these viruses from 1973 to 1981. Isolations of EEE, VEE ID strains, Una, Itaqui , and Moju viruses were repeatedly obtained from mosquitoes, mostly Culex ( Melanoconion ) spp. and sentinel hamsters. The results indicate that these viruses constitute a potential hazard to public health in the area. Further, the strategic location of the Catatumbo region, between enzootic tropical foci of arboviruses, may provide circumstances and conditions for study of both enzootic maintenance and movement of these viruses.

[Features of infection in hamsters by Junín virus]. / Características de la infección del criceto por virus Junín.

Suckling hamsters infected with three low passaged strains of Junin virus by intracerebral route developed a non differentiated illness with about 7 log of virus replication in the brain after which they either died between 6 and 19 days post inoculation or developed CF antibodies. Even in suckling hamsters and suckling mice gave similar results for viral titration, isolation attempts were less successful in hamsters. The young hamster response was characterized by a severe neurological disease and subsequent recovery with the development of CF antibodies 3). Except for isolation efficiency, the results obtained with Junin virus are similar to those reported for other arenavirus.

Studies of arboviruses in Southwestern Venezuela: I. Isolations of Venezuelan and Eastern Equine Encephalitis viruses from sentinel hamsters in the Catatumbo region.

The purpose of this report is to describe isolations of Venezuelan (VEE) and Eastern (EEE) Equine Encephalitis virus made in the lowland moist tropical forest of the Catatumbo region on the southwestern part of the State of Zulia, Venezuela. We have isolated four strains of EEEV from sentinel hamsters exposed at Caño Mocho and Madre Vieja sites in 1973 and 1974, and three strains of EEEV in Hacienda (Hda.) Las Nubes in 1975. Both viruses were recovered during silent interepidemic periods and we believe these viruses are maintained in this region in sylvatic conditions. The recovered virus strains were detected within 24 to 48 hours, both in SMB and Vero Cell monolayer cultures and the sentinel hamsters yielded virus infectivities up to 10(4) PFU ml. Our VEEV isolate (IVIC PAn 23645-47), recovered during the silent interepizodemic period had an elution profile on the hydroxylapatite column corresponding to that of a I-D (VEEV-3880) or a I-E (VEEV-63A216) 'enzootic' subtype. However, considering other in vitro criteria (KHI; HA pH 5.8-6.0; small plaque size in Vero monolayers with suitable overlay media), this later and other previous isolates had some very distinct properties of the 'epizootic' strains. Thus, the evidence suggests that in Venezuela the VEEV cycle in nature is maintained either by the so called 'enzootic' and/or 'epizootic' virus types, or the virus population of the isolates have particular in vitro properties which do not correlate to the virulence markers in vivo. We consider this important question must be further clarified, and in addition, the isolation of three strains of EEEV are reported; this is the first report of the presence of this virus in Venezuela. Although the EEEV isolates may be of the South American type, they must be considered as potentially dangerous in the case of outbreaks.

Search for epizootic-like Venezuelan encephalitis virus at enzootic habitats in Guatemala during 1969-1971.

Seventy-four strains of Venezuelan encephalitis (VE) virus recovered from sentinel hamsters or mosquitoes at enzootic habitats in Guatemala in the two years following the 1969 epidemic-equine epizootic were examined for ability to produce small plaques in Vero African green monkey kidney cell cultures, like isolates obtained during the epizootic. (a) One strain recovered from a sentinel hamster in late October 1969 at an enzootic habitat near the epicenter of the hemagglutination-inhibition (HI) and equine-virulence properties like epizootic virus; this strain retained its small plaque characteristic after inoculation and recovery from bloods of three horses. (b) None of the other 73 strains produced uniformly small plaques, but 31 formed a few small plaques among large ones. Virions from small plaques of five strains were cloned twice in Vero cell cultures. Four clones produced uniformly small plaques after one more passage in Vero cells; three had hemagglutination-pH properties compatible with epizootic virus or intermediate between epizootic and enzootic virus, but HI tests with these three hemagglutinins or with antibody to the fourth cloned strain showed them to be like Central American enzootic virus. One of three cloned strains tested in horses produced encephalitis and death in one of four horses; another strain produced encephalitis with recovery in one of two horses. (c) Thus these small Vero plaque clones resembled Central American enzootic strains of VE virus in HI and equine-virulence tests, and the small Vero plaque characteristic was not a satisfactory marker for consistently isolating equine-virulent, epizootic VE virions. Nevertheless, this technic led to recognition of one epizootic strain isolated at an enzootic habitat in Guatemala at the end of 1969 outbreak. Whether this strain was there before the outbreak or subsequently penetrated the habitat is uncertain. During the next two years, this strain did not become dominant in that enzootic focus.

Detection of Venezuelan equine encephalitis virus in rural communities of Southern Florida by exposure of sentinel hamsters.

Venezuelan equine encephalitis (VEE) virus strains were recovered from sentinel hamsters exposed in close proximity to homes in rural South Florida. Sentinel hamster surveillance methods over extended periods offer one effective way of uncovering VEE virus activity in relation to human habitation.

Rodent control programmes in areas affected by Bolivian haemorrhagic fever.

Bolivian haemorrhagic fever (BHF) caused by Machupo virus is acquired by contact with the excretions and secretions of Calomys callosus, an indigenous cricetine rodent which is preadapted to peridomestic habitats. It competes successfully with Mus musculus, but not with Rattus rattus. A successful disease control programme has functioned in Beni Department since 1964. It is based on trapping surveys and the detection of splenomegaly in Calomys rodents as an index of chronic virus infection. Mass trapping and poisoning are used initially, and regular trapping is employed to control Calomys populations in towns where disease has occurred. More than 1000 cases of BHF were recorded from 1960-1964, but less than 200 in the past 10 years. The cost of this programme is approximately $30 000 annually.