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Associations between leptospiral infection and seropositivity in rodents and environmental characteristics in Argentina.

Our objective was to look for associations between leptospiral infection in rodents and selected environmental and rodent characteristics in Santa Fe, Argentina. Rodents (n = 214) were trapped alive from January 1998 to December 1999 in three environmental settings. Kidneys from 118 rodents were cultured and serum samples from 201 were processed by enzyme-linked immunosorbent assay (ELISA). Logistic regression was performed with ELISA seropositivity as the dependent variable and rodent characteristics were offered as independent variables. Overall prevalence of positive ELISA reactions was 42% (84/201). In urban areas, leptospiral isolations belonged to the Ballum serogroup; in natural corridors, they belonged to the Icterohaemorragiae serogroup. M. musculus (house mouse) was the most-frequently captured species and the predominant one in urban areas. Most isolates and seropositivity results were obtained on this species. Adults and subadults had higher seroprevalences than juvenile rodents. Oligoryzomys flavescens had higher seroprevalence than Akodon azarae, Mus musculus, Rattus rattus and Rattus norvegicus.

Animals↗

Leptospirosis as an animal and public health problem in Latin America and the Caribbean area.

As a major livestock disease, leptospirosis cuts significantly into the world's supply of animal protein through its consequences of abortion and calf mortality, retarded growth, and reduced milk production. Evidently, such losses have severe economic implications as well. The disease's impact has been somewhat diminished in the developed countries thanks to mechanization of rural activities, improved personal and environmental hygiene, and broad application of preventive measures, including the administration of bacterins, in domestic animals. Still, even in these circumstances it remains a serious problem. In the developing countries, in which large percentages of the population live in close contact with animals, the spread of infection is much greater. Moreover, the ecology of tropical and subtropical America--with its abundant rainfall, natural water courses, and high temperatures--is particularly favorable for the transmission of leptospires. In such conditions the human disease is a growing threat as well. The spread of leptospirosis, both in animals and man, is facilitated by the fact that inefection from many of the serotypes can exist without presenting serious clinical manifestations. Hence the importance of laboratory diagnosis. Systematic studies over time of serotype distribution, coupled with wide and improved surveillance, will be a key factor in bringing the disease under control. According to the limited seroepidemiologic data available, 3 to 18 per cent of the general human population, 2 to 32 per cent of those belonging to exposed occupational groups, and 10 to 50 per cent of patients with fever of unknown origin have leptospiral antibody. The most predominant serogroups are Icterohaemorrhagiae, Canicola, Pomona, Hebdomadis, and Bataviae. So far more than 50 serotypes, from 15 serogroups, have been isolated from man and animals in Latin America and the Caribbean area.

Animals↗

[Evidence of dog as a reservoir for human leptospirosis: a serovar isolation, molecular characterization and its use in a serological survey].

Canine leptospirosis has been known as Stuttgart disease since 1898, and dogs are considered to be the second principal source of infection in man. The isolation of a pathogenic serovar from dog urine that was diagnosed clinically and laboratorial as having leptospirosis and its utilization to test serological samples of human and canine cases of leptospirosis, has demonstrated its importance to the ecosystem of the southern region of Brazil. The results of the serological microscopic agglutination test indicated that 100% of human serum samples from 12 patients from the serum bank of 2001 at the Center for Control of Zoonoses, that had titers between 25 and 3,200 with the canicola serovar, and 72% of 105 canine serum samples from the same serum bank, also reacted with the new isolate. The mean and median titers of the human serum samples tested with the battery of antigens recommended by WHO was 630 and 100 respectively, and when tested with the isolate it was 1,823 and 400. In the dog sera, the values were respectively 347 and 100 with the battery, and 1,088 and 200 with the isolate.

Agglutination Tests↗

Leptospiral antibodies in patients from a Barbadian general practice.

Sera from 1,419 patients who attended a Barbadian general practice for a variety of complaints between 1 April 1984 and 30 April 1988 were examined for leptospiral agglutinins by the microscopic agglutination test (MAT). Sera from the 42 patients with pyrexia of unknown origin, jaundice or kidney involvement, and the 26 patients with titres greater than or equal to 1:400 in the MAT, were also examined by the ELISA for IgM and IgG antibodies. Current or recent mild leptospiral infection was diagnosed in four of the patients, but some mild cases may have been missed. Two further cases of leptospirosis were recorded after the study period ended; both were very ill but responded well to doxycycline therapy. Altogether 177 of the 1,419 patients (12.5%) had agglutination titres greater than or equal to 1:50. These were 104/739 males (14%) and 73/680 females (11%). Seropositivity tended to increase with age (p less than 0.01), and the highest rates were in agricultural workers (35%), labourers (24%) and non-manual outdoor workers (19%). The difference in seropositivity between the main occupational groups was highly significant (p less than 0.001). The serogroups most commonly recorded among the seropositive patients were Autumnalis (31%), Panama (26%), Australis (24%) and Pyrogenes (20%). Autumnalis predominated in each of the main occupational groups except indoor non-manual workers where Panama, Pyrogenes and Australis occurred more frequently. 95% of the positive titres ranged between 1:50 and 1:400. Titres tended to increase with age, but there was no obvious association between higher titres and particular occupations.(ABSTRACT TRUNCATED AT 250 WORDS)

Agglutination Tests↗

Leptospiral antigens in the liver of experimentally infected guinea pig and their relation to the morphogenesis of liver damage.

In order to investigate the morphogenes of experimental leptospirosis by morphologic and immunohistologic methods, 24 guinea-pigs were inoculated intraperitoneally with L. interrogans serogroup Icterohaemorrhagiae. They were divided in 6 groups, sacrificed from the 1st to the 6th day of infection. Semiquantitative analyses of histopathological liver lesions were performed in 1 micron sections of tissue embedded in glycol-methacrylate. The distribution of leptospiral antigen (L. Ag) and its glycolipoprotein (GLP) was demonstrated by peroxidase-antiperoxidase on paraffin embedded tissue. Significant lesions appeared at the 4th day of infection, progressing to a peak on the 6th day. Inflammation was associated with injury of the portal triad. Liver cells showed either swelling or acidophilic degeneration and necrosis, together with loss of cell cohesion, leading to disarray of liver cell plates. Mitochondria were found progressively enlarged and irregularly distributed. L. Ag expression was parallel to the morphological changes. Portal distribution was significant at the 4th day and on later stages centrilobular localization became predominant. Spiral forms suggestive of intact leptospires were initially found but, chiefly at the 6th day, L. Ag was seen in granules, probably resulting from phagocytosis. GLP staining was similar to granular L. Ag in morphology, and distribution. Cytokeratin condensation was seen in liver cells with acidophilic necrosis and was marked in areas of disorganization of cell plates. Our findings lead us to hypothesize a direct leptospiral cytotoxic effect on endothelial and on liver-cell membranes. At first, leptospires themselves would induce subcellular changes acting mainly on membrane permeability. Afterwards, their granular forms, including GLP, would act as adjuvant factors. These findings demonstrate that the disarray of liver cell plates at the late phase of the disease is genuine.

Animals↗

Screening of patients with acute febrile illness for leptospirosis using clinical criteria and serology.

BACKGROUND: Leptospirosis is one of the common zoonoses but, in most instances, the infection goes unnoticed. Rapid diagnostic modalities are needed to diagnose the disease in the early stages. We assessed the usefulness of clinical criteria and compared these with enzyme-linked immunosorbentassay (ELISA) for the early detection of leptospirosis. METHODS: One hundred patients with a febrile illness for > 7 days were screened by Faine criteria and their sera were subjected to both IgM and IgG ELISA using a commercially available kit (Institut Virion Serion GmbH, Warburg, Germany). RESULTS: Twenty-six patients satisfied the clinical criteria for leptospirosis and 8 of them tested positive for IgM antibodies while 1 patient who was clinically negative tested positive by serology. Thus, Faine criteria had a sensitivity of 88.9%, specificity of 80.2%, positive predictive value of 30.8% and a negative predictive value of 98.6%. Paired serum samples were obtained from 70 patients but the IgG levels of only 2 showed a 4-fold rise. CONCLUSION: Faine criteria has moderate sensitivity and specificity but a high negative predictive value in comparison with IgM ELISA. The high negative predictive value may help to screen patients with acute febrile illness for leptospirosis during the early phase of the disease.

Acute Disease↗

Detection of leptospiral antigen (L. interrogans serovar copenhageni serogroup Icterohaemorrhagiae) by immunoelectron microscopy in the liver and kidney of experimentally infected guinea-pigs.

Guinea-pigs were experimentally infected with L. interrogans serovar copenhageni serogroup Icterohaemorrhagiae and their liver and kidney were studied by immunoelectron microscopy using the post embedding indirect immunogold labelling technique. Primary antibody was a purified rabbit anti-serum produced against the same leptospiral strain used in the inoculum. Gold-labelled leptospiral antigen (LAg) was found close to cell membranes of hepatocytes, kidney tubular cells and endothelial cells of the interstitial capillaries of the kidney. Afterwards it was internalized by hepatic and tubular cells, and eventually found in lysosomes. Phagolysosomes of Kupffer cells were also found to contain remnants of degraded leptospires and gold-labelled LAg. Gold-labelled intact leptospires were detected at the enlarged intercellular spaces between hepatocytes at the areas of hepatic cell plate disarray, showing the potential for leptospiral migration during the septicaemic phase of the disease potentially contributing to the pathogenesis of the lesions. The affinity of leptospiral antigenic material for cell membranes suggests an initial interaction with cell surface proteins followed by its internalization and cell damage. The nature of antigenic material detected, however, remains undefined; it may be a toxin, an enzyme or any other factor/s involved in leptospiral virulence.

Animals↗

Report of three children with leptospirosis in rural area of the east of Turkey.

Leptospirosis is a systemic infection usually producing fever with hepatorenal involvement, meningoencephalitis, and hemorrhage. In this article, we present three children between 10 and 13 years of age with leptospirosis. The purpose of this paper is to emphasize that leptospirosis is a problem in our country with farmers/cattle and that leptospirosis should be considered in certain ill children. The main symptoms were headache, fever, fatigue, abdominal pain and unconsciousness. Two patients had hepatic and renal involvement. The other had hepatic, pulmonary and probably pericardial involvement. In all children spirochetes were demonstrated in blood and urine smears by dark-field microscopy and they were also isolated from urine and blood cultures by using Flecher medium. All patients were treated with penicillin; however, one subsequently required additional antibiotics due to Klebsiella pneumoniae septicemia. While one patient was discharged in a good health, the others were taken to their home by parents without completing treatment. In conclusion, we would like to emphasize that leptospirosis is still a public health problem in our region (Eastern Turkey) in where the majority of population are farmers and raise domestic animals such as cattle in rural areas. Additionally, leptospirosis should be considered in children admitted with headache, unconsciousness, fever and abdominal pain.

Adolescent↗

Experimental leptospiral infections in pregnant cattle with organisms of the Hebdomadis serogroup.

The clinical and pathologic implications of leptospirosis caused by organisms of the Hebdomadis serogroup were tested on 8 pregnant cows. The animals were inoculated after their 5th month of pregnancy, 2 with serovar szwajizak and 6 with serovar hardjo. During the acute phase of the infection, the cows had mild signs of clinical illness; however, mastitis was observed in all cows that calved. One cow aborted, and 2 delivered premature and weak calves. By bacteriologic cultural examination, a short leptospiremic phase was observed. Leptospires were isolated from milk of 5 cows during early phases of mastitis; these isolations were obtained only when using solid media. Leptospiruria was determined by culture and direct examination. Only hardjo-inoculated cows had detectable urinary shedding which was observed to extend past 450 days in 1 cow. Leptospires were isolated from tissues of all cows and from 2 calves at necropsy. Interstitial nephritis was observed in all cows necropsied. Histologically, leptospires were demonstrated in silver-stained sections of kidney, liver, and lung of most calves. Leptospires were also observed in sections of kidney of all cows necropsied and in the cotyledons, placenta, and liver of some. Agglutinating titers were detected in the serum of cows as early as 4 days after inoculation and reached maximum titer (81,920) after 20 days. Significant serum titers persisted for the duration of the experiment only in hardjo-inoculated cows. At birth, all calves had no detectable agglutinins, but seroconverted within 24 hours; the serum titers were similar to those found in the dam's milk.

Animals↗

Leptospires in the whistling frog (Eleutherodactylus johnstonei) on Barbados.

Two groups of whistling frogs (Eleutherodactylus johnstonei) comprising 99 and 117 animals were examined for leptospiral infection. Group I animals were caught in 14 areas of Barbados, and Group II animals in seven areas of suburban Bridgetown. Leptospires were isolated from the kidneys or body fluid of six frogs in Group I and the kidneys of 3 frogs in Group II. Two of the Group I isolates died out; the others were identified as bajan (a new serovar in the Australis serogroup) (6) and bim (Autumnalis) (1). The macerated body tissues and fluid of Group I frogs were put into phosphate buffered saline and examined by the microscopic agglutination test using 22 antigens. The results were all negative. For the Group II frogs the methodology was altered; blood was collected onto filter paper discs and allowed to dry out before being agitated in PBS and examined by the MAT. 15/117 (12.8%) animals were positive at greater than or equal to 1:100 and 19 (16.2%) at greater than or equal to 1:50. The geometric mean titre was 179. Seventeen of the sera reacted predominantly to antigens in the Australis serogroup, and two to Pyrogenes on its own. The serological results reflected the identity of the isolates. Serovars of Australis are not known to cause illness on Barbados, but bim is the commonest cause of severe leptospirosis on the island.

Animals↗

Leptospiral infection in school-children from Trinidad and Barbados.

A serological survey for leptospiral agglutinins was undertaken between 1980 and 1983 in over 500 Barbadian and 500 Trinidadian school-children aged 7-14 years. The children were selected randomly from urban and rural schools, and examined three times at approximately annual intervals. A total of 12.5% of the Barbadian children and 9.5% of the Trinidadian children were seropositive at a titre of 50 using the microscopic agglutination test. On both islands, seroprevalence was higher in males than females, the difference being significant in rural schools. There was no evidence of a difference in prevalence between urban and rural schools, or between junior and secondary age-ranges. Analysis of the association of serology with socio-economic and behavioural factors showed a significant association in Trinidad with father's occupation, but most other variables on both islands showed only weak non-significant associations. Fourteen children in Trinidad and three in Barbados seroconverted. Seroconversion in Trinidad occurred at a rate of 1.6% per annum and was significantly associated with livestock contact and with absence of a tapped water supply. In Trinidad, Autumnalis was the most commonly recorded serogroup, but this accounted for less than a quarter of seropositives. In Barbados, Panama accounted for over half the seropositives and was about four times more common than the next most common serogroup, Autumnalis. In Barbados, 39 persons aged 19 or less were hospitalized with leptospirosis between November 1979 and December 1986. Average annual incidence rates were 2.2, 4.9 and 13.3 per 100,000 in the 5-9, 10-14 and 15-19 age-groups, respectively.

Adolescent↗

Macroscopic agglutination test for rapid diagnosis of human leptospirosis.

A commercially available slide agglutination test (SAT) for the diagnosis of human leptospirosis was evaluated by comparing it to an immunoglobulin M (IgM) enzyme-linked immunosorbent assay (ELISA) and to the microscopic agglutination test (MAT). For all 108 patients, leptospirosis was diagnosed on the basis of a fourfold or greater increase in titer by MAT (seroconversion), and all but 1 of 245 controls were MAT negative (titers, <1:100). Both SAT and the IgM ELISA failed to detect one case of infection (sensitivity, 99%). Only 3 of 145 blood donors and none of the 100 patients with other illnesses were SAT positive (specificity, 99%). The overall results were similar for the three tests; however, SAT and ELISA were statistically more sensitive as initial screening tests. For 22% of the patients, the diagnosis of leptospirosis was made earlier by SAT than by MAT. SAT detected 27 (44%) of 62 MAT-negative patients with the first serum sample. ELISA and SAT had very similar results. Follow-up of patients for 1 year after the onset of symptoms showed a decreasing rate of positivity by SAT from the third month on. The rate of positivity by ELISA decreased more slowly, to about 67% by the end of the study. By MAT all patients were persistently reactive. SAT and ELISA seem to be convenient methods for the rapid and early screening for leptospirosis and could replace the less sensitive MAT. ELISA gives less subjective results than SAT and provides information on IgM kinetics, but it can be performed only by the more sophisticated laboratories. SAT is inexpensive, can be performed more quickly and more easily than ELISA, and could be used by the less well equipped laboratories.

Adolescent↗

Leptospiral infection among primitive tribes of Andaman and Nicobar Islands.

The Andaman islands were known to be endemic for leptospirosis during the early part of the century. Later, for about six decades no information about the status of the disease in these islands was available. In the late 1980s leptospirosis reappeared among the settler population and several outbreaks have been reported with high case fatality rates. Besides settlers, these islands are the home of six primitive tribes of which two are still hostile. These tribes have ample exposure to environment conducive for transmission of leptospirosis. Since no information about the level of endemicity of the disease among the tribes is available, a seroprevalence study was carried out among all the accessible tribes of the islands. A total of 1557 serum samples from four of the tribes were collected and examined for presence of antileptospiral antibodies using Microscopic Agglutination Test (MAT) employing 10 serogroups as antigens. An overall seropositivity rate of 191% was observed with the highest rate of 53.5% among the Shompens. The seropositivity rates in the other tribes were 16.4% among Nicobarese, 222% among the Onges and 14.8% among the Great Andamanese. All of the tribes except the Onges showed a similar pattern of change in the seroprevalence rates with age. The prevalence rates were rising from low values among children to reach a peak in those aged 2140 years and then declined. Among Onges the seroprevalence rates continued to rise beyond 40 years. In all the tribes, seroprevalence rates were found to be significantly higher among the males. The commonest serogroups encountered were Australis followed by Grippotyphosa, Icterohaemorrhagiae, Pomona and Canicola.

Adolescent↗

Infectious disease and the decline of Steller sea lions (Eumetopias jubatus) in Alaska, USA: insights from serologic data.

Serologic data were examined to determine whether infectious disease may have played a role in the decline of Steller sea lions (Eumetopias jubatus) in the Gulf of Alaska and Aleutian Islands, USA. Available published data, unpublished data, and recent collections (1997-2000) were compared and reviewed. Data were stratified by geography to compare the declining western Alaskan population in the Aleutian Islands through eastern Prince William Sound to the increasing population in southeastern Alaska. Prevalences of antibodies from the 1970s to the early 1990s were noted for Leptospira interrogans, Chlamydophila psittaci, Brucella spp., phocid herpesvirus-1, and calciviruses. Serum samples collected from 1997-2000 were tested for antibodies to these agents as well as to marine mammal morbilliviruses, canine parvovirus, and canine adenovirus-1 and -2. Conclusions could not be drawn about changes in antibody prevalence to these agents during the decline of Steller sea lions, however, because data were incomplete or not comparable as a result of inconsistencies in testing techniques. Despite these shortcomings, results provided no convincing evidence of significant exposure of Steller sea lions to morbilliviruses, Brucella spp., canine parvovirus, or L. interrogans. Steller sea lions have been exposed to phocid herpesviruses, caliciviruses, canine adenovirus, and C. psittaci or to cross-reactive organisms in regions of both increasing and decreasing sea lion abundance. Based on similar antibody prevalence estimates from the increasing and decreasing populations, these agents are unlikely to have been the primary cause of the population decline. They may have contributed to the decline or impeded population recovery, however, because of undetected mortality and morbidity or reductions of fecundity and body condition in animals under other stresses. Systematic monitoring for disease agents and their effects is needed to determine whether infectious disease currently plays a role in the decline and lack of recovery of Steller sea lions.

Alaska↗

[Screening for antibodies against zoonotic agents among employees of the Zoological Garden of Vienna, Schönbrunn, Austria].

The aim of this study was to investigate the prevalence of antibodies against zoonotic agents in employees of the zoological garden of Vienna, Schönbrunn, Austria. Sixty out of 120 employees participated in the study. In 97% of them antibodies to at least one zoonotic agent were identified. Only two participants were free of antibodies to the zoonotic agents tested. The following seroprevalences (in brackets) were obtained: Viral zoonotic (and potentially zoonotic) agents: Influenzavirus A/H1N1 (58%), Influenzavirus A/H3N2 (85%), Lymphocytic choriomeningitis virus (13%), Encephalomyocarditis virus (5%), Orthopox- (Cowpox-) virus and Hantavirus type Puumala (3%). Hantavirus type Hantaan and Borna disease virus (all negative). Bacterial zoonotic agents: Bartonella henselae (65 %), Borrelia burgdorferi (10%), Leptospira interrogans serovar copenhageni and serovar icterohaemorrhagiae as well as Chlamydophila psittoci (2% each). Brucella spp., Coxiella bumetii, and Francisella tularensis (all negative). Parasitic zoonotic agents: Toxoplasma gondii (53%), Toxocara spp. (21%), Capillaria hepatica (2%), Fasciola hepatica, Schistosoma mansoni, E. multilocularis, and E. granulosus (all negative). The remarkably high seroprevalence to the causative agent of cat scratch disease, Bartonella henselae, is probably due to the private contact of the employees to cats. Regarding viral zoonotic agents it has to be mentioned that Influenzavirus vaccination and/or human-to-human transmission of especially A/H3N2 Influenzaviruses probably attributed significantly to the very high seroprevalence to both Influenzavirus types A/H1N1 and A/H3N2. When investigating parasitic zoonotic agents, high prevalence rates were found against Toxoplasma gondii and Toxocara spp., however, it was not possible to establish a causal link between seropositivity and the professional activity in the zoo. Interestingly, in the case of antibodies to T. gondii, the typical correlation with age was not found in this study, while in the case of the Toxocara spp. positive subjects a correlation was identified with both age and duration of employment in the zoo. Regarding the later two zoonotic parasites, employees of the zoological garden showed significantly higher seroprevalences than the average Austrian population. Antibodies to Capillaria hepatica, a hepatic-parasite in rodents which is diagnosed in humans rarely, were identified in one employee and another one showed a questionable positive result. Further investigations did not exhibit clinical infestations with the parasite in these two individuals so far.

Adolescent↗

Exposure to feline and canine pathogens in bobcats and gray foxes in urban and rural zones of a national park in California.

Exposure of bobcats (Lynx rufus) and gray foxes (Urocyon cinereoargenteus) to a range of common canine and feline pathogens was assessed in urban and rural zones of Golden Gate National Recreation Area, a National Park in the San Francisco Bay Area, (California, USA) from 1992 to 1995. Testing included serology for canine distemper virus, canine parvovirus (CPV), canine adenovirus, Leptospira interrogans, feline calicivirus (FCV), feline panleukopenia virus, feline herpesvirus, feline enteric coronavirus (FECV), feline immunodeficiency virus, feline leukemia virus, Toxoplasma gondii, and Bartonella henselae. Testing was also performed for Dirofilaria immitis. Significantly more gray foxes were seropositive for CPV in the urban zone than in the rural zone. In addition, radio-tracking of gray foxes in the rural zone indicated that all three of the rural CPV-seropositive foxes had traveled into adjoining small towns, whereas only one of the 11 seronegative animals had done so. Significantly more bobcats were seropositive for FCV in the rural zone than in the urban zone. Individual bobcats with positive FCV antibody titers had patterns of movement that intercepted park inholdings where domestic cats lived. Bobcat samples were seronegative for all five of the other viral feline pathogens, with the exception of a FECV-seropositive bobcat. High seroprevalence was detected for B. henselae and T. gondii in both zones. Variation in the seroprevalence for different pathogens might be related to differences in the exposure of bobcats and foxes to domestic animals: in the urban zone, gray foxes were located in residential areas outside the park, whereas bobcats were not. Although for most of the pathogens examined there was no relationship between urbanization and exposure, our results for CPV in foxes and FCV in bobcats indicated that proximity to urban areas or contact with humans can increase the risk of disease exposure for wild carnivore populations. Combining behavioral information from radio-tracking with data on pathogen exposure or disease incidence can provide valuable insights into the ecology of wildlife disease that might be missed with broad-scale, population-level comparisons alone.

Animal Identification Systems↗