Detection of leptospires in tissue using an immunoperoxidase staining procedure

Ellis, T. M.; Robertson, G. M.; Hustas, L.; Kirby, Monica B

doi:10.1111/j.1751-0813.1983.tb02849.x

Cited by 42 publications

(17 citation statements)

References 4 publications

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“…Delays in sample collection could have decreased leptospire isolation because tissue biochemical postmortem changes rapidly reduce the number of viable leptospires (Faine 1982). The association observed in this study between IHC and isolation was similar to that obtained by Ellis et al (1983) studying naturally infected porcine kidneys. Conversely, the association between kidney silver staining and culture was greater in the present study than in those conducted by Thiermann (1977) in rats and Songer et al (1983) in house mice.…”

Section: Resul Resul Resul Resul Results Ts Ts Ts Tssupporting

confidence: 72%

“…Lesões microscópicas consistindo de nefrite mononu- (Babudieri 1958). The presence of leptospires can be detected by culture or stain of renal sections by silver impregnation or immunohistochemical techniques (Thiermann 1977, Ellis et al 1983) and histological alterations can be observed with light microscopy by hematoxylin-eosin stain (Scanziani et al 1989). The isolation of leptospires from kidneys of wild house mice has been reported (Brown et al 1960, Brockie 1977, Songer et al 1983, Vanasco et al 2000.…”

Section: Mus Musculus Mus Musculus) ) ) ) )] Foram Capturados 41mentioning

confidence: 99%

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Comparison of three diagnostic techniques for the detection of leptospires in the kidneys of wild house mice (Mus musculus)

et al. 2004

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Forty-one wild house mice (Mus musculus) were trapped in an urban area, near railways, in Santa Fe city, Argentina. Both kidneys from each mouse were removed for bacteriological and histological examination. One kidney was inoculated into Fletcher semi-solid medium and isolates were serologically typed. The other kidney was microscopically examined after hematoxylin-eosin, silver impregnation and immunohistochemical stains. Leptospires, all of them belonging to the Ballum serogroup, were isolated from 16 (39%) out of 41 samples. The presence of the agent was recorded in 18 (44%) and in 19 (46%) out of 41 silver impregnated and immunohistochemically stained samples respectively. Additionally, leptospires were detected in high number on the apical surface of epithelial cells and in the lumen of medullary tubules and they were less frequently seen on the apical surface of epithelial cells or in the lumen of the cortical tubules, which represents an unusual finding in carrier animals. Microscopic lesions consisting of focal mononuclear interstitial nephritis, glomerular shrinkage and desquamation of tubular epithelial cells were observed in 13 of 19 infected and in 10 of 22 non-infected mice; differences in presence of lesions between infected and non-infected animals were not statistically significant (P=0,14). The three techniques, culture, silver impregnation and immunohistochemistry, had a high agreement (k³0.85) and no significant differences between them were detected (P>0.05). In addition, an unusual location of leptospires in kidneys of carrier animals was reported, but a relationship between lesions and presence of leptospires could not be established.

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Section: Resul Resul Resul Resul Results Ts Ts Ts Tssupporting

confidence: 72%

Section: Mus Musculus Mus Musculus) ) ) ) )] Foram Capturados 41mentioning

confidence: 99%

Comparison of three diagnostic techniques for the detection of leptospires in the kidneys of wild house mice (Mus musculus)

et al. 2004

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“…(1983) 14 que exploraram a expressão de antígenos em tecido de rim de porcos com objetivos de diagnóstico veterinário. Esses autores valorizaram apenas as estruturas antigênicas com formato de leptospiras íntegras e mostraram a alta sensibilidade e especificidade do método quando comparado com o diagnóstico obtido por cultura de leptospiras.…”

Section: Resultsunclassified

Glicolipoproteína de Leptospira interrogans sorogrupo icterohaemorrhagiae: distribuição em fígado e rim de cobaias experimentalmente infectadas

Santos

Sakata

Yasuda

et al. 1989

Rev. Inst. Med. trop. S. Paulo

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RESUMOAcredita-se que as lesões teciduais na leptospirose possam decorrer da ação direta das leptospiras, de toxinas sintetizadas ou liberadas durante sua lise. O presente estudo visou a extração quimica da glicolipoproteína (GLP) da leptospira, a produção de anti-soro anti-GLP e a avaliação de sua distribuição em cortes de fígado e rim de cobaias inoculadas e sacrificadas em estudo seqüencial diário até o 6? dia de infecçâo, correspondente ao pico da doença. Procurou-se também correlacionar a expressão tecidual da GLP com o grau de lesões locais, em busca de novos subsídios para a compreensão da patogenia da leptospirose. A QLP foi detectada em fígado e rim de 2 dentre 6 cobaias no 5? dia e em todas as 6 no 6? dia de infecção, sob a forma de grânulos no citoplasma de macrófagos, livres no interstício ou acolados à membrana de células endoteliais e parenquimatosas, especialmente nas regiões mais lesadas. A cronologia do aparecimento da GLP e sua distribuição sugerem tratar-se de produto de lise de leptospiras fagocitadas por macrófagos e que esta substância, conquanto não comprovada como iniciadora das lesões, associa-se a seu agravamento nas etapas mais avançadas da leptospirose.IMTERMOS. Glicolipoproteína; Leptospirose; Imunohistoquímica. INTRODUÇÃOEstudos clínicos e anátomo-patológicos da leptospirose têm sugerido que as lesões ou as disfunções orgânicas possam ser causadas pela ação direta das leptospiras, por toxinas elaboradas por essas bactérias ou liberadas durante a sua lise 2,4,5,6,9,10,11,23,29. Diversas tentativas de isolamento e caracterização de possíveis fatores tó-xicos, foram realizadas por STALHEIM 24 , MIL-LER e cols.* JACKSON & ZEY 16 , KNIGHT e cols. 17 e FAINE e cols. ,5 . A ação citotóxica do sorotipo pomona sobre cultura âe fibroblastos foi demonstrada por YAM e cols. 28 , sem entretanto caracterizar a natureza química responsá-vel por essa atividade tóxica. Já, VINH e cols. 27 extrairam do sorotipo Copenhagen!, tanto de cepas virulentas quanto de cepas avirulentas, uma glicolipoproteína (GLP) de efeito citotóxico sobre fibroblastos de camundongos e sugeriram ser o componente lipídico dessa GLP o fator responsável por essa toxicidade.(1) Divisão de Patologia, Instituto

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“…However, in the present trial, no cross-reaction was observed in the histological sections used as controls for hyperimmune sera against serovar Hardjo and strain LO14, neither in sections cut of the kidney of the animal positive for strain LO14. Ellis et al (5) did not observe crossreactions in the indirect immunoperoxidase assay using hyperimmune serum against serovars Tarassovi, Hardjo and Pomona in swine kidneys naturally infected with serovar Pomona. The possibility of infection by other serovars than those used in the immunoperoxidase assay cannot be ruled out in the present trial, mainly in the animals that presented positive results in the direct examination of urine.…”

Section: Discussionmentioning

confidence: 99%

“…From these fragments, sections cut (5 μm) thick were obtained and stained by HE (3), and indirect immunoperoxidase (5).…”

Section: Methodsmentioning

confidence: 99%

Leptospires detection in kidney, liver and uterus of cows slaughtered in Paraná State, Brazil

Silva¹,

Freitas²,

Anzai³

et al. 2005

Braz. J. Microbiol.

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The objective of this trial was to detect leptospires in fragments of kidney, liver and uterus of 96 cows with unknown sanitary status, randomly chosen at slaughter in Paraná, Brazil. All 96 urine samples were submitted to direct examination using dark field microscopy. Positive samples in the direct examination and all kidney, liver and uterus fragments were cultured in EMJH modified medium and Tween 80/40/LH. Sections cut of kidney, liver and uterus were stained by Hematoxylin-Eosin (HE) and indirect immunoperoxidase with hyperimmune serum against serovar Hardjo (Hardjoprajitno) and strain Londrina 14 (LO14), from serovar Canicola, which was isolated in the northern region of Paraná state, Brazil, used as primary antibodies. Direct examination detected leptospires in the urine of four animals. All attempts to isolate leptospires from urine and kidney, liver and uterus fragments were negative after 16 weeks of incubation. In the HE stain, focal infiltrate of mononuclear inflammatory cells was observed in the renal interstitial area of most animals. In the indirect immunoperoxidase assay using hyperimmune serum against LO14 strain, the kidney of only one animal presented positive results. All fragments of kidney, liver and uterus tested with hyperimmune serum against serovar Hardjo were negative.

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Detection of leptospires in tissue using an immunoperoxidase staining procedure

Cited by 42 publications

References 4 publications

Comparison of three diagnostic techniques for the detection of leptospires in the kidneys of wild house mice (Mus musculus)

Comparison of three diagnostic techniques for the detection of leptospires in the kidneys of wild house mice (Mus musculus)

Glicolipoproteína de Leptospira interrogans sorogrupo icterohaemorrhagiae: distribuição em fígado e rim de cobaias experimentalmente infectadas

Leptospires detection in kidney, liver and uterus of cows slaughtered in Paraná State, Brazil

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