Leptospira transport medium (LTM): A practical tool for leptospires isolation

Guedes, Israel Barbosa; Rocha, Katarine de Souza; Negrão, Manoel Pierre; Souza, Gisele Oliveira de; Castro, Juliana Fernandes de Paula; Cavalini, Matheus Burilli; Filho, Antônio F. de Souza; Neto, Manoel Soares Damasceno; Aizawa, Juliana; Moraes, Carla Cristina Guimarães de; Heinemann, Marcos Bryan

doi:10.1016/j.mimet.2020.105995

Cited by 7 publications

(4 citation statements)

References 31 publications

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“…Likewise, the recovery of viable leptospires from tissue samples (e.g., kidney) not processed on the same day as collection is unlikely because of tissue autolysis [53]. Consideration should be given to transferring urine and tissue into a transport medium [53,54] or a buffered solution if a culture medium is not readily available. Freezing of tissue may be another option as leptospires have been successfully cultured from frozen pig [55] and hamster [53] kidneys.…”

Section: Culturementioning

confidence: 99%

Role of Diagnostics in Epidemiology, Management, Surveillance, and Control of Leptospirosis

et al. 2022

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A One Health approach to the epidemiology, management, surveillance, and control of leptospirosis relies on accessible and accurate diagnostics that can be applied to humans and companion animals and livestock. Diagnosis should be multifaceted and take into account exposure risk, clinical presentation, and multiple direct and/or indirect diagnostic approaches. Methods of direct detection of Leptospira spp. include culture, histopathology and immunostaining of tissues or clinical specimens, and nucleic acid amplification tests (NAATs). Indirect serologic methods to detect leptospiral antibodies include the microscopic agglutination test (MAT), the enzyme-linked immunosorbent assay (ELISA), and lateral flow methods. Rapid diagnostics that can be applied at the point-of-care; NAAT and lateral flow serologic tests are essential for management of acute infection and control of outbreaks. Culture is essential to an understanding of regional knowledge of circulating strains, and we discuss recent improvements in methods for cultivation, genomic sequencing, and serotyping. We review the limitations of NAATs, MAT, and other diagnostic approaches in the context of our expanding understanding of the diversity of pathogenic Leptospira spp. Novel approaches are needed, such as loop mediated isothermal amplification (LAMP) and clustered regularly interspaced short palindromic repeats (CRISPR)-based approaches to leptospiral nucleic acid detection.

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Section: Culturementioning

confidence: 99%

Role of Diagnostics in Epidemiology, Management, Surveillance, and Control of Leptospirosis

et al. 2022

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“…Isolating bacteria in urine samples is complex, challenging and can take months to obtain a result. According to Guedes et al (2020), isolation is considered a definitive diagnosis for leptospirosis and useful in identifying renal carriers. However, factors such as the short-term viability of Leptospira in urine, their fastidious nature and the lack of standardized protocols make it difficult to obtain successful results.…”

Section: Resultsmentioning

confidence: 99%

Alterações clínico-patológicas, diagnóstico sorológico e molecular em cães com suspeita de leptospirose

et al. 2023

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A leptospirose é uma zoonose de importância mundial que causa mortes tanto em humanos, como em animais, especialmente em cães não vacinados. A infecção em cães ocorre pelo contato com urina ou água contaminada pela bactéria espiroqueta patogênica do gênero Leptospira. Objetivou-se avaliar dois testes diagnósticos na caracterização de cães com leptospirose, e associar com as principais alterações clínicos-patológicas de cães suspeitos. Foram atendidos no Hospital Veterinário da Universidade Federal de Uberlândia no período de setembro de 2019 a setembro de 2020, 24 cães com suspeita clínica de leptospirose canina. Exames complementares como hemograma completo, dosagens séricas de creatinina, ureia, ALT (alanina aminotransferase), FA (fosfatase alcalina) foram solicitados. Para o diagnóstico confirmatório de leptospirose foi realizado teste sorológico pelo teste de aglutinação microscópica (MAT) e também teste molecular pela reação em cadeia da polimerase em tempo real (qPCR). Dos 24 casos de cães suspeitos de leptospirose, seis (25%) foram positivos no MAT, e três (12,5%) no qPCR, sendo 1 (um) (4,17%) positivo nos dois testes diagnósticos avaliados. Quando se associou MAT + qPCR a positividade foi para 8/24 (33,3%). Os sorogrupos reativos predominantes foram Icterohaemorrhagiae (33,33%) e Djasiman (33,33%), seguido de Ballum (16,60%). A sensibilidade e especificidade da qPCR em relação teste padrão ouro (MAT) foi 16,7 % e 88,9%, respectivamente. Houve alterações nos níveis séricos de creatinina, ureia e FA no grupo positivo aos testes diagnósticos (MAT/qPCR), porém somente ureia e FA apresentaram valores elevados no grupo negativo. Quando se comparou os parâmetros bioquímicos no grupo positivo aos testes diagnósticos (MAT/qPCR), e negativo, não se observou nenhuma diferença estatística significativa entre os elementos avaliados (p>0,05). Conclui-se para o diagnóstico de leptospirose canina aguda, o médico veterinário deve associar os achados clínicos-patológicos aos resultados do MAT de forma pareada e o qPCR de urina.

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“…Although culture is commonly used to diagnose bacterial diseases and is considered the gold standard for leptospirosis diagnosis [9,14], it is not recommended for diagnostic purposes. Leptospires are fastidious and very nutrient-demanding bacteria; therefore, their culture takes too long and presents low sensitivity [61]. On the other hand, molecular techniques such as PCR and qPCR present high sensitivity and specificity [62], are fast to conduct [63], and can be performed in various types of clinical samples such as urine, cervical-genital mucus, endometrial biopsy, semen, fetal tissues, and placenta [17,22,45].…”

Section: Molecular Diagnosismentioning

confidence: 99%

Bovine Genital Leptospirosis: An Update of This Important Reproductive Disease

Aymée,

Mendes,

Lilenbaum

2024

Animals

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Bovine leptospirosis is an important disease that affects the reproductive sphere. Due to its high relevance for the bovine production chain in a worldwide scenario, a better understanding of the disease is crucial to reduce its negative impacts. The main agents are strains from the Sejroe serogroup, such as Hardjo and Guaricura, which lead to renal and genital infection. The genital colonization causes a chronic, silent, and subclinical reproductive syndrome, called Bovine Genital Leptospirosis (BGL). Embryonic death, estrus repetition, subfertility, and abortions are the main signs of BGL condition in females. However, although leptospires have been identified in semen, the manifestation of BGL in bulls remains to be clarified. The recommended diagnosis of BGL includes a serologic screening of the herds using the microscopic agglutination test followed by PCR of genital samples (cervicovaginal mucus, uterine fragment, or semen), especially from animals with reproductive failures. After the identification of carriers, control is carried out considering three steps: antimicrobial treatment of the carriers, environmental and reproductive management, and herd vaccination. Systematic testing, quarantine of newly arrived animals, and usage of antimicrobials in semen diluents or embryo culture media are other sanitary approaches that are encouraged to improve the control of the syndrome. Herein we discuss protocols for an efficient diagnosis and preventive procedures of BGL, which are fundamental to reducing the negative impact of the disease on cattle reproduction and its consequent economic hazards.

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Leptospira transport medium (LTM): A practical tool for leptospires isolation

Cited by 7 publications

References 31 publications

Role of Diagnostics in Epidemiology, Management, Surveillance, and Control of Leptospirosis

Role of Diagnostics in Epidemiology, Management, Surveillance, and Control of Leptospirosis

Alterações clínico-patológicas, diagnóstico sorológico e molecular em cães com suspeita de leptospirose

Bovine Genital Leptospirosis: An Update of This Important Reproductive Disease

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