Clinical-pathological alterations, serological and molecular diagnosis in dogs with suspected leptospirosis
DOI:
https://doi.org/10.5433/1679-0359.2023v44n2p823Keywords:
Serum biochemistry, Canine. Leptospira spp., Aclimatização, qPCR, Sensitivity.Abstract
Leptospirosis is a globally important zoonosis that causes death in humans and animals, especially unvaccinated dogs. Dogs are infected through contact with urine or water contaminated by pathogenic spirochaete bacteria of the genus Leptospira. This study aimed to assess the performance of two diagnostic tests in characterizing dogs with leptospirosis and associate it with the main clinical-pathological alterations in dogs with suspected infection. From September 2019 to September 2020, 24 dogs with suspected canine leptospirosis were treated at the Veterinary Hospital of the Federal University of Uberlândia (UFU). Complementary tests such as a complete blood count (CBC) and serum creatinine, ALT (alanine transaminase) and ALP (alkaline phosphatase) levels were prescribed. In order to confirm the leptospirosis diagnosis, serum and molecular tests were performed using the microscopic agglutination test (MAT) and real-time quantitative polymerase chain reaction (qPCR). Of the 24 suspected cases of canine leptospirosis, six (25%) were positive in the MAT and three (12.5%) in the qPCR, with one (4.17%) returning positive results in both diagnostic tests. Positive results were obtained for 8/24 (33.3%) for combined MAT + qPCR. The predominant reactive serogroups were Icterohaemorrhagiae (33.33%) and Djasiman (33.33%), followed by Ballum (16.60%). The sensitivity and specificity of the qPCR in relation to the gold standard test (MAT) were 16.7 and 88.9%, respectively. Changes were observed in serum levels of creatinine, urea and ALP in the group that tested positive (MAT/qPCR); however, only urea and ALP levels were high in the negative group. Comparison of biochemical parameters in the positive (MAT/qPCR) and negative groups showed no statistically significant difference between the elements assessed (p>0.05). It was concluded that in order to diagnose acute canine leptospirosis, veterinarians should combine the clinical-pathological findings with paired sample MAT and qPCR urine test results.
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References
Action software (2020). Estatcamp - consultoria estatística e qualidade. http://www.portalaction.com.br/
Andrade, M. C., Oliveira, L. B., Santos, A. F., Moreira, M. V. L, Pierezan, F., & Ecco, R. (2020). Diagnósticos diferenciais em 83 cães com icterícia. Pesquisa Veterinária Brasileira, 40(6), 451-465. doi: 10.1590/1678-5150-pvb-6482 DOI: https://doi.org/10.1590/1678-5150-pvb-6482
Azócar‐Aedo, L., & Monti, G. (2016). Meta-Analyses of Factors Associated with Leptospirosis in Domestic Dogs. Zoonoses Public Health, 63(4), 328-336. doi: 10.1111/zph.12236 DOI: https://doi.org/10.1111/zph.12236
Chideroli, R. T., Bracarense, A. P. F. R. L., Padovani, L., Martins, L. A., Gonçalves, D. D., & Freitas, J. C. de. (2016). Leptospirose canina associada à insuficiência renal aguda - relato de caso. Revista Brasileira de Medicina Veterinária, 38(Supl. 1), 79-84. https://www.researchgate.net/publication/309852402_Leptospirose_canina_associada_a_insuficiencia_renal_aguda_-Relato_de_caso
Costa, E. R. A., Honorato, R. A., Fiuza, R. F., & Leite, A. K. R. M. (2013). Alterações hematológicas, morfológicas sanguíneas e bioquímicas em um cão com leptospirose. Revista Científica Eletrônica de Medicina Veterinária, 11(21), 1-10. http://faef.revista.inf.br/site/e/medicina-veterinaria-21-edicao-22013.html#tab906
Curtis, K. M., Foster, P. C., Smith, P. S., Monn, M. P., Stillman, B. A., Chandrashekar, R., Lappin, M. R., & Goldstein, R. E. (2015). Performance of a recombinant LipL32 based rapid in-clinic ELISA (SNAP® Lepto) for the detection of antibodies against leptospira in dogs. The International Journal of Applied Research in Veterinary Medicine, 3(3), 182-189. http://www.jarvm.com/
Di Azevedo, M. I. N., Santanna, R., Carvalho-Costa, F. A., & Lilenbaum, W. (2022). The same strain leading to different clinical outcomes: the enigma behind the canine leptospirosis. Microbial Pathogenesis, 165(1), 105500. doi: 10.1016/j.micpath.2022.105500 DOI: https://doi.org/10.1016/j.micpath.2022.105500
Escarrone, A., Baltodano, J. P. R., & Oliveira, M. T. (2020). Leptospirose canina causada pelo sorovar Djasiman no município de Alegrete RS- Relato de caso. Anais do Salão Internacional de Ensino, Pesquisa e Extensão, 8(5), 1.
Ferreira, J. M., Neto, Viana, E. S., & Magalhães, L. M. (1982). Patologia clínica veterinária (2a ed.). Rabelo.
Fraune, C. K., Schweighauser, A., & Francey, T. (2013). Evaluation of the diagnostic value of serologic microagglutination testing and a polymerase chain reaction assay for diagnosis of acute leptospirosis in dogs in a referral center. Journal of the American Veterinary Medical Association, 242(10), 1373-1380.doi: 10.2460/javma.242.10.1373 DOI: https://doi.org/10.2460/javma.242.10.1373
Geisen, V., Stengel, C., Brem, S., Müller, W., Greene, C., & Hartmann, K. (2007). Canine leptospirosis infections - clinical signs and outcome with different suspected Leptospira serogroups (42 cases). The Journal of Small Animal Practice, 48(6), 324-328. doi: 10.1111/j.1748-5827.2007.00324.x DOI: https://doi.org/10.1111/j.1748-5827.2007.00324.x
GraphPad Prism version 8.0.0 for Windows (2022). http://www.graphpad.com
Greene, C. E., Sykes, J. E., Brown, C. A., & Hartmann, K. (2006). Infectious diseases of the dog and cat (3nd ed.). Saunders Elsevier.
Greenlee, J. J., Alt, D. P., Bolin, C. A., Zuerner, R. L., & Andreasen, C. B. (2005). Experimental canine leptospirosis caused by Leptospira interrogans serovars pomona and bratislava. American Journal of Veterinary Research, 66(10), 1816-1822. doi: 10.2460/ajvr.2005.66.1816 DOI: https://doi.org/10.2460/ajvr.2005.66.1816
Guedes, I. B., Rocha, K. S., Negrão, M. P., Souza, G. O., Castro, J. F. P., Cavalini, M. B., Souza, A. F., Fº., Damasceno, M. S., Neto, Aizawa, J., Moraes, C. C. G., & Heinemann, M. B. (2020). Leptospira transport medium (LTM): a practical tool for leptospires isolation. Journal of Microbiological Methods, 175(1), 105995. doi: 10.1016/j.mimet.2020.105995 DOI: https://doi.org/10.1016/j.mimet.2020.105995
Harkin, K. R., Roshto, Y. M., Sullivan, J. T., Purvis, T. J., & Chengappa, M. M. (2003). Comparison of polymerase chain reaction assay, bacteriologic culture, and serologic testing in assessment of prevalence of urinary shedding of leptospires in dogs. Journal of the American Veterinary Medical Association, 222(9), 1230-1233. doi: 10.2460/javma.2003.222.1230 DOI: https://doi.org/10.2460/javma.2003.222.1230
Hartskeerl, R. A., Collares-Pereira, M., & Ellis, W. A. (2011). Emergence, control and re-emerging leptospirosis: dynamics of infection in the changing world. Clinical Microbiology and Infection, 17(4), 494-501. doi: 10.1111/j.1469-0691.2011.03474.x DOI: https://doi.org/10.1111/j.1469-0691.2011.03474.x
IBM SPSS Statistics for Windows, Versão 20.0. (2019). https://www.ibm.com/br-pt/products/spss-statistics
Kaneko, J. J., Harvey, J. W., & Bruss, M. L. (2008). Clinical biochemistry of domestic animals (6nd ed.). Academic Press.
Latosinski, G. S., Fornazari, F., Babboni, S. D., Caffaro, K., Paes, A. C., & Langoni, H. (2018). Serological and molecular detection of Leptospira spp in dogs. Revista da Sociedade Brasileira de Medicina Tropical, 51(3), 364-367. doi: 10.1590/0037-8682-0276-2017 DOI: https://doi.org/10.1590/0037-8682-0276-2017
Levett, P. N. (2001). Leptospirosis. Clinical Microbiology Reviews, 14(2), 296-326. doi: 10.1128/CMR.14.2.296-326.2001 DOI: https://doi.org/10.1128/CMR.14.2.296-326.2001
Major, A., Schweighauser, A., & Francey, T. (2014). Increasing incidence of canine leptospirosis in Switzerland. International Journal of Environmental Research and Public Health, 11(7), 7242-7260. doi: 10.3390/ijerph110707242 DOI: https://doi.org/10.3390/ijerph110707242
Martin, P. L., Stanchi, N. O., Brihuega, B. F., Galli, L., & Arauz, M. S. (2019). Diagnosis of canine leptospirosis: evaluation of two PCR assays in comparison with the microagglutination test. Pesquisa Veterinária Brasileira, 39(4), 255-262. doi: 10.1590/1678-5150-pvb-5868 DOI: https://doi.org/10.1590/1678-5150-pvb-5868
Meira, C. D., Wenceslau, A. A., Carvalho, F. S., Correa, J. M. X., & Albuquerque, G. R. (2011). Detecção molecular de leptospira em amostras de urina de cães infectados naturalmente. Veterinária e Zootecnia, 18(2), 249-254. https://rvz.emnuvens.com.br/rvz/article/view/1116
Meny, P., Menéndez, C., Ashfield, N., Quintero, J., Rios, C., Iglesias, T., Schelotto, F., & Varela, G. (2019). Seroprevalence of leptospirosis in human groups at risk due to environmental, labor or social conditions. Revista Argentina de Microbiología, 51(4), 324-333. doi: 10.1016/j.ram.2019.01.005 DOI: https://doi.org/10.1016/j.ram.2019.01.005
Miotto, B. A., Hora, A. S. da, Taniwaki, S. A., Brandão, P. E., Heinemann, M. B., & Hagiwara, M. K. (2018b). Development and validation of a modified TaqMan based real-time PCR assay targeting the lipl32 gene for detection of pathogenic Leptospira in canine urine samples. Brazilian Journal of Microbiology, 49(3), 584-590. doi: 10.1016/j.bjm.2017.09.004 DOI: https://doi.org/10.1016/j.bjm.2017.09.004
Miotto, B. A., Tozzi, B. F, Penteao, M., Guilloux, A. G. A., Moreno, L. Z., Heinemann, M. B., Moreno, A. M., Lilenbaum, W., & Hagiwara, M. K. (2018a). Diagnosis of acute canine leptospirosis using multiple laboratory tests and characterization of the isolated strains. BMC Veterinary Research, 14(1), 22. doi: 10.1186/s12917-018-1547-4 DOI: https://doi.org/10.1186/s12917-018-1547-4
Mohammed, H., Nozha, C., Hakim, K., & Abdelaziz, F. (2011). Leptospira: morphology, classification and pathogenesis. Journal of Bacteriology & Parasitology, 2(6), 1-5. doi: 10.4172/2155-9597.1000120 DOI: https://doi.org/10.4172/2155-9597.1000120
Oliveira, S. T. (2010). Leptospirose canina: dados clínicos, laboratoriais, e terapêuticos em cães naturalmente infectados. Tese de doutorado, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brasil.
Oliveira, S. T., Messick, J. B., Biondo, A. W., Santos, A. P. dos, Stedile, R., Dalmolin, M. L., Guimaraes, A. M. D., Mohamed, A. S., & Gonzalez, F. H. D. (2012). Exposure to Leptospira spp. in sick dogs, shelter dogs and dogs from an endemic area: points to consider. Acta Scientiae Veterinariae, 40(3), 1-7. https://www.bvs-vet.org.br/vetindex/periodicos/acta-scientiae-veterinariae/
World Health Organization -WHO (2003). Leptospirose humana: orientações para diagnóstico, vigilância e controle. https://apps.who.int/iris/bitstream/handle/10665/42667/WHO_CDS_CSR_EPH_2002.23.pdf?sequence=1&isAllowed=y
Paes, A. C. (2016). Leptospirose canina. In J. Megid, M. G. Ribeiro, & A. C. Paes (Eds.), Doenças infecciosas em animais de produção e de companhia (pp. 356-377). São Paulo.
Picardeau, M. (2013). Diagnosis and epidemiology of leptospirosis. Médecine et Maladies Infectieuses, 43(1), 1-9. doi: 10.1016/j.medmal.2012.11.005 DOI: https://doi.org/10.1016/j.medmal.2012.11.005
Pinna, M. H., Oriá, A. P., Cypriano, G. B., Oliveira, F. S., Almeida, D. S., Pinheiro, A. C. O., Macêdo, L. D., & Rolemberg, D. S. (2010). Leptospirose em cães. PubVet, 4(32), 2. https://ojs.pubvet.com.br/index.php/revista/article/view/2479
Reagan, K. L., & Sykes, J. E. (2019). Diagnosis of canine Leptospirosis. The Veterinary Clinics of North America. Small Animal Practice, 49(4), 719-731. doi: 10.1016/j.cvsm.2019.02.008 DOI: https://doi.org/10.1016/j.cvsm.2019.02.008
Ricardo, T., Previtali, M. A., & Signorini, M. (2020). Meta-analysis of risk factors for canine leptospirosis. Preventive Veterinary Medicine, 181(1), 105037. doi: 10.1016/j.prevetmed.2020.105037 DOI: https://doi.org/10.1016/j.prevetmed.2020.105037
Rissi, D. R., & Brown, C. A. (2014). Diagnostic features in 10 naturally occurring cases of acute fatal canine leptospirosis. Journal of Veterinary Diagnostic Investigation, 26(6), 799-804. doi: 10.1177/1040638714553293 DOI: https://doi.org/10.1177/1040638714553293
Robi, D. T. (2020). Epidemiology and zoonotic implication of leptospirosis in domestic animals in ethiopia. Academic Journal of Animal Diseases, 9(1), 19-32. doi: 10.5829/idosi.ajad.2020.19.32
Rohilla, P., Khurana, R., Kumar, A., Batra, K., & Gupta, R. (2020). Detection of Leptospira in urine of apparently healthy dogs by quantitative polymerase chain reaction in Haryana, India. Veterinary World, 13(11), 2411-2415. doi: 10.14202 / vetworld.2020.2411-2415 DOI: https://doi.org/10.14202/vetworld.2020.2411-2415
Schuller, S., Francey, T., Hartmann, K., Hugonnard, M., Kohn, B., Nally, J. E., & Sykes, J. (2015). European consensus statement on leptospirosis in dogs and cats. The Journal of Small Animal Practice, 56(3), 159-179. doi: 10.1111/jsap.12328 DOI: https://doi.org/10.1111/jsap.12328
Sevá, A. P., Brandão, A. P. D., Godoy, S. N., Souza, G. O., Souza, A. F., Fº., Jimenez-Villegas, T., Heinemann, M. B., & Ferreira, F. (2020). Soroprevalência e incidência de Leptospira spp. em cães domésticos na região Sudeste do Estado de São Paulo, Brasil. Pesquisa Veterinária Brasileira, 40(5), 399-407. doi: 10.1590/1678-5150-pvb-6390 DOI: https://doi.org/10.1590/1678-5150-pvb-6390
Stoddard, R. A., Gee, J. E., Wilkins, P. P., McCaustland, K., & Hoffmaster, A. R. (2009). Detection of pathogenic Leptospira spp. through TaqMan polymerase chain reaction targeting the LipL32 gene. Diagnostic Microbiology and Infectious Disease, 64(3), 247-255. doi: 10.1016/j.diagmicrobio.2009.03.014 DOI: https://doi.org/10.1016/j.diagmicrobio.2009.03.014
Sykes, J., Hartmann, K., Lunn, K., Moore, G., Stoddard, R., & Goldstein, R. (2011). 2010 ACVIM Small animal consensus statement on Leptospirose: diagnosis, epidemiology, treatment, and prevention. Journal of Veterinary Internal Medicine, 25(1), 1-13. doi: 10.1111/j.1939-1676.2010.0654.x DOI: https://doi.org/10.1111/j.1939-1676.2010.0654.x
Thrall, M. A., Weiser, G., Alisson, R. W., & Campbell, T. W. (2015). Hematologia e bioquímica clínica veterinária (2a ed.). Roca.
Troìa, R., Balboni, A., Zamagni, S., Frigo, S., Magna, L., Perissinotto, L., Battilani, M., & Dondi, F. (2018). Prospective evaluation of rapid point-of-care tests for the diagnosis of acute leptospirosis in dogs.Veterinary Journal, 237(1), 37-42. doi: 10.1016/j.tvjl.2018.05.010 DOI: https://doi.org/10.1016/j.tvjl.2018.05.010
Waggoner, J. J., Balassiano I., Mohamed-Hadley, A., Vital-Brazil, J. M., Sahoo, M. K., & Pinsky, B. A. (2015). Reverse-transcriptase PCR detection of Leptospira: absence of agreement with single-specimen microscopic agglutination testing. PLoS One, 10(7), e0132988. doi: 10.1371/journal.pone.0132988 DOI: https://doi.org/10.1371/journal.pone.0132988
Weiss, D. J., & Wardrop, K. J. (2010). Schalm´s veterinary hematology (6nd ed.). Wiley-Backwell.
Zakeri, S., Khorami, N., Ganji, Z. F., Sepahian, N., Malmasi, A. A., Gouya, M. M., & Djadid, N. D. (2010). Leptospira wolffii, a potential new pathogenic Leptospira species detected in human, sheep and dog. Infection, Genetics and Evolution, 10(2), 273-277. doi: 10.1016/j.meegid.2010.01.001 DOI: https://doi.org/10.1016/j.meegid.2010.01.001
Zarantonelli, L., Suanes, A., Meny, P., Buroni, F., Nieves, C., Salaberry, X., Briano, C., Ashfield, N., Silveira, C. S., Dutra, F., Easton, C., Fraga, M., Giannitti, F., Hamond, C., Rioseco, M., Menendez, C., Mortola, A., Picardeau, M., Quintero, J., … Buschiazzo, A. (2018). Isolation of pathogenic Leptospira strains from naturally infected cattle in Uruguay reveals high serovar diversity, and uncovers a relevant risk for human leptospirosis. PLOS Neglected Tropical Diseases, 12(9), 1-22. doi: 10.1371/journal.pntd.0006694 DOI: https://doi.org/10.1371/journal.pntd.0006694
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