Catalase and Glutathione Peroxidase in Dogs Naturally Infected by Leishmania infantum

Belarmino Eugênio Lopes-Neto, Glauco Jonas Lemos Santos, Adam Leal Lima, Maritza Cavalcante Barbosa, Talya Ellen Jesus dos Santos, Daniel Couto Uchoa, Ana Débora Nunes Pinheiro, Romélia Gonçalvez Pinheiro, Diana Célia Sousa Nunes-Pinheiro

Abstract


Background: Canine leishmaniasis (CanL) is caused by an obligatory intracellular parasite of Leishmania genus that
 affects organs and tissues. Several studies evaluate the role of reactive oxygen species (ROS) in the pathogenesis of many diseases. The overproduction of ROS on infectious diseases can induce an imbalance between oxidants and antioxidants at cellular or systemic level. Thus, the aim of this study was to evaluate the activity of antioxidant enzymes in CanL.

Materials, Methods & Results: Females (n = 17) and males (n = 10), at different ages and with different weight, were selected for this study. Dogs were divided into two groups according classical clinical signs and sorological test to CanL. Animals were considered infected based on indirect immunofluorescent assay and ELISA titration ≥ 1:40.  Group B (n = 15) composed by positive dogs to CanL from Zoonosis Control Center of Fortaleza (Ceará, Brazil) and group A (n = 12) was composed by dogs from private kennel that were serologically negative to L. infantum and had absence of clinical signs to CanL. Blood sample were collected for evaluation of hematological and biochemical parameters and glutathione peroxidase (GPx) and catalase (CAT) enzymatic activity. Data were analyzed by Student’s t-test and Pearson correlation coefficient (P < 0.05). Total proteins (TP, mg/dL) and alkaline phosphatase (ALP, U/L) were increased (P < 0.05) on group B (8.2 ± 1.2; 165.4 ± 46.4) when compared to group A (6.5 ± 1.1; 109.1 ± 38.3), respectively. Hemoglobin (Hb; g/dL) and hematocrit (Hct; %) were decreased (P < 0.05) on Group B (14.7 ± 1.8; 48.2 ± 5.7) when compared to group A (16.5 ± 1.3; 52.1 ± 2.4), respectively. Group B presented CAT (U/g Hb) and GPx (mU/mg Hb) lower (189.4 ± 90.4; 3,609.6 ± 1,569.1) than group A (326.6 ± 104.5; 5,055.6 ±1,569.1), respectively (P < 0.001). Positive correlation was observed between RBC and CAT; however, it was not significant.

Discussion: Organisms require a good defense system in order to revert the overproduction of free radicals and consequently the injuries caused by them. This is possible through the production of antioxidant agents, which act on oxidative prevention and on tissue and cellular regeneration, by taking the reduced glutathione (GSH), superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) enzymes in the frontline. Erythrocyte changes promoted by CanL suggest possible correlation between anemia and the appearance of clinical signs, which in many cases is not seen. Erythrocytes contain SOD, CAT and GPx enzymes, thus, changes in these cells will reflect on the activity of these enzymes. In our results only CAT showed positive correlation with erythrocyte count, however it was not significant. GPx activity was lower (P < 0.001) in infected dogs than control group. This result agrees with another study, which showed a decrease in GPx levels in CanL, although it was not significant. However, it was found a positive correlation (P < 0.001) between erythrocytes and GPx activity and between hemoglobin and GPx activity in animals with leishmaniasis. These results suggest that the reduction in detoxification activity can be related to the decrease in erythrocyte count and that the GPx activity depends on the control mechanism of the antioxidant system in CanL. Furthermore, this result could be associated with decrease of blood cell count in animals with CanL, once GPx is an erythrocyte enzyme, which plays an important role in hemoglobin protection against oxidative damage. This study was carried out in naturally infected dogs with L. infantum. In conclusion, CAT and GPx activities are relate to oxidative stress induced by L. infantum infection and can be used as biomarkers on CanL.


Keywords


canine leishmaniasis; antioxidants enzymes; oxidative stress; biomarkers.

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References


Aebi H. 1984. Catalase in vitro. Methods in Enzymology. 105(1): 121-128.

Almeida B.F.M., Narciso L.G., Melo L.M., Preve P.P., Bosco A.M., Lima V.M.F. & Ciarlini P.C. 2013. Leishmaniasis causes oxidative stress and alteration of oxidative metabolism and viability of neutrophils in dogs. The Veterinary Journal. 198: 599-605.

Alvar J., Vélez I.D., Bern C., Herrero M., Desjeux P., Cano J., Jannin J. & Boer M. 2012. Leishmaniasis worldwide and global estimates of its incidence. PLoS One. 7(5): 1-12.

Bae S.Y., Oh H., Rhee S.G. & Yoo Y.D. 2011. Regulation of reactive oxygen species generation in cell signaling. Molecules and Cells. 32(6): 491-509.

Baldissera M. D., Sousa K.C.M., André M.R., Guarda N.S., Moresco R.N., Herrera H.M., Machado R.Z., Jaques J.A.S., Tinucci-Costa M. & Silva A.S. 2015. Nitric oxide, protein oxidation and total antioxidant levels in serum of dogs naturally infected by Ehrlichia canis, Leishmania infantum and Babesia vogeli. Acta Scientiae Veterinariae. 43: 1320.

Barbosa K.B., Costa N.M., Alfenas R.C., de Paula S.O., Minim V.P. & Bressan J. 2010. Estresse oxidativo: conceito, implicações e fatores modulatórios. Revista de Nutrição. 23(4): 629-643.

Bildik A., Kargın F., Seyrek K., Pasa S. & Özensoy S. 2004. Oxidative stress and non-enzymatic antioxidative status in dogs with visceral leishmaniasis. Research in Veterinary Science. 77: 63-66.

Britti D., Sconza S., Morittu V.M., Santori D. & Boari A. 2008. Superoxide dismutase and glutathione peroxidase in the blood of dogs with leishmaniasis. Veterinary Research Communications. 32(Suppl1): s251-s254.

Cardoso L., Schallig H.D.F.H., Cordeiro-da-Silva A., Cabral M., Alunda J.M. & Rodrigues M. 2007. Anti-Leishmania humoral and cellular immune responses in naturally infected symptomatic and asymptomatic dogs. Veterinary Immunology and Immunopathology. 117: 35-41.

Chaudhuri S., Varshney J.P. & Patra R.C. 2008. Erythrocytic antioxidant defense, lipid peroxides level and blood iron, zinc and copper concentrations in dogs naturally infected with Babesia gibsoni. Research in Veterinary Science. 85(5): 120-124.

Circu M.L. & Aw T.Y. 2010. Reactive oxygen species, cellular redox systems, and apoptosis. Free Radical Biology & Medicine. 48(6): 749-762.

Dimri U., Ranjan R., Kumar N., Sharma M.C., Swarup D., Sharma B. & Kataria M. 2008. Changes in oxidative stress indices, zinc and copper concentrations in blood in canine demodicosis. Veterinary Parasitology. 154(1): 98-102.

Dimri U., Singh S.K., Sharma M.C., Behera S.K., Kumar D. & Tiwari P. 2012. Oxidant/antioxidant balance, minerals status and apoptosis in peripheral blood of dogs naturally infected with Dirofilaria immitis. Research in Veterinary Science. 93(1): 296-299.

Freitas J.C.C., Lopes-Neto B.E., Abreu C.R., Coura-Vital W. Braga, S. L. Reis, A.B. & Nunes-Pinheiro D.C.S. 2012. Profile of anti-Leishmania antibodies related to clinical picture in canine visceral leishmaniasis. Research in Veterinary Science. 93(2): 705-709.

Freitas J.C.C., Nunes-Pinheiro D.C.S., Lopes-Neto B.E., Santos G.J., Abreu C.R., Braga R.R., Campos R.M. & Oliveira L.F. 2012. Clinical and laboratory alterations in dogs naturally infected by Leishmania chagasi. Revista da Sociedade. Brasileira de Medicina Tropical. 45(1): 24-29.

Heidarpour M., Soltani S., Mohri M. & Khoshnegah J. 2012. Canine visceral leishmaniasis: relationships between oxidative stress, liver and kidney variables, trace elements, and clinical status. Parasitology Research. 111(4): 14911496.

Jain N.C. 2000. Normal Hematology in Dogs. In: Feldman B.F., Zinkl J.G. & Jain N.C. (Eds). Schalm’s, Veterinary Hematology. 5th edn. Philadelphia: Lippincott Williams & Wilkin, pp.1344-1346.

Karadeniz A., Hanedan B., Cemek M. & Börküm K. 2008. Relationship between canine distemper and oxidative stress in dogs. Revue de Médecine Vétérinaire. 159(8): 462-467.

Kaye P. & Scott P. 2011. Leishmaniasis: complexity at the host-pathogen interface. Nature Review Microbiology. 9(8): 604-615.

Kiral F., Karagenc T., Pasa S., Yenisey C. & Seyrek K. 2005. Dogs with Hepatozoon canis respond to the oxidative stress by increased production of glutathione and nitric oxide. Veterinary Parasitology. 131(1): 15-21.

Lang T., Lecoeur H. & Prina E. 2009. Imaging Leishmania development in their host cells. Trends in Parasitology. 25(10): 465-473.

Limón-Pacheco J. & Gonsebatt M.E. 2009. The role of antioxidants and antioxidant-related enzymes in protective responses to environmentally induced oxidative stress. Mutation Research. 674(1): 137-147.

Lira R.A., Cavalcanti M.P., Nakazawa M., Ferreira A.G., Silva E.D., Abath F.G., Alves L.C., Souza W.V. & Gomes Y.M. 2006. Canine visceral leishmaniosis: a comparative analysis of the EIE-leishmaniose-visceral-canina-BioManguinhos and the IFI-leishmaniose-visceral-canina-Bio-Manguinhos kits. Veterinary Parasitology. 137(1): 11-16.

Lubos E., Loscalzo J. & Handy D.E. 2011. Glutathione peroxidase-1 in health and disease: from molecular mechanisms to therapeutic opportunities. Antioxidants & Redox Signaling. 15(7): 1958-1969.

Maia-Elkhoury A.N., Alves W.A., Sousa-Gomes M.L., Sena J.M. & Luna E.A. 2008. Leishmaniose visceral no Brasil: evolução e desafios. Cadernos de Saúde Pública. 24(12): 2941-2947.

Maia C. & Cardoso L. 2015. Spread of Leishmania infantum in Europe with dog travelling. Veterinary Parasitology. 213: 2-11.

Mol J.P.S., Soave S.A., Turchetti A.P., Pinheiro G.R.G., Pessanha A.T., Malta M.C.C., Tinoco H.P., Figueiredo L.A., Gontijo N.F., Paixão T.A., Fujiwara R.T. & Santos R.L. 2015. Transmissibility of Leishmania infantum from maned wolves (Chrysocyon brachyurus) and bush dogs (Speothos venaticus) to Lutzomyia longipalpis. Veterinary Parasitology. 212: 86-91.

Naito Y., Lee M.C., Kato Y., Nagai R. & Yonei Y. 2010. Oxidative stress markers. Anti-Aging Medicine. 7(5): 36-44.

Nathan C. & Cunningham-Bussel A. 2013. Beyond oxidative stress: an immunologist’s guide to reactive oxygen species. Nature Reviews: Immunology. 13(1): 349-361.

Nicolato C.R., Abreu R.T., Roatt B.M., Aguiar-Soares R.D., Reis L.E., Carvalho M.D., Carneiro C.M., Giunchetti R.C., Coura-Vital W. & Reis A.B. 2013. Clinical forms of canine visceral leishmaniasis in naturally Leishmania infantum-infected dogs and related myelogram and hemogram changes. PLoS One. 8(12): 1-9.

Panda D., Patra R.C., Nandi S. & Swarup D. 2009. Oxidative stress indices in gastroenteritis in dogs with canine parvoviral infection. Research in Veterinary Science. 86(3): 36-42.

Perego R., Proverbio D., Giorgi G.B. & Spada E. 2014. Prevalence of dermatological presentations of canine leishmaniasis in a nonendemic area: A retrospective study of 100 dogs. Veterinary Medicine International. 37(12): 1-5.

Silva F.S. 2007. Patologia e patogênese da leishmaniose visceral canina. Revista Trópica - Ciências Agrárias e Biológicas. 1(1): 20-31.

Singh S.K., Dimri U., Sharma M.C., Swarup D. & Sharma B. 2011. Determination of oxidative status and apoptosis in peripheral blood of dogs with sarcoptic mange. Veterinary Parasitology. 178(1): 330-338.

Zamocky M., Furtmüller P.G. & Obinger C. 2008. Evolution of catalases from bacteria to humans. Antioxidants & Redox Signaling. 10(9): 1527-1547.




DOI: https://doi.org/10.22456/1679-9216.80936

Copyright (c) 2018 Belarmino Eugênio Lopes-Neto, Glauco Jonas Lemos Santos, Adam Leal Lima, Maritza Cavalcante Barbosa, Talya Ellen Jesus dos Santos, Daniel Couto Uchoa, Ana Débora Nunes Pinheiro, Romélia Gonçalvez Pinheiro, Diana Célia Sousa Nunes-Pinheiro

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