Dhagma Renata Denis SOUZA, Andreia Lima Tome MELO, Livia Saab MURARO, Daniel Moura AGUIAR, Deijanira ALBUQUERQUE

Levamisole enhances global and diferential leukocyte numbers in peripheral blood of dogs with ehrlichiosis

Ehrlichiosis is a disease caused by Ehrlichia , a genus of obligatory intracellular bacteria that parasitize leukocytes and provokeseveral hematological alterations. In this work, we investigated the efect of levamisole associated with doxycycline on peripheralleukocytes in dogs PCR-positive for ehrlichial DNA. Twenty-one Ehrlichia -infected dogs were randomly distributed in 2 groups: acontrol group (n = 10) and a levamisole-treated group (n = 11). Blood samples were collected from each animal for hematological andDNA analysis both before initiating the experiment and 5 days later. All dogs were treated with doxycycline (10 mg/kg body weight),cimetidine (5 mg/kg of body weight), and fuid therapy for 5 consecutive days. In addition to receiving antibiotic therapy, the levamisole-treated dogs received a single subcutaneous injection (1 mg/kg body weight) of levamisole on the frst day of the experiment. Te resultsshow that levamisole associated with doxycycline signifcantly enhanced global leukocyte, lymphocyte, and monocyte numbers in theperipheral blood of dogs with ehrlichiosis. Tose data suggest that the combination of doxycycline with levamisole may improve canineehrlichiosis treatment through impairment of leukopenia, which is not controlled by antibiotic therapy alone.

PDF

___

1. Dumler JS, Barbet AF, Bekker CP, Dasch GA, Palmer GH, Ray SC, Rikihisa Y, Rurangirwa FR. Reorganization of genera in the families Rickettsiaceae and Anaplasmataceae in the order Rickettsiales: unifcation of some species of Ehrlichia with Anaplasma , Cowdria with Ehrlichia and Ehrlichia with Neorickettsia, descriptions of six new species combinations and designation of Ehrlichia equi and HE agent as subjective synonyms of Ehrlichia phagocytophila. Int J Syst Evol Microbiol 2001; 51: 21452165.
2. Maeda K, Markowitz N, Hawley RC, Ristic M, Cox D, McDade JE. Human infection with Ehrlichia canis, a leukocytic rickettsia. New Engl J Med 1987; 316: 853856.
3. Anderson BE, Dawson JED, Jones C, Wilson KH. Ehrlichia chafeensis, a new species associated with human ehrlichiosis. J Clin Microbiol 1991; 29: 28382842.
4. Perez M, Bodor M, Zhang C, Xiong Q, Rikihisa Y. Human infection with Ehrlichia canis accompanied by clinical signs in Venezuela. Ann NY Acad Sci 2006; 1078: 110117.
5. Cohn LA. Ehrlichiosis and related infections. Vet Clin North Am Small Anim Pract 2003; 33: 863884.
6. Dantas-Torres F. Canine vector-borne diseases. Brazil Parasit Vectors 2008; 1: 117.
7. Oliveira LS, Oliveira KA, Mourão LC, Pescatore AM, Almeida MR, Conceição LG, Galvão MA, Mafra C. First report of Ehrlichia ewingii detected by molecular investigation in dogs from Brazil. Clin Microbiol Infect 2009; 2: 5556.
8. Ewing SA. Canine ehrlichiosis. Adv Vet Sci Comp Med 1969; 13: 331353.
9. Huxsoll DL, Amyx HL, Hemelt IE, Hildebrandt PK, Nims RM, Gochenour WS Jr. Laboratory studies of tropical canine pancytopenia. Exp Parasitol 1972; 31: 5359.
10. Buhles WC Jr, Huxsoll DL, Ristic M. Tropical canine pancytopenia: clinical, hematologic, and serologic response of dogs to Ehrlichia canis infection, tetracycline therapy, and challenge inoculation. J Infect Dis 1974; 130: 357367.
11. Buhles WC Jr, Huxsoll DL, Hildebrandt PK. Tropical canine pancytopenia: role of aplastic anaemia in the pathogenesis of severe disease. J Comp Pathol 1975: 85: 511521.
12. Castro MB, Machado RZ, Aquino LPCT, Alessi AC, Costa MT. Experimental acute canine monocytic ehrlichiosis: clinicopathological and immunopathological fndings. Vet Parasitol 2004; 119: 7386.
13. Waner T, Harrus S, Bark H, Bogin E, Avidar Y, Keysary A. Characterization of the subclinical phase of canine ehrlichiosis in experimentally infected beagle dogs. Vet Parasitol 1997; 69: 307317.
14. Codner EC, Farris-Smith LL. Characterization of the subclinical phase of ehrlichiosis in dogs. J Am Vet Med Assoc 1986; 189: 4750.
15. Skotarczak B. Canine ehrlichiosis. Ann Agric Environ Med 2003; 10: 137141.
16. Mylonakis ME, Koutinas AF, Breitschwerdt EB, Hegarty BC, Billinis CD, Leontides LS, Kontos VS. Chronic canine ehrlichiosis (Ehrlichia canis): a retrospective study of 19 natural cases. J Am Anim Hosp Assoc 2004; 40: 174184.
17. Eddlestone SM, Diniz PP, Neer TM, Gaunt SD, Corstvet R, Cho D, Hosgood G, Hegarty B, Breitschwerdt EB. Doxycycline clearance of experimentally induced chronic Ehrlichia canis infection in dogs. J Vet Intern Med 2007; 21: 12371242.
18. McClure JC, Crothers ML, Schaefer JJ, Stanley PD, Needham GR, Ewing SA, Stich RW. Efcacy of a doxycycline treatment regimen initiated during three diferent phases of experimental ehrlichiosis. Antimicrob Agents Chemother 2010; 54: 50125020.
19. Neer TM, Harrus S. Canine monocytotropic ehrlichiosis and neorickettsiosis (E. canis, E. chafeensis, E. ruminantium , N. sennetsu , and N. risticii infections). In: Greene CE, editor. Infectious Diseases of the Dog and Cat. Philadelphia, PA, USA: Elsevier Saunders Company; 2006. pp. 203232.
20. Chen LY, Lin YL, Chiang BL. Levamisole enhances immune response by afecting the activation and maturation of human monocyte-derived dendritic cells. Clin Exp Immunol 2007; 151: 174181.
21. Zhang W, Du X, Zhao G, Jin H, Kang Y, Xiao C, Liu M, Wang B. Levamisole is a potential facilitator for the activation of T1 responses of the subunit HBV vaccination. Vaccine 2009; 27: 49384946.
22. Sangioni LA, Horta MC, Vianna MCB, Gennari SM, Soares RM, Galvão MAM, Schumaker TTS, Ferreira F, Vidotto O, Labruna MB. Rickettsial infection in animals and Brazilian Spotted Fever endemicity. Emerg Infec Dis 2005; 11: 265270.
23. Labruna MB, McBride JW, Camargo LM, Aguiar DM, Yabsley MJ, Davidson WR, Stromdahl EY, Williamson PC, Stich RW, Long SW et al. A preliminary investigation of Ehrlichia species in ticks, humans, dogs, and capybaras from Brazil. Vet Parasitol 2007; 143: 189195.
24. Sousa MCBB. Infuência da Dose e Esquema de Administração de C. parvum Inativado, Cepa DAUFEPE nº 187, Sobre o Eritograma, Leucograma e Perfl Protéico de Camundongos (Mus musculos). MSc, Universidade Federal Rural de Pernambuco, Recife, PE, Brazil, 1992 (in Portuguese).
25. Tienpont D, Vanparijs OF, Raeymaekers AH, Vandenberk J, Demoen JA, Allewijn FT, Marsboom RP, Niemegeers CJ, Schellekens KH, Janssen PA. Tetramisole (R 8299), a new, potent broad spectrum anthelmintic. Nature 1966; 209: 1084 1086.
26. Mutch RS, Hutson PR. Levamisole in the adjuvant treatment of colon cancer. Clin Pharm 1991; 10: 95109.
27. Katoch K. Immunotherapy of leprosy. Indian J Lepr 1996; 68: 349361.
28. Kocabas CN, Sekerel BE, Firat PA, Okur H, Adahoglu G. Levamisole: might it be used in treatment and prevention of atopic diseases? J Asthma 2004; 41: 547551.
29. Kobayashi M, Fitz L, Ryan M, Hewick RM, Clark SC, Chan S, Loudon R, Sherman F, Perussia B, Trinchieri G. Identifcation and purifcation of natural killer cell stimulatory factor (NKSF), a cytokine with multiple biologic efects on human lymphocytes. J Exp Med 1989; 170: 827845.
30. Rodrigues OG, Sousa MRQ, Duarte MHS, Castro RS. Avaliação da Infuência de Levamisole sobre Padrões de Proteínas e Leucograma em caprinos (Capra hircus) nativos da Região Semi-Arida Nordestina. Agropecuária Científca no Semi-árido 2005; 1: 5058 (in Portuguese).
31. Xavier MS, Almosny NRP, Nascimento MD, Silva GVO, Botelho GG. Avaliação da coagulação plasmática e plaquetometria em cães não infectados e infectados experimentalmente com Ehrlichia spp. Arq Bras Med Vet Zootec 2009; 61: 10491053 (in Portuguese).