diagnosco terapeuco gesonaleinassenzadicrisileishmanioca ... · introduction: canine visceral...

Relazione Prof. G. Lubas 12/11/17

FORMEV, Bari 12 Nov. 2017 1

Gruppo di Studio sulla Leishmaniosi Canina (GSLC) ObieEvi e Finalità

•  Razionale e omogeneo approccio al paziente leishmanioMco

•  Fondazione nella prima riunione a Roma (26 Nov 2005)

•  Rivolta ad oEmizzare, nella leishmaniosi del cane, i seguenM aspeE: – DiagnosMco –  TerapeuMco – GesMonale in assenza di crisi leishmanioMca –  Prevenzione


Composizione del GSLC •  Chairman, Prof. George Lubas (Dipl ECVIM-‐CA, Universita di Pisa) •  Dermatologa, DoY.ssa Alessandra FondaM (Dipl ECVD, libero

professionista, Roma) •  Patologo clinico, Prof. Saverio Paltrinieri (Dipl ECVCP, Univ. Milano) •  Nefrologo DoY. Andrea Zatelli (Medical Consultancy Services, Malta) •  Internista, DoY. Xavier Roura (Dipl ECVIM-‐CA, Univ. Barcellona, E) •  Nefrologo, DoY. Eric Zini (Dipl ECVIM-‐CA, Univ. di Padova) •  Epidemiologo, Dr. Michele Maroli (ex IsMt. Super. Sanità, Roma) •  Parassitologo, Dr. Luigi Gradoni (IsMtuto Superiore Sanità, Roma)





Sito web: www.gruppoleishmania.org

UMlità degli esami ematobiochimici nel cane affeYo da leishmaniosi

George Lubas Prof. Ord. Clinica Medica Veterinaria

Dipl. ECVIM-‐CA




Sommario

Descrivere in corso di leishmaniosi le anormalità clinico-‐patologiche su:

•  Emogramma – Midollo osseo

•  Profilo biochimico – sieroeleYroforesi

•  Esame delle urine •  Profilo coagulaMvo


Emogramma completo

•  Eritrone – RBC, Hgb, Hct, MCV, MCHC, RDW, morfologia

•  Leucone – WBC; VA: Neu, Lin, Mon, Eos, Bas; morfologia

•  Trombone – PLT, MPV, morfologia


Paltrinieri et al, 2016



Leucone

•  Leucogramma infiammatorio (Kiral et al, 2004; Nicolato et al, 2013) – éNeu + Mon – êLin; – êéEos

•  Possibilità di rinvenire amasMgoM? (De Gopegui & Espada, 1998; Foglia Manzillo et al, 2005; Giudice & PassanMno, 2011)


AMASTIGOTES IN CIRCULATING LEUKOCYTES OF DOGS WITH LEISHMANIOSIS 207

Acta Veterinaria Hungarica 59, 2011

[RR: 42.5–77.3%; 3.9–8.0 × 109/L]), with right shift of the Arneth index (the nu-

cleus showed more filiform lobes, indicating hypersegmentation); several round to

oval basophilic intracytoplasmic inclusions (1–5 per cell), identified as Leishmania

sp. amastigotes, occurring in 12% of mature circulating neutrophils (Fig. 1) and

numerous free parasites. Anaplasma (Ehrlichia) platys-like deep blue inclusions

within approximately 2% of the platelets were also observed. Lymph node biopsy

showed several Leishmania amastigotes.

Table 2

Major clinical and laboratory findings in 4 dogs

Findings Case 1 Case 2 Case 3 Case 4

Depression x x x

Anorexia/dysorexia x x x

Weigh loss x x x x

Dehydration x x

Pale mucous membranes x x

Jaundice x

Onychogryphosis x

Dermatitis x

Lymphadenopathy x x x x

Hepatosplenomegaly x x x x

Weakness x x

Paresis x

Hyperthermia x x x

Diarrhoea x

Anaemia x x x x

Leukocytosis x

Thrombocytopenia x x x x

Renal failure x x

Hepatic failure x x x

Dysproteinaemia x x x

Survival (days) 6 19 Alive 1

Figs 1A and 1B. Peripheral blood smear (May-Grünwald–Giemsa stain) from a dog (Case 1):

Leishmania amastigotes (arrows) within neutrophils

A B

210 GIUDICE and PASSANTINO

Acta Veterinaria Hungarica 59, 2011

The dog was given allopurinol (25 mg/kg/day per os), doxycycline (10 mg/ kg/day per os for 21 days) and hepatoprotectors. Two months later, the haema-tologic picture worsened (RBC 2.35 × 1012/L; Hb 4 g/dL; PCV 13.1%; PLT 89 × 109/L) and a blood transfusion was carried out. Lymph node fine needle aspirate revealed numerous intra-macrophagic amastigotes, despite allopurinol treatment. N-methylglucamine antimoniate (100 mg/kg/day sc. for 60 days) was associated to allopurinol, with a rapid improvement and regression of symptomatology.

Case 4

The blood smear of a 3-year-old male mongrel, with a history of 1-week depression, dysorexia and fever, was referred to our laboratory for haematologi-cal consultation. Clinical features observed by the external veterinarian consisted of weight loss, pale mucous membranes, lymphadenopathy, hepatosplenomegaly, renal failure, anaemia and thrombocytopenia. The dog died the following day. Mi-croscopic examination of the peripheral blood smear showed a single circulating macrophage with a high-number of intra-cytoplasmic Leishmania amastigotes (Fig. 4).

Fig. 4. Peripheral blood smear from a dog (Case 4): macrophage with engulfed amastigotes

The clinical signs of leishmaniosis vary widely as a result of the numerous

factors involved in the disease, the host’s immunological profile being one of the most important. Although the amastigotes can be detected within mononucleated phagocytes in several organs, their record on peripheral blood smears is rare and it is generally associated with severe illness and co-infections (Ruiz de Gopegui and Espada, 1998; Foglia Manzillo et al., 2005).

Eritrone

•  Anemia lieve-‐moderata normociMca-‐normocromica (ACI) scarsamente rigeneraMva

•  Patogenesi: –  Ipoproduzione (diseritropoiesi) (Kiral et al, 2004; De Tommasi et al, 2014)

– RidoYa EPO – EmoliMca (immunomediata secondaria) (Ciaramella et al, 1997)




Trombone •  Trombocitopenia lieve-‐moderata •  Patogenesi:

–  Immunomediata secondaria periferica (Cortese et al, 2009 & 2011; Terrazzano et al, 2006)

–  Consumo in associazione a êAT o DIC (Honse et al, 2013) –  Ipoproduzione (De Tommasi et al, 2014)

–  Sequestro ? –  Diminuita funzione PLT (trombocitopaMa?)

(Abid et al, 2015, Ciaramella et al, 2004))

AYenzione alle co-‐infezioni! Ehrlichia, Anaplasma


Midollo osseo (citologia)

•  Esame diagnosMco qualitaMvo/quanMtaMvo (De Abreu et al, 2011; Gavazza et al, 2002; Momo et al, 2014; Paparcone et al, 2013)

–  Ipoplasia eritroide (pool proliferaMvo e maturaMvo)

–  Iperplasia mieloide (éM:E) –  Iperplasia megacariocitaria

•  Infiltrato macrofagico (+/-‐ Leishmania) e plasmacellulare

•  Segni di displasia eritroide/mieloide/megacariocitaria (De Tommasi et, 2014; Foglia Manzillo et al, 2006)


(De Abreu et al, 2001; Foglia Manzillo et al, 2006; Momo et al, 2014; Nicolato et al, 2013; Sardomichelakis et al, 2005)



Profilo biochimico

•  Valutazione reazione infiammatoria/immune •  Valutazione della funzione renale

– Esame delle urine •  Altre valutazioni

– Epatobiliare – PancreaMca – Muscolo-‐scheletrica – Miocardica – Endocrina


Profilo Biochimico Infiammazione/Immune

Iperpro5demia •  APP posi5ve:

–  Prot. C reaEva, CRP –  Aptoglobulina, Hpt –  Ceruplasmina, Cp –  Amiloide sierica, SAA –  FerriMna

Ipoalbuminemia (êA/G) •  APP nega5ve:

–  Albumina –  Transferrina totale, TIBC –  Ferro, Fe –  Paraxonase 1, PON-‐1 –  HDL


Sieroele>roforesi: •  Ipoalbuminemia •  é Alfa-‐2 globuline •  é Beta globuline •  é Gamma globuline

Foglia Manzillo et al, 2013; KouMnas et al, 1999; Ribeiro et al, 2013;

Ibba et al, 2015; MarMnez-‐Subiela et al, 2002, 2003 2011, 2014; Rossi et al, 2013 ; Sasanelli et al, 2007; Silvestrini et al, 2014)



Profilo biochimico Rene

Ema5ci •  Urea •  CreaMnina •  DimeMlarginina simmetrica,

SDMA

Urinari •  Peso specifico •  Sedimento aEvo •  Presenza cilindri •  Proteinuria •  PU/CU •  EleYroforesi urinaria, SDS-‐

PAGE/AGE •  Enzimuria (GGT)


(Aresu et al, 2013; Braga et al, 2015; Ibba et al, 2016; KouMnas & KouMnas, 2014; Zatelli et al, 2003)

Profilo biochimico Altre valutazioni

•  Epatobiliare –  Enzimi colestaMci, citoliMci, prove funzione epaMca

(Rallis et al, 2005; Tonin et al, 2016)

•  PancreaMca – Amilasi, lipasi, PLI

•  Muscolo scheletrica –  CK, LDH, AST (Vamvakidis et al, 2000)

•  Miocardica –  CK, troponina (Silvestrini et, 2012)

•  Endocrina -‐ IpoadrenocorMcismo (Momo et al, 2014)




Profilo coagulaMvo

•  Spesso presente ipercoagulabilità da: – êAT è trombosi è DIC –  Iperviscosità

•  Raramente alterato con ipocoagulabilità per: –  Interferenza pre-‐analiMca per discrasia A/G – Presenza DIC (Honse et al, 2013)



24

Revista da Sociedade Brasileira de Medicina Tropical 45(1):24-29, jan-fev, 2012

INTRODUCTION

Article/Artigo

1. Programa de Pós-Graduação em Ciências Veterinárias, Faculdade de Veterinária, Universidade Estadual do Ceará, Fortaleza, CE. 2. Núcleo Regional de Ofiologia, Universidade Federal do Ceará, Fortaleza, CE. 3. Laboratório de Fisiofarmacologia Cardio-Renal, Instituto Superior de Ciências Biomédicas, Universidade Estadual do Ceará, Fortaleza, CE. 4. Centro de Controle de Zoonoses de Fortaleza, Fortaleza, CE.Address to: Dr. José Claudio Carneiro de Freitas. Programa de Pós-Graduação em Ciências Veterinárias/FAVET/UECE. Av. Paranjana 1700, Campus do Itaperi, Serrinha, 60740-000 Fortaleza, CE, Brasil.Phone: 55 85 31019840e-mail: [email protected] in 21/01/2011Accepted in 29/09/2011

Clinical and laboratory alterations in dogs naturally infected by Leishmania chagasi

Alterações clínicas e laboratoriais em cães naturalmente infectados por Leishmania chagasi.

José Cláudio Carneiro de Freitas1, Diana Célia Sousa Nunes-Pinheiro1, Belarmino Eugênio Lopes Neto1, Glauco Jonas Lemos Santos1, Cyntia Rafaelle Amaral de Abreu1, Roberta Rocha Braga2, Rafael de Morais Campos3 and Ligene Fernandes de Oliveira4

ABSTRACTIntroduction: Canine visceral leishmaniasis (CVL) is a zoonotic disease with different clinical manifestations. Parasitism often occurs in bone marrow, but changes have been observed in peripheral blood and serum biochemical parameters. The aim of this study was to evaluate the hematological and biochemical parameters in dogs naturally infected by Leishmania chagasi. Methods: Eighty-five adult dogs of both sexes and various weights and ages from the Zoonosis Control Center of Fortaleza (CCZ) were used, selected by immunofluorescence assay (IFA) and considered positive with IFA titers greater than 1:40 and by visualizing amastigotes of Leishmania chagasi in smears obtained by bone marrow aspiration. The dogs (n = 85) were grouped according to clinical signs: negative (CN = 7), subclinical (CS = 10), and clinical (CC = 68). Blood samples were collected for determination of hematological and biochemical serum values. The experimental protocol was approved by the CEUA/UECE. Results: The most frequent clinical signs were cachexia (77.9%), keratitis (61.8%), and lymphadenopathy (55.9%), and 86.8% of the animals showed more than one clinical sign characteristic of CVL. In CC were observed reductions in red blood cells (63%), hematocrit (72%), and hemoglobin (62%), as well as leukocytosis (33%), neutropenia (28%), thrombocytopenia (50%), uremia (45%), hyperproteinemia (53%, p<0.05), hypergammaglobulinemia (62%, p<0.01), and hypoalbuminemia (58%). Conclusions: Animals with the clinical form of the disease demonstrate hematological and biochemical changes consistent with anemia, uremia, hyperproteinemia, and hyperglobulinemia, which present themselves as strong clinical markers of visceral leishmaniasis associated with the signs previously reported.Keywords: Dogs. Canine visceral leishmanisis. Biomarkers. Anemia. Uremia. Hyperglobulinemia.

RESUMOIntrodução: A leishmaniose visceral canina (LVC) é uma zoonose com diferentes manifestações clínicas. O parasitismo ocorre frequentemente na medula óssea e têm sido relatadas alterações hematológicas e bioquímicas. Objetivou-se avaliar os parâmetros clínicos, hematológicos e bioquímicos de cães naturalmente infectados por Leishmania chagasi. Métodos: Utilizaram-se 85 cães adultos, ambos os sexos, peso e idade variados, oriundos do Centro de Controle de Zoonoses de Fortaleza, selecionados pela reação de imunofluorescência indireta (RIFI), sendo considerados positivos os animais com títulos de RIFI ≥ 1:40 e pelo exame parasitológico das formas amastigotas de Leishmania chagasi em esfregaços de medula óssea. Os cães foram agrupados conforme os sinais clínicos associados à doença: negativos (CN=7); subclínicos (CS=10) e clínicos (CC=68). Amostras de sangue foram coletadas para determinação dos parâmetros hematológicos e bioquímicos séricos. O protocolo experimental foi aprovado pelo CEUA/UECE, protocolo n° 08622833-1. Resultados: Os sinais clínicos mais frequentes foram caquexia (77,9%), ceratoconjuntivite (61,8%) e linfadenopatia (55,9%), sendo que 86, 8% dos animais apresentaram mais de um sinal clínico característico de LVC. Em CC foram observadas reduções nas hemácias (63%), hematócrito (72%) e hemoglobina (62%), leucocitose (33%), neutropenia (28%), trombocitopenia (50%), uremia (45%), hiperproteinemia (53%, p<0,05), hiperglobulinemia (62%, p<0,01) e hipoalbuminemia (58%). Conclusões: Concluiu-se que os animais com a forma clínica da doença apresentam alterações condizentes com anemia, uremia, hiperproteinemia e hiperglobulinemia, as quais se apresentam como marcadores clínicos da leishmaniose visceral, associados aos sinais previamente relatados. Palavras-chaves: Cães. Leishmaniose visceral canina. Biomarcadores. Anemia. Uremia. Hiperglobulinemia.

Visceral leishmaniasis is a zoonosis that affects humans when they come into contact with the transmission cycle of the parasite1. It is one of the most relevant emerging diseases worldwide, and Brazil is among the countries of Latin America that present the greatest number of human cases, about 90% of annual cases2.

Although humans can also act as reservoirs of the agent and play a role in the transmission cycle, the dog is considered one of the most important links in the epidemiological chain of leishmaniasis3. Canine visceral leishmaniasis (CVL) is transmitted through the bite of insects known as sandflies, mainly the species Lutzomyia longipalpis and L. cruzi, which convey the infective promastigotes. The main agent of visceral leishmaniasis in Brazil is Leishmania (Leishmania) chagasi1,4.

The pathogenesis of CVL involves several factors, and a decisive factor in the disease progression is associated with the immune response that the animal develops against the parasite5-7. In this case, the antibodies, rather than having a protective function, become highly harmful, participating in inflammatory processes and being responsible for most of the clinical signs associated with CVL6,8,9.

The infection may present itself in clinical form (clinical dogs), in which dogs show clinical signs and/or typical clinical and laboratory changes with confirmation of Leishmania chagasi, or in subclinical form (subclinical dogs), in which dogs show no clinical and laboratory changes, but the presence of Leishmania chagasi is confirmed by routine diagnostic tests10.

The hematological and serum biochemical parameters, although limited in the diagnosis of CVL, are very useful in evaluating the clinical status of the animal and the extent of lesions and might give indications on the animal prognosis11,12. However, there is little information on these parameters and on biomarkers of leishmaniasis.

Considering the relevance of the disease and the scarcity of information about the clinical parameters

24

Revista da Sociedade Brasileira de Medicina Tropical 45(1):24-29, jan-fev, 2012

INTRODUCTION

Article/Artigo

1. Programa de Pós-Graduação em Ciências Veterinárias, Faculdade de Veterinária, Universidade Estadual do Ceará, Fortaleza, CE. 2. Núcleo Regional de Ofiologia, Universidade Federal do Ceará, Fortaleza, CE. 3. Laboratório de Fisiofarmacologia Cardio-Renal, Instituto Superior de Ciências Biomédicas, Universidade Estadual do Ceará, Fortaleza, CE. 4. Centro de Controle de Zoonoses de Fortaleza, Fortaleza, CE.Address to: Dr. José Claudio Carneiro de Freitas. Programa de Pós-Graduação em Ciências Veterinárias/FAVET/UECE. Av. Paranjana 1700, Campus do Itaperi, Serrinha, 60740-000 Fortaleza, CE, Brasil.Phone: 55 85 31019840e-mail: [email protected] in 21/01/2011Accepted in 29/09/2011

Clinical and laboratory alterations in dogs naturally infected by Leishmania chagasi

Alterações clínicas e laboratoriais em cães naturalmente infectados por Leishmania chagasi.

José Cláudio Carneiro de Freitas1, Diana Célia Sousa Nunes-Pinheiro1, Belarmino Eugênio Lopes Neto1, Glauco Jonas Lemos Santos1, Cyntia Rafaelle Amaral de Abreu1, Roberta Rocha Braga2, Rafael de Morais Campos3 and Ligene Fernandes de Oliveira4

ABSTRACTIntroduction: Canine visceral leishmaniasis (CVL) is a zoonotic disease with different clinical manifestations. Parasitism often occurs in bone marrow, but changes have been observed in peripheral blood and serum biochemical parameters. The aim of this study was to evaluate the hematological and biochemical parameters in dogs naturally infected by Leishmania chagasi. Methods: Eighty-five adult dogs of both sexes and various weights and ages from the Zoonosis Control Center of Fortaleza (CCZ) were used, selected by immunofluorescence assay (IFA) and considered positive with IFA titers greater than 1:40 and by visualizing amastigotes of Leishmania chagasi in smears obtained by bone marrow aspiration. The dogs (n = 85) were grouped according to clinical signs: negative (CN = 7), subclinical (CS = 10), and clinical (CC = 68). Blood samples were collected for determination of hematological and biochemical serum values. The experimental protocol was approved by the CEUA/UECE. Results: The most frequent clinical signs were cachexia (77.9%), keratitis (61.8%), and lymphadenopathy (55.9%), and 86.8% of the animals showed more than one clinical sign characteristic of CVL. In CC were observed reductions in red blood cells (63%), hematocrit (72%), and hemoglobin (62%), as well as leukocytosis (33%), neutropenia (28%), thrombocytopenia (50%), uremia (45%), hyperproteinemia (53%, p<0.05), hypergammaglobulinemia (62%, p<0.01), and hypoalbuminemia (58%). Conclusions: Animals with the clinical form of the disease demonstrate hematological and biochemical changes consistent with anemia, uremia, hyperproteinemia, and hyperglobulinemia, which present themselves as strong clinical markers of visceral leishmaniasis associated with the signs previously reported.Keywords: Dogs. Canine visceral leishmanisis. Biomarkers. Anemia. Uremia. Hyperglobulinemia.

RESUMOIntrodução: A leishmaniose visceral canina (LVC) é uma zoonose com diferentes manifestações clínicas. O parasitismo ocorre frequentemente na medula óssea e têm sido relatadas alterações hematológicas e bioquímicas. Objetivou-se avaliar os parâmetros clínicos, hematológicos e bioquímicos de cães naturalmente infectados por Leishmania chagasi. Métodos: Utilizaram-se 85 cães adultos, ambos os sexos, peso e idade variados, oriundos do Centro de Controle de Zoonoses de Fortaleza, selecionados pela reação de imunofluorescência indireta (RIFI), sendo considerados positivos os animais com títulos de RIFI ≥ 1:40 e pelo exame parasitológico das formas amastigotas de Leishmania chagasi em esfregaços de medula óssea. Os cães foram agrupados conforme os sinais clínicos associados à doença: negativos (CN=7); subclínicos (CS=10) e clínicos (CC=68). Amostras de sangue foram coletadas para determinação dos parâmetros hematológicos e bioquímicos séricos. O protocolo experimental foi aprovado pelo CEUA/UECE, protocolo n° 08622833-1. Resultados: Os sinais clínicos mais frequentes foram caquexia (77,9%), ceratoconjuntivite (61,8%) e linfadenopatia (55,9%), sendo que 86, 8% dos animais apresentaram mais de um sinal clínico característico de LVC. Em CC foram observadas reduções nas hemácias (63%), hematócrito (72%) e hemoglobina (62%), leucocitose (33%), neutropenia (28%), trombocitopenia (50%), uremia (45%), hiperproteinemia (53%, p<0,05), hiperglobulinemia (62%, p<0,01) e hipoalbuminemia (58%). Conclusões: Concluiu-se que os animais com a forma clínica da doença apresentam alterações condizentes com anemia, uremia, hiperproteinemia e hiperglobulinemia, as quais se apresentam como marcadores clínicos da leishmaniose visceral, associados aos sinais previamente relatados. Palavras-chaves: Cães. Leishmaniose visceral canina. Biomarcadores. Anemia. Uremia. Hiperglobulinemia.

Visceral leishmaniasis is a zoonosis that affects humans when they come into contact with the transmission cycle of the parasite1. It is one of the most relevant emerging diseases worldwide, and Brazil is among the countries of Latin America that present the greatest number of human cases, about 90% of annual cases2.

Although humans can also act as reservoirs of the agent and play a role in the transmission cycle, the dog is considered one of the most important links in the epidemiological chain of leishmaniasis3. Canine visceral leishmaniasis (CVL) is transmitted through the bite of insects known as sandflies, mainly the species Lutzomyia longipalpis and L. cruzi, which convey the infective promastigotes. The main agent of visceral leishmaniasis in Brazil is Leishmania (Leishmania) chagasi1,4.

The pathogenesis of CVL involves several factors, and a decisive factor in the disease progression is associated with the immune response that the animal develops against the parasite5-7. In this case, the antibodies, rather than having a protective function, become highly harmful, participating in inflammatory processes and being responsible for most of the clinical signs associated with CVL6,8,9.

The infection may present itself in clinical form (clinical dogs), in which dogs show clinical signs and/or typical clinical and laboratory changes with confirmation of Leishmania chagasi, or in subclinical form (subclinical dogs), in which dogs show no clinical and laboratory changes, but the presence of Leishmania chagasi is confirmed by routine diagnostic tests10.

The hematological and serum biochemical parameters, although limited in the diagnosis of CVL, are very useful in evaluating the clinical status of the animal and the extent of lesions and might give indications on the animal prognosis11,12. However, there is little information on these parameters and on biomarkers of leishmaniasis.

Considering the relevance of the disease and the scarcity of information about the clinical parameters




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Relationship between clinical and pathological signs and severity of canine leishmaniasis

Relação entre sinais clínico-patológicos e gravidade da leishmaniose visceral canina

Raul Rio Ribeiro1*; Sydnei Magno da Silva2; Gustavo de Oliveira Fulgêncio3; Marilene Suzan Marques Michalick4; Frédéric Jean Georges Frézard5

1Centro de Ciências Agrárias, Ambientais e Biológicas, Universidade Federal do Recôncavo da Bahia – UFRB, Cruz das Almas, BA, Brasil2Departamento de Imunologia, Microbiologia e Parasitologia, Instituto de Ciências Biomédicas, Universidade Federal de Uberlândia – UFU, Uberlândia, MG, Brasil

3Departamento de Clínica e Cirurgia Veterinárias, Escola de Veterinária, Universidade Federal de Minas Gerais – UFMG, Belo Horizonte, MG, Brasil

4Laboratório de Sorologia, Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais – UFMG, Belo Horizonte, MG, Brasil

5Laboratório de Biofísica e Sistemas Nanoestruturados, Departamento de Fisiologia e Biofísica, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais – UFMG, Belo Horizonte, MG, Brasil

Received March 10, 2013 Accepted May 28, 2013

Abstract

Canine visceral leishmaniasis (CVL) is a zoonotic disease that presents variable clinical and laboratory aspects. The aims of this study were to identify the main biochemical/hematological status of dogs naturally infected with Leishmania (Leishmania) infantum and to associate theses parameters with clinical forms of CVL. Blood samples were analyzed from 51 dogs, 15 uninfected (control group) and 36 infected, which were classified clinically in three groups: asymptomatic (n=12), oligosymptomatic (n=12) and symptomatic (n=12). All the infected dogs showed lower albumin/globulin ratios (A-G ratio) than the limit of reference. The mean values of total protein, urea, α-globulin 2, globulin and A-G ratio of infected dogs were outside the reference interval and differed significantly from those of the controls. Anemia was detected only in groups that showed clinical signs of the disease, and a statistical analysis indicated a significantly higher frequency of lower eritrogram in these groups than in the asymptomatic group. In addition, a significant association was observed between anemia and the presence of the symptoms, with dogs displaying higher erythrogram values showing better clinical conditions. These results provide additional evidence that the clinical forms of CVL may reflect on the erythrogram status.

Keywords: Canine visceral leishmaniasis, clinical forms, laboratory findings, anaemia, relationship.

Resumo

A leishmaniose visceral canina (LVC) é uma zoonose com aspectos clínicos e laboratoriais variáveis. O objetivo deste trabalho foi identificar os principais achados hematológicos e bioquímicos em cães naturalmente infectados com Leishmania (Leishmania) infantum e associar esses parâmetros com as formas clínicas da LVC. Foram analizadas amostras sanguíneas provenientes de 51 cães, sendo 15 cães não infectados (grupo controle) e 36 infectados, os quais foram classificados clinicamente em três grupos: assintomáticos (n=12), oligossintomáticos (n=12) e sintomáticos (n=12). Todos os cães infectados apresentaram valores na relação albumina/globulina (A/G) abaixo do limite inferior de referência. Os valores médios de proteína total, uréia, α-2 globulina, globulina e A/G dos grupos de cães infectados permaneceram fora dos intervalos de referências e significativamente diferente quando comparados aos do grupo controle. Anemia foi registrada somente nos grupos de animais que manifestavam sinais clínicos da enfermidade, sendo que nas análises estatísticas constatou-se frequencia significativamente maior de alterações no eritrograma quando comparados ao grupo assintomático. Associação significativa foi observada entre anemia e a presença de sinais clínicos, onde os cães com os maiores valores de eritrograma apresentavam a melhor condição clínica. Os resultados fornecem evidência adicional que as formas clínicas da LVC podem refletir no eritrograma.

Palavras-chave: Leishmaniose visceral canina, formas clínicas, achados laboratoriais, anemia, relação.

*Corresponding author: Raul Rio Ribeiro Centro de Ciências Agrárias, Ambientais e Biológicas, Universidade Federal do Recôncavo da Bahia – UFRB, CEP 44380-000, Cruz das Almas, BA, Brazil e-mail: [email protected]

Full Article

ISSN 0103-846X (impresso) / ISSN 1984-2961 (eletrônico)Rev. Bras. Parasitol. Vet., Jaboticabal, v. 22, n. 3, p. 373-378, jul.-set. 2013

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Relationship between clinical and pathological signs and severity of canine leishmaniasis

Relação entre sinais clínico-patológicos e gravidade da leishmaniose visceral canina

Raul Rio Ribeiro1*; Sydnei Magno da Silva2; Gustavo de Oliveira Fulgêncio3; Marilene Suzan Marques Michalick4; Frédéric Jean Georges Frézard5

1Centro de Ciências Agrárias, Ambientais e Biológicas, Universidade Federal do Recôncavo da Bahia – UFRB, Cruz das Almas, BA, Brasil2Departamento de Imunologia, Microbiologia e Parasitologia, Instituto de Ciências Biomédicas, Universidade Federal de Uberlândia – UFU, Uberlândia, MG, Brasil

3Departamento de Clínica e Cirurgia Veterinárias, Escola de Veterinária, Universidade Federal de Minas Gerais – UFMG, Belo Horizonte, MG, Brasil

4Laboratório de Sorologia, Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais – UFMG, Belo Horizonte, MG, Brasil

5Laboratório de Biofísica e Sistemas Nanoestruturados, Departamento de Fisiologia e Biofísica, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais – UFMG, Belo Horizonte, MG, Brasil

Received March 10, 2013 Accepted May 28, 2013

Abstract

Canine visceral leishmaniasis (CVL) is a zoonotic disease that presents variable clinical and laboratory aspects. The aims of this study were to identify the main biochemical/hematological status of dogs naturally infected with Leishmania (Leishmania) infantum and to associate theses parameters with clinical forms of CVL. Blood samples were analyzed from 51 dogs, 15 uninfected (control group) and 36 infected, which were classified clinically in three groups: asymptomatic (n=12), oligosymptomatic (n=12) and symptomatic (n=12). All the infected dogs showed lower albumin/globulin ratios (A-G ratio) than the limit of reference. The mean values of total protein, urea, α-globulin 2, globulin and A-G ratio of infected dogs were outside the reference interval and differed significantly from those of the controls. Anemia was detected only in groups that showed clinical signs of the disease, and a statistical analysis indicated a significantly higher frequency of lower eritrogram in these groups than in the asymptomatic group. In addition, a significant association was observed between anemia and the presence of the symptoms, with dogs displaying higher erythrogram values showing better clinical conditions. These results provide additional evidence that the clinical forms of CVL may reflect on the erythrogram status.

Keywords: Canine visceral leishmaniasis, clinical forms, laboratory findings, anaemia, relationship.

Resumo

A leishmaniose visceral canina (LVC) é uma zoonose com aspectos clínicos e laboratoriais variáveis. O objetivo deste trabalho foi identificar os principais achados hematológicos e bioquímicos em cães naturalmente infectados com Leishmania (Leishmania) infantum e associar esses parâmetros com as formas clínicas da LVC. Foram analizadas amostras sanguíneas provenientes de 51 cães, sendo 15 cães não infectados (grupo controle) e 36 infectados, os quais foram classificados clinicamente em três grupos: assintomáticos (n=12), oligossintomáticos (n=12) e sintomáticos (n=12). Todos os cães infectados apresentaram valores na relação albumina/globulina (A/G) abaixo do limite inferior de referência. Os valores médios de proteína total, uréia, α-2 globulina, globulina e A/G dos grupos de cães infectados permaneceram fora dos intervalos de referências e significativamente diferente quando comparados aos do grupo controle. Anemia foi registrada somente nos grupos de animais que manifestavam sinais clínicos da enfermidade, sendo que nas análises estatísticas constatou-se frequencia significativamente maior de alterações no eritrograma quando comparados ao grupo assintomático. Associação significativa foi observada entre anemia e a presença de sinais clínicos, onde os cães com os maiores valores de eritrograma apresentavam a melhor condição clínica. Os resultados fornecem evidência adicional que as formas clínicas da LVC podem refletir no eritrograma.

Palavras-chave: Leishmaniose visceral canina, formas clínicas, achados laboratoriais, anemia, relação.

*Corresponding author: Raul Rio Ribeiro Centro de Ciências Agrárias, Ambientais e Biológicas, Universidade Federal do Recôncavo da Bahia – UFRB, CEP 44380-000, Cruz das Almas, BA, Brazil e-mail: [email protected]

Full Article

ISSN 0103-846X (impresso) / ISSN 1984-2961 (eletrônico)Rev. Bras. Parasitol. Vet., Jaboticabal, v. 22, n. 3, p. 373-378, jul.-set. 2013


R.B.P. Torrecilha et al. / Preventive Veterinary Medicine 132 (2016) 83–87 85

Fig. 1. Correlogram of Spearman’s rank correlation coefficients between Leishmania spp. density (horizontal axis) in different tissues and clinical alterations, blood cell count,biochemical and oxidative stress markers (vertical axis). Warm colors (towards red) represent positive correlations (r > 0), whereas cool colors (towards blue) represent nega-tive correlations (r < 0). Only markers presenting at least one suggestive (0.05 ≤ P < 0.10) or significant (P < 0.05) correlation in a randomization test with 10,000 permutationsare shown. For a complete summary of correlations see the Supplementary information (Supplementary Table 3 and 4). (For interpretation of the references to colour in thisfigure legend, the reader is referred to the web version of this article.)

and skin samples, respectively. However, all dogs presented para-site infection in at least one tissue, confirming that all dogs wereinfected. Parasite load across tissues were moderately to highlycorrelated, with an average correlation of r = 0.400 (Supplemen-tary information). An exception was observed for blood, which waspoorly correlated with all tissues (average correlation of r = 0.069).

In order to have a representation of all tissues simultaneously,we created a new compressed variable, namely CPLD, based on aprincipal component analysis of the PL matrix. For that matter, werequired complete observations of PL across tissues, which reducedthe data from 37 to 31 samples. The first leading principal compo-nent explained 49.1% of the total variance contained in the originalmatrix, and therefore was adopted as the CPLD. The relative con-tributions of each tissue to the CPLD were 22.7% for skin, 28.5% forright conjunctiva, 29.6% for left conjunctiva, 12.3% for buffy coatand 6.9% for blood.

3.4. Correlations for peripheral parasite load

As the permutation test is conservative, we decided to discusssuggestive (0.05 ≤ P < 0.10) and significant (P < 0.05) correlations.All clinical alterations were positively correlated (P < 0.05) withparasite load in two or more tissues (Fig. 1). Ophthalmic alterationswere only associated with PL in the conjunctivas. Parasite load inthe blood did not present associations with clinical alterations. TheCPLD variable successfully captured the correlatons of the individ-ual tissues, and only failed to predict ophthalmic alterations.

The following laboratory markers presented correlations withparasite load in at least one tissue (Fig. 1): WBC (positive), PCV(negative), MCV (negative), RDW (positive), neutrophil (positive),lymphocytes (negative), eosinophil (negative), triglycerides (neg-ative), uric acid (positive), albumin (negative), total bilirrubin(positive), AST (positive), creatinine (negative), alkaline phos-phatase (positive), TAC (positive), TOC (negative), oxidative index(negative) and TBARS plasma (negative). Parasite load in total bloodwas not correlated with any given analyzed marker (see Supple-mentary Table 3).

Correlations between results for CPLD and individual tissueswere 0.972, 0.936, 0.773, 0.543 and 0.251 for left conjunctiva, rightconjunctiva, skin, buffy coat and blood, respectively. The com-pressed variable was able to capture the majority of the significantand suggestive correlations exhibited by the individual tissues, andproperly conserved the magnitude and direction of the correlationcoefficients.

4. Discussion

To date, no existing report in the literature has yet assessed thesimultaneous correlation of peripheral parasite load in multipletissues with severity of clinical signs or variation in biochemi-cal, hematological and, especially, oxidative stress markers in dogsnaturally infected with Leishmania spp. Elucidation of these corre-lations may allow for accurate identification of animals with hightransmitting ability based on routine clinical examination and labo-ratory analysis. Here, we present such an investigation, which wasfacilitated by the development of a new approach, namely CPLD,which summarizes PL data across tissues into a single variable thatserves as a proxy for net parasite infection. For the sake of clarity,we use PL, CPLD and net peripheral parasite infection interchange-ably throughout the discussion, as CPLD was shown to efficientlycapture the majority of the tissue-specific PL correlations.

The main clinical alterations observed in our study are con-sistent with those commonly found in CVL (Manna et al., 2009).We found that elevation of PL increases severity of clinical signsin infected dogs, consistent with previous reports suggestingthat increased parasite density is associated with the sever-ity of CVL (Manna et al., 2009; Reis et al., 2006). Interestingly,ophthalmic alterations were associated only with PL in conjunc-tival tissues and buffy coat, suggesting that this clinical sign isa specific indicator of local parasitism. Altogether, these resultsindicate that high peripheral parasitism can be inferred in dogspresenting clinical alterations such as alopecia, onychogryphosis,popliteal lymphadenomegaly, mucosa paleness and emaciation.Although these relationships are often speculated by veterinary

Preventive Veterinary Medicine 132 (2016) 83–87

Contents lists available at ScienceDirect

Preventive Veterinary Medicine

j o ur na l ho me page: www.elsev ier .com/ locate /prevetmed

Original article

Correlations between peripheral parasite load and common clinicaland laboratory alterations in dogs with visceral leishmaniasis

R.B.P. Torrecilhaa,∗, Y.T. Utsunomiyab, A.M. Boscoa, B.F. Almeidaa, P.P. Pereiraa,L.G. Narcisoa, D.C.M. Pereirac, L. Baptistiolli a, L. Calvo-Badod, O. Courtenayd, C.M. Nunesc,P.C. Ciarlinia,∗

a Departamento de Clínica, Cirurgia e Reproduc ão Animal, Faculdade de Medicina Veterinária de Arac atuba, UNESP-Univ Estadual Paulista, Rua ClóvisPestana 793, Dona Amélia, Arac atuba, SP, 16050-680, Brazilb Departamento de Medicina Veterinária Preventiva e Reproduc ão Animal, Faculdade de Ciências Agrárias e Veterinárias, UNESP- Univ Estadual Paulista,Via de Acesso Prof. Paulo Donato Castellane s/n, Jaboticabal, SP, 14884-900, Brazilc Departamento de Apoio, Produc ão e Saúde Animal, Faculdade de Medicina Veterinária de Arac atuba, UNESP- Univ Estadual Paulista, Rua Clóvis Pestana793, Dona Amélia, Arac atuba, SP, 16050-680, Brazild School of Life Sciences and Warwick Infectious Disease and Epidemiology Research Group (WIDER), Gibbett Hill Road, CV47AL, Coventry, England, UK

a r t i c l e i n f o

Article history:Received 20 June 2016Received in revised form 22 August 2016Accepted 23 August 2016

Keywords:Canis lupus familiarisLeishmania sppHematologyBiochemistryOxidative stressPrincipal components analysis

a b s t r a c t

Intensity of peripheral parasite infection has an important role in the transmission of Leishmania spp. fromone host to another. As parasite load quantification is still an expensive procedure to be used routinely inepidemiological surveillance, the use of surrogate predictors may be an important asset in the identifi-cation of dogs with high transmitting ability. The present study examined whether common clinical andlaboratory alterations can serve as predictors of peripheral parasitism in dogs naturally infected withLeishmania spp. Thirty-seven dogs were examined in order to establish correlations between parasiteload (PL) in multiple peripheral tissues and common clinical and laboratory findings in canine visceralleishmaniasis (CVL). Quantitative polymerase chain reaction was employed to determine PL in conjunc-tival swabs, ear skin, peripheral blood and buffy coat. Additionally, a series of hematological, biochemicaland oxidative stress markers were quantified. Correlations between net peripheral infection and severityof clinical alterations and variation in laboratory parameters were assessed through a new analyticalapproach, namely Compressed Parasite Load Data (CPLD), which uses dimension reduction techniquesfrom multivariate statistics to summarize PL across tissues into a single variable. The analysis revealedthat elevation in PL is positively correlated with severity of clinical sings commonly observed in CVL, suchas skin lesions, ophthalmic alterations, onycogriphosis, popliteal lymphadenomegaly and low body mass.Furthermore, increase in PL was found to be followed by intensification of non-regenerative anemia, neu-trophilia, eosinopenia, hepatic injury and oxidative imbalance. These results suggest that routinely usedclinical and laboratory exams can be predictive of intensity of peripheral parasite infection, which has animportant implication in the identification of dogs with high transmitting ability.

© 2016 Elsevier B.V. All rights reserved.

1. Introduction

Visceral leishmaniasis is an anthropozoonosis caused by para-sites of the Leishmania genus (Solano-Gallego et al., 2011). As theparasite is transmitted from one host to another by the bites of Lut-zomiya longipalpis sandflies, the intensity of peripheral parasitismis an important contributor to the transmitting ability of the host.

∗ Corresponding authors.E-mail addresses: [email protected] (R.B.P. Torrecilha),

[email protected] (P.C. Ciarlini).

The main urban reservoir of the disease is the domestic dog (Canislupus familiaris), and the presence of infected dogs in the vicin-ity of humans has been incriminated as a risk factor for humanleishmaniasis (Solano-Gallego et al., 2009).

Clinical staging in canine visceral leishmaniasis (CVL) has beenshown to be associated with parasite density in specific tissue com-partments, such as skin (de Almeida Ferreira et al., 2012), lymphnodes (Reale et al., 1999), spleen (Solcà et al., 2012) and bonemarrow (Reis et al., 2006). Leishmania spp. infection has also beenfound to alter markers of hematological, biochemical and oxidativehomeostasis in dogs (Almeida et al., 2013b; Nicolato et al., 2013;Reis et al., 2006). However, it remains unclear at what extent the

http://dx.doi.org/10.1016/j.prevetmed.2016.08.0060167-5877/© 2016 Elsevier B.V. All rights reserved.

CPLD – Compressed Parasite Load data




R.B.P. Torrecilha et al. / Preventive Veterinary Medicine 132 (2016) 83–87 85

Fig. 1. Correlogram of Spearman’s rank correlation coefficients between Leishmania spp. density (horizontal axis) in different tissues and clinical alterations, blood cell count,biochemical and oxidative stress markers (vertical axis). Warm colors (towards red) represent positive correlations (r > 0), whereas cool colors (towards blue) represent nega-tive correlations (r < 0). Only markers presenting at least one suggestive (0.05 ≤ P < 0.10) or significant (P < 0.05) correlation in a randomization test with 10,000 permutationsare shown. For a complete summary of correlations see the Supplementary information (Supplementary Table 3 and 4). (For interpretation of the references to colour in thisfigure legend, the reader is referred to the web version of this article.)

and skin samples, respectively. However, all dogs presented para-site infection in at least one tissue, confirming that all dogs wereinfected. Parasite load across tissues were moderately to highlycorrelated, with an average correlation of r = 0.400 (Supplemen-tary information). An exception was observed for blood, which waspoorly correlated with all tissues (average correlation of r = 0.069).

In order to have a representation of all tissues simultaneously,we created a new compressed variable, namely CPLD, based on aprincipal component analysis of the PL matrix. For that matter, werequired complete observations of PL across tissues, which reducedthe data from 37 to 31 samples. The first leading principal compo-nent explained 49.1% of the total variance contained in the originalmatrix, and therefore was adopted as the CPLD. The relative con-tributions of each tissue to the CPLD were 22.7% for skin, 28.5% forright conjunctiva, 29.6% for left conjunctiva, 12.3% for buffy coatand 6.9% for blood.

3.4. Correlations for peripheral parasite load

As the permutation test is conservative, we decided to discusssuggestive (0.05 ≤ P < 0.10) and significant (P < 0.05) correlations.All clinical alterations were positively correlated (P < 0.05) withparasite load in two or more tissues (Fig. 1). Ophthalmic alterationswere only associated with PL in the conjunctivas. Parasite load inthe blood did not present associations with clinical alterations. TheCPLD variable successfully captured the correlatons of the individ-ual tissues, and only failed to predict ophthalmic alterations.

The following laboratory markers presented correlations withparasite load in at least one tissue (Fig. 1): WBC (positive), PCV(negative), MCV (negative), RDW (positive), neutrophil (positive),lymphocytes (negative), eosinophil (negative), triglycerides (neg-ative), uric acid (positive), albumin (negative), total bilirrubin(positive), AST (positive), creatinine (negative), alkaline phos-phatase (positive), TAC (positive), TOC (negative), oxidative index(negative) and TBARS plasma (negative). Parasite load in total bloodwas not correlated with any given analyzed marker (see Supple-mentary Table 3).

Correlations between results for CPLD and individual tissueswere 0.972, 0.936, 0.773, 0.543 and 0.251 for left conjunctiva, rightconjunctiva, skin, buffy coat and blood, respectively. The com-pressed variable was able to capture the majority of the significantand suggestive correlations exhibited by the individual tissues, andproperly conserved the magnitude and direction of the correlationcoefficients.

4. Discussion

To date, no existing report in the literature has yet assessed thesimultaneous correlation of peripheral parasite load in multipletissues with severity of clinical signs or variation in biochemi-cal, hematological and, especially, oxidative stress markers in dogsnaturally infected with Leishmania spp. Elucidation of these corre-lations may allow for accurate identification of animals with hightransmitting ability based on routine clinical examination and labo-ratory analysis. Here, we present such an investigation, which wasfacilitated by the development of a new approach, namely CPLD,which summarizes PL data across tissues into a single variable thatserves as a proxy for net parasite infection. For the sake of clarity,we use PL, CPLD and net peripheral parasite infection interchange-ably throughout the discussion, as CPLD was shown to efficientlycapture the majority of the tissue-specific PL correlations.

The main clinical alterations observed in our study are con-sistent with those commonly found in CVL (Manna et al., 2009).We found that elevation of PL increases severity of clinical signsin infected dogs, consistent with previous reports suggestingthat increased parasite density is associated with the sever-ity of CVL (Manna et al., 2009; Reis et al., 2006). Interestingly,ophthalmic alterations were associated only with PL in conjunc-tival tissues and buffy coat, suggesting that this clinical sign isa specific indicator of local parasitism. Altogether, these resultsindicate that high peripheral parasitism can be inferred in dogspresenting clinical alterations such as alopecia, onychogryphosis,popliteal lymphadenomegaly, mucosa paleness and emaciation.Although these relationships are often speculated by veterinary

Preventive Veterinary Medicine 132 (2016) 83–87

Contents lists available at ScienceDirect

Preventive Veterinary Medicine

j o ur na l ho me page: www.elsev ier .com/ locate /prevetmed

Original article

Correlations between peripheral parasite load and common clinicaland laboratory alterations in dogs with visceral leishmaniasis

R.B.P. Torrecilhaa,∗, Y.T. Utsunomiyab, A.M. Boscoa, B.F. Almeidaa, P.P. Pereiraa,L.G. Narcisoa, D.C.M. Pereirac, L. Baptistiolli a, L. Calvo-Badod, O. Courtenayd, C.M. Nunesc,P.C. Ciarlinia,∗

a Departamento de Clínica, Cirurgia e Reproduc ão Animal, Faculdade de Medicina Veterinária de Arac atuba, UNESP-Univ Estadual Paulista, Rua ClóvisPestana 793, Dona Amélia, Arac atuba, SP, 16050-680, Brazilb Departamento de Medicina Veterinária Preventiva e Reproduc ão Animal, Faculdade de Ciências Agrárias e Veterinárias, UNESP- Univ Estadual Paulista,Via de Acesso Prof. Paulo Donato Castellane s/n, Jaboticabal, SP, 14884-900, Brazilc Departamento de Apoio, Produc ão e Saúde Animal, Faculdade de Medicina Veterinária de Arac atuba, UNESP- Univ Estadual Paulista, Rua Clóvis Pestana793, Dona Amélia, Arac atuba, SP, 16050-680, Brazild School of Life Sciences and Warwick Infectious Disease and Epidemiology Research Group (WIDER), Gibbett Hill Road, CV47AL, Coventry, England, UK

a r t i c l e i n f o

Article history:Received 20 June 2016Received in revised form 22 August 2016Accepted 23 August 2016

Keywords:Canis lupus familiarisLeishmania sppHematologyBiochemistryOxidative stressPrincipal components analysis

a b s t r a c t

Intensity of peripheral parasite infection has an important role in the transmission of Leishmania spp. fromone host to another. As parasite load quantification is still an expensive procedure to be used routinely inepidemiological surveillance, the use of surrogate predictors may be an important asset in the identifi-cation of dogs with high transmitting ability. The present study examined whether common clinical andlaboratory alterations can serve as predictors of peripheral parasitism in dogs naturally infected withLeishmania spp. Thirty-seven dogs were examined in order to establish correlations between parasiteload (PL) in multiple peripheral tissues and common clinical and laboratory findings in canine visceralleishmaniasis (CVL). Quantitative polymerase chain reaction was employed to determine PL in conjunc-tival swabs, ear skin, peripheral blood and buffy coat. Additionally, a series of hematological, biochemicaland oxidative stress markers were quantified. Correlations between net peripheral infection and severityof clinical alterations and variation in laboratory parameters were assessed through a new analyticalapproach, namely Compressed Parasite Load Data (CPLD), which uses dimension reduction techniquesfrom multivariate statistics to summarize PL across tissues into a single variable. The analysis revealedthat elevation in PL is positively correlated with severity of clinical sings commonly observed in CVL, suchas skin lesions, ophthalmic alterations, onycogriphosis, popliteal lymphadenomegaly and low body mass.Furthermore, increase in PL was found to be followed by intensification of non-regenerative anemia, neu-trophilia, eosinopenia, hepatic injury and oxidative imbalance. These results suggest that routinely usedclinical and laboratory exams can be predictive of intensity of peripheral parasite infection, which has animportant implication in the identification of dogs with high transmitting ability.

© 2016 Elsevier B.V. All rights reserved.

1. Introduction

Visceral leishmaniasis is an anthropozoonosis caused by para-sites of the Leishmania genus (Solano-Gallego et al., 2011). As theparasite is transmitted from one host to another by the bites of Lut-zomiya longipalpis sandflies, the intensity of peripheral parasitismis an important contributor to the transmitting ability of the host.

∗ Corresponding authors.E-mail addresses: [email protected] (R.B.P. Torrecilha),

[email protected] (P.C. Ciarlini).

The main urban reservoir of the disease is the domestic dog (Canislupus familiaris), and the presence of infected dogs in the vicin-ity of humans has been incriminated as a risk factor for humanleishmaniasis (Solano-Gallego et al., 2009).

Clinical staging in canine visceral leishmaniasis (CVL) has beenshown to be associated with parasite density in specific tissue com-partments, such as skin (de Almeida Ferreira et al., 2012), lymphnodes (Reale et al., 1999), spleen (Solcà et al., 2012) and bonemarrow (Reis et al., 2006). Leishmania spp. infection has also beenfound to alter markers of hematological, biochemical and oxidativehomeostasis in dogs (Almeida et al., 2013b; Nicolato et al., 2013;Reis et al., 2006). However, it remains unclear at what extent the

http://dx.doi.org/10.1016/j.prevetmed.2016.08.0060167-5877/© 2016 Elsevier B.V. All rights reserved.

CPLD – Compressed Parasite Load data

Variazioni ematobiochimiche in corso di vaccinazione Canileish®

•  Lieve incremento proMdemia (ns) •  Incremento C-‐Rea Prot (p <0.05) •  SieroeleYroforesi (p <0.05)

–  Incremento beta 1 e 2 globuline –  Incremento gamma globuline

•  Incremento IgG post 6-‐8 mesi (p <0.05) •  Incremento IgM e IgA post 2 mesi (p <0.05)


(Starita et al, 2016; Vannucci et al, 2017)

diagnosco terapeuco gesonaleinassenzadicrisileishmanioca ... · introduction: canine visceral...

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