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Canine leishmaniosis caused by Leishmania major and Leishmania tropica: comparative findings and serology.

ABSTRACT: Infection and clinical disease associated with Leishmania major and Leishmania tropica, two common agents of human cutaneous leishmaniosis, have rarely been reported in dogs. This study describes dogs infected with these Leishmania spp. prevalent in the Middle East and North Africa, and compares the serological response of dogs infected with Leishmania infantum, L. major or L. tropica to whole promastigote antigen enzyme-linked immunosorbent assay (ELISA) of each species and to rK39 dipstick.Leishmania major infection in a 5-month-old male dog was associated with alopecic and ulcerative periocular and limb skin lesions which responded to allopurinol treatment. Infection was detected by skin and blood polymerase chain reaction (PCR) and confirmed by DNA sequencing but the dog was seronegative. Leishmania tropica infection was detected in a 3-month-old female dog co-infected with Babesia vogeli and Anaplasma platys and with no skin lesions. PCR and DNA sequencing of the blood and parasite culture were positive for L. tropica. Sera from 11 dogs infected with L. infantum, L. major or L. tropica were reactive with all three Leishmania spp. antigens except for sera from a dog with L. major infection. No significant differences were found between reactivity of dog sera to the antigen of the infecting species, or to the other Leishmania spp. antigens. Sera from dogs infected with L. infantum and L. tropica were positive with the rK39 antigen kit, while dogs with L. major infection were seronegative.Skin lesions in L. major infected dogs from this study and previous reports (n?=?2) were ulcerative and located on the muzzle, feet and foot pads and not associated with generalized lymphadenomegaly and splenomegaly. In previous L. tropica infections, skin lesions were proliferative mucocutaneous in young dogs (n?=?2), or associated with widespread dermatitis, lymphadenomegaly and splenomegaly in older dogs with similarity to L. infantum infection (n?=?2). This study suggests that ELISA serology with whole promastigote antigen is not distinctive between L. infantum, L. major and L. tropica canine infections and that some L. major infections are not seropositive. PCR with DNA sequencing should be used to discriminate between canine infections with these three species.

SUBMITTER: Baneth G

PROVIDER: S-EPMC5346844 | biostudies-literature | 2017 Mar

REPOSITORIES: biostudies-literature

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Canine leishmaniosis caused by Leishmania major and Leishmania tropica: comparative findings and serology.

Baneth Gad G Yasur-Landau Daniel D Gilad Matan M Nachum-Biala Yaarit Y

Parasites & vectors 20170313 1

<h4>Background</h4>Infection and clinical disease associated with Leishmania major and Leishmania tropica, two common agents of human cutaneous leishmaniosis, have rarely been reported in dogs. This study describes dogs infected with these Leishmania spp. prevalent in the Middle East and North Africa, and compares the serological response of dogs infected with Leishmania infantum, L. major or L. tropica to whole promastigote antigen enzyme-linked immunosorbent assay (ELISA) of each species and t ...[more]

PMID: 28285601

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Project description:Cutaneous Leishmaniasis (CL) is a parasitic infection classified by the WHO as one of the most uncontrolled spreading neglected diseases. Syria is endemic for Leishmania tropica and Leishmania major, causing CL in the Eastern Mediterranean. The large-scale displacement of Syrian refugees exacerbated the spread of CL into neighboring countries. Therapeutic interventions against CL include local, systemic and physical treatments. The high risk for drug-resistance to current treatments stresses the need for new therapies. Imiquimod is an immunomodulatory drug with a tested efficacy against L. major species. Yet, Imiquimod efficacy against L. tropica and the molecular mechanisms dictating its potency are still underexplored. In this study, we characterized the effect of Imiquimod against L. tropica and L. major, and characterized the molecular mechanisms dictating its anti-leishmanial efficacy against both strains. We also investigated the potency and molecular mechanisms of an Imiquimod analog, EAPB0503, against these two strains. We have tested the effect of Imiquimod and EAPB0503 on macrophages infected with either L. major, L. tropica strains, or patient-derived freshly isolated L. tropica parasites. The anti-amastigote activity of either drugs was assessed by quantitative real time PCR (RT-PCR) using kinetoplast specific primers, confocal microscopy using the Glycoprotein 63 (Gp63) Leishmania amastigote antibody or by histology staining. The mechanism of action of either drugs on the canonical nuclear factor kappa- B (NF-?B) pathway was determined by western blot, and confocal microscopy. The immune production of cytokines upon treatment of infected macrophages with either drugs was assessed by ELISA. Both drugs reduced amastigote replication. EAPB0503 proved more potent, particularly on the wild type L. tropica amastigotes. Toll-Like Receptor-7 was upregulated, mainly by Imiquimod, and to a lesser extent by EAPB0503. Both drugs activated the NF-?B canonical pathway triggering an immune response and i-NOS upregulation in infected macrophages. Our findings establish Imiquimod as a strong candidate for treating L. tropica and show the higher potency of its analog EAPB0503 against CL.

Project description:BackgroundLeishmaniosis, a vector-borne disease caused by Leishmania infantum, is one of the most important parasitic zoonoses in Europe. The transmission cycle of leishmaniosis is maintained by both domestic and wild animals. However, few data are available on the role of wild mammals in transmitting the parasite in the European Mediterranean basin. As feline leishmaniosis, diagnosis of the infection in ferrets can be a challenge, the use of different serological and molecular methods combined is a recommended approach. Our aim was to investigate the prevalence of infection of L. infantum in apparently healthy domestic ferrets (Mustela putorius furo) in an endemic region of Spain (Community of Valencia), using serological and molecular methods and to evaluate the results comparing the different techniques.MethodsThe prevalence of Leishmania infection was studied in domestic ferrets. Blood was collected from each animal for serology and molecular analysis. Two serological methods, enzyme-linked immunosorbent assay (ELISA) and western blot (WB), were used for the detection of L. infantum antibodies, and real-time polymerase chain reaction (qPCR) was used for the detection of L. infantum DNA.ResultsBlood samples from 102 apparently healthy ferrets were analyzed. In the serological study, 25.5% of the animals tested positive by western blot, and 9.0% by enzyme-linked immunosorbent assays. The seroprevalence of L. infantum infection, based on a positive result in any serological test, was 28.4% (95% confidence interval [CI] 20.6-S37.9%). No kinetoplast DNA (kDNA) was detected by qPCR in peripheral blood samples from the ferrets tested.ConclusionsThe immunological response revealed by these tests indicates that the ferrets are exposed to repeated inoculations with the endemic parasite L. infantum. Although the low population of domestic ferrets means their reservoir potential is limited in the absence of a primary host, it would be of interest to carry out further studies using xenodiagnosis to determine whether they are accidental or reservoir host species capable of spreading infection.

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Project description:BackgroundLeishmaniosis, zoonosis that produces significant public health impacts, is caused by Leishmania infantum. Canines are the main domestic reservoir and, besides humans, other species of mammals could be infected when living in endemic areas. In this study, we detected equine Leishmania infantum infections in a canine visceral leishmaniosis transmission area and evaluated the clinical, haematological, biochemical and oxidative stress disorders. This study was conducted in Uruguaiana, Rio Grande do Sul, south of Brazil. Peripheral blood samples were collected from 124 animals (98 horses and 26 dogs) of both genders and several breeds after they underwent general and dermatologic examinations.ResultsTwenty five Leishmania infantum infected animals (20.16%), 14 horses and 11 dogs were detected by PCR (Polymerase Chain Reaction) amplification of kinetoplast DNA regions with 96% homology to Leishmania infantum (GenBank Accession No. L 19877.1). The clinical and haematological alterations of infected equines were skin lesions, nodules, lymphadenopathy, decreased levels in red blood cells and haematocrit (p < 0.05) and increase in urea serum concentration (p < 0.05), while CVL presented a decrease in red blood cells counts (p < 0.05), increase in lymphocytes (p < 0.05), and decrease in neutrophil-lymphocyte ratio (p < 0.05). Oxidative stress markers of plasma protein carbonyl and plasma lipid peroxidation were not statistically significant (p > 0.05) in both species.ConclusionsTo our knowledge, this has been the first leishmaniosis equine survey performed in south of Brazil, caused by Leishmania infantum that were able to initially identify haematological and biochemical changes in the species, even in asymptomatic animals. We present evidence supporting those findings of haematological and biochemical changes could be related to infection. Surprisingly, the clinical manifestations of equine infection were similar to those found in canine visceral leishmaniosis. The equine population could be play an important role in the cycle of leishmaniosis in south Brazil and consequently indicates a great risk of public health. This evaluation of infected animals is important to establish the clinical and laboratory parameters involved in the disease progression.

Dataset Information

Canine leishmaniosis caused by Leishmania major and Leishmania tropica: comparative findings and serology.

Publications

Canine leishmaniosis caused by Leishmania major and Leishmania tropica: comparative findings and serology.

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