scholarly journals Serological and molecular diagnostic tests for canine visceral leishmaniasis in Brazilian endemic area: one out of five seronegative dogs are infected

2017 ◽  
Vol 145 (12) ◽  
pp. 2436-2444 ◽  
Author(s):  
E. G. LOPES ◽  
A. P. SEVÁ ◽  
F. FERREIRA ◽  
C. M. NUNES ◽  
L. B. KEID ◽  
...  

SUMMARYEuthanasia of infected dogs is one of the measures adopted in Brazil to control visceral leishmaniasis (VL) in endemic areas. To detect infected dogs, animals are screened with the rapid test DPP® Visceral Canine Leishmaniasis for detection of antibodies against K26/K39 fusion antigens of amastigotes (DPP). DPP-positives are confirmed with an immunoenzymatic assay probing soluble antigens of promastigotes (ELISA), while DPP-negatives are considered free of infection. Here, 975 dogs from an endemic region were surveyed by using DPP, ELISA and real-time PCR (qPCR) for the diagnosis of VL. When DPP-negative dogs were tested by qPCR applied in blood and lymph node aspirates, 174/887 (19·6%) were positive in at least one sample. In a second sampling using 115 cases, the DPP-negative dogs were tested by qPCR in blood, lymph node and conjunctival swab samples, and 36/79 (45·6%) were positive in at least one sample. Low-to-moderate pairwise agreement was observed between all possible pair of tests. In conclusion, the official diagnosis of VL in dogs in Brazilian endemic areas failed to accuse an expressive number of infected animals and the impact of the low accuracy of serological tests in the success of euthanasia-based measure for VL control need to be assessed.

2021 ◽  
Vol 49 ◽  
Author(s):  
Gilsan Aparecida De Oliveira ◽  
Vitória Aline Santos Sarmento ◽  
Isabelle Vanderlei Martins Bastos ◽  
Alberon Ribeiro De Araújo ◽  
Lígia Buzzá Roo De Mendonça ◽  
...  

Background: Canine visceral leishmaniasis (CVL) is a parasitic disease of high lethality caused by the protozoan Leishmania infantum in Brazil and is often related to splenomegaly. However, splenic nodules in dogs, although frequent, have not previously been reported as associated with CVL, but with neoplastic diseases. Considering that most dogs infected are oligosymptomatic or asymptomatic and that splenic nodules are common to other diseases, it is prudent to differentially diagnose CVL in view of its high zoonotic potential and lethality. The objective of the study was to describe a case of splenomegaly with splenic nodules associated with CVL in an asymptomatic dog treated with 2% miltefosina.                                             Case: A 5-year-old male Rottweiler with 41 kg, with a history of inappetence, apathy and weight loss was referred to the Veterinary Medicine School Clinic of the Cesmac University Center, Maceió, AL, Brazil. However, during palpation a slight increase in the spleen was noted. Hematological, hemoparasite, biochemical and abdominal ultrasonographic examinations were requested to clarify the clinical suspicion of hemoparasitosis. The hematological and biochemical results respectively showed the following: normocytic normochromic anemia, hyperproteinemia and thrombocytopenia, in addition to hypoalbuminemia, with elevated total protein levels. The test for hemoparasites was negative. Ultrasonography showed mixed echogenicity suggestive of nodules. The rapid test for Ehrlichia, Anaplasma and L. infantum was performed. It was positive only for L. infantum. ELISA, IFAT and qPCR tests were performed to confirm the result. The test showed a cut-off result of 0.371 for ELISA, positive for RIFI at a cut-off of 1:40 and qPCR with less than 1 fg and with amplification above 36 cycles. In view of these results, treatment with 2% miltefosine at a dose of 1 mL/ 10 kg was started once a day, after feeding, for 28 days. The animal was monitored throughout treatment and re-evaluated every 10 days for 30 days, showing signs of clinical development, presenting satisfactory results.Discussion: Canine splenomegaly can be associated with a variety of disease possibilities. In asymptomatic canine visceral leishmaniasis (CanL), the slight increase in spleen and the presence of splenic nodules may lead to a false diagnosis. Splenic nodules may be associated with dogs of advanced age and may be due to lymphoid nodular hyperplasia, which causes nodules with echogenicity, hyperechoic regions with well demarcated irregularity, with centralized hypoechoic areas and an absence of hematological and biochemical alterations. The cause of splenomegaly associated with nodules may be difficult to diagnose and require much time and effort. Therefore, diseases such as visceral leishmaniasis of high lethality must be the priority in differential diagnosis in endemic areas in order to minimize the risk of transmission. In addition to allowing an early intervention aiming at good animal health results and preventive measures, such as the use of repellent collars that reduce the risk of phlebotomo infection. The differential diagnosis of CVL is necessary in endemic areas, even in asymptomatic dogs that may present splenic alterations suggestive of other diseases. Treatment with 2% miltefosine was shown to be, in this case, effective at reducing the splenic nodules and a good alternative for the quality of life of the animal.


2019 ◽  
Vol 4 (Suppl 3) ◽  
pp. A36.1-A36
Author(s):  
Salah Boshara

BackgroundVisceral leishmaniasis (VL) also known as kala-azar, is a protozoan infection caused by the L. donovani complex and transmitted by sandflies. Early detection of leishmaniasis is critical in management of patients and for successful control and elimination of the disease. Definitive diagnosis of visceral leishmaniasis is by parasitological demonstration of parasites in splenic, lymph node or bone marrow aspirates, which are collected using invasive methods that are unsuitable in the field. This study aimed to evaluate new less invasive urine-based ELISA and rapid diagnostic test (RDT) assays for diagnosis of VL.MethodsThe newly developed urine ELISA test was evaluated using archived and fresh urine samples collected from parasitologically confirmed VL patients and non-VL cases. Lateral flow assay (LFA) using the ELISA reagents were conducted for day0 samples. Serological tests (DAT, rk28 ICT) were conducted for every patient in the study.ResultsIn 198 patients with suspected VL, urine rapid test had a sensitivity of 72.2% and exhibited a specificity of 93.42%. Leishmania antigen ELISA had a sensitivity of 83.33% and a specificity of 95.05%. All VL-confirmed cases were followed up during the treatment period, the Leishmania antigen ELISA became negative 2 months after completion of treatment in most patients.ConclusionUrine lateral flow assay is a simple addition to the diagnostics of VL particularly at field level and as a complementary test for the diagnosis of VL in smear-negative cases. Further enhancement of the test will define its performance in monitoring treatment. Further studies are recommended to evaluate the performance of both tests in the diagnosis of HIV-co-infected cases.


Author(s):  
Asier Basurco ◽  
Alda Natale ◽  
Katia Capello ◽  
Antonio Fernández ◽  
María Teresa Verde ◽  
...  

Abstract Canine leishmaniasis (CanL) is a disease caused by Leishmania infantum. Serological methods are the most common diagnostic techniques used for the diagnosis of the CanL. The objective of our study was to estimate the sensitivity and specificity of one in-house ELISA kit (ELISA UNIZAR) and three commercially available serological tests (MEGACOR Diagnostik GmbH) including an immunochromatographic rapid test (FASTest LEISH®), an immunofluorescent antibody test (MegaFLUO LEISH®) and an enzyme-linked immunosorbent assay (MegaELISA LEISH®), using latent class models in a Bayesian analysis. Two hundred fifteen serum samples were included. The highest sensitivity was achieved for FASTest LEISH® (99.38%), ELISA UNIZAR (99.37%), MegaFLUO LEISH® (99.36%) followed by MegaELISA LEISH® (98.49%). The best specificity was obtained by FASTest LEISH® (98.43%), followed by ELISA UNIZAR (97.50%), whilst MegaFLUO LEISH® and MegaELISA LEISH® obtained the lower specificity (91.94% and 91.93%, respectively). The results of present study indicate that the immunochromatographic rapid test evaluated FASTest LEISH® show similar levels of sensitivity and specificity to the quantitative commercial tests. Among quantitative serological tests, sensitivity and specificity were similar considering ELISA or IFAT techniques.


2021 ◽  
Vol 22 ◽  
Author(s):  
Aline Oliveira de Magalhães ◽  
Lucilandia Maria Bezerra ◽  
Diego Pereira Araújo ◽  
Bruna Siqueira Gomes de Lima ◽  
Leandro do Padro Assunção ◽  
...  

Abstract Canine leishmaniasis (CanL) is a zoonosis caused by the protozoan of the species Leishmania infantum. The spleen and lymph nodes undergo morphological changes during CanL. This research aimed to perform an anatomopathological and immunohistochemical study of these organs in dogs reactive to leishmaniasis in the Dual-path Platform chromatographic immunoassay (DPP®) and Enzyme Immunoabsorption Assay (ELISA). Twenty-seven dogs were evaluated for anatomopathological examination with 92.6% showing changes at gross evaluation, specially splenomegaly and lymphadenomegaly. All dogs showed changes in the spleen unrelated to the parasitic load, with granulomatous splenitis being the most severe change. Diffuse cortical and paracortical hyperplasia, and hyperplasia and hypertrophy of the medullary cords were observed in the lymph node. Amastigote forms of Leishmania spp. were found in the spleen and lymph node at histopathological and immunohistochemical evaluations, with good agreement between these evaluations (k = 0.55, p = 0.00124), but no difference was observed in the parasitic intensity of these organs at immunohistochemistry (p = 0.23). It was concluded that spleen and lymph node from dogs reactive to leishmaniasis on the DPP® and ELISA tests show histomorphological changes resulting from the disease, independent to the parasitic load, as well as these organs show similar parasitic load at immunohistochemical test.


2020 ◽  
Author(s):  
Ana Isabel Olías-Molero ◽  
Erica Fontán-Matilla ◽  
José María Alunda

Abstract Background Zoonotic visceral leishmaniasis by Leishmania infantum is a first order pathology in veterinary clinics of dogs in endemic areas. Moreover, canine infections (CanL) are considered the main reservoir for human disease. Despite the importance of CanL in the control of the diseases within a One Health approach no scientometric study has been published. Aims of the study included the impact of CanL on scientific literature, drugs or combinations used, trends in the period from 2000-2017 and efficacy criteria employed. Methods A Web of Science (WOS) based analysis of publications on CanL and chemotherapy of the disease in the period 2000-2017 was carried out using a stepwise methodology. Data were analyzed by year, geographical origin, chemical groups and drugs and efficacy criteria. Results Reports on CanL represented a 20% of all publications on leishmaniasis and from these ca. 14% were related to chemotherapy. Publications records on CanL followed the distribution of the infection in endemic areas although an overrepresentation of Mediterranean countries was evident. Published reports on the main antileishmanial drugs used in clinical practice showed a sustained tendency in the period analyzed. Sb V , alone or in combination with allopurinol, represented over 50% of all publications on chemotherapy of CanL despite the availability of more recently marketed drugs. Conclusions Chemotherapy of CanL still relies on Sb V and combinations. Reports on chemotherapy are scarce, mostly publicly funded and the variability of experimental conditions make extraordinarily difficult the comparison of efficacy. The vast majority of reports on efficacy do not include any information on supportive therapy; this reduces the actual value of the experiments if intended for the practical management of the disease. Complete reports of the treatments (etiological + symptomatic) would add value to the assays performed.


2020 ◽  
Vol 8 (12) ◽  
pp. 1847
Author(s):  
Margherita Ortalli ◽  
Daniele Lorrai ◽  
Paolo Gaibani ◽  
Giada Rossini ◽  
Caterina Vocale ◽  
...  

This study compares the performance of seven assays, including two ELISA (Leishmania ELISA IgG + IgM, Vircell Microbiologists; Leishmania infantum IgG ELISA, NovaTec), three rK39-based immunochromatographic tests (rK39-ICTs) (Leishmania Dipstick Rapydtest, Apacor; On Site Leishmania IgG/IgM Combo Rapid Test, CTK Biotech; LEISHMANIA Strip quick Test, Cypress Diagnostic), one indirect immunofluorescent antibody test (IFAT) (Leishmania-Spot IF, BioMérieux), and one western blot (WB) (Leishmania WESTERN BLOT IgG, LDBio Diagnostics) for serodiagnosis of visceral leishmaniasis (VL). Serum samples from 27 VL patients living in northeastern Italy were analyzed, as well as the serum samples from 50 individuals in whom VL diagnosis was excluded. The WB and the IFAT had 96% sensitivity, followed by the ELISA (63% and 74%, respectively). The rK39-ICT exhibited the worst performance among the serological tests, with sensitivities ranging from 52% to 70%. By combining selected ELISA/ICT, the sensitivity of VL detection reached 89%. IFAT and WB outperformed ELISA and rK39-ICT by possessing optimal sensitivity, but their high cost and complexity of execution would not allow their employment as screening tests. In conclusion, the combination of easy-to-perform tests, such as ICT and ELISA, could improve sensitivity in the serodiagnosis of Mediterranean VL.


2018 ◽  
Vol 44 (1) ◽  
pp. 6
Author(s):  
Mahyumi Fujimori ◽  
Arleana Do Bom Parto Ferreira de Almeida ◽  
Álvaro Felipe Lima Ruy Dias ◽  
Juliana Yuki Rodrigues ◽  
Luciano Nakazato ◽  
...  

Background: Canine visceral leishmaniasis is a chronic and severe disease of great interest to global public health. In Brazil, the main species causing visceral leishmaniasis is Leishmania chagasi [syn. Leishmania infantum], which is transmitted by sandflies of the species Lutzomyia longipalpis and Lu. cruzi. This study aimed to determine the risk factors and prevalence of visceral leishmaniasis in dogs residing in endemic areas of the municipality of Várzea Grande, Mato Grosso.Material, Methods & Results: The study was conducted in the Várzea Grande neighbourhoods Jardim Eldorado, Parque Sabiá and São Mateus, which are located in the Brazilian Cerrado region and are endemic for visceral leishmaniasis. The Várzea Grande is a mostly commercial and industrial municipality and relies on subsistence farming. To describe the general characteristics of the dog population and housing environment, an interview was carried out with the dog owners in each household. This interview addressed local demographics and the identification of dogs to establish epidemiological aspects of canine illness and risk factors for infection. The dogs were examined and physically restrained for blood collection by jugular venipuncture. For serological testing, a commercial enzyme-linked immunosorbent assay (ELISA) was used. A Chi-square test or Fisher’s exact test was used to identify associations between independent variables and the seroprevalence of dogs with anti-Leishmania antibodies. It were collected blood from 521 animals, including 160 belonging to Jardim Eldorado, 129 to Parque Sabiá and 232 to São Mateus. Of these animals, 120 were reactive by ELISA, with a prevalence of 23.0%. There was no statistically significant difference (P > 0.05) according to gender, age, racial definition, origin, time with family, presence of ectoparasites, or the presence of people with skin wounds at home or on other pets. The major risk factor for canine infection was fur length; dogs with short fur were 2.2 times more likely to be infected than dogs with long fur.Discussion: The prevalence of canine leishmaniasis in endemic areas was found to be similar to studies in Cuiaba, a city in Várzea Grande, which also included neighbourhoods endemic for visceral leishmaniasis as study areas. Other studies have shown prevalence rates that differ from those found in this study. This difference may either be because the neighbourhoods in this study had already experienced outbreaks of the disease or because serum samples were cross-reactive with other trypanosomatids. While infection was more readily detected in dogs with clinical signs, asymptomatic dogs with anti-Leishmania antibodies were found in greater numbers, which is consistent with the literature. Hair length was the only factor associated with canine leishmaniasis in this study; short-haired animals were most at risk of contracting the disease because they had more skin exposure to sandflies during blood feeding. In conclusion, endemic districts of Várzea Grande had high prevalence rates of canine visceral leishmaniasis, and a short length coat was associated with infection.


2016 ◽  
Vol 44 (1) ◽  
pp. 4
Author(s):  
Matheus Viezzer Bianchi ◽  
Gabriela Fredo ◽  
Nelson Junior Tagliari ◽  
Ronaldo Viana Leite Filho ◽  
Cintia De Lorenzo ◽  
...  

Background: Leishmaniasis is a chronic infectious disease caused by intracellular protozoan Leishmania that affects canine and human. The visceral leishmaniasis (VL) is caused by the Leishmania donovani complex, in which canines are the main reservoir for human VL. In Southern Brazil, case reports of this disease have increased, especially when canines are infected in endemic areas in the country. Canines usually present a systemic disease, characterized by cutaneous lesions, weight loss, generalized lymphadenopathy, hepatomegaly and splenomegaly. This report aims to describe the clinicalpathological features of a case of autochthonous VL in a canine of an urban area of Porto Alegre.Case: A male, mixed breed, dog , 7 year-old, resident at the East Side of Porto Alegre, living together with two other canines, presented at clinical examination progressive weight loss, associated with hyporexia and hyperthermia, evolving to lateral recumbency, paralisys, and death. At necropsy, the dog was cachectic with diffusely pale mucosae. Gross findings included liver enlarged, with multifocal firm brownish areas, spleen enlarged, showing multifocal firm round dark-red areas, and kidneys diffusely pale with evidentiated tranversal striations on cortical zone. At microscopic examination, there was on the spleen diffuse inflammatory infiltrate of macrophages with large cytoplasm containing multiple amastigotes. The liver, exhibited atrophy of hepatocytes and moderate multifocal inflammatory infiltrate in sinusoids of macrophages containing multiple amastigotes. These features were also observed moderately on lymph nodes and lamina propria of large intestine. Immunohistochemistry examination showed marked positive staining for Leishmania spp. in amastigotes located whitin the cytoplasm of macrophages of spleen, liver, lymph nodes and large intestine.Discussion: Canine leishmaniasis is a disease that affects both internal organs and skin. However, the condition is called VL mostly because it is associated with the same species that cause the visceral form in humans. In this case the canine presented liver, spleen and lymph node lesions; however no skin lesion was observed during the clinical examination. A serological evaluation of canids in East Side of Porto Alegre performed in 2005 showed that 3.5% of these dogs were positive for Leishmania; yet neither of these animals presented clinical signs. This demonstrates that the agent was already present, however only 3-10% of the canines infected develop clinical disease. In non-endemic areas VL is related to the migration of canids from endemic areas, where VL is common. However the canine here described was born, raised, and was living in Porto Alegre. Lutzomyia longipalpis is the main vector of VL and despite its wide distribution it was identified only in 2009 in Rio Grande do Sul State. Autochthonous cases of VL were reported in canines and humans in this state, confirming the hypothesis that the vector was widely distributed and it is now present in Porto Alegre, keeping the epidemiological cycle of that disease active. Canine VL occurs prior in humans, since canines are the main domestic reservoirs and are critical for the maintenance of this disease cycle. This is the first autochthonous VL in a canine of an urban area of Porto Alegre; therefore authorities should be alert, and new control measures must be taken to avoid the canine leishmaniasis outbreak due to its potential for zoonotic transmission.Keywords: leishmaniasis, canine, protozoan, immunohistochemistry.


2016 ◽  
Vol 10 (09) ◽  
pp. 895-901 ◽  
Author(s):  
Rodolfo A Kolliker-Frers ◽  
Ivan Insua ◽  
Gabriela Razzitte ◽  
Francisco Capani

Introduction: Argentina has been a preferential target for Bolivian immigrants for decades. The relatively recent migratory flux includes Germany, France, the United States, Australia, Japan, and some Latin American countries. The aim of this cross-sectional study was to describe the prevalence of Chagas disease in pregnant women, analyzing the Bolivian-specific Chagas prevalence as the main contributor of migratory populations from Chagas disease-endemic areas to Buenos Aires city, Argentina, and to evaluate the impact of these migrant influxes on the process of the "urbanization" of the disease in reference hospital José Maria Ramos Mejia (JMRM). Methodology: Overall, 21,332 pregnant women (100%) between 15 and 49 years of age derived from the public maternity service of JMRMH were studied. Serology data was obtained from registered serological diagnosis data, consisting of three different serological tests performed at the Public Parasitology Unit. Results: Although general prevalence decreased during the analyzed period, the specific prevalence of pregnant women from Bolivian origin showed a sustained growth during 1983–2013. Solely 5% of the total pregnant women population from Bolivia contributed to one third of the total Chagas prevalence. Conclusions: This study showed that a cohort of pregnant women from Bolivia who attended JMRMH during the period 1983–2007 constituted a population at risk for congenital transmission. Increased migration from endemic areas of Bolivia might potentially increase the prevalence of Chagas disease among pregnant women. In addition, this study highlights the importance to analyze specific prevalence according to endemic areas to determine the profiles of potential hidden prevalence.


2020 ◽  
Author(s):  
Ana Isabel Olías-Molero ◽  
Erica Fontán-Matilla ◽  
Montserrat Cuquerella ◽  
José María Alunda

Abstract Background: Zoonotic visceral leishmaniasis by Leishmania infantum is a first order pathology in veterinary clinics of dogs in endemic areas. Moreover, canine infections are considered the main reservoir for human disease; despite their importance in the control of the disease within a One Health approach no scientometric study has been published. Aims of the study included the impact of canine leishmaniasis (CanL) on scientific literature, drugs or combinations used, trends in the period from 2000-2020 and efficacy criteria employed. Methods: A Web of Science (WOS) based analysis of publications on CanL and chemotherapy of the disease in the period 2000-2020 was carried out using a stepwise methodology. Data were analyzed by year, geographical origin, chemical groups, drugs and combinations, and efficacy criteria. Results: Reports on CanL (n=3,324) represented less than 16% of all publications on leishmaniasis (n=20,968) and from these around 18% (n=596) were related to chemotherapy. Publication records on CanL followed the distribution of the infection by L.infantum in endemic areas although Mediterranean countries were overrepresented in the reports on chemotherapy of CanL. Publications on the main antileishmanial drugs used in clinical practice showed a sustained tendency in the period analyzed. Pentavalent antimonials (Sb V ), alone or in combination with allopurinol, represented over 50% of all publications on chemotherapy of CanL despite the availability of more recently marketed drugs. Conclusions: Chemotherapy of CanL still relies on Sb V and combinations and to a lesser extent on miltefosine (MIL). Reports on chemotherapy are scarce, mostly publicly funded and the variability of experimental conditions hampers the direct comparison of the efficacy of drugs, combinations and schedules. The vast majority of reports on efficacy do not include any information on supportive therapy; this reduces the actual value of the studies if intended for the practical management of the disease. Complete reports of the chemotherapy (etiological + symptomatic) would add value to the trials performed.


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