Serological, molecular, and microscopic detection of Leishmania in cats (Felis catus) in Belo Horizonte, Minas Gerais State, Brazil

Abstract The role of cats in the epidemiological cycle of leishmaniasis remains unclear. To better understand the occurrence of leishmaniasis in cats, we studied the frequency of Leishmania in serum samples of 100 cats living in an endemic region for canine and human leishmaniasis by serological, parasitological, and molecular methods. Of the 100 cats, 54 were seropositive for Leishmania antibodies by immunofluorescence antibody test. None of the bone marrow aspirates collected from these cats tested positive for the parasite in culture or upon polymerase chain reaction (PCR) analysis. Biopsy samples of the ears also tested negative for Leishmania upon PCR analysis. These findings may indicate that the region is endemic for canine leishmaniasis and cats are infected by Leishmania; or that cross-reaction with antibodies against other parasites increases the frequency of seropositivity; or that cats respond to Leishmania infection by producing antibodies when few or no parasites are present in bone marrow and tissue samples. Overall, our results suggest that cats can be infected by Leishmania ; however, we failed to demonstrate feline parasitosis. These findings highlight the need to study leishmaniasis in cats, since sandflies feed on cats, these animals may act as a reservoir for the parasite.

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DETECTION OF LEISHMANIA INFECTION IN STRAY DOGS IN HUMAN LEISHMANIASIS ENDEMIC AREA IN MYMENSINGH DISTRICT WITH ITS POSSIBLE PUBLIC HEALTH SIGNIFICANCE IN BANGLADESH

Journal of Veterinary Medical and One Health Research ◽

10.36111/jvmohr.2019.1(1).0009 ◽

2019 ◽

Vol 1 (1) ◽

Author(s):

M. A. Hossen

Keyword(s):

Visceral Leishmaniasis ◽

Asymptomatic Carrier ◽

Asymptomatic Infection ◽

Reservoir Host ◽

Sand Fly ◽

Leishmania Infection ◽

Endemic Region ◽

Stray Dogs ◽

Human Leishmaniasis ◽

Strip Test

Background: Leishmaniasis is primarily caused by two species of Leishmania (L. donovani and L. infantum) of which clinical infection with L. infantum has been recognized in both humans and dogs as zoonotic disease with dogs as the main reservoir hosts in the Mediterranean, the Middle East, Asia and South America. Although L. donovani has been associated with both clinical and asymptomatic infection in humans but it is still associated with asymptomatic infection in dogs in Indian sub-continent without any evidence of zoonotic infection. Objectives: The objective of this research was to investigate the potentiality of dog as reservoir host for visceral leishmaniasis in the human leishmaniasis endemic regions in Bangladesh. Materials and Methods: A total of 20 stray dogs in the human VL endemic areas of Mymensingh district were captured for the detection VL during the period of November 2010 to May 2011. The dipstick test rK39 (Bios International; n = 20), Giemsa’s stained impression smears of liver and spleen (n = 6) and PCR with the tissue of liver and spleen (n = 6) were tested as per manufacturer instructions and conventional standard methods. Results: Out of 20 stray dogs examined, 4 (20.0%) were positive for L. donovani infection with rK39 strip test. Of the six randomly selected dogs tested with Modified Giemsa’s stained of impression smears of spleen and liver showed 2 (33.33%) positive whereas PCR technique detected 5 (83.33%) positive for L. donovani. Results of PCR showed 145bp amplicon, specific for L. donovani infection in 83.33% stray dogs. Conclusions: This study reveals that a high percentage of L. donovani asymptomatic carrier infections occur in dogs and evidence indicates that dogs and humans may potentially serve as a source of infection to sand fly vectors and accordingly dogs can be recognized as a probable animal reservoir for the Leishmania infection in the endemic region in Bangladesh. However, further studies are required to determine the ability of dogs to transmit the L. donovani to the vector sand fly in nature and its evidence on ‘One Health’ perspectives. Keywords: Visceral leishmaniasis, Endemic region, Stray dogs, rK39 strip test, Giemsa’s stained liver and spleen impression smears, PCR, Reservoir host, Mymensingh

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Role of Circulating Immune Complexes in the Pathogenesis of Canine Leishmaniasis: New Players in Vaccine Development

Microorganisms ◽

10.3390/microorganisms9040712 ◽

2021 ◽

Vol 9 (4) ◽

pp. 712

Author(s):

Cristina Cacheiro-Llaguno ◽

Nuria Parody ◽

Marta R. Escutia ◽

Jerónimo Carnés

Keyword(s):

Immune Complexes ◽

Vaccine Development ◽

Correct Diagnosis ◽

Circulating Immune Complexes ◽

Response To Treatment ◽

Canine Leishmaniasis ◽

Leishmania Infection ◽

Serum Samples ◽

Humoral Immune

During canine visceral leishmaniasis (CanL), due to Leishmania infantum (L. infantum), uncontrolled infection leads to a strong humoral immune response. As a consequence of the production of high antibody levels and the prolonged presence of parasite antigens, circulating immune complexes (CIC) are formed, which can be deposited in certain organs and tissues, inducing vasculitis, uveitis, dermatitis and especially glomerulonephritis and renal failure. A method to detect CIC and quantify their levels in serum samples from dogs infected with L. infantum has been recently described. It allowed demonstration of a correlation between CIC levels and disease severity. Thus, CIC measurement may be useful for diagnosis, assessment of disease progression and monitoring response to treatment. This is an interesting finding, considering that there remains an urgent need for identification of novel biomarkers to achieve a correct diagnosis and for optimal disease staging of dogs suffering from Leishmania infection. The objective of the present review is to shed light on the role of CIC in CanL, as well as to highlight their potential use not only as diagnostic and prognostic biomarkers but also as a valuable tool in vaccine development and new immunotherapy strategies to prevent or control disease outcome.

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Presence of Leishmania amastigotes in peritoneal fluid of a dog with leishmaniasis from Alagoas, Northeast Brazil

Revista do Instituto de Medicina Tropical de São Paulo ◽

10.1590/s0036-46652006000400009 ◽

2006 ◽

Vol 48 (4) ◽

pp. 219-221 ◽

Cited By ~ 5

Author(s):

Filipe Dantas-Torres

Keyword(s):

Bone Marrow ◽

Lymph Node ◽

Peritoneal Fluid ◽

Skin Ulcer ◽

Popliteal Lymph Node ◽

Canine Leishmaniasis ◽

Leishmania Infection ◽

Intracellular Amastigotes ◽

Skin Ulcers ◽

Loss Of Weight

The goal of this short communication is to report the uncommon presence of intracellular amastigotes of Leishmania in peritoneal fluid of a dog with leishmaniasis from Alagoas State, Brazil. Physical examination of an adult male rottweiler suspected to be suffering of leishmaniasis revealed severe loss of weight, ascitis, splenomegaly, moderately enlarged lymph nodes, onychogryphosis, generalized alopecia, skin ulcers on the posterior limbs, and conjunctivitis. Samples of bone marrow, popliteal lymph node, skin ulcer, and peritoneal fluid were collected and smears of each sample were prepared and stained with hematoxylin and eosin. Numerous amastigotes were detected in bone marrow, popliteal lymph node, and skin ulcer smears. Smears of peritoneal fluid revealed the unusual presence of several free and intracellular amastigotes of Leishmania. Future studies are needed to determine whether the cytology of ascitic fluid represents a useful tool for diagnosis Leishmania infection in ascitic dogs, particularly in those living in areas where canine leishmaniasis is enzootic.

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Detection of Leishmania tarentolae in lizards, sand flies and dogs in southern Italy, where Leishmania infantum is endemic: hindrances and opportunities

Parasites & Vectors ◽

10.1186/s13071-021-04973-2 ◽

2021 ◽

Vol 14 (1) ◽

Author(s):

Jairo Alfonso Mendoza-Roldan ◽

Maria Stefania Latrofa ◽

Roberta Iatta ◽

Ranju R. S. Manoj ◽

Rossella Panarese ◽

...

Keyword(s):

Leishmania Infantum ◽

Southern Italy ◽

Antibody Test ◽

Canine Leishmaniasis ◽

Sand Flies ◽

Leishmania Tarentolae ◽

Tissue Samples ◽

Podarcis Siculus ◽

Vertebrate Hosts ◽

And Control

Abstract Background Leishmania tarentolae is a protozoan isolated from geckoes (Tarentola annularis, Tarentola mauritanica), which is considered non-pathogenic and is transmitted by herpetophilic Sergentomyia spp. sand flies. This species occurs in sympatry with Leishmania infantum in areas where canine leishmaniasis is endemic. In the present study, we investigated the circulation of L. tarentolae and L. infantum in sand flies, dogs and lizards in a dog shelter in southern Italy, where canine leishmaniasis by L. infantum is endemic. Methods Sheltered dogs (n = 100) negative for Leishmania spp. (March 2020) were screened by immunofluorescence antibody test (IFAT) using promastigotes of both species at two time points (June 2020 and March 2021). Whole blood from dogs, tissues of Podarcis siculus lizards (n = 28) and sand flies (n = 2306) were also sampled and tested by a duplex real-time PCR (dqPCR). Host blood meal was assessed in sand flies by PCR. Results Overall, 16 dogs became positive for L. infantum and/or L. tarentolae by IFAT at one or both sampling periods. One canine blood sample was positive for L. infantum, whilst two for L. tarentolae by dqPCR. At the cytology of lizard blood, Leishmania spp. amastigote-like forms were detected in erythrocytes. Twenty-two tissue samples, mostly lung (21.4%), scored molecularly positive for L. tarentolae, corresponding to 10 lizards (i.e., 35.7%). Of the female Sergentomyia minuta sampled (n = 1252), 158 scored positive for L. tarentolae, four for L. infantum, and one co-infected. Two Phlebotomus perniciosus (out of 29 females) were positive for L. tarentolae. Engorged S. minuta (n = 10) fed on humans, and one P. perniciosus, positive for L. tarentolae, on lagomorphs. Conclusions Dogs and lacertid lizards (Podarcis siculus) were herein found for the first time infected by L. tarentolae. The detection of both L. tarentolae and L. infantum in S. minuta and P. perniciosus suggests their sympatric circulation, with a potential overlap in vertebrate hosts. The interactions between L. tarentolae and L. infantum should be further investigated in both vectors and vertebrate hosts to understand the potential implications for the diagnosis and control of canine leishmaniasis in endemic areas. Graphical abstract

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Evaluation of the performance of three serological tests for diagnosis of Leishmania infantum infection in dogs using latent class analysis

Revista Brasileira de Parasitologia Veterinária ◽

10.1590/s1984-29612020105 ◽

2020 ◽

Vol 29 (4) ◽

Author(s):

Asier Basurco ◽

Alda Natale ◽

Katia Capello ◽

Antonio Fernández ◽

María Teresa Verde ◽

...

Keyword(s):

Sensitivity And Specificity ◽

Leishmania Infantum ◽

Latent Class ◽

Rapid Test ◽

Antibody Test ◽

Latent Class Models ◽

Canine Leishmaniasis ◽

Enzyme Linked Immunosorbent Assay ◽

Serum Samples ◽

Serological Tests

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.

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Comparative Study of Real-Time PCR (TaqMan Probe and Sybr Green), Serological Techniques (ELISA, IFA and DAT) and Clinical Signs Evaluation, for the Diagnosis of Canine Leishmaniasis in Experimentally Infected Dogs

Microorganisms ◽

10.3390/microorganisms9122627 ◽

2021 ◽

Vol 9 (12) ◽

pp. 2627

Author(s):

María Paz Peris ◽

Adriana Esteban-Gil ◽

Paula Ortega-Hernández ◽

Mariano Morales ◽

Nabil Halaihel ◽

...

Keyword(s):

Bone Marrow ◽

Lymph Node ◽

Real Time ◽

Correct Diagnosis ◽

Clinical Signs ◽

Sybr Green ◽

Taqman Probe ◽

Disease Stage ◽

Canine Leishmaniasis ◽

Leishmania Infection

Canine leishmaniasis (CanL) diagnosis is not fully resolved. Currently, two specific methodologies are in continuous development, the detection of the parasite DNA or RNA in target organs and the detection of specific antibodies against Leishmania sp. For a correct diagnosis, it has been shown that the joint use of this type of test is necessary. In this work, a Sybr Green and a TaqMan Probe based on real time PCRs (qPCR) was performed for the detection of Leishmania sp. in order to correlate the results with clinicopathological and serological evaluations (IFA, ELISA and DAT) to propose an optimal biological sample to be used to detect the parasite in both early and late stages of the infection. A total of four samples were processed: conjunctival swabs, popliteal lymph node aspirates, bone marrow aspirates, and peripheral blood from experimentally infected dogs belonging to a larger study. Our results indicated that a single non-invasive sample (conjunctival swab) and the application of both types of qPCR would be reliable for determining Leishmania infection as well as the disease stage in dogs, thus avoiding bone marrow, lymph node aspirate or blood samples collection.

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Field Application of Serodiagnostics To Identify Elephants with Tuberculosis prior to Case Confirmation by Culture

Clinical and Vaccine Immunology ◽

10.1128/cvi.00163-12 ◽

2012 ◽

Vol 19 (8) ◽

pp. 1269-1275 ◽

Cited By ~ 25

Author(s):

Konstantin P. Lyashchenko ◽

Rena Greenwald ◽

Javan Esfandiari ◽

Susan Mikota ◽

Michele Miller ◽

...

Keyword(s):

Diagnostic Algorithm ◽

Antibody Test ◽

Field Application ◽

Antibody Detection ◽

Test Accuracy ◽

Serum Samples ◽

Tissue Samples ◽

Content Type ◽

Treatment Information ◽

Posttreatment Period

ABSTRACTThree serologic methods for antibody detection in elephant tuberculosis (TB), the multiantigen print immunoassay (MAPIA), ElephantTB STAT-PAK kit, and DPP VetTB test, were evaluated using serial serum samples from 14 captive elephants infected withMycobacterium tuberculosisin 5 countries. In all cases, serological testing was performed prior to the diagnosis of TB by mycobacterial culture of trunk wash or tissue samples collected at necropsy. All elephants produced antibody responses toM. tuberculosisantigens, with 13/14 recognizing ESAT-6 and/or CFP10 proteins. The findings supported the high serodiagnostic test accuracy in detecting infections months to years beforeM. tuberculosiscould be isolated from elephants. The MAPIA and/or DPP VetTB assay demonstrated the potential for monitoring antimycobacterial therapy and predicting TB relapse in treated elephants when continuously used in the posttreatment period. History of exposure to TB and past treatment information should be taken into consideration for proper interpretation of the antibody test results. Data suggest that the more frequent trunk wash culture testing of seropositive elephants may enhance the efficiency of the TB diagnostic algorithm, leading to earlier treatment with improved outcomes.

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Feline Leishmania infection in a canine leishmaniasis endemic region, Portugal

Veterinary Parasitology ◽

10.1016/j.vetpar.2010.08.030 ◽

2010 ◽

Vol 174 (3-4) ◽

pp. 336-340 ◽

Cited By ~ 56

Author(s):

C. Maia ◽

J. Gomes ◽

J. Cristóvão ◽

M. Nunes ◽

A. Martins ◽

...

Keyword(s):

Canine Leishmaniasis ◽

Leishmania Infection ◽

Endemic Region

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Comparison of Porcine Alveolar Macrophages and CL 2621 for the Detection of Porcine Reproductive and Respiratory Syndrome (PRRS) Virus and Anti-PRRS Antibody

Journal of Veterinary Diagnostic Investigation ◽

10.1177/104063879300500204 ◽

1993 ◽

Vol 5 (2) ◽

pp. 163-165 ◽

Cited By ~ 71

Author(s):

Elida M. Bautista ◽

Sagar M. Goyal ◽

In J. Yoon ◽

Han S. Joo ◽

James E. Collins

Keyword(s):

Alveolar Macrophages ◽

Virus Isolation ◽

Fluorescent Antibody ◽

Antibody Test ◽

Cell Types ◽

Clinical Samples ◽

Established Cell Line ◽

Serum Samples ◽

Tissue Samples ◽

Prrs Virus

The American and European strains of porcine reproductive and respiratory syndrome (PRRS) virus were initially isolated in an established cell line (CL 2621) and porcine alveolar macrophages (PAM), respectively. Subsequent isolation of American strains of this virus in PAM has also been reported. To determine their relative sensitivity for virus isolation, both PAM and CL 2621 cells were inoculated with 98 tissue specimens and 73 serum samples from animals suspected of having PRRS. Four of the 98 tissue samples yielded virus in both cell types, whereas 7 samples were positive only in PAM and 4 samples only in CL 2621. Of the 73 serum samples tested, 18 were positive in PAM of which only 2 were positive in CL 2621. Additionally, 82 isolates obtained initially in CL 2621 were inoculated in PAM cells, and 18 strains isolated originally in PAM were inoculated in CL 2621. Of the 82 CL 2621 isolates, 25 could not be propagated on PAM. Of the 57 that replicated in PAM, as detected by a positive test on indirect fluorescent antibody test, only 28 produced cytopathic effects and 29 did not. Of the 18 PAM isolates, 5 did not grow on CL 2621. Although PAM were relatively more sensitive for virus isolation, their failure to support the growth of certain strains of PRRS virus indicates the existence of variants among PRRS virus strains, and both PAM and CL 2621 should be used for virus isolation from clinical samples. In addition, the sensitivity of these 2 cell types was compared for the detection of fluorescent antibodies to PRRS virus using 179 serum samples from PRRS-infected animals. The results were comparable in both cell systems.

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Platelet-derived growth factor beta is a potent inflammatory driver in paediatric high-grade glioma

Brain ◽

10.1093/brain/awaa382 ◽

2020 ◽

Author(s):

James L Ross ◽

Zhihong Chen ◽

Cameron J Herting ◽

Yura Grabovska ◽

Frank Szulzewsky ◽

...

Keyword(s):

Bone Marrow ◽

Median Survival ◽

High Grade Glioma ◽

Cell Types ◽

Tumour Microenvironment ◽

Pcr Analysis ◽

High Grade ◽

Tissue Samples ◽

Inflammatory Microenvironment

Abstract Paediatric high-grade gliomas (HGGs) account for the most brain tumour-related deaths in children and have a median survival of 12–15 months. One promising avenue of research is the development of novel therapies targeting the properties of non-neoplastic cell-types within the tumour such as tumour associated macrophages (TAMs). TAMs are immunosuppressive and promote tumour malignancy in adult HGG; however, in paediatric medulloblastoma, TAMs exhibit anti-tumour properties. Much is known about TAMs in adult HGG, yet little is known about them in the paediatric setting. This raises the question of whether paediatric HGGs possess a distinct constituency of TAMs because of their unique genetic landscapes. Using human paediatric HGG tissue samples and murine models of paediatric HGG, we demonstrate diffuse midline gliomas possess a greater inflammatory gene expression profile compared to hemispheric paediatric HGGs. We also show despite possessing sparse T-cell infiltration, human paediatric HGGs possess high infiltration of IBA1+ TAMs. CD31, PDGFRβ, and PDGFB all strongly correlate with IBA1+ TAM infiltration. To investigate the TAM population, we used the RCAS/tv-a system to recapitulate paediatric HGG in newborn immunocompetent mice. Tumours are induced in Nestin-positive brain cells by PDGFA or PDGFB overexpression with Cdkn2a or Tp53 co-mutations. Tumours driven by PDGFB have a significantly lower median survival compared to PDGFA-driven tumours and have increased TAM infiltration. NanoString and quantitative PCR analysis indicates PDGFB-driven tumours have a highly inflammatory microenvironment characterized by high chemokine expression. In vitro bone marrow-derived monocyte and microglial cultures demonstrate bone marrow-derived monocytes are most responsible for the production of inflammatory signals in the tumour microenvironment in response to PDGFB stimulation. Lastly, using knockout mice deficient for individual chemokines, we demonstrate the feasibility of reducing TAM infiltration and prolonging survival in both PDGFA and PDGFB-driven tumours. We identify CCL3 as a potential key chemokine in these processes in both humans and mice. Together, these studies provide evidence for the potent inflammatory effects PDGFB has in paediatric HGGs.

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