Evaluation of Cytopathological Techniques for the Diagnosis of Canine Visceral Leishmaniosis with Lymph Node Samples

This study aimed to evaluate the efficacy of conventional smear cytology (SC), liquid-based cytology (LBC), cell block (CB) stained with haematoxylin and eosin (HE) and immunocytochemistry (ICC), and formalin-fixed paraffin wax-embedded tissue immunohistochemistry (FFPE–IHC) compared with serology and polymerase chain reaction for the diagnosis of canine visceral leishmaniosis (CVL) in lymphoid tissue. The use of a preservative medium and centrifugation for cytological samples reduced the number of unsatisfactory artefacts/background. Moreover, LBC allowed excellent cellular preservation and the application of ancillary techniques, such as CB and ICC. SC was the most accurate morphological diagnostic method (45.0%). CB–ICC alone or associated with SC demonstrated significantly higher sensitivity (70.0% and 72.0%, respectively) when compared with SC alone (34.00%). CB–ICC was found to be more effective in the detection of infected animals with mild clinical signs, similar to FFPE–IHC. The specificity and positive predictive value were similar between all methods. Finally, the detection limit for CB–ICC and SC + CB–ICC was identical (18.46 amastigotes/mm2). Our study suggests that CB–ICC is a promising tool for improvement of the cytopathological diagnosis of CVL and may be applied in routine epidemiological screening.
Source: Journal of Comparative Pathology - Category: Pathology Source Type: research

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Authors: Çulha G, Kaya T, Gülbol Duran G, Urhan Küçük M, Doğramacı AÇ, Tiyekli Çelik D Abstract Leishmaniasis is a parasitic disease that is transmitted to humans by the bites of infected female phlebotomine sandflies. In the diagnosis of cutaneous leishmaniasis (CL), in the smear samples, the demonstration of the parasite by microscope remains a gold standard method. However, it becomes difficult to diagnose the parasite since the number of amastigotes in chronic cases with a lesion of one year or longer is very low. Due to many factor such as patients primarily do not ...
Source: Mikrobiyoloji Bulteni - Category: Microbiology Tags: Mikrobiyol Bul Source Type: research
This study aimed to investigate the circulation of L. infantum among sylvatic hosts in Sicily (southern Italy), a highly endemic area for canine leishmaniosis, through serological and molecular tools. Target tissues (skin, spleen, lymph nodes) collected from 71 European rabbits, 2 European hares, 7 red foxes, 11 European wildcats and 1 pin. marten, were qPCR analysed for the detection of L. infantum DNA. Additionally, 40 rabbits, older than one year, were serologically screened for specific anti-Leishmania antibodies. Leishmania infantum was molecularly diagnosed in 5.4% (n = 5) of the examined animals...
Source: Comparative Immunology, Microbiology and Infectious Diseases - Category: Infectious Diseases Source Type: research
This study aimed to investigate the circulation of L. infantum among sylvatic hosts in Sicily (southern Italy), a highly endemic area for canine leishmaniosis, through serological and molecular tools. Target tissues (skin, spleen, lymph nodes) collected from 71 European rabbits, 2 European hares, 7 red foxes, 11 European wildcats and 1 pine marten, were qPCR analysed for the detection of L. infantum DNA. Additionally, 40 rabbits, older than one year, were serologically screened for specific anti-Leishmania antibodies. Leishmania infantum was molecularly diagnosed in 5.4% (n = 5) of the examined animals (3/71 European r...
Source: Comparative immunology, microbiology and infectious diseases. - Category: Allergy & Immunology Authors: Tags: Comp Immunol Microbiol Infect Dis Source Type: research
Conclusions Nucleoside hydrolases are vital enzymes for the replication of Leishmania, conserved phylogenetic marker of the genus and strong-specific immunogens. We demonstrated that NH36 is an excellent target for chemotherapy of visceral leishmaniasis. Searching for the most immunogenic fraction of promastigotes of Leishmania we described the FML antigen of L. (L.) donovani, that has as its main component, the NH36 Nucleoside hydrolase. We developed second–generation vaccines with the FML and the NH36 native antigens, and with the NH36 recombinant protein. In addition, we obtained a third generation vaccine based ...
Source: Frontiers in Immunology - Category: Allergy & Immunology Source Type: research
il JF Abstract Cutaneous leishmaniasis is endemic in Salta province, which belongs to the northwest of Argentina. Leishmania spp. DNA from Giemsa-stained slides of up to 12 years in storage of patients from Salta was characterized through PCR-restriction fragment length polymorphism. One hundred smears positive for microscopy, classified in a semiquantitative scale for amastigote density, were analyzed. Also, Leishmanin skin test (LST) results were included. DNA extraction was carried out applying lysis buffer with proteinase K, and then DNA was amplified with ribosomal internal transcribed spacer 1 primers. PCR p...
Source: The American Journal of Tropical Medicine and Hygiene - Category: Tropical Medicine Authors: Tags: Am J Trop Med Hyg Source Type: research
In this study, the importance of support for the diagnosis of different microbiological methods used in the diagnosis of leishmaniasis infection which occurred during the outbreaks of the disease has been put forward. In addition, it was aimed to draw attention to the importance of imported CL cases in our country diagnosed in five truck drivers making transportation from Hatay to Turkmenistan, Syria, Saudi Arabia, Iran, Georgia, Uzbekistan and Azerbaijan. PMID: 30156518 [PubMed - in process]
Source: Mikrobiyoloji Bulteni - Category: Microbiology Tags: Mikrobiyol Bul Source Type: research
Background: Visceral (VL) and cutaneous (CL) leishmaniasis are seen in Turkey endemically and CL caused by Leishmania tropica is an important public health problem in southeastern as well as other regions of Turkey. The diagnosis has been usually made by clinical view of lesion and/or parasitological using lesion aspiration smears. Histological examination does not, however, always reveal the parasite in the skin biopsy, particularly in chronic lesions. Besides this, due to CL infections caused by different species in endemic areas, diagnostic methods enabling species identification are in great need.
Source: International Journal of Infectious Diseases - Category: Infectious Diseases Authors: Tags: UMP. 614 Source Type: research
This study was aimed to investigate the parasite species and genetic diversity of isolates obtained from CL patients based on the N-acetylglucosamine-1-phosphate transferase (nagt) gene. Methods: Exudate materials were collected from the swollen margin of the skin lesions of the patients suspected with CL who were referred to health centers laboratory of Mehran, Dehloran, Ilam and Malekshahi cities in the Ilam province. Demographic data were collected through a questionnaire. Smears were stained and examined microscopically. In total, 62 parasitologically positive samples were subjected to PCR-RFLP of nagt gene for ide...
Source: Journal of Vector Borne Diseases - Category: Infectious Diseases Tags: J Vector Borne Dis Source Type: research
Abstract The current four year study was undertaken to investigate the clinical and epidemiological features of Leishmania (Viannia) guyanensis infections in Valle Hermoso, Santo Domingo de Los Tsachilas province, north-central Pacific areas of Ecuador. A total of 155 parasitologically confirmed (Leishmania-amastigote-positive) clinical cases diagnosed at a rural health center during January 2014-December 2017 were analyzed thoroughly. Molecular characterization of the causative Leishmania parasites from different endemic sites within the study areas was performed by PCR amplification of cytochrome b (cyt b) seque...
Source: Acta Tropica - Category: Infectious Diseases Authors: Tags: Acta Trop Source Type: research
Authors: Riboldi E, Carvalho F, Romão PRT, Barcellos RB, Bello GL, Ramos RR, de Oliveira RT, Júnior JPA, Rossetti ML, Dallegrave E Abstract In Brazil, visceral leishmaniasis (VL) is expanding and becoming urbanized, especially in non-endemic areas such as the State of Rio Grande do Sul. Considering that infected dogs are the main reservoir for zoonotic VL, this study evaluated the prevalence of canine visceral leishmaniasis (CVL) in the metropolitan area of Porto Alegre, a new area of expansion of VL in Brazil. Serum and plasma from 405 asymptomatic dogs from the municipalities of Canoas (n=107), S&at...
Source: Korean Journal of Parasitology - Category: Parasitology Tags: Korean J Parasitol Source Type: research
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