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The main objectives of our study were to highlight the annual incidence of Canine Leishmaniasis (CanL) in Algeria. A retrospective study was conducted on CanL in Algeria between 2010–2019, by means of a questionnaire survey of 30 canine veterinarians practising in the North central region, Algiers. The retrospective study showed 78 cases of Canine Leishmaniasis (2010 and 2019), with an average of 13.73 per year. The highest mean was observed in 2013:3.400, [1.517–5.283] 95%, with a significant difference (p = 0.028). The cross-sectional survey showed that 44.4% of canine leishmaniasis cases are prevalent in spring and summer. Weight loss was the most frequent clinical sign of 88.9% and skin sampling was the most used diagnostic test to confirm this disease 66.9%. Our study provides a general view of the situation of this disease, particularly the number of cases, and their geographical distribution in Algeria. Canine leishmaniasis (CanL) is a serious zoonosis. Strict measures must be put in place. [ABSTRACT FROM AUTHOR]

Canine leishmaniasis (CanL) is a zoonotic disease caused by infection with Leishmania infantum and transmitted by female phlebotomine sandflies. It is prevalent in the Mediterranean basin, including the Maghreb region which comprises of Algeria, Tunisia, and Morocco. The environmental conditions in this area provide a suitable habitat for the proliferation of the phlebotomine sandfly, making it an endemic region for CanL. In this context, we carried out a systematic review and meta-analysis on the prevalence of canine leishmaniasis in these three Maghreb countries, identifying the risk factors for L. infantum infection. Data were collected from 27 papers published between 1973 and 2022 selected from five databases: PubMed, Google Scholar, ResearchGate, ScienceDirect and Scopus. Our selection criteria included both descriptive and analytical studies on canine leishmaniasis in Maghreb, as well as epidemiological studies that reported the prevalence of leishmaniasis infection. The meta-analysis was performed using Review Manager (RevMan) software version 5.4.1. based on predefined inclusion criteria. A total of 11,736 dogs were included in the analysis, among which 2481 were tested Leishmania positive cases. Over the years, the highest prevalence (68.32%) was recorded in Tiaret (Algeria) and the lowest (4.3%) in Tunisia. The examination of the risk factors revealed a heightened incidence of the disease in dogs of shepherd breeds, living in rural environments, displaying no apparent symptoms, and falling within the age range of 2 to 4 years. The findings presented in this study contribute a significant value to the exploration of leishmaniasis in this region, particularly by shedding light on the associated risk factors. Ultimately, substantial endeavors are warranted to enhance prevention, treatment, and overall control of the disease.

Human zoonotic visceral leishmaniasis (ZVL) and canine leishmaniasis (CanL) constitute a major public and veterinary health concern and are both caused by the infection with the protozoan parasite Leishmania infantum. One of the main target organs in CanL is the liver. This complex organ, composed of various highly specialized cell types, has garnered significant attention from the scientific community as a crucial player in innate immune functions. In the context of CanL, liver infection by parasites and the host immune response generated strongly influence the disease outcome. Thus, taking advantage of a co-culture system involving canine hepatocytes and L. infantum-infected autologous Kupffer cells (KCs), allowing cell-to-cell interaction, the current report aims to shed light on the hepatocyte-KCs immune interaction. The co-culture of infected KCs with hepatocytes revealed a vital role of these cells in the activation of a local immune response against L. infantum parasites. Although KCs alone can be immunologically silenced by L. infantum infection, the cell-to-cell interaction with hepatocytes in co-culture can lead to local immune activation. In co-culture it was observed gene expression increased the number of innate immune receptors, specifically cell membrane TLR2 and cytoplasmatic NOD1 along with high TNF-α generation. Altogether, these results suggest that the immune response generated in co-culture could induce the recruitment of other circulating cells to contain and contribute to the resolution of the infection in the liver. This work also enhances our understanding of the liver as a vital organ in innate immunity within the context of CanL. [ABSTRACT FROM AUTHOR]

Simple Summary: This study evaluates the potential adverse effects associated with meglumine antimoniate (aNm) in the treatment of canine leishmaniasis through both a retrospective analysis and a prospective study. The retrospective study comprised records of 87 dogs treated with aNm. Adverse effects during treatment emerged in about one third of dogs, including local reactions at the injection site, injection site pain-related systemic reaction, systemic disease due to renal function worsening, acute pancreatitis, gastrointestinal self-limiting signs or severe idiosyncratic skin reactions. Half of these animals required treatment suspension. The prospective study included 16 dogs (LeishVet stages II and III) treated with the same aNm; of these, 2 dogs were excluded for severe reactions at the injection site and 4 dogs reported mild and transient adverse events. In the prospective study, treatment efficacy was also evaluated at 1-year follow-up. No animals showed clinical laboratory relapse during the study and interestingly, PCR turned negative between D0 and D60 in 78.5% of animals. Adverse severe events associated with aNm are possible, but the high rate of parasitic clearance supports its use as first line treatment, at least in dogs with normal renal function. Antimoniate therapy, in association with allopurinol, is one of the first-line treatments of canine leishmaniasis (CanL). This study evaluates the potential adverse effects associated with aNm in the treatment of CanL through both a retrospective analysis and a long-term prospective study also aimed to investigate its efficacy. The retrospective study reviewed records of 87 dogs with CanL with at least one follow-up available during or at the end of therapy with aNm (Glucantime®) at a dose of 50 mg/kg administered subcutaneously twice a day in association with allopurinol. In total, 29.8% of dogs showed adverse effects during treatment as local reactions at the injection site (n = 6), severe systemic reaction to pain (originating from the inoculation site) with depression and anorexia (n = 4), systemic disease due to renal function worsening (n = 4), acute pancreatitis (n = 1), diarrhea (n = 5), vomiting (n = 3) and severe idiosyncratic skin reactions (n = 3). Of these dogs, 13 (14.9%) required treatment suspension. The prospective study included 16 dogs, selected among the LeishVet stages II and III CKD IRIS stage 1 (International Renal Interest Society staging of canine Chronic Kidney Disease) and treated with the same aNm plus allopurinol protocol as in the retrospective study and observed for 360 days; 2 dogs were excluded for severe reactions at the injection site. Mild and transient adverse events were reported in the other 4 dogs. The criteria used to evaluate the efficacy of treatment with aNm were as follows: a reduction in the clinical score and improvement and/or normalization of laboratory parameters, negativization of PCR on the bone marrow samples and disease-free interval time. The proportion of reduction in the clinical score reached 91.9% at D180. No animals showed clinical laboratory relapse during the whole study duration and interestingly, the PCR results showed complete negativity between D0 and D60 in 78.5% of animals. Veterinarians must be vigilant regarding the potentially serious adverse effects associated with aNm and promptly stop drug administration if unexpected clinical manifestations occur. On the other hand, they should not discard its use for CanL treatment since it is confirmed that aNm in association with allopurinol is highly effective in controlling CanL. [ABSTRACT FROM AUTHOR]

Simple Summary: Leishmania infantum is a single-celled trypanosomatid responsible for canine leishmaniasis, a serious disease that causes a generalized inflammatory reaction including, among others, lymphadenopathy, hepatomegaly, and splenomegaly in dogs. Increasing information on leishmaniasis in wildlife shows that many mammalian species can act as reservoirs, being a serious threat to domestic dog populations and public health. Our case describes a female wolf suffering from lameness due to an ulcerative wound on the right forepaw, with hyper-gammaglobulinemia, high serum GPT levels, and positive ELISA tests for antibodies to Leishmania infantum antigens as confirmed by PCR. The wolf responded positively to conventional treatment of leishmaniasis, and all altered parameters were restored. Leishmaniasis in wild canids is a vector-borne disease caused in Europe by the protozoan parasite Leishmania infantum. To date, there is limited information on clinical signs and laboratory abnormalities in wolves due to leishmaniasis. The current clinical case report described a female Iberian wolf (Canis lupus signatus) housed in semi-captivity conditions at the Centro del Lobo Ibérico "Félix Rodríguez de la Fuente", in Robledo de Sanabria, Zamora (Spain), with an interdigital ulcerous wound at the right forepaw, hyper-gammaglobulinemia, and abnormal liver blood parameters. Definitive serodiagnosis of leishmaniasis was established using antileishmanial serum antibodies and PCR analysis of different biological samples. A gold-standard anti-L. infantum treatment protocol consisting in subcutaneous meglumine antimoniate and oral allopurinol combination was installed. However, the presence of pain at the site of injection due to meglumine antimoniate administration forced its substitution by oral miltefosine. A progressive reduction of the levels of anti-L. infantum serum antibodies and the concentrations of gamma-globulin fraction was detected after antileishmanial treatment as well as a decline of liver GPT. To our knowledge, this is the first case of leishmaniasis diagnosed in a wolf housed in semi-captivity conditions, with the condition subsequently treated and successfully cured. [ABSTRACT FROM AUTHOR]

The diagnosis of canine leishmaniasis (CanL) still represents a challenge due to the variable clinical manifestations and the large number of asymptomatic dogs. Serological tests are most commonly used to detect infected animals, revealing anti-Leishmania antibodies, mainly of the IgG isotype. Recently, a new diagnostic antigen, rKLi8.3, containing 8.3 kinesin tandem repeats (TR) from a Leishmania infantum strain from Sudan, has been shown to provide excellent specificity and sensitivity for the detection of Leishmania-infected humans and dogs. However, asymptomatic animals with very low antibody titers are often difficult to detect by serodiagnosis. Thus, we wondered whether the addition of an anti-IgG-enhancing step in the protein A/G-based rKLi8.3-ELISA will improve the diagnostic performance without decreasing the specificity. For this, parasitologically confirmed CanL cases with low or high clinical scores, uninfected healthy controls and dogs with other infections were tested by rKLi8.3-ELISA as well as two different immunochromatographic rapid tests, rKLi8.3-lateral flow test (LFT) and Dual Path Platform (DPP®) based on the rK28 antigen. Our results show that the diagnostic accuracies of the rKLi8.3-ELISA and LFT were similar to that of DPP, missing several asymptomatic animals. However, the addition of a secondary, amplifying anti-dog IgG antibody in the protein A/G-based rKLi8.3-ELISA enabled the detection of nearly all asymptomatic dogs without compromising its specificity. [ABSTRACT FROM AUTHOR]

Human zoonotic visceral leishmaniasis (ZVL) and canine leishmaniasis (CanL) constitute a major public and veterinary health concern and are both caused by the infection with the protozoan parasite Leishmania infantum. One of the main target organs in CanL is the liver. This complex organ, composed of various highly specialized cell types, has garnered significant attention from the scientific community as a crucial player in innate immune functions. In the context of CanL, liver infection by parasites and the host immune response generated strongly influence the disease outcome. Thus, taking advantage of a co-culture system involving canine hepatocytes and L. infantum-infected autologous Kupffer cells (KCs), allowing cell-to-cell interaction, the current report aims to shed light on the hepatocyte-KCs immune interaction. The co-culture of infected KCs with hepatocytes revealed a vital role of these cells in the activation of a local immune response against L. infantum parasites. Although KCs alone can be immunologically silenced by L. infantum infection, the cell-to-cell interaction with hepatocytes in co-culture can lead to local immune activation. In co-culture it was observed gene expression increased the number of innate immune receptors, specifically cell membrane TLR2 and cytoplasmatic NOD1 along with high TNF-α generation. Altogether, these results suggest that the immune response generated in co-culture could induce the recruitment of other circulating cells to contain and contribute to the resolution of the infection in the liver. This work also enhances our understanding of the liver as a vital organ in innate immunity within the context of CanL.

Antimoniate therapy, in association with allopurinol, is one of the first-line treatments of canine leishmaniasis (CanL). This study evaluates the potential adverse effects associated with aNm in the treatment of CanL through both a retrospective analysis and a long-term prospective study also aimed to investigate its efficacy. The retrospective study reviewed records of 87 dogs with CanL with at least one follow-up available during or at the end of therapy with aNm (Glucantime®) at a dose of 50 mg/kg administered subcutaneously twice a day in association with allopurinol. In total, 29.8% of dogs showed adverse effects during treatment as local reactions at the injection site (n = 6), severe systemic reaction to pain (originating from the inoculation site) with depression and anorexia (n = 4), systemic disease due to renal function worsening (n = 4), acute pancreatitis (n = 1), diarrhea (n = 5), vomiting (n = 3) and severe idiosyncratic skin reactions (n = 3). Of these dogs, 13 (14.9%) required treatment suspension. The prospective study included 16 dogs, selected among the LeishVet stages II and III CKD IRIS stage 1 (International Renal Interest Society staging of canine Chronic Kidney Disease) and treated with the same aNm plus allopurinol protocol as in the retrospective study and observed for 360 days; 2 dogs were excluded for severe reactions at the injection site. Mild and transient adverse events were reported in the other 4 dogs. The criteria used to evaluate the efficacy of treatment with aNm were as follows: a reduction in the clinical score and improvement and/or normalization of laboratory parameters, negativization of PCR on the bone marrow samples and disease-free interval time. The proportion of reduction in the clinical score reached 91.9% at D180. No animals showed clinical laboratory relapse during the whole study duration and interestingly, the PCR results showed complete negativity between D0 and D60 in 78.5% of animals. Veterinarians must be vigilant regarding the potentially serious adverse effects associated with aNm and promptly stop drug administration if unexpected clinical manifestations occur. On the other hand, they should not discard its use for CanL treatment since it is confirmed that aNm in association with allopurinol is highly effective in controlling CanL.

ABSTRACT: Canine leishmaniasis (CanL) is a multifaceted disease triggered by the protozoan Leishmania infantum, characterized by diverse clinical presentations, including osteoarticular complications. Immune-mediated joint diseases invariably initiate at the synovial membrane, implicating its pivotal role in arthritis pathogenesis. This study aimed to investigate the influence of natural L. infantum infection on synovial fluid characteristics and the expression of immune markers, including TLR-2, FOXP3, and COX-2, in the synovial membrane. Twenty naturally infected dogs (NID) with L. infantum were sourced from the Zoonosis Surveillance Unit (ZSU). Clinical-orthopedic assessments were conducted, encompassing lameness, joint edema, crepitus, patellar luxation, and the drawer test. Synovial fluid (SF) parameters, including volume, appearance, viscosity, total nucleated cell count (TNC), neutrophil count, and total protein (TP) content, were determined. After anesthesia and euthanasia, synovial membrane specimens were obtained. SF protein concentrations categorized dogs into three groups: GI (2 to 2.5g/dL), GII (2.5 to 6.0g/dL), and GIII (>6g/dL). Inflammatory infiltrates and synovial membrane changes were assessed, and immunohistochemistry evaluated TLR-2, FOXP3, and COX-2 marker expressions. Clinical evaluations revealed various osteoarticular abnormalities in NID dogs, including lameness (55%), joint edema (25%), crepitus (30%), patellar luxation (20%), and positive drawer test (25%). Post mortem examinations revealed bilateral subchondral bone, meniscus, and trochlea erosion in 30% of cases. Amastigotes of L. infantum were identified extracellularly and within macrophages (60%). An inflammatory infiltrate was predominant in 70% of dogs, with varying intensity among the groups. Mononuclear cells, chiefly macrophages and lymphocytes, and neutrophils comprised the infiltrate. TLR-2 and COX-2 expression levels were elevated in GIII compared to GII and GI. Conversely, FOXP3 showed moderate expression in GI and minimal expression in GII and GIII. This study underscores the contributory role of L. infantum infection in the development of joint lesions in CanL. Additionally, alterations in the expression of immune markers TLR2, FOXP3, and COX2 within the synovial membrane imply the perpetuation and exacerbation of the inflammatory processes, shedding light on the intricate pathogenesis of CanL-induced arthritis.

Background: Canin leishmaniasis (CanL), mostly caused by Leishmania infantum, is one of the most important vector-borne diseases in dogs in the Mediterranean region. In this study, we aimed to determine the disease profile in this region by firstly making microscopic and then molecular analyzes in the samples taken from the dogs. Methods: Overall, 112 whole blood samples taken from dogs for clinical applications by a veterinarian in Cankırı between December 2021 and November 2022 were used. After blood collection, both thin and thick drop blood smear preparations were prepared and evaluated for Giemsa staining. L. infantum was investigated by Real time-PCR (RT-PCR) method from all blood samples. Sequence analysis and phylogenetic tree study were performed on positive samples. Results: Both microscopic and RT-PCR analyzes were performed. In both studies, 3 of the 112 samples were positive. Because of the sequence analysis, they were L. infantum. Sequence analysis was performed from the samples found 3 positive. The phylogenetic tree was drawn by making NCBI (National Center for Biotechnology) data entries of the positive samples (Accession numbers: OQ184728, OQ184729, OQ184730). Conclusion: Dogs are important, as they are reservoir of this disease. In this study, 3 (2.7%) positive Leishmaniasis was detected in dogs in Cankırı. Ultimately, this should prompt discussion about new strategies going forward to combat infection caused by Leishmania. [ABSTRACT FROM AUTHOR]

Leishmania infantum is the etiological agent of zoonotic visceral leishmaniasis (ZVL). The disease is endemic in Central and South America, Central and South East Asia, and the Mediterranean basin. Dogs are the main reservoir, with an estimated prevalence of approximately 2.5 million dogs in Southern Europe. Current treatments cause side effects, disease recurrence, and drug resistance. Therefore, the development of vaccines against canine leishmaniasis is necessary. We have generated a DNA vaccine based on the non-replicative antibiotic resistance marker-free plasmid vector pPAL that contains the encoding gene for the L. infantum activated protein kinase C receptor analog (LACK). Homologous pPALLACK prime-boost intranasal administration confers efficacious protection in Beagle dogs with a reduction of clinical signs and a statistically significant reduction of the parasite burden in the bone marrow of more than 90% of dogs after experimental infection with highly infective promastigotes. This DNA vaccine elicits a robust cellular immune response skewed towards the Th1 profile. [ABSTRACT FROM AUTHOR]

Leishmaniasis in wild canids is a vector-borne disease caused in Europe by the protozoan parasite Leishmania infantum. To date, there is limited information on clinical signs and laboratory abnormalities in wolves due to leishmaniasis. The current clinical case report described a female Iberian wolf (Canis lupus signatus) housed in semi-captivity conditions at the Centro del Lobo Ibérico “Félix Rodríguez de la Fuente”, in Robledo de Sanabria, Zamora (Spain), with an interdigital ulcerous wound at the right forepaw, hyper-gammaglobulinemia, and abnormal liver blood parameters. Definitive serodiagnosis of leishmaniasis was established using antileishmanial serum antibodies and PCR analysis of different biological samples. A gold-standard anti-L. infantum treatment protocol consisting in subcutaneous meglumine antimoniate and oral allopurinol combination was installed. However, the presence of pain at the site of injection due to meglumine antimoniate administration forced its substitution by oral miltefosine. A progressive reduction of the levels of anti-L. infantum serum antibodies and the concentrations of gamma-globulin fraction was detected after antileishmanial treatment as well as a decline of liver GPT. To our knowledge, this is the first case of leishmaniasis diagnosed in a wolf housed in semi-captivity conditions, with the condition subsequently treated and successfully cured.

Background: Canin leishmaniasis (CanL), mostly caused by Leishmania infantum, is one of the most important vector-borne diseases in dogs in the Mediterranean region. In this study, we aimed to determine the disease profile in this region by firstly making microscopic and then molecular analyzes in the samples taken from the dogs. Methods: Overall, 112 whole blood samples taken from dogs for clinical applications by a veterinarian in Cankırı between December 2021 and November 2022 were used. After blood collection, both thin and thick drop blood smear preparations were prepared and evaluated for Giemsa staining. L. infantum was investigated by Real time-PCR (RT-PCR) method from all blood samples. Sequence analysis and phylogenetic tree study were performed on positive samples. Results: Both microscopic and RT-PCR analyzes were performed. In both studies, 3 of the 112 samples were positive. Because of the sequence analysis, they were L. infantum. Sequence analysis was performed from the samples found 3 positive. The phylogenetic tree was drawn by making NCBI (National Center for Biotechnology) data entries of the positive samples (Accession numbers: OQ184728, OQ184729, OQ184730). Conclusion: Dogs are important, as they are reservoir of this disease. In this study, 3 (2.7%) positive Leishmaniasis was detected in dogs in Cankırı. Ultimately, this should prompt discussion about new strategies going forward to combat infection caused by Leishmania.

While many Bayesian state‐space models for infectious disease processes focus on population infection dynamics (eg, compartmental models), in this work we examine the evolution of infection processes and the complexities of the immune responses within the host using these techniques. We present a joint Bayesian state‐space model to better understand how the immune system contributes to the control of Leishmania infantum infections over the disease course. We use longitudinal molecular diagnostic and clinical data of a cohort of dogs to describe population progression rates and present evidence for important drivers of clinical disease. Among these results, we find evidence for the importance of co‐infection in disease progression. We also show that as dogs progress through the infection, parasite load is influenced by their age, ectoparasiticide treatment status, and serology. Furthermore, we present evidence that pathogen load information from an earlier point in time influences its future value and that the size of this effect varies depending on the clinical stage of the dog. In addition to characterizing the processes driving disease progression, we predict individual and aggregate patterns of Canine Leishmaniasis progression. Both our findings and the application to individual‐level predictions are of direct clinical relevance, presenting possible opportunities for application in veterinary practice and motivating lines of additional investigation to better understand and predict disease progression. Finally, as an important zoonotic human pathogen, these results may support future efforts to prevent and treat human Leishmaniosis. [ABSTRACT FROM AUTHOR]

Canine leishmaniasis is an important vector-borne protozoan disease in dogs that is responsible for serious deterioration in their health. In the Iberian Peninsula, as in most countries surrounding the Mediterranean Sea, canine leishmaniasis is caused by Leishmania infantum (zymodeme MON-1), a digenetic trypanosomatid that harbors in the parasitophorous vacuoles of host macrophages, causing severe lesions that can lead to death if the animals do not receive adequate treatment. Canine leishmaniasis is highly prevalent in Spain, especially in the Mediterranean coastal regions (Levante, Andalusia and the Balearic Islands), where the population of domestic dogs is very high. However, the presence of this disease has been spreading to other rural and sparsely populated latitudes, and cases of leishmaniasis have been reported for years in wildlife in northwestern Spain. This work describes for the first time the presence of wolves that tested positive for leishmaniasis in the vicinity of the Sierra de la Culebra (Zamora province, northwestern Spain), a protected sanctuary of this canid species, using PCR amplification of L. infantum DNA from different non-invasive samples such as buccal mucosa and those from both ears and hair. In addition to live animals (21), samples from carcasses of mainly roadkill animals (18) were also included and analyzed using the same technique, obtaining a positivity rate of 18 of the 39 wolves sampled (46.1%) regardless of their origin. [ABSTRACT FROM AUTHOR]

Leishmania infantum is the etiological agent of zoonotic visceral leishmaniasis (ZVL). The disease is endemic in Central and South America, Central and South East Asia, and the Mediterranean basin. Dogs are the main reservoir, with an estimated prevalence of approximately 2.5 million dogs in Southern Europe. Current treatments cause side effects, disease recurrence, and drug resistance. Therefore, the development of vaccines against canine leishmaniasis is necessary. We have generated a DNA vaccine based on the non-replicative antibiotic resistance marker-free plasmid vector pPAL that contains the encoding gene for the L. infantum activated protein kinase C receptor analog (LACK). Homologous pPAL-LACK prime-boost intranasal administration confers efficacious protection in Beagle dogs with a reduction of clinical signs and a statistically significant reduction of the parasite burden in the bone marrow of more than 90% of dogs after experimental infection with highly infective promastigotes. This DNA vaccine elicits a robust cellular immune response skewed towards the Th1 profile.

The diagnosis of canine leishmaniasis (CanL) still represents a challenge due to the variable clinical manifestations and the large number of asymptomatic dogs. Serological tests are most commonly used to detect infected animals, revealing anti-Leishmania antibodies, mainly of the IgG isotype. Recently, a new diagnostic antigen, rKLi8.3, containing 8.3 kinesin tandem repeats (TR) from a Leishmania infantum strain from Sudan, has been shown to provide excellent specificity and sensitivity for the detection of Leishmania-infected humans and dogs. However, asymptomatic animals with very low antibody titers are often difficult to detect by serodiagnosis. Thus, we wondered whether the addition of an anti-IgG-enhancing step in the protein A/G-based rKLi8.3-ELISA will improve the diagnostic performance without decreasing the specificity. For this, parasitologically confirmed CanL cases with low or high clinical scores, uninfected healthy controls and dogs with other infections were tested by rKLi8.3-ELISA as well as two different immunochromatographic rapid tests, rKLi8.3-lateral flow test (LFT) and Dual Path Platform (DPP®) based on the rK28 antigen. Our results show that the diagnostic accuracies of the rKLi8.3-ELISA and LFT were similar to that of DPP, missing several asymptomatic animals. However, the addition of a secondary, amplifying anti-dog IgG antibody in the protein A/G-based rKLi8.3-ELISA enabled the detection of nearly all asymptomatic dogs without compromising its specificity.

Dogs are considered to be the main domestic reservoir associated with the transmission of Leishmania (L.) infantum chagasi to humans in endemic areas of visceral leishmaniasis in America. However, little is known about the role of canines as a source of infection in endemic areas of nonulcerated cutaneous leishmaniasis (NUCL). Therefore, the objective of the present study was to investigate the role of dogs as a possible reservoir of the parasite in Southern Honduras. Dogs (n = 107) living with individuals affected by NUCL were clinically examined and biological material was collected for parasitological and immunological diagnosis. Most animals showed a healthy appearance and a few presented slight weight loss (64%), alopecia (7%), onychogryphosis (5%) and skin lesions (1%). The overall seroprevalence of Leishmania infection based on the DDP® quick test and/or in-house ELISA serological test was 41%. The presence of the parasite's DNA was confirmed in 94% of the dogs; however, the average parasite load in the buffy coat was low at 6.09 parasites/µL, ranging between 0.221 and 50.2. The skin of seropositive dogs examined by histopathology using paraffin sections stained by hematoxylin and immunohistochemistry did not show cutaneous lesions or parasite amastigotes. Based on the absence of parasites in the skin and the low parasite load detected in the buffy coat, it seems that the dog does not represent a good source of infection for the vector in the endemic area of NUCL transmission in Southern Honduras. Other domestic and/or wild animals should be investigated. [ABSTRACT FROM AUTHOR]

Abstract Background Leishmaniases are a group of anthropo-zoonotic parasitic diseases caused by a protozoan of the Leishmania genus, affecting both humans and other vertebrates, including dogs. L. infantum is responsible for the visceral and occasionally cutaneous form of the disease in humans and canine leishmaniasis. Previously, we have shown that L. infantum induces a mild but significant increase in endoplasmic reticulum (ER) stress expression markers to promote parasites survival in human and murine infected macrophages. Moreover, we demonstrated that the miRNA hsa-miR-346, induced by the UPR-activated transcription factor sXBP1, was significantly upregulated in human macrophages infected with different L. infantum strains. However, the ER stress response in infected dogs, which represent an important reservoir for Leishmania parasite, was described once recently, whereas the miR-346 expression was not reported before. Therefore, this study aimed to investigate these pathways in the canine macrophage-like cell line DH82 infected by Leishmania spp. and to evaluate the presence of cfa-miR-346 in plasma of non-infected and infected dogs. The DH82 cells were infected with L. infantum and L. braziliensis parasites and the expression of cfa-mir-346 and several ER stress markers was evaluated by quantitative PCR (qPCR) at different time points. Furthermore, the cfa-miR-346 was monitored in plasma collected from non-infected dogs (n = 11) and dogs naturally infected by L. infantum (n = 18). Results The results in DH82 cells showed that cfa-mir-346 was induced at both 24 h and 48 h post-infection with all Leishmania strains but not with tunicamycin, accounting for a mechanism of induction independent from sXBP1, unlike what was previously observed in human cell lines. Moreover, the cfa-miR-346 expression analysis on plasma revealed a significant increase in infected dogs compared to non-infected dogs. Conclusions Here for the first time, we report the upregulation of cfa-miR-346 induced by Leishmania infection in canine macrophage-like cells and plasma samples of naturally infected dogs. According to our results, the cfa-miR-346 appears to be linked to infection, and understanding its role and identifying its target genes could contribute to elucidate the mechanisms underlying the host–pathogen interaction in leishmaniasis.

Leishmaniasis is a significant public health concern, with dogs as the primary reservoir in urban scenarios and facilitating transmission. Diagnosing infected dogs is a crucial step for public health interventions, and the development of new diagnostic platforms can significantly enhance efforts in various regions worldwide. Given the limited availability of diagnostic methods in Colombia, this study evaluates the effectiveness of an Indirect Enzyme-Linked Immunosorbent Assay (ELISA) based on the recombinant protein rLicNTPDase-2 to detect Leishmania in infected dogs. Serum samples were collected from dogs in both endemic and non-endemic areas and classified as natural standards based on prior parasitological diagnoses. The results revealed 24 true positives (TP) and 9 true negatives (TN). Subsequently, the test was then validated with samples from symptomatic and asymptomatic animals, alongside the standards, yielding a specificity of 96 %, a sensitivity of 81 %, efficiency of 90.6 %, a positive predictive value of 92.8 %, and a negative predictive value of 89.6 %. The positive likelihood ratio (RV+) was 20, while the negative likelihood ratio (RV-) was 0.19, indicating high relevance and a robust clinical utility. The area under the curve (AUC) was 1.00, suggesting that the test has excellent discriminatory ability, significantly deviating from the reference diagonal. This is further supported by the significant difference(p < 0.0001) between TN and TP results determined by Fisher's exact test. Involving 163 animals showed 47 % positive and 46 % negative results with a significant difference (p < 0.05) in the mean optical density (OD) values between positive and negative samples. These findings indicate that the ELISA test effectively differentiates between positive and negative samples based on OD values. This study suggests that ELISA based on the recombinant antigen rLicNTPDase-2 could serve as a viable alternative for the serodiagnosis of leishmaniasis in canines in Colombia. [Display omitted] [ABSTRACT FROM AUTHOR]

Human visceral leishmaniasis has long been associated with canine leishmaniasis (CL). However, to date, there is no clear information on the status of the disease in dogs in Morocco that could be used by policymakers for the prevention of human cases. This study aims to assess the status of CL in Morocco and its risk factors through an exhaustive literature search. The meta-analysis was performed using RevMan 5.4. The main results showed that the overall prevalence of CL in Morocco is 17% (95% CI: 0.12–0.22), caused by two strains of Leishmania parasite: Leishmania tropica and L. infantum. According to the region, the maximum prevalence was reported in the coastal provinces and in the central part of the country; while, the CL risk was higher in rural area (18% [95% CI: 0.14–0.23]) and at altitude above 1000 m (23% [95% CI: − 0.08–0.53]). Regarding the intrinsic factors, the prevalence of the disease increased with the age of the dog, (30% [95% CI − 0.09–0.68) and the risk was very high in clinically asymptomatic dogs (RR = 2.08 [95% CI: 1.15–3.76]). This study is the first in Morocco indicating the CL prevalence, its geographical distribution and detailing its risk factors. These results are needed to improve management strategies for the canine reservoir of leishmaniasis in Morocco and interrupt the local transmission cycle to humans. [ABSTRACT FROM AUTHOR]

Canine leishmaniasis is a parasitic zoonosis mainly caused by L. infantum; an obligate intracellular protozoan transmitted by haematophagous insects of the genus Phlebotomus, which affects dogs and wild canids. The clinical implications of this disease are highly variable, since infected animals may remain asymptomatic (absence of observable clinical signs) or present a wide spectrum of clinical alterations and degrees of severity, including the death of the animal. Symptoms such as lymphadenomegaly, alopecia, weight loss, keratoconjunctivitis and onychogryphosis are usually the first diagnostic reference available. The objectives of this study are to evaluate the validity (sensitivity, specificity and likelihood ratios) and diagnostic utility (pre‐test probability) of the clinical signs commonly associated with canine leishmaniasis based on the prevalence in the area and to explore the combination of symptoms that best predicts the diagnosis of canine leishmaniasis. It is a matched case‐control study in the canine population of southern Spain based on the comparison of the findings collected in the clinical history and the results of the LeisSCAN quantitative ELISA. A total of 39 cases and 78 controls were analysed. Approximately 80% of the infected animals showed signs compatible with the disease. The most frequent alterations were cutaneous (64.1%), systemic (51.3%) and oculo‐nasal (30.7%). The most useful signs to support this diagnosis were alopecia and epistaxis (LR+ 6.69 and 6.0, respectively) (pre‐test leishmaniasis probability is ≥70% for prevalence ≥28% when alopecia or epistaxis is present), followed by lameness (LR+ 5.0). The combinations of signs that showed greater validity were alopecia with hyperkeratosis of the snout and alopecia with onychogryphosis (LR+ > 10). None of the observed signs or their combinations resulted useful to rule out the diagnosis (LR– 0.55 to 1.15). The results found show notable differences in the diagnostic value of the clinical signs, individually and in combination, so we believe that medical decisions should be based on their diagnostic validity (LR+) and the estimation of the pre‐test and post‐test probability. [ABSTRACT FROM AUTHOR]

Canine leishmaniasis (CanL) caused by Leishmania infantum is a prevalent vector-borne disease in the Mediterranean region, notably Algeria. This research aims to assess the epidemiology and the clinical management practices of CanL employed by veterinarians in the Skikda and Constantine regions in the northeastern part of Algeria. A retrospective survey comprising 37 open and closed questions was distributed to 139 veterinary clinics between 2022 and 2023. The objective was to collect comparable epidemiological information on clinical presentations, diagnostic approaches, treatment protocols, preventive strategies, and awareness of the disease's public health implications. The findings from this study have revealed a notable prevalence of CanL in both regions, accompanied by similar clinical manifestations. Serological techniques, particularly rapid detection kits, are predominantly utilized alongside direct methods, while euthanasia remains the primary approach to combat the disease. The results show that only a few veterinarians educate pet owners about zoonotic diseases and prevention measures (32,79 %), while most do not provide this information. This investigation sheds more light on the current status of CanL in the selected studied area and suggests the need for standardizing diagnosis and treatment of CanL using evidence-based medicine criteria to enhance disease management. This will provide a proper way of conducting a more comprehensive epidemiological assessment of the condition for improving the handling and management of CanL in Algeria., Competing Interests: Declaration of Competing Interest The authors declare that they have no known financial interests or personal relationships that could have influenced, either intentionally or unintentionally, the content and conclusions of this study. They affirm that all analyses and interpretations presented in this paper were conducted independently, without any external bias related to financial or personal interests., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Leishmaniasis is a widespread disease caused by species of the genus Leishmania. In Colombia, this zoonosis is endemic in rural areas with a high prevalence in the departments of Antioquia, Santander, Meta, Tolima and Nariño. Dogs are the most important domestic reservoirs of the pathogen, given the epidemiological importance of dogs in the control of leishmaniasis is needed to determine the prevalence of Leishmania spp. in canine population of the rural area of Ibagué and to identify potential risk factors related to the presence of this parasite. A cross-sectional study was carried out in 173 dogs from the rural area of Ibagué. Leishmania spp. was detected by amplifying the Internal Transcribed Spacer (ITS-1) and two regions of the hsp70 gene through PCR. Factor associations were calculated through the Chisquare and odds ratio. Prevalence of Leishmania spp. infection in dogs was of 91.33% (158/173), where 36.71% (58/158) of the Leishmania spp. positive dogs showed one or more clinical signs of canine leishmaniasis and 63.29% (100/158) of the dogs were asymptomatic. Factors associated with the presence of the parasite did not show significance. In addition, hsp70D-PCR was proved to be highly efficient for the detection of Leishmania spp.

IntroductionThe study of early markers to detect kidney malfunction has increased in recent years since serum markers, such as creatinine increase when there is a 75% loss of renal mass. Urinary capillary electrophoresis (UCE) is an available laboratory technique that provides an easily interpretable electrophoretic pattern. This pattern in our study has been divided into five fractions as it is done in serum: fraction 1 migrating in the albumin zone, fraction 2 in the alpha1-globulins zone, fraction 3 in the alpha2-globulins zone, fraction 4 in the beta-globulins zone, and fraction 5 in the gamma globulins zone. UCE can be useful in the early diagnosis of renal disease.Material and methodsIn this study, UCE was performed in dogs with azotemia and proteinuria due to chronic kidney disease (CKD) not related to Leishmania infantum (L. infantum) infection (G1, n = 11) and dogs with CKD related to L. infantum infection (G2, n = 17) and compared with reference intervals from healthy dogs (G0, n = 123), with the aim of comparing their phoretograms and assessing changes in the fractions of the phoretograms based on the health status of individuals.ResultsFraction 2 was statistically augmented in dogs with CKD (G1) when compared with the healthy population (G0) and dogs infected by L. infantum (G2). Fraction 3 was statistically increased in dogs with CKD (G1) and dogs infected by L. infantum (G2) compared with G0. Fraction 4 was found to be statistically decreased in dogs with CKD (G1) and dogs infected by L. infantum (G2) compared with G0. Fraction 5 was statistically higher in dogs with L. infantum (G2) compared with G0 and dogs with CKD (G1). No statistical relationship was found between the protein to creatinine ratio and different fractions from the urinary phoretogram in the study population. No statistical relationship was found between serum and urine fractions in the study population.DiscussionThe results of the present study suggest that UCE is a promising non-invasive technique that might be used as a part of the diagnostic and follow-up in dogs with kidney disease due to different pathologies.

Leishmaniases are neglected tropical diseases of humans and animals. We detected Leishmania infantum in 3 mixed-breed dogs in Zambia that had no travel history outside the country. Our findings suggest presence of and probable emergence of leishmaniasis in Zambia, indicating the need for physicians and veterinarians to consider the disease during diagnosis.

The mucosal immune system plays a pivotal role in the control of infections, as it represents the first line of defense against most pathogens, from respiratory viruses to intestinal parasites. Mucosal vaccination is thus regarded as a promising strategy to protect animals, including humans, from infections that are acquired by ingestion, inhalation or through the urogenital system. In addition, antigens delivered at the mucosal level can also elicit systemic immune responses. Therefore, mucosal vaccination is potentially effective also against systemic infections acquired through non-mucosal routes, for example, through the bite of hematophagous insects, as in the case of leishmaniasis, a widespread disease that affects humans and dogs. Here, we explored the potential of antigen rectal administration for the generation of anti-Leishmania immunity. Mice were immunized through rectal administration of whole cells of the model parasite Leishmania tarentolae (using a clone engineered to express the spike protein of the SARS-CoV-2 virus generated in a previous study). A specific anti-Leishmania IgG antibody response was detected. In addition, the recorded IgG2a/IgG1 ratio was higher than that of animals injected subcutaneously; therefore, suggesting a shift to a Th1-biased immune response. Considering the importance of a Th1 polarization as a protective response against Leishmania infections, we suggest that further investigation should be focused on the development of novel types of vaccines against these parasites based on rectal immunization.

The parasitic protozoa Leishmania (Leishmania) infantum is the etiological agent of human visceral leishmaniasis and canine leishmaniasis in South America, where Brazil is the most affected country. This zoonotic disease is transmitted by the bite of an infected phlebotomine sand fly and dogs constitute the main domestic reservoir of the parasite. In this study, we screened 2348 dogs of the municipality of Embu das Artes, Brazil, for antibodies against the parasite. Prevalence for canine leishmaniasis seropositivity was 2.81%, as assessed using a Dual-Path Platform rapid test for canine leishmaniasis. Twenty-five seropositive dogs were euthanized for parasite isolation and 14 isolates were successful obtained. Nucleotide sequencing of the internal transcribed spacer confirmed the isolates to be L. (L.) infantum, and very low sequence variability was observed among them. The in vitro susceptibility to miltefosine and paromomycin was assessed and moderate variation in paromomycin susceptibility was found among the isolates in the promastigote and intracellular amastigote stages. On the other hand, in vitro susceptibility to miltefosine of these isolates was homogenous, particularly in the amastigote stage (EC50 values from 0.69 to 2.07 μM). In addition, the miltefosine sensitivity locus was deleted in all the isolates, which does not corroborate the hypothesis that the absence of this locus is correlated with a low in vitro susceptibility. Our findings confirm that the municipality of Embu das Artes is endemic for canine leishmaniasis and that isolates from this region are susceptible to paromomycin and miltefosine, indicating the potential of these drugs to be clinically evaluated in the treatment of human visceral leishmaniasis in Brazil. [ABSTRACT FROM AUTHOR]

Simple Summary: Canine leishmaniasis (CanL) is a serious, zoonotic, protozoan disease. CanL is transmitted from infected dogs to other dogs and to humans after a blood meal taken by sandflies. Prevention mainly relies on the protection from sandfly bites. Many synthetic pyrethroids are commercially available as repellents. However, owners and practitioners are often interested in natural herbal products, which are considered safer and with low environmental impact. This case reports the lack of effectiveness of a commercial Neem-oil based product, which has long been used alone, for CanL prevention in a young French bulldog. Neem oil is a plant extract that repels insects due to one of its components, azadirachtin. Neem-based products are quite frequently used for prevention of CanL. However, data on their repellent efficacy against sandfly bites are scarce. Based on the findings of this case report, the topical application of a spray formulation of Neem oil as the sole product used for prevention of CanL is not recommended. As the spread of canine leishmaniasis (CanL) is increasing throughout the world, the need for effective agents to prevent its transmission has intensified. In this case report, an intact 1.5-year-old male French bulldog was presented for treatment of severe, sudden, and constant lameness on his right hindlimb, which had started approximately four months previously and was unresponsive to routine nonsteroidal anti-inflammatory drugs. A Neem oil-based product was sprayed three times a week on the dog's coat for about fourteen months as the only prophylactic measure against CanL. The orthopedic examination revealed grade 3–4 lameness and marked atrophy of the thigh muscles with swollen and painful right stifle joint. The radiological investigation showed polyostotic periosteal proliferation at both hindlimbs. The diagnosis of CanL was established by examination of fine-needle aspiration of lymph nodes (left prescapular, right and left popliteal) and immunofluorescence antibody testing. A leishmanicidal therapeutic protocol was prescribed. Within ten days of starting the therapy, the dog was significantly less lame, and eight months later radiographic examination revealed complete regression of the bone lesions. Some owners resort to a naturalistic approach for CanL prevention, also using products that have not been clinically evaluated. Neem oil is thought to prevent sandfly bites in dogs. Some laboratory and field studies have identified Neem oil as a possible alternative herbal drug that is repellent to sandflies. However, the clinical, laboratory, and radiographic findings clearly show that the Neem oil spray formulation used in this case report was not an effective means of CanL prevention. There is no clinical evidence in support of Neem oil-based products for the protection of dogs against CanL transmission. As Neem oil has previously been shown to be somewhat volatile, this case report suggests that even though it is a very effective repellent against sandflies, in practice, its effect on the dogs' coat was only short-lived. [ABSTRACT FROM AUTHOR]

Leishmaniasis is an important vector-borne disease that represents a serious public health problem, including in Sicily (Italy), which is considered an endemic area. We collected canine, feline and human data from 2013 to 2021 in Sicily, while entomological surveys were conducted only in 2013 and 2021. Overall, 23,794/74,349 (34.4%) of dogs and 274/4774 (11.8%) of cats were positive in one or more diagnostic tests. A total of 467 cases of human Leishmaniasis were reported, with 71% showing cutaneous and 29% visceral involvement. The provinces with the largest number of patients were Agrigento (45.4%) and Palermo (37%). In 2013, Phlebotomus perfiliewi was the dominant sandfly species in Sicily (68.7%), followed by Phlebotomus perniciosus (17.2%) and Sergentomya minuta (14%). In 2021, Phlebotomusperfiliewi was confirmed as the most common species (61.6%), followed by Phlebotomusperniciosus (33.1%) and Sergentomyaminuta (4.7%). Of particular interest was the identification of Phlebotomus papatasi (0.41%) in Agrigento. Our retrospective study can inform health authorities for the development of appropriate screening, treatment and control strategies to reduce Leishmania incidence rate. This study examined the present state of Leishmaniasis control, surveillance, and prevention in Sicily, but also highlighted deficiencies that could be addressed through the application of One-Health principles. [ABSTRACT FROM AUTHOR]

Simple Summary: Dogs are the main host of Leishmania infantum, a parasite that causes an incurable disease called canine leishmaniasis. This parasite is transmitted through the bite of a sandfly (a small insect related to mosquitoes and flies) in tropical and subtropical countries, but direct transmission between dogs, and from pregnant dogs to their puppies, exists. We reviewed the advances in tools and techniques for the surveillance of the disease, its diagnosis, treatment, and prevention. Canine leishmaniasis is expanding to the Northern Hemisphere, where it is barely known, due to climate change and the importation of dogs. Surveillance is therefore necessary in order to determine the extent of the disease in these areas and to monitor the appearance of the sandflies. Molecular techniques and rapid serological tests are now widespread for diagnosis and epidemiological studies. Several vaccines have been developed in the last decade, and even though their efficacy is limited, these advances will pave the way for the development of better vaccines against Leishmania and other parasites. Although new pharmacological tools are available, we are still waiting for the ideal drug that can eliminate the parasite from target organs and limit transmission to sandflies, without the side effects of current antileishmanials. Dog are the main reservoir of Leishmania infantum, causing canine leishmaniasis, an incurable multisystemic disease that leads to death in symptomatic dogs, when not treated. This parasite causes visceral, cutaneous, and mucosal leishmaniasis in people in the Mediterranean Basin, North Africa, South America, and West Asia. This disease is mostly unknown by veterinarians outside the endemic areas, but the disease is expanding in the Northern Hemisphere due to travel and climate change. New methodologies to study the epidemiology of the disease have found new hosts of leishmaniasis and drawn a completely new picture of the parasite biological cycle. Canine leishmaniasis diagnosis has evolved over the years through the analysis of new samples using novel molecular techniques. Given the neglected nature of leishmaniasis, progress in drug discovery is slow, and the few drugs that reach clinical stages in humans are unlikely to be commercialised for dogs, but several approaches have been developed to support chemotherapy. New-generation vaccines developed during the last decade are now widely used, along with novel prevention strategies. The implications of the epidemiology, diagnosis, treatment, and prevention of canine leishmaniasis are fundamental to public health. [ABSTRACT FROM AUTHOR]

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

Abstract Background Visceral leishmaniasis (VL) is a zoonotic disease caused by Leishmania infantum, for which dogs constitute the main urban parasite reservoir. Control measures and the treatment of canine visceral leishmaniasis (CVL) are essential to reduce VL cases. Early and accurate detection of L. infantum-infected dogs is crucial to the success of VL control. To improve the serological detection of L. infantum-exposed dogs, we evaluated the early diagnosis capacity of a recombinant protein (rLci5) in an immunosorbent assay (ELISA) to detect naturally infected dogs. Additionally, we evaluated the persistence of the positive results obtained by rLci5 ELISA in comparison to other conventional diagnostic test methods. Methods Serum samples obtained from 48 L. infantum-infected dogs involved in a cohort study were evaluated using different diagnostic methods (qPCR, EIE-LVC, DPP-LVC and splenic culture). The results were compared to rLci5 ELISA to determine its capacity to diagnose L. infantum infection at earlier infection time points. The persistence of positive diagnostic test results was also compared for each dog evaluated. Results rLci5 ELISA presented higher rates of positive results at early time points compared to the other diagnostic tests employed in the cohort study, as early as 24 months prior to detection by other tests. rLci5 ELISA positivity was 52.1% (25/48) at baseline, while qPCR was 35.4% (17/48), DPP-LVC 27.1% (13/48), EIE-LVC 22.9% (11/48) and culture only 4.2% (2/48). In at least one of the time points of the 24-month cohort study, rLci5 ELISA was positive in 100% (48/48) of the dogs, versus 83% (40/48) for qPCR, 75% (36/48) for DPP-LVC, 65% (31/48) for EIE-LVC and 31% (15/48) for culture. Investigating clinical signs in association with diagnostic test positivity, rLci5 ELISA successfully detected CVL in 62.9% (95/151) of the clinical evaluations with a score of 0–3, 64.3% (45/70) with scores between 4 and 7, and 73.7% (14/19) with scores > 7, providing higher rates of positivity than all other methods evaluated. Moreover, rLci5 ELISA presented the greatest persistence with respect to test positivity: 45.8% of the dogs evaluated. Conclusion Four diagnostic tests were compared to rLci5 ELISA, which presented earlier infection diagnosis and a greater persistence of positive test results. Accordingly, the use of the rLci5 ELISA can improve CVL diagnostic performance by detecting infected dogs sooner than other testing methods, with enhanced persistence of positive results over the course of the infection. Graphic abstract

Leishmaniasis is a life-threatening zoonosis of which dogs are the major reservoir and sandflies are the vectors. Until now, the prevalence of canine leishmaniasis (CanL) in the Slovenian dog population was unknown.

Abstract Background Leishmania parasites express various essential proteins in different growth phases (logarithmic/stationary) and forms (promastigote/amastigote). Targeting the genes encoding such proteins paves the way for controlling these parasites. Centrin is an essential gene, which its protein product seems to be vital for the proliferation of Leishmania parasites. Herein, this study was contrived to analyze the expression level of the centrin gene in different growth phases and forms of Leishmania infantum (L. infantum) parasites isolated from various endemic areas of canine leishmaniasis (CanL) in Iran. Results All three collected isolates were identified as L. infantum using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Real-time reverse transcription (RT)-PCR revealed a statistically significant up-regulation (3.13-fold) in the logarithmic phase promastigotes compared to stationary ones (p 0.05). Conclusions Centrin gene in Iranian isolates of L. infantum is more expressed in exponential than stationary phases and seems to be considered as a promising target in the development of a genetically modified live attenuated vaccine for CanL control.

• Our research investigated the genetic variations among CL, VL, and CanL strains in northern Morocco. • Molecular and genetic analyses uncovered a complex array of genetic variants in the L. infantum and L. tropica species. The number of these variants is 7 and 9 for L. tropica and L. infantum species, respectively. • The Discriminant Analysis of Principal Components (DAPC) revealed significant genetic divergences among L. infantum strains. This suggests the influence of species origin on their genetic makeup. This study investigated nine provinces in northern Morocco and collected 275 skin scraping, 22 bone marrow aspirates, and 89 fine needle aspirations from suspected cutaneous leishmaniasis (CL) and visceral leishmaniasis (VL) patients and potentially infected dogs. Molecular analysis using ITS1 RFLP PCR and RT-PCR revealed a higher prevalence of L. infantum (66.18 %; χ2 = 28.804; df = 1; P-value = 8.01e−08) than L. tropica in skin scraping, with L. infantum being the sole causative agent for both VL and canine leishmaniasis. L. infantum was predominantly found in most provinces, while L. tropica was relatively more dominant in Taza Province. Discriminant Analysis of Principal Components (DAPC) revealed distinct clustering between L. tropica and the other three species. However, no small subset of SNPs could clearly differentiate between Infantum_CL, Infantum_VL, and CanL, as they likely share a significant genetic background. The high rate of L. infantum could be attributed to the abundance of sand fly species transmitting VL. In Taza Province, Phlebotomus sergenti, responsible for anthroponotic CL, is the most abundant species. DNA sequencing demonstrated sequence heterogeneity in L. infantum (variants 1-9) and L. tropica (variants 1-7). Phylogenetic analysis showed a distinct separation between L. tropica and L. infantum strains, with an overlap among L. infantum strains isolated from cutaneous, visceral, and canine cases, and dogs serving as the central population for L. infantum. [ABSTRACT FROM AUTHOR]

Canine leishmaniasis is an important vector-borne protozoan disease in dogs that is responsible for serious deterioration in their health. In the Iberian Peninsula, as in most countries surrounding the Mediterranean Sea, canine leishmaniasis is caused by Leishmania infantum (zymodeme MON-1), a digenetic trypanosomatid that harbors in the parasitophorous vacuoles of host macrophages, causing severe lesions that can lead to death if the animals do not receive adequate treatment. Canine leishmaniasis is highly prevalent in Spain, especially in the Mediterranean coastal regions (Levante, Andalusia and the Balearic Islands), where the population of domestic dogs is very high. However, the presence of this disease has been spreading to other rural and sparsely populated latitudes, and cases of leishmaniasis have been reported for years in wildlife in northwestern Spain. This work describes for the first time the presence of wolves that tested positive for leishmaniasis in the vicinity of the Sierra de la Culebra (Zamora province, northwestern Spain), a protected sanctuary of this canid species, using PCR amplification of L. infantum DNA from different non-invasive samples such as buccal mucosa and those from both ears and hair. In addition to live animals (21), samples from carcasses of mainly roadkill animals (18) were also included and analyzed using the same technique, obtaining a positivity rate of 18 of the 39 wolves sampled (46.1%) regardless of their origin.

Background: Leishmaniases are a group of anthropo-zoonotic parasitic diseases caused by a protozoan of the Leishmania genus, affecting both humans and other vertebrates, including dogs. L. infantum is responsible for the visceral and occasionally cutaneous form of the disease in humans and canine leishmaniasis. Previously, we have shown that L. infantum induces a mild but significant increase in endoplasmic reticulum (ER) stress expression markers to promote parasites survival in human and murine infected macrophages. Moreover, we demonstrated that the miRNA hsa-miR-346, induced by the UPR-activated transcription factor sXBP1, was significantly upregulated in human macrophages infected with different L. infantum strains. However, the ER stress response in infected dogs, which represent an important reservoir for Leishmania parasite, was described once recently, whereas the miR-346 expression was not reported before. Therefore, this study aimed to investigate these pathways in the canine macrophage-like cell line DH82 infected by Leishmania spp. and to evaluate the presence of cfa-miR-346 in plasma of non-infected and infected dogs. The DH82 cells were infected with L. infantum and L. braziliensis parasites and the expression of cfa-mir-346 and several ER stress markers was evaluated by quantitative PCR (qPCR) at different time points. Furthermore, the cfa-miR-346 was monitored in plasma collected from non-infected dogs (n = 11) and dogs naturally infected by L. infantum (n = 18). Results: The results in DH82 cells showed that cfa-mir-346 was induced at both 24 h and 48 h post-infection with all Leishmania strains but not with tunicamycin, accounting for a mechanism of induction independent from sXBP1, unlike what was previously observed in human cell lines. Moreover, the cfa-miR-346 expression analysis on plasma revealed a significant increase in infected dogs compared to non-infected dogs. Conclusions: Here for the first time, we report the upregulation of cfa-miR-346 induced by Leishmania infection in canine macrophage-like cells and plasma samples of naturally infected dogs. According to our results, the cfa-miR-346 appears to be linked to infection, and understanding its role and identifying its target genes could contribute to elucidate the mechanisms underlying the host–pathogen interaction in leishmaniasis. [ABSTRACT FROM AUTHOR]

Objectives Leishmaniasis is a zoonotic disease and several drugs have been used in the treatment, including meglumine antimoniate (AME). The chemotherapy reaches clinical cure but does not eliminate parasites, contributing to drug resistance. To improve AME efficacy we incorporated it in anionic liposomes. The antiparasitic activity and intracellular localization were investigated in canine macrophages infected with Leishmania infantum. Methods Liposomes (L-AME) is composed of egg phosphatidylcholine, cholesterol, palmitoyl oleoyl phosphatidyl serine and α-tocopherol (4 : 3 : 0.4 : 0.07 mol%) plus AME. L-AME size, polydispersity, zeta potential and morphology were analysed as well as antileishmanial activity and intracellular localization in DH82 macrophages. Key findings Liposomes (360 nm) zeta potential range from −40 to −65 mV, had 23% encapsulation efficiency and were stable for 180 days at 4°C. Free AME was cytotoxic towards L. infantum infected macrophages (ID50 = 0.012 M) while L-AME did not reduce cell viability. L-AME colocalized with parasites inside macrophages in a time-dependent manner, and reduced the percentage of infected cells and the number of intracellular parasites, decreasing the infection index (75–80%) twice as compared with AME treatment. Conclusions Liposomal AME is a promising delivery system for treating visceral leishmaniasis, improving meglumine efficacy against L. infantum and minimizing its cytotoxicity towards canine macrophages [ABSTRACT FROM AUTHOR]

Abstract Background Zoonotic visceral leishmaniasis by Leishmania infantum is a first-order pathology in canine veterinary clinics 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 analyzing the impact of canine leishmaniasis (CanL) on the scientific literature, drugs or combinations used, trends in the period from 2000 to 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 = 3324) represented < 16% of all publications on leishmaniasis (n = 20,968), and of 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 (SbV), alone or in combination with allopurinol, represented > 50% of all publications on chemotherapy of CanL despite the availability of more recently marketed drugs. Conclusions Chemotherapy of CanL still relies on SbV and combinations and to a lesser extent on miltefosine (MIL). Reports on chemotherapy are scarce and 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 on the chemotherapy (etiological + symptomatic) would add value to the trials performed.

Cysteine proteinases are well-known virulence factors of Leishmania spp. with demonstrated actions in both experimental mouse infection and human infection. However, studies on these enzymes in canine leishmaniasis are scarce. Here, we show, for the first time, the reactivity of sera from dogs living in an endemic area to a recombinant protein from the COOH-terminal region of cysteine B protease. In this work, enzyme-linked immunosorbent assays were performed using a 14 kDa rcyspep protein obtained through a pET28-a expression system in Escherichia coli. First, 96-well plates were coated with rcyspep (500 ng/well) and incubated with sera from dogs (1:100). Subsequently, IgG antibody detection was performed using rabbit anti-dog IgG antibodies conjugated with peroxidase. Sera from dogs (n = 114), including suspect (n = 30) and positive (n = 50) dogs from a leishmaniasis-endemic area and dogs from a nonendemic area, (n = 34), negative for leishmaniasis, were assessed. The results showed that sera from the suspect (42%) and positive (68%) groups responded differently to the antigen titers tested above the cut-off (Optical Density = 0.166). This finding suggests that the immune response detected against cyspep may be related to clinical disorders present in these animals. Collectively, the data gathered here suggest that cyspep can sensitize the immune systems of dogs from a leishmaniasis-endemic area to elicit a humoral response, an immunological parameter indicating the contribution of this protein in host-parasite interaction.

Leishmaniasis is a heterogeneous group of neglected tropical diseases with various clinical syndromes, which is caused by obligate intracellular protozoa of the genus Leishmania and transmitted by the bite of a female phlebotomine sandfly. Humans and several animal species are considered as reservoirs of the disease. Among other animal species, dogs are the most important reservoirs in a domestic environment, maintaining the endemic focus of the parasite. The behavior of the disease progression and the clinical symptoms of the disease in the infected dog is mainly associated with depressed cellular immunity and strong humoral response. This study aimed to assess the role of Western blotting in the analysis of the idiotype expression of the two main immunoglobulins (IgG1 and IgG2) in dogs that are naturally infected with Leishmania infantum (L. infantum) and treated with N-methyl meglumine antimoniate. Interestingly, for the first time, our study identified several L. infantum antigen polypeptides (14, 31, 33, 49, 64, 66, 99, and 169 kDa) that more frequently stimulate an immune reaction in recovered dogs after treatment, whereas in the non-recovered group of dogs, four antigen polypeptides of L. infantum with molecular weights of 31, 49, 66, and 115 kDa with unfavorable prognosis were identified. Clearly, these interesting findings confirm the strong association between the detected immunodominant bands and the successful recovery in treated dogs that can be used for differentiating the treated dogs from the untreated dogs, as well as the markers of a favorable or unfavorable prognosis and, as a consequence, the prediction of the clinical outcome of the disease. Likewise, these data could be helpful in the implementation of novel vaccines from the detected antigens. [ABSTRACT FROM AUTHOR]

The detection of spatial clusters has been suggested as a potential tool for guiding the delivery of interventions against visceral leishmaniasis; however, little attention has been given to the consistency of results by using different spatial clustering methods. The present study aimed to assess the performance of three different techniques for identifying patterns in the spatial distribution of canine leishmaniasis in the city of Teresina, Brazil. This cross-sectional study was based on a serological survey for canine leishmaniasis in which each dog domicile was georeferenced to the exact location of each animal in space. The spatial analysis was performed using three methods: the Cuzick-Edwards statistic, the Hierarchical Nearest Neighbour analysis, and the Kulldorff Scan statistic. All techniques were able to identify clusters of high prevalences of canine leishmaniasis, but results were not consistent among techniques. The feasibility of the identification and location of clusters of cases in a restricted number of villages in neighborhoods might contribute to the optimization of control measures against visceral leishmaniasis. However, given the relative inconsistency of the results provided by the different methods, protocols for assessing clusters of diseases should always include more than one method of evaluation. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Purpose: Mexico is considered endemic for Leishmania; recent reports indicate autochthonous human and canine leishmaniasis caused by Leishmania mexicana in Sinaloa state. Lutzomyia sand fly are the primary vector of the parasite, although no records of phlebotomine vectors of Leishmania exist from Sinaloa. Other hematophagous dipterans, like Culicoides, could represent possible vectors of Leishmania in absence of phlebotomines. The known distribution of Culicoides includes the southern portion of Sinaloa state, in northwestern Mexico, with records of Culicoides furens. However, no studies have demonstrated the presence of Leishmania in C. furens or its possible participation in the parasite's life cycle in Mexico. This study, therefore, sought to detect DNA of Leishmania in C. furens captured in an endemic area of autochthonous canine leishmaniasis in northwestern Mexico. Methods: Culicoides were captured with CDC light traps, identified morphologically, and organized in pools. DNA was extracted, and used to amplify the ribosomal ITS1 region of Leishmania. PCR products were digested with HaeIII endonuclease; the banding patterns obtained were compared to reference strains. Results: Leishmania mexicana DNA was detected in five out of nine pools (55%) of female C. furens. Conclusion: This study offers the first evidence of L. mexicana DNA in C. furens, in an endemic area of canine leishmaniasis in northwestern Mexico, where no evidence exists of the presence of phlebotomine sand fly. [ABSTRACT FROM AUTHOR]

Background: Visceral leishmaniasis (VL) is a zoonotic disease caused by Leishmania infantum, for which dogs constitute the main urban parasite reservoir. Control measures and the treatment of canine visceral leishmaniasis (CVL) are essential to reduce VL cases. Early and accurate detection of L. infantum-infected dogs is crucial to the success of VL control. To improve the serological detection of L. infantum-exposed dogs, we evaluated the early diagnosis capacity of a recombinant protein (rLci5) in an immunosorbent assay (ELISA) to detect naturally infected dogs. Additionally, we evaluated the persistence of the positive results obtained by rLci5 ELISA in comparison to other conventional diagnostic test methods. Methods: Serum samples obtained from 48 L. infantum-infected dogs involved in a cohort study were evaluated using different diagnostic methods (qPCR, EIE-LVC, DPP-LVC and splenic culture). The results were compared to rLci5 ELISA to determine its capacity to diagnose L. infantum infection at earlier infection time points. The persistence of positive diagnostic test results was also compared for each dog evaluated. Results: rLci5 ELISA presented higher rates of positive results at early time points compared to the other diagnostic tests employed in the cohort study, as early as 24 months prior to detection by other tests. rLci5 ELISA positivity was 52.1% (25/48) at baseline, while qPCR was 35.4% (17/48), DPP-LVC 27.1% (13/48), EIE-LVC 22.9% (11/48) and culture only 4.2% (2/48). In at least one of the time points of the 24-month cohort study, rLci5 ELISA was positive in 100% (48/48) of the dogs, versus 83% (40/48) for qPCR, 75% (36/48) for DPP-LVC, 65% (31/48) for EIE-LVC and 31% (15/48) for culture. Investigating clinical signs in association with diagnostic test positivity, rLci5 ELISA successfully detected CVL in 62.9% (95/151) of the clinical evaluations with a score of 0–3, 64.3% (45/70) with scores between 4 and 7, and 73.7% (14/19) with scores > 7, providing higher rates of positivity than all other methods evaluated. Moreover, rLci5 ELISA presented the greatest persistence with respect to test positivity: 45.8% of the dogs evaluated. Conclusion: Four diagnostic tests were compared to rLci5 ELISA, which presented earlier infection diagnosis and a greater persistence of positive test results. Accordingly, the use of the rLci5 ELISA can improve CVL diagnostic performance by detecting infected dogs sooner than other testing methods, with enhanced persistence of positive results over the course of the infection. [ABSTRACT FROM AUTHOR]

Leishmaniasis is a zoonotic disease caused by Leishmania spp., impacts multiple systems and organs. While hematological and biochemical profiles aren't definitive for diagnosis, recent studies have identified the neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR) and systemic immune-inflammation index (SII) as predictors of morbidity and mortality in critically ill human and dog patients. This study examined 100 dogs diagnosed with leishmaniasis, categorized by the International Renal Interest Society (IRIS) stages 1–4. Additionally, the dogs were divided based on whether they survived less or more than one year (L1Y and G1Y). Control group consisted of 43 dogs. The NLR increased as the disease progressed (IRIS 1–4), presenting statistically significant differences (P<0.05) when compared to the control group (2,37±2,08) IRIS 3 and 4 (4,59±13,39 and 6,99±12,86, respectively), and G1Y and L1Y (3,60±4,02 and 4,87±5,82, respectively). Significant changes in SII were only evident in short-term survivors (L1Y 951,93±1402) and advanced renal disease cases (IRIS 4 stage 1073,68±1901,09). Conversely, PLR remained largely unchanged. In conclusion, these results suggest that the neutrophil-to-lymphocyte ratio (NLR) and systemic immune-inflammation index (SII) may serve as potential markers for assessing disease progression and prognosis in dogs diagnosed with leishmaniasis. • NLR values may be useful for the clinical assessment of the severity of the disease. • SII values increase with the progression of disease, being the first study that relates SII and canine leishmaniasis. • PLR as no statistically significant differences were observed against control. [ABSTRACT FROM AUTHOR]

Abstract Background Phlebotomine sand flies (Diptera: Psychodidae) are haematophagous insects that transmit the protozoan parasite Leishmania infantum (Kinetoplastida: Trypanosomatidae), the main causative agent of both zoonotic visceral leishmaniasis (VL) and canine leishmaniasis (CanL) in the Mediterranean basin. Eight species of sand flies have been previously recorded in Romania: Phlebotomus papatasi, Phlebotomus alexandri, Phlebotomus sergenti, Phlebotomus perfiliewi, Phlebotomus neglectus, Phlebotomus longiductus, Phlebotomus balcanicus and Sergentomyia minuta. Three of them (P. perfiliewi, P. neglectus and P. balcanicus) were incriminated as vectors of L. infantum. Recent reports of autochthonous CanL in Romania require updates on sand fly distribution and diversity in this country. Methods Between 2013–2014 and 2016–2018, CDC light traps and mouth aspirators were used to collect sand flies in 132 locations from Romania, indoors and around various animal species shelters. Species identification of collected specimens was done using morphological keys, genetic tools and MALDI-TOF protein profiling. Results Sand flies were present in seven localities (5.3%): Eibenthal, Baia Nouă, Gura Văii (south-western Romania, Mehedinţi County); Fundătura, Pâhneşti, Epureni (eastern Romania, Vaslui County); and Schitu (southern Romania, Giurgiu County). Of the total number of collected sand flies (n = 251), 209 (83.27%) were Phlebotomus neglectus, 39 (15.53%) P. perfiliewi, 1 (0.40%) P. papatasi, 1 (0.40%) P. balcanicus and 1 (0.40%) P. sergenti (sensu lato). Conclusions We confirmed the presence of five sand fly species previously recorded in Romania. However, their updated distribution differs from historical data. The diversity of sand fly species in Romania and their presence in areas with Mediterranean climatic influences constitutes a threat for the reemergence of vector-borne diseases. In the context of CanL and VL reemergence in Romania, but also due to imported cases of the diseases in both humans and dogs, updates on vector distribution are imperative.

Leishmaniasis is an important vector-borne disease that represents a serious public health problem, including in Sicily (Italy), which is considered an endemic area. We collected canine, feline and human data from 2013 to 2021 in Sicily, while entomological surveys were conducted only in 2013 and 2021. Overall, 23,794/74,349 (34.4%) of dogs and 274/4774 (11.8%) of cats were positive in one or more diagnostic tests. A total of 467 cases of human Leishmaniasis were reported, with 71% showing cutaneous and 29% visceral involvement. The provinces with the largest number of patients were Agrigento (45.4%) and Palermo (37%). In 2013, Phlebotomus perfiliewi was the dominant sandfly species in Sicily (68.7%), followed by Phlebotomus perniciosus (17.2%) and Sergentomya minuta (14%). In 2021, Phlebotomusperfiliewi was confirmed as the most common species (61.6%), followed by Phlebotomusperniciosus (33.1%) and Sergentomyaminuta (4.7%). Of particular interest was the identification of Phlebotomus papatasi (0.41%) in Agrigento. Our retrospective study can inform health authorities for the development of appropriate screening, treatment and control strategies to reduce Leishmania incidence rate. This study examined the present state of Leishmaniasis control, surveillance, and prevention in Sicily, but also highlighted deficiencies that could be addressed through the application of One-Health principles.