Detection of <i>Leishmania (L.) infantum</i> in stray dogs by molecular techniques with sensitive species-specific primers

Silva, Rodrigo da; Richini-Pereira, Virgínia Bodelão; Kikuti, Mariana; Marson, Pâmela Merlo; Langoni, Hélio

doi:10.1080/01652176.2016.1252073

Cited by 20 publications

(16 citation statements)

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“…Among them, the mini exon/spliced leader gene (Azmi, Nasereddin, Ereqat, Schönian, & Abdeen, 2010;Marfurt, Niederwieser, Makia, Beck, & Felger, 2003), the hsp70 gene (Montalvo, Fraga, Maes, Dujardin, & Van Der Auwera, 2012), the ribosomal internal transcribed spacer (ITS1-2) regions (de Almeida et al, 2011;Del Río et al, 2014;Schönian et al, 2003) and the kinetoplast, mitochondrial DNA (kDNA) (Millán et al, 2011;Souza Castro et al, 2018) have enough sequence polymorphisms to allow sensitive detection of genetic variability within Leishmania species (Akhoundi et al, 2017). Their high copy number and the presence of variable and conserved DNA regions make them ideal for pathogen detection (high sensitivity) and typing (variability studies) (de Almeida et al, 2011;Schönian et al, 2003;Silva, Richini-Pereira, Kikuti, Marson, & Langoni, 2017). Intraspecific variability was recorded in the internal transcribed spacer 2 (ITS2) sequence of L. infantum from humans, dogs and wildlife from Spain (Del Río et al, 2014).…”

Section: Introductionmentioning

confidence: 99%

Genetic diversity and phylogenetic relationships between Leishmania infantum from dogs, humans and wildlife in south‐east Spain

Ortuño

Latrofa

Iborra

et al. 2019

Zoonoses and Public Health

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Leishmania infantum causes human and canine leishmaniosis. The parasite, transmitted by phlebotomine sand flies, infects species other than dogs and people, including wildlife, although their role as reservoirs of infection remains unknown for most species. Molecular typing of parasites to investigate genetic variability and evolutionary proximity can help understand transmission cycles and designing control strategies. We investigated Leishmania DNA variability in kinetoplast (kDNA) and internal transcribed spacer 2 (ITS2) sequences in asymptomatically infected wildlife (n = 58) and symptomatically and asymptomatically infected humans (n = 38) and dogs (n = 15) from south‐east Spain, using single nucleotide polymorphisms (SNPs) and in silico restriction fragment length polymorphism (RFLP) analyses. All ITS2 sequences (n = 76) displayed a 99%–100% nucleotide identity with a L. infantum reference sequence, except one with a 98% identity to a reference Leishmania panamensis sequence, from an Ecuadorian patient. No heterogeneity was recorded in the 73 L. infantum ITS2 sequences except for one SNP in a human parasite sequence. In contrast, kDNA analysis of 44 L. infantum sequences revealed 11 SNP genotypes (nucleotide variability up to 4.3%) and four RFLP genotypes including B, F and newly described S and T genotypes. Genotype frequency was significantly greater in symptomatic compared to asymptomatic individuals. Both methods similarly grouped parasites as predominantly or exclusively found in humans, in dogs, in wildlife or in all three of them. Accordingly, the phylogenetic analysis of kDNA sequences revealed three main clusters, two as a paraphyletic human parasites clade and a third including dogs, people and wildlife parasites. Results suggest that Leishmania infantum genetics is complex even in small geographical areas and that, probably, several independent transmission cycles take place simultaneously including some connecting animals and humans. Investigating these transmission networks may be useful in understanding the transmission dynamics, infection risk and therefore in planning L. infantum control strategies.

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Section: Introductionmentioning

confidence: 99%

Genetic diversity and phylogenetic relationships between Leishmania infantum from dogs, humans and wildlife in south‐east Spain

Ortuño

Latrofa

Iborra

et al. 2019

Zoonoses and Public Health

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“…In this way, we concluded that it was not feasible to perform screening with nonspecific primers for the diagnosis of Leishmania infection in equines, since not all positive animals were infected by the genus Leishmania. The specificity of these PCR primers was tested by their inability to amplify DNA from pathogens such as T. cruzi, E. canis, B. canis, and T. gondii (Almeida et al, 2013;Le Fichoux et al, 1999;R. C. Silva et al, 2017;Solcà et al, 2012).…”

Section: Discussionmentioning

confidence: 99%

Applications of polymerase chain reaction for the detection of equine Leishmania sp. infection

Escobar

Döwich

Cantele

et al. 2020

SCA

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Leishmaniasis is a neglected zoonotic disease caused by a variety of pathogenic Leishmania species. In the New World, especially in Brazil, visceral leishmaniasis (VL) is caused by Leishmania infantum. The pathogen can infect several animal species including dogs, foxes, rodents, primates, felines, equines and humans. Dogs act as the primary domestic reservoirs. This study aimed to use polymerase chain reaction (PCR) for detecting Leishmania infection in horses living in a canine visceral leishmaniasis (CVL) endemic region. DNA samples from horse peripheral blood were used to perform PCR. Templates were amplified using primers for the kinetoplast DNA (kDNA) minicircles, which were able to detect different species of Leishmania. In addition, primers for internal transcribed spacer (ITS) of ribosomal DNA were used for detection of Trypanosomatidae sp. Amongst the 75 (39%) positive PCR samples from total 192 samples, 21 samples were positive for kDNA and 63 samples were positive for either ITS, ITS1, or ITS2 gene markers. The kDNA PCR and sequencing allowed the detection of L. infantum in horse blood samples. To our knowledge, this is the first report of equine infection with L. infantum in Southern Brazil. These results proved that L. infantum could also infect horses in addition to humans and dogs, as well as in European countries. This conclusion emphasizes the urgent need to follow up investigation of the infection in these animals.

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“…Thermal cycling conditions followed those of El Tai et al (2000) [28] and Silva et al [29]. The amplified products were identified by 1.5% agarose gel electrophoresis containing 1.0 μL/10 mL of SYBR safe DNA gel stain (Invitrogen, Life Technologies, USA).…”

Section: Methodsmentioning

confidence: 99%

Visceral leishmaniasis in an environmentally protected area in southeastern Brazil: Epidemiological and laboratory cross-sectional investigation of phlebotomine fauna, wild hosts and canine cases

et al. 2017

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BackgroundLeishmaniasis is a rapidly expanding zoonosis that shows increasing urbanization. Concern exists regarding the role of wildlife in visceral leishmaniasis (VL) transmission, due to frequent natural or anthropogenic environmental changes that facilitate contact between wildlife, humans and their pets. The municipality of Campinas, in southeastern Brazil, initially recorded VL in 2009, when the first autochthonous case was confirmed in a dog living in an upscale residential condominium, located inside an environmentally protected area (EPA). Since then, disease transmission remains restricted to dogs inhabiting two geographically contiguous condominiums within the EPA.Methodology/Principal findingsWe conducted a cross-sectional study of the VL focus to investigate Leishmania spp. infection in domestic dogs, wild mammals and sand flies using molecular tools and recommended serological techniques. Canine seroprevalences of 1.5% and 1.2% were observed in 2013 and 2015, respectively. Six insect species, confirmed or suspected vectors or potential transmitters of Leishmania, were identified. Two specimens of the main L. (L.) infantum vector in Brazil, Lutzomyia longipalpis, were captured in the EPA. Natural infection by L. (L.) infantum was recorded in one Expapillata firmatoi specimen and two Pintomyia monticola. Natural infection by L. (L.) infantum and Leishmania subgenus Viannia was also detected in two white-eared opossums (Didelphis albiventris), a known reservoir of VL. Geographical coordinates of each sampling of infected animals were plotted on a map of the EPA, demonstrating proximity between these animals, human residences, including the dogs positive for VL, and forest areas.Conclusions/SignificanceThe EPA, which is inhabited by humans, has an active VL focus. The risk of establishing and maintaining disease transmission foci in similar scenarios, i.e. wild areas that undergo environmental modifications, is evident. Moreover, different epidemiological profiles of VL must be included to elaborate prevention and control measures that consider the particularities of each transmission area.

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Detection of Leishmania (L.) infantum in stray dogs by molecular techniques with sensitive species-specific primers

Cited by 20 publications

References 35 publications

Genetic diversity and phylogenetic relationships between Leishmania infantum from dogs, humans and wildlife in south‐east Spain

Genetic diversity and phylogenetic relationships between Leishmania infantum from dogs, humans and wildlife in south‐east Spain

Applications of polymerase chain reaction for the detection of equine Leishmania sp. infection

Visceral leishmaniasis in an environmentally protected area in southeastern Brazil: Epidemiological and laboratory cross-sectional investigation of phlebotomine fauna, wild hosts and canine cases

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