scholarly journals Leishmania infantum xenodiagnosis from vertically infected dogs reveals significant skin tropism

2021 ◽  
Vol 15 (10) ◽  
pp. e0009366
Author(s):  
Breanna M. Scorza ◽  
Kurayi G. Mahachi ◽  
Arin C. Cox ◽  
Angela J. Toepp ◽  
Adam Leal-Lima ◽  
...  
Keyword(s):  
Leishmania Infantum ◽  
Severe Disease ◽  
Clinical Status ◽  
Parasite Burden ◽  
Parasite Load ◽  
Sand Fly ◽  
Lutzomyia Longipalpis ◽  
Sand Flies ◽  
Feeding Site ◽  
Transmission Potential

Background Dogs are the primary reservoir for human visceral leishmaniasis due to Leishmania infantum. Phlebotomine sand flies maintain zoonotic transmission of parasites between dogs and humans. A subset of dogs is infected transplacentally during gestation, but at what stage of the clinical spectrum vertically infected dogs contribute to the infected sand fly pool is unknown. Methodology/Principal findings We examined infectiousness of dogs vertically infected with L. infantum from multiple clinical states to the vector Lutzomyia longipalpis using xenodiagnosis and found that vertically infected dogs were infectious to sand flies at differing rates. Dogs with mild to moderate disease showed significantly higher transmission to the vector than dogs with subclinical or severe disease. We documented a substantial parasite burden in the skin of vertically infected dogs by RT-qPCR, despite these dogs not having received intradermal parasites via sand flies. There was a highly significant correlation between skin parasite burden at the feeding site and sand fly parasite uptake. This suggests dogs with high skin parasite burden contribute the most to the infected sand fly pool. Although skin parasite load and parasitemia correlated with one another, the average parasite number detected in skin was significantly higher compared to blood in matched subjects. Thus, dermal resident parasites were infectious to sand flies from dogs without detectable parasitemia. Conclusions/Significance Together, our data implicate skin parasite burden and earlier clinical status as stronger indicators of outward transmission potential than blood parasite burden. Our studies of a population of dogs without vector transmission highlights the need to consider canine vertical transmission in surveillance and prevention strategies.

scholarly journals Leishmania infantum xenodiagnosis from vertically infected dogs reveals significant skin tropism

2021 ◽  
Author(s):  
Breanna M. Scorza ◽  
Kurayi G. Mahachi ◽  
Arin Cox ◽  
Angela J. Toepp ◽  
Adam Leal Lima ◽  
...  
Keyword(s):  
Leishmania Infantum ◽  
Severe Disease ◽  
Clinical Status ◽  
Parasite Burden ◽  
Parasite Load ◽  
Sand Fly ◽  
Lutzomyia Longipalpis ◽  
Sand Flies ◽  
Feeding Site ◽  
Transmission Potential

Background: Dogs are the primary reservoir for human visceral leishmaniasis due to Leishmania infantum. Phlebotomine sand flies maintain zoonotic transmission of parasites between dogs and humans. A subset of dogs is infected transplacentally during gestation, but at what stage of the clinical spectrum vertically infected dogs contribute to the infected sand fly pool is unknown. Methodology/Principal Findings: We examined infectiousness of dogs vertically infected with L. infantum from multiple clinical states to the vector Lutzomyia longipalpis using xenodiagnosis and found that vertically infected dogs were infectious to sand flies at differing rates. Dogs with moderate disease showed significantly higher transmission to the vector than dogs with very mild or severe disease. We documented a substantial parasite burden in the skin of vertically infected dogs by RT-qPCR, despite these dogs not having received intradermal parasites via sand flies. There was a highly significant correlation between skin parasite burden at the feeding site and sand fly parasite uptake. This suggests dogs with high skin parasite burden contribute the most to the infected sand fly pool. Although skin parasite load and parasitemia correlated with one another, the average parasite number detected in skin was significantly higher compared to blood in matched subjects. Thus, dermal resident parasites were infectious to sand flies from dogs without detectable parasitemia. Conclusions/Significance: Together, our data implicate skin parasite burden and earlier clinical status as stronger indicators of outward transmission potential than blood parasite burden. Our studies of a population of dogs without vector transmission highlights the need to consider canine vertical transmission in surveillance and prevention strategies.

scholarly journals Odour of domestic dogs infected with Leishmania infantum is attractive to female but not male sand flies: Evidence for parasite manipulation

2021 ◽  
Vol 17 (3) ◽  
pp. e1009354
Author(s):  
Monica E. Staniek ◽  
James G. C. Hamilton
Keyword(s):  
Leishmania Infantum ◽  
Etiologic Agent ◽  
Human Infection ◽  
T Test ◽  
Sand Fly ◽  
Lutzomyia Longipalpis ◽  
Sand Flies ◽  
Control Female ◽  
Behavioural Experiments ◽  
Paired T Test

Globally visceral leishmaniasis (VL) causes thousands of human deaths every year. In South America, the etiologic agent, Leishmania infantum, is transmitted from an infected canine reservoir to human hosts by the bite of the sand fly vector; predominantly Lutzomyia longipalpis. Previous evidence from model rodent systems have suggested that the odour of infected hosts is altered by the parasite making them more attractive to the vector leading to an increased biting rate and improved transmission prospects for the pathogen. However, there has been no assessment of the effect of Le infantum infection on the attractiveness of dogs, which are the natural reservoirs for human infection. Hair collected from infected and uninfected dogs residing in a VL endemic city in Brazil was entrained to collect the volatile chemical odours present in the headspace. Female and male Lu. longipalpis sand flies were offered a choice of odour entrained from infected and uninfected dogs in a series of behavioural experiments. Odour of uninfected dogs was equally attractive to male or female Lu. longipalpis when compared to a solvent control. Female Lu. longipalpis were significantly more attracted to infected dog odour than uninfected dog odour in all 15 experimental replicates (average 45.7±0.87 females attracted to infected odour; 23.9±0.82 to uninfected odour; paired T-test, P = 0.000). Male Lu. longipalpis did not significantly prefer either infected or uninfected odour (average 36.1±0.4 males to infected odour; 35.7±0.6 to uninfected odour; paired T-test, P = 0.722). A significantly greater proportion of females chose the infected dog odour compared to the males (paired T-test, P = 0.000). The results showed that the odour of dogs infected with Le. infantum was significantly more attractive to blood-seeking female sand flies than it was to male sand flies. This is strong evidence for parasite manipulation of the host odour in a natural transmission system and indicates that infected dogs may have a disproportionate significance in maintaining infection in the canine and human population.

scholarly journals Leishmania infantum Infection Modulates the Jak-STAT Pathway in Lutzomyia longipalpis LL5 Embryonic Cells and Adult Females, and Affects Parasite Growth in the Sand Fly

2021 ◽  
Vol 2 ◽  
Author(s):  
Erich Loza Telleria ◽  
Daisy Aline Azevedo-Brito ◽  
Barbora Kykalová ◽  
Bruno Tinoco-Nunes ◽  
André Nóbrega Pitaluga ◽  
...  
Keyword(s):  
Post Infection ◽  
Leishmania Infantum ◽  
Janus Kinase ◽  
Sand Fly ◽  
Lutzomyia Longipalpis ◽  
Sand Flies ◽  
Embryonic Cells ◽  
Adult Females ◽  
Parasite Survival ◽  
Stat Pathway

Phlebotomine sand flies (Diptera, Psychodidae) belonging to the Lutzomyia genus transmit zoonoses in the New World. Lutzomyia longipalpis is the main vector of Leishmania infantum, which is the causative agent of visceral leishmaniasis in Brazil. To identify key molecular aspects involved in the interaction between vector and pathogens and contribute to developing disease transmission controls, we investigated the sand fly innate immunity mediated by the Janus kinase/signal transducer and activator of transcription (Jak-STAT) pathway in response to L. infantum infection. We used two study models: L. longipalpis LL5 embryonic cells co-cultured with L. infantum and sand fly females artificially infected with the parasite. We used qPCR to follow the L. longipalpis gene expression of molecules involved in the Jak-STAT pathway. Also, we modulated the Jak-STAT mediated immune response to understand its role in Leishmania parasite infection. For that, we used RNAi to silence the pathway regulators, protein inhibitor of activated STATs (PIAS) in LL5 cells, and STAT in adult females. In addition, the pathway suppression effect on parasite development within the vector was assessed by light microscopy in late-phase infection. The silencing of the repressor PIAS in LL5 cells led to a moderate increase in a protein tyrosine phosphatase 61F (PTP61F) expression. It suggests a compensatory regulation between these two repressors. L. infantum co-culture with LL5 cells upregulated repressors PIAS, suppressor of cytokine signaling (SOCS), and PTP61F. It also downmodulated virus-induced RNA-1 (VIR-1), a pathway effector, indicating that the parasite could repress the Jak-STAT pathway in LL5 cells. In Leishmania-infected L. longipalpis females, STAT and the antimicrobial peptide attacin were downregulated on the third day post-infection, suggesting a correlation that favors the parasite survival at the end of blood digestion in the sand fly. The antibiotic treatment of infected females showed that the reduction of gut bacteria had little effect on the Jak-STAT pathway regulation. STAT gene silencing mediated by RNAi reduced the expression of inducible nitric oxide synthase (iNOS) and favored Leishmania growth in sand flies on the first day post-infection. These results indicate that STAT participated in the iNOS regulation with subsequent effect on parasite survival.

scholarly journals Transmission Potential of Antimony-Resistant Leishmania Field Isolates

2014 ◽  
Vol 58 (10) ◽  
pp. 6273-6276 ◽  
Author(s):  
Veronika Seblova ◽  
Bruno Oury ◽  
Naouel Eddaikra ◽  
Khatima Aït-Oudhia ◽  
Francine Pratlong ◽  
...  
Keyword(s):  
Late Stage ◽  
Leishmania Infantum ◽  
Lutzomyia Longipalpis ◽  
Parasite Transmission ◽  
Sand Flies ◽  
Content Type ◽  
Phlebotomus Perniciosus ◽  
Transmission Potential ◽  
Resistant Phenotype ◽  
Resistant Strains

ABSTRACTWe studied the development of antimony-resistantLeishmania infantumin natural vectorsLutzomyia longipalpisandPhlebotomus perniciosusto ascertain the risk of parasite transmission by sand flies. All three resistant strains produced fully mature late-stage infections in sand flies; moreover, the resistant phenotype was maintained after the passage through the vector. These results highlight the risk of circulation of resistantLeishmaniastrains and question the use of human drugs for treatment of dogs asLeishmaniareservoirs.

scholarly journals Insecticide-impregnated netting as a potential surface treatment: an alternative to insecticide spraying for control of the leishmaniasis vector Lutzomyia longipalpis (Diptera: Psychodidae)?

2020 ◽  
Author(s):  
Vanessa Barbosa ◽  
Cristian F Souza ◽  
Derek Gatherer ◽  
Reginaldo P Brazil ◽  
James Gordon Campbell Hamilton
Keyword(s):  
Aggregation Pheromone ◽  
Leishmania Infantum ◽  
Intervention Studies ◽  
Potential Surface ◽  
Lower Proportion ◽  
Sand Fly ◽  
Lutzomyia Longipalpis ◽  
Sand Flies ◽  
Synthetic Pheromone ◽  
Canine Infection

Abstract Background: The sand fly, Lutzomyia longipalpis, is the main vector of Leishmania infantum in Brazil. A previous laboratory study showed that covering surfaces with insecticide-impregnated netting may provide an alternative method for killing sand flies. Synthetic male Lu. longipalpis sex/aggregation pheromone co-located with micro-encapsulated l-cyhalothrin demonstrated the potential of “lure-and-kill” to significantly reduce canine infection and sand fly densities. In this study we were interested to determine if insecticide impregnated netting could replace sprayed insecticide for Lu. longipalpis control.Methods: We placed synthetic pheromone in experimental and real chicken sheds treated with a 1m2 surface of either sprayed insecticide or insecticide-impregnated netting. Two experiments in experimental chicken sheds were carried out to determine the effect of the insecticide treatments on Lu. longipalpis over 1-week and 16-week periods. We counted the number of Lu. longipalpis collected overnight and dead at 24 hours. Two longitudinal intervention studies were carried in real chicken sheds and compared the numbers of Lu. longipalpis (collected and dead at 24h) before adding the intervention (either the netting or sprayed insecticide treatments) with the numbers collected 24h after the intervention. Results: In the first experiment all flies caught in the spray treated experimental chicken sheds were dead at 24 hours and in netting treated sheds 97% of females and 88% of males were dead at 24 hours (257 vs 225, Wilcoxon Signed Ranks Test P=0.043). The netting and spray treated traps were equally effective at killing both female and male Lu. longipalpis over the first 8-weeks however after 16-weeks both treatments killed a significantly lower proportion of females (64%vs 96%; P=0.000) and males 89%vs 100%; P=0.000) compared to the beginning. In the first of the longitudinal studies in real chicken sheds only the netting intervention significantly increased the proportion of females dead after 24h (60%vs81%; P=0.042). The subsequent study showed that both netting and spraying treatments had similarly significant impacts on the proportion of females dead after 24h (netting: 60%vs80%: P=0.0194 and spraying: 43%vs72%: P=0.0004).Conclusions: The netting and spray insecticide interventions (with synthetic sex/aggregation pheromone) have similar impacts on the Lu. longipalpis population.

Transmission potential of paromomycin-resistant Leishmania infantum and Leishmania donovani

2019 ◽  
Vol 75 (4) ◽  
pp. 951-957 ◽  
Author(s):  
S Hendrickx ◽  
L Van Bockstal ◽  
H Aslan ◽  
J Sadlova ◽  
L Maes ◽  
...  
Keyword(s):  
Leishmania Infantum ◽  
Leishmania Donovani ◽  
Leishmania Species ◽  
Blood Feeding ◽  
Sand Fly ◽  
Lutzomyia Longipalpis ◽  
Transmission Potential ◽  
Laboratory Rodents ◽  
The Impact

Abstract Objectives Former studies demonstrated quick selection of paromomycin resistance for Leishmania infantum and Leishmania donovani accompanied by increased fitness. The present study aimed to interpret these findings in an epidemiological context by comparing infection of WT and experimentally derived paromomycin-resistant strains in the sand fly vector. Methods Depending on the Leishmania species, Lutzomyia longipalpis and Phlebotomus perniciosus or Phlebotomus argentipes sand flies were artificially infected with procyclic promastigotes of WT and paromomycin-resistant L. infantum (MHOM/FR/96/LEM3323-cl4) or L. donovani (MHOM/NP/03/BPK275/0-cl18). The infection rate and gut/stomodeal valve colonization were determined to monitor parasite phenotypic behaviour within the vector. The impact of the previously described gain of fitness in the vertebrate host on infectivity for the vector was assessed by feeding L. longipalpis on Syrian golden hamsters heavily infected with either WT or paromomycin-resistant parasites. Results WT and paromomycin-resistant Leishmania of both species behaved similarly in terms of infection and parasite location within the studied sand fly species. Blood feeding on infected hamsters did not reveal differences in acquisition of WT and paromomycin-resistant parasites, despite the higher organ burdens observed for the paromomycin-resistant strain. Strains remained resistant after passage in the vector. Conclusions Although paromomycin-resistant parasites show an increased parasite fitness in vitro and in laboratory rodents, the intrinsic infection potential of paromomycin-resistant parasites remains unaltered in the sand fly. Of importance is the fact that paromomycin-resistant Leishmania are able to complete development in the natural vectors and produce stomodeal infection with metacyclic forms, which clearly suggests their potential to spread and circulate in nature.

scholarly journals Binding of Leishmania infantum Lipophosphoglycan to the Midgut Is Not Sufficient To Define Vector Competence in Lutzomyia longipalpis Sand Flies

mSphere ◽  
2020 ◽  
Vol 5 (5) ◽  
Author(s):  
Iliano V. Coutinho-Abreu ◽  
James Oristian ◽  
Waldionê de Castro ◽  
Timothy R. Wilson ◽  
Claudio Meneses ◽  
...  
Keyword(s):  
Leishmania Infantum ◽  
Vector Competence ◽  
Sand Fly ◽  
Parasite Development ◽  
Lutzomyia Longipalpis ◽  
Sand Flies ◽  
Content Type ◽  
Survival And Development ◽  
Blood Meal Digestion ◽  
Insect Gut

It is well established that the presence of LPG is sufficient to define the vector competence of restrictive sand fly vectors with respect to Leishmania parasites. However, the permissiveness of other sand flies with respect to multiple Leishmania species suggests that other factors might define vector competence for these vectors. In this study, we investigated the underpinnings of Leishmania infantum survival and development in its natural vector, Lutzomyia longipalpis. We found that LPG-mediated midgut binding persists in late-stage parasites. This observation is of relevance for the understanding of vector-parasite molecular interactions and suggests that only a subset of infective metacyclic-stage parasites (metacyclics) lose their ability to attach to the midgut, with implications for parasite transmission dynamics. However, our data also demonstrate that LPG is not a determining factor in Leishmania infantum retention in the midgut of Lutzomyia longipalpis, a permissive vector. Rather, LPG appears to be more important in protecting some parasite strains from the toxic environment generated during blood meal digestion in the insect gut. Thus, the relevance of LPG in parasite development in permissive vectors appears to be a complex issue and should be investigated on a strain-specific basis.

scholarly journals The sand fly (Diptera: Psychodidae) fauna of the urban area of Lassance, Northeast Minas Gerais, Brazil

PLoS ONE ◽  
2021 ◽  
Vol 16 (10) ◽  
pp. e0257043
Author(s):  
Gabriel Barbosa Tonelli ◽  
Camila Binder ◽  
Victoria Laporte Carneiro Nogueira ◽  
Marina Henriques Prado ◽  
Gabriela Gonçalves Theobaldo ◽  
...  
Keyword(s):  
Leishmania Infantum ◽  
Abundant Species ◽  
Minas Gerais ◽  
Sand Fly ◽  
Canine Leishmaniasis ◽  
Lutzomyia Longipalpis ◽  
Scaling Analysis ◽  
Sand Flies ◽  
Risk Area ◽  
Risk Areas

The present study aimed to check the sand flies’ fauna on the municipality of Lassance, Minas Gerais, Brazil and detect the presence of Leishmania DNA on the female captured and determine the risk areas of the municipality. Sand flies were collected monthly from May 2018 to April 2019 using automatic light traps for 3 consecutive nights. Eight houses were selected as sample points due its previous reports of tegumentary leishmaniasis and/or canine leishmaniasis. The sand fly’s fauna found on the present study it’s represented by several medical importance species and the most abundant species found were Lutzomyia longipalpis (77.09%) and Nyssomyia intermedia (10.06%). Leishmania infantum DNA was detected in a pool of Lu. longipalpis resulting on a 2.81% of infection rate. By the frequency of the two most abundant species on this study, we developed a risk area map and it draws attention to sample point 6 due to disparate abundance of sand flies at this site (81.81%). Statistical overview shows Lu. longipalpis as dominant species and, still, Non-Metric Multidimensional Scaling analysis reveal high similarity on fauna’s diversity on the study area. Our findings suggest that the diversity of sand flies from the municipality of Lassance may promote the circulation of Leishmania infantum parasites putting in risk the habitants and other mammal’s species. Still, our study reinforces the necessity of specific studies focused on breed sites of phlebotomine and its’ ecology to expand the knowledge about the behaviour of this group of insects applying directly to leishmaniases’ epidemiology.

scholarly journals Binding of Leishmania infantum LPG to the midgut is not sufficient to define vector competence in Lutzomyia longipalpis sand flies

2020 ◽  
Author(s):  
Iliano V. Coutinho-Abreu ◽  
James Oristian ◽  
Waldionê de Castro ◽  
Timothy R. Wilson ◽  
Claudio Meneses ◽  
...  
Keyword(s):  
Late Stage ◽  
Leishmania Infantum ◽  
Vector Competence ◽  
Sand Fly ◽  
Parasite Development ◽  
Lutzomyia Longipalpis ◽  
Sand Flies ◽  
Survival And Development ◽  
Sand Fly Vectors

AbstractThe major surface lipophosphoglycan (LPG) of Leishmania parasites is critical to vector competence in restrictive sand fly vectors by mediating Leishmania attachment to the midgut epithelium, considered essential to parasite survival and development. However, the relevance of LPG for sand flies that harbor multiple species of Leishmania remains elusive. We tested binding of Leishmania infantum wild type (WT), LPG-defective (Δlpg1 mutants) and add-back lines (Δlpg1 + LPG1) to sand fly midguts in vitro and their survival in Lutzomyia longipalpis sand flies in vivo. Le. infantum WT parasites attached to the Lu. longipalpis midgut in vitro with late-stage parasites binding to midguts in significantly higher numbers compared to early-stage stage promastigotes. Δlpg1 mutants did not bind to Lu. longipalpis midguts, and this was rescued in the Δlpg1 + LPG1 lines, indicating that midgut binding is mediated by LPG. When Lu. longipalpis sand flies were infected with either Le. infantum WT, Δlpg1, or Δlpg1 + LPG1 of the BH46 or BA262 strains, the BH46 Δlpg1 mutant, but not the BA262 Δlpg1 mutant, survived and grew to similar numbers as the WT and Δlpg1 + LPG1 lines. Exposure of BH46 and BA262 Δlpg1 mutants to blood engorged midgut extracts led to the mortality of the BA262 Δlpg1 but not the BH46 Δlpg1 parasites. These findings suggest that Le. infantum LPG protects parasites on a strain-specific basis early in infection, likely against toxic components of blood digestion, however, it is not necessary to prevent Le. infantum evacuation along with the feces in the permissive vector Lu. longipalpis.IMPORTANCEIt is well established that LPG is sufficient to define the vector competence of restrictive sand fly vectors to Leishmania parasites. However, the permissiveness of other sand flies to multiple Leishmania species suggests that other factors might define vector competence for these vectors. In this study, we investigated the underpinnings of Leishmania infantum survival and development in its natural vector Lutzomyia longipalpis. We found out that LPG-mediated midgut binding persists in late-stage parasites. This observation is paradigm-changing and suggests that only a subset of infective metacyclics lose their ability to attach to the midgut with implications for parasite transmission dynamics. However, our data also demonstrate that LPG is not a determining factor in Leishmania infantum retention in the midgut of Lutzomyia longipalpis, a permissive vector. Rather, LPG appears to be more important in protecting some parasite strains from the toxic environment generated during blood meal digestion in the insect gut. Thus, the relevance of LPG in parasite development in permissive vectors appears to be complex and should be investigated on a strain-specific basis.

Detection of Pintomyia fischeri (Diptera: Psychodidae) With Leishmania infantum (Trypanosomatida: Trypanosomatidae) Promastigotes in a Focus of Visceral Leishmaniasis in Brazil

2020 ◽  
Author(s):  
Fredy Galvis-Ovallos ◽  
Adriele Eiko Ueta ◽  
Gabriella de Oliveira Marques ◽  
Ana Maria Casagrande Sarmento ◽  
Gabriela Araujo ◽  
...  
Keyword(s):  
Visceral Leishmaniasis ◽  
Leishmania Infantum ◽  
Species Complex ◽  
Natural Infection ◽  
Metropolitan Region ◽  
Sand Fly ◽  
Surveillance Program ◽  
Lutzomyia Longipalpis ◽  
Sand Flies ◽  
Vector Surveillance

Abstract Visceral leishmaniasis is spreading in Brazil where the main vector of its agent, Leishmania infantum Nicolle, 1908, is the Lutzomyia longipalpis (Lutz & Neiva, 1912) species complex (Diptera: Psychodidae: Phlebotominae), on which many of the activities of the visceral leishmaniasis surveillance program are based. However, there are areas where canine, and/or human cases have been occurring without the presence of this species complex as in the western part of the Greater São Paulo Metropolitan region, where Embu das Artes municipality is situated. In this area, Pintomyia fischeri (Pinto, 1926) has been implicated as potential vector of Le. infantum but so far its natural infection with this parasite has not yet been ascertained. Therefore, the present study sought to investigate the natural infection in sand flies of a CVL focus in Embu das Artes. The sand fly collections were undertaken with Shannon and CDC traps, monthly, between 1800 and 2100 hours from November 2018 to October 2019, inclusive. A total of 951 sand flies (457 males and 494 females), belonging to 10 species, were captured. Pintomyia fischeri was the predominant species (89.5%); of which 426 females were dissected and one of them (0.23%) was found to be harboring flagellates in its midgut. A sample of these flagellates was isolated in culture and characterized by a 234 base pair fragment of Leishmania heat-shock protein 70 gene (hsp70) and restriction fragment length polymorphism with Hae III restriction enzyme as Le. infantum. This finding reinforces previous evidence of Pi. fischeri as a vector of Le. infantum in foci of visceral leishmaniasis and highlights the importance of vector surveillance in areas where this species occurs.

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