Human Endemic Areas Research Articles

Spatial-temporal distribution of visceral leishmaniasis in Rio de Janeiro, Brazil, 2001-2020: expansion and challenges.

Visceral leishmaniasis results from complex interactions among humans, dogs and environment. Brazil accounts for 97% of cases in the Americas. Twenty years (2001-2020) of the endemic disease in the state of Rio de Janeiro were studied. Incidence, lethality, sociodemographic and clinical characteristics were investigated, complemented with spatial methodologies (kernel and clusters). Ninety-seven human cases and 625 dogs were reported. Of the 92 cities, 22 were human endemic areas. The state had a low incidence level (0.6 per 100 000). Lethality was higher compared with the Brazilian average. More than 90% of infections occurred in urban areas. Most cases (66%) occurred in men. The predominant age groups were 0-4 y (28.7%) and 20-39 y (32.9%). Fever (89.5%), splenomegaly (83.2%) and hepatomegaly (76.8%) were the main clinical manifestations. Spatial analysis showed a displacement of the human endemic: in the first decade (2001-2010), cases were concentrated in the Metropolitan region, and in the second decade (2011-2020) in the Médio Paraíba region of the state. Most of the endemic area (56.4%) had canine infections without reported human cases. Disorderly urbanisation and precarious living conditions favour the transmission of the disease. Changes in the environment and migratory processes contribute to its expansion.

High prevalence of Schistosoma japonicum by perfusion in naturally exposed water buffalo in a region of the Philippines endemic for human schistosomiasis.

In the past decade, ecological surveys emphasized rats and dogs as the most significant animal reservoirs for Schistosoma japonicum (S.j) in the Philippines. However, recent studies demonstrated 51–91% prevalence of schistosomiasis among water buffalo using qPCR in the Sj endemic regions in the Philippines. In order to resolve the inconsistency of reported surveys regarding Sj endemicity among carabao, a domestic water buffalo that is the most important draught animal, we introduced 42 schistosome negative water buffalo to Macanip, Jaro municipality, Leyte, the Philippines, a subsistence rice-farming village that has been the focus of schistosomiasis japonica studies of our group for the past 20 years. We conducted perfusion to the remaining 34 buffalo that survived 10 months of nature exposure and Typhoon Haiyan. Thirty-three water buffalo were found to be positive with at least 1 pair of worms from the mesenteric vein. The infection rate is 97%, with the worm burden of 94 (95% confidence interval, 49–138 worms) worms. To our knowledge, this is the first report about S. japonicum worm burden in naturally infected water buffalo in the Philippines. The fact that with less than one-year of exposure, in this human schistosomiasis endemic area, only 1 out of 34 water buffalo was uninfected is striking. Urgent attention is needed for a cost-effective technique for monitoring Sj infection in animals and humans. Meanwhile, intervention implementation, including water buffalo treatment and vaccination, should be taken into consideration.

Spread of the fascioliasis endemic area assessed by seasonal follow-up of rDNA ITS-2 sequenced lymnaeid populations in Cajamarca, Peru

Fascioliasis is a worldwide emerging snail-borne zoonotic trematodiasis with a great spreading capacity linked to animal and human movements, climate change, and anthropogenic modifications of freshwater environments. South America is the continent with more human endemic areas caused by Fasciola hepatica, mainly in high altitude areas of Andean regions. The Peruvian Cajamarca area presents the highest human prevalences reported, only lower than those in the Bolivian Altiplano. Sequencing of the complete rDNA ITS-2 allowed for the specific and haplotype classification of lymnaeid snails collected in seasonal field surveys along a transect including 2007–3473 m altitudes. The species Galba truncatula (one haplotype preferentially in higher altitudes) and Pseudosuccinea columella (one haplotype in an isolated population), and the non-transmitting species Lymnaea schirazensis (two haplotypes mainly in lower altitudes) were found. Climatic seasonality proved to influence G. truncatula populations in temporarily dried habitats, whereas L. schirazensis appeared to be more climatologically independent due to its extreme amphibious ecology. Along the southeastern transect from Cajamarca city, G. truncatula and L. schirazensis shared the same site in 7 localities (46.7% of the water collections studied). The detection of G. truncatula in 11 new foci (73.3%), predominantly in northern localities closer to the city, demonstrate that the Cajamarca transmission risk area is markedly wider than previously considered. Lymnaea schirazensis progressively increases its presence when moving away from the city. Results highlight the usefulness of lymnaeid surveys to assess borders of the endemic area and inner distribution of transmission foci. Similar lymnaeid surveys are still in need to be performed in the wide northern and western zones of the Cajamarca city. The coexistence of more than one lymnaeid transmitting species, together with a morphologically indistinguishable non-transmitting species and livestock movements inside the area, conform a complex scenario which poses difficulties for the needed One Health control intervention.

Domestic pig prioritized in one health action against fascioliasis in human endemic areas: Experimental assessment of transmission capacity and epidemiological evaluation of reservoir role

The Northern Bolivian Altiplano is the human fascioliasis hyperendemic area where the highest prevalences and intensities in humans have been reported. Preventive chemotherapy was implemented in the last ten years. Surveillance showed high human infection and re-infection rates in between the annual triclabendazole monodose treatments. A complementary One Health control action was launched to decrease the infection risk. Among the multidisciplinary axes, there is the need to establish animal reservoir species priorities for a more efficient control. Laboratory and field studies were performed for the first time to assess the Fasciola hepatica transmission capacity of the pig and its potential reservoir role. The experimental follow-up of altiplanic pig isolates through altiplanic Galba truncatula snail vector isolates were performed at different miracidial doses and different day/night temperatures. Experiments included egg embryonation, miracidial infectivity, lymnaeid snail infection, intramolluscan larval development, cercarial production, chronobiology of the cercarial shedding, vector survival to infection, metacercarial infectivity of mammal host, and adult stage development. Surveys included the assessment of prevalence, intensity, egg measurements and egg shedding rates in nature. Pig contribution was evaluated by comparing with the main altiplanic reservoirs sheep and cattle. Results demonstrated that the pig assures the whole F. hepatica life cycle and participates in its transmission in this area. The fast egg embryonation, high cercarial production, long multi-wave shedding chronobiological pattern in monomiracidial infections at permanent 20 °C temperature, and the high daily egg outputs per pig are worth mentioning. The high infection risk suggests early infection of freely running piglets and evolutionary long-term adaptation of the liver fluke to this omnivorous mammal, despite its previously evoked resistance or non-suitability. Genetic, physiological and immune similarities with humans may also underlie the parasite adaptation to humans in this area. The pig should be accordingly included for appropriate control measures within a One Health action against human fascioliasis. The pig should henceforth be considered in epidemiological studies and control initiatives not only in fascioliasis endemic areas with human infection risk on other Andean countries, but also in rural areas of Latin America, Africa and Asia where domestic pigs are allowed to run freely.

Sheep and Cattle Reservoirs in the Highest Human Fascioliasis Hyperendemic Area: Experimental Transmission Capacity, Field Epidemiology, and Control Within a One Health Initiative in Bolivia.

The Northern Bolivian Altiplano is the human fascioliasis hyperendemic area where the highest prevalences and intensities of infection by Fasciola hepatica in humans have been reported. Four animal species are the reservoir species for F. hepatica in this area, namely, sheep, cattle, pigs, and donkeys. Livestock for the Aymara inhabitants is crucial because vegetable cultures are not viable due to the inhospitality of the very high altitude of 3,820–4,100 m. A One Health initiative has been implemented in this area in recent years, as the first such control action in a human endemic area ever. Among the different control axes included, special focus is devoted to the two main reservoirs sheep and cattle. Egg embryonation, miracidial infectivity, intramolluscan development, cercarial production, infected snail survival, and metacercarial infectivity were experimentally studied in altiplanic sheep and cattle isolates. These laboratory studies were performed using altiplanic isolates of the lymnaeid species Galba truncatula, the only vector present in the hyperendemic area. Experiments were made at constant 12 h day/12 h night and varying 20/20°C and 22/5°C photoperiods. Infections were implemented using mono-, bi-, and trimiracidial doses. Results demonstrate that sheep and cattle have the capacity to assure F. hepatica transmission in this very high-altitude area. Field surveys included prevalence studies by coprology on fecal samples from 1,202 sheep and 2,690 cattle collected from different zones of the Northern Bolivian Altiplano. Prevalences were pronouncedly higher and more homogeneous in sheep (63.1%; range: 38.9–68.5%) than in cattle (20.6%; range: 8.2–43.3%) in each one of the different zones. Although similarities between the prevalences in sheep and cattle appeared in the zones of the highest and lowest infection rates, this disappeared in the other zones due to cattle treatments. Comparison with past surveys demonstrates that this hyperendemic area is stable from the disease transmission point of view. Therefore, the control design should prioritize sheep and cattle within the One Health action. Studies performed in the Bolivian Altiplano furnish a baseline for future initiatives to assess the transmission and epidemiological characteristics of fascioliasis in the way for its control in other high altitude Andean endemic areas.

Impact of fascioliasis reinfection on Fasciola hepatica egg shedding: relationship with the immune-regulatory response

Fascioliasis is a disease caused by liver flukes. In human fascioliasis hyperendemic areas, reinfection and chronicity are the norm. Control strategies in humans require the use of egg count techniques to calculate the appropriate treatment dose for colic risk prevention. The present study investigates how fascioliasis reinfection affects liver fluke egg shedding and its relationship with the immune-regulatory response. The experimental design reproduced the usual reinfection/chronicity conditions in human fascioliasis endemic areas and included Fasciola hepatica primo-infected Wistar rats (PI) and rats reinfected at 4 weeks (R4), 8 weeks (R8), 12 weeks (R12), and negative control rats. In a longitudinal study (0-20 weeks post-infection, p.i.), serical IgG1 levels and eggs per gram of faeces (epg) were analyzed. In a cross-sectional study, the expression of the genes associated with Th1 (Ifng, Il12a, Il12b, Nos2), Th2 (Il4, Arg1), Treg (Foxp3, Il10, Tgfb, Ebi3), and Th17 (Il17) in the spleen and thymus was analyzed. In R8 and R12, transiently higher averages of epg and epg/worm in reinfected groups vs PI group were detected at least in the weeks following reinfection. The kinetics of IgG1 levels shows that reinfected groups followed a pattern similar to the one in the PI group, but transiently higher averages of IgG1 levels in reinfected groups vs the PI group were detected in the weeks following reinfection. Epg correlated with IgG1 levels and also with systemic Il10 and thymic Ifng, and Il10 expression levels. These results suggest that epg depends on the Th1 and Treg phenotype and that the determination of the fluke burden by epg is likely to be an overestimation in cases of recent reinfection in low burden situations. A strategy to facilitate the implementation of epg count techniques and the subsequent decision on the appropriate treatment dose for each patient to prevent colic risk is required.

Clinical Features, Diagnosis and Management of Patients with Suspicion of Fascioliasis in Kohgiluyeh and Boyer-Ahmad Province, Southwestern Iran

In the current study, we described the epidemiological features, clinical presentation, diagnosis and management of patients with suspicion of fascioliasis in Kohgiluyeh and Boyer-Ahmad Province in southwest of Iran. Overall, 56 patients with suspicion of fascioliasis, based on their clinical signs and symptoms that referred to Clinic of Internal Medicine in Yasuj city, from 2014 to 2016 were enrolled. Demographic data, history of eating aquatic local plants, the chief complains, and laboratory findings were recorded for each patient. Stool samples were obtained from each case for detection of Fasciola eggs. Moreover, blood samples were taken from each patient and evaluated for detection of anti-Fasciola antibodies by an indirect ELISA. Patients who defined as having fascioliasis were treated with triclabendazole and were followed for at least three months for clinical improvement. Serological test was positive in 5 patients. Of these 5 cases, three cases had a history of ingesting raw aquatic vegetables. The main clinical signs and symptoms in positive cases were; abdominal pain (60%), epigastric pain (40%), anemia (60%), and dermal pruritus (20%). Hypereosinophilia was seen in all of 5 positive cases. No Fasciola egg was found in stool specimens of any of the patients. The fascioliasis cases were treated by triclabendazole and clinical symptoms disappeared in all of 5 cases. Our observation further confirmed Yasuj district as a human endemic area for fascioliasis in Iran. The study also highlighted the importance of clinical features together with eosinophilia, as key parameters, in the diagnosis of human fascioliasis. Clinicians need to be aware of this disease and should keep in mind fascioliasis when hypereosinophilia present in patients in such endemic areas.

Equines as reservoirs of human fascioliasis: transmission capacity, epidemiology and pathogenicity in Fasciola hepatica-infected mules.

Fascioliasis is a zoonotic disease caused by liver flukes transmitted by freshwater lymnaeid snails. Donkey and horse reservoir roles have been highlighted in human endemic areas. Liver fluke infection in mules has received very limited research. Their role in disease transmission, epidemiological importance and Fasciola hepatica pathogenicity are studied for the first time. Prevalence was 39.5% in 81 mules from Aconcagua, and 24.4% in 127 from Uspallata, in high-altitude areas of Mendoza province, Argentina. A mean amount of 101,242 eggs/mule/day is estimated. Lymnaeids from Uspallata proved to belong to ribosomal DNA internal transcribed spacer (ITS) markers ITS-1 and ITS-2 combined haplotype 3C of Galba truncatula. These lymnaeids were experimentally susceptible to infection by egg miracidia from mules. Infectivity, number of cercariae/snail and shedding period fit the enhanced F. hepatica/G. truncatula transmission pattern at very high altitude. This indicates that the mule is able to maintain the F. hepatica cycle independently. Individual burdens of 20 and 97 flukes were found. Mule infection susceptibility is intermediate between donkey and horse, although closer to the latter. Anatomo-pathology and histopathology indicate that massive infection may cause mule death. Haematological value decreases of red blood cells, haemoglobin, leucocytes and lymphocytes indicate anaemia and strong immunosuppression. Strongly increased biochemical marker values indicate liver function alterations. The mule probably played a role in the past exchanges with Chile and Bolivia through Mendoza province. Evidence suggests that mules could contribute to the spread of both F. hepatica and G. truncatula to human fascioliasis-endemic areas in these countries.

Multiple microbiologic tests for tuberculosis improve diagnostic yield of bronchoscopy in medically complex patients

Background: Bronchoalveolar lavage (BAL) is indicated for medical evaluation of complex cases of lung disease. There is limited data on the performance of tuberculosis (TB) microbiologic tests on BAL in such patients, particularly in human immunodeficiency virus (HIV) and TB endemic areas. Methods: We evaluated the performance of Mycobacterium tuberculosis (Mtb) culture and up to two simultaneous Xpert MTB/RIF tests on BAL fluid against a consensus clinical diagnosis in 98 medically complex patients undergoing bronchoscopy over a two-year period in Durban, South Africa. Results: TB was the most frequently diagnosed lung disease, found in 19 of 98 participants (19%) and was microbiologically proven in 14 of these (74%); 9 (47%) were culture positive and 5 were positive on at least one Xpert MTB/RIF assay. Immunosuppression prevalence was high (26% HIV-infected, 29% on immunosuppressive therapy and 4% on chemotherapy). Xpert MTB/RIF had low sensitivity (45%) and high specificity (99%) when assessed against the consensus clinical diagnosis. Compared to TB culture, a single Xpert MTB/RIF increased the diagnostic yield by 11% and a second Xpert MTB/RIF by a further 16%. Conclusion: Although Xpert MTB/RIF had a low sensitivity, sending two tests improved the microbiologically-proven diagnostic yield of bronchoscopy from 47% to 74% compared to culture alone.

Multiple microbiologic tests for tuberculosis improve diagnostic yield of bronchoscopy in medically complex patients.

Background: Bronchoalveolar lavage (BAL) is indicated for medical evaluation of complex cases of lung disease. There is limited data on the performance of tuberculosis (TB) microbiologic tests on BAL in such patients, particularly in human immunodeficiency virus (HIV) and TB endemic areas. Methods: We evaluated the performance of Mycobacterium tuberculosis (Mtb) culture and up to two simultaneous Xpert MTB/RIF tests on BAL fluid against a consensus clinical diagnosis in 98 medically complex patients undergoing bronchoscopy over a two-year period in Durban, South Africa. Results: TB was the most frequently diagnosed lung disease, found in 19 of 98 participants (19%) and was microbiologically proven in 14 of these (74%); 9 (47%) were culture positive and 5 were positive on at least one Xpert MTB/RIF assay. Immunosuppression prevalence was high (26% HIV-infected, 29% on immunosuppressive therapy and 4% on chemotherapy). Xpert MTB/RIF had low sensitivity (45%) and high specificity (99%) when assessed against the consensus clinical diagnosis. Compared to TB culture, a single Xpert MTB/RIF increased the diagnostic yield by 11% and a second Xpert MTB/RIF by a further 16%. Conclusion: Although Xpert MTB/RIF had a low sensitivity, sending two tests improved the microbiologically-proven diagnostic yield of bronchoscopy from 47% to 74% compared to culture alone.

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

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

Vector and reservoir surveillance study in a canine and human leishmaniasis endemic area in most western part of Turkey, Karaburun

Leishmaniasis is an arthropod borne disease that is endemic in 102 countries and one and half million new cases are reported each year. Sand flies are the one and only proven vectors of the disease and dogs are the main reservoirs in urban areas. Karaburun peninsula is located in most western part of Turkey and is reported to be an endemic area for human and canine leishmaniasis. The most recent study was undertaken more than 15 years ago in The peninsula and no clear data available for vectors or reservoirs. Thus, we aimed to update the information regarding sand fly diversity, infection status of reservoirs and vectors in the study area.Sand flies were collected using CDC light traps at 13 different sites of Karaburun and species identification was made using previously published keys. Monospecific pools were generated using midguts with blood retention and were screened for the presence of Leishmania spp. DNA by molecular techniques. A non-invasive conjunctival swab sampling was performed to identify the infection status among reservoirs and species typing of the causative agent was also undertaken using ITS1 PCR.Three out of 30 pools were found positive for Leishmania infantum that were generated using guts of Phlebotomus tobbi (n:36). Among all sampled dogs (44) and cats (19), 11 and one of them were found positive for L. infantum, respectively. There was a decrease in the number of P. papatasi during the study period, while increase was observed in the number of P. tobbi.The presence of proven vectors and reservoirs as well as Leishmania DNA in cats was shown in the present study. Sand fly fauna is updated and Leishmania DNA presence in cats was reported in the study area for the first time.

First phenotypic and genotypic description of Fasciola hepatica infecting highland cattle in the state of Mexico, Mexico

Fascioliasis is a plantborne and zoonotic parasitic disease caused by fasciolid liver flukes. Fasciola hepatica is the only fasciolid species described in the Americas. Human fascioliasis endemic areas are mainly located in high altitude areas of the Americas. Given the necessity to characterize F. hepatica populations involved, the phenotypic and genotypic features of fasciolid adults infecting cattle in the highland area of Toluca, State of Mexico, Mexico, were analyzed and compared to fasciolid materials from the Northern Bolivian Altiplano, representing the altiplanic transmission pattern in a hyperendemic scenario. A computer image analysis system (CIAS) was applied on the basis of standardized measurements. The aforementioned F. hepatica highland populations were compared to standard lowland natural populations of European origin (Spain and France) and F. gigantica of African origin (Burkina Faso). Liver-fluke size was studied by principal component analysis (PCA). Two phenotypic patterns could be distinguished in the F. hepatica material analyzed from the Americas: the valley pattern (Toluca, Mexico) and the altiplanic pattern (Northern Altiplano, Bolivia). PCA showed that the Altiplano population presented a large body size range with a pronouncedly lower minimum size. Mahalanobis distances demonstrated that American populations are very close to European populations. Genetic haplotyping was performed using the ribosomal DNA intergenic region, including ITS-1, 5.8S and ITS-2. The intergenic region was 951 bp-long, providing 2 combined haplotypes due to one mutation appearing in the ITS-2 sequence. Molecular results showed that Fh-1A and Fh-2A, the most frequent haplotypes of F. hepatica from southern Europe, are present in Mexican cattle. Nuclear rDNA biomarkers correlated with adult fluke phenotypic characteristics. Results showed that the Mexican population analyzed and European standard populations presented a phenotypic and genotypic homogeneity, suggesting an introduction with livestock transported during the early colonization period. Results are moreover analyzed in terms of altitude and permanent/seasonal transmission characteristics.

Fasciola hepatica reinfection potentiates a mixed Th1/Th2/Th17/Treg response and correlates with the clinical phenotypes of anemia.

BackgroundFascioliasis is a severe zoonotic disease of worldwide extension caused by liver flukes. In human fascioliasis hyperendemic areas, reinfection and chronicity are the norm and anemia is the main sign. Herein, the profile of the Th1/Th2/Th17/Treg expression levels is analyzed after reinfection, correlating them with their corresponding hematological biomarkers of morbidity.Methodology/Principal findingsThe experimental design reproduces the usual reinfection/chronicity conditions in human fascioliasis endemic areas and included Fasciola hepatica primo-infected Wistar rats (PI) and rats reinfected at 8 weeks (R8), and at 12 weeks (R12), and negative control rats. In a cross-sectional study, the expression of the genes associated with Th1 (Ifng, Il12a, Il12b, Nos2), Th2 (Il4, Arg1), Treg (Foxp3, Il10, Tgfb, Ebi3), and Th17 (Il17) in the spleen and thymus was analyzed. After 20 weeks of primary infection, PI did not present significant changes in the expression of those genes when compared to non-infected rats (NI), but an increase of Il4, Arg1 and Ifng mRNA in the spleen was observed in R12, suggesting the existence of an active mixed Th1/Th2 systemic immune response in reinfection. Foxp3, Il10, Tgfb and Ebi3 levels increased in the spleen in R12 when compared to NI and PI, indicating that the Treg gene expression levels are potentiated in chronic phase reinfection. Il17 gene expression levels in R12 in the spleen increased when compared to NI, PI and R8. Gene expression levels of Il10 in the thymus increased when compared to NI and PI in R12. Ifng expression levels in the thymus increased in all reinfected rats, but not in PI. The clinical phenotype was determined by the fluke burden, the rat body weight and the hemogram. Multivariate mathematical models were built to describe the Th1/Th2/Th17/Treg expression levels and the clinical phenotype. In reinfection, two phenotypic patterns were detected: i) one which includes only increased splenic Ifng expression levels but no Treg expression, correlating with severe anemia; ii) another which includes increased splenic Ifng and Treg expression levels, correlating with a less severe anemia.Conclusions/SignificanceIn animals with established F. hepatica infection a huge increase in the immune response occurs, being a mixed Th2/Treg associated gene expression together with an expression of Ifng. Interestingly, a Th17 associated gene expression is also observed. Reinfection in the chronic phase is able to activate a mixed immune response (Th1/Th2/Th17/Treg) against F. hepatica but T and B proliferation to mitogens is strongly suppressed in all infected rats vs control in the advanced chronic phase independently of reinfection The systemic immune response is different in each group, suggesting that suppression is mediated by different mechanisms in each case. Immune suppression could be due to the parasite in PI and R8 rats and the induction of suppressive cells such as Treg in R12. This is the first study to provide fundamental insight into the immune profile in fascioliasis reinfection and its relation with the clinical phenotypes of anemia.

Human fascioliasis endemic areas in Argentina: multigene characterisation of the lymnaeid vectors and climatic-environmental assessment of the transmission pattern.

BackgroundIn South America, fascioliasis stands out due to the human endemic areas in many countries. In Argentina, human endemic areas have recently been detected. Lymnaeid vectors were studied in two human endemic localities of Catamarca province: Locality A beside Taton and Rio Grande villages; Locality B close to Recreo town.MethodsLymnaeids were characterised by the complete sequences of rDNA ITS-2 and ITS-1 and fragments of the mtDNA 16S and cox1. Shell morphometry was studied with the aid of a computer image analysis system. Climate analyses were made by nearest neighbour interpolation from FAO data. Koeppen & Budyko climate classifications were used. De Martonne aridity index and Gorczynski continentality index were obtained. Lymnaeid distribution was assessed in environmental studies.ResultsDNA sequences demonstrated the presence of Lymnaea neotropica and L. viator in Locality A and of L. neotropica in Locality B. Two and four new haplotypes were found in L. neotropica and L. viator, respectively. For interspecific differentiation, ITS-1 and 16S showed the highest and lowest resolution, respectively. For intraspecific analyses, cox1 was the best marker and ITS-1 the worst. Shell intraspecific variability overlapped in both species, except maximum length which was greater in L. viator. The desertic-arid conditions surrounding Locality A, the semiaridity-aridity surrounding Locality B, and the very low yearly precipitation in both localities, are very different from the typical fascioliasis transmission foci. Lymnaeids are confined to lateral river side floodings and small man-made irrigation systems. Water availability only depends on the rivers flowing from neighbouring mountains. All disease transmission factors are concentrated in small areas where humans and animals go for water supply, vegetable cultures and livestock farming.ConclusionsThe unusually high number of DNA haplotypes and the extreme climate unsuitable for F. hepatica and lymnaeid development, demonstrate that the transmission foci are isolated. Seasonal transmission may depend on the timely overlap of appropriate temperature and river water availability. Lymnaeids and F. hepatica have probably reached these localities by livestock introduction. DNA differences regarding other populations of L. neotropica and L. viator in Argentina suggest an introduction independent from the spreading movements which allowed these two lymnaeids to expand throughout the country.Electronic supplementary materialThe online version of this article (doi:10.1186/s13071-016-1589-z) contains supplementary material, which is available to authorized users.

Distribution of Fasciola hepatica and F. gigantica in the endemic area of Guilan, Iran: Relationships between zonal overlap and phenotypic traits

Fascioliasis is a zoonotic disease emerging in numerous parts of the world. In any endemic area, the characterisation of scenarios and patterns of infection must always be considered the starting point before implementing any control measure. Fascioliasis is a parasitic disease of different epidemiological, pathological and control characteristics depending on the endemic area and the causal agent, Fasciola hepatica and Fasciolagigantica. Classically it has been accepted that F. hepatica is present worldwide, while the distribution of the two species overlaps in many areas of Africa and Asia. Fascioliasis caused by F. hepatica, F. gigantica and intermediate forms is present in Guilan province, a complicated epidemiological situation where the highest human infection rates have been described in Iran. Morphometric tools were used to analyse the possible relationship between liver-fluke metric traits and geographical and altitudinal distribution. This is the first study in which a detailed distribution of both Fasciola species is analysed in a human fascioliasis endemic area with a zonal overlap transmission pattern. An accurate analysis was conducted to phenotypically discriminate between fasciolids from naturally infected livestock (cattle, buffaloes, sheep and goats). The distribution of the % F. hepatica-like (F.h.) and F. gigantica-like (F.g.) flukes detected in each liver versus altitude (m) in each group was analysed. The presence of F.g. specimens mainly in locations below sea level (average: 11.23% F.h., 88.77% F.g.), the presence of both species with similar intensity at 1–99m (average: 56.95% F.h., 43.05% F.g.) and the presence of F.h. specimens mainly from 100 to 999m (average: 71.69% F.h., 28.31% F.g.) as well as in locations with an altitude above 1000m (average: 97.48% F.h., 2.52% F.g.) are noteworthy. A significant positive correlation was obtained between altitude and % F.h., and a significant negative correlation was obtained between altitude and % F.g. The results show that F.g. populations in cattle, buffaloes and sheep share larger size values, but smaller specimens are present mainly in lowland populations located below sea level, independently of the host species (cattle, buffalo). F.g. from lowland cattle presented larger worm size variability. Four different fascioliasis transmission areas may be distinguished in Guilan: (a) lowland coastal areas neighbouring the Caspian Sea shore, below sea level, where basically F. gigantica-like specimens are found; (b) a coastal plain with an altitude between 1 and 100m where both species co-exist; (c) areas with altitude values of 100–999m where mainly F. hepatica-like specimens are found; (d) highland mountainous areas, where basically F. hepatica-like specimens are found. The study of the influence of the host species on the liver fluke was also carried out by a size-out analysis. This is the first report concerning the decisive influence exercised by the host species on the metric traits of F. gigantica adults.

Fascioliasis: a worldwide parasitic disease of importance in travel medicine.

Fascioliasis is a foodborne zoonotic disease caused by the two parasite species Fasciola hepatica and Fasciola gigantica. This trematodiasis has never been claimed special relevance for travellers and migrants. However, the situation has drastically changed in the last two decades, in a way that fascioliasis should today be included in the list of diseases to be enhanced in Travel Medicine. Different kind of travellers have been involved in human infection reports: business travellers, tourists, migrants, expatriated workers, military personnel, religious missionaries, and refugees. Europe is the continent where more imported cases have been reported in many countries. More cases would have been probably reported in Europe if fascioliasis would be a reportable disease. In the Americas, most of the reports concern cases diagnosed in USA. Relative few patients have been diagnosed in studies on travellers performed in Asia. In Africa, most cases were reported in Maghreb countries. Blood eosinophilia and the ingestion of watercress or any other suggestive freshwater plant in anamnesis are extremely useful in guiding towards a fascioliasis diagnosis in a developed country, although may not be so in human endemic areas of developing countries. Several suggestive clinical presentation aspects may be useful, although the clinical polymorphism may be misleading in many cases. Non-invasive techniques are helpful for the diagnosis, although images may lead to confusion. Laparoscopic visualization should assist and facilitate procurement of an accurately guided biopsy. Endoscopic retrograde cholangiopancreatography (ERCP) is the first choice in patients in the chronic phase. ERCP and sphincterotomy are used to extract parasites from the biliary tree. Fluke egg finding continues to be the gold standard and enables for burden quantification and establishing of the drug dose. Many serological and stool antigen detection tests have been developed. Immunological techniques present the advantages of being applicable during all periods of the disease, but fundamentally during the invasive or acute period, as well as to other situations in which coprological techniques may present problems. Triclabendazole is the drug of choice at present, although the spread of resistance to this drug is challenging. Prevention mainly concerns measures to avoid individual infection by considering the different human infection sources.

Diagnosis of human fascioliasis by stool and blood techniques: update for the present global scenario.

Before the 1990s, human fascioliasis diagnosis focused on individual patients in hospitals or health centres. Case reports were mainly from developed countries and usually concerned isolated human infection in animal endemic areas. From the mid-1990s onwards, due to the progressive description of human endemic areas and human infection reports in developing countries, but also new knowledge on clinical manifestations and pathology, new situations, hitherto neglected, entered in the global scenario. Human fascioliasis has proved to be pronouncedly more heterogeneous than previously thought, including different transmission patterns and epidemiological situations. Stool and blood techniques, the main tools for diagnosis in humans, have been improved for both patient and survey diagnosis. Present availabilities for human diagnosis are reviewed focusing on advantages and weaknesses, sample management, egg differentiation, qualitative and quantitative diagnosis, antibody and antigen detection, post-treatment monitoring and post-control surveillance. Main conclusions refer to the pronounced difficulties of diagnosing fascioliasis in humans given the different infection phases and parasite migration capacities, clinical heterogeneity, immunological complexity, different epidemiological situations and transmission patterns, the lack of a diagnostic technique covering all needs and situations, and the advisability for a combined use of different techniques, at least including a stool technique and a blood technique.

Fascioliasis and Intestinal Parasitoses Affecting Schoolchildren in Atlixco, Puebla State, Mexico: Epidemiology and Treatment with Nitazoxanide

BackgroundThe Atlixco municipality, Puebla State, at a mean altitude of 1840 m, was selected for a study of Fasciola hepatica infection in schoolchildren in Mexico. This area presents permanent water collections continuously receiving thaw water from Popocatepetl volcano (5426 m altitude) through the community supply channels, conforming an epidemiological scenario similar to those known in hyperendemic areas of Andean countries.Methodology and FindingsA total of 865 6–14 year-old schoolchildren were analyzed with FasciDIG coproantigen test and Lumbreras rapid sedimentation technique, and quantitatively assessed with Kato-Katz. Fascioliasis prevalences ranged 2.94–13.33% according to localities (mean 5.78%). Intensities were however low (24–384 epg). The association between fascioliasis and the habit of eating raw vegetables was identified, including watercress and radish with pronouncedly higher relative risk than lettuce, corncob, spinach, alfalfa juice, and broccoli. Many F. hepatica-infected children were coinfected by other parasites. Entamoeba histolytica/dispar, Giardia intestinalis, Blastocystis hominis, Hymenolepis nana and Ascaris lumbricoides infection resulted in risk factors for F. hepatica infection. Nitazoxanide efficacy against fascioliasis was 94.0% and 100% after first and second treatment courses, respectively. The few children, for whom a second treatment course was needed, were concomitantly infected by moderate ascariasis burdens. Its efficacy was also very high in the treatment of E. histolytica/E. dispar, G. intestinalis, B. hominis, H. nana, A. lumbricoides, Trichuris trichiura, and Enterobius vermicularis. A second treatment course was needed for all children affected by ancylostomatids.ConclusionsFascioliasis prevalences indicate this area to be mesoendemic, with isolated hyperendemic foci. This is the first time that a human fascioliasis endemic area is described in North America. Nitazoxanide appears as an appropriate alternative to triclabendazole, the present drug of choice for chronic fascioliasis. Its wide spectrum efficacy against intestinal protozooses and helminthiasis, usually coinfecting liver fluke infected subjects in human endemic areas, represents an important added value.

Molecular characterisation of Galba truncatula, Lymnaea neotropica and L. schirazensis from Cajamarca, Peru and their potential role in transmission of human and animal fascioliasis

BackgroundHuman and animal fascioliasis is emerging in many world regions, among which Andean countries constitute the largest regional hot spot and Peru the country presenting more human endemic areas. A survey was undertaken on the lymnaeid snails inhabiting the hyperendemic area of Cajamarca, where human prevalences are the highest known among the areas presenting a "valley transmission pattern", to establish which species are present, genetically characterise their populations by comparison with other human endemic areas, and discuss which ones have transmission capacity and their potential implications with human and animal infection.MethodsTherefore, ribosomal DNA ITS-2 and ITS-1, and mitochondrial DNA 16S and cox1 were sequenced by the dideoxy chain-termination method.ResultsResults indicate the presence of three, morphologically similar, small lymnaeid species belonging to the Galba/Fossaria group: Galba truncatula, Lymnaea neotropica and L. schirazensis. Only one combined haplotype for each species was found. The ITS-1, 16S and cox1 haplotypes of G. truncatula are new. No new haplotypes were found in the other two species. This scenario changes previous knowledge, in which only L. viator (= L. viatrix) was mentioned. Galba truncatula appears to be the most abundant, with high population densities and evident anthropophyly including usual presence in human neighbourhood. Infection by Fasciola hepatica larval stages were molecularly confirmed in two populations of this species. The nearness between G. truncatula populations presenting liver fluke infection and both human settings and schools for children, together with the absence of populations of other lymnaeid species in the locality, suggest a direct relationship with human infection.ConclusionsThe geographical overlap of three lymnaeid species poses problems for epidemiological studies and control action. First, a problem in classifying lymnaeid specimens in both field and laboratory activities, given their transmission capacity differences: G. truncatula mainly involved in transmission to humans, L neotropica typically responsible for livestock infection, and L. schirazensis unable for transmission. Although several phenotypic characteristics may be helpful for a preliminary specimen classification, a definitive classification can only be obtained by marker sequencing. Aditionally, L. schirazensis increases the confusion, owing to its ability to mix with other Galba/Fossaria species and distort fascioliasis data such as transmission capacity and infection susceptibility. Second, a problem for epidemiological analysis, surveillance and control by methods as mathematical modelling and Remote Sensing - Geographical Information Systems. In Cajamarca, low resolution mapping may be insufficient, as already verified in Andean areas where different lymnaeid species overlap.