Low-intensity Infections Research Articles

Influences of habitat and arthropod density on parasitism in two co-occurring host taxa

Habitat attributes are known to influence infectious diseases such as those caused by parasites, but most studies have only considered single host and (or) parasite taxa, making it difficult to assess which features may be of general importance and to predict how alterations could affect disease dynamics. We examined infection with trematode (flatworm) parasites in two commonly co-occurring host taxa (larval amphibians and larval odonates (dragonflies and damselflies)) to investigate links with landscape-level features, including agricultural activity. We also assessed pond community composition with respect to the abundance and richness of aquatic arthropods known to prey upon tadpoles and (or) free-swimming trematode infectious stages. Larval amphibians from agricultural sites were most likely to be parasitized but had lower infection intensities, and infected hosts were positively associated with increasing distance to the nearest forest habitat, but negatively with road distance. The opposite was observed for larval odonate infection status; however, probability and intensity of parasitism in both host taxa was negatively associated with greater predatory arthropod abundance, consistent with the “dilution effect” of biodiversity on infectious diseases. Our approach demonstrates the importance of considering multiple host taxa when studying habitat links to diseases, and future studies incorporating even greater diversity will be needed.

Point of care diagnosis of multiple schistosome parasites: Species-specific DNA detection in urine by loop-mediated isothermal amplification (LAMP)

Schistosomes are easily transmitted and high chance of repeat infection, so if control strategies based on targeted mass drug administration (MDA) are to succeed it is essential to have a test that is sensitive, accurate and simple to use. It is known and regularly demonstrated that praziquantel does not always eliminate an infection so in spite of the successes of control programs a residual of the reservoir survives to re-infect snails. The issue of diagnostic sensitivity becomes more critical in the assessment of program effectiveness. While serology, such as antigen capture tests might improve sensitivity, it has been shown that the presence of species-specific DNA fragments will indicate, most effectively, the presence of active parasites. Polymerase chain reaction (PCR) can amplify and detect DNA from urine residue captured on Whatman No. 3 filter paper that is dried after filtration. Previously we have detected S. mansoni and S. haematobium parasite-specific small repeat DNA fragment from filtered urine on filter paper by PCR. In the current study, we assessed the efficacy of detection of 86 urine samples for either or both schistosome parasites by PCR and loop-mediated isothermal amplification (LAMP) that were collected from a low to moderate transmission area in Ghana. Two different DNA extraction methods, standard extraction kit and field usable LAMP-PURE kit were also evaluated by PCR and LAMP amplification. With S. haematobium LAMP amplification for both extractions showed similar sensitivity and specificity when compared with PCR amplification (100%) verified by gel electrophoresis. For S. mansoni sensitivity was highest for LAMP amplification (100%) for standard extraction than PCR and LAMP with LAMP-PURE (99% and 94%). The LAMP-PURE extraction produced false negatives, which require further investigation for this field usable extraction kit. Overall high positive and negative predictive values (90% − 100%) for both species demonstrated a highly robust approach. The LAMP approach is close to point of care use and equally sensitive and specific to detection of species-specific DNA by PCR. LAMP can be an effective means to detect low intensity infection due to its simplicity and minimal DNA extraction requirement. This will enhance the effectiveness of surveillance and MDA control programs of schistosomiasis.

Disease ecology of Hematodinium perezi in a high salinity estuary: investigating seasonal trends in environmental detection.

The blue crab Callinectes sapidus has seen a general decline in population levels. One factor influencing mortality is infections by Hematodinium perezi, a dinoflagellate parasite. A 2 yr study was conducted in 2014 and 2015 to monitor H. perezi DNA within the Maryland (USA) coastal bays, comparing seasonal cycles in the abundance of parasite DNA in environmental samples to parasite presence in host blue crabs. A late summer to early fall peak in H. perezi infections in blue crabs was observed, consistent with previous work. Infection intensities matched this trend, showing a slow progression of low intensity infections early in the year, with a peak in moderate and heavy infections occurring between July and September, for both years. It was hypothesized that the peak in water column occurrence would coincide with those months when infection intensities were highest in blue crabs. As the peaks in water column occurrence were in July 2014 and August-September 2015, this is consistent with sporulation being the primary contributor to environmental detection in summer months. An additional peak in environmental detection occurred in both years during the early spring months, the cause of which is currently unknown but may be related to infections in overwintering crabs or alternate hosts. Several new crustacean hosts were identified within this estuary, including grass shrimp Palaemonetes spp. and the sand shrimp Crangon septemspinosa, as well as the mud crab Dyspanopeus sayi. Improved knowledge of this disease system will allow for better management of this important fishery.

Age‐related parasite load and longevity patterns in the sedge warbler Acrocephalus schoenobaenus

We report the results from a nine‐year study on parasite infection in males of a small migrant passerine, the sedge warbler Acrocephalus schoenobaenus. Every year, for each male caught during territory establishment we estimated infection intensity of two lineages of Haemoproteus belopolskyi (SW1 and SW3), using molecular methods. We found a significant relationship between infection intensity and males’ longevity in both studied lineages. There was severe mortality of second‐year sedge warblers after their first breeding season, with lower parasite load in survivors. The lower infection intensity in older age classes was related to between‐individual change in both lineages, but was also a result of differences in infection intensity during the lifetime of individual males in the SW3. The relation between parasite load and longevity suggests that parasite load may be an age‐dependent factor influencing individual survival.

Branchial Pathomorphology of Southern Bluefin Tuna Thunnus maccoyii (Castelnau, 1872) Infected by Helminth and Copepodan Parasites

Three metazoan parasites, a monogenean Hexostoma thynni and two species of copepods Pseudocycnus appendiculatus and Euryphorus brachypterus are known to parasitize the gills of ranched southern bluefin tuna (SBT) and other tuna species. However, there is no detailed information describing the pathological response to infection by these parasites in this species. Wild southern bluefin tuna Thunnus maccoyii (approximately 3 years of age), captured and towed to a grow-out site in the waters immediately south of Port Lincoln, South Australia were subsequently sampled (n = 10) monthly from March until August 2004 during commercial harvest operations. Longitudinal sections of gill hemibranchs with attached parasites were excised and fixed for routine histology and immunohistochemistry. Reference samples were also collected from fish displaying no signs of parasitism or other grossly observable anomalies. Two morphologically distinct granulocytes were observed and putatively identified as eosinophils and mast cells. Pathology was localized to filaments upon and immediately adjacent to parasite attachment sites. Branchial cellular responses, adjunct to the attachment of H. thynni by its opisthaptoral clamps, included hyperplasia and inflammation resulting in structural remodeling of branchial tissues. Inflammatory infiltrates were often dominated by putative eosinophils and lymphocytes when parasitized by H. thynni and P. appendiculatus. Gill associated lymphoid tissue infiltrated the lamellar regions particularly in response to helminth infection. A variable response ranging from hemorrhage with minor hyperplasia or fibroplasia and eosinophilic inflammation to a barely discernible change was seen for gill sections harboring P. appendiculatus and E. brachypterus. The magnitude of the host response to attachment by the latter was congruent with attachment proximity and parasite load. On the basis of the host responses reported here and the low intensity of infection observed in other associated studies these gill ectoparasites are currently considered a low risk for wild and ranched adult SBT.

Estimating prevalence of avian haemosporidians in natural populations: a comparative study on screening protocols

BackgroundBirds harbour an astonishing diversity of haemosporidian parasites. Renewed interest in avian haemosporidians as a model system has placed a greater emphasis on the development of screening protocols to estimate parasite prevalence and diversity. Prevalence estimates are often based on the molecular or blood-smear microscopy techniques. However, variation in diagnostic sensitivity among screening methodologies represents a potential source of bias that may lead to erroneous inference in comparisons of prevalence across studies. Here, we analyzed a suite of blood samples for the presence of parasites using four diagnostic tools and compared method-specific estimates of detection probability to assess the relative performance of screening strategies.MethodsWe screened a total of 394 bird blood samples collected in India (n = 203) and Sweden (n = 191) for the combined presence of Plasmodium, Haemoproteus and Leucocytozoon with three PCR assays: (i) qPCR; (ii) restriction enzyme-based assay; and (iii) nested protocol. In addition, we examined blood smears for estimates of parasite intensity which was further screened using qPCR method to evaluate if parasite intensity shows a relationship with qPCR (Ct values). Furthermore, we used single infected samples from parasite intensities: low, medium, high, very high to establish the reproducibility in qPCR.ResultsFor the combined data sets from India and Sweden, detection probability for submicroscopic and low intensity infections was highest for the qPCR method, followed by the nested protocol and the restriction enzyme-based assay. For high parasite intensities, the qPCR had high PCR reproducibility, with three out of three PCR replicates being positive and with consistent Ct values across all tenfold dilution series. For parasite intensities at very low and submicroscopic samples, the qPCR was reproducible in one out of the three replicates. The intensity of parasitemia estimated from smears showed inverse relationship with Ct values in both the Indian and Swedish data sets.ConclusionsOur study highlights the importance of accounting for methodological issues to better estimate infection in parasitological studies and illustrates how a wider deployment of diagnostic tools combined with statistical approaches is needed for each study, in order to provide adequate insight into the most appropriate approach to avoid erroneous inferences.

Parasites of the Lessepsian invasive fish Lagocephalus sceleratus (Gmelin 1789) in the eastern Mediterranean Sea

ABSTRACTLagocephalus sceleratus, is a Lessepsian invasive fish species introduced into the Mediterranean Sea through the Suez Canal. This study investigated it as host of parasites. Out of 41 individuals caught from northern and southern locations of the eastern Aegean Sea, nine were found to host parasites. Gnathia sp. praniza larva, an ectoparasite, was associated with severe damage of the primary and secondary structures of the gills and it was found in one specimen (prevalence 2.4%). Regarding endoparasites, Anisakis type I third-stage larvae were found in two specimens (prevalence 4.9%). In addition, adult nematodes of Hysterothylacium aduncum were found in another six specimens (prevalence 14.6%). Lagocephalus sceleratus is a new host for some indigenous parasites, with low host specificity, in the Mediterranean Sea, providing an additional niche for the success and increase of local populations of these parasites and, to our knowledge, this is the first record of L. sceleratus parasites in this area. Whether these parasites can control the invasion success of L. sceleratus needs to be further investigated. However, the low mean intensity of infection found in this study does not support such control.

Diagnostic Tests to Support Late-Stage Control Programs for Schistosomiasis and Soil-Transmitted Helminthiases.

Global efforts to address schistosomiasis and soil-transmitted helminthiases (STH) include deworming programs for school-aged children that are made possible by large-scale drug donations. Decisions on these mass drug administration (MDA) programs currently rely on microscopic examination of clinical specimens to determine the presence of parasite eggs. However, microscopy-based methods are not sensitive to the low-intensity infections that characterize populations that have undergone MDA. Thus, there has been increasing recognition within the schistosomiasis and STH communities of the need for improved diagnostic tools to support late-stage control program decisions, such as when to stop or reduce MDA. Failure to adequately address the need for new diagnostics could jeopardize achievement of the 2020 London Declaration goals. In this report, we assess diagnostic needs and landscape potential solutions and determine appropriate strategies to improve diagnostic testing to support control and elimination programs. Based upon literature reviews and previous input from experts in the schistosomiasis and STH communities, we prioritized two diagnostic use cases for further exploration: to inform MDA-stopping decisions and post-MDA surveillance. To this end, PATH has refined target product profiles (TPPs) for schistosomiasis and STH diagnostics that are applicable to these use cases. We evaluated the limitations of current diagnostic methods with regards to these use cases and identified candidate biomarkers and diagnostics with potential application as new tools. Based on this analysis, there is a need to develop antigen-detecting rapid diagnostic tests (RDTs) with simplified, field-deployable sample preparation for schistosomiasis. Additionally, there is a need for diagnostic tests that are more sensitive than the current methods for STH, which may include either a field-deployable molecular test or a simple, low-cost, rapid antigen-detecting test.

Study design and baseline results of an open-label cluster randomized community-intervention trial to assess the effectiveness of a modified mass deworming program in reducing hookworm infection in a tribal population in southern India.

IntroductionHookworm infection is a leading cause of iron deficiency anemia and malnutrition in resource-poor settings. Periodic mass deworming with anthelminthic drugs remains the cornerstone of hookworm control efforts worldwide. Reinfection following treatment occurs, reflecting the human host's inability to acquire immunity following exposure to an untreated reservoir of infection. This cluster randomized trial will evaluate the effectiveness of a modified, population-based, mass deworming strategy in reducing hookworm infection in an endemic southern Indian population.MethodsForty five tribal villages were randomized into three groups: one received annual treatment; the second received two rounds of treatment at 1-month intervals; and the third received four rounds of treatment – two rounds 1 month apart at the beginning, followed by another two after 6 months. Stool samples collected through cross-sectional parasitological surveys pre- and post-intervention, and at 3-monthly intervals for a period of 1 year were tested for presence of hookworm ova. Long-term effectiveness of treatment will be assessed through another survey conducted 2 years after the last treatment cycle.ResultsFrom a population of 11,857 individuals, 8681 (73.2%) were found to be eligible and consented to participate, out-migration being the primary reason for non-participation. Baseline stool samples were obtained from 2082 participants, with 18.5% having hookworm infection, although majority were low intensity infections (<2000 eggs per gram of feces).DiscussionThis study will help identify the optimal mass deworming strategy that can achieve the greatest impact in the shortest period of time, particularly in settings where long-term program sustainability is a challenge.

Trematode fauna (Platyhelminthes: Digenea) of some sea ducks wintering on the southern Baltic coast (NW Poland) – a general comparison

ABSTRACTThe common scoter Melanitta nigra, velvet scoter M. fusca and long-tailed duck Clangula hyemalis (Aves: Anatidae) are among the more abundant sea ducks on the southern coast of the Baltic in winter, but their parasitic trematode fauna (Platyhelminthes, Digenea) in this region is little known. The aim of this study, conducted in 2007–2009, was a general comparison of the trematode fauna of these ducks in NW Poland. The prevalence and intensity of trematode infection were 68.3% and 119.9 in common scoter (101 examined), 77.6% and 13.1 in velvet scoter (58 examined) and 70.6% and 23.7 in long-tailed duck (68 examined). The following trematodes were found: Diplostomum pungitii, Apatemon gracilis, Australapatemon minor, Typhlocoelum sisowi, Echinochasmus spinulosus, Echinoparyphium recurvatum, Psilostomum brevicolle, Psilotrema simillimum, Catatropis verrucosa, Notocotylus attenuatus, N. gibbus, Paramonostomum alveatum, Prosthogonimus ovatus, Levinseniella propinqua, Gymnophallus bursicola and Cryptocotyle concava. The common scoter differs from the velvet scoter and long-tailed duck in its statistically significantly higher prevalence of G. bursicola; from long-tailed duck in its higher prevalence and greater intensity of infection with P. alveatum; and from velvet scoter in its greater intensity of infection with C. concava. The velvet scoter is distinguished from the long-tailed duck by its greater intensity of infection with P. brevicolle, lower prevalence of L. propinqua and lower intensity of infection with C. concava. Links between the differences found and the biology and ecology of the hosts, as well as the means of transmission of the parasites, are also discussed.

COMPARISON OF CURRENT METHODS FOR THE DETECTION OF CHRONIC MYCOPLASMAL URTD IN WILD POPULATIONS OF THE MOJAVE DESERT TORTOISE (GOPHERUS AGASSIZII).

Pathogens that cause subclinical diseases or exhibit low infection intensities are difficult to quantify in wild populations. Mojave desert tortoises ( Gopherus agassizii ) have been the focus of much research aimed at measuring the presence of upper respiratory disease (URTD) and URTD-associated pathogens, and techniques used to quantify disease in Gopherus species have also been used for disease surveillance in other species of turtles and tortoises of conservation concern. Published surveys of G. agassizii populations have found a relatively low prevalence of URTD, with most URTD-positive animals exhibiting moderate, intermittent signs of morbidity. Therefore, multiple tests have been developed to quantify URTD including genetic detection of the pathogens Mycoplasma agassizii and Mycoplasma testudineum , detection of M. agassizii -specific antibodies, and standardized quantification of clinical signs of URTD and body condition. These diagnostic tests have only been compared in diseased or moribund, semicaptive animals. We compared diagnostic techniques (TaqMan® and SYBR™ Green qPCR, serology, and visible examination) to detect M. agassizii -associated URTD in 126 wild desert tortoises sampled in Nevada and California, US in 2010. All had healthy body condition indices and none exhibited more than mild-to-moderate visual signs of URTD. Pairwise comparisons of diagnostic techniques indicated poor performance in diagnosing disease in individual animals. We found stronger, but inconsistent, statistical associations among diagnostic techniques at the population level. Our findings have implications for quantifying subclinical respiratory disease in tortoises.

Development and evaluation of a Markov model to predict changes in schistosomiasis prevalence in response to praziquantel treatment: a case study of Schistosoma mansoni in Uganda and Mali

BackgroundUnderstanding whether schistosomiasis control programmes are on course to control morbidity and potentially switch towards elimination interventions would benefit from user-friendly quantitative tools that facilitate analysis of progress and highlight areas not responding to treatment. This study aimed to develop and evaluate such a tool using large datasets collected during Schistosomiasis Control Initiative-supported control programmes.MethodsA discrete-time Markov model was developed using transition probability matrices parameterized with control programme longitudinal data on Schistosoma mansoni obtained from Uganda and Mali. Four matrix variants (A-D) were used to compare different data types for parameterization: A-C from Uganda and D from Mali. Matrix A used data at baseline and year 1 of the control programme; B used year 1 and year 2; C used baseline and year 1 from selected districts, and D used baseline and year 1 Mali data. Model predictions were tested against 3 subsets of the Uganda dataset: dataset 1, the full 4-year longitudinal cohort; dataset 2, from districts not used to parameterize matrix C; dataset 3, cross-sectional data, and dataset 4, from Mali as an independent dataset.ResultsThe model parameterized using matrices A, B and D predicted similar infection dynamics (overall and when stratified by infection intensity). Matrices A-D successfully predicted prevalence in each follow-up year for low and high intensity categories in dataset 1 followed by dataset 2. Matrices A, B and D yielded similar and close matches to dataset 1 with marginal discrepancies when comparing model outputs against datasets 2 and 3. Matrix C produced more variable results, correctly estimating fewer data points.ConclusionModel outputs closely matched observed values and were a useful predictor of the infection dynamics of S. mansoni when using longitudinal and cross-sectional data from Uganda. This also held when the model was tested with data from Mali. This was most apparent when modelling overall infection and in low and high infection intensity areas. Our results indicate the applicability of this Markov model approach as countries aim at reaching their control targets and potentially move towards the elimination of schistosomiasis.Electronic supplementary materialThe online version of this article (doi:10.1186/s13071-016-1824-7) contains supplementary material, which is available to authorized users.

Hyperparasitism by the bacteriophage (Caudovirales) infecting Candidatus Xenohaliotis californiensis (Rickettsiales-like prokaryote) parasite of wild abalone Haliotis fulgens and Haliotis corrugata from the Peninsula of Baja California, Mexico

Candidatus Xenohaliotis californiensis (CXc) is a Rickettsiales-like prokaryote that is considered the causal agent of Withering Syndrome (WS), a chronic disease of abalone, from the west coast of North America and it is listed by the International Organization for Animal Health (OIE) as a reportable agent due to its pathogenicity. This bacterium in red abalone Haliotis rufescens, black abalone Haliotis cracherodii, and yellow abalone Haliotis corrugata from California, US and Baja California, Mexico has been found to be infected by a bacteriophage. To date, there is no information on the epizootiology of CXc and its bacteriophage in natural populations of abalone; furthermore, it is unknown if the bacteriophage was also present in CXc infecting blue abalone Haliotis fulgens. The objective of this study was to determine the distribution, prevalence and intensity of CXc, as well as to determine the distribution and prevalence of the bacteriophage and to study interactions between host sex and hyperparasitism in blue abalone and yellow abalone. Tissue samples were obtained from seven localities where the commercial capture of wild abalone is carried out. Samplings were conducted throughout the 2012–2013 capture seasons and a total of 182 blue abalone and 170 yellow abalone were obtained. The prevalence and intensity of CXc and the prevalence of the bacteriophage were determined by histology. The identity of CXc was confirmed by PCR, product sequence analysis and in situ hybridization while the identity of the bacteriophage was corroborated by TEM. The prevalence of CXc infected and uninfected by the bacteriophage was 80% in blue abalone and 62% in yellow abalone. Low infection intensities were found in 86% of blue abalone and 82% of yellow abalone. Infection intensity was significantly higher in undifferentiated yellow abalone. The bacteriophage in CXc showed a prevalence of 22% and 31% in blue abalone and yellow abalone respectively. These results show that CXc and its bacteriophage are widely distributed in the peninsula of Baja California and that they are well established in natural populations of blue abalone and yellow abalone. Additionally, this data constitutes the first record of a bacteriophage in blue abalone.

Evaluation of field recognizable morbidity indicators of Schistosoma haematobium infection among primary school children in Ghana

Aim - This study was a retrospective evaluation of morbidity indicators and the use of questionnaire in decisions that relate to Schistosoma haematobium infection control among primary school children in Ghana. Materials and Methods — In all, 417 pupils in three primary schools (Kasseh sub-District, Dangme East District, Ghana), provided urine samples, which were examined for S. haematobium ova. The study utilized a structured questionnaire, the Urit 10V multi-stick urine reagent strips and the filtration method. Results — Overall, prevalence by microscopy was 20.9% (requires targeted treatment once every two years), and was higher (14.0%) among the 10-15 years age group (χ2=22.44, p<0.001). The lowest prevalence was among the <6 (0.5%, 2/417) and ≥19 (1.4%, 6/417) years age groups. There was no significant difference in prevalence among females (7.9%, 33/417) and males (12.9%, 54/417) (χ2=2.58, p=0.108). Self-reported haematuria was 43.2% (targeted treatment once a year). There was a significant difference between female (16.1%, 67/417) and male (27.1%, 113/417) responses to self-reported haematuria (χ2=4.170, p<0.001). A lower consolidated mean prevalence of 11.7% was estimated among pupils who had ova in their urine and also exhibited three morbidity indicators (haematuria, leucocyturia and proteinuria). Combined and averaged sensitivity and specificity estimations (sensitivity = 55.4%, specificity = 67.8%, Positive Predictive Value (PPV) = 53.4%, Negative Predictive Value (NPV) = 64.2%) of these morbidity indicators, was insufficient in equalizing the reliability of ova quantification. With the exception of leucocyturia, which was common among all pupils, 28.75% (25/87) of the infected individuals did not have any form of haematuria or proteinuria. Out of these, one pupil had heavy ova intensity (80 eggs/10ml of urine) while the remaining 24 had low ova intensity infection (1-44 eggs/10ml of urine). Questionnaire correctly identified 56.3% of pupils with S. haematobium ova in their urine, also reporting bloody urine (PPV=27.2%; NPV=84.0%). It also identified 49.4% of pupils with ova, reporting painful urination (PPV=26.1%, NPV=82.5%). Conclusion — This study emphasizes the relevance of evaluating the criteria for interpretation of morbidity indicators, prior to their field application. Questionnaire use remains key for surveillance purposes, to determine the presence of the infection in an area. Future studies should consolidate similar data elsewhere in Ghana for a better understanding of morbidity indicators and questionnaire use for decisions that relates to control strategies.

Larvae of Contracaecum sp. (Nematoda: Anisakidae) in the threatened freshwater fish Sandelia capensis (Anabantidae) in South Africa

Third-stage larvae of the nematode genus Contracaecum Railliet et Henry, 1912 (Contracaecum sp.) were, for the first time, recorded from the abdominal cavity of the threatened endemic freshwater fish Sandelia capensis (Cuvier) in South Africa. The larval morphology indicated that they belong to a species of which the adults are parasitic in fish-eating birds. Although the nematode seems to be a common parasite of S. capensis in the locality under study (prevalence 23%), the low intensity of infection recorded (1 to 4) and the generally known low pathogenicity of Contracaecum larvae in fish indicate that this parasite probably does not represent a danger to the local population of this threatened fish species.

Morbidity assessment of Opisthorchis viverrini infection in rural Laos: I. Parasitological, clinical, ultrasonographical and biochemical findings.

BackgroundInfections with the food-borne trematode Opisthorchis viverinni are common in Southeast Asia. In Lao PDR alone, two million people are supposed to be infected. Opisthorchiasis may cause severe liver disease, eventually leading to cholangiocarcinoma. The objective of this study is to assess the eating habits, complaints, symptoms, signs and ultrasonographical findings in three different areas of Savannakhet Province.MethodsStudy participants were recruited in Lahanam village in the flood-prone lowland of, Sonkhone district, Savannakhet Province (group A); in Non Somboon village, a community located on a hilly plateau in the same district (group B); and in staff of Savannakhet Province Hospital, Savannakhet town (group C). Eating habits, complaints and symptoms were recorded by standardized structured questionnaires. Participants were thoroughly examined clinically, and ultrasonography was performed. O. viverrini eggs were looked for in stool and in duodenal fluid. An array of biochemical and haematological parameters potentially related to liver disease was determined. Group A consisted of 45, group B of 31 and group C of 18 individuals.ResultsEating habits were similar in the three groups, except that participants from group C tended to consume less high-risk types of fish dishes and more frequently ate beef and pork. Average intensity of infection (eggs per gram of stool) was low, but significantly higher in group A than in group B and C (p < 0.001). Medical history and complaints were similar in the three groups. Ultrasonography did not reveal any bile duct pathology. The only pathological finding was a slight elevation of ASAT and gamma-GT in a few participants in groups A and B.ConclusionsThe study shows that eating habits favouring the infection with O. viverrini are common in south Laos. Although the average intensity of infection was low, there was a significant difference between the groups, paralleling slightly different eating habits. Clinically, this corresponded to a paucity of liver disease-associated complaints and signs. The low intensity of infection probably explains why no alterations of bile ducts were detectable by ultrasonography.

MHC‐I provides both quantitative resistance and susceptibility to blood parasites in blue tits in the wild

Major histocompatibility complex (MHC) genes are central for the adaptive immune response against parasites. Here, we investigated potential associations among MHC‐I alleles and blood parasite infections in a natural breeding population of a passerine bird, the blue titCyanistes caeruleus, in central Spain. We screened both infection status (presence/absence of infection) and infection intensity to the pathogenic blood parasitesHaemoproteusandLeucocytozoon. Three MHC‐I alleles (UA104, UA108 and UA117) were associated with higher or lower infection intensities byLeucocytozoon. Interestingly, these associations were dependent on age and were found both among young and adult birds. No MHC alleles were associated with infection intensity byHaemoproteusparasites. In addition, no significant relationships were detected between infection status byHaemoproteusandLeucocytozooninfections and MHC alleles. The very high prevalence of these two parasites in our study population (79–100%) poses challenges to identify associations with infection status and also suggests that clearance of infections may be rare. In conclusion, associations between specific MHC‐I alleles andLeucocytozoonparasites were related to either high or low infection intensities, and hence increased susceptibility or resistance to infection.

Optimisation of a droplet digital PCR assay for the diagnosis of Schistosoma japonicum infection: A duplex approach with DNA binding dye chemistry

Schistosomiasis is a chronically debilitating helminth infection with a significant socio-economic and public health impact. Accurate diagnostics play a pivotal role in achieving current schistosomiasis control and elimination goals. However, many of the current diagnostic procedures, which rely on detection of schistosome eggs, have major limitations including lack of accuracy and the inability to detect pre-patent infections. DNA-based detection methods provide a viable alternative to the current tests commonly used for schistosomiasis diagnosis. Here we describe the optimisation of a novel droplet digital PCR (ddPCR) duplex assay for the diagnosis of Schistosoma japonicum infection which provides improved detection sensitivity and specificity. The assay involves the amplification of two specific and abundant target gene sequences in S. japonicum; a retrotransposon (SjR2) and a portion of a mitochondrial gene (nad1). The assay detected target sequences in different sources of schistosome DNA isolated from adult worms, schistosomules and eggs, and exhibits a high level of specificity, thereby representing an ideal tool for the detection of low levels of parasite DNA in different clinical samples including parasite cell free DNA in the host circulation and other bodily fluids. Moreover, being quantitative, the assay can be used to determine parasite infection intensity and, could provide an important tool for the detection of low intensity infections in low prevalence schistosomiasis-endemic areas.

Proteomic Patterns in Serum and Urine of Patients Infected with Schistosoma mansoni: Basis for New Biomarker

Diagnosis of S. mansoni infection is carried out by the detection of eggs in faeces. This method has low sensitivity, especially with patients in the acute phase of the illness or with low-intensity infection. Serological tests cannot differentiate between active and past infection. Non-invasive diagnostic tests for active disease and monitoring response to treatment are required. This study was performed to identify serum and urine proteomic based biomarkers in patients with S. mansoni infection before and after therapeutic intervention using Surface Enhanced Laser Desorption/Ionization Time of Fight-Mass Spectrometry (SELDI TOF-MS). Serum samples were collected from 30 patients and urine samples from another15 patients both prior to and four to six weeks after treatment with praziquantel. All patients had a confirmed diagnosis of active infection on rectal biopsy. Serum and urine proteomic profiles were obtained by SELDI TOF-MS using cation capture (CM10) and immobilized metal affinity (IMAC30) ProteinChip ™ arrays. In urine samples, nine protein peaks demonstrated significant differences between pre and post treatment samples: 46 kDa, 44 kDa, 34 KDa, 13.3 KDa, 10.8 KDa, 19.7 kDa, 15.9 kDa, 18.1 kDa, 4.7 KDa (All had p<0.05). On ROC curve analysis, only the protein at 4.7 kDa showed a significant diagnostic signal (AUROC=0.77) indicating that it has potential as biomarker for active infection. In serum samples, only four peaks were found to be significantly different between pre and post treatment groups (p value<0.01). However no serum peak demonstrated significance on ROC curve analysis. These results suggest that urinary proteomic testing may provide a non-invasive diagnostic test for S. mansoni.

Development of a Loop Mediated Isothermal Amplification for Diagnosis of Ascaris lumbricoides in Fecal Samples.

Ascaris lumbricoides is a nematode parasite that causes the common tropical infection ascariasis in humans. It is also considered among the neglected tropical diseases. Diagnosis relies mainly on microscopy-based methods which are laborious, are limited by low sensitivity, and require high expertise. We have developed a loop mediated isothermal amplification (LAMP) for diagnosis of ascariasis in fecal samples, based on the first internal transcribed (ITS-1) spacer region of the ribosomal DNA. We used Primer Explorer V4 software to design primers. Ascaris adult and ova were obtained from naturally infected school children, whose parents/guardians gave consent for their participation in the study. Genomic DNA was extracted using alkaline lysis method and amplified by LAMP at 63°C for 45 minutes. LAMP products were visualized by naked eyes after adding SYBR Green dye and also on agarose gel. LAMP successfully and reliably detected Ascaris DNA from a single egg and in fecal samples. The assay specifically detected Ascaris DNA without amplifying DNA from ova of other parasites which commonly coexist with A. lumbricoides in feces. The developed LAMP assay has great potential for use in ascariasis diagnosis at the point of care and in low infection intensity situation that characterize control and elimination campaigns.