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The miracidia hatching device. 1: lateral schematic drawing of the hatching flask; 2: three-dimensional schematic drawing of the exposing wood box; 3: the hatching flask inside the exposing box. Note the black rubber ring in the communicating duct; 4: detail of the upper part of the flask, showing the funnel; 5: the transportation box.
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It is still imperative to develop a parasitological technique highly sensitive for diagnosing schistosomiasis in epidemiological and individual surveys. A simple and cheap hatching device with a collecting container was manufactured and tested under experimental conditions. Twelve Kato-Katz slides were performed as golden standard for comparison. Q...
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Citations
... 29 The miracidium hatching test which depends on the positive phototrophic behavior of schistosome miracidia, has also demonstrated a valuable role in the detection process 30 where improvement of sensitivity of the test prevails. 31 On the other hand, the general shortcomings linked with the direct observation of ova by microscopy is normally inevitable. It is also remarkable that such adjustments are tedious and are often not selected for routine or extensive screening. ...
Schistosomiasis is the second most devastating parasite prevalent in the tropical region of the world, posing significant public health impacts in endemic areas. Presently, several disease mitigation measures have shown a decline in transmission of the infection rate in risk localities using mass drug administration (MDA) of school-based or community-wide treatments. Despite all the endeavors made, the decline in transmission of infection rate has not been attained in the entire medicated segment of the population. Perhaps the current challenges of control of the disease appear to be strongly associated with a lack of appropriate diagnostic tools. It’s well known that the current diagnosis of schistosomiasis greatly relies on conventional methods. On the other hand, minor symptoms of schistosomiasis and low sensitivity and specificity of diagnostic methods are still unresolved diagnostic challenges to clinicians. Numerous scholars have reviewed various diagnostic methods of schistosomiasis and attempted to identify their strengths and weaknesses, currently on function. As a result of the known limitations of the existing diagnostic tools, the need to develop new and feasible diagnostic methods and diagnostic markers is unquestionable for more precise detection of the infection. Hence, advances in diagnostic methods have been considered part of the solution for the control and eventual elimination strategy of the disease in endemic areas. As of today, easy, cheap, and accurate diagnostics for schistosomiasis are difficult to get, and this limits the concerted efforts towards full control of schistosomiasis. While looking for new diagnostic methods and markers, it is important to simultaneously work on improving the existing diagnostic methods for better results. This review tries to give new insights to the status of the existing diagnostic methods of schistosomiasis from conventional to modern via summarizing the strengths and limitations of the methods. It also tries to recommend new, sensitive and feasible diagnostic methods for future approaches.
... A previously reported miracidium hatching technique (MHT), which has been applied for diagnosis, requires specific equipment, artificial light, and is time consuming [4]. Thus, in the present study, the previous MHT protocol was modified and simplified to recover miracidia under field study for preparation of single-genome DNA that can be used for a population genetics study with microsatellite markers. ...
... To evaluate the promotive effect of light from several different sources in the miracidium hatching, the eggs were exposed at RT to room light, sunlight, fluorescent light and halogen light. For the room light condition, the plastic plate was kept on a bench in the laboratory [4,11,14]. For the sunlight condition, the plastic plate was kept beside a window inside a room from 10 in the morning in June. ...
In this study, a simple and efficient miracidium hatching technique (MHT) protocol for preparing a single-genome DNA of Schistosoma japonicum was proposed. The protocol was designed with 96-well plates to collect a miracidium for single-genome DNA preparation, and the effects of lighting conditions on hatching rates were evaluated. The highest hatching rate was recorded under sunlight (92.4%), followed by fluorescent light (88.0%), and the lowest rate was recorded under the dark condition (4.7%). The results suggested for the first time, to our knowledge, that sunlight was efficient for this simple MHT protocol. Successful amplification of microsatellite marker genes using DNA isolated from a single miracidium also confirmed the quality of the single-genome DNA for subsequent applications.
... When the presence of schistosome antibodies is determined, aetiological tests are required to confirm the diagnosis. The aetiological detection of schistosomes is based on egg detection 23 and the detection of miracidia after stool hatching 24 . Due to the large amount of faeces produced by livestock and the presence of higher levels of fibre in the faeces, egg detection is not a suitable option. ...
Currently the diagnosis of schistosomiasis is mainly determined by observing the presence of eggs in host stool samples. Because of the overwhelming number of impurities in the stool, eggs are rarely observed. Therefore, the stool hatching method is used to observe the miracidia in the water. However, the miracidia of Schistosoma japonicum are small and difficult to detect, and missed detection is likely to occur when the infection level is low. In this study, recombinant streptococcal protein G-enhanced green fluorescent protein (rSPG-EGFP) was expressed, purified, and used as a fluorescence staining reagent for miracidia. A preliminary miracidium fluorescence staining method based on antigen and antibody bindingwas established by combining recombinant protein staining with the stool hatching method. The stool hatching method was used to collect the miracidia of S. japonicum, and Schistosoma-positive serum and the recombinant protein were mixed to assess the feasibility of fluorescence staining of miracidia. The miracidia of S. japonicum and Schistosoma turkestanicum were incubated with S. japonicum-positive serum and S. turkestanicum-positive serum, respectively, to identify miracidia species. When the fluorescence staining method was used to observe living miracidia, the miracidiawere labelled by the recombinant protein, and their motility status was not affected.
... In the current study, the authors used miracidia hatching assay as the comparison as well as the gold-standard method for the detection of active schistosomiasis infection [20]. The application of hatching assay in the detection of S. japonicum infection from negative eggs is primarily based on miracidia behavior, the strong positive phototropic [59]. The observation of the moving hatched miracidia can be seen. ...
Background and Aim: Schistosomiasis japonica, a disease caused by Schistosoma japonicum, is a public health problem in the Philippines, the Republic of Indonesia, and the People's Republic of China. The disease is known as zoonotic, meaning other than humans, animals are involved as the reservoirs. In Indonesia, schistosomiasis surveillance in animals is not continuous. Thus, the study to determine the prevalence of the disease in animals is needed. The study was aimed to determine the seroprevalence of S. japonicum infection among four species of domestic animals in the Lindu Sub-district, Central Sulawesi Province of Indonesia.
Materials and Methods: Blood samples of domestic animals were collected and analyzed for the presence of anti-S. japonicum immunoglobulin G antibodies against S. japonicum soluble egg antigens using the indirect hemagglutination assay. Animal stool samples were collected, and the miracidia-hatching assay was used for the detection of S. japonicum infection. Additional data concerning the animal identity and the management practices were obtained through a questionnaire used in surveys and interviews.
Results: A total of 146 sera from 13 cattle, 24 buffaloes, 54 pigs, and 55 dogs were collected. The overall schistosomiasis seroprevalence was 64.4%. The serology prevalence in cattle, buffalo, pig, and dog was 100.0%, 41.7%, 74.1%, and 56.4%, respectively. Domestic animals in all of five villages have previous exposure with S. japonicum as seropositive animals detected in every village. A total of 104 animal stool samples from 146 animals sampled were obtained. The overall schistosomiasis prevalence determined by the miracidia hatching assay was 16.35%. The sensitivity and specificity of indirect hemagglutination assay (IHA) in the current study were 88.24% and 41.37%, respectively, with miracidia hatching assay as the gold-standard method.
Conclusion: This study has shown a high seroprevalence of schistosomiasis japonica among domestic animals in the Lindu Subdistrict. IHA can be used as the screening method for the detection of S. japonicum infection in domestic animals. Chemotherapy and animal livestock grazing management programs to reduce the parasite burden and Schistosoma egg contamination in the environment must be implemented as part of one health approaches, in addition to other control measures.
... This method is still recommended by the WHO for diagnosis at the community level. MHT can be done using the miracidia hatching device [50] or by sieving stool sample through a nylon tissue bag for concentrating the eggs. Then, hatching is carried out in a well-lit room at a temperature of 25 ± 3ºC followed by counting of the swimming miracidia [34]. ...
Caption: Schistosoma mansoni life cycle (1) Paired adult worms (female is thin, male is bigger), (2) Eggs (see note below), (3) Ciliated miracidium, (4) Biomphalaria intermediate host snail, (5) Sporocyst, (6) Cercariae, the infective stage, and (7) Schistosomula, the young parasite. Abstract Schistosomiasis is a neglected invasive worm disease with a huge disease burden in developing countries, particularly in children, and is seen increasingly in non-endemic regions through transfer by travellers, expatriates, and refugees. Undetected and untreated infections may be responsible for the persistence of transmission. Rapid and accurate diagnosis is the key to treatment and control. So far, parasitological detection methods remain the cornerstone of Schistosoma infection diagnosis in endemic regions, but conventional tests have limited sensitivity, in particular in low-grade infection. Recent advances contribute to improved detection in clinical and field settings. The recent progress in micro-and nanotechnologies opens a road by enabling the design of new miniaturized point-of-care devices and analytical platforms, which can be used for the rapid detection of these infections. This review starts with an overview of currently available laboratory tests and their performance and then discusses emerging rapid and micro/nanotechnologies-based tools. The epidemiological and clinical setting of testing is then discussed as an important determinant for the selection of the best analytical strategy in patients suspected to suffer from Schistosoma infection. Finally, it discusses the potential role of advanced technologies in the setting near to disease eradication is examined.
... In the context of low endemicity like ours, we strongly recommend the hatching method which guarantees a better diagnosis and offers the possibility of responding to many questions, in combination with the HRM in the context of the epidemiology of schistosomiasis in Gabon. The only inconvenience of these techniques is that they are very expensive and difficult to perform on a large scale or under field conditions [23]. DNA extraction is a key procedure, but it can be a methodological bottleneck in molecular diagnostic assays since preservation and initial quantity of stool to be examined directly affects the yield and quality of genomic DNA and thus the PCR results. ...
... This method is simple, and its potential for high sensitivity has been recognized but not standardized for quantitative measurement (Ross et al. 2001). More recently, a similar technique has been proposed for diagnosing schistosomiasis in individual or large-scale investigations (Jurberg et al. 2008). ...
Digenetic trematodes form a major group of human parasites, affecting a large number of humans, especially in endemic foci. Over 100 species have been reported infecting humans, including blood, lung, liver, and intestinal parasites. Traditionally, trematode infections have been diagnosed by parasitological methods based on the detection and the identification of eggs in different clinical samples. However, this is complicated due to the morphological similarity between eggs of different trematode species and other factors such as lack of sensitivity or ectopic locations of the parasites. Moreover, the problem is currently aggravated by migratory flows, international travel, international trade of foods, and changes in alimentary habits. Although efforts have been made for the development of immunological and molecular techniques, the detection of eggs through parasitological techniques remains as the gold standard for the diagnosis of trematodiases. In this chapter, we review the current status of knowledge on diagnostic techniques used when examining feces, urine, and sputum and also analyze the most relevant characteristics used to identify eggs with a quick key for the identification of eggs.
... A miracidia hatching apparatus was developed based on the potent positive phototropic behaviour of the miracidia [14]. Although the test is simple, cheap, useful for diagnosis and improving the diagnostic sensitivity [14], hatching can be affected by temperature and quality of water and is not suitable for routine, large-scale screening or in people with light infections [15]. ...
... A miracidia hatching apparatus was developed based on the potent positive phototropic behaviour of the miracidia [14]. Although the test is simple, cheap, useful for diagnosis and improving the diagnostic sensitivity [14], hatching can be affected by temperature and quality of water and is not suitable for routine, large-scale screening or in people with light infections [15]. ...
Schistosomiasis is a neglected tropical disease that affects about 290 million patients
worldwide. Children aged between 5 and 14 years represent 45.8% of the affected patients,
in addition, schistosomiasis has been reported in Schistosoma-free areas, mostly because
of tourism and immigration from endemic countries. Intestinal schistosomiasis caused by
Schistosoma mansoni is mainly diagnosed via direct stool examination for egg detection.
Immunological methods are favoured for disease monitoring and preliminary checking for
communities in areas with low infection rates, and for patients with light and chronic infections
where parasitological tests are negative. PCR-based diagnostic techniques are more sensitive,
but expensive. Tegument proteins and miRNAs are promising markers for diagnosis of
schistosomiasis. Here we review the diagnostic methods for schistosomiasis mansoni aiming
to reach a standardized technique for diagnosis of early infection to help better control of the
disease.
... CopE methods rely on direct detection and visualization of parasite eggs in feces and include the Kato-Katz (KK) thick smear procedure, which is a mainstay of control programs due to the relative ease of performing the test at low cost, although it lacks sensitivity in low-intensity infections [46,47] and FLOTAC [48,49]. Other diagnostic approaches include formal-ethyl acetate sedimentation-digestion (FEA-SD) [50], the Danish Bilharziasis Laboratory (DBL) technique [51,52], and the miracidial hatching technique (MHT) [53,54], which also tests egg viability. Molecular diagnostics rely on detection of parasite DNA in clinical samples, often stool but also in urine, blood, and saliva, and include loop-mediated isothermal amplification (LAMP) [55][56][57][58], conventional polymerase chain reaction (cPCR) [59,60], real-time PCR (qPCR) [28,[61][62][63][64], and digital droplet PCR (ddPCR) [65][66][67]. ...
Schistosomiasis is an infectious disease caused by helminth parasites of the genus Schistosoma. Worldwide, an estimated 250 million people are infected with these parasites with the majority of cases occurring in sub-Saharan Africa. Within Asia, three species of Schistosoma cause disease. Schistosoma japonicum is the most prevalent, followed by S. mekongi and S. malayensis. All three species are zoonotic, which causes concern for their control, as successful elimination not only requires management of the human definitive host, but also the animal reservoir hosts. With regard to Asian schistosomiasis, most of the published research has focused on S. japonicum with comparatively little attention paid to S. mekongi and even less focus on S. malayensis. In this review, we examine the three Asian schistosomes and their current status in their endemic countries: Cambodia, Lao People’s Democratic Republic, Myanmar, and Thailand (S. mekongi); Malaysia (S. malayensis); and Indonesia, People’s Republic of China, and the Philippines (S. japonicum). Prospects for control that could potentially lead to elimination are highlighted as these can inform researchers and disease control managers in other schistosomiasis-endemic areas, particularly in Africa and the Americas.
... Because of decomposition, we have less chance to detect eggs in droppings than to find them directly in the intestinal contents. Since the larvae in the eggs are soon destroyed in the manure, we cannot efficiently detect the presence of schistosome eggs if we wait for the miracidia to hatch, although this procedure has been used for a long time to inspect human stool or animal excreta (Terner, 1962;Goff and Ronald, 1980;Malek, 1980;Makundi et al., 1998;Jurberg et al., 2008;Zhu et al., 2014). ...
Additional geographical distribution of the Central European populations of Schistosoma turkestanicum and the detectability of their eggs in droppings were investigated in red deer samples, because this rare species had previously been shown only in a single Hungarian habitat. Samples from visceral organs, intestinal contents, and droppings on the ground from 11 hunting areas of Hungary were investigated to find a new presence of this fluke. Close to the first site of detection in the Gemenc forest another habitat along the southern border of the country was found where the parasite lives in red deer. Therefore, it is possible that the worm also occurs in neighbouring Serbia or Croatia. Schistosoma turkestanicum causes a low-intensity infection in red deer and this host sheds low amounts of eggs, therefore the eggs are difficult to detect. Droppings were cleared by sedimentation, filtered by sieve screening and then the eggs were flotated using solutions with an increasing density of 1200 g/L, 1300 g/L, 1350 g/L, and 1400 g/L while they were being stained red with acid fuchsin. Eggs in fresh faeces can be most efficiently separated from plant fibres using a flotation solution of 1350 g/L density, but in some cases eggs in old dung can be detected using a solution of a specific gravity lower or higher than that. By combining the advantages of the three concentration processes, eggs of S. turkestanicum, which are more recognisable by the red stain, can be found in samples in which they are present at a density lower than 1/g.
