23
views
0
recommends
+1 Recommend
0 collections
    4
    shares
      • Record: found
      • Abstract: found
      • Article: found
      Is Open Access

      Molecular Epidemiology of Amoebiasis: A Cross-Sectional Study among North East Indian Population

      research-article

      Read this article at

      Bookmark
          There is no author summary for this article yet. Authors can add summaries to their articles on ScienceOpen to make them more accessible to a non-specialist audience.

          Abstract

          Background

          Epidemiological studies carried out using culture or microscopy in most of the amoebiasis endemic developing countries, yielded confusing results since none of these could differentiate the pathogenic Entamoeba histolytica from the non-pathogenic Entamoeba dispar and Entamoeba moshkovskii. The Northeastern part of India is a hot spot of infection since the climatic conditions are most conducive for the infection and so far no systemic study has been carried out in this region.

          Methodology/Principal Findings

          Following a cross-sectional study designed during the period 2011–2014, a total of 1260 fecal samples collected from the Northeast Indian population were subjected to microscopy, fecal culture and a sensitive and specific DNA dot blot screening assay developed in our laboratory targeting the Entamoeba spp. Further species discrimination using PCR assay performed in microscopy, culture and DNA dot blot screening positive samples showed E. histolytica an overall prevalence rate of 11.1%, 8.0% and 13.7% respectively. In addition, infection rates of nonpathogenic E. dispar and E. moshkovskii were 11.8% (95% CI = 10.2, 13.8) and 7.8% (95% CI = 6.4, 9.4) respectively. The spatial distributions of infection were 18.2% (107/588) of Assam, 11.7% (23/197) of Manipur, 10.2% (21/207) of Meghalaya, and 8.2% (22/268) of Tripura states. Association study of the disease with demographic features suggested poor living condition (OR = 3.21; 95% CI = 1.83, 5.63), previous history of infection in family member (OR = 3.18; 95% CI = 2.09, 4.82) and unhygienic toilet facility (OR = 1.79; 95% CI = 1.28, 2.49) as significant risk factors for amoebiasis. Children in age group <15 yr, participants having lower levels of education, and daily laborers exhibited a higher infection rate.

          Conclusions/Significance

          Despite the importance of molecular diagnosis of amoebiasis, molecular epidemiological data based on a large sample size from endemic countries are rarely reported in the literature. Improved and faster method of diagnosis employed here to dissect out the pathogenic from the nonpathogenic species would help the clinicians to prescribe the appropriate anti-amoebic drug.

          Author Summary

          Most epidemiologic studies in developing countries carried out for amoebiasis is either based on microscopy alone or culture/ microscopy used as a screening tool, have poor sensitivity and specificity and thus fails to figure out its true magnitude. The purpose of this study was to assess the true prevalence of amoebiasis in selected North Eastern states of India using DNA based screening technique followed by PCR assay for species discrimination. In addition, PCR assay confirmed that only 55.8% of the samples, resembling E. histolytica by microscopy, were true E. histolytica, implying that remaining 44.2% of so-called infections were due to other nonpathogenic Entamoeba spp. We found a higher prevalence of amebiasis (13.7%) using DNA dot blot screening compared to conventional microscopy and culture based screening. Poor living condition, previous history of infection in a family member, unhygienic toilet facility, children in age group <15 yr, participants having lower levels of education and daily laborers were identified as significant risk factors for amoebiasis. Thus, the techniques like DNA dot blot hybridization and PCR based detection adopted in the present study over and above the conventional screening methods can reduce misdiagnosis of the disease appreciably from the population living in this endemic area.

          Related collections

          Most cited references55

          • Record: found
          • Abstract: found
          • Article: not found

          Laboratory diagnostic techniques for Entamoeba species.

          The genus Entamoeba contains many species, six of which (Entamoeba histolytica, Entamoeba dispar, Entamoeba moshkovskii, Entamoeba polecki, Entamoeba coli, and Entamoeba hartmanni) reside in the human intestinal lumen. Entamoeba histolytica is the causative agent of amebiasis and is considered a leading parasitic cause of death worldwide in humans. Although recent studies highlight the recovery of E. dispar and E. moshkovskii from patients with gastrointestinal symptoms, there is still no convincing evidence of a causal link between the presence of these two species and the symptoms of the host. New approaches to the identification of E. histolytica are based on detection of E. histolytica-specific antigen and DNA in stool and other clinical samples. Several molecular diagnostic tests, including conventional and real-time PCR, have been developed for the detection and differentiation of E. histolytica, E. dispar, and E. moshkovskii in clinical samples. The purpose of this review is to discuss different methods that exist for the identification of E. histolytica, E. dispar, and E. moshkovskii which are available to the clinical diagnostic laboratory. To address the need for a specific diagnostic test for amebiasis, a substantial amount of work has been carried out over the last decade in different parts of the world. The molecular diagnostic tests are increasingly being used for both clinical and research purposes. In order to minimize undue treatment of individuals infected with other species of Entamoeba such as E. dispar and E. moshkovskii, efforts have been made for specific diagnosis of E. histolytica infection and not to treat based simply on the microscopic examination of Entamoeba species in the stool. The incorporation of many new technologies into the diagnostic laboratory will lead to a better understanding of the public health problem and measures to control the disease.
            Bookmark
            • Record: found
            • Abstract: found
            • Article: not found

            Prevalence and Risk Factors of Intestinal Parasitism in Rural and Remote West Malaysia

            Introduction Globally, the neglected intestinal parasitic infections (IPIs) such as soil-transmitted helminth (STH) and protozoa infections have been recognized as one of the most significant causes of illnesses and diseases especially among disadvantaged communities. With an average prevalence rate of 50% in developed world, and almost 95% in developing countries, it is estimated that IPIs result in 450 million illnesses [1], [2], [3]. These infections are ubiquitous with high prevalence among the poor and socioeconomically deprived communities where overcrowding, poor environmental sanitation, low level of education and lack of access to safe water are prevalent [4], trapping them in a perennial cycle of poverty and destitution [5]. These parasitic diseases contribute to economic instability and social marginalization; and the poor people of under developed nations experience a vicious cycle of under nutrition and repeated infections leading to excess morbidity with children being the worst affected [2], [6]. Of these illnesses, infections by STH have been increasingly recognized as an important public health problem and most prevalent of IPIs [7]. STH infections caused by Ascaris lumbricoides, Trichuris trichiura and hookworm (Necator americanus and Ancylostoma duodenale) are most significant in the bottom billion of the world's poorest people (i.e., RM 500 54668 45627 83.540.0 7.60 (5.30–11.13)1 <0.001 Water supply status*Untreated (river, well, rain water)Treated pipe water 317399 275249 86.862.4 2.84 (2.08–3.86)1 <0.001 Presence of proper latrine systemNoYes 212504 181343 85.468.1 2.19 (1.54–3.10)1 <0.001 Type of toilet facilityNonePour flush toilet 538178 44678 82.943.8 3.29 (2.62–4.12)1 <0.001 Defecation places status*Others (Bush, River)Pour flush toilet 550166 45668 82.941.0 3.45 (2.76–4.32)1 <0.001 Close contact with pets/livestockYesNo 65165 47252 72.580.0 0.73 (0.44–1.20)1 0.193 Garbage disposalIndiscriminatelyCollected 198518 168356 84.868.7 2.06 (1.45–2.94)1 <0.001 Iron supplementNoYes 412304 303221 73.572.7 1.03 (0.81–1.32)1 0.801 Anthelminthic drugNoYes 374342 286238 76.569.6 1.29 (1.01-1.65)1 0.038 N: Number examined; no: Number positive. Reference group marked as OR = 1; CI: Confidence interval. Significant association (p<0.05). * Variables were confirmed by multivariate analysis as significant predictors of IPIs. Discussion As shown by the results of the present study, intestinal parasitic infections (IPIs) are still a major public health problem (i.e., overall prevalence of 73.2%) among the impoverished and underprivileged communities in rural and remote West Malaysia. However, this study also observed some very encouraging trends. In Sungai Layau village where each family was provided with a concrete house and basic amenities like treated water supply, prevalence of IPIs was shown to be significantly lower (4.5%). This proved that proper provision of basic infrastructure and education are effective tools to reduce the prevalence of these infections. On the contrary, in Betau, Kuala Betis, Sungai Bumbun, Sungai Perah, Gurney, Pos Iskandar and Bukit Serok villages where some villagers still lived in traditional-built houses and using water from wells or rivers, prevalence of IPIs were very high. This was evident in the present findings whereby Betau village which was less provided or developed had the highest rate of infection (97.8%). Results also showed that STH infections (73.2%) were more common compared to protozoa infections (20.1%). T. trichiura infection is the most common (66.8%) followed by A. lumbricoides (38.5%) and hookworm (12.8%). These findings were in agreement with other previous local studies where T. trichiura infection was found to be the most prevalent (range: 26.0% to 98.2%), followed by A. lumbricoides infections (range: 19.0% to 67.8%) and lastly hookworm infections (range: 3.0% to 37.0%) [17], [18], [19], [20], [21]. However, global data has indicated that A. lumbricoides infections were the most prevalent among the three STH infections. The higher rate of T. trichiura infection has been reported to be due to the ineffective dosage and choice of anthelminthic used. Currently, the recommended treatment regime for STH infection is broad spectrum anthelminthics such as albendazole and mebendazole. Important therapeutic differences do exist between these drugs which affect their uses in clinical practice [22]. Both drugs are effective against ascariasis in single dose, whereas single doses of either albendazole or mebendazole have been found to be ineffective in many cases of trichuriasis [22]. Furthermore, potential resistance of T. trichiura to anthelminthic drugs has been highlighted in two intervention studies in Malaysian communities [23], [24]. It has been noted that unscheduled deworming without proper monitoring system was common among the children of these communities. Since the mass deworming program of schoolchildren has been discontinued in 1983 [25], some of the children received anthelminthic drug during visits to health clinic or from their school medical health team. Some parents have also bought anthelminthic drug for their children without following the recommended treatment intervals (i.e., periodic deworming) and this could have resulted in the inefficacy of the drug and subsequently lead to drug resistance [24]. Another important problem encountered in treatment is the high rate of re-infection especially in highly endemic areas. Local studies among rural communities have found that re-infection can occur as early as 2 months post treatment, by 4 months almost half of the treated population had been re-infected [24] and by 6 months the intensity of infections had returned to pre-treatment levels [26]. Similar findings have also been reported in other parts of the world indicating that by 6 months, the intensity of infections of T. trichiura and A. lumbricoides were similar to pre-treatment levels [27]. WHO has recommended that mass deworming programme should be carried out in communities when the cumulative STH prevalence is more than 50% or the cumulative percentage of moderately or heavily infected individuals is more than 10% [28]. As the present findings have indicated that the overall prevalence was 73.2%, it is strongly recommended that mass deworming programmes are restored and a systematic evaluation of treatment regime must be put in place to reduce the rates of re-infection. As for protozoa infections, the overall prevalence was 21.4%. However, in contrast with the latest local study in rural area, Noor Azian and colleagues reported very high rates of protozoa infection (72.3%) [29]. The present study found that G. duodenalis (10.4%) was the most predominant protozoa, followed by E. histolytica/dispar (10.2%) and lastly Cryptosporidium sp. (2.1%). In Malaysia, the prevalence of G. duodenalis infections varied from 2.0% to 29.2% while the prevalence of E. histolytica/dispar infections was reported to range from 1.0% to 18.5% among rural community [23], [29]. Although amoebic liver abscess (65% of 34) has been documented in patients admitted to an urban hospital in Malaysia [30], information from rural communities is not available as this infection can only be confirmed in a hospital setup. Two previous studies have indicated that Cryptosporidium sp. infections in rural areas ranged from 5.5% to 20.1% [31], [32]. The present study also reported 2 cases (0.3%) of Fasciolopsis/Fasciola sp. infection in Gurney village. This infection is most probably spurious due to consumption of infected animal liver. To date, there has not been any published data on intestinal fluke infection in West Malaysia, however, a case report of fasciolopsiasis by Fasciolopsis buski has been reported among rural community in East Malaysia [33]. In addition, two reported cases of food-borne diphyllobothriasis after consuming sushi and sashimi have also been reported in urban West Malaysia [34], [35]. Previous local studies indicated that there was a web of risk factors associated with the high prevalence of IPIs which included age, low family income, inadequate sanitation, presence and close contact with livestock or pets, untreated water supply, low level of parental education, poor geographical and personal hygiene [17], [22], [23]. Using multivariate analysis, the present study confirmed that children, low household income, untreated water supply, indiscriminate defecation were significant risk predictors of IPIs. This finding is further confirmed with the significantly lower prevalence in Sungai Layau village where household incomes are much higher and basic amenities provided by the government are fully utilized by the villagers. Conclusion Intestinal parasitic infections are highly prevalent and are major public health concerns among the poor and socioeconomically deprived rural and remote communities in West Malaysia. Given that IPIs are intimately associated with poverty, poor environmental sanitation and lack of clean water supply, it is crucial that these factors are addressed effectively. Improvement of socioeconomic status, sanitation, health education to promote awareness about health and hygiene together with periodic mass deworming are better strategies to control these infections. With effective control measures in place, these communities (especially children) will have a greater opportunity for a better future in terms of health and educational achievement. Supporting Information Checklist S1 STROBE checklist. (0.08 MB DOC) Click here for additional data file.
              Bookmark
              • Record: found
              • Abstract: found
              • Article: not found

              Reassessment of the epidemiology of amebiasis: state of the art.

              The epidemiology of amebiasis has dramatically changed since the separation of Entamoeba histolytica and Entamoeba dispar species, and the worldwide prevalence of these species has not been estimated until recently. The most cited data regarding prevalence, morbidity, or mortality due to amebiasis is the 1986 Walsh report, in which 100,000 deaths are reported to occur worldwide each year due to medical complications of invasive amebiasis. However, the prevalence values of Entamoeba histolytica infection could be completely erroneous since the estimations were performed prior to the molecular characterization of E. histolytica and E. dispar species. Moreover, Entamoeba moshkovskii, another morphologically indistinguishable human parasitic Entamoeba, was not mentioned or considered as a contributor to the prevalence figures in endemic areas. However, recent available prevalence and morbidity data obtained through molecular techniques allow the construction of a more reliable map of endemic regions of amebiasis all over the world [the Asian subcontinent (India, Bangladesh), Africa, Asian Pacific Countries (Thailand, Japan), South and Central America (Mexico, Colombia)]. The epidemiology of infectious diseases focuses on identification of factors that determine disease distribution in time and space, transmission factors responsible for the disease, clinical manifestations, and progression in the host, with the goal being the design of realistic intervention and prevention strategies in a reasonable period of time. In the present review, we will describe how molecular tools have made actual knowledge regarding the epidemiology of amebiasis possible. We will also analyze the most relevant available data on prevalence, morbidity, geographic distribution, patterns of transmission, exposure, and risk factors for infection in the human host. Our intention is to emphasize the recent molecular typing methods applied in genotyping Entamoeba species and strains, and to assess their value and limitations. Finally, we will discuss those areas of the host-parasite relationship that are still not fully understood, and the scientific challenges to approach this important public health problem in the future.
                Bookmark

                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS Negl Trop Dis
                PLoS Negl Trop Dis
                plos
                plosntds
                PLoS Neglected Tropical Diseases
                Public Library of Science (San Francisco, CA USA )
                1935-2727
                1935-2735
                3 December 2015
                December 2015
                : 9
                : 12
                : e0004225
                Affiliations
                [1 ]Department of Zoology, Gurucharan College, Silchar, Assam, India
                [2 ]Department of Biotechnology, Assam University, Silchar, Assam, India
                [3 ]School of Life Sciences, Jawaharlal Nehru University, New Delhi, India
                International Centre for Diarrhoeal Disease Research, BANGLADESH
                Author notes

                The authors have declared that no competing interests exist.

                Conceived and designed the experiments: BS JP. Performed the experiments: JN. Analyzed the data: BS JP SKG. Contributed reagents/materials/analysis tools: BS JP. Wrote the paper: JN JP BS.

                Article
                PNTD-D-15-00970
                10.1371/journal.pntd.0004225
                4669114
                26633890
                eceb0610-fd21-4255-8589-40227e90e562
                © 2015 Nath et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited

                History
                : 5 June 2015
                : 21 October 2015
                Page count
                Figures: 4, Tables: 5, Pages: 19
                Funding
                BS and JP received the joint funding from the Department of Biotechnology, Ministry of Science and technology, New Delhi, India through North East Twining program. Grant number BT/55/NE/TBP/2010 to carry out this project. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Custom metadata
                All relevant data are within the paper and its Supporting Information files.

                Infectious disease & Microbiology
                Infectious disease & Microbiology

                Comments

                Comment on this article