Control Of Schistosomiasis Japonica Research Articles

A chromosome-level genome of the human blood fluke Schistosoma japonicum identifies the genomic basis of host-switching.

The evolution and adaptation of S.japonicum, a zoonotic parasite that causes human schistosomiasis, remain unclear because of the lack of whole-genome data. We construct a chromosome-level S.japonicum genome and analyze it together with 72 samples representing six populations of the entire endemic region. We observe a Taiwan zoophilic lineage splitting from zoonotic populations ∼45,000 years ago, consistent with the divergent history of their intermediate hosts. Interestingly, we detect a severe population bottleneck in S.japonicum, largely coinciding with human history in Asia during the last glacial maximum. We identify several genomic regions underlying natural selection, including GATAD2A and Lmln, both showing remarkable differentiation among different areas. RNAi knockdown suggests association of GATAD2A with parasite development and infection in definitive hosts, while Lmln relates to the specificity of the intermediate hosts. Our study provides insights into the evolution of S.japonicum and serves as a resource for further studies.

The status of schistosomiasis japonica control in the Philippines: The need for an integrated approach to address a multidimensional problem

Schistosomiasis japonica remains a public health concern in many areas of the Philippines. Adequate and updated information is essential to enhance policy and service delivery toward control and elimination. Despite the efforts on schistosomiasis control in the Philippines, some challenges remain in these dimensions. An integrated surveillance system is recommended to determine the prevalence of infection in humans, animal reservoirs, and snail intermediate hosts, allowing the identification of high-priority areas for targeted interventions. This will entail the enhancement of laboratory diagnosis capacity through the use of more sensitive techniques, complemented by capacity building of concerned human and animal health professionals. Given the zoonotic nature of schistosomiasis japonica, adopting the One Health approach is essential to influence policies and interventions that may accelerate control and elimination. This can be achieved through the attainment of mass drug administration coverage targets and intensified case finding and management, robust implementation and integration of veterinary public health activities, the conduct of snail control measures, provision of safe water, sanitation, and hygiene services, and health promotion and education into the national schistosomiasis control and elimination program. This review aimed to describe the status of schistosomiasis japonica control in the Philippines in the context of human health, animal health, vector ecology and management, environmental health, and sociocultural dimensions.

Qualitative and quantitative proteomic analyses of Schistosoma japonicum eggs and egg-derived secretory-excretory proteins

BackgroundSchistosome parasites lay up to a thousand eggs per day inside the veins of their mammalian hosts. The immature eggs deposited by females against endothelia of venules will embryonate within days. Approximately 30% of the eggs will migrate to the lumen of the intestine to continue the parasite life-cycle. Many eggs, however, are trapped in the liver and intestine causing the main pathology associated with schistosomiasis mansoni and japonica, the liver granulomatous response. Excretory-secretory egg proteins drive much of egg-induced pathogenesis of schistosomiasis mansoni, and Schistosoma japonicum induce a markedly distinct granulomatous response to that of S. mansoni.MethodsTo explore the basis of variations in this responsiveness, we investigated the proteome of eggs of S. japonicum. Using mass spectrometry qualitative and quantitative (SWATH) analyses, we describe the protein composition of S. japonicum eggs secretory proteins (ESP), and the differential expression of proteins by fully mature and immature eggs, isolated from faeces and ex vivo adults.ResultsOf 957 egg-related proteins identified, 95 were exclusively found in S. japonicum ESP which imply that they are accessible to host immune system effector elements. An in-silico analysis implies that ESP are able of stimulating the innate and adaptive immune system through several different pathways. While quantitative SWATH analysis revealed 124 proteins that are differentially expressed by mature and immature S. japonicum eggs, illuminating some important aspects of eggs biology and infection, we also show that mature eggs are more likely than immature eggs to stimulate host immune responses.ConclusionsHere we present a list of potential targets that can be used to develop better strategies to avoid severe morbidity during S. japonicum infection, as well as improving diagnosis, treatment and control of schistosomiasis japonica.

Publication output of the new integrated strategy for schistosomiasis japonica control in China: a PubMed-based bibliometric assessment

Abstract Background Although sustainable control since 1950s has achieved great successes, schistosomiasis japonica remains a major public health problem in China. Since 2004, a new integrated strategy was developed aiming to control the transmission of Schistosoma japonicum through the implementation of a package of interventions. To date, no systematic review or meta-analysis assessing the effectiveness of this new integrated strategy for schistosomiasis control in China has been published. We performed a PubMed-based bibliometric assessment of publications on the new integrated strategy for schistosomiasis japonica control in China, to understand the global transmissibility and sharing of the new integrated strategy. Methods An in-depth bibliometric analysis of all publications on the new integrated strategy for schistosomiasis japonica control in China was performed through a PubMed search using the terms “schistosomiasis” and “China,” from January 1, 2004 to August 31, 2018. All titles and abstracts were read carefully, and the publications reporting the effectiveness, experiences, lessons, or problems of the new integrated strategy were included in the bibliometric analysis. Results Overall, 2,361 titles were screened, and 70 eligible publications were accessed for analyses, including 23 studies in English, published in 15 international journals, and 47 studies in Chinese with abstracts in English, published in 3 national journals. Chinese Journal of Schistosomiasis Control (Zhongguo Xue Xi Chong Bing Fang Zhi Za Zhi) published 60% of the research output. Research articles (48.6%) and short reports (37.1%) were the dominant manuscript types. Furthermore, 471 contributing authors from 277 affiliations across 9 countries produced these 70 publications. Conclusion This is the first PubMed-based quantitative analysis of the research output of the new integrated strategy, and our data indicate a low global transmissibility of Chinese new integrated strategy. We therefore call for more research outputs of the new integrated strategy for schistosomiasis japonica control in China to be communicated through international platforms.

Schistosomiasis Japonica Control in Domestic Animals: Progress and Experiences in China.

Schistosomiasis japonica, caused by Schistosoma japonicum, is an endemic, zoonotic parasitic disease. Domestic animals, particularly bovines, are thought to play an important role in transmission of the disease. Historically, China was the country mostly severely impacted by schistosomiasis japonica, but now prevalence and morbidity have been greatly reduced. Since the mid-1950s when China launched the National Schistosomiasis Control Program, the control of schistosomiasis in domestic animals has been carried out almost synchronously with that of human schistosomiasis, and this concept has been proven to be effective. Generally, the campaign of schistosomiasis japonica control in domestic animals in China went through four phases over the past six decades, namely, the large-scale epidemiological investigation phase, the case screening and small-scale chemotherapy phase, the mass chemotherapy phase, and the infection source control phase. These distinct phases were responsive to changing disease epidemiology, socioeconomic development, and technological advances, resulting in successful attainment of disease control goals.

Long-term effectiveness of the integrated schistosomiasis control strategy with emphasis on infectious source control in China: a 10-year evaluation from 2005 to 2014.

Schistosomiasis is a neglected tropical parasitic disease of great public health significance worldwide. Currently, mass drug administration with praziquantel remains the major strategy for global schistosomiasis control programs. Since 2005, an integrated strategy with emphasis on infectious source control was implemented for the control of schistosomiasis japonica, a major public health concern in China, and pilot studies have demonstrated that such a strategy is effective to reduce the prevalence of Schistosoma japonicum infection in both humans and bovines. However, there is little knowledge on the long-term effectiveness of this integrated strategy for controlling schistosomiasis japonica. The aim of this study was to evaluate the long-term effectiveness of the integrated strategy for schistosomiasis control following the 10-year implementation, based on the data from the national schistosomiasis control program released by the Ministry of Health, People's Republic of China. In 2014, there were 5 counties in which the transmission of schistosomiasis japonica had not been interrupted, which reduced by 95.2% as compared to that in 2005 (105 counties). The number of schistosomiasis patients and acute cases reduced by 85.5 and 99.7% in 2014 (115,614 cases and 2 cases) as compared to that in 2005 (798,762 cases and 564 cases), and the number of bovines and S. japonicum-infected bovines reduced by 47.9 and 98% in 2014 (919,579 bovines and 666 infected bovines) as compared to that in 2005 (1,764,472 bovines and 33,736 infected bovines), respectively. During the 10-year implementation of the integrated strategy, however, there was a minor fluctuation in the area of Oncomelania hupensis snail habitats, and there was only a 5.6% reduction in the area of snail habitats in 2014 relative to in 2005. The results of the current study demonstrate that the 10-year implementation of the integrated strategy with emphasis on infectious source has greatly reduced schistosomiasis-related morbidity in humans and bovines. It is concluded that the new integrated strategy has remarkable long-term effectiveness on the transmission of schistosomiasis japonica in China, which facilitates the shift of the national schistosomiasis control program from transmission control to transmission interruption and elimination. However, such a strategy seems to have little effect on the shrinking of areas of snail habitats.

Evaluating the effect of a novel molluscicide in the endemic schistosomiasis japonica area of China.

Oncomelania hupensis is the sole intermediate host snail of Schistosoma japonicum in China. Snail control by molluscicide remains one of the most effective measures of schistosomiasis japonica control. A 50% wettable powder of niclosamide ethanolamine salt (WPN) is widely used for snail control in China. However, WPN is costly and toxic to fish. A novel molluscicide named LDS, the salt of quinoid-2′, 5-dichloro-4′-nitrosalicylanilide from niclosamide, has been developed. To evaluate the effects of large-scale field application of LDS on field snail control, tests were conducted in 15 counties of Hubei Province, China. Active adult snails, were immersed in 0.2, 0.4, and 0.6 g/m3 of 10% LDS, 1.0 g/m3 of 50% WPN was used as the molluscicide control, and then the mortality rates of snails were investigated after 1, 2, and 3 days. In addition, four active concentrations of 10% LDS (0.4, 0.6, 0.8 and 1.0 g/m2) were applied by spraying and powdering in the field. 1.0 g/m2 of 50% WPN was used as the molluscicide control, and then the mortality rates of snails were observed after 1, 3, and 7 days. The results indicated that 0.4 g/m3 LDS applied by the immersion or 0.6 g/m2 LDS applied by spraying and powdering achieved the same molluscicidal effect as that of WPN, regardless of exposure time. By using different methods, the snail mortality rates in the molluscicide groups were related to exposure time and concentration, respectively. LDS costs less than WPN; thus, LDS is suitable and applicable for use as a molluscicide in schistosomiasis japonica epidemic areas.

Spatio-temporal analysis to identify determinants of Oncomelania hupensis infection with Schistosoma japonicum in Jiangsu province, China

BackgroundWith the successful implementation of integrated measures for schistosomiasis japonica control, Jiangsu province has reached low-endemicity status. However, infected Oncomelania hupensis snails could still be found in certain locations along the Yangtze river until 2009, and there is concern that they might spread again, resulting in the possible re-emergence of infections among people and domestic animals alike. In order to establish a robust surveillance system that is able to detect the spread of infected snails at an early stage, sensitive and reliable methods to identify risk factors for the establishment of infected snails need to be developed.MethodsA total of 107 villages reporting the persistent presence of infected snails were selected. Relevant data on the distribution of infected snails, and human and livestock infection status information for the years 2003 to 2008 were collected. Spatio-temporal pattern analysis including spatial autocorrelation, directional distribution and spatial error models were carried out to explore spatial correlations between infected snails and selected explanatory factors.ResultsThe area where infected snails were found, as well as their density, decreased significantly between 2003 and 2008. Changes in human and livestock prevalences were less pronounced. Three statistically significant spatial autocorrelations for infected snails were identified. (i) The Moran’s I of infected snails increased from 2004 to 2007, with the snail density increasing and the area with infected snails decreasing. (ii) The standard deviations of ellipses around infected snails were decreasing and the central points of the ellipses moved from West to East. (iii) The spatial error models indicated no significant correlation between the density of infected snails and selected risk factors.ConclusionsWe conclude that the contribution of local infection sources including humans and livestock to the distribution of infected snails might be relatively small and that snail control may limit infected snails to increasingly small areas ecologically most suitable for transmission. We provide a method to identify these areas and risk factors for persistent infected snail presence through spatio-temporal analysis, and a suggested framework, which could assist in designing evidence based control strategies for schistosomiasis japonica elimination.

Application of praziquantel in schistosomiasis japonica control strategies in China

Schistosomiasis remains a major public health problem with an estimated 200 million people infected in the world, and in China, schistosomiasis japonica is endemic in the south part of the country. In 1960s, before praziquantel was developed, there were about seven million patients. Praziquantel has a high efficacy against Schistosoma, few and transient side effects, simple administration and competitive cost, and is equally suited for both individual and large-scale treatment. Praziquantel has been widely used in the morbidity control, transmission control, and prevention of schistosomiasis japonica in China since 1980s. The schemes of praziquantel chemotherapy include the diagnostic selective chemotherapy, extensive chemotherapy, mass chemotherapy, stratified chemotherapy, phased chemotherapy, etc. Chemotherapy alone or combined with other control measure, such as Oncomelania snail control, health education, safety water supply, and so on, has achieved a great success, and there are only 0.33 million infected people now. This paper reviews the application of praziquantel in the schistosomiasis japonica control strategies in China so as to provide the rich experiences for reference of health workers of other countries where schistosomiasis is endemic.

Differences in genomic architecture between two distinct geographical strains of the blood fluke Schistosoma japonicum reveal potential phenotype basis

The Chinese (SjC) and Philippine (SjP) strains of the blood fluke Schistosoma japonicum have been shown to present clearly different phenotypes in fecundity, pathology, drug sensitivity and immunology. We used microarray based comparative genomic hybridisation (aCGH) to investigate structural differences in the genomes of the two strains and identified seven distinct regions of the S. japonicum genome that present differential aCGH representing either deletion or duplication regions in SjP. Within these regions, genes predicted to be associated with the recognised phenotypic differences were identified and that may provide new insights into the biology and evolution of the two strains, with implications for the epidemiology and control of schistosomiasis japonica in China and the Philippines.

Effectiveness of a comprehensive schistosomiasis japonica control program in Jiangsu province, China, from 2005 to 2008

The effectiveness of a comprehensive schistosomiasis japonica control program implemented in 8 villages along the Yangtze river in Jiangsu province from 2005 to 2008 was studied. Control measures included snail control, chemotherapy of humans and livestock, health education, and transmission cycle interruption using sanitation in dwellings and at anchorage sites for fishermen and sailors. The Schistosoma japonicum prevalence among residents and livestock, the total area of snail habitats, the Oncomelania hupensis snail density, and the percentage of infected snails served as indicators for the effectiveness of the control efforts. After 4 years of program implementation, the seroprevalence in humans had decreased from 9.03% to 3.24% (P<0.001) and the parasitological prevalence among males had decreased from 0.42% to 0.12% (P=0.004). Among females, it remained stable at a low level. The S. japonicum prevalence in livestock had decreased from 2.94% to 0% (P<0.001). Additionally, the area where infected snails could be found had shrunk from 89.99 hectares (ha) to 16.00ha, the snail density had decreased from 0.56 to 0.32 per 0.1m2, and the percentage of infected snails had dropped from 0.38% to 0.12% (all P<0.001). The results demonstrate that an integrated schistosomiasis japonica control strategy focusing on the main transmission cycles and reservoirs and combines chemotherapy, infrastructure interventions and health education combined with robust surveillance is feasible and allows to effectively control S. japonicum.

Conquering ‘snail fever’: schistosomiasis and its control in China

Schistosomiasis japonica is a serious parasitic disease and a major health risk for more than 60 million people living in the tropical and subtropical zones of south China. The disease is a zoonosis and its cause, the parasitic trematode Schistosoma japonicum, has a range of mammalian reservoirs, making control efforts difficult. Current control programs are heavily based on community chemotherapy with a single dose of the highly effective drug praziquantel. However, vaccines (for use in bovines and in humans) in combination with other control strategies are needed to eliminate the disease. In this review, we provide an overview of the transmission, clinical features, pathogenesis, diagnosis, treatment, genetics and susceptibility, epidemiology, and prospects for control of schistosomiasis japonica in China. The threat posed by the Three Gorges Dam may undermine control efforts because it will change the local ecology and associated schistosomiasis transmission risks over the next decade and beyond.

Effects of the praziquantel-based control of schistosomiasis japonica in China

In 2000-2002, the effects of the praziquantel-based control of Schistosoma japonicum (in which each subject found infected in the October of each year is given a single dose of the drug) were examined at 20 or 21 surveillance sites across China. The pre- and post-treatment prevalences and intensities of infection were compared at each site and after pooling using a random-effects method. One year after treatment, in the communities that had the higher prevalences of infection (> 10%) before treatment, the mean prevalence of infection was found to have fallen by 20.15% [95% confidence interval (CI)=6.95%-31.48%] and the geometric mean intensities of infection (measured as eggs/g faeces) in the currently infected individuals and the entire study cohort were found to have fallen, by 22.91% (CI=14.69%-30.34%) and 33.93% (CI=11.69%-50.68%), respectively (P<0.05 for each). In the communities that had lower prevalences of infection pre-treatment, however, no statistically significant reductions in the intensity of infection were observed, although the prevalences in the communities that had pre-treatment prevalences of 6%-10% did fall significantly post-treatment, by a mean of 24.50% (CI=5.35%-39.83%; P<0.05). These observations indicate that the current annual surveys for infection, in which those found infected are each given a single dose of praziquantel, should help keep the prevalence of human infection with S. japonicum low in China, although chemotherapy alone is unlikely to eradicate the parasite.

Strategy formulation for schistosomiasis japonica control in different environmental settings supported by spatial analysis: a case study from China

With the aim of exploring the usefulness of spatial analysis in the formulation of a strategy for schistosomiasis japonica control in different environmental settings, a population-based database was established in Dangtu county, China. This database, containing the human prevalence of schistosomiasis at the village level from 2001 to 2004, was analyzed by directional trend analysis supported with ArcGIS 9.0 to select the optimum predictive approach. Based on the approach selected, different strata of prevalence were classified and the spatial distribution of human infection with Schistosoma japonicum was estimated. The second-order ordinary kriging approach of spatial analysis was found to be optimal for prediction of human prevalence of S. japonicum infection. The mean prediction error was close to 0 and the root-mean-square standardised error was close to 1. Starting with the different environmental settings for each stratum of transmission, four areas were classified according to human prevalence, and different strategies to control transmission of schistosomiasis were put forward. We conclude that the approach to use spatial analysis as a tool to predict the spatial distribution of human prevalence of S. japonicum infection improves the formulation of strategies for schistosomiasis control in different environmental settings at the county level.

Development of a vaccine against Schistosoma japonicum in China: a review

Significant progress has been made over the past 50 years in the control of schistosomiasis japonica in China. However, recent data suggest that the disease is re-emerging. By the end of 2003, Schistosoma japonicum was still endemic in 110 counties in seven provinces in the southern part of China where the long-term reduction of the disease has been replaced by an increase in the number of people infected and areas infested by the intermediate host snail, i.e. Oncomelania hupensis. Explanations are multifactorial, including the construction of the Three Gorges dam, major flooding events, recovery of the Dongting Lake and the possible impact of climate change. An efficacious vaccine against S. japonicum would represent a significant addition to the current arsenal of control tools, particularly in the framework of an integrated control approach. The vaccine could be targeted either towards the prevention of infection or towards the reduction of parasite fecundity. Although progress in this field has been relatively slow, encouraging results have been obtained in recent years using defined native and recombinantly derived S. japonicum antigens. These findings suggest that development of a safe and efficacious vaccine is feasible. This paper reviews the progress in the development of a vaccine against S. japonicum in China, and includes also data from foreign researchers who are engaged in collaborative work with Chinese scientists.

Transmission control of schistosomiasis japonica: implementation and evaluation of different snail control interventions

The great progress made in the control of schistosomiasis japonica in China is to some extent explained by successful intermediate host snail control, in particular with environmental management commencing some 50 years ago. By 1995, interruption of Schistosoma japonicum transmission had been achieved in five of the 12 schistosome-endemic provinces while endemic areas in the remaining provinces had been reduced significantly, and snail habitats had decreased by 74%. In this paper, we review the role and approaches towards control and give an account of compounds toxic to snails. Mid- and long-term national plans for schistosomiasis control are discussed, including integrated snail control approaches as an important measure. It is anticipated that implementation of these plans will accelerate snail control, which in turn will consolidate and mutually reinforce other control measures, most notably praziquantel-based chemotherapy. The ultimate aim is to eliminate schistosomiasis as a public health problem in China.

Use of praziquantel for clinical treatment and morbidity control of schistosomiasis japonica in China: a review of 30 years’ experience

Chemotherapy is widely acknowledged as the most important, rapid and cost-effective method of reducing morbidity due to schistosome infections. The discovery of praziquantel in the 1970s has been a breakthrough for treatment of patients infected with schistosomes, including Schistosoma japonicum in China, and entire communities at risk of schistosomiasis. Praziquantel is usually administered in a single oral dose and has no or only mild and transient side effects. The drug is highly efficacious against S. japonicum, both in patients with acute and chronic stages of the infection, among subjects with extensive hepatosplenic involvement, and in patients with other complicated diseases. The cost of praziquantel has been reduced significantly over the past years. Hence, praziquantel has become the backbone of the national schistosomiasis control programme in China and in other countries where the disease remains endemic, most notably in sub-Saharan Africa. Chemotherapy with praziquantel also plays a role in transmission control of schistosomiasis, although transmission interruption cannot be reached by chemotherapy alone. Here, I review 30 years’ of experiences gained with the use of praziquantel for clinical treatment and larger-scale control of schistosomiasis japonica in China.

A double-blind field trial on the effects of artemether on Schistosoma japonicum infection in a highly endemic focus in southern China

To further strengthen the evidence-base of artemether for the control of schistosomiasis japonica, a randomised controlled trial was carried out in the Poyang Lake region, a highly endemic area in southern China. A total of 783 individuals, aged 6–60 years, were enrolled. They were first given a single oral dose of praziquantel (50 mg/kg). Then, they were randomly assigned oral artemether (6 mg/kg) or placebo, administered once every 2 weeks for 9–11 doses, covering the entire transmission season for Schistosoma japonicum in 2004. Stool examination 1 month after the final dosing revealed eggs of S. japonicum in 3/373 (0.8%) of the artemether recipients and 56/361 (15.0%) in placebo recipients ( χ 2 = 53.69, P < 0.001). Compared to the baseline, the geometric mean intensity of S. japonicum infection had decreased by 96.1% in the artemether group, and increased by 50.8% in the placebo group. No acute cases of schistosomiasis japonica were observed in the artemether group, whereas three such cases were reported from the placebo group. Compliance with regard to multi-doses of artemether and placebo was 84.9, and 77.9%, respectively. This study confirms that repeated oral artemether produces no drug-related adverse effects, significantly reduces incidence and intensity of patent S. japonicum infection and results in high compliance. Hence it can be used as an additional tool for the control of schistosomiasis japonica in the lake regions of China.

A Bayesian-based approach for spatio-temporal modeling of county level prevalence of Schistosoma japonicum infection in Jiangsu province, China

Spatio-temporal variations of Schistosoma japonicum infection risk in Jiangsu province, China, were examined and the relationships between key climatic factors and infection prevalence at the county level were determined. The parasitological data were collected annually by means of cross-sectional surveys carried out in 47 counties from 1990 to 1998. Climatic factors, namely land surface temperature (LST) and normalized difference vegetation index (NDVI), were obtained from remote sensing satellite sensors. Bayesian spatio-temporal models were employed to analyze the data. The best fitting model showed that spatial autocorrelation in Jiangsu province decreased dramatically from 1990 to 1992 and increased gradually thereafter. A likely explanation of this finding arises from the large-scale administration of praziquantel for morbidity control of schistosomiasis. Our analysis suggested a negative association between NDVI and risk of S. japonicum infection. On the other hand, an increase in LST contributed to a significant increase in S. japonicum infection prevalence. We conclude that combining geographic information system, remote sensing and Bayesian-based statistical approaches facilitate integrated risk modeling of S. japonicum, which in turn is of relevance for allocation of scarce resources for control of schistosomiasis japonica in Jiangsu province and elsewhere in China, where the disease remains of public health and economic significance.

Gene cloning and complete nucleotide sequence of philippine Schistosoma japonicum paramyosin

The development of an effective vaccine is recognised as a necessary adjunct to the control of schistosomiasis japonica, a disease affecting several million people in China and the Philippines. Currently, recombinant Schistosoma japonicum molecules are considered most suitable for large scale vaccine production and a number of genes encoding vaccine candidate polypeptides have been cloned and expressed (see Waine et al., 1993a). One of the molecules providing most promise as a vaccine target is paramyosin (Butterworth, 1992), a major structural protein of thick filaments in the muscle of most invertebrates; paramyosin genes have now been cloned from a range of parasitic helminths, including schistosomes (Limberger and McReynolds, 1990; Laclette et al., 1991; Dahmen et al., 1993; Landa et al., 1993; Mühlschlegel et al., 1993, Nara et al., 1994). The cloning and nucleotide sequence of S. Japonicum paramyosin is described.