Anopheles minimus complex (Cellia: Myzomyia), An. minimus and Anopheles harrisoni, respectively former species A and C. Because the morphological identification of both sibling species is difficult and may lead to misidentifications, accurate data on their respective distribution are missing. This is of fundamental importance since the two species seem to exhibit differential vectorial capacities for malaria transmission.The distribution of anopheline mosquitoes in Vietnam was examined, with a particular interest for the two sibling species of the An. minimus s.l. were carried out in 23 sites throughout northern, central and south-eastern regions of Vietnam.Large entomological surveys based on cattle collections and molecular identifications of An. minimus and An. harrisoni were mapped. It is noteworthy that there was a high specific biodiversity at each study site. Anopheles minimus s.l. and Anopheles sinensis were the main anopheline species in the northern region, whereas Anopheles aconitus and Anopheles vagus were the most frequent ones in the central region. The southern limit of An. harrisoni was increased to the latitude of 11°N. Sympatry between both sibling species has been extended to new provinces.Based on previous molecular works and our data, the distribution of anopheline species and the relative densities of Malaria transmission is still high in central Vietnam and along bordering countries. Therefore, it is important to know and map the precise distribution of the main and secondary malaria vectors in Vietnam for applying efficient vector control programmes. Moreover, these maps should be regularly updated and linked to environmental characteristics relative to disease epidemiology, and environmental and climatic changes occurring in southeast Asia. Anopheles minimus s.l.  is composed of three sibling species, An. minimus (former species A), Anopheles harrisoni (former species C), and An. minimus species E [Anopheles minimus and An. harrisoni can be sympatrically distributed over the southeast Asian mainland whereas species E is restricted to the Ishigaki Island in the Ryukyu Archipelago, Japan, a malaria free region [An. minimus and An. harrisoni are missing [Anopheles aconitus, Anopheles pampanai and Anopheles varuna, are common [The main malaria vector pecies E . Anophele region ,3. Becaue region , accurate common ,6. Recene common -10.Anopheles minimus s.l. occurs in hilly, forested areas [An. minimus s.l. are streams or canals, with slow-running water partially shaded by grassy margins [An. minimus s.l. and Anopheles dirus s.l. occur [ed areas . Norther margins . In centl. occur ,11. The l. occur ,13.An. minimus was molecularly reported in 17 provinces; and An. harrisoni confirmed in nine provinces  to compile a distribution map for the two members of the Anopheles minimus complex, based on previous records and 23 new study sites.Since these sibling species may exhibit different vectorial capacities ,15, it iA total of 23 sites in 15 provinces were selected for mosquito collections in different geographical areas of northern, central and south-eastern Vietnam  Table . MosquitAn. minimus, An. harrisoni and associated anopheline species . One aconitus , may havaconitus . The An.Based on morphological identifications, a total of 19 anopheline species were collected and identified throughout the northern to south-eastern regions of Vietnam Table  out of tAn. minimus s.l. were DNA-extracted and identified as An. minimus or An. harrisoni for all the sites, except five sites with samples not available and three sites with no An. minimus s.l. populations (Table An. minimus samples (two An. minimus s.l instead of two An. aconitus) were noticed for the VSLA site (northern region). No sites with An. harrisoni only were found neither for the northern region nor the central one . This may reflect a competition between these two sibling species which needs to be further investigated. Molecular species identifications were intentionally not carried out for the Dirus and Maculatus complexes, because the Minimus complex was targeted. Recently, Obsomer et al.[A total of 706 ns Table . Only twe Figure . Intereser et al. publisheAnopheles minimus s.l. and An. sinensis were the main species collected on cattle over the northern region, followed by An. aconitus and An. jeyporiensis  and one site (VTHB) presented a very low density . Actually in Vietnam, An. minimus s.l. is commonly found at an altitude ranging from 200 to 800 meters and becomes quite rare at altitudes above 1,500 meters [An. minimus s.l. in the VTHB site (only three specimens) may be due to very recent agricultural changes linked to modified irrigation systems that may have disturbed the mosquito larval habitats, phenomenon also observed in Thailand [An. sinensis and Anopheles philippinensis, the latter species having much lower densities in all the other sites and is known to occur in agricultural areas [The VNAA village is surrounded by pine plantations. The overall number of anophelines collected was low (204 specimens during six nights) and itude 1,26 m. Actu0 meters . The lowThailand  and IndiThailand . The maial areas .An. harrisoni , whereas An. minimus was present in 13 out of 15 surveyed sites. Anopheles harrisoni was the predominant species of the complex in three sites only, two in the Son La Province (VSLA and VSLB), and one in the Lang Son Province (VLSB).Five sites out of 15 were negative for et al.[An. minimus only in their two sites in Lao Cai Province. However, one site (VLCA) was found positive for both sibling species in this province. Further south, the two sites VTHA and VNAB showed two different situations in the hilly areas of Thanh Hoa and Nghe An Provinces, respectively sympatric populations and An. minimus only.Regarding the distribution reported in previous studies in northern Vietnam Figure , the reset al. reportedAn. minimus and possibly An. harrisoni, represents a permanent threat for the resurgence of malaria in this region with the movement of infected human populations from endemic areas such as central Vietnam and bordering countries.In northern Vietnam, malaria is relatively well controlled but still occurs at the border of China and Laos . HoweverAnopheles aconitus and An. vagus were the most frequent species in these regions (Table Anopheles dirus s.l. was found in low densities in two sites only (VGLA and VPYA) (Table ns Table . AnophelA) Table . It occuA) Table .An. minimus s.l. and four sites among the seven had low densities ranging between one and six specimens (Table Anopheles harrisoni was recently found in Khanh Hoa Province [An. minimus which was reportedly the dominant species until 1999 [An. harrisoni to Binh Duong Province (VBGA), located in south-eastern Vietnam, at a latitude of 11°N, less than 100 km northwest of Ho Chi Minh City. This is a new site where An. minimus and An. harrisoni are sympatric.One site (VKTA) was negative for ns Table . AnophelProvince   and Thailand [am Table . Anophell region -27, and Thailand ,25,28,29An. minimus s.l. and An. sinensis were the main species in northern Vietnam, whereas An. aconitus and An. vagus were dominant in central Vietnam. Anopheles minimus and An. harrisoni of the Minimus complex are present over the northern, central and south-eastern Vietnam, down to latitude 11°N. Malaria transmission is still high in central Vietnam and along bordering countries. Future entomological surveys in the surrounding countries and, on a larger scale throughout southeast Asia, are required to molecularly identify the different members of the Minimus and Aconitus Subgroups to clarify the precise distributions of each member and to improve vector control strategies.Based on cattle collections, Compiling distribution maps based on large sample collections is not an academic exercise. Knowing precise and accurate geographic distributions of vector species is an important prerequisite for: (1) an adequate choice of zones where vector control actions should focus, (2) a better selection of future study sites for entomologists working on secondary or local vector species, (3) studies on anopheline biodiversity relating to environmental and climatic changes, (4) analyses of landscape-species associations, and (5) modelling malaria risk maps or comparison of predictive ecological maps with field observations.Since malaria transmission still occurs in central Vietnam and population movement from the centre to the north are important, vector control measures must be maintained. In addition, entomological, epidemiological and climatic data need to be integrated into a Geographic Information System to follow the future trends of the disease and to assess the malaria risk zones. These maps should be regularly updated and linked to environmental characteristics of disease epidemiology relating to environmental and climatic changes occurring in southeast Asia.The author(s) declare that they have no competing interests.CG participated in the spatial and data collections in northern Vietnam, carried out the molecular identification of the Minimus samples, analysed and interpreted the data, and drafted the manuscript. CVN coordinated the spatial data collections in northern and central Vietnam and made substantial intellectual contributions to the paper. THD coordinated all the entomological surveys and contributed to the content of the paper. MC  and SM initiated the study and revised the paper critically for important intellectual content. All authors read and approved the final manuscript.


To view the changing perspective of Psychiatry in Rajasthan, we must begin with the story of establishment of lunatic asylums in Rajputana states. It was after the treaty of alliance signed between the native states of Rajputana and the East India Company (1803-1818) that each native state created/established an asylum for detention and care of the lunatics. Initially the lunatics were seen in dispensaries and detained in jails tied with chains. An asylum outside the jail was first established in 1881 in Jaipur state. Ajmer had an asylum inside jail in 1865 (Hendley 1900). Forty unfortunate lunatics were incarcerated inside ‘cells’ in this asylum, which was situated near Ghatgate (Jaipur). In year 1894, another enclosure in a walled garden near Central Telegraph Office, Jaipur in the southern part of the city was used as asylum by Jaipur state. Here the lunatics were left to enjoy the Open Court Yard and a ‘Baradaree’, no cots were provided but the atmosphere was more humane and sympathetic towards the unfortunate lunatics who were confined in this asylum. Food and protection were provided by the jail department till 1922, when this asylum was shifted to a Dharamshala situated near Chandpole Gate opposite the present Janana Hospital. Here the lunatics were again put in cells. The medical supervision was provided by the part-time doctors from the jail department. Administration, watch and ward of asylum remained under the jail department till year 1943 .In 1943, Jaipur state enacted Jaipur Lunacy Act for detention and treatment of lunatics; the criminal procedure code 1926. Jaipur State was amended for the detention and care of the criminal lunatics 1943 (C.P.U. 1943). From the year 1944, the Mental Hospital Jaipur (Lunatic Asylum) started running as an independent hospital under Medical Department of Jaipur State. Till 1922, the control of lunatic asylum was placed under the Rajputana Agency, but after 1922 the administration of lunatic asylum was transferred to Jaipur Durbar. The annual returns from these asylums show a high mortality of lunatics admitted there in, on account of diarrhea, dysentery and fever. Under orders from Jaipur Durbar, the lunatics could be released from asylums for the care of relatives and friends if assured for kind sympathetic care and protection from doing harm to others. All lunatics were admitted under court orders.th century that psychoanalytic explanations came in and the last four decades have seen tremendous scientific advancements in understanding of human behavior through biopsychosocial research. The treatment of mental disorders has also been revolutionized in the last four decades and as a result to this the specialty is now a very fast growing subject.The discipline of psychiatry is rather a new specialty of medical science. Though understanding of human mind has existed from times immemorial in different philosophical and religious schools, in the middle ages, due to lack of knowledge, superstitions like evil spirits dominated the explanation for various mental disorders. It is only in the late 19As a result of research in mental health, now intricate complexities of the human mind have been understood in terms of neuroelectrical and neurochemical changes occurring in the brain. Specific change occurring with different mood states has been identified and an intervention to bring desired changes in human behavior is now possible. Mental illnesses once thought incurable are now treatable. Mental illness is now considered like any other illness and there is no reason that it should be concealed, and like patients with physical disorder the mentally ill after treatment can be rehabilitated and made useful citizens.Psychiatric Centre, Jaipur is the premier institute of the state for treatment and care of mentally ill. Founded in 1952, the institution has completed 53 years of its existence. Like the discipline of Psychiatry, the initial development was rather slow. But after its recognition as a postgraduate teaching department of the medical college in the year 1980, development has been seen in certain areas of its growth.It is one of the six attached hospitals of S.M.S. Medical College, Jaipur. The center was started with a bed strength of 180 with an outpatient attendance of 190 per annum and annual turnover of 105 indoor patients in the year 1952. At present the hospital has a total bed strength of 320 . The hospital has an annual turnover of average 4000 indoor patients and an outpatients' attendance of nearly 40,000 per annum.The catchment area of the hospital is whole of Rajasthan and adjacent states of U.P., Madhya Pradesh, Haryana, Punjab and State of Delhi. The admission facilities in the center are free to every person irrespective of the state. The medicines are distributed free of cost to all the indoor as well as outdoor patients.A 20-bedded emergency ward was started in the year 1981 for the patients requiring urgent care and management. The admission facility in the emergency ward is available round the clock. A separate 12-bedded de-addiction ward was also started in the year 1986 to meet the special needs of the drug-addicts.The outpatients department of the Center is run by consultants, civil assistant surgeons and postgraduate students. At present there are three Associate Professors, six Assistant Professors, five CAS and nine postgraduate students. There are posts for two clinical psychologists and two psychiatric social workers, who collaborate with the professionals for the psychological testing and psychosocial therapies.The Center has facilities for routine biochemical and pathological testing. Special investigations like EEG, ECG, X-ray, serum lithium estimation and CAT SCAN etc. are either available at the Center or at SMS Hospital. A Bio-feedback apparatus has been added to the department in the year 1996-1997.There is a separate rehabilitation center for occupational therapy, where patients are given opportunity to work and learn some skills according to their aptitude and therapeutic needs, patients are being involved in the work of carpentry, canning, gardening, painting, in kitchen work, making paper bags for the distribution of medicines in the outpatients department. The rehabilitation therapy department needs to be better equipped to be helpful in the long-term rehabilitation of patients.A regular General Hospital Psychiatric Clinic is being run at S.M.S. Hospital by the consultants and senior postgraduate students from the department. A 20-bedded de-addiction ward has been started at SMS Hospital. It meets the needs of those drug-addicts who do not want to get admitted at the Psychiatric Center. The need for such a facility was long felt.The postgraduate course in psychiatry was started at Jaipur in the year 1979. So far 80 postgraduates have passed out from the department and three new students are registered every year for the M.D. Degree in Psychiatry. Most of the students who have passed out from this center are working in different positions at various places in the state, outside the state in India and abroad.The department is imparting training to both undergraduates and postgraduate student. The training involves theory classes as well as clinical work in outpatient departments. The postgraduate teaching also includes regular case conferences, seminars, tutorials and review of research articles from journals, etc. The postgraduate students also undergo training in Neurology, Neurosurgery, Child Psychiatry, De-addiction Center and Neuroradiology.The institution as a postgraduate and undergraduate teaching department has established itself as one of the important training center in North India. Professors of these departments act as external examiners in about 8-10 universities of the country and this institution has been recognized as an National Board of Examination Center for the National Academy of Medical Science (Dip. NB). Students from all over the country come here for writing the examinations.The students of B.Sc. Nursing come to the center for their practical training in psychiatry. The students of Psychology and General Nursing, from within the state and outside, visit the center for the purpose of teaching and clinical work.The center has also been recognized as one of the five regional center for the implementation of National Mental Health Program (NMHP) of India in the year 1996-1997. Two training programs for the trainers have been organized in the same year, wherein doctors from all the six medical colleges and five district headquarters received the training. Sikar was identified as the district for the implementation of District Mental Health Program under NMHP. Under the training of trainers program supported by Government of India, Psychiatrists from Rajasthan, Punjab, Haryana and Delhi were trained.The department also organizes de-addiction camps in the community with the co-operations of voluntary organization. The center was also given the responsibility of training of doctors from PHC's in de-addiction management.School Mental Health Program has also been initiated by the department and an awareness building program for mental illnesses and drug abuse prevention in school going children has been initiated. A training program for the school teachers was also organized for their orientation and role as counselors in emotional and behavioral problems of school children.There are ample opportunities for research in Psychiatry and related disciplines in behavioral sciences at the center. The postgraduate students through their work on the dissertations are also taught the basic principle of research methodology.The center has been actively involved in psychiatric research. Two projects supported by Indian Council of Medical Research (ICMR), one projects supported by Mental Health Foundation, United Kingdom (Jaipur-London project) apart from the regular departmental research have been completed. The center has also participated in the World Health Organization (WHO) field trials for International Classification of Diseases, putting this department on the world map of Mental Health.Some projects and papers frequently quoted/cited are:-Gehlot PS, Gautam S, Gupta SD. ICMR Epidemiological study of mental illness in a migrant urban population (1981-1983).Gautam S. ICMR Project on Primary Mental Health Care comparative study of efficacy of Training Material and Development of record system (1983-1984).Gautam S, Nijhawan M. Burden on families of schizophrenic, cancer and chronic lung disease patients. Indian J Psychiatry 1984;26:156-9.th Annual Conference of Indian Psychiatric Society at Vishakhapatnam Jan. 1985.Gautam S. Development and Evaluation of Training Programmes for Primary Mental Health care. Indian J Psychiatry 1985;27:51-62. Received “Bhagat Award” for best research paper by scientist under 35 years at 37Gautam S, Nijhawan M, Kamal P. Standardisation of Hindi version of Goldbergs, General Health Questionnaire. Indian J Psychiatry 1987;29:63-6.Gautam S, Nijhawan M. Communicating with cancer patients. Br J Psychiatry 1987;150:760-4: Included in Year Book of Psychiatry and Applied Mental Health 1989.Gautam S. Tilak Venkoba Rao Oration (1989): Post Partum Psy. Syndromes: Are they biologically determied? Indian J Psychiatry 1989;31:31-42.Batra L, Gautam S. Psychiatric morbidity and personality profile in divorce seeking couples. Indian J Psychiatry 1995;37:179-85.Gautam S, Batra L. Sexual behaviour and dysfunction in divorce seeking couples. Indian J Psychiatry 1996;38:109-16 (Marfatia Award Paper of IPS.)Gaita S, Agarwal R, Sharma H. Election: A stressful life event. Indian J Psychiatry 1997;39:329-32.Gautam S, Guptal D, Batra L, Sharma H, Khandelwal R, Pant A. Psychiatric morbidity among victims of bomb blast. Indian J Psychiatry 1999;41:41-5 (Frequently quoted in Disaster Management).Gautam S. IPS Presidential address: Mental Health in ancient India and its relevance to modern psychiatry. Indian J Psychiatry 1999;1:5-18.Apart from this many other research activities are going on in the center. Nearly 200 scientific research papers have been published and presented by the members of the faculty in National and International journals and conferences. Author has received various prestigious awards namely Bhagwat Award, Marfatia Award, Tilak Venkoba Rao Oration Award, Poona Psychiatric Association Award at National Level and International New Horizon Award for work in de-addiction and drug awareness.World Psychiatric Association Symposia and Regional conference 1986, 1998.National Conferences of Indian Psychiatric Society 2003.North Zone Annual Conference of Indian Psychiatric Society 2003.National Mid-term CME of Indian Psychiatric Society (Tradition was initiated from Jaipur) 1990 and 1996.National workshop on ‘Psychiatry Law and Society’ of Indian Psychiatric Society 1989.National Workshop on ‘Clinical Practice Guidelines for Psychiatrists in India’ of Indian Psychiatric Society 2004 and 2005.Training of Trainers Program supported by Government of India organized in 1996, 1997. Trainees included psychiatrists from Rajasthan, Punjab, Haryana and Delhi.Several State Conferences and Workshops of Rajasthan Psychiatric Society have been organized .National Mental Health Program: More than 500 doctors, MPW and community leaders have been trained by this department under National Mental Health Program.Established in 1952 as UG Department of Psychiatry under Department of Medicine, S.M.S. Medical College, Jaipur, the department was running inpatient, Outpatient department and undergraduate medical education up to 1979.Important dignitaries who visited the departmentDr. N.N. Wig, Professor Dmirtus, P.G.I. Chandigarh and Advisor to W.H.O. Regional Center, Alexandria (in 1979 for the inspection of department).Linda B. Johnson, President - American Psychiatric Association (APA) in 1986.Prof. Costa Stefanis,  President, World Psychiatric Association (W.P.A.) in 1986.Dr. J.S. Bajaj, Member Planning Commission - Govt. of India in 1986.Prof. Schulsinger, Secretary General (W.P.A.) in 1986.Prof. Abdul Rasheed Chaudhary, President, Pakistan Psychiatric Association in 1986.Late Justice B.P. Beri, Chairman - Beri Commission and Former Chief Justice of Rajasthan High Court in 1989.Dr. Robert Passnav, President, American Psychiatric Association in 1987.Dr. Melvin Sabshin, Medical Director, American Psychiatric Association in 1986.Prof. G.N.N.R. Reddy, Director  in 1987-1989.Dr. S.M. Channabasavanna, Vice Chancellor (NIMHANS) in 1987Dr. Norman Sartorius, President World Psychiatric Association in 1997 and 1998.Prof. R. Srinivasmurthy, Regional Advisor (WHO) and Former Prof. and Head (NIMHANS), Bangalore (1986-1987) visited in 1998.Prof. Juan Mezzich - USA, Current President (WPA) visited in 1998Prof. Mario Maz (Germany)-President Elect. (WPA) visited in 1998Padamshree Dr. Mohan Agashe-Film Actor and Professor Head Psychiatry, Pune in 1998Prof. H.M. Gadhvi, Senior consultant, London U.K. visiting Professor-1999-2006.Superintendents, HOD and Professors of Psychiatric Center and their TenureDr. B.K. Vyas 24.07.1967 to 31.12.1978Dr. P.S. Gehlot 01.01.1979 to 30.04.1984Dr. J.N. Vyas 28.05.1984 to 03.10.1989Dr. G.B. Advani 04.10.1989 to 18.01.1990Dr. J.N. Vyas 19.01.1990 to 05.05.1990Dr. G.B. Advani 06.05.1990 to 23.02.1996Dr. Shiv Gautam 01.03.1996 to 25.07.2000Dr. L.N. Gupta 25.07.2000 to 19.01.2001.Dr. Shiv Gautam 20.01.2001 to till dateWaiting Hall in the OPD - Psychiatric Center, JaipurPsychiatric Center, Jaipur OPD - Reception, Registration and Drug Dispensing at single windowReception, Psychiatric Center, Jaipur (Front View)OPD Block, Psychiatric Center, Jaipur (Front View)Geriatric Care Center, Psychiatric Center, JaipurDisplay at the Rehabilitation Center, Psychiatric Center, JaipurPaintings made by patientsSome products made by patients at the rehabilitation center


Because of their peculiar sociocultural background, healthcare workers in sub-Saharan African subcultures may have various conceptions on different aspects of autism spectrum disorders (ASD), such as etiology, treatment and issues of prognosis. These various conceptions, if different from current knowledge in literature about ASD, may negatively influence help-seeking behavior of parents of children with ASD who seek advice and information from the healthcare workers. This study assessed the opinions of healthcare workers in Nigeria on aspects of etiology, treatability and preventability of childhood autism, and relates their opinions to the sociodemographic variables.Healthcare workers working in four tertiary healthcare facilities located in the south-east and south-south regions of Nigeria were interviewed with a sociodemographic questionnaire, personal opinion on etiology, treatability and preventability of childhood autism (POETPCA) questionnaire and knowledge about childhood autism among health workers (KCAHW) questionnaire to assess their knowledge and opinions on various aspects of childhood autism.A total of 134 healthcare workers participated in the study. In all, 78 (58.2%), 19 (14.2%) and 36 (26.9%) of the healthcare workers were of the opinion that the etiology of childhood autism can be explained by natural, preternatural and supernatural causes, respectively. One (0.7%) of the healthcare workers was unsure of the explanation of the etiology. Knowledge about childhood autism as measured by scores on the KCAHW questionnaire was the only factor significantly associated with the opinions of the healthcare workers on etiology of childhood autism. In all, 73 (54.5%) and 43 (32.1%), of the healthcare workers subscribed to the opinion that childhood autism is treatable and preventable respectively. Previous involvement with managing children with ASD significantly influenced the opinion of the healthcare workers in subscribing to treatability of childhood autism, while working experience of less than 6 years among the healthcare workers significantly influenced the opinion of the healthcare workers in admitting to believing in the preventability of childhood autism.In designing policies and programs to change negative opinions or beliefs of healthcare workers about childhood autism, there is a need for baseline information such as this survey. Changing the negative opinions or beliefs of the healthcare workers about childhood autism should encourage appropriate help-seeking behavior among parents of children with ASD who may be seeking advice or information from the healthcare workers. This would encourage early interventions, which are essential to prognosis of childhood autism. Adequate and necessary dissemination of information to clients in a healthcare system is part of the essential ingredients of ensuring optimal healthcare performance ,2. InforHelp-seeking behavior and pathway to care are extremely influenced by the opinions or beliefs of the clients and caregivers on the etiology of a particular disorder. Help-seeking behavior and the pathway to care among clients utilizing mental healthcare systems has consistently been noted to be influenced by cultural perspectives and beliefs of clients and caregivers on the etiology of the particular mental disorder in question -6.In the context of sub-Saharan Africa, where healthcare workers living among the general population in the community often play a pivotal role in offering medical advice and providing information on healthcare related issues to people in their immediate community, healthcare workers' cultural perspectives and their opinions or beliefs on etiology of developmental disorders such as ASD would greatly influence the help-seeking behavior of parents of children with ASD living around them in the immediate community.Childhood autism is a developmental disorder that had been observed to have better prognosis with early intervention, which is often achieved through early recognition and diagnosis -9. The nGiven the peculiar subcultures of sub-Saharan Africa including Nigeria, where knowledge and awareness about childhood autism is still relatively low and there exists an imperative need for education of healthcare workers and the public to raise levels of awareness, it is of paramount importance to have some insight into the baseline opinions of healthcare workers in this environment on etiology and issues relating to prognosis and preventability of childhood autism.This study therefore assessed the baseline opinions of healthcare workers working in tertiary healthcare facilities in the south-east and south-south regions of Nigeria on the etiology, treatability and preventability of childhood autism. It also examined the associations between sociodemographic variables of the healthcare workers and their opinions on issues of etiology, treatability and preventability of childhood autism.The locations of the study were four tertiary healthcare facilities located in the south-east and far south regions of Nigeria. Two of these tertiary healthcare facilities are specialized psychiatric hospitals, while the other two are the pediatric departments of two university teaching hospitals. The healthcare facilities where participants were interviewed were University of Calabar Teaching Hospital (UCTH) and Federal Psychiatric Hospital, Calabar, both of which are located in Cross River State (south-south region of Nigeria), and Ebonyi State University Teaching Hospital (EBSUTH), Abakaliki, Ebonyi State and Federal Neuro-Psychiatric Hospital, Enugu, Enugu State, both of which are located in the south-east region of Nigeria.Participating healthcare workers were nurses, either working at the two specialized psychiatry facilities or working in department of pediatrics of the two university teaching hospitals involved in the study. The educational qualifications of the nurses were mostly diplomas in general and psychiatric nursing, and they had been working in their various areas of specialty for at least 1 year. The study was a point survey of opinions of healthcare workers on some aspects of ASD. A point-sampling method that involved all nurses in their duty posts in the four different institutions on the particular day the data were collected was employed. Therefore, all nurses in their duty posts in the four different institutions on that particular day were interviewed.Ethical approval for this study was obtained from the Institutional Review Board (IRB) of Federal Neuro-Psychiatric Hospital, New Haven, Enugu, Enugu State, Nigeria.This was used to obtain sociodemographic information of the healthcare workers such as sex, age, marital status, and area of specialty, among others.This questionnaire was designed to obtain information from healthcare workers about their opinion on etiology, treatability and preventability of childhood autism. The first part of the questionnaire dealt with opinions of the healthcare workers on the etiology of childhood autism. In this part of the questionnaire, healthcare workers were requested to make a choice from a list of four options as to their own opinion on what they thought could be the causal explanation of childhood autism. The four options were: natural causes, preternatural causes, supernatural causes and not sure. For each option they chose, they were further requested to explain or specify what they meant. The second part of the questionnaire dealt with the opinion of the healthcare workers on treatability and preventability of childhood autism. It contained the following questions:• In your own opinion, do you think childhood autism can be treated?• In your own opinion, do you think childhood autism can be prevented?Both questions had the answer options of 'YES' or 'NO', and also further options or space for the healthcare workers to explain or state the reasons for their choice (see Appendix 1).The KCAHW questionnaire measures knowledge about childhood autism aimed at early recognition and diagnosis of ASD among healthcare workers. It is a self-administered questionnaire that contains a total of 19 questions. Each of the questions has three options to choose from, with only one of these three options being correct. The correct option on each question attracts a score of 1, while the other two incorrect options each attract a score of 0. The questionnaire is further divided into four domains. Domain 1 assesses areas of impairment in social interaction and contains eight questions. Domain 2 addresses impairment in areas of communication and language development and contains one question. Domain 3 assesses areas of obsessive and compulsive patterns of behavior found in children with ASD and contains four questions. Domain 4 addresses information on what type of disorder childhood autism is, possible comorbid conditions, and onset of childhood autism in affected children, and contains six questions. Therefore, the KCAHW questionnaire contains a total of 19 questions and possible maximum and minimum total scores of 19 and 0, respectively, when the individual domain scores are added together. The content of the questionnaire and a reliability test for the questionnaire have been described in detail in a previous study . This quThe three questionnaires were distributed to the participating healthcare workers to complete. It was ensured that the questionnaires were completed there and then and collected back immediately from the healthcare workers because they were meant for a point-of-time assessment of opinions on aspects of ASD and knowledge about childhood autism.The data were analyzed using SPSS v.15 . The chi square test was used to determine possible significant associations between sociodemographic variables and opinions of the healthcare workers on issues of etiology, treatability and preventability of childhood autism. The opinions of the healthcare workers on etiological explanation, treatability and preventability of childhood autism were also related with the mean score of the healthcare workers on the KCAHW questionnaire. p Values ≤ 0.05 were considered significant.A total of 134 healthcare workers, which represented the total population on point sampling of the four different institutions studied, consented to participate in the study. There were 71 (53.0%) males and 63 (47.0%) females. The mean age of the participants was 35.89 ± 7.56 years. The mean score of participated healthcare workers on the KCAHW questionnaire was 12.35 ± 4.40. Other sociodemographic variables are shown in Table A total of 78 (58.2%) of the healthcare workers were of the opinion that the etiology of childhood autism is natural, while 36 (26.9%) and 19 (14.2%) subscribed to supernatural and preternatural causes, respectively. One healthcare worker (0.7%) was not sure of the etiology of childhood autism. Those healthcare workers that subscribed to natural causes were likely to attribute the etiology of childhood autism to genetics, birth injury, and maternal infections during pregnancy among other reasons. Those who subscribed to both preternatural and supernatural causes were likely to give explanations such as lineage curses, enemies, and action of the devil, among others.Opinions of the healthcare workers on etiology of childhood autism were only significantly associated with their scores on the KCAHW questionnaire. Healthcare workers having the mean score and above on the KCAHW questionnaire were more likely to subscribe to natural causes for childhood autism compared to those who had scores lower than the mean score on the KCAHW questionnaire  = 3, p value = 0.014). Table A total of 73 (54.5%) of the healthcare workers were of the opinion that childhood autism is treatable, while 43 (32.1%) of the healthcare workers were of the opinion that childhood autism is preventable. The healthcare workers that subscribed to the opinion that childhood autism is treatable cited special education and behavioral therapy among others as possible modalities of treatment. The healthcare workers who were of the opinion that childhood autism is preventable cited avoiding maternal infection during pregnancy, avoiding birth injury, pleasing the ancestral spirit and avoiding sins, among others, as modalities of prevention.A significant association was found between the opinions of healthcare workers on explanation of the etiology and treatability of childhood autism, with those healthcare workers who subscribed to natural causes for childhood autism being more likely to have the opinion that childhood autism is treatable . There was also a significant association between the opinions of healthcare workers on explanation of the etiology and preventability of childhood autism, with those healthcare workers admitting to natural and preternatural causes more likely to subscribe to the opinion that childhood autism is preventable .Opinions of healthcare workers on the treatability of childhood autism showed significant association with being previously involved with management of children with ASD, with those healthcare workers who have had previous involvement in managing children with ASD more likely to subscribe to the opinion that childhood autism is treatable when compared to those who had not been involved before in the management of children with ASD . Opinions of the healthcare workers on treatability of childhood autism showed near-significant association with geographical region, with the healthcare workers located in the south-east region of Nigeria more likely to express the opinion that childhood autism is treatable compared to those located in the south-south region of the country . Near-significant association was also found between opinions of the healthcare workers on treatability of childhood autism and their scores on the KCAHW questionnaire, with those healthcare workers having the mean score and above on the KCAHW questionnaire more likely to subscribe to the opinion that childhood autism is treatable when compared to those who had scores lower than the mean score on the KCAHW questionnaire . Table A significant association was found between the opinions of healthcare workers on the preventability of childhood autism and years of working experience, with those healthcare workers with working experience below 6 years more likely to express the opinion that childhood autism is preventable . Near-significant association was found between opinions of the healthcare workers on preventability of childhood autism and gender, with females more likely to subscribe to the opinion that childhood autism is preventable compared to males . Near-significant association was also found between opinions of the healthcare workers on the preventability of childhood autism and age group, with those healthcare workers below 40 years of age more likely to express the opinion that childhood autism is preventable . Table Childhood autism is a pervasive developmental disorder, where a definitive etiology is still obscured. What is certain to date is that it occurs as a result of developmental problems in the brain that affect phenotypic areas of communication and social interaction, and it brings about restricted areas of interest or repertoire in the affected child . A bioloAnother interesting finding of this study was that the more apt the healthcare workers were at being able to recognize the symptoms of childhood autism (as reflected by their scores on the KCAHW questionnaire), the more likely they were to express an opinion of natural causes as explanation for the etiology of childhood autism. Promoting knowledge among healthcare workers about childhood autism through continuing education and special training would not only help in early recognition and diagnosis, but also positively influence the help-seeking behavior of parents of children with ASD who would come into contact with these healthcare workers.Current knowledge advocates a multidisciplinary approach to the management of childhood autism. This management approach incorporates special education, behavioral therapy, social and communication skill training and psychotropic medications, if indicated . With eaAbout 55% of the healthcare workers that participated in this study were of the opinion that childhood autism is treatable. This gave us some hope in the sense that these healthcare workers would be able to reassure parents of children with ASD and give appropriate information that would be necessary to their seeking help. However, what is lacking in most third-world regions such as Nigeria is a multidisciplinary approach to the management of children with developmental disabilities in general, largely due to the absence of trained professionals and lack of facilities for special needs of children with developmental disorders.About 32% of the healthcare workers subscribed to the opinion that childhood autism is preventable. The premise for this position among the healthcare workers was often based on the theory of insult to the developing brain, either as a result of maternal infection, intrauterine infection or physical trauma to the brain from delivery or birth complications, and also the theory of supernatural causes as explanation for the etiology of childhood autism, which rest on (lack of) forgiveness of sins and trespasses, and cutting off links with cursed ancestral spirits among others. Current knowledge however has not afforded the scientific community the opportunity of knowing the definitive etiology of ASD and therefore definitive steps that are needed to ensure prevention also remained obscured.Significant associations existed between the healthcare workers' opinions on the etiology, treatability and preventability of childhood autism. Essentially, the opinions of the healthcare workers that participated in the study on the etiology of childhood autism had significant influence on their opinions about whether childhood autism is treatable and/or preventable. This finding is in line with the general phenomenon that the etiology of any disorder often influences its treatment and preventability.The opinions of the healthcare workers on whether childhood autism is treatable or can be managed were significantly influenced by previous involvement with managing children with ASD. Those healthcare workers who had been involved in the management of children with childhood autism were more likely to show optimism that the condition can be managed or treated.The opinions of the healthcare workers on whether childhood autism is preventable were significantly influenced by number of years of working experience of the healthcare workers, with those who had less than 6 years work experience more likely to subscribe to the opinion that childhood autism is preventable. The reason for this finding is not clear, but it could be due to the limited experience of these healthcare workers in working with children with developmental disorders.The significance of this baseline study is in assessing the areas where attention needs to be focused among healthcare workers in changing their negative attitudes and beliefs on various aspects of childhood autism; a process that is envisaged as necessary in promoting care for children with ASD and other developmental disorders in this environment. More attention therefore needs to be focused on specialized training on ASD for healthcare workers working with children and adolescents in this environment.The present information on the opinions of the participating healthcare workers on various aspects of childhood autism serve a baseline on which future policies and programs to change negative opinions and beliefs of healthcare workers in this environment on various aspects of childhood autism can be based.Changing the negative opinions or beliefs of the healthcare workers about childhood autism would encourage appropriate help-seeking behavior among parents of children with ASD, who would be seeking advice or information from the healthcare workers. This would encourage early intervention, which is essential to the favorable prognosis of childhood autism.The authors declare that they have no competing interests.All authors contributed to the conception of the study. MOB was involved with writing the initial draft of the manuscript. MOB, AOA, POE, JE, KOO, JUO and GMO were involved in revising the manuscript. All authors read and approved the final draft of the manuscript.Kindly answer the following questions to the best of your opinion. Please do not consult formal textbooks to answer these questions. Thank you for your time.1.   In your own opinion, what is the likely causal explanation of childhood autism among the possible causal explanations listed below (tick one and give reasons for your choice)a.   Natural causes (specify)...............................................................b.   Preternatural causes (specify).........................................................c.   Supernatural causes (specify).........................................................d.   Not sure (specify).......................................................................2.   In the questions below tick one appropriate option to the best of your opinion:i.   In your own opinion, do you think childhood autism can be treated?   (a) YES   (b) NO (specify why).................................................................ii.   In your own opinion, do you think childhood autism can be prevented?   (a) YES   (b) NO (specify why)...................................................................


The Drug Abuse Resistance Education (D.A.R.E.) program is a widespread but controversial school-based drug prevention program in the United States as well as in many other countries. The present multivariate meta-analysis reviewed 20 studies that assessed the effectiveness of the D.A.R.E. program in the United States. The results showed that the effects of the D.A.R.E. program on drug use did not vary across the studies with a less than small overall effect while the effects on psychosocial behavior varied with still a less than small overall effect. In addition, the characteristics of the studies significantly explained the variation of the heterogeneous effects on psychosocial behavior, which provides empirical evidence for improving the school-based drug prevention program. Drug abuse is a prevalent problem affecting young generations worldwide . In respdrug use, which includes tobacco, alcohol, marijuana, and other illicit drugs; and (b) the improvement of psychosocial behavior, which includes social skills , self-esteem, attitudes towards drug use, attitudes towards police, and family bonding. The program is normally taught by a police officer; and the core curriculum has 17 lessons, usually offered once a week for 45 to 60 minutes [et al. [The D.A.R.E. program was designed to help elementary and junior high school students resist the peer-pressure of experimenting with drugs, tobacco, and alcohol. The D.A.R.E. program aims to reduce drug abuse among children by providing them with information that encourages them to make healthy decisions. Its effectiveness has been assessed by its two major outcomes: (a) the reduction of  minutes . This tyAlthough there seems to be great support for the D.A.R.E. program, the literature revealed inconsistent research results concerning the program’s effectiveness. For example, some studies found that the D.A.R.E. program did not work , 8–15; wet al. [Considering the tremendous investment of time and money in the D.A.R.E. program, these inconsistent findings necessitate a conclusive synthesis of the research to assess the effectiveness of the program. To date, only two published research syntheses or meta-analytic reviews exist that solely focused on evaluating the effectiveness of the D.A.R.E. program , 10. Unfet al. , 37. Weset al. , on the et al. . Becauseet al.  serves aThe purpose of this multivariate meta-analytic review was to: (a) quantitatively synthesize updated evaluation studies of the D.A.R.E. program, and (b) simultaneously synthesize all the outcomes of the D.A.R.E. program. Specifically, this review addressed the following three research questions: (a) Did the effects of the D.A.R.E. program on the outcomes vary across the studies? (b) What was the overall effect of the D.A.R.E. program on the outcomes? (c) What study characteristics explained the variation of the effects of the D.A.R.E. program on the outcomes?Using the terms “Drug Abuse Resistance Education,” “D.A.R.E.,” and “school-based drug prevention program” as keywords, an extensive literature search in the PsycINFO, MEDLINE, ERIC, PUBMED, SOCIAL SCIENCE INDEX, SOCIAL SCIENCE INDEX CITATION, and DIGITAL DISSERATIONS databases produced 198 relevant studies published between January 1983 and December 2005. Of the 198 studies, 73 were found to be quantitative studies on the D.A.R.E. program. After applying the criteria for inclusion described below, 20 final studies were selected for this meta-analysis.d [d is a standardized mean difference between the treatment group and the control group. That is,x̄t is the mean of the treatment group, x̄c is the mean of the control group, and sp is the pooled standard deviation. Whether the study utilized an experimental or quasi-experimental design was the second criterion because these designs are more rigorous and provide more valid research results than other less scientific designs. The third criterion necessitated that the study evaluated at least one of the outcomes on drug use and psychosocial behavior. The fourth and final criterion called for studies where the effect of the D.A.R.E. program could be independently evaluated. That is, whether the studies provided a D.A.R.E. treatment group and a comparable control group.The first criterion for inclusion required the study to have sufficient quantitative information for calculating the outcome measure or the effect size of the outcome: Cohen’s d . Cohen’sOutcome measures. The outcome measures for the present review were two sets of effect sizes: one was for drug use and the other for psychosocial behavior. Each effect size in the former set was the average Cohen’s d for all the available drug use outcomes , and the latter for all the available psychosocial behavior outcomes . Different effect size measures calculated from various statistical methods in the studies were converted to Cohen’s d. In line with the previous reviews [ds of the outcomes across the studies were calculated at the longest follow-up, which ranged from 0  to 10 years. reviews , 10, theStudy characteristics. The following study characteristics were recorded for the analysis: Name of first author, year of publication, sample size, statistical method , year of D.A.R.E. curriculum, follow-up time, proportion of female participants, and proportions of ethnic groups. The selection of the study characteristics was partially guided by the pervious reviews and partially based on common information available in the studies.Each value of the variables of the study characteristics and outcome measures needed to be recorded or coded from the 20 studies. A concurrent double coding was performed independently by the researchers. Each researcher spent more than forty hours, equivalent to five full-time work days, on coding the 20 studies. Then, the researchers engaged in extensive discussions to compare every coded item. No variable was finalized until reaching an agreement.d = 0.20, 0.50, and 0.80 are considered small, medium, and large effect, respectively. 95% confidence intervals of the effect sizes were also computed. The confidence intervals showed whether the effect sizes were heterogeneous across the studies.Descriptive analysis was first conducted for each of the two outcomes, drug use and psychosocial behavior, by calculating the unweighted mean effect size of the outcome. According to Cohen’s guideline , d = 0.2Q-statistic was computed for each of the two outcomes, drug use and psychosocial behavior. The test for the Q-statistic provided statistical evidence for the heterogeneity of the 20 studies. If the test was significant, a random-effects model was tested, and the weighted mean effect size was calculated to provide a more valid estimate for the mean effect size than the unweighted mean effect size from the descriptive analysis.In terms of inferential analysis, a Hedges and Olkin’s  Q-statisweighted regression model to explain the variation in the heterogeneous effect sizes. Following Hedges’ [t-test for individual regression coefficient was adjusted as follows:s.e. is the original standard error given by common computer programs, and MSError is the mean square value for errors from the analysis of variance for the regression given by the computer programs. Note that some study characteristics had missing values that resulted from unavailable information, and they were replaced by means [In the case of heterogeneous effect sizes, the study characteristics were entered into a  Hedges’  suggestiby means  because by means –45.Test of homogeneity. Under the null hypothesis of H0: θ1 = … = θ20 = θ, the Hedges and Olkin’s [Q-statistic values of QTotal were 13.34 with df = 17 (p = 0.71) and 96.61 with df = 12 (p < 0.0001) for drug use and psychosocial behavior, respectively. The homogeneity test results showed that the effect sizes across the 20 studies were statistically heterogeneous for psychosocial behavior, but not for drug use. This inferential finding was consistent with the descriptive finding demonstrated in the confidence interval plots above. By testing a random-effects model for psychosocial behavior under H0: θ = 0, a z = 2.92 (p < 0.01) indicated that the weighted average effect size of the 20 studies from the random-effects model was statistically different from zero but was still 0.10, a less than small effect. Olkin’s  Q-statisWeighted regression analysis. Because the effect sizes were heterogeneous for psychosocial behavior, a weighted regression analysis was conducted to identify the study characteristics that explained the heterogeneity. R2 = 89.8%). Specifically, the longer follow-up time  and the more rigorous statistical method  the study used, the less effect of the D.A.R.E. program would be found for psychosocial behavior; whereas the later D.A.R.E. year , the more White students , and the more Black students  the study had, the more effect of the D.A.R.E. program would have on psychosocial behavior.By including more updated studies and analyzing the study characteristics related to the outcomes of the D.A.R.E. program on both drug use and psychosocial behavior, this present multivariate meta-analysis provided a more comprehensive review than previous ones on the effectiveness of the D.A.R.E. program; and therefore, the present review helps us to better understand the widespread, expensive, but controversial D.A.R.E. program. The results of the present review revealed that the effects of the D.A.R.E. program on drug use were homogeneous but less than small, which confirmed the findings in the literature –11, 13. For the heterogeneous effects of the D.A.R.E. program on psychosocial behavior, the present review found that the study characteristics explained most of the variation of the effects. The heterogeneous effects suggest that some studies showed larger effects than others. By examining the specific characteristics of the studies that had larger effects, which was executed in the weighted regression analysis, future program implementations can learn from those effective studies for improving the program effects. Among the significant study characteristics, follow-up time and statistical method were negatively related to the effects; and D.A.R.E. year, percent of White participants, and percent of Black participants positively related to the effects.et al. [These findings provided some important implications. First, the validity of long-term effects might be threatened by maturity and history. This point was also noted in the previous reviews , 10 but et al. . The newet al. . Some otet al. . As suchd = 0.05); on the other hand, the school-based drug intervention program also had a less than small effect on improving psychosocial behavior (Cohen’s d = 0.10). The analysis from this review also identified areas in the new versions of the D.A.R.E. program that need improvement. It would be, however, more important if the new versions of the D.A.R.E. program could transform the improved psychosocial behavior into the students’ actions of reducing drug use—the ultimate outcome.In sum, the effects of the D.A.R.E. program appear to be different on drug use and psychosocial behavior. The results of the present review provide an evidence-based interpretation to the inconsistent conclusions found in the previous research that was conducted on the D.A.R.E. program. This study found that, on one hand, the D.A.R.E. program had a less than small effect on reducing drug use (Cohen’s


Anopheles lesteri, Anopheles sinensis and Anopheles pullus, have long been suspected malaria vectors. However, opinions about their vector ability are controversial. The present study was designed with the aim of determining the susceptibility of these mosquitoes to a Korean isolate of Plasmodium vivax. Also, An. sinensis is primarily suspected to be vector of malaria in Korea, but in Thailand, the same species is described to have less medical importance. Therefore, comparative susceptibility of Thai and Korean strains of An. sinensis with Thai strain of P. vivax may be helpful to understand whether these geographically different strains exhibit differences in their susceptibility or not.Following its recent re-emergence, malaria has gained renewed attention as a serious infectious disease in Korea. Three species of the Hyrcanusgroup, An. lesteri, An. sinensis and An. pullus was studied by feeding laboratory-reared mosquitoes on blood from patients carrying gametocytes from Korea and Thailand.The comparative susceptibility of P. vivax, oocysts developed in An. lesteri, An. sinensis and An. pullus. Salivary gland sporozoites were detected only in An. lesteri and An. sinensis but not in An. pullus. Large differences were found in the number of sporozoites in the salivary glands, with An. lesteri carrying much higher densities, up to 2,105 sporozoites in a single microscope field of 750 × 560 μM, whereas a maximum of 14 sporozoites were found in any individual salivary gland of An. sinensis. Similar results were obtained from a susceptibility test of two different strains of An. sinensis to Thai isolate of P. vivax, and differences in vector susceptibility according to geographical variation were not detected.In experimental feeding with Korean strain of An. lesteri indicate that this species is highly susceptible to infection with P. vivax. Anopheles sinensis appears to have a markedly reduced ability to develop salivary gland infection, whilst in An. pullus, no sporozoites were found in the salivary glands. Provided that the survival rate of An. lesteri is sufficiently high in the field, it would be a highly competent vector of vivax malaria.The high sporozoite rate and sporozoite loads of  Plasmodium vivax malaria has re-emerged in many malaria-endemic areas, where the disease was believed to have been eradicated [adicated . In Soutadicated -5. In thadicated . As a reAnopheles sinensis, Anopheles lesteri, Anopheles pullus, Anopheles sineroides, Anopheles kleini and Anopheles belenrae [Anopheles koreicus and Anopheles lindesayi japonicus [So far, eight anopheline mosquitoes have been reported from South Korea. Six of these species belong to the Hyrcanus group: belenrae -10. Two aponicus .An. sineroides, because these species are very rare in Korea, Japan and China and, are thus considered medically less important [An. belenrae and An. kleini [An. lesteri, An. sinensis and An. pullus are controversial. Anopheles sinensis has long been considered a primary and strong vector in Korea and Japan [Since vivax malaria has been recognized as an endemic disease particular to countries in the Far East Asia, including Korea, efforts to find the main vector(s) have continuously proceeded from Korea, Japan and China ,11-13. Hmportant ,14,15. A. kleini , the bit. kleini . The resnd Japan ,15,17, wnd Japan ,19.An. lesteri has been considered a minor and weak vector in Korea and Japan, studies in China showed that it was an important vector [An. pullus may also be found [Although t vector . Conflicbe found ,4, with be found . Thus, tAn. lesteri, An. sinensis and An. pullus were experimentally infected with a Korean isolate of P. vivax and the malarial susceptibility of these species was analysed based on their ability to develop oocysts in the midgut and sporozoites in salivary glands. To substantiate the findings of the above experiments and to further understand and verify the existing differences in vector abilities of two geographically distant strains of An. sinensis from Korea and Thailand, experimental infections were conducted with a Thai isolate, and the ability of both these strains to develop oocysts and sporozoites was determined.In this study, three species of the Hyrcanus group, Anopheles raised in the laboratory were An. lesteri, An. sinensis and An. pullus. Among these, An. sinensis and An. pullus were collected from Paju City, Gyeonggi-do Province, while An. lesteri was collected from So-Rae District, Incheon City in South Korea. The Thai strain of An. sinensis was collected from Mae Tang District, Chiang Mai Province, Thailand. Anopheles cracens was in a free-mating colony established for more than two decades in the insectariums of the Department of Parasitology, Chiang Mai University, Thailand and was used as a control, since it is known to be highly susceptible to P. vivax [et al [Three mosquito lines of the Korean P. vivax . These cx [et al  in an inP. vivax was supplied by the National Institute of Health (KNIH), from a patient seeking treatment at Gimpo, Gyeonggi-do province. The Thai strain of P. vivax was obtained from malaria patient infected in Mae Tang District of Chiang Mai Province. Informed consent was obtained from the patients before collection and the study protocols were approved by Internal Review Board of Korea National Institute of Health and Thailand Office for Vector Borne Diseases Control, Department of Communicable Diseases Control, Ministry of Public Health. Giemsa staining of the blood film was performed and gametocytes were counted. The gametocyte density of the Korean isolate of P. vivax was 36 gametocytes/200 White Blood Cells (WBC) while that of the Thai strain was 26 gametocytes/200 WBC.Blood containing gametocytes of Korean et al [Three to five days old females were used for infection. To enhance their willingness to feed, the mosquitoes were starved for 12 hours prior to infection and were transferred to paper cups of size 8.5 cm in diameter and 11 cm in depth . The mosquitoes were fed on infected blood containing gametocytes through an artificial membrane feeding technique described by Chomcharn et al . The mosAn. lesteri, An. sinensis and An. pullus were infected with Korean isolate of P. vivax, while strains of An. sinensis from Thailand and Korea together with An. cracens were infected with Thai isolate of P. vivax.Unfed mosquitoes were removed and the fully engorged females were carefully handled and were kept in the insectary. Mosquitoes were then fed on cotton patches, which were soaked in 5% sucrose solution and changed every day, until the mosquitoes were dissected. During infection, three lines of the Korean mosquito: Eight and 14 days after feeding, mosquitoes were dissected to detect oocysts in the midgut and sporozoites in the salivary glands respectively. Counting of oocysts in mosquitoes was followed by examining wet mounts of the midgut stained in 0.1% mercurochrome and freely moving sporozoites were carefully detected from salivary glands placed in a drop of phosphate buffered saline .An. lesteri and An. sinensis, two methods were used: 1) for An. lesteri, micrographs were taken at 100× magnification using a camera (Motic Cam 2000) mounted on a compound microscope (Leica DM 2500) and counting of sporozoites was performed within a captured micrograph, the corresponding area of which was approximately 750 μm × 560 μm; 2) sporozoites were counted from whole salivary glands from An. sinensis and were compared to the sporozoite loads within a single microscopic field of An. lesteri. The above counting and comparative procedures were developed because in highly infected An. lesteri, the direct counting of several hundreds of moving sporozoites while observing through the microscope was difficult. It was sometimes impossible to count all of them from salivary glands while in An. sinensis any such possibilities were less and few numbers of sporozoites were directly counted by changing fields. Depending on counting, mosquitoes were graded as 1+ (1–10 sporozoites), 2+ (11–100 sporozoites), 3+  and 4+  [To explore differences in densities of the salivary gland sporozoites between Leica DM 00 and coP. vivax. However, An. lesteri was highly susceptible, in that all nine mosquitoes carried oocysts (100%). Compared to An. lesteri, fewer percent of An. sinesis (87%) and An. pullus (83%) developed oocysts.As indicated by oocysts developed in the midgut, infections were detected in all three species eight days post feeding with Korean strain of An. lesteri contained oocysts. However, fewer An. sinensis and An. pullus do so. In addition, when detached salivary glands of the infected mosquitoes were examined, nine out of 14 An. lesteri (64%) and only two out of 19 An. sinensis (11%) contained sporozoites, while the examined salivary glands of all 6 An. pullus lacked sporozoites  and the other had 2+ (11–100 sporozoites). While the lowest and highest numbers of sporozoites counted in one microscopic field of An. lesteri were 78 and 2,105. Among 9 infected An. lesteri, one had gland index of 2+ (11–100 sporozoites), four were with 3+  and remaining four were with 4+ , respectively. When compared to An. sinensis, the lowest and highest numbers of sporozoites of An. lesteri, even in a single microscopic field, were 5–150 times more intense.Even if few numbers of w Figure . The higP. vivax, equal percent of An. sinensis from Thailand and Korea (88%) develop oocysts, except differences were seen in number of the midgut oocysts they carried. All seven dissected specimens of An. cracens (100%), the control species, produced higher number of oocysts compared to both strains of An. sinensis.When mosquitoes were dissected eight days after feeding with Thai strain of An. sinensis while only one out of 20 specimens of Korean strain (5%) contained salivary gland sporozoites. Furthermore, salivary gland sporozoites were detected from of all An. cracens (100%), the control species  and also pointed out that An. sinensis, due to its zoophilic behaviour, is an inefficient vector responsible for maintaining a low malaria endemicity in the broad flat rice plains of south China. Similarly, Harrison [An. lesteri might well be a more efficient vector than An. sinensis. Lately, Liu [Anopheles anthropophagus (synonymized as An. lesteri) [P. falciparum gametocytes and also reported the natural infection rates of An. lesteri to be higher than An. sinensis in China. With such findings and along with other essential parameters , An. lesteri was considered to be 20 times more efficient as a malaria vector than An. sinensis in malaria infected regions in China [An. sinensis were reported in 1962 [An. sinensis followed by An. pullus . These early studies supported the primary and secondary vector status of An. sinensis and An. pullus.Ho et al  showed tHarrison  opined tely, Liu  recordedlesteri)  from expin China . In Sout in 1962 . In a si in 1962  also repAnopheles was based on Enzyme-Linked Immunosorbent Assay (ELISA). Most of these studies indicated An. sinensis to be a vector [et al [An. kleini and An. pullus to be stronger vectors than An. sinensis. Because Circumsporozoite Protein (CSP) detected by ELISA can equally be detected from developing oocysts, and sporozoites present in haemocoel, positive results from the test do not always indicate that the salivary glands are infected with sporozoites [In recent years, much of the understanding of vector susceptibilities of Korean a vector ,29,30, wr [et al  reportedrozoites . Thus, iAn. lesteri, An. sinensis and An. pullus) were experimentally infected with an indigenous Korean isolate of P. vivax. After eight and 14 days post-infection, examination of midguts showed the presence of high number of oocysts in all three species. However, there were differences in their innate ability to develop sporozoites in the salivary glands. Sporozoites in salivary glands were detected from An. lesteri and An. sinensis but not from An. pullus. Though, sporozoites were detected from An. sinensis, they were very few as compared to An. lesteri. The maximum number of sporozoites in a salivary gland of An. sinensis was 14, which corresponded to the findings of Rongsriyam et al [et al [An. sinensis can be assumed to play less important role in initiating infections when compared to An. lesteri.In this study, three Korean species of Hyrcanus group (am et al . Assuminl [et al  reportedP. vivax at 25°C is completed within 9 days [An. lesteri, very few number of An. sinensis contained salivary gland sporozoites however no sporozoites were detected in An. pullus. This suggests that developmental transitions couldn't proceed from oocysts to sporozoites formation in An. sinensis and An. pullus. Beier et al [Because sporogony in n 9 days , fourteeer et al  describeAn. lesteri. Therefore, future studies regarding survival rate of An. lesteri in wild population can be more supporting evidence for this study. In Korea, the present study is the first of its kind, comparing the malarial susceptibilities of these members through successful malaria development within lab-raised clean colonies. The results are well congruent with Chinese and Thailand reports. So, the outcomes of this study have significant bearings within entire temperate regions where these species are abundant.In addition to demonstrating sporozoites in salivary glands following laboratory infection, it is necessary to consider the natural survival rates of malaria vectors. At present, there are no connected reports with the matter in An. lesteri, as readily as in the well-recognized vector. Big differences were seen in the rate and densities of sporozoites between An. lesteri and An. sinensis, whereas sporozoites were not detected from salivary glands of An. pullus even after 14 days of oocysts development. Also, geographically distant strains of An. sinensis from Korea and Thailand were similar in their ability to support malaria development. Therefore, An. lesteri is highly susceptible to P. vivax malaria as compared to An. sinensis and An. pullus.Under laboratory conditions, salivary gland sporozoites were developed in An. lesteri is a potential malaria vector and its presence may be described as an under-rated public health threat. However, comparative susceptibility of the remaining members of the Hyrcanus group will be important in future to understand their role in malaria epidemiology in Korea.Thus, The authors declare that they have no competing interests.Anopheles mosquitoes. DJ and MHP conducted all experimental studies under the supervision of MGS and WC. Malaria-infected blood was obtained from WC, WS, TSK and JYK. DJ drafted the manuscript with MHP. All the authors read the manuscript.MGS designed the present study. Prior to this experimental work, WC facilitated DJ and MHP for training on malaria susceptibilities and improved lab colonization of


Anopheles gambiae and Anopheles funestus mosquito species complexes are the primary vectors of Plasmodium falciparum malaria in sub-Saharan Africa. To better understand the environmental factors influencing these species, the abundance, distribution and transmission data from a south-eastern Kenyan study were retrospectively analysed, and the climate, vegetation and elevation data in key locations compared.The An. gambiae sensu strict, Anopheles arabiensis and An. funestus entomological inoculation rates (EIRs), were used as focal points for spatial and environmental analyses. Transmission patterns were examined for spatial autocorrelation using the Moran's I statistic, and for the clustering of high or low EIR values using the Getis-Ord Gi* statistic. Environmental data were derived from remote-sensed satellite sources of precipitation, temperature, specific humidity, Normalized Difference Vegetation Index (NDVI), and elevation. The relationship between transmission and environmental measures was examined using bivariate correlations, and by comparing environmental means between locations of high and low clustering using the Mann-Whitney U test.Thirty villages in Malindi, Kilifi and Kwale Districts with data on An. arabiensis and An. funestus transmission, but not of An. gambiae s.s., which was found to be widespread across the study region. The spatial clustering of high EIR values for An. arabiensis was confined to the lowland areas of Malindi, and for An. funestus to the southern districts of Kilifi and Kwale. Overall, An. gambiae s.s. and An. arabiensis had similar spatial and environmental trends, with higher transmission associated with higher precipitation, but lower temperature, humidity and NDVI measures than those locations with lower transmission by these species and/or in locations where transmission by An. funestus was high. Statistical comparisons indicated that precipitation and temperatures were significantly different between the An. arabiensis and An. funestus high and low transmission locations.Spatial analyses indicated positive autocorrelation of These finding suggest that the abundance, distribution and malaria transmission of different malaria vectors are driven by different environmental factors. A better understanding of the specific ecological parameters of each malaria mosquito species will help define their current distributions, and how they may currently and prospectively be affected by climate change, interventions and other factors. Plasmodium falciparum malaria is primarily transmitted by mosquito species belonging to the Anopheles gambiae and Anopheles funestus complexes [An. gambiae sensu lato and An. funestus are present, as they tend to exploit different breeding habitats and peak at different times, thereby prolonging the transmission period. Generally, Anopheles gambiae s.l. are most abundant during the rainy season, and An. funestus is predominant at the end of the rains and beginning of the dry season [et al [An. gambiae s.l. and An. funestus, and compare climate, vegetation and elevation data derived from remote-sensed satellite sources in key locations.In sub-Saharan Africa, omplexes -4. The iomplexes . High ley season -3. The en [et al , provideet al [An. gambiae s.l. and An. funestus at 30 villages in the Malindi, Kilifi and Kwale Districts along the south-eastern coast of Kenya. Mosquito collections between June 1997 and May 1998 indicated that An. gambiae sensu stricto, Anopheles arabiensis and An. funestus were the main malaria vectors, with differing geographical abundance and transmission patterns over the 200 km study area. Interestingly, An. gambiae s.s. was found to be widespread, whereas An. arabiensis was mostly confined to Malindi in the north and An. funestus to Kwale in the south. Preliminary climate analyses by Mbogo et al [An. gambiae s.l. and An. funestus, however, these varied by species and between districts, and climate data were limited to one meteorological station in each district.The study by Mbogo et al  providesgo et al , found pet al [An. gambiae s.s., An. arabiensis and An. funestus clustered locations were carried out.The recent advances in space technology and increased public access to remote-sensed satellite data provide a cost-effective and efficient alternative to examine relationships between climate, the environment and mosquito vectors of human disease ,8. This et al  study, cAn. gambiae s.l and An. funestus . Third, the spatial patterns of An. gambiae s.s, An. arabiensis and An. funestus transmission were examined in ArcGIS using Spatial Analyst tools . The Moran's I statistic was used to determine spatial autocorrelation patterns i.e. clustered, dispersed, random, and the Getis-Ord Gi* statistic was to identify the locations with high and low clustering  -1.96 and +1.96 standard deviations). The distributions of clustering across the study area were highlighted in relation to elevation, using a 3D wireframe map created in the surface mapping programme Surfer 7.0 . Mean environmental measures between high and low clustering trends were compared using the Mann-Whitney U test with Bonferroni correction for multiple comparisons. All statistical analyses were performed in Microsoft Excel and SPSS 16.0 .First, the average number and daily EIRs of le et al ), and clle et al ), and eaClimate and vegetation data corresponding to the 30 mosquito collection sites (i.e. latitude and longitude), and original time period i.e 1997-1998) were obtained from the best available sources, accessed via the IRI/LDEO Climate Data Library of the International Research Institute for Climate and Society 97-1998 w-12. VegeAn. gambiae s.l., had significantly (95% CI) higher precipitation, but lower temperature, specific humidity, NDVI and elevation measures than the 10 sites from Kwale District in the south, where An. funestus was most prevalent. Overall, these trends are supported by the correlations between the three main species, and each environmental variable  and An. arabiensis (r = -0.385) were negatively correlated with An. funestus, which is in accordance with observations by Mbogo et al [The findings of these analyses suggest that the different mosquito species compositions found in Malindi, Kilifi and Kwale Districts during 1997 and 1998 may be related to their different climate and topographical profiles. Figure go et al .An. arabiensis  and An. funestus , but not for An. gambiae s.s. . The resultant Z scores of the Getis-Ord Gi* hot spot analyses (using inverse-distance weighting), indicated similar trends with significant spatial clusters of high and low EIR values found for An. arabiensis and An. funestus but not for An. gambiae s.s. The clustering trends are shown in Figure An. gambiae s.s, 17 locations had positive Z scores (ranging 0.38 to 1.73) predominantly in the north, while the remaining 13 locations had negative Z scores (ranging -0.16 to -1.70) predominately in the south. For An. arabiensis, six locations with high EIR values were significantly clustered (Z scores ≥ 1.96) in Malindi District, and two with low EIR values (Z score ≤ -1.96) in Kwale District. This contrasts to An. funestus, which had five high EIR values significantly clustered in Kwale District, and five with low EIR values in Malindi District.Spatial analyses indicated positive spatial autocorrelation or clustering for An. gambiae s.s and An. arabiensis showed similar environmental trends, with locations with higher transmission having higher precipitation, but lower temperature, humidity and NDVI measures than those locations with lower transmission by these species and/or where transmission by An. funestus was higher. Notably, locations with higher An. arabiensis transmission trends had markedly low elevations, also illustrated in Figure An. gambiae s.s there were no significant differences , while for An. arabiensis precipitation and temperatures were found to be significantly different, and for An. funestus precipitation, temperatures and humidity were found to be significantly different between the higher and lower transmission locations.For each species, comparisons of environmental measures between locations with high and low transmission trends are shown in Table An. gambiae s.l and An. funestus can have distinct ecological niches and requirements within a relatively small geographical area. This is supported by other entomological studies carried out in the region, which highlight the heterogeneous nature of these species' seasonality, host feeding preferences [An. gambiae s.s larvae mostly occur in open shallow sunlit puddles and pools close to homesteads, whereas An. funestus larvae prevail in permanent vegetated aquatic habitats such as stream pools of rivers. In general, malaria transmission by An. funestus predominantly occurs in rural areas of sub-Saharan Africa, and the fact that Kwale District was less urbanized than the other districts [An. funestus prevailed in this region. Furthermore, the presence of both An. gambiae s.s. and An. funestus, whose ecological requirements may be complementary to each other [These simple comparative analyses of 30 sites across three districts in Kenya indicate that ferences -16, bodyferences  and the ferences -20. For istricts ,22, may ch other , may alsAn. funestus but not by An. arabiensis [An. funestus than for An. gambiae [Changes in the local environment are important to understand because they can create, or reduce the number of, suitable breeding sites for local vectors, thereby affecting their abundance and transmission patterns. In central Kenya, the introduction of irrigated rice cultivation appeared to reduce the risk of malaria transmission by abiensis , and in  gambiae . Along t gambiae , agricul gambiae ,27, whicAnopheles species, and a change in mosquito composition and biting times of An. gambiae s.l. [An. funestus with a shift among the outdoor resting females from endophagy on humans to exophagy on animals [An. funestus can be readily eliminated from an entire area by IRS programmes. However, this vector can reappear and become widespread again, sometimes with resistance to the insecticides used in the spray campaign [Similarly, the impact of interventions such as insecticide-treated bed nets (ITNs), long-lasting insecticide-treated nets (LLINs) and indoor residual spraying (IRS) should be considered as they may affect species differently, especially if distributed widely over large geographical areas. The introduction of ITNs in Kilifi and Kwale District during the 1990s significantly reduced the number of indoor-resting iae s.l. , and in  animals . However animals ,30 and e animals -33 have campaign -36. Thiscampaign , especiacampaign .An. gambiae s.s. and An. arabiensis, An. gambiae s.s had no significant clustering or environmental differences between high and low transmission locations. The reasons for this may be related to its wide distribution and ability to exploit a range of habitats [An. gambiae s.s have shown to have differing spatial distributions and environmental parameters [An. gambiae S form has been detected in two locations [An. gambiae s.s forms is crucial, especially as An. gambiae s.l appears to be the main vector of both malaria and lymphatic filariasis in Kilifi and Kwale Districts [Although there was considerable overlap between habitats -3,18, burameters -41, and rameters . In thisocations , therefoistricts ,44,45.et al [An. arabiensis, as shown in other East African countries [An. gambiae s.l and An. funestus is difficult, and studies have shown great variability depending on the sampling technique used, and whether interventions such as ITNs are present and acting as a deterrent [An. arabiensis is also an important consideration as they prefer to feed on these animals over humans and other livestock [et al [An. gambiae s.l and An. funestus across a diverse ecological range using a standard sampling technique.The study by Mbogo et al  collecteountries . In geneeterrent ,28,47-49ivestock ,50,51. Ak [et al  is one oet al [An. gambiae and An. funestus complexes in one region of Kenya. It exemplifies what can be done with existing entomological data contained in the literature and elsewhere, and how modern satellite and GIS technologies in public health research may be exploited, especially for climate sensitive diseases in developing countries, such as malaria [Malaria transmission is complex, and more knowledge on the relationship between the environment, mosquito vectors, human disease and demography in sub-Saharan Africa will help implement appropriate control measures in a rapidly changing landscape. This is particularly important in areas already reporting changes in transmission intensity ,53, and et al  aimed to malaria ,55.The authors declare that they have no competing interests.LKH identified data sources, designed the study, carried out the data analysis and wrote the first draft of the manuscript. JH participated in the interpretation of the results and editing of the manuscript. EM conceived the idea for the study, and contributed to the writing and editing of the manuscript. All authors have read and approved the final manuscript.