Dengue fever is common in tropical and subtropical countries of the world and its present impact is 30 times more than what it was 50 yr ago. The aim of the study was to analyze the research output/publications on dengue across the world as well as in India, using the Web of Science database. Literature search was done using the keyword ‘dengue’ in the title field to find global and Indian publications on dengue from 1 June 1997 to 20 July 2018. Bibliometric analysis was done on the basis of the magnitude of research output; year, type and language of publications; citations and h-index, impact factor, keywords, most productive institutes etc. Descriptive analysis was performed using MS Office, Word Cloud Generator and SPSS software. The analysis of the records obtained indicated that globally 11,742 articles were published on dengue during the period of 21 yr of which 1114 were Indian. The country with highest number of publications was United States (3449; 29.37%) while India ranked third (1114; 9.48%). Majority of the published articles where in the American Journal of Tropical Medicine and Hygiene (1366; 11.63%) and the leading journal in India was Indian Journal of Medical Research (56; 5.02%) followed by the Journal of Vector Borne Diseases (42; 3.77%). The most productive Institute at global level was ‘Mahidol University, Thailand’ while in the Indian context it was ‘All India Institute of Medical Sciences, New Delhi’. The elevated metrics of citation, impact factor, and h-index are based on the magnitude of scientific research outputs. Increasing trend in the number of publications over the years both globally as well as in India indicates overall growth in dengue research. Even though it is a good sign of productivity it is still significantly related to the magnitude of occurrence of dengue cases across the globe. Further, it is a good indication that the scientific community is dynamic and responding well in bringing out the regional scenarios to the public for health promotion.

Malaria, a parasitic infectious disease causes approximately >1 million deaths annually worldwide. Treatment with effective antimalarials is one of the major strategies to combat malaria-related mortalities. However, there is a continuous threat of counterfeit antimalarials in the community. Counterfeit antimalarial drugs not only result in an economic loss but also decrease the efficacy of treatment resulting in the loss of faith in the health system and increases the the chances of drug resistance in the parasites. Counterfeit drugs hamper the intellectual property-based innovation paradigms as well. Awareness about these counterfeit drugs not only helps in avoiding drug resistance but may also enhance the drug therapeutic value. This review discusses the prevalence of counterfeit drugs in different geographic areas across the globe, the methods deployed for its detection and possible anticounterfeiting strategies. Literature search was conducted through PubMed, Google and International Pharmaceutical Abstracts using the terms ‘counterfeit antimalarials’, ‘substandard’, ‘falsified’, and ‘drug resistance’. Free searches in other search engines included the terms ‘antimalarial counterfeit drugs’ and ‘drug resistance’. Analysis of the literature survey indicated that majority of such studies were conducted in Southeast Asia and Africa region. The prevalence of substandard antimalarials was reported as high as 88.4% in Africa region and 53 % in Southeast Asia region. There is a need to follow a multifaceted approach to prevent the entry of falsified drugs with pre- and post-marketing surveillance. The samples need to be examined by regulatory bodies and strict legislation should be envisaged in order to maintain the quality of medicines.

Background & objectives: An outbreak of dengue-like illness was reported from Wadi area within the Nagpur Municipal Corporation during September–October 2017 with five deaths. Major symptoms reported were high fever (103–106 ^oF), acute joint pains, myalgia, drowsiness, breathlessness, etc. An investigation was conducted to confirm the etiological agent, its characterization and the vectors involved in the outbreak. Methods: Serological analysis was conducted to detect dengue (DEN)/chikungunya IgM antibodies in 158 sera samples. Nested-PCR was carried out to serotype eight ELISA positive samples. Adult and larval mosquito collections were conducted in the affected areas to determine species composition and mosquito density. Results: Dengue IgM antibodies were detected in 44 sera samples. Molecular typing revealed involvement of DEN-2 and DEN-3 serotypes. Dengue hemorrhagic fever symptoms were observed in two patients. Aedes aegypti breeding was found rampant with Breteu index and house index ranging from 23 to 70 and 17 to 56, respectively. Major breeding habitats encountered were, used tyres, cement tanks and refrigerator trays. Interpretation & conclusion: Clinical symptoms, detection of anti-DEN IgM antibodies in high number of samples and heavy breeding of Ae. aegypti confirmed it was a dengue outbreak.

Background & objectives: Interspecific competition occurs between members of two or more different species and can often have an influence on mosquito populations. Both Aedes aegypti and Anopheles stepehensi are container breeding mosquitoes and co-exist which may result in larval competition. In this study, interspecific competition between the above two species has been monitored under the laboratory conditions. Methods: Three sets of experiments were conducted with different stages of Ae. aegypti and An. stephensi larvae. First two experiments were set up with I/II instar and III/IV instar larvae of Ae. aegypti and An. stephensi respectively in the ratios of 20:20, 20:40 and 40:20 in plastic bowls. For third set of experiment 20 IV instar larvae of Ae. aegypti were put with equal number of I instar larvae of An. stephensi. Results: In the presence of food, 12.5–15 % mortality was recorded in I/II stage larvae of Ae. aegypti while in An. stephensi mortality ranged from 21–55%. Pupation commenced from Day 6 onwards in Ae. aegypti while in An. stephensi it commenced from Day 11 onwards. In the absence of food, there was no pupation in both the species but Ae. aegypti survived up to longer duration (7.5–18.5 days with 50% mortality) in comparison to An. stephensi (2–7 days with 50% mortality). When younger stages of An. stephensi (I/II) were put together with older stages of Ae. aegypti (III/IV) in the presence of food, pupation was completed in 85% Ae. aegypti population while there was 100% mortality in An. stephensi population. Interpretation & conclusion: The better survival and development of Ae. aegypti than An. stephensi under the same conditions exhibits interspecies competition showing competitive advantage of Ae. aegypti over An. stephensi. Further research is required to have a thorough understanding of the interaction between these two container inhabiting mosquito species in the nature.

Background & objectives: Prior to their elimination in 1974 and 2004, respectively, Plasmodium falciparum and P. vivax were the main native malaria parasites involved in disease transmission in Morocco. Imported cases of human malaria are still reported from the country. Anopheles labranchiae in northern Morocco and An. sergentii in the southern regions are the main malaria vectors. The bionomics and insecticide susceptibility of An. sergentii are poorly understood and need to be further studied to enhance the epidemiological surveillance of this important malaria vector. Methods: The adults and larvae of Anopheles sergentii were collected during the mosquito breeding season in 2015 and 2016 and environmental characteristics of their breeding sites were recorded. Blood meals were analyzed using PCR-RFLP. Alongside, the WHO routine susceptibility tests with DDT (4%) and malathion (5%) were conducted and An. sergentii specimens were screened for knockdown resistance (kdr) and acetyl cholinesterase encoding (ace-1) gene mutations. Results: Anopheles sergentii was observed during the summer and autumn seasons, feeding mainly on sheep, cows and also on humans. The WHO bioassays revealed complete susceptibility to DDT and malathion. Analysis of the sequences of the voltage-gated sodium channel gene revealed the absence of the kdr “Leu-Phe” mutation and PCR-RFLP revealed the absence of the G119S mutation in the ace-1. Interpretation & conclusion: With the increasing number of imported cases of human malaria in Morocco, the indiscriminate feeding behavior of this species may pose an infectious medical threat. Fortunately, the absence of insecticide resistance can ensure, for now, the efficiency of insecticides, as a part of the vector control program in controlling An. sergentii in Morocco.

Background & objectives: In India, kala-azar surveillance is weak and no public-private partnership exists for disease containment. Estimate of disease burden is not reliably available and still cases are going to private providers for the treatment. The present study aimed to assess the magnitude of kala-azar cases actually detected and managed at private set-up and unreported to existing health management information system. Methods: Institution based cross-sectional prospective pilot study was conducted. List of facilities was created with the help of key informants. The information about incidence of kala-azar cases were captured on monthly basis from July 2010 to June 2011. Rapid diagnostic strip test (rk-39) or bone marrow/splenic puncture were applied as laboratory methods for the diagnosis of kala-azar. Descriptive statistics as well as chi-square test for comparison between proportions was conducted. Results: Overall availability of private practitioners (PPs) was 4.59/1,00,000 population and maximum PPs (46; 93.9%) were from qualified category. The median years of medical practice was 25 yr (inter quartile-range [18, 28]). Interestingly, only a small proportion (240; 19%) of cases was managed by PPs. Amongst the PPs, only low proportion (32; 18.2%) managed >2 cases per month. The mean number of kala-azar suspects and cases identified varied significantly between different PPs’ professions with p <0.048 and p <0.032, respectively. A highly significant difference (p <0.0001) was observed for kala-azar case load between qualified and unqualified practitioners. A small proportion (38; 15.8%) of kala-azar cases was not present in the public health system record. Interpretation & conclusion: Still sizeable proportions of cases are going to PPs and unrecorded into government surveillance system. A mechanism need to be devised to involve at least qualified PPs in order to reduce treatment delay and increase case detection in the region.

Background & objectives: Generally filarial antigens have been found to be cross-reactive in nature. Identification of genus and species-specific antigens has not been successful so far. Due to lack of human adult filarial parasite, researchers have been using other adult worms like Setaria digitata, a cattle parasite or Brugia malayi, a rodent model for their research work. In this situation, specificity of the prepared antigen (S. digitata or B. malayi) to detect the antibodies to Wuchereria bancrofti is questionable. Methods: In the present investigation, we have tested a panel of human sera (collected from the areas, endemic for bancroftian filariasis) to correlate the immune reactivity against somatic antigens of adult stages and microfilarial stages of S. digitata and B. malayi. Further, using intact microfilariae (mf) from the above two parasites along with W. bancrofti, we have analyzed the antibody response to the sheath antigens. A panel of infected human and cattle sera was tested by immunoperoxidase assay using intact mf of three different parasites, viz. W. bancrofti, B. malayi, and S. digitata. Results: A very significant positive correlation in filarial Igs (polyvalent), IgG, IgM, IgE and IgG4 levels were found between the two adult somatic antigens of B. malayi and S. digitata when tested against human filarial sera. However, such a correlation was not found when mf antigens of B. malayi and S. digitata were tested against a panel of W. bancrofti sera indicating that antigens present in mf could be far less cross-reactive in comparison to those in adult stage parasites. Interpretation & conclusion: The results indicated the differential cross-reactivity of antisheath antibodies to the mf sheath of three different filarial parasites. Soluble antigens of S. digitata could inhibit antisheath antibody reactivity to only S. digitata mf sheath and not to mf sheath of W. bancrofti further confirming the specificity of sheath antigen.

Background & objectives: Leishmaniasis is a major global health problem with no safe and effective therapeutic drugs. This study evaluated the cytotoxic and apoptotic effects of crude extract and fractions of Gossypium hirsutum bulb on Leishmania major stages using advanced experimental models. Methods: Bulbs of G. hirsutum were collected from the Kerman province of Iran. The bulb was extracted using Soxhlet apparatus and different fractions were obtained by column chromatography (CC). Different concentrations of the extract and the fractions were evaluated against L. major and compared with Glucantime®. The cytotoxicity and apoptotic values were analysed by flow cytometry. The fractions obtained in CC were monitored by thin layer chromatography, and fractions with similar chromatographic patterns were mixed. Results: The extract and two fractions, F4 and F5 inhibited the proliferation of L. major promastigotes and amastigotes in a dose-dependent manner at 72 h post-treatment. No significant cytotoxic effects were observed for extract and fractions, as the selectivity index was over 1000, far beyond >10. The mean apoptotic values for L. major were superior to those of Glucantime^®. Interpretation & conclusion: Both the crude extract and fractions (F4 and F5) had significant antileishmanial effects on L. major stages, and were were superior relative to Glucantime^®. No cytotoxic effects were associated with the extract or fractions and they showed excellent apoptotic index, a possible mechanism behind inducing parasite death. Further investigations are essential to study the effect of G. hirsutum bulb fractions in animal model and clinical settings for planning strategies for the prevention and control of leishmaniasis.

Background & objectives: Understanding of malaria vector distribution and influence of climatic environments is essential for devising control strategies. The aim of the study was to study the bionomics of prevalent malaria vectors in three different settings for development of evidence-based sustainable malaria control strategy with special reference to vector control. Methods: Three villages with different eco-epidemiological settings like riverine-low malarious, riverine-high malarious and non-riverine high malarious villages were selected after baseline studies. Entomological aspects such as man hour density, per structure density, mosquito landing collections, sibling species identification, insecticide susceptibility status, parity rate, etc. were studied in these three villages following standard methods and techniques. The effect of these variables was analysed statistically. Results: Mosquito collections revealed the presence of three malaria vectors in the study villages, namely Anopheles culicifacies s.l., An. fluviatilis s.l. and An. stephensi (Diptera: Culicidae) with varying proportions and seasonal abundance. Densities of the principal malaria vector, An. culicifacies varied seasonally. Anopheles culicifacies was found resistant to DDT (4%), malathion (5%), lambda-cyhalothrin (0.05%) and alpha-cypermethrin (0.1%). Peak density of An. culicifacies was found during post-monsoon months starting from August-September to December in the high malarious villages. Interpretation & conclusion: The main vector control interventions should be planned in the post-monsoon months in these villages and suitable insecticide resistance management strategy should be followed as An. culicifacies was found resistant to DDT, malathion, alpha-cypermethrin and lambda-cyhalothrin in the study area.

Background & objectives: Bioreactors are practical tools that are used for economical, time-conserving and large-scale production of biomass from cell cultivation. They provide optimal environmental conditions such as pH and temperature required for obtaining maximum amounts of biomass. However, there is no evidence in the literature on the large-scale cultivation of Leishmania infantum parasites in the bioreactor. Therefore, the present study was undertaken to develop a new approach for obtaining L. infantum biomass by using pH and temperature controllable stirred bioreactor and to compare parasitic growth kinetics with classical method within erlenmeyers. Methods: In order to obtain parasite biomass, a newly developed pH and temperature controlled stirred bioreactor was used and its efficacy was compared with a graduated classical scale-up method. Growth kinetics of parasites within erlenmeyers and bioreactors were determined by evaluating promastigote numbers using haemocytometer. The graduated scale enlargement of culture was followed by T25 flask, T75 flask, and 1 L erlenmeyer, respectively. Results: Obtained results showed a 10-fold increase in the number of promastigotes within the conventional culture performed in 700 ml medium, while parasite numbers increased approximately 15 times due to initial inoculation amounts in the bioreactor culture performed in the 3.5 l medium. Thus, there was 7.5 times more biomass collection in bioreactor compared to classical method. Interpretation & conclusion: It is postulated that constant culture pH and temperature in the bioreactor extends cultivation time. This could lead to significant increase in parasite numbers. Hence, pH and temperature controllable bioreactors provided acquisition of sufficient amounts of biomass in contrast to classical methods. Therefore, this type of bioreactors may substitute classical culture methods in the production of antigenic molecules for vaccine development.

Background & objectives: Insufficient treatment of cutaneous leishmaniasis (CL) by conventional drugs is a major barrier in control strategies. This study was aimed to evaluate Glucantime efficacy and the susceptibility of Glucantime unresponsive and responsive CL isolates in the field and laboratory. Methods: Chi-square test (x^[2]) was used to determine the significance of difference between proportions in Glucantime-treated patients. The inhibitory activity of various concentrations of Glucantime against Leishmenia tropica stages was evaluated by a colorimetric cell viability MTT and macrophage assays. Mixed model, t-test and ANOVA were performed to determine the significance of difference between various concentrations of Glucantime unresponsive or responsive isolates and untreated control group and p <0.05 was defined as significant level. Altogether, 89.8% of the patients were cured by Glucantime, whilst 10.2% remained non-cured. Results: The overall Glucantime efficacy in different age groups and genders was similar. The IC₅₀ values of promastigotes and amastigotes for Glucanime unresponsive isolates were 2.1 and 2.6 times higher than the equivalent rates obtained for responsive cases, respectively. The overall mean number of amastigotes within macrophages in unresponsive isolates was significantly higher (32.68 ± 1.24) than that in responsive ones (18.68 ± 1.52, p <0.001). Glucantime unresponsive and responsive field isolates of anthroponotic CL (ACL) caused by L. tropica strongly correlated to in vitro assays. Interpretation & conclusion: Monitoring of Glucantime unresponsiveness by the health surveillance system is extremely important, where anthroponotic transmission occurs in humans. Hence, physicians should be aware of such clinical unresponsive presentations with ACL for antimonial therapeutic failure to improve management of disease in endemic regions.

Background & objectives: Majority of the studies on severe malaria in India have concentrated on falciparum and have been done in northern part. The objective of the study was to compare the clinical spectrum and laboratory profile among severe Plasmodium vivax, P. falciparum and mixed malaria patients admitted at a tertiary care center in southern India. Methods: This prospective, observational study was done in adult patients with severe malaria hospitalized in a tertiary care centre in southern India. Malaria was diagnosed by either quantitative buffy coat test or peripheral blood smear. In the cases of P. vivax malaria, an antigen detection test was done to rule out coexistent falciparum infection. Severe malaria was defined as per the WHO guidelines. The malaria severity score (MSS) was calculated for all patients based on the clinical features and laboratory parameters. Results: A total of 204 cases of severe malaria were studied. Among them, 105 (51.5%) had vivax infection, 30 (14.7%) had falciparum and 69 (33.8%) patients had mixed malaria. The mean age of the study population was 39.8±15.7 yr. The majority were males (71.6%). Hypotension and prostration were the most common complications noted in the patients, irrespective of species. The maximum mean MSS was found to be highest in falciparum malaria, followed by mixed malaria and vivax. In vivax malaria, majority of patients (71.4%) had one or two complications and only 28.57% of patients had three more complications, whereas in falciparum malaria, the majority (53.33%) had three or more complications. Around 44.93% of mixed infection malaria patients had three or more complications. The number of patients with multi-organ dysfunction (>2 complications) was significantly more in patients with falciparum infections compared to the remaining patients. Interpretation & conclusion: Severe malaria in south India is predominantly due to vivax. Hypotension and prostration were the most common complication of severe malaria irrespective of the plasmodium species. The entire spectrum of severe malaria complications described for falciparum are seen in severe vivax malaria.

Background & objectives: Scrub typhus is an under-reported rickettsial illness caused by Orientia tsutsugamushi which is transmitted by trombiculid mites. Serious complications are not uncommon and multiorgan dysfunction may develop leading to death. Paucity of data on the clinical spectrum and determinants of aftermath may be contributing to higher mortality in the region. A prospective study was done to describe the spectrum of organ dysfunction in serologically confirmed cases of scrub typhus and document predictors of adverse outcomes. Methods: This prospective study was carried out in patients diagnosed to have scrub typhus by IgM ELISA. The clinical features, investigations and complications among survivors were statistically compared to those in the deceased. Fisher’s exact test, t-test and logistic regression have been applied where appropriate. Results: The study population comprised of 123 patients. Majority of patients (62%) had one or more organ dysfunction. Ten patients (8.1%) did not survive. Complications documented were acute kidney injury (AKI) in 35%, hepatitis in 29.2%, acute respiratory distress syndrome (ARDS) in 26%, shock in 13%, meningitis in 5.7%, disseminated intravascular coagulation (DIC) in 2.6%, pancreatitis in 2.6% and myocarditis in 1.6%. Certain clinical features, biochemical parameters and complications had statistically significant correlation with the outcome. The mean SOFA score was considerably higher in those who did not survive. Interpretation &conclusion: Patients developing hepatic dysfunction, acute kidney injury and respiratory distress should be identified early and intensively monitored. The SOFA score can be utilized to assess the severity at admission and rapidly triage the sicker patients.

Background & objectives: Rift Valley fever virus (RVFV) is a vector-borne pathogen that causes serious outbreaks among livestock, and severe symptoms and mortality in humans. The virus is known to be widespread throughout African countries and Arabian peninsula. The aim of the present study was to investigate the seroprevalence of RVFV infection among human populations of Mersin province, Turkey. Methods: A region-wide serological survey was conducted on humans residing in rural and urban areas of Mersin province located in the subtropical mediterranean region of Turkey from July 2011- January 2014. Plasma samples were tested for the presence of anti-RVFV antibodies using commercially available indirect immunofluorescence assay. Results: The overall past infections were detected in 48 (4.9%) of the 977 human blood samples. The RVF virus- specific IgG positivity was detected in 33 (4.9%) of the 677 blood samples obtained from the urban area and in 15 (5%) of the 300 samples obtained from the rural area. There was no statistically significant difference in the distribution of RVFV IgG positivity rates between urban and rural areas (p = 0.933); though difference was significant between the rural areas (p = 0.029). Interpretation & conclusion: The study confirmed for the first time, the presence of the RVFV antibody in the urban and rural areas of mediterranean province of Mersin in Turkey, suggesting wide circulation of RVFV in the human population.

A dengue outbreak struck Baranagar municipality area of North 24 Parganas district, West Bengal in July 2016. This study presents the epidemiological and virology findings of this outbreak. The outbreak started from 17 July and continued until 17^th December. Total 1660 dengue cases (overall attack rate: 7/1000) and two deaths (case fatality rate: 1/1000) were reported. All age groups were affected. Out of the 213 blood samples collected from probable dengue cases and tested at Virus Unit, ICMR-National Institute of Cholera & Enteric Diseases (NICED), Kolkata, 163 (76%) tested seropositive through NS1 / IgM ELISA confirming dengue infection. DENV 1 was the predominant (53%, 44/83) serotype followed by DENV 2 (23%, 19/83), DENV 4 (15.6%, 13/83), DENV 3 (8.4%,7/ 83). Coexistence of dengue and chikungunya virus infection was detected. Major presentation of the confirmed dengue cases was fever with headache (95%), followed by arthralgia, myalgia, retro-orbital pain, abdominal pain, diarrhoea, and rash. Abundance of vector breeding spaces was observed in the area. Accordingly the health authorities were suggested to conduct frequent mass awareness campaigns involving community to reduce breeding sources during pre-monsoon and monsoon months. Further, adult mosquito control measures were also suggested throughout the year and inter-departmental coordination was recommended for prevention of such outbreaks in future.