Leishmaniasis is a parasitic disease of tropics, subtropics, and southern Europe which is classified as a neglected tropical disease. There is a paucity of knowledge for this disease especially in some parts of the world where leishmaniasis is endemic such as Cyprus island in the Mediterranean. This narrative review revealed cutaneous leishmaniasis (CL) and visceral leishmaniasis (VL) as the dominant type of human leishmaniasis in the island. Host and disease type identification studies based on cats, dogs and rats showed Leishmania infantum parasite to be the leading species among others with a seroprevalence rate ranging from 3.55% to 14.9% in selected hosts with an indication of dogs as the main animal reservoir. Lack of focus on potential wild animal hosts creates an important evidence gap to overcome for the purpose of controlling leishmaniasis. We aim to present asynoptical review of leishmaniasis to critically analyse currently available data in literature about this disease in Cyprus.

Background and objectives: The emergence of multi-drug resistant (MDR) strains of Plasmodium falciparum highlights the need to develop novel antimalarial drugs. Present study explores the in vivo antiplasmodial activity of ethanol leaf extract of Thalictrum foliolosum (ELETF) against lethal murine malaria. Methods: The acute toxicity of the extract was assessed by Limit test of Lorke. The suppressive activity of the extract was evaluated by Peter’s 4 day test. In vivo preventive and curative activity of ELETF was assessed by Peter’s method and Ryley and Peter’s method respectively. Biochemical assays were carried out using standard methods. Results: ELETF (1000 mg/kg) exhibited considerable in vivo schizontocidal activity with 67.11% chemosuppression on Day 5. The ED₅₀ of the extract was 579.56 mg/kg. ELETF also showed significant repository activity with 87.70% chemosuppression at 750 mg/kg, which was greater than pyrimethamine (78.78%). ELETF exhibited dose dependent chemosuppression in the curative test with maximum 70.06% chemosuppression (750 mg/kg). Maximum Mean Survival Time (MST) was 19.2±4.60 and 22.66±4.41 days respectively in the suppressive and curative test, which was extremely statistically significant (p<0.0005) in comparison to untreated control which died by Day 9 post inoculation. Biochemical analysis revealed the safety of ELETF to the hepatic and renal functions of the rodent host. Interpretation & conclusion: The study reports the antiplasmodial potential of Thalictrum foliolosum (ELETF) against Plasmodium berghei infection. The extract can be developed as a phytomedicine against malaria. Alternatively, the active components can be isolated as new lead compounds against the disease.

Background & objectives: In India, the burden of Plasmodium vivax malaria has been projected to be highest in some areas. This study investigated the efficacy and safety of fixed dose combination (FDC) of arterolane maleate (AM) 37.5 mg and piperaquine phosphate 187.5 mg (PQP) dispersible tablets and (not with) chloroquine in the treatment of uncomplicated vivax malaria in pediatric patients. Methods: This multicentric, open-label trial was carried out at 12 sites in India. A total of 164 patients aged 6 months to 12 years with P. vivax malaria were randomized in a ratio of 2:1 to AM-PQP (111 patients) or chloroquine (53 patients) arms. The duration of follow up was 42 days. Results: At 72 hours, the proportion of a parasitaemic and afebrile patients was 100% in both treatment arms in per protocol (PP) population, and 98.2% and 100% [95% CI: -1.8 (-6.33 to 5.08)] in AM-PQP and chloroquine arms, respectively, in intent to treat (ITT) population. The efficacy and safety of AM-PQP was found to be comparable to chloroquine in the treatment of uncomplicated P. vivax malaria in pediatric patients. Overall, the cure rate at Day 28 and 42 was >95% for both AM-PQP or CQ. The commonly reported clinical adverse event was vomiting. No patient was discontinued for any QTc abnormality. Interpretation & conclusion: The efficacy and safety of FDC of arterolane maleate and piperaquine phosphate was found to be comparable to chloroquine for treatment of uncomplicated P. vivax malaria in pediatric patients.

Background & objectives: The Department of Health Research and the Indian Council of Medical Research, Government of India, have established Virus Research and Diagnostic Laboratory Network (VRDLN) to strengthen the laboratory capacity in the country for providing timely diagnosis of disease outbreaks. Fifty-one VRDLs were functional as on December 2017 and had reported about dengue fever across Indian states. The objectives of the study were to detect space time clusters and purely temporal clusters of dengue using Kulldorff's SaTScan statistics using patient level information; and to identify regions at greater risk of developing the disease using Kriging technique aggregating at district level. Methods: A total of 211,432 patients from 51 VRDLs were investigated for IgM antibodies or NS1 antigen against dengue virus during the period from 1 January 2014 to 31 December 2017 and among them 60,096 (28.4%) were found to be positive. Kulldorff's space time analysis was used to identify significant clusters over space and time. Kriging technique was used to interpolate dengue data for areas not physically sampled using the relationship in the spatial arrangement of the data set. Maps obtained using both the methods were overlaid to identify the regions at greater risk of developing the disease. Results: Kulldorff Space time Scan Statistics using the Bernoulli model with monthly precision revealed eight statistically significant clusters (P <0.001) for the time period, 1 January 2014 to 31 December 2017. Eight significant clusters identified were districts of Nagpur, Jhunjhunu, Gadag, Dakshin Kannada, Kancheepuram, Sivaganga, Ernakulam and Malda. The purely temporal clusters occurred during the last quarter of 2015 and 2016. The Kriging technique identified north eastern part of the country (Arunachal Pradesh, Nagaland and Manipur) and Gujarat. Interpretation & conclusion: Dengue fever has spread in all directions in the country. Hence, it is need of the hour to perform an in-depth investigation.

Background & objectives: Babesiosis is a tick transmitted disease, infecting a wide variety of wild and domestic animals, as well as humans. This study was designed to investigate molecular diagnosis and clinic-hemato-biochemical and oxidant/antioxidant status in dogs of Mizoram, India. Methods: A total 1200 dogs screened for babesiosis during 2017–18 and 53 dogs suspected for babesiosis by clinical signs and were confirmed by molecular diagnosis. Clinical signs were recorded; also blood samples were taken to investigate hematologic changes, serum biochemical variations and oxidative stress biomarkers. Results: The overall incidence of babesiosis in dogs of Aizawl, Mizoram, India during the study period recorded was 1.25% (15/1200) and 28.3% cases confirmed from 53 suspected dogs (15/53). The most commonly observed clinical signs were fever, emaciation, depression and icterus and lymphadenopathy. Significant reduction in PCV, HB, RBCs, MCHC, total protein, and albumin along with significant increase in MCV, WBCs, monocytes and BUN were the most consistent hemato-biochemical changes. Oxidant/antioxidant assessment showed significant reduction in superoxide dismutase, catalase and total anti-oxidant (TAC) along with significant increase in lipid peroxidase (LPO) activities. Interpretation & conclusion: The findings of this study demonstrated that the main causative agent of babesiosis in dogs in Mizoram Province is Babesia gibsoni which caused significant alteration of hemato-biochemical and oxidant-antioxidant status in dogs.

Background & objectives: Aedes vittatus (Bigot), an anthropophilic mosquito, plays an important role in the maintenance and transmission of yellow fever (YF), dengue (DEN), chikungunya (CHIKV) and Zika (ZIK) viruses in Africa. In India, though natural isolation of none of these viruses was reported from the mosquito, experimental studies have shown vector competence to DEN and CHIK viruses. Despite wide prevalence in India, their potential in transmitting viruses of public health importance viz., Japanese encephalitis (JEV), West Nile (WNV), Chandipura (CHPV), Chittoor (CHITV) etc., has never been investigated. The objective of the present study is to determine the vector potential of the mosquito to these viruses. Methods: Mosquitoes were infected by intra-thoracic inoculation as well as by oral feeding, and growth kinetics was determined. Virus dissemination to organs was investigated by determining virus in the harvested organs on specified days’ post infection (PI). Vector competence was determined by detecting the virus in saliva. Results: Intra thoracic inoculation has shown vector competence of the mosquito to JEV, WNV, CHIV and CHPV. However, using the oral route of infection, replication was observed with only WNV, JEV and CHITV. High degree of WNV replication (6.7log TCID₅₀/ml) with rapid dissemination to wings, legs and salivary glands was seen from 5^th day PI onwards. WNV was detected in saliva with a titer of 0.7log₁₀ TCID₅₀/ml on 5^th day PI. JEV and CHITV replicated in the mosquito yielding 3log and 4log₁₀ TCID₅₀/ml on 5^th and 10^th day PI respectively, but virus was not detected in saliva till 15^th day PI. Interpretation & conclusion: From the results it is difficult to indict the mosquito as a vector of the viruses studied. However, presence of WNV in saliva of the mosquito shows its potential as a bridge vector and poses a concern especially when virulent WNV strains are circulating in the country.

Background & objectives: Indoor residual spraying (IRS) is part of a key strategy for elimination of visceral leishmaniasis (kala-azar). IRS for kala-azar elimination in India uses 125 g 5% alpha-Cypermethrin wettable powder which is mixed with 7.5 litres of water and sprayed on walls using hand compression sprayers. Insecticide quantity is measured volumetrically through a container. Methods: A cross-sectional study design with cluster random sampling was adopted to select 272 IRS squads of 46 blocks across 12 districts in Bihar, India. The quantity of insecticide measured by the container used by each IRS squad was recorded. Results: Mean weight of insecticide measured was found to be 147 g (SD 33.9). One-thirds of squads were measuring less than the nationally recommended quantity of 125 g. Two-fifths of squads were overdosing with use of more than 150 g insecticide powder. Shoving the containers into a heap of insecticide powder resulted in heavier and less consistent measurements as compared to filling the containers from the top. Different types of measuring containers and different manufacturers of insecticide were shown to significantly account for the variation in the quantity of insecticide being measured. Interpretation and conclusion: Standardization of insecticide measurement by IRS squads is needed, both to prevent under-dosing and overdosing of insecticide residue on walls. Standard operating procedures for calibrating and using uniform measuring containers should be implemented. Further, use of measuring containers may be replaced altogether with manufacturer-packaged amounts of insecticide formulation which could be directly used to prepare one-tank load of insecticide suspension.

Background & objectives: The global incidence of dengue has grown dramatically in recent decades and Assam, India has witnessed several outbreaks of dengue since 2015. Although during post-monsoon months (September to December), most cases of dengue in Assam are recorded but incidence of dengue in Assam has been slowly changing from being endemic to being hyper endemic. Therefore, this study was carried out to determine the serotypes and genotypes of dengue virus prevalent in Assam during the period of 2016–2017. Methods: This is a prospective study conducted for a period of two years from 2016 to 2017. Department of Microbiology, Gauhati Medical College and Hospital (GMCH) had received a total of ~12000 and ~9000 sera sample during 2016 and 2017 respectively for confirmation of clinically suspected dengue cases. For confirmation, dengue NS1 antigen and IgM antibody ELISA tests were performed. Multiplex RT-PCR was performed for serotyping of dengue viruses and representative samples found positive in PCR were sequenced to determine the genotypes of circulating dengue virus serotypes. Results: In the year 2016, 6157 sera samples and in 2017, 3386 sera samples were found positive in ELISA test. A total of 157 dengue positive sera samples representing 17 districts of Assam were further tested by multiplex RT-PCR for serotyping of the virus. In PCR, out of 157, 107 samples (68.15%) were found positive for the presence of dengue virus genome. Out of 107, 74 samples (69.15%) were positive for dengue virus serotype-1 (DENV-1), 32 samples (29.90%) for dengue virus serotype-2 (DENV-2) and one sample (0.93%) positive for dengue virus serotype-3 (DENV-3). Out of 107 PCR positive samples, 25 samples were sequenced to identify their genotypes. Phylogenetic analysis of sequenced dengue viruses revealed that all the seven DENV-1 strains were genotype V, 17 DENV-2 strains were genotype IV (Cosmopolitan genotype) and one DENV-3 strain was genotype III. Interpretation & conclusion: These findings improve our knowledge of circulating dengue virus serotypes in Assam. Co-circulation of three serotypes of dengue virus highlights the need for establishment of active dengue surveillance. The genotypic data of our findings will be helpful for future dengue molecular epidemiology studies and to control the disease in the region.

Background & objectives: Culex tritaeniorhynchus is a widely distributed and medically important mosquito species (Diptera: Culicidae). The aim of this study was to determine characteristics of larval habitats and predict the distribution of Cx. Tritaeniorhynchus using maximum entropy (MaxEnt) model in Golestan Province (north of Iran). Methods: Overall, 53 villages and cities from the 14 counties in the Golestan Province were studied. Suitable habitats for sampling of mosquitoes were determined in the selected villages. Larval sampling was carried out 10 times using a standardized dipper (350 ml). Captured larvae in each dipper were counted and kept in special containers. Larvae were put in lactophenol and were then mounted on microscope slides prepared according to the de Faure’s formula. All villages where Cx. tritaeniorhynchus was isolated from were considered as the species presence points, and their coordinates were extracted from the GIS. The obtained data were entered into the model in Microsoft Excel 2003. Layers required for the study were obtained from the World Clim global climate database at a spatial resolution of 1 km². The MaxEnt software (version 3.3.3) was used to predict the most suitable ecological niches of Cx. tritaeniorhynchus. Results: Out of the 5168 specimens collected from 53 villages and cities, 1097 specimens (21%) were identified as Cx. tritaeniorhynchus. The central and western areas of the province are environmental suitability areas for distribution of this species compared to the other areas. Area under curve (AUC) value for Cx. tritaeniorhynchus was 0.888. Environmental variables with highest gain in the model were precipitation of coldest quarter (bio19), maximum temperature of warmest month (bio5) and slope. Interpretation & conclusion: Precipitation and temperature are important determinants of Cx. tritaeniorhynchus distribution in Golestan Province.

Background & objectives: For detection and molecular characterization of Babesia microti in laboratory mice from India. Methods: A total of 625 mice were screened by peripheral blood smear examination and subsequently was confirmed by PCR using a piroplasm conserved primer set (Piro A/B). Nested PCR was done using a species-specific primer targeting the gene encoding the small subunit ribosomal RNA (18S rRNA). The PCR products were cloned, purified and sequenced. A total of 12 isolates were obtained. The sequences were aligned and phylogenetic trees were prepared with other published Babesia spp. sequences. Results: B. microti was detected with a total infection rate of 8.80%. The higher rate of infection was observed by species specific PCR (8.80%) than examined by blood smear (7.20%). Sequence and phylogenetic analysis showed that Babesia species detected in mice were genetically identical to the genotypes of B. microti and can be easily distinguished from other genotypes of Babesia parasites by neighbour joining and maximum likelihood method. Intra-species analysis indicated that all the twelve isolates from six North-Eastern states of India have a close identity but inter-species showed genetic reservoir host for transmission of babesial infection to humans. Interpretation & conclusion: The detection of Babesia microti may suggest that laboratory mice may serve as potential reservoir host for human infection and possibility of innovative way of diagnosing and control of human babesiosis.

Hyperreactive malarial splenomegaly (HMS) is one of the important causes of massive splenomegaly in malaria endemic zones. It is thought to represent a dysfunctional immune response to recurrent malarial infection. It is usually reported due to physical symptoms of splenomegaly and hypersplenism and fever is classically absent. Concomitant malaria with HMS is a very rare finding in the Indian context. Here, we report a case of symptomatic falciparum malaria presented with fever, jaundice, massive splenomegaly and pancytopenia. Persistent massive splenomegaly led us to investigate thoroughly and finally diagnosed it as HMS with concomitant falciparum malaria. He received standard antimalarial treatment and 12 months of weekly chloroquine and completely recovered without any relapse or complications.

Background & objectives: Rickettsioses are zoonoses transmitted to humans by arthropods. They are due to strict intracellular bacteria belonging to the family Rickettsiaceae. Our purpose is to present the clinical and paraclinical characteristics of 14 new cases diagnosed in Al-Hoceima region, Morocco; indeed, the patients associated a diagnosis of Mediterranean Spotted Fever (MSF). Results: The average age of patients was 55 years. The patients were hospitalized for infectious syndrome, renal deficiency, pneumonia, and suspected meningitis. All cases had a general papular rash with palmo-plantar involvement, 12 out of 14 patients showed an escarotic spot, while neurological disorders were observed in 2 patients. Ophthalmic involvement was represented by retinal vasculitis in a single patient. Thrombocytopenia and cytolysis were constant in all patients. Renal deficiency was found in 3 cases and 2 cases had interstitial syndrome. The serology was positive in only one patient. All cases had been treated with doxycycline 200mg/day for 7 days with a good improvement of the clinical and biological symptoms. Interpretation & conclusion: The frequency of MSF in Morocco is not completely elucidated. A good knowledge of the clinical form allows an early diagnosis in order to institute an effective treatment.