Efficacy and safety of Once-Daily Vilanterol/Fluticasone furoate MDI in persistent asthma: Phase 3 OD-INHALE Study.

INTRODUCTION: Once-daily inhalers have been shown to improve adherence leading to lesser discontinuation compared to twice- or thrice-daily inhalers in management of asthma. Combination of Vilanterol and Fluticasone Furoate (VI/FF) is approved for management of asthma and COPD and is available as a dry powder inhaler. Pressurized-Metered Dose Inhalers (pMDIs) offer ease-of-use and therapy alternatives for patients with low inspiratory flow. This study assessed the efficacy and safety of a new once-daily pMDI containing VI/FF in individuals diagnosed with persistent asthma. METHODS: This phase 3, double-blind, randomized controlled study assessed the non-inferiority of VI/FF (12.5 mcg/50 mcg & 12.5 mcg/100 mcg; 2 puffs once-daily) over Formoterol Fumarate and Fluticasone Propionate (FOR/FP, 6 mcg/125 mcg & 6 mcg/250 mcg; 2 puffs twice-daily) in patients with persistent asthma. Primary outcome was change from baseline in trough FEV1 at the end of study (12 weeks). Adverse events and number of exacerbations were used to evaluate safety. RESULTS: A total of 330 patients were randomized into VI/FF (165) and FOR/FP (165). Trough FEV1 significantly improved in both the groups at week 12, with a mean difference (VI/FF minus FOR/FP) being 54.75 mL (95% CI, 8.42-101.08 mL, p = 0.02). The low dose VI/FF had similar efficacy to that of low dose FOR/FP and high dose VI/FF had similar efficacy to high dose FOR/FP. No serious adverse events were reported during the study. CONCLUSION: Once daily VI/FF pMDI was non-inferior to twice daily FOR/FP pMDI in patients with persistent asthma.

A review on pathogenicity of Aeromonas hydrophila and their mitigation through medicinal herbs in aquaculture.

Aeromonas hydrophila is a freshwater, facultatively anaerobic, chemo-organoheterotrophic bacterium that distressed fishes with gastroenteritis, septicemia and causes a disease known as Motile Aeromonas Septicemia (MAS), which affects the aquatic environment. Haemolysin, aerolysin, cytosine, gelatinase, enterotoxin and antimicrobial peptides have been identified as virulence factors in A. hydrophila. Medicinal herbs/plants and their uses are the instant, easily available, cost-effective, efficient and eco-friendly approach for socio-economic, sustainable development of modern aquaculture practice. Phytotherapy either through a dip or by incorporation into the diets is an alternative approach to synthetic pharmaceuticals to diminish the pathogenicity of aquatic environmental pathogens. Due to the presence of remarkable phytoconstituents like flavonoids, alkaloids, pigments, terpenoids, steroids and essential oils, the medicinal plant exhibits anti-microbial, appetite-stimulating, anti-stress, growth-promoting and immunostimulatory activities. Aqua-industry preferred phytotherapy-based techniques/compounds to develop resistance against a variety of aquatic pathogens in culturable fishes because they are inexpensive and environment-friendly. As a result, this review elaborates on the diverse applications of phytotherapy as a promising tool for disease management in aquaculture and a major step toward organic aquaculture.

Impact assessment of Intensified Malaria Control Project in transitioning a high malaria-endemic district to a low-endemic district: an epidemiological aspect.

Intensified Malaria Control Project (IMCP) was implemented in 2005 to control malaria in all North-Eastern and Odisha states of India. The present study aimed to investigate the impact of IMCP in reducing the malaria burden in Udalguri district, Assam state of North-East India. Malaria epidemiological data were obtained for IMCP intervention (Udalguri) and nonintervention district (West Singhbhumi, Jharkhand state). IMCP activities include introducing bi-valent rapid diagnostic kits (RDTs), Artemether-Lumefantrine drug in North-East India, long-lasting insecticidal nets (LLINs) distribution, and creating awareness programs about malaria in an intensified mode. The data revealed a significant decline in annual parasite incidence (API) from 14.94 (2005) to 2.61 (2018), -37% (95%CI: -57%, -19%, p = 001) after using LLINs in 2009 and -64% (95%CI: -116%, -14%, p = 013) after the introduction of RDTs in district Udalguri. Whereas control district showed a -28% (95%CI: -63%, 6.3%, p = 0.051) decrease in API using LLINs and a 10% (95%CI: -7.6%, 28%, p = 0.122) increase after the introduction of RDTs. Plasmodium falciparum (Pf) and P. vivax (Pv) were the major malarial parasites in Udalguri. Pv-malaria was much higher (71%) than Pf-malaria (29%) during the study period. An increasing trend of Pf cases was observed in Udalguri. Udalguri and Khoirabari BPHCs showed an overall reduction of 94% (95%CI: -143%, -45%, p = 0.001) and 84% (95%CI: -126%, -39%, p = 0.003), respectively; however, only a 10% (95%CI: -65%, -41%, p = 0.360) reduction in API was observed in Orang BPHC. An overall decrease in malaria indicates the effective implementation of vector and disease control strategies in the Udalguri district.

Photocatalytic degradation of tetracycline antibiotic and organic dyes using biogenic synthesized CuO/Fe2O3 nanocomposite: pathways and mechanism insights.

Tetracycline (TC) is a frequently administered antibiotic in many countries, due to its low price and excellent potency. However, certain antibiotics can be hazardous to living creatures due to their accumulation by complexation with metal ions which can contribute to teratogenicity and carcinogenicity. In this investigation, copper oxide-ferric oxide nanocomposite (CuO/Fe2O3 nanocomposite) was synthesized employing Psidium guajava (P. guajava) leaf extract as a reductant as well as a capping agent in an environment friendly and economical green synthesis method. The as-synthesized CuO/Fe2O3 nanocomposite was comprehensively characterized using various sophisticated techniques and its efficiency as a photocatalyst for degradation of tetracycline (TC) antibiotic and toxic dyes, i.e., rhodamine B (RhB) and methylene blue (MB) were investigated. The CuO/Fe2O3 nanocomposite exhibited exceptional efficiency for degradation of TC antibiotic (88% removal in 80 min), RhB (96% removal in 40 min), and MB (93% elimination in 40 min) with apparent rate constant of 0.048, 0.068, and 0.032 min-1, respectively. In the degradation experiments, photocatalytic activity of CuO/Fe2O3 nanocomposite was studied by varying different factors such as time of contact, catalyst dose, and solution pH. The role of reactive species in antibiotics and dye degradation was validated by radical scavenging studies which indicated that.OH radical played a critical role in photocatalytic decomposition. Furthermore, liquid chromatography-mass spectrometry (LC-MS) investigations were employed to anticipate a plausible mechanism for TC degradation.

Relativistic Spin Magnetohydrodynamics.

Starting from the kinetic theory description of massive spin-1/2 particles in the presence of a magnetic field, equations for relativistic dissipative nonresistive magnetohydrodynamics are obtained in the small polarization limit. We use a relaxation-time approximation for the collision kernel in the relativistic Boltzmann equation and calculate nonequilibrium corrections to the phase-space distribution function of spin-polarizable particles. We demonstrate that our framework naturally leads to emergence of the well-known Einstein-de Haas and Barnett effects. We obtain multiple transport coefficients and show, for the first time, that the coupling between spin and magnetic field appear at gradient order in the hydrodynamic equation.

Moderate Rainfall and High Humidity During the Monsoon Season, Negligence in Using Malaria Protection Methods and High Proportion of Mild Symptomatic Patients Were the Driving Forces for Upsurge of Malaria Cases in 2018 Among Tea Tribe Populations in Endemic Dolonibasti Health Sub-center, Udalguri District, Assam State, North-East India.

Malaria elimination is a global priority, which India has also adopted as a target. Despite the malaria control efforts like long-lasting insecticidal nets distribution, rounds of indoor residual spray, the introduction of bi-valent rapid diagnostic tests and artemisinin combination therapy, malaria remained consistent in Dolonibasti sub-center of Orang block primary health center (BPHC) under the district Udalguri, Assam state followed by abrupt rise in cases in 2018. Therefore, we aimed to investigate the factors driving the malaria transmission in the outbreak area of Dolonibasti sub-center. Malaria epidemiological data (2008-2018) of Udalguri district and Orang BPHC was collected. The annual (2011-2018) and monthly (2013-2018) malaria and meteorological data of Dolonibasti sub-center was collected. An entomological survey, Knowledge, Attitude and Practices study among malaria cases (n = 120) from Dolonibasti was conducted. In 2018, 26.1 % (2136/ 8188) of the population of Dolonibasti were found to be malaria positive, of which 55% were adults (n = 1176). Majority of cases were from tea tribe populations (90%), either asymptomatic or with fever only, 67.5 % (81/120) had experienced malaria infection during past years. The outbreak was characterized by a strong increase in cases in June 2018, high proportion of slide falciparum rate of 26.1% (other years average, 15.8%) and high proportion of P. falciparum of 81.2 % (other years average, 84.3%). Anopheles minimus s.l. was the major vector with 28.6% positivity and high larval density in paddy fields/ drainage area. Annual relative humidity was associated with rise in malaria cases, annual parasite incidence (rs = 0.69, 90%CI; p = 0.06) and slide positivity rate (rs = 0.83, 95%CI; p = 0.01). Older people were less educated (rs = -0.66; p < 0.001), had lesser knowledge about malaria cause (rs = -0.42; χ2=21.80; p < 0.001) and prevention (rs = -0.18; p = 0.04). Malaria control practices were followed by those having knowledge about cause of malaria (rs = 0.36; χ2 = 13.50; p < 0.001) and prevention (rs = 0.40; χ2 = 17.71; p < 0.001). Altogether, 84.6% (44/52) of the respondents did not use protective measures. We described a sudden increase in malaria incidence in a rural, predominantly tea tribe population group with high illiteracy rate and ignorance on protective measures against malaria. More efforts that are concerted needed to educate the community about malaria control practices.

Study of Clinical, Hematological and Molecular Characteristics of Patients of Thalassemia and Hemoglobinopathies in Tertiary Care Hospital.

To study clinical, hematological and molecular characteristics of patients of thalassemia and hemoglobinopathies and to correlate the molecular characteristics with clinical and hematological presentations. Material: This observational cross sectional study included 100 patients of age >12 years of all genders with chronic haemolytic anemia and history of multiple blood transfusion. Blood and radiological investigations were done. Clinical, hematological and molecular characteristics were studied. Observation and Clinical: Pallor was present in all cases and icterus in 32% cases. Total 48% of the patients had hepatomegaly and 98% had splenomegaly. Among genotypes, 15% cases had α-thalassemia, 62% had ß thalassemia + 뫧 thalassemia, 7% had HbS hemoglobinopathy, and 16% had HbE hemoglobinopathy. Hematological: Hemoglobin showed significant association with molecular genotypes of thalassemia with lowest being present in ß-thalassemia + 뫧 thalassemia and HbE.MCV showed significant association with molecular genotypes, with HbE having the lowest MCV of 65.5 fl. LDH levels showed a significant association with molecular genotype with highest being in HbS hemoglobinopathy. Molecular Characteristics: Common mutations in compound α-thalassemia were 3.7, 4.2 and 20.5 deletion. As for ß-thalassemia and 뫧 thalassemia, 47 cases had heterozygous type and 15 cases had homozygous types. In ß-thalassemia, the homozygous type showed IVS1- 5(G→C),CD 41/42(→CTT) and IVSII-654(G→T) while heterozygous type showed CD16(→G), CD 41/42(→CTT), IVS1-5(G→C), and IVSII-654(G→T) . In 뫧 thalassemia, the heterozygous type showed 뫧 inversion mutation.In HbS hemoglobinopathy, heterozygous type showed Codon 6(A→T) and compound heterozygous type showed IVS1- 5(G→C) and Codon 6(A→T). In HbE hemoglobinopathy,the homozygous type showed CD26(G→A) and compound heterozygous type showed IVS1-5(G→C) and IVSII 654(G→T). Conclusion: The common thalassemia genotypes observed in our study were α-thalassemia (15%), ß thalassemia + 뫧 thalassemia, (62%) HbS hemoglobinopathy (7%), and HbE hemoglobinopathy (16%). The patients presented with pallor, icterus, hepatomegaly, and splenomegaly which were comparable among all molecular genotypes of thalassemia and hemoglobinopathies. α-thalassemia had compound α-thalassemia with common mutations being 3.7, 4.2 and 20.5 deletion. As for ß-thalassemia and 뫧 thalassemia, 47 cases had heterozygous type and 15 cases had homozygous types. In 뫧 thalassemia, the heterozygous type showed 뫧 inversion mutation in 5 cases. MCH, Retic count, ferritin stores, and peripheral blood smear were similar in all molecular genotypes. Hemoglobin, MCV and LDH showed a significant association with molecular genotypes. Microcytic hypochromic anaemia was commonest among all.The findings of the present study show that the genotypes of thalassemia are characterized by diversity as well as significant genetic heterogeneities.

Malaria Epidemiology Along the Indian Districts Bordering Bhutan and Implications for Malaria Elimination in the Region.

It is important for malaria-endemic countries to address malaria control across international borders, and in particular to prioritize appropriate rapid diagnosis, treatment and surveillance. Bhutan and India aim to achieve malaria elimination by 2023 and 2030 respectively. Malaria elimination along the Indo-Bhutan border is of common concern. We delineated malaria epidemiology along the border to provide a blueprint for focusing malaria control efforts in key foci within this region. Epidemiological data from 2015 to 2019 were analyzed, as the most drastic reductions in malaria burden across most parts of India were witnessed in this time frame. Several areas of concern include low surveillance in most border districts, favorable climatic conditions for perennial malaria transmission, and movement of potential parasite carriers because of the porous borders. India and Bhutan need to control the importation/exportation of malaria cases. We highlight the foci of concern for which implementing tailor-made malaria control strategies may benefit both countries.

Therapeutic efficacy of artemether-lumefantrine for the treatment of uncomplicated Plasmodium falciparum malaria in four malaria endemic states of India.

BACKGROUND: Malaria is a major public health problem in India and accounts for about 88% of malaria burden in South-East Asia. India alone accounted for 2% of total malaria cases globally. Anti-malarial drug resistance is one of the major problems for malaria control and elimination programme. Artemether-lumefantrine (AL) is the first-line treatment of uncomplicated Plasmodium falciparum in north eastern states of India since 2013 after confirming the resistance against sulfadoxine-pyrimethamine. In the present study, therapeutic efficacy of artemether-lumefantrine and k13 polymorphism was assessed in uncomplicated P. falciparum malaria. METHODS: This study was conducted at four community health centres located in Koraput district of Odisha, Bastar district of Chhattisgarh, Balaghat district of Madhya Pradesh and Gondia district of Maharashtra state. Patients with uncomplicated P. falciparum malaria were administered with fixed dose combination (6 doses) of artemether-lumefantrine for 3 days and clinical and parasitological response was recorded up to 28 days as per World Health Organization protocol. Nucleotide sequencing of msp1 and msp2 gene was performed to differentiate between recrudescence and reinfection. Amplification and sequencing of k13 propeller gene region covering codon 450-680 was also carried out to identify the polymorphism. RESULTS: A total 376 malaria patients who fulfilled the enrolment criteria as well as consented for the study were enrolled. Total 356 patients were followed up successfully up to 28 days. Overall, the adequate clinical and parasitological response was 98.9% and 99.4% with and without PCR correction respectively. No case of early treatment failure was observed. However, four cases (1.1%) of late parasitological failure were found from the Bastar district of Chhattisgarh. Genotyping of msp1 and msp2 confirmed 2 cases each of recrudescence and reinfection, respectively. Mutation analysis of k13 propeller gene showed one non-synonymous mutation Q613H in one isolate from Bastar. CONCLUSIONS: The study results showed that artemether-lumefantrine is highly effective in the treatment of uncomplicated P. falciparum malaria among all age groups. No functional mutation in k13 was found in the study area. The data from this study will be helpful in implementation of artemether-lumefantrine in case of treatment failure by artesunate plus sulfadoxine-pyrimethamine.

Evaluation of the acute toxicity of Thymus linearis ethanol extract and its effect on the hemato-biochemical and behavioural response of the Golden mahseer, Tor putitora (Hamilton, 1923).

The present investigation was conducted to estimate the acute toxicity of Thymus linearis plant extract, its effect on hemato-biochemical parameters and behavioural response in the golden mahseer (Tor putitora). The phytochemical composition present in T. linearis plant extrat were Alkaloids, Flavonoids, Phenols and Tannin. The fishes were subjected to eight different concentrations of T. linearis leaves extract (8.25, 8.50, 8.75, 9.00, 9.25, 9.50 and 9.75 mg/kg) and the control group without plant extract for 96-h LD50 study. The mortality was recorded every 24 h post-treatment. Minimum mortality was recorded in the 8.25 mg/kg, whereas 100% mortality was recorded in the 9.75 mg/kg T. linearis extract after 96-h periods. The LD50 was estimated by probit analysis, and the value of T. linearis at 96 h was found to be 8.71 mg/kg for golden mahseer. A non-lethal dose of 1/10th of 96-h LD50 value (0.87 mg/kg) was taken for the sublethal study. After 96 h, the red blood cell (RBC), white blood cell (WBC), haemoglobin (Hb), packed cell volume (PCV) and blood glucose were measured. RBC (×106/mm3), Hb (%) and PCV (%) significantly decreased at 8.25, 8.50, 8.75, 9.00 mg/kg, but WBC and blood glucose significantly increase at 8.25, 8.50, 8.75, 9.00 mg/kg of T. linearis plant extract. The observations on behaviour response of golden mahseer were also recorded. In the present study, the acute toxicity of wild ajwain was more significant than short-term toxicity. The mortality rate was very high during the study period of T. linearis exposure.

Clinical and molecular monitoring of Plasmodium falciparum resistance to antimalarial drug (artesunate+sulphadoxine-pyrimethamine) in two highly malarious district of Madhya Pradesh, Central India from 2012-2014.

The spread of P. falciparum resistant strain has led to a significant resurgence of malaria morbidity and mortality. The current cornerstone in malaria treatment in India is Artemisinin based Combination (Artesunate + Sulphadoxine-Pyrimethamine) Therapy (ACT) for treatment of uncomplicated P. falciparum malaria since 2010. In the present study we assessed the therapeutic efficacy of ACT and molecular monitoring of antimalarial resistance. Therapeutic efficacy was determined by in vivo method using 28 days follow-up. Molecular genotyping of dihydrofolate reductase (dhfr), dihydropteroate synthase (dhps) and kelch13 genes were analyzed. msp-1 and msp-2 genotyping were used to differentiate recrudescence. Therapeutic efficacy of ACT was determined in 237 patients over the three year period. Most of the patients showed adequate clinical and parasitological response (99.6%). Molecular study revealed that 72% parasites were of mutant genotype (27.2% single mutants, 43.5% double mutants and 1.3% triple mutants) for pfdhfr while pfdhps showed 78.2% wild type alleles and 21.8% mutants (18.1% single mutants and 3.7% double mutants). Analysis of total 135 samples revealed mutation in k13 gene along with non-synonymous single mutation at codon M579T (1.5%) and double mutations at codon M579T & N657H in 37%. ACT remains effective for the treatment of uncomplicated P. falciparum malaria in Madhya Pradesh, Central India. However, increasing mutation in pfdhfr (particularly triple mutations) and pfdhps may reduce susceptibility to partner drug SP and mutation in k13 propeller gene, highlighting the need for continuous monitoring of the efficacy of ACT.

Development and optimization of gastroretentive mucoadhesive microspheres of gabapentin by Box-Behnken design.

CONTEXT: Gabapentin follows saturation kinetics for absorption because of carrier-mediated transport and narrow absorption window in stomach. There is need to develop a gastroretentive formulation to maximize the absorption without crossing the saturation threshold for absorption. OBJECTIVE: The aim was to develop a gastroretentive formulation of gabapentin to increase the fraction of drug absorbed in stomach. MATERIALS AND METHODS: Sodium alginate and sodium carboxymethylcellulose were used to formulate the microsphere by ionotropic gelation with calcium chloride. The formulation was optimized using a three-factor, three-level Box-Behnken design. RESULTS: The particle size varied from 559.50 to 801.10 µm, entrapment efficiency from 61.29 to 81.00% and in vitro release from 69.40 to 83.70%. The optimized formulation was found using point-prediction, and formulation OF-3 showed optimum results at 608.21 µm size, 79.65% entrapment efficiency and 82.72% drug release and 81% mucoadhesion up to 10 h. The drug release was controlled for more than 12 h. DISCUSSION: The particle size was most influenced by sodium alginate while entrapment efficiency and drug release depended upon both polymers. The release followed Higuchi model. CONCLUSION: Gastroretentive formulation was successfully optimized by a three-factor, three-level Box-Behnken design and found to be useful.

Spectral, thermal and morphological studies of chromium nanoparticles.

Cr(0) nanoparticles were synthesized by solution reduction process successfully. The influence of parameters on the size of Cr(0) nanoparticles was studied and the referential process parameters were obtained. The morphology and structure of the synthesized Cr(0) nanoparticles were characterized by Transmission Electron Microscopy (TEM), Powder X-ray Diffraction (XRD), Thermal Gravimetric Analysis (TGA), QELS Data and Infrared Spectroscopy (IR). The results show that Cr(0) nanoparticles are of high purity. XRD analysis revealed all relevant Bragg's reflection for crystal structure of Cr metal. XRD spectrum also indicates that there is no oxidation of Cr(0) nanoparticles to chromium oxide. TEM showed nearly uniform distribution of particles in methanol which was confirmed by QELS. Cr(0) nanoparticles can be synthesized easily by reducing agent and are quite stable too.

Modulation of synthetic parameters of cobalt nanoparticles: TEM, EDS, spectral and thermal studies.

The study focuses on the modulation of synthetic parameters in order to influence the size, structure, composition and arrangement of nanoparticles of cobalt. Cobalt nanoparticles were synthesized by ethanolic solution of benzildiethylenetriamine in cobalt nitrate solution at 60 °C with stirring and refluxing leads to nanoparticles of cobalt. The morphology and structure of the synthesized nanoparticles of cobalt were characterized by Transmission Electron Microscopy (TEM), Powder X-ray Diffraction (XRD), Thermal Gravimetric Analysis (TGA), QELS Data and Infrared Spectroscopy (IR). Crystalline size was 20 nm determined from the sharp peak at 2θ=25 °C from the powder XRD. TEM images of cobalt nanoparticles without reducing agent having the diameter 20 nm with spherical shape and black color.

Modulation of synthetic parameters of novel zinc nanoparticles and reducing agent: powder X-ray diffraction, transmission electron microscopy and spectral studies.

Benzildiethylenetriamine was used as a reducing agent to prepare Zn nanostructures. The crystallography of benzildiethylenetriamine was done to confirm the molecular structure. These studies focus on the modulation of synthetic parameters in order to influence the size, structure, composition and arrangement of nanoparticles of Zinc. The morphology and structure of the synthesized nanoparticles of zinc were characterized by transmission electron microscopy (TEM), powder X-ray diffraction (XRD), energy dispersive spectrum analysis (EDS), thermal gravimetric analysis (TGA) and infrared spectroscopy (IR). The results show that nanoparticles of zinc are of high purity and 20 nm in size.

Synthesis of Al nanoparticles: transmission electron microscopy, thermal and spectral studies.

Nanoparticles of Al(0) were synthesized by solution reduction process successfully. The influence of parameters on the size of Al(0) nanoparticles were studied and the referential process parameters were obtained. The morphology and structure of the synthesized Al(0) nanoparticles were characterized by Transmission Electron Microscopy (TEM), Powder X-ray Diffraction (XRD), Thermal Gravimetric Analysis (TGA), QELS Data and Infrared Spectroscopy (IR). The results show that nanoparticles of Al(0) are of high purity. XRD analysis revealed all relevant Bragg's reflection for crystal structure of Al metal. XRD spectrum also indicates there is no oxidation of Al(0) nanoparticles to aluminum oxide. TEM showed nearly uniform distribution of the particles in methanol and it was confirmed by QELS. Al(0) nanoparticles can be synthesized easily by reducing agent and are quite stable too.

Prompt laboratory diagnosis in timely containment of a plague outbreak in India.

A focal outbreak of pneumonic plague occurred in a hamlet of village Hatkoti, district Shimla, Himachal Pradesh in the first fortnight of February, 2002. A total of 16 cases with 4 deaths were reported. Diagnosis of plague was confirmed by the laboratory in 10 (63%) cases. Y. pestis was isolated from clinical samples of 3 cases and confirmed by bacteriophage lysis. Molecular tests confirmed the presence of Y. pestis specific pla and F1 genes in 4 cases; DNA fingerprinting had identity with the known sequence of plague bacilli. Paired samples from 5 cases showed more than 4 fold rise and 1 case showed more than 4 fold fall in antibodies against F1 antigen of Y. pestis. The present communication emphasises that timely and systematic laboratory investigations give confirmatory diagnosis in shortest possible time which forms the backbone of the outbreak control in a timely fashion and prevents confusion and controversy.