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The physical and mechanical properties of German Angora rabbit fiber obtained from the Northeast region of India were studied. The fiber fineness, staple length, diameter, and medullation of the overall fleece of the rabbit were 0.22 ± 0.04 tex, 09.10 ± 2.21 cm, 23.5 ± 1.04 µm, and 85.46 ± 1.23%, respectively. The cross-sectional shape was found elliptical to oval, and the average tenacity, breaking elongation, specific work of rupture, and initial modulus were 24.78 ± 11.48 cN/tex, 28.78 ± 09.06%, 3.46 ± 2.31cN/tex, and 1065.13 ± 826.67 cN/tex, respectively. The whiteness index average ash and moisture content were 61.24 ± 2.32, 1.36 ± 0.12, and 10.43 ± 0.57%, respectively. Three blend ratios of Angora rabbit fiber and viscose fiber (50:50, 25:75, and 0:100) were used to make yarns having 55 tex, 86 tex, and 97 tex linear density from each blend. Tenacity and breaking elongation significantly (p =.00) increased with an increase in the rabbit fiber composition in the blend and was maximum at the 50:50 blend (7.79 ± 1.92 cN/tex and 7.64 ± 1.70%, respectively). Yarn diameter increased with the increase in yarn count. With the decrease in the rabbit fiber composition from 50% to 25% in the yarn, the friction coefficient increased and the abrasion resistance decreased significantly (p =.00). [ABSTRACT FROM AUTHOR]

Indian citrus ringspot virus (ICRSV), a member of the Mandarivirus genus, causes citrus ringspot disease, impacting kinnow orange quality and yield. Early and accurate detection methods are crucial before visible symptoms manifest in plants. In this study, a 507 bp partial coat protein gene (pCPG) segment was amplified from infected kinnow leaf tissues, cloned into a pET28a vector, and transformed into E. coli BL21(DE3) cells. Induced with IPTG, the cells overexpressed a recombinant partial coat protein (rpCP) of approximately 23 kDa, purified using Ni-NTA resin via affinity chromatography. Validated in western blot with an anti-His antibody, rpCP was used to generate an ICRSV-specific polyclonal antibody (PAb) in rabbits. PAb, optimized at 1:1000 dilution, successfully detected ICRSV in infected kinnow orange leaf extracts via DAC-ELISA and IC-RT-PCR assays. ICRSV was detectable in sample dilutions up to 1:640 and 1:10240 (w/v, g mL -1 ) by DAC-ELISA and IC-RT-PCR, respectively. One-step RT-PCR assays were also optimized, confirming the presence of ICRSV by amplifying a 507 bp pCPG fragment from total RNA extracted from kinnow orange leaves, with dilution up to 1:5120 (w/v, g mL -1 ). The result demonstrated that IC-RT-PCR has a 16-fold and 2-fold higher sensitivity than DAC-ELISA and one-step RT-PCR assays., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper, (Copyright © 2024 Elsevier B.V. All rights reserved.)

Elephant endotheliotropic herpesviruses (EEHV) belong to the family Herpesviridae and cause a highly fatal hemorrhagic infection in elephants. EEHV poses a global threat to the already endangered elephant population. Since EEHV is a non-cultivable virus, there is a scarcity of specific diagnostics, therapeutics, and vaccines. In this study, our objective was to develop biologicals for diagnosis and pathological studies against the most prevalent EEHV1A/1B. We expressed two truncated fragments of the DNA polymerase, glycoprotein B (gB), and glycoprotein (gL) of EEHV in the prokaryotic system. Hyperimmune serum against the purified antigens was raised in rabbits and guinea pigs. We validated the reactivity of this hyperimmune serum using western blotting, ELISA, and immune-histochemistry on known positive infected tissues. Samples collected from 270 animals across various states in India were evaluated with these biologicals. The raised antibodies successfully demonstrated virus in immune-cytochemistry. Additionally, all known positive samples consistently exhibited significant inhibition in the OD values when used in the competitive format of ELISA across all four antigens when compared to the serum collected from known negative animals. An apparent sero-prevalence of 10 % was observed in the randomly collected samples. In summary, our study successfully developed and validated biologicals that will be invaluable for EEHV diagnosis and control., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Introduction: The two formulations of antihuman thymocyte immunoglobulin that are used as T‑cell depleting induction agents in renal transplantation are anti‑thymocyte globulins(ATG) thymoglobulin and (antiT‑lymphocyte globulin [ATLG]‑Grafalon‑formerly ATG‑Fresenius). Very few trials have compared these two formulations. In this retrospective study, we compared the incidence of infections, rejections, graft survival, mortality, and lymphocyte profile of ATG and ATLG. Materials and Methods: This was a single‑center retrospective study of 127 consecutive kidney‑alone transplant recipients from January 2014 to June 2019. Patients received 3 mg/kg of ATG or 4 mg/kg single dose of ATLG. CD 3 counts were done on day 3 of the transplant. Most of the recipients received standard triple immunosuppression of tacrolimus, mycophenolate mofetil, and prednisolone. Results: Fifty‑eight patients received ATG and 69 patients received ATLG. Baseline demographics were similar between the two groups. Death‑censored graft survival(99%) (P = 0.258) and biopsy‑proven acute rejection (BPAR) (32% vs. 29%, P = 0.128) were similar in both groups. Graft survival was better in ATLG group (92.7% vs. 87.5% P = 0.020). Bacterial infections (41.1% vs. 27.6%, P = 0.03) and sepsis‑related mortality (11.54% vs. 4.34% P = 0.02) were significantly higher in the ATG group. Conclusion: ATLG, when used as an induction agent, was associated with a lesser rate of bacterial infections and sepsis‑related mortality, but better graft survival as compared to ATG but has comparable BPAR, death‑censored graft survival, and viral/fungal infections. [ABSTRACT FROM AUTHOR]

Domestic rabbit belongs to genus Oryctolagus cuniculus which has 38 important breeds and 77 varieties of European rabbits. The domestic rabbit is known to produce quality meat, fur and very fine quality animal fibre. Rabbit rearing has gained momentum in the recent years among the developing countries including India, owing to their small body size, rapid growth rate, high prolificacy, early maturity, shorter generation interval and ability to utilize forage and fibrous agricultural by-products. In India, there has been a rising awareness in recent years on the virtues of broiler rabbit production as an alternative means of alleviating food shortages. India is also one of the developing countries which are also facing meat shortage of 4.66 g/day against the recommended requirement of 87 g/day. In India, the total rabbit population has increased from 424 thousands in 2007 to 591.6 thousand in 2012 which is 39.55% increase in last 5 years. [ABSTRACT FROM AUTHOR]

The present study reports the clinico-pathologic changes observed in spontaneous hepatic coccidiosis caused by Eimeria stiedae in rabbits at Bannerghatta Biological Park (BBP) in Karnataka state of India. The clinical signs observed in the affected rabbits included sudden death, stunted growth, anorexia, weight loss, diarrhoea and abdominal pain. Increased liver enzymes and bilirubin was noticed upon biochemical examination. At necropsy, lesions included grossly enlarged liver (hepatomegaly) with presence of discrete yellowish-white nodules ranging from 1mm to 5mm size throughout the parenchyma containing a thick creamy white fluid. Faecal sample examination demonstrated the presence of Eimeria oocyts. The impression smears from the liver and intestinal mucosa also showed Eimeria oocysts, degenerative changes, necrotic cells and mononuclear cell aggregation to form oocyst granuloma. Histological observation revealed distension of bile duct, hyperplasia of epithelium along with numerous coccidian oocysts at the centre. Granulomatous tissues were observed to have encircled the bile duct with infiltration of inflammatory cells. Microscopic examination of exudates from liver also revealed numerous oocysts of E. stiedae. Multiple areas of coagulative necrosis of hepatic cells surrounded with inflammatory cells were also found. [ABSTRACT FROM AUTHOR]

Tuberose flowers (Calcutta Single variety) valued as ornamentals globally, have short shelf-lives of 8 days at 4 ± 1 °C and are therefore discarded post senescence. Previous investigations from our laboratory have established that a combination treatment using GRAS preservatives [(sucrose (4%) and CaCl 2 (0.02%)]-cum-gamma-irradiation (0.02 kGy) could extend its shelf-life to 24 days, when stored at 4 ± 1 °C with concomitant enhancement in the content of its bioactive principle, viz. methyl eugenol. Supercritical carbon dioxide (SC-CO 2 ) extract of the tuberose flower wastes post combination treatment therefore had a higher methyl eugenol content (4.11 ± 0.05 µg/g), vis-à-vis its non-treated counterpart (2.03 ± 0.03 µg/g), and thus significantly higher antioxidant and antimicrobial potencies (MIC values of 1.83 ± 0.02 mg/ml and 1.98 ± 0.03 mg/ml against S. aureus ATCC 25923 strain and MDR strain, respectively). The microencapsulated powder of the extract (ME p ) obtained by spray drying was applied for healing of epidermal wounds created on New Zealand white rabbits, post skin irritancy test (wherein no clinical sign of toxicity, redness or swelling was observed). When ME p was applied, accelerated healing occurred which commenced on day 2 and was completed by day 6 vis-à-vis that of the control powder set (without extract) which showed no signs of wound healing. Therefore, the sensorially compromised-senesced tuberose flowers, a rich source of methyl eugenol, has been successfully valorized through utilization of the same in developing a novel topical antibiotic powder against potent skin pathogens., (© 2023. The Author(s), under exclusive licence to Springer Nature B.V.)

Background and Purpose: Definitive diagnosis of Creutzfeldt–Jakob disease (CJD) requires demonstration of infective prion protein (PrPSc) in brain tissues by immunohistochemistry or immunoblot, making antemortem diagnosis of CJD difficult. The World Health Organization (WHO) recommends detection of 14-3-3 protein in cerebrospinal fluid (CSF) in cases of dementia, with clinical correlation, as a useful diagnostic marker for CJD, obviating the need for brain biopsy.This facility is currently available in only a few specialized centers in the West and no commercial kit is available for clinical diagnostic use in India. Hence the objective of this study was to develop an in-house sensitive assay for quantitation of 14-3-3 protein and to evaluate its diagnostic potential to detect 14-3-3 proteins in CSF as a biomarker in suspected cases of CJD. Materials and Methods: A minigene expressing the “core” 14-3-3 protein was synthesized by overlapping polymerase chain reaction (PCR) and the recombinant protein was produced by employing a bacterial expression system. Polyclonal antibodies raised in rabbit against the purified recombinant protein were used for developing a dot blot assay with avidin-biotin technology for signal amplification and quantitation of 14-3-3 protein in CSF. Results: The results in the present study suggest the diagnostic potential of the dot blot method with about 10-fold difference (P < 0.001) in the CSF levels of 14-3-3 protein between the CJD cases (N = 50) and disease controls (N = 70). The receiver operating characteristic (ROC) analysis of the results suggested an optimal cutoff value of 2 ng/mL. Conclusions: We have developed an indigenous, economical, and sensitive dot blot method for the quantitation of 14-3-3 protein in CSF. [ABSTRACT FROM AUTHOR]

Background: Poloxamer 407 is used in parenteral formulations as solubilizing and wetting agent for traditional, low-molecular-weight organic drug molecules and as stabilizing agent for proteins and polypeptide drugs. It has very high promising value in the medicine field, but has held responsible for changes in the lipid parameters. Thus, effect of poloxamer 407 on the rabbit's serum very low density lipoprotein (VLDL), low-density lipoprotein (LDL), and high-density lipoprotein (HDL) levels was studied after being injected intravenously. Aims & Objective: To study the effect of poloxamer 407 on rabbit's serum VLDL, LDL, and HDL levels following intravenous injection. Materials and Methods: Pretreatment, baseline readings were recorded. Rabbits were injected with 5.5, 27.5, and 137.5 mg/kg of poloxamer 407, and the effects on blood chemistry were assessed on the 2nd, 4th, and 7th day. The results of the study were expressed as mean ± SEM, and data were analyzed using oneway analysis of variance test. Values with P < 0.05 were considered as significant. Results: The highest dose of poloxamer 407 (137.5 mg/kg) significantly increased serum VLDL and decreased HDL level in rabbits, with the maximum increase observed on the 2nd day after injection. All the doses given did not alter the serum LDL level. The lower doses of poloxamer 407 did not alter serum VLDL, LDL, and HDL levels. Conclusion: Our results showed that poloxamer 407 in higher doses significantly increased serum VLDL and decreased HDL levels but in lower doses did not show any adverse effect on the lipid biochemistry. But we recommend further studies to know the effect of this poloxamer on chronic use and through different route of administration. [ABSTRACT FROM AUTHOR]

Abstract: Ethnopharmacological relevance: Corchorus depressus Linn. has been used as an aphrodisiac in traditional Indian medicine to treat male sexual dysfunction and impotency. Aim of the study: The petroleum ether, chloroform, ethyl acetate, n-butanol and aqueous fractions of 95% methanol extract of Corchorus depressus were screened initially for their in vitro aphrodisiac activity on rabbit corpus cavernosum smooth muscle. The chloroform fraction (CDC) was found to be the most active and therefore investigated further on general mating behavior, libido and potency of normal male Wistar albino rats in comparison with the standard drug, Sildenafil citrate. Materials and methods: Animals were divided into the following groups: Control, SC CDC 100, CDC 200, and CDC 400, who received saline, Sildenafil citrate or the chloroform fraction of Corchorus depressus at doses of 100, 200 or 400mg/kg b.wt., respectively. The route of administration for all the groups was oral dosing, which was once in a day for 45 days. To analyze the mating behavior, female rats with estrus phase were used. Results: The chloroform fraction of methanolic extract of Corchorus depressus significantly reduced ML, IL, PEI and III. There was a significant increase in the MF, IF and EL and serum testosterone levels throughout the study period. The potency test significantly increased erections, quick flips, long flips and total reflex. In vitro aphrodisiac activity was significantly higher in chloroform fraction at a concentration of 25.0mg/ml, which induced 71.4% relaxation. The combined results of the above mentioned models indicate that the chloroform fraction of Corchorus depressus produces a significant increase in sexual activity as exhibited by 25mg/ml in vitro and 400mg/kg in vivo. In comparison with the control, all the drug-treated groups have shown drug-induced effects for a few parameters. Conclusions: In vitro and in vivo studies provide valuable experimental evidence that the chloroform fraction of methanolic extract of Corchorus depressus possesses aphrodisiac property. This study further substantiates the ethnopharmacological claims of Corchorus depressus as a sexual stimulating agent and offers a significant potential for studying the effect on male sexual response and its dysfunctions. [Copyright &y& Elsevier]

As an alternative to broiler chicken, rearing rabbits for meat and fur skins is gaining momentum now a days because of the fact that the meat from rabbit is low in cholesterol and the most delicious. Rabbits were reared as laboratory animals till 1970. Commercial aspects of rabbit rearing started after the import of exotic rabbits during 1970s by CSWRI, Indian Council of Agricultural Research for the first time in our country. Rabbits are the only 'Micro livestock' suitable for rearing by small farmers and landless agricultural laborers with simple housing in their backyards and can be fed with forages, agricultural by-products and kitchen wastes. In fact backyard rabbitries are best suited for our country to increase the per capita income and per capita animal protein availability. [ABSTRACT FROM AUTHOR]

The tomato spotted wilt virus (TSWV) belonging to the genus Orthotospovirus, family Tospoviridae, causes severe necrotic disease in field crops and horticultural crops, resulting in considerable yield loss worldwide. The development of protein-based diagnostics is essential to track the virus transmission and prevent its spread in vegetatively propagated crops such as ornamentals. In this study, nucleocapsid (N) gene of TSWV was cloned in pET 28 a (+) expression vector. Expression of the 32 kDa recombinant TSWV-N protein was induced in BL21 (DE3) cells using 1 mM of Isopropyl β-d-1-thiogalactopyranoside (IPTG), and was confirmed through SDS-PAGE and Western blot by fluorescent-labeled secondary antibody. The bacterial cells expressed recombinant TSWV-N protein up to a concentration of 9.48 μg/mL. The purified protein was used for immunization of a rabbit to produce specific polyclonal antiserum. The TSWV antiserum was conjugated with the enzyme alkaline phosphatase (ALP). Double Antibody Sandwich-Enzyme Linked Immunosorbent Assay (DAS-ELISA) was developed and validated against TSWV infected hosts. This antiserum specifically reacted with recombinant N protein as well as TSWV infected hosts, but not with groundnut bud necrosis orthotospovirus (GBNV) as well as capsicum chlorosis orthotospovirus (CaCV) infecting tomato and chilli plants. The coating antibody at 1 μg/mL concentration and 1:500 dilution of enzyme conjugate were found to be effective and economical in the detection of recombinant N protein of TSWV and the virus present naturally in the infected hosts. Using standardized DAS-ELISA protocol, the TSWV titer also was quantified in artificially inoculated assay hosts. Among 11 hosts tested, higher virus titer was recorded in Nicotiana tabacum (0.270 μg/100 μL), followed by Impatiens balsamiana (0.185 μg/100 μL) and Dahlia pinnata at a low virus tire of 0.083 μg/100 μL. The diagnostic reagents and protocol (DAS-ELISA) are further validated by detecting the infection of TSWV in chrysanthemum stem cuttings from six different nurseries in the hill stations of Tamil Nadu, India. The DAS-ELISA assay experimented on six varieties from four different nurseries revealed that the Mum Yellow variety had a higher percentage of TSWV infection (36 %), which was followed by the Mum White variety (33 %); both collected from Kotagiri Nursery. The same variety exhibited a higher virus titer by DAS-ELISA, an A 405 value range of 0.733 (̴ 0.115 μg) and 0.711 (̴ 0.111 μg) respectively, and a total of 27 % of TSWV infection was confirmed by screening 800 stem cuttings by DAS-ELISA. The presence of TSWV was also detected in 54 (6.75 %) asymptomatic stem cuttings from different locations, and the A 405 value ranged from 0.325 to 0.468. (̴ 0.044-0.069 μg/100 μL); this is the first reported development of immune-based diagnostics for TSWV in India. This protocol and diagnostics will be highly useful for quarantine purposes while trading large quantities of planting materials., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Around 374 data on different mortality cases recorded from New Zealand White and Soviet Chinchilla breed of rabbit in three seasons i e summer, rainy and winter over a period of four years in Rabbit Research Farm of the institute under Animal Production Division were analyzed statistically. Rabbits were housed in indoor cage system and 50% mash feeding and 50% roughage feeding was practiced. It was found that highest mortality was in I st generation crops. Breed wise higher mortality was observed in New Zealand White breed of rabbit (22.12%). Sex wise higher mortality was in female (20.48%). Age wise mortality revealed that highest mortality was in finisher (21.49%). Season wise highest mortality was in summer (17.90%). Etiology of disease indicated that highest mortality was due to coccidiosis (5.62%) followed by deficiency disorder (5.50%). In 1 st and 2 nd year there was significant differences (P<0.05) in mortality among grower, finisher and adult rabbits. There were also significant (P<0.05) differences in mortality between different seasons in 1 st year. There was significant difference of mortality between the generations in 1 st year. In 2 nd year breed difference was observed to be significant. [ABSTRACT FROM AUTHOR]

Economics of rearing two meat type rabbits in hot humid conditions of West Bengal was undertaken. The animals were maintained in galvanized iron wire cages under routine farm feeding and health care management. Net annual return per doe was estimated with an aim to compare efficiency of the two breeds. Four crops were produced per year in both the breeds. The total numbers of kits born and survived up to the age of slaughter in a year were more in Grey Giant than Soviet Chinchilla. The individual live weight at slaughter was slightly more in Soviet Chinchilla than Grey Giant, but total weight of surviving youngs at this stage was more in Grey Giant than Soviet Chinchilla. This might be due to more number of kits born and survived at this stage in Grey Giant rabbits. Considering the selling price of rabbits on live weight basis and cost of feeds involved, an amount of Rs.221.25 may be expected from a Soviet Chinchilla doe and Rs.295.00 from a Grey Giant doe during a period of one year indicating better economic performance of the latter breed. The rabbit husbandry can be successfully advocated especially for the poorer section of the society for generating their subsidiary income and improving family nutrition status as it calls for low input and easy husbandry practices. [ABSTRACT FROM AUTHOR]

Vibrio cholerae, causative agent of the water-borne disease cholera still threatens a large proportion of world's population. The major biotypes of the pathogen are classical and El Tor. There have been recent reports of variant V. cholerae strains circulating around the world. In the present study, the epidemiological status of V. cholerae strains circulating in the country over a decade was assessed. Also, a comprehensive analysis of the difference in pathogenicity between the different biotypes of V. cholerae strains was evaluated both in-vitro and in-vivo. The amount of CT produced by different biotypes of V. cholerae strains were analyzed by GM 1 ELISA and the probable reasons for the difference in toxin production was discussed. MLST analysis grouped the isolates into a single Sequence Type (ST 69) whereas PFGE analysis clustered the isolates into ten different pulsotypes revealing molecular diversity. The circulating strains were identified to produce cholera toxin and CT mRNA intermediate to the classical and prototype El Tor strains. Also, the circulating strains were identified to possess four ToxR binding sequences. In-vivo pathogenicity analysis by rabbit ileal loop fluid accumulation assay revealed the Haitian variant strains to be more hyperemic than the prototype strains., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Background & Objectives: Due to the emergence of Kyasanur forest disease (KFD) virus to new regions in India, there is an urgent need to develop an early diagnostic system, which is cost-effective and can be efficiently used with minimum paraphernalia. The non-structural-1 (NS1) protein is known to be an early diagnostic marker for flaviviruses. Furthermore, NS1 antigen capture ELISA kits developed using bacterially expressed dengue NS1 protein are commercially available., Methods: Based on the data available on dengue virus, West Nile virus and other flaviviruses, bacterially expressed Kyasanur forest disease virus (KFDV) NS1 protein and polyclonal serum raised against the NS1 protein in mice and rabbit were used to develop an antigen capture ELISA for early diagnosis of the virus. The feasibility of this ELISA was further tested using in silico predictions., Results: KFDV NS1 gene was cloned, expressed and confirmed by SDS-PAGE and western blotting. An antigen detection ELISA was standardized and sensitivity and specificity was tested with other flaviviruses. KFDV acute phase 43 samples were tested and only two were found to be positive for KFDV NS1 antigen. Superimposition of KFDV NS1 and TBEV NS1 revealed a root mean square distance (RMSD) of ~0.79 Å covering 1220 backbone atoms. This implies that the structures are very similar in terms of 3D fold. The identity of amino acid composition between these proteins was 73.4% and similarity was 92.9%, as revealed from the pairwise comparison., Interpretation & Conclusion: The study points out that the half-life, expression and secretion levels of KFDV NS1 protein are not sufficient enough for its use as early diagnostic marker. The protein may have to be expressed in eukaryotic host to counter the lack of glycosylation in bacterial plasmid based expression of proteins. Hence, bacterially expressed KFDV NS1 protein may not be an ideal early diagnostic marker for the virus., Competing Interests: None

A rapid multiplex silver enhanced 'sandwich vertical flow immunogold assay (SVIA)' based on disposable porous filter-membrane was developed for on-site detection of oxytetracycline (OTC) residues in fish tissues. Artificial antigens were synthesized to raise selective anti-OTC rabbit antiserum and highly specific monoclonal antibody (mAb). The assay consists of three layers. The first layer was composed of immobilized anti-OTC rabbit antiserum (capture antibody) onto the membrane, the second layer was the molecule of interest (OTC) and the third layer was refined with detector immunogold labeled mAb. The sensitivity of the silver enhanced SVIA was as low as 2 ng mL -1 (within 4 min) representing a 125-fold increase in sensitivity over the HRP labeled sandwich vertical flow assay. The reliability of developed technique was co-evaluated with HPLC and validated using 115 field fish samples from different regions of southern India. The results demonstrated that SVIA has the high potentiality to be established as a semi-quantitative routine on-site screening tool., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Fowl adenovirus-4 (FAdV-4) causes hydropericardium syndrome (HPS) in poultry worldwide. An understanding of viral structural protein composition is important for developing novel immunodiagnostics and immunoprophylactics. Here we report isolation, culture, molecular and protein profile of FAdV-4 isolates recovered from HPS outbreaks in chicken in the states of Himachal Pradesh and Tamil Nadu in India. We performed a sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting-based protein profiling of FAdV-4 isolates against a reference FAdV-1 or Chicken Embryo Lethal Orphan (CELO) virus. SDS-PAGE analysis showed that seven protein bands in FAdV-4 isolates were similar to CELO expect an additional band of 110 kDa in CELO virus. On Western blotting, two protein fractions of 43 kDa and 78 kDa size were observed in FAdV-4 isolates. Overall, results show that FAdV-4 isolates recovered from different regions of the country had similar protein profile and possibly a common source of origin., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

India remained an epicenter for the snakebite-related mortality and morbidities due to widespread agricultural activities across the country and a considerable number of snakebites offended by Indian cobra (Naja naja), common krait (Bungarus caeruleus), Russell's viper (Daboia russelii), and saw-scaled viper (Echis carinatus). Presently, there is no selective test available for the detection of snake envenomation in India before the administration of snake antivenin. Therefore, the present study aimed to develop rapid, sensitive assay for the management of snakebite, which can detect venom, responsible snake species and serve as a tool for the reasonable administration of snake antivenin, which have scarcity across the world. The selective envenomation detection assay needs venom specific antibodies (VSAbs) for that monovalent antisera was prepared by hyperimmunization of rabbits with specific venom. However, obtained antibodies exhibit maximum activity towards homologous venom as well as quantifiable degree of cross-reactivity with heterologous venoms. Use of these antibodies for development of selective envenomation detection assay may create ambiguity in results, therefore needs to isolate VSAbs from monovalent antisera. The cross-reacting antibodies were specifically removed by immunoaffinity chromatography to obtain VSAbs. For the development of venom detection ELISA test (VDET), two different species of antibodies were used that offers enhanced sensitivity along with selective identification of the venoms of the responsible snakes. In conclusion, the developed VDET is rapid, specific, yet sensitive to detect venoms of offending snake species, and its venom concentration down to 1.0 ng/ml. However, the device observed with lowest venom concentration detection ability in the range <1.0 ng/ml from experimentally envenomated samples. The implementation of VDET will help in avoiding unnecessary usage and adverse reactions of snake antivenin. The test has all the merits to become a choice of method in envenomation diagnosis from medically important snakes of India., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Pasteurella multocida causes haemorrhagic septicemia in livestock and wild animals, including elephants. The disease has been reported in Asian elephants in India and Sri Lanka, but to date there have been no reported cases in Thailand. ELISA or indirect hemagglutination assays (IHA) have been demonstrated to be able to detect the antibody against the disease in cattle, but no data are available for elephants. The present study reports a novel in-house indirect ELISA for antibody detection of haemorrhagic septicemia in Asian elephants, and evaluates the sensitivity and specificity of the method using a Bayesian approach. The characteristics of ELISA and IHA were analyzed using a one population Bayesian model assuming conditional dependence between these two diagnostic tests. The IHA was performed as recommended by the World Organization for Animal Health (OIE) manual for haemorrhagic septicemia. An in-house indirect ELISA was developed with a heat extract antigen of P. multocida strain M-1404 (serovar B:2) as a coating antigen and rabbit anti-immunoglobulin G conjugated with horseradish peroxidase (eIgG-HRP). The checkerboard titration method was done using elephant sera immunized with P. multocida bacterin and negative sera from colostrum-deprived elephant calves. The concentrations of heat extract antigen (160μg/ml), sample serum (1:100), and eIgG-HRP (1:1000) were optimal for the assay. The calculated cut-off value was 0.103. Of the elephant sera, 50.59% (43/85) were considered seropositive by ELISA. The sensitivity of the ELISA test was higher than that of the IHA test [median=86.5%, 95% posterior probability interval (PPI)=52.5-98.9%] while the specificity was lower (median=54.1%, PPI=43.6-64.7%). The median sensitivity and specificity of IHA were 80.5% (PPI=43.8-98.0%) and 78.4% (PPI=69.0-87.0%), respectively. These findings suggest that our in-house indirect ELISA can be used as a tool to detect the antibody against haemorrhagic septicemia in Asian elephants., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Rabies a fatal viral zoonosis is endemic in India. There is no report on phylogenetic study of Indian rabies virus isolates based on the complete G gene. In the present study, a total of 25 rabies positive brain samples collected during 2001-2014 from North India (UP, MP, Delhi, Rajasthan), South India (Kerala and Karnataka) and Gujarat states belonging to six different host species were subjected to G gene amplification by RT-PCR as three overlapping fragments of 881 bp, 991 bp and 618 bp. Phylogenetic analysis revealed that all Indian rabies virus isolates are genetically closely related with Arctic-like 1a lineage viruses. However, two distinct clusters were identified namely, India South and India North. All the Indian rabies isolates had 95.5-100% homology related to geography, but not to host species. Deduced amino acids on comparison revealed two amino acid changes, aa 356 in ECTO; N→K and aa 458; M→I, which were found to distinguish between the India South and India North isolates., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Background: Mass vaccination campaigns of the population aged 1-29 years with 1 dose of group A meningococcal (MenA) conjugate vaccine (PsA-TT, MenAfriVac) in African meningitis belt countries has resulted in the near-disappearance of MenA. The vaccine was tested in clinical trials in Africa and in India and found to be safe and highly immunogenic compared with the group A component of the licensed quadrivalent polysaccharide vaccine (PsACWY). Antibody persistence in Africa and in India was investigated., Methods: A total of 900 subjects aged 2-29 years were followed up for 4 years in Senegal, Mali, and The Gambia (study A). A total of 340 subjects aged 2-10 years were followed up for 1 year in India (study B). In study A, subjects were randomized in a 2:1 ratio, and in study B a 1:1 ratio to receive either PsA-TT or PsACWY. Immunogenicity was evaluated by measuring MenA serum bactericidal antibody (SBA) with rabbit complement and by a group A-specific immunoglobulin G (IgG) enzyme-linked immunosorbent assay., Results: In both studies, substantial SBA decay was observed at 6 months postvaccination in both vaccine groups, although more marked in the PsACWY group. At 1 year and 4 years (only for study A) postvaccination, SBA titers were relatively sustained in the PsA-TT group, whereas a slight increasing trend, more pronounced among the youngest, was observed in the participants aged <18 years in the PsACWY groups. The SBA titers were significantly higher in the PsA-TT group than in the PsACWY group at any time point, and the majority of subjects in the PsA-TT group had SBA titers ≥128 and group A-specific IgG concentrations ≥2 µg/mL at any point in time in both the African and Indian study populations., Conclusions: Four years after vaccination with a single dose of PsA-TT vaccine in Africa, most subjects are considered protected from MenA disease., Clinical Trials Registration: PsA-TT-003 (ISRCTN87739946); PsA-TT-003a (ISRCTN46335400)., (© The Author 2015. Published by Oxford University Press on behalf of the Infectious Diseases Society of America.)

A bovine rotavirus pentavalent vaccine (BRV-PV) containing rotavirus human-bovine (UK) reassortant strains of serotype G1, G2, G3, G4 and G9 has been developed by the Serum Institute of India Ltd, in collaboration with the National Institute of Allergy and Infectious Diseases (NIAID), USA. The vaccine underwent animal toxicity studies and Phase I and II studies in adults, toddlers and infants. It has been found safe and immunogenic and will undergo a large Phase III study to assess efficacy against severe rotavirus gastroenteritis., (Copyright © 2014. Published by Elsevier Ltd.)

Background: Chikungunya virus (CHIKV) has emerged as one of the most important arboviruses of public health significance in the past decade. The virus is mainly maintained through human-mosquito-human cycle. Other routes of transmission and the mechanism of maintenance of the virus in nature are not clearly known. Vertical transmission may be a mechanism of sustaining the virus during inter-epidemic periods. Laboratory experiments were conducted to determine whether Aedes aegypti, a principal vector, is capable of vertically transmitting CHIKV or not., Methodology/principal Findings: Female Ae. aegypti were orally infected with a novel ECSA genotype of CHIKV in the 2nd gonotrophic cycle. On day 10 post infection, a non-infectious blood meal was provided to obtain another cycle of eggs. Larvae and adults developed from the eggs obtained following both infectious and non-infectious blood meal were tested for the presence of CHIKV specific RNA through real time RT-PCR. The results revealed that the larvae and adults developed from eggs derived from the infectious blood meal (2nd gonotrophic cycle) were negative for CHIKV RNA. However, the larvae and adults developed after subsequent non-infectious blood meal (3rd gonotrophic cycle) were positive with minimum filial infection rates of 28.2 (1∶35.5) and 20.2 (1∶49.5) respectively., Conclusion/significance: This study is the first to confirm experimental vertical transmission of emerging novel ECSA genotype of CHIKV in Ae. aegypti from India, indicating the possibilities of occurrence of this phenomenon in nature. This evidence may have important consequence for survival of CHIKV during adverse climatic conditions and inter-epidemic periods.

The venom of the largest venomous snake, the king cobra (Ophiophagus hannah), is still out of league for the production of therapeutic polyvalent antivenom nor it is characterized immunologically in the Indian subcontinent. In the present study, the king cobra venom is comparatively studied for the cross-reactivity/reactivity and toxicity neutralization by the locally available equine therapeutic polyvalent BSV and VB antivenoms, and monovalent antivenom (OH-IgG) prepared in rabbit. None of the two therapeutic antivenoms procured from two different firms showed any signs of cross-reactivity in terms of antigen-antibody precipitin lines in immunodouble diffusion assay; however, a weak and an insignificant cross-reactivity pattern was observed in ELISA and Western blot studies. Further, both BSV and VB antivenoms failed to neutralize proteolytic, hyaluronidase and phospholipase activities as well as toxic properties such as edema, myotoxicity and lethality of the venom. As expected, OH-IgG showed strong reactivity in immunodouble diffusion, ELISA and in Western blot analysis and also neutralized both enzyme activities as well as the toxic properties of the venom. Thus, the study provides insight into the likely measures that are to be taken in cases of accidental king cobra bites for which the Indian subcontinent is still not prepared for., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Several limitations of the use of embryonated eggs and the threat of pandemics have highlighted the need for other platforms for the production of influenza vaccines. We report the indigenous development and pre-clinical testing of an MDCK-based H1N1 pandemic influenza vaccine HNVAC from India. The cell bank and virus seed were characterized extensively. The cells were characterized by PCR, electron microscopy, and karyotyping, and found to be of female canine epithelial origin. The virus was confirmed by neutralization, haemagglutination inhibition, neuraminidase inhibition, and PCR and nucleotide sequencing. Adventitious agent testing was performed by both in vitro and in vivo studies. The in vitro studies included culturing, haemadsorption, haemagglutination, PCR and RT-PCR, whereas in vivo studies included passage in embryonated eggs and in laboratory animals. Both cell bank and virus seed were free of adventitious agents. MDCK cell lysates as well as cellular DNA did not produce tumours in newborn or adult laboratory animals. The bioprocess parameters were standardized to recover antigen with minimal levels of process-related impurities. The vaccine bulk was tested for the presence of specific antigen, and quantified by single radial immunodiffusion. Finally, non-adjuvanted and aluminium hydroxide adjuvanted vaccine formulations were found to be safe in preclinical toxicity studies in mice, rats, guinea pigs and rabbits, and immunogenic in mice and rabbits. This is the first and only cell culture-based influenza vaccine platform developed in any developing country., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Background & Objectives: Despite major control efforts, malaria remains a major public health problem that still causes high mortality rate worldwide especially in Africa and Asia. Accurate and confirmatory diagnosis before treatment initiation is the only way to control the disease. The present study was undertaken to develop reagents using sandwich ELISA for simultaneous detection of PfHRP2 (Plasmodium falciparum histidine rich protein) and PfLDH (P. falciparum lactate dehydrogenase) antigens in the proven malaria cases., Methods: The antibodies were raised against two epitopes of PfHRP2 protein and three unique and unexplored epitopes of PfLDH protein. These antibodies were able to detect PfHRP2 and PfLDH antigens in culture supernatant and parasitized RBC lysate of P. falciparum, respectively up to 50 parasites/μl. The in-house reagents were tested in 200 P. falciparum positive patients residing in Baghpat district of Uttar Pradesh in northern India., Results: Microsphere (PLGA) with CpG ODN were used to generate high titre and high affinity antibodies against selected peptides of PfHRP-2 and pLDH antigen in mice and rabbit. The peptide specific peak titre varied from 12,800 - 102,400 with an affinity ranging 0.73 - 3.0 mM. The indigenously developed reagents are able to detect PfHRP2 and PfLDH antigens as low as 75 parasites/μl of blood with a very high sensitivity (96-100%) and specificity (100%)., Interpretation & Conclusions: The study highlight the identification of unique epitopes of PfHRP2 and PfLDH, and the generated antibodies against these antigens were used for quantitative estimation of these two antigens using sandwich ELISA. No corresreactivity with P. vivax infected patients was observed with the sera.

Background: Vibrio cholerae non-O1/ non-O139 serogroups have been reported to cause sporadic diarrhoea in humans. Cholera toxins have been mostly implicated for hypersecretion of ions and water into the small intestine. Though most of the V. cholerae non-O1/ non-O139 strains lack these cholera toxins, several other innate virulence factors contribute towards their pathogenicity. The environmental isolates may thus act as reservoirs for potential spreading of these virulence genes in the natural environment which may cause the emergence of epidemic-causing organisms., Results: The environmental isolates of vibrios were obtained from water samples, zooplanktons and phytoplanktons, from a village pond in Gandhinagar, Gujarat, India. They were confirmed as Vibrio cholerae non-O1/ non-O139 using standard biochemical and serotyping tests. PCR experiments revealed that the isolates lacked ctxA, ctxB, tcpA, zot and ace genes whereas other pathogenicity genes like toxR, rtxC, hlyA, hapA and prtV were detected in these isolates. Compared with epidemic strain V. cholerae O1 El Tor N16961, culture supernatants from most of these isolates caused higher cytotoxicity to HT29 cells and higher hemolytic, hemagglutinin and protease activities. In rabbit ileal loop assays, the environmental isolates showed only 2-4 folds lesser fluid accumulation in comparison to N16961 and a V. cholerae clinical isolate IDH02365 of 2009. Pulsed Field Gel electrophoresis and Random amplification of Polymorphic DNA indicated that these isolates showed considerable diversity and did not share the same clonal lineage even though they were derived from the same water source. All the isolates showed resistance to one or more antibiotics., Conclusion: Though these environmental isolates lacked the cholera toxins, they seem to have adopted other survival strategies by optimally utilising a diverse array of several other toxins. The current findings indicate the possibility that these isolates could cause some gastroenteric inflammation when ingested and may serve as progenitors for overt disease-causing organisms.

The present work was aimed to develop an antiseptic cream formulation of Indian basil oil utilizing hydrophilic-lipophilic balance approach. In order to determine the required-hydrophilic lipophilic balance (rHLB) of basil oil, emulsions of basil oil were prepared by phase inversion temperature technique using water, Tween 80, and Span 80. Formulated emulsions were assessed for creaming (BE9; 9.8, BE10; 10.2), droplet size (BE18; 3.22 ± 0.09 μ m), and turbidity (BE18; 86.12 ± 2.1%). To ensure correctness of the applied methodology, rHLB of light liquid paraffin was also determined. After rHLB determination, basil oil creams were prepared with two different combinations of surfactants, namely, GMS : Tween 80 (1 : 3.45) and SLS : GMS (1 : 3.68), and evaluated for in vitro antimicrobial activity, skin irritation test, viscosity and consistency. The rHLB of basil oil and light liquid paraffin were found to be 13.36 ± 0.36 and 11.5 ± 0.35, respectively. Viscosity, and consistency parameters of cream was found to be consistent over 90 days. Cream formulations showed net zone of growth inhibition in the range of 5.0-11.3 mm against bacteria and 4.3-7.6 mm against fungi. Primary irritation index was found to be between 0.38 and1.05. Conclusively stable, consistent, non-irritant, enriched antiseptic basil oil cream formulations were developed utilizing HLB approach.

Forskolin is the first pharmaceutical drug and product derived from a plant to be approved in India by the DCGI in 2006. Forskolin (7beta-acetoxy-8, 13-epoxy-1a, 6β, 9a-trihydroxy-labd-14-en-11-one) is a diterpenoid isolated from plant Coleus forskohlii (Lamiaceae). It is a lipid-soluble compound that can penetrate cell membranes and stimulates the enzyme adenylate cyclase which, in turn, stimulates ciliary epithelium to activate cyclic adenosine monophosphate, which decreases intraocular pressure (IOP) by reducing aqueous humor inflow. The topical application of forskolin is capable of reducing IOP in rabbits, monkeys, and humans. In its drug interactions, forskolin may act synergistically with epinephrine, ephedrine and pseudoephedrine. Whereas the effects of anti-clotting medications like warfarin, clopidogre, aspirin, anoxaparin, etc., may be enhanced by forskolin. Forskolin is contraindicated in the medications for people with ulcers as forskolin may increase acid level. Forskolin has a very good shelf-life of five years. Recently, its Ophthalmic inserts and in situ gels for sustained and delayed-release drug delivery systems were tested in New Zealand Albino Rabbits for its antiglaucoma efficacy. This drug review explains Forskolin as a drug, its antiglaucoma potential and recent findings of forskolin as an antiglaucoma agent. The literature search method used for this review was different databases and search engines like PubMed, International Pharmaceutical Abstracts, Google, Medicinal and Aromatic Plants (MAPA).

Envenomation by hump-nosed pit viper (Hypnale hypnale, Hh) in Sri Lanka has caused significant morbidity and mortality, attributed to 35% of total venomous snakebites. In Southwestern India (Kerala), H. hypnale was increasingly identified as a dangerous and common source of envenomation, second to the Russell's viper but ahead of the cobra bites. Unfortunately, there is still no specific antivenom to date. This study aims to investigate the immunological properties of the venom and to assess the feasibility of specific Hh antivenom production as well as the development of a diagnostic assay. Hh venom elicited satisfactory titers of anti-Hh IgG in rabbits after 3rd immunization. The anti-Hh IgG, isolated with caprylic acid precipitation method, was effective in neutralizing the venom lethality (potency=48 LD(50) per ml IgG) as well as its procoagulant, hemorrhagic and necrotic effects, indicating the possibility to produce the specific antivenom using the common immunization regime. Cross-reactivity studies using indirect ELISA showed that anti-Hh IgG cross-reacted extensively with several Asiatic crotalid venoms, particularly that of Calloselasma rhodostoma (73.6%), presumably due to the presence of venom antigens common to both snakes. Levels of immunological cross-reactivity were vastly reduced with double-sandwich ELISA. Further work demonstrated that the assay was able to distinguish and quantify venoms of H. hypnale, Daboia russelii and Echis carinatus sinhaleyus (three common local viperid) used to spike human sera at various concentrations. The assay hence may be a useful investigating tool for diagnosing biting species and studying the time course profile of venom concentrations in blood., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Introduction: Radioactive contamination can occur as a result of accidental or intentional release of radioactive materials (RM) into the environment. RM may deposit on clothing, skin, or hair. Decontamination of contaminated persons should be done as soon as possible to minimize the deleterious health effects of radiation. The goal of this study was to evaluate the decontamination efficiency (for residual contaminant) of the active components of "Shudhika," an indigenously developed skin decontamination kit. The study kit is for external radioactive decontamination of intact skin., Methods: Decontamination efficiency was evaluated on the skin surface of rabbit (n = 6) and human volunteers (n = 13). 99mTc sodium pertechnetate (200-250 μCi) was used as the radio-contaminant. Skin surface area (5 × 5 cm2) of thoracic abdominal region of the rabbit and the forearm and the palm of human volunteers were used for the study. Decontamination was performed by using cotton swabs soaked with chemical decontamination agents of the kit., Results: Decontamination efficiency (% of the contaminant removed) was calculated for each component of the study. Overall effectiveness of the kit was calculated to be 85% ± 5% in animal and 92% ± 3% in human skin surfaces. Running water and liquid soap with water was able to decontaminate volunteers' hand and animal skin up to 70% ± 5%. Chemical decontamination agents were applied only for trace residues (30% ± 5%). Efficiency of all the kit components was found up to be 20% ± 3% (animal) and 28% ± 2 (human), respectively. Residual contamination after final decontamination attempt for both the models was observed to be 12% ± 3% and 5% ± 2%. After 24 and 48 hours of the decontamination procedure, skin was found to be normal (no redness, erythema and edema were observed)., Conclusion: Decontaminants of the study kit were effective in removal of localized radioactive skin contamination when water is ineffective for further decontamination. By using the chemical decontaminants of the study kit, the use of water and radioactive waste generation could be reduced. Cross-contamination could also be avoided. During radiologic emergencies where water may be radioactively contaminated, the study kit could be used.

Animal experiments continue to play an integral role in Indian undergraduate medical education, even though alternatives are becoming increasingly available. In this context, this study aimed to assess the perceptions of pharmacology faculty members from medical colleges in southern India regarding the use of animals and alternatives in experimental pharmacology, and to determine the association between these perceptions and the socio-demographic characteristics of the participants. Data were collected from 59 faculty members of 15 medical colleges in southern India. The response rate was 84.3%. A 30-statement, five-domain questionnaire was used, with a global score of 120. The mean ± SD global score was 60.9 ± 17.3. Significant differences were observed in domain scores and individual statement scores with respect to the extent of teaching experience. There were no statistically significant differences in perceptions with respect to age, gender or educational qualifications. All the participating colleges were conducting at least 3-8 animal experiments per year on the rabbit, rat, mouse and frog/toad. The pharmacology faculty members in the southern India medical colleges included in the study (especially the more experienced teachers) supported animal use in undergraduate medical education, in spite of being aware of the drawbacks of animal experiments and the availability of alternatives.

Trypanosoma evansi is a causative agent of 'surra', a common haemoprotozoan disease of livestock in India causing high morbidity and mortality in disease endemic areas. The proteinases released by live and dead trypanosomes entail immunosuppression in the infected host, which immensely contribute in disease pathogenesis. Cysteine proteinases are identified in the infectious cycle of trypanosomes such as cruzain from Trypanosoma cruzi, rhodesain or brucipain from Trypanosoma brucei rhodesiense and congopain from Trypanosoma congelense. These enzymes localised in lysosome-like organelles, flagellar pocket and on cell surface, which play a critical role in the life cycle of protozoan parasites, viz. in host invasion, nutrition and alteration of the host immune response. The paper describes the identification of cysteine proteinases of T. evansi lysate, activity profile at different pH optima and inhibition pattern using a specific inhibitor, besides the polypeptide profile of an antigen. Eight proteinases of T. evansi were identified in the molecular weight (MW) ranges of 28-170 kDa using gelatin substrate-polyacrylamide gel electrophoresis (GS-PAGE), and of these proteinases, six were cysteine proteinases, as they were inhibited by L-3-carboxy-2,3-transepoxypropionyl-lecuylamido (4-guanidino)-butane (E-64), a specific inhibitor. These proteolytic enzymes were most reactive in acidic pH between 3.0 and 5.5 in the presence of dithiothreitol and completely inactive at alkaline pH 10.0. Similarly, the GS-PAGE profile of the serum samples of rats infected with T. evansi revealed strong proteolytic activity only at the 28-kDa zone at pH 5.5, while no proteolytic activity was observed in serum samples of uninfected rats. Further, the other zones of clearance, which were evident in T. evansi antigen zymogram, could not be observed in the serum samples of rats infected with T. evansi. The polypeptide pattern of the whole cell lysate antigen revealed 12-15 polypeptide bands ranging from 28 to 81 kDa along with five predominant polypeptides bands (MW of 81, 66, 62, 55 and 45 kDa), which were immunoreactive with hyperimmune serum (HIS) and serum of experimentally infected rabbits with T. evansi infection. The immunoblot recognised antibodies in experimentally infected rabbits and against HIS as well, corresponding to the zone of clearances at lower MW ranges (28-41 kDa), which may be attributed to the potential of these proteinases in the diagnosis of T. evansi infection. Since these thiol-dependent enzymes are most active in acidic pH and considering their inhibition characteristics, these data suggest that they resemble to the mammalian lysosomal cathepsin B and L.

Background: Vibrio spp., being primarily inhabitants of the aquatic environment, pose a severe health threat to humans. This problem is escalated in developing countries where water-logging after rainfall is very common. Therefore, screening of environmental water samples for the presence of clinically important species of Vibrio becomes essential., Methods: This study was conducted for a period of 1 y. Water samples were collected every month from 4 locations where water pools formed after rains, on the campus of a university in Chennai, South India. The water samples were monitored for Vibrio species, and characterized isolates were screened for enterotoxigenicity., Results: Thirty isolates of Vibrio cholerae belonging to a variety of serogroups and 11 strains of Vibrio species other than cholerae were isolated from the rainwater pools. On polymerase chain reaction (PCR) screening, while all the strains were positive for the ompW gene, none tested positive for the ctxA gene., Conclusions: Though all the environmental isolates of V. cholerae were non-epidemic, 4 isolates demonstrated enterotoxigenicity by rabbit ileal loop method and antibiotic resistance to drugs. This is of concern and underscores the importance of screening environmental specimens and improving civic infrastructure to prevent prolonged water-logging in developing countries.

This study was aimed to find out the host-parasite relationship between Paragonimus heterotremus isolated as metacercariae from mountain crabs, Indochinamon manipurensis, in Manipur, India and laboratory animals such as puppies, albino rats, Swiss mice, guinea pigs, and rabbits, as experimental animals. The animals were fed with the metacercariae. Infected animals were sacrificed 35 to 430 days after feeding to recover worms, which were used to determine the developmental stages. Adult worms (n = 14) were recovered from 3 puppies > or = 70 days after feeding and immature worms (n = 25) were recovered from 2 other puppies 35 or 43 days after infection. The infection rate in puppies was 100%. Juvenile worms were recovered from 3 of 13 rats: 1 of 11 rats whose viscera and cavities were examined and both of two rats whose muscles were examined. Rats were not a suitable animal model for pulmonary infection with P. heterotremus. Mice, guinea pigs, and rabbits were also found to be insusceptible to pulmonary infection with P. heterotremus.

A monoclonal antibody (MAb) specific for the bluetongue virus (BTV) group specific antigen (VP7) was characterized for its reactivity with purified virus and recombinant BTV VP7 (rVP7) protein and its suitability for use in the sandwich ELISA. The MAb, designated as 5B5 was specific to VP7 and belongs to IgG2a subclass and was selected for the development of the sELISA in this study. The MAb had a titer of 1:25 with BTV and 1:2 with the rVP7 protein. The sELISA is based on capturing of BTV antigen with VP7 specific MAb followed by detection using BTV polyclonal antiserum raised in rabbits. The assay was evaluated with six cell culture adapted serotypes of BTV that have been isolated from India, 1, 2, 15, 17, 18 and 23. The assay could detect BTV antigen as early as day 8 in blood. It was also successfully applied for the detection of BTV group specific antigen in clinical samples of blood, washed RBCs, buffy coat and plasma. A total of 102 field samples from animals, suspected of being infected with BTV, were tested and 29.42% were positive. The blood samples were also amplified in cell culture which improved the sensitivity of the assay. Results confirmed that the sELISA is rapid and specific.

The C-terminal 19 and 42 kDa fragments of Plasmodium falciparum merozoite surface protein 1 (MSP-1) have shown to be protective in animals against lethal parasite challenge. The MSP-1(19) being highly conserved may lack sufficient number of T-cell epitopes in order to elicit a broader response in genetically diverse populations. The inclusion of additional epitopes from the N-terminal MSP-1(42) has shown to enhance the protective efficacy of MSP-1(19) vaccine. In an attempt to examine the strain specific immunogenicity to MSP-1, we have cloned and expressed three diverse allelic variants of MSP-1(42) from Indian P. falciparum isolates in bacteria. Among three alleles, one was extremely rare and not been found before. These purified and refolded recombinant products were recognized by conformation specific monoclonal antibodies and hyper-immune sera. Immunization of mice and rabbits with the purified proteins generated high titer biologically active polyclonal antibodies supporting further development of this vaccine candidate antigen., ((c) 2010 Elsevier Ltd. All rights reserved.)

A polyclonal antibody-based sandwich ELISA (s-ELISA) was developed for the detection of bluetongue viruses (BTV) in cell culture lysates and blood samples of sheep infected experimentally. Rabbit antiserum to purified BTV particles and guineapig antiserum to core particles were used as capture antibody and detection antibody respectively. The assay has detected several of the BTV serotypes isolated in India so far. Other common viruses of small ruminants did not cross-react in the assay. The analytical sensitivity of the assay was estimated to be between 10(2.4) and 10(2.6)TCID(50)/ml with different serotypes of BTV. The sensitivity was compared with that of the reverse transcription polymerase chain reaction (RT-PCR) and the latter was found to be at least 100 times more sensitive. In the infected sheep, BTV antigen(s) was detected in blood as early as on 5-day post-infection (dpi) till 35 dpi. The assay may be useful for testing large number of samples in a very short time.

The YopM is an essential virulence effector produced by the bubonic plague bacterium. Yersinia pestis specific PCR gene was developed using 780 bp fragment of yopM gene. The PCR product was further cloned (in pUC57) an subcloned (pQE32 expression vector) and transformed in SG13009 E. coli host cells. The IPTG-induced recombinant protein was expressed at approximately 32 kDa region by SDS-PAGE. The recombinant protein was with 80% purity and 3mg/mL of concentration. Polyclonal and monoclonal antibodies (MAb) were generated. A total number of nine specific monoclonal antibodies obtained reacted at 43 kDa native protein of Y. pestis. Both the PCR-based assay and immunoassays were evaluated on Indian Y. pestis strains. Isolates recovered from outbreak region were positive, whereas isolates recovered from the surveillance region were negative (except one) by yopM gene PCR- and MAb-based dot-ELISA. The PCR- and ELISA-based systems developed in the present study might be utilized for detection or strain typing of Y. pestis alone or in conjunction with virulence markers such as F1 (fraction 1) and Pla (plasminogen activator).

Background: The human polyoma virus, also known as the JC virus (JCV), replicates predominantly in the oligodendrocytes, the myelin producing cells in the central nervous system and results in the fatal demyelinating disease progressive multifocal leukoencephalopathy (PML) especially in immunosuppressed patients with AIDS. Several investigators have also documented the presence of the viral genome and early and late antigens in a variety of brain tumors particularly in medulloblastomas, gliomas and ependymomas. Reports also indicate the presence of JCV in patients with colon cancer. The T antigen of JCV has been postulated to have oncogenic potential as substantiated by animal experiments. Although JCV infects 80% of the population, there are scant epidemiological studies regarding JCV from India. There are also reports of the low prevalence of PML in patients with AIDS from India and Africa., Aim: This study was undertaken to investigate if Indian children with medulloblastomas also show evidence of JCV., Methods: Twenty-two consecutive cases of medulloblastomas were investigated for the presence of T antigen and agnoprotein of JCV in biopsy specimens by immunohistochemistry. Antibodies to the agnoprotein antigen raised in rabbits and a monoclonal antibody against SV40 T antigen raised in mice that cross-reacts with JCV T antigen were used., Results: Out of 22 patients, 4 had desmoplastic tumors while the rest had classical tumors. All children were below the age of 10. Results indicate that while PML tissues showed consistent immunostaining both with antibody to T antigen and agnoprotein antibody, none of the tumors showed any positive staining for JC viral antigens., Conclusion: JCV antigens could not be detected by immunohistochemistry in the tumor tissues of Indian children with medulloblastomas.

The recent type A foot and mouth disease virus field isolates recovered in India are shown to be antigenically quite divergent from the in-use vaccine strain (IND 17/82), warranting the selection of a suitable vaccine strain which can cover this diversity in antigenic spectrum. In earlier studies employing neutralization test with anti-146S rabbit sera raised against eight candidate vaccine strains, IND 81/00 and IND 40/00 belonging to genotype VII were found to offer the best antigenic coverage. In order to assess the credibility of IND 81/00 and IND 40/00 as vaccine strains, 17 recent isolates received during 2005-2006 and representative isolates from older genotypes were subjected to two-dimensional micro-neutralization assay using bovine convalescent serum (against IND 81/00 and IND 40/00) and bovine vaccinate serum (against IND 40/00). From the results it is evident that both the isolates IND 81/00 (antigenic relationship 'r-value' >0.40 with 86% of isolates) and IND 40/00 ('r-value' >0.40 with 78% of isolates) show nearly equal antigenic relatedness with the recent field viruses and hence both of these are effective vaccine candidates in present context. Though very limited in its extent, these useful data obtained with antisera raised in homologous host system are logical extension of the on going quest for the appropriate vaccine strain and circumvents species disparities in the immune recognition of epitopes.

Rabies a disease as old as our civilization, continues to be the most feared of all communicable diseases. Despite the availability the state-of-the-art tools which ensure near cent percent protection against rabies, India is the largest contributant to rabies mortality in the world. A multicentric study was carried out from April 2001 to September 2002 with the objective of assessing the knowledge, attitudes, beliefs and practices (KAP) about animal bites and rabies in the general community. The proforma for interviewing the general community was developed and used after field testing. The study was carried out at six selected centres across the country viz. Delhi, Hyderabad, Raipur, Jamnagar, Coonoor and Rajahmundry and was co-ordinated by National Institute of Communicable Diseases (NICD), Delhi, after thorough briefing of designated nodal officers. A total of 1129 (male: female :: 48.5: 51.5) persons in the age group of 18 to 80 years were interviewed in this study. Of these about 751% of the individuals had attended school at some level and rest were illiterates. 68.7% people had heard about rabies. In 60.7% of cases the community associates rabies with dog bite only. Knowledge about appropriate wound toilet was found to be inadequate. Only 360 (31.9%0/) people felt that washing the wound with soap and water was the best option. Application of indigenous products like chillies (11.4%), turmeric (5.6%), lime (6.8%), kerosene oil (2.3%), herbal paste (4.2%) etc was suggested along with visit to occult medicine practitioner (1.5%) as part of the bite wound management. People were not aware of number of injections needed for treatment of animal bites. Multiple reasons viz negligence and ignorance 354 (31.4%), fear of multiple painful injections 365 (32.3%), expensive treatment 169 (15%) and long course requiring daily visits to anti-rabies clinics 73 (6.5%) were cited as reasons for non-compliance of treatment. KAP study suggests that there is need to create awareness amongst the masses regarding epidemiology of the disease and merits of prompt and appropriate post exposure treatment through enhanced IEC activities.

An indirect competitive inhibition enzyme-linked immunosorbent assay was reported to detect krait venom in human specimens of forensic origin. Polyclonal anti-krait venom antibodies were characterized by indirect antibody capture assay. The calibration plot was constructed based on linear regression analysis (y = 72.85 - 12.29x, r(2) = 0.98) with concentration ranges from 0.013 to 1000 ng/well of krait venom with a limit of detection of 0.2 ng/mL in the assay system. The IC50 (inhibitory concentration at 50% displacement) value of krait venom was observed to be 70 ng. Spiking studies indicated recoveries of 95-100% and 94-100% when various concentrations of krait venom were spiked to rat tissues (skin, liver, and kidneys) and pooled human serum, respectively. Polyclonal anti-krait venom antibodies showed no cross-reactivity with cobra and viper venom when tested in the assay system. The coefficient of variation of various concentrations of working range in intra-assay (n = 6) was <5%, whereas in interassay (n = 6) it was observed to be < or 7%. Further, the method was used to quantitate krait venom in human autopsy and biopsy specimens of forensic origin. Concentration of krait venom was found to be in the range of 4-172 ng/100 mg skin or skin scrapings and 64-378 ng/mL blood or serum. The methodology may find application in forensic laboratories to assess the cause of death in the cases of krait-bite victims.

This study was conducted to understand the pathophysiological changes in experimental rabbit ileal loop model using the Vibrio cholerae strain non-O1non-O139, isolated as sole pathogen from clinically diagnosed cholera patients in Kolkata. Significant amount of haemorrhagic fluid accumulation was observed in all the test loops of rabbit model where the strain of V.cholerae was inoculated as compared to control loops. Microscopic examination of the accumulated fluid showed the presence of erythrocytes and pus cells. Histology revealed structural alteration of the villous epithelium with inflammatory cells infiltration in all the layers of the gut mucosa including the nerve plexus region. Preliminary observation with a haemagglutinin protease extracted from the non-O1 non-O139 strain, was also studied in different concentrations in the same animal model which showed similar type of macroscopic and microscopic response in the ileal loops as seen with the original strain. The results highlight that along with other pathways, inflammatory cells and the enteric neurons have an important role in the pathophysiology of diarrhoea and the isolated protease may be the probable virulence factor in initiating the disease process in this non-O1non-O139 strain induced cholera.

Background & Objective: Early diagnosis is the key to the treatment of leptospirosis. For development of rapid diagnostic kits, a thorough knowledge about the nature of the proteins expressed by the pathogen during infection is necessary. The present study was undertaken to understand the nature of immunoreactive proteins from commonly circulating serogroups of Leptospira in the endemic Andaman and Nicobar Islands, India., Methods: Proteins were extracted from six strains of Leptospira representing five different serogroups following four different preparation methods, viz., whole cell lysis by sonication, detergent solubilization, outer and inner membrane isolations, and were subsequently characterized on sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Immunoblots were made from the sonicated proteins using hyperimmune rabbit antisera, homologous and heterologous patient sera separately., Results: The 67, 65, 45, 43, 35, 32 and 18 kDa major proteins in the whole cell lysate were common among all the five serogroups of Leptospira. The 67, 41, 35, 32, 28 and 22 kDa were the major outer membrane proteins, while 94, 32, 25 and 18 kDa protein were in inner membrane. Immunoblots with hyperimmune rabbit antisera detected 67, 65, 60, 45, 43, 41 and 32 kDa common proteins from the whole cell lysates of all strains while homologous and heterologous patient sera detected 32 kDa as the major immunoreactive protein in all pathogenic serogroups. This protein reacted against specific LipL32 antisera indicating that this protein was LipL32., Interpretation & Conclusion: The circulating serogroups of Leptospira have common nature of expression of proteins during human infection. Among several immunoreactive proteins, three (67, 45 and 32 kDa) were recognized as major antigens by both rabbit hyperimmune sera and patients sera while the 32 kDa protein was recognized as the major immunoreactive protein by homologous and heterologous patient sera. These conserved immunoreactive proteins could be utilized in developing indigenous diagnostic tests for leptospirosis.

Monoclonal antibodies (MAbs) were generated against the recombinant plasminogen activator (Pla) protein of Yersinia pestis. These MAbs detected Pla in all the 18 isolates of Y. pestis obtained from the sputum of pneumonic plague patients and from the liver and spleen of rodents from plague-affected areas of India during 1994-1995 as well as in seven of the eight isolates obtained from rodents in the surveillance regions of Hosur and Palmner in India during 1998 by simple dot-ELISA. In immunoblotting, the MAbs reacted with the Pla antigen only in Y. pestis isolates at 37 and 35kDa region. These monoclonal antibodies, being strictly specific, can be used for detecting Y. pestis isolates that are Fraction 1 antigen-negative. Also, the radiolabelled pla fragment hybridized specifically to the representative DNA samples of Y. pestis isolates.

A representative number of 217 camels (Camelus dromedarius) from different areas of western Rajasthan State, India, were examined from July 2002 to May 2003 for Trypanosoma evansi infection. The tests used were parasitological (wet blood film, WBF; stained thin blood smear, TBS), immunodiagnostic (double antibody sandwich enzyme linked immunosorbent assay for antigen detection, Ag-ELISA), and DNA amplification by polymerase chain reaction (PCR). These techniques were compared and the best efficiency was found for the last named (PCR). A prevalence of T. evansi infection was detected in 17.05, 9.67, 4.60 and 4.14% by PCR, Ag-ELISA, TBS and WBF with a sensitivity of 100, 56.75, 27.02 and 24.32%, respectively. PCR revealed a specific 227bp band in positive samples. The intensity of PCR bands was variable in different test samples depending upon the level of infection in the test samples. The history of intermittent fever, emaciation, oedema, poor body condition significantly correlated with positive serological status in ELISA as well as trypanosome DNA detection by PCR.