ABSTRACT Aspergillus fumigatus is a respiratory pathogen, which has the potential to cause brooder pneumonia in poultry chicks and aspergillosis in birds and farmers. Frequent and irrational use of antifungals has led to the development of resistance in the fungal species. The present study aimed to evaluate the antifungal activity of essential oils (EOs) from four different spices against multi-drug resistant (MDR) A. fumigatus isolated from 40 poultry feed samples. Twenty fungal isolates were identified to be A. fumigatus based on the macroscopic and microscopic characteristics, which were confirmed by targeting and amplifying the species-specific rodA gene (313 bp). Five (5/20) isolates were found to be MDR by the disk diffusion method. EOs were tested as an alternative strategy for 5 MDR isolates of A. fumigatus. Clove EO provided better activity by 6 log10 reduction for 30-90 min exposure as compared to other EOs, therefore, shortlisted for well-diffusion and minimum inhibitory concentration (MIC) method. Clove EO provided promising results as compared to fluconazole, amphotericin B, ketoconazole, nystatin and voriconazole as its mean ZOI (39.6 ± 6.34) were better than all other commercial antifungals (p< 0.001). Mean MIC was reported to be 0.195 ± 0.11 µl/ml. Safety testing of the EO produced 74-82% cell viability for the range of 0.390 - 0.097 µl/ml as compared to DMSO. Findings of the present study highlight that clove EO can be source of a good therapeutic agent for infections of MDR isolates of A. fumigatus.

RESUMO Aspergillus fumigatus é um patógeno respiratório, que tem o potencial de causar pneumonia incubadora em pintos e aspergilose em aves e criadores. O uso frequente e irracional de antifúngicos tem levado ao desenvolvimento de resistência nas espécies fúngicas. O presente estudo teve como objetivo avaliar a atividade antifúngica de óleos essenciais (OEs) de quatro diferentes especiarias contra A. fumigatus multirresistente (MDR) isolado de 40 amostras de ração de aves. Vinte isolados fúngicos foram identificados como A. fumigatus com base nas características macroscópicas e microscópicas, que foram confirmadas pelo direcionamento e amplificação do gene rodA espécie-específico (313 pb). Cinco (5/20) isolados foram considerados MDR pelo método de difusão em disco. OEs foram testados como uma estratégia alternativa para 5 isolados MDR de A. fumigatus. Clove EO forneceu melhor atividade por redução de 6 log10 para exposição de 30-90 min em comparação com outros OEs, portanto, pré-selecionado para difusão e método de concentração inibitória mínima (MIC). Clove EO forneceu resultados promissores em comparação com fluconazol, anfotericina B, cetoconazol, nistatina e voriconazol, com ZOI médio (39,6 ± 6,34) melhor do que todos os outros antifúngicos comerciais (p < 0,001). A MIC média foi de 0,195 ± 0,11 µl/ml. O teste de segurança do EO produziu 74-82% de viabilidade celular para a faixa de 0,390 - 0,097 µl/ml em comparação com DMSO. Os resultados do presente estudo destacam que o EO de cravo pode ser fonte de um bom agente terapêutico para infecções de isolados multirresistentes de A. fumigatus.

Abstract The present study conducted to assess nutritional, safety parameters (aflatoxin M1/AFM1 and heavy metals; copper, cadmium and zinc) and fatty-acid composition of tea-whiteners (TWs). Accordingly, TW’s samples composition (moisture, ash, protein, fat and nitrogen free extract) determined using AOAC methods, minerals including Na, K and Ca heavy metals including Cu, Cd and Zn determined via atomic absorption spectrophotometer. The AFM1 measured using thin layer chromatography (TLC) whereas, fatty acids and trans-fat contents were determined using gas chromatography. ANOVA and independent sample t test applied to assess the significant difference. Results indicated that protein and fat contents (%) of liquid TWs were 2.7-5.4 and 4.28-6.48, respectively. Powdered TWs protein and fat (%) were 13.4-17, and 23.06-29.03, respectively. AFM1 was not found in any TW. Cu/Cd and Zn levels (ppm) of liquid TWs were 0.296-2.075, ND-1.206 and 0.009-0.026 respectively. For powdered TWs the readings (ppm) were 0.515-0.632, 0.003-0.005 and 0.010-0.013 respectively. Trans-fats (%) in liquid TWs were 2.90-7.51 and in powdered TWs were 1.29-1.83. In nutshell, TWs had traces of trans fats ranged 1.29-7.51% whereas, liquid TWs contain more trans-fats than powdered TWs. Major fatty acids among liquid and powdered TWs were palmitic acid, heptadecanoic acid, stearic acid and oleic acid.

Abstract Moringa the miracle tree is rich source of protein and its protein content are comparable with meat protein. In the current study protein was isolated from moringa flowers and utilized in preparation of bars. Physicochemical analysis like texture, color, caloric value, water activity, pH and sensory characteristics were determined to evaluate the nutritional and quality attributes of bars. Different concentration of isolated protein was used in the preparedness. Among all treatments T4 (15% protein isolate) and T3 (10% protein isolate) were best as they had the highest caloric value i.e. 498.33 and 480.37 kcal and high protein percentage 42.55 and 35.29%. T4 and T3 also were good in all textural parameters i.e. hardness 4.68 and 4.85 kg, firmness 3.88 and 3.67 kg, toughness 3.78 and 3.63 kg and work of shear 16.84 and 16.80 kg/mm. The obtained results indicated that the incorporation of moringa protein isolate in bars not only increase their aesthetic value but also increase their nutritional profile. High quality protein isolated from moringa is cheaper and easily grown with little care to provide ample supply of moringa protein to peoples which were in concern of money for provision of proteinaceous food to their families to combat malnutrition.

Kinetics of oxidation of L-lysine by permanganate ion in a perchloric acid medium was investigated to explore the order of the reaction with respect to oxidant and substrate and to study the catalytic behaviour of sodium lauryl sulphate (SLS) and polyethylene glycol (PEG). The reaction was found to be first-order with respect to the oxidant and the substrate and zero-order with respect to hydrogen ion. Changes in the sodium sulphate concentration produce a non-significant variation in the rate of the reaction. SLS and PEG were found to catalyze the reaction. Surfactant catalysis was modelled by Piszkiewicz's cooperativity model, while polymer catalysis was explained with the help of the Benesi-Hildebrand equation. The temperature dependence of the rate of the reaction was elucidated, and activation parameters were obtained. Interestingly, the reaction was found to possess positive activation entropy indicating the dissociative nature of the transition state and outer-sphere electron transfer mechanism. A mechanism of the reaction that is supported by the experimental findings was suggested.

Abstract Health safety of spirulina (Spirulina platensis) for food application was evaluated through hematology and histopathology of liver, spleen and kidney in Sprague Dawley rats fed on three isoproteic diets maintained to 10% protein for forty five days. Three groups of rats were maintained as control diet casein (R-C); diet containing crude spirulina (R-S) and diet containing purified spirulina protein (R-SPI). Hematological profile of the rats group fed on spirulina diets showed no significant increase in white blood cells (WBCs), while there was decreasing trend in the levels of neutrophils (60.46 ± 2.60%) and lymphocytes (33.53 ± 1.06%) in rats that were fed on spirulina diet (R-S) than the other tested experimental diet group. Histopathological examination revealed normal architecture of the liver with no abnormality in kidneys and spleen exhibiting safety of spirulina in both forms to mitigate protein energy malnutrition widely prevalent in developing countries.

Abstract Vitamin C is imperative component of our nutrition and used as additive in many foods owing to its high antioxidant activity. The degradation of vitamin C content in orange juice during storage was evaluated in this study. Degradation of vitamin C in orange juice during 40 days storage at 0, 05, 15, 25, and 40 °C was analyzed and evaluated. Hydroxymethylfurfural (HMF) accumulation in orange juice acts as an indicator of ascorbic acid degradation. The initial vitamin C content in orange juice was 34.26 mg/100 mL that reduced when subjected to various storage conditions such as temperature and time. The loss of vitamin C during storage was lowest at 0 °C and the losses increased with rise in temperature. The Vitamin C content after 40 days in orange juice at 0 °C were 7.19 mg/100 mL while the Vitamin C content reduced to 1.89 mg/100 mL in orange juice that were kept at 40 °C. Hence, with increase in temperature, the Vitamin C was degraded and accretion of HMF, 5 methyl furfural, and 2 methyl furfural was increased. The zero-order interaction between storage time and treatment showed significant influence on HMF.

BACKGROUND: Insects have developed resistance against Bt-transgenic plants. A multi-barrier defense system to weaken their resistance development is now necessary. One such approach is to use fusion protein genes to increase resistance in plants by introducing more Bt genes in combination. The locating the target protein at the point of insect attack will be more effective. It will not mean that the non-green parts of the plants are free of toxic proteins, but it will inflict more damage on the insects because they are at maximum activity in the green parts of plants. RESULTS: Successful cloning was achieved by the amplification of Cry2A, Cry1Ac, and a transit peptide. The appropriate polymerase chain reaction amplification and digested products confirmed that Cry1Ac and Cry2A were successfully cloned in the correct orientation. The appearance of a blue color in sections of infiltrated leaves after 72 hours confirmed the successful expression of the construct in the plant expression system. The overall transformation efficiency was calculated to be 0.7%. The amplification of Cry1Ac-Cry2A and Tp2 showed the successful integration of target genes into the genome of cotton plants. A maximum of 0.673 μg/g tissue of Cry1Ac and 0.568 μg/g tissue of Cry2A was observed in transgenic plants. We obtained 100% mortality in the target insect after 72 hours of feeding the 2nd instar larvae with transgenic plants. The appearance of a yellow color in transgenic cross sections, while absent in the control, through phase contrast microscopy indicated chloroplast localization of the target protein. CONCLUSION: Locating the target protein at the point of insect attack increases insect mortality when compared with that of other transgenic plants. The results of this study will also be of great value from a biosafety point of view.

Background: Transgenic plants inhabiting single Bt gene are prone to develop insect resistance and this resistance has been reported in case of some important yield-devastating insect larvae of commercial crops, such as cotton and rice. Therefore, it has become essential to adapt new strategies to overcome the problem of insect resistance and these new strategies should be sophisticated enough to target such resistant larvae in broad spectrum. Among these, plants may be transformed with Bt gene tagged with some fusion-protein gene that possesses lectin-binding capability to boost the binding sites for crystal protein gene within insect mid-gut in order to overcome any chances of insect tolerance against Bt toxin. Enhanced chloroplast-targeted Bt gene expression can also help in the reduction of insect resistance. Results: In the present investigation, a combined effect of both these strategies was successfully used in cotton (G. hirsutum). For this purpose, plant expression vector pKian-1 was created, after a series of cloning steps, carrying Cry1Ac gene ligated with chloroplast transit peptide towards N-terminal and Ricin B-Chain towards C-terminal, generating TP-Cry1Ac-RB construct. Conclusions: Efficacy of pKian-1 plasmid vector was confirmed by in-planta Agrobacterium-mediated leaf GUS assay in tobacco. Cotton (G. hirsutum) local variety MNH-786 was transformed with pKian-1 and the stable integration of TP-Cry1Ac-RB construct in putative transgenic plants was confirmed by PCR; while fusion-protein expression in cytosol as well as chloroplast was substantiated by Western blot analysis. Whereas, confocal microscopy of leaf-sections of transgenic plants exposed that hybrid-Bt protein was expressing inside chloroplasts.

Various cultivation parameters were optimized for the production of extra cellular protease by Brevibacterium linens DSM 20158 grown in solid state fermentation conditions using statistical approach. The cultivation variables were screened by the Plackett-Burman design and four significant variables (soybean meal, wheat bran, (NH4)2SO4 and inoculum size were further optimized via central composite design (CCD) using a response surface methodological approach. Using the optimal factors (soybean meal 12.0g, wheat bran 8.50g, (NH4)2SO4) 0.45g and inoculum size 3.50%), the rate of protease production was found to be twofold higher in the optimized medium as compared to the unoptimized reference medium.

A respiratory complex was isolated from plasma membrane of pathogenic Proteus mirabilis strain ATCC 29245. It was identified as complex II consisting of succinate:quinone oxidoreductase (EC 1.3.5.1) containing single heme b. The complex II was purified by ion-exchange chromatography and gel filtration. The molecular weight of purified complex was 116.5 kDa and it was composed of three subunits with molecular weights of 19 kDa, 29 kDa and 68.5 kDa. The complex II contained 9.5 nmoles of cytochrome b per mg protein. Heme staining indicated that the 19 kDa subunit was cytochrome b. Its reduced form showed absorptions peaks at 557.0, 524.8 and 424.4 nm. The α-band was shifted from 557.0 nm to 556.8 nm in pyridine ferrohemochrome spectrum. The succinate: quinone oxidoreductase activity was found to be high in this microorganism.