Risk factor analysis associated with Theileria equi infected equines in semi-arid and sub-humid ecological enzootic zones of India.

Equine piroplasmosis is a haemoprtozoan disease of equines and enzootic in tropical and subtropical countries. A cross-sectional study on sero-prevalence of Theileria equi, the causative agent of EP, was performed in semi-arid and sub-humid ecological endemic zones of India including Rajasthan, Haryana and Gujarat states, in order to evaluate the enzootic status/level of exposure to equine population due to this infection and addressed associated risk factors. Serum samples were collected from a total of 1021 equids that comprised of 792 horses, 168 donkeys and 61 mules and evaluated for T. equi specific antibodies in equine merozoite antigen-2 (EMA-2) based on indirect ELISA. The state with high sero-prevalence rate was Rajasthan (71.40%), followed by Haryana (60.39%) and Gujarat (48.92%). Overall T. equi sero-positivity in equines was 64.44%. Species-wise T. equi sero-prevalence was 66.29%, 51.19% and 91.80% in horses, donkeys and mules respectively. The association and risk factor among age, gender and species in relation with T. equi infection was statistically analyzed at 95% level of significance (p<0.05). A very high T. equi sero-positivity was recorded in 0-1year age group of equines (60%), indicating that this naïve age group contacts with T. equi infected ticks and remains infected throughout its lifetime. The sero-prevalence rate was significantly associated with the species of animal (p<0.05). The risk factor analysis kept mules at higher risk (Odd's ratio; 5.696; 95% confidence interval: 2.25-14.38) of getting infection as compared to horses and donkeys. This study has demonstrated high enzootic nature of T. equi infection in semi-arid and sub-humid ecological zones of India. Mules, in comparison to horses and donkeys were found at higher risk of getting T. equi infection, indicating that disease prevalence is associated with species of the infected host.

Fermented pearl millet (Pennisetum glaucum) with in vitro DNA damage protection activity, bioactive compounds and antioxidant potential.

In the present study, pearl millet cultivar PUSA-415 was fermented by solid state fermentation (SSF) process using Aspergillus sojae (MTCC-8779) as starter culture. The fermentation was carried out for the period of ten days. The effect of SSF on phenolic content, condensed tannin content, antioxidant potential and DNA damage protection of pearl millet during different fermentation period was determined. Results showed that SSF and thermal processing significantly affect the bioactive profile and antioxidant potential of bio-transformed pearl millet. Extracts prepared from 6th days fermented pearl millet flour exhibited the highest TPC, antioxidant potential and DNA damage protection activity. Qualitative and quantitative analysis of bioactive compounds were done by HPLC. During SSF, production of enzymes (α-amylase, ß-glucosidase and xylanase) as well as specific bioactive compounds (ascorbic acid, gallic acid and p-Coumaric acid) was significantly increased. Thus, bio-transformed Aspergillus sojae fermented pearl millet could be used in preparation of functional foods and novel nutraceuticals in health promotions. Chapatti was formulated from unfermented as well as fermented flour and the effect of thermal processing on bioactive compounds and antioxidant potential was studied. Thermal processing resulted in decrease in TPC of both, AFM and UFM by 4.75-16.27% and increase in CTC by 38.52-67.41%.

Bioactive profile, free-radical scavenging potential, DNA damage protection activity, and mycochemicals in Aspergillus awamori (MTCC 548) extracts: a novel report on filamentous fungi.

Biomass of Aspergillus awamori was investigated for mycochemicals, total phenolic compounds (TPC), condensed tannin content (CTC), free-radical scavenging potential (FRSP), and DNA damage protection activity. FRSP was determined using DPPH, ABTS, FRAP (Ferric reducing antioxidant power), metal chelating activity, and cupric reducing antioxidant capacity) assays. Water (Aq), aqueous ethanol 50% (AqE), and methanol were used as extraction phase at 44.5 °C for 23.8 min. AqE shows the presence of maximum mycochemicals (coumarins, glucose, saponins, flavonoids, and tannin). Further quantitative analysis shows maximum TPC (23.17 mg GAE/g dwb) in AqE and CTC (.89 mg CE/g dwb) in ME. Qualitative and quantitative analysis for identification of specific bioactive compound in AqE was carried out using HPLC. HPLC analysis confirmed the presence of bioactive compounds: p'-Coumaric acid (5.96 mg/g dwb), cinnamic acid (4.31 mg/g dwb), gallic acid (2.27 mg/g dwb), and ascorbic acid (.98 mg/g dwb). All the extracts show significant DNA damage protection activity; however, AqE showed the maximum activity. Pearson correlations were also calculated to find the relationships between bioactive compounds and antioxidant potential.

Improved production of tannase by Klebsiella pneumoniae using Indian gooseberry leaves under submerged fermentation using Taguchi approach.

Tannase (tannin acyl hydrolase E.C 3.1.1.20) is an inducible, largely extracellular enzyme that causes the hydrolysis of ester and depside bonds present in various substrates. Large scale industrial application of this enzyme is very limited owing to its high production costs. In the present study, cost effective production of tannase by Klebsiella pneumoniae KP715242 was studied under submerged fermentation using different tannin rich agro-residues like Indian gooseberry leaves (Phyllanthus emblica), Black plum leaves (Syzygium cumini), Eucalyptus leaves (Eucalyptus glogus) and Babul leaves (Acacia nilotica). Among all agro-residues, Indian gooseberry leaves were found to be the best substrate for tannase production under submerged fermentation. Sequential optimization approach using Taguchi orthogonal array screening and response surface methodology was adopted to optimize the fermentation variables in order to enhance the enzyme production. Eleven medium components were screened primarily by Taguchi orthogonal array design to identify the most contributing factors towards the enzyme production. The four most significant contributing variables affecting tannase production were found to be pH (23.62 %), tannin extract (20.70 %), temperature (20.33 %) and incubation time (14.99 %). These factors were further optimized with central composite design using response surface methodology. Maximum tannase production was observed at 5.52 pH, 39.72 °C temperature, 91.82 h of incubation time and 2.17 % tannin content. The enzyme activity was enhanced by 1.26 fold under these optimized conditions. The present study emphasizes the use of agro-residues as a potential substrate with an aim to lower down the input costs for tannase production so that the enzyme could be used proficiently for commercial purposes.

Optimization of tannase production by a novel Klebsiella pneumoniae KP715242 using central composite design.

A novel tannase producing bacterial strain was isolated from rhizospheric soil of Acacia species and identified as Klebsiella pneumoniae KP715242. A 3.25-fold increase in tannase production was achieved upon optimization with central composite design using response surface methodology. Four variables namely pH, temperature, incubation period, and agitation speed were used to optimize significant correlation between the effects of these variables on tannase production. A second-order polynomial was fitted to data and validated by ANOVA. The results showed a complex relationship between variables and response given that all factors were significant and could explain 99.6% of the total variation. The maximum production was obtained at 5.2 pH, 34.97 °C temperature, 103.34 rpm agitation speed and 91.34 h of incubation time. The experimental values were in good agreement with the predicted ones and the models were highly significant with a correlation coefficient (R2) of 0.99 and a highly significant F-value of 319.37.

Stress influenced increase in phenolic content and radical scavenging capacity of Rhodotorula glutinis CCY 20-2-26.

Rhodotorula glutinis CCY 20-2-26 when grown under controlled stress of either NaCl (1-5 %) or H2O2 (1-5 mM) on basal media exhibited a twofold increase in its total phenolic contents. The radical scavenging capacities (RSCs) as determined by ABTS test were found to be highest in 4 mM H2O2 (1.44 mM TEAC mg-1) and 4 % NaCl (1.13 mM TEAC mg-1) as compared to control samples (0.41 mM TEAC mg-1). Similarly, the RSCs as determined by DPPH test were also highest in 4 % NaCl (1.83 mM TEAC mg-1) and 4 mM H2O2 (1.78 mM TEAC mg-1) compared to control (0.48 TEAC mg-1). The relative RSCs from EPR spin-trapping assay for H2O2-stressed cultures were highest in 1 mM H2O2 (56.1 µM TEAC g-1) whereas in NaCl-stressed cultures it was highest in 5 % NaCl (44.6 µM TEAC g-1) as compared to control (30.9 µM TEAC g-1). Five phenolic compounds (gallic acid, benzoic acid, catechin, caffeic acid and ferulic acid) were detected for the first time in R. glutinis CCY 20-2-26.

Studies on leaf spot disease of Withania somnifera and its impact on secondary metabolites.

During an investigation of the disease profile of Withania somnifera, it was observed that leaf spot is the most prevalent disease. Repeated isolations from infected leaf tissues and pathogenicity tests showed the association of fungal pathogen identified as Alternaria alternata (Fr.) Keissler. Scanning electron microscopy showed various histological changes in the leaf tissues of infected plants. A decrease in total content of reducing sugars (20%) and chlorophyll (26.5%) was observed in diseased leaves whereas an increase was noticed in proline (25%), free amino acids (3%) and proteins (74.3%). High performance thin layer chromatography (HPTLC) analysis of secondary metabolites viz. withanolides, withaferin-A and total alkaloids of the diseased leaves vis-à-vis control revealed reduction in withaferin-A and withanolides contents by 15.4% and 76.3% respectively, in contrast to an increase in total alkaloids by 49.3%, information hitherto unreported in W. somnifera.