Characterization of a heterodimeric GH2 β-galactosidase from Lactobacillus sakei Lb790 and formation of prebiotic galacto-oligosaccharides

The lacLM genes from Lactobacillus sakei Lb790, encoding a heterodimeric β-galactosidase that belongs to glycoside hydrolase family GH2, were cloned and heterologously expressed in Escherichia coli . Subsequently, the recombinant β-galactosidase LacLM was purified to apparent homogeneity and characterized. The enzyme is a β-galactosidase with narrow substrate specificity because o-nitrophenyl-β-D-galactopyranoside (oNPG) was efficiently hydrolyzed, whereas various structurally related oNP analogues were not. The K(m) and k(cat) values for oNPG and lactose were 0.6 mM and 180 s(-1) and 20 mM and 43 s(-1), respectively. The enzyme is inhibited competitively by its two end-products D-galactose and D-glucose (K(i) values of 180 and 475 mM, respectively). As judged by the ratio of the inhibition constant to the Michaelis constant, K(i)/K(m), this inhibition is only very moderate and much less pronounced than for other microbial β-galactosidases. β-Galactosidase from L. sakei possesses high transgalactosylation activity and was used for the synthesis of galacto-oligosaccharides (GalOS), employing lactose at a concentration of 215 g/L. The maximum GalOS yield was 41% (w/w) of total sugars at 77% lactose conversion and contained mainly non-lactose disaccharides, trisaccharides, and tetrasaccharides with approximately 38, 57, and 5% of total GalOS formed, respectively. The enzyme showed a strong preference for the formation of β-(1→6)-linked transgalactosylation products, whereas β-(1→3)-linked compounds were formed to a lesser extent and β-(1→4)-linked reaction products could not be detected.     

Accumulative effects of humic acid and multinutrient foliar fertilizers on the vegetative and reproductive attributes of citrus (Citrus reticulata cv. kinnow mandarin)

Abstract

Field trials were performed to investigate the effects of humic acid (HA) and multinutrient foliar fertilizer “Micro Power” (MP) coupled with farmer’s practices ( FP ), addressed in single and/or split dose frames at different plant phenological stages on various vegetative, reproductive, and physiological attributes of citrus trees (Citrus reticulata cv. kinnow mandarin). The results exhibited a profound response of treatments on various growth parameters (32.5% increase in plant height, 22.2% increase in fruit set branch^?1, 5.25% decrease in fruit drop percentage, 89.81% increase in fruit yield (kg), etc.) of citrus trees when compared to the control (FP). Likewise, a significant positive response was observed regarding various plant physiological parameters (leaf nutrients, total chlorophyll content, etc.) and physicochemical characteristics (ascorbic acid, total sugars, etc.) of citrus fruits. This study confirmed the reproducibility of HA and MP applications to improve the yield/quality of citrus and can lead to an organically sustainable citriculture.     

Effect of short-term supply of farmyard manure on maize growth and soil parameters in pot culture

Farmyard manure (FYM) improves various soil parameters and to a large extent, the availability of water and nutrient to crops when it is applied to the soil. This study aims to further investigate the short-term effects of different levels of FYM on maize plants and soil parameters. Maize plants grown in pot culture were treated with no FYM (control), recommended NPK (inorganic fertilizers), and FYM at 2, 4, 6, 8, and 10 t ha^?1 along with recommended NPK, and the cultures were analyzed 8 weeks after germination. Soil bulk density and soil pH decreased with the increasing levels of FYM, whereas soil porosity, soil organic matter (SOM), soil water content, plant height, root and shoot yield, and NPK uptake of maize were increased compared with the control or recommended NPK, respectively. The present results indicate that short-term application of higher FYM levels improves soil properties. Furthermore, the application of FYM at only higher rates significantly increases the nutrient uptake of maize plants due to improved soil properties. The supply of different amounts of nutrients increases biomass and nutrient uptake in plants.     

Glyphosate-tolerant cotton in Australia: successes and failures

ABSTRACT

Cotton is an important cash crop grown on 2.5% of the world’s arable land in over 100 countries, and has a 31% share of the world’s fibre market. In Australia, cotton is also a leading crop and contributes around AUD $3 billion to the total agricultural production. Weeds are a major biotic constraint resulting in yield losses of up to 90% and revenue losses of around AUD $100 billion globally and $4 billion to Australian agriculture. Genetically-modified (GM) crops have refashioned the weed management with more dependency on glyphosate. Such overreliance has led to the evolution of 43 glyphosate-resistant (GR) weed populations globally, with 16 species reported from Australia. Such GR weeds along with volunteer glyphosate-tolerant (GT) plants are now decreasing the value of the GM crops and forcing growers to spend more time and effort, and investment in their management. Weed management strategies need to be diversified and integrated with non-chemical methods and alternative herbicides not only to achieve efficient control, but to reduce the rate of evolution of GR weeds. In future, research is needed to improve integrated weed management through development and use of competitive and multiple herbicide-tolerant (HT) crops, organic herbicides, bio-herbicides, RNAi technology and robotics.     

Aflatoxins and ochratoxin A contamination in rice,corn and corn products from Punjab,Pakistan

In present study, a total of 275 retail cereal samples including rice, corn and corn products was analyzed for the presence of aflatoxins and ochratoxin A (OTA) using HPLC equipped with a fluorescence detector. The data has shown that 38 out of 68 samples of rice, 37/105 of corn and 43/102 samples of corn products have been found contaminated with AFs, average level of AFB₁ and total AFs, 8.23 and 19.54, 7.90 and 12.08, and 5.47 and 7.85 μg/kg, respectively. Furthermore, 34, 28 and 26 numbers of rice, corn and corn product samples were found to be contaminated with OTA with an average level of 12.94, 5.29 and 3.69 μg/kg, respectively. The samples of rice, corn and corn products found above the permissible European Union (EU) limit for AFB₁ and total AFs were 18, 13 and 14 and 28, 14 and 20%, respectively; however, the samples of rice, corn and corn products above the EU limit were 40, 14 and 15%, respectively.     

Prospects of raising Sahiwal cow calves for veal production under tropical environment

The objective of present study was to evaluate the growth potential of Sahiwal calves for veal production on whole milk or whole milk and milk replacer combined in a ratio of 50:50 (MMR). For this purpose, 48 Sahiwal calves (both male and female) were assigned to four dietary treatments having 12 animals/treatment. Calves in the treatments A and B were offered whole milk at 15 or 20 % of their body weight (BW), respectively, up to day 84 adjusted on weekly basis. The calves in treatments C and D received the same amount of milk as in treatments A and B until day 21, respectively, after which 50 % of the milk offered was replaced with a blend of chickpea (Cicer arietinum) flour and vegetable (corn) oil mixed in water (MR) until day 84. The constituted MR had 3.1, 2.8, and 14.3 %, CP, EE, and DM, respectively. The growth and intake data were analyzed using repeated measures analysis, with MIXED Procedures of SAS in a 2?×?2 factorial design. The two factors were feeding level and feeding source. Calves offered whole milk grew faster (P?<?0.05) and had greater weaning weights (P?<?0.05) than those offered MMR (606.4?±?18.1 vs 331.3?±?18.1 g/day and 70.4?±?1.5 vs 47.8?±?1.5 kg, respectively). Greatest daily BW gain (656?±?26 g/day) and weaning weight (74.6?±?2.1 kg) were observed in calves given treatment B while the lowest production cost/kg of BW gain (≈US$3.6?±?0.2) was observed in calves given treatment A. The daily BW gain of calves fed milk ad libitum was 716?±?40 and 836?±?40 g/day, during 5–8 and 9–12 weeks, respectively. The number of days calves exhibited scours was higher in calves offered MMR than those offered whole milk. Replacement of 50 % milk with a blend of chickpea flour and vegetable oil, as an alternative to milk replacer, did not support growth equivalent to whole milk and was not effective in reducing feeding cost during the weaning period. Sahiwal calves may have promise for being raised for veal production under tropical environments.     

Exploitation of Marine-Derived Robust Biological Molecules to Manage Inflammatory Bowel Disease

Naturally occurring biological entities with extractable and tunable structural and functional characteristics, along with therapeutic attributes, are of supreme interest for strengthening the twenty-first-century biomedical settings. Irrespective of ongoing technological and clinical advancement, traditional medicinal practices to address and manage inflammatory bowel disease (IBD) are inefficient and the effect of the administered therapeutic cues is limited. The reasonable immune response or invasion should also be circumvented for successful clinical translation of engineered cues as highly efficient and robust bioactive entities. In this context, research is underway worldwide, and researchers have redirected or regained their interests in valorizing the naturally occurring biological entities/resources, for example, algal biome so-called “treasure of untouched or underexploited sources”. Algal biome from the marine environment is an immense source of excellence that has also been demonstrated as a source of bioactive compounds with unique chemical, structural, and functional features. Moreover, the molecular modeling and synthesis of new drugs based on marine-derived therapeutic and biological cues can show greater efficacy and specificity for the therapeutics. Herein, an effort has been made to cover the existing literature gap on the exploitation of naturally occurring biological entities/resources to address and efficiently manage IBD. Following a brief background study, a focus was given to design characteristics, performance evaluation of engineered cues, and point-of-care IBD therapeutics of diverse bioactive compounds from the algal biome. Noteworthy potentialities of marine-derived biologically active compounds have also been spotlighted to underlying the impact role of bio-active elements with the related pathways. The current review is also focused on the applied standpoint and clinical translation of marine-derived bioactive compounds. Furthermore, a detailed overview of clinical applications and future perspectives are also given in this review.     

Improvement of wheat (Triticum aestivum) drought tolerance by seed priming with silicon

Silicon has the potential to improve drought tolerance in crops. Seeds primed with silicon were used in the present study to explore its potential benefit to withstand water stress. Seeds of two wheat varieties, NARC-2009 and Chakwal-50, were sown in pots after priming with distilled water and different concentrations (0.5%, 1.0% and 1.5%) of silicon sources (silicic acid, sodium silicate and silica gel) at PMAS, Arid Agriculture University, Rawalpindi. Maximum silicon uptake at three-leaf stage (0.028 µg g^?1 dry weight (DW)), anthesis (0.072 µg g^?1 DW) and maturity (0.103 µg g^?1 DW) was recorded for silica gel. Silicon uptake increased significantly in response to increase in Si concentration from 0.5% to 1.5%. Leaf membrane stability index, epicuticular wax, relative water content and proline remained maximum – 78.90%, 2.6 mg g^?1 DW, 83.88% and 54.90 µg g^?1 – for silica gel treatments compared with others. Silica gel with 1.5% silicon concentration resulted in maximum spike length (14.3 cm), biological yield (7.63 g pot^?1), hundred-grain weight (3.97 g pot^?1) and grain yield (2.46 g pot^?1). Based on the study outcomes, it is concluded that silica gel might be a good priming option with 1.5% silicon concentration to establish plant under drought stress.     

The digestive fate of Escherichia coli glutamate dehydrogenase deoxyribonucleic acid from transgenic corn in diets fed to weanling pigs

Corn containing genetically engineered plasmid DNA encoding an Escherichia coli glutamate dehydrogenase (gdhA) was fed to 19-d-old weanling swine to trace the digestive fate of the transgenic DNA. Eight pens of 8 pigs were fed a commercial (nongdhA) starter for 2 wk. One pig was randomly selected from each pen for 0-h control samples. The remaining 56 pigs were transitioned onto a corn-soybean meal diet and fed a diet containing 58% gdhA corn for approximately 1 wk; immediately thereafter, liver, 10th rib muscle, white blood cells, and plasma from the hepatic portal vein and ingesta from the stomach, distal ileum, and large intestine were collected. The DNA was extracted and the concentration determined via spectrophotometry. Polymerase chain reaction and gel electrophoresis were performed with primers designed to amplify 490 bp that included the plasmid's ligation site between the maize ubiquitin and the gdhA genes. The gdhA corn-derived DNA and diet served as positive assay controls, and conventional corn DNA and distilled water acted as negative assay controls. Detection limits were 0.99 fg of target DNA confounded with 500 ng of conventional corn DNA per each 20 &L reaction. Transgenic DNA was detected in 71.43% of the stomach and 1.79% of the ileal ingesta samples from treatment animals but was not detected in the large intestine, white blood cells, plasma, liver, or muscle samples. Transgenic DNA was not detected in any sample from 0-h control animals. Stomach and ileal ingesta samples were further analyzed using real-time PCR. With an estimated limit of detection of 1.049 ag/microL, 89.29% of the stomach ingesta samples were positive (average 1.56 fg target DNA). The proportion of transgenic DNA to total DNA differed between diet and stomach ingesta samples (P < 0.001). Despite the greater sensitivity of real-time PCR, target DNA was detected in only 1.79% of ileal ingesta. These data suggest that the gdhA transgene began degradation in the stomach and was nondetectable in the large intestine.