Molecular dissection of apomixis - an asexual reproductive mode - is anticipated to solve the enigma of loss of meiotic sex, and to help fixing elite agronomic traits. The Brassicaceae genus Boechera comprises of both sexual and apomictic species, permitting comparative analyses of meiotic circumvention (apomeiosis) and parthenogenesis. Whereas previous studies reported local transcriptome changes during these events, it remained unclear whether global changes associated with hybridization, polyploidy and environmental adaptation that arose during evolution of Boechera might serve as (epi)genetic regulators of early development prior apomictic initiation. To identify these signatures during vegetative stages, we compared seedling RNA-seq transcriptomes of an obligate triploid apomict and a diploid sexual, both isolated from a drought-prone habitat. Uncovered were several genes differentially expressed between sexual and apomictic seedlings, including homologs of meiotic genes ASYNAPTIC 1 (ASY1) and MULTIPOLAR SPINDLE 1 (MPS1) that were down-regulated in apomicts. An intriguing class of apomict-specific deregulated genes included several NAC transcription factors, homologs of which are known to be transcriptionally reprogrammed during abiotic stress in other plants. Deregulation of both meiotic and stress-response genes during seedling stages might possibly be important in preparation for meiotic circumvention, as similar transcriptional alteration was discernible in apomeiotic floral buds too. Furthermore, we noted that the apomict showed better tolerance to osmotic stress in vitro than the sexual, in conjunction with significant upregulation of a subset of NAC genes. In support of the current model that DNA methylation epigenetically regulates stress, ploidy, hybridization and apomixis, we noted that ASY1, MPS1 and NAC019 homologs were deregulated in Boechera seedlings upon DNA demethylation, and ASY1 in particular seems to be repressed by global DNA methylation exclusively in the apomicts. Variability in stress and transcriptional response in a diploid apomict, which is geographically distinct from the triploid apomict, pinpoints both common and independent features of apomixis evolution. Our study provides a molecular frame-work to investigate how the adaptive traits associated with the evolutionary history of apomicts co-adapted with meiotic gene deregulation at early developmental stage, in order to predate meiotic recombination, which otherwise is thought to be favorable in stress and low-fitness conditions.
Lactobacilli are probiotics with Aflatoxin (AF) detoxification ability, found in fermented products, GIT of animals and environment. Purpose of this study was to investigate the ability of broiler isolates of Lactobacillus against Aflatoxin B1 (AFB1). For this purpose, 5 isolates of Lactobacillus from broiler gut were incubated with 100 ppb AFB1 in aqueous environment and effect of different parameters (cell fractions, time, temperature, pH) on detoxification was determined by HPLC. The ameliorative effect of Lactobacillus salivarius (LS) against AFB1 was studied in broiler. The results revealed that LS (CR. 4) showed the best results (in vitro) as compared to other isolates (L. salivarius (CR. 3, CR, 4), L. agilis (CE. 2.1, CE. 3.1) and L. crispatus (CE. 28). Cell debris of CR. 4 showed significantly higher detoxification (P<0.05). Maximum amount of AFB1 was detoxified at 30°C (97%), pH 4.0 (99%) and 6 h (99.97%). In vivo study showed that AFB1 decreased weight gain (1,269 ± 0.04 gm/ bird), feed consumed (2,161 ± 0.08 gm/ bird), serum total protein (2.42 ± 0.34 gm/ dl), serum albumin (0.5 ± 0.2 2 gm/dl) and antibody titer (4.2 ± 0.83). Liver function enzymes were found (alanine transaminase (ALT): 32 ± 10.7 U/L) and aspartate transaminase (AST): 314.8 ± 27 U/L) elevated in AFB1 fed broilers. Treatment with 1% LS not only decreased the toxic effects of AFB1 (group D) but also improved the overall health of broilers due to its probiotic effects (p<0.05) as compared to control negative (group A). The detoxification ability of LS was better than commercial binder (CB) (0.2% Protmyc). It was concluded that detoxification of AFB1 by Lactobacillus was strain, temperature, pH and time dependent. LS has detoxification ability against AFB1 in vivo.
Genomic imprinting confers parent-of-origin-specific gene expression, thus non-equivalent and complementary function of parental genomes. As a consequence, genomic imprinting poses an epigenetic barrier to parthenogenesis in sexual organisms. We report aberrant imprinting in Boechera, a genus in which apomicts evolved from sexuals multiple times. Maternal activation of a MADS-box gene, a homolog of which is imprinted and paternally expressed in the sexual relative Arabidopsis, is accompanied by locus-specific DNA methylation changes in apomicts where parental imprinting seems to be relaxed.
Lactoferrin is the part of whey proteins, which does not become the part of curd and is lost in the whey. The objective of the current investigation was to develop lactoferrin-supplemented cheddar cheese. Cheddar cheese was supplemented with lactoferrin at 5, 10, 15, 20, and 0 mg/100 g concentrations (T1, T2, T3, T4, and control) and matured for 90 days. The supplementation of lactoferrin at all four concentrations did not affect the compositional attributes of cheddar cheese. In mature cheddar cheese, values of log-colony forming units in T1, T2, T3, T4, and control per mL were 7.82, 7.77, 6.51, 6.21, and 7.95, respectively. Lactoferrin contents of T1, T2, T3, T4, and control were 4.98, 9.89, 14.91, 19.88, and 0.11 mg/100 g cheese. In vitro total antioxidant capacity of mature T1, T2, T3, T4, and control were 32.47%, 47.68%, 65.37%, 78.82%, and 45.79%. High-performance liquid chromatography (HPLC) analysis revealed that concentrations of citric acid, lactic acid, acetic acid, and propionic acid in mature T4 were 2389, 16,468, 126, and 141 mg/kg. Analysis of cheese samples on gas chromatography–mass spectrometry (GC-MS) showed that fatty acids composition was not influenced by the supplementation of lactoferrin. Peroxide value of matured T1, T2, T3, T4, and control was 0.31, 0.30, 0.28, 0.35, and 0.28 (meqO2/kg) with no variation in color, flavor and texture score. The results of the current investigation proved that lactoferrin can be used for the supplementation of cheddar cheese.
Nutritionists recommend to consume 3 g Conjugated linoleic acid (CLA) to prevent cancers, cardiovascular diseases, diabetes, obesity and other lifestyle-related disorders. To fulfill the daily requirements of CLA, large proportions of dairy and meat products have to be included in the dietary plan that may have negative impacts such as excessive intake of fat, cholesterol, and lactose. Current investigation was aimed to determine the effect of safflower oil derived CLA on fatty acids composition, lipolysis and sensory properties of cheddar cheese. Safflower oil-derived conjugated linoleic acid (CLA) was added to cheddar cheese at four levels i.e. 1%, 2%, 3% and 4% concentrations (T1, T2, T3 & T4), cheese without CLA supplement was used as control, vacuum packaged samples were ripened at 6–8°C for 90 days. Analysis of CLA supplement revealed that concentrations of ∆9c,11 t-18:2, ∆10 t,12c-18:2, ∆9c,11c-18:2, ∆9 t, 11c-18:2 were 52.16, 44.37, 0.27 and 0.55 mg/100 g. In control, concentrations of ∆9c,11 t-18:2 and ∆10 t,12c-18:2 was 0.24% and 0.07% with a total CLA content of 0.31%. In T3, concentrations of ∆9c,11 t-18:2, ∆10 t,12c-18:2, ∆9c,11c-18:2 and ∆9 t, 11c-18:2 were 1.12, 3.82, 1.63, 2.77 (total CLA 8.34mg100g). In T4, concentrations of ∆9c,11 t-18:2, ∆10 t,12c-18:2, ∆9c,11c-18:2 and ∆9 t, 11c-18:2, were 1.47, 3.50, 3.25 and 3.44% (total CLA 11.67 mg/100 g). Supplementation of cheddar cheese at all four levels significantly increased the concentrations of lactic acid, citric acid, propionic and acetic acids. After 90 days of ripening, free fatty acids in control, T1, T2, T3 and T4 were 0.14, 0.19, 0.23, 0.29 and 0.37% with no difference in composition and sensory properties. These results suggested that CLA content of cheddar cheese can be successfully increased with safflower oil-derived CLA supplement.
Abstract A total of 10 specimens were captured from selected sites of Bajaur Agency FATA, Pakistan using mist nets. The captured specimens were morphologically identified and various morphometric measurements were taken. The head and Body length (HB) of Pipistrellus coromondra and Pipistrellus kuhlii lepidus (n=10) was 43±0.11 mm and 45±1.1 respectively. Morphologically identified Pipistrellus kuhlii confirmed as Pipistrellus kuhlii lepidus based on 16S rRNA sequences. The DNA sequences were submitted to GenBank and accession numbers were obtained (MN 719478 and MT430902). The available 16S rRNA gene sequences of Pipistrellus coromondra and Pipistrellus kuhlii lepidus were retrieved from NCBI and incorporated in N-J tree analysis. Overall, the interspecific genetic variations among Pipistrellus coromondra and Pipistrellus kuhlii lepidus were 8% and 1% respectively. In our recommendation, a comprehensive molecular identification of bats is need of hour to report more cryptic and new species from Pakistan.
Aflatoxin B1 (AFB1) is the most hazardous toxin in food and feed components.Lactobacilli are extensively studied probiotic bacteria with detoxification characteristics.This study was conducted to isolate Lactobacillus from broiler gut, identify them by 16S ribosomal RNA (16S r RNA) gene sequence and evaluate their detoxification ability against AFB1.A total of fourteen Lactobacillus isolates were obtained from poultry gut and were identified.Phylogenetic study on the base of 16S rRNA gene showed that isolates from same species and source are genetically close to each other as compared to other isolates.Out of fourteen isolates, five have shown detoxification ability against AFB1 (50 ppb), in aqueous environment at 37 °C for 6 hours.Viable cells and cell walls have removed more concentration of AFB1 as compared to cell metabolites.Isolate Cr. 4 (L.salivarius) removed the highest amount (92.3%) of AFB1 as compared to other isolates.It was concluded that Lactobacillus isolates from broiler gut are potentially active detoxifying agents against AFB1 and their detoxification ability is strain dependent.
Buttermilk is a fermented drink widely used all over the world. It is produced widely both traditionally and commercially through pasteurization. Buttermilk, produced during churning of cultured cream, was supplemented with vitamin C at four different concentrations i.e., 100, 200, 300, 400 mg/100 ml, and zero concentrations (T1, T2, T3, T4 and control) and was evaluated for different parameters. Vitamin C supplementation at all concentrations had no effect on compositional attributes of buttermilk (p>0.05). The results also showed that concentration of vitamin C supplemented buttermilk in T1, T2, T3, T4, and control was 98.81, 196.14, 277.38, 382.19, and 0 mg/100 ml (p<0.05) respectively. It was also concluded that prolong storage period decreases vitamin C concentration. Concentration of lactic acid in control, T1, T2, T3 and T4 was 3047, 3019, 3027, 3062 and 3039 ppm (p>0.05) respectively. Oleic acid content in control, T1, T2, T3, T4 was 35.79, 35.75, 35.65, 35.63 and 35.19% (p>0.05). Linoleic acid content in control, T1, T2, T3, and T4 was 7.49, 7.05, 7.13, 7.10 and 7.33% (p<0.05) respectively. Addition of vitamin C supplemented buttermilk at all four concentrations had no effect on fatty acids composition (p>0.05) but the variation is due to storage period. Total antioxidant capacity of T1, T2, T3, T4 and control was 35.72, 42.74, 52.93, 68.77 and 84.66% (p<0.05). DPPH free radical scavenging activity of T1, T2, T3, T4 and control was 15.42, 24.87, 32.84, 40.37 and 49.78% (p<0.05) respectively. Free fatty acids and peroxide values of control and experimental samples were not different from each other (p>0.05). According to trained judges, sensory evaluation of all the treatments were performed on a 9 point hedonic scale and the results showed a non-significant difference between control and treatments. From commercial point of view, sensory results were promising as vitamin C did not affect sensory attributes of buttermilk which is quite acceptable from consumer perspective. It is therefore concluded from the results that buttermilk can be supplemented with vitamin C.
Abstract Lactobacilli ( n = 24), isolated from human infants and yogurt, showed variable in vitro activity against Salmonella enteritidis (8.0 ± 1.0 to 16.6 ± 0.5 mm) and other gut pathogens (9.0 ± 1.0 to 15.3 ± 0.5 mm), as determined by a well diffusion assay. The isolates were identified as Limosilactobacillus fermentum (FY1, FY3, FY4, IL2, and IL5), Lactobacillus delbrueckii (FY6 and FY7), Lactobacillus sp. (IL7), and Lactobacillus gasseri (IL12). All isolates showed variable in vitro tolerance to acidic pH for 3 h and visible growth at pH 4 and in the presence of 0.3% ox-bile. The antibiotic susceptibility profile of Lactobacillus isolates indicated resistance against vancomycin, ciprofloxacin, streptomycin, and lincomycin. Isolates had variable auto-aggregation and showed variable capabilities to co-aggregate with S. enteritidis . Based on all tested parameters, L. fermentum IL2, L. fermentum IL5, and L. gasseri IL12 were selected for co-culture experiments, followed by in vivo evaluation in Balb/c mice. All the selected isolates resulted in a 100% reduction in S. enteritidis in broth. Lactobacillus isolates efficiently colonized mouse guts and inhibited S. enteritidis colonization. Overall, there was ≥99.06% and ≤4.32 Mean log 10 reduction in Salmonella counts in mice feces within 7 days. The study, thus, provided characterized lactobacilli that could be considered as potential ingredients for probiotic formulations intended to prevent S. enteritidis infection in humans.
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