Fatty acid and amino acid profiles of cheese, butter, and ghee made from buffalo milk.

Objective: The objective was to assess the chemical composition, cholesterol, fatty acid (FAs), and amino acid (AAs) profiles of buffalo cheese, butter, and ghee. Materials and Methods: Buffalo milk (raw) was collected from the Bangladesh Agricultural University (BAU) Dairy Farm, BAU, Mymensingh-2202, Bangladesh. Cheese, butter, and ghee were prepared at the Dairy Chemistry and Technology Laboratory, Department of Dairy Science, BAU, Mymensingh, Bangladesh, and subjected to subsequent analyses. The gross nutritional composition and AAs profile of milk were analyzed prior to the manufacture of cheese, butter, and ghee. The gross nutritional composition of milk and dairy products was analyzed by applying an automated milk analyzer and the Association of Agricultural Chemists techniques, respectively. The cholesterol, FAs, and AAs contents of cheese, butter, and ghee were determined by the Bangladesh Council for Scientific and Industrial Research, Dhaka, Bangladesh. Furthermore, atherogenic and thrombogenic indices were also calculated using reference equations. Results: The results indicated that the buffalo milk is a good source of first-rate nutrients (dry matter: 16.50%, fat: 7.50%, protein: 3.75%). Findings indicated that the butter was significantly rich with (p < 0.05) total solids and fat where higher (p > 0.05) protein, carbohydrate, and minerals were found in cheese. The saponification, Reichert-Meissl, Polenski, and Kirschner values of buffalo ghee were found to be 225, 30, 1.2, and 25, respectively. A significant (p < 0.05) variation was found in the cholesterol content of buffalo cheese, butter, and ghee. Butter and ghee had 40.14 and 39.57 mg more cholesterol, respectively, than cheese. The results revealed identical FA profiles except for C24:0 among the three dairy products where the major FA compositions were C4:0, C14:0, C16:0, and C18:0 and C18:1 cis-9. The atherogenicity index and thrombogenicity index of cheese, butter, and ghee were statistically similar (p > 0.05). Butter was found with the most conducive anti-atherogenic and anti-thrombogenic characteristics due to lower saturated and higher polyunsaturated FAs. However, all the AAs concentrations were statistically higher (p < 0.05) in cheese than in butter and ghee. Conclusion: To conclude, buffalo cheese is superior to butter and ghee as regards nutrient density, but consumers can choose other foods based on their choice.

Physicochemical characteristics, sensory profile, probiotic, and starter culture viability of synbiotic yogurt.

Objectives: This study aimed to envisage the effectiveness of adding three particular prebiotics (inulin, ß-glucan, and Hi-maize) to synbiotic yogurt's physicochemical properties, sensory characteristics, and survivability of the probiotic and starter cultures. Materials and Methods: The yogurt's gross composition, syneresis, water-holding capacity (WHC), viscosity, sensorial properties, and probiotic and starter cell stability were analyzed. The Lactobacillus delbrueckii subsp. bulgaricus M240-5 and Streptococcus thermophilus M140-2 were employed as yogurt starter bacteria, and Lactobacillus acidophilus LA-5 as probiotic culture. The synbiotic yogurt was formulated with 5% sucrose and 0.7% artificial vanilla flavor. Results: The findings showed that when prebiotic ingredients were added to synbiotic yogurt, it had a significant impact on its sensory qualities, WHC, syneresis, and viscosity when compared to plain yogurt samples. The prebiotics did not affect the pH and titratable acidity of the yogurt samples. Additionally, the prebiotic supplementation did not influence the protein and fat content of synbiotic yogurt (p < 0.05). Prebiotics had an impact on the probiotic cell viability and total viable count (p < 0.05) compared to the plain sample, the 2.5% ß-glucan, 1.5% and 2.5% Hi-maize samples had the highest mean viability (8.95 Log CFU/ml). The starter culture ratio remained stable in response to the prebiotic levels. Conclusion: In summary, the production of synbiotic yogurts supplemented with Hi-maize and ß-glucan at 1.5% and 2.5%, respectively, is highly advised because these supplementations provide yogurt with acceptable syneresis, viscosity, WHC, and sensory attributes.

Mercury (II) sensor based on monitoring dissociation rate of the trans-acting factor MerR from cis-element by surface plasmon resonance.

Transcriptional switches regulate gene expression in response to environmental changes surrounding cell. Many studies have focused on two fundamentally different models of transcriptional control by bacterial metalloregulatory protein. Distortion of the DNA fragment including cis-element, to which the trans-acting factor MerR binds, is accepted as the mechanism of gene expression regulation by Hg (II) while, in cases of the other trans-acting factors ArsR and CadC, events of association to and dissociation from cis-element are known to control transcription in response to As (III) and Cd (II), respectively. In this study, interactions between green-fluorescent-protein-tagged trans-acting factor and immobilized cis-element were analyzed on solid surface. Fluorescent measurements and surface plasmon resonance (SPR) responses revealed that although the equilibrium dissociation constant (KD) was much lower in MerR than in ArsR and CadC, the dissociation rate of MerR from DNA increased in response to Hg (II) at concentrations of 5-10(4) µg l(-1). These results firstly demonstrate an increase of KD between MerR and its recognition site in DNA by Hg (II), and possibility of rapid Hg (II) quantification with the low detection limit (5 µg l(-1)) and the high dynamic range (10(1)-10(4) µg l(-1)).

Population abundance of potentially pathogenic organisms in intestinal microbiome of jungle crow (Corvus macrorhynchos) shown with 16S rRNA gene-based microbial community analysis.

Jungle Crows (Corvus macrorhynchos) prefer human habitats because of their versatility in feeding accompanied with human food consumption. Therefore, it is important from a public health viewpoint to characterize their intestinal microbiota. However, no studies have been involved in molecular characterization of the microbiota based on huge and reliable number of data acquisition. In this study, 16S rRNA gene-based microbial community analysis coupled with the next-generation DNA sequencing techniques was applied to the taxonomic classification of intestinal microbiome for three jungle crows. Clustering of the reads into 130 operational taxonomic units showed that at least 70% of analyzed sequences for each crow were highly homologous to Eimeria sp., which belongs to the protozoan phylum Apicomplexa. The microbiotas of three crows also contained potentially pathogenic bacteria with significant percentages, such as the genera Campylobacter and Brachyspira. Thus, the profiling of a large number of 16S rRNA gene sequences in crow intestinal microbiomes revealed the high-frequency existence or vestige of potentially pathogenic microorganisms.

Thermoresponsive magnetic nano-biosensors for rapid measurements of inorganic arsenic and cadmium.

Green fluorescent protein-tagged sensor proteins, ArsR-GFP and CadC-GFP, have been produced as biosensors for simple and low-cost quantification of As(III) or Cd(II). In this study, the sensor protein-promoter DNA complexes were reconstructed on the surfaces of magnetic particles of different sizes. After the surface modification all the particles could be attracted by magnets, and released different amounts of GFP-tagged protein, according to the metal concentrations within 5 min, which caused significant increases in fluorescence. A detection limit of 1 µg/L for As(III) and Cd(II) in purified water was obtained only with the nanoparticles exhibiting enough magnetization after heat treatment for 1 min. Therefore, thermoresponsive magnetic nano-biosensors offer great advantages of rapidity and sensitivity for the measurement of the toxic metals in drinking water.

Fluorescent bioassays for toxic metals in milk and yoghurt.

BACKGROUND: From a human health viewpoint, contaminated milk and its products could be a source of long-term exposure to toxic metals. Simple, inexpensive, and on-site assays would enable constant monitoring of their contents. Bioassays that can measure toxic metals in milk or yoghurt might reduce the risk. For this purpose, the green fluorescent protein (GFP)-tagged trans factors, ArsR-GFP and CadC-GFP, together with their cis elements were used to develop such bioassays. RESULTS: ArsR-GFP or CadC-GFP, which binds either toxic metal or DNA fragment including cis element, was directly mixed with cow's milk or yoghurt within a neutral pH range. The fluorescence of GFP, which is reflected by the association/dissociation ratio between cis element and trans factor, significantly changed with increasing externally added As (III) or Cd (II) whereas smaller responses to externally added Pb (II) and Zn (II) were found. Preparation and dilution of whey fraction at low pH were essential to intrinsic zinc quantification using CadC-GFP. Using the extraction procedure and bioassay, intrinsic Zn (II) concentrations ranging from 1.4 to 4.8 mg/l for milk brands and from 1.2 to 2.9 mg/kg for yoghurt brands were determined, which correlated to those determined using inductively coupled plasma atomic emission spectroscopy. CONCLUSIONS: GFP-tagged bacterial trans factors and cis elements can work in the neutralized whole composition and diluted whey fraction of milk and yoghurt. The feature of regulatory elements is advantageous for establishment of simple and rapid assays of toxic metals in dairy products.

Solid phase biosensors for arsenic or cadmium composed of A trans factor and cis element complex.

The presence of toxic metals in drinking water has hazardous effects on human health. This study was conducted to develop GFP-based-metal-binding biosensors for on-site assay of toxic metal ions. GFP-tagged ArsR and CadC proteins bound to a cis element, and lost the capability of binding to it in their As- and Cd-binding conformational states, respectively. Water samples containing toxic metals were incubated on a complex of GFP-tagged ArsR or CadC and cis element which was immobilized on a solid surface. Metal concentrations were quantified with fluorescence intensity of the metal-binding states released from the cis element. Fluorescence intensity obtained with the assay significantly increased with increasing concentrations of toxic metals. Detection limits of 1 µg/L for Cd(II) and 5 µg/L for As(III) in purified water and 10 µg/L for Cd(II) and As(III) in tap water and bottled mineral water were achieved by measurement with a battery-powered portable fluorometer after 15-min and 30-min incubation, respectively. A complex of freeze dried GFP-tagged ArsR or CadC binding to cis element was stable at 4 °C and responded to 5 µg/L As(III) or Cd(II). The solid phase biosensors are sensitive, less time-consuming, portable, and could offer a protocol for on-site evaluation of the toxic metals in drinking water.

Application of fluorescent protein-tagged trans factors and immobilized cis elements to monitoring of toxic metals based on in vitro protein-DNA interactions.

Environmental toxic metals cause serious global public health problems. On-site monitoring protects people from exposure to such harmful elements. In this study, the bacterial transcriptional switches were applied to monitoring of toxic metals. ArsR and CadC, trans factors of Escherichia coli and Staphylococcus aureus, were fused to GFP. The fusion proteins, ArsR-GFP and CadC-GFP, associated with cis elements, P(ars)-O(ars) and P(cad)-O(cad), respectively and dissociated from those upon recognition of As(III) or Pb/Cd. Cell lysates containing ArsR-GFP were pre-incubated with As(III) standard solutions for 15 min and loaded into P(ars)-O(ars)-immobilized microplate wells. Cell lysates containing CadC-GFP were pre-incubated with Pb or Cd solutions and loaded into P(cad)-O(cad)-immobilized wells. The cell lysates were incubated for 15 min and removed from the wells. Fluorescence intensity in the wells dose-dependently decreased in response to As(III) up to 200 µg/l or Pb/Cd up to 100 µg/l. Detection limits were 10 µg/l for As(III) 10 µg/l for Cd, and 20 µg/l for Pb with a microplate fluororeader, whereas 5.0 µg/l for As(III), 1.0 µg/l for Cd, and 10 µg/l for Pb with a handheld fluorometer. This method was available to detect Pb/Cd or As(III) in water containing soil extracts. This is the first demonstration of a simple and rapid fluorometry to detect analytes based on in vitro interaction between a cis element and a trans factor.