Draft genome sequencing of Enterococcus avium strains isolated from bovine mastitis.

We performed whole-genome sequencing of four multidrug-resistant Enterococcus avium strains isolated from milk (4M1), feces (4F1 and 4F2), and farm soil (4S1) of mastitic dairy cows. The draft genomes of E. avium strains 4M1, 4F1, 4F2, and 4S1 were approximately 4.2 Mbp, with 39.1% GC content and 66.5× coverage.

Draft genome sequence of Leuconostoc falkenbergense isolated from naturally fermented buffalo milk curd.

This study reports the draft genome of Leuconostoc falkenbergense strain BSMRAU-M1L5, isolated from artisanal buffalo milk curd in Bangladesh. The draft genome spans 1,776,471 bp, with 50× coverage and 96 contigs.

Draft genome sequencing of Pediococcus pentosaceus strains isolated from cow milk.

We sequenced the genomes of Pediococcus pentosaceus strains MBBL4 and MBBL6, isolated from raw milk samples of healthy cows. The draft genomes of the MBBL4 and MBBL6 were 1,896,831 bp and 1,849,397 bp, respectively, and were fragmented into 58 and 42 contigs, with coverages of 118.2× and 128.7×, respectively.

Genomic insights into antibiotic resistance genes in Leuconostoc citreum strains isolated from artisanal buffalo milk curd in Bangladesh through whole-genome sequencing.

We sequenced the genome of Leuconostoc citreum strains BSMRAU-M1L6 and BSMRAU-M1L13 isolated from artisanal buffalo milk curd in Bangladesh. The draft genomes of BSMRAU-M1L6 and BSMRAU-M1L13 are 1,869,891 and 1,890,611 bp, respectively, with 50.0× coverage (both) and 65 and 75 contigs, respectively.

Retardation of oxidative rancidity in ghee adding orange peel powder at different storage temperature.

This study is aimed to determine and compare the antioxidant activity of Orange Peel Powder (OPP) in ghee at different temperatures (4 °C, 25 °C and 60 °C) for divergent storage periods (0, 7, 14 and 21 days). To compare the antioxidant potentiality, synthetic antioxidant BHA (Butylated Hydroxy Anisole) is used. Twelve ghee samples were prepared where one was control, another one was BHA treated and the rest ten were admixing OPP in ghee at different ratios. After sensory evaluation three highest scored ghee samples (0.5%. 1.0% and 1.5%) were selected. Samples were analyzed for peroxide (PV), thiobarbituric acid (TBA), free fatty acids (FFA) value and radical scavenging activity. Though storage temperature and storage period were increased OPP treated ghee samples peroxide, TBA and FFA values were lowered significantly compared to control samples. Moreover, 1.0% and 1.5% OPP treated ghee samples such values were lowered than BHA treated ghee samples and all these are on the favor of ghee quality. OPP treated ghee samples' DPPH quench potentiality is also stronger than BHA treated ghee samples. Therefore, OPP is a great source of antioxidants and this can be used in ghee as a natural source of antioxidants.

Nutrient content and in vitro degradation study of some unconventional feed resources of Bangladesh.

This study aimed to evaluate six unconventional feed resources of Bangladesh, including water hyacinth (Eichhornia crassipes), banana leaves (Musa paradisiaca), roadside grass (Stenotaphrum secundatum), bamboo leaves (Bambusa vulgaris Scrad), Seaweed (Hypnea sp.) and sugarcane bagasse (Saccharum griffithii). Evaluations were based on dry matter (DM), crude protein (CP), crude fiber (CF), neutral detergent fiber (NDF), acid detergent fiber (ADF), ether extract (EE), ash content, DM and OM digestibilities and fractional rate of degradation. Two conventional feeds, i.e., rice bran and german grass, were used as the positive control. Samples (400 mg) were incubated with rumen liquor in an in vitro fermentation chamber at 0, 6, 12, 24, 48, 72, and 96 h for the degradation kinetic studies. The CP contents of 10.13, 10.63, 10.21, and 8.49 % were found in seaweed, banana leaf, water hyacinth, and bamboo leaf, respectively. The NDF values ranged between 16.5 and 75.6% and ADF varied from 9.7 to 58.8% in this study. The highest value of NDF (75.6%) and ADF (58.8%) were found in sugar cane bagasse and the lowest value of NDF (16.5%) and ADF (9.7%) were as observed in seaweed. However, higher DM degradation (33.5-42.8%) was found in seaweed during the incubation periods of 24-96 h. A significant (P < 0.05) increased of OM degradation (44.9%) compared to other feed resources was also observed in seaweed at 96 h of in vitro incubation. Water hyacinth, banana leaves, german grass, and sugarcane bagasse had greater DM digestibility (32.9-36.3%) compared to roadside grass, bamboo leaves, and rice bran (24.8-29.1%). The higher total OM digestibility of seaweed found (>44.9%) can be associated with the presence of large quantities of fraction b (>39.2 %), resulting in moderate amounts of undegradable fraction (U) (57.2 %). This study provides a comparative estimate of ruminal DM and OM degradation characteristics for seaweed and some other unconventional feed resources, which might be helpful for their inclusion in the diet according to the ruminally undegraded to degraded DM and OM intake ratio.

Assessment of ghee adulterated with oils and fats in Bangladesh.

OBJECTIVE: This study aimed at determining the existence of oils and fats in ghee manufactured in Bangladesh and to validate the nature of the impurity. MATERIALS AND METHODS: In this study, a ghee sample was prepared in the laboratory by following standard methods and was used as a control sample. On the other hand, 19 ghee samples, including five branded samples (B1-B5), and 14 local samples (L1-L14) were collected from different manufacturers. The ghee samples were assessed for fat composition, Reichert Meissl (RM), saponification, Polenske, acid, Kirschner, and butyro refractometer (BR) values. To validate the ghee samples, vegetable oils and body fats were mixed in different ratios and then analyzed. RESULTS: All the branded samples contained more than 99.5% fat, but only three local samples showed more than 97% fat. Admixing of soybean oil and coconut oil in different ratios showed the RM value from 1.57 ± 0.09 to 4.14 ± 0.21, whereas incorporation of hydrogenated vegetable oils and tallow showed 6.36 ± 0.03 to 14.10 ± 0.14. Nine local samples revealed RM values similar to external fat admixed samples. B2, B4, B5, L2-L8, and L10-L14 samples' saponification values differed from the standard limits. Polenske, acid, Kirschner values and BR reading for L4, L6, L7, L8, L10, L12, L13, and L14 showed the worst results. All values varied significantly (p < 0.01). CONCLUSION: Local samples, L4, L6, L7, L8, L10, L12, L13, and L14, were assumed to be adulterated with external oils and fats. The quality of local ghee is questionable, as the samples contained more than 8% moisture, whereas pure ghee had less than 0.5% moisture.

Dyeing of Polyester with 4-Fluorosulfonylphenylazo-5-pyrazolone Disperse Dyes and Application of Environment-Friendly Aftertreatment for Their High Color Fastness.

Dyeing and fastness properties of a series of 4-fluorosulfonylphenylazo-5-pyrazolone dyes on polyester were investigated in this study. The 4-nitrophenylazo-5-pyrazolone dyes were also synthesized to compare their dyeing and fastness properties on polyester with those of fluorosulfonyl-substituted analogues. The substantivity of 4-arylazo-5-pyrazolone derivatives containing a p-fluorosulfonyl group in the diazo component was lower than that of their nitro analogues which have a higher extinction coefficient and higher affinity because of the polar nitro group. They showed relatively hypsochromic color and lower chroma on polyester compared with their nitro analogues because of the relatively weaker electron-accepting power of the fluorosulfonyl group compared to the nitro group. Disperse dyeing of polyester with 4-fluorosulfonylphenylazo-5-pyrazolone disperse dyes achieved high color fastness and reduces the adverse environmental impact of the dyeing process by providing the option of performing alkali clearing instead of reductive clearing, which has high biological oxygen demand when discharged into the dyeing effluent and generates carcinogenic aromatic amines.

Underlying mechanisms involved in the decrease of milk secretion during Escherichia coli endotoxin induced mastitis in lactating mice.

Mastitis, the inflammation of mammary glands resulting from bacterial infection, disrupts milk production in lactating mammary glands. In this study, we injected lipopolysaccharide (LPS), one of the endotoxins from Escherichia coli into mouse mammary glands to disrupt milk production, and we investigated the influence of LPS on nutrient uptake, synthesis, and secretion processes for milk component production in alveolar epithelial cells (AEC). The expression of genes relevant to the three-staged milk component production process (nutrient uptake, synthesis, and secretion of milk components) were down-regulated within 12 h after LPS injection in AEC. The internalization of glucose transporter 1 (GLUT-1) from the basolateral membrane to the cytoplasm occurred in accordance with the down-regulation of gene expression 3 h after LPS injection. The abnormal localization of adipophilin and beta-casein was also observed in the LPS-injected mammary glands. SLC7A1, an amino acid transporter, was up-regulated 3 and 6 h after LPS injection. Furthermore, the inactivation of signal transducer and activator of transcription 5 (STAT5) and the activation of STAT3 and nuclear factor-kappa B (NFkappaB) occurred 3 h after LPS injection. These results indicate that the nutrient uptake, synthesis, and secretion of milk components in AEC are rapidly shut down in the lactating mammary glands after LPS injection.

Histological analysis of mammary gland remodeling caused by lipopolysaccharide in lactating mice.

The mammary alveolus is a highly specialized structure that secretes milk for suckling infants during lactation. The secreting alveolus consists in alveolar epithelial cells (AECs) and myoepithelial cells and is surrounded by microvascular endothelial cells, adipocytes and several immune cell types such as macrophages and neutrophils. During normal lactation, these cells play distinct roles needed to maintain the secretory ability of the mammary alveolus. However, inflammation resulting from pathogenic bacterial infections causes structural and functional regression of the secreting alveolus in the lactating mammary gland. We initiated artificial inflammation in the mammary glands of lactating mice by injecting lipopolysaccharide (LPS), as a mammary inflammation model and investigated, by immunohistochemical analysis, the early response of the cells constituting and surrounding the alveolus. Some AECs sloughed away from the alveolar epithelial layer and showed progression of apoptosis detected by immunostaining of cleaved caspase-3 after LPS injection. Adipocytes exhibited transient shrinkage and re-accumulation of lipid droplets, although the numbers of adipocytes did not demonstrate a significant difference. Activation of F4/80-positive cells around the mammary alveolus was observed 3 h after LPS injection. However, the recruitment of CD11b-positive cells into the alveolar lumen was not observed until 12 h after LPS injection. Myoepithelial cells were contracted after LPS injection. LPS injection around the alveolus did not induce any detectable structural changes in capillaries surrounding the alveolus. Thus, cell-specific behavior and tissue remodeling of the alveolus occur after LPS injection in a time-dependent manner.

Lipopolysaccharide disrupts the milk-blood barrier by modulating claudins in mammary alveolar tight junctions.

Mastitis, inflammation of the mammary gland, is the most costly common disease in the dairy industry, and is caused by mammary pathogenic bacteria, including Escherichia coli. The bacteria invade the mammary alveolar lumen and disrupt the blood-milk barrier. In normal mammary gland, alveolar epithelial tight junctions (TJs) contribute the blood-milk barrier of alveolar epithelium by blocking the leakage of milk components from the luminal side into the blood serum. In this study, we focused on claudin subtypes that participate in the alveolar epithelial TJs, because the composition of claudins is an important factor that affects TJ permeability. In normal mouse lactating mammary glands, alveolar TJs consist of claudin-3 without claudin-1, -4, and -7. In lipopolysaccharide (LPS)-induced mastitis, alveolar TJs showed 2-staged compositional changes in claudins. First, a qualitative change in claudin-3, presumably caused by phosphorylation and participation of claudin-7 in alveolar TJs, was recognized in parallel with the leakage of fluorescein isothiocyanate-conjugated albumin (FITC-albumin) via the alveolar epithelium. Second, claudin-4 participated in alveolar TJs with claudin-3 and claudin-7 12 h after LPS injection. The partial localization of claudin-1 was also observed by immunostaining. Coinciding with the second change of alveolar TJs, the severe disruption of the blood-milk barrier was recognized by ectopic localization of ß-casein and much leakage of FITC-albumin. Furthermore, the localization of toll-like receptor 4 (TLR4) on the luminal side and NFκB activation by LPS was observed in the alveolar epithelial cells. We suggest that the weakening and disruption of the blood-milk barrier are caused by compositional changes of claudins in alveolar epithelial TJs through LPS/TLR4 signaling.

Target of rapamycin (TOR) controls vitellogenesis via activation of the S6 kinase in the fat body of the tick, Haemaphysalis longicornis.

Vitellogenin (Vg) synthesis, vitellogenesis, is an essential process for the development and reproduction of ticks. Our previous finding led to the hypothesis that target of rapamycin (TOR) pathway is important for vitellogenesis in the hard tick, Haemaphysalis longicornis. The TOR pathway controls cellular activity according to nutrient availability in eukaryotes. TOR, a member of the phosphatidylinositol 3-kinase family, is a central player in this pathway. Here, we present preliminary evidence that H. longicornis TOR (HlTOR) controls vitellogenesis via activation of S6 kinase (S6K) in the fat body. RNA interference (RNAi)-mediated gene silencing of HlTOR was undertaken to elucidate the involvement of HlTOR in the vitellogenesis of the tick. HlTOR-RNAi caused inhibition of S6K phosphorylation in the fat body. HlTOR-RNAi also altered not only the expression levels of GATA mRNA and protein but also the intracellular localisation of GATA in the fat body. The expression levels of Vg mRNA and protein in the fat body of HlTOR-RNAi ticks were significantly lower than those in control ticks. In the pre-ovipositional stage, the ovaries of control ticks had brown oocytes developing, but those of HlTOR-RNAi ticks were white and immature. The haemolymph colour indicated that the amount of Vg was lower in HlTOR-RNAi ticks than in the controls. Furthermore, rapamycin inhibited S6K phosphorylation and reduced the expression levels of Vg mRNA and protein in the fat bodies. Vg proteins were not detected in rapamycin-treated fat bodies in the presence of 20-hydroxyecdysone. These results suggest that HlTOR activity is critical for vitellogenesis stimulated by 20-hydroxyecdysone.

Parasiticidal activity of human alpha-defensin-5 against Toxoplasma gondii.

Human defensins play a fundamental role in the initiation of innate immune responses to some microbial pathogens. In this paper, we show that human alpha-defensin-5 displays a parasiticidal role against Toxoplasma gondii, the causative agent of toxoplasmosis. Exposure of the tachyzoite form of T. gondii to defensin induced aggregation and significantly reduced parasite viability in a concentration-dependent peptide. Pre-incubation of tachyzoites with human alpha-defensin-5 followed by exposure to a mouse embryonal cell line (NIH/3T3) significantly reduced T. gondii infection in these cells. Thus, human alpha-defensin-5 is an innate immune molecule that causes severe toxocity to T. gondii and plays an important role in reducing cellular infection. This is the first report showing that human alpha-defensin-5 causes aggregation, leading to Toxoplasma destruction.

The identification and characterization of lysozyme from the hard tick Haemaphysalis longicornis.

A full-length cDNA-encoding lysozyme was obtained from cDNA libraries of salivary glands of the hard tick Haemaphysalis longicornis and designated as HlLysozyme. The HlLysozyme sequence represents an open reading frame for a putative signal peptide and the mature protein composed of 121 amino acids. The calculated molecular weight of the protein is 13.7 kDa, and the theoretical isoelectric point is 9.85. HlLysozyme shares 41-79% amino acid sequence identity with the lysozymes of other organisms. The activity of recombinant HlLysozyme expressed in Escherichia coli was confirmed by a lytic zone assay using lyophilized Micrococcus lysodeikticus. The HlLysozyme activity decreased at 70 °C and was demonstrated at acidic side and neutral in a pH range. Elevated gene expression of HlLysozyme was observed when female ticks were challenged with bacteria, suggesting possible roles of lysozyme as an innate immunity of ticks against microorganisms.

Growth promotion and cell binding ability of bovine lactoferrin to Bifidobacterium longum.

Lactoferrin, a major whey protein of human milk, is considered as growth promoter for bifidobacteria, the predominant microorganisms of human intestine. In the present study, in vitro growth promotion and cell binding ability of bovine lactoferrin to several strains of Bifidobacterium longum have been demonstrated. A dose-dependent as well as strain-dependent growth promotion effect by lactoferrin was observed. Cell binding ability of lactoferrin was inspected under an inverted confocal laser scanning microscope by incubation bacterial cells with biotinylated bovine lactoferrin and FITC-conjugated avidin. Fluorescence staining showed bovine lactoferrin binding to all tested strains. A lactoferrin-binding protein with a molecular weight of approximately 67 kDa was also detected in the extracted membrane and cytosolic fraction of each B. longum strain by far-Western blot technique using biotinylated lactoferrin and horseradish peroxidase-conjugated streptavidin. Based on these results, we suggest that existence of lactoferrin-binding protein could be a common characteristic in bifidobacteria. It can also be hypothesized that lactoferrin-binding protein in bifidobacteria is not only involved in growth stimulation mechanism but also could play different roles.