Construction of a Salmonella Gallinarum ghost as a novel inactivated vaccine candidate and its protective efficacy against fowl typhoid in chickens

In order to develop a novel, safe and immunogenic fowl typhoid (FT) vaccine candidate, a Salmonella Gallinarum ghost with controlled expression of the bacteriophage PhiX174 lysis gene E was constructed using pMMP99 plasmid in this study. The formation of the Salmonella Gallinarum ghost with tunnel formation and loss of cytoplasmic contents was observed by scanning electron microscopy and transmission electron microscopy. No viable cells were detectable 24 h after the induction of gene E expression by an increase in temperature from 37 °C to 42 °C. The safety and protective efficacy of the Salmonella Gallinarum ghost vaccine was tested in chickens that were divided into four groups: group A (non-immunized control), group B (orally immunized), group C (subcutaneously immunized) and group D (intramuscularly immunized). The birds were immunized at day 7 of age. None of the immunized animals showed any adverse reactions such as abnormal behavior, mortality, or signs of FT such as anorexia, depression, or diarrhea. These birds were subsequently challenged with a virulent Salmonella Gallinarum strain at 3 weeks post-immunization (wpi). Significant protection against the virulent challenge was observed in all immunized groups based on mortality and post-mortem lesions compared to the non-immunized control group. In addition, immunization with the Salmonella Gallinarum ghosts induced significantly high systemic IgG response in all immunized groups. Among the groups, orally-vaccinated group B showed significantly higher levels of secreted IgA. A potent antigen-specific lymphocyte activation response along with significantly increased percentages of CD4⁺ and CD8⁺ T lymphocytes found in all immunized groups clearly indicate the induction of cellular immune responses. Overall, these findings suggest that the newly constructed Salmonella Gallinarum ghost appears to be a safe, highly immunogenic, and efficient non-living bacterial vaccine candidate that protects against FT.     

Extract of Magnoliae Flos inhibits ovariectomy-induced osteoporosis by blocking osteoclastogenesis and reducing osteoclast-mediated bone resorption

Bone homeostasis is maintained by a balance between bone resorption by osteoclasts and bone formation by osteoblasts. Osteoporosis occurs when osteoclast activity surpasses osteoblast activity. Pro-inflammatory cytokines stimulate osteoclast differentiation and activity by increasing production of macrophage-colony stimulating factor and receptor activator of nuclear factor-κB ligand (RANKL). In this study, we investigated whether Magnoliae Flos (MF), one of the most commonly used Chinese medicinal herbs for managing rhinitis, sinusitis and headache, could effectively inhibit osteoporosis. In ovariectomized (OVX) mice compared to sham mice, the body weight increased and serum levels of alkaline phosphatase (ALP), tartrate resistant acid phosphatase 5b, calcium, and osteocalcin were significantly elevated. However, orally administrated MF extract substantially inhibited the increased body weight and serum levels of bone turnover markers, without any evidence of tissue toxicity. MF extract treatment significantly reversed the morphometric parameters of ovariectomy-induced bone loss, including trabecular bone volume, thickness, number, separation, and bone density, to almost the same levels of the sham mice. Furthermore, MF extract reduced the RANKL-mediated osteoclast differentiation and bone resorption by inhibiting the activities of matrix metalloproteinases (MMPs) and cathepsin K in mouse bone marrow macrophages. MF extract appeared to increase ALP activity in murine osteoblastic cells. Taken together, MF extract may be a beneficial supplement for the blockade of osteoporosis progression, particularly for the management of postmenopausal osteoporosis.     

Comparative mutant prevention concentration and mechanism of resistance to veterinary fluoroquinolones in Staphylococcus pseudintermedius

Background – The problem of antibacterial drug resistance is increasing worldwide, in part due to the therapeutic concentrations currently used based on the minimal inhibitory concentration (MIC) as a measure of potency are often the very concentrations required to selectively enrich the resistant mutant portion of the population. A mutant prevention concentration (MPC)‐based dosing strategy is suggested to improve the therapeutic outcome based on the MIC. Objective – Our aim was to investigate the MPC and mechanism of resistance to various fluoroquinolones using recent Staphylococcus pseudintermedius isolates from canine pyoderma. Methods – The broth microdilution method for MIC and a series of agar plates containing different concentrations of fluoroquinolones were inoculated with ～10¹⁰ colony‐forming units of the bacterial culture for MPC were used. PCR was used to identify mutation in the resistant isolates. Results – The rank order of potency based on MIC and MPC was ciprofloxacin = enrofloxacin ≥ marbofloxacin > difloxacin ≥ orbifloxacin. Integrating our data with reported pharmacokinetic data at the recommended dose ranges revealed that only high doses of ciprofloxacin, enrofloxacin and marbofloxacin could achieve a maximal plasma concentration (C_max) greater than the MPC of 90% of isolates (C_max/MPC₉₀). The overall rank of potency against S. pseudintermedius, based on C_max/MIC, C_max/MPC, the area under concentration–time curve (AUC)/MIC and AUC/MPC values, was in decreasing order: enrofloxacin > ciprofloxacin ≥ marbofloxacin ≥ orbifloxacin = difloxacin. Sequencing of the quinolone resistant determining region of gyrA, gyrB, grlA and grlB of resistant strains showed a base‐pair substitution in both gyrA and gyrB that resulted in Ser‐84 to Leu and Ser‐80 to Arg amino acid changes, respectively. Conclusions and clinical importance – High doses of ciprofloxacin, enrofloxacin and marbofloxacin could minimize the selection of resistant mutants, whereas the possibility of selecting mutants with the conventional doses of difloxacin and orbifloxacin, and low clinical doses of all fluoroquinolones, seems high.     

Molecular epidemiologic investigation of lentogenic Newcastle disease virus from domestic birds at live bird markets in Korea

A Newcastle disease surveillance program was conducted at live bird markets in Korea to expand our epidemiologic understanding of the disease in Korea. During the surveillance program, 10 lentogenic Newcastle disease viruses (NDVs) were isolated and identified from apparently healthy chickens and ducks at live bird markets. The lentogenic viruses had sequence motifs of either 112GKQGRL117 (n = 8) or 112GRQGRL117 (n = 2) at the F0 cleavage site. Sequencing and phylogenetic analyses of NDV isolates based on the hypervariable region of the F protein revealed two different genotypes: genotypes I (n = 8) and II (n = 2). Genotype I viruses were most closely related to the NDV V4 strain (n = 7) or the NDV Ulster 2C strain (n = 1). In contrast, genotype II viruses clustered with the NDV vaccine strains (LaSota and VG/GA) that are commonly used as live vaccines in Korea. The epidemiologic importance of NDV at live bird markets in Korea is discussed.     

Flower bud abscission triggered by the anther in the Asiatic hybrid lily

It is not well documented which floral parts may trigger the onset of tepal or petal senescence and or flower bud abscission. Asiatic hybrid (Lilium × elegans Thunb.) ‘Red Carpet’ lily flowers were selected as a model to study this relationship because the various floral organs of a lily flower can be easily dissected and collected for physiological studies. We hypothesized that anther is the organ that triggers flower bud abscission. Ethanol-soluble sucrose, fructose, and glucose were analyzed in the tepal, anther, filament, and pistil. The analysis was conducted on flower buds weighing 1.5 g (small buds) and 3.9 g (large buds). The sucrose content in the anthers of small buds remained high when cut flowers were held at 21 °C for 5 d, suggesting that sucrose is not hydrolyzed into fructose and glucose. However, in large buds, sucrose was not detected when flowers were held at 21 °C for 5 d. The respiration rate of small buds (2.06 g) was high (2000-3000 mg/kg fw) when the tepals were removed and the anther remained attached to the filament, but was low (less than 400 mg/kg fw) in large buds approaching anthesis. Ethylene was produced only in small buds (1.42 g) when the anthers were present. Radioactive ¹⁴C sucrose applied to the cut end of the pedicel was translocated rapidly to the anther at harvest, but was not translocated to the anther when abscission was induced by keeping flower buds at 21 °C. Ethylene detected in small flower buds possibly was stimulated by physiological changes resulting from the lack of assimilates available to the anther and the lack of sucrose hydrolysis occurring in the anther. The absence of sucrose hydrolysis in the anther and lack of translocation of sucrose from the filaments to the anther in small buds may be factors that contribute to flower bud abscission in ‘Red Carpet’ lily.     

Water-resistant Lignin/Poly(vinyl alcohol) Blend Fibers for Removal of Hexavalent Chromium

Lignin is the second most abundant renewable biomass-derived natural resource that has been used to replace traditional petrochemical-based materials. However, fabricating the lignin component into the various forms required for practical application is still challenging. In this work, we fabricated water-resistant lignin/poly(vinyl alcohol) (PVA) blend fibers by wet spinning and glutaraldehyde crosslinking methods. The effect of the lignin/PVA blend ratio and glutaraldehyde crosslinking process on the physicochemical properties of wet-spun lignin/PVA blend fibers were studied using maximum draw ratios, hydrolytic degradation profiles, and mechanical properties. Furthermore, the hexavalent chromium [Cr(VI)] removal behavior of lignin/PVA blend fibers was investigated according to the effect of pH, initial Cr(VI) concentration, and contact time. The wet-spun lignin/PVA blend fiber achieved excellent water stability through glutaraldehyde crosslinking and exhibited notable Cr(VI) adsorption capacity (350.87 mg/g) and good regeneration ability. These findings demonstrate that glutaraldehyde-crosslinked lignin/PVA blend fibers could be promising adsorbents for the remediation of heavy metal species containing textile wastewater.     

High Light-Fastness Acid Dyes Synthesized from Corresponding Anthraquinone Chromophore Utilizing a Sulfonation Reaction. I. Dye Synthesis and Characterization

As the diameter of polyamide fibers decreased to finer denier, the dyeing fastness tends to be deteriorated due to the increase of their surface area, particularly both light fastness and wash fastness. In this study, three acid dyes were synthesized utilizing a sulfonation reaction starting from corresponding hydrophobic dye (for yellow and red dye) and dye intermediate containing a sulfonic acid group (for blue dye), those featured by high light fastness property. A Gaussian structural prediction model was used to determine the structure of the acid dyes prior to dye synthesis, and the optimal structures of three acid dyes were analyzed. Dye structures prepared were confirmed by ¹H-NMR, mass spectroscopy, and elemental analysis. By using a UV-vis spectrophotometer, the absorption maxima and molar extinction coefficient were also measured for three acid dyes comparing to that of the corresponding disperse dye or blue dye intermediate. Judging from spectroscopic data, the introduction of sulfonyl groups led to increase of molar extinction coefficient and a bathochromic shift.     

Cholecystokinin mediates depression of feed intake in dairy cattle fed high fat diets

Two experiments were conducted to investigate the effects of 1) different concentrations of dietary fat and 2) i.v. administration of a cholecystokinin receptor antagonist (MK-329) on feed intake and plasma concentrations of hormones and metabolites in dairy cattle. In Experiment 1, 4 lactating Holstein cows were used in a 4 x 4 Latin square design. Treatments were diets with 1) no fat added, 2) 30 g fat/kg feed (calcium salts of long-chain fatty acids as fat supplement), 3) 60 g fat/kg, and 4) 90 g fat/kg added. Cows were fed once daily a diet of concentrate, corn silage, alfalfa haylage, and alfalfa pellets. Dry matter intake decreased linearly with increasing concentrations of dietary fat (P < 0.0001). Overall plasma concentrations of nonesterified fatty acids (P < 0.0001), triacylglycerol (P < 0.0006), and cholecystokinin (P < 0.02), increased linearly with each level of dietary fat, but there was a linear decrease in plasma insulin (P < 0.0008). In Experiment 2, 4 nonpregnant and nonlactating Holstein heifers were used in a cross-over design in a 2 x 2 factorial arrangement of treatments. Treatments were diet (fatty acids, 27 g/kg vs 103 g/kg diet dry matter) and i.v. injections (MK-329 vs vehicle). Heifers were fed once daily a total mixed ration of corn silage, cracked corn and soybean meal with or without fat supplement. Diets were switched by period and either MK-329 (70 microg/kg body weight) or its vehicle was injected i.v. at 2 hr postfeeding. Daily dry matter intake was decreased by feeding the high fat diet (P < 0.02) but was not affected by injections. Injection of MK-329, however, increased dry matter intake by 92% in heifers fed the high fat diet during the first 2 hr postinjection compared to vehicle injection. Plasma pancreatic polypeptide concentration was increased by the high fat diet at 2 hr postfeeding (P < 0.02) but was lowered by MK-329 at 1 hr postinjection (P < 0.001). Plasma insulin was lowered by the high fat diet (P < 0.01) but was not affected by injections. The elevated plasma cholecystokinin concentration may have mediated depressed feed intake of dairy cattle fed the high fat diets.     

Radiographic contrast study of the upper gastrointestinal tract of eight dogs using carboxymethylcellulose mixed with a low concentration of barium sulphate

To evaluate the efficacy of a mixture of barium and carboxymethylcellulose (CMC) suspension in upper gastrointestinal contrast studies, and to determine its optimal concentration and method for use, eight healthy beagle dogs underwent conventional upper gastrointestinal contrast imaging and a modified procedure using a mixture of barium sulphate and CMC. Four different procedures were carried out on the dogs at weekly intervals. In the first, the dogs were administered 10 ml/kg of 60 per cent barium suspension for conventional study of the upper gastrointestinal tract. The second, third and fourth procedures involved modified methods in which the dogs were given 10 ml/kg of a mixture of 25 per cent barium and 0.5 per cent CMC prepared to a low, moderate or high viscosity, respectively. All four procedures were evaluated by four criteria: the quality of the images obtained, the translucency and distensibility of the bowel, and the transit time. The method of imaging which used a moderately viscious CMC formulation provided excellent image quality, high translucency, a rapid transit time and suitable bowel distension to enable more precise diagnosis of disorders of the gastrointestinal tract.