Phytosome-conjugated carvacrol: A novel approach for improving growth performance, intestinal morphology and economics of production in Broiler Chicken

Essential oils are plant-derived aromatic volatile oils, and they contain bioactive compounds that have been shown to improve poultry nutrition. However, considering problems associated with the solubility and bioavailability of polyphenolic compounds, the study was planned to find out the effect of the novel feed-grade delivery system, phytosomes for conjugation of plant-derived polyphenolic compound carvacrol on the growth performance of broiler chickens. The experiment was conducted, on 240 broiler chicks for a period of 6 weeks. The chicks were divided into 4 groups having 4 replicates of 15 birds each. The birds in the control group (T0) offered a standard diet as per BIS (2007) specification. Group T1 received a standard diet supplemented with Bacitracin Methylene Disalicylate (BMD) antibiotic at standard dose and group T2 received a standard diet supplemented with carvacrol essential oil @100 mg/kg feed. Group T3 received a standard diet supplemented with phytosome-conjugated carvacrol essential oil (carvacrol @16.6%) @100 mg/kg feed. The performance of all the treatment groups was assessed with respect to the different performance parameters. The supplementation of phytosome-conjugated carvacrol essential oil (carvacrol @16.6%) @ 100 mg/kg feed was found beneficial in terms of growth performance, feed efficiency, and intestinal morphometry. In terms of economics of broiler production, the results revealed that the addition of phytosome- conjugated carvacrol essential oil and carvacrol essential oil in diets was found beneficial in reducing the cost of broiler production, thereby enhancing the margin of profit in broiler production and fetching higher net profit than the control group

Removal of sub-micron size and dilute turbidity from service water by application of an electrochemical carbon filter

37-47A semi industrial scale graphite felt electrochemical filter (ECF) assembly has been used to remove iron and silica turbidity from domestic service water of Kalpakkam. Two test runs are performed. In the first run 17.21 m3 of water is processed and 2.15 × 104 NTU-liter turbidity is removed [inlet turbidity 1.67 NTU (nephelometric turbidity units), outlet turbidity 0.42 NTU, and efficiency 75%]. In the second run 6.67 m3 of water is processed and 3.87 × 103 NTU-liter turbidity is removed (inlet turbidity 0.85 NTU, outlet turbidity 0.27 NTU, and efficiency 68%). In this process non-reactive silica removal efficiency of 50% is observed. Nearly complete removal of E.coli is also shown by ECF. It is observed that 600-1200 l/h flow rate and 5.0-8.0 V applied potential with 1 ampere current are the optimized parameters for ECF operation. Teflon is identified as non swelling material in water for the carbon felt electrode isolation spacers and support plates. As ECF is exclusively used for submicron filtration, a macro filter prior to it is must. The ECF has the advantages of treating turbidity irrespective of the sign and magnitude of charge on the turbidity particles as the applied potential can be maneuvered accordingly

The Kallikrein-Kinin System: A Novel Mediator of IL-17-Driven Anti-<i>Candida</i> Immunity in the Kidney

<div><p>The incidence of life-threatening disseminated <i>Candida albicans</i> infections is increasing in hospitalized patients, with fatalities as high as 60%. Death from disseminated candidiasis in a significant percentage of cases is due to fungal invasion of the kidney, leading to renal failure. Treatment of candidiasis is hampered by drug toxicity, the emergence of antifungal drug resistance and lack of vaccines against fungal pathogens. IL-17 is a key mediator of defense against candidiasis. The underlying mechanisms of IL-17-mediated renal immunity have so far been assumed to occur solely through the regulation of antimicrobial mechanisms, particularly activation of neutrophils. Here, we identify an unexpected role for IL-17 in inducing the Kallikrein (Klk)-Kinin System (KKS) in <i>C</i>. <i>albicans</i>-infected kidney, and we show that the KKS provides significant renal protection in candidiasis. Microarray data indicated that Klk1 was upregulated in infected kidney in an IL-17-dependent manner. Overexpression of Klk1 or treatment with bradykinin rescued IL-17RA^-/- mice from candidiasis. Therapeutic manipulation of IL-17-KKS pathways restored renal function and prolonged survival by preventing apoptosis of renal cells following <i>C</i>. <i>albicans</i> infection. Furthermore, combining a minimally effective dose of fluconazole with bradykinin markedly improved survival compared to either drug alone. These results indicate that IL-17 not only limits fungal growth in the kidney, but also prevents renal tissue damage and preserves kidney function during disseminated candidiasis through the KKS. Since drugs targeting the KKS are approved clinically, these findings offer potential avenues for the treatment of this fatal nosocomial infection.</p></div

Combination of fluconazole and bradykinin increased survival of <i>C. albicans</i> infected mice.

<p><b>(A)</b> Experimental design for combinatorial fluconazole and bradykinin therapy strategy <b>(B)</b> WT mice (n = 16) were infected with <i>C</i>. <i>albicans</i>. Infected mice were treated with two doses of FLC only (5 mg/kg at 2 and 24 h p.i.), bradykinin only (300nmol/kg starting 1 day prior to infection and daily for 14 days) or a combination of FLC and bradykinin. Sham-infected mice (n = 3) were treated with FLC, bradykinin or both. Survival was assessed over 14 d. Data are combined from 3 independent experiments. p<0.05 (*), p>0.001 (***), p>0.0001 (****).</p

Bradykinin alleviated kidney injury and preserved renal function in <i>C. albicans</i> infected mice.

<p>WT mice (n = 14–20) were either left treated ± bradykinin (300 nmol/kg/day) starting day -1 (relative to infection). Mice were infected with <i>C</i>. <i>albicans</i> strains <b>(A)</b> (CAF2-1) or <b>(B)</b> (SC5314). Sham-infected WT mice were treated ± bradykinin (n = 3–5). Survival was assessed over 14 d. Data are pooled from <b>(A)</b> 4 and <b>(B)</b> 2 independent experiments. At day 7 p.i., mice were evaluated for <b>(C)</b> angioedema development in the hind paw (n = 4–7) and <b>(D)</b> serum BUN and creatinine levels (n = 6–10). Data pooled from 2–3 independent experiments. Each dot represents one mouse and the bars indicate mean. At day 7 p.i., kidney sections were evaluated for <b>(E)</b> histopathology and inflammatory cell influx by PAS staining (n = 8) and <b>(F)</b> NGAL expression by IHC (n = 6–8). Black arrows indicate tubular damage and atropy; * indicates inflammatory cell influx. Representative photomicrographs from 2 independent experiments. Original magnification: 100X. <b>(G)</b> Cell lysates of kidney homogenates (n = 5–6) were evaluated for NGAL by western blotting. Images were enumerated using ImageJ. Representative image of 1 of 2 independent experiments. Bars indicate mean ± S.D and combined from 2 independent experiments. P <0.05 (*), <0.01 (**). <0.001 (***), <0.0001 (****). ns, not significant.</p

The expression of Klk is impaired in IL-17RA^-/- kidney following <i>C. albicans</i> infection.

<p><b>(A)</b> WT and IL-17RA^-/- mice (n = 4–6) were subjected to systemic <i>C</i>. <i>albicans</i> (CAF2-1 or SC5314) infection. After 48 h, kidneys were evaluated for fungal load. Data pooled from 2–3 independent experiments. <b>(B)</b> Heat map representing averaged intensity of expression of genes in WT and IL-17RA^-/- kidneys (n = 2) at 48 h p.i. <b>(C)</b> Schematic representation of Kallikrein-kinin system (KKS). (<b>D)</b> Kidneys of WT and IL-17RA^-/- mice (n = 6) were evaluated for expression of <i>Klk1</i> and <i>Klk1b27</i> at 48 h p.i. Data pooled from 2 independent experiments. At 72 h p.i., whole cell extracts from WT and IL-17RA^-/- kidneys (n = 5–6) infected with <i>C</i>. <i>albicans</i> <b>(E)</b> CAF2-1 or <b>(F)</b> SC5314 were evaluated for Klk1 protein by western blotting. Sham infected mice received PBS. Images were quantified using ImageJ. Representative image of 1 of 2 independent experiments (<b>E</b> and <b>F</b>). For the bar diagram, data are combined from 2 independent experiments. Bars indicate mean ± S.D. <i>P</i><0.05 (*), <0.01 (**), <0.001 (***). ns, not significant.</p

Minimal impact of bradykinin treatment on fungal clearance and inflammatory cells influx in the <i>C. albicans</i> infected kidney.

<p>WT mice were treated ± bradykinin (300 nmol/kg/day) or PBS on day -1 (relative to infection). <b>(A)</b> Kidneys were evaluated for fungal load on days 3 (n = 3–4) and 7 (n = 8–11) p.i. (<b>B</b>) At day 7 p.i. (n = 4–6), kidney infiltrating neutrophils (Gr1⁺) and macrophages (F4/80⁺) (gated on CD45⁺) cells were evaluated by flow cytometry. Data are combined from 2 independent experiments for <b>(A)</b> and <b>(B)</b>. Each dot represents one mouse, and the bars indicate mean. <b>(C)</b> BM-derived neutrophils and BMDMs from WT mice were incubated <i>in vitro</i> with unopsonized <i>C</i>. <i>albicans</i> yeast ± bradykinin for 3 h, and fungal load was determined by plating culture supernatants. Percentage of <i>C</i>. <i>albicans</i> killed by neutrophils and macrophages is shown. <b>(D)</b> IL-6 in RTEC conditioned media was assessed after 24 h bradykinin treatment. <b>(E)</b> IL-6 and TNFα and <b>(F)</b> nitrite levels in the supernatants of BMDMs treated ± bradykinin or LPS (1 ng/ml) for 24 h. Data are representative of 3 independent experiments for <b>(C-F)</b>. Bars indicate mean ± S.D. p <0.01 (**). ns, not significant.</p