Phenotypic and genotypic characterization of canine pyoderma isolates of Staphylococcus pseudintermedius for biofilm formation

Biofilm-forming ability is increasingly being recognized as an important virulence factor in several Staphylococcus species. This study evaluated the biofilm-forming ability of sixty canine derived clinical isolates of S. pseudintermedius, using three phenotypic methods, microtiter plate test (MtP), Congo red agar method (CRA) and tube adherence test, and the presence and impact of biofilm-associated genes (icaA and icaD). The results showed that icaA and icaD genes were detected concomitantly in 55 (91.7%) of 60 isolates. A majority (88.3%) of the strains screened had matching results by the tube adherence test, MtP and PCR analysis. Better agreement (95%) was found between the PCR-based analysis and the CRA. Results of the icaA and icaD gene PCRs showed good agreement with CRA results, with a kappa of 0.7. Comparing the phenotypic methods, the statistical analysis showed that the agreement among the phenotypical tests using categorical data was generally good. Considering two classes (biofilm producer and biofilm non-producer), the percentage of matching results between the CRA method and the tube adherence test and between the CRA method and the MtP was 93.3%. A concordance of 100% was revealed between the MtP and the tube adherence test. The results indicate a high prevalence of the ica genes within S. pseudintermedius isolates, and their presence is associated with in vitro formation of a biofilm. A combination of phenotypic and genotypic tests is recommended for investigating biofilm formation in S. pseudintermedius.     

Treatment of tropical forages with exogenous fibrolytic enzymes: effects on chemical composition and in vitro rumen fermentation

The effects of three treatments of fibrolytic enzymes (cellulase from Trichoderma longibrachiatum (CEL), xylanase from rumen micro‐organisms (XYL) and a 1:1 mixture of CEL and XYL (MIX) on the in vitro fermentation of two samples of Pennisetum clandestinum (P1 and P2), two samples of Dichanthium aristatum (D1 and D2) and one sample of each Acacia decurrens and Acacia mangium (A1 and A2) were investigated. The first experiment compared the effects of two methods of applying the enzymes to forages, either at the time of incubation or 24 h before, on the in vitro gas production. In general, the 24 h pre‐treatment resulted in higher values of gas production rate, and this application method was chosen for a second study investigating the effects of enzymes on chemical composition and in vitro fermentation of forages. The pre‐treatment with CEL for 24 h reduced (p < 0.05) the content of neutral detergent fibre (NDF) of P1, P2, D1 and D2, and that of MIX reduced the NDF content of P1 and D1, but XYL had no effect on any forage. The CEL treatment increased (p < 0.05) total volatile fatty acid (VFA) production for all forages (ranging from 8.6% to 22.7%), but in general, no effects of MIX and XYL were observed. For both P. clandestinum samples, CEL treatment reduced (p < 0.05) the molar proportion of acetate and increased (p < 0.05) that of butyrate, but only subtle changes in VFA profile were observed for the rest of forages. Under the conditions of the present experiment, the treatment of tropical forages with CEL stimulated their in vitro ruminal fermentation, but XYL did not produce any positive effect. These results showed clearly that effectiveness of enzymes varied with the incubated forage and further study is warranted to investigate specific, optimal enzyme‐substrate combinations.     

Effect of Mediterranean saltbush (Atriplex halimus) ensilaging with two developed enzyme cocktails on feed intake,nutrient digestibility and ruminal fermentation in sheep

The aim of this study was to assess the effects of feeding Atriplex halimus (AH) silage treated with two developed enzyme cocktails to sheep on feed intake, nutrient digestibility and ruminal fermentation. The AH silage was treated without or with 2 L of ZAD1^® or ZAD2^®/1000 kg with 5% molasses and ensiled for 30 days. Barley grain (300 g/head/day) was fed as an energy supplement once daily at 10.00 hours and AH silage with or without enzyme treatment was offered ad libitum to animals twice daily at 09.00 and 16.00 hours. Sheep were fed on four experimental forage diets comprised of AH silage and barley (D1), AH silage treated with ZAD1^® and barley (D2), AH silage treated with ZAD2^® and barley (D3) and AH silage treated with a combination of ZAD1^® and ZAD2^® (1:1) and barley (D4). Ensiling AH with enzymes reduced its contents of neutral detergent fiber and acid detergent fiber. The dry matter intake of AH of D2, D3 and D4 decreased (P < 0.001) as compared to D1. However, enzyme‐treated diets had greater total digestible nutrients intake (P < 0.001) as compared to D1. The nutrients digestibility for D2, D3 and D4 were higher than those for D1 (P < 0.001), and were higher for D3 as compared to both D2 and D4. Sheep fed on D3 had highest (P < 0.001) ruminal total volatile fatty acids concentration, ammonia nitrogen concentration and microbial protein yield. It could be concluded that AH silage treated with ZAD1^® or ZAD2^® improved digestibility and rumen fermentation in sheep.     

Biology,incidence and host susceptibility of Pineus boerneri (Hemiptera: Adelgidae) in Colombian pine plantations

Adelgids (Hemiptera) in the genus Pineus have been reported as introduced insect pests causing serious losses to Pinus plantations worldwide. In 2008, Pineus boerneri was recorded for the first time in Colombia, with infestations noted on Pinus kesiya, P. tecunumanii, P. maximinoi and P. oocarpa. The lack of information on this insect in Colombia prompted investigations of its life cycle and infestation levels as well as host susceptibility of the main Pinus species planted in Colombia. In addition, the possibility of using a Ceraeochrysa species, an already established predator of adelgids in Colombia, for biological control was considered. Results showed that Pineus boerneri in Colombia has an anholocyclic life cycle comprised of four instars with a complete duration of between 49 and 97 d. Infestations were higher in the middle and upper part of trees. Pinus kesiya and P. maximinoi had the highest levels of susceptibility in field as well as in greenhouse trials. A survey of naturally infested trees showed P. tecunumanii to be moderately susceptible, whereas P. patula and P. oocarpa had low levels of susceptibility in a greenhouse trial but were not susceptible in the field. Investigations considering the impact of predation of Ceraeochrysa species showed a high predation rate of up to 140 P. boerneri consumed per day by a single Ceraeochrysa individual. Other predators of P. boerneri were recorded but were not sufficiently common to warrant detailed study.     

Physiological and metabolic properties of a digestion-resistant maltodextrin, classified as type 3 retrograded resistant starch

There is a growing interest in highly fermentable dietary fibers having the potential to reduce risks of disease through the production of short-chain fatty acids (SCFA). Recently a digestion-resistant retrograded maltodextrin (RRM), classified as type 3 resistant starch was developed. Systematic work to determine its molecular and physiological properties was carried out to determine (1) the fraction resistant to digestion in vitro and in vivo, (2) its postconsumption effect on blood glucose in healthy volunteers, and (3) its in vitro fermentation pattern, at different ages, by use of pooled fresh human fecal inoculum. RESULTS: The digestion resistant fraction obtained in vivo from ileostomy patients (59.4%) is similar to that obtained by the AOAC method for measuring retrograded resistant starch (59.7%). The relative glycemic response after consumption of 50 g of RRM was 58.5% compared to glucose set as 100%. When exposed to colonic microbiota, in vitro obtained indigestible fractions behave similarly to those obtained in vivo in ileostomy patients. Fermentation of RRM and production of butyric acid is negligible during the first months of life but develops subsequently during weaning. In adults, RRM fermentation results in a high yield of SCFA, with butyrate representing 21-31 mol % of total SCFA. The high yield of SCFA during colonic fermentation, observed from weaning age on, as well as the potential to help reduce glycemic load may be of benefit to a number of health-related functions in the host. Further study on clear clinical end points is warranted.     

Comparative study of health properties and nutritional value of durian, mangosteen, and snake fruit: experiments in vitro and in vivo

In vitro and in vivo studies of the health and nutritional properties of durian (Durio zibethinus Murr.) were compared with snake fruit (Salacca edulis Reinw.) and mangosteen (Garcinia mangostana). Dietary fibers, minerals, and trace metals were comparable. Total polyphenols (mg of GAE/100 g of FW) and flavonoids (85.1+/-6.1) were significantly higher (p<0.05) than in snake fruit (217.1+/-13.2 (mg of CE/100 g of FW)), durian (309.7+/-19.3 and 61.2+/-4.9), and mangosteen (190.3+/-12.1 and 54.1+/-3.8). Antioxidant activity (microM TE/100 g of FW) of durian measured by DPPH and ABTS assays (228.2+/-13.4 and 2016.3+/-81.1) was significantly higher (p<0.05) than in snake fruit (110.4+/-7.9 and 1507.5+/-70.1) and mangosteen (79.1+/-5.9 and 1268.6+/-62.3). HPLC/DAD analysis of durian (microg/100 g of FW) showed that quercetin (1214.23+/-116.7) was present at levels three times that of caffeic acid, and twice as high as p-coumaric and cinnamic acids. The correlation coefficients between the bioactive compounds of fruits and their antioxidant activities were high (R2=0.99). Male Wistar rats (25) were divided into five dietary groups: the control group was fed the basal diet (BD); in addition to BD, the cholesterol (Chol) group was supplemented with 1% of Chol; the diets of the Chol/Durian, Chol/Snake, and Chol/Mangosteen groups were supplemanted with 5% of these fruits, respectively. It was found that diets supplemented with durian, and to a lesser degree with snake fruit and mangosteen, significantly hindered the rise in plasma lipids and the decrease in antioxidant activity. The nutritional values were comparably high. In conclusion, it could be suggested that inclusion of studied tropical fruits, especially durian, in known disease-preventing diets could be beneficial.     

Antioxidant profile of red wines evaluated by total antioxidant capacity, scavenger activity, and biomarkers of oxidative stress methodologies

The study of the antioxidant capacity of foodstuffs requires the use of diverse determination methods to gain a wider picture of their multiple effects. The aim of this work was to evaluate the "antioxidant profile" of red wines applying TAC (total antioxidant capacity) methods: 2,2'-azinobis(3-ethyl-benzthiazoline-6-sulfonic acid) (ABTS), 2,2-diphenyl-1-picrylhydrazyl, N,N-dimethyl-p-phenylenediamine dihydrochloride, oxygen radical absorbance capacity, ferric reducing/antioxidant power, hydroxyl and superoxide radical scavenger activities, and biomarkers of oxidative stress methods such as lipid peroxidation inhibition and inhibition of damage to DNA. Furthermore, levels of total polyphenols (TPP) of wines were also evaluated. Three bottles of 107 different Spanish red wines (total samples 321), made from different grape varieties, aging processes, and vintages, were analyzed. The validation of TAC methods, the first step in this work, provided a good linearity, proportionality, and low detection limits. Among these methods, the ABTS was the most satisfactory for its rapidity, cost, and precision. All wines showed an important capacity to scavenge hydroxyl radicals and were capable of blocking superoxide radicals but with 10 times lower intensity. Wines also showed important protective action on biomarkers of oxidative stress; they were much more active to inhibit lipid peroxidation than DNA oxidation. Few statistically significant correlations among levels of TPP and antioxidant properties of wines were detected. Furthermore, values of these correlations were very low.     

Method of determination of nitrosamines in sausages by CO2 supercritical fluid extraction (SFE) and micellar electrokinetic chromatography (MEKC)

In this paper, the use of supercritical fluid extraction (SFE) and micellar electrokinetic capillary chromatography (MEKC) is proposed for the complete analysis of volatile nitrosamines in sausages. The extraction fluid used was CO2 and variables such as density, temperature of thimbles, extraction time, modifier, fluid flow, and kind of traps were investigated. Several experiments were carried out to obtain the most favorable conditions for analysis of volatile nitrosamines in sausages. The recoveries ranged from 21 to 82% for the five nitrosamines studied. The optimal condition of extraction was 0.2 g of sample fortified with 10 mg/kg, using dynamic extraction during 20 min and with adsorbent Florisil in the trap. The solvent selected for the elution of the analytes was methanol.