[A multicenter prospective study on early identification of refractory Mycoplasma pneumoniae pneumonia in children].

Objective: To explore potential predictors of refractory Mycoplasma pneumoniae pneumonia (RMPP) in early stage. Methods: The prospective multicenter study was conducted in Zhejiang, China from May 1st, 2019 to January 31st, 2020. A total of 1 428 patients with fever >48 hours to <120 hours were studied. Their clinical data and oral pharyngeal swab samples were collected; Mycoplasma pneumoniae DNA in pharyngeal swab specimens was detected. Patients with positive Mycoplasma pneumoniae DNA results underwent a series of tests, including chest X-ray, complete blood count, C-reactive protein, lactate dehydrogenase (LDH), and procalcitonin. According to the occurrence of RMPP, the patients were divided into two groups, RMPP group and general Mycoplasma pneumoniae pneumonia (GMPP) group. Measurement data between the 2 groups were compared using Mann-Whitney U test. Logistic regression analyses were used to examine the associations between clinical data and RMPP. Receiver operating characteristic (ROC) curves were used to analyse the power of the markers for predicting RMPP. Results: A total of 1 428 patients finished the study, with 801 boys and 627 girls, aged 4.3 (2.7, 6.3) years. Mycoplasma pneumoniae DNA was positive in 534 cases (37.4%), of whom 446 cases (83.5%) were diagnosed with Mycoplasma pneumoniae pneumonia, including 251 boys and 195 girls, aged 5.2 (3.3, 6.9) years. Macrolides-resistant variation was positive in 410 cases (91.9%). Fifty-five cases were with RMPP, 391 cases with GMPP. The peak body temperature before the first visit and LDH levels in RMPP patients were higher than that in GMPP patients (39.6 (39.1, 40.0) vs. 39.2 (38.9, 39.7) ℃, 333 (279, 392) vs. 311 (259, 359) U/L, both P<0.05). Logistic regression showed the prediction probability π=exp (-29.7+0.667×Peak body temperature (℃)+0.004×LDH (U/L))/(1+exp (-29.7+0.667×Peak body temperature (℃)+0.004 × LDH (U/L))), the cut-off value to predict RMPP was 0.12, with a consensus of probability forecast of 0.89, sensitivity of 0.89, and specificity of 0.67; and the area under ROC curve was 0.682 (95%CI 0.593-0.771, P<0.01). Conclusion: In MPP patients with fever over 48 to <120 hours, a prediction probability π of RMPP can be calculated based on the peak body temperature and LDH level before the first visit, which can facilitate early identification of RMPP.

Dietary choline supplementation regulated lipid profiles of egg yolk, blood, and liver and improved hepatic redox status in laying hens.

Five hundred and forty 19-wk-old HyLine Brown laying hens were randomly distributed to 6 dietary treatments and fed 1of 6 corn-soybean meal-based diets added into choline with 0, 425, 850, 1,700, 3,400, and 6,800 mg/kg to investigate effects of dietary choline supplementation on lipid profiles of egg yolk, serum and liver, and hepatic redox status of laying hens. Yolk weight and total lipid, triglyceride, cholesterol and phosphatidylcholine, serum triglyceride, cholesterol, apolipoprotein B 100 (apoB 100), and very low density lipoprotein (VLDL), and liver relative weight, total lipid, triglyceride and apoB 100 as well as hepatic total superoxide dismutase and glutathione peroxidase (GSH-Px) activities, total antioxidant capacity (T-AOC), and malondialdehyde (MDA) in laying hens at weeks 58 and 68 of age were determined. The differences (P < 0.001) were caused by choline treatments in yolk phosphatidylcholine (at 850 mg/kg or more choline), serum VLDL, and liver triglyceride (at 1,700 and 3,400 mg/kg choline) of birds, at weeks 58 and 68 of age, and yolk total lipids were elevated (P < 0.05) by supplemental choline at 3,400 mg/kg whereas liver total lipids were reduced (P < 0.05) by 1,700 and 3,400 mg/kg choline addition. Hens fed diets supplemented choline had higher (P = 0.005) liver GSH-Px activity (with 3,400 mg/kg choline) and greater (P = 0.014) T-AOC (with 1,700 mg/kg choline) than those fed diets with 0 and 425 mg/kg choline addition. Choline affected serum VLDL, liver total lipid, triglyceride and apoB 100 at weeks 58 and 68 of age and hepatic GSH-Px activity, T-AOC and MDA at week 68 of age quadratically (P < 0.05), whereas it influenced total lipid and phosphatidylcholine of egg yolk linearly (P < 0.05) and quadratically (P < 0.05). In conclusion, dietary choline supplementation elevated yolk total lipid and phosphatidylcholine and serum VLDL, reduced liver total lipid and triglyceride, and enhanced hepatic GSH-Px activity and T-AOC in laying hens.

High-throughput sequencing reveals the effect of Bacillus subtilis CGMCC 1.921 on the cecal microbiota and gene expression in ileum mucosa of laying hens.

This study evaluated the effects of Bacillus subtilis CGMCC 1.921 supplementation on the production performance, cecal microbiota and mucosal transcriptome of laying hens by 16s rRNA gene sequencing and RNA-seq. A total of 144 27-week-old Hy-Line Brown laying hens were allocated into two treatments, namely, a basal diet without additions (T0) and the basal diet supplemented with 1.0 × 108 cfu/g (T1) B. subtilis CGMCC 1.921, with six replicates of 12 birds in each for 24 weeks. The results showed that T1 significantly decreased feed:egg ratio compared with T0 (P < 0.05). Dietary supplementation with B. subtilis CGMCC 1.921 increased the Shannon index (P < 0.05) which indicated enhanced diversity of cecal microflora. An increasing trend in Observed species index (P = 0.072) was observed in hens fed with diets supplemented with B. subtilis CGMCC 1.921 that showed a higher species richness. And T1 modulated cecal microbiota by increasing the relative proportion of Alistipes, Subdoligranulum, Ruminococcaceae UCG-014, Anaerotruncus, Ruminiclostridium 5, Ruminococcaceae UCG-010, Erysipelatoclostridium, Ruminococcaceae UCG-009, Family XIII AD3011 group, Bacillus, Faecalicoccus, Firmicutes bacterium CAG822, Oxalobacter, and Dielma at genus level (P < 0.05). In addition, there was a tendency of increase in the relative abundance of Lactobacillus (P = 0.055), Anaerobiospirillum (P = 0.059) and Family XIII UCG-001 (P = 0.054), Peptococcus (P = 0.078), and Ruminococcaceae UCG-004 (P = 0.078). Moreover, heatmap analysis indicated that the abundance of Campylobacter and Clostridium sensu stricto 1 was lower than T0. A total of 942 genes were identified by differential expression analysis, among which 400 genes were upregulated and 542 genes were downregulated. Bioinformatics analysis suggested that the upregulated genes were involved in Peroxisome Proliferator Activated Receptor (PPAR) signaling pathway, starch and sucrose metabolism, glycine/serine/threonine metabolism, and galactose metabolism, which may promote nutrient absorption. This study provided novel insights into the probiotic mechanisms of B. subtilis on laying hens.

Comparative Effect of Dietary Soybean Oil, Fish Oil, and Coconut Oil on Performance, Egg Quality and Some Blood Parameters in Laying Hens.

Two hundred and sixteen 28-wk-old Hy-line laying hens were randomly distributed to three dietary treatments and fed 1of 3 diets containing 8% soybean oil, fish oil, or coconut oil from 28 to 47 wk of age to investigate comparative effect of dietary soybean oil, fish oil, and coconut oil on the performance, egg quality and blood malondialdehyde (MDA), aspartate transaminase (AST) and uric acid (UA). Hens fed fish oil showed poor performance compared with soybean oil or coconut oil, and especially egg weight throughout the trial was significantly and consistently decreased (P < 0.05) due to dietary fish oil. Unexpectedly, shell reflectivity throughout the majority of the trial was consistently and significantly higher (P < 0.05) when hens fed fish oil than that when fed soybean oil or coconut oil. Dietary treatments affected (P < 0.05) shell shape at 4 of 8 time points tested. Average shell shape in fish oil treatment was higher (P < 0.05) than that of coconut oil group. Albumen height, Haugh unit and yolk color were influenced by dietary treatments only at 1 or 2 time points. However, average albumen height and Haugh unit in fish oil treatment were higher (P < 0.05) than that of soybean oil or coconut oil treatments and average yolk color in coconut oil treatment was higher (P < 0.05) than that of soybean oil group. Serum MDA, AST and UA concentrations were increased (P < 0.05) by fish oil during the majority of the first 2 mo of the trial. These data suggested that the inclusion of fish oil into feed may reduce the performance of laying hens, especially the egg weight, decrease the intensity of egg brown color and increase blood MDA, AST and UA levels compared with soybean oil or coconut oil. As a result, hens fed fish oil may lay smaller, longer and lighter-brown eggs whereas those fed coconut oil produce blunter and darker-brown eggs relative to soybean oil.

Effects of long-term Bacillus subtilis CGMCC 1.921 supplementation on performance, egg quality, and fecal and cecal microbiota of laying hens.

This study evaluated the effects of long-term Bacillus subtilis CGMCC 1.921 supplementation on the performance, egg quality, and fecal/cecal microbiota of laying hens. A total of 360 28-week-old Hy-Line Brown laying hens were randomly allocated into 5 treatments with 6 replicates of 12 birds each for 24 weeks. The experimental treatments included a basal diet without additions (Con) and the basal diet supplemented with 1.0 × 105 (B1), 1.0 × 106 (B2), 1.0 × 107 (B3), and 1.0 × 108 (B4) cfu/g B. subtilis CGMCC 1.921. The results showed that feed:egg ratio significantly decreased (P < 0.05) in groups B1 (wk 13 to 16, 17 to 20, 21 to 24, and one to 24), B2 (wk 13 to 16, 17 to 20, and 21 to 24), B3 (wk 13 to 16, 17 to 20, 21 to 24, and one to 24), and B4 (wk 13 to 16, 17 to 20, 21 to 24, and one to 24). However, egg production, egg weight, and feed intake were not significantly different (P > 0.05) among treatments. Eggshell strength significantly improved (P < 0.05) in groups B1 (wk 8, 16, 20, and 24), B2 (wk 20 and 24), and B3 (wk 8, 16, 20, and 24). Fecal E. coli counts significantly decreased (P < 0.05) in groups B1 (wk 16), B2 (wk 12, 16, 20, and 24), B3 (wk 12, 20, and 24), and B4 (wk 16, 20, and 24). Lactobacillus in cecal digesta of groups B1, B3, and B4 increased significantly (P < 0.01). Bifidobacterium in cecal digesta of groups B1, B2, B3, and B4 increased significantly (P < 0.05). Bifidobacterium counts increased linearly (P = 0.015) and quadratically (P = 0.004) as B. subtilis CGMCC 1.921 supplementation increased. Compared with Con, E. coli in the cecal digesta of groups B2 and B4 decreased significantly (P < 0.01). C. perfringens in the cecal digesta of groups B3 and B4 decreased significantly (P < 0.05). E. coli:Lactobacillus ratio decreased in group B1 (P < 0.05) and B2, B3, and B4 (P < 0.01). Therefore, the probiotic B. subtilis CGMCC 1.921 effectively improved performance and egg quality via the reduction of fecal E. coli and beneficial modulation of cecal microbiota.

Comparative study of DL-selenomethionine vs sodium selenite and seleno-yeast on antioxidant activity and selenium status in laying hens.

The aim of this study was to compare the effect of DL-selenomethionine (SM) with 2 routinely used Se sources, sodium selenite (SS) and seleno-yeast (SY), on relative bioavailability based on antioxidant activity and tissue Se content. Six hundred thirty 131-day-old brown laying hens were randomly assigned to 7 treatments for 168 d (24 wks) with 6 replicates of 15 hens per replicate. The SS and SY animals were supplemented a cornmeal and soybean diet that supplied a total Se 0.3 mg/kg whereas SM was added at 4 different levels to the total Se at 0.1, 0.3, 0.5 and 0.7 mg/kg. All hens fed the Se-supplemented diet showed higher glutathione peroxidase (GSH-Px) activity (P < 0.01), higher superoxide dismutase (SOD) activity (P < 0.05), lower malondialdehyde (MDA) content (P < 0.05) in plasma, and greater Se contents in egg yolks, albumen, leg muscle, breast muscle, liver, and plasma compared with those fed the control diet (P < 0.01). The organic sources (SY and SM) exhibited a greater ability to increase the GSH-Px activity (P < 0.01) and Se content in albumen (P < 0.01), leg, and breast muscles (P = 0.0099 and P = 0.0014, respectively) than the SS that was added at 0.3 mg Se/kg. The higher SM added levels increased the GSH-Px activity until the dose of 0.5mg Se/kg (P < 0.01).The greater Se concentrations in albumen, muscle and liver appeared in the higher SM-added level, as well as above the dose of 0.1 mg Se/kg (P < 0.01). In addition, hens fed the diet with SM accumulated more Se in albumen, leg, and breast muscle than those fed diets with SY (P < 0.05). These results confirmed the higher ability of organic Se sources to increase the antioxidant activity and Se deposition in egg albumen, leg, and breast muscles compared with SS, and demonstrated a significantly better efficiency of SM compared with SY for albumen and muscle Se enrichment.

Effect of antagonistic yeast XL-1 on resistance-associated enzyme activities in postharvest cantaloupe.

The effect of the antagonistic yeast XL-1 on resistance-associated enzyme activities in postharvest cantaloupe was studied by inoculating the antagonistic yeast XL-1. Cantaloupes were sterilized, dried in air, and soaked in antagonistic yeast treatment liquid for 30 s. After drying in air, the cantaloupe was stored at room temperature (2°-5°C). The activities of resistance-associated enzymes in cantaloupe like polyphenol oxidase, ß-1,3-glucanase, peroxidase, and superoxide dismutase were measured every 7 days. Our results indicated that the antagonistic yeast XL-1 significantly improved the activity of ß-1,3-glucanase and chitinase to promote the disease resistance of postharvest cantaloupe.

Long-term effects of choline on productive performance and egg quality of brown-egg laying hens.

A total of five hundred forty 19-wk-old HyLine Brown hens were used to study the long-term effects of increasing choline with 0 (control), 425, 850, 1,700, 3,400, and 6,800 mg/kg of corn-soybean meal-based diets on productive performance and egg quality. Phase 1 was from 19 to 58 wk, and phase 2 was from 59 to 68 wk. During the whole experimental period, dietary choline had no significant effects on feed intake, egg weight, and egg mass (P > 0.05). During phase 1, egg production decreased linearly (P < 0.05) and feed conversion ratio (FCR) tended to increase linearly (P = 0.057) with increasing choline level in the diet. Moreover, BW decreased both linearly (P < 0.01) and quadratically (P < 0.05) as choline increased from 0 to 6,800 mg/kg. No significant treatment effects were found for shell thickness and shell strength of eggs (P > 0.05). However, albumen height and Haugh units increased linearly (P < 0.01 and P < 0.05, respectively) as choline increased during phase 2. Compared with the control group, diets supplemented with 425 or 850 mg of choline/kg significantly (P < 0.01) improved yolk color during phase 1. This study indicates that a dietary choline level of no more than 700 mg/kg is sufficient to maintain egg production. The effect of choline on egg quality was minimal when hens were fed a corn-soybean meal-based diet from 19 to 68 wk of age.

Outbreak of microsporidial keratoconjunctivitis with rugby sport due to soil exposure.

AIMS: To investigate a cluster of microsporidial keratoconjunctivitis in 33 eyes of 25 previously healthy paediatric and teenage individuals after a rugby match. METHODS: An observational case series was reported. Analysis of medical record of patients with microsporidial keratoconjunctivitis, who presented within May 2012, was performed. All patients were treated by a single ophthalmologist with a standardized topical regime, including a fluoroquinolone (moxifloxacin) and an antiseptic (Brolene or Desomedine). Five eyes received corneal scrapings. RESULTS: The mean age was 13.36 years (range 5-16). All patients have participated in a rugby match on 21-22 April 2012. The onset of symptoms ranged from 10 to 30 days post exposure. All eyes had multiple superficial coarse punctate keratitis. Four (12%) eyes presented with keratic precipitates. One (3%) eye had intraocular pressure of 27 mm Hg. Microscopic examination of corneal scrapings with modified trichrome or calcofluor white (CFW) fluorescent staining was unremarkable but subsequent PCR test was positive for the small subunit rRNA gene of Vittaforma corneae in three out of five eyes. Sequencing of the PCR product of 1150 bp showed 96-100% identity with the Indian or Singaporean strains of V. corneae. After treatment, all eyes healed without sequel. CONCLUSIONS: The first outbreak of microsporidial keratoconjunctivitis in paediatric and teenage individuals with a rugby match is reported. A standardized topical regime, including a fluoroquinolone (moxifloxacin) and an antiseptic (Brolene or Desomedine), seems to be safe and effective, and requires validation in future treatment trials.

High levels of vitamin E affect retinol binding protein but not CYP26A1 in liver and hepatocytes from laying hens.

This study was conducted to determine the influence of vitamin E on the retinol binding protein (RBP) and cytochrome p450 family 26 subfamily A polypeptide 1 (CYP26A1), which are specific transporters and catabolic enzymes of vitamin A, respectively. In the in vivo experiments, a total of 450 laying hens was fed 5 levels of vitamin E (0, 20, 80, 320, and 1,280 IU/kg of feed) supplementation. For the in vitro assays, hepatocytes from laying hens were cultured in 4 levels of α-tocopherol (0, 10, 50, and 100 µM). High dietary vitamin E increased the concentration of vitamin A in liver (P < 0.05). The RBP and its mRNA expression in liver and hepatocytes were markedly inhibited by dietary vitamin E (320 and 1,280 IU/kg) and α-tocopherol (100 µM) in culture medium (P < 0.05). However, CYP26A1 and its mRNA expression were not affected by vitamin E in both liver and hepatocytes (P > 0.05). The results indicate that excessive vitamin E could increase the concentration of vitamin A in liver by inhibiting RBP synthesis in hepatocytes.

The probiotic Bacillus licheniformis ameliorates heat stress-induced impairment of egg production, gut morphology, and intestinal mucosal immunity in laying hens.

We investigated the effect of a 12-d exposure to 34°C plus dietary inclusion of the probiotic Bacillus licheniformis on the egg production, gut morphology, and intestinal mucosal immunity of laying hens. Ninety-six commercial hens (Hy-Line Brown) at the age of 60 wk were randomly allocated to 4 groups. After a period of laying rate adjustment (14 d), all the hens were subjected to 2 temperature treatments (12 d). Birds in 1 group were raised at 21°C and fed a basal diet, and birds in the other 3 groups were raised at 34°C and fed a basal diet supplemented with 0, 10(6), or 10(7) cfu of B. licheniformis per gram of feed, respectively. Rearing at 34°C depressed egg production and feed intake (P < 0.05). Compared with birds kept at 21°C, birds kept at 34°C had elevated serum levels of tumor necrosis factor-α (d 6), IL-1 (d 6 and 12), and corticosterone (d 6); decreased villus height (ileum: d 6; cecum: d 6 and 12) and ratio of villus height to crypt depth (ileum: d 6; cecum: d 6 and 12); fewer intraepithelial lymphocytes (ileum: d 6; cecum: d 6) and IgA-secreting cells (ileum: d 6; cecum: d 6 and 12); and more mast cells (ileum: d 6; cecum: d 6 and 12; P < 0.05). The number of goblet cells in the cecum increased at d 6 in heat-treated birds, and then deceased at d 12 (P < 0.05). Moreover, morphological examination showed injury to the villi of birds kept at 34°C. In general, inclusion of 10(7) cfu/g of B. licheniformis in the diet of heat-stressed hens was effective in overcoming the observed decline in egg production and feed intake, restoring the impaired villus structure, and sustaining a balanced mucosal immune response. Therefore, the probiotic B. licheniformis may be useful for ameliorating the adverse influence of heat on the egg production and gut health of laying hens.

Effects of an aqueous alfalfa extract on production performance, egg quality and lipid metabolism of laying hens.

The present study was conducted to evaluate the effect of an aqueous alfalfa extract (AAE) on production performance, egg quality and lipid metabolism of laying hens between 28 and 36 weeks of age. Four groups of commercial hens (Hy-Line Brown) were fed with diets containing 0, 0.05%, 0.1% and 0.15% AAE. Dietary AAE had no effect on egg production parameters (p > 0.05). Shell strength was increased (linear and quadratic effects; p < 0.05) with gradient addition of AAE. Furthermore, shell strength was notably higher (p < 0.05) in 0.1% and 0.15% AAE groups than the control group. With an exception of serum low and high density lipoprotein cholesterol, serum total triglycerides and liver cholesterol concentration (day 42), lipid indices in yolk, serum and liver were lowered (linear and quadratic effects; p < 0.05) at day 42 and day 56 as AAE in diets increased, as well as the enzyme activities of acetyl-CoA carboxylase (ACC) and ß-hydroxy-ß-methylglutaryl-CoA (HMG-CoA) reductase. In relation to the control group, cholesterol and triglycerides in yolk, liver and serum and serum very low density lipoprotein cholesterol were lower in 0.15% AAE-treated birds at day 42 and day 56. It also significantly decreased activities of ACC, HMG-CoA reductase and fatty acid synthetase (FAS) at day 56. The 0.1% AAE decreased triglycerides in yolk, liver and serum, as well as the activities of ACC and FAS at day 56. These data suggest that dietary AAE could be applied to improve egg shell quality and modify the lipid status of laying hens and eggs.

Effects of dietary polysavone (Alfalfa extract) and chlortetracycline supplementation on antioxidation and meat quality in broiler chickens.

1. A total of 360 1-d-old male commercial Arbor Acre broiler chickens were randomly assigned to 5 groups (6 replicates of 12 birds each) to evaluate the dietary effects of polysavone (0·5, 1·0 and 1·5 g/kg), a natural extract from alfalfa, and 0·15 g/kg chlortetracycline (CTC) on growth performance, antioxidation and meat quality of broiler chickens. 2. Over the 6-week study, feed intake increased significantly with CTC supplementation and final body weight (BW) was significantly higher for 1·0 g/kg polysavone and 0·15 g/kg CTC treatments. Feed:gain ratio was not significantly affected by the dietary treatments. 3. At 3 weeks of age, serum total superoxide dismutase (T-SOD) activity for all polysavone treatments was significantly higher than controls, liver T-SOD activity in 1·5 g/kg polysavone group was significantly higher than the control and CTC groups, and serum glutathione peroxidase (GSHPx) activity for 1·5 g/kg polysavone and liver GSHPx activity in all polysavone groups were significantly higher than CTC. 4. At 6 weeks of age, serum and liver T-SOD activity in 1·5 g/kg polysavone group and liver GSHPx activity for all polysavone treatments were higher significantly than the control and CTC groups, and serum malondialdehyde (MDA) content for all polysavone treatments was significantly lower than CTC. 5. Breast muscle T-SOD activity and pH value at 6 weeks of age were significantly higher and MDA content was significantly lower in 1·0 and 1·5 g/kg polysavone groups than in the control and CTC groups. Breast muscle shear force was significantly lower in l·5 g/kg polysavone group compared with the control, and drip loss for all polysavone treatments was significantly lower than CTC. 6. It was indicated that polysavone modulates antioxidation and modifies meat quality, but with no adverse effect on performance of broiler chickens, and that CTC can be beneficial to performance but has no beneficial effect on antioxidant function or meat quality.

Carry-over of melamine from feed to eggs and body tissues of laying hens.

The objective of this study was to assess the carry-over of melamine from feed into eggs and body tissues of laying hens. In the first experiment, laying hens were supplied with feed added at 0, 1, 2, 5, 25, 50, and 100 mg kg(-1) of melamine for 21 days followed by a depletion period to observe the residues of melamine in eggs. In a second experiment, laying hens were allocated 0, 50, and 100 mg kg(-1) melamine to determine levels of melamine in body tissues. Melamine and cyanuric acid were simultaneously analysed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) in the diet as well as in eggs and body tissue. In the first experiment, melamine appeared in the egg within 24 h after first ingestion of the melamine at 5, 25, 50, and 100 mg kg(-1). Melamine concentration in egg reached a maximum of 2.34 mg kg(-1) within 17 days after exposure of 100 mg kg(-1) melamine, and the carry-over rate for melamine from feed to the eggs was 1.21%. In the second experiment, melamine was detected in tissues within 3 days after exposure; the maximum concentration of melamine residues occurred in the 100 mg kg(-1) group and was as follows: egg (1.83) > kidney (1.21) > breast muscle (0.86) > liver (0.70) > serum (0.42). The melamine level in egg albumen was about twice that of egg yolk. Melamine levels in laying hens decreased rapidly with withdrawal from feed, but melamine only declined to undetectable levels in the egg at day 6 and in tissues at day 4 after last ingestion of 100 mg kg(-1). It can be concluded that a pathway exists for the transmission of melamine from feed to egg and body tissues and the carry-over rate of melamine is low, and that melamine is not metabolized into cyanuric acid in laying hens. A positive relationship exists between exposure levels and eggs or tissues, but no direct relationship between the exposure time and measured levels of melamine in eggs and tissues. The current Chinese limit for melamine in feed and feed material of 2.5 mg kg(-1) established on 8 June 2009 is sufficient to ensure that levels in eggs, breast muscle, liver as well as kidney will not exceed the limit of 2.5 mg kg(-1) in foods other than infant formula introduced by many countries including China.

Purification, characterization, and cloning of a novel phytase with low pH optimum and strong proteolysis resistance from Aspergillus ficuum NTG-23.

A novel phytase was isolated from Aspergillus ficuum NTG-23 with a procedure involving ion-exchange chromatography on DEAE-cellulose, CM-cellulose and FPLC-gel filtration on Superdex 75. The protein exhibited a molecular mass of 65.5kDa in gel filtration and SDS-PAGE. It possessed an optimal pH of 1.3 and an optimal temperature of 67 degrees C, and manifested a K(m) of 0.295mM and a V(max) of 55.9nmol (phosphate)/min. Phytase activity was not significantly affected by metal ions such as Ca(2+), Mg(2+), Mn(2+), Zn(2+), but was slightly stimulated in the presence of EDTA. The phytase was stable at 60 degrees C with no obvious loss of activity upon its incubation at 70 degrees C for 20min. The enzyme exhibited a broad substrate selectivity and showed strong resistance toward pepsin and trypsin. The unique properties suggest that the phytase has the potential to be useful as an animal feed supplement.

Effects of dietary taurine supplementation on growth performance and immune status in growing Japanese quail (Coturnix coturnix japonica).

The effect of dietary taurine was investigated on performance and immune responses in growing (1 to 42 d) Japanese quail. One-day-old quails of both sexes (300) were randomly allotted to 12 groups with 25 quails in each group (3 dietary treatments, 4 replicates). They were fed with maize-soybean meal-based diets containing 0, 0.01, or 0.05% taurine for 42 d. Dietary 0.01% taurine decreased food intake (P < 0.05) in the starter phase but did not affect BW gain and feed efficiency from d 1 to 42 of age. In the starter phase, dietary 0.05% taurine did not affect food intake but increased weight gain and improved feed efficiency (P < 0.05). In the grower phase, dietary 0.05% taurine did not affect weight gain but improved feed efficiency and decreased food intake (P < 0.05). In the overall period, dietary 0.05% taurine improved feed efficiency and decreased food intake (P < 0.05). The relative weights of thymus were greater in the quails given 0.01% taurine in diet (P < 0.05). The relative weights of the bursa of Fabricius and thymus were greater in dietary 0.05% taurine compared with the control (P < 0.05).The quail serum anti-Newcastle disease virus hemagglutination antibody titer in the taurine-supplemented diet had no significant effects compared with the control (P > 0.05). The response to phytohemagglutinin measured as the foot web index was significantly increased by 0.05% taurine supplementation (P < 0.05). It is concluded that the taurine-supplemented diet has a beneficial effect on immune responses and performance in growing Japanese quail.

Effect of polysavone (alfalfa extract) on abdominal fat deposition and immunity in broiler chickens.

Two hundred 1-day-old male commercial Arbor Acres broiler birds were randomly distributed to a control group and a polysavone group (5 replicates of 20 birds each) to investigate the influence of polysavone, a natural extract from alfalfa, on abdominal fat deposition and immunity in broiler chickens. Birds in the control group were supplied with a basal diet, and 0.06% polysavone was added to the basal diet of birds in the polysavone group. Body weight and feed consumption for each replicate were recorded weekly. At 3, 4, 5, and 6 wk of age, 4 birds from each replicate were randomly selected for blood and organ sampling. Polysavone had no significant effect on feed intake, BW, or feed:gain ratio in the experimental period, and it decreased the abdominal fat weights at 5 and 6 wk of age. Polysavone improved (P <0.05) the relative thymus and spleen weights at 6 wk of age and the bursa weights at 4 and 5 wk of age compared with the control group. At 4 and 6 wk of age, the proliferation of T and B lymphocytes in the polysavone group was significantly greater (P <0.05) than that in the control group. When birds were 4 and 5 wk of age, polysavone resulted in a significant increase (P <0.05) in serum anti-Newcastle disease virus hemagglutination inhibition antibody titer. These results showed that polysavone may decrease abdominal fat deposition and enhance immunity without an adverse effect on the performance of broiler chickens.

A new predictive and prognostic marker (ATP bioluminescence and positron emission tomography) in vivo and in vitro for delivering adjuvant treatment plan to invasive breast tumor patients.

The cell proliferation rate has been used to assess the biological aggressiveness and the metastatic potential of breast carcinoma. Different methods (flow cytometric S phase and proliferation associated antigens) have been used to assess the rate of proliferation previously. In this preliminary study, the cell proliferation rate of normal (N=45), benign (N=29) and invasive breast tumor tissue (N=70) has been quantified in vitro by ATP bioluminescence assay. Next, individual prognostic factor (tumor grades, lymph node involvement, estrogen and progesterone receptor and HER-2 status) has been correlated with the level of metabolic rate (ATP). The results showed that invasive tumor had the highest level of ATP bioluminescence compared with that of benign tumor (mean difference=1.97) and the normal breast tissue (mean difference=2.75). In addition, ATP level positively correlated with the number of axillary lymph node involvement (r(spearman)=0.433, P=0.021). These findings suggested that the measurement of ATP level may serve as a mean for the detection of cell proliferation and hence a surrogate marker for disease prognosis.

Prevalence of workplace violence against nurses in Hong Kong.

OBJECTIVES: To determine the prevalence and nature of workplace violence against nurses, and how nurses deal with such aggression; and to identify the risk factors related to violence in the hospital environment. DESIGN: Cross-sectional questionnaire study. SETTING: University teaching hospital, Hong Kong. PARTICIPANTS: All nursing staff in the hospital, except nurses who were unable to read Chinese or who did not have patient contact (eg those worked in administrative positions), were invited to complete a questionnaire. MAIN OUTCOME MEASURES: Demographic data of the respondents, incidence of and risk factors contributing to workplace violence. RESULTS: A total of 420 nurses returned the completed questionnaire (response rate, 25%). Three hundred and twenty (76%; 95% confidence interval, 72-80%) nurses reported abuse of any kind--verbal abuse, 73%; bullying, 45%; physical abuse, 18%; and sexual harassment, 12%. Most (82%) nurses who experienced verbal abuse tended to confide in friends, family members, or colleagues. Some (42%) ignored the incident. Risk factors for workplace violence included: working in male wards and in certain specialties such as the Accident and Emergency Department, Community Nursing Service, and the Orthopaedics and Traumatology Department. CONCLUSION: Workplace violence against nurses is a significant problem in Hong Kong. Further large-scale studies should be conducted to more closely examine the problem.