Search

The study was designed to investigate productivity parameters, egg quality and the cost effectiveness of feeding laying hens with diets where ground wheat was replaced with varying levels of whole hybrid rye grain cv. Brasetto. A total of 396 Hy-Line Brown hens at 34 weeks of age were allocated to three treatment groups with 66 replicates (cages) of two hens each, and were fed isocaloric and isonitrogenous pelleted diets for 12 weeks. The diet for control group hens (R0) did not contain rye grain. In diets for hens from groups R100 and R200, ground wheat grain was partially replaced with whole rye grain at 100 g/kg and 200 g/kg, respectively. Whole intact rye grains were included in the diets. The cost of diets containing 10% and 20% of whole rye grain was by approximately 1.0% and 1.9% lower, respectively, compared with the control diet that contained no rye grain. During the entire 12-week feeding period, dietary treatments had no effect on laying rate (95.64% to 97.07%), egg weight (62.51 g to 62.74 g) or egg mass output (5.04 kg to 5.12 kg per hen). The inclusion of whole rye grain in layer diets caused a linear increase in feed intake (P

This study was conducted to determine the effect of probiotic Pediococcus acidilactici (PA) strain MA18/5M supplementation of diets with different dietary energy levels on productive performance, egg quality, and body composition in Hy-Line Brown hens during a 16-week period from 32 to 47 wk of age. The experimental treatments with a 2 × 2 factorial design received a 2 wheat–corn–soybean diet: a moderately low energy density diet with 2,650 kcal ME/kg (M-LED) and a low energy density diet based on the M-LED diet with 2,550 kcal ME/kg (LED), each diet without and with probiotic supplementation (M-LED, LED, M-LEDp, and LEDp, respectively). Reduced dietary energy levels had a particularly negative effect on egg weight (61.7 vs. 63.3 g; −2.6%, P

The research hypothesis postulated that the optimal dietary inclusion levels and ratios of lysine (Lys), arginine (Arg), and methionine (Met) can increase the growth potential of hybrid turkeys and limit metabolic disorders that weaken immune function. The experiment was carried out in a full rearing cycle, from 1 to 16 wk of age, in a two-factorial randomized design with 3 levels of Arg and 2 levels of Met (90, 100 and 110% of Arg, and 30 or 45% of Met, relative to the content of dietary Lys), with 6 groups of 8 replicates per group and 18 turkeys per replicate. In the first and second month of rearing, a significant dietary Arg-by-Met interaction was noted for daily feed intake and body weight gain, and a more beneficial effect was exerted by higher Met content and medium Arg content. Throughout the experiment, the higher dietary Met level increased the final body weight (BW) of turkeys (P = 0.001). Different dietary Arg levels had no influence on the growth performance of turkeys, but the lowest level decreased dressing yield (P = 0.001), and the highest level increased the percentage of breast muscles in the final BW of turkeys (P = 0.003). The lowest Arg level (90% of Lys content) undesirably increased the concentration of the proinflammatory cytokine IL-6 (P = 0.028) and decreased globulin concentration (P = 0.001) in the blood plasma of turkeys. The higher dietary Met level (45% of Lys content) increased plasma albumin concentration (P = 0.016). It can be concluded that higher dietary levels of Met (45 vs. 30% of Lys content) and Arg (100 and 110 vs. 90% of Lys content) have a more beneficial effect on the growth performance and immune status of turkeys.

We surveyed European medical schools regarding teaching of prudent antibiotic prescribing in the undergraduate curriculum. We performed a cross-sectional survey in 13 European countries (Belgium, Croatia, Denmark, France, Germany, Italy, Netherlands, Norway, Serbia, Slovenia, Spain, Switzerland, United Kingdom) in 2013. Proportional sampling was used, resulting in the selection of two to four medical schools per country. A standardized questionnaire based on literature review and validated by a panel of experts was sent to lecturers in infectious diseases, medical microbiology and clinical pharmacology. In-depth interviews were conducted with four lecturers. Thirty-five of 37 medical schools were included in the study. Prudent antibiotic use principles were taught in all but one medical school, but only four of 13 countries had a national programme. Interactive teaching formats were used less frequently than passive formats. The teaching was mandatory for 53% of the courses and started before clinical training in 71%. We observed wide variations in exposure of students to important principles of prudent antibiotic use among countries and within the same country. Some major principles were poorly covered (e.g. reassessment and duration of antibiotic therapy, communication skills). Whereas 77% of the respondents fully agreed that the teaching of these principles should be prioritized, lack of time, mainly due to rigid curriculum policies, was the main reported barrier to implementation. Given the study design, these are probably optimistic results. Teaching of prudent antibiotic prescribing principles should be improved. National and European programmes for development of specific learning outcomes or competencies are urgently needed. Clinical Microbiology and Infection © 2014 European Society of Clinical Microbiology and Infectious Diseases. Published by Elsevier Ltd. All rights reserved

The objective of this study was to determine the physiological effect of supplementing turkey diets with different vegetable oils, i.e. soyabean (S) oil, rapeseed (R) oil and linseed (L) oil, which have different concentrations of polyunsaturated fatty acids (PUFA). The total contents of n-6 PUFA and n-3 PUFA were as follows: diet S, 49.7 and 7.02%, diet R, 33.0 and 7.96%, and diet L, 30.3 and 30.6%; the n-6/n-3 PUFA ratio was 7.3, 4.4 and 1.0, respectively. The experimental materials comprised four groups of turkey hens (14 replications of fifteen birds each). The amount of dietary oil supplementation was increased from 2% in the first four weeks to 6% at the final stage of fattening (14-15 weeks). The apparent total tract digestibility of crude fat in 7-week-old turkeys (2.5% oil in diet) ranged from 89.9 to 91.1%, and showed no statistically significant differences. Dressing percentage, breast, thigh, and drumstick muscle contents did not differ among groups. Group L had a significantly higher intestinal fat weight (0.84% body weight (BW)) than the other groups (0.60-0.62%). In comparison with diet S, diets R and L contributed to a significant decrease in the levels of saturated fatty acids (from 32.9% to 30.3 and 30.2%) in breast meat lipids. In group L turkeys, the share of PUFA in total fatty acids in breast meat lipids was significantly higher (37.3%

The objective of this study was to determine the effect of genotype (slower-growing vs. fast-growing) and production system (access to outdoors vs. indoor) on the growth performance, carcass yield and meat quality (chemical composition, physicochemical and sensory properties) of chickens. The experiment was performed on 1,040 day-old hybrid male chickens of two genotypes. Slower-growing chickens (Hubbard JA957, certified) and fast-growing chickens (Hubbard F15) were fed identical diets until 65 days of age. Both genotypes (each represented by 520 birds) were divided into two subgroups and were raised in pens on litter with outdoor access or in indoor confinement without outdoor access (four replications per subgroup, each of 65 birds). Until day 21, the birds stayed in the indoor facility, in deep-litter pens. The birds could forage on pasture 12 h daily, commencing at three weeks of age. Stocking density was 0.13 m 2 floor space per bird in pens on litter, and 0.8 m 2 per bird in grassy yards. Compared with fast-growing, slower-growing chickens were significantly lighter (by 17%), had a lower breast and thigh muscle yield and a higher abdominal fat content, but they were characterized by higher survival rates at 65 days, a higher protein content and a lower fat content of breast meat. Outdoor access had no negative effects on the growth performance, muscle yield, the fatty acid profile and oxidative status of meat lipids. The meat of free-range chickens was darker in color, it had a higher protein content and a better water-holding capacity, but it was less juicy than the meat of birds raised indoors. (Key Words : Slower-growing Chickens, Outdoor Access, Growth Performance, Carcass Traits, Meat Quality)

(Received 6 October 2008; revised version 21 April 2009; accepted 6 November 2009)ABSTRACTThe aim of this study was to examine the effect supplementation of diet with 20 mg Cu/kg from copper sulphate (Cu-SUL) and/or copper amino acid chelate (Cu-AA) on the copper balance, bone mineralization status and growth performance of turkeys. A total of 840 one-day-old heavy Large White BIG-6 turkey males were randomly assigned to 4 groups: Cu-0, Cu-SUL, Cu-SUL/Cu-AA and Cu-AA. The results of the present study confirm the Cu-requirements for growing/fattening turkeys given by scientific bodies (e.g., NRC, INRA, GfE). The Cu unsupplemented basal diet contained 11.3 mg Cu/kg is enough for turkeys and further Cu-supplementations are not necessary. Further supplementations from inorganic and/or organic sources did not significantly improve turkeys performance and health. Plasma superoxide dismutase activity, physico-chemical properties of bone and the concentrations of Ca and P in the tibial bone were generally not affected by dietary treatment. Further Cu supplementation from inorganic and/or organic sources increase the Cu-excretion (from 808 to 1915-2164 mg per animal) and may cause additional environmental problems. KEY WORDS: copper, chelate, bone mineralization, performance, turkeys

The aim of this study was to evaluate the effects of a different dietary content of sweet yellow lupine seed meal (YLM) on gastrointestinal tract development and function, the growth performance (13-18 weeks of age) and meat quality of growing-finishing turkeys. Control grower and finisher diets contained soybean meal (SBM), and in experimental diets, SBM was replaced with YLM at 6%, 12% and 18% (Groups L6, L12 and L18, respectively). The diets were formulated to be iso-nitrogenous and iso-energetic. In the first phase of feeding, YLM tended to decrease feed intake (p = 0.087) and body weight gain (BWG, p = 0.078) linearly due to significant deterioration in feed conversion ratio (FCR, p = 0.030). An opposite trend was noted in the second phase of feeding: BWG increased (p = 0.069) and FCR improved significantly (p = 0.004). Over the entire experiment, inclusion of YLM did not affect feed intake, BWG, FCR or excreta dry matter (DM) content. The highest YLM dietary level had no adverse effects on fermentation processes in the caeca. Positive changes, which were observed in turkeys fed YLM-supplemented diets, included an increased gizzard weight, a decreased pH of gizzard contents and a decreased viscosity of small intestinal digesta. The inclusion of YLM at 18% had no effect on carcass quality, the pH and colour intensity of breast meat. The highest inclusion rate of YLM (18%) in turkey diets significantly (p0.05) intensified the fatty flavour of meat and increased its hardness, springiness and chewiness, whereas it had no effect on the appearance, aroma and overall acceptability of breast meat. It can be concluded that YLM at the highest dietary rate used in the present experiment (18% of the diet) had no adverse effects on the growth performance or the analysed physiological parameters of turkeys.

The objective of this study was to determine the effect of dietary probiotic Pediococcus acidilactici (PA) strain MA18/5M on performance, egg traits, egg cholesterol content, and fatty acid composition in laying hens during a 24-wk period. A total of 222 Hy-Line Brown laying hens, 22 wk of age, were divided into 3 treatment groups. Control group (C) hens were fed a basal diet with no probiotic added. In group PA1, the basal diet was supplemented with PA at 100 mg.kg(-1) of feed for the first 12 wk and 50 mg.kg(-1) feed for the next 12 wk, whereas treatment PA2 was supplemented with 100 mg.kg(-1) feed for the whole trial period. Dietary treatments did not significantly affect the BW, feed intake, and egg production of hens. Pediococcus acidilactici supplementation increased egg weight (P < 0.05), eggshell thickness, eggshell relative weight, and egg specific gravity, and it improved feed efficiency ratio per kilogram of eggs (P < 0.01). Moreover, PA dietary supplementation resulted in a significant (P < 0.05) decrease in the number of broken eggs and eggs without the shell, leading to a significant (P < 0.01) reduction in the number of downgraded eggs (39% for PA1 and 52% for PA2). After 6 mo of probiotic supplementation, significant differences were also found in the fatty acid composition and cholesterol content of egg yolk. The yolk cholesterol content, regardless of PA dose, decreased by more than 10%. The concentrations of total polyunsaturated fatty acids, including linoleic acid and linolenic acid, were significantly higher in treatment PA2 (6.5% increase) than in C and PA1. In conclusion, dietary supplementation of Pediococcus acidilactici MA 18/5M at 100 mg.kg(-1) has potential commercial applications for improvements in hen performance and eggshell quality during the early laying period.

Bacteremia is a well known cause of morbidity and mortality in hematopoietic stem cell transplant (HSCT) recipients and enterococci are among the most frequently isolated pathogens. The aim of this study was to identify risk factors for enterococcal bacteremia during the first 30 days after allogeneic HSCT. A retrospective case-control study was performed; for each case, 3 controls were randomly selected among 306 patients transplanted during the study period (January 1, 2004 to December 31, 2007). Odds ratios (OR) with 95% confidence intervals (CI) were calculated for variables influencing the risk for bacteremia. Overall, 33 patients developed enterococcal bacteremia, within a median of 9 days after HSCT (range, 2-24). The cumulative incidence was 10.8%. Multivariate analysis identified the following variables as risk factors for enterococcal bacteremia: donor and transplant type (greater risk for mismatched related or cord blood) (OR=8.98, 95% CI, 1.65-48.99 and OR=7.52, 95% CI, 1.56-36.31, respectively, P=0.047); severe (grades 3-4) mucositis (OR=9.04, 95% CI, 1.97-41.52, P=0.018); pharyngeal enterococcal colonization (OR=4.48, 95% CI, 1.11-18.03, P=0.035); and previous empirical therapy with cephalosporins (OR=4.16, 95% CI, 0.93-18.66 for 1-7 days of therapy, and OR=7.31, 95% CI, 1.78-30.12 for 8-23 days, P=0.018). Higher Karnofsky score (≥50) and previous empirical therapy with glycopeptides were associated with a decreased risk (OR=0.25, 95% CI, 0.06-0.97, P=0.045 and OR=0.11, 95% CI, 0.02-0.59, P=0.010, respectively). The crude mortality at 7 and 30 days was 12% (4/33) and 24% (8/33), respectively. Enterococcal bacteremia is frequent after allogeneic HSCT. The factors associated with this infection are type of transplant, pharyngeal colonization, severe mucositis, and use of cephalosporins. Good general conditions and the use of vancomycin were associated with lower risk of enterococcal bacteremia.

Since 2002, blood donors in Poland have been tested not only for hepatitis C virus antibodies (anti-HCV) but also for HCV RNA or HCV core antigen. This screening program identifies asymptomatic, recently infected individuals with no anti-HCV (in the "window period"). The aim of this study was to compare HCV genotype and subtype distribution in window-period (wp) donors, anti-HCV-positive donors, and chronic hepatitis C (CHC) patients.A total of 2.37 million donors were investigated for HCV RNA, and 340,000 for HCV core antigen. HCV genotypes and subtypes were investigated in 50 HCV RNA-positive, anti-HCV-negative donors; in 70 anti-HCV-positive donors; and in 170 CHC patients. Re-questioning of wp donors for probable risk factors was introduced.HCV RNA was detected in 50 donors of 2.71 million (1:54,200) anti-HCV-negative blood donations. Of these 50 donors, 36 percent exhibited Subtype 1b, whereas Subtypes 3a and 4c/d were identified in 40 and 14 percent, respectively. In anti-HCV-positive donors and CHC patients, the frequency of Subtype 1b was significantly higher (75.7 and 85.3%, respectively); in both groups the lower frequency of Subtypes 3a (14.3 and 10.6%, respectively) and 4c/d (4.3 and 1.2%, respectively) was found. The probable source of infection was identified in 9 wp donors.The frequency of wp donors is 18.5 per 1 million. The unexpected high frequency of Genotype 4 and Subtype 3a and the low frequency of Subtype 1b was observed in wp donors compared to anti-HCV-positive individuals. Additional epidemiologic questioning introduced after HCV RNA detection may help to identify infection source.

Neonates from multiple pregnancies were evaluated taking into account their condition in the 1st and 5th minutes after birth, their birth weight, time which they spent in the neonatal department and occurrence of complications during the adaptation period, such as respiratory insufficiency, intrauterine infection and hyperbilirubinaemia. On the basis of our research, we think that multiple pregnancy requires particular diagnostics and individualisation of prenatal care. We believe also that neonates from such pregnancies are particularly exposed to development of a severe form of diverse complications. In our investigations, we observed a beneficial influence of breast-feeding--from the first hours of life--on the course of adaptation of neonates. However, we think that confirmation of these findings requires more detailed investigations.

Multiple pregnancies pose numerous threats both for foetuses and neonates. The following disorders are encountered among the most important perinatal disturbances: EPH – gestosis, immature labour and premature rupture of foetal membranes. Also during the labour, particularly the second one and the following ones, various lesions that may cause increased incidence of diseases and mortality in this group of neonates [5, 6, 10, 18, 21, 22, 25]. Complications encountered in neonates from multiple pregnancies, resulting both from abnormal conditions of intrauterine development and from perinatal pathology, require still wider studies, particularly within the context of intensified obstetrical care of women with multiple pregnancies [7, 11, 13, 16, 24].In the years 1996-1998, out of 3883 neonates born in the Chair of Obstetrics and Gynaecology of the Silesian Medical Academy, 152 (3,9%) came from multiple pregnancies. This period included one case of quadruplets, eight cases of triplets and 62 twin pregnancies. Pregnant women were directed to our Chair within the program of multidisciplinary care of women with complicated pregnancies and they also came from the Department of Pathology of Pregnancy in our Chair. In 18 women, treated for infertility, the pregnancy was accomplished as a result of application of various techniques of assisted procreation. In one case – 4 foetuses were indicated, in five cases – three foetuses and in 12 cases – two foetuses.

Diabetic fetopathy is a complex the pathophysiology of which is not all that clear and that is why all neonates whose mothers develop diabetes demand, regardless of their body weight, from the first hours of their live an intensive neonatal care. In the period from 1993 to 1998 in the Chair of Obstetrics and Gynaecology in Bytom 237 babies were born in whom, in the first day of their live, clinical and biochemical screening was carried out in order to determine degree of diabetic fetopathy progression. Summary results of six years care of newborns, born by diabetic mothers. The following parameters were measured: morphometric parameters, course and complications of adaptation processes.

The authors analysed a way of pregnancy ending and indications to cesarean section in group 256 diabetic pregnant women. We estimated that percentage for cesarean sections was 51.56%. We noticed downward tendency of elective indications to cesarean section (the most common was gestosis) and upward tendency of intrapartum indications (the most common was fetal distress syndrome). The percentage of cesarean sections in diabetic pregnant women group in our study is similar to results from the other centres.

Authors had retrospectively analyzed basic exponents of perinatal care of 256 pregnant diabetic woman in years 1993-1998 hospitalized and delivered in Clinic of Obstetrics and Gynaecology in the town of Bytom. Our results were very similar to the other of Referent Centres Of Technical Methods Bank Used in Diagnostic and Therapy of Diabetes: cesarean sections percentage 51.56%, 4.7% of undelivered pregnancies, 2.34% of new-born delivered with multiple congenital defaults, 17.2% of macrosomies and 39.8% pre-term deliveries were noted. Disadvantage perinatal effects in diabetic women might be reduced by very early diagnosis of diabetic and intensive biophysical and biochemical fetal care, which is strongly suggests by the authors.

Diabetic fetopathy is a complex the pathophysiology of which is not all that clear and that is why all neonates whose mothers develop diabetes demand--regardless of their body weight--from the first hours of their live an intensive neonatal care. In the period from 1993 to 1997 in the Chair I of the Hospital of Obstetrics and Gynaecology in Bytom 191 babies were born in whom--in the first day of their live clinical and biochemical screening was carried out in order to determine degree of diabetic fetopathy progression. Summary results of five years care of newborn, born by diabetic mothers. The following parameter were measured: morphometric parameters, the course of adaptation processes, congenital defect occurrence, morbidity and motility in this group of newborn.

Authors had retrospectively analysed basic exponents of perinatal care of 199 pregnant diabetic woman in years 1993-1997 hospitalised and delivered in The First Clinic of Obstetrics and Gynaecology in the town of Bytom. Our results were very similar to the other Referent Centres Of Technical Methods Bank Used in Diagnostic and Therapy of Diabetes: average gestation age 38 +/- 1 hbd, caesars sections percentage 53.4%, 5.5% of undelivered pregnancies, 0.5% of new-born delivered with multiple congenital defaults, 16.1% of macrosomies and 28.1% pre-term deliveries were noted. Disadvantage perinatal effects in diabetic women might be reduced very early diagnosis of diabetic and intensive biophysical and biochemical fatal care, which is strongly suggests by the authors.

Summary The effects of oxygen concentration on 14 CO 2 uptake, the time required to compensation of respiration by photosynthesis and the greening of etiolated seedlings of bean upon illumination were investigated. It has been found that this factor had a large inhibitory action on the processes examined. In atmospheres of 1 and 21 % O 2 the fixation of 14 CO 2 was observed after 3 hours of illumination, while in 100% O 2 after 8 hours. The time required for compensation of respiration by photosynthesis was in order of 8 hours in 1% O 2 , 10 hours in 21 % O 2 and 20 hours in 100 % O 2 respectively. The chlorophyll a + b biosynthesis followed the similar pattern of inhibition by O 2 as photosynthesis. The lag phase for the appearance of chlorophyll biosynthesis in 1 and 21 % O 2 was 3 hours while in 100%O 2 6 hours.

Summary The effects of either 0.5 or 10 hours of pretreatment of bean leaves with 100 % O2 and illumination on 14CO2 uptake and on photosynthetic 14C-products after 2 minutes of fixation were examined. For comparative purposes these measurements were carried out after illumination of the leaves for 0.5 hours in 21 % O2. It was found that the 14CO2 uptake of leaves having been exposed to 100 % O2 for 0.5 or 10 hours was inhibited by 76.2 and 96.5 % respectively compared to that after pretreatment in air. The chlorophyll a and b contents in the leaves of plants pretreated with 100 % O2 for 10 hours was about 2.5 times lower than that of plants pretreated with air or with 100 % O2 for 0.5 hours. High O2 concentration treatments of plants caused a decrease in the incorporation of 14C into the labelling of the amino acid fraction. In the amino acids the bulk of the radioactivity was found in glycine and serine. The incorporation of radioactivity into these compounds was enhanced by pretreatment with 100 % O2. At the same time, the labelling of alanine and of glutamic and aspartic acids was lower compared to that after pretreatment in air. In the organic acid fraction the greatest part of the radioactivity was found in malate and was found in greater amounts after exposure to an atmosphere of 100 % O2. Glycolic acid was found after exposure to air and was absent after pretreatment 100 % O2. Among the sugars, sucrose was a major radioactive product either in air or in 100 % O2. A high O2 concentration and a treatment of the leaves in this atmosphere for 10 hours enhanced the incorporation of 14C into this compound. It is suggested that high O2 concentrations enhanced the glycolic acid pathway and inhibited the biosynthesis the amino acids derived from PGA. A high O2 may also stimulate the production of sucrose in this manner.

The objective of this study was to determine the effect of moderate dilution (gradually up to 22.5%) of standard diets (C) with wheat in different physical forms, ground and pelleted (GW) or whole (WW), on growth rate, feed conversion ratio (FCR), carcass yield, and incidence of footpad dermatitis (FPD) in growing heavy-type turkeys. Dietary treatments in which part of the basal diet (control, C) was replaced with a low, medium, or high wheat content (8.2%, 13.3%, or 18.2% on average in groups W L , W M , and W H , respectively) were compared. Seven experimental groups consisted of six replicates each (n=30 birds) fed from 4 to 18 weeks of age. After four weeks of experimental feeding, groups GW L and WW L , relative to group C, were characterized by higher coefficients of dry matter digestibility (P=0.003), apparent nitrogen retention (P=0.096), and energy metabolizability (P=0.003), as well as by a higher metabolizable energy content of the daily ration (P

Purpose We compared the risk factors, the diagnostic tools and the outcome of filamentous fungal infections (FFIs) in hematological patients (HAEs) and non-hematological patients (non-HAEs). Methods Prospective surveillance (2009–2011) of proven and probable FFIs was implemented in 23 Italian hospitals. Results Out of 232 FFIs, 113 occurred in HAEs and 119 in non-HAEs. The most frequent infection was invasive aspergillosis (76.1 % for HAEs, 56.3 % for non-HAEs), and the localization was principally pulmonary (83.2 % for HAEs, 74.8 % for non-HAEs). Neutropenia was a risk factor for 89.4 % HAEs; the main underlying condition was corticosteroid treatment (52.9 %) for non-HAEs. The distribution of proven and probable FFIs was different in the two groups: proven FFIs occurred more frequently in non-HAEs, whereas probable FFIs were correlated with the HAEs. The sensitivity of the galactomannan assay was higher for HAEs than for non-HAEs (95.3 vs. 48.1 %). The overall mortality rate was 44.2 % among the HAEs and 35.3 % among the non-HAEs. The etiology influenced the patient outcomes: mucormycosis was associated with a high mortality rate (57.1 % for HAEs, 77.8 % for non-HAEs). Conclusions The epidemiological and clinical data for FFIs were not identical in the HAEs and non-HAEs. The differences should be considered to improve the management of FFIs according to the patients’ setting.