Effect of Quantum phytase on nutrient digestibility and bone ash in White Leghorn laying hens fed corn-soybean meal-based diets.

The efficacy of an Escherichia coli 6-phytase supplementation (Quantum) on nutrient digestibility-retention and bone ash in laying hens fed corn-soybean meal (CSM) diets was investigated. White Leghorn hens (Shaver and Bovan strains) were fed CSM diets containing 0.35% (positive control, PC), 0.25% (negative control 1, NC1), or 0.15% (negative control 2, NC2) nonphytate P from 21 to 61 wk of age. Six more diets were manufactured by supplementing the negative control diets with 200, 400, and 600 units per kilogram of exogenous phytase resulting in a total of 9 treatments. Each dietary treatment x strain subclass was replicated twice with 6 hens per replication. Fecal and ileal digesta samples were collected at 42 wk of age to determine apparent nutrient digestibility or retention. Left tibiae were collected at 42 and 61 wk of age to determine bone ash. The coefficients for ileal digestibility and fecal retention for protein were higher (P < 0.05) for the unsupplemented negative control treatments compared with the PC. A linear reduction in phytate digestibility and ileal protein digestibility was reported with increasing levels of phytase to the NC1 diet. Phytase addition to the NC1 treatment resulted in a linear decrease in the digestibility of amino acids except for methionine and proline. Significantly higher phytate digestibility was demonstrated with the NC2 treatment containing 400 units per kilogram of phytase compared with the PC. Tibial bone ash percentage was higher (P < 0.05) in 61-wk-old hens fed 200 or 400 units per kilogram of phytase-supplemented NC2 diets. Significantly higher diet AME and fecal protein retention were demonstrated for Shaver hens in comparison to the Bovan hens. Overall, the Quantum phytase was not efficacious at improving nutrient digestibility-retention in laying hens fed CSM diets deficient in nonphytate P.

The efficacy of quantum phytase in a forty-week production trial using white leghorn laying hens fed corn-soybean meal-based diets.

Microbial phytase is a prominent feed enzyme used in animal feeds, but there is relatively little information on its use in laying hen diets. In this experiment, an Escherichia coli 6-phytase (Quantum) was evaluated for its efficacy in a 40-wk laying hen production trial. A total of 1,080 White Leghorn hens (540 each of Shaver and Bovan strains) were fed mash corn-soybean meal-based diets containing 0.35% (positive control, PC), 0.25% (negative control, NC1), or 0.15% (NC2) nonphytate phosphorus (NPP). Six more diets were manufactured by supplementing the negative control diets with 200, 400, and 600 U/kg of exogenous phytase, resulting in a total of 9 treatments. Each dietary treatment x strain subclass was replicated 4 times with 5 adjoining cages per replicate (3 hens per cage) in a randomized complete block design. Production performance was measured from 21 to 61 wk of age. Only minor differences in production characteristics were found between the PC and NC1 treatments regardless of phytase addition, indicating that 0.25% NPP resulted in P intake that was at or above the hen's requirement. In contrast, the hens fed 0.15% NPP diet without phytase supplementation had significantly (P < 0.05) reduced total hen housed egg production and body weight at 61 wk of age in comparison to the PC treatment, whereas the incidence of soft-shelled, cracked, and broken eggs was increased significantly (P < 0.05) in hens fed the NC2 diet. Addition of phytase to the NC2 diet improved these production characteristics to levels equal or better than the PC diet. The results indicated that Quantum phytase was efficacious in corn-soybean meal-based diets fed to White Leghorn laying hens and can be used to reduce diet supplementation with inorganic phosphorus.

Spherical navigator registration using harmonic analysis for prospective motion correction.

Spherical navigators are an attractive approach to motion compensation in Magnetic Resonance Imaging. Because they can be acquired quickly, spherical navigators have the potential to measure and correct for rigid motion during image acquisition (prospectively as opposed to retrospectively). A limiting factor to prospective use of navigators is the time required to estimate the motion parameters. This estimation problem can be separated into a rotational and translational component. Recovery of the rotational motion can be cast as a registration of functions defined on a sphere. Previous methods for solving this registration problem are based on optimization strategies that are iterative and require k-space interpolation. Such approaches have undesirable convergence behavior for prospective use since the estimation complexity depends on both the number of samples and the amount of rotation. We propose and demonstrate an efficient algorithm for recovery of rotational motion using spherical navigators. We decompose the navigator magnitude using the spherical harmonic transform. In this framework, rigid rotations can be recovered from an over-constrained system of equations, leading to a computationally efficient algorithm for prospective motion compensation. The resulting algorithm is compared to existing approaches in simulated and actual navigator data. These results show that the spherical harmonic based estimation algorithm is significantly faster than existing methods and so is suited for prospective motion correction.

Methionine and cystine requirements of slow- and fast-feathering broiler males from three to six weeks of age.

Two experiments were conducted to first determine Met then Cys needs of broilers from 3 to 6 wk of age and whether differences existed between slow-feathering (Ross x 308) and fast-feathering (Ross x 3F8) males. A corn-soybean meal diet (20.0% CP; 3,150 kcal ME/kg) with graded levels of Met or Cys was offered. The first experiment had dietary Met levels of 0.32, 0.38, 0.44, and 0.50% with surfeit Cys (0.40%). Broilers from both feathering strains responded similarly to supplemental Met. Although body weight was not responsive, F/G improved through to the highest level of dietary Met (linear, P < 0.05). Chilled carcass weight increased with Met (linear, P < 0.05) paralleling F/G; however, no differences were detected in the amount of associated abdominal fat. Breast fillet yield increased with Met to maximize at 0.48% (quadratic, P < or = 0.009). In a satellite study using the same birds in cages and feeds, N retention at d 29 maximized at 0.46% Met (quadratic, P < 0.05). The second experiment had Cys at 0.32, 0.34, 0.38, and 0.46% with Met fixed at a submarginal level of 0.38%. Increasing dietary Cys had no effect on live performance of slow-feathering birds, whereas weight gain of fast-feathering birds achieved maximum at 0.36% Cys (cubic; P < 0.05) with F/G responding similarly. Chilled carcass (cubic, P < 0.002) and breast fillet weights (cubic, P < 0.001) of fast-feathering birds also increased with Cys to maximize at 0.36%, and the amount of abdominal fat was not influenced by feathering or Cys supplementation. Separate measurement of N retention at d 31 failed to detect a difference in protein utilization attributable to feathering, but an optimum was achieved at 0.40% Cys with both broiler sources. Overall results suggest that the Met requirement for broiler males between 3 and 6 wk of age was independent of feathering and approximated 0.46% (95% of the level of maximal response). Cystine requirements once corrected for submarginal Met status indicated a greater demand by fast- than slow-feathering male broilers corresponding to 0.42 and 0.37%, respectively.

Automatic segmentation of the colon for virtual colonoscopy.

Virtual colonoscopy is a minimally invasive technique that enables early detection of colorectal polyps and cancer. Normally, a patient's bowel is prepared with colonic lavage and gas insufflation prior to computed tomography scanning. An important step for 3D analysis of the image volume is segmentation of the colon. The high-contrast gas/tissue interface that exists in the colon lumen makes segmentation of the majority of the colon relatively easy; however, two factors inhibit automatic segmentation of the entire colon. First, the colon is not the only gas-filled organ in the data volume: lungs, small bowel, and stomach also meet this criterion. User-defined seed points placed in the colon lumen have previously been required to spatially isolate the colon. Second, portions of the colon lumen may be obstructed by peristalsis, large masses, and/or residual feces. These complicating factors require increased user interaction during the segmentation process to isolate additional colonic segments. To automate the segmentation of the colon, we have developed a method to locate seed points and segment the gas-filled lumen sections without user supervision. We have also developed an automated approach to improve lumen segmentation by digitally removing residual contrast-enhanced fluid. Experimental results with 20 patient volumes show that our method is accurate and reliable.

Enzyme supplementation and particle size of wheat in diets for nursery and finishing pigs.

Three experiments were conducted to determine the effects of enzyme supplementation and particle size of wheat-based diets on growth performance and nutrient digestibility in nursery and finishing pigs. In Exp. 1, 180 weaned pigs (5.7 kg and 21 d of age) were fed diets in a 35-d growth assay without or with a Trichoderma longibrachiatium enzyme product (4,000 units of xylanase activity per gram of product) and with wheat ground to mean particle sizes of 1,300, 600, or 400 microm. Enzyme supplementation had no effect on ADG or gain/feed (P > 0.32), but there was a trend (P < 0.10) for greater digestibility of DM (d 6) in enzyme-supplemented diets. A particle size of 600 microm supported the greatest overall gain/feed (quadratic effect, P < 0.01). An interaction of enzyme supplementation with particle size occurred; gain/feed was improved (P < 0.01) with enzyme supplementation at the coarse (1,300 microm) particle size but not when the wheat was ground to 600 or 400 microm. In Exp. 2, 160 finishing pigs (67 kg) were fed a diet without or with the same enzyme used in Exp. 1 and wheat ground to 1,300 or 600 microm. No interactions occurred between enzyme supplementation and particle size of the wheat (P > 0.15). However, there were trends for greater gain/feed (P < 0.10) during the 67- to 93-kg phase of the experiment and for greater digestibility of DM (P < 0.10) and N (P < 0.07) with enzyme supplementation. When particle size was reduced from 1,300 to 600 microm, gain/feed was improved (P < 0.03) for the 93- to 114-kg phase of the growth assay, and digestibilities of DM (P < 0.02) and N (P < 0.04) were greater. In Exp. 3, 160 finishing pigs (63 kg) were given diets without or with the enzyme product and wheat ground to 600 or 400 microm. Enzyme supplementation improved ADG (P < 0.04) in the 90- to 115-kg phase but otherwise did not affect growth performance, carcass measurements, or stomach lesions. A particle size of 400 microm increased overall gain/feed (P < 0.04), digestibilities of DM and N (P < 0.01), and development of stomach lesions (P < 0.01). In conclusion, pigs did not benefit consistently from enzyme supplementation. However, wheat particle sizes of 600 and 400 microm supported the best overall performance in nursery and finishing pigs, respectively.

Organochlorine residues in fish oil dietary supplements: comparison with industrial grade oils.

The market for fish oils as dietary supplements is of global importance. Although it is widely recognised that lipophilic organic chemicals, particularly organochlorines, can accumulate in fish oils, dietary supplements are not routinely considered when estimating average daily intakes for these contaminants. This paper reports levels of organochlorine residues in 44 fish oils, collected from 15 countries between 1994 and 1995, including 38 purchased over the counter as dietary supplements. Despite controls on the use of persistent organochlorine substances, appreciable quantities are found in oils sold as dietary supplements. Levels are discussed in relation to the significance of fish oil dietary supplements as contributors to daily intake of PCBs and pesticide residues.

Diorthosubstituted polychlorinated biphenyls in caudate nucleus in Parkinson's disease.

As it had previously been demonstrated that there were reduced brain dopamine concentrations in monkeys who had been given polychlorinated biphenyls (PCBs) chronically, we hypothesized that organochlorine compounds in general, and PCBs in particular, might be important in the pathogenesis of Parkinson's disease (PD). In a study of caudate nucleus obtained post mortem from patients with Parkinson's disease and from controls, there were significantly higher concentrations of the organochlorine insecticide dieldrin and the PCB congener 153 in the PD tissue. DDE, PCB congener 180, and total PCBs (matched with a commercial preparation) also tended to be higher in Parkinson's disease tissue. We think that this is important preliminary evidence that diorthosubstituted PCBs may contribute to the pathogenesis of Parkinson's disease, and a greater presence of organochlorine insecticides in the PD tissue suggests that this may be in part the explanation for the association between PD and rural living.

Mitigating the effects of halofuginone on skin strength by feeding L-proline to broiler chickens.

Two experiments were conducted to determine whether supplemental levels of L-proline in the diets of broiler chickens would mitigate the skin weakening effect caused by continuous feeding of the anticoccidial halofuginone. In Experiment 1, tensile strength and collagen levels in thigh apteria skin were determined at 21 and 42 d of age in male broilers fed 0, .5, and 1% L-proline with either halofuginone (3 mg/kg) or salinomycin (61 mg/kg). In Experiment 2, the same measurements were made on female broilers receiving diets containing halofuginone and supplemented with 0, .5, or 1% L-proline, 1% L-proline through 21 d of age, or 1% L-glutamic acid through 21 d of age, or a diet containing high L-proline feedstuffs (corn gluten meal and ring dried blood meal). In Experiment 1, dermis thickness of thigh apteria was measured in the males at Day 21. Skin strength was increased in male and female broilers fed halofuginone with addition of .5 and 1% L-proline, respectively, at 21 and 42 d of age. Continuous incorporation of synthetic L-proline into diets was shown to improve skin strength in females, whereas diets formulated to contain high levels of L-proline from feedstuffs, 21-d feeding of L-proline, or L-glutamic acid did not increase skin strength.

Dietary and environmental factors affecting skin strength in broiler chickens.

Two experiments were conducted to evaluate dietary and environmental factors involved in skin tensile strength of commercial broilers. In Experiment 1 the effect of added dietary fat (4 or 7%), environmental temperature (25 or 20.5 C after 21 d), and anticoccidial drug (halofuginone or salinomycin fed continuously) were examined factorially using male and female chicks. Skin tensile strength was measured at 21, 35, and 40 d of age. Thickness of the dermal layers was measured from skin taken at Day 35. In Experiment 2, the effect of added dietary fat (0 or 7%), environmental temperature (25 or 18.5 C after 21 d), and anticoccidial drug (halofuginone or salinomycin) were examined factorially using female chicks. Skin strength and collagen content of the skin were measured at 21, 38, and 42 d of age. Skin tensile strength increased with age in both experiments, but female skin strength was subject to periodic decline. Males had significantly strong skin than females. Levels of added fat or environmental temperature did not affect skin strength in either experiment. Continuous feeding of halofuginone significantly (P < .0001) decreased skin strength compared with that of birds fed salinomycin in both experiments. Halofuginone reduced skin strength in females more than males (25 and 9%, respectively). Dermis thickness was correspondingly reduced in the birds consuming halofuginone. In Experiment 2, soluble collagen contents were reduced at all ages in birds consuming halofuginone; insoluble collagen was significantly decreased at 21 d of age. Birds with weakened skin exhibited increased incidence of skin tears during slaughter in a commercial processing plant (P < or = .0043). These results suggest that halofuginone interferes with collagen synthesis, causing decreased collagen formation and reduced skin strength. Neither added dietary fat nor ambient temperature were involved.

Effect of halofuginone on broiler skin strength when used in a shuttle anticoccidial program.

Continuous feeding of the anticoccidial halofuginone to broilers is associated with reduced skin tensile strength and increased skin tearing during processing. The possible mitigating effect of shuttle administration of halofuginone and salinomycin to female broilers was evaluated. Halofuginone or salinomycin were included in the starter and grower diets in all four possible combinations, with anticoccidial omitted from the finisher diets. Starter, grower, and finisher diets were fed to broilers through 3, 6, and 7 wk of age, respectively. Skin strength of pullets fed a diet based on milo and corn (NW) vs a diet based on corn was also compared in a factorial arrangement. Two further treatments were also included: 1) halofuginone-only NW diet supplemented with 2,500 ppm ascorbic acid from 0 to 7 wk; and 2) NW diet reared on wire floor without anticoccidial treatment. Skin tensile strength was determined at 3, 6, and 7 wk of age. Dietary composition had no effect upon skin strength or BW of broilers. Addition of ascorbic acid to the diet containing halofuginone anticoccidial did not improve skin strength. Continuous feeding of halofuginone reduced skin strength whereas withholding anticoccidial and continuous feeding of salinomycin resulted in high skin strength. When halofuginone was used in shuttle feeding programs with salinomycin, there were no differences in skin strength at 7 wk of age compared to birds that were continuously treated with salinomycin. These results suggest halofuginone may be used in a shuttle program either during the starter or grower phase without adverse affect on skin tensile strength at slaughter.

Research note: effect of electronic treatment of drinking water on growth performance of broiler chickens.

Three electronic devices used to treat drinking water were compared with untreated city well water in a broiler growth performance trial. In each treatment feed conversion and BW were measured in 16 replicate pens of 60 female broilers at 29, 42, and 49 days of age. Two of the devices increased dissolved oxygen content of the water and reduced conductivity and microorganism count. One of these reduced broiler mortality (P = .065), decreased pH (P = .062), and increased Fe and Mn concentration in the water. The other increased water temperature, Ti, and Mn but reduced the concentration of Cl, Al, Ca, Cr, Mg, and Sr in the water. Neither the third device, an electrostatic water treatment, nor the other devices affected BW (P = .586) or feed conversion (P = .564) at 49 days of age. No significant treatment differences (P less than or equal to .05) in hematocrit, bursa of Fabricius weight, or tibial ash weight were observed at 21 days of age.

Effect of cimetidine on eggshell quality and plasma 25-hydroxycholecalciferol in laying hens.

Experiments were conducted to investigate the effect of feeding cimetidine (CIMET), ranging from 0 to 750 mg/kg, on vitamin D3 metabolism and eggshell calcification in laying hens fed two levels of vitamin D3 (500 and 2,000 ICU/kg). Final BW and feed intake were not significantly affected by either CIMET or vitamin D3 level. Feeding 500 and 750 mg of CIMET significantly decreased total egg production in hens fed either level of vitamin D3, but no differences were observed at lower CIMET levels. Tibia ash decreased significantly in hens fed 150 to 750 mg of CIMET, regardless of the vitamin D3 level. Plasma Ca and inorganic P concentrations were decreased in hens fed high CIMET levels (500 and 750 mg/kg) at Week 2, but no differences were observed at Week 4. Feeding CIMET (500 and 750 mg/kg) significantly decreased plasma 25-hydroxycholecalciferol (25-OHD3) levels at Week 2 in hens fed both vitamin D3 diets but not at Week 4. Eggshell breaking force, shell thickness, and percentage shell weight were decreased significantly by CIMET in all experiments; however, in one experiment, shell quality recovered by Week 8. These results suggest that the CIMET-induced reduction in bone mineralization, eggshell quality, and plasma 25-OHD3 levels could be due to interference of CIMET with vitamin D3 metabolism in vitamin D3-replete laying hens. Shell quality decreased in CIMET-treated hens fed the higher vitamin D diet even though 250-HD3 plasma levels were three times higher than in hens fed the lower vitamin D diet, suggesting that CIMET affected shell quality through some mechanism other than inhibition of 250-HD3 synthesis.

Effect of feeding isoniazid and cimetidine on growth and bone development in male broiler chicks.

Because previous studies indicated that cimetidine (CIMET) and isoniazid (ISON) may inhibit vitamin D metabolism in both rats and humans, experiments were conducted to evaluate the influence of these drugs on growth and bone development in male broiler chicks. Chicks were fed a corn and soybean meal basal diet containing various levels of the drugs for 21 days. Body weight gain (BWG), feed consumption per bird (FB), tibia ash (TA), tibia breaking force (TBF), and plasma calcium levels were measured. In birds fed a diet supplemented with 1,100 ICU D3/kg, CIMET at up to 300 ppm resulted in a significant linear decrease in TA as the drug level increased. Birds fed ISON up to 405 ppm had inconsistent responses in TA and TBF. Body weight gain was only depressed with the highest level of ISON (405 ppm). The effects of feeding 0, 150, or 300 ppm CIMET were also investigated when chicks were fed 200 or 1,000 ICU per kg vitamin D3. The interaction between vitamin D3 and CIMET was significant for BWG, FCB, TA, and TBF. These were significantly reduced as the CIMET level increased for birds fed the low vitamin D3 diet but were not significantly affected when fed the high D3 diet. In another study in which chicks were fed a diet with 1,100 ICU vitamin D3, lowering the dietary Ca, or P, or both did not result in any effect of CIMET on the chicks. The results indicate that CIMET possibly interferes with normal bone formation in chicks by altering vitamin D3 metabolism.

Sulfur amino acid requirement of broiler chickens from 3 to 6 weeks of age.

Two experiments were conducted to determine if the TSAA level suggested by the National Research Council (NRC) in 1984 was adequate for optimum performance of 3 to 6-wk-old broilers, and to assess the effect of added copper on the TSAA requirement. A 2 x 3 x 2 factorial arrangement involving two copper (0 and 240 ppm) and three TSAA levels (.72, .78 and .84%) and two sexes was used in each experiment. Copper sulfate (CuSO4.5H2O) and DL-methionine were the dietary supplements to pelleted diets. There were four groups of 20 males or 20 females in each experiment, housed in floor pens. Methionine supplementation significantly improved body weight gain and feed efficiency in males but not in females. A significant copper x methionine interaction was observed, with methionine supplementation improving feed efficiency more in the presence of copper than in its absence. Abdominal fat per unit of body weight was significantly reduced by both methionine and copper in females but not in males. In a second experiment, methionine significantly improved body weight gain in males fed no added copper and significantly reduced abdominal fat per unit of body weight. Methionine supplementation improved body weight gain in males more in the absence than in the presence of copper (significant TSAA x copper interaction). Methionine supplementation had no significant effects on performance of females in the second experiment, but copper supplementation significantly improved feed efficiency.(ABSTRACT TRUNCATED AT 250 WORDS)

CIRRIS-1A interferometer: radiometric analysis.

The CIRRIS-lA spectroradiometer system is designed to obtain spectral and spatial airglow data on the shuttle platform over a global extent. The sensor system includes a Michelson interferometer which exhibits a noise equivalent sterance [radiance] (NER) of 2 x 10(-13) W cm(-2) sr(-1) Hz(-1/2); and at a resolution of 0.964 cm(-1) (scan time 9.1 s) exhibits a noise equivalent spectral sterance [radiance] NESR of 7 x 10(-14) W cm(-2)sr(-1)/cm(-1). The entire optical subsystem, silicon-arsenic focal-plane, off-axis high-rejection telescope, and removable cover are cooled below 20 K. The measured performance is shown to agree with theory to within a few percent. Noise-equivalent-sterance [radiance] per root-Hz is recommended as a figure of merit for interferometers that is independent of the specific operating conditions; i.e., mirror velocity, scan time or transform size.

Influence of hatcher holding times on several physiological parameters associated with the immune system of chickens.

Two experiments were conducted with broiler chicks to study the effects of posthatch holding time in the incubator on physiological and disease parameters. In each experiment, half of the chicks were removed shortly after hatching and half of the chicks were allowed to remain in the hatcher for an additional 30 hr. Bursa weights, hematocrits, total plasma protein, and blood glucose concentrations were measured at various times up to 35 and 28 days of age in Experiments 1 and 2, respectively. In addition, chicks hatched from eggs with one of two specific gravities (less than or equal to 1.065 and greater than or equal to 1.075) were tested in the second experiment. Spleen weights, heterophil:lymphocyte ratios and the influence of an aerosol Newcastle disease-Mycoplasma gallisepticum vaccine challenge were also measured in the second experiment. In Experiment 1, when chicks were held in the hatcher for an additional 30 hr, bursa weights were reduced through 8 days of age; however, they were significantly heavier by 21 days of age. In Experiment 2, both bursa and spleen weights were significantly reduced through 14 days of age when chicks were held in the incubator. Total plasma protein and glucose concentrations, hematocrits, and heterophil:lymphocyte ratios were all significantly higher for chicks held in the incubator. Chicks held in the incubator and then exposed to an aerosol vaccine challenge at 1 day of age had a significantly greater percentage of air sac lesions and the lesions were more severe at 28 days of age. Egg specific gravity had no influence on any of the parameters measured.(ABSTRACT TRUNCATED AT 250 WORDS)

Infrared spectrometer: liquid-helium-cooled rocketborne circular-variable filter.

An ir spectrometer used to obtain atmospheric emission spectra from a rocket at altitudes between 45 km and 200 km in the ICECAP Auroral Measurements Program is described. The instrument continuously scans the 6.75-23.2-microm spectral region at the rate of 2 scans/sec. The spectral resolution of the instrument, which employs a circular-variable interference filter (CVF), ranges from 3% to 4%. The entire optical subsection, silicon-arsenic detector, CVF, Irtran 6 lens, baffle, and removable cold cover are cooled below 10 K in a high-vacuum Dewar system. The noise equivalent spectral radiance (NESR) of the spectrometer at 22 ,microm is better than 1 x 10(-11) (W cm(-2)sr(-1)microm(-1)). The dominant high altitude atmospheric emission features observed on two rocket flights were 9.6-microm O(3) and 15-microm CO(2).

Multiplexed dispersive spectrometers using reduced background infrared detectors.

The application of multiplex spectrometry to cryogenically cooled LWIR extrinsic photodetectors is limited by system noise. This noise limitation results in a detector NEP that is directly proportional to bandwidth. Therefore, multiplex schemes that require increased bandwidth are not productive of real advantage. However, doubly encoded systems that are based on 2n - 1 or n + N - 1 measurements have the potential to provide a real throughput gain proportional to the number of elements used on the throughput matrix.