Investigation of connexin 43 uncoupling and prolongation of the cardiac QRS complex in preclinical and marketed drugs.

BACKGROUND AND PURPOSE: Prolongation of the cardiac QRS complex is linked to increased mortality and may result from drug-induced inhibition of cardiac sodium channels (hNaV 1.5). There has been no systematic evaluation of preclinical and marketed drugs for their additional potential to cause QRS prolongation via gap junction uncoupling. EXPERIMENTAL APPROACH: Using the human cardiac gap junction connexin 43 (hCx43), a dye transfer 'parachute' assay to determine IC50 values for compound ranking was validated with compounds known to uncouple gap junctions. Uncoupling activity (and hNaV 1.5 inhibition by automated patch clamp) was determined in a set of marketed drugs and preclinical candidate drugs, each with information regarding propensity to prolong QRS. KEY RESULTS: The potency of known gap junction uncouplers to uncouple hCx43 was ranked (according to IC50 ) as phorbol ester>digoxin>meclofenamic acid>carbenoxolone>heptanol. Among the drugs associated with QRS prolongation, 29% were found to uncouple hCx43 (IC50 < 50 µM), whereas no uncoupling activity was observed in drugs not associated with QRS prolongation. In preclinical candidate drugs, hCx43 and hNaV 1.5 IC50 values were similar (within threefold). No consistent margin over preclinical Cmax (free) was apparent for QRS prolongation associated with Cx43 inhibition. However, instances were found of QRS prolonging compounds that uncoupled hCx43 with significantly less activity at hNaV 1.5. CONCLUSION AND IMPLICATIONS: These results demonstrate that off-target uncoupling activity is apparent in drug and drug-like molecules. Although the full ramifications of Cx inhibition remain to be established, screening for hCx43 off-target activity could reduce the likelihood of developing candidate drugs with a risk of causing QRS prolongation.

Recovery of human motoneurons during rotation.

During prolonged contractions, few studies have reported rotation among low threshold motoneurons. The question arises whether a motoneuron stops firing due to an increase in firing threshold or whether it is due to regional switching of activity among muscle fascicles. We postulated that if the rest period resulted from an increase in firing threshold, a progressive recovery in the excitability of the motoneuron would be observed during the rest period. The excitability of soleus or tibialis anterior motoneurons was tested during the rest periods. The results showed that a previously tonic motoneuron that had dropped off during rotation, rarely responded to Ia or TMS inputs in the initial parts of the rest period; however, its response probability increased significantly in the second half. Based on these data, we suggest that the observed rotation is due to changes in firing thresholds of motoneurons during prolonged firing.

Influence of supplemental dietary poultry fat on the digestive and reproductive organ characteristics of commercial layers inoculated before or at the onset of lay with F-strain Mycoplasma gallisepticum.

Effects of F-strain Mycoplasma gallisepticum (FMG) inoculation and 1.5% supplemental dietary poultry fat (PF) on the digestive and reproductive organ characteristics of commercial layers at 58 wk of age were investigated. Sham and FMG inoculations were administered at 12 (before lay) and 22 (early in lay) wk of age, and dietary treatments (basal control diets and basal control diets with PF) were initiated at 20 wk of age. Supplemental PF increased BW and decreased isthmal length relative to total oviduct length in hens. Various oviduct segments were also affected by the type and age of inoculation, and these effects were further influenced by the use of PF. In comparison to their time-specific sham-inoculated controls, infundibulum weight relative to BW was increased when birds were inoculated with FMG at 22 wk, whereas isthmus weight relative to total oviduct weight was increased by FMG inoculation at 12 wk of age. However, PF affected infundibulum length relative to total oviduct length only in sham-inoculated birds, and PF increased magnum weight relative to total oviduct weight only in birds inoculated at 22 wk of age (sham or FMG). Furthermore, PF decreased isthmus weight relative to total oviduct weight only in birds that were sham-inoculated (12 or 22 wk). In conclusion, the inoculation of FMG at 12 or 22 wk may increase the relative contributions of the isthmus and infundibulum, respectively, to the total mass of the oviduct. In addition, PF may decrease the relative length of the isthmus and increase the relative weight of the magnum in the oviducts of birds that have been inoculated at 22 wk of age (sham or FMG). Previous studies have shown 1.5% supplemental dietary PF to influence feed consumption throughout lay and performance early in lay in hens that were inoculated with FMG at 12 wk of age. However, the current results suggest that these influences are associated with gross changes in the oviduct but not the digestive tract of layers.

Influence of supplemental dietary poultry fat on the yolk characteristics of commercial layers inoculated before or at the onset of lay with F-strain Mycoplasma gallisepticum.

Effects of F-strain Mycoplasma gallisepticum (FMG) inoculation and 1.5% supplemental dietary poultry fat (PF) on the egg yolk characteristics of commercial layers at 24, 34, 44, 50, and 58 wk of age were investigated. Sham and FMG inoculations were administered at 12 and 22 wk of age and dietary treatments (basal control and basal control with 1.5% supplemental PF) were initiated at 20 wk of age. Yolk lipid concentration was reduced on wk 24 in birds that had been inoculated at 12 or 22 wk of age with FMG. The use of 1.5% supplemental PF increased percentage of yolk weight and yolk:albumen ratio across age and inoculation treatment. At 58 wk of age, concentrations of yolk palmitic acid increased and those of oleic and linolenic acid decreased when sham inoculations were given at 22 rather than at 12 wk of age. However, FMG inoculations given at 22 rather than at 12 wk increased palmitoleic acid and decreased stearic acid yolk concentrations. At 12 wk of age, FMG inoculations decreased yolk palmitoleic, oleic, and linolenic acid concentrations while causing increased yolk stearic and arachidonic acid levels when compared with sham inoculations. Furthermore, 1.5% supplemental PF decreased concentrations of palmitic and oleic acid and increased those of linoleic acid in the yolk at 58 wk of age. Despite the interaction of 1.5% supplemental PF with the prelay inoculation of FMG on early (18 to 26 wk) layer performance noted in a previous report, the effects of a prelay FMG inoculation and 1.5% supplemental PF on the egg yolk characteristics examined in the current study were independent of each other. This suggests that 1.5% supplemental PF is not effective in modulating the effects of an FMG inoculation at 12 wk of age on hen egg yolk characteristics between 24 and 58 wk of age and that the combined effects of PF supplementation and FMG inoculation on performance do not influence egg yolk characteristics.

Influence of supplemental dietary poultry fat on the performance of commercial layers inoculated before or at the onset of lay with F-strain Mycoplasma gallisepticum.

The effects of F-strain Mycoplasma gallisepticum (FMG) inoculation and 1.5% supplemental dietary poultry fat (PF) on the performance of commercial layers between 20 and 58 wk of age were investigated. Sham and FMG inoculations were administered at 12 (before lay) and 22 (early in lay) wk and dietary treatments (basal control diets and basal control diets with 1.5% supplemental PF) were initiated at 20 wk of age. Mortality at wk 47 and 53 was greatest in birds inoculated with FMG at 22 wk. Feed consumption from 20 to 23 and from 52 to 55 wk of age was greater in birds that were inoculated with FMG (12 or 22 wk). However, feed consumption decreased in birds that were inoculated at 12 wk (sham or FMG) when 1.5% supplemental PF was added to the diet. Percentage of total egg production (EP) between 22 and 58 wk of age was highest in hens that were inoculated with FMG at 22 wk. Furthermore, weekly EP increased at wk 27 and 58 and decreased at wk 47 after birds had been inoculated with FMG (12 or 22 wk), and increased at 22 wk and decreased at 54 wk when inoculations (sham or FMG) were given at 22 wk. Egg weight was increased at wk 29, 31, 39, 40, 42, 44, 53, and 58 in birds that were inoculated with FMG (12 or 22 wk); however, there were no coherent treatment effects on eggshell quality. An FMG inoculation at 22 wk may promote total EP through 58 wk, whereas the inoculation of commercial layers with FMG (12 or 22 wk) may increase subsequent feed consumption during the early and late stages of EP and increase egg weight throughout lay. However, the supplementation of hen diets with 1.5% PF beginning at 20 wk of age may reduce subsequent feed consumption throughout lay in birds having experienced a prelay (12 wk) inoculation.

Evaluation of an experimental chlorate product as a preslaughter feed supplement to reduce salmonella in meat-producing birds.

A study was conducted to evaluate the effect of experimental chlorate product (ECP) feed supplementation on Salmonella Typhimurium (ST) in the crop and ceca of market-age broilers. In trial 1, 160 market-age broilers were randomly assigned to 8 treatment groups and replicated twice, with 20 broilers per pen for 1 wk. Trial 2 used the same design, but used 80 market-age broilers with 10 broilers per pen. Treatments were as follows: 1) control feed + double-distilled drinking water (dd H(2)O); 2) control + 18.5% experimental zeolite carrier with dd H(2)O; 3 to 7) control feed supplemented with 0.5, 1.0, 5.0, 10.0, or 18.5% of a feed grade ECP + dd H(2)O; 8) control feed + 1x ECP (0.16% w/v; containing 15 mM chlorate ion equivalent) added to dd H(2)O. Seven-week-old broilers were provided experimental treatments for 7 d, killed, and then ceca and crops were removed and evaluated for ST. Broilers fed 5 to 18.5% ECP or water ECP had a significantly lower (P < 0.05) incidence of ST in the crop (36 to 38% and 14%, respectively) when compared with the control (60%). Broilers fed 10% ECP or water ECP had significantly lower ST crop concentrations (1.03 log(10) and 0.38 log(10) ST/g, respectively) when compared with broilers fed a control diet (1.54 log(10) ST/g). Crop and ceca ST incidence (32 to 48%) and concentration (1.00 to 1.82 log(10) ST/g) were significantly lower in broilers fed 5 to 18.5% ECP as compared with the control (78%; 2.84 log(10) ST/g). Broilers fed 5% or greater ECP had significantly higher water consumption (380 to 580 mL water/d) and litter moisture (31 to 56%) when compared with the control (370 mL water/d; 23% moisture). Only broilers fed 18.5% ECP had significantly lower 7-wk BW (2.77 kg of BW) when compared with the controls (3.09 kg of BW). Average daily gains were significantly depressed in broilers fed 10 or 18.5% ECP compared with the controls. These results indicate broilers supplemented with feed

Effects of supplemental dietary phytase and 25-hydroxycholecalciferol on the performance characteristics of commercial layers inoculated before or at the onset of lay with the F-strain of Mycoplasma gallisepticum.

The effects of dietary supplementation with phytase and 25-hydroxycholecalciferol on the performance characteristics of commercial layers that were inoculated prelay (12 wk of age) or at the onset of lay (22 wk of age) with F-strain Mycoplasma gallisepticum were assessed. Experimental layer diets, which included a basal control diet or the same diet supplemented with 0.025% phytase and 25-hydroxycholecalciferol, were fed from 20 through 58 wk of age. Weekly and total egg production were determined from 22 through 58 wk, and egg weight and various internal egg and eggshell quality characteristics were examined at 34, 50, and 58 wk of age. F-strain M. gallisepticum inoculation decreased egg production at the beginning of lay (wk 22 and 23) but increased post-peak lay at wk 45. However, there were no treatment effects of any kind on total egg production, egg weight, or any of the internal egg and eggshell characteristics examined during lay. In conclusion, dietary supplementation with phytase and 25-hydroxycholecalciferol did not affect layer performance or interact with the effects of F-strain M. gallisepticum inoculation; however, F-strain M. gallisepticum inoculation resulted in a shift in egg production from wk 22 to 45 without having an overall effect on total egg production.

Effects of supplemental dietary phytase and 25-hydroxycholecalciferol on the digestive and reproductive organ characteristics of commercial layers inoculated before or at the onset of lay with the F-strain of Mycoplasma gallisepticum.

In 3 trials, the effects of dietary supplementation with phytase (PHY) and 25-hydroxycholecalciferol (25-D3) on the digestive and reproductive organ characteristics of commercial layers that were inoculated prelay (12 wk of age) or at the onset of lay (22 wk of age) with F-strain Mycoplasma gallisepticum (FMG) were assessed at 58 wk of age. Experimental layer diets that included a basal control diet or a control diet supplemented with 0.025% PHY and 25-D3 were fed from 20 through 58 wk of age. As a percentage of total oviduct weight, magnum weight was lower in birds that were inoculated (sham or FMG) at lay onset compared with those that were inoculated prelay, and in FMG-inoculated birds, relative duodenum length was greater in those inoculated at 12 compared with 22 wk. Also, as percentages of organ weight or length, infundibulum length and isthmus weight were increased, whereas duodenum length was decreased by dietary supplementation with PHY and 25-D3. The overall timing (12 vs. 22 wk) of inoculation can affect the reproductive organ characteristics of layers, whereas, more specifically, the timing of an FMG inoculation may affect their digestive organ structure. Furthermore, independent of inoculation timing and type, the reproductive organ and digestive systems of laying hens may be influenced by dietary supplementation with PHY and 25-D3.

Effects of methyl beta-cyclodextrin on EDHF responses in pig and rat arteries; association between SK(Ca) channels and caveolin-rich domains.

BACKGROUND AND PURPOSE: The small and intermediate conductance, Ca2+-sensitive K+ channels (SK(Ca) and IK(Ca), respectively) which are pivotal in the EDHF pathway may be differentially activated. The importance of caveolae in the functioning of IK(Ca) and SK(Ca) channels was investigated. EXPERIMENTAL APPROACH: The effect of the caveolae-disrupting agent methyl-beta-cyclodextrin (MbetaCD) on IK(Ca) and SK(Ca) localization and function was determined. KEY RESULTS: EDHF-mediated, SK(Ca)-dependent myocyte hyperpolarizations evoked by acetylcholine in rat mesenteric arteries (following blockade of IK(Ca) with TRAM-34) were inhibited by MbetaCD. Hyperpolarizations evoked by direct SK(Ca) channel activation (using NS309 in the presence of TRAM-34) were also inhibited by MbetaCD, an effect reversed by cholesterol. In contrast, IK(Ca)-dependent hyperpolarizations (in the presence of apamin) were unaffected by MbetaCD. Similarly, in porcine coronary arteries, EDHF-mediated, SK(Ca)-dependent (but not IK(Ca)-dependent) endothelial cell hyperpolarizations evoked by substance P were inhibited by MbetaCD. In mesenteric artery homogenates subjected to sucrose-density centrifugation, caveolin-1 and SK3 (SK(Ca)) proteins but not IK1 (IK(Ca)) protein migrated to the buoyant, caveolin-rich fraction. MbetaCD pretreatment redistributed caveolin-1 and SK3 proteins into more dense fractions. In immunofluorescence images of porcine coronary artery endothelium, SK3 (but not IK1) and caveolin-1 were co-localized. Furthermore, caveolin-1 immunoprecipitates prepared from native porcine coronary artery endothelium contained SK3 but not IK1 protein. CONCLUSIONS AND IMPLICATIONS: These data provide strong evidence that endothelial cell SK(Ca) channels are located in caveolae while the IK(Ca) channels reside in a different membrane compartment. These studies reveal cellular organisation as a further complexity in the EDHF pathway signalling cascade.

Effects of supplemental dietary phytase and 25-hydroxycholecalciferol on the blood characteristics of commercial layers inoculated before or at the onset of lay with the F-strain of Mycoplasma gallisepticum.

In 3 trials, the effects of dietary supplementation with phytase (PHY) and 25-hydroxycholecalciferol on BW and the blood characteristics of commercial layers that were inoculated prelay (12 wk of age) or at the onset of lay (22 wk of age) with F-strain Mycoplasma gallisepticum were assessed at 34, 50, and 58 wk of age. Experimental layer diets, which included either a basal control diet or the same diet supplemented with 0.025% PHY and 25-hydroxycholecalciferol, were fed from 20 through 58 wk of age. The supplemented diet decreased blood hematocrit values across bird age, inoculation type (sham vs. F-strain M. gallisepticum), and age of inoculation (prelay vs. onset of lay). Phytase- and 25-hydroxycholecalciferol-supplemented diets reduced bird BW in sham-inoculated control birds across bird age and age of inoculation. This effect was not observed in F-strain M. gallisepticum-inoculated birds. Furthermore, across diet (control vs. supplemented) and inoculation type, total plasma protein concentration at 34 wk of age was higher in birds that were inoculated at the onset of lay compared with those inoculated prelay. Diet, inoculation type, and inoculation age had no effect on mortality, reproductive organ histopathological lesion scores, or serum cholesterol and Ca concentrations. In conclusion, throughout lay, the supplementation of commercial layer diets with PHY may lower hematocrit, and inoculation with F-strain M. gallisepticum prelay or at the onset of lay may ameliorate the depressing effects of dietary PHY and 25-hydroxycholecalciferol supplementation on hen BW.

Comparison of zinc acetate and propionate addition on gastrointestinal tract fermentation and susceptibility of laying hens to Salmonella enteritidis during forced molt.

Feed deprivation is the most common method used to induce molting and stimulate multiple egg-laying cycles in laying hens for commercial egg production. Unfortunately, an increased risk of Salmonella enteritidis (SE) colonization may result from the use of this method. Methods to stimulate multiple egg-laying cycles without increasing the risk of SE are needed. In each of 3 experiments, hens over 50 wk of age were divided into groups of 12 and placed in individual laying cages. One week before dietary changes, hens were put on an 8L:16D photoperiod that continued for the 9-d experimental period. Hens in all treatments were challenged orally with 10(4) cfu of SE on the fourth day. Treatments were full fed hens (nonmolted, NM), nonfed hens (molted, M), a zinc acetate diet (ZAC), and a zinc propionate diet (ZPR). The zinc diets contained 10,000 mg of zinc per kilogram of diet. Body weight losses were significantly higher in the M, ZPR, and ZAC treatments than in the NM treatment. Crop lactic acid decreased more in M, ZPR, and ZAC treatments than in NM hens in trial 2. Crop pH was significantly (P < 0.05) lower in NM hens than in M, ZAC, and ZPR hens in trial 2. Although cecal individual or total volatile fatty acids (VFA), and lactic acid were not significantly (P > 0.05) different between NM hens and M, ZAC and ZPR hens in trial 1, lactic acid was significantly (P < 0.05) higher in NM hens than in M, ZAC and ZPR hens (trial 2), and cecal total VFA were lower in M hens than in NM, ZAC and ZPR hens (trial 3). Colonization of SE in the crop and ceca was higher in the M and ZPR hens (trials 1 and 2). Liver, spleen, or ovary invasion by SE was higher in the M and ZPR hens (trials 1 and 2) than in NM hens. At the zinc concentration used in these studies, the zinc dietary regimens may be effective for reducing the risk of SE during induced molt.

Effects of fasting on serum lipids and lipoprotein profiles in the egg-laying hen (Gallus domesticus).

The effects of a 24-h fast on serum lipids and lipoprotein profiles in commercial laying hens were investigated. Blood was analyzed at 34 and 46 weeks of age from Single Comb White Leghorn hens that had been either fed ad libitum or had been fasted for 24 h prior to collection. At 12 weeks, birds were divided into 16 biological isolation units, with 8 replicate units assigned to each treatment group. Four birds out of 10 in each unit were tagged for bleeding. Parameters evaluated included total serum cholesterol and triglycerides, mean diameters of very low density lipoproteins (VLDLs) for the 10th, 50th, and 90th percentiles of serum total VLDL, mean total population VLDL particle diameter (MPD), and percentage serum cholesterol recovered in VLDL, low density lipoprotein (LDL), and high density lipoprotein (HDL) fractions. Fasting led to decreases in total serum cholesterol and triglycerides, and a decrease in mean serum VLDL particle diameter in the 90th population percentile. At Week 34, percentage serum cholesterol recovered from LDL was increased, whereas percentage serum cholesterol recovered from HDL was decreased due to fasting. At Week 46, MPD and percentage serum cholesterol recovered from VLDL were decreased, whereas percentage serum cholesterol recovered from HDL was increased due to fasting. It was concluded that a 24-h fast decreased serum lipids (cholesterol and triglycerides) and the size of VLDL particles in the 90th population percentile in commercial laying hens. Furthermore, bird age influenced the effects of a 24-h fast on MPD and the redistribution of serum cholesterol among VLDL, LDL, and HDL particles.

Functional evidence of a role for two-pore domain potassium channels in rat mesenteric and pulmonary arteries.

1. Experiments were performed to elucidate the mechanism by which alterations of extracellular pH (pH(o)) change membrane potential (E(M)) in rat mesenteric and pulmonary arteries. 2. Changing pH(o) from 7.4 to 6.4 or 8.4 produced a depolarisation or hyperpolarisation, respectively, in mesenteric and pulmonary arteries. Anandamide (10 microm) or bupivacaine (100 microm) reversed the hyperpolarisation associated with alkaline pH(o), shifting the E(M) of both vessels to levels comparable to that at pH 6.4. In pulmonary arteries, clofilium (100 microm) caused a significant reversal of hyperpolarisation seen at pH 8.4 but was without effect at pH 7.4. 3. K(+) channel blockade by 4-aminopyridine (4-AP) (5 mm), tetraethylammonium (TEA) (10 mm), Ba(2+) (30 microm) and glibenclamide (10 microm) depolarised the pulmonary artery. However, shifts in E(M) with changes in pH(o) remained and were sensitive to anandamide (10 microm), bupivacaine (100 microm) or Zn(2+) (200 microm). 4. Anandamide (0.3-60 microm) or bupivacaine (0.3-300 microm) caused a concentration-dependent increase in basal tone in pulmonary arteries. 5. RT-PCR demonstrated the expression of TASK-1, TASK-2, THIK-1, TRAAK, TREK-1, TWIK-1 and TWIK-2 in mesenteric arteries and TASK-1, TASK-2, THIK-1, TREK-2 and TWIK-2 in pulmonary arteries. TASK-1, TASK-2, TREK-1 and TWIK-2 protein was demonstrated in both arteries by immunostaining. 6. These experiments provide evidence for the presence of two-pore domain K(+) channels in rat mesenteric and pulmonary arteries. Collectively, they strongly suggest that modulation of TASK-1 channels is most likely to have mediated the pH-induced changes in membrane potential observed in these vessels, and that blockade of these channels by anandamide or bupivacaine generates a small increase in pulmonary artery tone.

Effects of F-strain Mycoplasma gallisepticum inoculation on serum very low density lipoprotein diameter and fractionation of cholesterol among lipoproteins in commercial egg-laying hens.

Experimental inoculation with the F-strain of Mycoplasma gallisepticum (FMG) at 12 wk of age has been shown to affect the performance, liver, reproductive organs, and yolk lipid characteristics of commercial layers. Therefore, this study was conducted to determine the serum lipoprotein characteristics of commercial egg-laying hens at 16 wk of age and throughout lay after inoculation with FMG at 12 wk of age. Mean diameters of very low density lipoproteins (VLDL) were determined for the 10th, 50th, and 90th percentiles of serum total VLDL of each hen. Percentages of total serum cholesterol recovered in VLDL and low and high density lipoprotein particle classes were also determined. Inoculation of birds with FMG at 12 wk did not change the physical properties or relative concentrations of their circulating lipoproteins. However, the age of the bird had significant differential effects on all the parameters examined. These data demonstrate that FMG-inoculation at 12 wk of age does not affect the lipoproteins of laying hens, but because these birds were housed in biological isolation units, these results do not preclude the possibility that these yolk precursors may be affected in FMG-infected birds that are housed in facilities in which there are increased levels of environmental stress. These data further suggest that alterations in liver, reproductive organs, and yolk lipid characteristics in response to FMG, as noted in previous reports on commercial layers, are not mediated through changes in circulating VLDL diameters.

Effects of F-strain Mycoplasma gallisepticum inoculation at twelve weeks of age on the blood characteristics of commercial egg laying hens.

In two trials, the effects of an F-strain Mycoplasma gallisepticum (FMG) inoculation at 12 wk of age on the blood characteristics of commercial Single Combed White Leghorn laying hens were investigated throughout lay. Variables measured in both trials were whole blood hematocrit, plasma protein (PP), and serum cholesterol, triglycerides (ST), and calcium. In both trials, hematocrit at 20 wk of age was significantly increased in birds inoculated with FMG. In trial 1, ST and PP were significantly increased at 22 wk of age by FMG, while ST and PP were significantly decreased in FMG-inoculated birds at wk 54 and 52, respectively. When combined with the establishment of an FMG infection, the initial weeks of egg production become particularly stressful to the bird. Increases in these independent blood parameters between 8 and 10 wk postchallenge are suggestive of compensatory responses in these birds to an FMG challenge. Postpeak decreases in both ST (54 wk) and PP (52 wk) in FMG-infected birds may be the result of a more chronic effect of FMG on lipid and protein synthesis in the liver. These data are the first to suggest that alterations in egg production in response to FMG-infection in commercial layers, as noted in a previous report, may be associated with changes in hematocrit. However, because ST and PP were not affected by FMG in both trials, the responses of these blood parameters to FMG-infection may be inconsistent among flocks.

Effects of F-strain Mycoplasma gallisepticum inoculation at twelve weeks of age on egg yolk composition in commercial egg laying hens.

In two trials, the effects of F-strain Mycoplasma gallisepticum (FMG) on the contents of egg yolks from commercial Single Comb White Leghorn laying hens were investigated over a production cycle. Ten hens were assigned to each of 8 (trial 1) or 16 (trial 2) negative pressure fiberglass biological isolation units. Birds in half of the total units served as sham-inoculated controls, and those in the other half were inoculated with FMG at 12 wk of age. Eggs were collected and yolks were harvested at various times during the prepeak, peak, and postpeak periods of both trials for constituent analysis. Yolk constituents analyzed in these trials included moisture, total lipids, cholesterol, triglycerides, phospholipids, and fatty acids. In both trials, total yolk lipid at 22 wk of age was significantly decreased in birds inoculated with FMG. In trial 1, yolk cholesterol at 28 wk was significantly decreased in FMG-inoculated birds. Yolk linoleic acid in trial 1 and yolk stearic and arachidonic acids in trial 2 were significantly increased in FMG-inoculated birds compared to FMG-free birds. In trial 2, yolk myristic, palmitoleic, and oleic acid percentages were significantly decreased in FMG-inoculated birds compared to FMG-free birds. These data suggest that alterations in egg production in commercial layers in response to an FMG infection at 12 wk of age are associated with changes in yolk composition.

Influences of supplemental dietary poultry fat and F-strain Mycoplasma gallisepticum infection on the early performance of commercial egg laying hens.

F-strain Mycoplasma gallisepticum (FMG) may alter reproductive performance in layers through its effects on lipid metabolism. Therefore, the influences of 1.5% supplemental dietary poultry fat (PF) and FMG infection on the early performance of commercial egg-laying hens were determined. Birds were either sham- or FMG-inoculated at 12 wk, and experimental diets were initiated at 20 wk of age. Body weight at 12, 20, and 24 wk, total daily egg mass, feed consumption and feed conversion at 20 and 24 wk, weekly egg weight between 19 and 26 wk, weekly egg production (EP) between 18 and 26 wk, and weekly mortality between 12 and 26 wk of age were determined. Inoculation with FMG reduced EP at 18 and 19 wk of age. Between 20 and 26 wk, FMG reduced EP in birds fed control diets, conversely, PF eliminated differences in EP between sham- and FMG-inoculated birds. Furthermore, at wk 20 and 24, birds consumed less feed when fed PF-supplemented diets than when fed control diets if they were sham-inoculated, but the difference in feed consumption between diets was ameliorated if birds were previously inoculated with FMG. These data demonstrate that the effects of a 12-wk inoculation of FMG on EP and feed consumption through 26 wk of age in commercial egg-laying chickens can be modified by 1.5% supplemental dietary P F. More specifically, PF may alleviate reductions in early EP due to FMG.

Effects of F-strain Mycoplasma gallisepticum inoculation at twelve weeks of age on performance and egg characteristics of commercial egg-laying hens.

The effects of F-strain Mycoplasma gallisepticum (FMG) inoculation during the pullet period on the subsequent performance and egg characteristics of commercial Single Combed White Leghorn hens were evaluated. In two trials, BW, feed consumption, egg production (EP), egg weight, egg size class, relative eggshell water vapor conductance, and relative percentages of eggshell, yolk and albumen weights were determined through approximately 60 wk of age. In each trial, pullets at 12 wk of age were randomly assigned to negative pressure biological isolation units. Birds in one-half of the total units were inoculated with FMG, and the other half were sham-inoculated with sterile media. In both trials, onset of lay was delayed approximately 1 wk in layers inoculated with FMG. Control birds that had not been previously inoculated with FMG laid their first egg at 18 wk of age, while birds that had been previously inoculated with FMG laid their first egg at 19 wk of age. In Trial 1, FMG-inoculated hens laid significantly fewer total eggs, which became apparent at each week after Week 42. In Trial 2, a numerical decrease in total EP occurred, and the percentage of undersized eggs laid by FMG-inoculated birds was significantly lower at 19 wk of age but was higher at 20 and 21 wk when compared to controls. Mortality was not significantly different between the treatments in either trial. These data demonstrate that when birds are housed in isolation facilities and inoculated with FMG at 12 wk of age, onset of lay is delayed. These data also suggest that FMG may lead to delays in undersize EP and decreases in total EP. However, because significant FMG effects on these parameters were observed in only one trial, additional studies may be necessary to verify these effects.

Effects of an S6 strain of Mycoplasma gallisepticum challenge before beginning of lay on various egg characteristics in commercial layers.

Mycoplasma gallisepticum (MG) is a reproductive/respiratory disease in poultry implicated in suboptimum egg production and decreased hatchability. Commercial layer hens raised in a controlled environment were inoculated with the S6 strain of MG at 10 wk of age. Egg production and selected egg and egg quality parameters were quantitated over the entire lay cycle for inoculated and control birds. The S6 inoculation had no effect on bird weight, egg production, associated egg quality parameters, or histopathologic lesion scores. This study shows that in the absence of environmental stressors a prelay S6 MG inoculation does not produce detrimental effects on layer hen performance.

K+-induced hyperpolarization in rat mesenteric artery: identification, localization and role of Na+/K+-ATPases.

1. Mechanisms underlying K(+)-induced hyperpolarizations in the presence and absence of phenylephrine were investigated in endothelium-denuded rat mesenteric arteries (for all mean values, n=4). 2. Myocyte resting membrane potential (m.p.) was -58.8+/-0.8 mV. Application of 5 mM KCl produced similar hyperpolarizations in the absence (17.6+/-0.7 mV) or presence (15.8+/-1.0 mV) of 500 nM ouabain. In the presence of ouabain +30 microM barium, hyperpolarization to 5 mM KCl was essentially abolished. 3. In the presence of 10 microM phenylephrine (m.p. -33.7+/-3 mV), repolarization to 5 mM KCl did not occur in the presence or absence of 4-aminopyridine but was restored (-26.9+/-1.8 mV) on addition of iberiotoxin (100 nM). Under these conditions the K+-induced repolarization was insensitive to barium (30 microM) but abolished by 500 nM ouabain alone. 4. In the presence of phenylephrine + iberiotoxin the hyperpolarization to 5 mM K(+) was inhibited in the additional presence of 300 nM levcromakalim, an action which was reversed by 10 microM glibenclamide. 5. RT-PCR, Western blotting and immunohistochemical techniques collectively showed the presence of alpha(1)-, alpha(2)- and alpha(3)-subunits of Na(+)/K(+)-ATPase in the myocytes. 6. In K(+)-free solution, re-introduction of K(+) (to 4.6 mM) hyperpolarized myocytes by 20.9+/-0.5 mV, an effect unchanged by 500 nM ouabain but abolished by 500 microM ouabain. 7. We conclude that under basal conditions, Na(+)/K(+)-ATPases containing alpha(2)- and/or alpha(3)-subunits are partially responsible for the observed K(+)-induced effects. The opening of myocyte K(+) channels (by levcromakalim or phenylephrine) creates a 'K(+) cloud' around the cells which fully activates Na(+)/K(+)-ATPase and thereby abolishes further responses to [K(+)](o) elevation.