Nutritional approaches to ameliorate pulmonary hypertension in broiler chickens.

This article reviews recent nutritional approaches for counteracting the development of pulmonary hypertension syndrome (PHS; ascites) in broiler chickens especially when they are reared at high altitudes. High altitudes impose the sustained stress of hypobaric hypoxia, which reduces the availability of atmospheric oxygen to red blood cells passing through the lungs, thereby causing systemic arterial hypoxaemia (undersaturation of haemoglobin with oxygen), pulmonary arterial hypertension and PHS/ascites in susceptible broilers. Proper nutritional strategies are needed to reduce metabolic activity and prevent the development of ascites especially when modern broilers are reared in regions where the existing altitudes limit the availability of atmospheric oxygen. This article also addresses controversies with regard to broiler nutrition in relation to PHS. For example, the catabolism of protein from feed ingredients incurs increased oxygen consumption, suggesting that feeding reduced-protein diets to broiler chickens may result in reduced PHS incidences. However, experimental and field data indicate that feeding reduced-protein diets to broilers subjected to hypobaric hypoxia increases the development of PHS. Controversies on the nutrition of unsaturated fat in relation to PHS are also discussed. In conclusion, hypoxia, acidosis, vasoconstriction and enhanced metabolic rate are triggers of PHS. Feeding reduced-protein diets might promote the susceptibility of broilers to PHS by decreased dietary intake of arginine, decreased uric acid production and increased lipogenesis. Feeding high-protein diets, dietary arginine supplementation, partial substitution of sodium bicarbonate for sodium chloride, feeding low-fat diets and effective feed restriction programmes can be considered as nutritional approaches to prevent PHS.

Efficacy of 25-OH Vitamin D3 prophylactic administration for reducing lameness in broilers grown on wire flooring.

Bacterial chondronecrosis with osteomyelitis (BCO) is the most common cause of lameness in commercial broilers. Growing broilers on wire flooring provides an excellent experimental model for reproducibly triggering significant levels of lameness attributable to BCO. In the present study we evaluated the efficacy of adding HyD (25-OH vitamin D3) to the drinking water as a preventative/prophylactic treatment for lameness. Broiler chicks were reared on 5 x 10 ft flat wire floor panels within 6 environmental chambers. Three chambers were supplied with tap water (Control group) and the remaining chambers were supplied with HyD (HyD group: 0.06 mL HyD solution/L water; dosing based on the HyD Solution label to provide 33.9 µg 25-OHD3/L) from d 1 through 56. Feed was provided ad libitum and was formulated to meet or exceed minimum standards for all ingredients, including 5,500 IU vitamin D3/kg. Lameness initially was detected on d 28, and the cumulative incidence of lameness on d 56 was higher in the Control group than in the HyD group (34.7 vs. 22.7%, respectively; P = 0.03; Z-test of proportions; chambers pooled). The most prevalent diagnoses for lame birds were osteochondrosis and osteomyelitis (BCO) of the proximal femora (52%) and tibiae (79%), accompanied by minor incidences of tibial dyschondroplasia (0.33%), spondylolisthesis, or kinky back (0.67%), and twisted legs (1%). Broilers that survived to d 56 without developing lameness did not differ in BW when compared by group within a gender. The wire flooring model imposes a rigorous, sustained challenge that undoubtedly is much more severe than typically would be experienced by broilers under normal commercial conditions. Therefore the encouraging response to HyD supplementation in the present study supports the potential for 25-OH vitamin D3 to attenuate outbreaks of lameness caused by BCO in commercial broiler flocks.

Plexogenic arteriopathy in broiler lungs: Evaluation of line, age, and sex influences.

Plexiform lesions form in the terminal pulmonary arterioles of human patients suffering from prolonged pulmonary arterial hypertension. Plexiform lesions also develop in broiler lungs, but lesion incidences are not strongly correlated with sustained pulmonary hypertension as reflected by right to total ventricular weight (RVTV) ratios. The present study was conducted to assess plexiform lesion incidences in broiler lines that have been divergently selected for susceptibility or resistance to pulmonary hypertension. Broilers from susceptible (SUS) and resistant (RES) lines were reared together and only clinically healthy (nonascitic, noncyanotic) individuals were evaluated to minimize potential line differences in cardiopulmonary hemodynamics. The objective was to determine if an innate genetic predisposition for plexogenic arteriopathy would be exposed in SUS broilers when compared with RES broilers in the absence of extreme differences in cardiopulmonary hemodynamics. Broilers up to 12 wk age from the SUS and RES lines had essentially equivalent BW, indices of cardiopulmonary function (left ventricle + septum weight, total ventricle weight, and RVTV ratios), and lung volumes within a sex. Average RVTV ratios for broilers from both lines were indicative of normal pulmonary arterial pressures at all ages sampled. Nevertheless, plexiform lesions were detected in SUS and RES broiler lungs immediately posthatch and thereafter at all ages sampled. Lesion incidences were consistently low and did not differ between the lines within any of the sampling ages. This evidence demonstrates that plexiform lesions develop extremely rapidly in broiler chicks, apparently without the prerequisite for vascular stress caused by severe, prolonged pulmonary arterial hypertension. No innate genetic predisposition for complex vascular lesion development appeared to exist in the SUS line when compared with the RES line.

Prophylactic administration of a combined prebiotic and probiotic, or therapeutic administration of enrofloxacin, to reduce the incidence of bacterial chondronecrosis with osteomyelitis in broilers.

Bacteria entering the bloodstream via translocation from the gastrointestinal tract spread hematogenously and can trigger bacterial chondronecrosis with osteomyelitis (BCO) by infecting osteochondrotic microfractures in the epiphyseal-physeal cartilage of the proximal femora and tibiae. In experiment 1, broilers were fed control feed or the same feed containing BacPack 2X, which includes the prebiotic IMW50 (a mannan oligosaccharide beta-glucan yeast cell wall product) plus the probiotic Calsporin (Bacillus subtilis C-3102). Broilers reared on wire flooring consistently developed higher incidences of BCO than hatchmates reared on wood shavings litter (≥24 vs. ≤4%, respectively; P=0.001). Adding BacPack 2X to the feed on d 1 through 56 delayed the age of onset and reduced the cumulative incidence of BCO on wire flooring when compared with broilers fed the control feed (24.0 vs. 40.7%, respectively; P=0.003). In experiment 2, broilers reared on wire flooring received tap water on d 1 through 62 (control group) or therapeutic levels of the potent fluoroquinolone antimicrobial enrofloxacin in the water on d 35 through 54 (enrofloxacin group). During enrofloxacin administration, half as many birds developed BCO in the enrofloxacin group when compared with the control group (8.1 vs. 19.5%, respectively, on d 35 through 54; P=0.001), whereas both groups had similar BCO incidences subsequent to withdrawing enrofloxacin on d 55 through 62 (14.8 vs. 18.2% for the enrofloxacin vs. control groups; P=0.386). Cumulative lameness incidences for d 1 through 62 were higher for the control group than for the enrofloxacin group (39.0 vs. 25.8%, respectively; P=0.003). These results demonstrate that wire flooring imposes a rigorous challenge that leads to high incidences of BCO that can be difficult to suppress, even with therapeutic doses of enrofloxacin. Prophylactically adding BacPack 2X to the feed reduced the incidence of BCO lameness by a proportion similar to that achieved with enrofloxacin, indicating that probiotics potentially can provide effective alternatives to antibiotics for reducing BCO lameness attributable to bacterial translocation and hematogenous distribution.

Evaluating portable wire-flooring models for inducing bacterial chondronecrosis with osteomyelitis in broilers.

Rearing broilers on flat or sloping wire flooring is an effective method for consistently triggering lameness attributable to bacterial chondronecrosis with osteomyelitis (BCO). Portable obstacles known as speed bumps (SB) also consistently trigger modest incidences of BCO when they are installed between feed and water lines in litter flooring facilities. Two experiments were conducted to determine the most effective broiler age for introducing the SB into litter flooring pens, and to evaluate alternative configurations of the traditional SB with the expectation that amplified mechanical challenges to the legs of broilers should increase the incidence of BCO. Broiler chicks obtained from commercial hatcheries (lines B and D in experiment 1, lines A and B in experiment 2) were reared in floor pens with ad libitum feed and water and a 23L:1D photoperiod. In experiment 1, the 5 floor treatments included wood shavings litter only (L), flat wire only (W), or litter plus SB installed at 14, 28, or 42 d of age. Line B was more susceptible to lameness than line D (25.9 vs. 15.3% for all treatments combined; P = 0.001). Both lines developed low incidences of lameness on L (11 to 13%), intermediate incidences on SB regardless of day of installation (12 to 23%), and high incidences on W (21 to 39%). In experiment 2, broilers were reared with 7 floor treatments, including L, W, SB with a 50% slope (SB50%); SB50% with a limbo bar installed over the apex; SB with a 66% slope and limbo bar; SB50% with a nipple water line suspended over the apex; and a pagoda-top SB. All SB were inserted on d 28. Line B was more susceptible to lameness than line A (20.2 vs. 16.1% for all treatments combined; P < 0.05), and for both lines combined the lameness percentages averaged 7.7 (L), 29.2 (W), 17.3 (SB50%), 16.2 (SB50% with a limbo bar), 21.5 (SB with a 66% slope and limbo bar), 20.8 (SB50% with a nipple water line), and 11.5% (pagoda-top). These studies demonstrate the portable SB can be effectively used to experimentally trigger BCO in broilers.

Kinetic examination of femoral bone modeling in broilers.

Lameness in broilers can be associated with progressive degeneration of the femoral head leading to femoral head necrosis and osteomyelitis. Femora from clinically healthy broilers were dissected at 7 (n = 35, 2), 14 (n = 32), 21 (n = 33), 28 (n = 34), and 42 (n = 28) d of age, and were processed for bone histomorphometry to examine bone microarchitecture and bone static and dynamic properties in the secondary spongiosa (IISP) of the proximal femoral metaphysis. Body mass increased rapidly with age, whereas the bone volume to tissue volume ratio remained relatively consistent. The bone volume to tissue volume ratio values generally reflected corresponding values for both mean trabecular thickness and mean trabecular number. Bone metabolism was highest on d 7 when significant osteoblast activity was reflected by increased osteoid surface to bone surface and mineralizing surface per bone surface ratios. However, significant declines in osteoblast activity and bone formative processes occurred during the second week of development, such that newly formed but unmineralized bone tissue (osteoid) and the percentages of mineralizing surfaces both were diminished. Osteoclast activity was elevated to the extent that measurement was impossible. Intense osteoclast activity presumably reflects marked bone resorption throughout the experiment. The overall mature trabecular bone volume remained relatively low, which may arise from extensive persistence of chondrocyte columns in the metaphysis, large areas in the metaphysis composed of immature bone, destruction of bone tissue in the primary spongiosa, and potentially reduced bone blood vessel penetration that normally would be necessary for robust development. Delayed bone development in the IISP was attributable to an uncoupling of osteoblast and osteoclast activity, whereby bone resorption (osteoclast activity) outpaced bone formation (osteoblast activity). Insufficient maturation and mineralization of the IISP may contribute to subsequent pathology of the femoral head in fast-growing broilers.

Bacterial chondronecrosis with osteomyelitis in broilers: influence of sires and straight-run versus sex-separate rearing.

Two experiments (E1, E2) were conducted to compare the influence of sires (sire A on dam C vs. sire B on dam C) and straight-run versus sex-separate rearing on the incidence of bacterial chondronecrosis with osteomyelitis (BCO) in broilers. Fertile eggs from commercial breeder flocks were incubated and hatched at the University of Arkansas Poultry Research Hatchery. Male and female chicks were reared together (straight-run) or separately (sex-separate) in 3 × 3 m pens on litter or flat wire flooring with 65 (E1) or 60 (E2) birds per pen. Necropsies revealed lesions that are pathognomonic for BCO in ≥98% of the birds that became lame. The SigmaStat Z-test was used to compare cumulative BCO incidences through 8 wk of age. For birds reared on litter, the incidences of BCO were low regardless of cross or sex (range: 1.7 to 5.1%; P ≥ 0.6). Within a cross and sex, rearing the broilers straight-run versus sex-separate on wire flooring did not significantly affect the incidence of BCO. Significant incidences of BCO did not develop until after d 40. Males from the sire A cross developed a higher incidence of BCO than males from the sire B cross in E1 (27 vs. 17%, respectively; P = 0.009) but not in E2 (28.5 vs. 22.6%, respectively; P = 0.141). In both experiments, males from the sire A cross developed higher incidences of BCO than females from the sire B cross (27 vs. 11.9%, in E1; 28.5 vs. 14.8%, in E2). With the sexes pooled, broilers from the sire A cross consistently developed higher incidences of BCO than broilers from the sire B cross (21.4 vs. 14.9%, P = 0.005 in E1; 26.5 vs. 18.7%, P = 0.003 in E2). High susceptibilities to both femoral head (all femoral head necrosis = 66 to 85% incidences) and tibial head (all tibial head necrosis = 81 to 96% incidences) BCO lesions were demonstrated in lame birds from both sexes and crosses. This study supports a sire influence on the susceptibility of broilers to BCO. Sire lines can be chosen to reduce BCO susceptibility when broilers are grown beyond 6 wk of age.

Pulmonary arterial hypertension (ascites syndrome) in broilers: a review.

Pulmonary arterial hypertension (PAH) syndrome in broilers (also known as ascites syndrome and pulmonary hypertension syndrome) can be attributed to imbalances between cardiac output and the anatomical capacity of the pulmonary vasculature to accommodate ever-increasing rates of blood flow, as well as to an inappropriately elevated tone (degree of constriction) maintained by the pulmonary arterioles. Comparisons of PAH-susceptible and PAH-resistant broilers do not consistently reveal differences in cardiac output, but PAH-susceptible broilers consistently have higher pulmonary arterial pressures and pulmonary vascular resistances compared with PAH-resistant broilers. Efforts clarify the causes of excessive pulmonary vascular resistance have focused on evaluating the roles of chemical mediators of vasoconstriction and vasodilation, as well as on pathological (structural) changes occurring within the pulmonary arterioles (e.g., vascular remodeling and pathology) during the pathogenesis of PAH. The objectives of this review are to (1) summarize the pathophysiological progression initiated by the onset of pulmonary hypertension and culminating in terminal ascites; (2) review recent information regarding the factors contributing to excessively elevated resistance to blood flow through the lungs; (3) assess the role of the immune system during the pathogenesis of PAH; and (4) present new insights into the genetic basis of PAH. The cumulative evidence attributes the elevated pulmonary vascular resistance in PAH-susceptible broilers to an anatomically inadequate pulmonary vascular capacity, to excessive vascular tone reflecting the dominance of pulmonary vasoconstrictors over vasodilators, and to vascular pathology elicited by excessive hemodynamic stress. Emerging evidence also demonstrates that the pathogenesis of PAH includes characteristics of an inflammatory/autoimmune disease involving multifactorial genetic, environmental, and immune system components. Pulmonary arterial hypertension susceptibility appears to be multigenic and may be manifested in aberrant stress sensitivity, function, and regulation of pulmonary vascular tissue components, as well as aberrant activities of innate and adaptive immune system components. Major genetic influences and high heritabilities for PAH susceptibility have been demonstrated by numerous investigators. Selection pressures rigorously focused to challenge the pulmonary vascular capacity readily expose the genetic basis for spontaneous PAH in broilers. Chromosomal mapping continues to identify regions associated with ascites susceptibility, and candidate genes have been identified. Ongoing immunological and genomic investigations are likely to continue generating important new knowledge regarding the fundamental biological bases for the PAH/ascites syndrome.

Susceptibility of 4 commercial broiler crosses to lameness attributable to bacterial chondronecrosis with osteomyelitis.

Growing broilers on wire flooring provides an excellent experimental model for exposing susceptibility to lameness attributable to bacterial chondronecrosis with osteomyelitis (BCO). Two independent experiments (E1, E2) were designed to compare the susceptibilities of broilers from 4 commercial crosses (W, X, Y, and Z). The standard crosses (W and Y) grow rapidly at an early age, whereas high-yield crosses (X and Z) initially tend to grow more slowly. Chicks were obtained from a commercial hatchery for E1, or were hatched at the University of Arkansas Poultry Research Hatchery for E2. Males and females were reared together (E1; n = 360/cross) or separately (E2; n = 390/cross) in 3 × 3 m pens on litter or wire flooring (wire). Necropsies revealed lesions that were pathognomonic for BCO in ≥94% of the birds that became lame. The SigmaStat Z-test was used to compare cumulative lameness incidences at 8 wk of age. For birds reared on litter, lameness incidences were low and did not differ between crosses or sexes (range: 2.2 to 4.6%; P ≥ 0.6). When males were reared on wire, their lameness incidences (by cross) were E1 = 52% for W(b); 42% for X(c); 69% for Y(a), and 44% for Z(bc); E2 = 31% for W(b); 19% for X(c); 49% for Y(a); and 25% for Z(bc). For females reared on wire, the lameness incidences were E1 = 40% for W(b), 30% for X(c), 49% for Y(a), and 28% for Z(c); E2 = 16% for W; 15% for X; 16% for Y; and 15% for Z (ns). Accordingly, the hierarchical ranking for BCO susceptibility by broiler cross was X ≤ Z ≤ W < Y for males in E1 and E2, for females in E1, and for males and females pooled in E1 and E2. Standard broiler crosses developed higher incidences of lameness than high-yield crosses, implicating an association between rapid early growth and susceptibility to BCO. Rearing the females separately on wire in E2 led to uniformly low incidences of BCO, regardless of cross. Stress-mediated immunosuppression contributes to the pathogenesis of BCO; perhaps female broilers experience less social or competitive stress when reared separately from their male hatch mates.

Dexamethasone triggers lameness associated with necrosis of the proximal tibial head and proximal femoral head in broilers.

Bacterial chondronecrosis with osteomyelitis (BCO) and turkey osteomyelitis complex (TOC) are characterized by bacterial infection and necrotic degeneration within the tibiae and femora. Stress and immunosuppression have been implicated in the pathogenesis of BCO and TOC. Immunosuppressive doses of dexamethasone (DEX) trigger high incidences of TOC in turkey poults. The present study was conducted to determine if DEX injections or heat stress can trigger BCO and lameness in broilers. In 3 independent experiments, broilers were weighed and either remained uninjected or received repeated injections of 0.9% saline or DEX dissolved in saline (0.45 to 1.5 mg of DEX/kg of BW). Across all 3 experiments, the incidences of lameness were 0% for uninjected controls, 0 to 8% in saline-injected groups, and 24 to 68% in groups injected with 0.9 to 1.5 mg of DEX/kg of BW. Growth was inhibited by DEX injections regardless of whether the birds became lame or survived. When compared with saline-injected groups, DEX injections consistently increased the incidence of severe proximal tibial head necrosis in lame birds as well as in survivors. The DEX injections also triggered a subset of lesions that are not considered pathognomonic for BCO (for example, avascular femoral head necrosis and fatty necrosis of the tibiae). In a fourth experiment, repeated episodes of heat stress did not trigger lameness, although the subclinical incidence of tibial head necrosis was substantially higher at 28 and 35 d of age in heat-stressed broilers when compared with broilers reared under thermoneutral conditions. Accordingly, stress and immunosuppression must be considered contributing factors in the pathogenesis of tibial and femoral lesions associated with lameness in broilers. A subset of the lesions triggered by repeated DEX injections did not precisely mimic the pathogenesis of BCO in broilers, and DEX consistently inhibited growth whereas BCO is associated with rapid growth. These caveats must be acknowledged when DEX is used to trigger lameness in broilers.

Intrapulmonary arteries respond to serotonin and adenosine triphosphate in broiler chickens susceptible to idiopathic pulmonary arterial hypertension.

This study examined factors contributing to increased vascular resistance and plexiform lesion formation in broiler chickens susceptible to idiopathic pulmonary arterial hypertension (IPAH). A diet supplemented with excess tryptophan (high-Trp diet), the precursor for serotonin, was used to accelerate the development of IPAH. Broilers fed the high-Trp diet had higher pulmonary arterial pressures than broilers fed the control diet, and plexiform lesion incidences tended to be higher (P = 0.11) in the high-Trp group than in the control group at 30 d of age. The intrapulmonary arteries were assessed for vasoconstriction in response to serotonin and adenosine triphosphate (ATP) and for activities of key metabolic enzymes for serotonin and ATP. The pulmonary artery (defined as the first major branch of the pulmonary artery inside the lung) and the primary pulmonary arterial rami (defined as the second major branch of the pulmonary artery inside the lung) both exhibited vasoconstriction in response to serotonin and ATP. This is the first study to demonstrate purinergic-mediated vasoconstriction in intrapulmonary arteries from broilers. Arteriole responsiveness did not differ between broilers fed the control diet or the high-Trp diet. Therefore, the high-Trp diet enhanced the development of IPAH but did not affect the artery's sensitivity to serotonin or ATP. Monoamine oxidase activity, responsible for the breakdown of serotonin, was severely impaired in pulmonary arteries from broilers in the high-Trp group. Accordingly, serotonin may persist longer and elicit an amplified response in broilers fed the high-Trp diet.

A wire-flooring model for inducing lameness in broilers: evaluation of probiotics as a prophylactic treatment.

Bacterial chondronecrosis with osteomyelitis (BCO) is the most common cause of lameness in commercial broilers. Bacteria entering the blood via translocation from the respiratory system or gastrointestinal tract spread hematogenously to the proximal epiphyseal-physeal cartilage of rapidly growing femora and tibiae, causing BCO. We tested the hypothesis that rearing broilers on wire flooring should increase the incidence of BCO by persistently imposing additional torque and shear stress on susceptible leg joints. We also tested the hypothesis that probiotics might attenuate bacterial translocation and thereby reduce the incidence of BCO. In 5 independent experiments using 4 commercial lines, broilers grown on wire flooring developed lameness attributable predominately to BCO. The fastest-growing birds were not necessarily the most susceptible to lameness on wire flooring, nor did the genders differ in susceptibility in the 2 experiments that included both male and female broilers. The pathogenesis of BCO is not instantaneous, and accordingly, many broilers that did not exhibit lameness, nevertheless, did possess early pathognomonic lesions. These subclinical lesions were equally likely to develop in the right or left leg. The lesion status of the proximal femoral head did not determine the lesion status of the ipsilateral or contralateral proximal tibial head and vice versa. Broilers reared on wire flooring consistently had higher incidences of lameness than hatch-mates reared on wood-shavings litter. Adding probiotics to the diet beginning at 1 d of age consistently reduced the incidence of lameness for broilers reared on wire flooring. These experiments indicate that probiotics administered prophylactically may constitute an alternative to antibiotics for reducing lameness attributable to BCO. Rearing broilers on wire flooring provides an important new research model for investigating the etiology, pathogenesis, and treatment strategies for BCO.

Methylglyoxal and pulmonary hypertension in broiler chickens.

Methylglyoxal (MG) is a dicarbonyl molecule that forms during glycolysis and normally is detoxified via the glyoxalase system. Methylglyoxal is highly reactive with various amino acid residues in proteins, leading to oxidative stress and irreversible protein damage. Increased levels of MG have been associated with endothelial damage and vascular remodeling contributing to the development of systemic arterial hypertension in mammals. This study was conducted to determine whether administering exogenous MG can trigger pulmonary hypertension (increased pulmonary arterial pressure) in broilers. Hematological assays and preliminary mass spectrometric analyses also were conducted using blood samples from broilers that had been injected intramuscularly with either saline or MG to determine whether MG triggers either a toxic response or oxidative posttranslational modification of hemoglobin within 24 h postinjection. Clinically healthy male broilers received 100-µL intravenous injections of saline and then MG, followed by a 500-µL intramuscular injection. Neither intravenous nor intramuscular injections of saline altered the pulmonary arterial pressure, whereas both intravenous and intramuscular MG injections triggered pulmonary hypertension attributable to increased pulmonary vascular resistance. The precise mode of action by which MG triggers pulmonary vasoconstriction remains to be determined. Pulse oximetry, hematology, and matrix-assisted laser desorption ionization-time-of-flight spectra data did not provide evidence of an overt toxic response to MG, nor was modification of hemoglobin detected, although increased heterophil:lymphocyte ratios did demonstrate that MG caused a stress response. To the best of our knowledge the present results constitute the first demonstration in any vertebrate species that exogenously administered MG rapidly initiates pulmonary hypertension attributable to pulmonary vasoconstriction.

Effects of supplementation of canola meal-based diets with arginine on performance, plasma nitric oxide, and carcass characteristics of broiler chickens grown at high altitude.

A total of 300 male broilers (Ross 308) were exposed to cool conditions at high altitudes to study the effects of dietary Arg supplementation on performance and physiological and zootechnical variables. A corn-soybean meal (SBM) and a corn-canola meal (CM) diet were formulated for the starting (1 to 3 wk of age) and growing (3 to 6 wk of age) stages according to NRC recommendations. Two additional diets were prepared by supplementing 0.2 and 0.4% l-Arg to the corn-CM diet. Substitution of CM for SBM caused a significant (P < 0.05) reduction in weight gain and feed intake and resulted in impaired feed:gain. Supplementing Arg in the CM diet restored the feed and weight losses to a significant extent so that a significant difference was found between CM diet and CM + 0.4% Arg in terms of weight gain for the growing (3 to 6 wk) stage and the entire study (1 to 6 wk; P < 0.05). Total plasma nitric oxide (NO) concentration analyzed by nitrate plus nitrite assay was measured in the treatment groups. A significant (P < 0.05) decrease in plasma NO level was observed by substituting CM for SBM in the diet. Supplementing the CM diet with Arg increased the plasma NO level above that of SBM group. Carcass and breast yields were significantly decreased (P < 0.05) as a result of substituting CM for SBM. The substitution of CM for SBM, however, significantly (P < 0.05) increased the proportions of thighs and heart. The right ventricular weight:total ventricular weight ratio and ascites mortality showed a significant (P < 0.05) increase when SBM was replaced by CM in the diet. Fortification of the CM diet with Arg eliminated the significant difference in the right-to-total ventricular weight ratios when compared with the SBM diet. In conclusion, feeding CM to broiler chickens raised at high altitude caused reduced growth performance and predisposed the birds to pulmonary hypertension and ascites, which were partly restored by Arg supplementation.

Pulmonary vascular pressure profiles in broilers selected for susceptibility to pulmonary hypertension syndrome: age and sex comparisons.

Broilers that are susceptible to pulmonary hypertension syndrome (PHS, ascites) have an elevated pulmonary arterial pressure (PAP) when compared with PHS-resistant broilers. Two distinctly different syndromes, pulmonary arterial hypertension and pulmonary venous hypertension (PVH), both are associated with increases in PAP. Pulmonary arterial hypertension occurs when the right ventricle must elevate the PAP to overcome increased resistance to flow through restrictive pulmonary arterioles upstream from the pulmonary capillaries. In contrast, PVH is commonly caused by increased downstream (postcapillary) resistance. The sites of resistance to pulmonary blood flow are deduced by making contemporaneous measurements of the PAP and the wedge pressure (WP) and calculating the transpulmonary pressure gradient (TPG) (TPG = PAP - WP). We obtained PAP and WP values from 8-, 12-, 16-, 20-, and 24-wk-old anesthetized male and female broilers from a PHS-susceptible line. Pressures were recorded as a catheter was advanced through a wing vein to the pulmonary artery and onward until the WP was obtained. In addition to sex and age comparisons of vascular pressure gradients, the data also were pooled to obtain 3 cohorts for broilers having the lowest PAP values (n = 52; range: 12 to 22.9 mmHg), intermediate PAP values (n = 63; range: 23 to 32.9 mmHg), and highest PAP values (n = 62; range: 33 to 62 mmHg) independent of age or sex. Within each of the age, sex, and PAP cohort comparisons, broilers with elevated PAP consistently exhibited the hemodynamic characteristics of pulmonary arterial hypertension (elevated PAP and TPG combined with a normal WP) and not PVH (elevated PAP and WP combined with a normal or reduced TPG). Susceptibility to PHS can be attributed primarily to pulmonary arterial hypertension associated with increased precapillary (arteriole) resistance.

Broiler pulmonary hypertensive responses during lipopolysaccharide-induced tolerance and cyclooxygenase inhibition.

Bacterial lipopolysaccharide (LPS, endotoxin) triggers pulmonary hypertension (PH) characterized by an increase in pulmonary arterial pressure (PAP) that reaches a peak value within 20 to 25 min and then gradually subsides within 60 min. As the PAP subsides PH cannot be reinitiated, signifying the onset of a period of tolerance (refractoriness) to repeated LPS exposure. The present study was conducted to determine the duration of this tolerance, and to evaluate key mediators thought to contribute to LPS-mediated PH in broilers. Tolerance was shown to persist for 4 to 5 d after the initial exposure to LPS. In tolerant broilers supramaximal i.v. injections of LPS did not reinitiate PH, nor was a significant modulatory role for nitric oxide demonstrated. The pulmonary vasculature of tolerant broilers remains responsive to the thromboxane A(2) (TxA(2)) mimetic U44069, 5-hydroxytryptamine (5-HT, serotonin), and constitutive nitric oxide. Meclofenamate successfully blocked the conversion of arachidonic acid to vasoconstrictive eicosanoids such as TxA(2); nevertheless, meclofenamate failed to inhibit PH in response to LPS. Therefore, TxA(2) does not appear to be the primary vasoconstrictor involved in the PH response to LPS and neither does 5-HT. Broilers emerging from tolerance 5 d after the initial exposure to LPS exhibited interindividual variation in their PH responsiveness to a second LPS injection, ranging from zero response (individuals that remain fully tolerant) to large increases in PAP (post-tolerant individuals). Tolerance might be an important compensatory or protective mechanism for broilers whose pulmonary vascular capacity is marginally adequate under optimal conditions, and whose respiratory systems are chronically challenged with LPS in commercial production facilities. The key vasoconstrictors responsible for the PH elicited by LPS remain to be determined.

Cellular component of lavage fluid from broilers with normal versus aerosol-primed airways.

Previously, we reported that intratracheal administration of lipopolysaccharide elicited pulmonary hypertension (PH) in broilers reared under commercial conditions and in broilers reared in environmental chambers and pretreated with aerosolized red food colorant # 3 and propylene glycol (Red#3+PG), but not in control broilers reared in environmental chambers. The objective of the present experiment was to determine possible changes in the number or proportion of airway leukocytes that could contribute to the magnitude of the PH responses elicited in broilers. Birds were aerosolized for 40 min with a saturated mixture of Red#3+PG. After 24 h, a blood sample was taken, the broilers were killed, and a pulmonary lavage process was conducted in each bird. Leukocyte concentration (white blood cells/microL) and differential leukocyte counts (%) were measured in blood and lavage fluid. Leukocyte concentration in blood did not differ between groups, but the percentage of blood lymphocytes was lower in broilers from the Red#3+PG group compared with birds from the control group (52.4+/-2.9 and 56.9+/-2.9%, respectively). Cells recovered from the lavage fluid from both groups were primarily heterophils. The concentration of leukocytes was greater in the lavage fluid of broilers from the Red#3+PG group compared with broilers from the control group (763.2+/-158.7 and 402.9+/-62.6 white blood cells/microL, respectively), but the proportions among leukocytes were not different between the 2 groups. We propose that the increased concentration of leukocytes present within the airways was one of the components that enabled broilers pre-treated with aerosolized Red#3+PG to exhibit PH responses to intratracheal lipopolysaccharide.

Pulmonary hemodynamic responses to intravenous prostaglandin E2 in broiler chickens.

Prostaglandin E(2) (PGE(2)) affects pulmonary arterial pressure (PAP), pulmonary vascular resistance (PVR), and respiratory rate (RR) in mammals, but no information previously was available regarding avian pulmonary responses to PGE(2). Two experiments were conducted in which 45- to 55-d-old male broiler chickens were infused i.v. with PGE(2) at the lowest rate (30 mug/min for 4 min) that reliably reduced PAP during pilot studies. When compared with preinfusion (control) values in experiment 1, PGE(2) reduced PAP from 19 +/- 1 to 16 +/- 1 mmHg (P < 0.001) and reduced mean systemic arterial pressure from 111 +/- 6 to 81 +/- 5 mmHg (P < 0.001) but did not significantly reduce heart rate (HR; control: 338 +/- 9 beats/min; PGE(2): 320 +/- 12 beats/min; P > 0.05). Infusing PGE(2) also reduced the RR from 57 +/- 2 to 46 +/- 4 breaths/min (P < 0.001) and reduced the percentage saturation of hemoglobin with oxygen (%HbO(2)) from 85 +/- 2 to 77 +/- 3%HbO(2) (P < 0.001). After the PGE(2) infusion ceased, the PAP, mean systemic arterial pressure, RR, and %HbO(2) recovered within 8 min to levels that did not differ from preinfusion control values. In experiment 2, an ultrasonic flow probe was surgically implanted on 1 pulmonary artery to measure cardiac output (CO). When compared with preinfusion control values, PGE(2) reduced CO from 140 +/- 6 to 111 +/- 5 mL/kg of BW x min (P < 0.001), reduced PAP from 25 +/- 2 to 21 +/- 1 mmHg (P < 0.001), and reduced RR from 49 +/- 4 to 35 +/- 4 breaths/min (P < 0.001). The reduction in CO was caused by a reduction in HR from 305 +/- 9 to 260 +/- 9 beats/min without a significant reduction in stroke volume (control: 0.46 +/- 0.02 mL/kg of BW x beat; PGE(2): 0.43 +/- 0.02 mL/kg of BW x beat; P = 0.158). After the PGE(2) infusion ceased the CO, PAP, RR, and HR recovered within 9 min to levels that did not differ from preinfusion control values. The PVR, calculated as PAP/CO, was not altered by PGE(2) (control: 0.18 +/- 0.01 relative resistance units; PGE(2): 0.20 +/- 0.02 relative resistance units; P > 0.723). These results indicate that in broilers PGE(2) reduced PAP by reducing CO rather than by acting as a pulmonary vasodilator to lower PVR. The PGE(2)-induced reductions in PAP would benefit broilers that are susceptible to pulmonary hypertension syndrome by reducing their right ventricular overload; however, the reductions in CO and RR combined with the onset of systemic arterial hypoxemia would accelerate the pathophysiological progression leading to terminal pulmonary hypertension syndrome.

Inhaling one hundred percent oxygen eliminates the systemic arterial hypoxemic response of broilers to intravenous microparticle injections.

The pathogenesis of pulmonary hypertension syndrome (PHS, ascites) includes the development of systemic arterial hypoxemia (reduction in the saturation of hemoglobin with O(2), HbO(2)), which can be mimicked in clinically healthy broilers by i.v. injections of microparticles (MP). In experiment 1, arterial blood samples were collected from clinically healthy broilers before and after i.v. MP injections, and during a subsequent 100% O(2) inhalation period. The arterial samples were analyzed for HbO(2), partial pressure of O(2) and CO(2), and pH using a blood gas analyzer. In experiment 2, broilers that initially averaged > or =75% HbO(2) were assigned to a "high O(2)" group, whereas those that initially averaged < 75% HbO(2) were assigned to a "low O(2)" group. The HbO(2) and heart rate (HR) were measured using a pulse oximeter before, during, and after broilers in both groups inhaled 100% O(2). In experiment 3, HbO(2) and HR were measured using a pulse oximeter before, during, and after broilers inhaled 100% O(2), after i.v. MP injections, and during a second period of 100% O(2) inhalation. The HbO(2) rapidly decreased after i.v. MP injections, and subsequently providing 100% O(2) to breathe increased the HbO(2) above preinjection control levels in experiments 1 and 3. In experiment 2, inhaling 100% oxygen eliminated the initial spontaneous differences in HbO(2) between the high O(2) and low O(2) groups, whereas the return to breathing ambient air restored the initial group differences in HbO(2). These experiments indicate that MP-induced and spontaneous hypoxemia can be attributed to a diffusion limitation rather than to arterial-venous shunts, because the hypoxemia resulting from arterial-venous shunts cannot be wholly eliminated by providing 100% O(2) to inhale.

Intratracheal administration of bacterial lipopolysaccharide elicits pulmonary hypertension in broilers with primed airways.

Broilers reared under commercial conditions inhale irritant gases and aerosolized particulates contaminated with gram-negative bacteria and bacterial lipopolysaccharide (LPS). Previous studies demonstrated that i.v. injections of LPS can trigger an increase in the pulmonary arterial pressure (PAP); however, the pulmonary hemodynamic response to aerosolized LPS entering via the most common route, the respiratory tract, had not been evaluated in broilers. In experiment 1, broilers reared on new wood shavings litter in clean environmental chambers either were not pretreated (control group) or were pretreated via aerosol inhalation of substances (food color dyes and propylene glycol) known to sensitize the airways. One day later, the broilers were anesthetized, catheterized to record the PAP, and an intratracheal aerosol spray of LPS (1 mL of 2 mg/mL of LPS) was administered. Broilers in the control group as well as broilers pretreated with aerosolized distilled water or yellow and blue food color dyes did not develop pulmonary hypertension (PH; an increase in PAP) after the intratracheal spray of LPS, whereas broilers that had been pretreated with red food color did develop PH in response to intratracheal LPS. In experiment 2, birds raised under commercial conditions on used wood shavings litter developed PH in response to intratracheal LPS regardless of whether they had been pretreated with aerosolized red food color dye. In experiment 3, broilers reared in clean environmental chambers on new wood shavings litter were used to demonstrate that Red Dye #3 and propylene glycol are capable of priming the responsiveness of the airways to a subsequent intratracheal LPS challenge. Common air contaminants such as LPS can result in PH leading to pulmonary hypertension syndrome (ascites) in broilers with appropriately primed airways.