Effects of treatments with cyclooxygenase inhibitors on chickens infected with Eimeria acervulina.

Reduced weight gains and feed conversions are major sources of economic losses from avian coccidiosis. Experiments were conducted to determine possible involvement of up-regulation of cyclooxygenase (COX) activity during coccidia infections that might contribute to reduced weight gain. In a series of trials, young (3 to 5 wk of age) cockerels infected with Eimeria acervulina, a duodenal parasite, received oral treatments with indomethacin (IM), an inhibitor of both COX-1 and COX-2, or nimesulide (NIM), a specific inhibitor of COX-2. Infection doses varied with experiment (from 10(5) to 10(6) oocysts per chick). Effects of infection on weight gain, duodenal lesions, plasma carotenoids, and levels of NO2+ NO3- were determined at 6 d postinoculation. Total oocysts were enumerated in feces collected from 5 through 8 d postinoculation from separate replicate groups. In no trials did treatment with IM reverse effects of infection on weight gain or significantly reduce lesion scores. However, in all trials, IM treatment reduced oocyst output per chick (Trial 1, 15%; Trial 2, 19%; Trial 3, 53%; Trial 4, 29%; Trial 5, 28.5%). Supplementation of feed with 400 and 100 ppm NIM significantly reduced weight gain of, and increased oocyst output from, infected chicks, whereas supplementation with 50 ppm NIM had no beneficial effects on weight gain or lesion scores, but reduced oocyst shedding. The inability of the IM and NIM treatments to reverse infection-associated weight gain suppression suggests that this pathological effect is not linked to increased prostanoid synthesis as a result of COX-2 up-regulation during infection. The inhibitory effects of IM treatment on oocyst shedding suggest that COX-2 products may have immunosuppressive effects in coccidia infection at local sites of infection. The stimulating effects of high-dose NIM treatments on oocyst shedding suggest that this compound may inhibit synthesis of other prostanoids as well.

Effects of nutritionally balanced and stabilized flaxmeal-based diets on Eimeria tenella infections in chickens.

Twenty Sex Sal cockerels were randomly assigned to each of eight groups; each of four nutritionally balanced diets were fed to two groups from 1 d through 4 wk of age. These diets contained 0, 2, 5, or 10% stabilized flaxseed meal that provided a calculated 0, 0.45, 1.11, or 2.22% n-3 fatty acids, primarily linolenic acid. At 3 wk of age, one group of chickens from each diet treatment was infected with Eimeria tenella and was housed in separate but similar conditions to uninfected control chickens. At 6 d postinfection, chickens were weighed, bled, killed, and scored for lesions. No level of dietary flaxmeal tested provided protection against weight gain depression, increased feed conversion ratios, or lesions. We concluded that these diets did not protect against E. tenella infection because levels of linolenic acid were not high enough, and the oxidative potentials were well suppressed by vitamin E and other stabilizers present.

Effects of dietary supplementation with n-3 fatty acid ethyl esters on coccidiosis in chickens.

The ethyl esters of eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids were added to a broiler starter diet singly or in combination [as bulk purified ethyl ester concentrate from menhaden oil (n3FAC)] in quantities similar to those found in a diet supplemented with 5% menhaden oil (MO). Diets were fed to chickens from 1 d of age through 3 wk of age. At 2 wk of age, the chickens were infected with Eimeria tenella, Eimeria acervulina, or Eimeria maxima. At 6 d postinfection (PI), the effects of the diets were assessed on weight gains, plasma carotenoids, gross lesion scores, and histological parasite scores in gut cross sections, or oocyst output. Significant ameliorating effects of diet on lesion scores and parasite scores were only seen in E. tenella infections and were only produced by the n3FAC and MO supplements. These two supplements, which contained higher molar concentrations of double bonds than the other supplements, also significantly reduced plasma carotenoids in uninfected chickens, indicating that they promoted a state of oxidative stress. These results are consistent with previous reports on the interaction of coccidiosis with dietary n-3 fatty acids and strengthen the hypotheses that dietary-induced oxidative stress is an effective deterrent against cecal coccidiosis in chickens.

Dietary modulation of avian coccidiosis.

During the past several years, our laboratory has been investigating the anticoccidial activities of various natural products that have potential use as dietary supplements for coccidiosis control. Sources of fats containing high concentrations of n-3 fatty acids such as menhaden oil and flaxseed oil and flaxseed, when added to starter rations and fed to chicks from one day of age, effectively reduce lesions caused by the caecal parasite Eimeria tenella, but not lesions caused by Eimeria maxima. Our results are consistent with reports of effects of diets high in n-3 fatty acids on other protozoan parasites which suggest that the state of oxidative stress induced by these diets in the cells of both host and parasites is responsible for their parasitic actions. Artemisinin, a naturally occurring (Artemisia annua) endoperoxide and effective antimalarial significantly lowers lesions from E. tenella when given at low levels as a feed additive. The mechanism of its action is also considered to involve induction of oxidative stress. Diets supplemented with 8 p.p.m. gamma-tocopherol (abundant in flaxseeds) or with 1% of the spice tumeric, reduce mid-small intestinal lesion scores and improve weight gains during E. maxima infections. These compounds may exert their anticoccidial activity because they are effective antioxidants. Betaine, a choline analogue found in high concentrations in sugar beets, improves nutrient utilisation by animals under stress. When provided as a dietary supplement at a level of 0.15% it has enhanced the anticoccidial activity of the ionophore, salinomycin. Betaine may act as an osmoprotectant whereby it improves the integrity and function of the infected intestinal mucosa. In in vivo studies, betaine plus salinomycin significantly inhibit invasion of both E. tenella and E. acervulina. However, subsequent development of E. acervulina is inhibited more effectively with this combination treatment than development of E. tenella.

Effects of components of Artemisia annua on coccidia infections in chickens.

Four experiments were run to test the anticoccidial activity of dried Artemisia annua leaves and several of their chemical constituents for possible use as prophylactic feed additives. When fed over a period of 3 wk at a level of 5%, a dried leaf supplement of A. annua provided significant protection against lesions due to Eimeria tenella but not Eimeria acervulina or Eimeria maxima. When fed over a period of 5 wk at a level of 1% to chicks undergoing immunization with a live vaccine, it provided significant protection in partially immunized chicks against E. acervulina and E. tenella lesions from a dual species challenge infection. It also afforded lower mean lesion scores in challenged chicks immunized over a period of 5 wk. Artemisinin, an antimalarial component of A. annua, was present at a level of 0.034% in the dried leaf preparation. A 5% supplement thus afforded about 17 ppm artemisin. When the pure compound was fed at that level for a period of 3 wk, it protected weight gains and significantly reduced lesion scores attributable to E. tenella but not E. acervulina. Other components of A. annua, camphor and 1,8-cineole, at 119 ppm also protected weight gains, and reduced E. tenella lesion scores. Camphor reduced E. acervulina lesions. Artemisinin fed for 4 wk at levels of 2, 8.5, and 17 ppm significantly reduced oocyst output from separate E. acervulina and E. tenella infections and a dual species infection. Pure artemisinin thus appears to be effective against at least two coccidia species when used as a feed additive, and its activity may depend, in part, on the length of time it is administered before a challenge infection.

Interaction of dietary flaxseed with coccidia infections in chickens.

Two experiments were conducted to determine effects of diets containing n-3 fatty acids (n-3FA) from whole as well as ground flaxseed on the performance of broilers during coccidia infections. Diets were fed from 1 d of age through 3 wk of age. Chickens were infected with coccidia at 2 wk of age and the effects of infection assessed at 6 d postinfection. The first experiment contrasted effects of several high n-3FA-containing diets, including one supplemented with whole flaxseed, on infections with Eimeria tenella or Eimeria maxima. Infected chickens that consumed the flaxseed-supplemented diet had the lowest weight gains, but they were not significantly different from gains of infected chickens on the control diet. Diets supplemented with 5% menhaden oil or 15% flaxseed significantly reduced lesions caused by E. tenella, but had no effect on lesions caused by E. maxima. In a second experiment, diets supplemented with 5 or 10% ground flaxseed were assessed for effects on the performance of broilers infected with three dose levels (500, 5,000, or 50,000 oocysts) of E. maxima. Neither flaxseed diet protected weight gain during infection with 50,000 oocysts. However, a 5% flax diet protected weight gains in chickens infected with 500 or 5,000 oocysts. Diets supplemented with 5 or 10% ground flaxseed exacerbated lesions in chickens infected with 5,000 or 50,000 E. maxima oocysts compared to the control diet. Thus, diets containing high amounts of n-3 fatty acids do not affect the development of all Eimeria sp. in the same manner. The oxidative stress produced by these diets may more adversely affect development of E. tenella, which infects the relatively oxygen-poor ceca, whereas it does not affect development of E. maxima which parasitizes the middle portion of the small intestine.

The effect of high n-3 fatty acids diets on the ultrastructural development of Eimeria tenella.

A study of development of Eimeria tenella in chickens fed high n-3 fatty acids (n-3FA) diets showed ultrastructural degeneration of both asexual and sexual parasite stages. Abnormal shedding of asexual and sexual parasite developmental stages into the cecal lumen was also observed. Ultrastructural degeneration was characterized by cytoplasmic vacuolization, chromatin condensation within the nucleus, a lack of parasitophorous vacuole delineation, and, in some cases, a complete loss of parasite ultrastructural organization. The results of this study indicate that diets high in n-3FA may be useful in the control of avian coccidia.

Association of lowered plasma carotenoids with protection against cecal coccidiosis by diets high in n-3 fatty acids.

A series of four experiments was run to assess the effectiveness of diets containing high amounts of n-3 fatty acids in reducing the pathological effects of cecal coccidiosis in chickens caused by Eimeria tenella. To determine whether the dietary effects were related to development of oxidative stress, plasma samples were analyzed for tocopherols and carotenoids. Plasma vitamin E (alpha-tocopherol) values were not consistent between experiments. Total plasma carotenoids, however, were significantly decreased by 2.5 to 20% diet supplementation with fish oil in several experiments. These decreases coincided with significant reductions in lesion scores. Under the experimental conditions, total plasma carotenoid concentrations may serve as a sensitive indicator for oxidative stress, which may be a factor in reducing cecal lesions in E. tenella infections.

Diets high in n-3 fatty acids reduce cecal lesion scores in chickens infected with Eimeria tenella.

Four experiments were run in which diets incorporating various levels of n-3 fatty acids (n-3FA) from fish (menhaden) oil or flax seed were tested for their abilities to reduce the severity of Eimeria tenella infections in young broiler chicks. The diets were fed from 1 d of age through 3 wk of age. Chickens were infected at 2 wk of age. Diets consisting of broiler starter ration supplemented with 2.5 to 10% fish oil, 10% flax seed oil, or 10% linseed oil significantly decreased cecal lesions and maintained weight gains as compared to unsupplemented diets. Light micrographs of ceca from chickens that consumed high n-3FA diets showed reduced parasite invasion and development. Short-term feeding of diets high in n-3FA to young chicks may be a practical method for reducing production losses from cecal coccidiosis.

Effects of formaldehyde fumigation of housing on carotenoid pigmentation in three breeds of chickens.

Three breeds of chickens (two broiler breeds, A and B, and one layer breed, C) were raised from 1 day to 3 wk of age in either standard housing or housing additionally fumigated with formaldehyde. Groups of chicks from each breed in both housings received either regular poultry starter ration or this ration supplemented with marigold meal. At 3 wk of age, chicks of all three breeds had higher (111, 113, and 115%) mean body weights when raised in fumigated housing. There was no significant effect of supplemental carotenoids on body weight. Chicks of all three breeds raised in fumigated housing had significantly higher plasma carotenoid concentrations when fed either regular or supplemented feed, and, with the exception of Breed A on regular feed, also had significantly increased skin carotenoid values. No significant differences among breeds in plasma or skin carotenoid values were seen in chicks fed regular ration. However, significant differences were seen among groups of chicks fed supplemental carotenoids. The layer breed, C, had the highest percentage increase in carotenoid pigmentation when fed the carotenoid-supplemented diet in fumigated housing.

Effect of virginiamycin on serum carotenoid levels and long, segmented, filamentous organisms in broiler chicks.

Virginiamycin (Stafac 20) was mixed with feed at three levels recommended for chickens--5.5, 11, and 22 mg/kg (respectively 5, 10 and 20 g/ton)--and fed to broiler chicks. When fed from 1 day through 2 weeks of age, the drug appeared to retard infection of the lower small intestine by long, segmented, filamentous organisms (LSFOs), and at 2 weeks of age serum carotenoids in treated chicks were significantly higher than levels found in unmedicated chicks. However, as chicks were grown out to 4 and 6 weeks of age, the drug did not completely prevent eventual LSFO infection, even at the highest dose, and mean serum carotenoid levels in treated chicks were not significantly different from levels in control chicks. However, chicks fed at the level of 22 mg/kg had fewer LSFOs. Withdrawal of virginiamycin from treated chicks at 4 weeks of age allowed LSFO infection to occur but did not significantly affect serum carotenoid levels. When the drug was fed for 2 weeks at 22 mg/kg to 2-week-old chicks already infected with LSFOs, the bacteria could no longer be detected, suggesting that virginiamycin may aid the natural decline in LSFO population. Coincident with this treatment, serum carotenoids were higher, but not significantly so. Virginiamycin did not significantly increase the mean body weights of chicks in any of these experiments.

Effect of coccidiosis on the distribution of dietary lutein in the chick.

A series of experiments was run to determine whether the distribution of dietary carotenoids in chick plasma and tissue was dependent upon a saturable carrier, and to observe how infection with Eimeria acervulina affected this distribution. Broiler (Hubbard x Hubbard) and Sex Sal (Rhode Island Red x White Rock) chicks were fed a white corn and soybean meal-based diet supplemented with varying levels of a commercial lutein concentrate. Plasma, small intestine, and liver were analyzed for lutein. In the first experiment, mean plasma values in control broiler chicks increased linearly from 0 to 200 ppm supplementation. Mean plasma values of infected chicks were decreased an average of 71% as compared with controls, and reached a plateau between 100 and 200 ppm dietary lutein. In the second experiment, plasma, small intestinal, and liver lutein levels in broiler chicks reached a plateau between 200 and 500 ppm. In the third experiment, plasma and liver values in Sex Sal chicks leveled off between 250 and 500 ppm whereas levels in the small intestine increased linearly. Infection of the Sex Sal chicks at 3 wk of age with E. acervulina caused plasma lutein levels to be reduced about 88%, small intestinal levels 76%, and liver levels 72%. Supplementation with 500 ppm resulted in higher proportions of lutein in tissues. Infection with E. acervulina shifted the ratios to even higher proportions in the tissues at all supplementation levels.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of dietary cholesterol on the lipoprotein profile and binding of radioiodinated lipoproteins to hepatic membranes in the cockerel (Gallus domesticus).

1. Cockerels fed a cholesterol-supplemented diet experienced a marked elevation of lipoprotein particles of density less than or equal to 1.006 g/ml (VLDL) and a diminution of lipoprotein particles of density 1.02-1.05 g/ml (LDL). 2. Unlike VLDL of some cholesterol-fed animals, cholesterol-fed cockerel VLDL did not display beta-mobility on agarose gel electrophoresis. 3. [125I]LDL and [125I]HDL binding to cockerel liver membranes was not affected by cholesterol feeding. 4. Different lipoprotein types appear to bind to a common site on cockerel liver membranes. 5. The results suggest that liver cells of cockerels may not possess LDL binding sites that are analogous to those of mammalian species.