Dietary inclusion of different multi-strain complex probiotics; effects on performance in broilers

1. This study was designed to evaluate the effects of two different complex probiotic supplementations on the growth performance, meat quality, excreta microflora, nutrient retention, blood metabolic profile and noxious gas emissions in broilers.

2. A total of 612 conventional healthy 1-d-old Ross 308 broilers with body weight of 41 ± 0.3 g were randomly divided into 3 treatments with 12 replicate cages, with 17 broilers in each cage and fed with the following diets: CON-Basal diet, T1-CON + Probiotic A, T2-CON + Probiotic B.

3. Significant results were observed on body weight gain, but not on feed conversation ratio and feed intake, in the whole experimental period. Increased faecal lactobacillus counts were found with probiotics supplementation. However, no significant effects were found for meat quality, nutrient retention, blood metabolic profile or noxious gas emissions.

4. In conclusion, both multi-strain probiotics had beneficial effects on growth performance, drip loss percentage and faecal Lactobacillus counts in broilers.

     

Polymorphism identification and cardiac gene expression analysis of the calsequestrin 2 gene in broiler chickens with sudden death syndrome

1. Sudden death syndrome (SDS) in broilers is a cardiac disease associated with ventricular tachycardia (VT) and ventricular fibrillation (VF); however, its pathogenesis at the molecular level is not precisely determined.

2. Downregulation and mutations of calsequestrin 2 (CASQ2), a major intracellular Ca²⁺ buffer, have been associated with VT and sudden cardiac death (SCD) in humans but in chickens there is no report describing CASQ2 abnormalities in cardiac diseases.

3. In order to better understand the molecular mechanisms predisposing the myocardium to fatal arrhythmia in broilers, the mRNA expression level of chicken CASQ2 gene (chCASQ2) in the left ventricle of dead broilers with SDS was determined and compared to healthy broilers using quantitative real-time PCR (qPCR). To determine the probable mutations in chCASQ2, PCR and direct sequencing were also done.

4. Results showed a reduction in chCASQ2 expression in broilers dead by SDS. Three novel mutations (K289R, P308S, D310H) which are absent in healthy broilers were observed in chCASQ2.

5. It is concluded that susceptibility to fatal cardiac arrhythmia in SDS may be associated with changes in intracellular Ca²⁺ balance due to mutation and downregulation of chCASQ2.

     

Effects of ionised or chelated water-soluble mineral mixture supplementation on growth performance,nutrient digestibility,blood characteristics,meat quality and intestinal microbiota in broilers

1. An experiment was conducted to study the effects of dietary supplementation of water-soluble ionised or chelated mineral mixture on growth performance, nutrient digestibility, blood characteristics, relative organ weight, meat quality and excreta microflora in broilers.

2. A total of 408 Arbor Acres broilers (17 birds in 8 replicate pens) were randomly allocated into one of the following three treatments: (1) Control/basal diet (CON), (2) T1 (basal diet + 0.5% ionised mineral mixture solution, pH 3.0) and (3) T2 (basal diet + 0.5% chelated mineral mixture solution, pH 3.0).

3. The body weight gain was greater and feed conversion ratio was lower in broilers supplemented with ionised or chelated mineral liquid complex compared to CON during the grower and overall phase of the experiment.

4. No significant effect in the concentration of Ca and P in the blood was observed in birds supplemented with ionised or chelated mineral mixture solution. No adverse effects were observed in organ weight and meat quality with ionised or chelated mineral mixture supplementation.

5. Regarding intestinal microbiota counts there was a reduction of Escherichia coli counts in the small intestine in ionised mineral supplemented birds. In the large intestine, E. coli as well as Salmonella populations were reduced in ionised mineral supplemented birds.

6. In conclusion, ionised or chelated minerals have partial positive effects in improving growth performance and reducing pathogenic bacteria load in the gastro-intestinal tract.

     

Effect of supplementing broiler chicken diets with green tea extract on the growth performance,lipid profile,antioxidant status and immune response

1. A trial was conducted to explore the effect of feeding graded concentrations of green tea extract (GTE; 125, 250, 500, 1000 and 2000 mg/kg) on the growth performance, serum lipid profile, liver glutathione-reduced, thigh muscle malondialdehyde and humoral immune response against Newcastle disease virus vaccines of broiler chickens from hatching to 42 d of age. The results of broilers fed on GTE were contrasted to those fed on butylated hydroxytoluene (BHT; 125 mg/kg) or unsupplemented controls.

2. The obtained results revealed no significant difference among treatments in the measured growth performance parameters (body weight, average daily gain, average daily feed intake and feed conversion ratio), per cent liveability and serum lipid profile (total lipid, total cholesterol, high-density, low-density and very low-density lipoprotein cholesterols).

3. Dietary intake of GTE resulted in a significant increase in the liver glutathione-reduced level compared to the control. The glutathione-reduced level of broilers fed on the BHT diet was not significantly different from that of broilers fed on GTE or control diets. Feeding GTE or BHT resulted in a significant decrease in the malondialdehyde level of meat tissue.

4. The specific antibody titre against Newcastle disease virus vaccines was significantly increased at 28 and 35 d of age in broilers fed on diets supplemented with GTE but not with BHT.

5. In conclusion, GTE was found to possess antioxidant and immunostimulant characteristics for broilers when supplemented to their diets, and its optimum inclusion level ranged from 125 to 500 mg/kg.

     

The role of bentonite binders in single or concomitant mycotoxin contamination of chicken diets

1. Concomitant presence of mycotoxins is more likely to appear than a single mycotoxicosis since many mycotoxigenic fungi grow and produce their toxic metabolites under similar conditions.

2. The present study was designed to evaluate the efficacy of 4 mycotoxin binders to protect meat-type chickens against single and concomitant administration in the feed of two mycotoxins, namely aflatoxin B1 (AFB1) and ochratoxin A (OTA) both at concentration of 0.1 mg/kg.

3. A total of 440 as hatched, d-old, Ross 308 broilers were reared for 42 d. There were 11 dietary treatments. Chickens were fed on either an uncontaminated basal diet, basal diet and AFB1, basal with concomitant presence of AFB1 and OTA, basal diet and three binders A, B and C (1%) with or without AFB1 or basal diet and binder D (0.5%) with or without concomitant presence of AFB1 and OTA. Performance, carcass yield and several biochemical parameters were examined.

4. Mycotoxin concentration in liver and breast muscle samples was determined. Broiler performance under concomitant mycotoxin contamination was poorer than that under single mycotoxicosis. Mycotoxin presence increased relative heart weight compared to that of broilers fed on uncontaminated diets. Only OTA and not AFB1 was detected and only in the liver. OTA concentration was four-fold lower in broilers fed on a diet with binder compared to those fed on contaminated diets without binder.

5. In conclusion, the study revealed that binder composition and presence or not of multiple toxins may be important factors for optimum broiler performance under mycotoxicosis.

     

Protective effects of Urtica dioica seed extract in aflatoxicosis: histopathological and biochemical findings

1. The ameliorative potential and antioxidant capacity of an extract of Urtica dioica seeds (UDS) was investigated using histopathological changes in liver and kidney, measuring serum marker enzymes, antioxidant defence systems and lipid peroxidation (malondialdehyde (MDA)) content in various tissues of broilers exposed to aflatoxin (AF).

2. A total of 32 broilers were divided randomly into 4 groups: control, UDS extract-treated, AF-treated and AF+UDS extract-treated. Broilers in control and UDS extract-treated groups were fed on a diet without AF. The AF-treated group and AF+UDS extract-treated groups were treated with an estimated 1 mg total AF/kg feed. The AF+UDS extract groups received in addition 30 ml UDS extract/kg diet for 21 d.

3. The AF-treated group had significantly decreased body weight gain when compared to the other groups.

4. Biochemical analysis showed a small increase in the concentrations of serum aspartate aminotransferase, alanine aminotransferase, gamma glutamyl transpeptidase and lactate dehydrogenase in the AF-treated group compared to that of the control group, whereas concentrations of these enzymes were decreased in the AF+UDS group compared to that of the AF-treated group.

5. Administration of supplementary UDS extract helped restore the AF-induced increase in MDA and reduced the antioxidant system towards normality, particularly in the liver, brain, kidney and heart. Hepatorenal protection by UDS extracts was further supported by the almost normal histology in AF+UDS extract-treated group as compared to the degenerative changes in the AF-treated broilers.

6. It was concluded that UDS extract has a protective hepatorenal effect in broilers affected by aflatoxicosis, probably acting by promoting the antioxidative defence systems.

     

Yellow mealworm larvae (Tenebrio molitor,L.) as a possible alternative to soybean meal in broiler diets

1. The aim of the study was to evaluate the feasibility of replacing soybean meal (SBM) with Tenebrio molitor larvae (TML) meal in broiler diets.

2. A total of 80 30-d-old male Shaver brown broilers were divided into two groups fed on two isoproteic and isoenergetic diets differing for protein source (SBM vs. TML).

3. Up to 62 d of age, body weight and feed intake were recorded weekly and body weight gain, feed conversion ratio (FCR), protein efficiency ratio (PER) and European efficiency factor (EEF) were calculated. At 62 d, blood samples were collected from 16 birds/group for evaluation of blood profiles.

4. Feed intake was not different between groups considering the entire period of the trial. The FCR was more favourable in the TML than SBM group from 46 d of age and in the entire period of the trial (4.13 vs. 3.62). The PER was higher in the SBM than in the TML group (1.92 vs. 1.37) while the EEF was higher in broilers fed on the TML diet (132.6 vs. 156.2).

5. Albumin-to-globulin ratio was higher in broilers fed on SBM than in the other group (0.44 vs. 0.30). aspartate aminotransferase and alanine aminotransferase were higher in TML than SBM (195.1 vs. 178.6 U/l and 82.07 vs. 46.71 U/l, respectively). Uric acid was higher in broilers fed on SBM than TML (5.40 vs. 4.16 mg/dl).

6. TML did not affect feed intake and growth rate of broilers from 30 to 62 d of age when compared to an isoproteic and isoenergetic SBM diet, but FCR of the TML group was more favourable than that of the SBM group. The lowest albumin-to-globulin ratio in broilers fed on TML suggests a higher immune response, probably due to the prebiotic effects of chitin.

     

The influence of feeding crimped kernel maize silage on broiler production,nutrient digestibility and meat quality

1. Two experiments were carried out in parallel with male Ross 308 broilers over 37 d. An experiment with a total of 736 broilers was performed to study the effect of dietary inclusion of crimped kernel maize silage (CKMS) on broiler production and meat quality. Another study with 32 broilers was carried out from 21 to 25 d to investigate the inclusion of CKMS on nutrient digestibility.

2. In both trials, 4 dietary treatments were used: wheat-based feed (WBF), maize-based feed (MBF), maize-based feed supplemented with 15% CKMS (CKMS-15) and maize-based feed supplemented with 30% CKMS (CKMS-30).

3. Compared with MBF, the dry matter (DM) intakes of broilers receiving CKMS-15 and CKMS-30, respectively, were numerically 7.5 and 6.2% higher and feed conversion ratio 6 and 12% poorer (significant for 30% CKMS), although there were no significant differences in AME content between the three diets. At 37 d, the body weight of birds receiving 15% CKMS was similar to birds fed with MBF. However, the inclusion of 30% CKMS decreased broiler growth. Dietary supplementation with CKMS significantly reduced the apparent digestibility of phosphorus. The fat digestibility was significantly lower for CKMS-30 than for the other three diets.

4. Broiler mortality decreased significantly when CKMS was added to the diet.

5. The consumption of drinking water was significantly lower in all maize-based diets as compared to WBF and was lowest in broilers fed with CKMS-30.

6. An improved litter quality in terms of DM content and a lower frequency of foot pad lesions was observed with broilers supplemented with both dietary levels of CKMS.

7. The addition of CKMS to maize-based diets increased juiciness, tenderness and crumbliness of the meat.

8. In conclusion, the dietary supplementation of 15% CKMS had no negative effect on broiler growth and positively influenced bird welfare in terms of mortality and foot pad health. Therefore, the addition of 15% CKMS to maize-based diets is considered an advantageous feeding strategy in broiler production.

     

Natural zeolites in diet or litter of broilers

1. This study aims to analyse the influence of adding natural zeolites (clinoptilolite) to the diet or litter of broilers and their effects on growth performance, carcass yield and litter quality.

2. Three consecutive flocks of broilers were raised on the same sawdust litter, from d 1 to d 42 of age, and distributed in three treatments (control with no added zeolites, addition of 5 g/kg zeolite to diet and addition of 100 g/kg zeolites to litter).

3. The addition of zeolites to the diet or litter did not affect growth performance or carcass yield.

4. The addition of zeolites to the diet did not influence moisture content of the litter, ammonia volatilisation was reduced only in the first flock and pH of litter was reduced in the second and third flock.

5. However, the addition of zeolites to the litter reduced moisture content, litter pH and ammonia volatilisation in all flocks analysed.

6. The addition of 5 g/kg zeolite to the diet in three consecutive flocks was not effective in maintaining litter quality, whereas the addition of 100 g/kg natural zeolites to sawdust litter reduced litter moisture and ammonia volatilisation in three consecutive flocks raised on the same litter.

     

Haematological parameters,serum lipid profile,liver function and fatty acid profile of broiler chickens fed on diets supplemented with pomegranate seed oil and linseed oil

1. The objective of the present study was to determine effect of pomegranate seed oil (PSO) and linseed oil (LO) on haematological parameters, serum lipid profile and liver enzymes as well as fatty acids profile of adipose tissue in broilers.

2. Broilers (n = 400) were fed on diets containing graded PSO levels (0.0%, 0.5%, 1.0%, 1.5%) with or without 2% LO. After 6 weeks of feeding, 6 male broilers from each group were slaughtered and abdominal fat, liver and blood samples were collected.

3. Mixtures of pomegranate seed oil (0.5%, 1%) with linseed oil increased white blood cell level in broilers. Total cholesterol was elevated after LO supplementation whereas administration of PSO (1.5%) significantly decreased this parameter. PSO administration caused c9,t11 conjugated linoleic acid (CLA) concentration-dependent deposition in adipose tissue. By LO addition α-linolenic acid (ALA) content was enhanced, decreasing the n-6/n-3 ratio. PSO and ALA also affected oleic acid proportion in adipose tissue. Neither pomegranate seed oil nor linseed oil had any effect on liver parameters.

4. Pomegranate seed oil had no negative effects on broiler health status and can be considered as a functional poultry meat component.

     

The normalisation of CAPN gene expression in M. pectoralis superficialis in broiler lines differing in growth rate and their relationship to breast muscle tenderness

1. The aim of this study was to assess mRNA abundance of calpain 1 (CAPN1) and calpain 3 (CAPN3) in breast muscle of 80 fast-growing (FG) and slow-growing broilers (SG) and relate gene expression in relation to growth and Warner Bratzler (WB) shear force of breast muscle.

2. The expression of CAPN1 and CAPN3 genes was higher in the FG compared to the SG line, but significant results were obtained only for CAPN1. The CAPN1 mRNA level was strongly dependent on line and gender interaction.

3. Lower values of shear force were observed in the FG line, where a higher level of calpain expression was shown.

4. A new panel of housekeeping genes (RPL4 and SDHA) for normalisation of gene expression in muscle tissues could be used in other studies of gene expression in chicken.

     

Influence of an organic acid blend and essential oil blend,individually or in combination,on growth performance,carcass parameters,apparent digestibility,intestinal microflora and intestinal morphology of broilers

1. This study aimed to evaluate the influence of an organic acid (OA) and essential oil (EO) blends, individually or in combination, on growth performance, carcass parameters, apparent digestibility, intestinal microflora and intestinal morphology of broilers.

2. A total of 480 one-d-old male Ross 308 chicks were randomly assigned to 4 treatments consisting of 4 replicates each (n = 30 birds in each replicate). Dietary treatments consisted of a basal diet (control), and basal diet supplemented with 2 g/kg OA blend (OAB), 300 mg/kg EO blend (EOB), or with 2 g/kg OA and 300 mg/kg EO blend (OAB–EOB) for 42 d.

3. The dietary supplementation with EO blend or in combination with OA blend increased body weight gain and improved feed efficiency as compared to control. Dietary treatments had no significant effects on feed consumption or relative organ weights of broilers.

4. The OAB diet increased carcass yield compared to the control diet but the lowest carcass yield occurred with the OAB–EOB combination. Birds fed on EOB and OAB–EOB diets had lower ileum Escherichia coli counts than birds fed on the control diet.

5. There was no significant effect of treatments on apparent digestibility at 16–21 d but the EOB and OAB–EOB diets increased apparent digestibility of dry matter and crude protein during the finisher period (d 37–42) compared to the control diet.

6. Birds fed on the EOB and OAB–EOB diets had greater villus height in the ileum at 21 and 42 d of age and had lower crypt depth in the ileum at 42 d of age than birds given the control diet.

7. In conclusion, beneficial effects of the use of EO blend individually or in combination with the OA blend were observed but the OA blend alone was ineffective. Furthermore, the use of the combination of OA and EO was more effective, in some respects, than their individual use.

     

Lactic acid bacteria isolated from poultry protect the intestinal epithelial cells of chickens from in vitro wheat germ agglutinin-induced cytotoxicity

1. Poultry fed on wheat-based diets regularly ingest wheat germ agglutinin (WGA) that has toxic effects in vitro on intestinal epithelial cells (IEC) obtained from 14-d-old broilers. Cytotoxicity and the potential role of 14 intestinal bacterial strains in the removal of bound lectins in epithelial cell cultures were investigated.

2. Cytotoxicity was dependent on time and lectin concentration; the lethal dose (LD₅₀) was 8.36 µg/ml for IEC exposed for 2 h to WGA.

3. Complementary sugars to WGA were detected on the surface of one Enterococcus and 9 Lactobacillus strains isolated from poultry. These strains were evaluated as a lectin removal tool for cytotoxicity prevention.

4. Incubation of lactic acid bacteria with WGA before IEC–lectin interaction caused a substantial reduction in the percentage of cell deaths. The protection was attributed to the amount of lectin bound to the bacterial surfaces and was strain-dependent. L. salivarius LET 201 and L. reuteri LET 210 were more efficient than the other lactic acid bacteria assayed.

5. These results provide a basis for the development of probiotic supplements or cell-wall preparations of selected lactic acid bacteria intended to avoid harmful effects of a natural constituent of the grain in wheat-based diets.

     

Silicon in broiler drinking water promotes bone development in broiler chickens

1. Skeletal abnormalities, bone deformities and fractures cause significant losses in broiler production during both rearing and processing. Silicon is an essential mineral for bone and connective tissue synthesis and for calcium absorption during the early stages of bone formation.

2. Performance was not affected by the addition of silicon. However, broilers receiving silicon showed a significant increase of phosphorus, zinc, copper, manganese and ash in the tibia.

3. In conclusion, broiler performance was not impaired by adding the tested silicon product to the drinking water. In addition, bone development improved, as demonstrated by higher mineral and ash content.

4. Further studies are required to determine the optimal concentration of silicon, including heat stress simulations, to better understand the effects of silicon on bone development.

     

Changes in eggshell structure and predisposition of broilers to health problems: is there a common pathophysiology?

1. A study of broiler breeder eggs differing in eggshell matrix optical density was conducted to determine the association of eggshell structural quality and the risk of disease in broilers.

2. A total of 10 000 eggs from a broiler breeder flock were examined according to the pre-established criteria, and allocated to groups classified as having a high or low density shell matrix.

3. The eggs from respective groups were incubated and hatched in a commercial hatchery. Samples of unhatched eggs from each group were subjected to detailed examination to establish the cause of reproductive failure. First, quality chicks from each group were raised as separate flocks in a commercial broiler barn. Group performance, morbidity, and mortality were monitored throughout the growth period. All birds were processed in a commercial plant, and condemnation data were compiled.

4. There were significantly more unhatched eggs, and fewer quality chicks in the group classified as having a low density eggshell matrix, in comparison to the high-density group.

5. Embryo pathology accounted for a large proportion of the overall reproductive failure in both groups, with a large proportion of embryos showing anatomical anomalies. The eggs and embryos from the low-density eggshell matrix group were three times more likely to be infected.

6. Significantly higher production losses associated with mortality/morbidity, and condemnations of carcasses at processing were observed in broilers from the low-density eggshell group compared with the high-density eggshell group.

7. It is concluded that common metabolic/physiological changes in breeder hens associated with eggshell pathology may be also risk factors linked with predisposition of broiler chicks to some health problems.

     

Performance and carbon turnover in fast- and slow-growing broilers submitted to cyclic heat stress and fed on high-protein diets

1. Two experiments were conducted to test the hypothesis that when using similar protein/amino acid diets and environment temperature conditions, the performance and carbon turnover in muscle and liver tissues, as measured by the incorporation of stable isotopes (¹³C/¹²C), must be different between fast-growing Cobb 500® and slow-growing Label Rouge broilers.

2. For both experiments (Cobb and Label Rouge), 21-d-old birds were distributed in a completely randomised, 3 × 3 factorial design; three environmental temperatures (cyclic heat stress ad libitum, 22°C ad libitum, and 22°C restricted) and three crude protein concentrations (189.1, 210 and 220 g/kg CP) were used.

3. The Cobb 500® had better performance with higher concentrations of crude protein. Cyclic heat stress (a temperature factor), negatively affected this genetic strain’s performance. For the Label Rouge birds, the crude protein concentrations in the diet presented inconsistent results and cyclic heat stress did not affect the performance.

4. The carbon turnover rate was affected in the Cobb 500® strain, with a high protein content reducing carbon turnover in the evaluated tissues (liver and muscles). Feed intake had a greater impact on carbon turnover rates than cyclic heat stress. The Label Rouge birds were not affected by the evaluated factors, suggesting that genetic improvement has a leading role on tissue carbon turnover.

5. There is a genetic influence on carbon turnover in the liver and muscle tissues of broiler chickens. In addition, genetically fast-growing broilers are more susceptible to variations in diet composition and environmental temperature than less rapidly growing animals.

     

Comparative effects of zinc-nano complexes,zinc-sulphate and zinc-methionine on performance in broiler chickens

1. Micronutrients, especially zinc, have an important role in normal metabolism and growth of broilers. Using novel technologies helps to synthesise novel zinc complexes to deliver this micronutrient more efficiently.

2. In the present study, the effects of different zinc complexes and nano complexes on broiler performance were compared. Broilers in 6 groups were given basal diet (without zinc) and basal diet supplemented with zinc-sulphate, zinc-methionine, zinc-nano-sulphate, zinc-nano-methionine and zinc-nano-max (that was synthesised based on nanochelating technology) at a concentration of 80 mg/kg of diet.

3. At 1–42 d of age, dietary zinc-nano-sulphate supplementation decreased weight gain and feed intake. However, feed conversion ratio was not influenced by treatments. Carcass yield (%) of birds in the zinc-nano-sulphate and control groups were dramatically reduced at 42 d of age and abdominal fat (%) increased in these groups.

4. Relative to the control group, the antibody titre, spleen and bursa of Fabricius (%) were significantly higher in groups supplemented with zinc. Heterophil (%) was also significantly higher in the zinc-nano-methionine group in blood on d 42 compared to the control, zinc-sulphate and zinc-nano-sulphate.

5. Compared to the controls, the mean malondialdehyde content in thigh tissue was significantly reduced in groups supplemented with zinc at the time 0, 50, 100 and 150 min after oxidation.

6. Tibia zinc concentration in nanoparticle zinc samples was significantly higher relative to the control and zinc-sulphate groups.

7. Taken together, our data indicate that delivery of zinc in the structure of zinc-nano-methionine and zinc-nano-max at concentrations of 80 mg/kg of diet improves growth performance. However, dietary zinc-nano-sulphate decreased growth performance in broilers.

     

Accelerated digestion of nucleic acids by pepsin from the stomach of chicken

1. Nucleic acids have become an important nutritional supplement in poultry feed; however, the digestion of nucleic acids in poultry is unclear. The objective of this study was to investigate the digestion of nucleic acids by chicken pepsin in vitro.

2. The extracted pepsinogen from the stomach of the chicken was purified to homogeneity. Upon activation at pH 2.0, chicken pepsinogen was converted to its active form.

3. Nucleic acids, including λ-DNA, salmon sperm DNA and single-strand DNA (ssDNA), can be used as substrates and digested into short-chain oligonucleotides by pepsin.

4. Interestingly, the digestion of the nucleic acids was inhibited when pepsin was treated by alkaline solution (pH 8.0) or pepstatin A. Also, the digestion of the nucleic acids was not affected by the addition of haemoglobin or bovine serum albumin.

5. The results suggested that nucleic acids could be digested by chicken pepsin. Thus pepsin may have a role in digesting nucleic acids in vivo. Nucleic acids added to poultry fed may be digested, starting from the stomach.

     

Increased lipid peroxidation occurs during development in Japanese quail (Coturnix japonica) embryos

1. The growth of the avian embryo is paralleled by an exponential increase in the rate of whole-embryonic oxygen consumption, which potentially increases oxidative damage.

2. Age-related patterns of tissue lipid peroxidation were characterised in brain, liver and heart tissue of developing Japanese quail (Coturnix japonica) embryos between 9 and 15 d of age, over which embryo mass increased by a factor of 6. Lipid peroxidation was quantified in each tissue by spectrophotometric measurement of malondialdehyde using the thiobarbituric acid reactive substances assay.

3. In all tissues, lipid peroxidation increased greatly as development proceeded. Concentrations of malondialdehyde increased in parallel with the cumulative amount of oxygen consumed by the developing embryo consistent with the hypothesis that oxidative stress results from the production of free radicals due to oxidative metabolism.

4. This study describes in vivo oxidative stress in developing avian embryos and suggests that rates of embryonic growth, oxidative metabolism and oxidative damage likely vary in parallel.