Management and biosecurity practices by small to medium egg producers in Scotland.

1. Information about procedures and biosecurity practices used by small and medium egg producers (SMEPs) is scarce. Anecdotal evidence suggests that biosecurity in such enterprises may be poor, as personnel and equipment move freely between sites and this may be compounded by personnel working on commercial units who keep their own poultry.2. To fill this knowledge gap, a questionnaire was designed and implemented targeting SMEPs in Scotland. Small enterprises were defined as egg producers that have ≥50 laying hens but <350 laying hens; while medium enterprises were defined as egg producers that have ≥350 laying hens but ≤32 000 laying hens. The questionnaire consisted of a total of 56 questions divided into multiple sections, covering the characteristics of the primary keeper, location of the enterprise and size of the flocks, husbandry, marketing of products and health/biosecurity.3. The questionnaire was posted to 375 holdings at the beginning of March 2017 and the survey remained open until the end of May 2017. In total 90 questionnaires were received by the cut-off date of which 76 questionnaires were from SMEPs. Forty were small enterprises and 36 were medium enterprises. For three questionnaires, it was not possible to identify the enterprise type.4. Differences were observed between SMEPs in terms of reported biosecurity and management practices, with medium enterprises reporting the adoption of more biosecurity measures than small enterprises. Furthermore, SMEPs behave differently from backyard poultry keepers and large commercial companies in terms of disease risk.5. In conclusion, it is important to ensure that SMEPs are considered in contingency plans and disease control programmes and that engagement with them is promoted so that the uptake of relevant information, such as awareness of disease control programmes, is optimised.

Reliably colonising broiler chickens with Campylobacter spp. using a litter-based method.

1. Chicken-associated Campylobacter spp. are the cause of most food poisoning cases in Europe. In order to study the host-pathogen interactions, a reliable and reproducible method of colonising chickens with the bacteria is required. 2. This study aimed to identify a more appropriate and less invasive method of colonisation (cf. gavaging) by seeding bedding material (litter) that commercial chickens are kept on with a mixture of Campylobacter spp., broth and faeces. 3. The first phase of the study tested the longevity of Campylobacter spp. recovery in seeded litter over 24 h: significantly more Campylobacter spp. was recovered at 0 or 3 h post-seeding than at 6 and 24 h post-seeding, indicating that the pathogen can survive to detectable levels for at least 3 h in this environment. 4. In the second phase, three groups of 10 broiler chickens (negative for Campylobacter spp. prior to exposure) were exposed at 21 days of age to one of three different Campylobacter jejuni and C. coli mixes (A, B, C), using the method above. At 28 days of age, birds were euthanised by overdose of barbiturate or cervical dislocation, and livers and caeca removed for Campylobacter spp. assessment. 5. All liver and 28/30 caeca samples tested positive for Campylobacter spp., with mix A and C giving higher counts in the caeca than mix B. The method of euthanasia did not affect Campylobacter spp. counts. 6. In conclusion, a successful method for reliably colonising broiler chickens with Campylobacter spp. has been developed which negates the need for gavaging and is more representative of how contamination occurs in the field.

Early experiences matter: a review of the effects of prenatal environment on offspring characteristics in poultry.

Early life experiences can be important in determining offspring phenotypes and may influence interaction with the environment and hence health, welfare, and productivity. The prenatal environment of poultry can be divided into the pre-lay environment and the egg storage/incubation environment, both of which can affect offspring outcomes. The ability to separate maternal and egg/incubation effects makes birds well suited to this type of research. There are many factors, including feeding and nutrition, environmental conditions, husbandry practices, housing system, social environment, infectious environment, and maternal health status, that can influence both the health and performance and behavior and cognition of the offspring. There are some aspects of the environments that can be changed to produce beneficial effects in the offspring, like addition of certain additives to feed or short changes in incubation temperatures, while other aspects should be avoided to reduce negative effects, such as unpredictable feeding and lighting regimens. Measures of offspring characteristics may prove to be a useful method of assessing parent stock welfare if known stressors result in predictable offspring outcomes. This has the advantage of assessing the parent environment without interfering with the animals and possibly affecting their responses and could lead to improved welfare for the animals.

Effects of natural blend of essential oil on growth performance, blood biochemistry, cecal morphology, and carcass quality of broiler chickens.

The study evaluated the effect of a novel commercial preparation of natural blend of essential oils from basil, caraway, laurel, lemon, oregano, sage, tea, and thyme (Tecnaroma Herbal Mix PL) on growth performance, blood biochemistry, cecal morphology, and carcass quality of broilers. Six nutritionally adequate wheat and soybean-based diets were generated by the addition of Tecnaroma Herbal Mix PL at 0, 100, 200, 300, 400, and 500 g/t of feed. The diets were fed as crumbs in the starter phase (d 0-10) and as pellets during the grower (d 10-24) and finisher (d 24-42) phases. Nine hundred sixty 1-d-old chicks were allocated to the 6 dietary treatments each having 8 replicate pens with 20 birds per pen. The data obtained were analyzed using ANOVA with a P < 0.05 level of significance. Birds fed diets supplemented with Tecnaroma Herbal Mix PL had significantly heavier BW and higher (P < 0.05) weight gain and had improved (P < 0.05) feed to gain ratio compared with the control group during grower phase and overall performance. The blood biochemistry results showed no differences (P > 0.05) between treatments. The carcass weight, breast weight, and relative percentage of breast meat increased (P < 0.05) when diets were supplemented with Tecnaroma Herbal Mix PL compared with that from birds fed the control diet. The inclusion level of 300 g of Tecnaroma Herbal Mix PL/t of feed was optimum for enhancing breast meat yield and nutrient utilization as indicated by increased (P < 0.05) cecal villus surface area.

Persistent environmental reservoirs on farms as risk factors for Campylobacter in commercial poultry.

Campylobacter is the most common known source of human bacterial enteritis in the developed world and poultry is considered the main source. Broilers often become colonized with Campylobacter during rearing, and then contaminate the farm environment. The objective of this study was to identify Campylobacter-positive environmental reservoirs on farms, as these pose a risk to broiler flocks becoming colonized with Campylobacter. We considered the temporal aspects of exposure and colonization. A longitudinal study monitored six conventional rearing farms over 2 years. The broiler flocks, catchers' equipment, vehicles, shed surrounds, shed entrance, other equipment, farm entrance, other animals, puddles, dead birds, mains water and drinkers were systematically sampled 2-4 times per flock. A multivariable generalized estimating equation model was used to assess associations between contaminated environmental sites and colonized broiler flocks. The associations were adjusted for confounders and other known risk factors. To further assess temporality of contamination, the sequence of contamination of the different environmental sites and the flocks was established. Contaminated shed entrances and anterooms, contaminated drinkers and shedding of Campylobacter by other animals such as cattle, dogs, wildlife and rodents were significantly associated with positive flocks. The reservoir of 'other animals' was also the reservoir most commonly positive before the flock became colonized. The other sites usually became contaminated after the flock was colonized.

Physiological responses of laying hens during whole-house killing with carbon dioxide.

1. Poultry on farms are sometimes required to be killed in an emergency, such as during a disease epidemic, yet none of the available methods are ideal. Whole-house carbon dioxide (CO(2)) administration has practical advantages, but gives rise to welfare concerns. 2. The study measured the body temperature, respiration, cardiac and brain activity (EEG) responses of 10 adult hens placed in tiered cages in a deep pit house while the entire flock (28,000 end-of-lay hens) was killed with CO(2). Video and thermographic images were also recorded. Liquid CO(2) was injected into the building producing a gaseous concentration of 45% within 19 min. 3. Those hens nearest the gas delivery site showed delayed respiratory, cardiac and EEG responses compared with those at more distant locations. Although sub-zero temperatures were recorded in the immediate vicinity of some birds, body temperatures indicated that they did not die of hypothermia. 4. EEG characteristics strongly associated with unconsciousness were used to determine an unequivocal time to loss of consciousness; this ranged from 6·0 to 10·5 (average 7·8) min after onset of gas injection. Distinctive cardiac and respiratory responses were seen following gas exposure; in particular, birds responded to inhalation of CO(2) by deep breathing. 5. The primary welfare concern is the duration of unpleasant respiratory effects, such as deep breathing, while the birds were substantively conscious. However, the concentration of CO(2) to which the birds were exposed while conscious would not have stimulated nasal and oral nociceptors. Time to death varied between 12·0 and 22·1 min after gas delivery.

Providing laying hens with perches: fulfilling behavioural needs but causing injury?

1. The EU laying hen directive, which bans standard battery cages from 2012, has implications for animal welfare, particularly since housing laying hens in extensive systems, while increasing natural behaviour and improving bone strength, is associated with a greater level of bone fractures, predominantly of the keel bone, compared to birds housed in cages. 2. The aetiology and welfare consequences of keel and other bone fractures are not well understood and could have important implications for housing system designs. While proposed alterations to layer housing are based on the desire to fulfil behavioural needs and increase bone strength, there appears to have been little consideration of the effect of system on potential injury. 3. In addition, there are variations in how the directive is interpreted. For example, egg producers housing hens in extensive systems in Scotland and Northern Ireland must provide hens with aerial perches, whereas in England and Wales they do not. Aerial perches may be implicated in bone fracture injuries. 4. This paper reviews the prevalence of bone fractures in the egg-laying sector of the poultry industry and the literature on perches. It also explores how bone fractures may be occurring. 5. We propose some means of reducing bone fracture, namely through improved housing designs and genetic selection.

Sources and spread of thermophilic Campylobacter spp. during partial depopulation of broiler chicken flocks.

The practice of partial depopulation or thinning (early removal of a portion of birds from a commercial broiler flock) is a reported risk factor for Campylobacter colonization of residual birds because of the difficulty in maintaining biosecurity during the thinning process. The effect of this practice was studied in detail for 51 target flocks, each at a different growing farm belonging to one of seven major poultry companies throughout the United Kingdom. On 21 of these farms, the target flock was already colonized by Campylobacter, and at slaughter all cecal samples examined were positive, with a mean of 8 log CFU/g. An additional 27 flocks became positive within 2 to 6 days of the start of thinning and had similarly high levels of cecal carriage at slaughter. Just before the thinning process, Campylobacter was isolated frequently from the farm driveways, transport vehicles, equipment, and personnel. Strains from seven farms on which flocks became colonized after thinning were examined by pulsed-field gel electrophoresis typing. An association was found between strains occurring at specific sampling sites and those isolated subsequently from the thinned flocks. The results indicated that particular strains had spread from one farm to another when the farms were jointly owned by the same company and employed the same bird-catching teams and/or vehicles. These results highlight the need for better hygiene control in relation to catching equipment and personnel and more effective cleaning and disinfection of vehicles and bird-transport crates.

Socio-economic drivers for UK organic pullet rearers and the implications for poultry health.

1. Certified organic pullet producers were surveyed to gain a better understanding of the production environment, to identify the key constraints to organic pullet rearing and to identify factors that affected bird health. 2. Pullet rearers had been involved in organic production for between 1 and 12 years. 3. The number of pullets reared per annum ranged from 6 to 12 000 and the number of birds housed per unit from <50 to >1000. 4. The primary reason for being involved in organic production was given as 'commercial' with 'environmental' and 'welfare' being the next most popular categories. 5. Fewer than 50% of the respondents vaccinated their flocks and, for those that were protected, the diseases vaccinated against frequently were Newcastle disease, infectious bronchitis and Marek's disease. Annual mortality ranged from <2 to >7% with smothering accounting for 25% of all mortality. 6. Approximately 40% of respondents saw no constraints to rearing organic pullets while others identified a range of factors including capital, availability of land and inadequate margins as being the primary constraint.

Post-feeding activity of Lucilia sericata (Diptera: Calliphoridae) on common domestic indoor surfaces and its effect on development.

Developmental data of forensically important blowflies used by entomologists to estimate minimum post mortem interval (mPMI) are established under controlled laboratory conditions for various temperature ranges throughout the stages of egg, 1st-3rd instar, puparia, and adult fly emergence. However, environmental conditions may influence the patterns of development and behaviour of blowflies, potentially impacting on these established development rates. Previous studies investigating indoor colonisation have focused on the delay to oviposition, with behaviour during the post-feeding phase in this setting often overlooked. The environment in which third instar larvae disperse when searching for a pupariation site may vary drastically at both outdoor and indoor scenarios, influencing the activity and distance travelled during this phase and possibly affecting developmental rates. This study investigated the effect of eight common domestic indoor surfaces on dispersal time, distance travelled, and behaviour of post-feeding Lucilia sericata as well as any resulting variation in development. It was found that pupariation and puparia length within a pupariation medium of sawdust (often used in laboratory settings) produced comparable results with that of carpeted environments (those deemed to be 'enclosed'). Non-carpeted environments (those which were 'exposed') produced a delay to pupariation likely due to increased activity and energy expenditure in searching for pupariation sites which enabled burial. In addition, the observed speed of travel during dispersal was seen via time lapse photography to be greater within 'exposed' conditions. Larvae which dispersed upon burnt laminate flooring were observed to travel faster than in all other conditions and showed the only significant variation (P=0.04) in the day of emergence in comparison to the control condition of sawdust. This study has demonstrated that wandering phase activity is affected by the environmental surface which has potential implications for estimating both the distance travelled by dispersing larvae in indoor conditions and with further research, may be a consideration in mPMI calculations.

Effects of supplementing broiler breeder diets with organoselenium compounds and polyunsaturated fatty acids on hatchability.

The effects of supplementing broiler breeder diets with polyunsaturated fatty acids (PUFA) and organoselenium compounds on fertility, hatchability, and the weight of 1-d-old chicks was assessed. Prepeak (23 wk) and peak (27 wk) production breeders were fed 1 of 4 diets: a wheat-based commercial breeder diet with 55 g/kg of either soybean oil (SO) or fish oil (FO), but no added Se (only that originating from feed ingredients), and each diet with added Se as Sel-Plex (SO + Se, FO + Se). The diets were designed to contain <0.1 mg/kg of Se and about 0.5 mg/kg of Se for the nonsupplemented (no added Se) and the supplemented diets, respectively. The Se concentration of the eggshell of the hatching egg was measured. The concentration of Se, PUFA, and total lipid content of the brain and liver of the 1-d-old chick was determined. The number of fertile eggs increased, embryonic mortality decreased, and hatchability increased as hen age increased from 23 to 27 wk. The Se concentration in the eggshell and the brain and liver of 1-d-old chicks was higher in the high-Se treatments com pared with the concentration in the low-Se treatments. Fish oil inclusion in the breeder diet increased embryonic mortality in wk 3 of incubation and reduced both hatchability and 1-d-old chick weight in hens of both ages. The addition of Se to the FO diets ameliorated some of these adverse effects, because chicks hatched from eggs laid by 23-wk-old breeders of the FO + Se treatment were heavier than those receiving the FO treatment. The Se concentration in the brain and liver of chicks from the FO hens was higher than that in chicks from the SO hens. The concentration of docosahexaenoic fatty acid was higher in the liver of chicks from the SO + Se treatment compared with that of chicks from the SO treatment, indicating possible protective effects of Se. Hatchability was decreased by increased PUFA and was higher in 27-wk-old compared with 23-wk-old breeders.

Review of gaseous methods of killing poultry on-farm for disease control purposes.

Poultry may need to be culled in the event of an outbreak of disease. Gassing has advantages over mechanical and electrical methods or overdoses of anaesthetics because large numbers can be killed simultaneously and little or no handling of the birds is required. However, gaseous killing methods may have welfare implications for the birds, which may find various gases more or less aversive, may undergo respiratory distress and/or experience convulsions, and may remain conscious for a considerable time before they die. In addition, the gases used may present health and safety risks to human operators, and be difficult to supply and deliver.

Effects of supplementing broiler breeder diets with organic selenium and polyunsaturated fatty acids on egg quality during storage.

We assessed the effects of supplementation of broiler breeder diets with polyunsaturated fatty acids (PUFA) and organic Se on hatching egg quality during storage. Broiler breeders (n = 352) were fed 1 of 4 diets: SO (soybean oil with no added Se), SO with Se (SO+Se), fish oil (FO), or FO and Se (FO+Se). Eggs from prepeak (23 wk) and peak production (27 wk) breeders were stored for 14 d under typical (15 degrees C, 78% RH) conditions. Eggs were analyzed for albumen Haugh units (HU) and pH, shell characteristics, egg component weight, Se content, and fatty acid (FA) profile. The efficiency of transfer of Se into the egg was greater in low Se treatments compared with in high Se treatments. The majority of Se from the low Se treatments was deposited in the yolk, whereas in high Se treatments, the Se was deposited evenly in the yolk and albumen. In the low Se treatments, the Se content of yolk and albumen was lower than in the high Se treatments and decreased as age increased. Albumen HU decreased with increased storage, although high Se treatments had greater HU compared with low Se treatments. Eggs from the FO treatment were smaller with thinner eggshells compared with the SO treatment, but addition of Se to the FO diets ameliorated some of these effects. The egg yolk FA profile from the SO+Se treatment was altered compared with that from the SO treatment, perhaps due to interactions between dietary PUFA and Se-dependent enzyme systems. Hatching egg quality during storage was affected by dietary PUFA and Se content, as well as broiler breeder age.

Genome wide transcriptomic analysis identifies pathways affected by the infusion of Clostridium perfringens culture supernatant in the duodenum of broilers in situ.

Clostridium perfringens type A is the main etiological factor for necrotic enteritis, a multifactorial enteric disease that penalizes performance, health, and welfare of poultry. Lack of knowledge of host responses and disease pathogenesis is slowing down progress on developing therapies for disease control. A combined genomewide and targeted gene approach was used to investigate pathways and biological functions affected by the infusion of C. perfringens culture supernatant in the duodenum of broilers in 2 experiments. An in situ isolated loop of duodenum was prepared in anesthetized broilers of 3 wk of age (Exp. 1) and was infused either with crude C. perfringens culture supernatant (n = 7; treated), positive for necrotic enteritis B-like toxin (NetB) as determined by a cytotoxicity assay, or with a control preparation (n = 6; control). Birds were maintained alive for 1 h and then euthanized for tissue recovery. The use of the Affymetrix chicken genome array on RNA samples from loop tissue showed top biological functions affected by culture supernatant infusion included cell morphology, immune cell trafficking, and cell death; pathways affected included death receptor signaling, inflammatory response, and nuclear factor (NF)-κB signaling. In a second in situ study (Exp. 2), broilers were maintained alive for 4 h to monitor temporal expression patterns of targeted genes. Duodenal tissue was removed at 0.5, 1, 2, and 4 h post-infusion with culture supernatant (n = 9) or a control preparation (n = 5) for histology and gene expression analysis. Genes encoding proinflammatory cytokines, such as interferon γ (IFNγ), cell trafficking, such as neuroblastoma 1 (NBL1) and B cell CLL/Lymphoma 6 (BCL6), and cell death, such as Fas cell surface death receptor (FAS) and GTPase IMAP family member 8 (GIMAP8), were differentially expressed in the duodenum of treated and control broilers (P < 0.05). We have demonstrated that C. perfringens culture supernatant (NetB positive) infusion resulted in histological and gene expression changes consistent with necrotic enteritis in the duodenum of broilers. In the absence of live bacteria, crude culture supernatant resulted in early immunomodulation, inflammation, and cell death in the duodenum. The pathways identified here can be targeted for the development of new drugs, vaccines, and novel therapies for necrotic enteritis in broilers.

Designer egg evaluation in a controlled trial.

OBJECTIVE: To evaluate the ability of designer eggs enriched in vitamin E, lutein, selenium (Se) and docosahexaenoic acid (DHA) to deliver micronutrients to the human in a palatable and visually acceptable form. DESIGN: Double-blind, placebo-controlled trial, two treatment groups balanced for sex and age. SETTING: Department of Biochemistry and Nutrition, SAC, Scotland. SUBJECTS: Forty healthy adult volunteers completed the study. Volunteers were recruited among staff of the Scottish Agricultural College Interventions: Volunteers consumed, for 8 weeks, either a designer egg or a normal table egg per day. Fasting blood samples were taken before and at the end of the study. RESULTS: Consumption of designer eggs enriched in vitamin E, lutein, Se and DHA significantly increased the levels of alpha-tocopherol, lutein and DHA in plasma as compared to the changes found after consumption of normal table eggs, with the largest increases found in plasma lutein (1.88-fold increase). The proportion of DHA was increased in all the main lipid classes of the plasma including triacylglycerol (2.3-fold), free fatty acids (1. 6-fold), cholesteryl ester (1.4-fold) and phospholipid (1.3-fold). Egg consumption did not change Se concentration in plasma, blood pressure, total plasma lipid concentrations or the concentrations of total cholesterol and HDL-cholesterol in plasma. CONCLUSION: Consumption of designer eggs enriched in vitamin E, lutein, DHA and Se as part of normal diet for 8 weeks effectively increased the blood levels of alpha-tocopherol, lutein and DHA. SPONSORSHIP: Scottish Office Agriculture, Environment, and Fisheries Department.

The effects of dietary supplementation with docosahexaenoic acid on the phospholipid fatty acid composition of avian spermatozoa.

This study represents an attempt at the dietary manipulation of the fatty acid composition of chicken spermatozoa in order to enhance the levels of n-3 polyunsaturates at the expense of the n-6 fatty acids, which normally predominate in the lipids of avian semen. Male chickens were provided with either a control diet supplemented with maize oil or the test diet supplemented with fish oil (Tuna Orbital Oil) from 10 weeks of age. Semen samples were collected from the birds after 30 and 48 weeks of supplementation. The fish oil diet induced a significant but limited increase in the proportion of 22:6n-3 in the spermatozoan phospholipid in parallel with an equivalent decrease in the proportions of 20:4n-6 and 22:4n-6. However, since the maximal level of 22:6n-3 in the phospholipid that was achieved by fish oil feeding was less than 10% (wt/wt of fatty acids), these changes fell far short of representing a switch from the typical avian pattern to that more characteristic of the n-3 enriched mammalian semen. Analysis of the fatty acid compositions of the constituent classes of phospholipid in the spermatozoa indicated that, in both dietary states, the phosphatidylethanolamine fraction contained much greater proportions of n-6 and n-3 C20-22 polyunsaturates than the phosphatidylcholine fraction. The results indicate that the typical fatty acid profile of the spermatozoa of domesticated poultry, characterised by the predominance of C20-22 n-6 polyunsaturates, displays a considerable degree of resistance to manipulation by dietary means and does not adopt the "mammalian" type of profile following supplementation with n-3 fatty acids.

Fatty acid composition, glutathione peroxidase and superoxide dismutase activity and total antioxidant activity of avian semen.

This work demonstrates that spermatozoa from five avian species (chicken, turkey, guinea fowl, duck and goose) are all characterised by high proportions of polyunsaturated fatty acids, from 46 (turkey) to 55% (duck) of total. For each of the species, the most abundant fatty acids were arachidonic (20:4n-6) and docosatetraenoic (22:4n-6) acids, representing between 22 (turkey) and 40% (chicken) of total. Significant activities of the major isozymes of superoxide dismutase and glutathione peroxidase, which protect against the peroxidation associated with high degree of fatty acid unsaturation, were found in spermatozoa from all species. The seminal plasma also had these activities and showed additional mechanisms for protecting spermatozoa from peroxidation. In general terms, these lipid and enzyme proteins were similar between the five avian species and different from those reported for mammalian sperm.

Carotenoid discrimination by the avian embryo: a lesson from wild birds.

The concentrations (microg/g wet yolk) of total carotenoids in eggs of the common moorhen (Gallinula chloropus), American coot (Fulica americana) and lesser black-backed gull (Larus fuscus), collected in the wild, were 47.5, 131.0 and 71.6, respectively. In contrast to data for eggs of the domestic chicken, beta-carotene was a significant component in the yolks of these three wild species, forming 25-29% by wt. of the total carotenoids present. The concentration of total carotenoids in the livers of the newly-hatched chicks was 5-10 times higher than in the other tissues and beta-carotene was again a major component, forming 37-58% of the hepatic carotenoids. In the newly-hatched gull, the proportions of both lutein and zeaxanthin were very low in the liver but high in the heart and muscle when compared with the yolk. By contrast canthaxanthin, echinenone and beta-carotene were very minor constituents of heart and muscle when compared with their proportions in the yolk of the gull. The proportions of lutein and zeaxanthin in the liver of the newly-hatched coot and moorhen were also far lower than in the yolk whereas the liver was relatively enriched with beta-cryptoxanthin, beta-carotene and (in the moorhen) echinenone. The results indicate that avian embryos discriminate between different carotenoids during their distribution from the yolk to the various tissues.

Phospholipid fatty acid composition, vitamin E content and susceptibility to lipid peroxidation of duck spermatozoa.

Recent studies on chicken semen have suggested that the lipid and fatty acid composition of spermatozoa may be important determinants of fertility. Phospholipid fatty acid composition, vitamin E content and in vitro susceptibility to lipid peroxidation of duck spermatozoa were investigated using GC-MS and HPLC based methods. The total phospholipid fraction of duck spermatozoa was characterized by high proportions of the n-6 polyunsaturated fatty acids arachidonic (20:4n-6), docosatetraenoic (22:4n-6) and docosapentaenoic (22:5n-6) acids but a substantial proportion of the n-3 fatty acid docosahexaenoic (22:6n-3) acid was also present. Palmitic (16:0) and stearic (18:0) fatty acids were the major saturates in sperm phospholipids. Among the phospholipid classes, phosphatidylserine (PS) had the highest degree of unsaturation due to very high proportions of 22:6n-3, 22:5n-6, 22:4n-6 and 20:4n-6, comprising together more than 75% of total fatty acids in this fraction. Phosphatidylethanolamine (PE) also contained high proportions of these four C(20-22) polyunsaturates, which together formed 60% of total fatty acids in this phospholipid. Spermatozoa and seminal plasma of duck semen were characterized by unexpectedly low content of vitamin E, being more than 4-fold lower than in chicken semen. In duck semen the major proportion of the vitamin E (>70%) was located in the spermatozoa. The very high proportion of 22:6n-3 in PS and PE fractions of duck sperm lipids and the comparatively low levels of vitamin E could predispose semen to lipid peroxidation. Nevertheless the in vitro susceptibilities to Fe2+-stimulated lipid peroxidation of duck and chicken spermatozoa were very similar. The results of the study suggest that increased superoxide dismutase and glutathione peroxidase activity and increased antioxidant activity of seminal plasma may compensate for the low levels of vitamin E to help protect the membranes of duck spermatozoa, which exhibit a high degree of unsaturation from oxidative stress.

Tissue-specific antioxidant profiles and susceptibility to lipid peroxidation of the newly hatched chick.

The hatching process is characterized by a range of adaptive changes, and a newly hatched chick is considered as an intermediate stage between prenatal and postnatal development. The aim of the present study was to evaluate the characteristic relationships between tissue-specific fatty acid composition and antioxidant protection in newly hatched chicks. Liver, yolk sac membrane, heart, kidney, lung, and four brain regions (cerebrum, cerebellum, stem, and optic lobes) were collected. Fatty acid composition of total lipids and phosphoglycerides, alpha-tocopherol, lutein, ascorbic acid, reduced glutathione, and the activities of Mn- and Cu,Zn-superoxide dismutase (SOD) and Se-dependent and non-Se-glutathione peroxidase (GSH-Px), and catalase (CAT) were determined. The levels of Fe, Cu, Zn, and Mn as well as tissue susceptibility to lipid peroxidation were also studied. The tissues of the newly hatched chick showed distinctive features in fatty acid profiles, antioxidant accumulation, and susceptibility to lipid peroxidation. The brain clearly displayed the greatest susceptibility to spontaneous and Fe-stimulated lipid peroxidation, was highly unsaturated and contained very low levels of vitamin E, no detectable carotenoids, low GSH-Px, and low CAT activity. At the same time, the brain was characterized by high ascorbic acid concentration and comparatively high SOD activity. It was suggested that in postnatal development, antioxidant enzymes presumably play the major role in antioxidant protection of the chick tissues.