Are baseline and short-term corticosterone stress responses in free-living amphibians repeatable?

Amphibians respond to environmental stressors by secreting corticosterone, a stress hormone which promotes physiological and behavioral responses. Capture handling can be used to stimulate physiological stress response in amphibians. The use of single blood sampling and presentation of mean data often limits the quantification of within and between individual variation in baseline and short-term corticosterone stress responses in amphibians. It is important for studies of amphibian physiological ecology to determine whether baseline and short-term corticosterone stress responses are consistent or not. We quantified repeatability (r), a statistical measure of consistency, in baseline and short-term corticosterone stress responses to a standard capture and handling stress protocol in free-living adult male cane toads (Rhinella marina). Corticosterone metabolite concentrations were measured entirely non-invasively in male toad urine samples via an enzyme-immunoassay. During the first sampling occasion, urine samples were collected manually from individual male toads (n = 20) immediately upon field capture. Toads were handled for 5 min then transferred to plastic bags (constituting a mild stressor), and urine samples were collected hourly over 8 h in the field. The toads were resampled for baseline (0 h) urine corticosterone with hourly urine sampling over 8 h (for quantification of the stress induced corticosterone) at 14 day intervals on three consecutive occasions. Within and between sample variations in urinary corticosterone metabolite concentrations were also quantified. All toads expressed a corticosterone stress response over 8 h to our standard capture and handling stress protocol. Variations both within and between toads was higher for corrected integrated corticosterone concentrations than corticosterone concentrations at baseline, 3 or 6 h. Baseline urinary corticosterone metabolite concentration of the male toads was highly repeatable (r = 0.877) together with high statistical repeatabilities for 3 h (r = 0.695), 6 h (r = 0.428) and 8 h (r = 0.775) corticosterone metabolite concentrations, and for the total and corrected integrated corticosterone responses (r = 0.807; r = 0.743 respectively). This study highlights that baseline and short-term corticosterone stress responses are repeatable in free-living amphibians. Future studies should utilize this non-invasive tool to explore repeatability among seasons and across years, and determine its functional significance in relation to behavioral ecology and reproduction in amphibians generally.     

Experimentally induced stress,oxidative load and changes in immunity in a tropical wild bird,Perdicula asiatica: involvement of melatonin and glucocorticoid receptors

Throughout the year, birds encounter various environmental challenges such as extreme temperatures, rainfall and shortage of food. Here we report on the effect of stress on the general immunity of wild birds as measured by several assays including melatonin, an anti-stress hormone. We selected Perdicula asiatica, a wild tropical bird, and exposed them to experimental stressors such as water deprivation, food deprivation and immobilization, i.e., stressors that they would encounter in a natural environment. We measured the oxidative load in the spleen in terms of superoxide dismutase (SOD), catalase activity and thiobarbituric acid-reactive substance (TBARS) levels. The immune status was judged by total leukocyte count (TLC), heterophil/lymphocyte ratios (H/L) and percent stimulation ratio of splenocytes (%SR). The peripheral levels of melatonin and corticosterone were also determined and correlated with the expression of melatonin (Mel_1a/Mel_1b) and glucocorticoid receptors. Our results showed a significant (p < 0.05) decrease in splenic SOD and catalase activity, while a significant (p < 0.05) increase in TBARS and a corticosterone level was observed. Stressful conditions also decreased the immune status as reflected by the low values of H/L ratios, TLC and %SR. In contrast, melatonin pretreatment significantly (p < 0.05) reduced the oxidative stress and improved the immune parameters when compared to untreated control birds. This suggests that melatonin prevents/alleviates oxidative damage and suppresses the immune status induced by stressful conditions via its membrane receptor expression (Mel_1a and Mel_1b) in P. asiatica.     

Environmental enrichment exerts anxiolytic effects in the Indian field mouse (Mus booduga)

Environmental enrichment (EE) is known to have behavioral and physiological anxiolytic effects in several animal models. However, it is as yet unclear how EE modulates behavior of wild animals and the underlying molecular mechanisms. The adult male field mouse Mus booduga (n = 42) captured at agricultural field, were housed in non-enriched standard condition (SC) for 7 days and considered as directly from wild (DW). Another two groups of mice were housed in either EE or SC for 30 days. Behavioral testing was carried out to assess their anxiety-like behavior in the elevated plus-maze (EPM). We found that on EPM, mice housed in EE display less anxiety like behavior when compared to mice housed in SC. Exposure to plus-maze did not increase the levels of corticosterone (CORT) in prefrontal cortex (PFC) and circulating CORT, and adrenocorticotropic hormone (ACTH) in the mice housed in EE but not in the mice housed in SC. We observed a trend in the EE induced inhibition of expression of corticotropin releasing hormone (CRH), glucocorticoid receptor (GR), E2 ubiquitin-conjugating enzyme (Ubc9) and steroid receptor coactivator-1 (SRC-1) mRNA levels, which are all known to be involved in the stress response signaling pathway. Our study suggests that EE exerts therapeutic and anxiolytic effects against stressors.     

Artificial rain and cold wind act as stressors to captive molting and non-molting European starlings (Sturnus vulgaris)

Free-roaming animals continually cope with changes in their environment. One of the most unpredictable environmental phenomena is weather. Being able to respond to weather appropriately is crucial as it can be a threat to survival. The stress response, consisting of increases in heart rate and release of glucocorticoids, is an important mechanism by which animals cope with stressors. This study examined behavioral, heart rate, and corticosterone responses of captive European starlings (Sturnus vulgaris) to two aspects of weather mimicked under controlled conditions, a subtle (3 °C) decrease in temperature and a short, mild bout of rain. Both decreased temperature and exposure to rain elicited increases in heart rate and corticosterone in non-molting starlings. Molt is an important life history stage in birds that affects feather cover and may require a different response to weather-related stressors. We repeated the experiment in molting starlings and found increases in heart rate in response to rain and cold wind. However, the hypothalamic–pituitary–adrenal (HPA)-axis was suppressed during molt, as molting starlings did not increase corticosterone release in response to either stimulus. These data suggest these stimuli induce increased allostatic load in starlings, and that animals may adjust their response depending on the life-history stage.     

Comparison of the lipid properties of captive,healthy wild,and pansteatitis-affected wild Nile crocodiles (Crocodylus niloticus)

The results presented describe and compare the fatty acid composition and melting properties of captive, healthy wild, and pansteatitis-affected wild crocodiles (Crocodylus niloticus). Differences in fatty acid composition between intramuscular and adipose fat is noted in captive crocodiles, and the latter differs from wild crocodiles as a result of different diets. Adipose fat of healthy wild crocodiles differs minimally from diseased ones, respectively with 37.3 ± 2.6% vs. 43.2 ± 2.3% monounsaturated fatty acids, and 43.2 ± 2.9% in dead crocodiles, while polyunsaturated fatty acids decrease from 27.3 ± 1.9% to as low as 21.9 ± 3.6% respectively. Of the unsaturated fatty acids 18:2n? 6 decreased from 6.5 ± 2.6% in unaffected crocodiles to 3.5 ± 0.6% in highly affected and 3.2 ± 0.4% in dead crocodiles, and 22:5n?3 from 2.8 ± 0.6% to 1.8 ± 0.3% and 2.2 ± 0.3% respectively. The melting properties as determined by differential scanning calorimetry show that extracted adipose fat is a small degree softer in pansteatitis-affected tissue, specifically in the temperature range 7–36 °C, and does not contribute to the hard texture noted for adipose fat tissue of pansteatitis-affected animals. A high moisture content of 51.0 ± 19.7% of the fat tissue of pansteatitis-affected animals vs.17.1 ± 8.0% of healthy ones, suggests that physiological changes due to interstitial inflammation may contribute to the hard texture.     

Behavioural effects of embryonic exposure to corticosterone in chickens

We tested the hypothesis that exposure of chick embryos to corticosterone leads to increased fear, reduced competitive ability, reduced ability to cross a barrier and reduced growth in juvenile chicks. Behaviour was studied in birds subjected to three different egg injection treatments: a negative control (no treatment of eggs), a positive control (100 μl sesame oil vehicle) and a corticosterone treatment (0.6 μg corticosterone in 100 μl sesame oil). Eggs were injected prior to incubation and the behaviour of chicks was studied during the first 4 weeks of life. Corticosterone treatment increased fear in chicks, as indicated by greater avoidance of an observer in the home pen at 2 weeks of age (P < 0.0001), reduced ability to cross a wall to access feed at 2 weeks of age (P < 0.05) and reduced ability to compete for a wormlike object at 4 weeks of age (P < 0.01). Treatment with corticosterone also reduced body weight at 1 week of age (P < 0.003) and 4 weeks of age (P < 0.04), but not at hatch (P < 0.28). The sesame oil vehicle reduced fear (P < 0.0001), but had no other significant effects. These results indicate that embryonic exposure to corticosterone leads to behavioural and growth deficits in chicks.     

Evaluating the safety of Rodolia cardinalis to two species of Galapagos finch; Camarhynchus parvulus and Geospiza fuliginosa

Some species of coccinellids exude a fluid that contains an alkaloid which can be toxic to vertebrates that ingest them. Because of this, before introducing Rodolia cardinalis to the Galapagos Islands as a biological control agent for the invasive scale insect, Icerya purchasi, it was considered necessary to determine whether it is toxic to birds. In this study, experiments were carried out with two common, insectivorous finches that are endemic to the Galapagos Islands; Camarhynchus parvulus (12 individuals) and Geospiza fuliginosa (16 individuals). Finches were maintained in captivity for 12–18 days. This period was divided into three phases: adaptation, experimental, and observation. During the experimental phase, test individuals, in addition to their regular diet, were fed 0.15 g portions of larvae or adults of R. cardinalis on alternate days over four days. The control group were fed 0.15 g of Asciodes quietalis abdomens in place of R. cardinalis. Symptoms of toxicity (mortality, deterioration, critical weight loss) from feeding on R. cardinalis were not observed in either species while in captivity, but birds avoided or rejected beetles indicating that they were distasteful. Finches regurgitated a higher number of larvae than adult beetles suggesting that larvae were less palatable than adults. Although long-term monitoring was not possible, results suggest that at least two species of finch will be unaffected by the release of R. cardinalis into the Galapagos Islands.     

Potentiation of proline accumulation in oilseed rape leaf discs exogenously supplied with combinations of PEG and cryoprotective agents is associated with overproduction of ABA

Physiological processes involved in the control of the magnitude of the stress-induced proline (Pro) response of higher plants are not fully understood. Here we are dealing with Pro accumulation by rape leaf discs (RLDs) treated in vitro, under light conditions, with dual unusual systems containing low concentrations of the non-permeant polymer PEG 6000 (PEG) added with readily permeant cryoprotective agents (CPAs). At osmotically active concentrations, dimethylsulfoxide (DMSO), glycerol, 1,3-propanediol, ethylene glycol and dimethylformamide (DMF) behaved as very poor inducers of the Pro response when provided alone. On the contrary when all those substances (apart DMF) were supplied at onset of treatments, in combination with PEG, Pro levels greatly exceeded what could be predicted through simple additivity of the effects of those substances provided individually. This suggested potentiation effects at the level of some component(s) of Pro metabolism involved in stress-induced Pro accumulation. We have also demonstrated that exogenous ABA could substitute for DMSO (but not PEG), this other binary system also inducing (through potentiation) high rate of Pro accumulation. The strinking similarity between the responses induced with PEG + DMSO and PEG + ABA suggested that DMSO induced increases in the endogenous amount of ABA which might be, at its turn, acting as exogenously supplied ABA. Additional amounts of ABA actually accumulated in leaf discs treated with the dual system PEG + DMSO. Other CPAs tested in this study, except dimethylformamide (DMF), might also be acting on the same way. Potentiating effects, associated with enhanced amount of ABA, were also found to result from combinations of PEG with a range of organic and inorganic substances which mimick to some extent the cytosolute composition of plant cells. Together, our results gain some insight into the physiological mechanisms involved in the control of stress-induced Pro accumulation and strongly suggest their relationship with stress-induced changes in ABA content.     

Corticosterone decreases the activity of rat glutamate transporter type 3 expressed in Xenopus oocytes

Glucocorticoids can increase the extracellular concentrations of glutamate, the major excitatory neurotransmitter. We investigated the effects of corticosterone on the activity of a glutamate transporter, excitatory amino acid carrier 1 (EAAC1; also called excitatory amino acid transporter type 3 [EAAT3]), and the roles of protein kinase C (PKC) and phosphatidylinositol 3-kinase (PI3K) in regulating these effects. Rat EAAC1 was expressed in Xenopus oocytes by injecting mRNA. l-Glutamate (30 μM)-induced membrane currents were measured using the two-electrode voltage clamp technique. Exposure of these oocytes to corticosterone (0.01–1 μM) for 72 h decreased EAAC1 activity in a dose-dependent fashion, and this inhibition was incubation time-dependent. Corticosterone (0.01 μM for 72 h) significantly decreased the V_max, but not the K_m, of EAAC1 for glutamate. Furthermore, pretreatment of oocytes with staurosporine, a PKC inhibitor, significantly decreased EAAC1 activity (1.00 ± 0.06 to 0.70 ± 0.05 μC; P < 0.05). However, no statistical differences were observed between oocytes treated with staurosporine, corticosterone, or corticosterone plus staurosporine. Similar patterns of responses were achieved by chelerythrine or calphostin C, other PKC inhibitors. Phorbol-12-myristate-13-acetate (PMA), a PKC activator, inhibited corticosterone-induced reduction in EAAC1 activity. Pretreating oocytes with wortmannin or LY294002, PI3K inhibitors, also significantly reduced EAAC1 activity, but no difference was observed between oocytes treated with wortmannin, corticosterone, or wortmannin plus corticosterone. The above results suggest that corticosterone exposure reduces EAAC1 activity and this effect is PKC- and PI3K-dependent.     

Application of the index of marsh bird community integrity to coastal wetlands of Georgian Bay and Lake Ontario,Canada

Ecological indicators have gained increasing attention within the scientific community over the past 40 years. Several taxonomic groups have been used successfully as indicators including most prominently fish, invertebrates, plants, and birds because of their ability to indicate environmental changes. In the Laurentian Great Lakes region, there has been recent concern over the applicability of using indicators on a basin-wide scale due to species range restrictions and lake-based differences. The objective of this study was to determine the ability of the Index of Marsh Bird Community Integrity (IMBCI) to indicate land use disturbance surrounding coastal marshes of Georgian Bay and Lake Ontario. To meet this objective, we surveyed birds and vegetation at 14 marshes in Georgian Bay (low land use disturbance) and Lake Ontario (high land use disturbance). Even though Lake Ontario marshes were surrounded by significantly more altered land than Georgian Bay marshes, and had poorer water quality, we found significantly fewer birds in Georgian Bay marshes (mean = 8.2) compared to Lake Ontario (mean = 13.7) and no significant difference in IMBCI scores. This inconsistency could be due to vegetation differences affecting the strength of the index, because Georgian Bay wetlands had significantly more bulrush (Schoenoplectus spp.) and floating vegetation, while Lake Ontario wetland vegetation was taller and cattail-dominated (Typha spp.). These findings suggest that the IMBCI may not be useful on a basin-wide scale in the Great Lakes region in detecting human disturbance surrounding wetlands.     

Corticosterone may interact with peripubertal development to shape adult resistance to social defeat

Studies of social stress in adult mice have revealed two distinct defeat-responsive behavioral phenotypes; “susceptible” and “resistant,” characterized by social avoidance and social interaction, respectively. Typically, these phenotypes are observed at least 1 day after the last defeat in adults, but may extend up to 30 days later. The current study examined the impact of peripubertal social defeat on immediate (1 day) and adult (30 day) social stress phenotypes and neuroendocrine function in male C57BL/6 mice. Initially, peripubertal (P32) mice were resistant to social defeat. When the same mice were tested for social interaction again as adults (P62), two phenotypes emerged; a group of mice were characterized as susceptible evidenced by significantly lower social interaction, whereas the remaining mice exhibited normal social interaction, characteristic of resistance. A repeated analysis of corticosterone revealed that the adult (P62) resistant mice had elevated corticosterone following the social interaction test as juveniles. This was when all mice, regardless of adult phenotype, displayed equivalent levels of social interaction. Peripubertal corticosterone was positively correlated with adult social interaction levels in defeated mice, suggesting early life stress responsiveness impacts adult social behavior. In addition, adult corticotropin-releasing factor (CRF) mRNA in the paraventricular nucleus of the hypothalamus (PVN) was elevated in all defeated mice, but there were no differences in CRF mRNA expression between the phenotypes. Thus, there is a delayed appearance of social stress-responsive phenotypes suggesting that early life stress exposure, combined with the resultant physiological responses, may interact with pubertal development to influence adult social behavior.     

Does sex matter? Differential responses to corticosterone administration in the zebra finch

Because of potentially deleterious effects of chronic stress, physiological measurements of stress hormones (in birds, corticosterone (CORT)) are often used to determine the consequences of natural or human-induced change. Often, it is assumed that CORT levels will be similar between the sexes and the results are pooled. However, recent studies have reported sex differences in CORT concentrations in avian species. As zebra finches (Taeniopygia guttata) are one of the most widely used bird species in laboratory studies worldwide, potential sex-specific differences in hormone metabolism, as well as the clearance rate of oral doses of exogenous CORT, are highly relevant. The results of this study show that female zebra finches have a significantly higher baseline CORT than males, which could partially be a product of differential responses to semi-isolation. In addition, a single dose of exogenous CORT resulted in different blood profiles between the sexes over time, though exogenous CORT was cleared from blood within 90 min following treatment in both sexes. Interestingly, exposure to multiple doses of exogenous CORT resulted in elevated CORT levels 24 h after treatment in both sexes. These results highlight the need for further investigations into potential sex differences in hormone metabolism, as well as possible cumulative effects of repeated stress.     

A novel flexible method to split feeding behaviour into bouts

Before meal patterns can be analysed properly, a biologically relevant meal criterion must be determined in order to group short-term feeding behaviour into meals. Existing methodologies are based on modelling of the frequency distributions of intervals between feeding events but these methods cannot be used if the proper distributions cannot be clearly identified. For such cases we developed two new methods – (1) based on the analysis of the distribution of between-meal interval lengths only and (2) based on the analysis of changes in the probability of animals starting to feed with time since the last feeding event. Both methods were developed using a data set of over 700,000 records of visits to feeders obtained with broilers (Gallus gallus) aged between 2 and 5 weeks. The two methods resulted in meal criteria estimates of 20.1 and 17.5 min, respectively, which, when applied to the data set, gave statistically significant but very small differences in meal characteristics. The new methods were tested against an independent cow (Bos taurus) data set and the resultant meal criteria compared with those predicted by an existing method. The two novel methods estimated meal criteria for cows at 27.9 and 35.5 min, compared with 28.9 min for the existing method. Again, these differences in meal criteria resulted in only very small differences in meal characteristics. Even though meal criteria were relatively similar for birds and cows, characteristics such as average daily number of meals (10.9 and 5.9), meal size (12.5 g and 7.4 kg) and meal duration (7.7 and 31.4 min) were very different. The analyses show that, if the appropriate distributions of intervals cannot be identified, meal criteria can still be estimated for species as diverse as mature cows and young birds by the novel methodologies developed here.     

Behavioral and mortality responses of the bivalves Scrobicularia plana and Cerastoderma edule to temperature,as indicator of climate change's potential impacts

Temperature is one of the most important abiotic factors affected by climate change. It determines physiological processes, ecological patterns and establishes the limits of geographic distribution of species. The induced thermal stress frequently results in physiological and behavioral responses and, in extreme cases, may lead to mortality episodes. Scrobicularia plana and Cerastoderma edule behavioral and mortality responses to temperature were evaluated. Specimens were sampled in the Mondego estuary (Portugal), acclimated and exposed to different temperature treatments (5–35 °C). Individual activity and mortality were registered during 120 h laboratory assays. Both species showed a thermal optimum for their activity (S. plana: 15–23 °C; C. edule: 20–23 °C), and survival was mainly affected by high temperature (S. plana: LC50_120 h = 28.86 °C; C. edule: LC50_120 h = 28.01 °C), with 100% mortality above critical values (≥32 °C). Results further indicated that both species are more affected the higher the temperature and the longer the exposure time. This study indicates that the occurrence of extreme climatic events, especially heat waves, may be particularly impairing for these species.     

Quantifying human disturbance on antipredator behavior and flush initiation distance in yellow-bellied marmots

Human disturbance may differentially affect the behavior of wild animals and such behavioral perturbations may have fitness consequences. To understand the effects of specific types of human disturbance on antipredator behavior, a behavior whose performance enhances survival, we studied yellow-bellied marmots (Marmota flaviventris). We quantified both antipredator vigilance and the flight initiation distance of the marmots to an approaching human in six different colony sites where we also quantified the frequency and type of human visitation. We developed an analysis framework, using linear mixed models, and found that: (1) when the presence of motorized vehicles and bicycles was high, marmots increased the proportion of time spent vigilant (pseudo R² = 0.33 and 0.31 for motorized vehicles and bicycles, P < 0.05) and decreased the time spent foraging (pseudo R² = 0.29 and 0.23 for motorized vehicles and bicycles, P < 0.05), (2) there was no significant effect of the presence of pedestrians on the time allocated to vigilance and foraging (pseudo R² = 0.25 and 0.19, P > 0.05), (3) marmots decreased the flight initiation distance as disturbance of motorized vehicles (pseudo R² = 0.85) and pedestrians (pseudo R² = 0.84) increased (P < 0.05), and (4) when we considered bicycles as the disturbance, juveniles tolerated closer approaches than adults or yearlings (P < 0.001). Marmots thus responded to some human disturbance by adjusting time spent in foraging and shortening the tolerance distance. Since these behavioral responses could have significant implications for survival and reproduction, we should generally view human disturbance as something that can influence natural antipredator behavior. Importantly, based on an understanding of the differential effects of human activities on wildlife, reducing human disturbance should be taken into account for wildlife management. In addition, our approach will be useful to quantify differential effects of humans on wildlife and to enhance our ability to manage those impacts.     

Evidence that metyrapone in the presence of inflammation modulates cytokine mRNA expression

ObjectiveMetyrapone (MT) has been used clinically to decrease glucocorticoid levels in human and animal studies. However, the potential effects of MT in the presence of inflammation are poorly understood. Thus, the aim of this study was to evaluate the effects of the administration of MT on the mRNA levels of pro-inflammatory cytokines in the presence of inflammation induced by the well-established model of ligature-induced periodontitis in rats.Material and methodsSixty animals were randomly assigned into three experimental groups of 20 rats each: G1-control; G2-periodontal disease (PD) induced by cotton ligature; G3-PD associated with 3 daily doses of MT (50 mg/kg/3 × 3 h). After 30 days, all animals were killed by decapitation. Blood samples were taken and the concentrations of corticosterone and catecholamines measured. Marginal tissues around ligated and non-ligated teeth were harvested and gene expression was assessed by quantitative polymerase chain reaction technique (qPCR). Moreover, the area of interradicular bone loss (ABL) was histometrically determined.ResultsData analysis showed that: (i) ligature placement resulted in a significant ABL, as compared to non-ligated sites of G1 group; (ii) mRNA levels of all the pro-inflammatory factors assessed (INF-γ, TNF-α, IL-1β and IL-6) were increased in the PD group (G2) (p < 0.05) when compared to G1; (iii) there were no significant differences in corticosterone and catecholamine plasmatic levels between the three groups; (iv) MT administration, in the presence of inflammation, induces an increased ABL and significantly increased mRNA levels of all pro-inflammatory cytokines analyzed (p < 0.05).ConclusionWithin the limits of this study, it can be concluded that MT in the presence of inflammation may modulate expression of pro-inflammatory cytokines, regardless of its effect on plasma corticosterone levels.     

The effects of cage volume and cage shape on the condition and behaviour of captive European starlings (Sturnus vulgaris)

Cage size is widely recognised as an important determinant of captive animal welfare, but in contrast, cage shape has received far less attention. Husbandry recommendations for flying birds state that cages should be long in shape because this allows greater potential for flight. However, so far no studies have investigated the impact of cage shape on the behaviour or welfare of captive flying birds. We measured the effects of cage size and shape on the condition and behaviour of captive wild-caught European starlings (Sturnus vulgaris) using a 2 × 3 factorial design in which birds were individually housed for 1 week in cages of one of two volumes (either medium at 0.3 m³ or large at 1 m³), and one of three shapes (long with an aspect ratio (i.e. length/height) of 3.43), standard with an aspect ratio of 1.72 or tall with an aspect ratio of 0.86). We found effects of cage size and the interaction of cage size and shape on the behaviour and condition of birds. In interpreting the welfare implications of our results we focused on stereotypic behaviour as measured by incidence of somersaulting and a novel statistic that quantifies sequential dependencies in the birds’ locations within the cage. The lowest measures of stereotypic behaviour were recorded in the large cages and the medium long cage. Cage shape was more important in determining the quantity of stereotypic behaviour in the medium cages than the large cages. Our findings support the recommendation that starlings be housed in long-shaped cages.     

The effect of heat stress on intestinal integrity and Salmonella invasion in broiler birds

The intestinal mucosa works as a barrier to protect the internal environment of the animal from bacteria and bacterial toxins found in the gut lumen. Heat stress may harm this function. Therefore, we designed the current experiment to investigate the effect of heat stress on intestinal integrity, physiological and immunological responses and Salmonella invasion in broiler chickens. At 26 days of age, 72 birds were randomly distributed into 3 treatments, with 8 replicates per treatment and 3 birds per replicate. The three treatments were control treatment; kept at thermoneutral environmental conditions (20 ± 2 °C), chronic heat stress treatment (exposed to 30 ± 2 °C; 24 h/day) and acute heat stress treatment (exposed to 35 ±2 °C from 09:00 to 13:00 and kept at 20 ± 1 °C from 13:00 to 09:00). The heat stress exposure was conducted for 10 successive days. Compared with the control treatment, birds subject to chronic and acute heat stress had reduced (P < 0.05) body weight and body gain and increased (P < 0.05) feed conversion ratio. However, feed intake and mortality rate were only increased (P < 0.05) in the acute heat stress treatment. Rectal temperature and Δ rectal temperature (°C/h) increased (P < 0.05) sharply during the first 2 days of exposure followed by gradual decreases until a plateau was achieved. Heat-stressed birds had increased (P < 0.05) serum concentrations of corticosterone, endotoxin lipopolysaccharide and the systemic inflammatory cytokine: TNF-α and IL-2, as well as a higher (P < 0.05) prevalence of Salmonella spp. in meat and livers, as compared with control treatment. It can be concluded that heat stress impaired intestinal integrity which resulted in increased intestinal permeability to endotoxin, translocation of intestinal pathogens (Salmonella spp.) and serum inflammatory cytokines. Therefore, avoiding thermal dysfunction of intestinal barrier is a significant factor in maintaining welfare, immune status and meat safety of broiler birds.     

Genetic characterization of game bird island populations: The conservation of the black francolin (Francolinus francolinus) of Cyprus

The release of exotic genotypes into the wild can lead to the extinction of local demes through the hybridization among distinct gene pools. This may occur between wild specimens and their domesticated relatives. Escaped pets represent a well-known pathway for the introduction of allochtonous resources, and island environments require major attention as they include small-sized and naïve populations that may be prone to extinction. We characterized the genetic make-up of the black francolin (Francolinus francolinus, Phasianidae) resident to Cyprus using microsatellite and mitochondrial DNA for the purpose of sustainable use and long-term protection of this game species. We collected 59 samples of specimens hunted in the districts of Nicosia and Paphos, while others (n = 18) were obtained from birds of unknown origin. These had been imported to a pet-centre in Nicosia, the largest in the capital offering animals either for production or leisure purposes. Both genetic systems pointed to the occurrence of distinct wild district populations that could be treated as separate management units. The non-significant value of the inbreeding coefficient (F_IS) computed for Nicosia and Paphos populations (−0.039 and −0.189, respectively: P > 0.05) and the lack of evidence for recent genetic bottlenecks did not indicate a serious risk of over-hunting. The microsatellites revealed the birds in the pet-centre as highly genetically divergent with respect to the wild ones, the mtDNA pointed to their assignment to alien Asian subspecies once they were compared to141 sequences of allopatric francolins. We advised the Game & Fauna Service to consider law enforcement to ban the import, detention and sale of these subspecies to protect the black francolin population native to Cyprus. The results are discussed in the context of a bibliographic frame including the partridges of the genus Alectoris (Phasianidae) as a model group with reference to genetic characterization and conservation of game bird island populations.     

Allometric scaling of fatty acyl chains in fowl liver,lung and kidney,but not in brain phospholipids

The phospholipid (PL) fatty acyl chain (FA) composition (mol%) was determined in the kidney, liver, lung and brain of 8 avian species ranging in body mass from 150 g (Japanese quail, Coturnix coturnix japonica) to 19 kg (turkey, Meleagris gallopavo). In all organs except the brain, docosahexaenoic acid (C22:6 n3, DHA) was found to show a negative allometric scaling (allometric exponent: B = ? 0.18; ? 0.20 and ? 0.24, for kidney, liver and lung, respectively). With minor inter-organ differences, smaller birds had more n3 FAs and longer FA chains in the renal, hepatic and pulmonary PLs. Comparing our results with literature data on avian skeletal muscle, liver mitochondria and kidney microsomes and divergent mammalian tissues, the present findings in the kidney, liver and lung PLs seem to be a part of a general relationship termed “membranes as metabolic pacemakers”. Marked negative allometric scaling was found furthermore for the tissue malondialdehyde concentrations in all organs except the brain (B = ? 0.17; ? 0.13 and ? 0.05, respectively). In the liver and kidney a strong correlation was found between the tissue MDA and DHA levels, expressing the role of DHA in shaping the allometric properties of membrane lipids.