Immediate and transgenerational effects of thymol supplementation, inactivated Salmonella and chronic heat stress on representative immune variables of Japanese quail.

Environmental challenges are integrated in the inmunoneuroendocrine interplay, impacting the immune system of the challenged individuals, and potentially implying transgenerational effects on their offspring. This study addressed whether dietary supplementation with thymol can modulate the immune response of adult Japanese quail when simultaneously exposed to an inoculum of inactivated Salmonella Enteritidis and a chronic heat stress (CHS). We also evaluated whether the experienced situations by adults can affect the immune response of their undisturbed offspring. In the parental generation, supplemented quail exposed to CHS had a higher inflammatory response and similar values of the heterophil/lymphocyte (H/L) ratio than those that were not supplemented. In their offspring, those chicks whose parents were exposed to CHS showed higher inflammatory response and lower antibody production. Regarding the H/L ratio, chicks whose parents were supplemented showed lower H/L ratio values. Dietary supplementation with thymol partially and positively modulated the inflammatory response and avoided H/L ratio alteration in the parental generation exposed to high environmental temperatures, suggesting these adults were better at dealing with the challenge. The lower H/L ratio values in the offspring suggests that chicks are more capable to deal with potential stressful situations associated with conventional breeding conditions.

Thymol supplementation effects on adrenocortical, immune and biochemical variables recovery in Japanese quail after exposure to chronic heat stress.

Chronic heat stress (CHS) exposure negatively impairs avian' immunoneuroendocrine interplay. Thymol has shown several bioactive properties including antioxidant, bactericidal, antifungal and gamma-aminobutyric acid modulator activities. Indeed, supplementation with thymol has been used with positive effects on poultry production and immune-related variables. This study evaluates whether a thymol dietary supplementation can be used as a new functional feed strategy to mitigate CHS deleterious effects on endocrine, biochemical and immune-related variables. Starting at 100 days of age, 24 fully adult Japanese quail were fed with a diet supplemented with thymol (≈80 mg/quail per day) and other 24 quail remained non-supplemented (control diet). Between 119 and 127 days of age, half of the quail within those groups were submitted to a CHS by increasing environmental temperature from 24°C to 34°C during the light phase and the other half remained at 24°C (non-stressed controls). A period of 3 days after CHS ended (during the recovery period), corticosterone, albumin, total proteins and globulins and glucose concentrations, inflammatory response, antibody production and heterophil to lymphocyte (H/L) ratio were assessed. No differences between groups were found in basal corticosterone concentrations. Total proteins, total globulins and glucose concentrations were found elevated in the previously CHS group compared with their control counterparts. Regardless of the previous CHS exposure, thymol supplementation increased albumin concentrations and inflammatory responses and decreased antibody titers. An interaction between thymol supplementation and prior CHS exposure was found on the H/L ratio. Quail previously exposed to CHS and supplemented with thymol showed similar H/L values than their control non-stressed counterparts, suggesting that thymol has a stress preventive effect on this variable. The present findings together with the already reported thymol bioactive properties, suggest that feed supplementation with this compound could be a useful strategy to help overcoming some of the CHS induced alterations.

Adrenocortical responses in Japanese quail classified by their permanence in proximity to either low or high density of conspecifics.

The density-related permanence test (DRP) is used to classify young birds (while in groups) according to their individual permanence in proximity to either a high or low density of conspecifics (HD or LD, respectively). The birds' performance in DRP was associated with underlying differences in the social responses of their individuals. Quails in homogeneous groups of LD residents responded with less compact groups and higher levels of agonistic interactions to the presence of an intruder, and showed higher levels of agonistic interactions among cagemates than the homogeneous groups of HD birds. This study deepen the characterization of DRP evaluating whether contrasting behavior during test is associated with birds' stress responses to challenging situations during rearing. Plasma corticosterone responses were individually assessed after submitting DRP categorized birds to a brief (10 min) partial restraint (mechanical stressor; experiment 1), or after introducing them during 5 min as intruders in homogeneous groups of 6 unknown same category (HD or LD) conspecifics (social stressor; experiment 2). Experiment 1 showed that basal corticosterone levels were similar in all groups and the partial mechanical restraint induced an increase in plasma corticosterone concentration also in all groups. However, the increase induced by the stressor was higher in the LD birds than in their HD counterparts. In experiment 2, compared to controls, social stressor showed no plasma corticosterone changes in the intruders that were introduced in an unfamiliar group of HD conspecifics. However, intruders (both HD and LD) that were visiting the LD residents showed an increased corticosterone response compared to their control counterparts. Results suggest that categorization of birds in the DRP test could have relevance for selection programs oriented to obtain birds better suited to intensive rearing conditions that includes high density of animals and exposure to unavoidable stressors.

Manipulation of the phenotypic appearance of individuals in groups of laying hens: effects on stress and immune-related variables.

This study evaluated whether phenotypic appearance (PA) alteration during two developmental phases in laying hens, reared in two different group sizes, affects stress and immune responses. After hatching, 750 chicks were randomly assigned to 30 pens at a group size of either 10 or 40 birds. Then, the appearance of 0, 30, 50, 70 or 100% of the chicks in each pen was altered by blackdyeing their head feathers (marked); remaining chicks were unmarked. At 32 weeks, basal and postacute stress plasma corticosterone concentration, leukocyte counts, phytohemagglutinin-p lymphoproliferative and primary antibody responses were measured in six birds/pen. Analysis of variances (ANOVAs) showed no differences among treatment combinations. In a second phase, birds within initially homogeneous pens were sequentially either marked or had dye bleached to alter PA of 70% of hens in each flock (= group in a pen). Hens within initially heterogeneous pens remained unaltered as controls. The above variables were remeasured. Hens in phenotypically manipulated pens showed modified leukocyte counts compared to hens in control pens, indicating a chronic stress reaction in all penmates (whether individual PA was altered or not). Social isolation increased plasma corticosterone concentration. However, within groups of n = 40, phenotypically unaltered hens had lower responses than their altered penmate counterparts, suggesting that remaining in a stable PA group aids better coping with challenges. Although all hens in manipulated pens showed modified leukocyte counts, their antibody and lymphoproliferative responses did not differ from controls suggesting that all groupmates were able to immunologically cope with the challenges presented, within the timeframe evaluated.

Combined effects of aflatoxin B1 and corticosterone treatment on selected performance indices and liver histopathology in Japanese quail.

Animal feed may be contaminated with different mycotoxins, with aflatoxin B(1) (AFB(1)) being a very common and toxic compound. Considering that birds normally have to cope with different stressful situations at the same time, the present study aims to evaluate the effects of feed contamination with AFB(1) in combination with corticosterone treatment in drinking water (a model to induce physiological stress in birds) on selected performance indices: BW, feed conversion, egg production, and macroscopic and microscopic liver alterations. At 5 wk of age, quails were randomly assigned to 1 of 6 dietary treatment groups that resulted from the combination of the presence or absence of corticosterone in drinking water (5 mg/L) with the presence or absence of AFB(1) contamination (0, 100, or 500 µg/kg). The animals remained in these treatments from 5 to 11 wk of age. There were 6 replicates per treatment, each containing 2 males and 2 females. Contamination with 100 µg of AFB(1) per kilogram of feed induced no changes in BW, feed conversion, and egg production parameters. Quail fed with 500 µg of AFB(1) per kilogram of feed showed significant decreases in BW and feed consumption compared with their control counterparts. Corticosterone in combination with 500 µg of AFB(1) per kilogram of feed intensified the negative effects observed on BW and feed consumption and also had negative effects on feed conversion rate and egg production parameters, suggesting that the adverse effects of contamination with AFB(1) are intensified in situations of chronic stress. Quail treated with 500 µg of AFB(1) per kilogram showed hepatocytes with degree 1 and 2 lesions, and all quail treated with 500 µg of AFB(1) per kilogram of feed in combination with corticosterone showed degree 2 liver lesions (i.e., hepatocytes with fatty macro and microvacuoles and necrosis). This result is also consistent with the hypothesis that chronic stress exacerbates the effect of AFB(1) contamination. In conclusion, this study suggests that the negative effects of AFB(1) contamination are increased when overlapped with chronic stressful stimulation.

Effect of feed contamination with aflatoxin B1 and administration of exogenous corticosterone on Japanese quail biochemical and immunological parameters.

Stress is the loss of homeostasis by external forces or stressors. Manipulation, transport, contamination, and other procedures involved in production could be considered stressors. Contamination is a problem commonly faced by producers in the poultry industry. Aflatoxicosis is one of the most common infections resulting from feed contaminated with Aspergillus flavus and Aspergillus parasiticus. This study evaluated the potential effects of the combined administration of aflatoxin B(1) (AFB(1)) and corticosterone on biochemical (concentration of globulins, proteins, and albumin) and immunological (inflammatory response and heterophil:lymphocyte ratio) parameters of Japanese quail. Potential sex effects on those parameters were also considered. The provision of corticosterone in drinking water is a method used for mimicking the effects of chronic stress in avian species. At 35 d of age, 24 mixed-sex groups of 4 animals (2 males and 2 females) were housed in cages and assigned to 1 of 4 treatments: plain drinking water and laying diet, corticosterone administration in drinking water, feed contamination with AFB(1) (100 µg/kg of feed), or corticosterone plus AFB(1) administration. There were 6 cages per treatment. No significant effect of sex in any of the parameters analyzed was detected. Hypoproteinemia, hypoalbuminemia, and hypoglobulinemia were observed in animals treated with corticosterone or contaminated feed. These responses were exacerbated when the factors were combined. The immunodepressive effect of corticosterone administration was confirmed, and a higher effect was noticed when combined with the aflatoxin contamination. Aflatoxin contamination affected birds' physiology similar to a chronic stressor stimulation because it elevates the heterophil:lymphocyte ratio. This study suggests that the effects of the AFB(1) contamination are further increased when overlapped with a chronic stressful stimulation and emphasizes the importance of controlling potential stressor combinations during animal rearing to preserve not only the animal's health status but also their welfare.

Chronic stress and environmental enrichment as opposite factors affecting the immune response in Japanese quail (Coturnix coturnix japonica).

Procedures in the commercial production of animals involve stressful situations which lessen the animal's welfare. This study on Japanese quail evaluated whether an environmental enrichment manipulation can affect avian immune responses and if combined with a chronic stressor exposure can help to counteract the negative effects of stress on the immune system. Potential gender effects were also considered. After hatch, half of the birds were housed in non-enriched boxes and half were housed in environmentally enriched boxes. From day 33 to 42 of age, all birds within half of the non-enriched and enriched boxes remained undisturbed while the other half were daily exposed to a 15 min restraint stressor (chronic stressor). The inflammatory response (lymphoproliferation after phytohemagglutinin-p), percentage of lymphocytes, heterophil/lymphocyte (H/L) ratio and primary antibody response against sheep red blood cells were assessed. The chronic stressor application and the enrichment procedure, respectively, either increased or reduced the four immunological parameters evaluated and always in opposite directions. Males consistently showed lower antibody titres than females and presented the highest H/L ratio in response to the stressor when reared in the non-enriched environment. The findings indicate that submitting these animals to an enriched environment can be effectively used to improve their immune response and to reduce the detrimental effects of a stressor exposure.

Ontogeny of copulatory behaviour in male Japanese quail classified by their T-maze performance as hatchlings.

1. This study examines the time courses of male mating behaviours using quail that were categorised at 2 d of age as high performance (HP) or low performance (LP) individuals in a T-maze. 2. Individually caged males from each T-maze category were observed for 5 min after a female of the same category (HP or LP) was introduced into their home cages as a sexual partner at 4, 5, 6, 8, 10 and 12 weeks of age. The number of grabs, mounts and cloacal contacts performed by each male and his copulatory efficiency (number of cloacal contacts/number of grabs; CE) were determined at each time interval. 3. No differences were observed in the proportion of HP and LP males performing grabs at any of the ages evaluated. However, a greater proportion of HP males showed mounting and cloacal contact behaviour, coupled with a higher CE than did their LP counterparts at 5 and 6, 6 and 8, and 8 weeks of age, respectively. At those ages, a higher mean number of mounts and cloacal contacts were also observed. No significant sexual behaviour differences between HP and LP quail were observed at 10 and 12 weeks of age. 4. The results suggest that rapid negotiation of the T-maze in hatchlings is associated with accelerated puberty in male quail. The differential reproductive behaviour of HP males during the development of puberty does not remain extant in fully developed quail. 5. The finding of accelerated male developmental sexual activity responses in HP quail indicates that selection for T-maze HP behaviour may allow males to be successfully bred at an earlier age.