Ecology: global warming and amphibian losses.

Is global warming contributing to amphibian declines and extinctions by promoting outbreaks of the chytrid fungus Batrachochytrium dendrobatidis? Analysing patterns from the American tropics, Pounds et al. envisage a process in which a single warm year triggers die-offs in a particular area (for instance, 1987 in the case of Monteverde, Costa Rica). However, we show here that populations of two frog species in the Australian tropics experienced increasing developmental instability, which is evidence of stress, at least two years before they showed chytrid-related declines. Because the working model of Pounds et al. is incomplete, their test of the climate-linked epidemic hypothesis could be inconclusive.

Stress reduction through consolation in chimpanzees.

Consolation, i.e., postconflict affiliative interaction directed from a third party to the recipient of aggression, is assumed to have a stress-alleviating function. This function, however, has never been demonstrated. This study shows that consolation in chimpanzees reduces behavioral measures of stress in recipients of aggression. Furthermore, consolation was more likely to occur in the absence of reconciliation, i.e., postconflict affiliative interaction between former opponents. Consolation therefore may act as an alternative to reconciliation when the latter does not occur. In the debate about empathy in great apes, evidence for the stress-alleviating function of consolation in chimpanzees provides support for the argument that consolation could be critical behavior. Consistent with the argument that relationship quality affects their empathic responses, we found that consolation was more likely between individuals with more valuable relationships. Chimpanzees may thus respond to distressed valuable partners by consoling them, thereby reducing their stress levels, especially in the absence of reconciliation.

Laboratory animals. Researchers pained by effort to define distress precisely.

The U.S. Department of Agriculture requested comments earlier this year to help it decide whether to adopt a formal definition of "distress" as part of its responsibilities under the Animal Welfare Act. But the 2600 pieces of advice it received during a 4-month period that ended earlier this month suggest that its job won't be easy. The comments highlight a deep split between animal activists, who see the potential new regulations as a step toward eliminating all painful procedures, and most researchers, who say that the present system is working well and that no major changes are needed.

H blood types in pigs as predictors of stress susceptibility.

At least two genotypes in the H system of blood groups in pigs are responsible for blood types associated with the porcine stress syndrome (PSS), and at least three genotypes are responsible for blood types associated with freedom from PSS. Two blood types, each of which apparently may result from more than one genotype, are associated with PSS in some pigs and not in others.

Cortisol and behavioral responses of working dogs to environmental challenges.

This paper's primary objective is to analyse the physiological (cortisol) and behavioral responses of military working dogs (MWD). Dogs (N=27) were submitted twice to environmental challenges (challenge 1 and 2, 20 days in-between) composed of social (training), visual (mobile toy car) and auditory (air blast) stimuli. Cortisol levels decreased back to the baseline after the second challenge. The behavioral observations showed that these MWD were more active, and presented less stereotypic behaviors (pacing, manipulation of the environment) during both visual challenges, whereas half low posture was observed during the first but not during the second visual challenge. The present study shows that this group of MWD still has an adaptation capacity to an environmental challenge (return to baseline of the cortisol levels, a higher posture during the second than at the first challenge). These results are encouraging and indicate that the dogs might have a diminished welfare (i.e. stereotypic behaviors), but are not chronically stressed.

The possibility of positive selection for both F18(+)Escherichia coli and stress resistant pigs opens new perspectives for pig breeding.

F18(+)Escherichia coli infections causing post-weaning diarrhoea and/or oedema disease are a major cause of economic losses in pig industry. To date, no preventive strategy can protect pigs from F18(+)E. coli infections. One of the most attractive approaches to eliminate F18(+)E. coli infections is the selection for pigs that are resistant to F18(+)E. coli infections. However, this strategy was not believed to be favourable because of reports of genetic association with the stress-susceptibility gene in the Swiss Landrace. To investigate this potential association more thoroughly, 131 randomly selected Belgian hybrid pigs were genotyped for both the F18(+)E. coli resistance alleles (FUT1(A)) and the stress-susceptibility alleles (RYR1(T)) and their association was investigated by determining the linkage disequilibrium. This linkage disequilibrium (LD=-0.0149) is close to zero and does not differ significantly from 0 (likelihood ratio test chi(1)(2)=1.123, P=0.29), demonstrating no association between the FUT1(A) and RYR1(T) alleles. Furthermore, only a small fraction (4.6%) of the Belgian pigs was found to be resistant to F18(+)E. coli infections. Our results suggest that selection for F18(+)E. coli resistant pigs might be an attractive approach to prevent pigs from F18(+)E. coli infections, unlike to what has previously been postulated.

Development of a method of measuring cellular stress in cattle and sheep.

In the current studies, flow cytometric methods were used to demonstrate that heat shock protein (hsp) 70 is constitutively expressed in ovine and bovine leukocytes but that the level of expression varies considerably between different leukocyte types and between species. We also show that expression of hsp70 is upregulated in response to an in vitro heat shock treatment. The optimal temperature for heat shock of leukocytes from sheep and cattle is 43.5 degrees C. In sheep and cattle, the relative susceptibility of leukocyte type to upregulation of hsp70 expression, as assayed as percent positive cells, by in vitro heat shock was cell type specific. Best results were obtained from fresh samples; after storage at room temperature for 24h upregulation was highly variable between animals and less than in fresh samples. These studies demonstrate that evaluation of leukocyte hsp70 expression by flow cytometry is a robust, reproducible method for use in the evaluation of cellular stress responses in cattle and sheep. The application of the methods described may be a valuable tool in assessing in vivo stress responses in livestock species.

Transportation stress alters the circulating steroid environment and neutrophil gene expression in beef bulls.

Stress and its association with altered immune function and incidence of respiratory diseases in cattle have lead to concerns over animal health and welfare during truck transportation. Previously, bulls subjected to transportation stress displayed altered expression of candidate neutrophil genes, warranting a broader investigation of the neutrophil transcriptome and possible associations with fluctuations in circulating steroid hormones. In the current study, blood was collected from six Belgian BluexFriesian bulls at -24, 0, 4.5, 9.75, 14.25, 24, and 48h relative to initiation of 9h of truck transportation. Plasma concentrations of cortisol, dehydroepiandrosterone (DHEA), progesterone, and testosterone were measured; cortisol:DHEA ratios were computed. Neutrophil gene expression was monitored by microarray analysis using bovine immunobiology (BOTL-5) microarrays. Eighty-eight genes were identified as being differentially expressed at P<0.05. Putatively affected genes were grouped into ontological clusters; those of greatest interest for qRT-PCR validation were involved in immune response, apoptosis, wound healing, and several of currently unknown function. Confirmed gene expression changes supported the dramatic effects of transportation stress on the bovine neutrophil transcriptome. Temporal correlations between gene expression profiles and circulating total leukocyte and neutrophil counts were apparent. However, few relationships between gene expression and plasma steroid profiles were detected, possibly due to the biological time-lag between these variables not captured by the blood collection schedule. Further investigation into the factors underlying neutrophil gene expression changes and validations at the protein and cell behavior levels will lead to a better understanding of altered innate immunity in cattle during transportation stress.

Stress alters the cellular and proteomic compartments of bovine bronchoalveolar lavage fluid.

The stresses of transportation, weaning and commingling are associated with an increased incidence of bacterial and viral pneumonia in cattle. Proteins expressed in the epithelial lining fluid (ELF) of the lungs, in conjunction with resident leukocytes, represent the first line of defence against opportunistic pathogens, and stress-induced alterations in their expression may reveal markers of disease susceptibility. Bronchoalveolar lavage fluid was sampled in weaned and transported calves and ELF protein expression was compared to a control group of calves using two-dimensional electrophoresis (2DE). Serum and pulmonary haptoglobin were increased following stress concurrent with the number of blood neutrophils. Using 2DE, significant changes in expression were observed in spots identified by mass spectrometry as annexin A1 and A5, odorant-binding protein (OBP), isocitrate dehydrogenase, fibrinogen, heme-binding protein, alpha-2-HS-glycoprotein, alpha-1-antichymotrypsin and albumin. Quantification of OBP mRNA by real-time RT-PCR and OBP protein by western blot revealed gender-dependent differences in relative OBP expression in response to stress. These findings reveal stress-associated protein changes in pulmonary ELF and suggest a mechanism through which stress alters respiratory disease susceptibility.

Trypanosoma cruzi: effects of social stress in Calomys callosus a natural reservoir of infection.

Social environment can represent a major source of stress affecting cortisol and/or corticosterone levels, thereby altering the immune response. We have investigated the effects of social isolation on the development of Trypanosoma cruzi infection in female Calomys callosus, a natural reservoir of this protozoan parasite. Animals were divided in groups of five animals each. The animals of one group were kept together in a single cage. In a second group, four females were kept together in a cage with one male. In the final group, five individuals were kept isolated in private cages. The isolated animals showed body weight reduction, decreased numbers of peritoneal macrophages, lower global leucocytes counts, smaller lytic antibody percentage and a significantly higher level of blood parasites compared to the other animals. Their behavior was also altered. They were more aggressive than grouped females, or females exposed to the presence of a male. These results suggest that isolation creates a distinct social behavior in which immunity is impaired and pathogenesis is enhanced.

Physiological and behavioral effects of social introduction on adult male rhesus macaques.

Pair housing of laboratory macaques is widely considered to lead to positive changes in well-being, yet the process of introduction is viewed as potentially stressful and risk-prone. Behavioral and physiological data were collected on eight adult male rhesus macaques before, during, and after the process of introduction, in order to measure the initial stress of introduction as well as long-term changes in well-being. Socially experienced subjects, all implanted with biotelemetry devices, were studied in five successive phases: baseline (singly housed), 1 day each of protected contact and full contact introduction, post-introduction (1-3 weeks after introduction), and settled pairs (> or =20 weeks after introduction). One hundred and seventy-six hours of behavioral data and 672 hr of heart rate data were analyzed. Fecal cortisol was also measured for the baseline, post-introduction, and settled pair phases. All introductions were successful and subjects showed no physiological or behavioral signs of stress, such as increased heart rate, abnormal behavior, or psychological indices of distress (depressive/anxiety-related behavior). Agonism was minimal throughout the introduction process and over the subsequent months; only one wound was incurred over the course of the study. Levels of abnormal behaviors, psychological indices of distress, locomotion, inactivity, and affiliation showed improvements within several weeks after introduction; these changes were still present 5-9 months later for the latter two categories. Heart rates during introduction fell significantly in the settled pair phase, and also varied predictably with time of day. Fecal cortisol levels were lower in settled pairs than in single housing. The fact that reductions in abnormal behavior did not persist over the long term may have been confounded by increasing duration of time spent caged. The results of this study may be of practical use for designing and monitoring social introductions and suggest that managers should not dismiss the feasibility of successful pairing of adult male rhesus macaques.

Rams with poor feed efficiency are highly responsive to an exogenous adrenocorticotropin hormone (ACTH) challenge.

An animal's response to a stressor is to increase metabolic rate, and thus energy consumption through the activation of the hypothalamic-pituitary-adrenal axis. Changes to energy use by an animal are likely to influence the efficiency with which it is utilised. In this study, we tested the hypothesis that less efficient sheep are more responsive to exogenous administration of adrenocorticotropin hormone. This was done by firstly determining the appropriate dose (0.4, 1.6 or 6.4microg/kg LW) and peak serum cortisol response time (45min) to exogenous administration of adrenocorticotropin hormone in a pilot study (n=3 sheep). Following this, adrenocorticotropin hormone (2.0microg/kg LW) stimulated cortisol levels were measured in a larger group of sheep (n=50) of known feed efficiency (feed conversion ratio and residual feed intake values). Less efficient sheep (more positive residual feed intake values) were found to have a greater (P<0.001) increase in cortisol concentration in comparison to more efficient animals. Those sheep which had higher levels of cortisol also had a greater proportion (P<0.001) of fat tissue. These data clearly demonstrated that efficiency of energy use, when measured as residual feed intake, is significantly related to an animal's stress response. These findings have important implications for understanding the physiological mechanisms underpinning efficiency of energy use, and may be useful in successfully identifying animals which are superior in terms of feed efficiency.

Stress and demographic decline: a potential effect mediated by impairment of reproduction and immune function in cyclic vole populations.

The stress response is initially adaptive, operating to maintain homeostasis. However, chronic long-term exposure to stressors may have detrimental effects. We proposed that chronic stress may be a major factor in demographic vole cycles, inducing decline in high-density populations. We monitored four populations of the fossorial water vole Arvicola scherman, which undergo pluriannual demographic cycles in the Jura Mountains (France). Sampling was conducted during the high densities and the decline. We measured fecal corticosterone metabolites (FCMs) to assess stress levels and injected phytohemagglutinin to estimate the cell-mediated immune response. We demonstrated that stress levels increase between the high densities and the decline in most of the vole populations. At the individual level, FCM concentrations varied with reproductive status and body condition. During the outbreak, we observed significantly higher levels of FCM concentrations in nulliparous females than in females that had previously reproduced. Moreover, a significant negative correlation was observed between concentrations of FCMs and both immunocompetence and body condition during population decline. These results might reflect an impairment of the female reproductive capability in high densities and accelerated senescence affecting immune function during decline, both arising from chronic stress.

Differential gene expression analysis in European eels (Anguilla anguilla, L. 1758) naturally infected by macroparasites.

We analyzed the relationships between the macroparasite community of the European eel and the expression of genes involved in the host physiology during its continental life. The genes studied are implicated in (1) host response to environmental stress, i.e., heat shock protein 70 (HSP70) and metallothionein (MT); (2) osmoregulation, i.e., beta thyroid hormone receptor (betaTHR) and Na+/K+ATPase; and (3) silvering, i.e., betaTHR, freshwater rod opsin (FWO), and deep-sea rod opsin (DSO). All were enumerated by quantitative reverse-transcription polymerase chain reaction. The epizootiological results for 93 yellow eels caught in the Salses-Leucate Lagoon (France) included 11 species: 1 nematode, 2 acanthocephalans, 1 monogenean, and 7 digeneans. The molecular results revealed (1) a significant negative relationship between digenean abundance and the expression level of all the tested genes, except FWO; (2) a significant negative relationship between the abundance of the nematode Anguillicola crassus and the expression level of the Na+/K+ATPase gene; and (3) a significant positive relationship between the A. crassus abundance and the expression level of the MT gene. Eels infected with digeneans had, on average, a lower level of expressed genes. We hypothesize that the parasites may disturb the eel's ability to withstand environmental stress and delay their migration to the Sargasso Sea because of degeneration of the gut. We further propose that the effect of the invasive species, A. crassus, on the gene expression was mainly linked to an increased trophic activity of infected eels. Moreover, it is possible that the parasite may have an effect on the fish's migratory behavior, which is tied to reproductive purposes. Additional work, including an experimental approach, is required to confirm our hypotheses.

Dexamethasone depletes gammadelta T cells and alters the activation state and responsiveness of bovine peripheral blood lymphocyte subpopulations.

Administration of dexamethasone (DEX) to cattle is commonly used in models of stress-induced effects on host defense, including models investigating interactions of microorganisms with their host. Much less is known about the effects of DEX on the adaptive immune response in cattle compared with other species. The objective of the present study was to characterize subsets of circulating lymphocytes in calves before and 48 h after the onset of parenteral DEX treatment. Treatment significantly reduced the overall percentage of circulating lymphocytes and disproportionately depleted the population of gammadeltaTCR(+)/CD8alpha(-) cells. Analysis within the CD8alpha(+) population of T cells revealed that DEX treatment also reduced the CD8alpha(low) subset of gammadeltaT cells coexpressing the activation marker ACT-2(+). By contrast, DEX treatment did not affect the percentage of CD8alpha(low)/CD25(+) cells, indicating that cells with a special activation state were affected. Dexamethasone treatment reduced the number of gammadeltaT cells but increased the percentages of CD14(+) monocytes and activated CD25(+) cells (both CD4(-) and CD4(+)) in peripheral blood mononuclear cell (PBMC) preparations. Although DEX treatment reduced the overall proliferative capacity of PBMC, it enhanced the relative number of proliferating CD4(+) lymphocytes. Lower levels of mRNA for several Th-prototype cytokines (IL-2, IFN-beta, IL-4, transforming growth factor-beta) were detected in short-term PBMC cultures established from DEX-treated calves compared with PBMC cultures from control calves; the amount of il-10 transcripts, however, was unaffected. Results of the study reported here clearly show that DEX treatment does not uniformly suppress the bovine immune system but has differential effects on lymphocyte sub-populations and functions. This information must be considered when using DEX treatment as a bovine stress model.

Effects of road transportation on lymphocyte subsets in calves.

The effect of transportation on peripheral blood lymphocyte subsets in 24 calves was investigated by flow cytometry. Blood was collected before departure, on arrival, at 24h and 1 week after arrival. Highest leucocyte and neutrophil counts, associated with increased concentrations of cortisol and catecholamines, indicated that stress was maximal upon arrival. At this time, a decrease in the percentages of all T lymphocyte subsets was evident, while they did not decrease as absolute counts. The proportion of CD21(+) cells did not change, indicating that the relative reduction of T lymphocyte subsets was not related to an increase in B lymphocytes. These variations may be due to the increase of a natural killer (NK) cell subset. NK cell expansion, together with increasing lymphocyte count and increasing major histocompatibility complex class II expression, may indicate stress-induced stimulation of the immune system.

Climate change impacts and risks for animal health in Asia.

The threat of climate change and global warming is now recognised worldwide and some alarming manifestations of change have occurred. The Asian continent, because of its size and diversity, may be affected significantly by the consequences of climate change, and its new status as a 'hub' of livestock production gives it an important role in mitigating possible impacts of climate variability on animal health. Animal health may be affected by climate change in four ways: heat-related diseases and stress, extreme weather events, adaptation of animal production systems to new environments, and emergence or re-emergence of infectious diseases, especially vector-borne diseases critically dependent on environmental and climatic conditions. To face these new menaces, the need for strong and efficient Veterinary Services is irrefutable, combined with good coordination of public health services, as many emerging human diseases are zoonoses. Asian developing countries have acute weaknesses in their Veterinary Services, which jeopardises the global surveillance network essential for early detection of hazards. Indeed, international cooperation within and outside Asia is vital to mitigating the risks of climate change to animal health in Asia.

Pinch-induced behavioral inhibition ('clipnosis') in domestic cats.

Research has documented immobilization of rodents, rabbits, guinea pigs and dogs by mechanical means, typically using neck clips or inversion ('animal hypnosis'). In contrast, only a few studies of mechanical immobilization of cats are available, although some success has been reported in the literature. Domestic cats may be effectively immobilized by clips placed along the animal's dorsum. We use the term 'pinch-induced behavioral inhibition' (PIBI) for this behavior because it describes both the method and the response, while avoiding the more anthropomorphic term 'hypnosis'. We investigated the effectiveness of PIBI and its neurological and habituation effects in healthy cats and cats with idiopathic cystitis (IC). Although not all cats were susceptible to PIBI and effectiveness varied among individuals, PIBI was useful for gentle restraint in most cats.

Intracerebroventricularly administered oxytocin attenuated cortisol secretion, but not behavioral responses, during isolation in Holstein steers.

In rodents, intracerebroventricular oxytocin administration attenuated hypothalamo-pituitary-adrenal (HPA) responses and anxiety behavior during stress. We examined the effects of intracerebroventricular injection of oxytocin on isolation-induced stress responses in cattle. In a methodological test, we determined the dosage of oxytocin applied in a main test which did not induce an increase in plasma cortisol concentration or stereotyped behaviors. In a main test, 5 steers aged from 199 to 250 days were assigned to the following three treatments randomly: T1, no isolation after injection of 200 microl of artificial cerebrospinal fluid (aCSF); T2, isolation after aCSF injection; and T3, isolation after 0.5 microg of oxytocin in 200 microl aCSF injection. The isolation was conducted by leaving the experimental steer alone in its stall for one hour while its peers were taken outside. In T2, the isolation induced a rapid increase in plasma cortisol concentration. The maximum %-changes from the pre-isolation value were significantly attenuated by oxytocin injection (T2 vs. T3, p<0.05). The isolation also induced an increase in the frequency (number of occurrences/1 hr isolation) of vocalizations and body orientation changes, and a decrease in the percentage of time spent lying and ruminating. The effect of oxytocin on these behavioral responses to isolation was not apparent. These results indicate that intracerebroventricularly injected oxytocin at low dose attenuated the cortisol response to isolation in steers while the effect on behavior was very small in this experimental condition.

Plasma concentrations of substance P and cortisol in beef calves after castration or simulated castration.

OBJECTIVE: To evaluate plasma concentrations of substance P (SP) and cortisol in calves after castration or simulated castration. ANIMALS: 10 Angus-crossbred calves. PROCEDURES: Calves were acclimated for 5 days, assigned to a block on the basis of scrotal circumference, and randomly assigned to a castrated or simulated-castrated (control) group. Blood samples were collected twice before, at the time of (0 hours), and at several times points after castration or simulated castration. Vocalization and attitude scores were determined at time of castration or simulated castration. Plasma concentrations of SP and cortisol were determined by use of competitive and chemiluminescent enzyme immunoassays, respectively. Data were analyzed by use of repeated-measures analysis with a mixed model. RESULTS: Mean +/- SEM cortisol concentration in castrated calves (78.88+/-10.07 nmol/L) was similar to that in uncastrated control calves (73.01+/-10.07 nmol/L). However, mean SP concentration in castrated calves (506.43+/-38.11 pg/mL) was significantly higher than the concentration in control calves (386.42+/-40.09 pg/mL). Mean cortisol concentration in calves with vocalization scores of 0 was not significantly different from the concentration in calves with vocalization scores of 3. However, calves with vocalization scores of 3 had significantly higher SP concentrations, compared with SP concentrations for calves with vocalization scores of 0. CONCLUSIONS AND CLINICAL RELEVANCE: Similar cortisol concentrations were measured in castrated and control calves. A significant increase in plasma concentrations of SP after castration suggested a likely association with nociception. These results may affect assessment of animal well-being in livestock production systems.