Amphibians, like other vertebrates, respond to acute stressors by releasing glucocorticoid steroid hormones that mediate physiological and behavioural responses to stress. Measurement of stress hormones provides a potential means to improve the welfare of laboratory animals. For example, manipulations of laboratory housing and procedures combined with measurement of glucocorticoids may identify which conditions are more stressful to animals. This is important because there is very little experimental evidence to guide best practice for welfare in amphibians and other lower vertebrates. We investigated the effect of different methods of euthanasia on the circulating plasma corticosterone levels in the African clawed frog (Xenopus laevis), a model amphibian organism that is frequently used in laboratories. In particular, we investigated the effect of different concentrations and pH of the anaesthetic tricaine methanesulphonate (MS-222). Low concentration and unbuffered (low pH) solutions of MS-222 caused elevated corticosterone levels, but only after the effect of MS-222 treatment on blood fluid volume had been taken into account. The level of disturbance that animals experienced also affected corticosterone levels. Thus, our data suggest that to minimise stress to X. laevis, animals should be euthanised after minimal disturbance and in a 3 g L−1 MS-222 solution, buffered to pH 7. The potential for the improvement of amphibian welfare using corticosterone measures as a tool is discussed.

There is no dietary strategy that has yet been specifically advocated for haemophilia. Therefore, we sought to assess the effect of the Dietary Approaches to Stop Hypertension (DASH) diet in adolescents with haemophilia. In this parallel trial, forty male adolescents with haemophilia were dichotomised into the DASH group or control group for 10 weeks. The serum high sensitivity C-reactive protein, IL-6, complete blood count (CBC), serum glutamic oxaloacetic transaminase, serum glutamic pyruvic transaminase, partial thromboplastin time (PTT), waist circumference (WC), percentage of body fat, fat-free mass and liver steatosis were measured at the beginning and end of the study. Serum vitamin C was measured as a biomarker of compliance with the DASH diet. The DASH diet was designed to include high amounts of whole grains, fruits, vegetables and low-fat dairy products, as well as low amounts of saturated fats, cholesterol, refined grains, sweets and red meat. Serum vitamin C in the DASH group was significantly increased compared with the control (P = 0·001). There was a significant reduction in WC (P = 0·005), fat mass (P = 0·006), hepatic fibrosis (P = 0·02) and PTT (P = 0·008) in the DASH group, compared with the control. However, there were no significant differences regarding other selected outcomes between groups. Patients in the DASH group had significantly greater increase in the levels of erythrocyte, Hb and haematocrit, as compared with the control. Adherence to the DASH diet in children with haemophilia yielded significant beneficial effects on body composition, CBC, inflammation and liver function.

Studies have shown the positive effects of prebiotics on the intestinal absorption of Ca and Fe. The present study evaluated the effect of fructo-oligosaccharide (FOS) supplementation in soya beverage (SB) on absorption mechanisms of Ca and Fe in recently weaned rats. Male Wistar rats were divided into four groups: lactose-free cows’ milk (CM), lactose-free CM with FOS (0·8 g/100 ml) (CMF), SB and soya beverage with FOS (0·8 g/100 ml) (SBF). These rats were euthanised after 1 week of treatment. Organ weight, pH of the caecal content and absorption mechanisms of Ca and Fe were evaluated. The results showed that the weight of the caecal contents increased in the CMF and SBF groups, and the pH of the caecal contents was lower in these groups. The Hb levels of the CMF and SB groups were higher when compared with that of the CM group and lower in relation to the SBF group. The apparent Ca and Fe absorption and apparent Ca retention in the CM group were higher when compared with the SB group, whereas in the CMF group, they were higher in relation to the SBF group. Divalent metal transporter 1 (DMT1) protein expression in the duodenum was higher in the SBF group than in the SB and CMF groups. SB resulted in lower intestinal Ca absorption and higher Hb concentration, despite the lower apparent Fe absorption in relation to CM. Supplementation with FOS provided beneficial effects on Hb and DMT1 protein expression in the duodenum, in addition to improving the absorption process.

Blood parasites such as haemogregarines and haemosporidians have been identified in almost all groups of vertebrates and may cause serious damages to their hosts. However, very little is known about biodiversity of these parasites and their effects on some groups of reptiles such as terrapins. Moreover, the information on virulence from blood parasites mixed infection is largely unknown in reptiles. With this aim, we investigated for the first time the prevalence and genetic diversity of blood parasites from one genus of haemoparasitic aplicomplexan (Hepatozoon) in two populations of Spanish terrapins (Mauremys leprosa), a semi-aquatic turtle from southwestern Europe with a vulnerable conservation status. We also examined the association between mixed blood parasite infection and indicators of health of terrapins (body condition, haematocrit values and immune response). Blood parasite infection with Hepatozoon spp was detected in 46·4% of 140 examined terrapins. The prevalence of blood parasites infection differed between populations. We found two different lineages of blood parasite, which have not been found in previous studies. Of the turtles with infection, 5·7% harboured mixed infection by the two lineages. There was no difference in body condition between uninfected, single-infected and mixed-infected turtles, but mixed-infected individuals had the lowest values of haematocrit, thus revealing the negative effects of blood parasite mixed infections. Immune response varied among terrapins with different infection status, where mixed infected individuals had higher immune response than uninfected or single-infected terrapins.

We investigated the physiology of two closely related albatross species relative to their breeding strategy: black-browed albatrosses (Thalassarche melanophris) breed annually, while grey-headed albatrosses (T. chrysostoma) breed biennially. From observations of breeding fate and blood samples collected at the end of breeding in one season and feather corticosterone levels (fCort) sampled at the beginning of the next breeding season, we found that in both species some post-breeding physiological parameters differed according to breeding outcome (successful, failed, deferred). Correlations between post-breeding physiology and fCort, and links to future breeding decisions, were examined. In black-browed albatrosses, post-breeding physiology and fCort were not significantly correlated, but fCort independently predicted breeding decision the next year, which we interpret as a possible migratory carry-over effect. In grey-headed albatrosses, post-breeding triglyceride levels were negatively correlated with fCort, but only in females, which we interpret as a potential cost of reproduction. However, this potential cost did not carry-over to future breeding in the grey-headed albatrosses. None of the variables predicted future breeding decisions. We suggest that biennial breeding in the grey-headed albatrosses may have evolved as a strategy to buffer against the apparent susceptibility of females to negative physiological costs of reproduction. Future studies are needed to confirm this.

Animals frequently host organisms on their surface which can be beneficial, have no effect or a negative effect on their host. Ectoparasites, by definition, are those which incur costs to their host, but these costs may vary. Examples of avian ectoparasites are chewing lice which feed exclusively on dead feather or skin material; therefore, costs to their bird hosts are generally considered small. Theoretically, many possible proximate effects exist, like loss of tissue or food, infected bites, transmission of microparasitic diseases or reduced body insulation due to loss of feathers, which may ultimately also have fitness consequences. Here, we experimentally examined a possible negative impact of 2 feather-eating louse species (Meropoecus meropis and Brueelia apiastri) on male and female European bee-eaters (Merops apiaster) by removing or increasing louse loads and comparing their impact to a control group (lice removed and immediately returned) after 1 month. A negative effect of chewing lice was found on body mass and sedimentation rate and to a lesser extent on haematocrit levels. Males and females lost more weight when bearing heavy louse loads, and were more susceptible to infestations as indicated by the higher sedimentation rate. Our results further suggest differences in sex-specific susceptibility.

l-Carnitine supplementation can stimulate erythropoiesis, reduce exercise-induced plasma lactate concentrations and decrease post-exercise muscle damage. Next to horses, Greyhounds represent the premier animal racing species and perform short-duration, very high-intensity exercise that has the potential to incur substantial muscle damage. Under resting and standard racing conditions (5/16 mile), we tested the novel hypotheses that l-carnitine supplementation in Greyhounds would: (1) elevate haematocrit at rest and immediately post-exercise; (2) reduce peak post-exercise plasma lactate; and (3) reduce indices of muscle damage (plasma creatine phosphokinase, CPK and aspartate aminotransferase, AST). Six conditioned Greyhounds (30.1 ± 1.6 kg) underwent a randomized placebo-controlled crossover study to determine the effects of 6 weeks of l-carnitine supplementation (100 mg kg− 1 of body weight/day) at rest and following a maximal speed 5/16 mile race. In accordance with our hypotheses, l-carnitine elevated resting and immediately post-race haematocrit (control, 60.1 ± 1.7, l-carnitine, 63.6 ± 1.7; P < 0.05) and reduced peak post-race plasma CPK and AST concentrations (both P < 0.05). Those dogs with the highest peak post-exercise plasma CPK concentrations under placebo conditions evidenced the greatest reduction with l-carnitine supplementation (r = 0.99, P < 0.01). However, contrary to our hypotheses, l-carnitine did not change peak post-exercise plasma lactate concentrations (control, 27.0 ± 2.1, l-carnitine, 27.7 ± 1.3; P>0.05). We conclude that l-carnitine supplementation increases the potential for oxygen transport and reduces plasma indicators of muscle damage, CPK and AST in racing Greyhounds.

Background and objective: The study investigated the effect of acute normovolaemic haemodilution on haemodynamics, blood flow and oxygen transport variables with regard to skeletal muscle tissue oxygenation in a canine model.

Methods: Twenty foxhounds were anaesthetized, mechanically ventilated with 30% oxygen in air and underwent first-step normovolaemic haemodilution with Ringer's lactate solution to haematocrit (Hct) 30 and 25% and second-step acute normovolaemic haemodilution with 6% Hetastarch 70 000/0.5 to Hcts of 20, 15 and 10%. Catheters were inserted into femoral arteries and veins and into the pulmonary artery for measurements of haemodynamics, temperature, and sampling of arterial and mixed-venous blood. A flow probe was placed around the left femoral artery. Skeletal muscle tissue oxygen tension (tPO2) was measured in the gastrocnemius muscle using a stepwise driven polarographic needle probe creating histograms from 200 single tPO2 measurements.

Results: Until a Hct of 25% was reached, the heart rate, mean arterial pressure, global and muscular oxygen delivery and consumption remained constant, while the cardiac index and oxygen extraction ratio were significantly increased when compared with baseline. The median tPO2 was significantly decreased at Hcts 15 and 10%, despite increased cardiac index and regional blood flow. The ratio of tPO2 and cardiac index as a marker for efficiency of acute normovolaemic haemodilution started to decline beyond Hcts of 25% (change of slope).

Conclusions: In acute normovolaemic haemodilution to the level of Hct of 25%, the ratio between tPO2 and cardiac index decreases in the healthy dog, indicating an uneconomic relation at the threshold of Hct of 25%.