Neonatal Calf Diarrhea and Gastrointestinal Microbiota: Etiologic Agents and Microbiota Manipulation for Treatment and Prevention of Diarrhea.

Neonatal calf diarrhea is the leading cause of neonatal morbidity and mortality globally. The changes associated with the gastrointestinal microbiota in neonatal calves experiencing diarrhea and its etiology are not fully understood or completely defined in the literature. Several studies have demonstrated that the fecal microbiota of calves that experience diarrhea substantially deviates from that of healthy age-matched calves. However, one key question remains: whether the changes observed in the bacterial communities (also known as dysbiosis) are a predisposing factor for, or the consequence of, gastrointestinal inflammation caused by the pathogens associated with calf diarrhea. The first objective of this literature review is to present the current information regarding the changes in the fecal microbiota of diarrheic calves and the impact of the pathogens associated with diarrhea on fecal microbiota. Modulation of the gastrointestinal microbiota using pre- and probiotics, colostrum feeding, and fecal microbiota transplantation (FMT) has been used to treat and prevent gastrointestinal diseases in humans and dogs. Although information regarding the use of probiotics for the prevention of diarrhea is available in cattle, little information is available regarding the use of these strategies for treating calf diarrhea and the use of prebiotics or FMT to prevent diarrhea. The second objective of this literature review is to summarize the current knowledge regarding the impact of prebiotics, probiotics, synbiotics, colostrum feeding, and FMT for the treatment and prevention of calf diarrhea.

Clinical presentation, medical management, and outcomes in 35 hospitalized sheep diagnosed with bluetongue virus disease.

BACKGROUND: There is only limited information on the clinical presentation, medical management, and outcomes of hospitalized sheep diagnosed with bluetongue virus (BTV) disease. OBJECTIVES: To describe the signalment, history, clinical signs, clinicopathological findings, medical management, and clinical outcomes of sheep diagnosed with BTV disease. ANIMALS: Thirty-five hospitalized sheep with BTV disease. METHODS: Retrospective case series. Medical records from 1989 to 2021 were evaluated. History, signalment, clinical signs, laboratory test results, treatments, and outcomes were recorded. RESULTS: BTV disease was diagnosed from July to December, with a peak proportion (43%; 15/35) of diagnoses recorded in October. Pyrexia and anorexia, respiratory disease, vasculitis, coronitis and lameness, and ulcerative mucosal lesions were present in 71%, 71%, 66%, 49%, and 22% of sheep, respectively. BTV serotypes 10, 11, 13, and 17 were identified, with serotype 17 (75%) being the most frequent. Management of cases included administration of antimicrobials (89%), anti-inflammatories (77%), IV fluids (60%), vitamins (20%), proton-pump inhibitors (14%), diuretics (9%), and antioxidants (9%). Six ewes were pregnant on presentation, but none aborted. Six (17%) sheep died or were euthanized because of clinical deterioration, whereas 83% were discharged. CONCLUSIONS AND CLINICAL IMPORTANCE: The proportion of sheep that survived BTV disease after treatment was relatively high. Serotyping of BTV is recommended because of the mismatch between frequently identified serotypes and the serotype present in the vaccine.

Total and pathogen-specific serum Immunoglobulin G concentrations in neonatal beef calves, Part 1: Risk factors.

Maternal antibodies, delivered to the calf via colostrum, are crucial to prevent calfhood diseases and death. However, knowledge regarding the factors influencing this transfer of total and specific Immunoglobulin G (IgG) against common enteric and respiratory disease pathogens under current production conditions is sparse. The objectives of this study were to determine risk factors influencing total and pathogen-specific immunoglobulin G (IgG) concentrations against Escherichia coli (E. coli), bovine Rotavirus (BRoV), Cryptosporidium parvum (C. parvum), Bovine Viral Diarrhea Virus type 1 and 2 (BVDV), Parainfluenza Virus Type 3 (PI-3), Bovine Respiratory Syncytial Virus (BRSV), and Bovine Herpesvirus type 1 (BHV-1) in the serum of newborn beef calves. A total of 420 serum samples were collected from 1- to 7-day-old beef calves born on 6 farms in Alberta, Canada. Samples were analyzed by radial immunodiffusion for total IgG concentration and by enzyme-linked immunosorbent assays for pathogen-specific IgG concentrations against E. coli, BRoV, C. parvum, BVDV, PI-3, BRSV, and BHV-1. Multivariable multilevel linear and logistic regression models were built to evaluate dam- and calf-level risk factors associated with total and pathogen-specific IgG concentrations, failed transfer of passive immunity (FTPI; serum IgG < 10 g/L), and inadequate transfer of passive immunity (ITPI; serum IgG < 24 g/L). Farm was included as a random effect in all models to account for clustering at the herd level. Of the 420 calves included in this study, 5% (n = 20) and 18% (n = 75) of calves had FTPI and ITPI, respectively. Receiving colostrum intervention (i.e., being fed colostrum or colostrum product by either bottle or tube) was the most consistent risk factor for low total IgG concentration and significantly increased the odds of FTPI (Odds ratio (OR): 6.1, 95% CI: 2.0-18.9) and ITPI (OR: 4.8, 95% CI: 2.1-10.8). Calves born to cows consistently had higher pathogen-specific IgG concentrations (P < 0.0001), compared to calves born from heifers, and calves born to vaccinated dams had significantly higher BRoV, BVDV, and BHV-1-specific IgG concentrations. Interestingly, E.coli-specific IgG concentrations were associated with dam vaccination only in cows but not in heifers, which was likely due to differing vaccination strategies used. This study highlights the need to review and refine protocols with respect to dam vaccination and colostrum intervention on cow-calf operations.

Total and pathogen-specific serum Immunoglobulin G concentrations in neonatal beef calves, Part 2: Associations with health and growth.

The association of poor transfer of passive immunity (TPI) with negative health outcomes is extensively researched in dairy calves. However, few field studies have examined the effect of total and particularly pathogen-specific Immunoglobulin G (IgG) concentrations on pre-weaning health and growth of beef calves. Hence, the objective of this study was to determine the association of total and pathogen-specific IgG concentrations against selected pathogens associated with neonatal calf diarrhea (NCD) and bovine respiratory disease (BRD) and the odds of pre-weaning treatments, mortality, and the growth of newborn beef calves. A total of 420 serum samples from 1- to 7-day old beef calves born on 6 farms in Alberta, Canada, were available for this observational study. Serum samples were analyzed by radial immunodiffusion for total IgG concentration and by enzyme-linked immunosorbent assays for pathogen-specific IgG concentrations against Escherichia coli (E. coli), bovine Rotavirus (BRoV), Cryptosporidium parvum (C. parvum), Bovine Viral Diarrhea Virus (BVDV), Parainfluenza Virus Type 3 (PI-3), Bovine Respiratory Syncytial Virus (BRSV), and Bovine Herpesvirus Type 1 (BHV-1). Data about the individual dam- and calf-level risk factors, calf treatments, mortality, and birth and weaning weights were collected. Multivariable multilevel logistic and linear regression models were built to evaluate the association of total and pathogen-specific IgG concentrations with the odds of mortality and average daily gain (ADG), respectively, while their association with the odds of pre-weaning treatment was established by univariable logistic regression analysis. The odds of calves with IgG concentrations < 10 g/L of getting treated (OR 7.9, 95 % CI 2.7-23.7) and dying (OR: 18.5, 95 % CI: 3.7-93.4) were higher than for their counterparts (P < 0.0001). Calves with IgG concentrations < 24 g/L also had higher odds of dying (OR: 10.1, 95 % CI: 2.6-40.2) and had lower ADG (-0.09 kg, SE: 0.03, P < 0.002) than calves with IgG concentrations ≥ 24 g/L. Higher BVDV-specific IgG concentration was protective against mortality (OR: 0.97, 95 % CI: 0.96-0.99, P < 0.001). This study highlights the negative association of inadequate TPI with health and growth of beef calves and thus, the need to refine protocols for dam vaccination and colostrum intervention in cow-calf operations to meet these higher IgG targets.

Effect of time of sample collection after onset of diarrhea on fecal microbiota composition of calves.

BACKGROUND: The effect of time of sample collection after onset of diarrhea on the fecal microbiota composition of calves is unknown. OBJECTIVE: Compare the fecal microbiota of calves with diarrhea onset on the day of sampling (D <24h), and calves having had diarrhea for >24 to 48 hours (D 24-48h). ANIMALS: Thirty-one diarrheic calves (20 D <24h and 11 D 24-48h), 3 to 7 days of age. METHODS: Cross-sectional study. Diarrhea was defined as a calf with loose feces or watery feces. Assessment of the fecal microbiota was performed by sequencing of 16S ribosomal RNA gene amplicons. RESULTS: Richness and diversity were not statistically different between D <24h and D 24-48h (P > .05), but bacterial membership and structure differed significantly (AMOVA, P < .001 for both comparisons). Linear discriminant analysis effect size (LefSe) showed an enrichment of Faecalibacterium, Phocaeicola, Lachnospiracea, and Lactobacillus in the feces of D <24h calves, whereas Escherichia/Shigella, Ligilactobacillus, Clostridium_Sensu_Stricto, Clostridium_Incerta_Sedis, and Enterococcus were enriched in the D 24-48h calves. CONCLUSION AND CLINICAL IMPORTANCE: Rapid changes in fecal microbiota occur during the first 48 hours of diarrhea with an enrichment of lactic acid-producing bacteria in D <24h followed by an enrichment in Escherichia/Shigella and Clostridium spp. in D 24-48h. The time from diarrhea onset to sampling appears to affect the bacterial composition. Researchers should standardize times for fecal collection based on the time of diarrhea.

Host innate immune responses and microbiome profile of neonatal calves challenged with Cryptosporidium parvum and the effect of bovine colostrum supplementation.

Introduction: Calves are highly susceptible to gastrointestinal infection with Cryptosporidium parvum (C. parvum), which can result in watery diarrhea and eventually death or impaired development. With little to no effective therapeutics, understanding the host's microbiota and pathogen interaction at the mucosal immune system has been critical to identify and test novel control strategies. Methods: Herein, we used an experimental model of C. parvum challenge in neonatal calves to describe the clinical signs and histological and proteomic profiling of the mucosal innate immunity and microbiota shifts by metagenomics in the ileum and colon during cryptosporidiosis. Also, we investigated the impact of supplemental colostrum feeding on C. parvum infection. Results: We showed that C. parvum challenged calves experienced clinical signs including pyrexia and diarrhea 5 days post challenge. These calves showed ulcerative neutrophil ileitis with a proteomic signature driven by inflammatory effectors, including reactive oxygen species and myeloperoxidases. Colitis was also noticed with an aggravated mucin barrier depletion and incompletely filled goblet cells. The C. parvum challenged calves also displayed a pronounced dysbiosis with a high prevalence of Clostridium species (spp.) and number of exotoxins, adherence factors, and secretion systems related to Clostridium spp. and other enteropathogens, including Campylobacter spp., Escherichia sp., Shigella spp., and Listeria spp. Daily supplementation with a high-quality bovine colostrum product mitigated some of the clinical signs and modulated the gut immune response and concomitant microbiota to a pattern more similar to that of healthy unchallenged calves. Discussion: C. parvum infection in neonatal calves provoked severe diarrheic neutrophilic enterocolitis, perhaps augmented due to the lack of fully developed innate gut defenses. Colostrum supplementation showed limited effect mitigating diarrhea but demonstrated some clinical alleviation and specific modulatory influence on host gut immune responses and concomitant microbiota.

The impact of freezing and multiple freeze-thaw cycles on Brix refractometry estimates of immunoglobulin concentration in beef cattle colostrum.

Evaluation of immunoglobulin G (IgG) concentration in colostrum is important to guide on-farm management. Studies have shown that digital Brix refractometry accurately estimates colostrum IgG concentration in both dairy and beef cattle colostrum. Colostrum is often frozen in both clinical and research settings. The implications of this freezing on the accuracy of Brix refractometry measurements are largely unknown. The first objective of this study was to evaluate the agreement between digital Brix percentage measurements of IgG in beef cattle colostrum taken before and after different durations of freezing. The second objective was to evaluate the effects of multiple freeze-thaw (FT) cycles on Brix percentage measurements of IgG in beef cattle colostrum. There was good agreement between Brix percentages in fresh colostrum and after short (2 to 8 d), medium (4 to 7 mo), and long (3 y) periods of freezing (concordance correlation coefficient: 0.95, 0.96, and 0.96, respectively). Although there was no significant change in mean Brix percentages over 2 FT cycles (P > 0.05), mean Brix percentages decreased with 3 FT cycles (P = 0.017). Samples from the fourth and fifth FT cycles were observably coagulated, and these measurements were therefore deemed inaccurate. Data from this study indicate that freezing had minimal impact on digital Brix refractometer estimates of IgG concentration in beef cattle colostrum, but that samples stored for future testing should not undergo more than 2 FT cycles.

L'évaluation de la concentration d'immunoglobuline G (IgG) dans le colostrum est importante pour guider la gestion à la ferme. Des études ont montré que la réfractométrie Brix numérique estime avec précision la concentration d'IgG du colostrum dans le colostrum des bovins laitiers et de boucherie. Le colostrum est souvent congelé dans les milieux cliniques et de recherche. Les implications de cette congélation sur la précision des mesures de réfractométrie Brix sont largement inconnues. Le premier objectif de cette étude était d'évaluer la concordance entre les mesures numériques du pourcentage de Brix d'IgG dans le colostrum de bovins de boucherie prises avant et après différentes durées de congélation. Le deuxième objectif était d'évaluer les effets de plusieurs cycles de congélation-décongélation (FT) sur les mesures du pourcentage Brix d'IgG dans le colostrum de bovins de boucherie. Il y avait un bon accord entre les pourcentages de Brix dans le colostrum frais et après des périodes de congélation courtes (2 à 8 jours), moyennes (4 à 7 mois) et longues (3 ans) (coefficient de corrélation de concordance : 0,95, 0,96 et 0,96, respectivement). Bien qu'il n'y ait pas eu de changement significatif dans les pourcentages moyens de Brix sur deux cycles FT (P > 0,05), les pourcentages moyens de Brix ont diminué avec trois cycles FT (P = 0,017). Les échantillons des quatrième et cinquième cycles FT étaient coagulés de manière observable, et ces mesures ont donc été jugées inexactes. Les données de cette étude indiquent que la congélation a eu un impact minimal sur les estimations du réfractomètre numérique Brix de la concentration d'IgG dans le colostrum de les bovins de boucherie, mais que les échantillons stockés pour les tests futurs ne doivent pas subir plus de deux cycles FT.(Traduit par Docteur Serge Messier).

Effect of plasma transfusion on serum amyloid A concentration in healthy neonatal foals and foals with failure of transfer of passive immunity.

BACKGROUND: Anecdotal evidence suggests plasma transfusions increase serum amyloid A (SAA) concentrations in healthy neonatal foals making this marker of inflammation inappropriate for therapeutic decision making in such animals. HYPOTHESIS/OBJECTIVES: Administration of hyperimmune fresh frozen plasma (FFP) increases SAA concentration in healthy foals and in foals with failure of transfer of passive immunity (FTPI). ANIMALS: Eighty-six healthy foals. METHODS: Prospective cohort study. Foals <24 hours of age receiving plasma transfusion for treatment of FTPI (serum immunoglobulin G [IgG] concentrations <8 g/L; n = 17) or as a preventative measure for Rhodococcus equi infection (IgG >8 g/L; n = 33) were enrolled. A healthy nontransfused group of foals (IgG >8 g/L; n = 21) also was included. Serum amyloid A concentration was determined before (t0h) and after (t24h) administration of FFP. Changes in blood SAA concentration were assessed using linear regression models. RESULTS: No statistical differences were found in SAA concentration at t0h or t24h among the 3 groups (P > .05, for all comparisons). The variation in SAA concentration before (t0h) and after (t24h) plasma transfusion showed that administration of FFP was not associated with the changes in SAA concentration (P > .05). An association between SAA concentration at t0h and at 24 hours (P < .05) was identified, where foals with higher SAA concentration at t0h also had higher SAA concentration at t24h. CONCLUSIONS AND CLINICAL IMPORTANCE: Administration of FFP to newborn foals was not associated with changes in SAA concentration.

Risk factors for, frequency, and type of complications after temporary tube cystostomy in goats, sheep, and pigs.

OBJECTIVE: To describe risk factors for as well as type and frequency of complications, time to occurrence of complications, and mortality after temporary tube cystostomy alone or in combination with other surgical procedures, in goats, sheep, and pigs. STUDY DESIGN: Retrospective case series. ANIMALS: One hundred thirteen goats, eight sheep, and six pigs. METHODS: Medical records of goats, sheep, and pigs that underwent temporary tube cystostomy alone or in combination with urethrotomy at the Veterinary Medicine Teaching Hospital, University of California-Davis from January 1, 2007 to December 31, 2018 were reviewed. Signalment, presurgical and surgical procedures, urolith type and location, and postoperative complications and mortality were analyzed. In goats, logistic regression analysis was performed to determine association between the presence of complications or mortality as well as candidate predictor variables. RESULTS: Postoperative complications were reported in 54.9% (62/113) of goats, 37.5% (3/8) of sheep, and 50% (3/6) of pigs. The most frequent type of complication was reobstruction. Median (range) time to first complication was 171 days (1-2247) for goats. In sheep and pigs, the range for time to re-presentation for complications was 11 to 37 and 17 to 1182 days, respectively. Proportions of nonsurvivors after tube cystostomy were 48.4% (30/62) goats, 1/8 sheep and 2/6 pigs. No association between examined predictor variables and the occurrence of complications or mortality was detected. CONCLUSION: The risk of complications and mortality after temporary tube cystostomy was high, especially in goats. CLINICAL SIGNIFICANCE: Owners should be made aware of the high complication rate after temporary tube cystostomy alone or in combination with other procedures. Investigation of alternative approaches to manage obstructive urolithiasis in goats, sheep, and pigs should be considered.

Evaluation of 3 refractometers to determine transfer of passive immunity in neonatal beef calves.

BACKGROUND: Refractometry is used to assess transfer of passive immunity (TPI), but studies evaluating different refractometers and appropriate thresholds for recommended target immunoglobulin G (IgG) concentrations for beef calves are limited. OBJECTIVES: To evaluate test performance of digital (DSTP) and optical (OSTP) serum total protein (STP) refractometers and a digital Brix (DBRIX) refractometer for assessment of passive immunity in beef calves. ANIMALS: A total of 398 beef calves from 6 herds, 1 to 7 days of age. METHODS: Serum IgG concentration was estimated by DSTP, OSTP, and DBRIX, and measured by radial immunodiffusion (RID). Correlation coefficients (r) among results were calculated. Optimal STP and Brix thresholds for identification of IgG <10, <16, and <24 g/L were determined using interval likelihood ratios. Refractometer performance and agreement were assessed using areas under the curve (AUC), diagnostic test characteristics, Cohen's kappa (κ), and Bland-Altman analysis. RESULTS: Refractometer results were highly correlated with RID (r = 0.82-0.91) and with each other (r = 0.91-0.95), and overall test performance was excellent (AUC = 0.93-0.99). The STP concentrations of ≤5.1, ≤5.1, and ≤5.7 g/dL and Brix percentages of ≤7.9%, ≤8.3%, and ≤8.7% indicated IgG concentrations <10, <16, and <24 g/L, respectively. Agreement of refractometers with RID was variable (κ = 0.46-0.80) and among refractometers was substantial (κ = 0.62-0.89). CONCLUSIONS AND CLINICAL IMPORTANCE: All refractometers showed good utility as monitoring tools for assessment of TPI in beef calves.

Efficacy of sodium iodide for prevention of respiratory disease in preweaned dairy calves.

OBJECTIVE: To determine the pharmacokinetics of sodium iodide (NaI) following oral administration to preweaned dairy calves, and to assess the efficacy of NaI for prevention of bovine respiratory disease (BRD) in preweaned calves at a commercial calf-raising facility. ANIMALS: 434 healthy preweaned dairy calves. PROCEDURES: In the first of 2 experimental trials, each of 7 calves received NaI (20 mg/kg, PO) once. Blood and nasal fluid samples were collected at predetermined times before (baseline) and for 72 hours after NaI administration for determination of iodine concentrations. Pharmacokinetic parameters were determined by noncompartmental analysis. In the second trial, 427 calves at a calf-raising facility were randomly assigned to receive NaI (20 mg/kg, PO, 2 doses 72 hours apart; n = 211) or serve as untreated controls (216). Health outcomes were compared between the 2 groups. RESULTS: For all 7 calves in the pharmacokinetic trial, the iodine concentration in both serum and nasal fluid samples was significantly increased from the baseline concentration and exceeded the presumed therapeutic iodine concentration (6.35 µg/mL) throughout the sampling period. In the on-farm trial, the odds of being treated for BRD before weaning for NaI-treated calves were twice those for control calves (OR, 2.04; 95% CI, 1.38 to 3.00). CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that, although oral administration of NaI (20 mg/kg) to preweaned dairy calves achieved iodine concentrations presumed to be effective in both serum and nasal fluid, it was not effective for prevention of BRD in preweaned calves at a commercial calf-raising facility.

Assessment of Brix refractometry to estimate immunoglobulin G concentration in beef cow colostrum.

BACKGROUND: Brix refractometry can be used to assess colostral immunoglobulin G (IgG) concentration, but studies identifying Brix percentages to detect high- and low-IgG colostrum are lacking for beef cows and interlaboratory agreement is unknown. OBJECTIVES: Evaluate Brix refractometer performance and interlaboratory agreement for assessing beef cow colostrum IgG concentration, including determination of thresholds to identify colostrum containing IgG concentrations <100 g/L and ≥150 g/L. ANIMALS: Beef cows (n = 416) from 11 cow-calf operations in Alberta, Canada. METHODS: Colostral IgG concentrations were measured using radial immunodiffusion (RID) and estimated by Brix refractometry for this retrospective study. Spearman correlation coefficients were assessed between RID and Brix refractometry. Likelihood ratios and misclassification cost-term analysis were used to determine optimal Brix percentages for detecting colostrum containing IgG concentrations <100 g/L and ≥150 g/L. Concordance correlation coefficient (CCC) and Bland-Altman analyses were performed for Brix percentages obtained at 3 different laboratories. RESULTS: Brix percentages obtained at 3 laboratories were positively correlated with IgG results (r = 0.72, 0.68, and 0.76, respectively). Colostrum Brix percentages of <24% and ≥30% were optimal for indicating IgG concentrations of <100 g/L and ≥150 g/L, respectively. Interlaboratory agreement was substantial, with CCC ranging from 0.89 to 0.96 and Bland-Altman analysis showing small mean differences (-1.2% to 0.09% Brix) and narrow limits of agreements (-4.8% to 2.4% Brix) among laboratories. CONCLUSIONS AND CLINICAL IMPORTANCE: Brix refractometry shows good potential for reliably estimating IgG concentrations in beef cow colostrum across multiple laboratories and can be recommended to aid colostrum management decisions on farms.

Vaccinating Calves in the Face of Maternal Antibodies: Challenges and Opportunities.

A growing body of evidence has shown that calves can mount an immune response when vaccinated in the face of maternal antibodies (IFOMA), albeit inconsistently and often in ways that differ from seronegative calves or older cattle. Several previous reviews have endeavored to explain bovine neonatal immunology and have documented the issue of vaccinating young calves. However, as preweaning vaccination becomes more common in both beef and dairy production systems, so too has research on the impacts of such vaccination programs. This article aims to briefly review the challenges and opportunities for vaccinating calves IFOMA.

Healthy n-6/n-3 fatty acid composition from five European game meat species remains after cooking.

BACKGROUND: Intensive farming of livestock along with recent food scandals and consumer deception have increased awareness about risks for human nutrition. In parallel, the demand for meat obtained under more natural conditions from animals that can freely forage has largely increased. Interestingly, the consumption of game meat has not become more common despite its excellent quality and content of polyunsaturated fatty acids (PUFAs). CONTEXT AND PURPOSE: We addressed the question if game meat fatty acid composition is modified through kitchen preparation. By analysing muscle fatty acid (FA) composition (polar and total lipids) of five European game species in a raw and a processed state, we aimed to quantify the proportion of PUFA that are oxidised and hydrogenated during processing. All game meat species originated from local hunters and free-living individuals. To mimic a realistic situation a professional chef prepared the meat samples with gentle use of heat in a standardised way. RESULTS: Expectedly, the overall content of polyunsaturated fatty acids declined during the cooking process but the decrease size was <5% and the nutritiously most important n-3/n-6 ratio was not affected by processing (F1,54 = 0.46; p = 0.5). Generally, our samples contained species-specific high PUFA and n-3 FA contents but we point out that differentiating between species is necessary. CONCLUSION: Game meat thus provides a healthy meat source, as cooking does not substantially alter its favourable fatty acid composition. Further research is needed to elucidate species-specific differences and the role of habitat quality and locomotion for tissue composition.