Transportation stress causes significant changes in physiological responses in goats; however, studies exploring the transcriptome of stress are very limited. The objective of this study was to determine the differential gene expressions and related pathways in the blood samples using RNA-seq procedure in Spanish goats subjected to different durations of transportation stress. Fifty-four male Spanish goats (8-mo old; BW = 29.7 ± 2.03 kg) were randomly subjected to one of three treatments (TRT; n = 18 goats/treatment): (1) transported for 180 min, (2) transported for 30 min, or (3) held in pens (control). Blood samples were collected before and after treatment for stress hormone, metabolite, and transcriptomic analysis. RNA-seq technology was used to obtain the transcriptome profiles of blood. Analysis of physiological data using SAS showed that plasma cortisol concentrations were higher(P < 0.01) in 180 min and 30 min groups compared to the control group. Enrichment analysis of DEGs related to transportation stress through Gene Ontology and KEGG databases revealed that the differentially expressed genes related to inflammatory pathways, caspases, and apoptosis such asIL1R2, CASP14, CD14, TLR4, andMAPK14were highly enriched in the transported group of goats compared to non-transported goats. Stress in goats leads to a sequence of events at cellular and molecular levels that causes inflammation and apoptosis.

Social isolation can increase distress in goats, particularly when they cannot maintain visual contact with conspecifics. This experiment was conducted to determine the behavioral and physiological responses in goats during isolation with or without visual contact with conspecifics. Male Spanish goats (uncastrated, 8-mo old, average weight 29.4 ± 0.59 kg) were randomly assigned to a control (CO) group with no isolation or to one of four isolation treatment (TRT) pens (1.5 × 1.5 m) with: (1) open grill panels but with no visual contact with conspecifics (IO), (2) covered grill to prevent visual contact (IC), (3) open grill with visual contact (IV), or (3) covered grill with a 30 × 30 cm window to allow visual contact (IW) for 90 min of social isolation (n = 12 goats/TRT). Blood samples were collected at 0, 30, 60, and 90 min (Time) from isolated and control goats. The experiment was repeated one week later using the same animals, with each goat being subjected to the same isolation treatment the second time to study the effect of prior exposure to isolation. Friedman’s Two-Way ANOVA by Ranks Test in SAS showed that the median frequency of vocalization (rank score) in goats was high in IO group, low in IV and IW groups, and intermediate in IC group (P < 0.01). Vocalization rank score was also higher (P < 0.01) during the first 30 min of isolation in goats. Median frequency of visual contact was higher in the IW group than in the IV group (P < 0.01). Frequency of climbing behavior was high in IC and IO groups, low in IV group, and intermediate in IW group (P < 0.01). Repeated Measures Analysis using GLM procedures in SAS revealed that plasma cortisol and glucose concentrations tended (P < 0.1) to be the highest in IO group than in CO, IC, IV, and IW groups. Cortisol levels were also higher (Time; P < 0.05) at 0 and 90 min compared to 30 and 60 min. Norepinephrine concentrations decreased (P < 0.05) with Time, and plasma non-esterified fatty acid (NEFA) levels were affected by TRT × Time interaction (P < 0.01). Overall, epinephrine, norepinephrine, glucose, and NEFA concentrations were lower (P < 0.01) and cortisol concentrations and lymphocyte counts higher (P < 0.01) when goats were exposed to isolation the second time. The results showed that goats with no visual contact with conspecifics during social isolation had greater physiological stress responses and spent more time vocalizing or trying to escape the pen, which may indicate distress.

Feeding condensed tannin (CT)-containing diets such as sericea lespedeza (Lespedeza cuneata) and reducing stress have been reported to improve meat quality and food safety in goats. In a completely randomized design with split-plot, thirty-six uncastrated male Spanish goats were assigned to 3 dietary treatments (n = 12/treatment): ground ‘Serala’ sericea lespedeza hay (SER), bermudagrass (Cynodon dactylon) hay (BG), or bermudagrass hay—dewormed goats (BG-DW; Control) at 75% of intake, with a corn-based supplementation (25%) for 8 weeks. Prior to slaughter, goats were either transported for 90 min to impose stress or held in pens. Basophil counts were lower (p < 0.01) in the SER group compared to BG or BG-DW groups suggesting a better anti-inflammatory capacity due to polyphenols in the SER diet. Compared to BG-DW group, cortisol level was higher (p < 0.05) and norepinephrine was lower (p < 0.05) in the SER group. The SER group had the lowest aerobic plate counts (APC) in both rumen and rectum (p < 0.01). Longissimus dorsi muscle initial pH was not affected by diet or stress. Feeding sericea hay to goats may have beneficial effects, such as enhanced antioxidant and anti-inflammatory properties during stress and reduced gut microbial counts, without changing meat quality characteristics.

The effects of high-condensed tannin (CT) diet combined with preslaughter stress have not been studied at the metabolome level in goats. This study was conducted to determine the effects of feeding sericea lespedeza (SL; Lespedeza cuneata), a high-CT legume, and transportation stress on plasma metabolome in goats. Uncastrated male Spanish goats (age = 8 months; BW = 26.0 ± 0.48 kg) were either fed ground ‘Serala’ SL hay (SER), bermudagrass (Cynodon dactylon) hay (BG), or bermudagrass hay-dewormed goats (BG-DW; Control) at 75% of intake, with a corn-based supplementation (25%) for 8 weeks (n = 12/Diet). At the end of the trial, goats were subjected to one of two stress treatments (ST): transported for 90 min to impose stress (TS) or held in pens (NTS) before slaughtering, in two replicates. Live and carcass weights, and blood samples were collected at 0, 30, 60 and 90 min of transportation or holding time (Time). The data were analyzed using MIXED procedures in SAS and metabolomics data were analyzed using the R software. When measured after ST, SER group had the lowest body weight (P < 0.05) among the three diet groups. Carcass weights were high in the BG-DW, low in SER, and intermediate in BG group. Plasma creatine concentrations decreased over Time (P < 0.01) in the TS goats in all diet groups. Meat crude protein percentages were higher (P < 0.05) in SER (22.5 ± 0.22) and BG-DW (22.3 ± 0.22) groups compared to the BG group (21.6 ± 0.22). At the metabolome level, SER group had the lowest (P < 0.05) glycine, alanine, threonine, taurine, trans-hydroxyproline, methionine, and histidine concentrations and highest (P < 0.01) lysine and citrulline concentrations among the Diet groups. Butyric acid, concentration was higher (P < 0.05) in the SER group compared to BG group. Eight medium- and long-chained acylcarnitines were higher (P < 0.05) in the BG-DW group than SER or BG groups. In general, amino acid levels decreased and acylcarnitine increased with Time (P < 0.05) in all groups. Sericea diet can be beneficial in enhancing stress coping abilities in goats due to elevated butyrate, lysine, and citrulline levels; however, SER resulted in lower energy level in goats compared to BG or BG-DW groups. Fatty acid metabolism is the main energy pathway in all groups during prolonged stress. Inclusion of certain varieties of SL in the diet must be carefully controlled to prevent possible negative effect.

Studies exploring the transcriptome of stress and its effects on meat quality are very limited, particularly in goats. Fifty-four male Spanish goats (8-mo old; BW = 29.7 ± 2.03 kg) were randomly subjected to one of three treatments (TRT; n = 18 goats/treatment): (1) transported for 180 min, (2) transported for 30 min, or (3) held in pens (control) to analyze the transcriptome of stress and meat quality in goats using RNA-seq technology. Blood samples were collected before and after treatment, and meat samples were collected after humane slaughter for stress hormone, meat quality (Longissimus dorsi), and transcriptomic analysis. Plasma epinephrine concentrations were higher (P < 0.01) in 180 min and 30 min groups compared to the control group; however, norepinephrine concentrations were not affected by the treatment. Muscle glycogen concentrations (15 min postmortem) were lower (P < 0.01) in both 30 min and 180 min groups compared to the control group. Calpastatin levels were higher (P < 0.01) in 180 min and 30 min groups than the control group. Warner–Bratzler shear force values of loin chops were the highest in the 180 min group (4 ± 0.15, kg), lowest in the control group (3.51 ± 0.10, kg), and intermediate in the 30 min group (3.78 ± 0.09, kg;P < 0.01) both at day 1 and day 6 aging time. Additionally, desmin levels of day 6 samples were lowest in the control group, highest in 180 min group, and intermediate in 30 min group (P < 0.05). RNA-seq results showed that a total of 10,633 genes were differentially expressed (5194 up regulated; 5439 down regulated) among all comparisons (blood and day 1 and day 6 muscle samples). Among these differentially expressed genes (DEGs), KLF9, AMPK, FOXO3, PTX3, GADD45, PTPN1, CASP7, MAPK4, HSPA12A, and JAK-STAT were probably associated with the effects of stress on skeletal muscle proteins and involved in biological process such as cellular response to corticosteroid stimulus, endoplasmic reticulum stress, insulin resistance, DNA repair, apoptosis, MAPK cascade and regulation of proteolysis. The KEGG analysis revealed that AMPK and JAK-SAT signaling pathways and autophagy were among the top 20 enriched pathways in our treatment comparisons. The results provide an understanding of the genes and pathways involved in stress responses and related changes in postmortem muscle metabolism and meat quality characteristics in goats.

This experiment was conducted to determine the effects of habituation to livestock trailers on stress responses in goats transported for long periods. Intact male Spanish goats (12-month old; BW = 31.6 ± 0.34 kg; N = 168) were separated into two treatment (TRT) groups and maintained on two different paddocks. Concentrate supplement was fed to one group inside two livestock trailers (5.0 × 2.3 m each; habituated group, H), while the other group received the concentrate supplement, but not inside the trailers (non-habituated, NH). After 4 weeks of habituation period, goats were subjected to a 10-h transportation stress in four replicates (n = 21 goats/replicate/TRT). Blood samples were collected by a trained individual by jugular venipuncture into vacutainer tubes before loading (Preload), 20 min after loading (0 h), and at 2-h intervals thereafter (Time) for analysis of stress responses. There was a tendency for a TRT effect (p < 0.1) on tyramine and metanephrine concentrations. Phenylethylamine and 5-methoxytryptamine concentrations were significantly greater (p < 0.05) in the H group compared to the NH group. Both dopamine and 5-methoxytryptamine concentrations decreased (p < 0.05) with transportation time; however, TRT × Time interaction effects were not significant. Habituation to trailers may be beneficial in mood and energy stabilization in goats during long-distance transportation.