The effects of zinc amino acid complex on biomarkers of gut integrity, inflammation, and metabolism in heat-stressed ruminants.

Study objectives were to evaluate the effects of replacing 40 mg/kg of dietary Zn from Zn sulfate (ZS) with Zn amino acid complex (ZA; Zinpro Corporation, Eden Prairie, MN) on inflammation and intestinal integrity in heat-stressed and pair-fed (PF) ruminants. Forty Holstein steers (173.6 ± 4.9 kg) were randomly assigned to 1 of 5 dietary-environmental treatments: (1) thermoneutral (TN) ad libitum with 75 mg/kg of dry matter (DM) ZS (ZSCON); (2) TN pair-fed with 75 mg/kg DM ZS (ZSPF); (3) TN pair-fed with 40 mg/kg DM ZA and 35 mg/kg DM ZS (ZAPF); (4) heat stress (HS) ad libitum with 75 mg/kg DM ZS (ZSHS); and (5) HS ad libitum 40 mg/kg DM ZA and 35 mg/kg DM ZS (ZAHS). Before study initiation, calves were fed their respective diets for 21 d. Following the pre-feeding phase, steers were transferred into environmental chambers and were subjected to 2 successive experimental periods. During period 1 (5 d), all steers were fed their respective diets ad libitum and housed in TN conditions (20.2 ± 1.4°C, 30.4 ± 4.3% relative humidity). During period 2 (6 d), ZSHS and ZAHS steers were exposed to cyclical HS conditions (27.1 ± 1.5°C to 35.0 ± 2.9°C, 19.3 ± 3.5% relative humidity), whereas the ZSCON, ZSPF, and ZAPF steers remained in TN conditions and were fed ad libitum or pair-fed relative to their ZSHS and ZAHS counterparts. Overall, steers exposed to HS had markedly increased rectal temperature (0.83°C), respiration rate (26 breaths per min), and skin temperature (8.00°C) relative to TN treatments. Rectal temperature from ZAHS steers was decreased (0.24°C) on d 4 to 6 of HS relative to ZSHS steers. Regardless of diet, HS decreased DMI (18%) relative to ZSCON steers. Circulating glucose from HS and PF steers decreased (16%) relative to ZSCON steers. Heat stress and nutrient restriction increased circulating nonesterified fatty acids 2- and 3-fold, respectively, compared with ZSCON steers. Serum amyloid A increased ~2-fold in PF relative to ZSCON and HS steers. We detected no treatment effect on blood pH; however, ZAHS steers had increased HCO3 relative to ZSHS. Relative to ZSHS, ZAHS steers had increased jejunum villi height (25%), a tendency for increased ileum villi height (9%), and decreased duodenal villi width (16%). In summary, ZA supplementation has some beneficial effects on thermal indices, intestinal architecture characteristics, and biomarkers of leaky gut in heat-stressed steers, indicative of an ameliorated heat load, and thus may be a nutritional strategy to minimize negative consequences of HS.

Assessment of water source availability and quality for small ruminant consumption in the Northern Badia region of Jordan.

BACKGROUND AND AIM: Water is the most important nutrient for the production of healthy livestock. Water scarcity bottlenecks livestock production in arid and semi-arid regions, particularly during the dry season. This study aimed to assess water availability and quality for small ruminants, and to identify major challenges of meeting their water requirements in two major small ruminant production systems in Jordan. MATERIALS AND METHODS: Transhumant and sedentary production systems in the Northern Badia region of Jordan were the focus of this study. A questionnaire was distributed to 120 sheep and goat farmers (62 transhumant farmers and 58 sedentary farmers) and a water quality assessment was completed. RESULTS: Results showed that the two production systems varied their water source use seasonally. Water provision was perceived as one of the major constraints for Bedouins, particularly during the dry season in transhumant production systems, when longer distances to water sources and the high costs of fetching water daily aggravated the problem. The mean distance and travel times to the boreholes were less in the sedentary system. Watering frequency was significantly lower in the transhumant system compared to the sedentary system (p<0.05). Although the values of water quality parameters complied with guidelines for livestock consumption, low water quality was the main concern expressed by some of the survey households. CONCLUSION: Technical support to properly design, manage, and rehabilitate surface water harvesting systems is required for the sustainable use of water resources in the study region. Moreover, systematic water quality monitoring is necessary to ensure its suitability for livestock use. Further investigations on the microbiological quality of water and its effect on the health and performance of livestock are recommended.

Short communication: Ketosis, feed restriction, and an endotoxin challenge do not affect circulating serotonin in lactating dairy cows.

Circulating serotonin (5-hydroxytryptamine; 5-HT) appears to be associated with various energetic disorders and hypocalcemia during the transition period. The objective of this study was to evaluate the effects of ketosis, feed restriction (FR), and endotoxin challenge (models in which energetic and calcium metabolism are markedly altered) on circulating 5-HT in lactating Holstein cows. Blood samples were obtained from 3 separate experiments; circulating ß-hydroxybutyrate (BHB), nonesterified fatty acids (NEFA), and glucose were measured in all 3 experiments, whereas ionized calcium (iCa2+) was measured only in the endotoxin challenge. In the ketosis study, blood samples from cows clinically diagnosed with ketosis (n = 9) or classified as healthy (n = 9) were obtained from a commercial dairy farm at d -7, 3, and 7 relative to calving. Ketosis was diagnosed using a urine-based test starting at 5 d in milk. There was no effect of health status on circulating 5-HT and no association between 5-HT and BHB, NEFA, or glucose; however, 5-HT concentrations progressively decreased following calving. In the FR experiment, mid-lactation cows were either fed ad libitum (n = 3) or restricted to 20% of their ad libitum intake (n = 5) for 5 d. There were no FR effects on circulating 5-HT, nor was FR correlated with energetic metabolites. In the immune activation model, mid-lactation cows were intravenously challenged with either lipopolysaccharide (LPS; 1.5 µg/kg of BW; n = 6) or sterile saline (control; n = 6). Administering LPS decreased (56%) blood iCa2+ but had no effect on circulating 5-HT, nor was there a correlation between circulating 5-HT and NEFA, BHB, or iCa2+. Circulating 5-HT tended to be positively correlated (r = 0.54) with glucose in Holstein cows administered LPS. In summary, in contrast to expectations, circulating 5-HT was unaffected in models of severely disturbed energetic and Ca2+ homeostasis.

Anticoccidial effect of apple cider vinegar on broiler chicken: an organic treatment to measure anti-oxidant effect.

The objective of this study was to investigate the anticoccidial effect of apple cider vinegar added to drinking water with the anticoccidial effect of amprolium to feed broiler chicken. The study has adopted an observational approach to evaluate the anticoccidial effect of apple cider vinegar on broiler chicken. The antioxidative changes were measured adding natural apple cider vinegar to drinking water. Four hundred and fifty broiler chickens were purchased from the local market and distributed into three groups (T+vc: positive control, T-vc: negative control Tv: apple cider vinegar) with 150 chickens in each group. The three groups were further replicated into 3 blocks each containing 50 chickens. The groups were fed balanced diet, amprolium was added to the feed of positive control group, and apple cider vinegar was added to the water of Tv group. Measurements of the different variables were started from week 3, at the end of each week 3 birds were chosen randomly, blood samples were collected via the wing vein, and fecal oocysts were counted from intestinal contents of each individual bird using the McMaster technique. Broiler in the control groups T+ve and T-ve showed clinical signs of coccidiosis (blood in feces) and the number of coccidial oocytes in feces increased with time. In the vinegar group, no clinical signs of coccidiosis were observed. Concentrations of total antioxidants and catalase enzyme activity significantly increased (p≤0.05); while malondialdehyde concentration significantly decreased (p≤0.05).

Effects of heat stress and insulin sensitizers on pig adipose tissue.

Heat stress (HS) negatively impacts several swine production variables, including carcass fat quality and quantity. Pigs reared in HS have more adipose tissue than energetically predicted, explainable, in part, by HS-induced hyperinsulinemia. Study objectives were to evaluate insulin's role in altering fat characteristics during HS via feeding insulin-sensitizing compounds. Forty crossbred barrows (113 ± 9 kg BW) were randomly assigned to one of five environment by diet treatments: 1) thermoneutral (TN) fed ad libitum (TNAL), 2) TN and pair-fed (TNPF), 3) HS fed ad libitum (HSAL), 4) HS fed ad libitum with sterculic oil (SO) supplementation (HSSO; 13 g/d), and 5) HS fed ad libitum with dietary chromium (Cr) supplementation (HSCr; 0.5 mg/d; Kemin Industries, Des Moines, IA). The study consisted of three experimental periods (P). During P0 (2 d), all pigs were exposed to TN conditions (23 ± 3 °C, 68 ± 10% RH) and fed ad libitum. During P1 (7 d), all pigs received their respective dietary supplements, were maintained in TN conditions, and fed ad libitum. During P2 (21 d), HSAL, HSSO, and HSCr pigs were fed ad libitum and exposed to cyclical HS conditions (28 to 33 °C, 58 ± 10% RH). The TNAL and TNPF pigs remained in TN conditions and were fed ad libitum or pair-fed to their HSAL counterparts. Rectal temperature (TR), respiration rate (RR), and skin temperature (TS) were obtained daily at 0600 and 1800 h. At 1800 h, HS exposed pigs had increased TR, RR, and TS relative to TNAL controls (1.13 °C, 48 bpm, and 3.51 °C, respectively; P < 0.01). During wk 2 and 3 of P2, HSSO pigs had increased 1800 h TR relative to HSAL and HSCr (~0.40 and ~0.42 °C, respectively; P ≤ 0.05). Heat stress decreased ADFI and ADG compared to TNAL pigs (2.24 vs. 3.28 and 0.63 vs. 1.09 kg/d, respectively; P < 0.01) and neither variable was affected by SO or Cr supplementation. Heat stress increased or tended to increase moisture content of abdominal (7.7 vs. 5.9%; P = 0.07) and inner s.c. (11.4 vs. 9.8%; P < 0.05) adipose depots compared to TNAL controls. Interestingly, TNPF pigs also had increased adipose tissue moisture content and this was most pronounced in the outer s.c. depot (15.0 vs. 12.2%; P < 0.01) compared to TNAL pigs. Heat stress had little or no effect on fatty acid composition of abdominal, inner, and outer s.c. adipose tissue depots. In summary, the negative effects of HS on fat quality do not appear to be fatty acid composition related, but may be explained by increased adipose tissue moisture content.

Characterizing effects of feed restriction and glucagon-like peptide 2 administration on biomarkers of inflammation and intestinal morphology.

Inadequate feed consumption reduces intestinal barrier function in both ruminants and monogastrics. Objectives were to characterize how progressive feed restriction (FR) affects inflammation, metabolism, and intestinal morphology, and to investigate if glucagon-like peptide 2 (GLP2) administration influences the aforementioned responses. Twenty-eight Holstein cows (157 ± 9 d in milk) were enrolled in 2 experimental periods. Period 1 [5 d of ad libitum (AL) feed intake] served as baseline for period 2 (5 d), during which cows received 1 of 6 treatments: (1) 100% of AL feed intake (AL100; n = 3), (2) 80% of AL feed intake (n = 5), (3) 60% of AL feed intake (n = 5), (4) 40% of AL feed intake (AL40; n = 5), (5) 40% of AL feed intake + GLP2 administration (AL40G; 75 µg/kg of BW s.c. 2×/d; n = 5), or (6) 20% of AL feed intake (n = 5). As the magnitude of FR increased, body weight and milk yield decreased linearly. Blood urea nitrogen and insulin decreased, whereas nonesterified fatty acids and liver triglyceride content increased linearly with progressive FR. Circulating endotoxin, lipopolysaccharide binding protein, haptoglobin, serum amyloid A, and lymphocytes increased or tended to increase linearly with advancing FR. Circulating haptoglobin decreased (76%) and serum amyloid A tended to decrease (57%) in AL40G relative to AL40 cows. Cows in AL100, AL40, and AL40G treatments were euthanized to evaluate intestinal histology. Jejunum villus width, crypt depth, and goblet cell area, as well as ileum villus height, crypt depth, and goblet cell area, were reduced (36, 14, 52, 22, 28, and 25%, respectively) in AL40 cows compared with AL100 controls. Ileum cellular proliferation tended to be decreased (14%) in AL40 versus AL100 cows. Relative to AL40, AL40G cows had improved jejunum and ileum morphology, including increased villus height (46 and 51%), villus height to crypt depth ratio (38 and 35%), mucosal surface area (30 and 27%), cellular proliferation (43 and 36%), and goblet cell area (59 and 41%). Colon goblet cell area was also increased (48%) in AL40G relative to AL40 cows. In summary, progressive FR increased circulating markers of inflammation, which we speculate is due to increased intestinal permeability as demonstrated by changes in intestinal architecture. Furthermore, GLP2 improved intestinal morphology and ameliorated circulating markers of inflammation. Consequently, FR is a viable model to study consequences of intestinal barrier dysfunction and administering GLP2 appears to be an effective mitigation strategy to improve gut health.

Intentionally induced intestinal barrier dysfunction causes inflammation, affects metabolism, and reduces productivity in lactating Holstein cows.

Study objectives were to evaluate the effects of intentionally reduced intestinal barrier function on productivity, metabolism, and inflammatory indices in otherwise healthy dairy cows. Fourteen lactating Holstein cows (parity 2.6 ± 0.3; 117 ± 18 d in milk) were enrolled in 2 experimental periods. Period 1 (5 d) served as the baseline for period 2 (7 d), during which cows received 1 of 2 i.v. treatments twice per day: sterile saline or a gamma-secretase inhibitor (GSI; 1.5 mg/kg of body weight). Gamma-secretase inhibitors reduce intestinal barrier function by inhibiting crypt cell differentiation into absorptive enterocytes. During period 2, control cows receiving sterile saline were pair-fed (PF) to the GSI-treated cows, and all cows were killed at the end of period 2. Administering GSI increased goblet cell area 218, 70, and 28% in jejunum, ileum, and colon, respectively. In the jejunum, GSI-treated cows had increased crypt depth and reduced villus height, villus height-to-crypt depth ratio, cell proliferation, and mucosal surface area. Plasma lipopolysaccharide binding protein increased with time, and tended to be increased 42% in GSI-treated cows relative to PF controls on d 5 to 7. Circulating haptoglobin and serum amyloid A concentrations increased (585- and 4.4-fold, respectively) similarly in both treatments. Administering GSI progressively reduced dry matter intake (66%) and, by design, the pattern and magnitude of decreased nutrient intake was similar in PF controls. A similar progressive decrease (42%) in milk yield occurred in both treatments, but we observed no treatment effects on milk components. Cows treated with GSI tended to have increased plasma insulin (68%) and decreased circulating nonesterified fatty acids (29%) compared with PF cows. For both treatments, plasma glucose decreased with time while ß-hydroxybutyrate progressively increased. Liver triglycerides increased 221% from period 1 to sacrifice in both treatments. No differences were detected in liver weight, liver moisture, or body weight change. Intentionally compromising intestinal barrier function caused inflammation, altered metabolism, and markedly reduced feed intake and milk yield. Further, we demonstrated that progressive feed reduction appeared to cause leaky gut and inflammation.

Effects of dietary butyrate supplementation on intestinal integrity of heat-stressed cockerels.

Butyrate modulates intestinal epithelial cell structure and function. Three hundred and sixty Lohmann LSL-Classic layer cockerels were used to investigate the effect of butyrate on heat stress-induced intestinal injury and intestinal integrity. The experiment was conducted from day 85 to 105 of age. The birds were divided randomly into three treatments: control, heat stress (HS), and heat stress provided with butyrate (HSB) at a level of 0.35 g/kg of diet. The control was reared at 21 ± 1 °C throughout the experiment. The HS and HSB treatments were exposed to a cyclic HS (35 ± 1 °C from 09:00 to 13:00 and 21 ± 1 °C from 13:00 to 09:00). Intestinal and mucosal weights, villus height, villus surface area (VSA), absorptive epithelial cell area and intestinal beneficial bacteria were lower in the HS treatment than in the other two treatments (p < 0.05). Heat-stressed cockerels exhibited the highest (p < 0.05) villi injury scores and serum endotoxin levels compared with the other treatments. Dietary inclusion of butyrate increased (p < 0.05) intestinal and mucosal weights, villus height, VSA, absorptive epithelial cell area and intestinal beneficial bacteria counts and reduced (p < 0.05) HS-induced injury in intestinal epithelia as well as intestinal permeability to endotoxin. In conclusion, dietary butyrate exerted protective effects against intestinal damage induced by HS and improved intestinal health and integrity.

Glucose requirements of an activated immune system in lactating Holstein cows.

Accurately quantifying activated immune system energy requirements in vivo is difficult, but a better understanding may advance strategies to maximize animal productivity. Study objectives were to estimate whole-body glucose utilization following an i.v. endotoxin challenge. Lactating Holstein cows were jugular catheterized and assigned 1 of 3 bolus treatments: control (CON; 5 mL of saline; n = 6), lipopolysaccharide (LPS)-administered (LPS-C; 1.5 µg/kg of body weight; Escherichia coli 055:B5; n = 6), and LPS + euglycemic clamp (LPS-Eu; 1.5 µg/kg of body weight; 50% glucose solution infusion; n = 6). After LPS administration, blood glucose was determined every 10 min and glucose infusion rates were adjusted in LPS-Eu cows to maintain euglycemia for 720 min. Blood samples were obtained 180, 360, 540, and 720 min postbolus for further analysis. Cows were milked 360 and 720 min postbolus. Blood glucose was increased 84% in LPS-administered cows for up to 150 min postbolus; thereafter, circulating glucose was decreased 30% in LPS-C relative to LPS-Eu and CON cows. Mild hyperthermia (+0.5°C) occurred between 30 and 90 min postbolus in LPS-administered relative to CON cows; thereafter, rectal temperature did not differ between treatments. Milk yield and lactose percentage were decreased 80 and 11%, respectively, in LPS-administered relative to CON cows. Circulating insulin was increased 4 fold and nonesterified fatty acids, ß-hydroxybutyrate, and ionized Ca were decreased ∼50% in LPS-administered compared with CON cows. Plasma l-lactate, haptoglobin, and serum amyloid A increased ∼160, 260, and 75%, respectively, in LPS-administered relative to CON cows. Overall, LPS-binding protein was increased 87% in LPS-administered relative to CON cows; however, at 720 min, it was decreased 25% in LPS-Eu compared with LPS-C cows. White blood cell count decreased ∼90% in LPS-administered cows at 180 min and progressively increased to ∼50% of CON values by 720 min. Total glucose deficit during the 720 min following LPS administration was calculated as the decrease in the amount of glucose required to synthesize milk (due to the decrease in milk yield relative to prebolus levels) plus the amount of glucose infused to maintain euglycemia (in LPS-Eu cows only) and was 461, 1,259, and 1,553 g for CON, LPS-C, and LPS-Eu cows, respectively. Our data indicate an acutely activated immune system uses >1 kg of glucose within 720 min and maintaining euglycemia did not rescue milk synthesis.

Estimating glucose requirements of an activated immune system in growing pigs.

Activated immune cells become obligate glucose utilizers, and a large i.v. lipopolysaccharide (LPS) dose causes insulin resistance and severe hypoglycemia. Therefore, study objectives were to quantify the amount of glucose needed to maintain euglycemia following an endotoxin challenge as a proxy of leukocyte glucose requirements. Fifteen fasted crossbred gilts (30.3 ± 1.7 kg) were bilaterally jugular catheterized and assigned 1 of 2 i.v. bolus treatments: control (CON; 10 mL sterile saline; = 7) or LPS challenge + euglycemic clamp (LPS-Eu; 055:B5; 5 µg/kg BW; 50% dextrose infusion to maintain euglycemia; = 8). Following administration, blood glucose was determined every 10 min and dextrose infusion rates were adjusted in LPS-Eu pigs to maintain euglycemia for 8 h. Pigs were fasted for 8 h prior to the bolus and remained fasted throughout the challenge. Rectal temperature was increased in LPS-Eu pigs relative to CON pigs (39.8 vs. 38.8°C; < 0.01). Relative to the baseline, CON pigs had 20% decreased blood glucose from 300 to 480 min postbolus ( = 0.01) whereas circulating glucose content in LPS-Eu pigs did not differ ( = 0.96) from prebolus levels. A total of 116 ± 8 g of infused glucose was required to maintain euglycemia in LPS-Eu pigs. Relative to CON pigs, overall plasma insulin, blood urea nitrogen, ß-hydroxybutrate, lactate, and LPS-binding protein were increased in LPS-Eu pigs (295, 108, 29, 133, and 13%, respectively; ≤ 0.04) whereas NEFA was decreased (66%; < 0.01). Neutrophils in LPS-Eu pigs were decreased 84% at 120 min postbolus and returned to CON levels by 480 min ( < 0.01). Overall, lymphocytes, monocytes, eosinophils, and basophils were decreased in LPS-Eu pigs relative to CON pigs (75, 87, 70, and 50%, respectively; ≤ 0.05). These alterations in metabolism and the large amount of glucose needed to maintain euglycemia indicate nutrient repartitioning away from growth toward the immune system. Glucose is an important fuel for the immune system, and data from this study established that the glucose requirements of an intensely and acutely activated immune system in growing pigs are approximately 1.1 g/kg BW/h.

Technical note: A procedure to estimate glucose requirements of an activated immune system in steers.

Infection and inflammation impede efficient animal productivity. The activated immune system ostensibly requires large amounts of energy and nutrients otherwise destined for synthesis of agriculturally relevant products. Accurately determining the immune system's in vivo energy needs is difficult, but a better understanding may facilitate developing nutritional strategies to maximize productivity. The study objective was to estimate immune system glucose requirements following an i.v. lipopolysaccharide (LPS) challenge. Holstein steers (148 ± 9 kg; = 15) were jugular catheterized bilaterally and assigned to 1 of 3 i.v. TREATMENTS: control (CON; 3 mL saline; = 5), LPS-administered controls (LPS-C; 055:B5; 1.5 mg/kg BW; = 5), and LPS + euglycemic clamp (LPS-Eu; 1.5 mg/kg BW; 50% dextrose infusion to maintain euglycemia; = 5). In LPS-Eu steers, postbolus blood samples were analyzed for glucose every 10 min. Dextrose infusion rates were adjusted to maintain euglycemia for 720 min. All steers were fasted during the challenge. Samples for later analysis were obtained at 180, 360, 540, and 720 min relative to LPS administration. Rectal temperature was increased ∼0.5°C in both LPS treatments relative to CON steers ( = 0.01). Steers in both LPS treatments were hyperglycemic for ∼3 h postbolus; thereafter, blood glucose was markedly decreased (30%; < 0.01) in LPS-C relative to both CON and LPS-Eu steers. A total of 516 ± 65 g of infused glucose was required to maintain continuous euglycemia in LPS-Eu steers. Circulating insulin increased in LPS-C and LPS-Eu steers relative to CON (∼70% and ∼20 fold, respectively; < 0.01). Circulating NEFA increased similarly with time for both CON and LPS-C compared to LPS-Eu steers (∼43%; < 0.01). Plasma L-lactate and LPS binding protein increased (∼198 and ∼90%, respectively; < 0.01) and ionized calcium decreased (18%; < 0.01) in both LPS treatments relative to CON steers. Circulating white blood cells decreased initially in LPS-Eu and LPS-C relative to controls (180 min; 85%) followed by a progressive increase with time ( = 0.02). Blood neutrophils followed the same pattern; however, at 720 min, neutrophils were decreased in LPS-Eu compared to LPS-C, resulting in a decreased neutrophil-to-lymphocyte ratio (54%; = 0.03). The large amount of glucose needed to maintain euglycemia indicates extensive repartitioning of nutrients away from growth and the importance of glucose as a fuel for the immune system.