Croix Sheep Research Articles

Variants of NLRP3 Protein in Haemonchus contortus Infected Sheep: Impact on Immune Cell Responsiveness to LPS In Vitro

ABSTRACTPathogen recognition is an essential component to achieve the desired outcome of host protection. Nod‐like receptor pyrin containing domain 3 (NLRP3) is a cytoplasmic pattern recognition receptor (PRR) with a wide array of agonists, such as PAMPs, DAMPs, ATP, bacterial product and viral products. Stimulation of the NLRP3 inflammasome results in proteolytic activation of IL‐1β and IL‐18, cell pyroptosis and classically, the induction of proinflammatory responses. St. Croix (STC) sheep have resistance traits exhibiting the appropriate T‐helper type 2 immune response ensuing protection during helminth parasitic infection whereas parasite‐susceptible Suffolk (SUF) sheep have an impaired response resulting in parasite establishment and adverse symptoms. The objective of these experiments was to determine if NLRP3 protein in H. contortus‐infected SUF sheep was defective using the classical activation pathway of NLRP3 inflammasome. Peripheral blood mononuclear cells (PBMCs) derived from H. contortus‐infected STC and SUF sheep were isolated from whole blood and treated (MCC950 treatment for 2 h followed by LPS treatment for 3 h, 1400 W treatment for 2 h followed by LPS treatment for 3 h, LPS treatment for 3 h or culture media for 3 h). qPCR analysis of LPS‐stimulated PBMC revealed an upregulation in inflammatory associated genes IL‐1β, TLR4, TNFα and NFκB (p < 0.0001) in STC PBMC and downregulation in IFNγ, IL‐6 and iNOS for SUF PBMC. Pharmacological inhibition of iNOS in SUF PBMC resulted in an upregulation in the expression of IFNγ. These preliminary data begin to discover a relationship between NLRP3 activation and TLR4 signalling in PBMC of STC and SUF sheep.

Effect of a garlic drench on Galectin gene expression in ovine whole blood

Garlic, known for its immune-modulating and antibiotic properties, contains lectins that possess antimicrobial and immunomodulatory effects. Galectins (Gals), which bind β-galactosides, play a role in modulating immunity and pathological processes. It is hypothesized that garlic’s lectin components interfere with animal lectins. St. Croix sheep, known for their resistance to parasites and adaptability, are influenced by dietary supplements for innate immunity. This study evaluated the impact of garlic drench on Galectin gene expression in St. Croix sheep. Adult non-lactating ewes received either garlic juice concentrate or sterile distilled water for four weeks. Blood samples were collected, and plasma and whole blood cells were separated. Galectin secretion was assessed using a Sheep-specific ELISA, while Galectin gene transcription was analyzed through real-time PCR. Garlic administration upregulated LGALS-3 gene expression and significantly increased total plasma protein concentration. Garlic supplementation also affected Galectin secretion, with Gal-1, Gal-3, and Gal-9 showing differential effects.

21 Ancylostoma caninum-derived neutrophil inhibitory factor impairs ovine neutrophil chemotaxis to Haemonchus contortus larval antigen in Suffolk but not St. Croix sheep

Abstract Parasite-resistant St. Croix sheep (STC) generate a potent neutrophilic response to larval stages of Haemonchus contortus (Hc) as demonstrated by abomasal neutrophil infiltration as early as 3 days after infection. This phenomenon is delayed in parasite-susceptible Suffolk sheep (SUF) potentially contributing to larval establishment. An excretory/secretory (E/S) product, common to many helminths, is neutrophil inhibitory factor (NIF) which negatively affects neutrophil migration and activity. Due to differences in neutrophil accumulation between STC and SUF sheep, we hypothesized that Hc-NIF may inhibit migration of SUF and not STC neutrophils. To determine the concentration of NIF to use in subsequent experiments, neutrophils from STC and SUF were cultured with varying concentrations of Ancylostoma caninum-derived NIF (Ac-NIF) before exposure to larval Hc third-stage larval antigen (HcLA). Analysis of migration to stimuli revealed that the Ac-NIF concentration of 0.125 ug/mL yielded the greatest inhibition of migration towards HcLA between SUF and STC (8.70% and 11.64% migration, respectively; P = 0.0004).To test our hypothesis, neutrophils were cultured from STC and SUF sheep in the presence of Ancylostoma caninum-derived NIF (Ac-NIF) and measure chemotaxis to Interleukin-8 (IL-8), HcLA or Hc third-stage excretory/secretory (E/S) products. Neutrophils were isolated then incubated with Ac-NIF (0.125μg/mL) or complete media for 1 h. Neutrophils (1,000,000 cell/mL) were applied to cell migration inserts, and placed into a reservoir containing HcLA (20 μg/mL), E/S (20 μg/mL), IL-8 (50 ng/mL), or complete media. Migration plates were incubated (37°C, 5% CO2) for 24 h, after which, migrating cells were quantified using fluorescence. Ac-NIF inhibited SUF neutrophil migration towards IL-8 compared with STC (6.47% vs 9.95%, respectively; P = 0.0025). In response to HcLA alone, 64.0% of STC neutrophils migrated, while only 40.9% of SUF had the ability to migrate (P < 0.001). Towards HcLA, Ac-NIF inhibited SUF neutrophil migration compared with STC (8.40% vs 13.8%, respectively) (P < 0.001). Even more dramatically, in response to E/S products alone, only 24.3% of SUF neutrophils had the ability to migrate, while 76.4% of STC neutrophils migrated towards E/S (P < 0.001). In response to E/S, only 1.52% of SUF Ac-NIF-treated neutrophils were able to migrate, while 23.27% of STC neutrophils migrated (P < 0.001). Taken together, these data demonstrate that STC neutrophils are resistant to effects of neutrophil inhibition that is artificially applied or derived from HcE/S, which may explain breed differences in neutrophil migration in vivo.

PSV-26 Use of Garlic Drench in the Modulation of Galectin Gene Expression in St Croix Sheep

Abstract St. Croix sheep are parasite resistant and can live in a wide variety of climates. Dietary supplements can influence innate immunity. Garlic (Allium sativum) is used as an immune modulator and an alternative antibiotic. Lectins in garlic could interfere with animal lectins such as Galectins (Gals). Galectins bind β-galactosides and modulate pathological processes and immunity. The effect of garlic drench on Gal (1,3,9) gene expression in sheep blood was evaluated. All protocols were approved by the IACUC. Clinically healthy, adult St. Croix sheep were divided into two groups. The treatment group (n = 6) was drenched with 5 mL garlic barrier at a rate of 10 mL diluted in sterile distilled water 1:1. The control group (n = 6) was drenched with 10 mL sterile distilled water for four weeks. Cells and plasma were separated by centrifugation of whole blood. The concentration of total plasma protein was determined using the BCA assay kit (Pierce). Secretion of Galectins 1,3,9 was evaluated using Sheep-specific ELISA(My-BioSource). Data were analyzed using SAS Statistical analysis software 9.4 (SAS Institute). Nucleic acid purity and concentrations were assessed using the Nanodrop spectrophotometer. Galectin gene transcription was evaluated using sheep Gal 1,3 and 9 specific primers commercially sequenced (Eurofins Genomics). The house keeping genes, Ribosomal Protein Lateral Stalk Subunit P0 and Ubiquitin C-Terminal Hydrolase L5, were used as internal controls. Samples were amplified in duplicate using the CFX Connect real-time PCR system. The Livak Method was used to calculate the fold change in gene expression. Galectin 1, 3, and 9 were transcribed and translated. Transcription and translation were differentially affected by garlic. Galectin 1 transcription increased; secretion decreased. Galectin 3 transcription and secretion increased at P < 0.05. There was no change in the transcription of galectin 9, however, secretion was increased. Thus, garlic drench deferentially impacted transcription and secretion of structurally distinct Gal in sheep blood. Further studies are needed to determine the effect of Garlic on Gal-mediated functions important to sheep health and production.

PSII-3 Sheep Neutrophil Activation in Response to Garlic in Vitro

Abstract St. Croix hair sheep are known to resist parasitic infection. Parasitic resistance is in part due to the increased number and ability of neutrophil leukocytes to fight pathogens. Dietary supplements such as garlic (Allium sativum L.) are antimicrobial immune modulators that can enhance production. This study sought to evaluate the impact of fresh filtered garlic on sheep neutrophils through In vitro treatment. The study collected blood from the jugular vein of five clinically healthy female St. Croix sheep at NC A&T State University Farm and isolated neutrophils from 10 mL whole blood samples from each animal. The neutrophils were treated with fresh filtered garlic after the total cell count was measured, and 1 million cells were taken for treatment. The total protein concentration in the neutrophil supernatant was measured before and after treatment. Garlic (10 ug/mL PBS) was added to neutrophils, incubated (30 minutes, 37 °C) and centrifuged. Total RNA was extracted from the pellet using the TRIZOL RNA Isolation Protocol (Sigma-Aldrich). The RNA concentration and purity were assessed using a Nanodrop Spectrophotometer (Thermo Scientific Inc.). Pooled RNA from each group was used for cDNA synthesis, followed by qPCR analysis of the expression of 84 test genes related to the innate and adaptive immune response (RT2 profiler array) of the host. Total protein concentration in the supernatant was determined using the BCA Assay (Pierce BCA Protein Assay Kit). In vitro treatment with garlic extract resulted in increased total secreted protein concentrations from treated neutrophils, as well as upregulation of 10 genes involved in immune response and downregulation of 4 genes associated with inflammation and immune regulation. The study findings indicate that garlic stimulates gene transcription and protein secretion in neutrophils from St. Croix sheep in vitro. Studies are needed on the possible impacts on neutrophil function and animal response to infection for sustainable use of garlic in animal production.

121 Evaluation of Effects of Garlic Extract on Health Parameters and Gut Health in St. Croix Sheep

Abstract St. Croix sheep are hair sheep whose production characteristics make them suitable for low-input, sustainable production systems. They are parasite resistant and can live in a wide variety of climates. Natural immune modulators, such as garlic, can enhance animal production, health, and well-being. Garlic (Allium Sativa) is recognized as an eco-friendly alternative to antibiotics and contributes to a reduction in methane emissions. It has antibiotic, anti-helminthic and anti-inflammatory properties. The present study was carried out to determine the effect of garlic extracts on health variables and gut health indicators in St Croix sheep. Clinically, healthy adult, age-matched non-pregnant St Croix sheep (n = 12) from the NC A&T University farm were divided into treatment group (n = 6) and control group (n = 6). The treatment group was drenched with 5 mL garlic [Garlic Barrier (Glendale, CA)] concentrate diluted 1:1 with sterile distilled water (10 mL total). The control group was drenched with 10ml sterile distilled water for four weeks. The body weight, body condition, and FAMACHA scores and fecal samples were collected weekly. Fecal egg count was evaluated using the modified McMaster technique. Fecal samples were analyzed for eggs per gram using the McMaster slide. Total microbial DNA was isolated from fecal samples using Fast stool DNA Isolation kit (Qiagen, CA). DNA concentrations and purity were analyzed using a Nanodrop spectrophotometer. The relative abundance of Lactobacillus, bifidobacteria,16s RNA and Haemonchus are measured by using specific primers by running real time PCR. All variables were analyzed using the statistical analysis software package SAS version 9.4 (SAS Institute, Cary, NC). Garlic reduced fecal egg count significantly in weeks 1 (P < 0.034) and 3 (P < 0.003). There were no significant changes in fecal DNA Concentration. The relative of abundance of Lactobacillus is significantly increased whereas for Haemonchus decreased. Further studies are needed to define the pathways involved and consequences of changes in gut microbe community on immune function. These studies will aim in improved animal and human health and well-being and sustainable production.

158 Evaluating Lipopolysaccharide-Induced Behavioral and Immune Response Differences in Sheep Divergently Bred for Parasite Resistance

Abstract Katahdin lambs with decreased post weaning worm egg count estimated breeding values (PWEC EBV) generated greater antibody in response to vaccination and survive better to weaning. Thus, lambs with decreased PWEC values may be more disease resistant and immunocompetent. To test this hypothesis, parasite-resistant St. Croix sheep, (STC, n = 10), and parasite-susceptible Suffolk sheep (SUF, n = 10) were compared with Katahdin sheep divergent in PWEC EBV (HiKAT, n = 10; LoKAT, n = 10) and challenged with lipopolysaccharide (LPS). Sheep were intravenously administered LPS at a dose of 2.5 ug/kg; blood and body temperature data were collected hourly. Behavioral observations were recorded during sampling and videos within pens were obtained 30 min after blood collection. An increase of body temperature by 1 ºC was observed in all groups. Interestingly, STC and SUF temperature peaked at 4 hours whereas LoKAT and HiKAT both peaked at 6 hours. One metric of behavior recorded from video was Sheep Grimace Score (SGS; 1 = normal; 7 = extremely abnormal). Peak SGS in all groups was 3 hours after LPS administration. Main effects of group on SGS indicated a stepwise increase with STC having the least SGS (1.25) and SUF greatest score (2.45) and both groups of Katahdin sheep intermediate (1.65 LoKAT, 2.00 HiKAT; P < 0.001). Evaluation of “return to normal” (RTN), when SGS scores average between 0 and, revealed that STC and LoKAT RTN segregated from the other groups and this occurred at 5.1 and 6.6 hours after LPS treatment, respectively. HiKAT and SUF RTN was at 9.3 and 9.6 hours, respectively. Average SGS RTN in STC and LoKAT were significantly less than HiKAT and SUF (P < 0.01), indicating segregation in behavior. Typical behaviors associated with LPS response, such as fever and ill-behaviors, can be attributed to the rapid and systemic influx of proinflammatory cytokines like tumor necrosis factor alpha (TNFα). Analysis of circulating TNFα revealed similar segregation whereas STC and LoKAT have early TNFα peak at 4 hours and then gradually decline, HiKAT and SUF peak at 9hrs and remain elevated to2hrs. The transition period between these two disparate groups is 6 hours post LPS administration. Thus, TNFα was evaluated before (-2 to 6 hours) and post (7 to 12hours) TNFα shift. With STC sheep having greater TNFα before (45.96 pg/mL, P < 0.001) and SUF sheep having greater TNFα post (47.51 pg/mL, P < 0.001) indicating that despite similar average TNFα there exists timing differences of TNFα that correspond with SGS. These differences are mirrored in the production of TNFα in LoKAT and HiKAT sheep. Taken together these data indicate that early resolvement of TNFα cytokine production within the parasite-resistant groups (STC and LoKAT) is associated with reduced duration of sick behaviors.

162 The Effect of Innate Immune Receptor Activation on Differential Breed Responses toHaemonchus Contortus in Sheep

Abstract Previous studies have indicated the importance of Nod-like receptor pyrin containing domain 3 (NLRP3) to early immune response in St. Croix (STC) sheep. NLRP3 is important to Th2 immune response, which is the classical response to Haemonchus contortus larval antigen (HcLA). Th2 response is driven by interleukin (IL) -4 and IL-13 cytokine production and early production prevents parasite establishment and further resistance as seen in STC response to LPS, classical NLRP3 activation. Distinct differences have been seen in the structure of NLRP3 between both STC and SUF. STC upregulates inflammatory genes NLRP3, TLR4 and TLR2 in response to LPS, while SUF showed delayed responses leading to their susceptibility. The objective of this study was to investigate the role of NLRP3 in differential breed response, in the context of Hc infection. P Peripheral blood mononuclear cells (PBMC) from both SUF and STC were isolated and stimulated with HcLA antigen only, HcLA antigen in combination with pharmacological inhibitor of NLRP3 (MCC950) and complete media for 6 hours. Gene expression revealed that treatment with MCC950 upregulated TLR2 (P < 0.001), TLR4 (P < 0.001), STAT6 (P < 0.001), IL-4 and IL-10 (P < 0.001) in SUF PBMC while STC showed an increase in IL-1β (P < 0.001). This indicates that inhibition of NLRP3 improves response of SUF to Hc antigen and confirms importance of NLRP3 to immune response in STC.

122 Response of Ruminant Neutrophils on Fresh Filtered Garlic Preparation

Abstract The purpose of this study was to evaluate the effect of fresh filtered garlic preparation on ruminant neutrophils in-vitro. Blood was collected from the jugular vein of clinically healthy female St Croix sheep (n = 3) and Boer x Spanish goats (n = 3) at NC A&T State University Farm. Neutrophils were isolated from 10 mL whole blood sample of each animal. Fresh filtered garlic were prepared to stimulate the neutrophils. Before treating the neutrophils, the total cell count was measured, and 1 million cells were taken for treatment. Total protein concentration at different dilution levels of fresh filtered garlic were analyzed using BCA assay for the volumetric determination of garlic to be used at 10 ug/mL concentration. Before and after treatment the total protein concentration in the neutrophil supernatant were also measured. RNA was extracted from treated and untreated neutrophils pellet using TRIZOL RNA Isolation Protocol. The pooled RNA of goat and sheep samples were used for cDNA synthesis followed by qPCR analysis of galectin (Galectin-1, -3, -9) gene modulation in relation to indicators of animal health and production. Due to treatment, the total protein concentrations were increased. The magnitude of galectin gene expression was also observed in both sheep and goat neutrophils treated with fresh filtered garlic preparation. Galectin-3 and -9 expression increased manifolds in both sheep and goat neutrophils as compared with the control (untreated neutrophils) whereas galectin-1 increased in goat but decreased in sheep neutrophils. These results indicate that garlic had no adverse effect and differentially modulated galectin secretion in vitro. Galectins regulate biologic functions, immunity and disease pathogenesis and can serve as biomarkers. Further studies are warranted to determine the health and production benefits of the observed galectin modulation using garlic in small & large ruminants.

157 Effect of Sheep Breed on Neutrophil Chemotaxis toHaemonchus Contortus Larval Antigen

Abstract Parasite-resistant St. Croix sheep generate a potent neutrophilic response to larval stages of Haemonchus contortus. Protective neutrophil responses can be best characterized by increased neutrophil infiltration to the abomasum within the first 3 days of infection. These data indicate breed differences in neutrophil chemotaxis and response to H. contortus infection. The purpose of this study was to determine differences in neutrophil chemotaxis by H. contortus third-stage larval (L3) antigen (HcLA). Suffolk (SUF) and St. Croix (STC) neutrophils were isolated and applied to matrigel-coated clearview inserts, and placed into a reservoir containing H. contortus HcLA, Interleukin-8 (IL-8), or complete media. Neutrophil chemotaxis plates were incubated (37°C, 5% CO2) in an Incucyte S3 live cell imaging system for 24 hours, and measurements taken hourly. Neutrophils from STC migrated towards HcLA quicker than SUF neutrophils, and this migration occurred as early as 3 hours (10% and 4% respectively, P < 0.001). The difference in breed chemotaxis increased over time, where 28% of STC neutrophils migrated at 14 hours compared with 13% of SUF cells at the same time. By 24 hours, 37% of STC neutrophils migrated towards HcLA while the percentage of migrating SUF neutrophils remained the same (13%). Positive control for neutrophil chemotaxis was IL-8, and 37% of STC neutrophils achieved migration at 7 hours compared with 26% SUF neutrophils. By 24 hours, IL-8 induced neutrophil chemotaxis was similar in both breeds (STC 47%, SUF 41%). Taken together, these data indicate that delayed responses in neutrophil chemotaxis may contribute to H. contortus establishment in parasite-susceptible hosts. Thus, indicating that neutrophil response to H. contortus infection is a critical component to full host protective immunity.

Effects of Restricted Availability of Drinking Water on Blood Characteristics and Constituents in Dorper, Katahdin, and St. Croix Sheep from Different Regions of the USA.

Different hair sheep breeds originated from diverse climatic regions of the USA may show varying adaptability to water deprivation. Therefore, the objective of this study was to assess the effects of restricted availability of drinking water on blood characteristics and constituent concentrations in different breeds of hair sheep from various regions the USA. For this study, 45 Dorper (initial age = 3.7 ± 0.34 yr), 45 Katahdin (3.9 ± 0.36 yr), and 44 St. Croix (2.7 ± 0.29 yr) sheep from 45 farms in 4 regions of the USA (Midwest, Northwest, Southeast, and central Texas) were used. Ad libitum water intake was determined during wk 2 of period one, with 75% of ad libitum water intake offered during wk 2 of period two, and 50% of ad libitum water intake offered for 5 wk (i.e., wk 5−9) in period three. Water was offered at 07:00 or 07:30 h, with blood samples collected at 08:00 and(or) 14:00 h in wk 1, 2, 3, 4, 8, and 9 for variables such as hemoglobin and oxygen saturation and wk 2, 4, 6, 8, and 9 for concentrations of glucose and other constituents. The blood oxygen concentration at 08:00 h was 4.86, 4.93, and 5.25 mmol/L in period one and 4.89, 4.81, and 5.74 mmol/L in period three for Dorper, Katahdin, and St. Croix, respectively (SEM = 0.160; p = 0.001). Blood oxygen at 14:00 h was 4.37, 4.61, and 4.74 mmol/L in period one and 4.66, 4.81, and 5.46 mmol/L in period three for Dorper, Katahdin, and St. Croix, respectively (SEM = 0.154; p = 0.003). St. Croix were able to maintain a higher (p < 0.001) blood oxygen concentration than Dorper and Katahdin regardless of water availability. The pattern of change in blood concentrations with advancing time varied considerably among constituents. However, concentrations of glucose (55.3 and 56.2 mg/dL; SEM = 0.84), lactate (24.1 and 22.5 mg/dL; SEM = 0.79), total protein (7.08 and 7.17 g/dL; SEM = 0.0781), and albumin (2.59 and 2.65 g/dL in wk 2 and 9, respectively; SEM = 0.029) were similar (p > 0.05) between periods one and three. Conversely, concentrations of cholesterol (56.2 and 69.3 mg/dL; SEM = 1.33) and triglycerides (28.6 and 34.5 mg/dL in wk 2 and 9, respectively; SEM = 0.98) were greater (p < 0.05) in period three vs. 1. In conclusion, water restriction altered almost all the blood variables depending upon severity and duration of restriction, but the hair sheep breeds used from different regions of the USA, especially St. Croix, displayed considerable capacity to adapt to limited drinking water availability.

Efecto del inóculo (líquido ruminal o excreta) sobre la producción de gas in vitro en alimentos para ovinos

An experiment was carried out with the objective of evaluating the effect of two types of inocula from Saint Croix sheep (T1: ruminal fluid, T2: excreta with yeast), in two types of feed with different proportions of concentrate (C) and forage (F), (75C:25F; 25C:75F), on gas production at 24, 36 and 48 h of incubation (in duplicate) in 100 mL syringes (n = 3 replicates per diet), in order to determine in a factorial arrangement 2 X 2, (with two inocula and two diets), if the excreta can replace ruminal fluid as inoculum in in vitro gas production. Gas production was higher with ruminal fluid than with excreta, and with the high-concentrate diet. In conclusion, the excreta inoculum with yeast can be used to estimate the gas production obtained with ruminal fluid in in vitro incubations of feed for sheep.

PSVIII-9 Effect of Garlic Extracts on Galectin Protein Secretion in St. Croix Sheep Blood

Abstract The objective of this study was to evaluate the effect of garlic extract on the modulation of Galectin protein secretion in sheep blood. Dietary supplements can influence innate immunity. Garlic (Allium sativum) and its derivatives are eco-friendly non-antibiotic, growth-promoting dietary supplements known to modulate inflammation. Galectins (Gals) are a family of animal lectins that bind β-galactosides. Secreted Gal modulates pathological processes and inflammation. It is important to define potential targets of Garlic for its applications in sheep health. Clinically, healthy adult, age-matched non-pregnant St Croix sheep (n=12) from the NC A&amp;T University farm were divided into treatment group (n=6) and control group (n=6). Treatment group 1 was drenched with 5 ml garlic juice (Garlic Barrier (Glendale, CA) concentrate at a rate of 10 ml diluted in sterile distilled water 1:1. The control group was drenched with 10ml sterile distilled water for four weeks. The body weight, body condition, and FAMACHA scores were measured weekly. Faecal egg count was evaluated using the modified McMaster technique. Total plasma protein was analyzed using the BCA assay. Secretion of Gals 1, 3, 9 was evaluated using ELISA kits (My BIOSOURCE). Data were analyzed using T-test where p≤0.05 (SAS9.4 SAS Institute Cary NC). Garlic had no significant effect on FAMACHA ©, body condition score, packed cell volume, differential white blood cell count. Fecal egg count decreased significantly in treated animals in weeks 1(p&amp;lt; 0.034) and 3 (p&amp;lt; 0.003). Total protein, GAL-1 and GAL-3 concentration increased (p&amp;lt; 0.004) in the treatment group. The concentration of Gal-9 decreased when compared to the control group. Garlic had no adverse effect and differentially modulated Gal secretion. Further studies are required to determine the health and production benefits of the observed Gal modulation using garlic in St Croix sheep.

PSI-7 Differential Effects of Garlic on the Secretion of Structurally Different Galectins in Sheep Blood

Abstract The main objective of this study was to evaluate the effects of garlic treatment on structurally different Gals 1(Prototype), Gal 3 (chimera type), and Gal 9 (tandem-repeat type) secretion in sheep blood. St Croix sheep are parasite-resistant sheep useful for sustainable animal production. Producers seek to improve animal health and performance. Garlic (Allium sativum) and its derivatives are eco-friendly non-antibiotic growth-promoting dietary supplements. Garlic lectins have immune-modulatory effects that could interfere with animal lectins. Galectins (Gals) are a family of animal lectins that bind β-galactosides. Secreted Gals modulate pathological processes and immunity. A survey of (N=20) St Croix sheep was conducted to evaluate indicators of health and performance. Four clinically healthy female St Croix sheep (n=4) were used. Bodyweight (lb.), FAMACHA©, body condition score, blood, and fecal samples were collected. Blood was treated (37C for 30 min) with fresh Garlic (Herbal Roots CAs) at the following concentrations 0, 1, 10, 50,100 µg/ml PBS. Total plasma protein concentration was analyzed using BCA assay and secretion of Galectins 1,3 and 9 was determined using commercial ELISA(MY BIOSOURCE). All variables were analyzed using the statistical analysis software package SAS version 9.4(SAS Institute, Cary, NC) p≤0.05. Health indicators varied among sheep and no correlations were observed. The dose of garlic used and animal genetics had a variable effect. At 10ug/ml garlic increased concentration of total plasma protein (7%), Gal 1, 3(50% above control), and Gal-9 has no significant effect. concentration at 50 and 100 µg/ml garlic decreased all indicators. Galectin secretion was modulated by garlic treatment. There was a differential effect on the structurally distinct Gal types. Further studies are needed to determine the effect of Garlic on Gal mediated functions in St Croix sheep immunity and possible effects that can impact pathogenesis.

Effects of High Heat Load Conditions on Blood Constituent Concentrations in Dorper, Katahdin, and St. Croix Sheep from Different Regions of the USA.

Simple SummaryHigh heat load (HL) can adversely affect livestock health and cause significant production losses as a result of alterations of many physiological functions including nutrient metabolism, oxidative status, blood constituent levels, and gut dysbiosis. Effects of high HL could vary with breed of hair sheep (i.e., Dorper, Katahdin, and St. Croix) as well as region of the United States of America with different climatic conditions where they originated. Differences among the three sheep breeds in levels of a number of blood constituents were observed. Different HL conditions also altered concentrations of several blood metabolites and constituents, including hemoglobin, oxygen concentration, creatinine, heat shock protein, thyroxine, and cortisol. However, there were no interactions between breed and HL for most blood variables, suggesting similar resilience to high HL. In addition, only a few differences among or interactions involving region suggest that the breeds are highly adapted to diverse climatic conditions.Forty-six Dorper (DOR), 46 Katahdin (KAT), and 43 St. Croix (STC) female sheep (initial body weight of 58, 59, and 46 kg, respectively, SEM = 1.75; 3.3 ± 0.18 years of age, 2.6–3.7), derived from 45 commercial farms in four regions of the USA (Midwest, Northwest, Southeast, and central Texas), were used to evaluate responses in blood constituent concentrations to increasing heat load index (HLI) conditions. There were four sequential 2 weeks periods with target HLI during day/nighttime of 70/70 (thermoneutral zone conditions), 85/70, 90/77, and 95/81 in period 1, 2, 3 and 4, respectively. A 50% concentrate pelletized diet was fed at 53.3 g dry matter/kg body weight0.75. The analysis of most constituents was for samples collected on the last day of the second week of each period at 13:00 h; samples for cortisol, thyroxine, and heat shock protein were collected in week 2 and 8. Previously, it was noted that resilience to high HLI conditions was greatest for STC, lowest for DOR, and intermediate for KAT. There were few effects of region. Other than hemoglobin concentration, there were no interactions between breed and period. Blood oxygen concentration was greatest (p < 0.05) among breeds for STC (5.07, 5.20, and 5.53 mmol/L for DOR, KAT, and STC, respectively; SEM = 0.114) and differed among periods (4.92, 5.26, 5.36, and 5.52 mmol/L for period 1, 2, 3, and 4, respectively; SEM = 0.093). There were breed differences (i.e., main effects; p < 0.05) in glucose (50.0, 52.6, and 52.1 mg/dL; SEM = 0.76), urea nitrogen (17.2, 17.3, and 19.4 mg/dL; SEM = 0.33), creatinine (0.991, 0.862, and 0.802 mg/dL; SEM = 0.0151), total protein (6.50, 6.68, and 6.95 g/l; SEM = 0.017), triglycerides (28.4, 29.1, and 23.5 mg/dL; SEM = 0.87), and cortisol (6.30, 8.79, and 6.22 ng/mL for DOR, KAT, and STC, respectively; SEM = 0.596). Differences among periods (p < 0.05) were observed for lactate (27.9, 25.3, 27.8, and 24.0 mg/dL; SEM = 0.99), creatinine (0.839, 0.913, 0.871, and 0.917 mg/dL; SEM = 0.0128), total protein (6.94, 6.66, 6.60, and 6.65 g/l; SEM = 0.094), and cholesterol (60.2, 56.5, 58.3, and 57.6 mg/dL for period 1, 2, 3, and 4, respectively; SEM = 1.26). In addition, the concentration of cortisol (7.62 and 6.59 ng/mL; SEM = 0.404), thyroxine (5.83 and 5.00 µg/dL; SEM = 0.140), and heat shock protein (136 and 146 ng/mL for week 2 and 8, respectively; SEM = 4.0) differed between weeks (p < 0.05). In conclusion, the lack of interaction between breed and period with different HLI conditions suggests that levels of these blood constituents were not highly related to resilience to high HLI.

Management of St. Croix Sheep and Garut Sheep Genetic Resources in Sheep Formation

This study aimed to determine the management of the genetic resources of St. Croix sheep and Garut sheep in the formation of sheep clumps and their physiological conditions. The study was conducted at the Livestock Research Institute on Jalan Raya Pajajaran Bogor, West Java, in 2019. Ex-situ collection materials and characterization of St. Croix sheep and Garut sheep were as many as 250 heads; sheep breeding was carried out in groups; one male married 5-10 heads. The rams were left in the mating group for two oestrus cycles (34 days). Primary data and secondary data were analyzed descriptively and quantitatively. The results were obtained in the germplasm research of St. Croix with an average mating body weight of 23.45 ± 4.08 kg and Garut sheep of 26.20 ± 4.92 kg. The average birth weight of St. Croix sheep was 2.26+0.35 kg, and Garut sheep was 2.05± 0.47kg. The establishment of the St. Croix and Garut sheep have high genetic quality, production, and reproductive characteristics, but each has a different productivity level. The study of germplasm preservation of sheep was carried out to increase the population of St. Croix sheep and pure Garut sheep so that the maximum target population of &gt;200 sheep was achieved to meet the demand for seeds as animal food.

PSXIV-18 Genetic improvement for resistance to gastrointestinal parasitism in growing meat goats and hair sheep through a centralized performance test in the southcentral USA

Abstract Progress from selecting meat goats and hair sheep from different farms in the southcentral United States for resistance to internal parasitism during three consecutive central performance tests was evaluated. Tests entailed 7–10 wk of data collection after artificial infection of growing males with Haemonchus contortus larvae. There were 44, 50, and 50 Boer, Kiko, and Spanish goats and 54, 55, 34, and 47 Dorper, Katahdin-farm-A, Katahdin-farm-B, and St. Croix sheep, respectively. In year 1, males were randomly selected and in years 2 and 3 were progeny of sires and females classified as highly and moderately resistant. Criteria for males was average daily gain and residual feed intake during the performance test and means of fecal egg count (FEC) and packed cell volume after artificial infection and for females was FEC and FAMACHA on-farm. There were interactions in FEC of breed×year (P &amp;lt; 0.001) for both species (1,046; 1,134; and 2,630 for Boer; 3,781; 1,239; and 1,216 for Kiko; 1,303; 1,357; and 1,464 for Spanish; 3,288; 1,956; and 1,315 for Dorper; 1,528; 1,759; and 2,241 for Katahdin-A, 2,414; 3,874; and 1,793 for Katahdin-B; and 1,371; 1,160; and 1,175 eggs/g for St. Croix in years 1, 2, and 3, respectively) and year×resistance group (RG; P = 0.005) for goats (1,359; 610, and 552 for high; 1,951; 1,268; and 1,440 for moderate; 2,821; 1,852; and 3,118 eggs/g for low RG in years 1, 2, and 3, respectively) but not for sheep (P = 0.639). In conclusion, resistance appeared greatest for Spanish and St. Croix within species and was increased by selection of Kiko and Dorper. For highly resistant goats, lower FEC in year 2 and 3 vs. 1, compared with no year differences for moderate and low RG, indicates selection progress, and the lack of similar differences for sheep may reflect their relatively greater resistance.

PSXII-29 Evaluation of Barbados Blackbelly and St. Croix sheep under accelerated lambing using purebred and terminal sire mating: Lamb survival and body weight

Abstract Objectives were to evaluate lamb survival and body weight (BW) under an accelerated, pasture-lambing system. Lambs were born to Barbados Blackbelly (BB) or St. Croix dams (SC) mated within breed (purebred) or to Dorset (terminal) in single sire mating groups. Lambs were born in April (2013 and 2015), December (2013 and 2015), and August (2014 and 2016) and weaned at 9 weeks of age. Traits considered were BW at birth (BW0; n = 959), 65-d adjusted weaning BW (BW65; n = 788), and survival to weaning (n = 959). Data were analyzed with fixed effects of dam age, sex, birth type, dam breed (BB vs SC), dam mating system (purebred vs terminal), birth month-year, and significant two-way interactions and a random dam effect. Dam breed did not impact BW65 (P = 0.09). While BW0 was greater for lambs born to SC than BB dams (3.44 vs 3.23 kg; P &amp;lt; 0.01), survival to weaning was greater for lambs born to BB than SC dams (0.93 vs. 0.87; P = 0.02). The dam mating system x birth month-year interaction was significant for both BW traits (P ≤ 0.01). Within birth month-year, BW0 was 18 to 24% greater and BW65 was 12 to 24% greater for terminal than purebred lambs (P &amp;lt; 0.01). However, BW0 differences between dam mating systems were greatest for August born lambs whereas BW65 differences were greatest for April born lambs. Lamb survival was not affected by dam mating system or birth month-year (P ≥ 0.27). Using a terminal sire improved BW without affecting survival of lambs reared by landrace hair sheep dams, making this a viable management tool to increase productivity of forage-based production systems.

Effects of restricted feed intake on blood constituent concentrations in Dorper, Katahdin, and St. Croix sheep from different regions of the USA

Forty-six Dorper, 47 Katahdin, and 41 St. Croix female sheep (initial BW = 62, 62, and 51 kg, respectively, SEM = 1.43; 3.8 ± 0.18 yr) from farms in Midwest, Northwest, Southeast, and central Texas regions of the USA were used to evaluate effects of feed restriction on blood constituent levels. The amount of feed offered varied in the first 4 wk to achieve stable BW, and that in wk 5–10 was 55% of intake in wk 3–4. Blood was sampled at the end of wk 3, 4, 6, 8, and 10. There were relatively few effects and interactions involving region and no breed × time interactions. Breed affected the concentration of a small number of constituents, including urea N (14.0, 13.7, and 15.4 mg/dl; SEM = 0.31) and creatinine (0.945, 0.836, and 0.809 mg/dl for Dorper, Katahdin, and St. Croix, respectively; SEM = 0.0253). Also, the concentration of triglycerides in wk 4 and 10 was lowest for St. Croix (29.8, 29.5, and 26.7 mg/dl for Dorper, Katahdin, and St. Croix, respectively; SEM = 0.88). There was a trend for a difference (P = 0.051) between wk 4 and 10 in the glucose concentration (51.9 and 54.2 mg/dl; SEM = 0.90), and there were differences (P < 0.05) in lactate (23.9 and 20.3 mg/dl; SEM = 0.89), urea N (16.4 and 13.0 mg/dl; SEM = 0.25), creatinine (0.808 and 0.919 mg/dl; SEM = 0.0165), triglycerides (31.8 and 25.5 mg/dl; SEM = 0.63), and cholesterol (67.5 and 74.7 mg/dl, respectively; SEM = 1.66). In conclusion, similar responses in blood constituent levels of different hair sheep breeds to feed restriction is in accordance with comparable effects on the maintenance energy requirement.

PSXII-30 Genetic and phenotypic correlations for growth and resistance to gastrointestinal parasitism in meat goats and hair sheep

Abstract Genetic and phenotypic correlations for growth and response to parasite infection traits were estimated for growing male meat goats and hair sheep from different farms in the southcentral United States during three consecutive central performance tests (CPT). Data were collected for 7–10 wk after artificial infection with Haemonchus contortus, which included average daily gain (ADG), fecal egg count (FEC), packed cell volume (PCV), and immunoglobin (Ig) levels. Animals evaluated were selected randomly in year 1 and in years 2 and 3 progeny of CPT sires classified as highly or moderately resistant and included 46, 50, and 51 Boer, Kiko, and Spanish goats and 59, 61, 34, and 46 Dorper, Katahdin-farm A, Katahdin-farm B, and St. Croix sheep, respectively. Females were classified accordingly on-farm based on FEC and FAMACHA. Pedigree records included 4 and 10 full-sibs and 97 and 149 half-sibs for goats and sheep, respectively. Variance components and correlations were estimated by AIREML using WOMBAT with a multivariate animal model. The additive genetic correlation between FEC and PCV was negative for goats (r=-041, P &amp;lt; 0.001) but positive for sheep (r=0.21, P = 0.004), whereas the phenotypic correlation between FEC and PCV was nonsignificant for goats but negative for sheep (r=-0.252, P &amp;lt; 0.001). The genetic correlation between FEC and IgA was positive (r=0.39, P &amp;lt; 0.001) for goats but nonsignificant (P = 0.439) for sheep, whereas those of FEC with IgM and IgG were both negative (r=-0.369 and -0.732 with IgM and r=-0.284 and -0.702 for goats and sheep, respectively; P &amp;lt; 0.001). Genetic and phenotypic correlations between ADG and FEC were nonsignificant for both species. In conclusion, different relationships of FEC and PCV between species require careful attention during selection and the lack of relationship between ADG and FEC suggests that selection of growing male meat goats and hair sheep for resistance to internal parasitism will not adversely affect growth performance.