Milk microbiome in the first month of lactation and at weaning from ewes supplemented with zinc pre- and postpartum.

Mastitis is an important disease with economic and welfare implications in both clinical and subclinical states. The aim of this research was to sequence the hypervariable V4 region of the 16S rRNA gene to describe the microbial diversity and taxonomy of milk from clinically healthy ewes (Rambouillet, WF = 9; Hampshire, BF = 5). Experimental ewes represented a subset of a larger study assessing the impacts of divergent dietary zinc (Zn) concentrations [1× National Academics of Sciences, Engineering, and Medicine (NASEM) recommendations = CON or 3× NASEM recommendations = ZnTRT] throughout late gestation and lactation. Milk was collected at four periods during early lactation (18 - 24 h, 7 d, 14 d, and 21 d postpartum) and at weaning (84 ± 14 d postpartum). Somatic cell counts (SCC) were quantified, averaged, and classed (low: < 500 × 103; medium: 500 × 103 - 100 × 104; high: > 100 × 104 cells/mL). Milk samples (n = 67) were sequenced to identify bacteria and archaea; the most abundant phyla were Actinobacteria, Bacteroidetes, Cyanobacteria, Euryarchaeota, Firmicutes, Fusobacteria, Lentisphaerae, Proteobacteria, Spirochaetes, Tenericutes, Saccharibacteria TM7, and Verrucomicrobia. Mastitis pathogens were among the most relatively abundant genera, including Staphylococcus, Mannheimia, Corynebacterium, and Pseudomonas. Effects of breed, dietary Zn concentration, SCC class, and their two-way interactions on milk microbiome diversity and taxonomy were assessed within early lactation (using a repeated measures model) and weaning samples. Alpha-diversity metrics included Pielou's evenness, Faith's phylogenetic diversity, and Shannon's entropy indices. Main and interactive effects between Zn treatment, breed, SCC class, and period were variable in early lactation and not evident in weaning samples. Milk from BF ewes had increased Faith's phylogenetic diversity and Shannon's entropy, and differed in unweighted UniFrac composition (P ≤ 0.10). Milk from CON ewes had a reduced rate of composition change through early lactation (P = 0.02) indicating greater microbiome stability than ZnTRT ewe milk. These results support that milk is not sterile, and breed, dietary Zn concentration, and SCC class variably affect the milk microbiome. Findings from the current study provide important foundational insights into the effects of increased dietary Zn supplementation on longitudinal changes in the milk microbiome and associations with mammary gland health and mastitis.

Effects of management strategies during early lactation and weaning on etiological agents of ovine subclinical mastitis and antimicrobial susceptibility of milk-derived bacterial isolates.

Subclinical mastitis is a common intramammary disease in sheep production systems. Expenses associated with compromised animal performance, therapeutic interventions, and decreased ewe longevity make efforts to minimize its prevalence worthwhile. The objectives of this study were to 1) quantify the prevalence of subclinical mastitis throughout lactation, 2) evaluate the impact of bedding treatments on subclinical mastitis during early lactation, 3) evaluate the efficacy of prophylaxis and feed restriction during weaning on subclinical mastitis cure rates, and 4) identify levels and types of antimicrobial resistance in milk-derived bacteria. Ewe milk samples were collected at days 1, 2, and 28 post-partum, weaning, and 3-d post-weaning for bacterial identification via culture-based methods. Staphylococcus spp. and Streptococcus spp. isolates were subjected to in vitro antimicrobial susceptibility testing. The overall prevalence of subclinical mastitis defined by culture growth ranged between 22% and 66% and differences were observed between post-weaning and days 1 and 28 milk samples. Commonly isolated bacteria include coagulase-negative staphylococci (CoNS; 59%), Bacillus spp. (35%), Mannheimia haemolytica (10%), Staphylococcus aureus (8%), Streptococcus spp. (5%), and Corynebacterium spp. (5%). Early milk samples (days 1 and 2) were compared between jug bedding treatment: jugs were recently vacated, cleaned, and dusted with barn lime before adding fresh straw (CLEAN) or jugs were previously vacated and fresh straw was added atop soiled bedding (SOILED). Jug bedding treatment did not affect the prevalence of subclinical mastitis, though CoNS had greater sulfadimethoxine resistance in SOILED isolates than CLEAN isolates (P = 0.03). Three different weaning treatments were used: ewes were injected with penicillin at weaning (PENN), ewes had restricted feed access 48 h prior to and 72 h post-weaning (FAST), or a combination of these treatments (COMBO). Weaning treatment did not affect the prevalence of subclinical mastitis or cure rate from weaning to 3-d post-weaning, though all PENN and no FAST milk S. aureus isolates were resistant against tetracycline (P = 0.08). Subclinical mastitis prevalence tended to decrease from weaning to post-weaning (P = 0.08). These data show that subclinical mastitis is common throughout lactation and the levels of antimicrobial resistance of bacteria isolated from ewe milk are generally low against commonly used antimicrobials.

Subclinical mastitis is a common intramammary disease in livestock. Expenses associated with compromised animal performance, therapeutic interventions, and decreased ewe longevity make minimizing its prevalence worthwhile. The objectives of this study were to quantify the prevalence of subclinical mastitis, evaluate the impact of bedding treatments on subclinical mastitis, evaluate the efficacy of weaning treatments, and identify levels of antimicrobial resistance in milk-derived bacteria. The overall prevalence of subclinical mastitis was 45%. Common bacteria included coagulase-negative staphylococci (CoNS), Bacillus spp., Mannheimia haemolytica, Staphylococcus aureus, Corynebacterium spp., and Streptococcus spp. Early lactation milk samples were compared between jug bedding treatments: jugs were cleaned before adding fresh straw (CLEAN) or jugs had fresh straw added atop soiled bedding (SOILED). Jug bedding treatment did not affect the prevalence of subclinical mastitis, though did affect CoNS resistance to sulfadimethoxine. Three different weaning treatments were used: ewes were administered penicillin at weaning, ewes had restricted feed access at weaning, or a combination of the two treatments. Weaning treatment did not affect the prevalence of subclinical mastitis, though subclinical mastitis prevalence decreased post-weaning. Our data show that subclinical mastitis is generally prevalent throughout lactation, and the levels of antimicrobial resistance of bacteria isolated from ewe milk are generally low.

Relationships among intramammary health, udder and teat characteristics, and productivity of extensively managed ewes.

Mastitis is an economically important disease and its subclinical state is difficult to diagnose, which makes mitigation more challenging. The objectives of this study were to screen clinically healthy ewes in order to 1) identify cultivable microbial species in milk, 2) evaluate somatic cell count (SCC) thresholds associated with intramammary infection, and 3) estimate relationships between udder and teat morphometric traits, SCC, and ewe productivity. Milk was collected from two flocks in early (<5 d) and peak (30 to 45 d) lactation to quantify SCC (n = 530) and numerate cultivable microbial species by culture-based isolation followed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS; n = 243) identification. Within flock and lactation stage, 11% to 74% (mean = 36%) of samples were culture positive. More than 50 unique identifications were classified by MALDI-TOF MS analysis, and Bacillus licheniformis (18% to 27%), Micrococcus flavus (25%), Bacillus amyloliquefaciens (7% to 18%), and Staphylococcus epidermidis (26%) were among the most common within flock and across lactation stage. Optimum SCC thresholds to identify culture-positive samples ranged from 175 × 103 to 1,675 × 103 cells/mL. Ewe productivity was assessed as total 120-d adjusted litter weight (LW120) and analyzed within flock with breed, parity, year, and the linear covariate of log10 SCC (LSCC) at early or peak lactation. Although dependent on lactation stage and year, each 1-unit increase in LSCC (e.g., an increase in SCC from 100 × 103 to 1,000 × 103 cells/mL) was predicted to decrease LW120 between 9.5 and 16.1 kg when significant. Udder and teat traits included udder circumference, teat length, teat placement, and degree of separation of the udder halves. Correlations between traits were generally low to moderate within and across lactation stage and most were not consistently predictive of ewe LSCC. Overall, the frequencies of bacteria-positive milk samples indicated that subclinical mastitis (SCM) is common in these flocks and can impact ewe productivity. Therefore, future research is warranted to investigate pathways and timing of microbial invasion, genomic regions associated with susceptibility, and husbandry to mitigate the impact of SCM in extensively managed ewes.

Evaluation of Rambouillet, Polypay, and Romanov-White Dorper × Rambouillet ewes mated to terminal sires in an extensive rangeland production system: body weight and wool characteristics1,2.

Though lamb sales account for the majority of annual receipts on U.S. sheep operations, wool is an important income source for many Western flocks. Crossing fine-wool sheep with prolific or composite hair sheep breeds can increase lamb production, but fleece quality and marketability may be reduced by a greater content of nonwool fibers (e.g., med and kemp). The objectives of this study were to compare BW and wool characteristics of Rambouillet, Polypay, and Romanov-White Dorper × Rambouillet (RW-RA) ewes under extensive rangeland management conditions. Ewe BW was collected before mating (fall) and 30 d postlambing (spring) each year from 1 yr up to 4 yr of age. In spring and fall, Rambouillet and Polypay ewes were similar in BW (P ≥ 0.94). Spring BW did not differ (P = 0.13) between RW-RA and Polypay, but Rambouillet ewes were heavier than RW-RA in the spring (P = 0.02). Both Rambouillet and Polypay ewes were heavier (P < 0.07) than RW-RA in the fall. Greasy fleece weight and mid-side wool samples were collected from ewes at 1 and 4 yr of age. Clean fleece weights (CFW) were estimated from average laboratory scoured yield of mid-side wool samples composited within-breed. Average fiber diameter (AFD), SD of fiber diameter (SD-FD), and percentage med (%M), kemp (%K), and total medullated fibers (%T) were quantified on individual mid-side wool samples. There was no difference in 1-yr-old CFW among breed types (P ≥ 0.96). Four-yr-old Rambouillet ewes had heavier CFW (2.29 kg; P < 0.001) than 4-yr-old Polypay (1.83 kg) and RW-RA ewes (1.86 kg), which were not different (P > 0.99). Within 1- and 4-yr-olds, AFD differed among breed type (P < 0.001) and was the finest for Rambouillet (20.1 and 21.9 µm, respectively), intermediate for RW-RA (22.8 and 24.8 µm), and coarsest for Polypay (24.2 and 26.7 µm). Also within 1- and 4-yr olds, SD-FD was lowest in Rambouillet, intermediate in Polypay, and greatest in RW-RA (P < 0.01). Wool from RW-RA ewes had greater %M, %K, and %T (P < 0.001) than wool from Rambouillet and Polypay ewes, which were not different (P > 0.99). Results indicated superior wool production for Rambouillet compared with the coarser, more variable wool produced by Polypay and RW-RA. Still, past research reported greater lamb production in Polypay and RW-RA ewes which, under recent market conditions, would be associated with greater annual gross revenue for these breed types than for Rambouillet.

Effects of feeding juniper as a roughage on feedlot performance, carcass measurements, meat sensory attributes, and volatile aroma compounds of yearling Rambouillet wethers1,2.

The majority of U.S. lambs are born during late winter or early spring, which can create downstream variability in carcass quality if commercial lamb harvest is to be relatively constant throughout the year. Flavor is an important quality determining characteristic of sheep meat and is influenced, in part, by animal age at harvest. However, management practices to mitigate the risk of objectionable flavors in meat from old crop lambs or yearlings are not well known. Yearling (16.8 ± 0.14 mo) Rambouillet wethers were assigned to 1 of 3 treatment groups, which consisted of feeding a 20% ground sorghum-sudangrass hay diet for 40 d (JUN0; n = 10), a 20% ground juniper diet for 40 d (JUN40; n = 10), or a 20% ground hay diet for 20 d followed by a 20% ground juniper diet for 20 d (JUN20; n = 10). Wethers were harvested on day 41 and a whole bone-in loin and a boneless inside leg roast were fabricated from one side each of carcass. After grilling (loin chop) or convection air roasting (leg roast), trained sensory panel evaluation and measurement of aroma volatiles by gas chromatography/mass spectrometry were performed. Treatment diet did not affect (P ≥ 0.17) wether feedlot performance, dressing percentage, or loin eye area. However, wethers fed JUN0 tended (P = 0.06) to have greater back fat depth than wethers fed JUN20 or JUN40. No trained sensory panel trait of loin chop samples was affected (P > 0.10) by treatment. Leg roasts from JUN0 and JUN20 wethers had greater (P = 0.01) lamb identification sensory score than JUN40. Benzaldehyde, 1-heptanol, and 1-octanol concentrations were greater (P < 0.05) and decanal and nonenal concentrations were less (P < 0.05) in loin chops from JUN0 compared with JUN40 wethers. Additionally, the terpenes cedr-8-ene, gamma muurolene, and widdrene tended to be greater (P < 0.07) in loin chops from JUN20 and JUN40 than JUN0 wethers. The 2-pentyl-furan concentrations were greatest (P = 0.03) in leg roasts from JUN40 wethers. Like the loin chops, cedr-8-ene, gamma-muurolene, toluene, and widdrene were greater (P < 0.05) in leg roasts from wethers fed either of the juniper diets compared with JUN0. Yearling wethers can be finished on a feedlot diet containing 20% juniper for up to 40 d prior to harvest with no impact on feedlot performance, carcass characteristics, nor negative impact on sensory attributes or volatile compounds of either grilled loin chops or roasted legs.

Substituting ground woody plants for cottonseed hulls in kid goat feedlot diets: growth performance and blood serum chemistry.

Boer × Spanish kid goats (n = 48) were used to evaluate effects of using ground woody products in feedlot diets on growth performance and blood serum chemistry. A completely randomized study design was used with 2 feeding periods (Period 1 = 70% concentrate, days 0 to 26; Period 2 = 86% concentrate, days 27 to 64). Goats were individually fed 1 of 6 diets that differed only by roughage source (n = 4 wether males and 4 females/treatment; initial BW = 22 ± 2 kg): cottonseed hulls (CSH; control) or ground wood consisting of redberry (RED), blueberry (BLUE), one-seed (ONE), or eastern red cedar (ERC) Juniperus spp., or Prosopis glandulosa (MESQ). Ground woody diets were individually compared with CSH. During Period 1, goats fed CSH had greater (P < 0.05) average daily DMI (DMI), ADG, and G:F than goats fed MESQ and tended to have greater (P < 0.10) ADG and G:F than goats fed BLUE. A Treatment × d interaction (P = 0.008) was observed for goat BW during Period 1 and goats fed CSH tended (P < 0.09) to have greater BW on day 27 than goats fed BLUE or MESQ. During Period 2, Treatment × d interactions were not observed (P > 0.29) for DMI, ADG, G:F, or BW and no differences were observed between goats fed CSH and goats fed any of the treatment diets. Various blood serum variables were different between CSH and goats fed diets containing woody plants (mainly during Period 1); however, blood serum profiles did not indicate hepatotoxicity or any other health issue. Collectively, results suggested that ground Juniperus pinchotii, Juniperus ashei, or Juniperus monosperma can completely replace CSH in goat feedlot diets without negatively affecting growth performance or animal health. During Period 1, feeding diets to goats that contain 30% Juniperus virginiana (ERC) or P. glandulosa (MESQ) may not be economically justifiable in most scenarios, even though goat health, assessed by blood serum profiles, was not negatively affected. However, using 14% J. virginiana (ERC) or P. glandulosa (MESQ) in finishing diets is warranted.

Selenium supplementation restores innate and humoral immune responses in footrot-affected sheep.

Dietary selenium (Se) alters whole-blood Se concentrations in sheep, dependent upon Se source and dosage administered, but little is known about effects on immune function. We used footrot (FR) as a disease model to test the effects of supranutritional Se supplementation on immune function. To determine the effect of Se-source (organic Se-yeast, inorganic Na-selenite or Na-selenate) and Se-dosage (1, 3, 5 times FDA-permitted level) on FR severity, 120 ewes with and 120 ewes without FR were drenched weekly for 62 weeks with different Se sources and dosages (30 ewes/treatment group). Innate immunity was evaluated after 62 weeks of supplementation by measuring neutrophil bacterial killing ability. Adaptive immune function was evaluated by immunizing sheep with keyhole limpet hemocyanin (KLH). The antibody titer and delayed-type hypersensitivity skin test to KLH were used to assess humoral immunity and cell-mediated immunity, respectively. At baseline, FR-affected ewes had lower whole-blood and serum-Se concentrations; this difference was not observed after Se supplementation. Se supplementation increased neutrophil bacterial killing percentages in FR-affected sheep to percentages observed in supplemented and non-supplemented healthy sheep. Similarly, Se supplementation increased KLH antibody titers in FR-affected sheep to titers observed in healthy sheep. FR-affected sheep demonstrated suppressed cell-mediated immunity at 24 hours after intradermal KLH challenge, although there was no improvement with Se supplementation. We did not consistently prevent nor improve recovery from FR over the 62 week Se-treatment period. In conclusion, Se supplementation does not prevent FR, but does restore innate and humoral immune functions negatively affected by FR.

Effect of feeding selenium-fertilized alfalfa hay on performance of weaned beef calves.

Selenium (Se) is an essential micronutrient in cattle, and Se-deficiency can affect morbidity and mortality. Calves may have greater Se requirements during periods of stress, such as during the transitional period between weaning and movement to a feedlot. Previously, we showed that feeding Se-fertilized forage increases whole-blood (WB) Se concentrations in mature beef cows. Our current objective was to test whether feeding Se-fertilized forage increases WB-Se concentrations and performance in weaned beef calves. Recently weaned beef calves (n = 60) were blocked by body weight, randomly assigned to 4 groups, and fed an alfalfa hay based diet for 7 wk, which was harvested from fields fertilized with sodium-selenate at a rate of 0, 22.5, 45.0, or 89.9 g Se/ha. Blood samples were collected weekly and analyzed for WB-Se concentrations. Body weight and health status of calves were monitored during the 7-wk feeding trial. Increasing application rates of Se fertilizer resulted in increased alfalfa hay Se content for that cutting of alfalfa (0.07, 0.95, 1.55, 3.26 mg Se/kg dry matter for Se application rates of 0, 22.5, 45.0, or 89.9 g Se/ha, respectively). Feeding Se-fertilized alfalfa hay during the 7-wk preconditioning period increased WB-Se concentrations (P Linear<0.001) and body weights (P Linear = 0.002) depending upon the Se-application rate. Based upon our results we suggest that soil-Se fertilization is a potential management tool to improve Se-status and performance in weaned calves in areas with low soil-Se concentrations.