Efficacy of novel staphylococcal surface associated protein vaccines against Staphylococcus aureus and non-aureus staphylococcal mastitis in dairy cows.

Mastitis is an inflammation of the mammary gland commonly caused by bacteria or fungi. Staphylococcus aureus is a major bacterium that causes mastitis in dairy cows. Non-aureus staphylococci are also increasingly reported, with Staphylococcus chromogenes being the most common species. Current staphylococcal mastitis control programs are not fully effective, and treatment with antibiotics is not sustainable. Non-antibiotic sustainable control tools, such as effective vaccines, are critically needed. We previously developed S. aureus surface-associated proteins (SASP) and S. chromogenes surface-associated proteins (SCSP) vaccines that conferred partial protective effects. We hypothesized that vaccination with SASP or SCSP would reduce the incidence of S. aureus mastitis throughout the lactation period. The objective of this study was to evaluate the efficacy of SASP and SCSP vaccines against S. aureus and non-aureus staphylococcal mastitis under natural exposure over 300 days of lactation. Pregnant Holstein dairy cows (n = 45) were enrolled and assigned to receive SASP (n = 15) or SCSP (n = 16) vaccines or unvaccinated control (n = 14). Cows were vaccinated with 1.2 mg of SASP or SCSP with Emulsigen-D adjuvant. Control cows were injected with phosphate-buffered saline with Emulsigen-D adjuvant. Three vaccine injections were given subcutaneously at 60, 40, and 20 days before the expected calving. Booster vaccinations were given at 120 and 240 days in milk. Cows were monitored for mastitis at quarter and cow levels, staphylococcal mastitis incidence, changes in serum and milk anti-SASP and anti-SCSP antibody titers, bacterial counts in milk, adverse reactions, milk yield and milk somatic cells count over 300 days of lactation. The SCSP vaccine conferred a significant reduction in the incidence of staphylococcal mastitis. Milk and serum anti-SASP and anti-SCSP antibody titers were increased in the vaccinated cows compared to unvaccinated control cows. Anti-SASP and anti-SCSP antibody titers decreased at about 120 days in milk, indicating the duration of immunity of about four months. In conclusion, the SASP and SCSP vaccines conferred partial protection from natural infection.

The Impacts of Supplemental Protein during Development on Amino Acid Concentrations in the Uterus and Pregnancy Outcomes of Angus Heifers.

Replacement heifer development is one of the most critical components in beef production. The composition of the ideal uterine environment could maximize fertility and reproductive efficiency. Our hypothesis was that protein supplementation would affect the uterine environment of beef heifers without inhibiting development or reproduction. To test the effects of dietary supplementation on these outcomes, a randomized complete block design with repeated measures was implemented. Angus heifers (n = 60) were blocked by body weight (BW) and randomly assigned to one of three supplemental protein treatment groups (10% (CON), 20% (P20), and 40% (P40)). Mixed model ANOVAs were used to determine whether protein supplementation treatments, time, and the interaction or protein supplementation, semen exposure, and the interaction influenced uterine luminal fluid (ULF) and pregnancy outcomes. Amino acids (AAs) were impacted (p < 0.001), specifically, the essential AAs: Arg, Iso, Leu, Val, His, Lys, Met, Phe, Trp. Protein supplementation influenced multiple AAs post-insemination: Arg (p = 0.03), CC (p = 0.05), 1-MH (p = 0.001), and Orn (p = 0.03). In conclusion, protein supplementation did not affect the reproductive development via puberty attainment or the timing of conception even with alterations in growth. However, uterine AA concentrations did change throughout development and protein supplementation influenced ULF d 14 post-insemination, which may affect the conception rates.

Positive relationship of rectal temperature at fixed timed artificial insemination on pregnancy outcomes in beef cattle.

The overarching aim was to examine the relationship of rectal temperature at fixed time artificial insemination (FTAI) on pregnancy outcomes in a typical breeding season with expected pregnancy rates approaching 50% using Bos indicus and Bos taurus cattle. This represents a continuum of steps to test the hypothesis that elevated body temperature at or around insemination is functionally important to maximize pregnancy outcomes. Rectal temperature of Bos indicus cattle at FTAI ranged from 37.0 to 40.9 °C; 60.6% were hyperthermic. Positive factors impacting pregnancy outcomes were rectal temperature at FTAI, body condition, and estrus patch scores. Rectal temperature at FTAI was positively associated with pregnancy outcomes (P < 0.0001); per each 1 °C increase pregnancy odds increased 1.9 times (95% CI: 1.4 to 2.6). Highest pregnancy outcomes occurred with rectal temperatures exceeding 40 °C (P = 0.0004). Rectal temperature before FTAI in Bos taurus cattle ranged from 37.8 to 41.8 °C; 43.3% were hyperthermic. Factors impacting pregnancy were rectal temperature at FTAI, estrus activity, parity, and ambient conditions on day of FTAI. Rectal temperature of Bos taurus cattle at FTAI was positively associated with pregnancy (P = 0.0286); odds increased 1.45 times (95% CI: 1.0 to 2.0) per each 1 °C increase. Highest pregnancy outcomes occurred with rectal temperatures at FTAI exceeding 40 °C (P = 0.057). Moreover, positive relationship of rectal temperature at FTAI to pregnancy persisted in estrual females (71.25% of total; P = 0.0408; OR 1.5; 95% CI: 1.0 to 2.2). Mindful that 1) elevated temperatures observed in Bos indicus and Bos taurus cattle directly promote meiotic resumption of the oocyte in vitro and that 2) in vivo hyperthermia alters intrafollicular components which others have shown to potentiate ovulation and promote meiotic resumption, it is biologically plausible that an acute elevation in body temperature at or around time of insemination is functionally important to maximize pregnancy outcomes.

Reproductive efficiency remains a major challenge for beef producers with 35% to 55% of females failing to become pregnant after a single insemination. While basis for failure is multi-factorial, heightened estrus activity matters for pregnancy outcomes, even when synchronizing ovulation for fixed time artificial insemination. Body temperature increases of 1.5 °C+ are common during estrus. We hypothesize that higher estrous-associated temperatures (HEAT) at/near insemination are functionally important to maximize pregnancy outcomes. Elevated temperatures equivalent to what is observed in females exhibiting HEAT have been shown to induce oocyte meiotic resumption. An acute episode of hyperthermia after the LH surge alters intrafollicular components known to potentiate ovulation and affect the oocyte. Effort to examine the relationship of rectal temperature at fixed time artificial insemination with pregnancy outcomes in a breeding season with expected pregnancy rates >50% represents a next step in the continuum of hypothesis testing. The positive relationship of rectal temperature at insemination with pregnancy outcomes that was discovered adds to foundational knowledge. Because the degree of hyperthermia is related to highest pregnancy outcomes, a case is made for HEAT to be biologically and functionally important to maximize pregnancy outcomes in cattle.

A Surface Exposed, Two-Domain Lipoprotein Cargo of a Type XI Secretion System Promotes Colonization of Host Intestinal Epithelia Expressing Glycans.

The only known required component of the newly described Type XI secretion system (TXISS) is an outer membrane protein (OMP) of the DUF560 family. TXISSOMPs are broadly distributed across proteobacteria, but properties of the cargo proteins they secrete are largely unexplored. We report biophysical, histochemical, and phenotypic evidence that Xenorhabdus nematophila NilC is surface exposed. Biophysical data and structure predictions indicate that NilC is a two-domain protein with a C-terminal, 8-stranded ß-barrel. This structure has been noted as a common feature of TXISS effectors and may be important for interactions with the TXISSOMP. The NilC N-terminal domain is more enigmatic, but our results indicate it is ordered and forms a ß-sheet structure, and bioinformatics suggest structural similarities to carbohydrate-binding proteins. X. nematophila NilC and its presumptive TXISSOMP partner NilB are required for colonizing the anterior intestine of Steinernema carpocapsae nematodes: the receptacle of free-living, infective juveniles and the anterior intestinal cecum (AIC) in juveniles and adults. We show that, in adult nematodes, the AIC expresses a Wheat Germ Agglutinin (WGA)-reactive material, indicating the presence of N-acetylglucosamine or N-acetylneuraminic acid sugars on the AIC surface. A role for this material in colonization is supported by the fact that exogenous addition of WGA can inhibit AIC colonization by X. nematophila. Conversely, the addition of exogenous purified NilC increases the frequency with which X. nematophila is observed at the AIC, demonstrating that abundant extracellular NilC can enhance colonization. NilC may facilitate X. nematophila adherence to the nematode intestinal surface by binding to host glycans, it might support X. nematophila nutrition by cleaving sugars from the host surface, or it might help protect X. nematophila from nematode host immunity. Proteomic and metabolomic analyses of wild type X. nematophila compared to those lacking nilB and nilC revealed differences in cell wall and secreted polysaccharide metabolic pathways. Additionally, purified NilC is capable of binding peptidoglycan, suggesting that periplasmic NilC may interact with the bacterial cell wall. Overall, these findings support a model that NilB-regulated surface exposure of NilC mediates interactions between X. nematophila and host surface glycans during colonization. This is a previously unknown function for a TXISS.

Mapping and identification of QTL in 5601T × U99-310255 RIL population using SNP genotyping: soybean seed quality traits.

BACKGROUND: Molecular markers have played and will continue to play a major role in the genetic characterization and improvement of soybeans. They have helped identify major loci for tolerance to abiotic stressors, disease resistance, herbicide resistance, soybean seed quality traits, and yield. However, most yield quantitative trait loci (QTL) are specific to a certain population, and the genetic variation found in the specific bi-parental population is not always shared in other populations. A major objective in soybean breeding is to develop high yielding cultivars. Unfortunately, soybean seed yield, as well as protein and oil content, are complex quantitative traits to characterize from the phenotypic and genotypic perspectives. The objectives of this study are to detect soybean genomic regions that increase protein content, while maintaining oil content and seed yield and to successfully identify soybean QTL associated with these seed quality traits. METHODS AND RESULTS: To achieve these objectives, data from the 138 recombinant inbred lines grown in six environments were used to perform QTL detection analyses in search of significant genomic regions affecting soybean seed protein, oil, and yield. CONCLUSIONS: A total of 21 QTL were successfully identified for yield, protein, oil, methionine, threonine, lodging, maturity, and meal. Knowledge of their locations and flanking markers will aid in marker assisted selection for plant breeders. This will lead to a more valuable soybean for farmers, processors, and animal nutritionists.

The effects of differing nutritional levels and body condition score on scrotal circumference, motility, and morphology of bovine sperm.

Bulls often experience various levels of nutrient availability throughout the year. Nutritional management is a critical factor on overall ejaculate composition and the ability to get females pregnant. We hypothesized that differing nutritional levels and body condition score (BCS) affect reproductive fertility parameters in bulls. Mature Angus bulls (n = 11) were individually housed and randomly assigned to one of two dietary regimens: 1) over-fed (n = 5) or 2) restricted (n = 6). Bulls were fed the same ration at different volumes to achieve desired effects resulting in eight individual treatments: gain to an over-fed body condition score ([BCS]; GO), gain after nutrient restriction (GR), loss after an over-fed BCS (LO), loss from nutrient restriction (LR), maintenance at ideal adiposity (BCS = 6) after overfeeding (IMO), maintenance at ideal adiposity after nutrient restriction (IMR), maintenance at an over-fed BCS (BCS = 8; MO), and maintenance at a restricted BCS (BCS = 4; MR). Body weight (BW) and BCS were recorded every 2 wk to monitor bull weight and BCS changes. Scrotal circumference was measured every 28 d. Body fat and sperm motility and morphology were evaluated every 84 d. Scrotal circumference, motility, and morphology were normalized to the initial value of each bull. Thus, allowing the individual bull to serve as a control. Statistical analyses were conducted with PROC GLIMMIX of SAS as a complete randomized design to determine if treatment influenced BW, BCS, scrotal circumference, motility, morphology, and adipose thickness. Scrotal circumference (P < 0.001) had the least amount of deviation from initial during the LR (0.29 ±â€…0.44) treatment and the greatest during the MO (3.06 ±â€…0.44), LO (2.28 ±â€…0.44), MR (2.43 ±â€…0.44), GR (3.03 ±â€…0.44), and IMR (2.91 ±â€…0.44) treatments. Sperm motility was not affected by nutritional treatments (P = 0.55). Both head and total defects of sperm differed (P = 0.02) due to nutritional treatments. Increased head abnormalities occurred during the LO (37.60 ±â€…8.61) treatment, with no differences between the other treatments. Total defects increased during the LO (43.80 ±â€…9.55) treatment with similar increases in bulls during the GR (29.40 ±â€…9.55) and IMR (35.60 ±â€…9.55) treatments. In conclusion, male fertility was impacted when a deviation from a BCS of 6 occurred which could be detrimental to reproductive and beef production efficiency.

Hematological and immunological responses to naturally occurring bovine respiratory disease in newly received beef calves in a commercial stocker farm.

The objective was to determine temporal changes in hematological and immune parameters in response to naturally occurring bovine respiratory disease (BRD) in commercially managed stocker calves. Forty newly weaned beef steers purchased from auction markets were housed at a commercial stocker operation in Crossville, TN. Blood samples, rectal temperature, and thoracic ultrasonography (TUS; 1: normal to 3: severe) were collected on days 0, 7, 14, and 21. Castration status (FC: freshly castrated; PC: previously castrated) was determined on arrival based on presence of a fresh castration site at the scrotum. Calves received antibiotics for BRD based on clinical severity scoring (CSS; 0: moribund, 4: moribund) and rectal temperature. Complete blood counts (CBC) were performed. Calves were categorized based on the number of treatments (NumTrt) received (0x, 1x, and 2x). Temporal variations in CBC and immune parameters were analyzed using mixed model repeated measure ANOVA (Proc GLIMMIX; SAS 9.4). Variation of CBCs and immune parameters based on TUS was determined using mixed model ANOVA. There was a NumTrt by day interaction effect on the responses of white blood cells (WBC) (P = 0.04) and haptoglobin (HPT) (P = 0.04). On day 21, WBC were greater in the 2x NumTrt group than other groups, but there were no differences in WBC between NumTrt levels on other days. Haptoglobin was greater in the 2x group on days 14 and 21 than 0x or 1x. Red blood cells (RBC) (P = 0.02) and WBC (P = 0.04) differed between FC and PC groups, and lower RBC and WBC were observed in the FC group. A castration status × day effect for mean corpuscular volume (MCV; P = 0.04) was observed where FC group had higher MCV at days 14 and 21 than the PC group. Tumor necrosis factor-α differed based on NumTrt (P = 0.03) and higher concentrations were found in 2x group. We observed a day effect for IL-1ß (P = 0.009) and TNF-α (P = 0.001). Significant effect of TUS on HPT at day 14 (P = 0.0004) and day 21 (P = 0.002) was observed. Combining HPT and platelet explained 15% of the variability in treatment status on a given day, whereas HPT and hemoglobin explained 10% of the variability in lung consolidation status. Although hematological and immunological parameters varied largely in our study, the potential of combining HPT with hematological variables should be studied further. Results from this study would help in understanding temporal changes in CBC and immune parameters in newly received stocker cattle.

Blood and immune parameters are altered during bovine respiratory disease (BRD) progression and can be used for predicting disease status. We aimed looking at the dynamics of hematology and immunology in newly received stocker cattle in naturally occurring BRD. Forty newly received stocker cattle were managed by a local producer and monitored for BRD occurrence for 21 d after receiving during the high-risk period. Newly weaned calves were monitored as they experience several stress factors and become prone to BRD. Additionally, there are limited data related to immunological changes that occur in high-risk stocker cattle. Since there is no perfect diagnostic test for BRD, the diagnosis of BRD is likely missed when only visual signs are used. We observed that haptoglobin (HPT) was the most important parameter to differentiate BRD severity. The combination of HPT with blood parameters (hemoglobin and platelets) was useful to predict treatment and lung infection status. Therefore, measuring hematological and immunological parameters might be helpful to determine BRD status and facilitate treatment decisions in newly received stocker cattle.

Ruminal Protozoal Populations of Angus Steers Differing in Feed Efficiency.

Feed accounts for as much as 70% of beef production costs, and improvement of the efficiency with which animals convert feed to product has the potential to have substantial financial impact on the beef industry. The rumen microbiome plays a key role in determining feed efficiency; however, previous studies of rumen microbiota have not focused on protozoal communities despite the estimation that these organisms represent approximately 50% of rumen content biomass. Protozoal communities participate in the regulation of bacterial populations and nitrogen cycling-key aspects of microbiome dynamics. The present study focused on identifying potential associations of protozoal community profiles with feed efficiency. Weaned steers (n = 50) 7 months of age weighing approximately 260 kg were adapted to a growing ration and GrowSafe for 2 weeks prior to a 70-day feed efficiency trial. The GrowSafe system is a feeding system that monitors feed intake in real time. Body weights were collected on the first day and then every 7 days of the feed efficiency trial, and on the final day, approximately 50 mL of rumen content were collected via orogastric tubing and frozen at -80 °C. Body weight and feed intake were used to calculate residual feed intake (RFI) as a measure of feed efficiency, and steers were categorized as high (n = 14) or low (n = 10) RFI based on ±0.5 standard deviations about the mean RFI. Microbial DNA was extracted, and the eukaryotic component profiled by amplification and sequencing of 18S genes using degenerate primers that can amplify this locus across a range of protists. The taxonomy of protozoal sequences was assigned using QIIME 1.9 and analyzed using QIIME and SAS 9.4 with significance determined at α ≤ 0.05. Greater abundances of unassigned taxa were associated with high-RFI steers (p = 0.03), indicating a need for further study to identify component protozoal species. Differences were observed between low- and high-RFI steers in protozoal community phylogenetic diversity, including weighted beta-diversity (p = 0.04), Faith's phylogenetic diversity (p = 0.03), and observed Operational taxonomic unit (OTU) (p = 0.03). The unassigned taxa and differences in phylogenetic diversity of protozoal communities may contribute to divergences observed in feed efficiency phenotypes in beef steers.

The effects of feeding ferric citrate on ruminal bacteria, methanogenic archaea and methane production in growing beef steers.

Methane produced by cattle is one of the contributors of anthropogenic greenhouse gas. Methods to lessen methane emissions from cattle have been met with varying success; thus establishing consistent methods for decreasing methane production are imperative. Ferric iron may possibly act to decrease methane by acting as an alternative electron acceptor. The objective of this study was to assess the effect of ferric citrate on the rumen bacterial and archaeal communities and its impact on methane production. In this study, eight steers were used in a repeated Latin square design with 0, 250, 500 or 750 mg Fe/kg DM of ferric iron (as ferric citrate) in four different periods. Each period consisted of a 16 day adaptation period and 5 day sampling period. During each sampling period, methane production was measured, and rumen content was collected for bacterial and archaeal community analyses. Normally distributed data were analysed using a mixed model ANOVA using the GLIMMIX procedure of SAS, and non-normally distributed data were analysed in the same manner following ranking. Ferric citrate did not have any effect on bacterial community composition, methanogenic archaea nor methane production (P>0.05). Ferric citrate may not be a viable option to observe a ruminal response for decreases in enteric methane production.

Impact of an acute heat shock during in vitro maturation on interleukin 6 and its associated receptor component transcripts in bovine cumulus-oocyte complexes.

An acute heat stress event after the LH surge increased interleukin 6 (IL6) levels in the follicular fluid of the ovulatory follicle in hyperthermic cows. To examine direct consequences of a physiologically-relevant elevated temperature (41.0°C) on the cumulus-oocyte complex (COC), IL6 transcript abundance and related receptor components were evaluated throughout in vitro maturation. Heat-induced increases in IL6 were first noted at 4 hours of in vitro maturation (hIVM); peak levels occurred at 4.67 versus 6.44 hIVM for 41.0 and 38.5°C COCs, respectively (SEM = 0.23; P < 0.001). Peak IL6ST levels occurred at 6.95 versus 8.29 hIVM for 41.0 and 38.5°C, respectively (SEM = 0.23; P < 0.01). Transcript for LIF differed over time (P < 0.0001) but was not affected by 41.0°C exposure. Blastocyst development after performing IVF was not affected by 41.0°C exposure for 4 or 6 h. When limiting analysis to when IL6 was temporally produced, progesterone levels were only impacted by time and temperature (no interaction). Heat-induced shift in the temporal production of IL6 and IL6ST along with its impact on progesterone likely cooperate in heat-induced hastening of meiotic progression described by others.

Clinical Factors Associated With Survival Outcomes in Starved Equids: A Retrospective Case Series.

Each year in the United States, unwanted horses may become neglected, starved, or abandoned. Recovery therapies include refeeding and veterinary care, often requiring substantial time and financial investments. To better understand the likelihood for starved horses to successfully survive the first 100 days after initial evaluation, a retrospective case series was performed using hospital records of starved horses at the University of Tennessee College of Veterinary Medicine. A body condition score (BCS) of 3 or less and a malnourished diagnosis were utilized to select case records from an 11-year period (n = 82). Animal descriptors, vital signs, complete blood count data, blood chemistry panels, parasite load, and refeeding diet information were analyzed as available within the case record. Mixed model analyses of variance (PROC GLIMMIX, SAS 9.4) were utilized to test if response variables differed in horses that lived or died. Generalized linear mixed models were used to test factors associated with survival probability 100 days postinitial hospitalization. Body temperature (P = .05) and BCS (P = .0002) were significantly lower in horses that died compared with horses that survived. In addition, white blood cells were increased in horses that died (11.6 ± 1.3 cells/µL) compared with horses that survived (8.6 ± 0.5 cells/µL; P = .03). The initial BCS was associated with probability for survival, where survival likelihood increased 14.6 times for each 1 whole unit increase in the BCS. These results indicate the relevance of using the BCS as a predictor for future survival and as a measure for clinical decision-making.

The effects of protein level on cytokines and chemokines in the uterine environment of beef heifers during development.

The development of replacement heifers is crucial for breeding success and herd efficiency. Nutritional management can affect not only reproductive development but also the inflammatory status of the uterine environment, which may impact reproductive functions such as pregnancy establishment and development. The study herein evaluated the concentration of cytokines and chemokines in the uterus of heifers supplemented with different levels of protein. Angus heifers (n = 60) were blocked by body weight (BW) and randomly assigned to 1 of 3 treatments based on protein supplementation level: control of 10% crude protein (CON), 20% crude protein (P20), or 40% crude protein (P40). BW, body condition score, and blood samples were taken every 2 wk for 140 d to monitor development. Uterine flushes were performed monthly and concentrations of cytokines (IL-1α, IL-1ß, TNF-α, IFN-γ, IL-10, VEGF-α, IL-17A, and IL-36RA) and chemokines (IL-8, MCP-1, MIP-1α, and MIP-1ß) were quantified via ELISA multiplex. To test if there were mean differences in cytokines between the treatment groups or over time, PROC GLIMMIX (SAS v 9.4) was utilized. Concentrations of all cytokines and chemokines, except IL-1α, changed throughout heifer development (P < 0.05). Heifers in the P40 treatment group displayed reduced concentrations of MCP-1 (P = 0.007) and tended to have decreased concentrations of IFN-γ (P = 0.06). Cytokine IL-36RA tended (P = 0.06) to be affected by protein level, with the lowest concentrations observed in CON heifers. Most cytokines and chemokines increased following the initial month of supplementation (P < 0.05). The increase in concentrations after 1 mo may indicate an adaptive response in the uterus to diet change. Cytokines and chemokines fluctuated due to physiological changes occurring during development. Further research is needed to determine the influence of nutrition on uterine inflammation and long-term impacts on reproductive function.

Comparison of iron staining and scoring methods on canine bone marrow aspirates.

BACKGROUND: Insufficient iron for erythropoiesis can occur in multiple conditions, including absolute iron deficiency, which is often caused by chronic external hemorrhage in dogs. Distinguishing this from other causes of iron-restricted erythropoiesis allows appropriate intervention. Decreased marrow iron assessed by Prussian blue staining is a method to diagnose absolute iron deficiency, but scoring systems for marrow iron are not validated in dogs. OBJECTIVES: Our objectives were to (a) evaluate the technical performance of two bone marrow iron scoring systems used in human medicine and (b) examine the effects of destaining and restaining on iron stores after Wright-stained marrow slides are destained and restained with a Prussian blue stain. METHODS: Two marrow aspirate slides were included from each of 12 ill dogs in which marrow was collected during clinical evaluation. One slide was directly stained with Prussian blue. The other was first stained with Wright stain, then destained and restained with Prussian blue. Three blinded observers scored the presence of iron in each of the 24 randomized slides using the Gale (scale 0-6) and sideroblast methods (percentage score). Slides were then re-randomized and rescored. RESULTS: For the Gale method, interobserver agreement was fair, and intraobserver agreement was substantial to perfect. There was less agreement using the sideroblast method, with a significant observer effect. Iron scores were significantly lower in destained slides compared with those stained directly. CONCLUSIONS: Interobserver and intraobserver agreements were acceptable for the Gale method, but the sideroblast method should be used cautiously. A destaining procedure before Prussian blue staining could decrease marrow iron scores.

Soil Health Management Enhances Microbial Nitrogen Cycling Capacity and Activity.

Soil microbial transformations of nitrogen (N) can be affected by soil health management practices. Here, we report in situ seasonal dynamics of the population size (gene copy abundances) and functional activity (transcript copy abundances) of five bacterial genes involved in soil N cycling (ammonia-oxidizing bacteria [AOB] amoA, nifH, nirK, nirS, and nosZ) in a long-term continuous cotton production system under different management practices (cover crops, tillage, and inorganic N fertilization). Hairy vetch (Vicia villosa Roth), a leguminous cover crop, most effectively promoted the expression of N cycle genes, which persisted after cover crop termination throughout the growing season. Moreover, we observed similarly high or even higher N cycle gene transcript abundances under vetch with no fertilizer as no cover crop with N fertilization throughout the cover crop peak and cotton growing seasons (April, May, and October). Further, both the gene and transcript abundances of amoA and nosZ were positively correlated to soil nitrous oxide (N2O) emissions. We also found that the abundances of amoA genes and transcripts both positively correlated to field and incubated net nitrification rates. Together, our results revealed relationships between microbial functional capacity and activity and in situ soil N transformations under different agricultural seasons and soil management practices.IMPORTANCE Conservation agriculture practices that promote soil health have distinct and lasting effects on microbial populations involved with soil nitrogen (N) cycling. In particular, using a leguminous winter cover crop (hairy vetch) promoted the expression of key functional genes involved in soil N cycling, equaling or exceeding the effects of inorganic N fertilizer. Hairy vetch also left a legacy on soil nutrient capacity by promoting the continued activity of N cycling microbes after cover crop termination and into the main growing season. By examining both genes and transcripts involved in soil N cycling, we showed different responses of functional capacity (i.e., gene abundances) and functional activity (i.e., transcript abundances) to agricultural seasons and management practices, adding to our understanding of the effects of soil health management practices on microbial ecology.

Evaluation of the pharmacokinetics and efficacy of transdermal flunixin for pain mitigation following castration in goats.

The mitigation of pain associated with common management procedures is a rising concern among veterinarians, producers and consumers. Nonsteroidal anti-inflammatory drugs are vital compounds for this purpose due to their cost, convenience, and efficacy. A transdermal formulation of flunixin meglumine (FM) was approved for the treatment of pain in cattle; however, the efficacy has yet to be determined for small ruminants. The current study had two aims: 1) to determine the pharmacokinetics of transdermal flunixin meglumine (TD FM) in bucklings and 2) to evaluate pain mitigation by TD FM following castration. To evaluate pharmacokinetics, 12 male goats (mean age = 6 mo) received 2.2 mg/kg of FM IV (n = 6) or 3.3 mg/kg TD FM (n = 6). Plasma FM concentrations were measured. The mean C max, T max, and harmonic mean half-life for TD FM were 1.09 ± 0.65 µg/mL, 5.50 ± 2.95 h, and 7.16 ± 2.06 h, respectively. To evaluate the efficacy of pain mitigation, 18 goats were randomly assigned to three treatment groups: 1) TD FM and castration (FM CAST) (n = 6); 2) transdermal placebo and castration (PL CAST) (n = 6); and 3) TD FM and sham castration (SHAM) (n = 6). Plasma samples were collected at 0, 12, 24, 36, 48, 72, and 96 h to assess cortisol and prostaglandin E2 (PGE2). Daily dry matter intake (DMI) was recorded and body weight was measured at the beginning and end of the study. Thermography (IRT) images of the scrotum, as well as heart rate (HR), respiratory rate (RR), and rectal temperature, were taken twice daily. Separate mixed analysis of variance models were used to test the effects of treatment, time, and their interaction on mean body temperature, IRT, HR, and RR. Autoregressive covariance structure was utilized to account for repeated measures and individual goat DMI prior to the study was added as a covariate. There were no differences in vital parameters, IRT measurements, cortisol, or PGE2 in animals receiving either TD FM or placebo following castration (P > 0.05). DMI had a treatment by hour interaction and was significantly higher in FM CAST and SHAM groups than the PL CAST group (P = 0.04). Goats in the SHAM group gained weight throughout the study, whereas goats in all other groups lost weight (P = 0.02). Results indicate that TD FM may mitigate pain as demonstrated by increased DMI; however, a single dose may not be sufficient to reduce physiological indicators of pain associated with castration in goats.

Whole-Genome Sequencing and Bioinformatic Analysis of Environmental, Agricultural, and Human Campylobacter jejuni Isolates From East Tennessee.

As a leading cause of bacterial-derived gastroenteritis worldwide, Campylobacter jejuni has a significant impact on human health in both the developed and developing worlds. Despite its prevalence as a human pathogen, the source of these infections remains poorly understood due to the mutation frequency of the organism and past limitations of whole genome analysis. Recent advances in both whole genome sequencing and computational methods have allowed for the high-resolution analysis of intraspecies diversity, leading multiple groups to postulate that these approaches may be used to identify the sources of Campylobacter jejuni infection. To address this hypothesis, our group conducted a regionally and temporally restricted sampling of agricultural and environmental Campylobacter sources and compared isolated C. jejuni genomes to those that caused human infections in the same region during the same time period. Through a network analysis comparing genomes from various sources, we found that human C. jejuni isolates clustered with those isolated from cattle and chickens, indicating these as potential sources of human infection in the region.

Effect of stocking density and effective fiber on the ruminal bacterial communities in lactating Holstein cows.

Overstocking can be a major issue in the dairy cattle industry, leading to negative changes in feeding and resting behavior. Additional stress imposed and alterations in feeding behavior may significantly impact the rumen microbiome. The rumen microbiome is responsible for the successful conversion of feed to usable energy for its host. Thus, understanding the effects of stocking density on the rumen microbiome is imperative for further elucidation of potentially negative consequences of overstocking in dairy cattle. This study implemented a Latin Square design accounting for four pens of cattle and four treatment periods so that all treatment combinations were assigned to every pen during one period of the study. Two treatment factors, including two levels of physically effective neutral detergent fiber, achieved with addition of chopped straw, and stocking density (100% vs. 142%) of freestalls and headlocks, were combined and tested within a factorial treatment design. Within each pen, three or four cannulated cows (n = 15 total) were sampled for rumen content on the final day of each treatment period. Each treatment was randomly assigned to a single pen for a 14-day period. The V1-V3 hypervariable regions of the 16S rRNA gene were targeted for bacterial analyses. Variables with approximately normally-distributed residuals and a Shapiro-Wilk statistic of ≥0.85 were analyzed using a mixed model analysis of variance with the GLIMMIX procedure with fixed effects of feed (straw vs. no straw), stocking density (100% vs. 142%), and the interaction of feed × stocking density, and random effects of pen, period, feed × stocking × pen × period. Pen was included as the experimental unit in a given period and the sampling unit as cow. Variables included Shannon's Diversity Index, Faith's phylogenetic diversity index, chao1, observed OTU, and Simpson's evenness E as well as most individual taxa. Data were analyzed in SAS 9.4 utilizing the GLIMMIX procedure to perform mixed model analysis of variance. If data were not normally distributed, a ranked analysis was performed. No differences were observed in α-diversity metrics by fiber or stocking density (P > 0.05). Beta diversity was assessed using weighted and unweighted Unifrac distances in QIIME 1.9.1 and analyzed using ANOSIM. No differences were observed in weighted (P = 0.6660; R = -0.0121) nor unweighted (P = 0.9190; R = -0.0261) metrics and R values suggested similar bacterial communities among treatments. At the phylum level, Tenericutes differed among treatments with an interaction of stocking density by feed (P = 0.0066). At the genus level, several differences were observed by treatment, including Atopobium (P = 0.0129), unidentified members of order RF39 (P = 0.0139), and unidentified members of family Succinivibrionaceae (P = 0.0480). Although no diversity differences were observed, taxa differences may indicate that specific taxa are affected by the treatments, which may, in turn, affect animal production.

Dairy producers in the Southeast United States are concerned with cow care and welfare.

This research communication addresses the hypothesis that Southeast dairy producers' self-reported bulk tank somatic cell count (BTSCC) was associated with producers' response to three statements (1) 'a troublesome thing about mastitis is the worries it causes me,' (2) 'a troublesome thing about mastitis is that cows suffer,' and (3) 'my broad goals include taking good care of my cows and heifers.' Surveys were mailed to producers in Georgia, Kentucky, Mississippi, North Carolina, South Carolina, Tennessee, and Virginia (29% response rate, N = 596; final analysis N = 574), as part of a larger survey to assess Southeastern dairy producers' opinions related to BTSCC. Surveys contained 34 binomial (n = 9), Likert scale (n = 7), and descriptive (n = 18) statements targeted at producer self-assessment of herd records, management practices, and BTSCC. Statements 1 and 2 were assessed on a 5-point Likert scale from 'strongly disagree' to 'strongly agree.' Statement 3 was assessed on a 5-point Likert scale from 'very unimportant' to 'very important.' Reported mean BTSCC for all participants was 254 500 cells/ml. Separate univariable logistic regressions using generalized linear mixed models (SAS 9.4, Cary, NC, USA) with a random effect of farm, were performed to determine if BTSCC was associated with probability for a producer's response to statements. If BTSCC was significant, forward manual addition was performed until no additional variables were significant (P ≤ 0.05), but included BTSCC, regardless of significance. Bulk tank somatic cell count was associated with 'a troublesome thing about mastitis is the worries it causes me,' but not with Statements 2 or 3. This demonstrates that >75% of Southeastern dairy producers are concerned with animal care and cow suffering, regardless of BTSCC. Understanding Southeast producers' emphasis on cow care is necessary to create targeted management tools for herds with elevated BTSCC.

Prevalence of endoparasites in northern Mississippi shelter cats.

Parasitism of domestic cats impacts feline health and public health, when zoonotic parasites are present. Our objective was to evaluate endoparasite prevalence in cats from northern Mississippi animal shelters. Feline cadavers (n = 56) were collected from seven shelters from August 2017 to January 2018. Data included shelter, sex, reproductive status, intake date, originating source, and treatment records. Cadavers were processed to isolate stomach, and small and large intestines. Contents were strained and examined using stereomicroscopes for helminth collection and identification. Centrifugal flotation using Sheather's solution was performed on feces; urine sediments were also examined. Descriptive statistics in SAS was performed using the Frequency procedure. Kappa agreement statistics were obtained to determine agreement between fecal flotation and necropsy results. Separate logistic regression models were developed to test effects of risk factors on the probability for cats to test positive for outcomes of interest. Helminths were recovered in 82% of cats (46/56); specifically, Ancylostoma spp. (52%), Toxocara cati (43%), Taenia taeniaeformis (36%), Dipylidium caninum (29%), and Spirometra spp. (4%) were identified. Thirty-seven of 56 cats (66%) had parasite eggs or oocysts on fecal examination, including T. cati (39%), Ancylostoma spp. (34%), Cystoisospora spp. (23%), Spirometra spp. (9%), T. taeniaeformis (9%), and capillarid-type eggs (5%). Feline originating source was associated with presence of T. cati eggs in feces and presence of D. caninum in the gastrointestinal tract. Feral cats were more likely to have T. cati eggs in feces than owner surrender cats (OR 28; 95% CI: 1.9, 423), or stray cats (OR 8, 95% CI: 1.1, 57.0). Owner surrender cats were more likely to have D. caninum helminths in the gastrointestinal tract than stray cats (OR = 19.5; 95% CI: 2.0, 190). Toxocara cati exhibited strong agreement (κ = 0.70, 95% CI: 0.52, 0.89), Ancylostoma spp. exhibited moderate agreement (κ = 0.44, 95% CI: 0.22, 0.65), and cestodes exhibited poor agreement (κ = 0.02, 95% CI: -0.12, 0.15) between presence of eggs and gross helminths. Capillarid eggs (Pearsonema feliscati) were recovered in urine sediment of 6% (3/48) of cats. Overall, our study demonstrates a high level of parasitism in cats that entered Mississippi animal shelters. Parasites with zoonotic potential, such as Alaria spp., Ancylostoma spp., D. caninum, Physaloptera spp., T. taeniaeformis, T. cati, and Spirometra spp. were identified. Our results support the need for effective antiparasitic treatment of cats entering animal shelters in order to improve feline health and prevent environmental contamination with zoonotic parasites.

Effects of central and peripheral administration of an acute-phase protein, α-1-acid-glycoprotein, on feed intake and rectal temperature in sheep.

In rodents, an acute-phase protein, α-1-acid-glycoprotein (AGP), was shown to provide a link between inflammation and suppression of feed intake by acting as a leptin receptor agonist. The objective of this study was to determine the effects of AGP on feed intake and rectal temperature in sheep. Ewes were ovariectomized, implanted with a cannula into a lateral ventricle of the brain, and kept indoors in individual pens. Feed intake and rectal temperature were determined for sheep in all experiments. In the first experiment, ewes (n = 4) received 1 of 4 treatments [0 (control), 0.012 (low), 0.06 (medium), or 0.30 (high) mg/kg BW AGP] into the lateral ventricle (ICV). All sheep received all treatments in a Latin square design balanced for carryover effects with 10 d between treatments. In the second experiment, ewes (n = 10) received 1 of 2 treatments (0 and 3 mg/kg BW of AGP) intravenously (IV) in a completely randomized design. In the third experiment, ewes (n = 19) received peripheral treatments (IV) of an antipyretic [0 (control) or 2.2 mg/kg BW flunixin meglumine (FLU)] 30 min before receiving central AGP [0 (control) or 0.3 mg/kg BW of AGP] in a completely randomized design. All data were analyzed using a mixed model analysis of variance and tested for effects of treatment, time, and the interaction of treatment and time. Cumulative 48-h feed intake after administration of treatments was also determined. In the first experiment, there was no effect of ICV treatment (P = 0.37) on feed intake rate or on cumulative feed intake (P = 0.31). There was an effect of ICV treatment (P = 0.002) on rectal temperatures, which were greater (P < 0.05) after the high dose of centrally administered AGP. In the second experiment, there was no effect of AGP administration IV on feed intake rate (P = 0.98), on cumulative feed intake (P = 0.41) or on rectal temperature (P = 0.71). In the third experiment, there was an effect of central AGP treatment (P < 0.0001) and an interaction of central AGP and time (P < 0.0001) on rectal temperature, whereas FLU had no effect (P = 0.93), demonstrating that AGP increased rectal temperatures regardless of antipyretic treatment. These results indicate that central AGP increases rectal temperature in sheep by pathways that do not involve prostaglandins. Further research is needed to determine whether AGP may be an important integrator of energy balance and inflammation.