Effect of bone and analytical method on assessment of bone mineralization in response to dietary phosphorus, phytase, and vitamin D in finishing pigs.

A total of 882 pigs (PIC TR4 × [Fast LW × PIC L02]; initially 33.2 ±â€…0.31 kg) were used in a 112-d study to evaluate the effects of different bones and analytical methods on the assessment of bone mineralization response to changes in dietary P, phytase, and vitamin D in growing pigs. Pens of pigs (20 pigs per pen) were randomized to one of five dietary treatments with nine pens per treatment. Dietary treatments were designed to create differences in bone mineralization and included: 1) P at 80% of NRC (2012) standardized total tract digestible (STTD) P requirement, 2) NRC STTD P with no phytase, 3) NRC STTD P with phytase providing an assumed release of 0.14% STTD P from 2,000 FYT/kg, 4) high STTD P (128% of the NRC P) using monocalcium phosphate and phytase, and 5) diet 4 with additional vitamin D3 from 25(OH)D3. On day 112, one pig per pen was euthanized for bone, blood, and urine analysis. Additionally, 11 pigs identified as having poor body condition which indicated a history of low feed intake (unhealthy) were sampled. There were no differences between treatments for final body weight, average daily gain, average daily feed intake, gain to feed, or bone ash measurements (treatment × bone interaction) regardless of bone ash method. The response to treatment for bone density and bone mineral content was dependent upon the bone sampled (density interaction, P = 0.053; mineral interaction, P = 0.078). For 10th rib bone density, pigs fed high levels of P had increased (P < 0.05) bone density compared with pigs fed NRC levels with phytase, with pigs fed deficient P, NRC levels of P with no phytase, and high STTD P with extra 25(OH)D3 intermediate, with no differences for metacarpals, fibulas, or 2nd ribs. Pigs fed extra vitamin D from 25(OH)D3 had increased (P < 0.05) 10th rib bone mineral content compared with pigs fed deficient P and NRC levels of P with phytase, with pigs fed industry P and vitamin D, and NRC P with monocalcium intermediate. Healthy pigs had greater (P < 0.05) serum Ca, P, vitamin D concentrations, and defatted bone ash than those unhealthy, with no difference between the two health statuses for non-defatted bone ash. In summary, differences between bone ash procedures were more apparent than differences between diets. Differences in bone density and mineral content in response to dietary P and vitamin D were most apparent with 10th ribs.

Lameness is defined as impaired movement or deviation from normal gait. The evaluation of bone mineralization can be an important component of a diagnostic investigation of lameness. Lameness in growing pigs can cause an increase in morbidity and mortality, which leads to economic losses and animal welfare concerns for producers. Calcium and P are the primary minerals in skeletal tissue and their deficiency is considered to be one of the causes of lameness. To evaluate bone mineralization, it is important to know the differences between methodologies used to determine bone ash and the expected differences between the bones analyzed. Furthermore, there has been limited data comparing bone mineralization and serum Ca and P concentrations between healthy pigs and those exhibiting clinical signs of illness (unhealthy). By removing the lipid in the bone (defatting) before the bone is ashed, variation across bones is decreased compared with not removing lipid before ashing (non-defatted). The reduction in variation across bones allows for more differences to be detected among dietary treatments and health statuses of pigs. The 10th rib is more sensitive to detect dietary differences using bone density than metacarpals, fibulas, and 2nd ribs. When comparing healthy vs. unhealthy pigs exhibiting clinical signs of illness, healthy pigs have increased defatted percentage bone ash and serum Ca, P, and vitamin D.

Phase 1 study of CAR-37 T cells in patients with relapsed or refractory CD37+ lymphoid malignancies.

We report on the first-in-human clinical trial using chimeric antigen receptor (CAR) T-cells targeting CD37, an antigen highly expressed in B- and T-cell malignancies (clinicaltrials.gov NCT04136275). Five patients with relapsed or refractory CD37+ lymphoid malignancies were enrolled and infused with autologous CAR-37 T-cells. CAR-37 T-cells expanded in the peripheral blood of all patients and, at peak, comprised >94% of the total lymphocytes in 4/5 patients. Tumor responses were observed in 4/5 patients, with 3 complete responses, 1 mixed response, and 1 patient whose disease progressed rapidly and with relative loss of CD37 expression. Three patients experienced prolonged and severe pancytopenia, and in two of these patients, efforts to ablate CAR-37 T-cells (which were engineered to co-express truncated EGFR) with cetuximab, were unsuccessful. Hematopoiesis was restored in these two patients following allogeneic hematopoietic stem cell transplantation. No other severe, non-hematopoietic toxicities occurred. We investigated the mechanisms of profound pancytopenia and did not observe activation of CAR-37 T-cells in response to hematopoietic stem cells in vitro or hematotoxicity in humanized models. Patients with pancytopenia had sustained high levels of IL-18, with low levels of IL-18 binding protein in their peripheral blood. IL-18 levels were significantly higher in CAR-37-treated patients relative to both cytopenic and non-cytopenic cohorts of CAR-19-treated cohorts of patients. In conclusion, CAR-37 T-cells exhibited anti-tumor activity, with significant CAR expansion and cytokine production. CAR-37 T-cells may be an effective therapy in hematologic malignancies as a bridge to hematopoietic stem cell transplant.

Diagnostic survey of analytical methods used to determine bone mineralization in pigs.

Pigs from 64 commercial sites across 14 production systems in the Midwest United States were evaluated for baseline biological measurements used to determine bone mineralization. There were three pigs selected from each commercial site representing: 1) a clinically normal pig (healthy), 2) a pig with evidence of clinical lameness (lame), and 3) a pig from a hospital pen that was assumed to have recent low feed intake (unhealthy). Pigs ranged in age from nursery to market weight, with the three pigs sampled from each site representing the same age or phase of production. Blood, urine, metacarpal, fibula, 2nd rib, and 10th rib were collected and analyzed. Each bone was measured for density and ash (defatted and non-defatted technique). A bone × pig type interaction (P < 0.001) was observed for defatted and non-defatted bone ash and density. For defatted bone ash, there were no differences among pig types for the fibulas, 2nd rib, and 10th rib (P > 0.10), but metacarpals from healthy pigs had greater (P < 0.05) percentage bone ash compared to unhealthy pigs, with the lame pigs intermediate. For non-defatted bone ash, there were no differences among pig types for metacarpals and fibulas (P > 0.10), but unhealthy pigs had greater (P < 0.05) non-defatted percentage bone ash for 2nd and 10th ribs compared to healthy pigs, with lame pigs intermediate. Healthy and lame pigs had greater (P < 0.05) bone density than unhealthy pigs for metacarpals and fibulas, with no difference observed for ribs (P > 0.10). Healthy pigs had greater (P < 0.05) serum Ca and 25(OH)D3 compared to unhealthy pigs, with lame pigs intermediate. Healthy pigs had greater (P < 0.05) serum P compared to unhealthy and lame pigs, with no differences between the unhealthy and lame pigs. Unhealthy pigs excreted significantly more (P < 0.05) P and creatinine in the urine compared to healthy pigs with lame pigs intermediate. In summary, there are differences in serum Ca, P, and vitamin D among healthy, lame, and unhealthy pigs. Differences in bone mineralization among pig types varied depending on the analytical procedure and bone, with a considerable range in values within pig type across the 14 production systems sampled.

There is little literature or data comparing bone diagnostic results for healthy, lame, and unhealthy pigs. Typically, diagnosticians assessing clinical lameness cases in pigs will measure bone mineralization along with histopathological evaluation to diagnose and assess the severity of metabolic bone disease. Bone ash is the primary method to determine bone mineralization, with the removal of the lipid in the bone (defatting) before the bone is ashed, compared to not removing the lipid before the ashing (non-defatted). Defatting the bone reduces the amount of variation across the bones compared to non-defatting. In this diagnostic survey, there was no difference among the healthy, lame, or unhealthy pigs when comparing defatted bone ash, however, unhealthy pigs had an increased bone ash percentage compared to the healthy and lame pigs when the bones were assessed using the non-defatted procedure. There was variation across production systems and pig types for serum vitamin D. When comparing the pig types, healthy pigs had increased serum Ca, P, and vitamin D [25(OH)D3] compared to the unhealthy pigs, with the lame pigs intermediate.

Optimization of a flow cytometry test for routine monitoring of B cell maturation antigen targeted CAR in peripheral blood.

Chimeric antigen receptor (CAR) modified T cell therapies targeting BCMA have displayed impressive activity in the treatment of multiple myeloma. There are currently two FDA licensed products, ciltacabtagene autoleucel and idecabtagene vicleucel, for treating relapsed and refractory disease. Although correlative analyses performed by product manufacturers have been reported in clinical trials, there are limited options for reliable BCMA CAR T detection assays for physicians and researchers looking to explore it as a biomarker for clinical outcome. Given the known association of CAR T cell expansion kinetics with toxicity and response, being able to quantify BCMA CAR T cells routinely and accurately in the blood of patients can serve as a valuable asset. Here, we optimized an accurate and sensitive flow cytometry test using a PE-conjugated soluble BCMA protein, with a lower limit of quantitation of 0.19% of CD3+ T cells, suitable for use as a routine assay for monitoring the frequency of BCMA CAR T cells in the blood of patients receiving either ciltacabtagene autoleucel or idecabtagene vicleucel.

TaqMan Real-Time PCR Assay for Specific Detection and Differentiation of Xanthomonas translucens pv. undulosa from Other Pathovars Targeting a Recombination Mediator Gene, recF.

Bacterial leaf streak and black chaff diseases of wheat caused by Xanthomonas translucens pv. undulosa is becoming a major constraint to growers and trade since it is seedborne. Molecular tools for specific detection/differentiation of pv. undulosa are lacking. We report the development of a TaqMan real-time PCR for specific detection/identification of pv. undulosa targeting the recombination mediator gene (recF). Analysis of the complete recF (1,117 bp) sequences identified the gene as a reliable phylogenetic marker for identification of pv. undulosa, differentiating it from the other pathovars; recF-based sequence homology values among the 11 pathovars correlated well with genome-based DNA-DNA hybridization values. The discriminatory power of recF to differentiate pv. undulosa from the other pathovars is due to nucleotide polymorphic positions. We used these nucleotide polymorphisms to develop a TaqMan PCR for specific detection of pv. undulosa. The specificity of the assay was validated using 67 bacterial and fungal/oomycete strains. The selected primers and the double-quenched FAM-labeled TaqMan probe were specific for the detection of 11 pv. undulosa/secalis strains. The 56 strains of other X. translucens pathovars (n = 39) and non-Xanthomonas spp. (n = 17) did not exhibit any detectable fluorescence. Also, greenhouse-inoculated and naturally infected wheat leaf samples showed positive reactions for the presence of pv. undulosa DNA but not healthy control plants. The TaqMan assay reliably detected as low as 1-pg DNA amount and 10 colony forming units of the target pathogen per reaction. This TaqMan assay could be useful to regulatory agencies with economic benefits to wheat growers.

Uncovering the history of recombination and population structure in western Canadian stripe rust populations through mating type alleles.

BACKGROUND: The population structure of crop pathogens such as Puccinia striiformis f. sp. tritici (Pst), the cause of wheat stripe rust, is of interest to researchers looking to understand these pathogens on a molecular level as well as those with an applied focus such as disease epidemiology. Cereal rusts can reproduce sexually or asexually, and the emergence of novel lineages has the potential to cause serious epidemics such as the one caused by the 'Warrior' lineage in Europe. In a global context, Pst lineages in Canada were not well-characterized and the origin of foreign incursions was not known. Additionally, while some Pst mating type genes have been identified in published genomes, there has been no rigorous assessment of mating type diversity and distribution across the species. RESULTS: We used a whole-genome/transcriptome sequencing approach for the Canadian Pst population to identify lineages in their global context and evidence tracing foreign incursions. More importantly: for the first time ever, we identified nine alleles of the homeodomain mating type locus in the worldwide Pst population and show that previously identified lineages exhibit a single pair of these alleles. Consistently with the literature, we find only two pheromone receptor mating type alleles. We show that the recent population shift from the 'PstS1' lineage to the 'PstS1-related' lineage is also associated with the introduction of a novel mating type allele (Pst-b3-HD) to the Canadian population. We also show evidence for high levels of mating type diversity in samples associated with the Himalayan center of diversity for Pst, including a single Canadian race previously identified as 'PstPr' (probable recombinant) which we identify as a foreign incursion, most closely related to isolates sampled from China circa 2015. CONCLUSIONS: These data describe a recent shift in the population of Canadian Pst field isolates and characterize homeodomain-locus mating type alleles in the global Pst population which can now be utilized in testing several research questions and hypotheses around sexuality and hybridization in rust fungi.

Multi-locus genome-wide association studies reveal the genetic architecture of Fusarium head blight resistance in durum wheat.

Durum wheat is more susceptible to Fusarium head blight (FHB) than other types or classes of wheat. The disease is one of the most devastating in wheat; it reduces yield and end-use quality and contaminates the grain with fungal mycotoxins such as deoxynivalenol (DON). A panel of 265 Canadian and European durum wheat cultivars, as well as breeding and experimental lines, were tested in artificially inoculated field environments (2019-2022, inclusive) and two greenhouse trials (2019 and 2020). The trials were assessed for FHB severity and incidence, visual rating index, Fusarium-damaged kernels, DON accumulation, anthesis or heading date, maturity date, and plant height. In addition, yellow pigment and protein content were analyzed for the 2020 field season. To capture loci underlying FHB resistance and related traits, GWAS was performed using single-locus and several multi-locus models, employing 13,504 SNPs. Thirty-one QTL significantly associated with one or more FHB-related traits were identified, of which nine were consistent across environments and associated with multiple FHB-related traits. Although many of the QTL were identified in regions previously reported to affect FHB, the QTL QFhb-3B.2, associated with FHB severity, incidence, and DON accumulation, appears to be novel. We developed KASP markers for six FHB-associated QTL that were consistently detected across multiple environments and validated them on the Global Durum Panel (GDP). Analysis of allelic diversity and the frequencies of these revealed that the lines in the GDP harbor between zero and six resistance alleles. This study provides a comprehensive assessment of the genetic basis of FHB resistance and DON accumulation in durum wheat. Accessions with multiple favorable alleles were identified and will be useful genetic resources to improve FHB resistance in durum breeding programs through marker-assisted recurrent selection and gene stacking.

The effect of bone and analytical methods on the assessment of bone mineralization response to dietary phosphorus, phytase, and vitamin D in nursery pigs.

A total of 360 pigs (DNA 600 × 241, DNA; initially 11.9 ±â€…0.56 kg) were used in a 28-d trial to evaluate the effects of different bones and analytical methods on the assessment of bone mineralization response to dietary P, vitamin D, and phytase in nursery pigs. Pens of pigs (six pigs per pen) were randomized to six dietary treatments in a randomized complete block design with 10 pens per treatment. Dietary treatments were designed to create differences in bone mineralization and included: (1) 0.19% standardized total tract digestibility (STTD) P (deficient), (2) 0.33% STTD P (NRC [2012] requirement) using monocalcium phosphate, (3) 0.33% STTD P including 0.14% release from phytase (Ronozyme HiPhos 2700, DSM Nutritional Products, Parsippany, NJ), (4) 0.44% STTD P using monocalcium phosphate, phytase, and no vitamin D, (5) diet 4 with vitamin D (1,653 IU/kg), and (6) diet 5 with an additional 50 µg/kg of 25(OH)D3 (HyD, DSM Nutritional Products, Parsippany, NJ) estimated to provide an additional 2,000 IU/kg of vitamin D3. After 28 d on feed, eight pigs per treatment were euthanized for bone (metacarpal, 2nd rib, 10th rib, and fibula), blood, and urine analysis. The response to treatment for bone density and ash was dependent upon the bone analyzed (treatment × bone interaction for bone density, P = 0.044; non-defatted bone ash, P = 0.060; defatted bone ash, P = 0.068). Thus, the response related to dietary treatment differed depending on which bone (metacarpal, fibula, 2nd rib, or 10th rib) was measured. Pigs fed 0.19% STTD P had decreased (P < 0.05) bone density and ash (non-defatted and defatted) for all bones compared to 0.44% STTD P, with 0.33% STTD P generally intermediate or similar to 0.44% STTD P. Pigs fed 0.44% STTD P with no vitamin D had greater (P < 0.05) non-defatted fibula ash compared to all treatments other than 0.44% STTD P with added 25(OH)D3. Pigs fed diets with 0.44% STTD P had greater (P < 0.05) defatted second rib ash compared to pigs fed 0.19% STTD P or 0.33% STTD P with no phytase. In summary, bone density and ash responses varied depending on bone analyzed. Differences in bone density and ash in response to P and vitamin D were most apparent with fibulas and second ribs. There were apparent differences in the bone ash percentage between defatted and non-defatted bone. However, differences between the treatments remain consistent regardless of the analytic procedure. For histopathology, 10th ribs were more sensitive than 2nd ribs or fibulas for the detection of lesions.

Lameness is defined as impaired movement or deviation from normal gait. There are many factors that can contribute to lameness, including but not limited to: infectious disease, genetic and conformational anomaly, and toxicity that affects the bone, muscle, and nervous systems. Metabolic bone disease is another cause of lameness in swine production and can be caused by inappropriate levels of essential vitamins or minerals. To understand and evaluate bone mineralization, it is important to understand the differences in diagnostic results between different bones and analytical techniques. Historically, percentage bone ash has been used as one of the procedures to assess metabolic bone disease as it measures the level of bone mineralization; however, procedures and results vary depending on the methodology and type of bone measured. Differences in bone density and ash in response to dietary P and vitamin D were most apparent in the fibulas and second ribs. There were apparent differences in the percentage of bone ash between defatted and non-defatted bone; however, the differences between the treatments remain consistent regardless of the analytic procedure. For histopathology, 10th ribs were more sensitive than 2nd ribs or fibulas for detection of lesions associated with metabolic bone disease.

Determining the phosphorus release curve for Smizyme TS G5 2,500 phytase from 500 to 2,500 FTU/kg in nursery pig diets.

A total of 320 pigs (Line 241 × 600, DNA, Columbus, NE; initially 11.9 ±â€…0.22 kg) were used in a 21-d growth study to determine the available P (aP) release curve for Smizyme TS G5 2,500 (Barentz, Woodbury, MN). At approximately 19 d of age, pigs were weaned, randomly allotted to pens, and fed common starter diets. Pigs were blocked by average pen body weight (BW) and randomly allotted to one of eight dietary treatments on day 18 postweaning, considered day 0 of the study. Dietary treatments were derived from a single basal diet and ingredients including phytase, monocalcium P, limestone, and sand were added to create the treatment diets. Treatments included three diets containing increasing inorganic P from monocalcium P (0.11%, 0.20%, and 0.28% aP), or five diets with increasing phytase (500, 1,000, 1,500, 2,000, or 2,500 FTU/kg) added to the diet containing 0.11% aP. All diets were corn-soybean meal-canola meal-based and were formulated to contain 1.24% standardized ileal digestibility Lys, 0.30% phytate P, and an analyzed Ca:P ratio of 1.10:1. Prior to the beginning of the study, all pigs were fed a diet containing 0.11% aP for a 2-d period (days 16 to 18 postweaning). At the conclusion of the study, one pig, closest to the mean weight of each pen, was euthanized and the right fibula, rib, and metacarpal were collected to determine bone ash, density, and total bone P. Bones were weighed while suspended in a vessel of water and the weights used to calculate bone density (Archimedes' principle). For bone ash, bones were processed using the non-defatted method. For the overall experimental period, pigs fed increasing inorganic P had increased (quadratic, P ≤ 0.033) average daily gain (ADG), average daily feed intake (ADFI), and final BW and a tendency for increased (quadratic, P ≤ 0.090) gain:feed ratio (G:F). Pigs fed increasing phytase had increased (quadratic, P ≤ 0.004) ADG, G:F, and final BW and increased (linear, P = 0.019) ADFI. For fibula, rib, and metacarpal characteristics, pigs fed increasing aP from inorganic P had increased (linear, P < 0.001) bone ash weight, percentage bone ash, bone density, and bone P concentration. Additionally, pigs fed increasing phytase had increased (linear or quadratic, P < 0.05) bone ash weight, percentage bone ash, bone density, and bone P. TheaP release curve generated for Smizyme TS G5 2,500 for percentage bone ash using data generated from all three bones is aP = (0.228 × FTU/kg) ÷ (998.065 + FTU/kg).

Impact of dietary analyzed calcium to phosphorus ratios and standardized total tract digestible phosphorus to net energy ratios on growth performance, bone, and carcass characteristics of pigs.

A total of 2,184 pigs (337 × 1,050, PIC; initially 12.4 ± 0.17 kg) were used in a 143-d study to evaluate the effects of feeding varying analyzed calcium to phosphorus ratios (Ca:P) at two standardized total tract digestible (STTD) phosphorus to net energy ratios (STTD P:NE). Pens of pigs (26 pigs per pen) were assigned to 1 of the 6 dietary treatments in a 2 × 3 factorial with main effects of STTD P:NE and Ca:P ratio. Diets consisted of two levels of STTD P:NE; High (1.80, 1.62, 1.43, 1.25, 1.10, and 0.99 g STTD P/Mcal NE from 11 to 22, 22 to 40, 40 to 58, 58 to 81, 81 to 104, and 104 to 129 kg, respectively); or Low (75% of the High levels), and three analyzed Ca:P ratios (0.90:1, 1.30:1, and 1.75:1). There were 14 pens per treatment. Diets were corn-soybean meal-based and contained a constant phytase concentration within each dietary phase with levels decreasing throughout the trial (phases 1 through 3, 500 FTU/kg, assumed release of 0.13% STTD P; phase 4, 400 FTU/kg, assumed release of 0.11% STTD P; phase 5, 290 FTU/kg, assumed release of 0.09% STTD P; and phase 6, 210 FTU/kg, assumed release of 0.07% STTD P). Overall, there was a Ca:P × STTD P:NE interaction (P < 0.05) observed for average daily gain (ADG), feed efficiency (G:F), final body weight (BW), hot carcass weight (HCW), bone mineral density, bone mineral content, and bone-breaking strength. When feeding Low STTD P:NE levels, increasing the analyzed Ca:P ratio decreased (linear, P < 0.001) ADG final BW, HCW, and tended to worsen G:F, bone mineral density, and bone mineral content (linear, P < 0.10). However, when feeding High STTD P:NE levels, increasing the analyzed Ca:P ratio significantly improved bone mineral content and bone mineral density (linear, P < 0.05), and tended to improve ADG and final BW (linear, P < 0.10) and G:F (quadratic P < 0.10). Additionally, increasing the analyzed Ca:P ratio worsened ADG, G:F, and bone mineralization with Low STTD P:NE but had marginal impacts when adequate STTD P:NE was fed.

Calcium (Ca) and phosphorus (P) are the most abundant minerals in the pig and are involved in lean tissue deposition and synthesis and maintenance of the skeletal structure. Swine diets are typically formulated with low margins of safety for P and excess P in the diet can lead to increased P excretion, which can result in negative environmental effects. To have an adequate utilization of both Ca and P, it is important to consider the Ca:P ratio when formulating pig diets. Research has shown that a wide Ca:P is detrimental to pig growth performance and bone mineralization when diets are low in STTD P. Therefore, the objective of this study was to evaluate the impact of varying Ca:P ratios fed at two levels of STTD P:NE on growth performance, bone, and carcass characteristics of pigs from 12 to 129 kg. When P levels were below requirement estimates, widening the Ca:P ratio from 0.90:1 to 1.75:1 reduced growth performance and bone mineralization; however, widening the Ca:P ratio improved performance and bone mineralization when P levels of the diet were above requirement estimates.

First Report of Bacterial Leaf Streak of Barley (Hordeum vulgare) Caused by Xanthomonas translucens pv. translucens in British Columbia, Canada.

Bacterial leaf streak (BLS) of barley is caused by the Gram-negative bacterial pathogen Xanthomonas translucens (Sapkota et al. 2020). In 2021, we observed multiple hill plots with BLS symptomatic plants in a barley stripe rust nursery in Vancouver, BC, Canada. We collected 29 leaf samples showing typical BLS symptoms (e.g. necrotic lesions; Fig. S1) and stored at 4 oC until bacterial isolation. Samples were surface-sterilized in 10% NaOCl for 20 sec and rinsed twice. About 1 cm2 of leaf tissue containing BLS characteristic lesions was macerated in 200 µL sterile H2O on a petri dish, incubated for 15 min, and 10 µl of the homogenates was streaked onto Wilbrink's - Boric Acid - Cephalexin (WBC) agar medium. Plates were incubated at 28-30 oC for 48 hrs. Four single colonies were obtained: BC10-1-2a (USask BC10-2a), BC10-1-2b (USask BC10-2b), UBC026 and UBC028. Colonies were grown in WBC broth and gDNA was extracted using E.Z.N.A. Bacterial DNA Kit (Omega Bio-Tek) or DNeasy Plant Pro Kit® (Qiagen) following manufacturer protocols. Genus-level identification was achieved using 16S rRNA sequencing with 27F/1492R primers (Lane 1991) of UBC026 (1,399 bp; NCBI # OP327375) and UBC028 (1,415 bp; NCBI #OP327376). Complete 16S rRNA sequences (1,533bp) of BC10-2a and BC10-2b (1,533 bp) were extracted from the draft whole-genome sequences (WGS) generated in this study. The 16S rRNA sequence homology values of 99.0-100% were recorded between the 4 strains. BLAST analyses of the 16S rRNA sequences to GenBank entries exhibited 99.5-100% similarity values (100% coverage) with the pathotype strains of Xtt DSM 18974T (LT604072) and X. translucens pv. undulosa (Xtu) CFBP 2055 (CP074361). Whole genomes of BC10-2a (JANUQY01) and BC10-2b (JANUQZ01) were sequenced (150-bp; reads 33.1 million; mean coverage 2125x) using NovaSeq Illumina, assembled (Unicycler v0.4.8; Wick et al. 2017) and analyzed to identify the strains to the species-level (Tambong et al. 2021). WGS of strains USask BC10-2a and USask BC10-2b exhibited genome-based DNA-DNA hybridization (dDDH; Meier-Kolthoff et al. 2013) and BLAST-based average nucleotide identity (ANIb; Richter et al. 2015) of 100%. The two strains also showed dDDH and ANIb of 90.4% (species-leel cut-off of 70%) and 98.780% and 98.80% (cut-off of 96%), respectively, with Xtt DSM 18974T (LT604072). In contrast, the WGS of BC10-2a and BC10-2b exhibited only 78.2% dDDH homology values with Xtu CFBP 2055T, suggesting that the strains are genetically more similar to Xtt. The assignment of these strains to Xtt is corroborated by phylogenomic analysis (Fig. S2; Meier-Kolthoff and Göker 2019) that showed the two strains clustering together (100% bootstrap) with the type strain DSM 18974T. These data suggest that these strains are taxonomically members of Xtt. Identification was also confirmed to the genus-level by LAMP assay using published X. translucens primers (Langlois et al. 2017). Pathovar-level identification was confirmed using a cbsA and S8.pep multiplex PCR diagnostic assay (Roman-Reyna et al. 2022). Koch's postulates were verified by greenhouse inoculation via leaf infiltration of UBC026 and UBC028 on 21-day old barley plants (line HB522) using an inoculum of 108 CFU ml-1 followed by re-isolation of the bacteria on WBC. The inoculated plants showed typical BLS symptoms similar to those observed in the field (Fig. S1). Water-inoculated plants had no symptoms. To our knowledge, this is the first published report of BLS of barley in British Columbia.

Systematic Characterization of Multi-Rust Resistance Genes from a 'Parula × Thatcher' Population with a High-Density Genetic Map.

Pyramiding multiple resistant genes has been proposed as the most effective way to control wheat rust diseases globally. Identifying the most effective pyramids is challenged by the large pool of rust resistance genes and limited information about their mechanisms of resistance and interactions. Here, using a high-density genetic map, a double haploid population, and multi-rust field testing, we aimed to systematically characterize the most effective gene pyramids for rust resistance from the durable multi-rust resistant CIMMYT cultivar Parula. We revealed that the Parula resistance gene pyramid contains Lr34/Yr18/Sr57 (Lr34), Lr46/Yr29/Sr58 (Lr46), Lr27/Yr30/Sr2 (Sr2), and Lr68. The efficacy, magnitude of effect, and interactions varied for the three rust diseases. A subpopulation mapping approach was applied to characterize the complex interactions of the resistance genes by controlling for the effect of Lr34. Using this approach, we found that Lr34 and Lr68 have a strong additive effect for leaf rust, whereas no additive effects were observed for any rusts between Lr34 and Lr46. Lr34 combined synergistically with Sr12 from Thatcher for stem rust, whereas the additive effect of Lr34 and Sr2 was dependent on the type of rust and environment. Two novel leaf rust quantitative trait loci (QTLs) from Parula were identified in this study, a stable QTL QLr-7BS and QLr-5AS, which showed Lr34 dependent expression. With these findings, we propose combining two to three high-value genes from Canadian wheat (e.g., Sr12 from Thatcher) with a foundational multi-adult plant resistance cassette for desirable and durable resistance to all three rusts in Canadian wheat.

Boys Round Here: The Relationship Between Masculine Honor Ideology, Aggressive Behavior, Race, and Regional Affiliation.

Masculine honor ideology (MHI) refers to a set of beliefs that dictate men must respond aggressively to threat or insult to maintain their ideal masculine reputation. The current study demonstrates the robust relationship between MHI and lifetime aggression outcomes in a national sample of men from the United States. It also details the regional prevalence of MHI and compares these rates across races and regions of the country. Participants included 896 adult United States men (Mage = 35.86, SD = 1.22) recruited on Amazon's Mechanical Turk. It was expected that the odds of endorsing past aggressive behavior and lifetime maladjustment would be increased by stronger adherence to MHI. This hypothesis was supported, and individuals who reported greater MHI adherence also had higher rates of lifetime aggression and maladjustment. Contrary to expectations, White, non-Hispanic men endorsed lower rates of MHI than did other men. Black men adhered more strongly to MHI than White and Hispanic men. It was also expected that men in the Southern and Western United States would endorse greater MHI in comparison to men in the Northeast United States. The hypothesis was only partially supported for White, non-Hispanic men, and it was associated with participant birthplace and their father's birthplace. There were no regional differences in MHI adherence related to the participants' mother's birthplace or where participants lived at survey completion. These findings suggest that MHI may spread more uniformly than prior research suggests and that MHI may have more nuanced cultural considerations that deserve continued empirical investigation.

Mental health of women living with HIV and its impact on child development in Andhra Pradesh, India.

Global literature examining the association between mental health of women living with HIV (WLWH) and child development is scarce. In this study, we examined the relationship between mothers' mental health and their children's social development outcomes 6 months later. Data for these analyses come from several waves of interviews of 600 WLWH in the South Indian state of Andhra Pradesh, India. These women were enrolled in a 2×2 factorial clinical trial designed to assess the impact of food supplementation and nutrition education, both in addition to ASHA support, on adherence to ART and improved health outcomes for the women and one of their children. They were assessed on food security, stigma, social support, quality of life, depressive symptoms and child development outcomes. Results of longitudinal GEE regression analysis indicate that mother's depressive symptoms were significantly negatively associated with child's social quotient 6 months later. These findings have important implications for targeted health interventions, integrating mental health, both for WLWH and their children in India.

Influence of Enogen Feed corn and conventional yellow dent corn in pelleted or meal-based diets on finishing pig performance and carcass characteristics.

Genetic modification of corn has enhanced the use of different corn hybrids in animal agriculture. Enogen Feed corn, developed by Syngenta Seeds (Downers Grove, IL), has potential for use in livestock diets due to increase α-amylase enzyme in the corn thus improving starch digestibility. In addition, the pelleting process also increases starch gelatinization which increases its digestibility by the pig, increasing growth rate and improving feed efficiency. Therefore, pelleting Enogen Feed corn might prove to provide a greater response in growth performance than conventional yellow dent corn. Thus, the objective of this experiment was to determine the effects of corn source and diet form on growth performance and carcass characteristics of finishing pigs. A total of 288 pigs (53.0 ± 0.5 kg) were used with eight pigs per pen and nine pens per treatment in a 72-d study. Treatments were arranged in a 2 × 2 factorial with main effects of corn source (Enogen Feed corn or conventional yellow dent corn) and diet form (meal or pellet). For overall (d 0 to 72) performance, no interactions between corn source and diet form were observed. There was a tendency (P < 0.10) for slightly improved average daily gain (ADG) and gain:feed ratio (G:F) for pigs fed conventional yellow dent corn compared to those fed Enogen Feed corn. For feed form, pigs fed pelleted diets had increased (P < 0.001) ADG and G:F compared to pigs fed meal diets. For carcass characteristics, pigs fed pelleted diets had increased hot carcass weight compared to pigs fed meal diets (P < 0.001). In summary, feeding pelleted diets to finishing pigs increased ADG and improved feed efficiency compared to those fed meal-based diets. There were no major differences between observed corn sources or interactions between corn source and diet form on growth performance.

Incursions of divergent genotypes, evolution of virulence and host jumps shape a continental clonal population of the stripe rust pathogen Puccinia striiformis.

Long-distance migration and host adaptation by transboundary plant pathogens often brings detrimental effects to important agroecosystems. Efficient surveillance as a basis for responding to the dynamics of such pathogens is often hampered by a lack of information on incursion origin, evolutionary pathways and the genetic basis of rapidly evolving virulence across larger timescales. Here, we studied these genetic features by using historical isolates of the obligate biotrophic pathogen Puccinia striiformis f. sp. tritici (Pst), which causes one of the most widespread and devastating diseases, stripe (yellow) rust, of wheat. Through a combination of genotypic, phenotypic and genomic analyses, we assigned eight Pst isolates representing putative exotic Pst incursions into Australia to four previously defined genetic groups, PstS0, PstS1, PstS10 and PstS13. We showed that isolates of an additional incursion of P. striiformis, known locally as P. striiformis f. sp. pseudo-hordei, had a new and unique multilocus SSR genotype (MLG). We provide results of overall genomic variation of representative Pst isolates from each genetic group by comparative genomic analyses. We showed that isolates within the PstS1 and PstS13 genetic groups are most distinct at the whole-genome variant level from isolates belonging to genetic group PstS0, whereas the isolate from the PstS10 genetic group is intermediate. We further explored variable gene content, including putative effectors, representing both shared but also unique genetic changes that have occurred following introduction, some of which may additionally account for local adaptation of these isolates to triticale. Our genotypic and genomic data revealed new genetic insights into the evolution of diverse phenotypes of rust pathogens following incursion into a geographically isolated continental region.

The influence of particle size of Enogen Feed corn and conventional yellow dent corn on nursery and finishing pig performance, carcass characteristics and stomach morphology.

Enogen Feed corn is a variety developed by Syngenta Seeds (Downers Grove, IL) that has been genetically modified to contain an α-amylase enzyme trait (SYT-EFC). Originally, Enogen feed corn was developed for the ethanol industry due to its reduction in viscosity of the corn mash, thus eliminating the need to add a liquid form of the α-amylase enzyme. However, there is a potential application for Enogen Feed corn to be used in livestock diets due to the increase in α-amylase enzyme potential to increase starch digestibility. A more common method of increasing starch digestibility in corn is to finely grind it to reduce particle size. This increases the surface area and allows for greater interaction with digestive enzymes. We hypothesized that pigs fed Enogen feed corn potentially could achieve similar gain:feed ratio (G:F) at larger particle sizes than conventional corn because of the differences in starch digestibility. In experiment 1, a total of 360 pigs (DNA 200 × 400, Columbus, NE; initially 6.6 ± 0.1 kg BW) were used with five pigs per pen and 12 pens per treatment. Treatments were arranged in a 2 × 3 factorial with main effects of corn source (Enogen Feed corn or conventional yellow dent corn) and ground corn particle size (300, 600, or 900 µm). Overall, there was a corn source × particle size interaction (linear, P = 0.027) for G:F. There was no effect due to particle size when pigs were fed conventional yellow dent corn, but in pigs fed Enogen Feed corn, G:F increased with decreasing particle size. Neither corn source nor particle size affected (P > 0.05) overall average daily gain (ADG) or average daily feed intake (ADFI). In experiment 2, a total of 323 pigs (241 × 600; DNA, Columbus, NE; initially 50.0 ± 1.3 kg) were used with nine pigs per pen and six pens per treatment. Treatments were identical as experiement 1. Overall, corn source had no effect on finishing pig ADG, ADFI or G:F. For corn particle size, ADG and G:F increased (linear, P < 0.014) and ADFI decreased (P = 0.043) as particle size decreased. For stomach morphology, there was a tendency for a corn source × particle size interaction (P = 0.055) for keratinization score with keratinization increasing linearly (P = 0.001) as particle size of the corn decreased for yellow dent corn with no change in keratinization score as particle size decreased for Enogen Feed corn. In summary, reducing corn particle size improved G:F with no major differences observed between corn sources for overall pig performance.

Influence of particle size of Enogen Feed corn and conventional yellow dent corn on lactating sow performance.

Enogen Feed corn is a variety developed by Syngenta Seeds (Downers Grove, IL) that has been genetically modified to contain an -amylase enzyme trait (SYT-EFC). Originally, Enogen feed corn was developed for the ethanol industry due to its properties for reducing the viscosity of its corn mash. There is potential application for Enogen Feed corn to be used in livestock diets due to the potential for the increase in - amylase enzyme to increase the starch digestibility. Because of this, it may be possible to increase the particle size of ground Enogen Feed corn and maintain the same starch digestibility as finely ground conventional yellow dent corn. Therefore, our hypothesis was that an interaction between corn source and particle size would exist such that the performance of sows fed fine ground conventional yellow dent corn would be similar to sows fed coarse ground Enogen Feed corn. A total of 107 sows (Line 241; DNA, Columbus, NE) across four batch farrowing groups were used to evaluate sow and litter performance. Treatments were arranged in a 2 2 factorial with main effects of corn source (Enogen Feed corn or conventional yellow dent corn) and ground corn particle size (600 or 900 m). From farrowing to weaning, there was a tendency for a corn source particle size interaction (P = 0.065) in sow body weight (BW) change. Sows fed 900 m Enogen Feed corn had decreased BW loss compared to sows fed other treatments, which were similar in weight loss. For sow average daily feed intake from farrowing to weaning, there was a corn source particle size interaction (P = 0.048) with sows fed 900 m conventional yellow dent corn having lower feed intake than the sows fed 600 m conventional yellow dent corn, whereas sows fed 900 m Enogen Feed corn had greater feed intake compared to the sows fed 600 m Enogen Feed corn. There was a tendency for a particle size main effect (P < 0.10) for litter average daily gain (ADG) and total litter gain, with sows fed corn ground to 600 m having increased litter ADG and total litter gain compared to sows fed corn ground to 900 m. In summary, there were few differences in sow or litter characteristics among those fed Enogen Feed corn or conventional yellow dent corn. Reducing particle size of both corn sources tended to increase litter ADG and weaning weights.