Stratification of Fusobacterium nucleatum by local health status in the oral cavity defines its subspecies disease association.

The ubiquitous inflammophilic oral pathobiont Fusobacterium nucleatum (Fn) is widely recognized for its strong association with inflammatory dysbiotic diseases and cancer. Fn is subdivided into four subspecies, which are historically considered functionally interchangeable in the oral cavity. To test this assumption, we analyzed patient-matched dental plaque and odontogenic abscess clinical specimens and examined whether an inflammatory environment selects for/against particular Fn subspecies. Dental plaque harbored a greater diversity of fusobacteria, with Fn. polymorphum dominating, whereas odontogenic abscesses were exceptionally biased for the largely uncharacterized organism Fn. animalis. Comparative genomic analyses revealed significant genotypic distinctions among Fn subspecies that correlate with their preferred ecological niches and support a taxonomic reassignment of each as a distinct Fusobacterium species. Despite originating as a low-abundance organism in dental plaque, Fn. animalis typically outcompetes other oral fusobacteria within the inflammatory abscess environment, which may explain its prevalence in other oral and extraoral diseases.

The prevalence of Fusobacterium nucleatum subspecies in the oral cavity stratifies by local health status.

The ubiquitous inflammophilic pathobiont Fusobacterium nucleatum is widely recognized for its strong association with a variety of human dysbiotic diseases such as periodontitis and oral/extraoral abscesses, as well as multiple types of cancer. F. nucleatum is currently subdivided into four subspecies: F. nucleatum subspecies nucleatum (Fn. nucleatum), animalis (Fn. animalis), polymorphum (Fn. polymorphum), and vincentii/fusiforme (Fn. vincentii). Although these subspecies have been historically considered as functionally interchangeable in the oral cavity, direct clinical evidence is largely lacking for this assertion. Consequently, we assembled a collection of oral clinical specimens to determine whether F. nucleatum subspecies prevalence in the oral cavity stratifies by local oral health status. Patient-matched clinical specimens of both disease-free dental plaque and odontogenic abscess were analyzed with newly developed culture-dependent and culture-independent approaches using 44 and 60 oral biofilm/tooth abscess paired specimens, respectively. Most oral cavities were found to simultaneously harbor multiple F. nucleatum subspecies, with a greater diversity present within dental plaque compared to abscesses. In dental plaque, Fn. polymorphum is clearly the dominant organism, but this changes dramatically within odontogenic abscesses where Fn. animalis is heavily favored over all other fusobacteria. Surprisingly, the most commonly studied F. nucleatum subspecies, Fn. nucleatum, is only a minor constituent in the oral cavity. To gain further insights into the genetic basis for these phenotypes, we subsequently performed pangenome, phylogenetic, and functional enrichment analyses of oral fusobacterial genomes using the Anvi'o platform, which revealed significant genotypic distinctions among F. nucleatum subspecies. Accordingly, our results strongly support a taxonomic reassignment of each F. nucleatum subspecies into distinct Fusobacterium species. Of these, Fn. animalis should be considered as the most clinically relevant at sites of active inflammation, despite being among the least characterized oral fusobacteria.

Who is in the driver's seat? Parvimonas micra: An understudied pathobiont at the crossroads of dysbiotic disease and cancer.

Recent advances in our understanding of microbiome composition at sites of inflammatory dysbiosis have triggered a substantial interest in a variety of historically understudied bacteria, especially among fastidious obligate anaerobes. A plethora of new evidence suggests that these microbes play outsized roles in establishing synergistic polymicrobial infections at many different sites in the human body. Parvimonas micra is a prime example of such an organism. Despite being almost completely uncharacterized at the genetic level, it is one of the few species commonly detected in abundance at multiple mucosal sites experiencing either chronic or acute inflammatory diseases, and more recently, it has been proposed as a discriminating biomarker for multiple types of malignancies. In the absence of disease, P. micra is commonly found in low abundance, typically residing within the oral cavity and gastrointestinal tract. P. micra exhibits the typical features of an inflammophilic organism, meaning its growth actually benefits from active inflammation and inflammatory tissue destruction. In this mini-review, we will describe our current understanding of this underappreciated but ubiquitous pathobiont, specifically focusing upon the role of P. micra in polymicrobial inflammatory dysbiosis and cancer as well as the key emerging questions regarding its pathobiology. Through this timely work, we highlight Parvimonas micra as a significant driver of disease and discuss its unique position at the crossroads of dysbiosis and cancer.

Frequency of maternal supplementation of energy and protein during late gestation modulates preweaning growth of their beef offspring.

This study evaluated the effects of decreasing the frequency of dried distillers grains (DDG) supplementation during third trimester of gestation on cow physiology and offspring preweaning growth. At 201 ± 7 d prepartum (day 0 of the study), 120 Brangus crossbred cows were stratified by body weight (BW = 543 ± 53 kg) and body condition score (BCS = 5.47 ± 0.73), and then assigned randomly to 1 of 20 bahiagrass (Paspalum notatum) pastures (six cows and 4.7 ha/pasture). Treatments were randomly assigned to pastures (five pastures/treatment) and consisted of cows offered no DDG supplementation (NOSUP) or precalving supplementation of DDG dry matter at 1 kg/cow daily (7×), 2.33 kg/cow every Monday, Wednesday, and Friday (3×), or 7 kg/cow every Monday (1×) from day 0 to 77. All cows assigned to DDG supplementation received the same total amount of DDG dry matter (77 kg/cow) from day 0 to 77. All cow-calf pairs were managed similarly from day 77 until calf weaning (day 342). Supplementation frequency did not impact (P ≥ 0.16) any forage or cow reproduction data. Cow BCS on days 77, 140, and 342 did not differ among 1×, 3×, and 7× cows (P ≥ 0.29) but all supplemented cows, regardless of supplementation frequency, had greater BCS on days 77, 140, and 342 compared to NOSUP cows (P ≤ 0.04). Cows offered 1× supplementation had greater plasma concentrations of IGF-1 on days 35 and 140 compared to NOSUP, 3× and 7× cows (P ≤ 0.04), whereas 3× and 7× cows had greater plasma concentrations of IGF-1 on day 35 compared to NOSUP cows (P ≤ 0.005). Average plasma concentrations of glucose did not differ among 1×, 3×, and 7× cows (P ≥ 0.44), but all supplemented cows had greater plasma concentrations of glucose compared to NOSUP cows (P ≤ 0.05). Birth BW of the first offspring did not differ between 3× and 7× calves (P = 0.54) but both groups were heavier at birth compared to NOSUP calves (P ≤ 0.05). On day 342, calves born from 7× cows were the heaviest (P ≤ 0.05), whereas calves born from 1× and 3× cows had similar BW (P = 0.97) but both groups were heavier compared to calves born from NOSUP cows (P ≤ 0.05). In summary, decreasing the frequency of DDG supplementation, from daily to one or three times weekly, during third trimester of gestation of beef cows did not impact cow BCS but altered maternal plasma concentrations of IGF-1 and glucose, leading to reduced offspring preweaning growth.

Addition of açai oil during the close-up dry period of Holstein cows improves colostrum quality and immune responses of their calves.

This study evaluated of the effects of açai oil during the close-up dry period of Holstein cows on colostrum quality, as well as on the immune and antioxidant responses of their calves. Sixteen multiparous cows were assigned randomly to two treatments: 1) CONTROL (n = 8) - 4.48% of soybean oil/concentrate; 2) AÇAI (n =8) - 4.48% of açai oil/concentrate. Cows fed with açai oil had greater (P≤0.04) colostrum concentrations of immunoglobulins (Ig) G (1st and 2nd milking), IgG heavy chains, IgA (only at 1st milking), alpha-lactalbumin (1st milking), total protein, and antioxidant capacity against peroxyl radicals (only at 1st milking). Cows fed with açai oil had greater serum concentrations of globulin (only on the day of calving) and total protein (only on the day of calving) (P = 0.03). Calves born of cows fed with açai oil had greater serum concentrations of total protein (only 24 and 48 h after calving) and serum concentration of IgG heavy chain (only 24 h after calving) and globulin (only 24 and 48 h after calving) (P = 0.01). These data suggest that the addition of açai oil in the cow feed during the close-up dry period boosted immunity in their calves by altering the composition of colostrum.

Stair-step strategy and immunomodulatory feed ingredient supplementation for grazing heat-stressed Bos indicus-influenced beef heifers.

On day 0 of years 1 and 2, 64 Brangus crossbred heifers per year were stratified by initial body weight (BW) and age (mean = 257 ± 20 kg and 271 ± 22 d) and allocated into 16 bahiagrass (Paspalum notatum) pastures (4 heifers per pasture per yr). Treatments were randomly allotted to pastures in a 2 × 2 factorial arrangement of treatments (4 pastures per treatment per yr). Treatments consisted of concentrate dry matter (DM) supplementation at 1.50% of BW from days 0 to 100 (CON) or concentrate DM supplementation at 1.05% of BW from days 0 to 49 and 1.95% of BW from days 50 to 100 (SST). Then, each respective supplementation strategy was added or not with immunomodulatory feed ingredients from days 0 to 100 (OMN; 4 g/45 kg of BW). Heifers were assigned to an estrus synchronization protocol from days 100 to 114. Heifers detected in estrus from days 111 to 114 were inseminated (AI) 12 h after estrus detection. Heifers not detected in estrus were timed AI on day 114. All heifers were exposed to Angus bulls from days 120 to 210 (1 bull per pasture). Effects of supplementation strategy × OMN inclusion × hour were detected (P < 0.0001) only for intravaginal temperature from days 26 to 30, which were the least (P ≤ 0.03) for SST heifers offered OMN supplementation and did not differ (P ≥ 0.17) among all remaining treatments from 0830 to 1600 hours. Effects of supplementation strategy × OMN inclusion and OMN inclusion were not detected (P ≥ 0.12) for any variable, except for the percentage of heifers detected in estrus, which was greater (P = 0.01) for heifers supplemented with vs. without OMN. Total concentrate DM offered from days 0 to 100 and heifer BW on days 0 and 56 did not differ (P ≥ 0.49) between CON and SST heifers, but SST heifers were heavier (P ≤ 0.01) on days 100 and 210 compared with CON heifers. Body surface temperature on day 25 and plasma IGF-1 concentrations on day 75 were greater (P ≤ 0.04) for SST vs. CON heifers. Percentage of pubertal heifers, heifers detected in estrus, and pregnancy to AI did not differ (P = 0.36) between SST and CON heifers but the final pregnancy percentage was greater (P = 0.04) for SST vs. CON heifers. Thus, OMN supplementation decreased the intravaginal temperature of SST heifers but failed to improve their growth and reproduction, whereas the SST strategy improved body thermoregulation, growth, and final pregnancy percentage of heat-stressed Bos indicus-influenced beef heifers compared with a constant concentrate supplementation strategy.

In Bos taurus beef heifers, altering the timing of body weight (BW) growth pattern either reduced feed costs without decreasing reproduction or enhanced reproduction without increasing feed costs. Moreover, supplementation of OmniGen-AF (OMN, a patented immunomodulatory feed ingredient) decreased internal body temperature in dairy and beef B. taurus cattle, with variable impacts on growth and reproduction. Combining both nutritional strategies for Bos indicus-beef heifers developed under heat stress conditions of tropical and subtropical environments has not been reported in the literature yet and was the main objective of the present study. For 100 d before the breeding season, heifers received either a constant supplementation amount or stair-step (SST) supplementation strategy (50 d of low followed by 50 d of high supplement amount), with or without OMN inclusion. Overall, OMN supplementation alleviated the internal body temperature of heifers but did not improve their growth and reproduction, whereas the SST strategy increased BW gain and final pregnancy percentage of B. indicus-influenced beef heifers under heat stress conditions.

Development of the First Tractable Genetic System for Parvimonas micra, a Ubiquitous Pathobiont in Human Dysbiotic Disease.

Parvimonas micra is a Gram-positive obligate anaerobe and a typical member of the human microbiome. P. micra is among the most highly enriched species at numerous sites of mucosal dysbiotic disease and is closely associated with the development of multiple types of malignant tumors. Despite its strong association with disease, surprisingly little is known about P. micra pathobiology, which is directly attributable to its longstanding genetic intractability. To address this problem, we directly isolated a collection of P. micra strains from odontogenic abscess clinical specimens and then screened these isolates for natural competence. Amazingly, all of the P. micra clinical isolates exhibited various levels of natural competence, including the reference strain ATCC 33270. By exploiting this ability, we were able to employ cloning-independent methodologies to engineer and complement a variety of targeted chromosomal genetic mutations directly within low-passage-number clinical isolates. To develop a tractable genetic system for P. micra, we first adapted renilla-based bioluminescence for highly sensitive reporter studies. This reporter system was then applied for the development of the novel Theo+ theophylline-inducible riboswitch for tunable gene expression studies over a broad dynamic range. Finally, we demonstrate the feasibility of generating mariner-based transposon sequencing (Tn-seq) libraries for forward genetic screening in P. micra. With the availability of a highly efficient transformation protocol and the current suite of genetic tools, P. micra should now be considered a fully genetically tractable organism suitable for molecular genetic research. The methods presented here provide a clear path to investigate the understudied role of P. micra in polymicrobial infections and tumorigenesis. IMPORTANCE Parvimonas micra is among the most highly enriched species at numerous sites of mucosal dysbiotic disease and is closely associated with numerous cancers. Despite this, little is known about P. micra pathobiology, which is directly attributable to its longstanding genetic intractability. In this study, we provide the first report of P. micra natural competence and describe the only tractable genetic system for this species. The methods presented here will allow for the detailed study of P. micra and its role in infection and tumorigenesis.

Effects of maternal winter vs. year-round supplementation of protein and energy on postnatal growth, immune function, and carcass characteristics of Bos indicus-influenced beef offspring.

This 2-yr study evaluated the effects of winter vs. year-round supplementation of Bos indicus-influenced beef cows on cow reproductive performance and impact on their offspring. On day 0 of each year (approximately day 122 ± 23 of gestation), 82 to 84 mature Brangus cows/yr were stratified by body weight (BW; 475 ± 67 kg) and body condition score (BCS; 4.85 ± 0.73) and randomly assigned to 1 of 6 bahiagrass (Paspalum notatum) pastures (13 to 14 cows/pasture). Treatments were randomly assigned to pastures consisting of winter supplementation with molasses + urea (WMOL), or year-round supplementation with molasses + urea (YMOL) or wheat middling-based range cubes (YCUB). Total yearly supplement DM amount was 272 kg/cow and supplements were formulated to be isocaloric and isonitrogenous (75% TDN and 20% CP). On day 421 (weaning; approximately 260 ± 24 d of age), 33 to 35 steers/yr were vaccinated against parainfluenza-3 (PI3) and bovine viral diarrhea virus type 1 (BVDV-1) and transported 1,193 km to a feedlot. Steers were penned according to maternal pasture and managed similarly until slaughter. Data were analyzed using the MIXED and GLIMMIX procedures of SAS. On day 217 (start of breeding season), BCS was greater (P = 0.01) for YMOL than WMOL cows, whereas BCS of YCUB did not differ (P ≥ 0.11) to both WMOL and YMOL cows. The percentage of cows that calved, calving date, birth BW, and preweaning BW of the first offspring did not differ (P ≥ 0.22) among maternal treatments. Plasma cortisol concentrations were greater (P ≤ 0.001) for YCUB steers at feedlot arrival (day 422) than WMOL and YMOL steers. Moreover, YCUB steers had greater (P = 0.02) and tended (P = 0.08) to have greater plasma concentrations of haptoglobin compared to WMOL and YMOL steers, respectively. Antibody titers against PI3 and BVDV-1 viruses did not differ (P ≥ 0.25) among maternal treatments. Steer BW at feedlot exit was greater (P ≤ 0.05) for YMOL and WMOL than YCUB steers. However, feedlot DMI did not differ (P ≥ 0.37) by maternal treatment. Hot carcass weight, yield grade, LMA, and marbling did not differ (P ≥ 0.14) among maternal treatments. Percentage of steers that graded low choice was enhanced (P ≤ 0.05) for WMOL and YCUB than YMOL steers. Maternal year-round supplementation of range cubes or molasses + urea either did not impact or decrease growth, immune function, and carcass characteristics of the offspring when compared with maternal supplementation of molasses + urea during winter only.

Timing of maternal supplementation of dried distillers grains during late gestation influences postnatal growth, immunocompetence, and carcass characteristics of Bos indicus-influenced beef calves.

This 2-yr study investigated the timing of dried distillers grains (DDG) supplementation during the third trimester of gestation of Bos indicus-influenced beef cows and its impact on their offspring performance. On day 0 of each year (84 d before calving), Brangus cows (n = 84/yr; cow age = 8 ± 3 yr) were stratified by initial body weight (BW; 482 ± 75 kg) and body condition score (BCS; 5.3 ± 0.8) and assigned randomly to one of six bahiagrass (Paspalum notatum) pastures (experimental units; 14 cows/pasture). Treatments were assigned randomly to pasture (2 pastures/treatment/yr) and consisted of no prepartum supplementation (CON), 2 kg/d of DDG from day 0 to 42 (LATE42), or 1 kg/d of DDG from day 0 to 84 (LATE84). Following calving (day 84), cow-calf pairs remained in their respective pastures, and cows were offered sugarcane molasses + urea (1.82 kg of dry matter/cow/d) from day 85 until the end of the breeding season (day 224). On day 347, steer calves (n = 38/yr; 11 to 15 steers/treatment/yr) were weaned and transported to the feedlot (1,193 km). Steers were penned according to cow prepartum pasture and managed similarly until the time of harvest. BCS at calving was greater (P < 0.01) for LATE42 and LATE84 vs. CON cows but did not differ (P = 0.16) between LATE42 and LATE84 cows. Calving date, calving percentage, and birth BW of the first offspring did not differ (P ≥ 0.22) among treatments. However, LATE42 cows calved their second offspring 8 d earlier (P = 0.04) compared with CON and LATE84 cows. At weaning (first offspring), LATE84 calves were the heaviest (P ≤ 0.05), CON calves were the lightest, and LATE42 calves had intermediate BW (P ≤ 0.05). Steer plasma concentrations of cortisol and haptoglobin and serum bovine viral diarrhea virus type-1 titers did not differ (P ≥ 0.21) between treatments. Steer serum parainfluenza-3 titers were greater (P = 0.03) for LATE42 vs. CON steers, tended to be greater (P = 0.10) for LATE84 compared with CON steers, and did not differ (P = 0.38) between LATE42 and LATE84 steers. Steer feedlot BW, average daily gain, dry matter intake, and hot carcass weight did not differ (P ≥ 0.36) between treatments. Marbling and the percentage of steers grading choice were greater (P ≤ 0.04) for LATE42 vs. CON steers, whereas LATE84 steers were intermediate. In summary, different timing of DDG supplementation during the third trimester of gestation could be explored to optimize cow BCS and offspring preweaning growth and carcass quality.

This 2-yr study evaluated the effect of the timing of dried distillers grains (DDG) supplementation during the last trimester of gestation in Bos indicus-influenced beef cows and the subsequent impact on their offspring. Brangus cows were allocated to one of the three prepartum treatments consisting of no prepartum supplementation, 2 kg/d of DDG for the first half of the last trimester of gestation, or 1 kg/d of DDG for the entire length of the last trimester of gestation. Prepartum supplementation, regardless of supplementation length, improved cow body condition scores at the time of calving. Calf birth weights were not affected by prepartum maternal treatment. Calves born to cows that received prepartum supplementation had greater weaning weight compared with no prepartum supplementation. However, weaning weights were improved to the greatest extent when calves were born to cows that received supplementation for the entire length of late gestation. Steer antibody response to parainfluenza-3 was improved with prepartum maternal supplementation, regardless of supplementation length. Furthermore, concentrating the total amount of supplement offered to the first half of the last trimester of gestation improved marbling and increased the percentage of steers grading choice compared with no prepartum supplementation or supplementation during the entire last trimester.

Effects of maternal gestational diet, with or without methionine, on muscle transcriptome of Bos indicus-influenced beef calves following a vaccine-induced immunological challenge.

Maternal nutrition during gestation can cause epigenetic effects that translate to alterations in gene expression in offspring. This 2-year study employed RNA-sequencing technology to evaluate the pre- and post-vaccination muscle transcriptome of early-weaned Bos indicus-influenced beef calves born from dams offered different supplementation strategies from 57 ± 5 d prepartum until 17 ± 5 d postpartum. Seventy-two Brangus heifers (36 heifers/yr) were stratified by body weight and body condition score and assigned to bahiagrass pastures (3 heifers/pasture/yr). Treatments were randomly assigned to pastures and consisted of (i) no pre- or postpartum supplementation (NOSUP), (ii) pre- and postpartum supplementation of protein and energy using 7.2 kg of dry matter/heifer/wk of molasses + urea (MOL), or (iii) MOL fortified with 105 g/heifer/wk of methionine hydroxy analog (MOLMET). Calves were weaned on d 147 of the study. On d 154, 24 calves/yr (8 calves/treatment) were randomly selected and individually limit-fed a high-concentrate diet until d 201. Calves were vaccinated on d 160. Muscle biopsies were collected from the same calves (4 calves/treatment/day/yr) on d 154 (pre-vaccination) and 201 (post-vaccination) for gene expression analysis using RNA sequencing. Molasses maternal supplementation led to a downregulation of genes associated with muscle cell differentiation and development along with intracellular signaling pathways (e.g., Wnt and TGF-ß signaling pathway) compared to no maternal supplementation. Maternal fortification with methionine altered functional gene-sets involved in amino acid transport and metabolism and the one-carbon cycle. In addition, muscle transcriptome was impacted by vaccination with a total of 2,396 differentially expressed genes (FDR ≤ 0.05) on d 201 vs. d 154. Genes involved in cell cycle progression, extracellular matrix, and collagen formation were upregulated after vaccination. This study demonstrated that maternal supplementation of energy and protein, with or without, methionine has long-term implications on the muscle transcriptome of offspring and potentially influence postnatal muscle development.

Differences in copper and selenium metabolism between Angus (Bos taurus) and Brahman (Bos indicus) cattle.

A 2-yr study was conducted at the Range Cattle Research and Education Center, University of Florida - Institute of Food and Agricultural Sciences (IFAS) (Ona, FL), to evaluate differences in the metabolism of Cu and Se of Angus (Bos taurus) and Brahman (Bos indicus) cattle. Thirty-two pregnant beef cows (n = 8 Brahman and 8 Angus/yr) were enrolled in the study in the first trimester of gestation. This study consisted of three phases: 1) restriction (day 0 to 90), 2) supplementation (day 91 to 150), and 3) calving. During all three phases, cows were individually fed and housed in partially covered drylot pens. During the restriction and supplementation phases, cows were provided a 1.5 kg/d of a grain-based concentrate supplement, which was fortified with flowers of S (50 g of supplemental S/cow daily; restriction phase) or Cu and Se (100 and 3 mg/d of Cu and Se, respectively; supplementation phase). Blood and liver samples were collected from all cows at 30 d intervals and from both cows and calves within 24 h of calving. Colostrum and milk samples were collected at calving and 7 d after birth. All data were analyzed using the MIXED procedure of SAS, where cow and calf were the experimental unit. During the restriction phase, a breed × day effect (P = 0.03) was observed where Brahman had greater liver Cu concentration than Angus cows in all sampling days. For liver Se concentration, a tendency (P = 0.07) for a breed effect was observed where Angus cows tended to have greater liver Se concentration than Brahman. During the supplementation phase, breed (P < 0.001) and day (P < 0.01) effects were observed, where Brahman cows had greater liver Cu concentration than Angus. For liver Se concentration, a day effect (P < 0.001) was observed, where liver Se concentration increased (P < 0.001) from day 90 to 120 and remained unchanged (P = 0.86) until day 150. At calving, no effects of breed (P = 0.34) were observed for liver Cu concentration of cows; however, Brahman calves tended (P = 0.09) to have greater liver Cu concentration than Angus calves. For Se liver concentration at calving, Angus cows tended (P = 0.07) to have greater liver Se concentration than Brahman cows; however, no breed differences (P = 0.70) were observed for liver Se concentration of calves at birth. In summary, substantial differences in multiple indicators of Cu and Se status were observed between Angus and Brahman cattle, implying that Angus and Brahman cattle possibly have different mechanisms to maintain adequate Cu and Se status.

Implementation of a standardized oral screening tool by paediatric cardiologists.

BACKGROUND: An examination of invasive procedure cancellations found that the lack of pre-procedural oral screening was a preventable cause, for children with congenital heart disease. The purpose of this study was to implement an oral screening tool within the paediatric cardiology clinic, with referral to paediatric dental providers for positive screens. The target population were children aged ≥6 months to <18 years old, being referred for cardiac procedures. METHODS: The quality implementation framework method was used for this study design. The multi-modal intervention included education, audit and feedback, screening guidelines, environmental support, and interdisciplinary collaboration. Baseline rates for oral screenings were determined by retrospective chart audit from January 2018 to January 2019 (n = 211). Provider adherence to the oral screening tool was the outcome measure. Positive oral screens, resulting in referral to the paediatric dental clinic, were measured as a secondary outcome. Provider adherence rates were used as a process measure. RESULTS: Data collected over 14 weeks showed a 29% increase in documentation of oral screenings prior to referral, as compared to the retrospective chart audit. During the study period, 13% of completed screenings were positive (n = 5). Provider compliance for the period was averaged at 70% adherence. CONCLUSION: A substantial increase in pre-procedural oral screenings by paediatric cardiologists was achieved using the quality implementation framework and targeted interventions.

Maternal supplementation of energy and protein, but not methionine hydroxy analog, enhanced postnatal growth and response to vaccination in Bos indicus-influenced beef offspring.

A 2-yr study evaluated the growth and postvaccination immune response of beef calves born from heifers offered no supplementation or pre- and postpartum supplementation of sugarcane molasses + urea with or without methionine hydroxy analog (MHA). On day 0 of each year (57 ± 5 d prepartum), Brangus crossbred beef heifers (n = 36/yr; 20 to 22 mo of age) were stratified by their initial body weight (BW; 396 ± 24.1 kg) and body condition score (BCS; 5.6 ± 0.43) and randomly allocated into 1 of 12 bahiagrass (Paspalum notatum) pastures (3 heifers/pasture). Treatments were randomly assigned to pastures (4 pastures/treatment/yr) and consisted of no supplementation (NOSUP) and supplementation of sugarcane molasses + urea (7.2 kg of DM/heifer/wk) with (MOL+) or without (MOL-) fortification with 105 g/heifer/wk of MHA. Treatments were provided from 57 ± 5 d prepartum until 17 ± 5 d postpartum (day 0 to 74). On day 74, all heifer-calf pairs were combined and managed as a single group until the end of the breeding season (day 237). Calves were early weaned at 89 ± 5 d of age (day 147), limit-fed at 3.5% of BW (DM basis) in drylot until day 201, and vaccinated against respiratory disease pathogens on days 160 and 188. Prepartum BCS on day 44 did not differ (P = 0.26) between MOL+ and MOL- heifers but both groups had greater (P < 0.0001) BCS than NOSUP heifers. Plasma concentrations of l-methionine on day 44 were the greatest (P ≤ 0.04) for MOL+ heifers and did not differ (P = 0.40) between NOSUP vs. MOL- heifers. Calf birth BW did not differ (P = 0.13) among treatments. Calf average daily gain (ADG) from birth to day 201 did not differ (P ≥ 0.17) between MOL+ vs. MOL- calves, but both groups had greater (P ≤ 0.05) ADG from birth to day 201 than NOSUP calves. Calf postvaccination plasma concentrations of glucose, cortisol, and haptoglobin did not differ among treatments (P ≥ 0.13). However, plasma concentrations of IGF-1 on day 167 and the overall positive vaccine seroconversion did not differ (P ≥ 0.18) between MOL- and MOL+ calves, but both were greater (P ≤ 0.04) compared with NOSUP calves. Hence, maternal supplementation of sugarcane molasses + urea increased BCS at calving and offspring BW gain and response to vaccination against respiratory pathogens compared with no maternal supplementation. MHA inclusion into maternal supplements effectively increased maternal plasma l-methionine concentrations but did not enhance maternal BCS at calving and offspring growth and postvaccination immune response.

Use of grape residue flour in lactating dairy sheep in heat stress: Effects on health, milk production and quality.

The objective of this study was to evaluate the effects of grape residue flour (GRF) on antioxidant activities, biochemistry variables, components of the immune system and milk production and quality of Lacaune sheep in heat stress. Twenty-seven multiparous lactating sheep [50 ±â€¯1.8 days (d) milking] were stratified by initial body weight, age, date of lambing and milk production and assigned randomly to 1 of 3 treatments (9 sheep/treatment): no GRF supplementation (control group) or supplementation at 1% (10 g/kg GRF) or 2% (20 g/kg GRF) of GRF (bark and seed) in the concentrate (grains and minerals mixture). Each ewe received 0.8 kg/d of concentrate, 3.6 kg/d of corn silage, and 0.25 kg/d of Cynodon spp hay. Milk production along with blood and milk samples were collected on d 1, 10 and 15. The 2% GRF sheep had increased serum concentrations of superoxide dismutase and glutathione peroxidase activity on d 15 compared to control sheep. Over time (d 10 to 15), lipid peroxidation was reduced in 2% GRF sheep. Total serum antioxidant capacity was greater in 2% GRF sheep compared to control sheep on d 10 and 15. Superoxide dismutase and glutathione peroxidase activity in milk samples were greater in 2% GRF sheep compared to control sheep. Supplementation with GRF did not affect milk production but GRF sheep were more efficient compared to control sheep. Protein and lactose concentrations were similar between treatments, but total solids and fat concentrations were greater in 2% GRF sheep compared to control sheep on d 15. Somatic cell count was reduced in GRF sheep compared to control sheep. In summary, supplementation with 2% GRF in dairy sheep in heat stress resulted in antioxidant and anti-inflammatory responses, which improved milk quality and reduced somatic cell count and lipid peroxidation.

The Lived Experience of International Graduate Nursing Students Engaged With the US Educational System.

BACKGROUND: Strategies are needed to address social and cultural challenges faced by international students pursuing education at American universities. PURPOSE: The purpose of this study was to explore the lived experiences of graduate international nursing students enrolled in a graduate nursing program. METHOD: This descriptive phenomenological study explored the experiences of twelve international graduate nursing students. RESULTS: Through the use of Colaizzi's (1978) method of qualitative data analysis, five emergent themes were identified: Experiencing Cultural Differences, Taking on the New Role, Navigating Challenges, Reaping Benefits, and Unfulfilled Wishes. Within these themes, unique ideas of 'wanting to do good for their home country', strategies students used for 'cultural adjustment', and 'unmet or conflicting expectations' were present that have implications for nurse educators and program administrators. CONCLUSIONS: Future recommendations for faculty include front loading professional writing support for students, exploring opportunities for increased intercultural interactions in and out of the classroom, and workshops to increase faculty cultural awareness. The theme of 'unfulfilled wishes' provides a rich understanding of goals that international students bring to the learning environment. Additional research studies are needed to explore the experiences of international nursing students in other fields and after they return home to their native country.

Fostering Student Success: Turning Off Practice Assessment Rationales to Improve Proctored Assessment Scores.

Preparing students with opportunities to practice critical thinking skills is essential to success on the National Council Licensure Examination. This pilot study was conducted to explore the effectiveness of turning off practice assessment answers and rationales on a standardized examination. A retrospective study compared students' scores when rationales were turned on and turned off. Eighty-one percent of the students were able to achieve the benchmark when the answers and rationales were turned off as compared to 71 percent when turned on. Faculty can utilize these findings to inform remediation strategies.

From Introduction to Integration: Providing Community-Engaged Structure for Interprofessional Education.

BACKGROUND: Training future healthcare profession students using interprofessional education (IPE) is critical to improve quality of health care and patient safety. OBJECTIVE: The objective of this study was to implement an IPE program and determine student satisfaction with each session, including a clinical case requiring teams with members from each profession addressing clinical scenarios. SUBJECTS: The subjects of this study were students from Athletic Training, Medicine, Nursing, Pharmacy, Physical Therapy, Physician Assistant, Social Work, and Speech-Language Pathology. METHODS: Evaluations, administered to all participating students, consisted of Likert-style responses, rating agreement with a series of questions, and space for descriptive comments. Score differences for each question were compared using independent group t-tests with a P-value of 0.05 to determine statistical significance. RESULTS: There were statistically higher satisfaction ratings for the problem-based learning case when compared to less interactive sessions (P < 0.0001). CONCLUSION: Students perceived benefits of the IPE program. Perceptions improved when various students had the opportunity to work together on clinically relevant problems.

Children with severe early childhood caries: streptococci genetic strains within carious and white spot lesions.

BACKGROUND AND OBJECTIVES: Mutans streptococci (MS) are one of the major microbiological determinants of dental caries. The objectives of this study are to identify distinct MS and non-MS streptococci strains that are located at carious sites and non-carious enamel surfaces in children with severe early childhood caries (S-ECC), and assess if cariogenic MS and non-cariogenic streptococci might independently exist as primary bacterial strains on distinct sites within the dentition of individual children. DESIGN: Dental plaque from children (N=20; aged 3-6) with S-ECC was collected from carious lesions (CLs), white spot lesions (WSLs) and non-carious enamel surfaces. Streptococcal isolates (N=10-20) from each site were subjected to polymerase chain reaction (PCR) to identify MS, and arbitrarily primed-PCR for assignment of genetic strains. Primary strains were identified as ≥50% of the total isolates surveyed at any site. In several cases, strains were characterized for acidurity using ATP-driven bioluminescence and subjected to PCR-determination of potential MS virulence products. Identification of non-MS was determined by 16S rRNA gene sequencing. RESULTS: Sixty-four independent MS or non-MS streptococcal strains were identified. All children contained 1-6 strains. In many patients (N=11), single primary MS strains were identified throughout the dentition. In other patients (N=4), primary MS strains were identified within CLs that were distinct from primary strains found on enamel. Streptococcus gordonii strains were identified as primary strains on enamel or WSLs in four children, and in general were less aciduric than MS strains. CONCLUSIONS: Many children with S-ECC contained only a single primary MS strain that was present in both carious and non-carious sites. In some cases, MS and non-cariogenic S. gordonii strains were found to independently exist as dominant strains at different locations within the dentition of individual children, and the aciduric potential of these strains may influence susceptibility in the development of CLs.

Mutans streptococci genetic strains in children with severe early childhood caries: follow-up study at one-year post-dental rehabilitation therapy.

BACKGROUND: Genotypic strains of cariogenic mutans streptococci (MS) may vary in important virulence properties. In previous published studies, we identified 39 MS strains from pediatric patients undergoing full-mouth dental rehabilitation, including the removal and/or repair of carious lesions and application of antimicrobial rinse and fluoride varnish. OBJECTIVES: The objectives of this current 1-year follow-up study are to assess the variability of MS strains that occur at 1-year post-rehabilitation and characterize the xylitol-resistance properties of MS strains that predominate. METHODS: Plaque from five children with severe early childhood caries was collected 1-year post-rehabilitation. MS isolates were subjected to arbitrarily primed-polymerase chain reaction (AP-PCR) for identification of genetic strains and in vitro xylitol-inhibition experiments. To more precisely define strain distributions within each patient, we isolated large numbers of isolates per patient. RESULTS: MS strains diminished from several strains pre-rehabilitation, to one dominant strain at 1-year post-rehabilitation, with several new emergent strains. The majority of the clinical MS strains, as well as the Streptococcus mutans laboratory strains ATCC 25175 and 35668, were predicted to undergo 50% inhibition with 2.48-5.58% xylitol, with some clinical MS strains being significantly more resistant in vitro. CONCLUSIONS: Our follow-up study using patients from the original cohort demonstrates that specific MS strains are dominant at 1-year post-dental rehabilitation. Most of the clinical MS strains are similar in xylitol resistance to the attenuated S. mutans ATCC control strains, with some strains being more resistant to xylitol in vitro.

Children with severe early childhood caries: pilot study examining mutans streptococci genotypic strains after full-mouth caries restorative therapy.

PURPOSE: Genotypic strains of mutans streptococci (MS) may vary in important virulence properties and be differentially affected by specific components of full-mouth caries restorative therapy. The purpose of this pilot study was to identify mutans streptococci strains that predominate following caries restorative therapy. METHODS: Plaque from 7 children with severe early childhood caries was collected before and following therapy. MS isolates (N=828) were subjected to polymerase chain reaction (PCR) and arbitrarily primed-PCR (AP-PCR) for assignment within MS strains. Determining the longitudinal changes in MS strain distribution over time within each patient required the isolation of larger numbers of isolates per patient, but from fewer patients. RESULTS: Up to 39 genotypic strains of Streptococcus mutans and Streptococcus sobrinus, and 7 genotypic strains of non-MS streptococci were identified by AP-PCR and 16S ribosomal rRNA gene sequencing. The number of MS strains isolated from each patient were 3 to 7 prior to treatment, diminishing to 1 to 2 dominant MS strains in most patients 6 months following therapy. CONCLUSIONS: Caries restorative therapy resulted in shifts of specific mutans streptococcus and non-mutans streptococcus strains. The implications are that caries restorative therapy affects the distribution of MS strains, and that well-accepted practices for caries prevention should be more closely examined for efficacy.