Characterization of endophytic bacteriome diversity and associated beneficial bacteria inhabiting a macrophyte Eichhornia crassipes.

Introduction: The endosphere of a plant is an interface containing a thriving community of endobacteria that can affect plant growth and potential for bioremediation. Eichhornia crassipes is an aquatic macrophyte, adapted to estuarine and freshwater ecosystems, which harbors a diverse bacterial community. Despite this, we currently lack a predictive understanding of how E. crassipes taxonomically structure the endobacterial community assemblies across distinct habitats (root, stem, and leaf). Methods: In the present study, we assessed the endophytic bacteriome from different compartments using 16S rRNA gene sequencing analysis and verified the in vitro plant beneficial potential of isolated bacterial endophytes of E. crassipes. Results and discussion: Plant compartments displayed a significant impact on the endobacterial community structures. Stem and leaf tissues were more selective, and the community exhibited a lower richness and diversity than root tissue. The taxonomic analysis of operational taxonomic units (OTUs) showed that the major phyla belonged to Proteobacteria and Actinobacteriota (> 80% in total). The most abundant genera in the sampled endosphere was Delftia in both stem and leaf samples. Members of the family Rhizobiaceae, such as in both stem and leaf samples. Members of the family Rhizobiaceae, such as Allorhizobium- Neorhizobium-Pararhizobium-Rhizobium were mainly associated with leaf tissue, whereas the genera Nannocystis and Nitrospira from the families Nannocystaceae and Nitrospiraceae, respectively, were statistically significantly associated with root tissue. Piscinibacter and Steroidobacter were putative keystone taxa of stem tissue. Most of the endophytic bacteria isolated from E. crassipes showed in vitro plant beneficial effects known to stimulate plant growth and induce plant resistance to stresses. This study provides new insights into the distribution and interaction of endobacteria across different compartments of E. crassipes Future study of endobacterial communities, using both culture-dependent and -independent techniques, will explore the mechanisms underlying the wide-spread adaptability of E. crassipesto various ecosystems and contribute to the development of efficient bacterial consortia for bioremediation and plant growth promotion.

Sub-optimal nutrient regime coupled with Bacillus and Pseudomonas sp. inoculation influences trichome density and cannabinoid profiles in drug-type Cannabis sativa.

Cannabis sativa remains under heavy legal restriction around the globe that prevents extensive investigations into agricultural applications for improving its development. This work investigates the potential of specific plant growth-promoting rhizobacteria (PGPR) to improve Cannabis cannabinoid yield through increased trichome densities on floral organs, and to determine if sub-optimal environmental conditions would affect the outcomes of PGPR presence by altering plant development and cannabinoid profiles. Here, Pseudomonas sp. or Bacillus sp. were applied to the root system either separately or in a consortium to determine the effect of this bacterial treatment on the density of stalked glandular trichomes. Further, a low nutrient regime was applied for the first half of plant development to determine if an environmental stressor interacts with the effects of the microbial treatments on stalked trichome densities. Following 8 weeks of flower development, trichome density on calyces and bracts of inflorescences were determined using microscopy. Our findings unexpectedly indicate that recommended nutrient levels were linked to a decreasing trend in trichome densities with PGPR inoculations, but a low nutrient regime coupled with PGPR treatment increased them. Cannabinoid content is partially consistent with these results, in that a low nutrient regime increased the abundance of key cannabinoids compared to recommended regimes, with Bacillus sp. inoculation linked to the greatest number of significant changes between the two nutrient regimes. Overall, this work provides insight into how PGPR presence affects Cannabis stalked trichome development and cannabinoid profiles, and how environmental stressors can affect, and even enhance, trichome densities and influence major cannabinoid production, thereby pointing towards avenues for reducing the reliance on synthetic fertilizers during plant production without compromising yield.

Inoculum dose-disease response relationships for the pea root rot pathogen, Aphanomyces euteiches, are dependent on soil type and other pathogens.

The oomycete pathogen, Aphanomyces euteiches, was implicated for the first time in pea and lentil root rot in Saskatchewan and Alberta in 2012 and 2013. Subsequent surveys from 2014 to 2017 revealed that Aphanomyces root rot (ARR) was widespread across the Canadian prairies. The absence of effective chemical, biological, and cultural controls and lack of genetic resistance leave only one management option: avoidance. The objectives of this study were to relate oospore levels in autoclaved and non-autoclaved soils to ARR severity across soil types from the vast prairie landscape and to determine the relationship of measured DNA quantity of A. euteiches using droplet digital PCR or quantitative PCR to the initial oospore inoculum dose in soils. These objectives support a future end goal of creating a rapid assessment method capable of categorizing root rot risk in field soil samples to aid producers with pulse crop field selection decisions. The ARR severity to oospore dose relationship was statistically significantly affected by the soil type and location from which soils were collected and did not show a linear relationship. For most soil types, ARR did not develop at oospore levels below 100/g soil, but severity rose above this level, confirming a threshold level of 100 oospores/g soil for disease development. For most soil types, ARR severity was significantly higher in non-autoclaved compared to autoclaved treatments, demonstrating the role that other pathogens play in increasing disease severity. There was a significant linear relationship between DNA concentrations measured in soil and oospore inoculum concentration, although the strength of the relationship was better for some soil types, and in some soil types, DNA measurement results underestimated the number of oospores. This research is important for developing a root rot risk assessment system for the Canadian prairies based on soil inoculum quantification, following field validation of soil quantification and relationship to root rot disease severity.

A Single Intranasal Dose of Bacterial Therapeutics to Calves Confers Longitudinal Modulation of the Nasopharyngeal Microbiota: a Pilot Study.

To address the emergence of antimicrobial-resistant pathogens in livestock, microbiome-based strategies are increasingly being sought to reduce antimicrobial use. Here, we describe the effects of intranasal application of bacterial therapeutics (BTs) on the bovine respiratory microbiota and used structural equation modeling to investigate the causal networks after BT application. Beef cattle received (i) an intranasal cocktail of previously characterized BT strains, (ii) an injection of metaphylactic antimicrobial (tulathromycin), or (iii) intranasal saline. Despite being transient colonizers, inoculated BT strains induced longitudinal modulation of the nasopharyngeal bacterial microbiota while showing no adverse effect on animal health. The BT-mediated changes in bacteria included reduced diversity and richness and strengthened cooperative and competitive interactions. In contrast, tulathromycin increased bacterial diversity and antibiotic resistance and disrupted bacterial interactions. Overall, a single intranasal dose of BTs can modulate the bovine respiratory microbiota, highlighting that microbiome-based strategies have potential in being utilized to mitigate bovine respiratory disease in feedlot cattle. IMPORTANCE Bovine respiratory disease (BRD) remains the most significant health challenge affecting the North American beef cattle industry and results in $3 billion in economic losses yearly. Current BRD control strategies mainly rely on antibiotics, with metaphylaxis commonly employed to mitigate BRD incidence in commercial feedlots. However, the emergence of multidrug-resistant BRD pathogens threatens to reduce the efficacy of antimicrobials. Here, we investigated the potential use of novel bacterial therapeutics (BTs) to modulate the nasopharyngeal microbiota in beef calves, which are commonly administered metaphylactic antibiotics to mitigate BRD when sourced from auction markets. By direct comparison of the BTs with an antibiotic commonly used for BRD metaphylaxis in feedlots, this study conveyed the potential use of the BTs to modulate respiratory microbiome and thereby improve resistance against BRD in feedlot cattle.

Effect of rest, post-rest transport duration, and conditioning on performance, behavioural, and physiological welfare indicators of beef calves.

The aim of this study was to assess the effects of conditioning, rest, and post-rest transport duration on welfare indicators of 6-7 mo old beef calves following a 20-h transport. Three hundred and twenty-eight weaned calves (237 ± 29.7 kg of BW) were randomly assigned to a 2 × 2 × 2 nested factorial design: conditioning, conditioned (C) or non-conditioned (N); rest, 0 (R0) or 8 (R8) h, and post-rest transport, 4 (T4) or 15 (T15) h. Calves were sampled before (LO1) and after (UN1) the initial 20-h journey, before (LO2) and after (UN2) the additional 4 or 15-h journey, and at 1, 2, 3, 5, 14, and 28 d after UN2. Data was analyzed using the GLIMMIX procedure of SAS. Fixed effects included conditioning, transport, and time nested within rest period, while random effects included animal and pen. Greater shrink (p < 0.01) was observed in C than N calves after the initial 20-h transport. During the first week after transportation, the mean ADG of N calves was greater than C calves (p < 0.01). From d 14 to d 28, however, the mean ADG of C calves was greater than N calves (p < 0.01). Flight speed, cortisol and L-lactate concentrations were greater (p ≤ 0.05) in C than N calves between LO1 and d 5, while greater (p ≤ 0.02) non-esterified fatty acids, creatine kinase, serum amyloid-A, and haptoglobin concentrations were observed in N than C calves between LO1 and d 3. The R8-T4 calves had greater (p < 0.01) ADG than R8-T15 calves between LO1 and d 5. The R0-T4 calves had greater L-lactate concentrations than R0-T15 and R8-T4 calves (both p = 0.02) on d 1. The R0 calves had greater (p < 0.01) ADG than R8 calves between 14 and 28 d. This study suggests that C calves are better fit for transport than N calves as evidenced by behavioural and physiological parameters. Fewer and inconsistent differences were observed for rest and post-rest transport treatments.

Aragonite toothpaste for management of dental calculus: A double-blinded randomized controlled clinical trial.

OBJECTIVES: Aragonite from animal origin such as cuttlefish bone powder is an abrasive with hardness properties ideal for calculus removal. The purpose of this randomized controlled trial was to test the efficacy of a cuttlebone-derived aragonite toothpaste in removing dental calculus. MATERIALS AND METHODS: Eighty-one patients who fulfilled the inclusion criteria were blindly and randomly assigned into two study groups. The intervention treatment group (n = 40) received cuttlebone toothpaste (Dr. D-Tart) and the control group (n = 41) received an off-the-shelf commercial toothpaste (Crest). Evaluations were performed before and after scaling and polishing procedures done at 3 months in order to evaluate the toothpaste's ability to remove calculus and to prevent calculus formation. Calculus, stains, plaque, and gingival indices scores, and patient satisfaction surveys were compared at baseline (first visit), 3, and 9 months, using generalized linear models and Wald's χ2 test. RESULTS: At the end of the 3-month period, the intervention group showed a 30% reduction in total calculus compared to the baseline score (p = .0006) and 45% less total calculus compared to the control group (p = .0001). Six months after scaling, the mean calculus score for Crest users was 42% higher than that for Dr. D-Tart users (p = .0692). There was a significant improvement in the gingival health of cuttlebone toothpaste users at the observed intervals, and both kinds of toothpaste achieved comparable results in terms of plaque and stains removal. CONCLUSIONS: Aragonite toothpaste can remove calculus, prevent calculus formation, and improve gingival health. Patients are generally satisfied with the performance of the aragonite toothpaste. CLINICAL SIGNIFICANCE: Animal-derived aragonite toothpaste (Dr. D-Tart) shows promising efficacy in removing calculus, preventing calculus formation, and for the improvement of gingival health. CLINICAL TRIAL ID: A08-M35-16B.

Influence of Production Factors on Beef Primal Tissue Composition.

This study used 1076 crossbred steers to evaluate the effects of calf-fed and yearling-fed beef production systems, implant strategies (with and without implants), and their interactions on the primal tissue composition (lean and fat components) of individual primal cuts using complete carcass dissection data. The results indicate that production system × implant interactions affected loin and rib primal weight percentages as well as marbling (p < 0.05) but did not affect the dissectible lean and fat contents of the individual primal cut (p > 0.05). Implants increased lean and decreased fat tissue contents of primal cut; however, the production system only affected lean content in the loin (p < 0.05) and fat content in the loin, round, and rib (p < 0.05). Redundancy analysis revealed a strong association between Angus breed percentage and marbling, as well as between Simmental breed percentage and multiple primal lean traits. Response surface regression models explained less variability in the tissue composition traits in calf-fed compared with yearling-fed animals, suggesting the need for further exploration using genomic studies.

Marginal quality water arbitrated essential oil contents in metal hoarded flower petals of scented roses.

Climate change and the consequent alteration in agricultural circumstances enhance the susceptibility of fresh water use particularly in water-scarce regions. Marginal quality water reuse is a common alternative practice but possible perils of metal accretion in plant parts are mostly ignored. The present research aimed to probe the impact of treated wastewater (TWW) and untreated wastewater (UTWW) on metal accumulation in flower petals and their influence on essential oil contents of fragrant Rosa species (R. Gruss-an-teplitz, R. bourboniana, R. centifolia, R. damascena) in a peri-urban area of Faisalabad, Pakistan during January, 2017 to December, 2018. The mineral and chemical contents in canal water (CW) and TWW were less than recommended levels of national environmental quality standards (NEQS) for wastewater of Pakistan. The experimentally UTWW possessed higher electrical conductivity (EC), biological and chemical oxygen demand (BOD and COD), and some metals (Pb, Co, Cr) that were above the permissible levels. The experimental data revealed that except Cr other metals contents in the flower petals were less than the WHO recommended limits (for medicinal plants) under experimental irrigation regimes. Rosa centifolia and R. damascena possessed higher metal i.e. Zn, Cu, Pb, Cr, Co contents while Fe and Ni contents were higher in R. Gruss-an-Teplitz and R. bourboniana respectively. There were twelve constituents which were detected in essential oil by gas chromatography. Major constituents were phenyl ethyl alcohol, citronellol, geranyl acetate, γ- undelactone, methyl eugenol, and limonene whose share was 48.17%, 41.11%, 8.46%, 4.82%, 4.44%, and 4.15% respectively whereas concentrations of other 06 constituents were less than 3.7%. Phenyl ethyl alcohol, lion shared constituent of essential oil was found highest (48.17%) in R. Gruss-an-Teplitz whereas minimum level was recorded in R. damascena (28.84%) under CW. In contrast, citronellol (chief component of fragrance) was highest in R. damascena (41.11%) in UTWW while the lowest level was found in R. Gruss-an-Teplitz (17.41%) in CW. This study confirmed the variations in metal concentrations of Rosa species due to different absorbability of each metal in flower petals. It also indicates that wastewater did not affect the composition but there were quantitative differences in aroma constituents and chemical composition of essential oil.

The Nasopharyngeal, Ruminal, and Vaginal Microbiota and the Core Taxa Shared across These Microbiomes in Virgin Yearling Heifers Exposed to Divergent In Utero Nutrition during Their First Trimester of Gestation and in Pregnant Beef Heifers in Response to Mineral Supplementation.

In the present study, we evaluated whether the nasopharyngeal, ruminal, and vaginal microbiota would diverge (1) in virgin yearling beef heifers (9 months old) due to the maternal restricted gain during the first trimester of gestation; and (2) in pregnant beef heifers in response to the vitamin and mineral (VTM) supplementation during the first 6 months of pregnancy. As a secondary objective, using the microbiota data obtained from these two cohorts of beef heifers managed at the same location and sampled at the same time, we performed a holistic assessment of the microbial ecology residing within the respiratory, gastrointestinal, and reproductive tract of cattle. Our 16S rRNA gene sequencing results revealed that both α and ß-diversity of the nasopharyngeal, ruminal and vaginal microbiota did not differ between virgin heifers raised from dams exposed to either a low gain (targeted average daily gain of 0.28 kg/d, n = 22) or a moderate gain treatment (0.79 kg/d, n = 23) during the first 84 days of gestation. Only in the vaginal microbiota were there relatively abundant genera that were affected by maternal rate of gain during early gestation. Whilst there was no significant difference in community structure and diversity in any of the three microbiota between pregnant heifers received no VTM (n = 15) and VTM supplemented (n = 17) diets, the VTM supplementation resulted in subtle compositional alterations in the nasopharyngeal and ruminal microbiota. Although the nasopharyngeal, ruminal, and vaginal microbiota were clearly distinct, a total of 41 OTUs, including methanogenic archaea, were identified as core taxa shared across the respiratory, gastrointestinal, and reproductive tracts of both virgin and pregnant heifers.

Formalin fumigation and steaming of various composts differentially influence the nutrient release, growth and yield of muskmelon (Cucumis melo L.).

Nutrient disorder and presence of disease-causing agents in soilless media negatively influence the growth of muskmelon. To combat these issues, use of environmentally-friendly sanitation techniques is crucial for increased crop productivity. The study was conducted under greenhouse and field conditions to investigate the effect of two different sanitation techniques: steaming and formalin fumigation on various media's characteristics and their impact on muskmelon yield. Media: jantar, guar, wheat straw and rice hull and peat moss of 10% air-filled porosity and sanitized with formalin and steaming. Steaming of guar, jantar, and wheat straw increased the phosphorus (P) and potassium (K) concentrations by 13.80-14.86% and 6.22-8.45% over formalin fumigation. Likewise, P and K concentrations in muskmelon were higher under steaming. Steaming significantly inhibited the survival of Fusarium wilt sp. melonis, root knot nematode sp. meloidogyne and nitrifying bacteria in media than formalin fumigation. In conclusion, steaming decreased the prevalence of nitrifying bacteria and pathogens which thus improved the NO3--N:NH4+-N ratios, P and K nutritional balance both in the media and muskmelon transplants. Hence, steaming as an environment-friendly approach is recommended for soilless media. Further, optimization of steaming for various composts with different crops needs to be investigated with steaming teachnique.

Identification of Differential Drought Response Mechanisms in Medicago sativa subsp. sativa and falcata through Comparative Assessments at the Physiological, Biochemical, and Transcriptional Levels.

Alfalfa (Medicago sativa L.) is an extensively grown perennial forage legume, and although it is relatively drought tolerant, it consumes high amounts of water and depends upon irrigation in many regions. Given the progressive decline in water available for irrigation, as well as an escalation in climate change-related droughts, there is a critical need to develop alfalfa cultivars with improved drought resilience. M. sativa subsp. falcata is a close relative of the predominantly cultivated M. sativa subsp. sativa, and certain accessions have been demonstrated to exhibit superior performance under drought. As such, we endeavoured to carry out comparative physiological, biochemical, and transcriptomic evaluations of an as of yet unstudied drought-tolerant M. sativa subsp. falcata accession (PI 641381) and a relatively drought-susceptible M. sativa subsp. sativa cultivar (Beaver) to increase our understanding of the molecular mechanisms behind the enhanced ability of falcata to withstand water deficiency. Our findings indicate that unlike the small number of falcata genotypes assessed previously, falcata PI 641381 may exploit smaller, thicker leaves, as well as an increase in the baseline transcriptional levels of genes encoding particular transcription factors, protective proteins, and enzymes involved in the biosynthesis of stress-related compounds. These findings imply that different falcata accessions/genotypes may employ distinct drought response mechanisms, and the study provides a suite of candidate genes to facilitate the breeding of alfalfa with enhanced drought resilience in the future.

Missing and unerupted teeth in osteogenesis imperfecta.

INTRODUCTION: Osteogenesis imperfecta (OI) is a genetic disorder characterized by bone fragility and craniofacial and dental abnormalities such as congenitally missing teeth and teeth that failed to erupt which are believed to be doubled in OI patients than normal populations and were associated with low oral health quality of life. However, the etiology of these abnormalities remains unclear. To understand the factors influencing missing and unerupted teeth, we investigated their prevalence in a cohort of OI patients as a function of the clinical phenotype (OI type), the genetic variant type, the tooth type and the onset of bisphosphonate treatment. METHOD: A total of 144 OI patients were recruited from The Shriners Hospital, Montreal, Canada, between 2016 and 2017. Patients were evaluated using intraoral photographs and panoramic radiographs. Missing teeth were evaluated in all patients, and unerupted teeth were assessed only in patients ≥15 years old (n = 82). RESULTS: On average, each OI patient had 2.4 missing teeth and 0.8 unerupted teeth, and the most common missing and unerupted teeth were the premolars and the upper second molars, respectively. These phenomena were more prominent in OI type III and IV than in OI type I, and were not sex or age-related. Missing teeth were significantly more common in patients with C-propeptide variants than all other variants (p-value <0.05). Unerupted teeth were significantly more common in patients with α1 and α2 glycine variants or substitutions than in those with haploinsufficiency variants. Early-onset of bisphosphonate treatment would significantly increase the risk of unerupted teeth in patients with OI types III and IV (OR = 1.68, 95% CI (1.15-1.53)). CONCLUSION: The prevalence of missing and unerupted teeth at the tooth type level in OI patients varies according to the nature of the collagen variants and the OI type. These findings highlight the role of collagen in tooth development and eruption.

Correlation between L-Lactate Concentrations in Beef Cattle, Obtained Using a Hand-Held Lactate Analyzer and a Lactate Assay Colorimetric Kit.

Lactate is a product of anaerobic glycolysis, used in animal research as an indicator of muscle fatigue. Therefore, it has been used as an indicator of cattle response to long distance transportation. The aim of this study was to assess the relationship of L-lactate concentrations measured using a Lactate Scout+ analyzer and a traditional lactate assay colorimetric kit. Blood samples were collected by venipuncture from 96 steers (Black or Red Angus × Hereford/Simmental and Black or Red Angus × Charolais; 247 ± 38.2 kg BW) prior to loading (LO1) and after 36 h of transport, and prior to reloading and after an additional 4 h of road transportation, and on d 1, 2, 3, 5, 14, and 28 after transport. The Lactate Scout+ analyzer strip was dipped in blood at the time of sampling, while blood samples were collected into sodium fluoride tubes for use in the colorimetric analysis. Pearson correlations were calculated to assess the strength of the relationship between the experimental methods for the quantification of L-lactate concentrations. The magnitude and direction of the correlation, and the level of statistical significance varied over the observed time points, ranging from r = -0.03 (p = 0.75; LO1) to r = 0.75 (p < 0.0001; d 3). The correlation for the pooled data was weak but statistically significant (r = 0.33, p < 0.0001). Based on the low magnitude of the correlation due to variability across sampling time points in this study, the Lactate Scout+ analyzer is not a suitable alternative to a lab-based assay (considered the gold standard) for measuring L-lactate in transported cattle.

Osteogenesis imperfecta tooth level phenotype analysis: Cross-sectional study.

INTRODUCTION: Dental anomalies in Osteogenesis imperfecta (OI), such as tooth discoloration, pulp obliteration (calcified dental pulp space), and taurodontism (enlarged dental pulp space) vary between and within patients. To better understand the associations and variations in these anomalies, a cross-sectional study was designed to analyze the dental phenotype in OI patients at the individual tooth type. METHOD: A cohort of 171 individuals with OI type I, III and IV, aged 3-55 years, were recruited and evaluated for tooth discoloration, pulp obliteration, and taurodontism at the individual tooth level, using intraoral photographs and panoramic radiographs. RESULTS: Genetic variants were identified in 154 of the participants. Patients with Helical α1 and α2 glycine substitutions presented the highest prevalence of tooth discoloration, while those with α1 Haploinsufficiency had the lowest (<10%). C-propeptide variants did not cause discoloration but resulted in the highest pulp obliteration prevalence (~%20). The prevalence of tooth discoloration and pulp obliteration was higher in OI types III and IV and increased with age. Tooth discoloration was mainly observed in teeth known to have thinner enamel (i.e. lower anterior), while pulp obliteration was most prevalent in the first molars. A significant association was observed between pulp obliteration and tooth discoloration, and both were associated with a lack of occlusal contact. Taurodontism was only found in permanent teeth and affected mostly first molars, and its prevalence decreased with age. CONCLUSION: The dental phenotype evaluation at the tooth level revealed that different genetic variants and associated clinical phenotypes affect each tooth type differently, and genetic variants are better predictors of the dental phenotype than the type of OI. Our results also suggest that tooth discoloration is most likely an optical phenomenon inversely proportional to enamel thickness, and highly associated with pulp obliteration. In turn, pulp obliteration is proportional to patient age, it is associated with malocclusion and likely related to immature progressive dentin deposition. Taurodontism is an isolated phenomenon that is probably associated with delayed pulpal maturation.

Distribution of Pest Wireworm (Coleoptera: Elateridae) Species in Alberta, Saskatchewan, and Manitoba (Canada).

We present findings of a general survey of pest wireworms in Alberta, Saskatchewan, and Manitoba conducted from 2004 to 2019; the first such survey of the Canadian Prairie provinces since that published by Glen et al. (1943). Samples were collected from 571 farmland locations where crop damage from wireworms was observed or suspected, and a total of 5,704 specimens (3,548 larvae, 2,156 beetles) were identified. Most specimens (96.9%) were identified as Hypnoidus bicolor (3,278), Selatosomus aeripennis destructor (1,280), Limonius californicus (842), and Aeolus mellillus (125). This suggests that H. bicolor has replaced S. a. destructor as the predominant species and that the relative importance of L. californicus as a pest species has increased since earlier reports. Despite the relatively small number of specimens collected per location (approx. 10), H. bicolor and S. a. destructor, and S. a. destructor and L. californicus were frequently collected at or near the same location (within 1 km). We provide species records and incidence of co-occurrence at different spatial scales, discuss potential reasons for and implications of shifts in species composition, implications of species co-occurrence for managing wireworm pests in crop production, and outline pertinent research needs. A path analysis approach used to correlate incidence of the main species with various soil characteristics indicated that organic matter, cation exchange capacity, and water retention capacity all had a significant species-specific influence on wireworm presence.

Effects of conditioning, source, and rest on indicators of stress in beef cattle transported by road.

Consumers are increasingly concerned about the welfare of farm animals, especially during transportation. The Canadian transport regulations state that weaned cattle require an 8 h rest after 36 h of transport. There are, however, only a few studies that assess the effect of rest on the welfare of beef cattle. The aim of this study was to assess the effect of conditioning, source and rest on indicators of welfare in 7-8-month-old beef calves during long-distance transport. Treatments consisted of a 2 × 2 × 2 factorial design where the main factors were conditioning: conditioned (C; n = 160) or non-conditioned (N; n = 160); source: auction market (A; n = 160) or ranch direct (R; n = 160); and rest: 0 h (0 h; n = 160) or 8 h (8 h; n = 160). Means of non-esterified fatty acids (NEFA), serum amyloid-A, haptoglobin, creatine kinase (CK), and percentage of time standing from N calves were greater than C calves (all p ≤ 0.05). Means of percentage of time standing and CK of R calves were greater than A calves (both p ≤ 0.05). The mean of NEFA of 0 h calves was greater than the 8 h calves (p < 0.01), while the percentage of time standing of 0 h calves was less than 8 h calves (p < 0.01). Statistically significant differences between means of NEFA and standing percentage, were observed between 0 h and 8h calves. Few and inconsistent indicators of reduced welfare were observed between auction market and ranch direct calves, while non-conditioning was associated with greater physiological and behavioural indicators of reduced welfare. Based on these results, conditioning should be implemented as a management practice to improve the welfare of transported calves.

Quantifying fluorescent glycan uptake to elucidate strain-level variability in foraging behaviors of rumen bacteria.

Gut microbiomes, such as the microbial community that colonizes the rumen, have vast catabolic potential and play a vital role in host health and nutrition. By expanding our understanding of metabolic pathways in these ecosystems, we will garner foundational information for manipulating microbiome structure and function to influence host physiology. Currently, our knowledge of metabolic pathways relies heavily on inferences derived from metagenomics or culturing bacteria in vitro. However, novel approaches targeting specific cell physiologies can illuminate the functional potential encoded within microbial (meta)genomes to provide accurate assessments of metabolic abilities. Using fluorescently labeled polysaccharides, we visualized carbohydrate metabolism performed by single bacterial cells in a complex rumen sample, enabling a rapid assessment of their metabolic phenotype. Specifically, we identified bovine-adapted strains of Bacteroides thetaiotaomicron that metabolized yeast mannan in the rumen microbiome ex vivo and discerned the mechanistic differences between two distinct carbohydrate foraging behaviors, referred to as "medium grower" and "high grower." Using comparative whole-genome sequencing, RNA-seq, and carbohydrate-active enzyme fingerprinting, we could elucidate the strain-level variability in carbohydrate utilization systems of the two foraging behaviors to help predict individual strategies of nutrient acquisition. Here, we present a multi-faceted study using complimentary next-generation physiology and "omics" approaches to characterize microbial adaptation to a prebiotic in the rumen ecosystem. Video abstract.

Intranasal Bacterial Therapeutics Reduce Colonization by the Respiratory Pathogen Mannheimia haemolytica in Dairy Calves.

Six Lactobacillus strains originating from the nasopharyngeal microbiota of cattle were previously characterized in vitro and identified as candidate bacterial therapeutics (BTs) for mitigating the bovine respiratory pathogen Mannheimia haemolytica In the present study, these BT strains were evaluated for their potential to (i) reduce nasal colonization by M. haemolytica, (ii) modulate the nasal microbiota, and (iii) stimulate an immune response in calves experimentally challenged with M. haemolytica. Twenty-four Holstein bull calves (1 to 3 weeks old) received either an intranasal BT cocktail containing 6 Lactobacillus strains (3 × 109 CFU per strain; BT + Mh group) 24 h prior to intranasal M. haemolytica challenge (3 × 108 CFU) or no BTs prior to challenge (Mh, control group). Nasal swab, blood, and transtracheal aspiration samples were collected over the course of 16 days after BT inoculation. Counts of M. haemolytica were determined by culturing, and the nasal and tracheal microbiotas were evaluated using 16S rRNA gene sequencing. Serum cytokines (interleukin-6 [IL-6], IL-8, and IL-10) were quantified by enzyme-linked immunosorbent assay (ELISA). Administration of BT reduced nasal colonization by M. haemolytica (P = 0.02), modified the composition and diversity of the nasal microbiota, and altered interbacterial relationships among the 10 most relatively abundant genera. The BT + Mh calves also had a lower relative abundance of Mannheimia in the trachea (P < 0.01) but similar cytokine levels as Mh calves. This study demonstrated that intranasal BTs developed from the bovine nasopharyngeal Lactobacillus spp. were effective in reducing nasal colonization by M. haemolytica in dairy calves.IMPORTANCE Bovine respiratory disease (BRD) is one of the significant challenges for the modern dairy industry in North America, accounting for 23 to 47% of the total mortality among pre- and postweaned dairy heifers. Mass medication with antibiotics is a common practice to control BRD in dairy cattle. However, the emergence of multidrug-resistant BRD pathogens highlights the importance of developing alternatives to antibiotics for BRD mitigation. Using a targeted approach, we recently identified 6 Lactobacillus strains originating from the bovine respiratory microbiota as candidates to be used as bacterial therapeutics (BTs) for the mitigation of the BRD pathogen Mannheimia haemolytica Here, we demonstrated that intranasal inoculation of the BT strains reduced nasal colonization by M. haemolytica in dairy calves experimentally challenged with this pathogen. This study, for the first time, shows the potential use of intranasal BTs as an alternative to mitigate BRD pathogens in cattle.

Effect of transport and rest stop duration on the welfare of conditioned cattle transported by road.

The effects of providing rest on physiological and behavioural indicators of welfare of cattle being transported by road has not been well studied in North America. New revisions to Canada's Health of Animals Regulations Part XII: Transportation of Animals indicate un-weaned and weaned calves can be transported a maximum of 12 and 36 h, respectively, before an 8 h rest is required. Therefore, the aim of this study was to assess the effects of rest duration, after 12 and 36 h of transport, on physiological and behavioural indicators of welfare in 7-8 mo-old beef calves. Three hundred and twenty conditioned calves (258 ± 23.9 kg BW) were randomly assigned to a 2 × 4 factorial design where the main factors included transport duration: 12 h (12; n = 160) and 36 h (36; n = 160) and rest stop duration: 0 h (R0; n = 80), 4 h (R4; n = 80), 8 h (R8; n = 80) and 12 h (R12; n = 80). After the resting period, animals were transported for an additional 4 h. Blood and hair samples were taken from 12 animals per treatment prior to and after the first and the 4 h transport; and then 7 h, 2 d and 28 d after the 4 h transport. The concentrations of haptoglobin, creatine kinase, non-esterified fatty acids (NEFA), lactate, and serum and hair cortisol were determined. Standing and lying behaviour was assessed for 14 d after transport, while feeding behaviour of calves in one pen per treatment were assessed for 28 d after transportation using an electronic feed bunk monitoring system. Body weight (BW), average daily gain (ADG) and shrink (%) was assessed for all calves. The data was modeled using generalized linear mixed methods (SAS PROC GLIMMIX), where transport and time (nested in rest) were considered fixed effects and animal and pen were considered random effects. Statistically significant (p < 0.05) effects of transport were observed on BW and shrink, where 36 h-transported calves had lower (p < 0.01) BW and greater (p < 0.01) shrink than 12 h-transported calves. A transport × time (nested in rest) interaction (p < 0.01) was observed for lying percentage where, 36-R8 calves had greater (p < 0.01) lying percentage than 12-R8 calves on d 1 after transportation. The area under the curve (AUC) for NEFA was greater (p < 0.01) for 36-R0 calves than 12-R0, 36-R4, and 36-R8 calves, and greater (p < 0.01) in 36-R12 calves than 12-R12 calves. Haptoglobin AUC was greater (p = 0.05) in 36-R12 than 12-R12 calves. Overall, physiological indicators of reduced welfare were greater in calves transported for 36 than 12 h, while no clear differences were observed between rest stop groups with the exception of NEFA. Based on these results, conditioned calves benefit from shorter transport durations but there was no clear evidence that calves rested 4, 8, and 12 h following transportation experienced reduced transport related stress compared to those that were not rested (0h).

Evaluation of the Nasopharyngeal Microbiota in Beef Cattle Transported to a Feedlot, With a Focus on Lactic Acid-Producing Bacteria.

The nasopharyngeal (NP) microbiota is important in defining respiratory health in feedlot cattle, with certain NP commensals potentially protecting against bovine respiratory disease (BRD) pathogens. In the present study, we evaluated longitudinal changes in the NP microbiota with a focus on lactic acid-producing bacteria (LAB) and their linkage with BRD-associated bacteria in steers (n = 13) that were first transported to an auction market, and then to a feedlot. Deep nasopharyngeal swabs were collected at the farm before transportation to the auction market (day 0), at feedlot placement (day 2), and 5 (day 7) and 12 (day 14) days after feedlot placement. Swabs were processed for the assessment of the NP microbiota using 16S rRNA gene sequencing, and for the detection of Mannheimia haemolytica, Pasteurella multocida, and Histophilus somni by culturing. Possible associations among the top 15 most relatively abundant bacterial genera were predicted using a stepwise-selected generalized linear mixed model. Correlations between LAB and BRD-associated Pasteurellaceae families were also assessed. In addition, antimicrobial activity of selected LAB isolates against M. haemolytica was evaluated in vitro. A noticeable shift was observed in the NP microbial community structure, and in the relative abundance of LAB families as a result of auction market exposure, transport and feedlot placement. Varying degrees of positive or negative associations between the 15 most relatively abundant genera were observed. Many of the LAB families were inversely correlated with the BRD-associated Pasteurellaceae family as the cattle were transported to the auction market and then to the feedlot. Nearly all steers were culture-negative for M. haemolytica and H. somni, and P. multocida became less prevalent after feedlot placement. Isolates from the Lactobacillaceae, Streptococcaceae, and Enterococcaceae families inhibited the growth of M. haemolytica. The results of this study indicated that the NP microbiota became more diverse with an increase in microbial richness following transport to an auction market and feedlot. This study provides evidence of potential cooperation and exclusion taking place in the respiratory microbial community of cattle which may be useful for developing microbial-based strategies to mitigate BRD.