Tetrameric UvrD Helicase Is Located at the E. Coli Replisome due to Frequent Replication Blocks.

DNA replication in all organisms must overcome nucleoprotein blocks to complete genome duplication. Accessory replicative helicases in Escherichia coli, Rep and UvrD, help remove these blocks and aid the re-initiation of replication. Mechanistic details of Rep function have emerged from recent live cell studies; however, the division of UvrD functions between its activities in DNA repair and role as an accessory helicase remain unclear in live cells. By integrating super-resolved single-molecule fluorescence microscopy with biochemical analysis, we find that UvrD self-associates into tetrameric assemblies and, unlike Rep, is not recruited to a specific replisome protein despite being found at approximately 80% of replication forks. Instead, its colocation with forks is likely due to the very high frequency of replication blocks composed of DNA-bound proteins, including RNA polymerase and factors involved in repairing DNA damage. Deleting rep and DNA repair factor genes mutS and uvrA, and inhibiting transcription through RNA polymerase mutation and antibiotic inhibition, indicates that the level of UvrD at the fork is dependent on UvrD's function. Our findings show that UvrD is recruited to sites of nucleoprotein blocks via different mechanisms to Rep and plays a multi-faceted role in ensuring successful DNA replication.

Evaluation of Thermal Antinociceptive Effects of Intramuscular Hydromorphone Hydrochloride in Great Horned Owls (Bubo virginianus).

Across the Americas, great horned owls (Bubo virginianus) are often presented to veterinarians for conditions requiring pain management. Although recent studies have evaluated opioid drugs in raptor species, information in Strigiformes is lacking. The objective of this study was to evaluate the analgesic effect and duration of action of hydromorphone hydrochloride, a full µ-opioid receptor agonist, in great horned owls. In a randomized, blinded, balanced crossover study, 6 adult birds (5 females and 1 male) received hydromorphone (0.3 and 0.6 mg/kg) or saline (0.9% NaCl) solution (0.03 mL/kg; control) in the left pectoral muscle, with a 7-day washout interval between treatments. Each bird was assigned an agitation-sedation score, and the thermal foot withdrawal threshold (TFWT) was measured at predetermined times before (t = 0 hours) and after treatment administration (t = 0.5, 1.5, 3, and 6 hours). Measurements of the TFWT were obtained with a test box equipped with a thermal perch, which delivered a gradually increasing temperature 40-62°C (104-143.6°F) to the right plantar surface of the owl's foot. Compared with controls, hydromorphone at 0.3 mg/kg dose resulted in significantly higher mean TFWT at 0.5 hours (P < 0.001), 1.5 hours (P = 0.003), and 3 hours (P = 0.005), whereas the 0.6 mg/kg dose resulted in significantly higher mean TFWT from 0.5 hours (P = 0.035) to 1.5 hours (P = 0.001). Both hydromorphone doses were associated with a significant change in the agitation-sedation score (P = 0.001), consistent with mild to moderate sedation. Two owls were observed tremoring after administration of the 0.6 mg/kg dose, which was not noted after the 0.5-hour timepoint; no other adverse effects were identified. This study offers scientific evidence to support the use of a µ-opioid agonist in great horned owls for pain management. Pharmacokinetics and other pharmacodynamic studies of other pain models evaluating hydromorphone and other opioid drugs in this species are still needed.

Interplay between chromosomal architecture and termination of DNA replication in bacteria.

Faithful transmission of the genome from one generation to the next is key to life in all cellular organisms. In the majority of bacteria, the genome is comprised of a single circular chromosome that is normally replicated from a single origin, though additional genetic information may be encoded within much smaller extrachromosomal elements called plasmids. By contrast, the genome of a eukaryote is distributed across multiple linear chromosomes, each of which is replicated from multiple origins. The genomes of archaeal species are circular, but are predominantly replicated from multiple origins. In all three cases, replication is bidirectional and terminates when converging replication fork complexes merge and 'fuse' as replication of the chromosomal DNA is completed. While the mechanics of replication initiation are quite well understood, exactly what happens during termination is far from clear, although studies in bacterial and eukaryotic models over recent years have started to provide some insight. Bacterial models with a circular chromosome and a single bidirectional origin offer the distinct advantage that there is normally just one fusion event between two replication fork complexes as synthesis terminates. Moreover, whereas termination of replication appears to happen in many bacteria wherever forks happen to meet, termination in some bacterial species, including the well-studied bacteria Escherichia coli and Bacillus subtilis, is more restrictive and confined to a 'replication fork trap' region, making termination even more tractable. This region is defined by multiple genomic terminator (ter) sites, which, if bound by specific terminator proteins, form unidirectional fork barriers. In this review we discuss a range of experimental results highlighting how the fork fusion process can trigger significant pathologies that interfere with the successful conclusion of DNA replication, how these pathologies might be resolved in bacteria without a fork trap system and how the acquisition of a fork trap might have provided an alternative and cleaner solution, thus explaining why in bacterial species that have acquired a fork trap system, this system is remarkably well maintained. Finally, we consider how eukaryotic cells can cope with a much-increased number of termination events.

High bioavailability, short half-life, and metabolism into hydromorphone-3-glucuronide following single intramuscular and intravenous administration of hydromorphone hydrochloride to great horned owls (Bubo virginianus).

OBJECTIVE: To determine the pharmacokinetic parameters of hydromorphone hydrochloride and its metabolite, hydromorphone-3-glucuronide (H3G), after a single IV and IM dose in great horned owls (Bubo virginianus). ANIMALS: 6 healthy adult great horned owls (3 females and 3 males). PROCEDURES: A single dose of hydromorphone (0.6 mg/kg) was administered once IM (pectoral muscles) and IV (left jugular) with a 6-week washout period between experiments. Blood samples were collected at 5 minutes and 0.5, 1.5, 2, 3, 6, 9, and 12 hours after drug administration. Plasma hydromorphone and H3G concentrations were determined with liquid chromatography-tandem mass spectrometry, and a noncompartmental analysis was used for the determination of pharmacokinetic parameters. RESULTS: Hydromorphone had a high bioavailability of 170.8 ± 37.6% and rapid elimination after IM administration and rapid plasma clearance and a large volume of distribution after IV administration. Mean Cmax was 225.46 ± 0.2 ng/mL at 13 minutes after IM injection. Mean volume of distribution and plasma drug clearance was 4.29 ± 0.5 L/kg and 62.11 ± 14.6 mL/min/kg, respectively, after IV administration. Mean t1/2 was 1.62 ± 0.36 and 1.35 ± 0.59 hours after IM and IV administration, respectively. The metabolite H3G was readily measured shortly after administration by both routes. CLINICAL RELEVANCE: A single dose of 0.6 mg/kg was well tolerated in all birds. Hydromorphone rapidly attained plasma concentrations following IM administration and had high bioavailability and short t1/2. This study is the first to document the presence of the metabolite H3G in avian species, which suggests similar hydromorphone metabolism as in mammals.

Treatment of Pain in Birds.

This article provides an overview of the current understanding of evidence-based clinical analgesic use in birds. The field of avian analgesia has dramatically expanded during the last 20 years, affording more options for alleviating both acute and chronic pain. These options include opioids, nonsteroidal anti-inflammatory drugs, local anesthetics, and/or other drugs like gabapentin, amantadine, and cannabinoids, acting at different points in the nociceptive system thereby helping to provide greater pain relief while reducing the risk of adverse effects when combined.

Evaluation of Oral Transmucosal Administration of Pentobarbital for Euthanasia of Conscious Wild Birds.

This prospective study evaluated oral transmucosal pentobarbital sodium at three doses in 110 wild-caught wild birds requiring euthanasia. Birds received transmucosal pentobarbital at five (430 mg/kg), six (516 mg/kg), and seven times (602 mg/kg) the intravenous dose for mammals. Time to first effects and loss of consciousness, presence of pupillary light and corneal reflexes, apnea, and asystole were recorded each minute. When asystole was not achieved at 5 minutes, IV pentobarbital was administered. Combining data for all doses, loss of consciousness occurred at a median (range) of 2 minutes (0-4.75 min), apnea at 3 minutes (0-6 min), and asystole at 4 minutes (0.5-5 min). Loss of consciousness and apnea occurred significantly faster in the 602 mg/kg dose group than in the 430 mg/kg group (p = 0.009, difference of 0.6 ± 0.2 min; p = 0.024, difference of 0.7 ± 0.3 min), respectively. Apnea and asystole were achieved in 80/110 birds within 5 minutes. Oral transmucosal pentobarbital results in rapid loss of consciousness and respiratory arrest and provides a reliable alternative euthanasia method compared to intravenous administration.

Cannabis dosing and administration for sleep: a systematic review.

STUDY OBJECTIVES: As cannabis is increasingly used to treat sleep disorders, we performed a systematic review to examine the effects of cannabis on sleep and to guide cannabis prescribers in their recommendations to patients, specifically focusing on dosing. METHODS: We searched EMBASE, Medline, and Web of Science and identified 4550 studies for screening. Five hundred sixty-eight studies were selected for full-text review and 31 were included for analysis. Study results were considered positive based on improvements in sleep architecture or subjective sleep quality. Bias in randomized controlled trials was assessed using Cochrane Risk of Bias tool 2.0. RESULTS: Sleep improvements were seen in 7 out of 19 randomized studies and in 7 out of 12 uncontrolled trials. There were no significant differences between the effects of tetrahydrocannabinol and cannabidiol. Cannabis showed most promise at improving sleep in patients with pain-related disorders, as compared to those with neurologic, psychiatric, or sleep disorders, and showed no significant effects on healthy participants' sleep. While subjective improvements in sleep quality were often observed, diagnostic testing showed no improvements in sleep architecture. Adverse events included headaches, sedation, and dizziness, and occurred more frequently at higher doses, though no serious adverse events were observed. CONCLUSION: High-quality evidence to support cannabis use for sleep remains limited. Heterogeneity in cannabis types, doses, timing of administration, and sleep outcome measures limit the ability to make specific dosing recommendations.

Clinicopathological findings in and prognostic factors for domestic rabbits with liver lobe torsion: 82 cases (2010-2020).

OBJECTIVE: To document clinicopathologic findings in domestic rabbits with liver lobe torsion and identify prognostic factors. ANIMALS: 82 rabbits. PROCEDURE: Medical records of 4 institutions were reviewed to identify rabbits with an antemortem diagnosis of liver lobe torsion that were examined between 2010 and 2020. RESULTS: The prevalence of liver lobe torsion was 0.7% (82/11,402). In all 82 rabbits, the diagnosis was made by means of abdominal ultrasonography. Fifty (60.1%) rabbits underwent liver lobectomy, 23 (28%) received medical treatment alone, and 9 (10.9%) were euthanized or died on presentation. Overall, 32 (39%) rabbits died within 7 days of initial presentation and 50 (61%) survived. Seven-day survival rate did not differ significantly between medical treatment alone and surgical treatment. However, median survival time following medical treatment (530 days) was shorter than that following surgical treatment (1,452 days). Six of 14 rabbits had evidence of systemic inflammatory disease on necropsy. Rabbits with right liver lobe torsion were less likely to survive for 7 days than were those with caudate torsions (P = 0.046; OR, 3.27; 95% CI, 1.04 to 11.3). Rabbits with moderate to severe anemia were less likely to survive for 7 days than were rabbits that were not anemic or had mild anemia (P = 0.006; OR, 4.41; 95% CI, 1.55 to 12.51). Other factors associated with a decreased 7-day survival rate were high heart rate at admission (P = 0.013) and additional days without defecation after admission (P < 0.001). Use of tramadol was associated with an increased survival rate (P = 0.018). CLINICAL RELEVANCE: The prognosis for rabbits with liver lobe torsions was more guarded than previously described. Rabbits that underwent liver lobectomy had a longer median survival time than did rabbits that only received medical treatment.

Escherichia coli Chromosome Copy Number Measurement Using Flow Cytometry Analysis.

Flow cytometry is a high-throughput technique that analyzes individual particles as they pass through a laser beam. These particles can be individual cells and by detecting cell-scattered light their number and relative size can be measured as they pass through the beam. Labeling of molecules, usually via a fluorescent reporter, allows the amount of these molecules per cell to be quantified. DNA content can be estimated using this approach and here we describe how flow cytometry can be used to assess the DNA content of Escherichia coli cells.

Absorption of grapiprant in red-tailed hawks (Buteo jamaicensis) is decreased when administered with food.

OBJECTIVE: Describe the pharmacokinetics of grapiprant administered orally with food to red-tailed hawks (RTHAs; Buteo jamaicensis) and compare the results with previously described grapiprant pharmacokinetics administered without food in this species. ANIMALS: 6 healthy adult RTHA (3 males, 3 females) under human care. PROCEDURES: A single dose of grapiprant (30 mg/kg) was given orally to RTHAs, followed by force-feeding. Blood samples were obtained at 14 time points for 120 hours postgrapiprant administration. Plasma concentrations of grapiprant were measured via tandem liquid chromatography-mass spectrometry. Nonparametric superimposition using pharmacokinetic modeling software used plasma concentrations to calculate simulations of grapiprant plasma concentrations for 30 mg/kg administered orally with food every 12 hours. RESULTS: The arithmetic mean maximum plasma concentration was 405.8 ng/mL, time to maximum plasma concentration was 16 hours, and harmonic mean terminal half-life was 15.6 hours. Simulations determined 30 mg/kg every 12 hours could attain minimum effective concentrations (> 164 ng/mL) reported in dogs for a sustained period of approximately 20 hours. CLINICAL RELEVANCE: Grapiprant plasma concentrations were achieved above the canine therapeutic concentrations within 16 hours postmedication. Mean concentrations were maintained for approximately 20 hours. Simulations support a dosing frequency of 12-hour intervals with food reaching minimum effective concentrations established for canines, although it is unknown whether these plasma concentrations are therapeutic for birds. Bioaccumulation was not noted on simulations secondary to increased grapiprant administration. Further research including multidose assessments at this current dose with food, in vitro pharmacological characterization, and pharmacodynamic studies in this species are warranted.

Pharmacokinetics of pimobendan following oral administration to New Zealand White rabbits (Oryctolagus cuniculus).

OBJECTIVE: To determine the pharmacokinetics and potential adverse effects of pimobendan after oral administration in New Zealand White rabbits (Ocytolagus cuniculi). ANIMALS: 10 adult sexually intact (5 males and 5 females) rabbits. PROCEDURES: 2 pilot studies were performed with a pimobendan suspension or oral tablets. Eight rabbits received 7.5 mg of pimobendan (mean 2.08 mg/kg) suspended in a critical care feeding formula. Plasma concentrations of pimobendan and O-demethylpimobendan (ODMP) were measured, and pharmacokinetic parameters were calculated for pimobendan by noncompartmental analysis. Body weight, food and water consumption, mentation, urine, and fecal output were monitored. RESULTS: Mean ± SD maximum concentration following pimobendan administration was 15.7 ± 7.54 ng/mL and was detected at 2.79 ± 1.25 hours. The half-life was 3.54 ± 1.32 hours. Plasma concentrations of pimobendan were detectable for up to 24 hours. The active metabolite, ODMP, was detected in rabbits for 24 to 36 hours. An adverse event occurred following administration of pimobendan in tablet form in 1 pilot study, resulting in death secondary to aspiration. No other adverse events occurred. CLINICAL RELEVANCE: Plasma concentrations of pimobendan were lower than previously reported for dogs and cats, despite administration of higher doses, and had longer time to maximum concentration and half-life. Based on this study, 2 mg/kg of pimobendan in a critical care feeding formulation should maintain above a target plasma concentration for 12 to 24 hours. However, further studies evaluating multiple-dose administration as well as pharmacodynamic studies and clinical trials in rabbits with congestive heart failure are needed to determine accurate dose and frequency recommendations.

Raptor Sedation and Anesthesia.

Sedation and/or anesthesia is routinely and successfully used in raptors for a wide variety of procedures from the routine such as physical examination, radiographs, or venipuncture, to the more complex, such as orthopedic surgeries. Understanding the anatomy and physiology of raptor patients who present for care, and being fully prepared before the start of any procedure, can increase the success of anesthetic procedures. Recent advances in raptor sedation and anesthesia continue to improve the health and welfare of these avian patients.

Diagnosis and treatment of Candida glabrata proventriculitis in an eclectus parrot (Eclectus roratus).

CASE DESCRIPTION: An 8-year-old sexually intact female eclectus parrot (Eclectus roratus) with a 4-day history of hyporexia and lethargy and a 1-day history of tenesmus was examined. CLINICAL FINDINGS: Severe leukocytosis characterized by severe heterophilia and moderate monocytosis was present. Marked dilation of the proventriculus and ventriculus and ascites were identified by means of radiography, coelomic ultrasonography, and contrast-enhanced CT, with no clinically relevant motility noted on ultrasonography. Results of coelomic fluid analysis were consistent with pyogranulomatous effusion. Endoscopy of the upper gastrointestinal tract following proventricular and ventricular lavage showed a thick caseous plaque occupying 30% of the caudal proventricular mucosa. Abundant yeast organisms were evident during cytologic examination of a proventricular and ventricular wash sample, and fecal culture yielded Candida glabrata. TREATMENT AND OUTCOME: The bird was treated with SC fluids, assisted feedings, nystatin, fluconazole, amoxicillin-clavulanic acid, enrofloxacin, gastroprotectants, maropitant, and analgesics and slowly improved during hospitalization. A marked decrease in proventricular dilation was evident on serial radiographs obtained over a 12-month period. One year after diagnosis, the bird was presented with a 1-week history of hyporexia and lethargy, and fecal culture grew C glabrata. Antifungal treatment was resumed for 3 months. The bird had no clinical signs of infection 16 months after this recurrence, and subsequent fecal cultures were negative for fungal growth. CLINICAL RELEVANCE: Findings illustrate the importance of upper gastrointestinal endoscopy in diagnosing proventricular and ventricular dilation in birds and emphasize the need for long-term antifungal treatment and monitoring in birds with fungal infections.

Termination of DNA replication at Tus-ter barriers results in under-replication of template DNA.

The complete and accurate duplication of genomic information is vital to maintain genome stability in all domains of life. In Escherichia coli, replication termination, the final stage of the duplication process, is confined to the "replication fork trap" region by multiple unidirectional fork barriers formed by the binding of Tus protein to genomic ter sites. Termination typically occurs away from Tus-ter complexes, but they become part of the fork fusion process when a delay to one replisome allows the second replisome to travel more than halfway around the chromosome. In this instance, replisome progression is blocked at the nonpermissive interface of the Tus-ter complex, termination then occurs when a converging replisome meets the permissive interface. To investigate the consequences of replication fork fusion at Tus-ter complexes, we established a plasmid-based replication system where we could mimic the termination process at Tus-ter complexes in vitro. We developed a termination mapping assay to measure leading strand replication fork progression and demonstrate that the DNA template is under-replicated by 15 to 24 bases when replication forks fuse at Tus-ter complexes. This gap could not be closed by the addition of lagging strand processing enzymes or by the inclusion of several helicases that promote DNA replication. Our results indicate that accurate fork fusion at Tus-ter barriers requires further enzymatic processing, highlighting large gaps that still exist in our understanding of the final stages of chromosome duplication and the evolutionary advantage of having a replication fork trap.

Pharmacokinetics of grapiprant administered to red-tailed hawks (Buteo jamaicensis) after food was withheld for 24 hours.

OBJECTIVE: To identify an oral dose of grapiprant for red-tailed hawks (RTHAs; Buteo jamaicensis) that would achieve a plasma concentration > 164 ng/mL, which is considered therapeutic for dogs with osteoarthritis. ANIMALS: 6 healthy adult RTHAs. PROCEDURES: A preliminary study, in which grapiprant (4 mg/kg [n = 2], 11 mg/kg [2], or 45 mg/kg [2]) was delivered into the crop of RTHAs from which food had been withheld for 24 hours, was performed to obtained pharmacokinetic data for use with modeling software to simulate results for grapiprant doses of 20, 25, 30, 35, and 40 mg/kg. Simulation results directed our selection of the grapiprant dose administered to the RTHAs in a single-dose study. Plasma grapiprant concentration, body weight, and gastrointestinal signs of RTHAs were monitored. RESULTS: On the basis of results from the preliminary study and simulations, a grapiprant dose of 30 mg/kg was used in the single-dose study. The geometric mean maximum observed plasma concentration of grapiprant was 3,184 ng/mL, time to maximum plasma grapiprant concentration was 2.0 hours, and the harmonic mean terminal half-life was 17.1 hours. No substantial adverse effects were observed. CONCLUSIONS AND CLINICAL RELEVANCE: Although the single dose of grapiprant (30 mg/kg) delivered into the crop achieved plasma concentrations > 164 ng/mL in the RTHAs, it was unknown whether this concentration would be therapeutic for birds. Further research that incorporates multidose assessments, safety monitoring, and pharmacodynamic data collection is warranted on the use of grapiprant in RTHAs from which food was withheld versus not withheld.

Evaluation of Atherosclerotic Lesions and Risk Factors of Atherosclerosis in Raptors in Northern California.

Atherosclerosis is a chronic inflammatory disease that has been reported to affect the cardiovascular system of many avian species. However, atherosclerosis in raptor species has not been fully evaluated. The aim of this study was to histologically characterize central and peripheral atherosclerotic lesions in raptors that were submitted to a pathology service in Northern California from 1986 to 2013. We also evaluated risk factors, including age, gender, origin, and avian family. Atherosclerotic lesions were categorized as minimal, mild, moderate, or severe, based on the severity of the lesions and their distribution within the arterial wall. Among the central arteries, lesions were determined to be of greater severity in the aorta than in the pulmonary artery. More than 50% of the peripheral arteries were affected, including 53.1% (17/32) myocardial, 52% (13/25) coronary, 62.9% (22/35) arteries in the kidney, 52.2% (12/23) gonadal and 51.7% (15/29) splenic arteries; however, hepatic and pulmonary arteries were uncommonly affected. Atherosclerosis was diagnosed in 17 raptor species representing 4 families: Accipitridae, Cathartidae, Falconidae, and Strigidae. The overall prevalence (95% CI) of atherosclerosis in raptors was 2.3% (36/1574; range, 1.63%-3.19%) with the Falconidae having the highest prevalence at 7.4% (9/122; range 3.64%-13.93%) and with 0% detected in the Tytonidae and Pandionidae families. A multiple logistic regression model that jointly accounted for differences in risk by family, age, and gender found that the risk in Accipitridae was significantly less than that of Falconidae, that adult raptors were at greater risk of atherosclerotic lesions than juveniles were, and that females were more frequently affected than males were.

Chlamydia buteonis in birds of prey presented to California wildlife rehabilitation facilities.

Chlamydial infections, caused by a group of obligate, intracellular, gram-negative bacteria, have health implications for animals and humans. Due to their highly infectious nature and zoonotic potential, staff at wildlife rehabilitation centers should be educated on the clinical manifestations, prevalence, and risk factors associated with Chlamydia spp. infections in raptors. The objectives of this study were to document the prevalence of chlamydial DNA shedding and anti-chlamydial antibodies in raptors admitted to five wildlife rehabilitation centers in California over a one-year period. Chlamydial prevalence was estimated in raptors for each center and potential risk factors associated with infection were evaluated, including location, species, season, and age class. Plasma samples and conjunctiva/choana/cloaca swabs were collected for serology and qPCR from a subset of 263 birds of prey, representing 18 species. Serologic assays identified both anti-C. buteonis IgM and anti-chlamydial IgY antibodies. Chlamydial DNA and anti-chlamydial antibodies were detected in 4.18% (11/263) and 3.14% (6/191) of patients, respectively. Chamydial DNA was identified in raptors from the families Accipitridae and Strigidae while anti-C.buteonis IgM was identified in birds identified in Accipitridae, Falconidae, Strigidae, and Cathartidae. Two of the chlamydial DNA positive birds (one Swainson's hawk (Buteo swainsoni) and one red-tailed hawk (Buteo jamaicensis)) were necropsied, and tissues were collected for culture. Sequencing of the cultured elementary bodies revealed a chlamydial DNA sequence with 99.97% average nucleotide identity to the recently described Chlamydia buteonis. Spatial clusters of seropositive raptors and raptors positive for chlamydial DNA were detected in northern California. Infections were most prevalent during the winter season. Furthermore, while the proportion of raptors testing positive for chlamydial DNA was similar across age classes, seroprevalence was highest in adults. This study questions the current knowledge on C. buteonis host range and highlights the importance of further studies to evaluate the diversity and epidemiology of Chlamydia spp. infecting raptor populations.

A Fork Trap in the Chromosomal Termination Area Is Highly Conserved across All Escherichia coli Phylogenetic Groups.

Termination of DNA replication, the final stage of genome duplication, is surprisingly complex, and failures to bring DNA synthesis to an accurate conclusion can impact genome stability and cell viability. In Escherichia coli, termination takes place in a specialised termination area opposite the origin. A 'replication fork trap' is formed by unidirectional fork barriers via the binding of Tus protein to genomic ter sites. Such a fork trap system is found in some bacterial species, but it appears not to be a general feature of bacterial chromosomes. The biochemical properties of fork trap systems have been extensively characterised, but little is known about their precise physiological roles. In this study, we compare locations and distributions of ter terminator sites in E. coli genomes across all phylogenetic groups, including Shigella. Our analysis shows that all ter sites are highly conserved in E. coli, with slightly more variability in the Shigella genomes. Our sequence analysis of ter sites and Tus proteins shows that the fork trap is likely to be active in all strains investigated. In addition, our analysis shows that the dif chromosome dimer resolution site is consistently located between the innermost ter sites, even if rearrangements have changed the location of the innermost termination area. Our data further support the idea that the replication fork trap has an important physiological role that provides an evolutionary advantage.

Effectiveness of glass beads for plating cell cultures.

Cell plating, the spreading out of a liquid suspension of cells on a surface followed by colony growth, is a common laboratory procedure in microbiology. Despite this, the exact impact of its parameters on colony growth has not been extensively studied. A common protocol involves the shaking of glass beads within a Petri dish containing solid growth media. We investigated the effects of multiple parameters in this protocol: the number of beads, the shape of movement, and the number of movements. Standard suspensions of Escherichia coli were spread while varying these parameters to assess their impact on colony growth. Results were assessed by a variety of metrics: the number of colonies, the mean distance between closest colonies, and the variability and uniformity of their spatial distribution. Finally, we devised a mathematical model of shifting billiard to explain the heterogeneities in the observed spatial patterns. Exploring the parameters that affect the most fundamental techniques in microbiology allows us to better understand their function, giving us the ability to precisely control their outputs for our exact needs.

Release Rates and Complications for Birds of Prey With Antebrachial Fractures at a Veterinary Teaching Hospital.

A retrospective case series that included 253 free-ranging birds of prey admitted to a rehabilitation center was conducted to describe the treatment and outcome of antebrachial fractures. Medical records from birds of prey belonging to 21 species admitted with antebrachial fracture between 1989 and 2015 at the University of California, Davis, were reviewed. Species distribution on admission, treatment, outcome, and complications were described by fracture category and species. Among 134 birds treated after initial triage on the day of admission, 4 bone/ wing categories were identified: 83 birds had an ulnar fracture only; 18 birds had a radial fracture only; 28 birds had a concomitant fracture of the radius and ulna on the same wing; and 5 birds had bilateral antebrachial fractures. Logistic regressions were performed to determine which factors were associated with a positive outcome within each of these 4 categories. Among birds having only an ulnar fracture, those with a closed fracture were significantly more likely to be released than birds with open fractures (P = .03; odds ratio = 5.43, 95% confidence interval: 1.29-28.12). In addition, birds with a fracture of the middle third of the ulna were significantly more likely to be released than birds diagnosed with a single fracture of the proximal third of the ulna (P = .02; odds ratio = 4.54, 95% confidence interval: 1.35-16.64). No significant prognostic factor was detected in other fracture categories.