Adaptive strategies of Yersinia pestis to persist during inter-epizootic and epizootic periods

Plague is a flea-borne zoonotic bacterial disease caused by Yersinia pestis. It has caused three historical pandemics, including the Black Death which killed nearly a third of Europe's population in the 14th century. In modern times, plague epizootics can extirpate entire susceptible wildlife populations and then disappear for long time periods. Understanding how Y. pestis is maintained during inter-epizootic periods and the factors responsible for transitioning to epizootics is important for preventing and controlling pathogen transmission and ultimately reducing the burden of human disease. In this review, we focus primarily on plague in North American foci and discuss the potential adaptive strategies Y. pestis might employ to ensure not only its survival during inter-epizootic periods but also the rapid epizootic spread and invasion of new territories that are so characteristic of plague and have resulted in major pandemics and establishment of plague foci throughout much of the world.     

Apparent adiposity assessed by standardised scoring systems and morphometric measurements in horses and ponies

This study described a scoring system for the assessment of apparent neck adiposity and evaluated morphometric measurements for assessment of neck and overall adiposity. Twenty-one barren Thoroughbred mares, 13 Arabian geldings and 75 Welsh, Dartmoor, or crossbred pony mares, were clinically examined and blood samples analysed for insulin, glucose, leptin, and triglycerides. Bodyweight (BW), height, length, girth and abdominal circumferences, neck length, neck crest height and neck circumference were measured, and body condition scores (BCS) and cresty neck scores (CNS) were rated. Girth:height ratio had the strongest associations with BCS (r(s)=0.64, P<0.001 in horses; r(s)=0.83, P<0.001 in ponies) and blood variables, such as leptin (r(s)=0.39, P=0.024 in horses; r(s)=0.68, P<0.001 in ponies). Crest height and neck circumference:height ratio had the strongest association with CNS (r(s)>0.50, P<0.01) and blood variables, such as insulin (r(s)0.40, P<0.05). Cresty neck score was useful in the assessment of neck crest adiposity and had physiological relevance, as demonstrated by associations with blood variables. Girth:height was the most suitable morphometric for assessment of overall adiposity, and either crest height or neck circumference:height was a suitable morphometric for assessment of apparent neck adiposity.     

Shortcuts for marine conservation planning: The effectiveness of socioeconomic data surrogates

Marine protected area (MPA) networks designed without consideration of the interests of local communities are likely to fail. However, in many regions where conservation action is needed most urgently, socioeconomic data are not available at spatial scales relevant to conservation planning. In the Philippines, the primary stakeholders relevant to conservation efforts in coastal waters are small-scale fishers. Unlike commercial fisheries, no logbook data are kept to record fishers’ spatial effort and usage patterns. We investigated the effects of including different surrogates for small-scale fishing effort in the systematic design of an MPA network for Siquijor Province. We compared a reserve selection scenario in which socioeconomic data were not considered with four different surrogates for fishing effort and with empirical data on the spatial distribution of fishing effort collected through interviews. We assumed that minimising opportunity costs to fishers would increase the likelihood that they would support and comply with MPA implementation, resulting in more effective conservation. Surrogates modelled on the number of fishers or boats in each community consistently outperformed those based on population census data. However, none of the surrogates we tested were able to accurately predict fine-scale resource use patterns. Whilst socioeconomic surrogates may be able to assist conservation planners to identify regional-scale opportunities where conservation objectives may be met more easily, they cannot act as a shortcut for comprehensive consultation with communities, which will be required to identify actual sites for MPA implementation.     

Characterization of protein changes associated with sugar beet (Beta vulgaris) resistance and susceptibility to Fusarium oxysporum

Fusarium oxysporum (F-19) is a serious threat to sugar beet. Resistance exists, but the basis for resistance and disease is unknown. Protein extracts from sugar beet genotypes C1200.XH024 (resistant, R) and Fus7 (susceptible, S) were analyzed by multidimensional liquid chromatography at 2 and 5 days postinoculation (dpi) and compared to mock-inoculated controls. One hundred twenty-one (R) and 73 (S) protein peaks were induced/repressed by F-19, approximately 12 (R) and 8% (S) of the total proteome detected. Temporal protein regulation occurred within and between each genotype, indicating that the timing of expression may be important for resistance. Thirty-one (R) and 48 (S) of the differentially expressed peaks were identified using matrix-assisted laser desorption-ionization with tandem time-of-flight mass spectrometry; others were below detection level. Comparison between the two genotypes uncovered R- and S-specific proteins with potential roles in resistance and disease development, respectively. Use of these proteins to select for new sources of resistance and to develop novel disease control strategies is discussed.     

Effects of different cyclooxygenase inhibitors on prostaglandin E2 production,steroidogenesis and ovulation of bovine preovulatory follicles

Ovulation is an inflammation-like process, and cyclooxygenase-2 (COX-2)-dependent production of prostaglandin E₂ (PGE₂) is its key mediator. Balanced regulation of inflammatory processes in high-yielding dairy cows may be essential for physiological ovulation and fertility. This study aimed to elucidate the mechanisms underlying ovulation failure and cyst development after disturbing intrafollicular inflammatory cascades. Therefore, nonselective (indomethacin and flunixin-meglumine), COX-2 selective (meloxicam), and highly COX-2 selective (NS-398) inhibitors were injected into preovulatory follicles 16 h after administration of GnRH, and ovulation was monitored via ultrasound examination. Additionally, follicular fluid was collected after injection of indomethacin, meloxicam, and NS-398. Moreover, primary granulosa cell cultures from preovulatory follicles were prepared and treated with indomethacin, meloxicam, and NS-398. The concentrations of 17β-estradiol, progesterone, and prostaglandin E₂ (PGE₂) in the follicular fluid and cell supernatant were estimated. Indomethacin and flunixin-meglumine blocked ovulation, even at low doses, and led to ovarian cyst development. The selective and highly selective COX-2 inhibitors meloxicam and NS-398 were not effective in blocking ovulation. However, indomethacin, meloxicam, and NS-398 significantly and comparably reduced PGE₂ concentration in vivo and in vitro (P < 0.05) but had no effect on estradiol or progesterone production. This may contradict the generally accepted hypothesis that PGE₂ is a key mediator of ovulation and progesterone production. Our results suggest a connection between ovarian disorders and inflammatory actions in early postpartum cows.     

Ionized hypercalcemia in cats with azotemic chronic kidney disease (2012‐2018)

BackgroundHypercalcemia is associated with chronic kidney disease (CKD) in cats, but studies assessing the physiologically relevant ionized calcium fraction are lacking.ObjectivesTo describe the prevalence and incidence rate of ionized hypercalcemia, and to explore predictor variables to identify cats at risk of ionized hypercalcemia in a cohort of cats diagnosed with azotemic CKD.AnimalsOne hundred sixty‐four client‐owned cats with azotemic CKD.MethodsVariables independently associated with ionized hypercalcemia at diagnosis of azotemic CKD were explored by binary logistic regression. Cats that were normocalcemic at diagnosis of azotemic CKD were followed over a 12‐month period or until ionized hypercalcemia occurred and baseline predictor variables for ionized hypercalcemia explored using Cox proportional hazards and receiver operating characteristic curve analysis.ResultsIonized hypercalcemia (median, 1.41 mmol/L; range, 1.38‐1.68) was observed in 33/164 (20%) cats at diagnosis of azotemic CKD and was associated with male sex, higher plasma total calcium and potassium concentrations, and lower plasma parathyroid hormone concentrations. Twenty‐five of 96 initially normocalcemic (26%) cats followed for minimum 90 days developed ionized hypercalcemia (median, 1.46 mmol/L; range, 1.38‐1.80) at a median of 140 days after diagnosis of azotemic CKD (incidence rate, 0.48 per feline patient‐year). Only body condition score was independently associated with incident ionized hypercalcemia.Conclusions and Clinical ImportanceThe occurrence of ionized hypercalcemia is high in cats with CKD. Continued monitoring of blood ionized calcium concentrations is advised.     

Prevalence and clinical features of thoracolumbar intervertebral disc‐associated epidural hemorrhage in dogs

BackgroundIntervertebral disc‐associated epidural hemorrhage (EH) in dogs is a poorly understood neurological condition.ObjectiveTo compare the clinical presentation, magnetic resonance imaging (MRI) changes, and clinical outcome of dogs with acute thoracolumbar intervertebral disc herniation (TL‐IVDH) with and without EH.AnimalsOne hundred sixty client‐owned dogs that underwent MRI and hemilaminectomy for acute TL‐IVDH at a private practice in Colorado, including 63 dogs with EH and 97 dogs without EH.MethodsRetrospective review of medical record data from 160 dogs presenting sequentially to a single practice with acute TL‐IVDH that underwent MRI and hemilaminectomy surgery.ResultsSixty‐three of 160 (39%) dogs had confirmed EH. French Bulldogs were significantly overrepresented (23/63; odds ratio [OR]: 4.1; 95% confidence interval [CI]: 1.8‐9.0; P < .001) of the EH cases. Dogs with EH were more likely to present with clinical signs less than 48 hours than were dogs without EH (24‐48 vs 48‐72 hours; OR: 2.4; 95% CI: 1.2‐4.6; P = .02) and were more likely to be nonambulatory on presentation (OR: 2.1; 95% CI: 1.0‐4.1; P = .04). Dogs with EH were more likely to have <50% cross‐sectional spinal cord compression than dogs without EH (OR: 2.3 vs. 0.4; 95% CI: 1.2‐4.4 and 0.2‐0.9, respectively), longer longitudinal spinal cord compression (3 spaces vs 1 space, P < .001), and greater intrinsic spinal cord change (grade 3/severe vs grade 1/mild; P < .001) based on MRI. The location of the intervertebral disc herniation in French Bulldogs with EH was more likely to be thoracolumbar (OR: 10.8; 95% CI: 2.1‐55.7; P = .03).Conclusions and Clinical ImportanceFrench Bulldogs have a high prevalence of intervertebral disc‐associated EH. Dogs with EH have a shorter clinical course and are more likely to be nonambulatory on initial presentation.     

Evolution of a microbial acetyltransferase for modification of glyphosate: a novel tolerance strategy

N-Acetylation is a modification of glyphosate that could potentially be used in transgenic crops, given a suitable acetyltransferase. Weak enzymatic activity (k(cat) = 5 min(-1), K(M) = 1 mM) for N-acetylation of glyphosate was discovered in several strains of Bacillus licheniformis (Weigmann) Chester by screening a microbial collection with a mass spectrometric assay. The parental enzyme conferred no tolerance to glyphosate in any host when expressed as a transgene. Eleven iterations of DNA shuffling resulted in a 7000-fold improvement in catalytic efficiency (k(cat)/K(M)), sufficient for conferring robust tolerance to field rates of glyphosate in transgenic tobacco and maize. In terms of k(cat)/K(M), the native enzyme exhibited weak activity (4-450% of that with glyphosate) with seven of the common amino acids. Evolution of the enzyme towards an improved k(cat)/K(M) for glyphosate resulted in increased activity toward aspartate (40-fold improved k(cat)), but activity with serine and phosphoserine almost completely vanished. No activity was observed among a broad sampling of nucleotides and antibiotics. Improved catalysis with glyphosate coincided with increased thermal stability.     

Antioxidant 1 copper chaperone gene expression and copper levels in dog osteosarcoma patients

Twenty-four dogs with OS underwent limb amputation. Serum, OS tumour, and normal bone were harvested at time of surgery. RNA was extracted and gene expression was performed using quantitative polymerase chain reaction (qPCR). Tissue and blood copper concentrations were also determined with spectrophotometry. Compared to bone, tumour samples had significantly higher expressions of antioxidant 1 copper chaperone (ATOX1, p = .0003). OS tumour copper levels were significantly higher than that of serum (p < .010) and bone (p = .038). Similar to our previous observations in mouse and human OS, dog OS demonstrates overexpression of genes that regulate copper metabolism (ATOX1), and subsequent copper levels. Dogs with OS may provide a robust comparative oncology platform for the further study of these factors, as well as potential pharmacologic interventions.