Limited effect of thermal pruning on wild blueberry crop and its root-associated microbiota.

Thermal pruning was a common pruning method in the past but has progressively been replaced by mechanical pruning for economic reasons. Both practices are known to enhance and maintain high yields; however, thermal pruning was documented to have an additional sanitation effect by reducing weeds and fungal diseases outbreaks. Nevertheless, there is no clear consensus on the optimal fire intensity required to observe these outcomes. Furthermore, fire is known to alter the soil microbiome as it impacts the soil organic layer and chemistry. Thus far, no study has investigated into the effect of thermal pruning intensity on the wild blueberry microbiome in agricultural settings. This project aimed to document the effects of four gradual thermal pruning intensities on the wild blueberry performance, weeds, diseases, as well as the rhizosphere fungal and bacterial communities. A field trial was conducted using a block design where agronomic variables were documented throughout the 2-year growing period. MiSeq amplicon sequencing was used to determine the diversity as well as the structure of the bacterial and fungal communities. Overall, yield, fruit ripeness, and several other agronomical variables were not significantly impacted by the burning treatments. Soil phosphorus was the only parameter with a significant albeit temporary change (1 month after thermal pruning) for soil chemistry. Our results also showed that bacterial and fungal communities did not significantly change between burning treatments. The fungal community was dominated by ericoid mycorrhizal fungi, while the bacterial community was mainly composed of Acidobacteriales, Isosphaerales, Frankiales, and Rhizobiales. However, burning at high intensities temporarily reduced Septoria leaf spot disease in the season following thermal pruning. According to our study, thermal pruning has a limited short-term influence on the wild blueberry ecosystem but may have a potential impact on pests (notably Septoria infection), which should be explored in future studies to determine the burning frequency necessary to control this disease.

Conditioning Machine Learning Models to Adjust Lowbush Blueberry Crop Management to the Local Agroecosystem.

Agroecosystem conditions limit the productivity of lowbush blueberry. Our objectives were to investigate the effects on berry yield of agroecosystem and crop management variables, then to develop a recommendation system to adjust nutrient and soil management of lowbush blueberry to given local meteorological conditions. We collected 1504 observations from N-P-K fertilizer trials conducted in Quebec, Canada. The data set, that comprised soil, tissue, and meteorological data, was processed by Bayesian mixed models, machine learning, compositional data analysis, and Markov chains. Our investigative statistical models showed that meteorological indices had the greatest impact on yield. High mean temperature at flower bud opening and after fruit maturation, and total precipitation at flowering stage showed positive effects. Low mean temperature and low total precipitation before bud opening, at flowering, and by fruit maturity, as well as number of freezing days (<-5 °C) before flower bud opening, showed negative effects. Soil and tissue tests, and N-P-K fertilization showed smaller effects. Gaussian processes predicted yields from historical weather data, soil test, fertilizer dosage, and tissue test with a root-mean-square-error of 1447 kg ha-1. An in-house Markov chain algorithm optimized yields modelled by Gaussian processes from tissue test, soil test, and fertilizer dosage as conditioned to specified historical meteorological features, potentially increasing yield by a median factor of 1.5. Machine learning, compositional data analysis, and Markov chains allowed customizing nutrient management of lowbush blueberry at local scale.

Evaluation of a Model of Long-Term Middle Ear Catheterization for Repeat Infusion Administration and Cochlear Hair Cell Injury in Guinea Pigs.

Middle ear administration has numerous applications, including antibiotherapy and gene therapy, and is increasingly used to target the auditory and vestibular systems. In animal studies, investigating repeated exposure that mimics clinical dosing regimens has remained a challenge due to the lack of suitable models. Intratympanic injections are not suitable for long-term studies due to the increased risk related to tympanic membrane rupture or scarring and repeat anesthesia events. Surgical models of middle ear catheterization previously used have not been reliable for longer than 4 weeks, resulted in elevated stress levels, and have been associated with significant changes related to the surgery and/or the presence of the catheter such as local trauma and inflammatory and degenerative processes. These complications cause decreased hearing/deafness and greatly diminish the value and accuracy of ototoxicity studies. We describe here a procedure that permits repeat dosing into the middle ear of guinea pigs and can be used to produce a model of aminoglycoside-induced hair cell injury. The innocuity of the procedures and the efficacy of the ototoxicity model were confirmed using auditory brain stem response assessment, histopathological evaluation, and cytocochleograms. Procedure-related changes were limited to minimal inflammation in the middle ear.

How plant allometry influences bud phenology and fruit yield in two Vaccinium species.

BACKGROUND AND AIMS: Understanding how plant allometry, plant architecture and phenology contribute to fruit production can identify those plant traits that maximize fruit yield. In this study, we compared these variables and fruit yield for two shrub species, Vaccinium angustifolium and Vaccinium myrtilloides, to test the hypothesis that phenology is linked to the plants' allometric traits, which are predictors of fruit production. METHODS: We measured leaf and flower phenology and the above-ground biomass of both Vaccinium species in a commercial wild lowbush blueberry field (Quebec, Canada) over a 2-year crop cycle; 1 year of pruning followed by 1 year of harvest. Leaf and flower phenology were measured, and the allometric traits of shoots and buds were monitored over the crop cycle. We hand-collected the fruits of each plant to determine fruit attributes and biomass. KEY RESULTS: During the harvesting year, the leafing and flowering of V. angustifolium occurred earlier than that of V. myrtilloides. This difference was related to the allometric characteristics of the buds due to differences in carbon partitioning by the plants during the pruning year. Through structural equation modelling, we identified that the earlier leafing in V. angustifolium was related to a lower leaf bud number, while earlier flowering was linked to a lower number of flowers per bud. Despite differences in reproductive allometric traits, vegetative biomass still determined reproductive biomass in a log-log scale model. CONCLUSIONS: Growing buds are competing sinks for non-structural carbohydrates. Their differences in both number and characteristics (e.g. number of flowers per bud) influence levels of fruit production and explain some of the phenological differences observed between the two Vaccinium species. For similar above-ground biomass, both Vaccinium species had similar reproductive outputs in terms of fruit biomass, despite differences in reproductive traits such as fruit size and number.

Preclinical testing of AAV9-PHP.B for transgene expression in the non-human primate cochlea.

In a number of mouse models of hereditary deafness, therapeutic transgene delivery to the cochlea and vestibular organs using adeno-associated viral vectors (AAVs) has shown striking rescue of hearing and balance. However, only a subset of AAV capsids have shown efficacy in transducing both inner hair cells and outer hair cells, and it is also not clear which of these can be translated to treatment of human inner ear. We recently reported efficient transgene expression of a GFP reporter in a non-human primate cochlea, in both inner and outer hair cells, following injection of the AAV9 capsid variant PHP.B via the round window membrane (RWM). However efficiency was poor at a lower dose. To further define the transduction potential of AAV9-PHP.B, we have performed a dosing study in the cynomolgus monkey and assessed vector-encoded GFP expression. Three animals were injected in both ears and four doses were tested. We describe a transmastoid surgical approach needed to access the RWM of this common primate model. We found that AAV9-PHP.B transduced nearly 100% of both IHCs and OHCs, from base to apex, at the higher doses (3.5 × 1011 and 7 × 1011 vector genomes). However, at lower doses there was a steep reduction in viral transduction. Thus, AAV9-PHP.B efficiently transduces the IHCs and OHCs of nonhuman primates, and should be considered as an AAV capsid for inner ear gene therapy in humans.

Lowbush blueberry fruit yield and growth response to inorganic and organic N-fertilization when competing with two common weed species.

Inorganic N fertilizers are commonly used in commercial blueberry fields; however, this form of N can favor increased weed species' growth, which can ultimately reduce the benefits of fertilization. We hypothesized that chipped ramial wood (CRW) compost is an effective alternative organic fertilizer for blueberry plants when weeds are present, as ericaceous shrub species are generally more efficient in utilizing organic N than herbaceous weed species. In this study, we measured the growth, fruit yield, and foliar N response of lowbush blueberry (Vaccinium angustifolium Aiton) to an application of 45 kg N ha-1 in the form of organic (CRW) or inorganic N (ammonium sulfate) in two areas of a commercial field colonized by either poverty oat grass (Danthonia spicata (L.) Beauv.) or sweet fern (Comptonia peregrina (L.) Coult.). We also assessed the impact of the fertilization treatments on litter decomposition rates. Contrary to our hypothesis, we found no significant increase in blueberry fruit yield or growth using CRW. By contrast, inorganic N-fertilization increased fruit yield by 70%. The effect was higher in the area colonized by D. spicata (+83%) than by C. peregrina (+45%). Blueberry fruit yield was on average twice higher in the area of the field having D. spicata than C. peregrina, suggesting a stronger competition with the latter. However, the increase in D. spicata density from 0-1 to >25 plants m-2 reduced fruit production by three-fold and strongly impacted vegetative growth in both fertilized and unfertilized plots. The impact of increased C. peregrina density was comparatively much lower, especially on vegetative growth, which was much higher in the area having C. peregrina. These patterns are likely due to a lower competition for N uptake with C. peregrina as this species can derive N from the atmosphere. Interestingly, the higher fruit yield in the area colonized by D. spicata occurred even in plots where the weeds were nearly absent (density of 0-1 plant m-2), revealing the influence of unidentified variables on blueberry fruit yield. We hypothesized that this difference resulted from over-optimal foliar N concentrations in the area colonized by C. peregrina as suggested by the significantly higher foliar N concentrations and by the negative correlation between foliar N concentrations and fruit yields in this area. The possibility of an influence of C. peregrina on flowering and pollination success, as well as of unidentified local site conditions is discussed. The tested N-fertilization treatments did not affect foliar N concentrations or litter decomposition rates. Overall, our results show that ammonium sulfate is very effective at increasing fruit yields but that both fruit yields and the efficiency of the N-fertilization treatment are decreased by increased D. spicata density, especially above 25 plants m-2. Although CRW did not significantly enhance fruit yields in the short term, this fertilizer may have a long-term beneficial effect.

Contrasting impacts of two weed species on lowbush blueberry fertilizer nitrogen uptake in a commercial field.

Numerous studies have speculated that lowbush blueberry (Vaccinium angustifolium) is less efficient than weed species at taking up inorganic nitrogen (N) derived from fertilizers, thus raising questions as to the effectiveness of N fertilization in commercial fields. However, competition for acquiring N as well as specific interactions between blueberry and companion weeds characterized by contrasted functional traits remain poorly documented. Here, we assessed fertilizer-derived N acquisition efficiency and biomass production in lowbush blueberry and two common weed species that have different functional traits-sweet fern (Comptonia peregrina), a N2-fixing shrub, and poverty oat grass (Danthonia spicata), a perennial grass-in a commercial blueberry field in Québec, Canada. In 2015, 15N-labelled ammonium sulfate was applied at a rate of 45 kg ha-1 to 1 m2 field plots containing lowbush blueberry and one of the two weeds present at several different density levels (0 to 25 plants m-2). In 2016, each plot was harvested to determine vegetative biomass and the percentage of fertilizer-derived N recovered (PFNR) in each species. The PFNR was higher in blueberry (24.4 ± 9.3%) than in sweet fern (13.4 ± 2.6%) and poverty oat grass (3.3 ± 2.9%). However, lowbush blueberry required about four times more root biomass than sweet fern and poverty oat grass to uptake an equivalent amount of N from ammonium sulfate. The PFNR in poverty oat grass increased with plant density (from 0.8% to 6.4% at 2-3 and >6 plants m-2, respectively), which resulted in a decrease in blueberry's PFNR (from 26.0 ± 1.4% to 8.6 ± 1.8%) and aboveground vegetative biomass production (from 152 ± 58 to 80 ± 28 g m-2). The increase in biomass production and N content in sweet fern with increasing plant density was not accompanied by an increase in PFNR (29.7 ± 8.4%), suggesting an increasing contribution of atmospherically-derived N. This mechanism (i.e., N sparing) likely explained blueberry's higher biomass production and N concentration in association with sweet fern than with poverty oat grass. Overall, our study confirms lowbush blueberry low efficiency (on a mass basis) at taking up N derived from the fertilizer as compared to weeds and reveals contrasted and complex interactions between blueberry and both weed species. Our results also suggest that the use of herbicides may not be necessary when poverty oat grass is present at a low density (<15 plants of poverty oat grass m-2) and that adding inorganic N fertilizer is counterproductive when this species is present at a high density as it takes up as much fertilizer as lowbush blueberry.

Gene Transfer with AAV9-PHP.B Rescues Hearing in a Mouse Model of Usher Syndrome 3A and Transduces Hair Cells in a Non-human Primate.

Hereditary hearing loss often results from mutation of genes expressed by cochlear hair cells. Gene addition using AAV vectors has shown some efficacy in mouse models, but clinical application requires two additional advances. First, new AAV capsids must mediate efficient transgene expression in both inner and outer hair cells of the cochlea. Second, to have the best chance of clinical translation, these new vectors must also transduce hair cells in non-human primates. Here, we show that an AAV9 capsid variant, PHP.B, produces efficient transgene expression of a GFP reporter in both inner and outer hair cells of neonatal mice. We show also that AAV9-PHP.B mediates almost complete transduction of inner and outer HCs in a non-human primate. In a mouse model of Usher syndrome type 3A deafness (gene CLRN1), we use AAV9-PHP.B encoding Clrn1 to partially rescue hearing. Thus, we have identified a vector with promise for clinical treatment of hereditary hearing disorders, and we demonstrate, for the first time, viral transduction of the inner ear of a primate with an AAV vector.

Management of Blueberry Maggot With High Temperatures.

High temperatures were investigated to manage blueberry maggot (Rhagoletis mendax Curran; Diptera: Tephritidae) in field and postharvest situations. To estimate lethal combinations of high temperatures/duration of exposure, blueberry maggot pupae were immersed in water at various temperatures during either 1 or 30 s in the laboratory. Treatments such as 70°C (1 s) or 55°C (30 s) caused 100% mortality of blueberry maggot pupae. In a lowbush blueberry field, soil temperatures at 0, 1, 2, 3, 4, and 5 cm depth were measured with thermocouples following the passage of a flamethrower attached to a tractor in the fall. While the temperatures reached up to ca. 80°C for ca. 1 s at the soil surface, they were <10°C at depths of 3, 4, and 5 cm. In field situations, the energy required to kill all pupae would be expensive to deliver with a flamethrower and this would also cause environmental concerns. An example of application concerning the use of high temperatures in a postharvest situation is discussed, notably immersion of reusable containers in hot water in compliance with Canadian Food Inspection Agency Directive D-02-04 (2015) to prevent dissemination of R. mendax in uninfested areas.

Malignant lymphoma with middle ear involvement in a Sprague-Dawley rat.

Sprague-Dawley rats are amongst the most widely used animals in biomedical research and malignant lymphoma has long been known to be a frequent neoplasm in these animals. A 9-month-old male control Sprague-Dawley rat from a toxicity study showed gelatinous material in the cranial cavity and dark, thickened cerebral meninges at necropsy. At microscopic evaluation of the temporal bone, neoplastic lymphocytes were seen invading several structures of the middle ear. The neoplastic cells appeared to extend from the marrow of the temporal bone, covered the dorsal part of the tympanic cavity wall, and surrounded and infiltrated the base of the tensor tympani muscle as well as the chorda tympani branch of the facial nerve. The lymphoma was generalized; neoplastic lymphocytes were also noted in numerous other tissues. Literature regarding neoplasms of the middle and inner ear in animals is scarce and, to our knowledge, this is the first report of a lymphoma involving the middle ear of a rat.

Middle Ear Histopathology Following Magnetic Delivery to the Cochlea of Prednisolone-loaded Iron Oxide Nanoparticles in Rats.

Delivery of therapy to the cochlea is a challenge and limits the efficacy of therapies meant to treat hearing loss, reverse tinnitus, and protect hearing from chemotherapy regimens. Magnetic injection is a technique that uses magnetic fields to inject nanoparticles from the middle ear into the cochlea, where they can then elute therapy to treat hearing disorders. To evaluate the safety of this treatment in the middle ear, 30 rats were subdivided into 6 groups and treated by single or multiple intratympanic injections of saline, prednisolone, nanoparticles, or nanoparticles loaded with prednisolone. A specially designed magnet array was used to magnetically inject the particles from the middle ear to the cochlea. Treatment began at study day 0, and animals were euthanized on study day 2, 30, or 90. Temporal bones were collected and prepared for histopathological examination. Intratympanic administration of magnetic nanoparticles and/or prednisolone resulted in minimal to mild inflammatory changes in all treated groups. The incidence and severity of the inflammatory changes observed appeared slightly increased in animals administered nanoparticles, with or without prednisolone, when compared to animals administered prednisolone alone. At study day 90, there was partial reversibility of the findings noted at study day 2 and 30. Repeat administration did not appear to cause greater inflammatory changes.

Pharmacovigilance implemented by patients: A necessity in the 21st century.

Pharmacovigilance should become a well-thought-out reflex for each of us, for any drugs taken on prescription or by self-medication. The issue of pharmacovigilance should naturally take its place in the dialogue between patients and health professionals, through advice given before medication is taken. Patients need to be told the importance of notifying any adverse drug reactions (ADRs). Pharmacovigilance would then become a post-hoc way of protecting patients. The importance of using pharmacovigilance should be promoted by emphasizing the fact that the life of a drug really starts after its marketing. This state of mind could be reinforced by a wider advertising campaign on the possibility of notifying the relevant authorities of any ADRs. Patient associations should encourage and help their members to do this. Notifying any ADRs should become a concern for all medical and paramedical staff: those who prescribe drugs, those who dispense them (pharmacists or pharmacy assistants), those who administer them (nurses), but also the consumers (patients) whose place is becoming more and more prominent.

The plant ionome revisited by the nutrient balance concept.

Tissue analysis is commonly used in ecology and agronomy to portray plant nutrient signatures. Nutrient concentration data, or ionomes, belongs to the compositional data class, i.e., multivariate data that are proportions of some whole, hence carrying important numerical properties. Statistics computed across raw or ordinary log-transformed nutrient data are intrinsically biased, hence possibly leading to wrong inferences. Our objective was to present a sound and robust approach based on a novel nutrient balance concept to classify plant ionomes. We analyzed leaf N, P, K, Ca, and Mg of two wild and six domesticated fruit species from Canada, Brazil, and New Zealand sampled during reproductive stages. Nutrient concentrations were (1) analyzed without transformation, (2) ordinary log-transformed as commonly but incorrectly applied in practice, (3) additive log-ratio (alr) transformed as surrogate to stoichiometric rules, and (4) converted to isometric log-ratios (ilr) arranged as sound nutrient balance variables. Raw concentration and ordinary log transformation both led to biased multivariate analysis due to redundancy between interacting nutrients. The alr- and ilr-transformed data provided unbiased discriminant analyses of plant ionomes, where wild and domesticated species formed distinct groups and the ionomes of species and cultivars were differentiated without numerical bias. The ilr nutrient balance concept is preferable to alr, because the ilr technique projects the most important interactions between nutrients into a convenient Euclidean space. This novel numerical approach allows rectifying historical biases and supervising phenotypic plasticity in plant nutrition studies.

Partially redundant functions of Adamts1 and Adamts4 in the perinatal development of the renal medulla.

Adamts4 encodes a widely-expressed proteinase thought to be involved in processes ranging from cartilage metabolism to ovarian follicle development. To study its physiological roles, Adamts4-null mice were created by gene targeting. Unexpectedly, these were found to be phenotypically normal, suggesting that other gene(s) may compensate for its loss. Adamts4(-/-) mice were, therefore, crossed with a strain lacking Adamts1, whose pattern of expression and substrate specificity overlap that of Adamts4. Most (>95%) Adamts1(-/-) ;Adamts4(-/-) mice died within 72 hr after birth with a marked thinning of the renal medulla. The renal defect was not observed in embryonic Adamts1(-/-) ;Adamts4(-/-) kidneys, but became apparent around birth. The few (<5%) Adamts1(-/-) ;Adamts4(-/-) animals to reach adulthood had the same renal phenotype seen in newborns. This study is thus the first to report Adamts4 expression and function in the mammalian kidney, and to demonstrate that Adamts1 and Adamts4 play redundant and essential roles in perinatal kidney development.

Middle age aggravates myocardial ischemia through surprising upholding of complex II activity, oxidative stress, and reduced coronary perfusion.

Aging compromises restoration of the cardiac mechanical function during reperfusion. We hypothesized that this was due to an ampler release of mitochondrial reactive oxygen species (ROS). This study aimed at characterising ex vivo the mitochondrial ROS release during reperfusion in isolated perfused hearts of middle-aged rats. Causes and consequences on myocardial function of the observed changes were then evaluated. The hearts of rats aged 10- or 52-week old were subjected to global ischemia followed by reperfusion. Mechanical function was monitored throughout the entire procedure. Activities of the respiratory chain complexes and the ratio of aconitase to fumarase activities were determined before ischemia and at the end of reperfusion. H(2)O(2) release was also evaluated in isolated mitochondria. During ischemia, middle-aged hearts displayed a delayed contracture, suggesting a maintained ATP production but also an increased metabolic proton production. Restoration of the mechanical function during reperfusion was however reduced in the middle-aged hearts, due to lower recovery of the coronary flow associated with higher mitochondrial oxidative stress indicated by the aconitase to fumarase ratio in the cardiac tissues. Surprisingly, activity of the respiratory chain complex II was better maintained in the hearts of middle-aged animals, probably because of an enhanced preservation of its membrane lipid environment. This can explain the higher mitochondrial oxidative stress observed in these conditions, since cardiac mitochondria produce much more H(2)O(2) when they oxidize FADH(2)-linked substrates than when they use NADH-linked substrates. In conclusion, the lower restoration of the cardiac mechanical activity during reperfusion in the middle-aged hearts was due to an impaired recovery of the coronary flow and an insufficient oxygen supply. The deterioration of the coronary perfusion was explained by an increased mitochondrial ROS release related to the preservation of complex II activity during reperfusion.

Fibrous vaginal hamartoma in a newborn calf.

A 2-day-old female Holstein calf was presented for abnormal structures protruding from the vulva at birth. A diagnosis of fibrous vaginal hamartoma was made, based on macroscopic and histologic examinations of the abnormal tissue. Management of this case involved surgical mass ablation.

First survey of fungi in hypersaline soil and water of Mono Lake area (California).

Mono Lake is a closed lake located in central California, east of the Sierra Nevada mountains. It contains dissolved carbonates, sulfates and chlorides at high concentrations. Due to its high salinity, Mono Lake was sometimes compared to the Dead Sea. However, it appears that Mono Lake water and vicinity abound with life. In this work, the fungal flora living in this extreme ecosystem was studied for the first time. Soil, tufa, water and sediment samples were also analyzed for their mineral and salt composition. Results showed that water was particularly rich in sodium, potassium, phosphorus and boron. Soil and sediments contained very high levels of calcium and magnesium, but also barium, boron and strontium. Sodium, phosphorus and iron levels varied in a large extent from one to another sample. Neutral to very alkaline pH were recorded. Water samples were found sterile in the conditions chosen for fungi isolation, while sediment, soil and tufa samples led to the isolation of a total of 67 fungal species (from 23 samples), belonging to various taxonomic groups. From our results no clear effects of the chemical parameters of the samples were observed on fungal life apart from the pH. The methods chosen did not allow the isolation of extremely halotolerant species. We isolated in this work a series of ubiquitous species, suggesting that a selection of resistant and/or adaptable strains of some common species could have occurred. Depending on the medium and the temperature of isolation, it can be hypothesized that some species were present as dormant structures, while some others, isolated at pH 8 on a medium enriched in Na and Ca, could be in a growing form adapted to alkaline and saline conditions. This work contributes to a better knowledge of the mycobiota present in the Mono Lake's ecosystem.