Physiological and biochemical differences among Ulmus minor genotypes showing a gradient of resistance to Dutch elm disease

Dutch elm disease (DED) spread across Europe and North America in the 20th century killing most natural elm populations. Today, breeding programmes aim at identifying, propagating and studying elm clones resistant to DED. Here, we have compared the physiology and biochemistry of six genotypes of Ulmus minor of variable DED resistance. Leaf gas exchange, water potential, stem hydraulic conductivity and biochemical status were studied in 5‐year‐old trees of AB‐AM2.4, M‐DV2.3, M‐DV2 × M‐CC1.5 and M‐DV1 and 6‐year‐old trees of VA‐AP38 and BU‐FL7 before and after inoculation with Ophiostoma novo‐ulmi. Leaf water potential and net photosynthesis rates declined, while the percentage loss of hydraulic conductivity (PLC) increased after the inoculation in susceptible trees. By the 21st day, leaf predawn and midday water potential, stomatal conductance to water vapour and net photosynthesis rates were lower, and PLC was higher in trees of susceptible (S) genotypes inoculated with the pathogen than in control trees inoculated with water, whereas no significant treatment effect was observed on these variables in the resistant (R) genotypes. Fourier transform infrared spectroscopy analyses revealed a different biochemical profile for branches of R and S clones. R clones showed higher absorption peaks that could be assigned to phenolic compounds, saturated hydrocarbons, cellulose and hemicellulose than S clones. The differences were more marked at the end of the experiment than at the beginning, suggesting that R and S clones responded differently to the inevitable wounding from inoculation and repeated sampling over the experimental course. We hypothesize that a weak activation of the defence system in response to experimental wounding can contribute to the susceptibility of some genotypes to O. novo‐ulmi. In turn, the decline in shoot hydraulic conductivity and leaf carbon uptake caused by the infection further exacerbates tree susceptibility to the fungus.     

Laboratory and on‐farm evaluation of low‐cost salt mixtures for use during salinity acclimation and the nursery phase of Pacific white shrimp (Litopenaeus vannamei)

The current study evaluates the efficacy of a low‐cost salt mixture (LCSM) to replace expensive reconstituted sea salt (RSS) in the salinity acclimation and nursery phase of Pacific white shrimp under laboratory and farm conditions. LCSM was formulated to yield sodium, potassium, calcium and magnesium concentrations closely comparable to that of diluted seawater. Laboratory‐based nursery trials were conducted at 2, 6 and 15 g/L salinities, incrementally replacing RSS with LCSM (25%, 50%, 75% and 100%) at four replicates per treatment. Thirty postlarvae were reared for 7 days in 24‐L aquaria during the 2 and 6 g/L trials, while the nursery trial for 15 g/L salinity was conducted for 21 days with 400 postlarvae stocked in 150‐L tanks. On‐farm evaluation of LCSM was carried out in two tank‐based systems installed on levees adjacent to shrimp production ponds. RSS was incrementally replaced with LCSM (0%, 50%, 75% and 100%) and 100 postlarvae stocked into each 800‐L tank. Salinity acclimation was done from 30 g/L to 6 or 1.5 g/L within 2–3 days by pumping water from adjacent shrimp production ponds. Following salinity acclimation, the S4 system maintained flow‐through at 1.5 g/L, while N10 system was maintained static at 6 g/L salinity. At the conclusion, no significant differences were observed for either survival or growth of shrimp postlarvae between RSS and LCSM treatments at all salinities examined. Results reflect the potential use of LCSM to replace RSS, which could be an excellent solution to bring down the cost of production in inland shrimp aquaculture.     

The Alleviating Effect of Elevated CO2 on Heat Stress Susceptibility of Two Wheat (Triticum aestivum L.) Cultivars

This study analysed the alleviating effect of elevated CO₂ on stress‐induced decreases in photosynthesis and changes in carbohydrate metabolism in two wheat cultivars (Triticum aestivum L.) of different origin. The plants were grown in ambient (400 μl l^?1) and elevated (800 μl l^?1) CO₂ with a day/night temperature of 15/10 °C. At the growth stages of tillering, booting and anthesis, the plants were subjected to heat stress of 40 °C for three continuous days. Photosynthetic parameters, maximum quantum efficiency of photosystem II (PSII) photochemistry (F_v/F_m) and contents of pigments and carbohydrates in leaves were analysed before and during the stress treatments as well as after 1 day of recovery. Heat stress reduced P_N and F_v/F_m in both wheat cultivars, but plants grown in elevated CO₂ maintained higher P_N and F_v/F_m in comparison with plants grown in ambient CO₂. Heat stress reduced leaf chlorophyll contents and increased leaf sucrose contents in both cultivars grown at ambient and elevated CO₂. The content of hexoses in the leaves increased mainly in the tolerant cultivar in response to the combination of elevated CO₂ and heat stress. The results show that heat stress tolerance in wheat is related to cultivar origin, the phenological stage of the plants and can be alleviated by elevated CO₂. This confirms the complex interrelation between environmental factors and genotypic traits that influence crop performance under various climatic stresses.     

The use of intrathecal analgesia and contrast radiography as preoperative diagnostic methods for digital flexor tendon sheath pathology

Reasons for performing study: The sensitivity of ultrasonography for the diagnosis of manica flexoria (MF) tears within the digital flexor tendon sheath (DFTS) is lower than for diagnosis of marginal tears of the deep digital flexor tendon (DDFT). Additional diagnostic tools would assist in appropriate decision making for either conservative or surgical management. Objectives: To evaluate the improvement in lameness of horses with MF or DDFT tears following intrathecal analgesia and to assess the sensitivity and specificity of contrast radiography for the diagnosis of these tears. Methods: The case records of horses presented to a referral clinic over a 7‐year period that underwent intrathecal diagnostic analgesia, or intrathecal analgesia and contrast radiography, of the DFTS with subsequent tenoscopy were examined. Results: Fifty‐three limbs had intrathecal diagnostic analgesia performed and 23 contrast tenograms were assessed in horses undergoing DFTS tenoscopy. Horses with DDFT tears were significantly more likely to respond positively to intrathecal diagnostic analgesia than those with MF tears (P = 0.02). Using contrast radiography, tears of the MF were predicted with an overall sensitivity of 96% and specificity of 80%; marginal tears of the DDFT were predicted with an overall sensitivity of 57% and specificity of 84%. Conclusions: The results of intrathecal analgesia of the DFTS in combination with contrast radiography have a high sensitivity for predicting MF tears. The sensitivity of contrast radiography for predicting tears of the DDFT is lower but the specificity remains high. Potential relevance: Contrast radiography performed at the same time as intrathecal analgesia provides useful information regarding the presence of MF tears and DDFT tears, which can assist in the decision of whether to manage the lameness conservatively or with tenoscopic evaluation.     

Feed preference and growth response of juvenile Litopenaeus vannamei to supplementation of marine chemoattractants in a fishmeal‐challenged diet

This study compared the feed preference and growth response of Litopenaeus vannamei to chemoattractants. A diet with 3% fishmeal was supplemented with either 3% salmon meal (POS), 3% soy protein concentrate (NEG), 3% krill meal (KRM), 3% squid meal (SQM), 3% shrimp head meal (SHM), 3% shrimp meal (SM), 3% squid liver meal (SLM), or 5% liquid sardine hydrolysate (SAH). Shrimp with a body weight (BW) of 0.99 ± 0.08 g were stocked at 100 animals/m² in 56 tanks of 1 m³ and fed 10 times daily for 74 days. Feed preference was evaluated by feeding shrimp of 10.87 ± 1.82 g in excess twice a day for 10 days in two separate feeding trays allocated in 50 tanks of 0.5 m³. Survival reached 93.3 ± 5.80% and was unaffected by supplementation. Final BW was the highest for shrimp fed the KRM‐supplemented diet (11.97 ± 0.93 g), followed by POS (11.11 ± 0.77 g) and SQM (11.01 ± 1.17 g). Diets SHM, SM, SLM, and NEG showed a lower shrimp BW than POS, but were not statistically different among them. Shrimp fed the SAH diet achieved the lowest BW (10.06 ± 1.02 g). The highest gained yield was obtained with diets KRM and POS. No statistical difference was observed in shrimp yield among other diets. The lowest feed conversion ratio (FCR) was achieved with shrimp fed KRM (1.31 ± 0.05) when compared to diets SHM (1.47 ± 0.05), SAH (1.47 ± 0.07), and SLM (1.45 ± 0.17). Two‐by‐two comparisons indicated that shrimp preferred SHM and KRM, except when these were compared to SQM and SLM. No difference in feed preference was found between diets with SQM and SLM. SAH was the least preferred raw material in all comparisons. Results indicated that KRM acts as a powerful feeding effector and growth enhancer in fishmeal‐challenged diets for whiteleg shrimp. A dietary supplementation with 3% KRM is more effective than the same dose of any other chemoattractant evaluated.     

In vitro effect of levamisole on the cell viability,phagocytosis and respiratory burst of Barbel chub (Squaliobarbus curriculus) macrophages

The present study was to determine in vitro effect of levamisole on the immune functions of Barbel chub (Squaliobarbus curriculus), head–kidney‐derived (HKM), spleen‐derived (SM) and peripheral blood monocyte‐derived macrophage (PBM). Macrophages were incubated with levamisole 10³, 10¹, 10⁻¹ and 10⁻³ ng mL⁻¹ to assay the cell viability, respiratory burst and phagocytosis. The results showed that macrophages treated with levamisole 10⁻³ ng mL⁻¹ gave a maximum respiratory burst response, whereas levamisole 10³ ng mL⁻¹ had no effect. Phagocytosis activity of the macrophages treated with levamisole enhanced significantly when compared to control, maximum response being at 10⁻³ ng mL⁻¹. While using the methylthiazoletetrazolium method to measure the cell activity for 12, 24, 36, 48, 60 h, there was no significant role on proliferation and the cell viability began to decline after 24 h. In conclusion, levamisole is a potent enhancer of Barbel chub macrophage activity with low concentration.     

Influence of the root endophyte Piriformospora indica on the plant water relations,gas exchange and growth of Chenopodium quinoa at limited water availability

This study aimed to evaluate the ability of Piriformospora indica to colonize the root of Chenopodium quinoa and to verify whether this endosymbiont can improve the growth, performance and drought resistance of this species. The study delivered, for the first time, evidence for successful colonization of P. indica in quinoa. Hence, pot experiment was conducted in the greenhouse, where inoculated and non‐inoculated plants were subjected to ample (40%–50% WHC) and deficit (15%–20%WHC) irrigation treatments. Drought adversely influenced the plant growth, leading to decline the total plant biomass by 74%. This was linked to an impaired photosynthetic activity (caused by lower g_s and C_i/C_a ratio; stomatal limitation of photosynthesis) and a higher risk of ROS production (enhanced ETR/A_gross ratio). P. indica colonization improved quinoa plant growth, with total biomass increased by 8% (controls) and 76% (drought‐stressed plants), confirming the growth‐promoting activity of P. indica. Fungal colonization seems to diminish drought‐induced growth hindrance, likely, through an improved water balance, reflected by the higher leaf ψ_w and g_s. Additionally, stomatal limitation of photosynthesis was alleviated (indicated by enhanced C_i/C_a ratio and A_net), so that the threat of oxidative stress was minimized (decreased ETR/A_gross). These results infer that symbiosis with P. indica could negate some of the detrimental effects of drought on quinoa growth, a highly desired feature, in particular at low water availability.     

Genetic diversity and relatedness estimates for captive barramundi (Lates calcarifer,Bloch) broodstock informs efforts to form a base population for selective breeding

Aquaculture of barramundi or Asian seabass (Lates calcarifer) is growing in both Australia and Southeast Asia and there is substantial interest to improve production efficiency through selective breeding. The establishment of a large and genetically diverse base population is a prerequisite for a sustainable and long‐term productive breeding program. Before selective breeding programs can begin for Australian barramundi it is important to assess the overall genetic diversity of current captive broodstock populations. To address this question, 407 captive barramundi broodstock from eight separate Australian broodstock populations were genotyped using 16 polymorphic microsatellite DNA markers. A Bayesian STRUCTURE analysis indicated that captive Australian broodstock are broadly divided into two genetic stocks. Multivariate analysis between broodstock individuals and pairwise F_ST between broodstock populations also supported the existence of two stocks. Comparisons with data obtained from natural stocks suggested that hatchery individuals were either sourced from the two stocks or represented an admixture between them. Genetic diversity was low within each broodstock population (allelic richness ranged from 2.67 to 3.42 and heterozygosity ranged from 0.453 to 0.537) and relatedness estimates within hatcheries were generally low (average r was equal to 0.141). We recommend sourcing captive individuals according to high levels of neutral genetic diversity and low levels of relatedness for the establishment of a base population. We also make recommendations about including genetically diverse wild individuals.     

Osteological development and anomalies in larval stage of hatchery‐reared yellow catfish Pelteobagrus fulvidraco

Comprehensive knowledge of osteological development of fish not only provides means for understanding its functional development, but also allows early detection of skeletal anomalies. The present study was conducted to determine osteological development and occurrence of anomalies from the first to the 40th day after hatching (DAH) for yellow catfish Pelteobagrus fulvidraco, a commercially important species in China. Vertebral ontogeny started with formation of anterior neural arches at 3 DAH, and completed at 6 DAH. Vertebral centra started to develop at 4 DAH and ossification in all centra was visible at 15 DAH. Caudal fin appeared at 2 DAH and ossification was visible at 26 DAH. The onset of dorsal and anal fins appeared at 4 DAH and their ossifications were visible at 20 DAH. Pectoral fins were present before first feeding and formed entirely at 9 DAH, and their ossifications were visible at 21 DAH. Pelvic fins appeared at 9 DAH, and formed completely at 21 DAH. The ossification of pelvic fins was not finished until the end of the experiment (40 DAH). 24 types of skeletal anomalies were observed. About 20% of individuals showed at least one anomaly. Haemal vertebrae anomalies occurred at the highest frequency, followed by pre‐haemal, caudal vertebrae and anal fin anomalies, and caudal fin had the lowest anomalies frequency. For the first time, our study determined osteological development and anomalies incidence in larval yellow catfish, which help further investigations into rearing conditions leading to appearance of these anomalies to prevent their incidence.