Multilocus sequence typing detects new Piscirickettsia salmonis hybrid genogroup in Chilean fish farms: Evidence for genetic diversity and population structure

Piscirickettsia salmonisis the causative bacterial pathogen of piscirickettsiosis, a salmonid disease that causes notable mortalities in the worldwide aquaculture industry. Published research describes the phenotypic traits, virulence factors, pathogenicity and antibiotic‐resistance potential for various P. salmonisstrains. However, evolutionary and genetic information is scarce for P. salmonis. The present study used multilocus sequence typing (MLST) to gain insight into the population structure and evolution of P. salmonis. Forty‐two Chilean P. salmonisisolates, as well as the type strain LF‐89^T, were recovered from diseased Salmo salar, Oncorhynchus kisutchand Oncorhynchus mykissfrom two Chilean Regions. MLST assessed the loci sequences of dnaK, efp, fumC, glyA, murG, rpoD and trpB. Bioinformatics analyses established the genetic diversity among P. salmonis isolates (H = 0.5810). A total of 23 sequence types (ST) were identified, 53.48% of which were represented by ST1, ST5 and ST2. Population structure analysis through polymorphism patterns showed few polymorphic sites (218 nucleotides from 4,010 bp), while dN/dS ratio analysis indicated purifying selection for dnaK, epf, fumC, murG, and rpoD but neutral selection for the trpB loci. The standardized index of association indicated strong linkage disequilibrium, suggesting clonal population structure. However, recombination events were detected in a group of seven isolates. Findings included genogroups homologous to the LF‐89^T and EM‐90 strains, as well as a seven‐isolate hybrid genogroup recovered from both assessed regions (three O. mykiss and four S. salar isolates). The presented MLST scheme has comparative potential, with promising applications in studying distinct P. salmonis isolates (e.g., from different hosts, farms, geographical areas) and in understanding the epidemiology of this pathogen.     

Performance of a photo‐heterotrophic,hypersaline system for intensive cultivation of white leg shrimp (Litopenaeus vannamei) with minimal water replacement in lined ponds using a stochastic approach

The aim of the study was to evaluate the performance of Litopenaeus vannamei in an intensive photo‐heterotrophic hypersaline system with minimal seawater replacement, and establish relationships between parameters of a stochastic production model and relevant water quality variables. Six experimental 1000 m² lined ponds were stocked at a density of 120 shrimp m^?2 for a 105‐day trial. Salinity increased from 37 to 45 ± 2 g/L, and the water level was maintained with the weekly addition of filtered seawater, equivalent to 1.6% per day. The stochastic model predicted that, at harvest, there is 95% confidence that the system produces between 12.1 and 14.7 t/ha with a mean final individual weight of 13.1 g and a mean survival of 84.2%. Sensitivity analyses showed that dissolved oxygen and individual final weight of shrimp were the main variables influencing yield variance. Nitrogenous compounds were maintained between optimal cultivation levels (NH₃–NH₄⁺ = 0.73 ± 0.43 mg/L, N–NO₂^? = 0.09 ± 0.05 mg/L, N–NO₃^? = 3.22 ± 0.11 mg/L). Heterotrophic bacteria (6.6 ± 3.4 × 10⁵ CFU/ml) and chlorophyll‐α concentration (108.5 ± 80.2 μg/L) showed a similar development pattern, indicating a strong relationship between bacteria and microalgae during cultivation. Vibrio spp. concentrations were low (1.24 ± 1.42 × 10³ CFU/ml). It was shown that the photo‐heterotrophic system could be used in hypersaline conditions, typical of semi‐arid regions, to consistently produce between 12.1 and 14.7 t/ha in 15 weeks.     

Influence of High Molecular Weight Glutenins on Viscoelastic Properties of Intact Wheat Kernel and Relation to Functional Properties of Wheat Dough

High and low molecular weight glutenin subunits (HMW‐GS and LMW‐GS, respectively) are the main factors determining the viscoelastic properties of wheat dough. The mechanical and viscoelastic properties of 29 samples of wheat kernels differing in HMW‐GS were evaluated with load‐compression tests. Samples were grouped by genotypes differing in HMW‐GS composition (allelic variants: Glu‐A1: null, 1, 2*; Glu‐B1: 7, 7+8, 7+9, 13+16, and 17+18; Glu‐D1: 5+10, 2+12). Groups representing Glu‐A1 1 and 2*; Glu‐B1 7, 7+9 and 17+18; and Glu‐D1 5+10 generally possessed hard grain and showed the largest kernel elasticity values, while those representing subunits Glu‐A1 null; Glu‐B1 7+8; and Glu‐D1 2+12 had soft kernels and showed lower elastic work values. Genotypes possessing HMW‐GS 1, 17+18 and 5+10 gave large SDS‐sedimentation values and better dough viscoelastic properties than those with allelels: null, 7+8, and 2+12. Kernel hardness showed significant correlation with the dough‐strength‐related parameters: SDS‐sedimentation; dough mixing time; and the alveographic parameters, W and P. There was a negative correlation between kernel plastic work and dough mixing time and the dough tenacity/extensibility parameters, P/L. The significant relationship between sedimentation tests and kernel elastic work seems to indicate that elastic work is related to genotype (protein composition). The general tendency was that higher values in kernel elastic work and size corresponded to better dough rheological quality. Mechanical properties of the kernel were significantly related to the elastic behavior measured in a single wheat kernel. The use of the compression test on individual kernels is easy, rapid and nondestructive and therefore seems to show potential use as a rapid tool in breeding to improve wheat quality.     

Evaluation of Degree of Elasticity and Other Mechanical Properties of Wheat Kernels

Three samples were selected representing bread, soft, and durum wheat. Uniaxial compression and stress relaxation tests were performed on wheat kernels. Force‐deformation curves from intact wheat grain typically exhibited at least two points of inflection (PI) at ≈0.1 and 0.2 mm displacement. The first PI is related to the mechanical properties of all the bran layers. The second PI (0.2 mm) seems to be the endosperm boundary near the aleurone layer. These structures had higher degree of elasticity (DE) compared to the inner endosperm (0.5–0.6 mm). Besides wheat class and specific structures of the caryopsis, moisture content is a prominent factor affecting the mechanical strength of kernels. Stress relaxation tests show that bread wheat kernels with 69.2% DE at 13% moisture decreased to 31.6% DE with additional 6% moisture content. Soft wheat kernels DE of 61.0% at 13% moisture decreased to 22.7% at 19.7% moisture. Stress relaxation revealed pronounced time‐dependence. However, the differences of stress values at 120–180 sec were not significant in all wheat classes and moisture contents evaluated. The stress values after 120 sec might be attributed to the elastic deformation of the kernels.     

玉米花粉寿命及有效控制花粉飘逸的隔离距离研究

花粉是玉米基因飘逸的重要载体。研究了花粉活力的持续时间和有效控制基因飘逸的隔离距离。在花药刚开裂时采集花粉,置于薄膜层上,放在室外自然条件下保存,每隔一定时间对采集的花粉进行目测和授粉．通过对花粉的目测评判和计算结实率评估花粉活力。试验花粉源区域4000m^2,在其周围不同距离的地方种植若干玉米小区,每个小区面积为12．8m^2,以测定隔离距离的有效性。作为花粉源的玉米材料包含可用于测定花粉飘逸的叶色标记或种子标记。由于大气压不同,花粉可在花药开裂后1～2小时内保持活力。假定花粉以当地下午平均最大风速直线传播,并可在2小时内保持花粉活力,且忽略花粉沉降率因素,理论上讲,具有活力的花粉可以传播的最大距离为32km。试验结果是在离花粉源100m发生了一定数量的异花授粉,离花粉源200m处有个别异花授粉发生。离花粉源300m处没有异花授粉发生。试验结果与玉米花粉不耐干燥并具高沉降速率的结论一致。试验结果表明距离隔离是控制玉米因授粉而发生基因飘逸的有效方法。     

Study of Viscoelastic Properties of Wheat Kernels Using Compression Load Method

The method to measure hardness and other viscoelastic properties of intact wheat kernels is presented. Wheat with 9.3% moisture showed high elastic behavior compared with wheat tempered at 22.5% moisture that showed a plastic behavior. Load‐deformation curves showed that bread wheat behaves as a more plastic material than durum wheat, which is a more elastic material. Yield point of all the wheat samples was ≈18.5 N, independent of wheat type and moisture content. The height of the wheat kernel increased linearly, and the compression area increased exponentially, with increasing moisture content. The modulus of elasticity of wheat ranged from 99.2 MPa for 22.5% moisture content to 394.8 MPa for 9.3% moisture content. Young's modulus range for soft wheat such as Salamanca, Saturno, and Cortazar cultivars was 232.2–308.5 MPa, as compared with Rayón bread wheat at 321.5 MPa and the Altar, Sofía, and Rafi cultivars of durum wheat that had elastic moduli of 438.7–485.8 MPa. The compression force and final stress decreased from 69.9 N and 40.1 MPa in soft wheat to 90 N and 78.9 MPa in durum, respectively. Total work range was 14.7 MPa/sec in soft wheat to 19.7 MPa/sec for durum wheat and, as expected, was higher in the durum and bread wheat than in soft wheat. The plastic part ranged from 2.4 MPa/sec in soft wheat to 0.6 MPa/sec in durum wheat.     

An approach to the evaluation and management of natural carbon sinks: From plant species to urban green systems

The maintenance and improvement of current natural carbon sinks, as well as the creation of others, are presently considered among the main strategies by which to address global climate change, and are pursued in parallel to the implementation of policies of carbon emission reduction. To date, this approach has been applied mainly to forest areas, obtaining relevant results that help to identify areas of weakness in the strategy and to design appropriate management measures. Nevertheless, under the current situation, and at territorial level, urban areas and infrastructures, in which urban green systems play a significant role, present opportunities for contributing towards carbon sink potential. Various studies conducted by our research group have emphasized the role of urban green systems as natural carbon sinks, including research conducted under initiatives from European authorities (“Climeport project/Mediterranean Ports’ Contribution to Climate Change”), regional government (Andalusian Strategy against Global Climate Change) and Andalusian universities (Andalusian University Rectors Climate Change Pact). Through these projects, we have assessed entire systems, identifying the species of urban trees and shrubs with the greatest sink potential under different climate scenarios, proposing garden management schemes, and providing a basis for the creation, design, management and improvement of natural carbon sinks in the cities.     

Analysis of the Mycosporine-Like Amino Acid (MAA) Pattern of the Salt Marsh Red Alga Bostrychia scorpioides

This study presents the validation of a high-performance liquid chromatography diode array detector (HPLC-DAD) method for the determination of different mycosporine-like amino acids (MAAs) in the red alga Bostrychia scorpioides. The investigated MAAs, named bostrychines, have only been found in this specific species so far. The developed HPLC-DAD method was successfully applied for the quantification of the major MAAs in Bostrychia scorpioides extracts, collected from four different countries in Europe showing only minor differences between the investigated samples. In the past, several Bostrychia spp. have been reported to include cryptic species, and in some cases such as B. calliptera, B. simpliciuscula, and B. moritziana, the polyphyly was supported by differences in their MAA composition. The uniformity in the MAA composition of the investigated B. scorpioides samples is in agreement with the reported monophyly of this Bostrychia sp.     

Effects of methadone, alone or in combination with acepromazine or xylazine, on sedation and physiologic values in dogs

ObjectiveTo evaluate the effects of methadone, administered alone or in combination with acepromazine or xylazine, on sedation and on physiologic values in dogs.Study designRandomized cross-over design.AnimalsSix adult healthy mixed-breed dogs weighing 13.5 ± 4.9 kg.MethodsDogs were injected intramuscularly with physiologic saline (Control), or methadone (0.5mg kg⁻¹) or acepromazine (0.1 mg kg⁻¹) or xylazine (1.0 mg kg⁻¹), or acepromazine (0.05 mg kg⁻¹) plus methadone (0.5 mg kg⁻¹) or xylazine (0.5 mg kg⁻¹) plus methadone (0.5 mg kg⁻¹) in a randomized cross-over design, with at least 1-week intervals. Sedation, pulse rate, indirect systolic arterial pressure, respiratory rate (RR), body temperature and pedal withdrawal reflex were evaluated before and at 15-minute intervals for 90 minutes after treatment.ResultsSedation was greater in dogs receiving xylazine alone, xylazine plus methadone and acepromazine plus methadone. Peak sedative effect occurred within 30 minutes of treatment administration. Pulse rate was lower in dogs that received xylazine either alone or with methadone during most of the study. Systolic arterial pressure decreased only in dogs receiving acepromazine alone. When methadone was administered alone, RR was higher than in other treatments during most of the study and a high prevalence of panting was observed. In all treatments body temperature decreased, this effect being more pronounced in dogs receiving methadone alone or in combination with acepromazine. Pedal withdrawal reflex was absent in four dogs receiving methadone plus xylazine but not in any dog in the remaining treatments.Conclusions and clinical relevanceMethadone alone produces mild sedation and a high prevalence of panting. Greater sedation was achieved when methadone was used in combination with acepromazine or xylazine. The combination xylazine–methadone appears to result in better analgesia than xylazine administered alone. Both combinations of methadone/sedative were considered effective for premedication in dogs.     

Recent spreading of a divergent canine parvovirus type 2a (CPV-2a) strain in a CPV-2c homogenous population

Canine parvovirus type 2 (CPV-2), which causes acute hemorrhagic enteritis in dogs, is comprised of three antigenic variants (2a, 2b, and 2c) that are distributed worldwide with different frequencies. Variant prevalence was analyzed in 150 CPV-2-positive samples collected from the Uruguayan dog population in 2007-2010. Samples were analyzed with polymerase chain reaction, restriction fragment length polymorphism, and sequencing of the coding region for the largest and most variable loop of the VP2 capsid protein. CPV-2c was the only strain detected from 2007 to 2009. Uruguayan CPV-2c showed high homogeneity in both nucleotide and amino acid sequences, indicating a low level of genetic variability. In 2010, an unexpected epidemiological change occurred in Uruguay as a consequence of the appearance of a novel CPV-2a strain. This variant rapidly spread through the Uruguayan dog population and was detected in 20 of the 52 cases (38%) analyzed in 2010. CPV-2a sequences were identical in all field viruses analyzed, and in addition to the characteristic 426Asn residue, the sequences showed amino acid substitutions (267Tyr, 324Ile, and 440Ala) not observed in the Uruguayan CPV-2c. These data and the first detection in April 2010 suggest that the CPV-2a variant recently emerged in Uruguay and underwent clonal expansion. This observation is the first case in which a CPV-2a variant increased its frequency in a dog population where CPV-2c was prevalent. Our results emphasize the dynamic changes in CPV variants and highlight the importance of ongoing surveillance programs to provide a better understanding of virus epidemiology.