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.     

Effect of dietary carbohydrate-to-lipid ratios on growth, lipid deposition and metabolic hepatic enzymes in juvenile Senegalese sole (Solea senegalensis, Kaup)

A study was undertaken to determine the effect of various dietary carbohydrate‐to‐lipid ratios on growth performance, whole‐body composition and tissue lipid content in Senegalese sole (Solea senegalensis) juveniles. Data on the dietary regulation of key hepatic enzymes of the lipogenic and glycolytic pathways (glucose‐6‐phosphate dehydrogenase, G6PD; malic enzyme, ME; fatty acid synthetase, FAS; pyruvate kinase, PK and glucokinase, GK) were also generated. Four isonitrogenous (crude protein: 52% dry matter (DM)) diets were formulated to contain one of two lipid levels (11% and 21% DM). Within each dietary lipid level, the nature of the carbohydrate fraction (raw or extruded peas) was varied. Triplicate groups of 54 sole (initial body weight: 23.6±1.2 g) were grown in recirculated seawater over 67 days. Fish were fed using automated feeders. At the end of the study, whole‐body, liver, viscera and muscle samples were withdrawn for analyses. During the experimental period, the mean fish weight about doubled in all treatments. No significant differences were found in growth performance (ranging from 1.1% to 1.4% body weight day^?1) among dietary treatments. High‐fat diets increased whole‐body fat content. Similarly, daily fat gain ranged from 0.54 to 0.78 g kg^?1 day^?1 and highest values were found in fish fed high‐lipid diets. Dietary treatments also affected tissue lipid content (liver, viscera and muscle), with highest values in fish fed high‐fat diets. The nature of dietary carbohydrates had little influence on performance criteria, but affected tissue lipid deposition. The activities of G6PD, ME and FAS were depressed by elevated levels of dietary lipid, confirming the inhibitory effect of dietary fats on lipid biosynthesis. At both dietary lipid levels, ME and FAS activities were little affected by dietary carbohydrate. Activities of PK and GK were not affected by the starch level of the diets. In Senegalese sole juveniles, the lipogenic pathway is more susceptible to modulation by dietary means (particularly through lipid intake) than the glycolytic pathway.     

Toxicity of aqueous aluminium to the ectoparasitic monogenean Gyrodactylus salaris

Both laboratory and field experiments have demonstrated that aqueous aluminium can act as a paraciticide to the monogenean Gyrodactylus salaris Malmberg, 1957 infecting freshwater salmonids. However, the reproductive conditions of gyrodactylids surviving to cessation of an aluminium exposure is unknown. Therefore, Atlantic salmon (Salmo salar L.) parr infected with G. salaris was experimentally exposed to elevated concentrations of aqueous aluminium for more than 1 month. During this period, the infection increased the first week before it peaked and started to steadily decline approaching elimination. When almost all parasite specimens were eliminated, the water quality was adjusted to normal aluminium-poor freshwater. During the next 3 weeks, the surviving G. salaris were shown to clearly resume their reproduction. The experiment demonstrates the potential of the gyrodactylids to reproduce after cessation of an aluminium exposure. Thus, the as-yet unknown mechanism behind the toxic effects of aluminium on G. salaris survival and reproduction might to some extent be reversible. The present study gives further support for this metal as a parasiticide to G. salaris but demonstrates at the same time that in order to eliminate the parasites totally, the aluminium treatment must eradicate all G. salaris from the skin of the infected hosts. Furthermore, the potential development of resistance to aluminium should also be studied.     

Morphological development and allometric growth of yellowtail kingfish Seriola lalandi V. larvae under culture conditions

Morphological development and allometric growth patterns of Seriola lalandi larvae were assessed to characterize normal growth patterns under culture conditions. Early ontogenetic stages of yellowtail kingfish exhibited an exponential growth in terms of standard length as a function of age. Five development stages were characterized from hatching to the juvenile stage: larval stage I (0–2 days post hatch, dph) with endogenous feeding, characterized by a small yolk sac, unpigmented eyes, primordial finfold surrounding the body and a closed mouth; larval stage II (2–15 dph) characterized by mouth opening, complete pigmentation of eyes and the beginning of the exogenous feeding; subsequently, in the larval stage III (15–25 dph) the posterior tip of notochord of the larvae bended upward and the first rays appeared in fins, concomitant with a change in swimming behaviour; thereafter, larval stage IV (post‐flexion stage; 25–30 dph) began when larvae resembled in morphology to a juvenile organism; however, caudal and dorsal fins were not completely development. Lastly, the juvenile stage was reached 30 dph characterized by a morphology and fin structures similar to those of the adults. Growth and development of structures and organs associated with vital functions such as feeding, sensorial and breathing systems seemed to be more critical previous to 23 dph, which was reflected with a positive allometric growth of head and eyes during this period. The results from this study can be used as a tool‐guide to assess normal development in larval research with S. lalandi to improve existing rearing protocols in hatchery production.     

Extrinsic and intrinsic factors influence daily activity of a Mediterranean cyprinid

Factors shaping individual behavior remain poorly understood for European cyprinids. We examined the effects of extrinsic (temperature and year) and intrinsic factors (body size and shoal size) on the daily activity of chub Squalius torgalensis, a critically endangered cyprinid found in the Torgal stream, Portugal. We quantified chub behaviors while snorkeling, using instantaneous sampling of focal individuals, in spring 2009 and 2010. We used Principal Component Analysis to describe individual behavior and assessed competing models relating individual behavior to explanatory variables using Akaike's Information Criterion. Year had a strong influence on chub behavior, with individuals spending more time drift foraging and orienting in 2009, whereas in 2010 they spent more time benthic foraging and cruising. Shoal size did not influence behaviors in 2009, but in 2010 individuals in small shoals foraged benthically and searched with greater frequency than those in large shoals. Individuals increased cruising and decreased searching as they grew in length, but the opposite trend was observed as temperature increased. Our results indicate that both extrinsic and intrinsic factors influence daily activity of chub, with individual behaviors varying between years, and depending upon shoal size, body size and temperature. This suggests that chub display high levels of individual behavioral flexibility, which may be important for fish inhabiting harsh and highly variable Mediterranean streams. Conservation efforts for this species will benefit by integrating behavioral information into management strategies.     

Effect of 4-amino-6-methyl-3-phenylamino-1,2,4-triazin-5(4H)-one on the integrity of chloroplasts from barley leaves

4-Amino-6-methyl-3-phenylamino-1,2,4-triazin-5(4H)-one alters the integrity of chloroplasts in barley (Hordeum vulgare L. var. Hassan) leaves as judged by their sedimentation profiles in linear gradients of sucrose. This effect may be related to the inhibition caused by 4-amino-6-methyl-3-phenylamino-1,2,4-triazine-5(4H)-one of both ascorbate peroxidase and superoxide dismutase activities, and could thus be explained by an accumulation of toxic reduced-oxygen species in illuminated chloroplasts.     

Microvascular development and growth of uterine tissue during the estrous cycle in mares

OBJECTIVE: To document uterine growth and microvascular development in the endometrium of uteri with differing degrees of fibrosis as well as uterine growth throughout the estrous cycle of mares. ANIMALS: 30 mares. PROCEDURE: Uterine tissue was obtained during the breeding season from a slaughter facility. Stage of estrous cycle of the mares was assessed on the basis of ovarian structures and plasma progesterone concentrations. Endometrium was characterized by use of light microscopy, and blood vessel walls were marked by histochemical techniques. Microvascular development was evaluated by a computerized image analysis system. Growth of uterine tissue was based on cellular content of DNA and RNA, RNA:DNA, and protein:DNA. RESULTS: Significant differences in vascular density were not observed in the endometrium of uteri obtained from mares euthanatized during the follicular or luteal phase of the estrous cycle, regardless of whether endometrial classification of degree of fibrosis was considered. There was a 3-fold increase in amount of DNA and RNA of endometrial cells in the follicular phase when compared to myometrium. Hypertrophy of endometrial tissue during the luteal phase was reflected by a significant increase in cell protein content and protein:DNA. CONCLUSIONS AND CLINICAL RELEVANCE: Endometrial growth of vascular tissues during the estrous cycle may be coordinated with development of nonvascular tissue. Estrogen and progesterone may play a role in regulation of uterine growth and angiogenesis.     

Crude protein fractions in common vetch (Vicia sativa L.) fresh forage during pod filling

Crude protein (CP) of forages can be separated into fractions of differentiated abilities to provide available amino acids in the lower gut of ruminants. This knowledge is critical to develop feeding systems and to predict animal responses. We have measured during two growing seasons (1996 to 1997 and 1997 to 1998) the CP fractions of common vetch fresh forage with the objective being to assess the influence of maturity on concentration of CP fractions (as a percentage of total CP) and fraction yields. Fraction B2, which represents true protein of intermediate ruminal degradation rate, was the largest single fraction in common vetch forage (about 40% of CP across seasons and maturity stages). Soluble fractions (A plus B1) were less than 50% of total CP while the unavailable fraction C ranged from 4 to 8% of total CP. As a result, the remaining fraction B3 (true protein of very low degradation rate) only represented 2 to 9% of total CP. Concentration and yield of fraction B3 increased (P < 0.05) from flowering to pod-filling. Results showed that undegraded dietary protein represented a small proportion of total CP in common vetch forage. Moving the harvesting stage from flowering to the pod filling phase allowed for greater yield of undegraded dietary protein.     

Prediction of accuracy of estimated Mendelian sampling terms

This study describes a general framework for predicting the accuracy of Mendelian sampling terms when truncation selection is applied on best linear unbiased prediction (BLUP) estimated breeding values. A selection index approach is followed. The pseudo‐BLUP index is extended to include terms related to the Mendelian sampling term. Predicted accuracies are compared with those obtained through stochastic computer simulation. Good predictions for the accuracy of the Mendelian sampling term were obtained both at selection time and at convergence of long‐term contributions of selected candidates for a range of heritabilities and population structures. The prediction approach developed provides a key tool for obtaining predictions of genetic response from quadratic optimization that maximizes the rate of genetic progress while restricting the rate of inbreeding.     

Genetic and environmental parameters for racing time at different distances in Brazilian Thoroughbreds

The aim of the present study was to investigate genetic parameters for racing time in Thoroughbred horses racing at distances between 1000 and 1600 m subdivided into 100‐m intervals. The data provided by TURFETOTAL Ltda comprised races that occurred in the Gávea and Cidade Jardim race tracks over a period of 11 years (1992–2002) and consisted of 32 145 races and 238 890 time records. The variance components necessary to obtain the heritability and repeatability estimates of the traits studied were estimated with the MTDFREML program, and animal age at race (3 years old or younger, 4, 5 and older than 5 years), sex (male and female), number of races (1–32 145), and postposition at start (1–11) as fixed effects, and animal and permanent environmental random effects were included in a one‐trait animal model. Males were significantly superior to females at all distances. Excluding the 1100 m distance, animals 4 years of age were significantly faster than the mean of the other ages for all distances analysed. Horses older than 5 years showed a significantly lower performance than the mean of the other ages for all distances analysed, except for the 1100 m. Postpositions one and two did not differ significantly from one another for any of the distances analysed. These two inner positions both together varied from the other positions depending on race length. The components of additive genetic and permanent environmental variance varied in a similar way, tending to decrease with increasing racing distance, and the other temporary environmental variance almost doubled from 1000 to 1600 m. As was the case for the additive genetic and environmental variances, heritability and repeatability estimates tended to decrease with increasing distance, indicating that selection based on racing time will be less successful when the racing distance increases.