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.     

Pharmacokinetics of enrofloxacin in neonatal kittens

OBJECTIVE: To determine the pharmacokinetics of enrofloxacin in neonatal kittens and compare the pharmacokinetics of enrofloxacin in young and adult cats. ANIMALS: 7 adult cats and 111 kittens (2 to 8 weeks old). PROCEDURE: A single dose of 5 mg of enrofloxacin/kg was administered to adults (i.v.) and kittens (i.v., s.c., or p.o.). Plasma concentrations of enrofloxacin and its active metabolite, ciprofloxacin, were determined. RESULTS: The half-life of enrofloxacin administered i.v. in 2-, 6-, and 8-week-old kittens was significantly shorter and its elimination rate significantly greater than that detected in adults. The apparent volumes of distribution were lower at 2 to 4 weeks and greater at 6 to 8 weeks. This resulted in lower peak plasma concentration (Cmax) at 6 to 8 weeks; however, initial plasma concentration was within the therapeutic range after i.v. administration at all ages. Compared with i.v. administration, s.c. injection of enrofloxacin in 2-week-old kittens resulted in similar Cmax, half-life, clearance, and area under the curve values. Enrofloxacin administered via s.c. injection was well absorbed in 6- and 8-week-old kittens, but greater clearance and apparent volume of distribution resulted in lower plasma concentrations. Oral administration of enrofloxacin resulted in poor bioavailability. CONCLUSIONS AND CLINICAL RELEVANCE: In neonatal kittens, i.v. and s.c. administration of enrofloxacin provided an effective route of administration. Oral administration of enrofloxacin in kittens did not result in therapeutic drug concentrations. Doses may need to be increased to achieve therapeutic drug concentrations in 6- to 8-week-old kittens.     

A behavioural approach of dominance establishment in tank‐held sea bream (Sparus aurata L.) under different feeding conditions

Dominance is a strong behaviour exhibited by farmed species that very often impinges on fish growth and welfare. This study presents a behavioural approach of dominance, where colour pattern differentiation was tested as a signal of dominance presence in small sea bream population. The phenotype of dominance was first described in detail, referring to vertical dark stripes and splayed fins. Fish were kept in triplicate tanks for 21 days, during which they were exposed to three feeding conditions (well fed, limited fed and no‐fed) and continuous video recordings. Each tank was stocked with 15 individuals (~30.34 ± 1.70 g), and they were confined to the half volume of the tank via a removable net pen. Percentage of dominants per population was found up to 40%, while duration was calculated to 53.52 ± 7.44 s. Dominance behaviour was further quantified via colour pattern differentiation on sea bream body (CDA, contrast of dominance appearance). The results demonstrated that the body colour of sea bream is directly linked to species social hierarchies and such variations are visual signal of dominance rank inside the population. The fish feeding conditions had an influence on the dominance presence, but not on the dominance rank (as measured by the CDA) between fish groups. The described study provides state‐of‐the‐art knowledge on sea bream dominance. In relevance to aquaculture, dominance quantification would be a reliable tool to evaluate non‐equal food distribution in tank‐held populations, thus, avoid non‐regulate growth performance of fish before transfer to sea‐cage installations.     

Hormonal changes during meiotic maturation and ovulation in the brook trout (Salvelinus fontinalis)

The plasma levels of estradiol-17 (E2), 17, 20-dihydroxy-4-pregnen-3-one (17,20-P) and gonadotropin (GTH) were measured in brook trout (Salvelinus fontinalis) during the period from the end of vitellogenesis to postovulation. Blood samples were taken according to specific stages of maturation, including germinal vesicle breakdown (GVBD) and ovulation. E2 levels were quite high (45 ng/ml) at the end of vitellogenesis (and prior to GVBD) and dropped precipitously by GVBD (2 ng/ml). They remained low through ovulation and postovulation. 17,20-P levels were low prior to GVBD (0.7 ng/ml) and increased dramatically at GVBD (148 ng/ml). The levels of 17,20-P remained high at ovulation (142 ng/ml) and then dropped significantly within 24 h to approximately half of the ovulatory values. They decreased even further by 7 days postovulation. GTH levels rose gradually through GVBD and ovulation from a postvitellogenic level of approximately 3 ng/ml to a 7 day postovulatory value of approximately 10 ng/ml. The overall results; 1) decrease in estradiol prior to GVBD, 2) increase in 17,20-P at GVBD and 3) gradual GTH rise through GVBD and ovulation, are similar to those reported for other salmonids.     

Leptin and leptin receptors during malaria infection in mice

Leptin, which is involved in a range of physiological processes, could be an important factor in the pathogenesis of malaria. We found that levels of leptin in serum and urine in Plasmodium berghei-infected mice increased progressively after infection, reaching a maximum value on day 6 post-infection. Serum values were approximately five-fold higher in infected mice than in non-infected controls. A similar relation was found for values of leptin in urine. Soluble leptin receptor levels also increased significantly in serum, more or less in line with the leptin increase. Our work represents the first report of visibly augmented leptin and soluble leptin receptor secretion in malarial infection.     

Influence of acclimation temperature on the osmotic regulation and survival of rainbow trout (Salmo gairdneri) rapidly transferred from fresh water to sea water

Certain physiological responses of two groups of rainbow trout acclimated at 10 and 17°C to a sudden change in salinity were investigated in order to determine the most appropriate temperature for direct transfer in salt water. Plasma osmolarities of fish acclimated at 17°C increased at a much faster rate following transfer and approached a much higher value than that reached by the fish acclimated at 10°C. The same was apparent for body tissue dehydration. Haematocrit values, the number of red blood cells, as well as red cell volume showed no significant difference between the two groups of fish for most of the experimental period. The larger increases in plasma osmotic pressure and tissue dehydration observed at 17°C are discussed in connection with the physiological adjustments accompanying higher temperature acclimation.     

Use of monthly collected milk yields for the detection of the emergence of the 2007 French BTV epizootic

Two culicoides-borne diseases, Bluetongue (BTV) and Schmallenberg, have emerged in the European cattle population since 2006. Other diseases transmitted by these vectors could emerge. This justifies the development of syndromic surveillance programs whereby one or several indicators would be routinely monitored for the early detection of emerging diseases. The aim of this study was to evaluate milk yield from milk recording in dairy cattle as an indicator to be included in an emerging disease surveillance system. It was hypothesized that emergences would result in episodes of low milk production clustered in space and time. The 2007 BTV epizootic in France was used as a case study. Because it had already emerged in neighbouring countries, the disease emergence was expected and notification was mandatory. Herd-test-day milk productions were predicted for the entire country for 2006 and 2007 from herd historical data using linear mixed models. The differences between observed and predicted milk productions were averaged per week and per municipality and used as input for a space-time prospective scan statistic. Log likelihood ratios (LLR) associated with clusters were used to define alarms. The threshold chosen was a trade-off between detection timeliness and the number of false alarms per week. The first four BTV notifications occurred on the 12th (two notifications), 13th and 27th of July 2007. The 12th of July was considered to be the date of emergence. Alarms occurring before the 1st of March 2007 were considered to be false alarms. Using an LLR of 50, there were an average of 1.7 false alarms per week and the BTV emergence was detected seven weeks after emergence. Using an LLR of 100, there were an average of 0.8 false alarms per week and the BTV emergence was detected 9 weeks after emergence. Detection may have been delayed because of a discontinuation of milk recording between mid-July and mid-August. The first cluster with an LLR > 100 located in the emergence area was further investigated. A difference between observed and predicted production of >1 kg/cow/day was observed around the time of emergence. However, a difference of equal magnitude was observed during the year preceding the outbreak. Milk production predicted from herd history alone did not allow the detection of the 2007 BTV emergence in France. Further research should be conducted on improving the prediction of test-day milk yield and on combining it with other indicators based on routinely collected data.     

Pharmacokinetics of oxolinic acid in gilthead sea bream, Sparus aurata L.

This is the first study on the pharmacokinetic parameters of oxolinic acid (OA) in gilthead sea bream, Sparus aurata L. The kinetic profile of OA was studied after a single intravascular injection (20 mg kg⁻¹) in 100 g fish at 20 °C. The distribution half-life ( t _1/2α) and the elimination half-life ( t _1/2β) of the drug were found to be short (0.51 and 12.60 h, respectively). The drug penetration from the plasma to the tissues was adequate as the apparent volume of distribution of the drug at steady-state ( V _d(ss)) was found to be 2.11 L kg⁻¹. The mean residence time ( MRT ) of OA was short (14.25 h) and the total clearance rate ( Cl _T) of the drug was low (0.15 L kg⁻¹ h⁻¹). The bioavailability ( F %) of OA following oral administration (30 mg kg⁻¹) was also low (14%). Maximum values were observed for muscle at 0.5 h after injection, with levels declining as with subsequent sampling. At the first two time points (0.5 and 1 h) plasma levels of OA were higher than muscle, however, the reverse was evident for subsequent samples. Following oral administration, highest muscle levels were found at 16 h and, with the exception of the 24-h sampling, muscle OA concentrations were higher than plasma at all time points. The fast elimination of OA suggests short withdrawal times with reference to human consumption of treated fish.     

Flumequine depletion from muscle plus skin tissue of gilthead seabream (Sparus aurata L.) fed flumequine medicated feed in seawater at 18 and 24 °C

We examined flumequine depletion from muscle plus skin of gilthead seabream held in seawater at 18 and 24 °C. Seven groups of 10 fish each were sampled at intervals ranging from 24 to 168 h after in-feed administration of flumequine at 35 mg/kg/day for 5 days. Muscle plus skin tissue samples were analyzed for flumequine by high-performance liquid chromatography and fluorescence detection (HPLC-SFD). Parent flumequine concentrations declined rapidly from muscle plus skin after dosing with elimination half-lives of t_1/2=22.14 and 21.43 h at 18 and 24 °C, respectively. Withdrawal periods for the maximum residue limit (MRL) of 600 μg/kg flumequine in muscle plus skin at 95% tolerance limit were 106.08 and 75.84 h at 18 and 24 °C, respectively, after treatment.     

Effects of early re-spacing on the physical and mechanical properties of Sitka spruce structural timber

The effects of early re-spacing on the physical and mechanical properties of Sitka spruce (Picea sitchensis [Bong.] Carr.) structural-dimensioned timber were studied using material from a fully replicated 57-year-old trial located in Northern Ireland, which had been thinned at age 11 years. Basic density, distortion (spring, twist and bow), modulus of elasticity (MOE) and modulus of rupture (MOR) in bending of structural timber from four different re-spacing treatments (1.83 m × 3.66 m, 3.66 m × 3.66 m, 3.66 m × 5.49 m and 5.49 m × 5.49 m) were compared with those of timber from a control (1.83 m × 1.83 m). Re-spacing intensity had a significant effect on both modulus of elasticity (p = 0.006) and modulus of rupture (p = 0.009), but not on basic density (p = 0.379) or distortion (p > 0.200). Timber from the two widest re-spacing treatments failed to meet the requirements for the C14 strength class, while timber from the control met the requirements for the C16 strength class. Both MOE and MOR were significantly and negatively associated with knot size and frequency, which in turn increased with re-spacing intensity. Overall, re-spacing intensity only explained approximately 10% of the total amount of variation in both MOE and MOR, with most of the variation due to inter- and intra-tree differences within a treatment. Based on these results, re-spacing of Sitka spruce to a residual stand density of less than 900 trees/ha (3.33 m × 3.33 m) is not recommended if the goal of management is to produce C16 structural timber.