Quantitative loop-mediated isothermal amplification method for the detection of <Emphasis Type="Italic">Vibrio nigripulchritudo</Emphasis> in shrimp

Vibrio nigripulchritudo is considered one of the major pathogens threatening shrimp aquaculture. In this study, we developed a novel and highly specific quantitative loop-mediated isothermal amplification (Q-LAMP) assay. A set of four specific primers were designed targeting the V. nigripulchritudo intergenic spacer region. The reaction time and temperature were optimized for 60 min at 63°C. Quantitative analysis was then performed by measuring the turbidity of the reaction solution using a real-time turbidimeter, allowing for quantification of the initial DNA concentration with a sensitivity of 10² copy numbers equivalent to 2.3 colony forming units/ml or 0.3 fg/μl. The LAMP assay was able to specifically detect two representative strains of V. nigripulchritudo, whereas other Vibrio and non-Vibrio species were not amplified. A standard curve was generated for V. nigripulchritudo by plotting the threshold time (T _t) versus the log of bacterial number. A high correlation coefficient (R ² = 0.9749) was observed for the Q-LAMP reaction. In conclusion, Q-LAMP assay is a sensitive, rapid, and simple tool that can be used for the detection and quantification of V. nigripulchritudo in shrimp, thereby facilitating surveillance of vibriosis infection.     

Is it a hindrance for an invasive aquatic species to spread across scattered habitat patches?

1. Despite their importance for aquatic biodiversity, ponds are among the most vulnerable freshwater habitats. Owing to their isolation in terrestrial environments, ponds are expected to be relatively well protected from biological invasions, but this depends on many factors. 2. The purpose of this study was to examine the influence of a range of variables (water quality, habitat, and landscape attributes) on the colonization of discrete ponds by a widespread aquatic invader, the red swamp crayfish Procambarus clarkii, which can disperse overland. 3. Investigations were conducted in two networks of ponds, each located in close proximity to a large invaded marsh. The two marshes under study differed in the length of time since the crayfish were introduced. 4. The proportions of colonized ponds and crayfish abundances were moderate in both networks, but higher in the network that had been invaded first. In both networks the distance to the marsh was the main predictor of pond colonization, considering similar energy costs to cross aquatic and terrestrial habitats for the recently invaded network, but assuming that dispersal was 25 times costlier across the terrestrial matrix than via streams for the earlier invaded network. Pond characteristics had no influence on crayfish occurrence in either network. Furthermore, predictions of pond invasion were lower for the recently invaded network. 5. The importance of the distance to the marsh indicates that natural dispersal was the main process of pond colonization by crayfish. Findings also suggested that overland dispersal was rare and costly. By contrast, streams were probably significant in facilitating crayfish dispersal. Differences between the two networks might arise from an invasion process still in progress in the recently invaded network. 6. From a management viewpoint, local actions are encouraged to prevent the spread of crayfish via streams. In addition, broader‐scale actions to mitigate other human disturbances would improve the outlook for pond biodiversity.     

Effect of an inactivated bluetongue serotype 8 vaccine on semen quality in rams

The aim of this study was to determine whether a single dose of an inactivated bluetongue virus serotype 8 (BTV-8) vaccine altered semen quality in rams. Twenty sexually mature rams were assigned to three experimental groups: two groups of four animals were vaccinated and a third group of four animals was unvaccinated. The first group included rams with a history of natural BTV-8 infection in 2007 and the second and third groups included BTV-8 na?ve rams. Semen was collected prior to vaccination and for 4months post-vaccination. There were no significant differences in semen quality traits, including motility and concentration of spermatozoa, and percentages of living, normal dead and abnormal dead spermatozoa, between vaccinated and unvaccinated groups, or over time (P>0.05). The BTV-8 vaccine tested in this study did not appear to have any adverse effect on semen quality in rams.     

Isolation of Arcanobacterium hippocoleae from a case of placentitis and stillbirth in a mare

Based on the scarcity of reports in veterinary medicine literature, it appears that Arcanobacterium spp. are rarely isolated from horses. Recently, a single isolate from vaginal discharge in a mare was characterized as a new species, Arcanobacterium hippocoleae. The present report describes a case of necrosuppurative placentitis and stillbirth in an American Quarterhorse mare. Numerous colonies of irregular, Gram-positive, rod-shaped bacteria were observed by histological examination within fibrin lattice associated with placental lesions. Arcanobacterium hippocoleae was isolated in large numbers from the placenta, lung, and stomach contents. To the authors' knowledge, this is the first report of placentitis associated with Arcanobacterium spp. in a horse and the third reported isolation of A. hippocoleae from a horse.     

Juvenile mortality in captive lesser kudu (Tragelaphus imberbis) at Basle Zoo and its relation to nutrition and husbandry.

Since 1956, when the Basle Zoo (Switzerland) initiated the breeding of lesser kudu (Tragelaphus imberbis), 43% of the lesser kudu juveniles died before reaching an age of 6 mo. In this study, the objective was to obtain the pathological findings, nutritional history, and family tree information in order to evaluate the influence of husbandry on juvenile mortality in these animals. The main cause of death was white muscle disease (WMD), diagnosed in 14 cases (26%) of the deceased juveniles. Although enclosure size had remained constant and animal accessibility to the public was constantly high, both herd size and juvenile mortality had increased from 1956-2004. The diet consumed by the whole group in 2004 had deficient levels of vitamin E and selenium. The increasing linear trend of the mortality rate since the 1960s was significant, and there was a significant correlation between herd size and overall juvenile mortality. In contrast, there was no correlation between herd size and the occurrence of juvenile mortality associated specifically with WMD. Other investigated factors (sex, inbreeding, and season) had no significant effect on overall mortality up to 6 mo of age or on mortality associated with WMD. These results characterize both a dietary and a husbandry problem, and are supported by a lack of similar juvenile mortality in another facility where the diet was supplemented with vitamin E, animal numbers were kept low, and the enclosure structure offered more retreat options for the animals.     

Non‐spherical pearl layers in the Polynesian ‘black‐lipped’ Pinctada margaritifera: The non‐nacreous deposits compared to microstructure of the shell growing edge

Cultivated pearls frequently exhibit morphological irregularities making obvious that mineral deposition was irregularly distributed onto nucleus surface. Taking advantage of experimental cultivations with short durations (from 10 days to few months), these irregular deposits predating occurrence of the nacre were investigated in Polynesian pearls by biochemical characterizations and a series of physical methods. Diversity in the resulting data suggests that various in‐depth alterations of the biomineralization mechanism may have occurred during the grafting process, leading to diversity in the biochemical pathways to nacreous deposition. This allows a precise discussion of current views about pearl formation. The “reversed shell theory” is formally disproved through point to point comparison with development of the shell growing edge. Similarity of pearl formation with “regeneration” or “shell repair” is also discussed, emphasizing the differences between these concepts.     

Effect of oral administration of low doses of pentobarbital on the induction of cytochrome P450 isoforms and cytochrome P450-mediated reactions in immature Beagles

OBJECTIVE: To determine the effect of oral administration of low doses of pentobarbital on cytochrome P450 (CYP) isoforms and CYP-mediated reactions in immature Beagles. ANIMALS: 42 immature (12-week-old) Beagles. PROCEDURE: Dogs were grouped and treated orally as follows for 8 weeks: low-dose pentobarbital (50 microg/d; 4 males, 4 females), mid-dose pentobarbital (150 microg/d; 4 males, 4 females), high-dose pentobarbital (500 microg/d; 4 males, 4 females), positive-pentobarbital control (10 mg/kg/d; 2 males, 2 females), positive-phenobarbital control (10 mg/kg/d; 2 males, 2 females), and negative control (saline 10.9% NaCl] solution; 5 males, 5 females). Serum biochemical and hematologic values were monitored. On necropsy examination, organ weights were determined, and histologic evaluation of tissue sections of liver, kidney, small intestine, testes, epididymis, and ovaries was performed. Hepatic and intestinal drug-metabolizing enzyme activities were measured, and relative amounts of CYP isoforms were determined by western blot analysis. RESULTS: The amount of a hepatic CYP2A-related isoform in dogs from the high-dose pentobarbital treatment group was twice that of dogs from the negative control group. CYP2C was not detectable in small intestinal mucosa of dogs from the negative control group; measurable amounts of CYP2C were found in dogs from the various (low-, mid-, and high-dose) pentobarbital treatment groups and from positive-pentobarbital and positive phenobarbital control groups. Several CYP-mediated reactions increased in a dose-dependent manner. The lowest calculated effective dose of pentobarbital ranged from 200 to 450 microg/d. CONCLUSIONS AND CLINICAL RELEVANCE: Several CYP isoforms and their associated reactions were induced in dogs by oral administration of low amounts of pentobarbital.     

Pasting and Thermal Properties of Flours from Oat Lines with High and Typical Amounts of β‐Glucan

Experimental oat lines high in β‐glucan (6–7.8%) and traditional lines (3.9–5.7% β‐glucan) were used to evaluate the effect of β‐glucan on pasting (by rapid viscoanalysis) and thermal properties (by differential scanning calorimetry) of oat flours. Significant correlations established between β‐glucan concentration and the pasting parameters after amylolysis demonstrated the role of β‐glucans in oat pasting. The relative decrease of peak viscosity (PV) observed after enzymatic removal of β‐glucans was correlated with β‐glucan concentration (r = 0.880, P < 0.010) and reconfirmed their contribution to pasting. A significant increase of PV with β‐glucan concentration obtained under conditions of either autolysis (deionized water used for dispersion) (r = 0.89, P < 0.010) or inhibition (silver nitrate solution used for dispersion) (r = 0.91, P < 0.001) might be explained by an increase in water retention capacity caused by the β‐glucans. Predictive models of β‐glucan concentration based on the whole pasting profile or selected profile regions were developed using partial least squares (PLS) regression.Prediction of β‐glucan based on the whole profile obtained in the silver nitrate solution was the most effective (r = 0.93, correlation coefficient of predicted vs. analyzed β‐glucans, P < 0.050). No correlations were observed between the thermal properties of oat flours and the β‐glucan concentration.