Modified live Edwardsiella ictaluri vaccine, AQUAVAC-ESC, lacks multidrug resistance plasmids

Plasmid-mediated antibiotic resistance was first discovered in Edwardsiella ictaluri in the early 1990s, and in 2007 an E. ictaluri isolate harboring an IncA/C plasmid was recovered from a moribund channel catfish Ictalurus punctatus infected with the bacterium. Due to the identification of multidrug resistance plasmids in aquaculture and their potential clinical importance, we sought to determine whether the modified live E. ictaluri vaccine strain in AQUAVAC-ESC harbors such plasmids, so that the use of this vaccine will not directly contribute to the pool of bacteria carrying plasmid-borne resistance. Antimicrobial sensitivity testing of the E. ictaluri parent isolate and vaccine strain demonstrated that both were sensitive to 15 of the 16 antimicrobials tested. Total DNA from each isolate was analyzed by polymerase chain reaction (PCR) using a set of 13 primer pairs specific for conserved regions of the IncA/C plasmid backbone, and no specific products were obtained. PCR-based replicon typing of the parent isolate and vaccine strain demonstrated the absence of the 18 commonly occurring plasmid incompatibility groups. These results demonstrate that the vaccine strain does not carry resistance to commonly used antimicrobials and provide strong support for the absence of IncA/C and other commonly occurring plasmid incompatibility groups. Therefore, its use should not directly contribute to the pool of bacteria carrying plasmid-borne resistance. This work highlights the importance of thoroughly investigating potential vaccine strains for the presence of plasmids or other transmissible elements that may encode resistance to antibiotics.     

Biofortification of tomato (Solanum lycopersicum) fruit with the anticancer compound methylselenocysteine using a selenocysteine methyltransferase from a selenium hyperaccumulator

Methylselenocysteine (MeSeCys) is an amino acid derivative that possesses potent anticancer activity in animals. Plants that can tolerate growth on soils with high Se content, known as Se hyperaccumulators, do so by converting inorganic Se to MeSeCys by the enzyme selenocysteine methyltransferase (SMT). A cDNA encoding the SMT from a Se hyperaccumulator was overexpressed in tomato (Solanum lycopersicum). Transgenic plants were provided with selenite or selenate to the roots during fruit development, and liquid chromatography-mass spectrometry was used to show that MeSeCys accumulated in the fruit but not in the leaves. Depending on the transgenic line and Se treatment, up to 16% of the total Se in the fruit was present as MeSeCys. MeSeCys was produced more effectively from selenite on a percentage conversion basis, but greater accumulation of MeSeCys could be achieved from selenate due to its better translocation from the roots. MeSeCys was heat stable and survived processing of the fruit to tomato juice.     

Specific serum antibody responses in channel catfish (Ictalurus punctatus) provide limited protection against Streptococcus ictaluri challenge

Passive immunization studies were conducted to determine the role of specific antibodies in immunity to Streptococcus ictaluri. Adult channel catfish (Ictalurus punctatus) were injected i.p. with tryptic soy broth as control or with 1.5 × 10(7)colony-forming units (cfu) S. ictaluri/fish at 0, 30, and 60 d, and serum was collected 90 d after the original challenge. Fish were passively immunized by i.p. injection with serum from the tryptic soy broth (TSB) control group, anti-S. ictaluri serum from fish immunized three times and sampled at 90 d (SSI), or heat-inactivated anti-S. ictaluri serum from fish immunized three times and sampled at 90 d (HISSI). These passively immunized fish were then challenged 72 h later with 1.5 × 10(8)cfu S. ictaluri/fish. Over 21 d, the mean cumulative percent survival was 43.3 (TSB), 63.3 (SSI), and 50.0 (HISSI). A significant difference in cumulative percent survival was noted between the TSB and the HISSI groups, and significant differences were noted between these groups and the SSI group. Serum obtained from immunized fish 72 h after passive immunization exhibited increased anti-S. ictaluri antibody levels. Twenty-one days after the challenge, the HISSI and SSI group antibody levels significantly increased above their corresponding pre-challenge levels. No significant (r(2)=0.0806; P<0.5985) correlation between increased pre-challenge specific serum antibody levels and survival after challenge was demonstrated when analyzing the control and passive immunization groups. The results indicate that both specific anti-S. ictaluri antibodies and non-specific immune responses are important for protection against S. ictaluri.     

Sub-cellular localisation of alkaline phosphatase activity in the cytoplasm of tammar wallaby (Macropus eugenii) neutrophils and eosinophils

Alkaline phosphatase (ALP) has been used in studies of neutrophil morphology and function as a marker for identifying different granule populations. In human neutrophils, ALP is found within secretory vesicles, a rapidly mobilisable vesicle population important for upregulating membrane receptors during early activation. Intra-cellular ALP activity in the heterophils of rabbits and guinea pigs, in contrast, is found only in secondary granules. The neutrophils and eosinophils of tammar wallabies (Macropus eugenii) have previously been reported to contain large amounts of ALP activity when stained using routine cytochemical techniques. To define the subcellular location of ALP in this species, cell suspensions were examined using cerium chloride cytochemistry and transmission electron microscopy (TEM). ALP was found in 2 distinct cytoplasmic compartments. One compartment displayed morphology consistent with a subpopulation of secondary granules while a second tubulo-vesicular population appeared similar to the secretory vesicles of human neutrophils. Thin tubular vesicles containing ALP were also identified within the cytoplasm of tammar wallaby eosinophils. Large numbers of ALP-containing vesicles have not been recognised previously in eosinophils and this may represent a novel cytoplasmic compartment. In both cell types, ALP-containing structures showed alteration in morphology following stimulation with N-formyl-Met-Leu-Phe (fMLP) and PMA.     

Nitrogen and phosphorus availability from liquid dairy manure to potatoes

The effective use of manure nutrients requires an accurate assessment of their availability to the specific crops being grown. A fertilizer equivalence approach was used to evaluate the availability of nitrogen (N) and phosphorus (P) from liquid dairy manure to potatoes in field experiments conducted in northeast Wisconsin in 2000, 2001, and 2002. Crop responses to moderate and high rates of liquid dairy manure (93,500 and 187,000 L ha^?1) were compared to results obtained from N or P fertilizer applied at five rates (0 to 269 kg ha^?1). Availability estimates were made using the fertilizer equivalence method based on tuber yield, harvested tuber N or P concentration and uptake, petiole nitrate or total P concentration, and soil nitrate or extractable P phosphate levels. Results showed an apparent availability of manurial N from 10% to 40%, with an overall average across all 3 years and all parameters of 28.6% for the lower rate and 24.6% for the high rate. These values are slightly less than average availability measured where corn has been used as the test crop. Although all parameters did not show responses to P fertilizer additions in these trials, where estimates were possible, P availability ranged from 20% to 90% with an overall average across all 3 years of 55.4% for the low and 50.5% for the high manure rates. Despite potato being more shallow-rooted and somewhat less efficient in N and P use compared to corn, it appears that only minor adjustments to manurial nutrient availability estimates are warranted when potatoes are grown.     

Effect of oral trazodone on the minimum alveolar concentration of isoflurane in dogs

Objective

To determine the effect of oral trazodone on the minimum alveolar concentration (MAC) of isoflurane in dogs.

Effect of Artificial Diets on Growth,Survival and Condition of Adult Cuttlefish, <Emphasis Type="Italic">Sepia officinalis</Emphasis> Linnaeus, 1758

The effects of artificial diets on growth and body condition of adult cuttlefish, Sepia officinalis were tested in two experiments. Supplemented prepared diets (fish myofibrillar protein concentrate) were fed during a 30-day and a 21-day experiments. Growth, feeding rate and food conversion of group-reared cuttlefish were analyzed. The first of these experiments tested four artificial diets, made with increasing levels of lysine, on adult cuttlefish. According to the chemical analysis, diets 1–3 had limiting concentrations of lysine and other essential amino acids (compared to mantle composition of the cuttlefish), while diet 4 was the only one where almost all essential amino acids were present in concentrations similar or higher than the ones present in cuttlefish mantle. A second experiment was conducted by isolating 16 adult cuttlefish individually, and feeding them the same four artificial diets, in order to obtain individual data. During Experiment 1, only the diet with the best chemical score (diet 4) produced growth (p < 0.05), with a mean instantaneous growth rate (MIGR) of 0.30% wet body weight (BW) d⁻¹. Similarly, individually reared cuttlefish fed diet 4 produced the highest IGR’s (0.26, 0.38 and 0.48% BW d⁻¹) and grew larger (p < 0.01). Comparison of cuttlefish fed the artificial diets vs. thawed shrimp and unfed cuttlefish indicated that cuttlefish fed the artificial diets were in an intermediate state. Growth rates obtained with the artificial diets (<0.4% BW d⁻¹) were considerably lower compared to natural prey, live or frozen, reported by other authors.     

Titres of precocene ii (6, 7-dimethoxy-2, 2- dimethylchromene) and its metabolites in the haemolymph of larvae of the african armyworm Spodoptera exempta (walker) following topical treatment with the compound

No precocious metamorphosis or other morphogenetic effects were seen following topical treatment of fifth instar Spodoptera exempta with pre- cocene II. [3H]-Precocene II penetrated the larvae rapidly following topical application. An uptake of 47% was recorded in the first hour and an average of 27% for the first 3 h. Radioactivity in the haemolymph reached a maximum 4 h after treatment at 6–8% of the applied dose. A maximum titre of precocene of 1.1 × 10^?4 _M was observed 2 h after treatment and a biological half-life of around 1 h was recorded for the first 6 h. The metabolite precocene 11–3, 4-dihydrodiol appeared in the haemolymph with a maximum titre of 1.1 × 10^?4 _M at 4 h suggesting metabolism via the 3, 4-epoxide. At the same time an unidentified metabolite was observed possibly corresponding to the 6- and 7-(O)-desmethylated products described by others. Very large quantities of highly polar materials were observed in the haemolymph throughout the experiment and it is thought that these corresponded to conjugates. The evidence suggests that peripheral detoxication mechanisms do not operate sufficiently rapidly to prevent the establishment of a high titre of precocene. This, however, did not affect the corpora allata in this insect, whereas in a sensitive species that has been studied by other workers, necrosis of the glands would have resulted. Alternative explanations for insensitivity are discussed.