Novel PCR primers for detection of genetically diverse virulent <Emphasis Type="Italic">Agrobacterium tumefaciens</Emphasis> biovar 1 strains

Novel PCR primers were developed to amplify a 243-bp fragment of an intergenic region between gene 5 and tms2 on the T-DNA of Agrobacterium tumefaciens biovar 1. These primers exhibit 100% positive correlation with strain virulence, 100% negative correlation with avirulence, and did not generate extraneous bands, thus facilitating robust real-time PCR detection.     

Lymphoid follicular cloacal inflammation associated with a novel herpesvirus in juvenile alligators (Alligator mississippiensis).

Multifocal hyperemic nodules and plaques associated with the cloacal mucosa of juvenile alligators (Alligator mississippiensis) at a public aquarium were investigated. Grossly, pale pink to dark red multifocal, circular lesions of varying degrees of severity were identified on the cloacal and, in males, phallus mucosa. Cloacal mucosa biopsies were obtained from 2 of the alligators. These samples were examined histologically and by polymerase chain reaction (PCR) using consensus primers targeting a conserved region of the herpesvirus polymerase gene. Microscopically, the lesions were characterized as submucosal lymphoid follicles with hyperemia and hemorrhage. No inclusion bodies were observed. Minimal to no anisokaryosis was present, and no etiologic agents were identified. Through PCR, a band consistent in size with herpesvirus was observed. Tissues showing similar clinical, histopathologic, and PCR findings were collected from animals at an alligator farm several months later. Sequencing of the PCR amplicon resulted in a 180-base pair sequence that shared 85% sequence identity with tortoise herpesvirus-1.     

Disseminated Intravascular Coagulation in Dogs: Review of the Literature

Diagnosis of Disseminated Intravascular coagulation (DIC) is controversial in both human and veterinary medicine. This article reviews the available literature in human and veterinary medicine regarding the etiology, pathophysiology, and diagnosis of DIC with emphasis on the diagnosis of DIC in dogs. Controversy surrounding the diagnosis of DIC arises from the complex nature of the disease itself, in addition to the absence of a consensus strategy for laboratory testing. The available literature indicates that dogs diagnosed with DIC possessed various hemostatic function testing abnormalities, typically possessed an underlying disease that predisposed to DIC, but may or may not have had clinically identifiable hemorrhage or thrombosis. Additionally, the hemostatic function testing utilized in diagnosis is not uniform. Generalizations about the usefulness of individual assays or diagnostic strategies cannot be formulated because of the marked diversity of the types of cases studied, as well as the small number of cases reported in the literature. (Vet Emerg & Crit Care, 1998; 8: 29–45)     

Magnesium: a quick reference.

This article serves as a quick reference for the distribution, handling, and supplementation of magnesium. It also lists the manifestations and causes of magnesium deficit and provides criteria for the diagnosis of a magnesium deficit.     

Evaluation of cGnRH IIa for induction spawning of two ornamental Epalzeorhynchos species

The administration of exogenous hormones for induction spawning of captive fish is a common practice throughout the finfish aquaculture industry. Induced spawning protocols for ornamental cyprinids commonly rely on sGnRH IIIa, commercially available as Ovaprim. In this study, the efficacy of an alternative spawning aid, cGnRH IIa, was evaluated relative to Ovaprim, in two commonly cultured ornamental Cyprinids: the Redtail Sharkminnow Epalzeorhynchos bicolor and the Rainbow Shark E. frenatum. Broodstock of each species were injected with either a positive control (0.5 µl/g Ovaprim), a negative control (propylene glycol) or one of three doses of cGnRH IIa (50, 100, or 200 µg/kg). Following spawning aid injection, ovulation success, fecundity, fertilization success, embryo diameter, hatch success and larval notochord length were evaluated. Ovulation success (82 ± 8%) was statistically similar to the positive control group for all experimental doses of cGnRH IIa in E. bicolor, while 50 and 100 µg/kg cGnRH IIa doses resulted in significantly higher ovulation success (100.0% and 83.0% respectively) than the positive control (17.0%) in E. frenatum. All other parameters did not vary significantly among treatments. These results indicate cGnRH IIa can be successfully used as a spawning aid in both species.     

Comparative renal anatomy of exotic species.

All living organisms consume nutrients that are required for the production of both tissue and energy. The waste products of this process include nitrogenous materials and inorganic salts. They are removed from the body by excretory organs, which in vertebrate shave developed into kidneys and into salt glands in some birds and reptiles. Many invertebrates use a series of excretory organs called nephridia to perform the same function. Even though they perform similar functions, there is no evolutionary connection between invertebrate nephridia and vertebrate kidneys. Both evolved independently.     

Segregation of axial motor and sensory pathways via heterotypic trans-axonal signaling

Execution of motor behaviors relies on circuitries effectively integrating immediate sensory feedback to efferent pathways controlling muscle activity. It remains unclear how, during neuromuscular circuit assembly, sensory and motor projections become incorporated into tightly coordinated, yet functionally separate pathways. We report that, within axial nerves, establishment of discrete afferent and efferent pathways depends on coordinate signaling between coextending sensory and motor projections. These heterotypic axon-axon interactions require motor axonal EphA3/EphA4 receptor tyrosine kinases activated by cognate sensory axonal ephrin-A ligands. Genetic elimination of trans-axonal ephrin-A --> EphA signaling in mice triggers drastic motor-sensory miswiring, culminating in functional efferents within proximal afferent pathways. Effective assembly of a key circuit underlying motor behaviors thus critically depends on trans-axonal signaling interactions resolving motor and sensory projections into discrete pathways.     

Seasonal flows on warm Martian slopes

Water probably flowed across ancient Mars, but whether it ever exists as a liquid on the surface today remains debatable. Recurring slope lineae (RSL) are narrow (0.5 to 5 meters), relatively dark markings on steep (25° to 40°) slopes; repeat images from the Mars Reconnaissance Orbiter High Resolution Imaging Science Experiment show them to appear and incrementally grow during warm seasons and fade in cold seasons. They extend downslope from bedrock outcrops, often associated with small channels, and hundreds of them form in some rare locations. RSL appear and lengthen in the late southern spring and summer from 48°S to 32°S latitudes favoring equator-facing slopes, which are times and places with peak surface temperatures from ~250 to 300 kelvin. Liquid brines near the surface might explain this activity, but the exact mechanism and source of water are not understood.     

Genetic analysis of streptomycin-resistant (Sm(R)) strains of Erwinia amylovora suggests that dissemination of two genotypes is responsible for the current distribution of Sm(R) E. amylovora in Michigan

Streptomycin-resistant (Sm(R)) strains of the fire blight pathogen Erwinia amylovora were first isolated in southwest Michigan in 1991. Since that time, resistant strains have progressed northward to other apple-producing regions in the state. A total of 98.7% of Sm(R) strains isolated between 2003 and 2009 in Michigan harbored the strA-strB genes on transposon Tn5393. strA and strB encode phosphotransferase enzymes that modify streptomycin to a nonbactericidal form. Mutational resistance to streptomycin, caused by a point mutation-mediated target-site alteration of the ribosomal S12 protein, occurred in 1.3% of E. amylovora strains from Michigan. Tn5393 was originally introduced to E. amylovora on the plasmid pEa34; thus, the first Sm(R) strains isolated contained both pEa34 and the ubiquitous nonconjugative plasmid pEA29. More recently, we have observed Sm(R) strains in which Tn5393 is present on pEA29, suggesting that the transposon has moved via transposition from pEa34 to pEA29. Almost all of the strains containing Tn5393 on pEA29 had lost pEa34. Of 210 pEA29::Tn5393 plasmids examined, the transposon was inserted at either nucleotide position 1,515 or 17,527. Both of these positions were in noncoding regions of pEA29. Comparative sequencing of the housekeeping genes groEL and potentially variable sequences on pEA29 was done in an attempt to genetically distinguish Sm(R) strains from streptomycin-sensitive (Sm(S)) strains isolated in Michigan. Only 1 nucleotide difference within the total 2,660 bp sequenced from each strain was observed in 2 of 29 strains; multiple sequence differences were observed between the Michigan strains and E. amylovora control strains isolated in the western United States or from Rubus spp. Alterations in virulence observable using an immature pear fruit assay were detected in three of eight Sm(R) strains examined. Our current genetic data indicate that only two Sm(R) strain genotypes (strains containing pEA29::Tn5393 with Tn5393 inserted at either nucleotide position 1,515 or 17,527 on the plasmid) are responsible for the dissemination of Tn5393-encoded streptomycin resistance in Michigan, and that the Sm(R) and Sm(S) strains in Michigan compose a homogenous group.     

Decomposition and nitrogen release of understorey plant residues in biological and integrated apple orchards under field conditions in New Zealand

In grassed-down apple orchards in New Zealand, the understorey vegetation is usually mown and the plant residues are returned to the orchard floors as a source of nutrients. It is, therefore, important to determine the decomposition pattern and the rate of N release from understorey plant residues. In this study, the decomposition and N release of surface-placed understorey plant residues were determined in the field and compared across treatments of grassed-down biological (BFP) and integrated fruit production (IFP) orchards in two different locations (Lincoln and Clyde) in New Zealand using the litterbag technique. At Lincoln, the field experiment was a randomised complete block design with three different treatments (two BFPs, one IFP) each with three replicates; while at Clyde, the field experiment consisted of non-replicated apple orchard plots with three treatments (two IFPs, one BFP). A comparison was also made between surface-placed and soil-buried understorey plant residues in a BFP orchard at one location. Samples of understorey plant residues collected from orchard mowings in the respective treatments were returned to the same treatment plot in litterbags and retrieved at intervals of 90 days for 360-450 days. Results showed that the single exponential decay model, Y=A₀ e^-kt, accounted significantly (PА.001) for 97-99% of the variation in the decomposition and N release patterns, which ranged from 6.0᎒^-3-9.6᎒^-3 day^-1 and 7.0᎒^-3-13.0᎒^-3 day^-1, respectively. Half-lives for C and N of residues were approximately 70-120 days and 50-110 days, respectively. Soil-buried plant residues showed more rapid decomposition and N release compared with those of surface-placed plant residues (80% vs. 54% in 90 days). In general, plant residue decomposition and N release were significantly more rapid in IFP than in BFP treatments (13.0᎒^-3 vs 7.0᎒^-3 day^-1 for N release). Overall, differences in plant residue decomposition and N release rates related to understorey plant residue quality and treeline management practices rather than the orchard system as a whole.