Sialidase activity in Mycoplasma synoviae

Eleven strains of the avian pathogen Mycoplasma synoviae were evaluated for the presence of sialidase activity with the use of the fluorogenic substrate 2'-(4-methylumbelliferyl)-alpha-D-N-acetylneuraminic acid and the sialidase inhibitor 2-deoxy-2,3- didehydro-N-acetylneuraminic acid. The kinetics of in vitro growth in modified Frey medium were also assessed for each strain. Five strains had been isolated from clinically symptomatic chickens, and strains WVU 1853T and K3344 have been demonstrated to be capable of reproducing disease in specific-pathogen-free chickens. All strains exhibited sialidase activity, although the amount varied 65-fold among strains (P < 0.0001) from 1.3 x 10(-7) to 2.0 x 10(-9) activity units per colony-forming unit. Strains originally isolated from clinically symptomatic birds had more (P < 0.05) sialidase activity than strains from asymptomatic birds. Strain WVU1853T exhibited the most sialidase activity (P < 0.0001) and grew to the highest culture density (P < 0.0001) among strains, but across strains, the rank correlation of growth rate with sialidase activity was not significant. Negligible activity was detected in conditioned culture supernatant fluid. This is the first report of sialidase activity in pathogenic strains of M. synoviae, which suggests a potential enzymatic basis for virulence of the organism.     

Variation in E(rns) viral glycoprotein associated with failure of immunohistochemistry and commercial antigen capture ELISA to detect a field strain of bovine viral diarrhea virus

Bovine viral diarrhea virus (BVDV) affects cattle populations causing clinical signs that range from subclinical immunosuppression to severe reproductive and respiratory problems. Detection and removal of persistently infected (PI) calves is the single most important factor for control and eradication of BVDV. Current testing strategies to detect PI calves rely heavily on immunohistochemistry (IHC) and a commercially available antigen capture ELISA (ACE) assay. These viral assays depend on 1 or 2 monoclonal antibodies which target the E(rns) glycoprotein of BVDV. The sensitivity and specificity of these two tests have been reported previously. The purpose of this research was to characterize a strain of BVDV (AU501) that was undetectable using IHC and ACE based on a single monoclonal antibody, but was consistently detected in samples from a Holstein steer using virus isolation and PCR testing. Sequencing of this AU501 viral isolate revealed a unique mutation in the portion of the genome coding for the E(rns) glycoprotein. This unique field strain of BVDV demonstrates the risk of relying on a single monoclonal antibody for detection of BVDV. Multiple testing strategies, including polyclonal or pooled monoclonal antibodies that detect more than one viral glycoprotein may be necessary to detect all PI calves and facilitate eradication of BVDV.     

Remote sensing of biotic and abiotic stress for irrigation management of cotton

The applicability of commercially available remote sensing instrumentation was evaluated for site-specific management of abiotic and biotic stress on cotton (Gossypium hirsutum L.) grown under a center pivot low energy precision application (LEPA) irrigation system. This study was conducted in a field where three irrigation regimes (100%, 75%, and 50% ET_c) were imposed on areas of Phymatotrichum (root rot) with the specific objectives to (1) examine commercial remote sensing instrumentation for locating areas showing biotic and abiotic stress symptomology in a cotton field, (2) compare data obtained from commercial aerial infrared photography to that collected by infrared transducers (IRTs) mounted on a center pivot, (3) evaluate canopy temperature changes between irrigation regimes and their relationship to lint yield with IRTs and/or IR photography, and (4) explore the use of deficit irrigation and the use of crop coefficients for irrigation scheduling. Pivot-mounted IRTs and an IR camera were able to differentiate water stress among irrigation regimes. The IR camera distinguished between biotic (root rot) and abiotic (drought) stress with the assistance of groundtruthing. The 50% ET_c regime had significantly higher canopy temperatures than the other two regimes, which was reflected in significantly lower lint yields when compared to the 75% and 100% ET_c regimes. Deficit irrigation down to 75% ET_c had no impact on lint yield, indicating that water savings were possible without reducing yield.     

Influence of water potential on nitrification and structure of nitrifying bacterial communities in semiarid soils

The objective of this study was to examine the relationship between soil water potential, nitrifier community structure and nitrification activity in semiarid soils. Soils were collected after a 5-month dry period (end of summer) and subsequently rewetted to specific water potentials and incubated for 7 days prior to analysis of nitrification activity and nitrifier community structure. The approach used in this study targeted a 491bp segment of the amoA gene which encodes the active site of the ammonia monooxygenase enzyme, which is the key enzyme for all aerobic ammonia oxidisers. amoA serves as a useful target for environmental studies since it is both specific and universal for all ammonia oxidisers and reflects the phylogeny of the ammonia oxidisers. Our results suggest that in semiarid soils water potential plays a key role in determining the structure of ammonia oxidising bacteria (AOB), and that additionally AOB community structure is correlated to potential nitrification rate in these soils.     

Micro-scaled high-throughput digestion of plant tissue samples for multi-elemental analysis

Background  

Quantitative multi-elemental analysis by inductively coupled plasma (ICP) spectrometry depends on a complete digestion of solid samples. However, fast and thorough sample digestion is a challenging analytical task which constitutes a bottleneck in modern multi-elemental analysis. Additional obstacles may be that sample quantities are limited and elemental concentrations low. In such cases, digestion in small volumes with minimum dilution and contamination is required in order to obtain high accuracy data.