Chitinase production by an Arthrobacter sp. lysing cells of Fusarium roseum

An Arthrobacter sp. which actively lysed Fusarium roseum was found to liberate chitinase (E.C. 3.2-1.14, chitin glycanohydrolase), an enzyme essential for the hydrolysis of chitin, a major component of fusarial hyphal walls. Factors involved in the production of chitinase were investigated by modifying culture conditions and assaying for enzyme activity. Production occurred on colloidal chitin as well as on native chitin supplemented with yeast extract or peptone. Enzyme production paralleled growth; liberation of enzyme took place during the log phase with the maximum yield being obtained at the stationary phase. Addition of the non-ionic surfactant, polyoxyethylene sorbitan monooleate (Tween 80) increased enzyme yield. An inverse relationship was found between the amount of enzyme produced and the quantity of n-acetyl-glucosamine liberated. The enzyme was generally not produced when grown on various other carbohydrates. These findings suggest that chitinase is inducible and that chitin breakdown is regulated by a repressor-inducer mechanism.Initial hydrolysis rates of colloidal chitin were proportional to the concentration of chitinase used. Optimal pH and temperature for enzyme activity were 4.9 and 50°C, respectively. Purification of the chitinase was obtained by (NH₄)₂SO₄ precipitation followed by DEAE-cellulose and Sephadex chromatography, achieving a 12-fold increase in specific activity.     

Horizontal transmission of Mycobacterium avium subsp. paratuberculosis in cattle in an experimental setting: calves can transmit the infection to other calves

In September 2001, two subsequent transmission experiments both lasting 3 months were carried out to study cow-calf transmission of Mycobacterium avium subsp. paratuberculosis (Map) (Period 1), followed by calf-calf transmission of the infection (Period 2). Every 2 weeks, serum, heparinised blood and faecal samples were collected from all animals. After these experiments, the 20 calves were housed individually for more than 3 years to be able to detect the infection status and excretion pattern of each animal. In autumn 2004, the animals were inseminated, to observe a possible increase in faecal excretion of Map shortly before expected calving. One month before the expected calving date in 2005, animals were slaughtered and several tissues per cow and unborn calf were sampled for culture. The results indicate that horizontal cow-calf transmission is readily achieved (Period 1). At the highest infection pressure (six shedding cows of which three high shedders in Period 1) all five calves excreted Map in their faeces during Period 1 (shortly after infection), and four of these calves during Period 2 (when the shedding cows were absent). After that, excretion became less frequently. Horizontal calf-calf transmission did take place (Period 2), as the four donor-calves infected two receiver-calves. Transmission rates during the 3 months periods were quantified as a reproduction ratio R. The R [95% CI] of cow-calf and calf-calf transmission were estimated as 2.7 [1.1, 6.6] and 0.9 [0.1, 3.2] new infections per infectious animal during 3 months.     

Remediation of Soils Contaminated with Molybdenum using Soil Amendments and Phytoremediation

For ruminants, there is a narrow span betweennutritional deficiency of Molybdenum and its potential toxicity. Molybdenosis occurs among cattlefeeding on forage with Mo concentrations above10 μg g^-1 or a Cu:Mo ratio <2. In the areaunder investigation forage Mo contents in the valleyare as high as 180 μg g^-1 due to industrialpollution, while the alpine pastures, where cattlegraze during summer, are nutrient (e.g. Cu) deficient.When driven to the valley pastures in fall, animalsoften fell ill with molybdenosis, and several died.The aim of the study was to remediate this 300 ha area. Two approaches were attempted: (1) to reduce theMo contents of the forage by immobilizing soil Mo, and(2) to increase plant Mo contents by mobilizing soilMo to increase plant Mo which may then be removed fromthe system (phytoremediation).In a soil extraction experiment we demonstrated thatphosphate fertilization, ammonium sulfate amendment,vermiculite, humic acid and sewage sludge increased Moleaching by 30 to 110%. Fe-humate and Mn-humateapplication decreased Mo in the leachate from96 μg L^-1 to 55 and 7 μg L^-1,respectively. Plant Mo contents were increased up to170% by P-fertilizer and up to 150% by vermiculite.Conversely, sewage sludge, Fe- and Mn-humate decreasedplant Mo contents by 60, 40 and 75% in thegreenhouse. In the field study, the effects weresmaller, but Mo export through plant harvest increasedby 88% after P-fertilization and 84% aftervermiculite amendment. Mn-humate and sewage sludgedecreased plant Mo content by 25 and 40%,respectively, rendering the forage suitable forfeeding of ruminants. P-fertilization and vermiculitemay thus be recommended for the severely contaminatedsites to enhance phytoremediation through Mo export,and Mn-humate and sewage sludge application appearsuited to remediate the less severely contaminated sites.     

Bacterial culture of septic synovial structures of horses: Does a positive bacterial culture influence prognosis?

Reasons for performing study: The influence of synovial fluid culture on short‐ and long‐term prognosis of cases with septic synovitis requires study. Hypotheses: Horses with a positive bacterial culture from septic synovial fluid are less likely to survive or return to successful athletic function than those with a negative bacterial culture from septic synovial fluid. Methods: Records of mature horses presented to 2 equine referral hospitals for investigation of suspected septic synovitis were examined. Horses (n = 206) were included in the study if synovial fluid was submitted for full laboratory examination, including bacterial culture. A diagnosis of septic synovitis was based on a nucleated cell count >30 × 10⁹ cells/l or >90% neutrophils and other clinical, cytological and bacteriological parameters. Long‐term follow‐up was obtained by telephone questionnaire. Univariate analysis, using the Fisher's exact test, was used for all outcomes. Results: Fourteen (20.9%) of 67 horses with a positive bacterial culture from synovial fluid were subjected to euthanasia because of persistent synovial sepsis compared to 2 (1.44%) of 139 with negative bacterial cultures (P<0.001). Overall survival and successful long‐term return to function in horses with a positive bacterial culture was 50% (24/48 horses) compared to 70.5% (74/105) in culture negative horses (P = 0.01). In horses that survived to be discharged, successful long‐term return to function was not significantly different between culture positive and culture negative groups. Growth of Staphylococcus aureus from synovial fluid did not affect short‐term survival to discharge from the hospital compared to other positive bacterial culture; however, successful long‐term return to function was only 30.4% (4/13) in horses from which S. aureus was cultured compared to 73.9% (17/23) of horses in which other bacteria were cultured (P = 0.015). Conclusions and potential clinical relevance: Horses with a positive bacterial culture from a septic synovitis have a poorer prognosis for survival to discharge from hospital and overall long‐term return to function than horses that yielded no bacterial growth. When S. aureus was cultured, the long‐term prognosis was poorer.     

Effects of intravesical hydrostatic pressure and volume on the distensibility of the canine prostatic portion of the urethra

Positive-contrast retrograde urethrocystograms were obtained serially on 12 male dogs weighing 11.4 to 23.2 kg before, during, and after the injection of contrast medium until the urinary bladder neck and prostatic and membranous portions of the urethra remained open and distended as viewed by fluoroscopy. Correlations of intravesical volumes and pressures required to achieve maximum distension of the midprostatic portion of the urethra with body weight and surface area were not significant. Because of the variability in intravesical volumes and pressures encountered at maximum distension of the prostatic portion of the urethra, a dose of contrast material expressed relative to body weight or surface area could not be determined for consistently providing maximum distension of the prostatic portion of the urethra.     

Effects of transportation on early embryonic death in mares

Incidence of early embryonic death (EED) and associated changes in serum cortisol, progesterone and plasma ascorbic acid (AA) in transported mares were investigated. Mares were transported for 472 km (9 h) during either d 16 to 22 (T-3 wk, n = 15) or d 32 to 38 (T-5 wk, n = 15) of gestation. Blood samples were drawn from control, nontransported mares (NT-3 wk, NT-5 wk, n = 24) and transported mares pre-trip, midtrip, and at 0, 12, 24, 48 and 72 h post-transport and daily for the next 2 wk. Incidence of EED between transported and nontransported mares was not different (P greater than .05). Serum cortisol in all transported mares increased (P less than .05) relative to pre-trip values at midtrip and 0 h post-transport. Relative to NT mares, serum cortisol was higher (P less than .05) at midtrip in T-3 wk mares and 0 h post-transport in T-5 wk mares. Serum progesterone in all T mares increased (P less than .05) at midtrip relative to pre-trip values and was higher (P less than .05) in T-3 wk mares than in NT-3 wk mares at midtrip and 0 h post-transport. Post-transport decreases (P less than .05) in concentrations of progesterone were observed in mares that aborted. Plasma AA in transported mares increased (P less than .05) at midtrip in T-5 wk mares and decreased (P less than .05) relative to pre-trip values at 24 and 48 h post-transport (T-3 wk and T-5 wk mares, respectively).(ABSTRACT TRUNCATED AT 250 WORDS)     

The purification and characterisation of cervine IgM and IgG

A procedure is described for the isolation of immunoglobulin G (IgG) and immunoglobulin M (IgM) from hyperimmune cervine serum. Hybrids of red deer (Cervus elaphus) and wapiti (Cervus canadensis) were immunised with keyhole limpet hemocyanin (KLH). An immunoglobulin-containing fraction was precipitated from the hyperimmune serum using ammonium sulphate. The antigen-specific immunoglobulins were purified by KLH-conjugated sepharose affinity chromatography and further separated into IgM and IgG by gel-filtration chromatography. Purified immunoglobulin was analysed by polyacrylamide gel electrophoresis and isoelectric focusing. The molecular weights and isoelectric points of the composite chains of cervine IgG and IgM are presented.