Antimicrobial Activity of Culture Filtrate of Bacillus amyloliquefaciens RC-2 Isolated from Mulberry Leaves

ABSTRACT A potential antagonist, Bacillus amyloliquefaciens strain RC-2, against Colletotrichum dematium, mulberry anthracnose fungus, was obtained from healthy mulberry leaves by in vitro and in vivo screening techniques. Application of culture filtrate of RC-2 inhibited disease on mulberry leaves, indicating that suppression was due to antifungal compounds in the filtrate. Development of mulberry anthracnose on mulberry leaves was inhibited only when the culture filtrate was applied before fungal inoculation, and it was not inhibited by application after inoculation. These results suggest that the antifungal compounds in the filtrate exhibit a preventive effect on the disease. Peptone significantly increased production of the antifungal compounds. The culture filtrate of RC-2 also inhibited the growth of several other phytopathogenic fungi and bacteria, such as Rosellinia necatrix, Pyricularia oryzae, Agrobacterium tumefaciens, and Xanthomonas campestris pv. campestris, in vitro. From the culture filtrate of RC-2, seven kinds of antifungal compounds were isolated by high performance liquid chromatography analysis, and one of the compounds was determined as iturin A2, a cyclic peptide, by nuclear magnetic resonance and fast atom bombardment mass analysis.     

Investigating the selectivity of weed harrowing with new methods

In six field experiments on post‐emergence weed harrowing in spring barley, the effects of row spacing, timing, direction and orientation on crop/weed selectivity were investigated. The efficacies of increasing intensities of harrowing generated either by increasing number of passes or increasing driving speed were also tested. Selectivity was defined as the relationship between crop burial in soil immediately after treatment and weed control. To estimate crop burial, digital image analysis was used in order to make objective estimations. The study showed that narrow row spacing decreased selectivity in a late crop growth stage, whereas row spacing in the range 5.3–24 cm had no effects at an early growth stage. Harrowing across rows decreased selectivity in one out of two experiments. Whether repeated passes with the harrow were carried out in the same orientation along the rows or in alternative orientations forth and back was unimportant. There were indications that a high harrowing intensity produced by a single pass at high speed gave a lower selectivity than a similar intensity produced by several passes at a low speed. Impacts on selectivity, however, were small and only significant at high degrees of weed control. Timing had no significant impact on selectivity.     

Selenium, vitamin E and polyunsaturated fatty acid concentrations in goat kids with and without nutritional myodegeneration

Mean selenium and vitamin E (alpha-tocopherol) concentrations in the liver, myocardium and skeletal muscle of nine goat kids with nutritional myodegeneration (NMD) were significantly lower than those in a group of nine kids dying from other causes. Each of the 18 kids was from a different property in the southern half of the North Island. The polyunsaturated fatty acid composition of the liver, myocardium and skeletal muscle was not significantly different between the two groups, but the overall level of peroxidisable polyunsaturated fatty acids appeared high in both groups. A further 21 kids with confirmed NMD and live 'controls', submitted as routine cases to Palmerston North Animal Health Laboratory, were included with the above 18 kids in a comparison of liver selenium and alpha-tocopherol concentrations. Kids with NMD had liver selenium concentrations ranging from 170-1100 (mean = 380) nmol/kg and alpha-tocopherol from 0.7-11.0 (mean =2.2) micromol/kg. In control kids, the liver selenium concentration ranged from 530-4300 (mean = 1220) nmol/kg and a-tocopherol from 1.7-14.0 (mean = 5.6) pmollkg. Although these ranges overlapped, the results suggest that liver selenium concentrations <500 nmol/kg and alpha-tocopherol concentrations <2.5 micromol/kg should be regarded as deficient. Liver selenium concentrations from 500-1100 nmol/kg and alpha-tocopherol concentrations from 2.5-10 micromol/kg should be considered marginal. Goat kids appear to require more selenium than lambs or calves which may explain the higher prevalence of NMD in kids. In some cases, however, the disease is associated with low alpha-tocopherol suggesting that supplementation with selenium alone may not always be sufficient.     

Globalization, international trade and animal health: the new roles of OIE

In order for countries and their stakeholders to maximize the benefits of globalization they must become familiar with, and must adhere to, the rights and obligations set out by the World Trade Organization (WTO) under the Agreement on Sanitary and Phytosanitary Measures (SPS). For the purpose of trade in animals and animal products, they must also adhere to the standards, guidelines and recommendations established by the World Organisation for Animal Health (OIE). Countries are also encouraged to participate in this standard setting process of the OIE. Only after implementing these requirements and after strengthening the veterinary infrastructures and their surveillance and monitoring systems, will countries be able to fully benefit from these new international trade rules.     

An outbreak of Newcastle disease in free-living pheasants (Phasianus colchicus).

The epidemiology of an outbreak of Newcastle disease in a population of approximately 12,000 free-living pheasants (Phasianus colchicus) on the island of Faen? in Denmark in 1996 is described. The mortality during the epizootic was 56%. The spread of the disease between 7 groups of pheasants could be demonstrated over an observation period of 3 weeks. A total of 70 avian paramyxovirus serotype 1 (APMV-1) isolates was made from the flock. The intra cerebral pathogenicity indices of the 4 isolates tested were in the range 1.78-1.88. By means of immunoperoxidase monolayer assay with murine monoclonal antibodies and sequence analysis of an RT-PCR amplified segment of the F0 viral protein it was found, that the virus belonged to the highly virulent C1 antigenic group and that the amino acid sequence at the F0 cleavage site corresponded with the sequences of virulent APMV-1 strains. Based on the epidemiological circumstances it is believed that the virus was transmitted to the pheasants by feral birds.     

In vitro expression and inhibition of procoagulant activity produced by bovine alveolar macrophages and peripheral blood cells

Local and systemic activation of coagulation is frequently associated with bacterial sepsis. The coagulopathy is due, at least in part, to expression of tissue factor (TF) by monocytes and macrophages. The purpose of this study was to evaluate the expression of procoagulant activity by bovine alveolar macrophages, leukocytes and platelets, and to determine the relative potency of three chemical inhibitors of TF expression (pentoxifylline, retinoic acid, and cyclosporin A). Bovine alveolar macrophages were stimulated with lipopolysaccharide (LPS) derived from Pasteurella haemolytica or recombinant bovine tumour nervous factor (TNF) and dose- and time-dependent effects on TF expression were studied. LPS and TNF induced TF expression in alveolar macrophages and LPS treatment of whole blood induced TF expression in mononuclear cells. Neutrophils and platelets also expressed procoagulant activity, but this activity was not inhibited by anti-bovine TF monoclonal antibody. Pentoxifylline (40 mol/L), retinoic acid (0.01 mmol/L) and cyclosporin A (0.08 mol/L) inhibited TF expression when added concurrently with LPS or TNF, but not when added 4 h after stimulation. TF mRNA was not detected in unstimulated alveolar macrophages by Northern blot analysis. In contrast, exposure to LPS or TNF for 6 h induced marked expression of TF mRNA, which was inhibited by treatment with pentoxifylline, retinoic acid and cyclosporin A. Expression of TNF by alveolar macrophages stimulated with LPS was also inhibited by these compounds. Our results indicate that procoagulant activity expressed by alveolar macrophages and monocytes is associated with expression of TF, whereas procoagulant activity expressed by neutrophils and platelets is not. The concentrations of pentoxifylline and retinoic acid necessary for inhibition of TF expression in vitro may not be achievable in vivo owing to their toxic effects. However, the in vitro concentration of cyclosporin A that inhibited TF expression did not exceed the plasma concentration observed in humans, and therefore may be useful for inhibition of TF expression in vivo.Abbreviations BAL bronchoalveolar lavage - LPS lipopolysaccharide - cDNA cloned deoxyribonucleic acid - cAMP cyclic adenosine monophosphate - GAPDH glyceraldehyde phosphate dehydrogenase - mRNA messenger ribonucleic acid - TF tissue factor - TNF tumour necrosis factor - DPBS Dulbecco's phosphate-buffered saline     

Effect of endocytic and metabolic inhibitors on the internalization and intracellular growth of Brucella abortus in Vero cells.

Uptake, transfer to rough endoplasmic reticulum, and intracellular growth of Brucella abortus were studied in Vero cells treated with endocytic and metabolic inhibitors. Infection of Vero cells was suppressed when inhibitors of energy metabolism (iodoacetate, dinitrophenol), receptor-mediated endocytosis (monodansylcadaverine, amantadine, methylamine), or endosomal acidification (chloroquine, ammonium chloride, monensin) were added to the inoculum. Inhibition was not observed when these drugs were added after the inoculation period. Infection of Vero cells by B abortus was inhibited by dibutyryl-cyclic adenosine monophosphate and Vibrio cholerae enterotoxin, but was stimulated by dibutyryl-cyclic guanosine monophosphate and escherichia coli heat-stable enterotoxin a. Uptake of B abortus by Vero cells was not prevented by colchicine, but was abolished by cytochalasin B. Uptake of heat-killed B abortus and noninvasive E coli was similar to that of viable brucellae. Intracellular growth of B abortus was not affected by cycloheximide. Results indicate that: B abortus may be internalized by a receptor-mediated phagocytic process; transfer of B abortus from phagosomes to rough endoplasmic reticulum may require endosomal acidification; and replication of B abortus within the rough endoplasmic reticulum may not depend on protein synthesis by the host cell.     

Fibroblast growth factor-10 maintains the survival and promotes the growth of cultured goat preantral follicles

The aim of the present study was to investigate the effects of fibroblast growth factor-10 (FGF-10) on the survival, activation (transition from primordial to primary follicles), and growth of goat preantral follicles cultured in vitro. Pieces of ovarian cortex were cultured for 1 and 7 d in the absence or presence of FGF-10 (0, 1, 10, 50, 100, and 200 ng/mL). Noncultured and cultured tissues were processed and analyzed by histology, transmission electron microscopy, and viability testing. Results showed that after 7 d, a greater percentage (79.9%) of morphologically normal follicles (containing an oocyte with regular shape and uniform cytoplasm, and organized layers of granulosa cells without a pyknotic nucleus) was observed when cultured with 50 ng/mL of FGF-10 when compared with other concentrations of FGF-10 (0 ng/mL, 67.3%; 1 ng/mL, 68.2%; 10 ng/mL, 63.3%; 100 ng/mL, 64.4%; 200 ng/mL, 52.7%). Ultrastructural analyses and viability testing using fluorescent markers confirmed the follicular integrity of FGF-10 (50 ng/mL)-treated fragments after 7 d of culture. After 7 d, all FGF-10 concentrations reduced the percentage of primordial follicles and increased the percentage of developing follicles. In the presence of 50 ng/mL of FGF-10, follicles increased in diameter after 7 d of culture when compared with other concentrations tested. In conclusion, this study demonstrates that FGF-10 maintains the morphological integrity of goat preantral follicles and stimulates the growth of activated follicles in culture. The culture conditions identified here contribute to the understanding of the factors involved in goat early follicular development.