Fusarium moniliforme secretes four endopolygalacturonases derived from a single gene product

Extracellular endopolygalacturonase, purified from the pathogenic fungus Fusarium moniliforme, consists of four molecular forms (38, 41·5, 45, and 48·5 kDa, respectively. Three forms (38, 41·5, and 45 kDa) were purified to homogencity by FPLC on a Mono S column followed by electroelution after SDS-PAGE. The N-terminal amino acid sequences of each of the three forms, and of a mixture containing all four forms were shown to be identical to that predicted from the nucleotide sequence of the endopolygalacturonase gene previously cloned from F. moniliforme. Enzymatic deglycosylation experiments revealed the presence of N-linked, high mannose oligosaccharide side-chains on all four forms of endo polygalacturonase. Hydrogen fluoride catalysed chemical deglycosylation of the polygalacturonase mixture yielded a single polypeptide with an apparent molecular mass of 36·2 kDa. Southern blot analysis, carried out at high stringency with an endopolygalacturonase-specific probe on genomic DNA digested with three different restriction enzymes, showed a single hybridizing restriction fragment in all three digests. A single 2·0 Mb chromosome hybridized with the endo polygalacturonase-specific probe, as shown by Southern blot analysis of F. moniliforme chromosomes separated by CHEF electrophoresis. Northern blot analysis revealed only one mRNA species 1350 nt encoding endo polygalacturonase. These data indicate that a single gene encodes the endopolygalacturonases of F. moniliforme.     

Bacterial biodegradation of ethoxylated surfactants

Agricultural, industrial and domestic use of surfactants leads to the entry of these compounds into terrestrial and aquatic ecosystems. Synthetic surfactants vary significantly in structure, but most consist of alkyl or alkylphenol groups attached to nonionic or anionic hydrophilic moieties. Continued use of these compounds is usually justified on the basis that they do not cause pollution problems because they undergo biodegradation by micro-organisms present in soils and surface waters. In accomplishing biodegradation, micro-organisms, predominantly bacteria, are exploiting these potentially useful resources of reduced carbon to derive energy and support growth in situations which are otherwise frequently oligotrophic. This paper reviews aspects of surfactant biodegradation, especially in relation to alcohol and alkylphenol ethoxylates used extensively as adjuvants for agrochemicals. In principle, bacteria can employ two strategies to gain access to the aliphatic chains in alcohol ethoxylate surfactants: separation of the hydrophobic chain from the hydrophile (central fission), or direct attack on the -terminal of the alkyl chain of the intact surfactant. Direct exo-cleavage of ethylene glycol units from the polyethylene glycol (PEG) chain also provides a third route to assimilable carbon. In pure cultures of known degraders or in mined environmental samples, all three strategies are exploited, some even within the same organism. Central fission occurs predominantly at the alkyl-ether bond, but may also occur within the PEG chain itself, thus producing various glycol intermediates which accumulate in pure cultures but appear only transiently in mixed environmental samples. Against this background, the relative resistance of some alkylphenol ethoxylates to biodegradation can be assessed in mechanistic terms. The steric bulk of the aryl nucleus effectively eliminates the central fission pathway. Moreover, some alkyl phenol ethoxylates contain branched alkyl chains which restrict ω-β-oxidation. As a result, ethoxylate shortening appears to be the major course of biodegradation observed so far. Not surprisingly, these surfactants are observed to undergo extensive primary biodegradation (removal of surfactant properties) but relatively restricted ultimate degradation to carbon dioxide and normal cell components.     

The Danish Integrated Farm Management System—sector module for plant production1

The Danish Integrated Farm Management System is a PC-based system developed by the Danish agricultural advisory service covering all management and control aspects of a farm. It consists of a joint module linking a number of sector modules (cattle production, plant production, economy, pig production and buildings/machinery). The sector module for plant production provides facilities for detailed planning, decision support and control, including all subjects of a complete cultivation plan for each individual field. The fertilizer plan and the crop protection plan provide detailed specifications within the overall cultivation plan. For decision support there are advanced sub-modules for fertilization, crop protection and grassland management. PC-Plant Protection is an optional and fully integrated sub-module of The Integrated Farm Management System, or a self-contained program product.     

Field evaluation of the single intradermal cervical tuberculin test and the interferon-gamma assay for detection and eradication of bovine tuberculosis in Spain

A field comparison of the interferon-gamma (IFN-gamma) assay and the single intradermal cervical tuberculin (SICT) test for the diagnosis of bovine tuberculosis was conducted. A total of 1136 cattle belonging to 85 herds placed in 'Castilla y León' (northwestern Spain) were chosen, and 21 of these herds were subjected to the diagnostic assays two or three times at intervals of at least 4 months. All the animals positive to any of the tests were slaughtered and tuberculosis was confirmed by culture isolation method (CIM) and further identification by means of PCR. Only 10.6% of cattle reacted with the bovine PPD in the SICT test, a percentage that increased to 12.8% in the IFN-gamma assay. The sensitivity of the IFN-gamma assay compared to CIM was shown to be higher (84.9%) than that of the SICT test (80.2%), but the combination of both tests offered the highest sensitivity (92.9%). The number of false positive reactors (those animals in which CIM was negative) was considerably higher for the IFN-gamma assay than for the SICT test and, conversely, the number of false negative animals (M. bovis isolation but negative immunological result) was higher for the skin test than for the interferon assay. In the herds tested twice, tuberculosis was eradicated after the second cycle of testing in 50%, and in 75% after the third cycle in herds tested three times. The combination of these two techniques instead of separately seems, therefore, to be useful in eradication programmes against bovine tuberculosis.     

Identification of aesculin-hydrolyzing streptococci, lactococci, aerococci and enterococci from subclinical intramammary infections in dairy cows

Aesculin-hydrolyzing, catalase-negative, gram-positive cocci isolated from subclinical intramammary infections in dairy cows were identified to species level using growth characteristics and biochemical activity. The results indicated that the aesculin-hydrolyzing cocci associated with this type of infection are a very heterogenic group. S. uberis strains, including inulin- or beta-glucuronidase-negative isolates, accounted for only about one-third of the collection, and Enterococcus faecalis strains for one-fifth. Other species of some importance included (in descending order of isolation frequency) Aerococcus viridans, Streptococcus pluranimalium, Lactococcus garvieae, Streptococcus bovis and Streptococcus gallolyticus.     

Paraneoplastic thrombocytosis-induced systemic thromboembolism in a cat

A six-year-old cat presented with clinical signs consistent with distal aortic thromboembolism while clinical signs of cardiovascular disease were absent. Diagnostics, including thoracic radiographs, electrocardiography, and echocardiography revealed no cardiovascular anomalies. Thoracic radiographs revealed multifocal pulmonary lesions consistent with neoplasia. Complete blood cell count demonstrated a marked thrombocytosis, leukopenia, and neutropenia. Histopathology of the pulmonary lesions confirmed multiple bronchoalveolar carcinomas. Myelodysplasia with megakaryocytic hyperplasia and ineffective myelopoiesis was noted on bone-marrow histopathology from multiple sites. The absence of other causes suggested a paraneoplastic thrombocytosis. The diagnosis of paraneoplastic thrombocytosis-induced thromboembolism was made due to the lack of underlying cardiac disease and the presence of a marked thrombocytosis. The presence of thrombocytosis and thromboembolism associated with neoplasia is discussed.     

Studies on the transmission of Theileria annulata to cattle by the tick Hyalomma lusitanicum

The role of the ixodid tick Hyalomma lusitanicum Koch 1844 as a vector of Mediterranean or tropical theileriosis (caused by the protozoan parasite Theileria annulata Dschunkowsky et Luhs 1904) in southern Spain was studied. Hyalomma lusitanicum was the most common tick, and the only species of the genus Hyalomma L., found on T. annulata-infected cattle from the theileriosis enzootic area studied (province of Cádiz, southern Spain). Likewise, we found that all sera of the cattle previously considered as suspected of theileriosis by clinical signs, tested for T. annulata antibodies, were positive and all blood samples of these suspected cattle examined had infected erythrocytes. Partially fed H. lusitanicum adults were collected in the field on T. annulata-infected cattle in this enzootic area and fed on an uninfected calf in an experimental farm free of theileriosis and ticks. At approximately 3 weeks post-tick feeding on the calf, this became positive for T. annulata antibodies and T. annulata merozoites were found in erythrocytes from blood smears. These results show the ability of H. lusitanicum to transmit the protozoan parasite T. annulata to susceptible cattle and indicate that H. lusitanicum is probably an important vector of T. annulata in the enzootic area surveyed.     

Aggregation of sow lactobacilli with diarrhoeagenic Escherichia coli

A total of 20 strains of lactobacilli were isolated from the oesophagus and vagina of 20 sows at the time of partus. Aggregation activity was seen between six homofermentative autoaggregative lactobacilli and three strains of pathogenic Escherichia coli with F4, F5 and F6 fimbriae. The highest aggregation activity was observed between vaginal Lactobacillus acidophilus PV 32 or oesophageal OE 2/1 and E. coli with F4 (K88). The presence of aggregation-promoting factor (APF) was confirmed by polymerase chain reaction (PCR) with primers of a specific fragment the apf gene derived from human L. gasseri 4B2 in one oesophageal L. acidophilus strain OE 2/1. We propose that autoaggregative lactobacilli that aggregate with diarrhoeagenic E. coli can express a class of APF proteins that exhibit the function of an aggregation mediator.     

Histological Aspects of a Hypersensitive Response in Poplar to Melampsora larici-populina

ABSTRACT The course of the infection and development of the biotrophic fungus Melampsora larici-populina on leaf tissue from the hybrid poplar Populus deltoides x P. nigra 'Ogy' was monitored at the histological level. Leaf disks were inoculated with one of two rust physiological races (E1 and E2), resulting in interactions that were either incompatible (race E1) or compatible (race E2). In the compatible interaction, the fungus rapidly colonized the leaf without inducing any apparent host response. Symptoms appeared on the leaf several days after inoculation just prior to spore dissemination. The incompatible interaction was characterized by the early collapse and disorganization of cytoplasm of infected cells 17 h after inoculation and within 2 h after the appearance of the first haustoria. Resistance to M. larici-populina was mediated through a hypersensitive response, since it was extremely localized and involved only the few cells that were in the immediate vicinity of each infected cell.