Maceration of plant tissue by fungi is inhibited by recombinant antipectinase antibodies

Polyclonal antiserum from mice immunized with extracellular proteins from Rhizoctonia solani inhibited pectinase and cellulase activities in cell free culture supernatants of Rhizoctonia solani. Spleen mRNA from these mice was used to construct a cDNA library from which antipectinase ScFv antibodies were isolated using phage display techniques. Soluble ScFv antibodies produced by individual clones in Escherichia coli inhibited polygalacturonase in the culture supernatants of a range of fungal pathogens, including ascomycetes, basidiomycetes, and oomycetes. The soluble antibodies also inhibited maceration of potato tissue by these pathogens.     

Antagonistic rhizoplane bacteria induce diverse morphological alterations in Peronosporomycete hyphae during <Emphasis Type="Italic">in vitro</Emphasis> interaction

A total of 150 bacteria were isolated from rhizoplanes of the host and non-host plants of a phytopathogenic Peronosporomycete Aphanomyces cochlioides. Upon screening, 5% of the isolates were evaluated as antagonists as they inhibited radial growth of A. cochlioides AC-5 hyphae in a dual culture assay. In addition, those antagonistic bacteria also induced characteristic morphological alterations in the A. cochlioides AC-5 hyphae that grew towards bacterial colonies. Hyphal morphological alterations observed in AC-5 and other tested strains of Peronosporomycetes included excessive branching, curly growth, unusually longer and pointed tip formation and swelling; all of these were comparable to the alterations induced by known antimicrobial compounds. Among the antagonistic bacteria, Pseudomonas sp. strain EC-S101 induced a unique branching pattern (tree-like) in AC-5 hyphae by continuous apical bifurcation of successive hyphae, where increases in number of branches and hyphal area were linearly correlated with time up to 10 h. Our observations suggested that the pathogen might have lost its ability of normal branch production; however maintained the capability of self-branching. Soluble extracts from the culture fluids of Pseudomonas sp. strain EC-S101 and Stenotrophomonas maltophilia EC-S105 induced similar excessive branching and curly growth in A. cochlioides hyphae as the respective bacterium. These results revealed that bacterial metabolites appeared to be responsible for induction of morphological alterations. Interestingly, the antagonistic bacteria that induced hyphal morphological alterations, also efficiently suppressed in vivo damping-off disease caused by AC-5. We suggest that antagonistic rhizoplane bacteria have the capability to induce diverse morphological alterations in Peronosporomycetes hyphae during in vitro interactions. Hyphal morphological alterations associated with growth inhibition and the induction of characteristic morphological changes indicate antagonistic activity against the Peronosporomycete.     

Molecular characterization of the major outer membrane protein of Haemophilus somnus

The major outer membrane protein (MOMP) of Haemophilus somnus shows antigenic and molecular mass diversity that forms the basis of a preliminary grouping system for H. somnus strains. In this study, the gene encoding MOMP of H. somnus strain 8025 was cloned in three overlapping fragments by PCR techniques, and then sequenced. The gene consists of a 1164-bp open reading frame encoding a deduced 380-amino acid protein with a 19-amino acid signal sequence, giving a mature protein with a calculated molecular mass of 39,913 Da. Significant homology was found between MOMP and porin protein sequences of bacteria in Pasteurellaceae species. When expressed in Escherichia coli, the protein from the MOMP gene directed by the T7 promoter was identical in size (approximately 40 kDa) to native MOMP and reacted with MOMP-specific antibodies. Comparisons of the MOMP gene sequences from six unrelated strains of H. somnus to that of strain 8025 revealed that the genes of three MOMP type 1 strains were highly conserved with that of strain 8025 in length and sequence. However, two MOMP type 3c strains and one MOMP type 3a strain differed markedly from the MOMP of strain 8025 in their 3'-terminal halves. Their deduced MOMP amino acid sequences differed in sequence (3c, 80.5 and 82.7% identity; 3a, 62.4% identity) and in length (3c, 384 and 376; 3a, 316), indicating that the molecular differences are the basis of antigenicity and molecular mass differences of H. somnus MOMP. In the predicted MOMP secondary structure, the variable sequences primarily mapped to putative surface-exposed loops, and a variable and surface-exposed epitope of MOMP-specific antibody was identified in the seventh-largest loop. These findings are useful for understanding the structural and immunological characteristics of H. somnus.     

Rice Pi-ta gene Confers Resistance to the Major Pathotypes of the Rice Blast Fungus in the United States

ABSTRACT The Pi-ta gene in rice prevents the infection by Magnaporthe grisea strains containing the AVR-Pita avirulence gene. The presence of Pi-ta in rice cultivars was correlated completely with resistance to two major pathotypes, IB-49 and IC-17, common in the U.S. blast pathogen population. The inheritance of resistance to IC-17 was investigated further using a marker for the resistant Pi-ta allele in an F(2) population of 1,345 progeny from a cross of cv. Katy with experimental line RU9101001 possessing and lacking, respectively, the Pi-ta resistance gene. Resistance to IC-17 was conferred by a single dominant gene and Pi-ta was not detected in susceptible individuals. A second F(2) population of 377 individuals from a reciprocal cross between Katy and RU9101001 was used to verify the conclusion that resistance to IC-17 was conferred by a single dominant gene. In this cross, individuals resistant to IC-17 also were resistant to IB-49. The presence of Pi-ta and resistance to IB-49 also was correlated with additional crosses between 'Kaybonnet' and 'M-204', which also possess and lack Pi-ta, respectively. A pair of primers that specifically amplified a susceptible pi-ta allele was developed to verify the absence of Pi-ta. We suggest that Pi-ta is responsible for resistance to IB-49 and IC-17 and that both races contain AVR-Pita genes.     

Inhibitory effect of pyrimidine and purine nucleotides on the multiplication of Babesia gibsoni: possible cause of low parasitemia and simultaneous reticulocytosis in canine babesiosis

The present study was conducted to determine the cause of low parasitemia and simultaneous reticulocytosis in canine babesiosis. The parasitemia was significantly decreased in in vitro cultures of Babesia gibsoni by the pretreatment of host canine erythrocytes with lead acetate, which is a specific inhibitor of pyrimidine 5'-nucleotidase subclass I (P5N-I). The serum from dogs chronically infected with B. gibsoni did not decrease the activities of hexokinase, glucose-6-phosphate dehydrogenase or 6-phosphogluconate dehydrogenase in canine reticulocytes, although it was previously reported that this serum had inhibitory effects on both the maturation of reticulocytes and the canine P5N-I and purine-specific 5'-nucleotidase activities. Furthermore, the in vitro multiplication of B. gibsoni was significantly inhibited by pyrimidine nucleotides such as cytidine 5'-monophosphate (5'-CMP), which is preferentially catalyzed by P5N-I and also inhibits the morphological maturation of canine reticulocytes. Purine nucleotides such as inosine 5'-monophosphate (5'-IMP) also had an inhibitory effect on the multiplication of this parasite. These results suggest that nucleotides such as 5'-CMP and 5'-IMP might accumulate in young erythrocytes and/or serum in dogs infected with B. gibsoni as a result of the decreased activity of erythrocyte 5'-nucleotidase, and the accumulation of these nucleotides might inhibit the multiplication of this parasite and simultaneously retard the maturation of reticulocytes. The results obtained from the in vitro examinations in the present study may partially clarify the relationship between low parasitemia and simultaneous reticulocytosis in vivo in canine babesiosis.     

Anti-diabetic potential of ursolic acid stearoyl glucoside: a new triterpenic gycosidic ester from Lantana camara

A new stearoyl glucoside of ursolic acid, urs-12-en-3β-ol-28-oic acid 3β-d-glucopyranosyl-4′-octadecanoate and other compounds were isolated from the leaves of Lantana camara L. The structure of this new glycoside was elucidated and established by standard spectroscopic methods. In streptozotocin-induced diabetic rats it showed significant reduction in blood glucose level.     

Aleutian mink disease virus in furbearing mammals in Nova Scotia,Canada

Background

Aleutian mink disease virus (AMDV) is widespread among ranched and free-ranging American mink in Canada, but there is no information on its prevalence in other wild animal species. This paper describes the prevalence of AMDV of 12 furbearing species in Nova Scotia (NS), Canada.

Methods

Samples were collected from carcasses of 462 wild animals of 12 furbearing species, trapped in 10 NS counties between November 2009 and February 2011. Viral DNA was tested by PCR using two primer pairs, and anti-viral antibodies were tested by counterimmunoelectrophoresis (CIEP) on spleen homogenates.

Results

Positive PCR or CIEP samples were detected in 56 of 60 (93.3%) American mink, 43 of 61 (70.5%) short-tailed weasels, 2 of 8 (25.0%) striped skunks, 2 of 11 (18.2%) North American river otters, 9 of 85 (10.6%) raccoons, and 2 of 20 (10.0%) bobcats. Samples from six fishers, 24 coyotes, 25 red foxes, 58 beavers, 45 red-squirrels and 59 muskrats were negative. Antibodies to AMDV were detected by CIEP in 16 of 56 (28.6%) mink and one of the 8 skunks (12.5%). Thirteen of the mink were positive for PCR and CIEP, but three mink and one skunk were CIEP positive and PCR negative. Positive CIEP or PCR animals were present in all nine counties from which mink or weasel samples were collected.

Conclusions

The presence of AMDV in so many species across the province has important epidemiological ramifications and could pose a serious health problem for the captive mink, as well as for susceptible wildlife. The mechanism of virus transmission between wildlife and captive mink and the effects of AMDV exposure on the viability of the susceptible species deserve further investigation.     

Impacts of Municipal Wastewater on The Transport Characteristics of Reactive Solutes Through Agricultural Soils

Reliable transport parameters of agrochemicals and soluble pollutants are crucial for modeling and management of soil and groundwater quality. This study investigated impacts of municipal wastewater on the transport parameters of five heavy metal/metalloid compounds (NaAsO₂, Cd(NO₃)₂, Pb(NO₃)₂, Ni(NO₃)₂ & ZnCl₂), two pesticides (cartap & carbendazim) and an inert salt (CaCl₂) in four agricultural soils of Bangladesh. Solute-breakthrough concentrations were measured in repacked soil columns with time-domain reflectometry (TDR) both before and after wastewater treatment. Transport velocity (V), dispersion coefficient (D), dispersivity (λ) and retardation factor (R) of the solutes, and pertinent soil properties were determined. Wastewater reduced bulk density (γ) of the soils (from 1.32–1.37 g/cm³ to 1.26–1.35 g/cm³) by increasing organic carbon (OC) (from 0.37%–0.84% to 0.40–0.93%), increased pore-size distribution index (n) (by 0.02 unit) and reduced soil pH (from 6.32–7.45 to 5.92–6.46). D and λ decreased while V and R increased after wastewater treatment; D decreased and R increased linearly with decreasing bulk density. The correlations of V, D, and R with n improved significantly (p < 0.05) after wastewater treatment. The correlation between λ and OC improved markedly for Ca, Pb, Ni, and cartap. The observed indicative results have practical implications in developing pedo-transfer functions for solute-transport parameters using basic soil properties, which are subject to progressive modification due to agrochemicals application and wastewater irrigation.