Soil organic matter fractionation as a tool for predicting nitrogen mineralization in silty arable soils

After decades of searching for a practical method to estimate the N mineralization capacity of soil, there is still no consistent methodology. Indeed it is important to have practical methods to estimate soil nitrogen release for plant uptake and that should be appropriate, less time consuming, and cost effective for farmers. We fractionated soil organic matter (SOM) to assess different fractions of SOM as predictors for net N mineralization measured from repacked (disturbed) and intact (undisturbed) soil cores in 14 weeks of laboratory incubations. A soil set consisting of surface soil from 18 cereal and root‐cropped arable fields was physically fractionated into coarse and fine free particulate OM (coarse fPOM and fine fPOM), intra‐microaggregate particulate OM (iPOM) and silt and clay sized OM. The silt and clay sized OM was further chemically fractionated by oxidation with 6% NaOCl to isolate an oxidation‐resistant OM fraction, followed by extraction of mineral bound OM with 10% HF (HF‐res OM). Stepwise multiple linear regression yielded a significant relationship between the annual N mineralization (kg N/ha) from undisturbed soil and coarse fPOM N (kg N/ha), silt and clay N (kg N/ha) and its C:N ratio (R² = 0.80; P < 0.01). The relative annual N mineralization (% of soil N) from disturbed soils was related to coarse fPOM N, HF‐res OC (% of soil organic carbon) and its C:N ratio (R² = 0.83; P < 0.01). Physical fractions of SOM were thus found to be the most useful predictors for estimating the annual N mineralization rate of undisturbed soils. However, the bioavailability of physical fractions was changed due to the disturbance of soil. For disturbed soils, a presumed stable chemical SOM fraction was found to be a relevant predictor indicating that this fraction still contains bio‐available N. The latter prompted a revision in our reasoning behind selective oxidation and extraction as tools for characterizing soil organic N quality with respect to N availability. Nonetheless, the present study also underscores the potential of a combined physical and chemical fractionation procedure for isolating and quantifying N fractions which preferentially contribute to bulk soil N mineralization. The N content or C:N ratio of such fractions may be used to predict N mineralization in arable soils.     

Nutritional Enhancement of Biocontrol Activity of Candida sake (CPA-1) against Penicillium expansum on Apples and Pears

Pome fruits are poor in nitrogenous compounds and the addition of nitrogen can improve colonisation of the fruits by antagonists. Twenty-two nitrogenous compounds were evaluated for their effect on Candida sake (CPA-1) growth in vitro. Ten compounds that induced greater growth were applied with the antagonist to wounded fruits to evaluate their effect on enhancing control of Penicillium expansum. Calcium chloride and 2-deoxy-D-glucose were also tested. L-serine and L-aspartic acid enhanced biocontrol by C. sake against P. expansum on apples. On apples and pears, ammonium molybdate, calcium chloride and 2-deoxy-D-glucose improved the capacity of the antagonist to control P. expansum. The addition of ammonium molybdate at 1 mM allowed C. sake to be used on apples and pears at a lower concentration without diminishing control. Similar results were observed with the addition of calcium chloride to the antagonist. 2-deoxy-D-glucose at 6 and 18 mM enhanced biocontrol on pears by over 81%, but on apples the improvement of biocontrol was observed only at 6 mM. In cold storage, the combination of ammonium molybdate and C. sake completely eliminated the incidence of blue mould on pears, and reduced its severity and incidence by more than 80% on apples.     

Effects of Azoxystrobin on Mycotoxin Production in a Carbendazim-Resistant Strain ofFusarium sporotrichioides

Carbendazim-resistant (RS) and control (CS) strains ofFusarium sporotrichioides Sherb., previously developed in our laboratory, were exposed to graded concentrations of azoxystrob in in broth media under shake-culture conditions for 2, 3, 4 and 8 days. Azoxystrobin concentrations were 0, 1, 10 and 100 mg 1^-1 broth and cultures were incubated at a constant 25°C. Mycelial growth was significantly affected by strain (P<0.01), azoxystrobin concentration (P<0.001) and incubation time (P<0.001). Combined results for the four incubation times showed that CS yielded higher mycelial mass than RS (P<0.01) only in the absence of azoxystrobin. At fungicide additions of 1, 10 and 100 mg P^-1 mycelial growth was reduced (P<0.001) with minimal strain differences (P>0.05) at all three doses of azoxystrobin. Significant (P<0.05 or better) strain-fungicide interactions were recorded in trichothecene production following exposure to azoxystrobin. At 4 and 8 days of incubation, the 10 mg 1^-1 addition of azoxystrobin stimulated T-2 toxin synthesis (P<0.05) only in RS cultures. In contrast, T-2 toxin enhancement in CS cultures occurred only on day 8 but at a lower level of azoxystrobin (1 mg1^-1). Thus, the stimulation of T-2 toxin synthesis depended upon strain and azoxystrobin level. Production of diocetoxyscirpenol (DAS) was affected by a more complex set of interactions. Overall means showed that, in comparison with initial values (on day 2 or 3), DAS output maximized significantly(P<0.05) on day 4 in RS cultures and on day 8 in CS. Marked strain effects were observed on exposure to the 10 mg 1^-1 level of azoxystrobin. At this level, DAS production was enhanced in RS only after 4 (P<0.01 ) and 8 (P<0.05) days of incubation, while in contrast, CS reduced DAS production. As with T-2 toxin, DAS production in CS was stimulated (P<0.05 or better) only at low exposure levels of azoxystrobin. In the case of neosolaniol (NEO), however, the main effect of strain was significant (P<0.05), with CS producing consistently more of the mycotoxin than RS on day 4 of the experiment. At this point, the NEO:T-2 toxin ratio was also higher in CS (0.63) than in RS (0.12), a feature reported by us previously. In conclusion, the present investigation has shown for the first time that the development of resistance to one fungicide can affect trichothecene production inF. sporotrichioides on exposure to a second fungicide. These results have been incorporated into a new classification scheme for fungicide efficacy which is also presented in this paper. http://www.phytoparasitica.org posting Oct. 7,2001.     

Influence of age and purpose for testing on the cut-off selection of serological methods in bovine neosporosis

The aim of this study was to investigate the need for different cut-off points, according to animal age and the purpose of testing, for two of the most widely used serological techniques in bovine neosporosis, IFAT and a crude antigen ELISA (Civtest, HIPRA). Therefore, the population reference sera used were defined using a combination of multiple criteria such as epidemiological/clinical and histopathological parameters and an immunoblot test. Firstly, foetuses and breeding cattle (heifers and cows) were considered as separate subpopulations for serological evaluation. Secondly, cut-off points for each serological technique (IFAT and ELISA) according to age group (foetuses and breeding cattle) and the different practical applications (detection of infection and abortion) were calculated following the receiver operating characteristics (ROC) analysis. Cut-off points were defined, for IFAT and ELISA for aborted breeding cattle and for IFAT alone in the case of the foetuses, assuming an equivalent cost of false positive and negative results. In infected breeding cattle, for IFAT and ELISA and in foetuses for ELISA, two possible cut-off values were obtained, one for a maximum sensitivity and one for a maximum specificity and the intervals of unclear results were defined. In this case, a cut-off value for equal sensitivity and specificity was also estimated. When cut-off points for infected breeding cattle, 1:100-1:250 for IFAT and 0.306-0.451 for ELISA were applied to a target population, optimal and similar negative and positive predictive values together with similar apparent and true prevalence results were observed suggesting the possibility of using both tests interchangeably.     

Multispecies and multiple anthelmintic resistance on cattle nematodes in a farm in Argentina: the beginning of high resistance?

There are very few resistance records on cattle nematodes. South American successive records have been increasing rapidly since the year 2000. In Argentina, increasing dissatisfaction based on the exclusive use of macrocyclic lactones has prompted the use of benzimidazoles in the Pampean region. The studied farm is located in the Argentina humid Pampas and had apparently poor results after anthelmintic treatments. Evaluation of resistance was firstly based on faecal egg reduction after treatment in November 2001 and June 2002 and was complemented with worm counts obtained by necropsy in June 2002. The study reports that the reduction of faecal egg excretion after benzimidazole (reduction 31-79%) or ivermectin treatments (76-97%) in November 2001 was insufficient and these poor results were confirmed in June 2002 (benzimidazoles (65-89%) and ivermectin (47-77%)). Several methods for evaluating faecal egg count reduction were performed and yielded different results. A new method based on the estimated faecal egg counts (corrected from the initial faecal egg counts and animal type using a general linear model) was used and gave higher flexibility in the interpretation of putative resistance to anthelmintics. From necropsy results, it could be concluded that Cooperia oncophora was resistant to avermectins and benzimidazoles and that Cooperia punctata, Ostertagia ostertagi and Haemonchus placei were resistant to benzimidazoles. This case of multispecies and multidrug resistance is probably not unique and could reflect the emergence of resistances in Argentina. This emergence is probably due to the intensive use of anthelmintics, the absence of refugia, and the frequent circulation of infected cattle.     

Relation of Japanese <Emphasis Type="Italic">Xanthomonas campestris</Emphasis> pv. <Emphasis Type="Italic">vesicatoria</Emphasis> with worldwide strains revealed with three specific monoclonal antibodies

Strains of Xanthomonas campestris pv. vesicatoria Dye 1978 (Xcv), the causal agent of bacterial spot, have been classified into two groups based on their ability to hydrolyze starch. Three monoclonal antibodies (MAbs), 7AH10, 5HB3, and 4AD2, were produced immunized against the living bacteria and were specific to and could distinguish Xcv strains able or unable to hydrolyze starch (Amy⁺ or Amy^–). The MAb 7AH10, obtained against strain UPB141(Amy^–) reacted in an enzyme-linked immunosorbent assay with all the Amy^– strains (n = 19) and 1 of 11 Amy⁺ strains. Against Xcv 2625, an Amy^– unusual phenotype strain, MAb 5HB3, recognized 97% of our worldwide collection of Xcvs (n = 30). Also against that strain, the MAb 4AD2 reacted with none of the homologous Amy^– phenotypes and with 90% (n = 11) of the heterologous Amy⁺ phenotypes. For all the MAbs, cross reactions with other pathovars or species were less than 4% (n = 67). By assaying a Japanese collection of strains against the three MAbs, the Amy⁺ strains were distinguished from the Amy^– strains, and their relation with other world strains could be demonstrated. All the MAbs reacted with the lipopolysaccharide fraction of the bacterial cell wall during immunoblotting.     

Evaluation of a specific immunochemotherapy for the treatment of canine visceral leishmaniasis

The efficacy of specific immunochemotherapy against Leishmania infantum infection in dog was studied. The effects on transmission of the disease, as well as the cellular and humoral immune response were examined. The treated animals showed a significant reduction in the infection rates that were detected in Phlebotomus perniciosus females fed on the dog. The humoral immune response, assayed with an indirect immunofluorescence antibody test (IFAT), did not show significant variations under the influence of the therapy. The characterisation of the peripheral blood mononuclear cells (PBMC) using flow cytometry indicated a significant increase in the proportion of T lymphocytes, especially of CD4/TcR(alpha)(beta)(+) and CD4/CD45RA(+) cells, without showing evidence for modifications in the other leukocyte subsets. Cellular lymphoproliferation studies indicated a lack of a specific response to soluble leishmanial antigen (SLA), but the non-specific lymphoproliferative capacity assayed with phytohemagglutinin (PHA) was maintained.     

Goat paratuberculosis in Chile: first isolation and confirmation of Mycobacterium avium subspecies paratuberculosis infection in a dairy goat.

In October 2004, 41 goats > 2 years old from a Saanen dairy goat herd located in Purranque County, 10th Region, Chile, were sampled and tested for paratuberculosis. While collecting samples it was observed that several goats were thin and emaciated. One goat was sufficiently debilitated to warrant humane euthanasia. This animal was brought to the Veterinary School at the Universidad Austral de Chile for necropsy. The goat selected for necropsy was a 12-year-old doe. The animal showed classical clinical signs of caprine paratuberculosis: emaciation despite willingness to eat, dry and rough hair coat, and no evidence of diarrhea. Gross pathology and histopathology of the necropsied goat were consistent with paucibacillary paratuberculosis. Bacteriology, serology, and PCR confirmed the diagnosis. This is the first published report of goat paratuberculosis in Chile confirming a case of caprine paucibacillary paratuberculosis.     

Diagnosis of paratuberculosis by fecal culture and ELISA on milk and serum samples in two types of Chilean dairy goat herds.

Fecal culture has been the primary method used to diagnose paratuberculosis in goats. It is laborious, slow, and expensive. Validation of enzyme-linked immunosorbent assays (ELISAs) on milk samples could make paratuberculosis testing more widely available for goat farmers. The aim of this study was to determine the accuracy of serum and milk ELISAs for paratuberculosis, relative to fecal culture, in Chilean dairy goats. Eight dairy goat herds were selected. Feces, blood, and milk samples were collected from all female goats >2 years old. Fecal samples were cultured using Herrold egg yolk medium with mycobactin J and antibiotics. Serum and milk samples were tested using a commercial ELISA kit for Mycobacterium avium subsp. paratuberculosis antibody detection. A total of 383 goats were tested by ELISA and fecal culture. The sensitivity of ELISA on serum and milk relative to fecal culture was 74.3% (95% CI: 59.8-88.8) and 60% (95% CI: 43.8-76.2), respectively. The corresponding values for ELISA specificity based on the percentage of non- M. avium subsp. paratuberculosis-infected goats testing ELISA-negative were 98.6% (95% CI: 96.6-100) and 99.3% (95% CI: 97.9-100) on serum and milk, respectively. Proportions of positive results for serum and fecal samples were significantly different, whereas the proportions of positive results for milk and fecal samples were not significantly different. The milk ELISA had a moderate level of agreement with fecal culture results (Kappa = 0.57). The paratuberculosis ELISA on goat milk samples may be a cost-effective, accurate alternative to fecal culture.     

Comparative study of the characteristics of the carcass and the meat of the Chato Murciano pig and its cross with Iberian pig, reared indoors

The Chato Murciano pig is a breed catalogued as a Special Protection breed in danger of extinction. At present it is commonly reared in an intensive system. We have studied the mineral composition and other characteristics of its carcass and meat. Samples from the Musculus longissimus lumborum were taken from a total number of 43 animals, divided into two groups, Chato Murciano and Chato Murciano crossed with Iberian, reared in a conventional indoor system. The resulting meat of the Chato Murciano pig and its cross with Iberian was especially rich in iron and copper, rich in phosphorus, and marginally poor in calcium and sodium. The evolution of the pH of the carcass was that of pigs which do not produce PSE (pale, soft and exudative) meat. Some relevant positive correlation coefficients between minerals (P ≤ 0.05) have been found.