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.     

Enzyme activities and diuron persistence in soil amended with vermicompost derived from spent grape marc and treated with urea

Mineral fertilizers, organic amendments, and pesticides are inputs commonly used in conventional farming practices. The aim of this study was to evaluate the effects of single or combined applications of spent grape marc-vermicompost, urea, and/or diuron on soil-enzyme activities and the persistence of this herbicide in soils with low organic carbon content. The application of vermicompost enhanced dehydrogenase (DHase) enzyme activity over time but altered soil urease activity to a very limited extent. The reduction in diuron concentrations and the increase in DHase activity indicated that the soil microorganisms were capable of degrading the ureic herbicide. Treatment with vermicompost and diuron had a stimulatory effect on soil microbial activity. On the whole, the application of diuron and urea to the vermicompost-amended soil raised DHase and urease activity to maximum levels (>3 μg INTF g^?1 h^?1 and >47 μg NH₄⁺ g^?1 h^?1, respectively). The application of urea to the unamended and vermicompost-amended soil decreased diuron persistence from 18.8 and 33 d to 12.5 and 15 d, respectively. Our findings show that although vermicompost additions reduce diuron availability, this boosts diuron degradation when combined with urea. These additions, under different soil management conditions, minimize the bioavailability and persistence of diuron and consequently the risk of leaching and seepage into aquifers. Compared with untreated soils, these types of treated soils could also improve agricultural sustainability and the quality of the environment.     

Long term improvement in the treatment of canine leishmaniosis using an antimony liposomal formulation

Pharmacokinetic and clinical effectiveness of liposome-encapsulated N-methylglucamine antimoniate (LMA) was performed in dogs suffering from experimental leishmaniosis. LMA was compared with N-methylglucamine antimoniate (MGA), the same drug in its free form. Sb plasma concentrations for LMA were always higher than those for MGA. Mean residence time (MRT), half-life time (t(1/2)) and clearance (Cl) showed that Sb was eliminated slower after liposome administration. The high volume of distribution (Vd) obtained with LMA suggests that Sb could achieve therapeutic concentrations in parasite-infected tissues. Average plasma concentration at steady state (Css(ave)) shows that Sb body concentrations after LMA treatment (9.8 mg/kg Sb, each 24h) would be effective in Leishmania infantum canine infection. Comparing LMA with MGA in a 1-year follow-up we observed no relapses for LMA and total protein and gammaglobulin concentrations were within normal range, while for MGA both began to rise 3 months after treatment. Use of antimonial liposomal formulations may restore effectiveness to an existing drug and reduce toxicity.     

Nested PCR for diagnosis of canine leishmaniosis in peripheral blood, lymph node and bone marrow aspirates

A nested polymerase chain reaction (PCR) assay was developed using primers selected from the genomic DNA of Leishmania infantum and applied to the diagnosis of leishmaniosis in peripheral blood in dogs. Blood of 39 dogs of different breeds, all sampled in Catalonia (Spain), were tested for leishmaniosis by enzyme linked immunosorbent assay (ELISA), western blotting (WB) and peripheral blood mononuclear cell (PBMC) culture and nested PCR. Twenty negative controls (healthy dogs less than 1-year-old that had not been exposed to a sandfly season) were also studied. Nineteen of the 39 dogs studied were positive by ELISA and/or WB, and 18 of these had a positive PBMC nested PCR. PBMC nested PCR was negative in all the remaining animals that were negative by serological examination, including the 20 negative controls. Parasitological examination and nested PCR of bone marrow and lymph node aspirate from the 19 dogs positive by serological examination, were also positive. These results indicate that PBMC nested PCR is a sensitive and specific tool to diagnose leishmaniosis in dogs. The use of PBMC has the advantage over bone marrow and lymph node aspirates in that it is a less invasive sample.     

Evaluation of Natural Chemical Compounds Against Root-lesion and Root-knot Nematodes and Side-effects on the Infectivity of Arbuscular Mycorrhizal Fungi

The survival of two species of plant parasitic nematodes: the root-lesion nematode Pratylenchus brachyurus, and the root-knot nematode Meloidogyne javanica, was evaluated in saturated atmospheres of 12 natural chemical compounds. The infectivity of two isolates of arbuscular mycorrhizal fungi: Glomus mosseae and Glomus intraradices, under identical experimental conditions, was also determined. All the compounds tested exerted a highly significant control against M. javanica and among them, benzaldehyde, salicilaldehyde, borneol, p-anisaldehyde and cinnamaldehyde caused a mortality rate above 50% over P. brachyurus. The infectivity of G. intraradices was inhibited by cinnamaldehyde, salicilaldehyde, thymol, carvacrol, p-anisaldehyde, and benzaldehyde, while only cinnamaldehyde and thymol significantly inhibited mycorrhizal colonization by G. mosseae.     

Coinfection by Cryptosporidium parvum and porcine circovirus type 2 in weaned pigs

Routine histopathological diagnosis of one representative 3-month-old pig from a group suffering from diarrhoea revealed a massive degree of parasitation by Cryptosporidium parvum, with a concomitant infection by porcine circovirus type 2 (PCV2), that was confirmed by immunohistochemical procedures. The areas of intestine where parasites were more numerous presented abundant PCV2 infected cells in mucosa and submucosa. The concurrence of C. parvum, a rare primary intestinal pathogen in post-weaning and growing pigs, and PCV2 infections suggest an increased susceptibility as a result of an immunosuppression state.     

Development of Pleospora allii on Garlic Debris Infected by Stemphylium vesicarium

Pseudothecia of Pleospora allii developed best on garlic leaf debris infected by Stemphylium vesicarium incubated at low temperature (5–10°C) and relative humidity (RH) close to saturation. RH of less than 96% prevented the formation of pseudothecia, while an incubation temperature of 15–20°C led to the early degeneration of pseudothecia. Under natural conditions, colonization by pseudothecia of unburied garlic leaf debris varied between seasons from 6.0 to 15.5 pseudothecia/mm2, whereas lower colonization levels were recorded when samples were buried. Pseudothecial maturity was reached 1–4mo after the deposition of garlic debris on the soil surface and 15 days after the burial of residues. In the later case, pseudothecia degenerated with degradation of the plant debris. Ascospore release, which required rainfall or dew periods, occurred between late January and late April depending upon the year. A high correlation was found between pseudothecia maturation and four meteorological variables. Two of which, i.e. the number of hours with RH98% and with a mean temperature of 4.5–10.5°C, and the accumulated rainfall, explained most variability (adjusted R2=0.82–0.98 depending upon the year). A multiple regression equation relating the pseudothecia maturity index with these two variables could be used to forecast the epidemic onset of Stemphylium leaf spots in Southern Spain. Temporal progress of pseudothecia maturation was best fitted by a monomolecular model.