Experimental evidence of indirect transmission of Mycoplasma synoviae

The aim of the study was to analyse experimental transmission of Mycoplasma synoviae, an avian pathogen. Three experiments using specific pathogen-free day-old chicks placed in isolators were conducted. In the first experiment, the birds were introduced in an isolator previously contaminated with a M. synoviae broth culture. After 34 days, these birds were eliminated and, for the second trial, the chicks were introduced in the same isolator without disinfecting. In the third assay, the chicks were placed in an isolator containing a mixture of food, feathers and dust collected less than an hour earlier from a M. synoviae infected laying hen flock. In the second and third experiments in order to exacerbate the M. synoviae infection, the birds were inoculated with infectious bronchitis (IB) virus. The presence of M. synoviae in the environment and in tracheal swabs was monitored by culture, a multiplex PCR (mPCR) detecting M. synoviae and Mycoplasma 16S rDNA and a multiplex RT-PCR (mRT-PCR) detecting the M. synoviae mRNA coding for a membrane protein and Mycoplasma 16S rRNA. In in vitro experimental conditions, M. synoviae mRNA and 16S rRNA were detected up to 20 min and 23 h respectively after mycoplasma death. In the first assay, the first infected bird was detected on the 13th day. In the second trial, culturable M. synoviae or viable M. synoviae were detected in the isolator for 3 or 4 to 5 days respectively after depopulation of the birds of the first assay whereas the first culture positive tracheal swabs were detected on the 33rd day, after IB inoculation. In the third experiment, the first infected birds were detected on the 54th day. Thus, the different assays showed that M. synoviae contaminated material (dust, feathers and food) can infect chicks, sometimes after remarkably long silent periods.     

Time effect on bentazone sorption and degradation in soil

Previous sorption/desorption batch experiments have indicated that bentazone is weakly sorbed by soils. In addition, field experiments have shown that 4% of the bentazone sprayed can be leached to drainage water. In order to complete bentazone characterisation, we have assessed the effect of time on its behaviour in contrasting soils. In laboratory studies, bentazone was added to three topsoils (sandy, loamy and clay soils). Bentazone degradation, sorption/desorption kinetics and isotherm measurements were carried out at different times. At 160 days after treatment, bentazone mineralisation amounts varied from 2.1% (sandy soil) to 14% (clay soil). The extractable amounts became lower (from 97% after treatment to 12% after 160 days for the clay soil) and a greater number of desorption series was needed to obtain these products. Nevertheless, at the end of the experiments, a small amount of bentazone was still extracted by water. At the same time, bound residues of bentazone reached 65% in clay soil. Statistical analysis indicated effects of both residence time and soil type on bentazone behaviour.     

A recombinant 31.5 kDa keratinase and a crude exo-antigen from Microsporum canis fail to protect against a homologous experimental infection in guinea pigs

A Microsporum canis recombinant 31.5 kDa keratinase and a M. canis crude exo-antigen were tested as vaccines in an experimental infection model in guinea pigs. Animals were vaccinated subcutaneously three times at two-week intervals with either the keratinase, the exo-antigen or the adjuvant alone. Cutaneous challenge was performed blindly. Both humoral and cellular-specific immune responses to M. canis antigens were evaluated every 14 days, while a blind evaluation of clinical lesion development and fungal persistency in skin were monitored weekly. Vaccination induced very high and significant (P < 0.01) antibody responses towards both antigens. High cell-mediated immune responses to both immunogens were also induced by vaccination. After challenge, however, scores reflecting the severity of dermatophytic lesions did not differ significantly between vaccinated and control groups at any time after challenge. These results suggest that, in the guinea pig, the induction of specific immune responses against the M. canis-secreted antigens used in this study are not protective against challenge exposure.     

Wheat leaf rust uredospore production and carbon and nitrogen export in relation to lesion size and density

ABSTRACT To develop mechanistic yield loss models for biotrophic fungi, we need better account for the export of dry matter, carbon, and nitrogen from the leaf into the spores. Three experiments in controlled environment chambers were performed to study the dynamics of uredospores production of Puccinia triticina on seedling leaves of wheat in relation to time, lesion density, and sporulating surface area. The detrimental effect of lesion density on the sporulation capacity of brown rust lesions was confirmed. When lesion density increased, spores production per lesion strongly decreased. However, our results showed that increasing lesion density also greatly reduces lesion size. A model was developed to summarize these relationships. Our main conclusion is that the density effect on spore production per lesion is accounted for by lesion size. When sporulation was related to the sporulating surface area, it became independent of density. As well, carbon and nitrogen contents of the spores were independent of lesion density. Our data suggest that when nitrogen available in the host is limiting, spore production is reduced but nitrogen content of spores tend to remain stable.     

HAEMATOLOGICAL INVESTIGATION OF A MULTIPLE CASE LEUCOSIS HERD

Adult cattle in a Queensland dairy herd with a history of deaths from lymphosarcoma were sampled regularly over a 4 year period for the identification of animals with persistent lymphocytosis (PL). Twenty-one of 94 animals that were sampled at least 6 times had PL. At the initial sampling 27% of the animals had lymphocytosis. Culling of haematologically positive animals in the first 18 months of the investigation reduced this to 5.3%, but cessation of the culling programme resulted in a gradual increase in the percentage of animals with lymphocytosis. Four deaths from lymphosarcoma occurred in adult animals, but only in the first 18 months of the investigation. Two of these animals had lymphocytosis and two lymphoblastic leukaemia. The calf of one of the latter cows developed lymphoblastic leukaemia and lymphosarcoma by the time it was 6 months of age. Although histological evidence of lymphosarcoma was lacking in a number of clinically normal animals with lymphocytosis, haematological investigation identified a group of animals within the herd that may develop lymphosarcoma.     

Efficacy of hexaflumuron against the fungus-growing termite Pseudacanthotermes spiniger (Sjöstedt) (isoptera,macrotermitinae)

The efficacy of hexaflumuron, a benzophenylurea insecticide, has been studied for the first time on a fungus-growing termite (Pseudacanthotermes spiniger Sjöstedt, Macrotermitinae). Results show that hexaflumuron could be useful in treating infestations of such pest species, which are of great economic importance in many tropical and equatorial countries. Foraging workers harvested and introduced treated food into the nest and subsequently contaminated the brood by trophallaxis. Hexaflumuron showed potent larvicidal activity. The compound did not appear to be rapidly degraded by the digestive enzymes of termite workers, nor by the symbiotic fungus Termitomyces eurhizus Heim growing on fungus combs. This chitin synthesis inhibitor did not act as a fungicide, since growth of the mycelium of this fungus was not inhibited in vitro. © 1998 Society of Chemical Industry     

Identification of <Emphasis Type="Italic">Ditylenchus</Emphasis> species associated with Fabaceae seeds based on a specific polymerase chain reaction of ribosomal DNA-ITS regions

A technique based on the use of specific primers for polymerase chain reaction (PCR) was developed for the identification of the stem and bulb nematode belonging to the Ditylenchus dipsaci species complex. The internal transcribed spacer region ITS1 and ITS2, the gene 5.8 S and part of genes 18 S and 26 S of twenty populations of the D. dipsaci species complex belonging to both D. dipsaci sensu stricto and Ditylenchus sp. B (corresponding to populations of giant individuals associated to Vicia faba) and three congeneric species were amplified with two universal ribosomal primers. PCR-amplified DNA samples were digested with five restriction enzymes in order to reveal some polymorphism allowing the identification of D. dipsaci populations associated with Fabaceae seeds. The polymorphism among species was confirmed by the sequencing of the PCR products. A primer (DdpS2) was designed in a region conserved in all populations of both D. dipsaci sensu stricto and D. sp. B studied in the present work. The other Anguinidae species (except a few species from Central Asia associated to Astereaceae and D. sp. G associated to Plantago maritima) differ in two to four nucleotides at the 3′ extremity of this region. This sequence portion coincides with a TspEI restriction site. In combination with a primer located in the ribosomal region, this first primer is a good candidate for identification by PCR of populations of the D. dipsaci species complex found in Fabaceae seeds. A second primer (DdpS1) was designed in a similar way and was specific to D. dipsaci sensu stricto. The utility of these two sets of primers is discussed against the background of quarantine regulation.     

Molecular characterization of Streptococcus suis strains by 16S–23S intergenic spacer polymerase chain reaction and restriction fragment length polymorphism analysis

We developed a new molecular method of typing Streptococcus suis based on polymerase chain reaction (PCR) amplification of a large fragment of rRNA genes, including a part of the 16S and 23S genes and the 16S-23S intergenic spacer region (ISR), followed by restriction fragment length polymorphism (RFLP) analysis with RsaI or MboII endonuclease. The 16S-23S ISRs of 5 S. suis isolates were sequenced and compared. Size and sequence polymorphisms were observed between the S735 reference strain and the 4 wild-type strains. The genetic relationships between 138 independent S. suis strains belonging to various serotypes, isolated from swine or human cases, were determined. The discriminatory power of the method was > 0.95, the threshold value for interpreting typing results with confidence (0.954 with RsaI and 0.984 with RsaI plus MboII). The in vitro reproducibility was 100%. The strains isolated from humans were less genetically diverse than the strains isolated from pigs. For the first time, 2 molecular patterns (R6, M9) were significantly associated with S. suis serotype 2 strains. This genetic tool could be valuable in distinguishing individual isolates of S. suis during epidemiologic investigations.     

Sulcotrione versus atrazine transport and degradation in soil columns

A soil column experiment under outdoor conditions was performed to monitor the fate of 14C-ring-labelled sulcotrione, 2-(2-chloro-4-mesylbenzoyl)cyclohexane-1,3-dione and atrazine, 6-chloro-N2-ethyl-N4-isopropyl-1,3,5-triazine-2,4-diamine, in water leachates and in the ploughed horizon of a sandy loam soil. Two months after treatment, the cumulative amounts of herbicide residues leached from the soil were 14.5% and 7% of the applied radioactivity for sulcotrione and atrazine, respectively. Maximum leachate concentrations for each herbicide were observed during the first month following application: 120 and 95 microg litre(-1) for sulcotrione and atrazine respectively. After 2 weeks, 78% of the sulcotrione and atrazine was extractable from the soil, whereas after two months only 10 and 4%, respectively, could be extracted. The maximum sulcotrione content in the first 10 cm of soil was identical with that of atrazine. For both molecules, the content of non-extractable residues was low, being around 15%. Sulcotrione seems to be more mobile than atrazine but the consequences for water contamination are similar since lower doses are used.