Leishmania spp. in Didelphis albiventris and Micoureus paraguayanus (Didelphimorphia: Didelphidae) of Brazil

Leishmaniasis is kept in nature by the participation of several animal species. This study evaluated the presence of Leishmania spp. in skin samples of free-ranging marsupials Micoureus paraguayanus (n=95) and Didelphis albiventris (n=191), captured in Morro do Diabo State Park and in sections of its surrounding forest, in the region of Pontal do Paranapanema, S?o Paulo State, Brazil. The samples were tested for the presence of kDNA of Leishmania spp. by polymerase chain reaction (PCR) and by real time PCR (qPCR). All samples from D. albiventris tested by PCR were negative for the presence of kDNA of Leishmania spp. However, when tested by qPCR, the positivity was 1.6%. A positivity of 7.4% by PCR and 11.6% by qPCR was observed for M. paraguayanus. Sixty-four per cent (9/14) of positive animals were limited to the same forest fragment. Presence of Leishmania (Leishmania) amazonensis and Leishmania (Viannia) braziliensis was detected in M. paraguayanus samples. While D. albiventris is the most studied marsupial species due to its urban habits, other marsupial species such as M. paraguayanus can be potential reservoirs of Leishmania spp. and should also be studied.     

Population genetics of Toxoplasma gondii: new perspectives from parasite genotypes in wildlife

Toxoplasma gondii, a zoonotic protozoal parasite, is well-known for its global distribution and its ability to infect virtually all warm-blooded vertebrates. Nonetheless, attempts to describe the population structure of T. gondii have been primarily limited to samples isolated from humans and domesticated animals. More recent studies, however, have made efforts to characterize T. gondii isolates from a wider range of host species and geographic locales. These findings have dramatically changed our perception of the extent of genetic diversity in T. gondii and the relative roles of sexual recombination and clonal propagation in the parasite's lifecycle. In particular, identification of novel, disease-causing T. gondii strains in wildlife has raised concerns from both a conservation and public health perspective as to whether distinct domestic and sylvatic parasite gene pools exist. If so, overlap of these cycles may represent regions of high probability of disease emergence. Here, we attempt to answer these key questions by reviewing recent studies of T. gondii infections in wildlife, highlighting those which have advanced our understanding of the genetic diversity and population biology of this important zoonotic pathogen.     

Evidence that the 36 kb plasmid of Brachyspira hyodysenteriae contributes to virulence

Swine dysentery (SD) results from infection of the porcine large intestine with the anaerobic intestinal spirochaete Brachyspira hyodysenteriae. Recently the genome of virulent Australian B. hyodysenteriae strain WA1 was sequenced, and a 36 kilobase (kb) circular plasmid was identified. The plasmid contained 31 genes including six rfb genes that were predicted to be involved with rhamnose biosynthesis, and others associated with glycosylation. In the current study a set of PCRs was developed to amplify portions of nine of the plasmid genes. When used with DNA extracted from virulent strain B204, PCR products were generated, but no products were generated with DNA from avirulent strain A1. Analysis of the DNA using pulsed field gel electrophoresis (PFGE) identified a plasmid band in strains WA1 and B204, but not in strain A1. These results demonstrate that strain A1 does not contain the plasmid, and suggests that lack of the plasmid may explain why this strain is avirulent. To determine how commonly strains lacking plasmids occur, DNA was extracted from 264 Australian field isolates of B. hyodysenteriae and subjected to PCRs for three of the plasmid genes. Only one isolate (WA400) that lacked the plasmid was identified, and this absence was confirmed by PFGE analysis of DNA from the isolate and further PCR testing. To assess its virulence, 24 pigs were experimentally challenged with cultures of WA400, and 12 control pigs were challenged with virulent strain WA1 under the same conditions. Significantly fewer (P=0.03) of the pigs challenged with WA400 became colonised and developed SD (13/24; 54%) compared to the pigs infected with WA1 (11/12; 92%). Gross lesions in the pigs colonised with WA400 tended to be less extensive than those in pigs colonised with WA1, although there were no obvious differences at the microscopic level. The results support the likelihood that plasmid-encoded genes of B. hyodysenteriae are involved in colonisation and/or disease expression.     

Immunization of chickens with Salmonella enterica subspecies enterica serovar Enteritidis pathogenicity island-2 proteins

Several structural components of the type III secretion systems (T3SS) encoded by Salmonella pathogenicity island (SPI)-1 and SPI-2 are exposed to the host's immune system prior to/during the infection/invasion process, making them potential vaccine candidates. In this study we evaluated whether chickens vaccinated with SPI-2 T3SS components could mount a significant humoral immune response (as measured by serum IgG titres) and whether these antibodies could be transferred to progeny (as measured by egg yolk IgG titres), and whether vaccinates and progeny of vaccinates could be protected against challenge with SE. The results of our studies show that vaccinated chickens do produce high levels of SPI-2 T3SS specific serum IgG that they are able to transfer to their progeny. It was demonstrated that vaccinates and progeny of vaccinates had lower overall countable recovered Salmonella enterica subspecies enterica serovar Enteritidis (SE) per bird in most situations.     

Interlaboratory evaluation of a real-time multiplex polymerase chain reaction method for identification of salmon and trout species in commercial products

This interlaboratory study evaluated a real-time multiplex polymerase chain reaction (PCR) method for identification of salmon and trout species in a range of commercial products in North America. Eighty salmon and trout products were tested with this method by three independent laboratories. Samples were collected in the United States and Canada, and only the collecting institution was aware of the species declaration. Following analysis with real-time PCR, all three laboratories were able to identify species in 79 of the 80 products, with 100% agreement on species assignment. A low level of fraud was detected, with only four products (5%) found to be substituted or mixtures of two species. The results for two of the fraudulent products were confirmed with alternate methods, but the other two products were heavily processed and could not be verified with methods other than real-time PCR. Overall, the results of this study show the usefulness and versatility of this real-time PCR method for the identification of commercial salmon and trout species.     

MONOPOTASSIUM PHOSPHATE AS AN IN-SEASON FERTIGATION OPTION FOR POTATO

Monopotassium phosphate (MKP) is a potential option for fertigating phosphorus (P) in potato (Solanum tuberosum L.) when petioles are low in P and high in nitrogen (N); which is a situation where using ammonium polyphosphate (APP) could potentially result in excessive N application. Fertilizer trials were conducted in 2004–2006 with 0 or 56 kg P₂O₅ ha^?1 fertigated as either APP or MKP as a supplement to the pre-plant P (112 or 224 kg P₂O₅ ha^?1) broadcast applied to all plots. Supplemental P fertigation increased petiole P concentration, US No. 1 yield, and total yield over the control not receiving any in-season P fertilizer regardless of source. In addition, MKP increased tuber specific gravity. These results support previous studies showing that fertigated P can be used to increase potato yields when petiole P concentrations are low and that MKP is a viable substitute for APP fertilizer when fertigation is necessary.     

Rapid impact of Impatiens glandulifera control on above‐ and belowground invertebrate communities

The annual plant Impatiens glandulifera (Himalayan balsam) is the most widespread invasive non‐native weed in the British Isles. Manual control is widely used, but is costly and laborious. Recently, biological control using the rust fungus Puccinia komarovii var. glanduliferae has been trialled. We designed an experiment to assess the impact of these control methods on invertebrate communities in relation to unmanaged and uninvaded habitats, and to determine whether mycorrhizal inoculation aided post‐control recovery of these communities. Sixty invaded and twenty uninvaded field soil blocks were transplanted to the experiment site, where a mycorrhizal inoculum was added to half of all blocks. Biological and mechanical control treatments were applied to twenty invaded blocks independently; the twenty remaining invaded blocks were left intact. Above‐ and belowground invertebrate samples were collected from the blocks at the end of the growing season. Overall, aboveground invertebrate abundance increased with the removal of I. glandulifera, and several groups showed signs of recovery within one growing season. The effect of mechanical control was more variable in belowground invertebrates. Biological control did not affect aboveground invertebrate abundance but resulted in large increases in populations of belowground Collembola. Our experiment demonstrates that mechanical removal of I. glandulifera can cause rapid increases in invertebrate abundance and that its biological control with P. komarovii var. glanduliferae also has the potential to benefit native invertebrate communities.     

Neoamphimedine circumvents metnase-enhanced DNA topoisomerase IIα activity through ATP-competitive inhibition

Type IIα DNA topoisomerase (TopoIIα) is among the most important clinical drug targets for the treatment of cancer. Recently, the DNA repair protein Metnase was shown to enhance TopoIIα activity and increase resistance to TopoIIα poisons. Using in vitro DNA decatenation assays we show that neoamphimedine potently inhibits TopoIIα-dependent DNA decatenation in the presence of Metnase. Cell proliferation assays demonstrate that neoamphimedine can inhibit Metnase-enhanced cell growth with an IC(50) of 0.5 μM. Additionally, we find that the apparent K(m) of TopoIIα for ATP increases linearly with higher concentrations of neoamphimedine, indicating ATP-competitive inhibition, which is substantiated by molecular modeling. These findings support the continued development of neoamphimedine as an anticancer agent, particularly in solid tumors that over-express Metnase.     

Competitive interactions between juvenile trembling aspen and lodgepole pine: A comparison of two interior British Columbia ecosystems

To facilitate ecosystem-specific management of juvenile mixtures of lodgepole pine (Pinus contorta Dougl. Ex Loud. Var. latifolia Engelm.) and trembling aspen (Populus tremuloides Michx.) in south-central British Columbia, we compared the characteristics of pine–aspen competition between a moist sub-boreal spruce and a dry interior Douglas-fir ecosystem. A total of 252 lodgepole pine and their neighbourhoods were examined across four untreated stands, each of which was sampled three times between ages 12 and 24 years. Pine diameter and height decreased with increasing density of trembling aspen at least as tall as the target pine (tall aspen) in both ecosystems. Regression analysis was used to examine the ability of tall aspen density and four competition indices (CIs) to predict pine size. Tall aspen density, which is easily assessed in the field, accounted for 63% and 69% of the variation in pine diameter and height in 20–24 year-old stands, respectively. The most successful competition index, based on the basal diameter ratio (BDR) of trembling aspen to pine accounted for, respectively, 78% and 73% of the variation. In the same stands, R² values were 1–5% lower when tall aspen density and BDR at age 15–19 years were used to predict size of 20–24-year-old pine.