Development of an indirect ELISA-NcSRS2 for detection of Neospora caninum antibodies in cattle

Neosporosis is of alarming economic concern in the cattle industry. The effectiveness of diagnostic tests for detecting specific antibodies against Neospora caninum is hampered by potential cross-reaction with other coccidia. Use of a single specific antigen might improve test specificity. An indirect enzyme-linked immunosorbent assay (ELISA) was developed using the truncated protein NcSRS2 expressed in Escherichia coli. The ELISA results were compared with those of the indirect fluorescence antibody test (IFAT). Receiver Operating Characteristic (ROC) and Tests in the Absence of a Gold Standard (TAGS) analysis revealed an assay having 96% specificity and 95% sensitivity when applied to 145 positive and 352 negative sera from two distinct cattle populations. Using OD ≤ 0.095 as the cut-off point, the assay's negative and positive predictive values ranged from 98.8% to 50.8% and from 58.8% to 99.1%, respectively, depending on neosporosis prevalence in a given area. The novel ELISA-NcSRS2 format described in the present report constitutes a specific and sensitive method for detecting N. caninum in cattle.     

Larval development of Toxocara canis in dogs

The parasitic roundworm Toxocara canis is present in dog populations all over the world. Due to its zoonotic potential, this roundworm is of special interest not only for veterinarians, but also for medical practitioners. In the present review, current knowledge of infection routes and the subsequent development of larvae within the canine host is summarised. Furthermore, information about the clinical, pathological, enzymatic, haematological and histopathological changes was collected, giving a broad overview of current knowledge of the infection. Although the data collected over the years give an idea of what happens during the larval development of T. canis, many questions remain open. Nevertheless, it is important that we continue our efforts to further understand the biology of this versatile and compelling parasite and try to improve and optimise strategies to prevent the infection in dogs and thereby to protect humans from this infection.     

Acute toxoplasmosis in squirrel monkeys (Saimiri sciureus) in Mexico

Toxoplasma gondii causes fatal multisystemic disease in New World primates, with respiratory failure and multifocal necrotic lesions. Although cases and outbreaks of toxoplasmosis have been described, there are few genotyping studies and none has included parasite load quantification. In this article, we describe two cases of lethal acute toxoplasmosis in squirrel monkeys (Saimiri sciureus) of Mexico city. The main pathological findings included pulmonary edema, interstitial pneumonia, hepatitis and necrotizing lymphadenitis, and structures similar to T. gondii tachyzoites observed by histopathology in these organs. Diagnosis was confirmed by immunohistochemistry, transmission electron microscopy and both end point and real time PCR. The load was between <14 and 23 parasites/mg tissue. Digestion of the SAG3 gene amplicon showed similar bands to type I reference strains. These are the first cases of toxoplasmosis in primates studied in Mexico, with clinical features similar to others reported in Israel and French Guiana, although apparently caused by a different T. gondii variant.     

Isolation and characterisation of equine influenza viruses (H3N8) from Europe and North America from 2008 to 2009

Like other influenza A viruses, equine influenza virus undergoes antigenic drift. It is therefore essential that surveillance is carried out to ensure that recommended strains for inclusion in vaccines are kept up to date. Here we report antigenic and genetic characterisation carried out on equine influenza virus strains isolated in North America and Europe over a 2-year period from 2008 to 2009. Nasopharyngeal swabs were taken from equines showing acute clinical signs and submitted to diagnostic laboratories for testing and virus isolation in eggs. The sequence of the HA1 portion of the viral haemagglutinin was determined for each strain. Where possible, sequence was determined directly from swab material as well as from virus isolated in eggs. In Europe, 20 viruses were isolated from 15 sporadic outbreaks and 5 viruses were isolated from North America. All of the European and North American viruses were characterised as members of the Florida sublineage, with similarity to A/eq/Lincolnshire/1/07 (clade 1) or A/eq/Richmond/1/07 (clade 2). Antigenic characterisation by haemagglutination inhibition assay indicated that the two clades could be readily distinguished and there were also at least seven amino acid differences between them. The selection of vaccine strains for 2010 by the expert surveillance panel have taken these differences into account and it is now recommended that representatives of both Florida clade 1 and clade 2 are included in vaccines.     

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.     

Gene flow among different teosinte taxa and into the domesticated maize gene pool

Maize (Zea mays L. ssp. mays) was domesticated from one wild species ancestor, the Balsas teosinte (Zea mays ssp. parviglumis) about 9000 years ago. Higher levels of gene diversity are found in teosinte taxa compared to maize, following domestication and selection bottlenecks. Diversity in maize can be increased via gene flow from teosinte, which has certainly occurred from various taxa, but the rate of flow from different teosinte taxa and the final impact on maize evolution has been difficult to measure. One hundred populations from six Zea taxa, both domesticated (maize) and wild (teosinte), including domesticated landraces from Asia, Africa, and the Americas, were genotyped with 17 SSR markers using 15 individuals per population. Overall levels of diversity were high, and populations could be distinguished based on markers. Relationships between populations followed most published reports, or can now help resolve previously conflicting reports. Gene flow into maize from different teosinte groups, and gene flow between different teosintes, was estimated. Evidence for contributions from the Balsas teosintes and from Chalco teosintes (Z. mays ssp. mexicana) to the maize gene pool was found, as well as from Chalco into ssp. mexicana race ??Durango?? and Z. mays ssp. huehuetenengensis. These contributions are almost certainly the result of post-domestication (and ongoing) exchanges. This information must give more impetus to in situ conservation of teosinte species, and use of these teosintes to continue to direct the evolution of maize, especially in response to new diseases, insect pests, and other biotic and abiotic stresses.     

Lindane Biodegradation by Defined Consortia of Indigenous Streptomyces Strains

The current study aimed to compare lindane degradation by pure and mixed cultures of Streptomyces sp. Cell-free extracts were assayed for potentiating dechlorinase activity and, based on these results, consortia of two to six microorganisms were assayed for their growth on and degradation of lindane. Furthermore, the role of bacterial consortia of lindane-degrading strains was examined in lindane decontamination soil assays. Four actinobacteria, previously isolated from a pesticide-contaminated area, were selected because of their tolerance to lindane and their ability to use the pesticide as sole carbon source. These strains as well as Streptomyces sp. M7 and Streptomyces coelicolor A3 were used to study specific dechlorinase activity (SDA) and lindane removal in mixed cultures. Pure cultures presented SDA in the presence of 1.66 mg L^-1 lindane as carbon source. SDA was improved by certain mixed cultures until 12 times compared with pure cultures. Mixed cultures with two, three, and four strains showed maximum lindane removal of 46% to 68%, whereas combinations of five and six strains did not efficiently remove the pesticide from the culture medium. The Streptomyces sp. A2, A5, M7, and A11 consortium presented the lowest ratio between residual lindane concentration and SDA and could be a promising tool for lindane biodegradation.     

Applying a Lagrangian dispersion analysis to infer carbon dioxide and latent heat fluxes in a corn canopy

Warland and Thurtell (2000) proposed an analytical dispersion Lagrangian analysis (hereafter WT analysis) to relate the mean scalar concentration field to source profiles inside the canopy. The first objective of this study was to evaluate the performance of the WT analysis with existing turbulence statistics parameterizations in a corn canopy, by comparing its inferred net ecosystem CO₂ exchange (NEE) and latent heat flux (λE) with eddy covariance measurements. The second objective was to assess the performance of the WT analysis to infer the soil CO₂ flux. Four parameterizations of turbulence statistics were used to estimate Lagrangian time scale (T_L) and standard deviation of vertical wind velocity (σ_w) profiles. The estimated T_L and σ_w profiles were then corrected for atmospheric stability conditions. The field experiment was carried out in a corn field from August to October 2007 and 2008. Profiles of water vapour and CO₂ mixing ratios were measured using a multiport sampling system connected to an infrared gas analyzer. Wind velocity within and above the canopy and eddy covariance measurements over the canopy were taken. The soil respiration, estimated using the WT analysis, was compared to estimates obtained by an empirical model. WT analysis fluxes showed good correlation (R² = 0.77-0.88) with NEE and λE obtained by the eddy covariance technique, but overestimated net fluxes, especially when corrections for atmospheric stability were applied. The optimization of T_L and σ_w profiles using in-canopy turbulence measurements improved the agreement between measured and modeled NEE and λE. Inferred soil CO₂ fluxes were underestimated and were poorly correlated (R² = 0.02-0.01) with estimates obtained using an empirical model based on soil temperature. This poor performance in estimating the soil respiration is likely caused by the decoupling between inside and above canopy flows.