Assessing the role of sow parity on PRRSv detection by RT-qPCR through weekly processing fluids monitoring in breeding herds.

The use of processing fluids to monitor the breeding herd's porcine reproductive and respiratory syndrome (PRRS) status has gained industry acceptance. However, little is known about PRRS virus RT-qPCR detection dynamics in processing fluids and factors that may contribute to maintain PRRS virus in the herd after an outbreak. This study aimed to describe weekly RT-qPCR processing fluid results in breeding herds after an outbreak and to evaluate the proportion of RT-qPCR positive results among parity groups. Processing tissues of 15 first parity (P1), 15 second parity (P2), and 15 third parity or higher (P3+) litters (parity groups) were collected weekly for between 19 and 46 weeks in nine breeding herds. Processing fluids were aggregated, and RT-qPCR tested by parity group weekly. Additionally, a subset of 743 processing fluid samples of litters that formed 50 parity groups, as previously described, were RT-qPCR tested individually at the litter level. The agreement between RT-qPCR results of processing fluid samples of parity groups (15 litters) and results based on individual litter testing was assessed using overall percent of agreement, Kappa statistic, and McNemar test. The association between RT-qPCR results and the parity group was evaluated using a generalized estimating equations model, after accounting for the effects of sampling week, breeding herd PRRS control strategy (i.e., open to replacements v/s closed) and herd. An autoregressive correlation structure was used to account for the repeated samplings within a herd in time. The overall agreement was 98 %, and Kappa statistic 0.955 (McNemar p = 1.0). Sensitivity of parity group processing fluid samples was estimated at 100 % (95 % CI 89-100 %), while specificity was estimated at 94 % (95 % CI 71-100 %). Although P1 aggregated litters had on average a higher proportion of RT-qPCR positive results from outbreak week 25 onwards, the proportion was not significantly different to the one observed for P2 and P3+ aggregated litters (p > 0.13). Additionally, herds that interrupted gilt entry had lower odds of PRRS RT-qPCR positivity than herds that continued entering gilts (OR = 0.35, 95 % CI 0.16-0.78). PRRS virus persistence in processing fluids was not affected by the sow parity effect in most of the breeding herds studied. No evidence of disagreement between RT-qPCR results of an aggregated sample of 15 litters and those of individual litters was observed. This level of litter aggregation testing strategy may be of particular use at the last stages of an elimination program under low PRRS virus prevalence.

Description of changes of key performance indicators and PRRSV shedding over time in a naïve breeding herd following a PRRS MLV exposure.

Porcine reproductive and respiratory syndrome (PRRS) is an important economic swine disease. The usage of PRRS-modified live vaccines (MLV) is the predominant breeding herd immunologic solution used in the United States to minimize the economic losses associated with wild-type PRRS infection. Most of the current information on the effects of contemporary PRRS MLV vaccination on breeding herd performance under field conditions comes from herds with previous PRRS virus (PRRSV) exposure. Hence, there is little information on key performance indicators (KPI) changes after the exposure to a PRRS MLV in PRRSV-naïve breeding herds. The main objective of this longitudinal observational study was to describe selected KPI changes in a naïve breeding herd after PRRS MLV exposure. The secondary objective was to describe the pattern of detection of PRRSV RNA by the quantitative reverse transcriptase-polymerase chain reaction in processing fluid samples. There were transient increases for mummies during weeks 4-23 (+0.86%); increased pre-weaning mortality on weeks 3-5 (+3.76%); a decrease in live born on weeks 4-5 (-0.46) leading to a decreased pig weaned/litter on weeks 5-10 (-0.69) and increased repeated services on weeks 3-23 (+5.53%). Transient changes observed after PRRS MLV exposures did not move total pigs weaned to outside the control intervals. Starting on week 83 and for 53 consecutive weeks, there was no PRRSV detection in processing fluids, even though two whole-herd MLV exposures occurred within that period.

Reduction of Sb(V) by coupled biotic-abiotic processes under sulfidogenic conditions.

Increasing use and mining of antimony (Sb) has resulted in greater concern involving its fate and transport in the environment. Antimony(V) and (III) are the two most environmentally relevant oxidation states, but little is known about the redox transitions between the two in natural systems. To better understand the behavior of antimony in anoxic environments, the redox transformations of Sb(V) were studied in biotic and abiotic reactors. The biotic reactors contained Sb(V) (2 mM as KSb(OH)6), ferrihydrite (50 mM Fe(III)), sulfate (10 mM), and lactate (10 mM), that were inoculated with sediment from a wetland. In the abiotic reactors, The interaction of Sb(V) with green rust, magnetite, siderite, vivianite or mackinawite was examined under abiotic conditions. Changes in the concentrations of Sb, Fe(II), sulfate, and lactate, as well as the microbial community composition were monitored over time. Lactate was rapidly fermented to acetate and propionate in the bioreactors, with the latter serving as the primary electron donor for dissimilatory sulfate reduction (DSR). The reduction of ferrihydrite was primarily abiotic, being driven by biogenic sulfide. Sb and Fe K-edge X-ray absorption near edge structure (XANES) analysis showed reduction of Sb(V) to Sb(III) within 4 weeks, concurrent with DSR and the formation of FeS. Sb K-edge extended X-ray absorption fine structure (EXAFS) spectroscopy analysis indicated that the reduced phase was a mixture of S- and O-coordinated Sb(III). Reduction of Sb(V) was not observed in the presence of magnetite, siderite, or green rust, and limited reduction occurred with vivianite. However, reduction of Sb(V) to amorphous Sb(III) sulfide occurred with mackinawite. These results are consistent with abiotic reduction of Sb(V) by biogenic sulfide and reveal a substantial influence of Fe oxides on the speciation of Sb(III), which illustrates the tight coupling of Sb speciation with the biogeochemical cycling of S and Fe.

Nano-enabled, antimicrobial toothbrushes - How physical and chemical properties relate to antibacterial capabilities.

Over the past two decades, Ag and Zn nanoparticles have been integrated into various consumer products as a biocide. While some nano-enabled consumer products have been shown to have antibacterial properties, their antibacterial efficacy as well as the human and environmental health outcomes are not fully known. In this study, we examine a nanoparticle-enabled product that also serves as a conduit for human exposure to bacteria: toothbrushes. We utilize a combination of chemical analyses, laboratory experiments, and microscopy to characterize the nano-enabled toothbrush bristles. Our analysis showed the majority of measured Ag and Zn particles ranged from approximately 50 to 100 nm in size and were located on the surface and within bristles. During simulated brushing, antimicrobial bristles released both Ag and Zn, the majority of which was released in particulate form. While our results demonstrate that antimicrobial bristles have enhanced bactericidal properties compared to control samples, we also show that the surface topography influences nanoparticle retention, microbial adhesion, and bactericidal activity. We thus conclude that Ag or Zn content alone is insufficient to predict antimicrobial properties, which are further governed by the bioavailability of Ag or Zn at the bristle surface.

Assessment of immediate production impact following attenuated PRRS type 2 virus vaccination in swine breeding herds.

BACKGROUND: To mitigate production impact of porcine reproductive and respiratory syndrome (PRRS) virus outbreaks, it has been common to preventively vaccinate swine breeding herds using PRRS modified live virus (MLV) vaccine. However, attenuated PRRS virus (PRRSv) may result negative impact on farm productivity. The objective of this study was to measure the immediate impact of PRRS type 2 MLV vaccine on breeding herd performance under field conditions. Eight PRRS-stable farms routinely mass vaccinating females with commercial PRRS MLV vaccines were enrolled on study. Vaccination dates were collected and weekly changes in abortions, neonatal losses, pre-weaning mortality, pigs weaned per sow, and wean-to-first-service interval were assessed for up to 6 weeks after each vaccination. A 6-week period prior to each vaccination was established as baseline. Statistical process control (SPC) analysis was conducted to detect significant productivity decreases after MLV interventions, on each farm, and a mixed regression model was used, at the aggregated data level, to assess the productivity change 6 weeks after PRRS MLV vaccinations, compared to baseline. RESULTS: Out of 65 herd-MLV vaccinations, SPC analysis detected increase on abortions 4 times (6.1%), on neonatal losses 7 times (10.7%), on pre-weaning mortality 2 times (3%), on wean-to-first-service interval 2 times (3%), and no change in total pigs weaned. On aggregated data analysis, there was no significant change in abortion rate, neonatal losses, number of pigs weaned per sow, and wean-to-first-service interval. However, there was an increase of 0.26% of pre-weaning mortality 2 weeks after vaccination compared to the baseline. CONCLUSIONS: Under study conditions, individual PRRS-stable sow farms had experienced transient, and numerically small changes in productivity following PRRS type 2 MLV vaccination. There was a small increase of pre-weaning mortality 2 weeks after vaccination, but no evidence of significant production impact at aggregated data analysis for abortion rate, neonatal losses, pigs weaned per sow and wean-to-first-service interval.

Testing the component additivity approach to surface complexation modeling using a novel cadmium-specific fluorescent probe technique.

Over the past 40 years, laboratory experiments involving single metal-single sorbent systems have been conducted in order to determine thermodynamic stability constants for metal-bacteria and metal-mineral surface complexes. The component additivity (CA) approach to surface complexation modeling (SCM) represents one method for using these experimentally-derived stability constants to predict the extent of metal adsorption in complex, multi-sorbent systems. However, quantitative tests of the CA approach are rare due to difficulties in determining the distribution of metals in complex multi-sorbent systems. In this study, we use a novel technique that couples the use of a cadmium(Cd)-specific fluorescent probe with confocal scanning laser microscopy to quantify Cd adsorption to bacteria in fully hydrated multi-sorbent samples that contain different ratios of Bacillus subtilis bacterial cells, the clay mineral kaolinite, and the aqueous chelating ligand EDTA. In this approach, we directly determine the distribution of Cd by measuring the total concentration of adsorbed Cd and the concentration of Cd that is adsorbed to bacterial cells, and by difference we calculate the concentration of Cd that is adsorbed to kaolinite. We compare these experimental measurements to the extent of Cd adsorption that is calculated using a CA approach to predict the distribution of Cd under our experimental conditions. In general, the CA predictions of the distribution of Cd between the aqueous phase and the two sorbents agree within uncertainties with the measured concentrations of Cd in each reservoir in both the EDTA-free and the EDTA-bearing experimental systems. This study demonstrates that the Cd-fluorescent probe technique is a suitable, and relatively simple, option for quantitatively testing CA surface complexation models. Our results suggest that although the CA approach can yield reasonable predictions of the distribution of Cd in mixed sorbent systems, the accuracy of the predictions depends directly on the accuracy of the measurements of stability constants for both the aqueous and surface metal-ligand complexes that occur in a system of interest.

Economic Analysis of Immunization Strategies for PRRS Control [corrected].

Porcine reproductive and respiratory syndrome virus (PRRSv) is a swine-specific pathogen that causes significant increases in production costs. When a breeding herd becomes infected, in an attempt to hasten control and elimination of PRRSv, some veterinarians have adopted a strategy called load-close-expose which consists of interrupting replacement pig introductions into the herd for several weeks (herd closure) and exposing the whole herd to a replicating PRRSv to boost herd immunity. Either modified-live virus (MLV) vaccine or live field-virus inoculation (FVI) is used. This study consisted of partial budget analyses to compare MLV to FVI as the exposure method of load-close-expose program to control and eliminate PRRSv from infected breeding herds, and secondly to estimate benefit / cost of vaccinating sow herds preventatively. Under the assumptions used in this study, MLV held economic advantage over FVI. However, sensitivity analysis revealed that decreasing margin over variable costs below $ 47.32, or increasing PRRSv-attributed cost above $18.89 or achieving time-to-stability before 25 weeks resulted in advantage of FVI over MLV. Preventive vaccination of sow herds was beneficial when the frequency of PRRSv infection was at least every 1 year and 9 months [corrected]. The economics of preventative vaccination was minimally affected by cost attributed to field-type PRRSv infection on growing pigs or by the breeding herd productivity level. The models developed and described in this paper provide valuable tools to assist veterinarians in their efforts to control PRRSv.

Regulation of Serum Response Factor and Adiponectin by PPARγ Agonist Docosahexaenoic Acid.

Recent studies indicate that significant health benefits involving the regulation of signaling proteins result from the consumption of omega-3 polyunsaturated fatty acids (ω-3 PUFAs). Serum response factor (SRF) is involved in transcriptional regulation of muscle growth and differentiation. SRF levels are increased in the aging heart muscle. It has not been examined whether SRF is made by adipocytes and whether SRF secretion by adipocytes is modulated by PPARγ agonist DHA. Adiponectin is made exclusively by adipocytes. We and others have previously reported that PUFAs such as DHA increase adiponectin levels and secretion in adipocytes. Here we show that DHA downregulates SRF with a simultaneous upregulation of adiponectin and that both these responses are reversible by PPARγ antagonist. Furthermore, there appears to be a direct relationship between DHA exposure and increased levels of membrane-associated high-density adiponectin, as well as lower levels of membrane-associated SRF. Thus, we find that the levels of SRF and adiponectin are inversely related in response to treatment with PPARγ agonist DHA. Decreased levels of SRF along with increase in membrane-associated adiponectin could in part mediate the health benefits of DHA.

Histoplasma capsulatum and Caenorhabditis elegans: a simple nematode model for an innate immune response to fungal infection.

Histoplasma capsulatum is a primary fungal pathogen of mammals responsible for histoplasmosis. During pathogenesis H. capsulatum yeast proliferate in phagosomes of macrophages. This extensive host/pathogen interaction involves a complex cascade of responses in both organisms. In the mammalian host, infection results in complex branched immunity that is initiated with an innate response and later induces an adaptive response but each response is difficult to resolve during fungal infection. Therefore, in an effort to identify less complex systems and to gain understanding of the host innate response to H. capsulatum, we constructed a mini-host survival assay. With this assay, we found ingestion of virulent Histoplasma capsulatum NAm 1 strain yeasts to be lethal to a Bristol-N2 Caenorhabditis elegans host. The virulent H. capsulatum NAm1 strain shows differential lethality under live/heat-killed infective conditions. Specifically, after ingestion of live yeast lethality is > or = 90% within 48 to 72 h, whereas worms ingesting heat-killed yeast reach equivalent mortality only after 10-14 days. On the other hand, ingestion of live H. capsulatum yeast of the nonvirulent NAm 1 (ura(-)) strain is no more lethal to the nematode than heat-killed yeast. Therefore, C. elegans provides an attractive model for further investigations of the ancient innate immune response during early host/pathogen (H. capsulatum/worm) interaction and pathogenesis.

Impact of a magnetic ion exchange resin on ozone demand and bromate formation during drinking water treatment.

The objective of this research was to examine the impact of a magnetic ion exchange resin (MIEX) on ozone demand and bromate formation in two different ozonated waters at bench scale. The first raw water had a high bromide ion concentration, a high ozone demand, and was highly colored. Based on experimental findings from the first water, the second water was selected as a model water in which more controlled experiments were performed. The waters were treated with the MIEX resin using jar test procedures to find the optimal MIEX dosage based upon the removal of ultraviolet (UV)-absorbing substances, dissolved organic carbon (DOC), and bromide. The optimal resin dosage was chosen for bulk MIEX treatment and subsequent ozonation in a semi-batch reactor. The ozone demand and formation of bromate were analyzed as a function of ozone dosage and dissolved ozone concentration for the MIEX pre-treated water, and compared to the results obtained by ozonating the water without MIEX pre-treatment. The results indicate that pre-treatment of the water with the MIEX resin significantly reduces total organic carbon, DOC, UV absorbance, color, and to some extent, bromide. MIEX pre-treatment of the water prior to ozonation substantially lowered the ozone demand and formation of bromate during subsequent ozonation.

Characterization of an alternative oxidase activity of Histoplasma capsulatum.

Histoplasma capsulatum possesses a branched mitochondrial electron transport chain, with both cyanide-sensitive and -insensitive oxygen-consuming activities. The latter, carried out by a single subunit enzyme termed 'alternative oxidase', is the focus of this report. AOX1 cDNA clones were isolated and direct evidence that the cDNA ORF encodes functional alternative oxidase is reported. Also reported are the generation of an antiserum to the AOX1 protein product, and specific detection in vivo of the mRNA and protein products of the AOX1 gene. Finally, initial studies of regulation of H. capsulatum AOX1 gene expression demonstrated that RNA abundance was increased after hydrogen peroxide-mediated oxidative stress and after inhibition of mitochondrial electron transport enzymes with antimycin A or sodium cyanide. This pattern of regulation is consistent with the hypothesis that alternative oxidase contributes to survival of H. capsulatum after oxidative or metabolic stress and may be important for virulence of this pathogenic organism. The GenBank Accession Nos for the cDNA sequences reported in this paper are AF133236, AF133237 (AOX1).

Redundancy, phylogeny and differential expression of Histoplasma capsulatum catalases.

Histoplasma capsulatum produces an extracellular catalase termed M antigen, which is similar to catalase B of Aspergillus and Emericella species. Evidence is presented here for two additional catalase isozymes in H. capsulatum. Catalase A is highly similar to a large-subunit catalase in Aspergillus and Emericella species, while catalase P is a small-subunit catalase protein with greatest similarity to known peroxisomal catalases of animals and Saccharomycotina yeasts. Complete cDNAs for the CATA and CATP genes (encoding catalases A and P, respectively) were isolated. The transcriptional expression of the H. capsulatum CATA, CATB (M antigen) and CATP genes was assessed by Northern blot hybridizations on total RNA. Results at the transcript levels for these genes are shown for three conditions: cell morphology (mycelial versus yeast phase cells), oxidative stress (in response to a challenge with H(2)O(2)) and carbon source (glucose vs glycerol). Collectively, these results demonstrated regulation of CATA by both cell morphology and oxidative stress, but not by carbon source, and regulation of CATB and CATP by carbon source but not cell morphology or oxidative stress. A phylogenetic analysis of presently available catalase sequences and intron residences was done. The results support a model for evolution of eukaryotic monofunctional catalase genes from prokaryotic genes.