Ultrastructure, Morphology, and Sporogenesis of Pasteuria penetrans

ABSTRACT Pasteuria penetrans is a bacterial parasite of root-knot nematodes that shows great potential as a biocontrol agent. Scanning and transmission electron microscopy were used to study the ultrastructure, morphology, and sporogenesis of four isolates of P. penetrans. The effects of different Meloidogyne spp. and tobacco cultivars on sporangium size and morphology of endospores attached to the cuticle of second-stage juveniles (J2) of root-knot nematodes also were investigated. The P. penetrans isolates and their origins were P-20 from M. arenaria race 1 in Levy County, FL; P-100 from Meloidogyne sp. in Pasco County, FL; B-4 from Pratylenchus scribneri in Seminole County, FL; and P-120 from Meloidogyne spp. in Alachua County, FL. Sporangia of the four isolates were identical morphologically and similar in their dimensions, ranging from 2.39 to 3.42 mum in diameter and from 1.38 to 2.38 mum in height. Different Meloidogyne spp. and tobacco cultivars had no effect on sporangium size. Endospores attached to J2 were visualized in three forms: endospores retaining the sporangium wall, endospores covered with a thin exosporium, and endospores without covering. Sporogenesis of P. penetrans was similar to that of other gram-positive bacteria and generally matched the seven-stage scheme reported for Bacillus thuringiensis.     

Elevating glucose and insulin secretion by carbohydrate formulation diets in late lactation to improve post‐weaning fertility in primiparous sows

Primiparous (P1) sows commonly lose excessive body reserves to meet energy requirements for maintenance and milk production during lactation, and consequently, post‐weaning reproductive performance may be compromised. The present studies determined whether ad libitum feeding a glucogenic carbohydrate diet (CHO) during late lactation could stimulate insulin and glucose secretion (experiment 1) and improve subsequent litter size (experiment 2). For experiment 1, 15 P1 sows, and for experiment 2, 99 P1 sows (198.5 ± 2.7 kg) were allocated randomly according to suckled litter size (≥10 piglets), either to a CHO diet (14.3 MJ DE/kg, 19.8% crude protein) or a standard lactation diet (control; 14.2 DE MJ/kg, 19.5% crude protein) at 8 days before weaning. The CHO diet aimed to provide glucogenic content (extruded wheat, dextrose and sugar) as energy sources instead of fat sources without changing total dietary energy. Pre‐prandial plasma glucose and insulin concentrations were not influenced by treatments. However, post‐prandial plasma glucose and insulin concentrations and their peaks were both higher (p < .05) compared to the control treatment. Body weight loss during lactation was relatively low at 3%–4% for both treatments and did not differ between control and CHO treatments (?7.6 ± 1.6 vs ?5.4 ± 1.2 kg; p > .05). Second litter size was not influenced by diet (p > .05), but the weaning‐to‐mating interval was shorter in CHO sows (p < .05). This study demonstrates that providing an enriched CHO diet in late lactation did influence post‐weaning follicle growth but did not improve subsequent litter size. This may be due to the primiparous sows in this study not experiencing severe negative energy balance and there was no second litter syndrome in this farm which limited the ability of diet to improve sow fertility.     

Applying fire connectivity and centrality measures to mitigate the cheatgrass-fire cycle in the arid West,USA

Context

Strategic placement of fuel treatments across large landscapes is an important step to mitigate the collective effects of fires interacting over broad spatial and temporal extents. On landscapes where highly invasive cheatgrass (Bromus tectorum) is increasing fire activity, such an approach could help maintain landscape resilience.

Objectives

Our objectives are to 1) model and map fire connectivity on a cheatgrass-invaded landscape, as well as the centrality of large cheatgrass patches, in order to inform a landscape fuel treatment (i.e., a network of greenstrips); and 2) evaluate the modeled greenstrip network based on changes to cheatgrass patch centrality.

Methods

Our analysis covers 485-km² on the Kaibab National Forest in Northern Arizona. We apply a circuit-theoretic model of fire connectivity between all pairs of large cheatgrass patches. Based on these results, we calculate a measure of centrality for each patch to inform fuel treatment placement. We evaluate the modeled greenstrip network by comparing the pre- and post-treatment centrality of each patch.

Results

After modeling fire connectivity across the landscape, we identify 25 of 68 large cheatgrass patches with relatively high centrality. When we simulate greenstrips around these focal patches, model results suggest that they are effective in reducing the centrality for at least 19 of the 25 patches.

Conclusions

Fire connectivity models provide robust network centrality measures, which can help generate multiple, landscape fuel treatment alternatives and facilitate on-the-ground decisions. The extension of these methods is well suited for landscape fuels management in other vegetation communities and ecosystems.

     

A simplified method for estimating soil compaction

We describe a simplified model that allows users to explore some of the main aspects of soil compaction. It is intended for use by non-experts, such as students, and is written as an easy-to-use spreadsheet. It estimates soil bulk density under the centre-line of a wheel track from readily available tyre details. The model uses an analytical method to estimate the propagation of stress in the soil. It contains compactibility data for contrasting soils and it accounts for both rebound and recompression realistically. We present examples that show the potential of the model in selecting tyres and wheel systems to minimise compaction.     

Topical anaesthesia alleviates short-term pain of castration and tail docking in lambs

Objective To investigate the effect of a topical anaesthetic formulation on pain alleviation, wound healing and systemic levels of local anaesthetic actives in lambs undergoing castration and tail docking. Design Three placebo-controlled and/or randomised experiments were conducted using three groups of Merino lambs (n = 62, 68 and 19) undergoing routine castration and tail docking. Procedure Surgical castration, with either surgical or hot-iron tail docking, was performed with and without the application of topical anaesthetic (Tri-Solfen®) or placebo. The effects of this procedure were compared with those of rubber ring castration and tail docking, and of the handled but unmarked controls. Wound pain was assessed using calibrated Von-Frey monofilaments over a 4-h period, pain-related behaviour was assessed over 5 h, wound healing was assessed at 14 and 28 days, and the plasma levels of lignocaine and bupivacaine were determined. Results Rapid and up to 4 h primary hyperalgesia developed following surgical castration and tail docking in the untreated and placebo-treated lambs. It was absent in the castration wounds, and significantly reduced in the tail-docking wounds, of the treated lambs. Hot-iron docking was associated with mild and transient secondary hyperalgesia, which was abolished by the topical anaesthesia. There was a significant reduction in pain-related behaviours in treated lambs, which were not significantly different in their behaviour to the sham-operation handled controls. Plasma lignocaine and bupivacaine levels were below the toxic thresholds in all tested lambs. Conclusion Topical anaesthesia alleviates wound pain and significantly reduces pain-related behaviours in lambs undergoing surgical castration plus surgical or hot-iron tail docking, without a negative effect on wound healing or a risk of systemic toxicity.     

The REFLECT Statement: Methods and Processes of Creating Reporting Guidelines for Randomized Controlled Trials for Livestock and Food Safety

The conduct of randomized controlled trials in livestock with production, health, and food-safety outcomes presents unique challenges that might not be adequately reported in trial reports. The objective of this project was to modify the CONSORT (Consolidated Standards of Reporting Trials) statement to reflect the unique aspects of reporting these livestock trials. A 2-day consensus meeting was held on November 18–19, 2008 in Chicago, IL, to achieve the objective. Before the meeting, a Web-based survey was conducted to identify issues for discussion. The 24 attendees were biostatisticians, epidemiologists, food-safety researchers, livestock production specialists, journal editors, assistant editors, and associate editors. Before the meeting, the attendees completed a Web-based survey indicating which CONSORT statement items would need to be modified to address unique issues for livestock trials. The consensus meeting resulted in the production of the REFLECT (Reporting Guidelines for Randomized Control Trials) statement for livestock and food safety and 22-item checklist. Fourteen items were modified from the CONSORT checklist, and an additional subitem was proposed to address challenge trials. The REFLECT statement proposes new terminology, more consistent with common usage in livestock production, to describe study subjects. Evidence was not always available to support modification to or inclusion of an item. The use of the REFLECT statement, which addresses issues unique to livestock trials, should improve the quality of reporting and design for trials reporting production, health, and food-safety outcomes.     

Effect of Antibiotic-containing Extenders on Taylorella equigenitalis Contaminated Semen

Taylorella equigenitalis is a gram-negative coccobacillus and the causative agent of a transmissible venereal disease in horses known as contagious equine metritis. Outbreaks of contagious equine metritis have been documented in various countries since 1977, with the most recent discovery in the United States in December 2008. During disease occurrences, culturing semen samples for T equigenitalis before breeding may help to prevent transmission of this disease; however, little is known about the antimicrobial activity of equine semen extenders against the organism. The purpose of this study was to investigate the infectivity levels of T equigenitalis in three equine semen extenders inoculated with known concentrations of the organism. The semen extenders used for this study included INRA 96, E-Z Mixin BF, and VMDZ. In addition, Timentin was added to INRA 96 at three different concentrations (0.5, 1.0, and 1.5 mg/mL) to investigate possible synergistic effects of antibiotic supplementation of extenders. Results were based on the visual counting of the colonies on chocolate Eugon agar plates. Both INRA 96 (with added Timentin) and VMDZ (as supplied by the manufacturer) significantly reduced the numbers of T equigenitalis isolated from semen extenders as compared with INRA 96 (as supplied by the manufacturer) or the antibiotic free E-Z Mixin BF. Our findings indicate that INRA 96 (with added Timentin) or VMDZ may significantly decrease the growth of T equigenitalis in extended semen; however, it is also important to consider the possible effects of antibiotic supplemented extenders on sperm longevity and fertility in addition to eliminating specific pathogens in semen.     

Assessment of NK cells response to hepatocyte derived chemotactic agents

This project was aimed to examine the NK92 cells response as the CXC chemokine responder cells in rat model of liver disorder and injuries. Hepatocytes were isolated from Sprague-dawly rats and cultured on collagen type 1. Migration of NK92 cells was assessed using a 48 well micro-chemotaxis technique. Transwell chambers were positioned faced up, blocked Medium supernatant (500 microL) obtained from hepatocytes cultures were placed into the lower compartment of each Transwell. The upper compartment was filled with either 500 microL of NK92 cells. After washing, Membrane-attached cells were fixed; stained and Membrane-attached cells were counted by light microscopy and/or by size gating (9-14 microm) with an automated counter system. Human NK92 cells were attracted to recombinant human IP-10 in a concentration and time-dependent manner. NK92 cells also exhibited a chemotactic response to medium harvested from primary hepatocyte cultures. Isolated and cultured hepatocytes express several different chemokines. Although we identified that medium from hepatocyte cultures contains specific chemokines by immunoblotting, there is potential that migration assays detected yet other chemokines and other factors such as complement components. In this report, we demonstrated that hepatocytes expressed factors that were chemoattractive for human NK92 cells and that the factors must interact with the repertoire of receptors responsible for recruitment of these cells.     

The Effects of Growth Factor on Testicular Germ Cell Apoptosis in the Stallion

Apoptosis is necessary for both initiation and control of spermatogenesis; however, an increase in apoptosis can lead to subfertility/infertility in stallions, causing substantial financial loss in the equine industry. The ability of stem cell factor (SCF), leukemia-inhibiting factor (LIF), granulocyte–macrophage colony-stimulating factor (GM-CSF), and estradiol (E₂), alone or in combination, to prevent apoptosis of germ cells in short-term equine testicular cultures was examined. Testicular tissue was sectioned into approximately 2-mm cubes and placed in media-filled culture chambers. Concentrations of SCF (100 ng/mL), LIF (10 ng/mL), GM-CSF (5 ng/mL), and E₂ (10⁻⁹ mol/l) were added alone or in combination to each well. After 6 hours in culture, the tissue was fixed and immunohistochemically (terminal deoxynucleotidyl transferase-mediated nick-end labeling; TUNEL) stained for apoptosis detection. Apoptotic cells per 100 Sertoli cell nuclei within seminiferous tubules were counted until the 500^th Sertoli cell nuclei was reached. This counting procedure was used for each slide. An analysis of variance (ANOVA) with a Tukey's test was used to compare apoptotic rates. In comparison with the control, GM-CSF alone lowered apoptosis by 34.77%. GM-CSF–treated tissue combined with SCF and LIF as well as GM-CSF combined with SCF, LIF, and E₂ reduced apoptosis when compared with the control (37.45% and 44.40%, respectively) or other treatment combinations. GM-CSF alone reduced apoptosis; results suggest possible synergy for the combinations of SCF and LIF with GM-CSF and for E₂ with SCF, LIF, and GM-CSF.