Evaluation and application of a paper-based device for the determination of reactive phosphate in soil solution.

The evaluation and validation of a new low-cost microfluidic paper-based analytical device (µPAD) for the determination of reactive phosphate in soil solution is described. This device allows up to 15 replicate measurements of reactive phosphate on one credit card-sized device and requires only a desktop or hand scanner for signal detection and quantification. The proposed method showed a linear response between 0.1 and 1.0 mg L and between 1.0 and 10.0 mg L P with a limit of detection of 0.05 mg L P. When applied to the analysis of soil solution, there was excellent agreement between results obtained using the µPAD and those obtained by a reference spectrophotometric method, as indicated by the following regression equation: [P] = (0.997 ± 0.005)[P] - (0.020 ± 0.008) ( = 0.997; = 110). It was found that the ambient temperature storage stability of the µPAD could be extended to 15 d by incorporating a removable polymeric interleaving sheet between the adjacent paper layers of the device. The observed sensitivity of the µPADs to sunlight, which was manifested by photoreduction of the chromogenic molybdate reagent used in the assay, was overcome by preparing the µPADs with an ultraviolet-filtering laminating material. The proposed method is rapid, with a reaction time of only 10 min, is easy to perform, and is suitable for application in the field.

Risk of kidney stones with surgical intervention in living kidney donors.

A kidney stone in a person with a solitary kidney requires urgent attention, which may result in surgical and/or hospital attention. We conducted a matched retrospective cohort study to determine if living kidney donors compared to healthy nondonors have a higher risk of: (i) kidney stones with surgical intervention, and (ii) hospital encounters for kidney stones. We reviewed all predonation charts for living kidney donations from 1992 to 2009 at five major transplant centers in Ontario, Canada, and linked this information to healthcare databases. We selected nondonors from the healthiest segment of the general population and matched 10 nondonors to every donor. Of the 2019 donors and 20 190 nondonors, none had evidence of kidney stones prior to cohort entry. Median follow-up time was 8.4 years (maximum 19.7 years; loss to follow-up <7%). There was no difference in the rate of kidney stones with surgical intervention in donors compared to nondonors (8.3 vs. 9.7 events/10 000 person-years; rate ratio 0.85; 95% confidence interval [CI] 0.47-1.53). Similarly there was no difference in the rate of hospital encounters for kidney stones (12.1 vs. 16.1 events/10 000 person-years; rate ratio 0.75; 95% CI 0.45-1.24). These interim results are reassuring for the safety of living kidney donation.

Endometrial explant culture to study the response of equine endometrium to insemination.

Mating-induced endometritis (MIE) is ubiquitous in the horse after natural mating and artificial insemination with frozen/thawed semen causing the most aggressive response. The majority of mares eliminate MIE 24-48 h after insemination. An endometrial explant culture was tested as a potential in vitro exemplar for sperm-induced MIE. Endometrial prostaglandin F(2alpha) (PGF(2alpha)) secretion and expression of interleukin-8 (IL-8) were used as markers of inflammation. Endometrial explants were cultured from uteri collected from follicular phase mares. Explants were challenged with 1 or 10 x 10(6) sperm/ml frozen/thawed semen, chilled semen, washed sperm or seminal plasma. Medium was collected 24 and 72 h after challenge and assayed for PGF(2alpha) by radioimmunoassay. Treatment of endometrial explants with frozen/thawed, chilled semen or washed sperm did not change the secretion of PGF(2alpha) compared with untreated controls. However, 24 h after challenge cultured explants expressed IL-8. The in vitro endometrial explant system did not represent the in vivo response to semen when PGF(2alpha) was used as a marker of inflammation, yet the use of gene expression as an inflammatory marker warrants further investigation.

Polychlorinated biphenyls: metabolic behavior of pure isomers in pigeons, rats, and brook trout.

The metabolic behavior of pure mono-, di-, tetra-, and hexachlorobiphenyl isomers in pigeons, rats, and brook trout was investigated. Excreta from these animals were extracted and examined by chromatographic and mass spectrometric techniques. The results showed conversion of the 4-chloro-, 4,4'-dichloro-, and 2,2',5,5'-tetrachlorobiphenyl isomers into monohydroxylated derivatives by the rat and pigeon whereas no hydroxymetabolites were detected in the excreta of the brook trout. No hydroxylated products of 2,2',4,4',5,5'-hexachlorobiphenyl were detected in the excreta of pigeons, rats, or brook trout.

Direct Exports of Phosphorus from Fertilizers Applied to Grazed Pastures.

Since its discovery in 1669, phosphorus (P) in the form of fertilizer has become an essential input for many agroecosystems. By introducing a concentrated P source, fertilizers increase short-term P export potential soon after their application and longer-term export potential by increasing soil fertility (legacy P). The 4R concept was developed to help mitigate P exports from the fertilizers that sustain agricultural productivity. This review investigates the factors affecting P exports soon after the application of mineral fertilizers to pasture-based grazing systems and studies quantifying its potential impact in different systems, with an emphasis on Australasia. Initially, P fertilizers and reactions that might affect their short-term P export potential are reviewed, along with P transport pathways, the forms of P exported from grazing systems, factors affecting P mobilization into water, and studies demonstrating the possible short-term effects of fertilizer application on P exports. Using that foundation, we review studies quantifying the short-term impact of fertilizer application in different regions; they show that under poor management, recently applied fertilizer can contribute a considerable proportion (30-80%) of total farm P exports in drainage, but when fertilizer is well-managed, that figure is expected to be <10%. We then use three model systems of varying hydrology that are common to Australasia to demonstrate the principles for selecting fertilizers that are likely to minimize P exports soon after their application.

The effects of Arcanobacterium pyogenes on endometrial function in vitro, and on uterine and ovarian function in vivo.

Uterine bacterial infection after parturition causes endometritis, perturbs ovarian function and leads to infertility in cattle. Although endometritis is caused by mixed infections, endometrial pathology is associated with the presence of Arcanobacterium pyogenes. The aims of the present study were to determine the effects of A. pyogenes on endometrial function in vitro, and on uterine and ovarian function in vivo. Heat-killed A. pyogenes did not affect the production of prostaglandin F2alpha (PGF) or prostaglandin E(2) (PGE) from endometrial explants, or purified populations of endometrial epithelial or stromal cells. However, the explants produced more PGF and PGE than controls when treated with a bacteria-free filtrate (BFF) cultured from A. pyogenes. Similarly, BFF stimulated PGF and PGE production by epithelial and stromal cells, respectively. So, BFF or control PBS was infused into the uterus of heifers (n=7 per group) for 8 days, starting the day after estrus. Emergence of the follicle wave, dominant follicle or corpus luteum diameter, and peripheral plasma FSH, LH, estradiol, progesterone, PGFM, or acute phase protein concentrations were unaffected by the BFF infusion. In the live animal it is likely that the intact uterine mucosa limits the exposure of the endometrial cells to the exotoxin of A. pyogenes, whereas the cells are readily exposed to the toxin in vitro.

Sources of phosphorus lost from a grazed pasture receiving simulated rainfall.

Nutrients exported from grazing systems contribute to eutrophication of surface waters. In this study the contributions of soil, pasture-plants, and dung to P exports in overland flow were compared using simulated rainfall. The treatments were (i) grazed pasture-plants (isolated from soil by application of petrolatum to the soil surface), (ii) grazed pasture-plants and supporting soil, (iii) grazed pasture-plants and soil and treading, and (iv) grazed pasture-plants and soil and treading and dung. In general, dissolved reactive P (DRP) accounted for the majority of the P exported and P losses decreased in the order: treading and dung treatment>treading>pasture-plants and soil>pasture-plants. Very little dissolved organic P was lost in overland flow and the effects of treading diminished with time. Over a normal grazing cycle (30 d), the portion of P lost from pasture-plants was approximately half that lost from pasture-plants and soil, one-third that lost from treaded pasture-plants and soil, and one-quarter that lost from treaded pasture-plants, soil, and dung. The DRP in the pasture-plants treatment was approximately half that in the pasture-plants and soil treatment and suggests that a significant portion of the P exported from these systems is derived directly from pasture-plants. Due to higher proportions of particulate P (PP) in the treaded and dung treatments, DRP accounted for less of total P than in the pasture-plants and pasture-plants and soil treatments. Lower infiltration capacities probably caused by mechanical disaggregation at the soil surface are consistent with the higher proportions of PP in the treading treatments. These results were used to estimate P exports from a field trial site in Southland, New Zealand. The results suggested that P export attributable to fertilizer, dung, pasture-plants, and soil components were approximately 10, 30, 20, and 40%, respectively. These results suggest that since 90% of the P exports are derived from the soil-plant system and dung returns, managements to lessen P exports should continue to focus on maintaining soil P within the optimal range for pasture-plant production and maintaining soil surface properties that maximize infiltration and minimize overland flow.

Steroidogenic factor-1 regulates the rate of proliferation of normal and neoplastic rat ovarian surface epithelial cells in vitro.

Steroidogenic factor-1 (SF-1) is a transcription factor that is expressed by many cell types within the ovary and has been shown to inhibit granulosa cell proliferation. The present studies were designed to determine whether: 1) SF-1 is expressed by primary and transformed rat ovarian surface epithelial cells (i.e. ROSE cells); and 2) SF-1 expression effects the proliferation of both normal and neoplastic ROSE cells. These studies used immature, gonadotropin-primed and mature rat ovaries, as well as ROSE-179 cells from early passages (EP) and late passages (LP), T-sv-40 transformed ROSE cells, and T-ras transformed ROSE cells. In situ hybridization studies demonstrated that SF-1 was detected in the surface epithelium of rat ovaries, independent of age or gonadotropin treatment. Further, Northern blot and quantitative in situ hybridization studies revealed that significant amounts of SF-1 messenger RNA (mRNA) were present in EP-ROSE-179 cells but not in the other cell lines. Interestingly, EP-ROSE-179 cells proliferated at a significantly slower rate than the other cell lines. Further, SF-1 mRNA levels were higher in EP-ROSE-179 cells in the G0/G1 stage than in the S-, G2/M stage of the cell cycle. These observations suggest that a cause and effect relationship exists between the level of SF-1 expression and cell proliferation. To test this hypothesis, LP, T-sv-40, and T-ras ROSE cells were transfected with either control vector or SF-1 expression vector. Forty-eight hours after transfection, SF-1 expression was assessed by in situ hybridization, and the fold increase in cell number/24 h was determined. For each cell line, about 30% of the cells were successfully transfected. The fold increase in the number of cells observed after transfection with the SF-1 expression vector was significantly less than the increase in cell number after transfection with the control vector (P < 0.05). To confirm that the forced expression of SF-1 prevented proliferation, LP cells were cotransfected with a green fluorescent protein (GFP) expression vector and either control vector or SF-1 expression vector. This study demonstrated that virtually none of the GFP/SF-1-transfected cells proliferated over a 24-h period, whereas GFP/Control vector-transfected cells proliferated. Further, approximately 40% of the GFP/SF-1-transfected cells underwent apoptosis after 24 h of culture in serum-supplemented medium. These data demonstrate that: 1) normal ovarian surface epithelial cells express SF-1; 2) SF-1 is also expressed by EP-ROSE-179 cells, but its expression seems to be suppressed when the cells enter the cell cycle; 3) LP-, T-sv, and T-ras ROSE cells do not express SF-1 mRNA; and 4) the inability to express SF-1 is associated with an increase in cell proliferation. Finally, forced SF-1 expression interferes with serum-induced proliferation and leads to apoptosis.

A pneumatic device for attaching wing bands to day-old chicks.

A device for attaching aluminum bands to the wings of day-old chicks is described and illustrated. It is powered with compressed air and utilizes both hand and foot operated controls. Using this device, an operator can apply wing bands without an assistant at rates up to 300 chicks/hour.

Some aspects of the composition of avian ascitic fluid.

Ascitic fluid from male Single Comb White Leghorn chickens fed a diet containing 20% by weight rapeseed oil rich (51.6 weight percent) in erucic (22:1n9) acid contained 2.84 +/- .09 g/dl protein and 2.55 +/- .22 micrograms/ml DNA. A characteristic feature of ascitic fluid as compared to liver and plasma was its high polyunsaturates, low saturates, and low saturate to unsaturate ratio. In general, the fatty acid profile of ascitic fluid is more comparable to the fatty acid profile of plasma lipids than to that of liver lipids. These data suggest that there is no direct absorption of fat and transfer to the ascitic fluid.

Growth, heart weight, cardiac lipid, and pathology of chickens fed soybean oil or oil extracted from different rapeseed cultivars.

A total of 384 male Single Comb White Leghorn chicks of the Hyline strain were randomized in 24 Petersime battery units of 16 birds per unit. Four replicate pens were fed either a basal (control) diet containing no added oil or a diet supplemented with 20% by weight of either soybean oil or rapeseed oil (RSO) from cultivars Tower, Candle, Regular, or R-500. Levels of crucic acid (22:1) in the RSO's ranged from .1 to 51.6%. Four birds from each unit were killed at 4, 8, 12, and 16 weeks for cardiac lipid analysis and pathology. Feeding RSO had no consistent effect on body weights, heart weights, or the heart-to-body weight ratio. Only the diet containing R-500 (51.6% 22:1) consistently increased cardiac lipid levels as measured gravimetrically. Maximum deposition of the long-chain monoenes (22:1) and eicosenoic acids (20:1) occurred in the cardiac lipids by 8 weeks, and the levels remained high throughout the 16-week trial. The relative concentration of these long-chain monoenes in the cardiac lipids was positively correlated to the dietary levels of these acids. The incidence of lesions found in heart, liver, and skeletal muscle was significantly higher in chickens fed RSO's high in 22:1 than in chickens fed control or soybean oil, and the incidence significantly correlated to the level of 22:1 in the diet. The incidence of heart and muscle lesions and sinusoidal distention in chickens fed RSO's low in 22:1 was not significantly different from that of chickens fed the control diet or soybean oil, but the incidence of hepatic necrosis was significantly higher for chickens fed RSO's low in 22:1 compared to those fed the control diet or soybean oil.

Pathological changes associated with the feeding of soybean oil or oil extracted from different rapeseed cultivars to single comb white Leghorn cockerels.

A total of 384 Single Comb White Leghorn cockerels of the Hyline strain were fed either a basal (control) diet containing no added oil or a diet supplemented with 20% by weight of soybean oil, rapeseed oil from cultivars Tower, Candle, a mixture of Echo and Arlo high erucic acid rapeseed (HEAR), or R-500. Levels of erucic acid (22:1) in the rapeseed diets varied from .03 to 10.31%. Three birds from each unit were killed at 28, 56, 84, and 112 days and a wide range of tissues were examined histologically. A number of birds in all dietary groups had healed lesions of avian encephalomalacia. Two cockerels fed HEAR oil and 12 fed R-500 developed marked ascites, firm shrunken livers, hydropericardium, and cachectic muscular atrophy. Marked periacinar necrosis was present in birds dying with ascites. A significantly higher number of birds fed the rapeseed oils developed hepatic sinusoidal distention than birds fed the basal or soybean oil diets. Degenerative myocardial changes were seen only in R-500 and HEAR oil-fed birds. This, coupled with hepatic changes, producing shrunken firm livers, led to development of ascites, cachectic muscular atrophy, and periacinar hepatic necrosis. These changes were probably produced by the high erucic acid content of the HEAR and R-500 oils and the resultant grossly imbalanced diet.

Equine endometrial cytology and bacteriology: effectiveness for predicting live foaling rates.

Endometritis is an important cause of sub-fertility in mares. The critical indicator of reproductive success and financial return for commercial studs is live foaling rate. Endometrial bacteriology and/or cytology are used to diagnose endometritis and thus identify mares at risk of early embryonic death. However, mares with endometritis may conceive but then abort in late gestation. The aims of this study were to establish, as part of a standard breeding examination (1) whether a threshold percentage of uterine polymorphonuclear neutrophils (PMNs) exists above which a significant reduction in live foaling rate is evident; (2) the relationship of a positive bacteriology result to live foaling rate, and (3) the relationship of a combination of positive cytology and bacteriology result to live foaling rate. Guarded endometrial swabs (n=2660) were collected from 1621 Thoroughbred mares on 17 commercial stud farms by five veterinarians during a single breeding season. All mares were included regardless of age, history or parity. Cytological and bacteriological analyses were performed on each swab and subsequent live foaling rates recoded. Data were analysed by comparing 0%, ≥ 1%, ≥ 2%, ≥ 5% or ≥ 25% PMNs of total cells counted, or categories of bacterial growth to live foaling rates, using Pearson's chi-squared test. A threshold value of ≥ 1% PMNs, culture of a single bacterial isolate and a combination of both these parameters were associated with significantly reduced live foaling rates. Positive cytology alone, positive bacterial culture alone, or combined positive cytology and bacteriology were equally indicative of the likelihood of a mare producing a live foal.

Markers of the uterine innate immune response of the mare.

Reproductive efficiency in mares is low and persistent mating-induced endometritis (PMIE) is an important cause of subfertility. Mating-induced endometritis (MIE) an obligate precursor to PMIE, is a ubiquitous, transient inflammatory response to the presence of sperm, seminal components and pathogens. However, the specific inflammatory pathways that derive from MIE and that may also be precursors to PMIE are not clear. The ability to identify and measure robust, repeatable markers of inflammation integral to MIE may be key to understanding the progression to PMIE. The aim of the study was to (i) refine a protocol for inducing MIE and in doing so test a range of cellular and molecular parameters as valid markers of MIE to facilitate future studies of mares susceptible to PMIE (ii) concurrently identify those parameters with potential as inflammatory indicators during MIE to inform and enhance early treatment regimens in practice. Mating-induced endometritis was induced in pony mares using a stringent protocol; mares were treated intrauterine with frozen/thawed semen (n = 5; FTS) or frozen/thawed extender (n = 6: FTEx). The parameters tested were measured before treatment were compared to samples collected at strategic time points after treatment: uterine cytology using cytological (at 8, 16, 24, 48 and 72 h after treatment) or histological analysis (at 24 and 72 h); uterine bacteriology (at 24 and 72 h); secretion of prostaglandin F(2alpha) (PGF(2alpha); at 8, 16, 24, 48 and 72 h); peripheral concentrations of serum amyloid A (SAA; at 24h); endometrial mRNA gene expression, focussing upon IL8 and TLR4, as examples of genes pertinent to inflammation (at 24 h). Uterine neutrophil cell numbers in both treatment groups increased at 8 (P < 0.001), 16 (P < 0.01) and 24 (P < 0.01) h after insemination, indicative of MIE and distinguished between different treatments because neutrophil numbers were greater from FTS mares than FTEx mares 8h after challenge. Uterine neutrophil cell numbers, assessed by histology, increased (P < 0.001) 24 and 72 h after treatment. Prostaglandin F(2alpha) concentrations increased (P < 0.05) 16 h after treatments, while SAA concentrations and bacterial growth scores were not significantly different after treatment. Endometrium from pony mares expressed mRNA for IL8 and TLR4 but expression was not altered after insemination. The protocol induced MIE, as confirmed by uterine cytology and maybe used hereafter as a repeatable and robust method for studying immune mechanisms that underlie MIE and so may aid the understanding of progression to persistent inflammation. It can be concluded that of the range of parameters tested, neutrophil cell numbers by cytological analysis and PGF(2alpha) were regarded as the most accurate markers of inflammation during MIE and important for use in practice.

Volume imaging: three-dimensional appreciation of the fetal head and face.

Quasi-three-dimensional volume imaging provides an inexpensive means of evaluating the usefulness of three-dimensional imaging. The technique works most efficiently with water-skin interfaces and therefore we investigated its application in obstetrical ultrasonography. Three-dimensional perspectives of the normal and abnormal fetal head and face were spectacular and at times provided more information than the two-dimensional images. The ability of an inexperienced observer to interpret the three-dimensional image more easily may have a role in training sonographers and counseling parents whose fetuses have structural defects. Volume imaging has certain limitations and can only be used as a complementary technique.