Evaluation of an enzyme-linked immunosorbent assay for the detection of Mycoplasma hyopneumoniae antibody in porcine serum

An enzyme-linked immunosorbent assay (ELISA) for detecting antibody to Mycoplasma hyopneumoniae in porcine serum is described. The results are presented as an ELISA ratio, calculated by dividing the absorbance of the test sample by the mean absorbance of control negative sera. In known infected pigs, the ELISA ratio was highest when the serum concentration applied to the ELISA plate was diluted 1 in 20 in PBS - Tween. Mean ELISA ratios ranged from 1.2 +/- 0.3 for pigs without porcine enzootic pneumonia (PEP) lesions to 5.5 +/- 1.5 for pigs observed with a PEP lesion reacting positively with immunofluorescent histopathology. Pigs observed with typical PEP lesions at slaughter, but not confirmed by immunofluorescent histopathology had a mean ELISA ratio of 4.9 +/- 1.7. The ELISA was highly sensitive (95.6%) and specific (98.8%) when pig sera from commercial piggeries of known M hyopneumoniae infection status were assessed. No cross-reactivity with serum from a pig hyperimmunised with killed M flocculare was detected, and reactivity with serum from another pig hyperimmunised with killed M hyorhinis showed only weak cross-reactivity, which failed to reach the ELISA positive threshold (ELISA ratio 3) for M hyopneumoniae.     

Estimating evapotranspiration in semi-arid rangelands: connecting reference to actual evapotranspiration and the role of soil evaporation

In a context of water scarcity, efforts to increase landscape production should focus on improving water productivity. This requires an appreciation of the various components of evapotranspiration (ET), including soil evaporation (E_s) because the latter reflects ‘unproductive’ water loss. Both complex and simple algorithms have been developed to determine ET. In data scarce areas, developing and testing parsimonious algorithms is useful. This study sought to improve a simple single layer ET model by incorporating an E_s component. Empirical methods were also explored to predict ET from vegetation indices (VIs), leaf area index (LAI) and reference evapotranspiration (ET₀). A large aperture scintillometer and an eddy covariance (EC) system were used to validate the proposed algorithm at three sites over Grasslands and Albany Thicket biomes in the Eastern Cape, South Africa. There was good agreement between the observed and predicted ET with RMSE of 0.30–0.58 mm d^?1 when average daily observed ET was 0.43–3.24 mm. The VIs had moderate correlations with the observed data due to the significant role played by E_s (65%–84%) across the sites and stomatal conductance at the Albany Thicket site. The simple algorithms developed would make determining ET easier in data scarce regions.     

In vitro Developmental Competence of Adult Sheep Oocytes Treated with Roscovitine

The efficiency of in vitro sheep embryo production is still low compared to that observed in vivo and in other species. In this context, meiotic inhibition strategies emerged as a promising alternative to improve this biotechnology. So, this study aimed to evaluate, for the first time, the effects of roscovitine on in vitro maturation of sheep oocytes and their subsequent embryo development. For this, cumulus–oocyte complexes (COCs) were cultured for 6 h in the presence (Rosco) or absence (Control) of 75 μm roscovitine and, subsequently, in vitro matured (IVM) for 18 h with gonadotropins. At 0 (Immature), 6 and 24 h of culture, the nuclear status of oocytes was evaluated by Hoechst staining. Embryo cleavage and blastocyst formation were recorded 30 h after in vitro fertilization and on day 7 of culture, respectively. Blastocyst quality was evaluated by differential staining. At 6 h, the GV rate in the Rosco treatment (93.8%) was similar to that observed in the Immature oocytes (94.9%) and significantly higher compared to Control (41.3%). After IVM for 18 h, a high and similar proportion of oocytes from Rosco (93.6%) and Control (88.4%) reached the MII stage. In both treatments, approximately 70% of oocytes cleaved and 50% of them developed up to blastocyst. The mean percentage of blastocyst cells, embryoblast, trophoblast and pyknosis did also not differ between Control and Rosco. In conclusion, roscovitine, at the studied experimental conditions, was efficient to reversibly inhibit the meiosis of adult sheep oocytes without detrimental effect on development and quality of the in vitro produced embryos.     

Effects of cinnamaldehyde, garlic and juniper berry essential oils on rumen fermentation, blood metabolites, growth performance, and carcass characteristics of growing lambs

The objective of this study was to examine the effects of cinnamaldehyde (CDH), garlic (GAR) and juniper berry (JUN) essential oils (200 mg/kg of DM) on performance and carcass characteristics of lambs fed a barley-based concentrate diet ad libitum. For this purpose, 40 ewes' lambs (23.5 ± 1.11 kg initial live weight, LW) were used in a random block design over a 13-week period. Feeding CDH, GAR or JUN did not affect dry matter intake (DMI) but the average daily gain (ADG) of lambs supplemented with CDH and JUN was higher (P = 0.002) as compared to lambs fed GAR or the control diet. Feed conversion (DMI/ADG) was numerically improved when lambs were fed CDH (4.8) and JUN (4.7) compared to those fed GAR (5.2) or the control diet (5.3). There were no effects of feed additives on ruminal pH and concentrations of ammonia and total VFA. Serum concentrations of glycerol and total glycerides were lower and higher (P ≤ 0.03) in lambs fed CDH or JUN respectively, as compared to lambs fed GAR or the control diet. Hot dressed carcass weight was similar among treatments (23.7 ± 0.75 kg; P = 0.18) whereas saleable meat tended (P = 0.13) to increase (+ 9%) in lambs fed CDH and JUN compared to those fed GAR or the control diet. Feeding CDH, GAR or JUN had little effect on the overall fatty acid composition of back fat and liver and only minor effects on meat flavour characteristics.     

Germination of Eucalyptus sieberi L. Johnson seeds. II. Internal water relations

At 15-20 degrees C, seeds of Eucalyptus sieberi L. Johnson need to maintain a water content equal to 30% or more of dry weight for approximately 60 h in order to germinate. Several shorter periods of imbibition at 30% water content separated by between 6 and 120 h desiccation were as effective as an unintenupted imbibition in bringing seeds to the point of germination. Even after 8 months' desiccation an earlier imbibition shortened the time required for the completion of germination, although it did not hasten the onset of germination. The water potential of seeds sufficiently hydrated to germinate was about -0.6 MPa, whereas that of the embryo was only about -4.5 MPa. The difference was caused by the inhibition of water uptake by the inner integument of the seed coat. During germination, reserve protein in a small group of cells above the collet was hydrolyzed. Subsequent expansion of these cells caused rupture of the inner integument thereby allowing further water uptake by the embryo. The balance of water absorption and evaporative loss by the outer integument was such that most seeds on a moist substrate but exposed to dry air (6.5-8 x 10(-4) MPa) did not maintain the 30% water content necessary for germination. Thus although able to germinate on seedbeds subject to intermittent drying, E. sieberi seeds have the capacity to avoid germination except under humid conditions.     

Modeling production of antifungal compounds and their role in biocontrol product inhibitory activity

Partial least squares (PLS) regression modeling was used to relate the antifungal activity of Bacillus subtilis solid-state fermentation extracts to the individual high-performance liquid chromatography (HPLC) peaks from those extracts. A model was developed that predicted bioassay inhibition based on the extract HPLC profile (R(2) = 0.99). Concentrations of the members of the antifungal lipopeptide families iturin A and fengycin were found to correlate positively with extract inhibition, but a peak with unidentified chemical composition (designated as peak 48) showed the strongest correlation with extract inhibition. HPLC data were used to construct models for the production of iturin A, fengycin, and peak 48 as a function of the substrate moisture content, incubator temperature, and aeration rate in the solid-state bioreactors. Maximum production of all compounds occurred at the highest moisture content (1.7 g/g dry basis) and lowest incubator temperature (19 degrees C) tested. Optimal aeration rates for the production of the two known lipopeptides and peak 48 were 0.1 and 1.5 L/min, respectively.     

Prior determination of baseline minimum alveolar concentration (MAC) of isoflurane does not influence the effect of ketamine on MAC in rabbits

The objective of this study was to compare the effect on the minimum alveolar concentration (MAC) of isoflurane when ketamine was administered either after or without prior determination of the baseline MAC of isoflurane in rabbits. Using a prospective randomized crossover study, 8 adult, female New Zealand rabbits were allocated to 2 treatment groups. Anesthesia was induced and maintained with isoflurane. Group 1 (same-day determination) had the MAC-sparing effect of ketamine [1 mg/kg bodyweight (BW) bolus followed by a constant rate infusion (CRI) of 40 μg/kg BW per min, given by intravenous (IV)], which was determined after the baseline MAC of isoflurane was determined beforehand. A third MAC determination was started 30 min after stopping the CRI. Group 2 (separate-day determination) had the MAC-sparing effect of ketamine determined without previous determination of the baseline MAC of isoflurane. A second MAC determination was started 30 min after stopping the CRI. In group 1, the MAC of isoflurane (2.15 ± 0.09%) was significantly decreased by ketamine (1.63 ± 0.07%). After stopping the CRI, the MAC was significantly less (2.04 ± 0.11%) than the baseline MAC of isoflurane and significantly greater than the MAC during the CRI. In group 2, ketamine decreased isoflurane MAC (1.53 ± 0.22%) and the MAC increased significantly (1.94 ± 0.25%) after stopping the CRI. Minimum alveolar concentration (MAC) values did not differ significantly between the groups either during ketamine administration or after stopping ketamine. Under the study conditions, prior determination of the baseline isoflurane MAC did not alter the effect of ketamine on MAC. Both methods of determining MAC seemed to be valid for research purposes.