System for quasi‐continuous simultaneous measurement of oxygen diffusion rate and redox potential in soil

Oxygen diffusion rate (ODR) and redox potential (E_H) are quantitative indices representing oxygen availability and redox status in soils, which is valuable information for better understanding causes and effects of soil aeration. Because these indices are spatially and temporally highly variable, continuous measurements and adequate numbers of repetitions are essential for accurate in situ monitoring. Here, we present a new, fully automated recording system for in situ measurements where ODR and E_H are measured at the same platinum electrode. The conflict between electrode polarization for ODR and the resulting biased E_H readings is solved by reducing the polarization time and introducing a recovery interval between two consecutive measurement cycles. The shorter polarization time ensures accurate E_H readings. It also results in moderately overestimated ODR readings, but this can be corrected before data analysis. The recovery interval restricts temporal resolution of the E_H‐ODR data pairs to 8 h. We illustrate the use of the system with measurements in a field experiment in Zürich, Switzerland. ODR curves at different depths ran roughly parallel to the corresponding curves of O₂ concentration in soil air but ODR was much more sensitive to precipitation. Low ODR was a necessary but not a sufficient condition for declining E_H. E_H ran parallel to O₂ concentration in soil air rather than to ODR. The fully automated system allows for time series of replicate measurements in multifactorial field studies with reasonable labor requirements. It may be particularly suitable for studies examining the effects of soil tillage, compaction, and irrigation, where structure‐related soil properties such as porosity, gas permeability, and soil aeration play a dominant role.     

γ-Amino Butyric Acid Control of Arginine Vasopressin Release from the Ewe Hypothalamus In Vitro: Sensitivity to Oestradiol

The present study aims to ascertain the influence of gamma-amino butyric acid (GABA)(A or B) receptors on arginine vasopressin (AVP) release in vitro and determine whether E(2) modulates GABA-AVP interaction. Within 10 min of ewe killing, saggital midline hypothalamic slices (from the anterior preoptic area to the mediobasal hypothalamus along with the median eminence, 2-mm thick, two per ewe) were dissected, placed in oxygenated minimum essential media (MEM)-alpha at 4 degrees C and within 2 h were singly perifused at 37 degrees C with oxygenated MEM-alpha (pH 7.4; flow rate 0.15 ml/min), either with or without E(2) (24 pg/ml). After 4-h equilibration, 10-min fractions were collected for 4 h interposed with a 10-min exposure at 60 min to a specific GABA(A or B) receptor agonist or antagonist at various doses (0.1-10 mm). GABA(A) (muscimol; no E(2), n = 7 perifusion chambers, with E(2), n = 11) or GABA(B) (baclofen; no E(2), n = 8, with E(2), n = 15) agonists (10 mm) did not influence AVP concentrations. However, AVP release increased (p < 0.05) 20-30 min after exposure to 10 mm GABA(A or B) antagonists (bicuculline, no E(2), n = 7: from 4.6 +/- 0.7 to 33.0 +/- 0.4, with E(2), n = 17: from 11.9 +/- 1.4 to 32.8 +/- 6.0; CGP52432, with E(2), n = 14: from 14.0 +/- 2.6 to 28.8 +/- 3.9 pg/ml). At the end of the collection period, hypothalamic slices responded to KCl (100 mm) with AVP efflux (p < 0.05). GABA(B) but not GABA(A) antagonist-stimulated AVP release was enhanced in the presence of E(2). In summary, AVP release is under the inhibitory influence of GABA input with further potentiation by E(2) through GABA(B) receptors in vitro.     

Immunolocalization of Aquaporins 1 and 9 in the Ram Efferent Ducts and Epididymis

Aquaporins (AQPs) are essential membrane protein channels for the transport of water across membranes. Fluid movement in the epididymis is important for modulation of the luminal environment, in which sperm mature and reside. This study was designed to understand the morphology and localization of AQPs in ram efferent ducts (ED) and epididymis. For this purpose, the epididymis of seven animals were removed for histologic and immunohistochemical analyses. AQP1 immunoreactivity was observed in the apex of the ED, and AQP9 was found adjacent to the nuclei of the epithelial cells of the ED. The epithelial lining of ram epididymis is pseudostratified columnar and presents principal, basal, apical and narrow cells. In the initial segment (IS), a moderate reaction for AQP1 was observed in the apical cytoplasm of epithelial cells. An intense reactivity for AQP1 was noted over the microvilli of principal cells and in spermatozoa in the caput. In the corpus and cauda, AQP1 was noted only over the endothelial cells of vascular channels located in intertubular spaces. A weak‐to‐moderate reaction for AQP9 was observed in the nuclei of epithelial cells in the IS, caput and corpus of the epididymis. In the cauda, an intense reaction to AQP9 was observed in the epithelial border. In the IS, caput and corpus, the reactivity for AQP9 differed from those observed in domestic animals. The cauda showed a pattern similar to that previously described. These results indicate that AQPs 1 and 9 have reversed locations and roles in rams, suggesting activity variations related with fluid and solute absorption throughout the epididymis.     

Kisspeptin,c‐Fos and CRFR Type 2 Co‐expression in the Hypothalamus after Insulin‐Induced Hypoglycaemia

Normal reproductive function is dependent upon availability of glucose and insulin‐induced hypoglycaemia is a metabolic stressor known to disrupt the ovine oestrous cycle. We have recently shown that IIH has the ability to delay the LH surge of intact ewes. In the present study, we examined brain tissue to determine: (i) which hypothalamic regions are activated with respect to IIH and (ii) the effect of IIH on kisspeptin cell activation and CRFR type 2 immunoreactivity, all of which may be involved in disruptive mechanisms. Follicular phases were synchronized with progesterone vaginal pessaries and at 28 h after progesterone withdrawal (PW), animals received saline (n = 6) or insulin (4 IU/kg; n = 5) and were subsequently killed at 31 h after PW (i.e., 3 h after insulin administration). Peripheral hormone concentrations were evaluated, and hypothalamic sections were immunostained for either kisspeptin and c‐Fos (a marker of neuronal activation) or CRFR type 2. Within 3 h of treatment, cortisol concentrations had increased whereas plasma oestradiol concentrations decreased in peripheral plasma (p < 0.05 for both). In the arcuate nucleus (ARC), insulin‐treated ewes had an increased expression of c‐Fos. Furthermore, the percentage of kisspeptin cells co‐expressing c‐Fos increased in the ARC (from 11 to 51%; p < 0.05), but there was no change in the medial pre‐optic area (mPOA; 14 vs 19%). CRFR type 2 expression in the lower part of the ARC and the median eminence was not altered by insulin treatment. Thus, disruption of the LH surge after IIH in the follicular phase is not associated with decreased kisspeptin cell activation or an increase in CRFR type 2 in the ARC but may involve other cell types located in the ARC nucleus which are activated in response to IIH.     

Nutritional importance of pyrroloquinoline quinone

Mice fed a chemically defined diet devoid of pyrroloquinoline quinone (PQQ) grew poorly, failed to reproduce, and became osteolathyritic. Moreover, severely affected mice had friable skin, skin collagen that was readily extractable into neutral salt solutions, and decreased lysyl oxidase. The identification of functional defects in connective tissue and the growth retardation associated with PQQ deprivation suggest that PQQ plays a fundamental role as a growth factor or vitamin.     

Assessment of producer conducted antemortem inspection of market pigs in Australia

OBJECTIVE: A field trial was conducted to evaluate the effectiveness of on-farm antemortem inspection performed by Australian pig producers in terms of proficiency in detecting unhealthy animals (suspects), the impact on food safety and animal welfare outcomes. ANIMALS: A total of 64 trial groups of market pigs (n = 10,703) were dispatched from 20 herds to three abattoirs in three states. PROCEDURE: All producers and abattoir antemortem inspectors were trained in antemortem inspection prior to the trial, using a standardised set of antemortem criteria. Study 1 consisted of three comparisons: on-farm versus abattoir antemortem inspections on the same pigs, transport effects in suspect and normal pigs, and meat rejection and carcase disposition in suspect and normal pigs. Study 2 compared Salmonella spp. status of suspect and normal pigs. Studies 3 and 4 were conducted to assist the interpretation of the results of on-farm versus abattoir inspection. Antemortem inspection results of three producers and one abattoir antemortem inspector were compared with those of a reference inspector (Gold Standard). RESULTS: Of the pigs examined, 2.12% (95% CI: [1.84%, 2.42%]) were classified as suspect at on-farm antemortem inspection compared to only 0.14% at abattoir antemortem inspection. Forty one percent of pigs classified as suspect on-farm had locomotor problems (arthritis, lameness and foot abscess). Compared to normal pigs, suspect pigs had a relative risk of 67.7 (P < 0.0001) of suffering transport injuries. Meat rejection amounted to the equivalent of 9.68% of suspect carcases, compared to the equivalent of 0.35% of normal carcases. The isolation of Salmonella species from caecal content from 20.9% of 67 suspect pigs and 18.8% of 133 control pigs (two matched healthy pigs from the same batch for each suspect pig) were not significantly different. When compared with the reference inspector on-farm, three producers classified abnormalities in 2495 pigs with high specificity (0.98), acceptable sensitivity (0.82) and kappa (0.57). CONCLUSIONS: With training, producer sensitivity in antemortem inspection may be high and more proficient than abattoir antemortem inspection. Use of producer inspections may have benefits for animal welfare and chain efficiency, but not food safety.