Expression of Genes Associated with Apoptosis in the Porcine Corpus Luteum During the Oestrous Cycle

The corpus luteum (CL) of the pig lacks luteolytic sensitivity (LS) to prostaglandin (PG) F‐2α until after day 12 of the oestrous cycle, but the mechanisms underlying this phenomenon are poorly understood. As luteolysis involves apoptosis, we hypothesized that critical apoptotic proteins may be deficient in CLs that lack LS. The specific aim of these studies was to examine mRNA expression and protein levels of apoptosis genes/proteins (BAX/Bax, BCLX/Bcl‐x, CASP3/Caspase‐3, CASP8/Caspase‐8, NFΚB1/NFκB, TP53/p53) in porcine CLs collected at different stages of the oestrous cycle. CLs were collected surgically, mRNA and protein extracted, and expression/levels analyzed by semi‐quantitative (SQ) PCR and Western blots, respectively. At the mRNA expression level, only BAX (maximal on day 4) and TP53 (maximal on day 7) showed significant variations during the oestrous cycle. At the protein level, only Bcl‐x and Caspase‐3 showed significant changes during the cycle; Bcl‐x decreased on day 13 and Caspase‐3 increased on day 13. It is concluded that apoptosis‐associated proteins (i.e. Bcl‐x and Caspase 3) may play a critical role in luteolytic sensitivity in the pig.     

Correlation between clinical signs of depth of anaesthesia and cerebral state index responses in dogs during induction of anaesthesia with propofol

The cerebral state index (CSI) is used for monitoring EEG and depth of anaesthesia. The objective of this study was to analyse the correlation between ocular reflexes, CSI and estimated propofol plasma concentrations (PropCP) in dogs during induction of anaesthesia with propofol.Fourteen dogs were premedicated with acepromazine 0.05 mg kg⁻¹ IM. Anaesthesia was induced with a 200 ml h⁻¹ propofol 1% constant infusion rate until loss of corneal reflex using RugLoop II software with Beths’ pharmacokinetic model to estimate PropCp.Palpebral reflex (PR) and the corneal reflex (CR) were tested every 30 s and classified as present (+) or absent (−), and eyeball position was registered as rotated ventromedialy (ERV) or centred (EC).Heart rate (HR), mean arterial pressure (MAP) and CSI values were analyzed from baseline before the beginning of propofol infusion (T0) until loss of CR; CSI and PropCp, CSI and anaesthetic planes, and PropCp and anaesthetic planes were compared using correlation analysis.PropCp reached 7.65 ± 2.1 μg ml⁻¹ at the end of the study. CSI values at T0 were 89.2 ± 3.8. Based on the observation of ocular reflexes and eyeball position, it was possible to define five anaesthetic planes: A (superficial) to E (deep), being A (PR+/CR+/EC), B (PR+/ERV/CR+), C (PR−/ERV/CR+), D (PR−/EC/CR+) and E (PR−/EC/CR−). There was a significant correlation between PropCp and the anaesthetic planes (R = 0,861; P < 0.01). No significant correlation was observed between CSI and the anaesthetic planes or between CSI and PropCp. MAP decreased significantly from T0 until loss of corneal reflex (from 98 ± 14 mmHg to 82 ± 12 mmHg); HR did not change significantly (from 101 ± 30 bpm to 113 ± 16 bpm).The CSI monitoring was not consistent with the clinical observations observed in the different stages of depth anaesthesia. This could limit the use of CSI for monitoring depth of anaesthesia with propofol.     

Effects of Plasma Urea Nitrogen Levels on the Bovine Oocyte Ability to Develop After In vitro Fertilization

The overall aim of the present study was to evaluate in vitro development ability of oocytes recovered from 56 Holstein Frisian heifers with low [group 1 (G1): <13 mg /dl], moderate [group 2 (G2): 13–16 mg /dl] and high [group 3 (G3): >16 mg /dl] plasma urea nitrogen (PUN) concentrations, to determine whether PUN concentrations affect the competence of oocytes to progress to blastocysts after in vitro fertilization. In vitro oocyte and embryo development was assessed by blastocyst rates, embryo total cell numbers and apoptosis. Blood samples for the determination of PUN were collected 24 h prior to collection of the ovaries at the slaughter. A total of 112 ovaries were collected at a local abattoir and oocytes (n = 697) were aspirated, in vitro matured and fertilized. On day 8, blastocysts were assigned to the terminal dUTP nick end labelling assay. Cleavage rates were significantly higher (p < 0.001) for groups 1 and 2 than for group 3 (i.e. 72.5% and 72.2% vs 61.7%, respectively). The proportion of fertilized oocytes that developed into blastocysts was higher (p < 0.05) for group 1 than for group 3 (34.0% vs 23.0%, respectively). Day 8 blastocysts showed higher total cell counts (p < 0.05) for group 1 than for group 3 (123.7 vs 76.3), and a higher (p < 0.05) total apoptotic cell rate was found in group 3 (25.9 and 19.0 vs 43.2 for G1, G2 and G3, respectively). In conclusion, the ability of oocytes from heifers with increased levels of PUN to develop to the blastocyst stage was significantly reduced when standard routines for in vitro maturation, fertilization and culture were followed. These detrimental effects can be mediated in part through direct effect of urea and/or by the metabolic products on the process of follicle-enclosed oocyte nuclear and cytoplasmic development.     

Phytotoxicity and genotoxicity of soils from an abandoned uranium mine area

The great number of abandoned uranium mines within the Portuguese territory led to the production of specific legislation establishing the rehabilitation of these areas and the definition of those requiring prior intervention. Cunha Baixa uranium mine (Mangualde, Central Portugal) was one of these areas and a site-specific risk assessment is being carried out for this mine, following European frameworks. In this study the Ames test with soil elutriates, and emergence and growth assays with Lactuca sativa L. and Zea mays L. were performed to assess soil retention and habitat functions. Levels of metals bioaccumulated in plant tissues, grown above soil surface, during the assays, were determined as well. No genotoxic effects were recorded for soil eluates (1:10 and 1:2, m/v) except for site F within the mine area, which is under the effluence of runoffs from the effluent treatment pond. Nevertheless, most of all were highly toxic to Salmonella typhimurium (especially for TA98) strains, used in the assay. The results gathered suggested that during the natural process of soil percolation by rainwater, the leachates yielded are contributing with toxic but no genotoxic metals to underground water resources. Phytotoxicity assays demonstrated that soil habitat function was compromised in almost all the sampling sites selected within the mine area. These results have clearly demonstrated how evaluations based on extractable metal concentrations may underestimate the risks of contaminated soils. Lettuce was the most sensitive species probably due to the great ability of this species in accumulating high concentrations of metals in their aboveground tissues. The highest bioaccumulation factors (BAFs) values were recorded for this species namely for Co (16.5), Cu (28.5), Ba (7.3), Sr (7.2) and Ni (6.1). Copper (9.0) and nickel (3.6) were the main elements bioaccumulated by maize. The dilution of mine soils with a local reference soil has proved to mitigate its toxicity in almost all the sites analysed.     

Acibenzolar-S-methyl doses and application methods to Pratylenchus brachyurus control in maize

Pratylenchus brachyurus is one of the main limiting factors of agricultural yield in annual crops, particularly due its difficult management. Methods to reduce populations are constantly sought, and the use of resistance inducers deserves attention. Thus, the study aimed to assess the effect of acibenzolar-S-methyl (ASM) application in different doses to P. brachyurus control in maize. Initially, 1000 specimens of P. brachyurus were inoculated in maize for the multiplication. After 60 days, the aerial part (AP) was discarded and maize was sown under the treatments: seed treatment (ST), application in the AP and ST?+?AP, in the doses of 0.125, 0.25, 0.5 and 1.0?g i.a./L. Vegetative and nematological parameters were assessed 60 days later. ASM did not affect plant growth, but reduced nematode population in all the treatments. The treatment methods did not differ and reduced the number of nematodes/g root by 64.25 to 80.14% compared to the control. The doses of 0.6, 0.51 and 0.65?g i.a./L promoted the most reductions in the number of nematodes/g root when applied in ST, AP and ST?+?AP, respectively. Thus, we recommend the use of ASM applied on shoot in a dose of 0.50?g i.a./L.     

Daylight effect on melatonin secretion in adult female guanacos (Lama guanicoe)

The wild South American camelids developed a strategy of seasonal reproduction during spring and summer with singleton birth. The photoperiod is one of the factors that may modulate this seasonality where light would be translated into a hormonal signal. This study evaluated the influence of changes in daily light intensity on melatonin concentration in captive guanacos under a long‐day photoperiod (16 hr light/8 hr dark; 33 ‘28′S). Mean melatonin concentration was 28.3 ± 20.3 pg/ml, with a maximum of 52.14 ± 17.19 pg/ml at 23:30 and minimum of 14.29 ± 6.64 pg/ml at 08:30 (p < .001). There was a negative association between light intensity and melatonin concentration (r = ?0.57; p < .001). The results indicate that guanacos respond to variation in daily environmental light with a hormonal response and point to a circannual rhythm as a function of the photoperiod.     

Effects of different propofol infusion rates on EEG activity and AEP responses in rats

Parameters calculated from the auditory-evoked potential (AEP) recorded over the auditory cortex and from the electroencephalogram (EEG) recorded over the near vertex were compared in rats at three different infusion rates of propofol (62.5, 35 and 25 mg/kg/h). Depth of anaesthesia was assessed clinically using the strength of the pedal withdrawal reflex. Well-defined AEP responses were consistently obtained. As the propofol concentration was reduced, peak latencies decreased and peak to peak amplitudes increased. Amplitude and latency values were closely associated with the strength of the pedal withdrawal responses. Parameters calculated from the EEG showed no significant change as the propofol concentration was reduced. Periods of burst suppression became more frequent as the propofol infusion rate was increased. The study showed some of the difficulties that may be encountered when using EEG as a tool to assess depth of anaesthesia during propofol infusion. The AEP showed dose dependent changes in rats at different infusion rates of propofol. However, large variability between animals limits the use of this technique for monitoring depth of anaesthesia.