Chitosan nanoparticles enhance developmental competence of in vitro-matured porcine oocytes

Oxidative stress is inevitable as it is derived from the handling, culturing, inherent metabolic activities and medium supplementation of embryos. This study was performed to investigate the protective effect of chitosan nanoparticles (CNPs) on oxidative damage in porcine oocytes. For this purpose, cumulus–oocyte complexes (COCs) derived from porcine slaughterhouse ovaries were exposed to different concentrations of CNPs (0, 10, 25 and 50 µg/ml) during in vitro maturation (IVM). Oocytes treated with 25 µg/ml CNPs showed significantly higher levels of GSH, along with a significant reduction in ROS levels compared to control, CNPs10 and CNPs50 groups. In parthenogenetic embryo production, the maturation rate was significantly higher in the CNPs25 group than that in the control and all other treated groups. In addition, when compared to the CNPs50 and control groups, CNPs25-treated oocytes showed significantly higher cleavage and blastocyst development rates. The highest concentration of CNPs reduced the total cell number and ratio of ICM: TE cells in parthenogenetic embryos, suggesting that there is a threshold where benefits are lost if exceeded. In cloned embryos, the CNPs25 group, as compared to all other treated groups, showed significantly higher maturation and cleavage rates. Furthermore, the blastocyst development rate in the CNPs25-treated group was significantly higher than that in the CNPs50-treated group, as was the total cell number. Moreover, we found that cloned embryos derived from the CNPs25-treated group showed significantly higher expression levels of Pou5f1, Dppa2, and Ndp52il genes, compared with those of the control and other treated groups. Our results demonstrated that 25 µg/ml CNPs treatment during IVM improves the developmental competence of porcine oocytes by reducing oxidative stress.     

Rapidly quantitative detection of Nosema ceranae in honeybees using ultra-rapid real-time quantitative PCR

BackgroundThe microsporidian parasite Nosema ceranae is a global problem in honeybee populations and is known to cause winter mortality. A sensitive and rapid tool for stable quantitative detection is necessary to establish further research related to the diagnosis, prevention, and treatment of this pathogen.ObjectivesThe present study aimed to develop a quantitative method that incorporates ultra-rapid real-time quantitative polymerase chain reaction (UR-qPCR) for the rapid enumeration of N. ceranae in infected bees.MethodsA procedure for UR-qPCR detection of N. ceranae was developed, and the advantages of molecular detection were evaluated in comparison with microscopic enumeration.ResultsUR-qPCR was more sensitive than microscopic enumeration for detecting two copies of N. ceranae DNA and 24 spores per bee. Meanwhile, the limit of detection by microscopy was 2.40 × 10⁴ spores/bee, and the stable detection level was ≥ 2.40 × 10⁵ spores/bee. The results of N. ceranae calculations from the infected honeybees and purified spores by UR-qPCR showed that the DNA copy number was approximately 8-fold higher than the spore count. Additionally, honeybees infected with N. ceranae with 2.74 × 10⁴ copies of N. ceranae DNA were incapable of detection by microscopy. The results of quantitative analysis using UR-qPCR were accomplished within 20 min.ConclusionsUR-qPCR is expected to be the most rapid molecular method for Nosema detection and has been developed for diagnosing nosemosis at low levels of infection.     

The effects of maxillary nerve block,ethmoidal nerve block and their combination on cardiopulmonary responses to nasal stimulation in anesthetized Beagle dogs

ObjectiveTo describe an approach for ethmoidal nerve block (E_BLOCK) and to compare the effects of a maxillary nerve block (M_BLOCK), E_BLOCK and their combination (M-E_BLOCK) on heart rate (HR), systolic (SAP), mean (MAP), diastolic (DAP) arterial pressures and respiratory rate (f_R) during nasal stimulation in dogs.Study designProspective, blinded, randomized, crossover placebo-controlled study.AnimalsBeagle dogs (five cadavers, nine live dogs), with a median (interquartile range) weight of 10.5 (10.3–11.0) kg.MethodsThe accuracy of iohexol injections (each 1 mL) at the maxillary and ethmoidal foramina in cadavers was evaluated using computed tomography. Then, anesthetized dogs were administered four bilateral treatments separated by 1 week, saline or 2% lidocaine 1 mL per injection: injections of saline at the maxillary and ethmoidal foramina (Control), injections of lidocaine at the maxillary foramina and saline at the ethmoidal foramina (M_BLOCK), injections of saline at the maxillary foramina and lidocaine at the ethmoidal foramina (E_BLOCK) and injections of lidocaine at all foramina (M-E_BLOCK). The ventral nasal meatus was bilaterally stimulated using cotton swabs, and HR, SAP, MAP, DAP and f_R were continuously recorded. Values for each variable were compared before and after stimulation using Wilcoxon signed-rank test. Changes in variables among treatments were analyzed using Mann–Whitney U and Kruskal–Wallis tests (p ≤ 0.05).ResultsComputed tomography revealed iohexol distribution around the openings of the target foramina in all cadavers. In living dogs, HR, SAP, MAP, DAP and f_R significantly increased after stimulation within each treatment (p < 0.03). Physiologic responses were significantly attenuated, but not absent, in the M-E_BLOCK [HR (p = 0.019), SAP, MAP, DAP and f_R (all p ≤ 0.001)] compared with those in the Control.Conclusions and clinical relevanceConcurrent injections of lidocaine at the maxillary and ethmoidal foramina attenuated HR, arterial pressure and f_R responses to nasal stimulation in Beagle dogs.     

Association between weight change during initial chemotherapy and clinical outcome in dogs with multicentric lymphoma

The majority of the known prognostic factors in dogs with lymphoma have been evaluated before treatment commences or at the time of diagnosis. Prognostic factors evaluated during the initial phase of treatment are less described but may provide important clinical information. In this retrospective study, 82 canine lymphoma patients were categorized according to the weight change between diagnosis and after 5 weeks of chemotherapy. Dogs that gained greater than 5% or lost greater than 5% of initial body weight were categorized as increased‐ or decreased‐weight groups, respectively. Those in which weight changed less than 5% were categorized as the maintained‐weight group. The median progression‐free survival (PFS) in the increased‐weight group, maintained‐weight group and decreased‐weight group was 226, 256 and 129 days, respectively. The decreased‐weight group had significantly shorter PFS than the increased and maintained groups (P = .023, P = .003, respectively). The median survival time (ST) in the increased‐weight group, maintained‐weight group and decreased‐weight group was 320, 339 and 222 days, respectively. There was no significant difference in ST among the three groups (P = .128). In Cox‐regression results, weight change group and initial body weight were significant risk factors associated to PFS (P = .007, P = .001, respectively) while only patient's initial body weight was a significant risk factor to ST (P = .013). In conclusion, evaluation of initial body weight and weight changes over time can provide valuable information regarding PFS and ST in dogs with multicentric lymphoma.     

Effects of dietary supplementation of nucleotides from late gestation to lactation on the performance and oxidative stress status of sows and their offspring

Increased metabolic burdens in breeding sows, which are induced by elevated systemic oxidative stress, could increase the need for nucleotides to repair lymphocyte DNA damage; however, de novo synthesis of nucleotides may be insufficient to cover this increased need. This study investigated the effects of dietary nucleotides on milk composition, oxidative stress status, and the reproductive and lactational performance of sows. Forty multiparous sows were assigned to 2 dietary treatments (Control group, and 1 g/kg Nucleotides group) based on a randomized complete block design using their BW at 85 d of gestation as a block. Sows from 2 groups were fed a restricted diet during gestation and ad libitum during lactation. The experiment lasted from 85 d of gestation to 21 d of lactation. The reproductive performance of sows and the growth performance of suckling piglets were measured. Oxidative stress parameters and milk components were also analysed. Data were analyzed using contrasts in the MIXED procedure of SAS. Sows in the Nucleotides group consumed more feed during the first week (P < 0.01) and from 1 to 21 d (P < 0.05) of lactation than those in Control group. Correspondingly, the litter weight gain of piglets showed a tendency to increase from cross-fostering to 9 d (P = 0.09) and from cross-fostering to 20 d (P = 0.10) in the Nucleotides group relative to the Control group. Additionally, the Nucleotides group was higher (P < 0.01) than the Control group in the concentrations of uridine 5''monophosphate, guanosine 5''monophosphate, inosine 5''monophosphate, adenosine 5''monophosphate and total nucleotides in milk. Furthermore, the Nucleotides group was higher (P < 0.01) than the Control group in the serum levels of total antioxidant capacity (P < 0.01) for sows at 109 d of gestation and glutathione peroxidase for weaning piglets, but lower at the levels of thiobarbituric acid-reactive substances (P < 0.05) in serum of weaning piglets. This study indicated that maternal dietary nucleotides could promote piglet growth, probably due to the higher lactational feed intake and higher concentration of nucleotides in the milk of sows, and lower oxidative stress for both sows and piglets.     

Protein and energy concentrations of meat meal and meat and bone meal fed to pigs based on in vitro assays

The objective of this study was to develop equations for estimating ileal digestible crude protein (CP) and metabolizable energy (ME) contents of meat meal (MM) and meat and bone meal (MBM) as feed ingredients for pigs based on in vitro assays. Test ingredients were 4 sources of MM and 3 sources of MBM. Ash and CP contents of the ingredients ranged from 3.8% to 33.1% and 46.8% to 82.9% (as-is basis), respectively. In vitro ileal disappearance (IVID) of CP was determined and ileal digestible CP content was calculated by multiplying CP content by IVID of CP. In vitro total tract disappearance (IVTTD) of dry matter (DM) was determined and ME was calculated using gross energy, CP contents, and IVTTD of DM. The IVID of CP and IVTTD of DM ranged from 77.2% to 88.7% and from 82.7% to 92.4%, respectively. Calculated ileal digestible CP and ME contents ranged from 37.8% to 73.5% DM and 2,405 to 3,905 kcal/kg DM, respectively. Ash contents were negatively correlated (P < 0.001) with CP (r = −0.99), in vitro ileal digestible CP (r = −0.97), gross energy (r = −1.00), in vitro digestible energy (r = −0.97), and adjusted ME (r = −0.97). The most fitting equations for ileal digestible CP and adjusted ME were: ileal digestible CP (% DM) = 11.91 − 0.90 × Ash (% DM) + 0.74 × IVID of CP (%) (R² = 0.99) and adjusted ME (kcal/kg DM) = 130.85 − 50.90 × ash (% DM) + 47.06 × IVTTD of DM (%) (R² = 0.99). To validate the accuracy of the prediction equations for ME, mean bias and linear bias were determined using a regression analysis. Calculated ME values of MM and MBM were in a good agreement with data obtained from animal experiments based on a statistically insignificant bias in the models. In conclusion, ME concentrations of MM and MBM as swine feed ingredients can be calculated using ash concentration and in vitro disappearance of dry matter.     

Heavy Metal Immobilization in Contaminated Soils using Phosphogypsum and Rice Straw Compost

The application of soil amendments to immobilize heavy metals is a promising technology to meet the requirements for environmentally sound and cost‐effective remediation. The present study was carried out to evaluate the result of phosphogypsum (PG) used alone and in combination with compost (CP) at a mix ratio of 1:1 wet weight ratio (PG + CP) at 10 and 20 g dry weight kg⁻¹ dry soil, on heavy metal immobilization in contaminated soil and on canola growth. The results revealed that the Pb, Cd and Zn uptake of canola plants was reduced by the application of PG alone and when it was mixed with CP as compared with untreated soil. At an application rate of 10 g dry weight kg⁻¹ dry soil of (PG + CP) the dry weight of canola plants increased by 66·8% was increased in comparison with its weight in the untreated soil. The addition of PG alone resulted in more pronounced immobilization of heavy metal as compared with PG mixed with CP. Plant growth was improved with CP addition, but heavy metals immobilization was the greatest in PG alone treatments. Results suggest that PG may be useful for the immobilization of heavy metals in contaminated soils. Copyright © 2014 John Wiley & Sons, Ltd.     

Impact of Land Use and Land Cover Changes on Organic Carbon Stocks in Mediterranean Soils (1956–2007)

During the last few decades, land use changes have largely affected the global warming process through emissions of CO₂. However, C sequestration in terrestrial ecosystems could contribute to the decrease of atmospheric CO₂ rates. Although Mediterranean areas show a high potential for C sequestration, only a few studies have been carried out in these systems. In this study, we propose a methodology to assess the impact of land use and land cover change dynamics on soil organic C stocks at different depths. Soil C sequestration rates are provided for different land cover changes and soil types in Andalusia (southern Spain). Our research is based on the analysis of detailed soil databases containing data from 1357 soil profiles, the Soil Map of Andalusia and the Land Use and Land Cover Map of Andalusia. Land use and land cover changes between 1956 and 2007 implied soil organic C losses in all soil groups, resulting in a total loss of 16·8 Tg (approximately 0·33 Tg y⁻¹). Afforestation increased soil organic C mostly in the topsoil, and forest contributed to sequestration of 8·62 Mg ha⁻¹ of soil organic C (25·4 per cent). Deforestation processes implied important C losses, particularly in Cambisols, Luvisols and Vertisols. The information generated in this study will be a useful basis for designing management strategies for stabilizing the increasing atmospheric CO₂ concentrations by preservation of C stocks and C sequestration. Copyright © 2012 John Wiley & Sons, Ltd.     

Effect of the Nixtamalization Process on the Dietary Fiber Content,Starch Digestibility,and Antioxidant Capacity of Blue Maize Tortilla

Nixtamalization is an ancient process developed by the Mesoamerican cultures. Initially, volcanic ashes were used and then calcium hydroxide in commercial production, and more recently nixtamalization with calcium salts (NCS) has been proposed. The aim of this study was to evaluate the effect of NCS on carbohydrate digestibility and antioxidant capacity in the elaboration of blue maize tortillas. NCS in blue tortillas showed a high amount of total dietary fiber (14.27 g/100 g), the main fraction being insoluble dietary fiber. The contents of resistant starch and slowly digestible starch did not change with the nixtamalization process. The predicted glycemic index value was lower in blue tortillas with the NCS process (58) than with the traditional nixtamalization process (71). In general, NCS in blue tortillas presented a higher antioxidant capacity than traditional tortillas (ferric reducing antioxidant power method), indicating that phenolics present in blue maize maintain their activity after cooking. It can be concluded that the nutraceutical features (high dietary fiber content and antioxidant capacity) of blue maize tortillas are enhanced when they are elaborated with the NCS process.