Impacts of rumen fluid, refrigerated or reconstituted from a refrigerated pellet, on gas production measured at 24h of fermentation

3noRumen fluid is used as fresh inoculum for gas production fermentations to predict the nutritional value of feeds and rations for ruminants. However, collection of rumen fluid from animal donors is invasive, expensive, time consuming and results in fluids of variable quality. The general aim was to identify a procedure to manipulate rumen inoculum in order to facilitate its storage and transfer between laboratories. This strategy would also limit fluid collections from animals. Two experiments were completed based on gas production from graduated 100 mL glass syringe with five feeds as substrates. In experiment 1, the gas production and some fermentation parameters of fresh rumen fluids were compared with those preserved at 4 °C for 24, 48, 72 and 96 h. Refrigeration did not modify concentration of volatile fatty acids and pH, but ammonia in liquids refrigerated for 48–96 h was higher (P < 0.05) compared to fresh. In contrast, rumen fluid refrigeration for 24, 48 or 72 h did not depress gas production at 24 h, but it was lower at 96 h. In experiment 2, the rumen fluid was centrifugated at 13,000 x g and sedimented material (i.e., pellet) was refrigerated for 48 h at 4 °C. The asymptote of gas production kinetics from rumen fluid regenerated from the pellet was 8 % lower (P < 0.05) than that from fresh. However for 24 h gas production, the correlation between fresh liquid and pellet inoculum, calculated for five ingredients, was high (R2 = 0.94). Results support the use of rumen fluid preserved by refrigeration for up to 72 h, and rumen fluid reconstituted from refrigerated pellet, as an alternative to fresh. This would reduce the need for laboratories to maintain animal donors and/or frequently collect rumen fluid.openopenFabro C.; Sarnataro C.; Spanghero M.Fabro, C.; Sarnataro, C.; Spanghero, M

In vitro ammonia release of urea-treated high moisture barley and maize grain

Rumen nitrogen (N) release from ammoniated wet barley and maize kernels by urea treatment (UT) at harvesting was studied. Untreated samples (CTR) were compared to UT and to samples combined with urea just before the experiment (UA). In Experiment 1, ground CTR, UT and UA samples were fermented in a ruminal in vitro system, and ammonia of fermentation fluid was analysed at 0, 2, 4, 6 and 8 h. The effect of incubation time was observed as ammonia peaked at 4 h of fermentation (10.24 vs 9.01 and 7.20 mg \ub7 dl 121, respectively at 0 and 8 h, P < 0.01). Also, the effect of treatment was stated when UT released less ammonia than UA treatment (9.76 vs 10.52 mg \ub7 dl 121, P < 0.05), while the CTR samples showed the least ammonia N concentrations (P < 0.01). In Experiment 2, the water N solubility of CTR and UT of both cereal samples prepared in three physical forms (whole grain, coarsely ground and milled) was examined. Samples were incubated in flasks with distilled water for 1, 2, 4, 6 and 8 h and N was measured in filtered residues to calculate N solubility. The UT samples, regardless cereal type, solubilised more N in the milled than in the whole form with the coarse form in the middle (43.7 vs 15.3%, 32.4 vs 14.0% and 20.3 vs 9.2% for milled, coarse and whole form, respectively; treatment 7 physical form interaction: P < 0.01). The N added to wet cereal kernels by the urea treatment was released in the rumen fermentation liquid more slowly than that simply added as urea before incubation. Based on solubility data, the treated whole or cracked kernels exhibited a slower N release than milled ones

Effect of dietary nitrogen level and source on mRNA expression of urea transporters in the rumen epithelium of fattening bulls

This paper aims to study the effect of the dietary treatments on mRNA expression of urea transporter B (UT-B) and some aquaporins (AQP) in rumen epithelium of Italian Simmental young bulls. Eighty animals allocated to 16 pens were fed from about 500 to 650&nbsp;kg body weight with four experimental diets, which resulted from the combination of two crude protein levels (125 and 110&nbsp;g/kg dry matter, diets M and L, respectively) and two nitrogen sources (soybean meal (SBM) or SBM partly replaced by an isonitrogenous mixture of corn and urea; diets −U and +U, respectively). At slaughtering samples of blood and rumen epithelium were collected from six bulls for each diet. Blood samples were analysed for haematological parameters and quantitative PCR was carried out on the mRNA extracted from the rumen epithelium samples. The bulls fed diets M had lower plasma concentrations of aspartate aminotransferase than those receiving diets L (78.9 vs. 88.3&nbsp;U/l, p&nbsp;=&nbsp;0.04). Plasma urea was higher (p&nbsp;=&nbsp;0.03) for diets M and lower for diets +U (2.0 vs. 2.5 and 1.73 vs. 2.00&nbsp;mmol/l, respectively, in M and L diets, p&nbsp;=&nbsp;0.04). The effect of dietary treatments on rumen UT expression were limited to AQP3, which was down regulated (p&nbsp;=&nbsp;0.01) in diets +U. Finally, a high positive correlation (R2&nbsp;=&nbsp;0.871) between the expressions of AQP7 and AQP10 was found. In conclusion, the AQP3 appears very responsive to dietary treatments and therefore it is a candidate to be further studied in rumen metabolism experiments. The close relationship between mRNA expression of AQP7 and AQP10 indicates a similar function of these two proteins

In vitro aflatoxins recovery after changing buffer or protozoa concentrations in the rumen fermentation fluid

This study simulates in vitro the effects of (i) rumen acidity and (ii) change in rumen protozoa numbers on the recovery of aflatoxins (AFs). Two 24-h fermentation experiments were carried out using the same batch in vitro fermentation systems and substrate (dried corn meal) containing 11.42, 2.42, 7.65 and 1.70 µg/kg of AFB1, AFB2, AFG1 and AFG2 respectively. In Experiment 1, two buffer concentrations (normal salts dosage or lowered to 25%) were tested. Buffer reduction decreased gas production (730 vs. 1101 mL, p < 0.05), volatile fatty acids (VFA) and NH3 concentrations in the fermentation liquid (39.8 vs. 46.3 mmol/L, and 31.7 vs. 46.5 mg/dL respectively, p < 0.01). Recovery of all four AFs types was higher (p < 0.01) in the reduced buffer fermentation fluid, both as a percentage of total AF incubated (73.6% vs. 62.5%, 45.9% vs. 38.1%, 33.6% vs. 17.9% and 18.9% vs. 6.24% for AFB1, AFB2, AFG1 and AFG2 respectively) and as amounts relative to VFA production (163.4 vs. 123.5, 22.1 vs. 15.7, 48.8 vs. 22.5 and 6.16 vs. 1.86 ng/100 mmol of VFA, for AFB1, AFB2, AFG1 and AFG2 respectively). In Experiment 2, Stevia rebaudiana Bertoni extracts (S) or a Camphor essential oil (Cam) were added to fermenters and compared to the control (no additives, C). S and Cam addition resulted in a 25% reduction (p < 0.05) and a 15% increase (p < 0.05) in protozoa counts respectively, when compared to C. Both plant additives slightly reduced (p < 0.05) AFB1 recovery as a percentage of total AFB1 incubated (68.5% and 67.7% vs. 74.9% for S, Cam and C respectively). Recoveries of all other AFs were unaffected by the additives. In conclusion, the rumen in vitro AFB1 recovery (63%–75%) was higher than other AFs (3%–46%) and the acidic fermentation environment increased it. In our conditions, changes in protozoa numbers did not affect AFs recovery

The Design Development of Solar Box Cooker on Wheels as an Alternative for Philippine Rural Households

Cooking accounts for a significant share of the worldwide energy consumption issue. Solar box cookers are a good alternative in mitigating fossil fuels and biomass use, especially for tropical developing nations like the Philippines. Still, their broad acceptance is limited by their high dependence on sufficient sunlight for better performance. Thus, this research aims to develop a thermal-efficient, portable solar box cooker with sand-sensible heat storage materials, coconut coir (Cocos nucifera) agricultural waste insulators, and locally sourced building materials. The designed and constructed cooker was evaluated using three thermal performance tests: the Stagnation Temperature test from the Bureau of Indian Standards (BIS), the Cooking Power test from the American Societal of Agricultural Engineer (ASAE) Standards, and a rice cooking test. Data obtained from the three-day trials of each test were used to determine the first figure of merit (F1), the single measure of performance (Ps(50)), and its ability to cook 300 g of NFA rice with 500 mL of water, respectively. It was found that the cooker achieved an F1 of 0.06 Km2/w, a Ps(50) of 6.651 W, and cooked rice for one-third of the tests. Obtained experimental findings showed that the device is marked as a Grade-B solar cooker. Results indicated that the solar box cooker is capable of pasteurizing water and cooking rice

Heat stress and feeding behaviour of dairy cows in late lactation

Heat stress is one of the most important problems that dairy cows have to face and the use of cooling systems is becoming more and more important. The first reaction that has the animal to cope with the environmental variations is to modify its behaviour. This study was aimed to investigate the effect of heat stress and a cooling system on the feeding behaviour of Italian Holstein Friesian dairy cows in late lactation. Two experiments were performed. In the first experiment, eight dairy cows were firstly kept 7 d under thermoneutral condition, and then under mild heat stress (temperature humidity index, THI, ranging between 72 and 78) for others 7 d. The second experiment consisted of 8 dairy cows used in a two-period cross-over design where the treatment was the use or not of a sprinkler system for cooling cows under mild heat stress. Cows were equipped with a noseband pressure sensor able to detect rumination and eating time, number of rumination and eating chews, number of rumination boluses and rumination intensity. Heat stress reduced rumination time, number of rumination chews and boluses (p <.05), and tended to reduce the number of eating chews (p <.10). Cooled cows increased rumination and eating time (p <.05), rumination intensity (p <.01), and the number of rumination and eating chews (p <.05). In conclusion, feeding behaviour was deeply influenced even by mild heat stress, which was effectively improved by the use of a sprinkler system.HIGHLIGHTS Mild heat stress reduced rumination time, number of rumination chews and boluses of dairy cows in late lactation Cooling cows with sprinklers was effective in alleviating heat stress in terms of feeding behaviour

Trend of decreasing length of cervical cone excision during the last 20 years

OBJECTIVE: The aim of the present investigation was to evaluate the cervical conizations performed in the last 20 years in a single institution, with a particular interest in analyzing the trend of the length of cone excisions.PATIENTS AND METHODS: A retrospective cohort study of women who underwent a CO2-laser cervical conization between January 1996 and December 2015. Cytological abnormalities on referral pap smear, colposcopic findings and pertinent clinical and socio-demographic characteristics of each woman were collected. In particular, the length of cone specimen was evaluated, taking into account all the factors potentially influencing the length of excision.RESULTS: A total of 1270 women who underwent cervical conization from January 1996 to December 2015 were included in the analysis. A mean cone length of 15.1 \ub1 5.7 mm was reported, and we observed a significant decrease in the length of cone excisions over the whole study period. Age (rpartial = 0.1543, p < 0.0001), see & treat procedure (rpartial = -0.1945, p < 0.0001) and grade II colposcopic findings (rpartial = 0.1540, p < 0.0001) were significantly associated with the length of cone excision on multivariate analysis.CONCLUSIONS: In the last 20 years, a significant decrease in the length of cone excision was observed. In our opinion, this can be due to the acquired awareness by the gynecologists of the potential disadvantages of wide cone excision in term of adverse obstetric outcomes in future pregnancies

Phospholipase C2 Affects MAMP-Triggered Immunity by Modulating ROS Production

The activation of phosphoinositide-specific phospholipase C (PI-PLC) is one of the earliest responses triggered by the recognition of several microbe-associated molecular patterns (MAMPs) in plants. The Arabidopsis (Arabidopsis thaliana) PI-PLC gene family is composed of nine members. Previous studies suggested a role for PLC2 in MAMP-triggered immunity, as it is rapidly phosphorylated in vivo upon treatment with the bacterial MAMP flg22. Here, we analyzed the role of PLC2 in plant immunity using an artificial microRNA to silence PLC2 expression in Arabidopsis. We found that PLC2-silenced plants are more susceptible to the type III secretion system-deficient bacterial strain Pseudomonas syringae pv tomato (Pst) DC3000 hrcC− and to the nonadapted pea (Pisum sativum) powdery mildew Erysiphe pisi. However, PLC2-silenced plants display normal susceptibility to virulent (Pst DC3000) and avirulent (Pst DC3000 AvrRPM1) P. syringae strains, conserving typical hypersensitive response features. In response to flg22, PLC2-silenced plants maintain wild-type mitogen-activated protein kinase activation and PHI1, WRKY33, and FRK1 immune marker gene expression but have reduced reactive oxygen species (ROS)-dependent responses such as callose deposition and stomatal closure. Accordingly, the generation of ROS upon flg22 treatment is compromised in the PLC2-defficient plants, suggesting an effect of PLC2 in a branch of MAMP-triggered immunity and nonhost resistance that involves early ROS-regulated processes. Consistently, PLC2 associates with the NADPH oxidase RBOHD, suggesting its potential regulation by PLC2

Production and evaluation of leukocyte- and thrombocyte-rich fibrin membranes in birds

The aim of this study was the preparation and histological evaluation of Leukocyte- and Thrombocyte-Rich Fibrin (L-TRF) membranes obtained from the blood of four bird species. Forty adult healthy birds were divided into four groups of equal size: G1 – macaws, G2 – domestic chickens, G3 – parrots, G4 – toco toucans. A total of 0.5 mL of blood was collected from each bird, put into a glass tube without anticoagulant and centrifuged at 3000 rpm for 10 min. L-TRF membranes produced after compression of the clot were processed for histological analysis. The ratio of thrombocytes/area was not significantly different among Groups G2, G3 and G4, but a significant difference was found between Groups G1 and G2 with the highest thrombocyte concentration/area in G1. The groups did not differ statistically in the number of leukocytes/area. The fibrin-to-cells ratio did not vary statistically among Groups G1, G2 and G3, but this ratio was significantly higher in Group G4 than in the other groups. The thrombocyte-to-leukocyte ratio was the highest in Group G1, but it did not differ among Groups G2, G3 and G4. In conclusion, the centrifugation protocol allowed the production of L-TRF membranes in the four bird species studied. Histologically, cell ratios were analogous in domestic chickens and parrots, and macaws had the highest ratio of thrombocytes