Valuation of angiogenesis in bovine xenografts implanted in intracorporal sites of rabbits as models of in-vivo bioreactors / Avaliação da angiogênese em xenoenxertos bovinos implantados em sítios intracorpóreos de coelhos como modelos de biorreatores in vivo

The aim of this study was to evaluate neovascularization of bovine xenografts implanted in intracorporeal sites of rabbits (bioreactors). 30 rabbits were used, divided into 6 groups, according to the evaluation time (7, 15, 30, 45, and 60 days); each animal received xenogenic implants in 3 different intracorporeal sites (A1 - omentum bag; A2 - intermuscular space of quadriceps femoris; A3 - subperiosteal of ilium bone). Histological assessments graded the presence of angiogenesis, the number of inflammatory cells, newly formed bone tissue, and the presence of giant cells. Histological analyses showed intense angiogenesis in all implanted xenografts. Presence of inflammatory infiltrate and giant cells at the A1 implant site and presence of bone neoformation at the A3 implant site were noted. Degeneration of implants and formation of a fibrous capsule were noted. When comparing the interaction of the site with the days of evaluation, statistical analysis showed a significant difference (p≤0.05) in any time of neovascularization analysis. The vascular endothelial growth factor (VEGF) and inflammatory cells of the omentum in its structure, may have contributed to the greater presence of neovessels and inflammatory cells, a fact that may indicate functionality as a possible bone substitute.(AU)

O objetivo deste estudo foi avaliar a neovascularização de xenoenxertos bovinos implantados em sítios intracorpóreos de coelhos (biorreatores). Foram utilizados 30 coelhos, os quais foram divididos em seis grupos, de acordo com o tempo de avaliação (sete, 15, 30, 45 e 60 dias); cada animal recebeu implantes xenogênicos em três diferentes sítios intracorpóreos (A1 - bolsa de omento; A2 - espaço intermuscular do quadríceps femoral; A3 - subperiosteal do osso ílio). Avaliações histológicas classificaram a presença de angiogênese, o número de células inflamatórias, de tecido ósseo neoformado e a presença de células gigantes. As análises histológicas mostraram intensa angiogênese em todos os xenoenxertos implantados. Observou-se presença de infiltrado inflamatório e células gigantes no local do implante A1 e presença de neoformação óssea no local do implante A3. Ao mesmo tempo, a degeneração dos implantes e a formação de uma cápsula fibrosa foram observadas. Ao comparar a interação do local com os dias de avaliação, a análise estatística mostrou diferença significativa (P≤0,05) em qualquer momento da análise de neovascularização. O fator de crescimento endotelial vascular (VEGF) e as células inflamatórias do omento em sua estrutura podem ter contribuído para a maior presença de neovasos e células inflamatórias, fato que pode indicar funcionalidade como possível substituto ósseo.(AU)

Copper sulfate and sodium selenite lipid-microencapsulation modifies ruminal microbial fermentation in a dual-flow continuous-culture system.

Undesirable interactions between trace mineral elements and ruminal contents may occur during digestion when mineral salts are supplemented. Antimicrobial effects of copper sulfate (CuSO4) may affect ruminal digestibility of nutrients when fed as a source of copper (Cu), while sodium selenite (Na2SeO3) may be reduced in the rumen to less available forms of selenium (Se). Our objective was to evaluate if protection of CuSO4 and Na2SeO3 by lipid-microencapsulation would induce changes on ruminal microbial fermentation. We used 8 fermentors in a dual-flow continuous-culture system in a 4 × 4 duplicated Latin square with a 2 × 2 factorial arrangement of treatments. Factors were CuSO4 protection (unprotected and protected by lipid-microencapsulation) and Na2SeO3 protection (unprotected and protected by lipid-microencapsulation). Treatments consisted of supplementation with 15 mg/kg of Cu and 0.3 mg/kg of Se from either unprotected or protected (lipid-microencapsulated) sources, as follows: (1) Control (unprotected CuSO4 + unprotected Na2SeO3); (2) Cu-P (protected CuSO4 + unprotected Na2SeO3); (3) Se-P (unprotected CuSO4 + protected Na2SeO3); (4) (Cu+Se)-P (protected CuSO4 + protected Na2SeO3). All diets had the same nutrient composition and fermentors were fed 106 g of dry matter/d. Each experimental period was 10 d (7 d of adaptation and 3 d for sample collections). Daily pooled samples of effluents were analyzed for pH, NH3-N, nutrient digestibility, and flows (g/d) of total N, NH3-N, nonammonia N (NAN), bacterial N, dietary N, and bacterial efficiency. Kinetics of volatile fatty acids was analyzed in samples collected daily at 0, 1, 2, 4, 6, and 8 h after feeding. Main effects of Cu protection, Se protection, and their interaction were tested for all response variables. Kinetics data were analyzed as repeated measures. Protection of Cu decreased acetate molar proportion, increased butyrate proportion, and tended to decrease acetate:propionate ratio in samples of kinetics, but did not modify nutrient digestibility. Protection of Se tended to decrease NH3-N concentration, NH3-N flow, and CP digestibility; and to increase flows of nonammonia N and dietary N. Our results indicate that protection of CuSO4 may increase butyrate concentration at expenses of acetate, while protection of Na2SeO3 tended to reduce ruminal degradation of N. Further research is needed to determine the effects of lipid-microencapsulation on intestinal absorption, tissue distribution of Cu and Se, and animal performance.

Ruminal fermentation and enteric methane production of legumes containing condensed tannins fed in continuous culture.

A continuous-culture fermentor study was conducted to assess nutrient digestibilities, volatile fatty acid (VFA) concentrations, microbial protein synthesis, bacterial nitrogen (N) efficiency, and enteric methane (CH4) production of four 50:50 grass-legume diets, randomly assigned in a 4 × 4 Latin square design. Four legumes with different concentrations of condensed tannins (CT) were tested: alfalfa [ALF; Medicago sativa L., non-CT legume]; birdsfoot trefoil [BFT; Lotus corniculatus L., low-CT legume]; crown vetch [CV; Securigera varia (L.) Lassen, moderate-CT legume]; and sericea lespedeza [SL; Lespedeza cuneata (Dum. Cours.) G. Don, high-CT legume]. Orchardgrass (Dactylis glomerata L.) was the common forage used in all diets. Four fermentors were evaluated over four 10-d periods by feeding 82 g of dry matter (DM)/d in 4 equal feedings. Methane output was recorded every 10 min. Effluent samples were collected during the last 3 d of the experiment, composited by fermentor and period, and analyzed for pH and VFA, as well as DM, organic matter, crude protein, neutral detergent fiber, and acid detergent fiber for determination of apparent and true nutrient digestibilities. Microbial protein synthesis and bacterial efficiency were estimated by analysis of N flows and purines. The CT concentrations were 3, 21, 38 and 76 g/kg of DM for ALF, BFT, CV, and SL diets, respectively. The SL diet had decreased fiber digestibilities and total VFA concentrations compared with the other diets. This resulted in the least total CH4 production in the SL diet. Bacterial N efficiency per kilogram of organic matter truly digested was lower in the SL diet than in the BFT and CV diets. The lowest CH4 production per unit of digestible nutrients was also found in the SL diet. Further work should be conducted to find optimal diets (by testing other legumes, rations, and sources of CT) for reducing CH4 emissions without negatively affecting ruminal digestion to maintain or improve productivity.

Diet starch concentration and starch fermentability affect markers of inflammatory response and oxidant status in dairy cows during the early postpartum period.

Our objective was to evaluate the effects of diet starch concentration and starch fermentability on inflammatory response markers and oxidant status during the early postpartum (PP) period and its carryover effects. Fifty-two multiparous Holstein cows were used in a completely randomized block design experiment with a 2 × 2 factorial arrangement of treatments. Treatments were starch concentration and starch fermentability of diets; diets were formulated to 22% (low starch, LS) or 28% (high starch, HS) starch with dry-ground corn (DGC) or high-moisture corn (HMC) as the primary starch source. Treatments were fed from 1 to 23 d PP and then switched to a common diet until 72 d PP to measure carryover (CO) effects. Treatment period (TP) diets were formulated to 22% forage neutral detergent fiber and 17% crude protein. The diet for the CO period was formulated to 20% forage neutral detergent fiber, 17% crude protein, and 29% starch. Coccygeal blood was collected once a week during the TP and every second week during the CO period. Liver and adipose tissue biopsies were performed within 2 d PP and at 20 ± 3 d PP. Blood plasma was analyzed for concentrations of albumin, haptoglobin, reactive oxygen and nitrogen species (RONS), and antioxidant potential (AOP), with lipopolysaccharide-binding protein (LBP) and TNFα evaluated during the TP only. Oxidative stress index (OSi) was calculated as RONS/AOP. Abundance of mRNA from genes involved in inflammation and glucose metabolism in liver and genes involved in lipogenesis in adipose tissue were determined. Data were analyzed separately for the TP and CO periods. During the TP, treatments interacted to affect concentrations of TNFα, haptoglobin, and LBP, with HMC increasing their concentrations for HS (9.38 vs. 7.45 pg/mL, 0.45 vs. 0.37 mg/mL, and 5.94 vs. 4.48 µg/mL, respectively) and decreasing their concentrations for LS (4.76 vs. 12.9 pg/mL, 0.27 vs. 0.41 mg/mL, and 4.30 vs. 5.87 µg/mL, respectively) compared with DGC. Effects of treatments diminished over time for LBP and haptoglobin with no differences by the end of the TP and no main CO effects of treatment for haptoglobin. The opposite treatment interaction was observed for albumin, with HMC tending to decrease its concentration for HS (3.24 vs. 3.34 g/dL) and increase its concentration for LS (3.35 vs. 3.29 g/dL) compared with DGC, with no carryover effect. Feeding DGC increased the OSi during the first week of the TP compared with HMC, with this effect diminishing over time; during the CO period HMC increased OSi for HS and decreased it for LS compared with DGC, with this effect diminishing toward the end of CO. Feeding HMC increased the abundance of genes associated with inflammation and gluconeogenesis in liver for HS and decreased it for LS compared with DGC. Feeding HS increased the mRNA abundance of genes associated with adipose tissue lipogenesis compared with LS. Results during the TP suggest that feeding LS-DGC and HS-HMC elicited a more pronounced inflammatory response and induced an upregulation of genes associated with inflammation and gluconeogenesis in liver, without effects on OSi, but effects on plasma markers of inflammation diminished during the CO period.

Efficient poultry manure management: anaerobic digestion with short hydraulic retention time to achieve high methane production.

The efficient treatment or appropriate final disposal of poultry manure (PM) to avoid serious environmental impacts is a great challenge. In this work, the optimization of a 2-stage anaerobic digestion system (ADS) for PM was studied with the aim of reaching a maximal methane yield with a short hydraulic retention time (HRT). Three activities were performed: The first activity, ADS 1, consisted of evaluating the effect of the substrate concentration and the HRT on the process, with a constant organic loading rate (OLR) of 3.66 ± 0.21 gVS L-1 d-1. The second activity, ADS 2, consisted of decreasing the HRT from 9.09 to 2.74 d with a constant substrate concentration. In the third activity, ADS 3, the substrate concentration was increased from 10.09 ± 1.41 to 35.25 ± 6.20 gVS L-1 with an average HRT of 4.66 ± 0.11 d. Maximal methane yields of 0.22, 0.21, and 0.22 LCH4 gVS-1 were reached for ADS 1, ADS 2, and ADS 3, respectively, at a low HRT (3.38 to 4.66 d) and high free ammonia concentration (between 323.05 ± 56.48 and 460.93 ± 135.40 mgN-NH3 L-1). These methane yields correspond to the production of 40.36 and 42.28 cubic meters of methane per ton of PM, respectively, and a laying hen produces between 47.45 and 54.75 kg of PM per year in Chile. Finally, this is the first study of the separate and combined effects of OLR, HRT and substrate concentration on the anaerobic digestion of PM. The results demonstrate the technical feasibility of the two-stage ADS treatment of PM with a short HRT; the system tolerates variations in the total ammonia nitrogen concentration of PM throughout the year and achieves a high methane yield when the correct operational conditions are selected.

Technological Advances in Wound Treatment of Exotic Pets.

Although most research about the use of technological advances for wound healing was performed in laboratory animals but oriented to human medicine, recent technological advances allowed its application not only to small animals but also to exotic pets. This article reviews the literature available about some of these techniques (negative wound pressure therapy, photobiomodulation [laser therapy], electrical stimulation therapy, therapeutic ultrasonography, hyperbaric oxygen therapy), and other advances in wound management (skin expanders, xenografts, and bioengineered autologous skin substitutes) in exotic pet species.

Evaluation of a single-flow continuous culture fermenter system for determination of ruminal fermentation and enteric methane production.

A 4-unit, single-flow continuous culture fermenter system was developed to assess in vitro nutrient digestibility, volatile fatty acid (VFA) concentration and daily enteric methane (CH4 ) production of ruminant diets. The objective was to develop a closed-vessel system that maintained protozoal populations and provided accurate predictions of total CH4 production. A diet of 50% orchardgrass (Dactylis glomerata L.) and 50% alfalfa (Medicago sativa L.) was fed during 4, 10-day periods (7-day adaptation and 3-day collection). Fermenters were fed 82 g of dry matter (DM)/day in four equal feedings. pH and temperature were taken every 2 min, and CH4 concentration was measured every 10 min. Samples for DM and protozoal counts were taken daily, and daily effluent samples were collected for determination of DM, VFA and NH3 -N concentrations. There was no effect (p > 0.17) of adaptation versus collection days on vessel and effluent DM, temperature or pH. Initial protozoal counts decreased (p < 0.01), but recovered to initial counts by the collection period. Total VFA, acetate, propionate and isobutyrate concentrations did not differ (p ≥ 0.13) among periods or days of the collection period. There was no difference (p ≥ 0.37) among days or periods in total daily CH4 production and CH4 production per g of OM, NDF, digestible OM or digestible NDF fed. Data collected throughout 4 experimental periods demonstrated that the system was able to reach a steady state in fermentation well within the 7-day adaptation period and even typically variable data (i.e., CH4 production) were stable within and across periods. While further research is needed to determine the relationship between this system and in vivo data, this continuous culture fermenter system provides a valid comparison of in vitro ruminal fermentation and enteric CH4 production of ruminant diets that can then be further validated with in vivo studies.

Effects of sample size on neutral detergent fiber digestibility of triticale forages using the Ankom DaisyII Incubator system.

Accurate and precise determinations of fiber digestibility are essential for proper diet formulation for dairy cows. Our objectives were 3-fold: (1) regress in vitro neutral detergent fiber digestibility (NDFD) values from 48 triticale forages determined at multiple endpoints ranging from 12 to 240 h with Ankom DaisyII Incubator system (Ankom Technology Corp., Macedon, NY) methods using 0.25- or 0.50-g sample sizes on concentrations of fiber-related analytes or growth stage; (2) directly compare NDFD values determined with 0.25- or 0.50-g sample sizes by Ankom methods after 12-, 24-, 30-, 48-, 144-, or 240-h incubations; and (3) compare NDFD values determined by Ankom methods after 30 and 48 h of incubation with those determined by traditional sealed-tube procedures obtained from a commercial laboratory. Generally, plant growth stage, which was quantified with a linear model suitable for serving as an independent regression variable, proved to be a better predictor variable for NDFD than neutral detergent fiber or acid detergent lignin. For direct comparisons of 0.25- and 0.50-g sample sizes using Ankom methods, the regression relationship for a 30-h incubation was explained by a linear model [Y = 1.206x - 1.1; coefficient of determination (R2) = 0.933], in which the slope differed from unity, but the intercept did not differ from 0. After a 48-h incubation, a linear model (Y = 1.014x + 7.1; R2 = 0.964) indicated that the slope did not differ from unity, but the intercept was >0. A linear regression (Y = 1.040x - 1.8; R2 = 0.861) of the 30-h incubation results obtained by Ankom methods using the 0.25-g sample size on those from the commercial laboratory indicated the slope and intercept did not differ from unity or 0, respectively. A similar relationship was obtained from the 48-h incubation (Y = 1.021x - 3.4; R2 = 0.866). Relationships were poorer when the 0.50-g sample size was used by Ankom methods, particularly for the 30-h incubation, where the slope (0.824) was less than unity. Generally, NDFD values were greater for the 0.25-g sample size by Ankom methods, especially with 24-, 30-, and 48-h incubation times, and agreement with traditional sealed-tube methods was improved with the smaller sample size. Synchronization of results between Ankom and traditional methods needs to be further verified across a wider range of forages and harvest/preservation methods before definitive recommendations can be made.

Treatment of duck house wastewater by a pilot-scale sequencing batch reactor system for sustainable duck production.

The objective of this study is trying to solve water pollution problems related to duck house wastewater by developing a novel duck house wastewater treatment technology. A pilot-scale sequencing batch reactor (SBR) system using different hydraulic retention times (HRTs) for treating duck house wastewater was developed and applied in this study. Experimental results showed that removal efficiency of chemical oxygen demand in untreated duck house wastewater was 98.4, 98.4, 87.8, and 72.5% for the different HRTs of 5, 3, 1, and 0.5 d, respectively. In addition, removal efficiency of biochemical oxygen demand in untreated duck house wastewater was 99.6, 99.3, 90.4, and 58.0%, respectively. The pilot-scale SBR system was effective and deemed capable to be applied to treat duck house wastewater. It is feasible to apply an automatic SBR system on site based on the previous case study of the farm-scale automatic SBR systems for piggery wastewater treatment.

Enteric methane production and ruminal fermentation of forage brassica diets fed in continuous culture.

The aim of the current study was to determine nutrient digestibility, VFA production, N metabolism, and CH4 production of canola (Brassica napus L.), rapeseed (B. napus L.), turnip (Brassica rapa L.), and annual ryegrass (Lolium multiflorum Lam.) fed with orchardgrass (Dactylis glomerata L.) in continuous culture. Diets were randomly assigned to fermentors in a 4 × 4 Latin square design using 7 d for adaptation and 3 d for collection. Diets were: 1) 50% orchardgrass + 50% annual ryegrass (ARG); 2) 50% orchardgrass + 50% canola (CAN); 3) 50% orchardgrass + 50% rapeseed (RAP); and 4) 50% orchardgrass + 50% turnip (TUR). Feedings (82 g DM/d) occurred four times daily throughout 4, 10-d periods at 730, 1030, 1400, and 1900 h. Methane samples were collected every 10 min using a photoacoustic gas analyzer (LumaSense Technologies, Inc.; Santa Clara, CA) during the last 3 d of the experiment. Effluent samples were collected on d 8, 9, and 10, composited by fermentor, and analyzed for VFA and pH as well as DM, OM, CP, and fiber fractions for determination of nutrient digestibility. Forage samples were analyzed for CP, NDF, ADF, minerals, and glucosinolate (GLS) concentrations. Data were analyzed using the GLIMMIX procedure of SAS. Apparent DM, OM, and NDF digestibilities and true DM and OM digestibilities were similar (P > 0.28) among diets (45.1, 63.2, 44.1, 67.1, and 87.2%, respectively). Total VFA (87.2 mol/100 mol), pH (6.47), and acetate (A: 44.6 mol/100 mol) were also not different (P > 0.20) among diets. The A:P (P = propionate) ratio was greater (P < 0.01) in ARG and CAN than RAP and TUR. Daily CH4 production was greater (P < 0.01) in ARG than all other diets (68.9 vs. 11.2 mg/d). Methane, whether expressed as g per g of OM, NDF, digestible OM, or digestible NDF fed was greatest (P < 0.01) in ARG but similar (P > 0.18) among brassica diets. A significant negative correlation was observed between total GLS and CH4 production. However, when multiple regression analysis on CH4 production was completed, neither total GLS nor individual GLS were a significant component of the model. Addition of brassicas provided similar nutrient digestibility to ARG while reducing daily CH4 production, potentially making brassicas an alternative for ARG in pasture-based ruminant diets.

Nutritional evaluation and ruminal fermentation patterns of kochia compared with alfalfa and orchardgrass hays and ephedra and cheatgrass compared with orchardgrass hay as alternative arid-land forages for beef cattle in two dual-flow continuous culture system experiments.

The objective was to evaluate the ruminal fermentation patterns of forage kochia (FK) compared with alfalfa hay (AH) and orchardgrass hay (OH) (Exp. 1), and ephedra (EPH) and immature cheatgrass (CG) compared with OH (Exp. 2), using a dual-flow continuous culture system. Two in vitro experiments were conducted, and in each experiment, treatments were randomly assigned to six dual-flow fermenters (1,223 ± 21 mL) in a replicated 3 × 3 Latin square design, with three consecutive periods of 10 d each, consisting of 7 d for diet adaptation and 3 d for sample collection. Each fermenter was fed a total of 72 g/d (DM basis) and treatments were as follows: Exp. 1: 1) 100% OH, 2) 100% AH, and 3) 100% dried FK. Exp. 2: 1) 100% OH, 2) 100% dried CG, and 3) 100% dried EPH. On day 8, 9, and 10, samples of solid and liquid effluent from each fermenter were taken for digestibility analysis, and subsamples were collected for NH3-N, VFA, and bacterial N determinations. Data were analyzed using the MIXED procedure of SAS. In Exp. 1, treatments did not affect DM, OM, and NDF digestibilities, total VFA and molar proportions of acetate, propionate, butyrate, and branched-chain VFA. True CP digestibility, ruminal NH3-N concentration, and total N, NH3-N, NAN, and dietary N flows (g/d) were greater (P < 0.05) for FK compared with the other forages. However, treatments did not affect bacterial efficiency. In Exp. 2, DM, OM, and CP digestibilities were greater (P = 0.01) for EPH, and NDF digestibility was greater (P < 0.05) for EPH and CG compared with OH. Ephedra had the highest (P < 0.05) pH and acetate:propionate ratio and the lowest (P < 0.05) total VFA concentration. Total VFA, ruminal NH3-N concentration, and NH3-N flow (g/d) were highest (P < 0.05) for CG. Total N flow and bacterial efficiency were highest (P < 0.05) for OH and CG, while the flows (g/d) of NAN, bacterial N, and dietary N were greater (P < 0.05) for OH compared with the other forages. Results indicate that when compared with AH and OH (Exp. 1), FK has similar ruminal fermentation patterns and may be an adequate alternative for beef cattle producers. Furthermore, when compared with OH (Exp. 2), immature CG may also be an adequate forage alternative. This is especially important for areas in which conventional forages may not grow well such as the U.S. arid-land. However, EPH should not be used as the sole forage due to its poor ruminal fermentation as evidenced by the lowest total VFA concentration and propionate molar proportion.

Effects of carbohydrate and nitrogen supplementation on fermentation of cheatgrass () in a dual-flow continuous culture system.

Cheatgrass (CG; ), an introduced winter annual grass, is an aggressive invader of the sagebrush community in the Western United States. Because of its greater flammability, mature CG constitutes a fire hazard leading to repeated wildfires. One fuel-reduction strategy is livestock grazing. The objective of this study was to evaluate the effects of urea, molasses, or a combination of urea and molasses supplementation of a CG-based diet on digestibility, microbial fermentation, bacterial protein synthesis, and nutrient flow using a dual-flow continuous culture system. Eight fermenters were used in a replicate 4 × 4 Latin square design with four 10-d experimental periods. Experimental treatments (DM basis) were 1) forage only (CON), 2) CG plus urea alone (URE; 1.36% urea), 3) CG plus molasses alone (MOL; 15.9% molasses), and 4) CG plus urea and molasses combined (URE+MOL; 1.28% urea plus 19.3% molasses). Each fermenter was fed 72 g/d of DM, and data were analyzed using the GLIMMIX procedure of SAS (SAS Inst. Inc., Cary, NC). The true digestibilities of NDF and ADF were not affected by diets ( > 0.05). Molasses-containing diets had greater true digestibility of OM ( = 0.02). However, true digestibility of CP was increased when molasses was fed alone ( < 0.01). Molasses-containing diets had lower pH ( < 0.01) and greater VFA concentrations ( < 0.01) compared to those of the other diets. The URE+MOL diet resulted in a greater VFA concentration ( < 0.01). Propionate concentration increased ( < 0.01), whereas acetate concentration decreased ( < 0.01) when molasses alone or in combination with urea was added to the diets. Supplying molasses alone resulted in greater ( = 0.03) total branched-chain VFA compared to the other diets. The concentration of NH-N and total N flow increased ( < 0.01) in response to urea supplementation and was greater ( < 0.01) when urea alone was supplemented in the diet. On the other hand, molasses-supplemented diets yielded more non-ammonia N ( < 0.01) and bacterial N ( = 0.04). Supplementation had no effect ( = 0.83) on bacterial efficiency. Results from this study indicate that the addition of urea and molasses in a CG-based diet could improve nutrient supply to animals, notably VFA supply and microbial N supply; however, in the levels tested in this study, it did not improve CG utilization as assessed by NDF digestion.

Putrescine: Essential factor for in vitro proliferation of Babesia bovis.

This study reports the effect of putrescine addition, either alone or in combination with insulin, transferrin and selenite (ITS), to serum-free Advanced DMEM/F12 (A-DMEM/F12) medium, on the in vitro culture of Babesia bovis and using a perfusion bioreactor to improve efficiency of the process. A B. bovis strain previously adapted to proliferate in serum-free medium (Bbovis-SF) was evaluated using eight increasing concentrations of putrescine. The percentage of parasitized erythrocytes (PPE) obtained from cultures supplemented with 0.101 mg/L was 6.23% compared with 2.3% for control cultures with M199 with Earle's salts (M199) and 40% serum. The combination of putrescine (0.101 mg/L) and a mixture of ITS (2000, 1100, and 1.34 mg/L, respectively) (Pu-ITS), in A-DMEM/F12 culture medium without serum yielded a maximum PPE of 17.26% compared to 2.58% in the control medium. This new formulation of culture medium, together with the use of a hollow-fiber perfusion bioreactor system (HFPBS), caused a substantial increase in the proliferation of B. bovis, yielding a maximum cumulative PPE of 118.8% after five days, compared to 58.6% in cultures treated with control medium M199 and 40% serum. We concluded that the addition of the ITS mixture and putrescine to the culture medium stimulated the proliferation of B. bovis in vitro. This new medium formulation, used in a HFPBS culture system, can be an effective, automated-prone system that can induce massive proliferation of B. bovis for use as a source of parasite antigens and immunogens.

An ovarian bioreactor for in vitro culture of the whole bovine ovary: a preliminary report.

BACKGROUND: Improved cancer therapeutics and enhanced cancer survivorship have emphasized the severe long-term side effects of chemotherapy. Specifically, studies have linked many chemotherapy agents with primary ovarian insufficiency, although an exact insult model has not yet been determined. To investigate and ultimately solve this problem, a novel device for extended study of mammalian ovaries in vitro was developed. METHODS: A bioreactor was fabricated for bovine ovarian culture that provides intravascular delivery of media to the ovary through isolation and cannulation of a main ovarian artery branch. Whole ovaries were cultured in vitro using three methods: (1) continuously supplied fresh culture media, (2) recirculated culture media, or (3) continuously supplied fresh culture media supplemented with 500 nM doxorubicin for 24 or 48 h. TUNEL assay was used to assess apoptotic cell percentages in the three groups as compared to uncultured baseline ovaries. RESULTS: The ovary culture method was shown to maintain cell viability by effectively delivering nutrient-enriched pH-balanced media at a constant flow rate. Lower apoptosis observed in ovaries cultured in continuously supplied fresh culture media illustrates that this culture device and method are the first to sustain whole bovine ovary viability for 48 h. Meanwhile, the increase in the percentage of cell apoptosis with doxorubicin treatment indicates that the device can provide an alternative model for testing chemotherapy and chemoprotection treatments to prevent primary ovarian insufficiency in cancer patients. CONCLUSIONS: An ovarian bioreactor with consistent culture media flow through an ovarian vasculature-assisted approach maintains short-term whole bovine ovary viability.

Effect of starchy or fibrous carbohydrate supplementation of orchardgrass on ruminal fermentation and methane output in continuous culture.

A 4-unit dual-flow continuous culture fermentor system was used to assess the effects of supplementing orchardgrass (Dactylis glomerata L.) with 2 levels [5 and 10% of total dry matter (DM) fed] of starchy (barley grain, BAR) or fibrous (beet pulp, BP) carbohydrate sources on nutrient digestibility, volatile fatty acid (VFA) production, bacterial protein synthesis, and CH4 output. Treatments were randomly assigned to fermentors in a 4×4 Latin square design with a 2×2 factorial arrangement using 7 d for microbial adaptation and 3 d for sample collection. Treatments included (1) 57g of DM orchardgrass + 3g of DM BAR, (2) 54g of DM orchardgrass + 6g of DM BAR, (3) 57g of DM orchardgrass + 3g of DM BP, or (4) 54g of DM orchardgrass + 6g of DM BP. Feedings occurred at 0900, 1030, 1400, and 1900h throughout four 10-d periods. Gas samples for CH4 analysis were collected 6 times daily at 0725, 0900, 1000, 1355, 1530, and 1630h. Fermentor samples for pH, NH3-N, and VFA analysis were taken on d 8, 9, and 10. Samples were also analyzed for DM, organic matter, crude protein, purines, neutral detergent fiber, and acid detergent fiber to determine nutrient digestibilities and estimation of bacterial protein synthesis. Apparent and true DM and organic matter digestibilities were not affected by supplement source. Apparent neutral and acid detergent fiber digestibilities were greater for BAR than BP. Conversely, apparent crude protein digestibility was greater for BP than BAR. Mean and maximum pH tended to be greatest for BAR than BP. Minimum pH was greater at the lower level (5% of diet DM) of supplementation. Barley produced greater concentrations of total VFA and acetate, whereas BP had greater daily outputs of CH4. Significant supplement type × level interactions were found for bacterial N flow and efficiency. Overall, supplementing orchardgrass with BP improved crude protein digestibility, reduced fiber digestibility and total VFA concentration, but increased CH4 output compared with BAR.

Tratamento de efluentes de abatedouro de frangos por meio de reator UASB seguido de filtro anaeróbio / Treatment of waste water poultry slaughter house through UASB reactor followed by anaerobic filter

O Brasil é um dos maiores produtores de aves do mundo e se destaca na exportação de carne de frango, que tem papel relevante na economia do país. Como resultado do crescimento da indústria de processamento do frango, há também um significativo aumento na geração de efluentes. Uma alternativa interessante para o tratamento desses resíduos são os reatores anaeróbios. Este trabalho teve como objetivo avaliar o desempenho do sistema reator UASB-Filtro anaeróbio no tratamento de efluentes de abate de frangos. O estudo foi desenvolvido em um abatedouro avícola, situado na região metropolitana de Belo Horizonte, Minas Gerais. O sistema de tratamento foi composto por peneira estática, caixa de gordura, reator UASB e filtro anaeróbio. A vazão média diária de efluentes do abatedouro foi de 30m³/dia. A carga orgânica aplicada no reator UASB foi de 0,71kg DQO/m³.d. O monitoramento do sistema foi realizado por meio de análises físico-químicas do afluente e dos efluentes da caixa de gordura, reator UASB e filtro anaeróbio. Os parâmetros avaliados foram: temperatura, pH, demanda química de oxigênio (DQO), demanda bioquímica de oxigênio (DBO), sólidos suspensos totais (SST), e sólidos suspensos voláteis (SSV). Os valores efluentes médios encontrados para DBO, DQO e SST foram 106, 290 e 56mg/L, respectivamente. A eficiência de remoção do sistema para DBO variou de 83 a 99%, e para DQO de 50 a 98%. A eficiência de remoção média de SST e SSV foi de 92%. Conclui-se que o sistema reator UASB seguido de filtro anaeróbio apresentou alto desempenho na remoção de sólidos e carga orgânica. Dessa forma, esses reatores tornam-se uma alternativa viável para o tratamento de águas residuárias de frigoríficos, já que oferecem bons resultados de remoção de poluentes.

Brazil is one of the largest poultry producers in the world and stands out in exportation of chicken meat, which plays an important role in the economy. As a result of the growth of the poultry processing industry, there is also a significant increase in the generation of wastewater. An interesting alternative for the treatment of such wastes is anaerobic. This study aimed to evaluate the performance of the system UASB reactor-anaerobic filter on wastewater treatment of slaughtering chickens. The study was conducted in a poultry slaughterhouse located in the metropolitan region of Belo Horizonte, Minas Gerais. The treatment system was composed of static sieve, grease trap, UASB reactor and anaerobic filter. The daily average flow of effluent from the abattoir was 30m³/day. The organic load applied to the UASB reactor was 0.71kg COD/m³.d. The monitoring system was performed by means of physico-chemical analysis of the influent and effluent of fat, UASB reactor and anaerobic filter. The parameters evaluated were: temperature, pH, chemical oxygen demand (COD), biochemical oxygen demand (BOD), total suspended solids (SS) and volatile suspended solids (VSS). The mean values for effluent BOD, COD and SS were 106, 290 and 56mg/L, respectively. The removal efficiency of BOD to the system ranged 83-99% for COD and 50-98%. The average removal efficiency of SS and SSV were 92%. We conclude that the system UASB reactor followed by anaerobic filter showed high performance in removing solids and organic load. Thus, these reactors become a viable alternative for treating wastewater slaughterhouses, offering good results in removing pollutants.

Effect of sprouted barley grain supplementation of an herbage-based or haylage-based diet on ruminal fermentation and methane output in continuous culture.

A 4-unit dual-flow continuous-culture fermentor system was used to assess the effect of supplementing 7-d sprouted barley (SB) or barley grain (BG) with an herbage-based or haylage-based diet on nutrient digestibility, volatile fatty acid (VFA) profiles, bacterial protein synthesis, and methane (CH4) output. Treatments were randomly assigned to fermentors in a 4 × 4 Latin square design with a 2 × 2 factorial arrangement using 7 d for diet adaptation and 3 d for sample collection. Experimental diets were (1) 55.5 g of herbage dry matter (DM) + 4.5 g of SB DM, (2) 56.0 g of herbage DM + 4.0 g of BG DM, (3) 55.5 g of haylage DM + 4.5 g of SB DM, and (4) 56.0 g of haylage DM + 4.0 g of BG DM. Forages were fed at 0730, 1030, 1400, and 1900 h, whereas SB and BG were fed at 0730 and 1400 h. Gas samples for CH4 analysis were collected at 0725, 0900, 1000, 1355, 1530, and 1630 h on d 8, 9, and 10. Fluid samples were taken once daily on d 8, 9, and 10 for pH measurements and for ammonia-N and VFA analysis and analyzed for DM, organic matter, crude protein, neutral detergent fiber, and acid detergent fiber for determination of nutrient digestibilities and estimation of bacterial protein synthesis. Orthogonal contrasts were used to compare the effect of forage source (haylage vs. herbage), supplement (BG vs. SB), and the forage × supplement interaction. Apparent and true DM and organic matter digestibilities as well as apparent crude protein digestibility were not affected by forage source. However, true DM digestibility was greatest for diets supplemented with SB. Apparent neutral and acid detergent fiber digestibilities of herbage-based diets were higher than haylage-based diets but fiber digestibility was not affected by supplement. Diets supplemented with SB had higher mean and minimum pH than BG; however, maximum pH was not affected by diet. Supplementation with BG produced a greater concentration of total VFA compared with diets supplemented with SB. Haylage-based diets produced greater CH4 output compared with herbage-based diets but supplementation did not affect CH4 output. Efficiency of bacterial protein synthesis was greater for herbage-based diets compared with haylage-based diets, with no effect of supplementation. Overall, supplementation with SB marginally increased true DM digestibility of herbage- and haylage-based diets but did not affect fiber and crude protein digestibilities, CH4 output, and bacterial efficiency, compared with BG.

Effect of bioaugmentation on anaerobic wastewater treatment in the dairy industry.

The creation of a biofilm as a specialized biosystem, its development, and activity were studied at 3 critical control points of biofilter operation: start-up, transition from batch to sequencing batch regimen, and set-up of stable sequencing batch process. Five variants of biosystems were investigated with an inoculum from specially adapted real activated sludge, enriched with various combinations of 3 microbial preparations. A stable and working biofilm was developed in the phase of stabilized sequencing batch process. The differences among biodegradation effectiveness of the 5 variants were insignificant during that phase and the effect of the preparations was low. The effectiveness of organic removal for the 5 bioaugmentation approaches reached 60% for protein, 70% for chemical oxygen demand, and 97% for lactose. Commercial inocula did not improve final reactor performance over an inoculum from a municipal wastewater treatment plant alone.

Comparison of in vitro digestibility estimates using the DaisyII incubator with in vivo digestibility estimates in horses.

The objective of this study was to determine if in vitro methodologies developed for the Ankom Daisy(II) incubator could produce accurate estimates of in vivo equine DM digestibility (DMD) and NDF digestibility (NDFD) when equine feces were used as the inoculum source. Four mature geldings were utilized in a 4 × 4 Latin square design experiment with a 2 × 2 factorial arrangement of dietary treatments (timothy hay, alfalfa hay, timothy hay plus oats, and alfalfa hay plus oats), in which the geldings were individually housed and fed. During each 5-d total fecal collection period, feces were collected and composited daily and used to calculate in vivo digestibility. Digestion of the 4 treatment diets was evaluated in vitro using the Daisy(II) incubator. Each incubation vessel of the Daisy(II) was assigned to 1 of the horses and contained 18 filter bags (6 containing the assigned treatment hay, 6 containing hay-oat mix, and 6 containing oats). Three incubation periods were evaluated: 30, 48, and 72 h. Although the 30- and 48-h in vitro estimates were consistently less than the in vivo estimates, they ranked diets in the same order as the in vivo method. For the alfalfa oat diet, timothy diet, and the timothy oat diet, the mean 72-h in vitro DMD and in vivo DMD were not different (P = 0.1444). However, for the alfalfa diet, the DMD estimate from 72-h in vitro incubation was less than the in vivo estimate (P < 0.010). For NDFD, the timothy diet was the only diet, in which the mean 72-h in vitro NDFD estimate was not different than the in vivo estimate. However, the in vitro method correctly ranked the alfalfa-based diets as having greater NDFD estimates than the timothy-based diets. Of the 3 incubation periods, the 72-h period provided digestibility estimates most similar to the in vivo data. Using the methodologies described in this research, the Daisy(II) incubator and equine feces can be used to estimate in vivo DMD of horse feeds.

Endothelial bioreactor ameliorates endotoxemia sepsis in swine.

INTRODUCTION: Severe sepsis involves a complex response including the activation of lots of cells, inflammatory mediators, and the hemostatic system. Central to this process is an alteration of endothelial cell function. Therefore, we investigated whether an endothelial bioreactor (EBR) would provide a new therapeutic approach to this clinical disorder. METHODS: EBR was constructed using a cartridge which contained with nonwoven polytetrafluoroethylene seeded with porcine iliac artery endothelial cell (PIEC). Pigs were intravenously administered with 0.25 mg/kg lipopolysaccharide and immediately placed in an extracorporeal circuit with EBR or sham controls. RESULTS: Compared with the sham group, EBR therapy resulted in significantly higher mean arterial blood pressure and significantly lower plasma von Willebrand factor, endothelin-1 and scores of lung injury. These alterations were associated with a significant survival advantage in the EBR group. CONCLUSIONS: Timely intervention with EC therapy in a tissue-engineered bioreactor may improve cardiovascular performance and alter the natural history of endotoxemia sepsis.