Nutrient ileal digestibility evaluation of dried mealworm (Tenebrio molitor) larvae compared to three animal protein by-products in growing pigs.

OBJECTIVE: This study was to investigate the nutrient ileal digestibility of dried mealworm (Tenebrio molitor) larvae and compare with those of three animal protein by-products in growing pigs. METHODS: A total of 12 crossbred ([Landrace×Yorkshire]×Duroc) growing pigs with average body weights of 24.12±0.68 kg were surgically equipped with simple T-cannulas after being deprived of feed for 24 h according to published surgical procedures. These pigs had a recovery period of two weeks. A total of 12 pigs were assigned to individual metabolic crates and allotted to one of four treatments with 3 replicates in a fully randomized design. Dietary treatments included the following: i) Fish meal, corn-vegetable by-product basal diet+9.95% fish meal; ii) Meat meal, corn-vegetable by-product basal diet+9.95% meat meal; iii) Poultry meal, corn-vegetable by-product basal diet+9.95% poultry meal; iv) Tenebrio molitor, corn-vegetable by-product basal diet+9.95% dried Tenebrio molitor larvae. RESULTS: Results showed that the apparent ileal digestibility (AID) of Lys was higher (p<0.05) in pigs fed Tenebrio molitor diet than that in pigs fed fish meal diet. Pigs fed Tenebrio molitor diet showed increased (p<0.05) AID of His and Arg compared to pigs fed Fish meal or Meat meal diet. The AID of Cys was increased (p<0.05) in pigs fed poultry meal and Tenebrio molitor diets compared to that in pigs fish meal diet. Pigs fed meat meal, poultry meal, and Tenebrio molitor diets showed higher (p<0.05) standardized ileal digestibility (SID) of total energy compared to pigs fed fish meal diet. The SID of Arg was higher (p<0.05) in pigs fed Tenebrio molitor diet than that in pigs fed fish meal or meat meal diet. Furthermore, pigs fed poultry meal or Tenebrio molitor diets showed increased (p<0.05) SID of Cys compared to pigs fed fish meal diet. CONCLUSION: In conclusion, providing pigs with diets that contained Tenebrio molitor larvae meal improved AID and SID of nutrients as well as essential and non-essential amino acids. The digestibility of dried mealworm larvae protein and its utilization in vivo are also good. Therefore, dried mealworm larvae protein can be used as protein source at 10% level in growing pigs.

Effects of Saccharomyces cerevisiae fermentation products on dairy calves: Performance and health.

The aim of the study was to evaluate the effects of Saccharomyces cerevisiae fermentation products (SCFP) on performance and health of calves during the first 63 d of age. Sixty Holstein calves (30 males and 30 females) at 2 d of age were blocked by sex and date of birth then randomly assigned within blocks to 1 of 3 treatments. A texturized calf starter was fed ad libitum containing 0 (control), 0.5, or 1% SCFP (Original XPC, Diamond V, Cedar Rapids, IA) of DM. In addition, the supplemented calves were fed 1 g/d SCFP (SmartCare, Diamond V) in milk until d 30. All calves were fed 4 L of colostrum within 1 h of birth and were subsequently fed milk twice daily until weaned at 56 d of age. Male calves were harvested on d 56. Performance and health of weaned female calves were monitored until 63 d of age to determine the effect of preweaning treatment of SCFP on weaning stress. Starter intake, fecal scores, and medical treatments were recorded daily. Body weight measures and blood samples were collected on d 2, 28, 56, and 63. Serum was analyzed for blood urea nitrogen, fatty acids, insulin-like growth factor-1, glucose, and total protein. Oxidative biomarkers and total antioxidant capacity were also evaluated in the serum. Body weight, DMI, blood parameters, and oxidative biomarkers did not differ among treatments. Supplementation of SCFP lowered fecal scores in the pre- and postweaning periods. Saccharomyces cerevisiae fermentation products can be used to reduce the diarrhea in calves grown under normal commercial conditions.

Effects of Supplemental Levels of Saccharomyces cerevisiae Fermentation Product on Lactation Performance in Dairy Cows under Heat Stress.

The objectives of this study were to evaluate the effects of different supplemental levels of Saccharomyces cerevisiae fermentation product (SCFP; Original XP; Diamond V) on lactation performance in Holstein dairy cows under heat stress. Eighty-one multiparous Holstein dairy cows were divided into 27 blocks of 3 cows each based on milk yield (23.6±0.20 kg/d), parity (2.88±0.91) and day in milk (204±46 d). The cows were randomly assigned within blocks to one of three treatments: 0 (control), 120, or 240 g/d of SCFP mixed with 240, 120, or 0 g of corn meal, respectively. The experiment was carried out during the summer season of 2014, starting from 14 July 2014 and lasting for 9 weeks with the first week as adaption period. During the experimental period, average daily temperature-humidity index (measured at 08:00, 14:00, and 20:00) was above 68, indicating that cows were exposed to heat stress throughout the study. Rectal temperatures tended to decrease linearly (p = 0.07) for cows supplemented with SCFP compared to the control cows at 14:30, but were not different at 06:30 (p>0.10). Dry matter intake was not affected by SCFP supplementation (p>0.10). Milk yield increased linearly (p<0.05) with increasing levels of SCFP. Feed efficiency (milk yield/dry matter intake) was highest (p<0.05) for cows fed 240 g/d SCFP. Cows supplemented with SCFP gained (p<0.01) body weight, while cows in the control lost body weight. Net energy balance also increased linearly (p<0.01) with increasing levels of SCFP. Concentrations of milk urea nitrogen (p<0.01) decreased linearly with increasing levels of SCFP, while no difference (p>0.10) was observed among the treatments in conversion of dietary crude protein to milk protein yield. In summary, supplementation of SCFP alleviated the negative effect of heat stress in lactating Holstein dairy cows and allowed cows to maintain higher milk production, feed efficiency and net energy balance. Effects of SCFP were dose-dependent and greater effects were observed from higher doses.

Inflammatory response of a prostate stromal cell line induced by Trichomonas vaginalis.

While Trichomonas vaginalis, a cause of sexually transmitted infection, is known as a surface-dwelling protozoa, trichomonads have been detected in prostatic tissue from benign prostatic hyperplasia and prostatitis by immunoperoxidase assay or PCR. However, the immune response of prostate stromal cells infected with T. vaginalis has not been investigated. Our objective was to investigate whether T. vaginalis could induce an inflammatory response in prostate stromal cells. Incubation of a human prostate stromal myofibroblast cells (WPMY-1) with live T. vaginalis T016 increased expression of the inflammatory chemokines CXCL8 and CCL2. In addition, TLR4, ROS, MAPK and NF-κB expression increased, while inhibitors of TLR4, ROS, MAPKs and NF-κB reduced CXCL8 and CCL2 production. Medium conditioned by incubation of WPMY-1 cells with T. vaginalis stimulated the migration of human neutrophils and monocytes (THP-1 cells). We conclude that T. vaginalis increases CXCL8 and CCL2 production by human prostate stromal cells by activating TLR4, ROS, MAPKs and NF-κB, and this in turn attracts neutrophils and monocytes and leads to an inflammatory response. This study is the first attempt to demonstrate an inflammatory reaction in prostate stromal cells caused by T. vaginalis.

VP2 capsid domain of the H-1 parvovirus determines susceptibility of human cancer cells to H-1 viral infection.

Although H-1 parvovirus is used as an antitumor agent, not much is known about the relationship between its specific tropism and oncolytic activity. We hypothesize that VP2, a major capsid protein of H-1 virus, determines H-1-specific tropism. To assess this, we constructed chimeric H-1 viruses expressing Kilham rat virus (KRV) capsid proteins, in their complete or partial forms. Chimeric H-1 viruses (CH1, CH2 and CH3) containing the whole KRV VP2 domain could not induce cytolysis in HeLa, A549 and Panc-1 cells. However, the other chimeric H-1 viruses (CH4 and CH5) expressing a partial KRV VP2 domain induced cytolysis. Additionally, the significant cytopathic effect caused by CH4 and CH5 infection in HeLa cells resulted from preferential viral amplification via DNA replication, RNA transcription and protein synthesis. Modeling of VP2 capsid protein showed that two variable regions (VRs) (VR0 and VR2) of H-1 VP2 protein protrude outward, because of the insertion of extra amino-acid residues, as compared with those of KRV VP2 protein. This might explain the precedence of H-1 VP2 protein over KRV in determining oncolytic activity in human cancer cells. Taking these results together, we propose that the VP2 protein of oncolytic H-1 parvovirus determines its specific tropism in human cancer cells.

Lateral spread response monitoring during microvascular decompression for hemifacial spasm. Comparison of two targets of partial neuromuscular blockade.

AIM: The aim of the present study was to determine (1) whether successful intraoperative electromyography monitoring for lateral spread response (LSR) is possible with partial neuromuscular blockade (NMB) in subjects undergoing microvascular decompression (MVD) for hemifacial spasm and (2) the adequate level of NMB to achieve that goal. MATERIAL AND METHODS: A total of 61 patients in whom LSR was monitored during MVD were enrolled in the study. Patients were randomly allocated to two groups: group TOF in which the NMB target was maintenance of two train-of-four (TOF) counts and group T1 in which the NMB target was maintenance of a T1/Tc ratio of 50 % (T1: first twitch height of TOF and Tc: control twitch height). The adductor pollicis brevis muscle was used to monitor TOF responses. The frequency of successful LSR monitoring, defined as successful baseline establishment and maintenance of LSR until surgical decompression, was compared between the two groups. RESULTS: Of the 61 patients 2 were excluded from the study so that 30 patients in group TOF and 29 patients in group T1 were analyzed. The success rate of LSR monitoring was clinically acceptable and significantly higher in group T1 than in group TOF, i.e. n = 15 (50.0 %) in group TOF versus n = 24 (82.8 %) in group T1 (P = 0.008), corresponding to a 32.8 % higher success rate in group T1 than group TOF (95 % CI: 13.9-51.7 %). Mean vecuronium infusion dose was smaller and mean TOF count was higher in group T1 than group TOF with a TOF count = 2 (1) in group TOF versus 3 (1) in group T1 (P = 0.003). Mean sevoflurane and remifentanil infusion doses were not different between groups. There was no incidence of spontaneous movement during microscopy in either group. CONCLUSION: Maintenance of partial NMB with a target T1/Tc ratio of 50 % resulted in a clinically acceptable success rate of LSR monitoring and surgical condition during MVD. Maintenance of partial NMB with a target T1/Tc ratio of 50 % rather than TOF count of two during LSR monitoring for MVD can therefore be recommended.

Risk-based long-term screening for hepatocellular carcinoma recurrence after living donor liver transplantation.

BACKGROUND: This study sought to establish an actual risk-based long-term screening protocol for hepatocellular carcinoma (HCC) recurrence after liver transplantation (OLT). METHODS: The study was a retrospective review of medical records from 334 HCC patients who underwent primary living donor OLT and followed up for at least 5 years. RESULTS: Overall 10-year patient survival rate was 67.5%, with a 4.8% perioperative mortality. HCC recurred in 68/318 (21.4%) surviving patients over a mean follow-up of 77 months. HCC recurrence was 20.7% at 5 and 22.2% at 10 years. Annual recurrence rates were 11.4%, 6.6%, and 2.0% during the first, second, and third years, respectively. Among patients within Milan criteria, the annual incidence of HCC recurrence was highest during the first 3 years; thereafter only 6 sporadic recurrences were observed during next 8 years. Among subjects beyond Milan criteria, recurrence was common during, but not after 3 years. In 43 patients (63.2%) increased alpha-fetoprotein (AFP) was an initial indication to perform further imaging studies to diagnosis recurrence, whereas they were detected incidentally on protocol screening imaging among another 25 patients (36.8%) in the absence of an AFP rise. There was a close correlation between pretransplant AFP level and AFP increase after HCC recurrence. CONCLUSIONS: Patients beyond the Milan criteria require frequent tumor marker tests and imaging studies over the first 3 years; and those within Milan criteria require 10-years to follow-up primarily with tumor marker tests.

Enteric methane emission, diet digestibility, and nitrogen excretion from beef heifers fed sainfoin or alfalfa.

Effects of plant-bound condensed tannin (CT)-containing sainfoin vs. CT-free alfalfa (or low-CT alfalfa-sainfoin mixture), plant stage of maturity, and their interaction on enteric methane (CH4) emissions, diet digestibility, and N excretion were studied, using 8 ruminally cannulated beef heifers in 2 sequential short-term experiments (Exp. 1 and 2). In Exp. 1, first growth legumes were harvested daily and offered fresh to heifers. Heifers were assigned to 100% sainfoin or 80% alfalfa:20% sainfoin (as-fed basis). Responses were measured at early (late vegetative to early bud; stage 2 to 3) and late (early flower; stage 5) stage of maturity. In Exp. 2, the same legumes were harvested from second growth (late bud; stage 4) and offered to heifers as hay; 100% sainfoin or 100% alfalfa. In both experiments, heifers were fed once daily at 1× maintenance. When fed as fresh forage (Exp. 1), sainfoin, compared with the alfalfa-sainfoin blend, had greater digestibility of OM (74.7 vs. 70.9%; P = 0.02), yet tended to have lower CP digestibility (73.2 vs. 77.1%; P = 0.059). There was no difference between fresh legumes for CH4 emissions [25.9 g/kg DMI ± 4.02 SE; 8.5% of gross energy intake (GEI) ± 1.26 SE; or 36.8 g/kg digested OM ± 1.75 SE]. The fresh legumes were more digestible at early, rather than at late, maturity and, consequently, enteric CH4 (27.4 vs. 24.4 g/kg DMI; P < 0.004; 8.9 vs. 8.1% GEI; P < 0.008) was greater at early, rather than at later, growth. When fed as hay (Exp. 2), sainfoin, compared with alfalfa, had greater digestibility of OM (60.5 vs. 50.3%; P = 0.007), lower digestibility of CP (64.2 vs. 68.8%; P = 0.004), yet there was no difference between the legume hays for CH4 emissions (22.4 g/kg DMI ± 1.29 SD and 7.1% GEI ± 0.40 SD). However, on the basis of OM digested, CH4 emissions were lower for sainfoin than alfalfa hay (44.3 vs. 59.0 g/kg; P = 0.008). Percentage of total N excretion in urine was less for sainfoin compared with alfalfa, both for fresh legumes in Exp. 1 (74 vs. 78%; P = 0.017) or hay in Exp. 2 (64 vs. 72%; P < 0.001), and increasing maturity lowered urinary N excretion. In conclusion, feeding CT-containing sainfoin partially shifted N excretion from urine to feces, but it had little impact on enteric CH4 emissions from beef cattle fed at maintenance as compared with feeding either 80% alfalfa:20% sainfoin (fresh forages) or 100% alfalfa (hay). Feeding fresh legumes harvested between the late vegetative to early bud stage, compared with harvested at the early flower stage, increased N excreted in urine as well as enteric CH4 emissions from beef cattle fed at maintenance.

Synergistic effects of A1896, T1653 and T1762/A1764 mutations in genotype c2 hepatitis B virus on development of hepatocellular carcinoma.

The effects of genomic changes in hepatitis B virus (HBV) on the occurrence of hepatocellular carcinoma (HCC) are still unclear, especially in relation to the genotype of HBV. In this study, we examined the effects of genomic changes in HBV of genotype C2 on the development of HCC. A total of 318 patients with HBV-associated HCC and 234 patients with chronic hepatitis B (CHB) were studied. All of HCC cases were diagnosed histologically and treated with surgical resection. The whole of the X, S, basal core promoter (BCP) and precore regions of the viral genome from sera or liver tissues were sequenced. All subjects had HBV of genotype C2. The prevalence of the T1653 mutation in the X region and the A1896 mutation in the precore region of HBV was significantly higher in the HCC group than in the control CHB group (22% vs 11%, P = 0.003; 50% vs 23%, P < 0.001, respectively). Moreover, the T1762/A1764 mutations in the BCP region in combination with either T1653 or A1896 were more common in the HCC compared with the CHB group (BCP+X1653: 18% vs 11%, P = 0.05; BCP+PC, 40% vs 15%, P < 0.001, respectively). In multivariate analysis, T1653 and A1896 were revealed to be independent risk factors for HCC development. G1896A in the precore region and C1653T mutation in the X region of genotype C2 HBV are important risk factors for HCC development. Also, the A1762T/G1764A double mutation may act in synergy with C1653T to increase the risk of HCC in patients chronically infected with HBV genotype C2.

Cancer upregulated gene 2, a novel oncogene, confers resistance to oncolytic vesicular stomatitis virus through STAT1-OASL2 signaling.

We have recently found a novel oncogene, named cancer upregulated gene 2 (CUG2), which activates Ras and mitogen-activated protein kinases (MAPKs), including ERK, JNK and p38 MAPK. Because activation of these signaling pathways has previously been shown to enhance cancer cell susceptibility to oncolysis by certain viruses, we examined whether vesicular stomatitis virus (VSV) could function as a potential therapeutic agent by efficiently inducing cytolysis in cells transformed by CUG2. Unexpectedly, NIH3T3 cells stably expressing CUG2 (NIH-CUG2) were resistant to VSV because of the activation of signal transducers and activators of transcription 1 (STAT1). The result was supported by evidence showing that suppression of STAT1 with short interference RNA (siRNA) renders cells susceptible to VSV. Furthermore, 2'-5' oligoadenylate synthetase-like (OASL) 2 was the most affected by STAT1 expression level among anti-viral proteins and furthermore suppression of OASL2 mRNA level caused NIH-CUG2 cells to succumb to VSV as seen in NIH-CUG2 cells treated with STAT1 siRNA. In addition, Colon26L5 carcinoma cells stably expressing CUG2 (Colon26L5-CUG2) exhibited resistance to VSV, whereas Colon26L5 stably expressing a control vector yielded to VSV infection. Moreover, Colon26L5-CUG2 cells stably suppressing STAT1 succumbed to VSV infection, resulting in apoptosis. Taken together, we propose that VSV treatment combined with the selective regulation of genes such as STAT1 and OASL2 will improve therapeutic outcomes for CUG2-overexpressing tumors.

[Recovery from parkinsonism with N-acetylcysteine-differentiated neurons].

The upregulation of dopaminergic neuronal differentiation is necessary for stem cell therapy in Parkinson's disease (PD). In this study, neuronal differentiation efficiency increased by more than 2 times in P19 embryonic stem cells (ESCs) induced by N-acetylcysteine (NAC) and retinoic acid (RA) as compared to RA alone, with suppressed glial differentiation. The majority of NAC-treated stem cells grafted into brains of PD mice differentiated into dopaminergic neurons and persisted well for 6 weeks. Parkinsonism was also greatly improved after grafting NAC-treated cells in comparison to cells treated with only RA. Our results strongly suggest that NAC treatment may be an effective strategy for generating stem cells fated to become dopaminergic neurons for PD clinical therapy.

N-acetylcysteine enhances neuronal differentiation of P19 embryonic stem cells via Akt and N-cadherin activation.

We examined whether N-acetylcysteine (NAC) enhanced embryonic body (EB) formation and neuronal differentiation in terms of EB formation, neuronal marker (microtubule-associated protein 2; MAP-2) expression, and neuron maturation using P19 embryonic stem cells. The size and numbers of EBs were greatly increased, together with the up-regulated N-cadherin expression. Also, MAP-2 expression and neurite outgrowth were much increased with activation of serine/threonine protein kinase (Akt) and blocked by addition of an Akt inhibitor (LY294002). Our results suggested that NAC increased EB formation by up-regulating the N-cadherin expression. Furthermore, NAC-enhanced neuronal differentiation was mediated by activation of Akt.

Effects of partially replacing dietary starch with dry glycerol in a lactating cow diet on ruminal fermentation during continuous culture.

The effects of dry glycerol as a partial replacement for dietary starch in a lactating cow diet on ruminal fermentation and bacterial protein synthesis were evaluated using 4 single-flow, continuous-culture fermentors (ranging from 1,015 to 1,040 mL in volume). The basal lactating cow diet was formulated to have partial contents of dietary starch provided from a corn starch supplement [at 12.37% diet dry matter (DM)], which was partially or completely replaced by a dry glycerol product. Both the corn starch supplement and dry glycerol product contained 65% of pure corn starch or glycerol, respectively. The final inclusion rate for pure glycerol was at 0, 3, 5, or 8% of DM in the basal diet. The experiment was conducted using a 4 × 4 Latin square design with four 9-d periods, with the first 6 d for adaptation and last 3 d for sampling. Fermentors were inoculated with 1L of ruminal fluid and 25 g of ruminal digesta from a ruminally cannulated cow receiving a lactation total mixed ration (16% crude protein, 32% neutral detergent fiber, and 25% starch; DM basis). Each fermentor was fed 75 g of DM of its respective experimental diet daily in 3 equal portions (at 0800, 1400, and 2000 h). Liquid dilution rate of the fermentors was maintained at 10%/h and solids retention time was set at 24 h. Fermentation fluid and the effluent from each fermentor were sampled once daily (at 1330 h) from d 7 to 9 of each period and pooled by period. Postprandial ruminal fermentation was studied by sampling the fermentors hourly for 5 h after the 0800 h feeding on d 9 of each period. The total fermentation contents were harvested at the end of the period for estimations of bacterial protein synthesis. Replacing corn starch with dry glycerol linearly increased the proportions of propionate and valerate at the expense of acetate in the fermentation fluid measured daily or for the first 5h after feeding. Replacing corn starch with dry glycerol also linearly increased the digestibility of dietary neutral detergent fiber without a change on the flow or efficiency of bacterial protein synthesis during continuous culture. Results indicate that glycerol as a dry product can replace dietary starch as corn starch at a level of up to 8% of DM in the diet without negatively affecting ruminal fermentation and digestibility during continuous culture.

Effects of magnesium sulphate on postoperative coagulation, measured by rotational thromboelastometry (ROTEM(®)).

We investigated the effects of magnesium sulphate on blood coagulation profiles using rotational thromboelastometry in gynaecological patients undergoing pelviscopic surgery. Patients were randomly allocated to the magnesium group (n = 20) or control group (n = 20). The magnesium group received magnesium sulphate (50 mg.kg(-1) followed by continuous infusion of 15 mg.kg(-1).h(-1)), whereas the control group received the same volume of isotonic saline according to the same methods. Mean (SD) postoperative serum magnesium levels were 1.58 (0.17) mmol.l(-1) in the magnesium group compared with 0.98 (0.06) mmol.l(-1) in the control group (p < 0.001). Postoperative clotting time, clot formation time, α-angle and maximum clot firmness of INTEM, and clot formation time, α-angle, and maximum clot firmness of EXTEM were significantly different between the two groups (p < 0.05). Intra-operative infusion of magnesium sulphate seems to attenuate postoperative hypercoagulability by maintaining magnesium levels at the upper limit of the normal range.

A fibrolytic enzyme additive for lactating Holstein cow diets: ruminal fermentation, rumen microbial populations, and enteric methane emissions.

The objective was to determine if supplementing a dairy cow diet with an exogenous fibrolytic enzyme additive (Econase RDE; AB Vista, Marlborough, Wiltshire, UK) altered fermentation, pH, and microbial populations in the rumen or enteric methane (CH(4)) emissions. In a companion study, this enzyme additive improved efficiency of fat-corrected milk production in a dose-dependent manner by up to 11% for early lactation dairy cows. Nine ruminally cannulated, lactating Holstein cows were used in a replicated 3 × 3 Latin square design with 21-d periods. Dietary treatments were 0 (control), 0.5 (low), and 1.0 (high) mL of enzyme/kg of total mixed ration dry matter. Rumen contents were collected on 2 d (d 15 and 19), ruminal pH was measured continuously for 6 d (d 13 to 18) by using an indwelling system, and enteric CH(4) production was measured for 3 d (d 16 to 18) using the sulfur hexafluoride tracer gas technique. The enzyme additive did not alter volatile fatty acids, NH(3), pH, or population densities of total protozoa, bacteria, and methanogens in ruminal fluid. However, population densities of certain bacteria, calculated as copy number of species-specific 16S-rRNA, were affected by enzyme treatment. Population density of Ruminobacter amylophilus was increased and that of Fibrobacter succinogenes tended to be increased by the high enzyme treatment. Selenomonas ruminantium tended to increase linearly with increasing levels of enzyme in the diet, although its population density was only numerically increased by the high enzyme treatment. Streptococcus bovis, however, tended to be decreased by the low enzyme treatment. Increasing the level of enzyme supplement in the diet also linearly increased enteric CH(4) production, even when adjusted for feed intake or milk production (19.3, 20.8, and 21.7 g of CH(4)/kg of dry matter intake or 12.9, 13.6, and 15.1g of CH(4)/kg of milk for the control, low, and high enzyme treatments, respectively). This shift in ruminal bacterial communities and higher CH(4) emissions could imply increased ruminal digestion of feed, which needs to be substantiated in longer term studies.

Relationship between rumen methanogens and methane production in dairy cows fed diets supplemented with a feed enzyme additive.

AIMS: To investigate the relationship between ruminal methanogen community and host enteric methane (CH(4) ) production in lactating dairy cows fed diets supplemented with an exogenous fibrolytic enzyme additive. METHODS AND RESULTS: Ecology of ruminal methanogens from dairy cows fed with or without exogenous fibrolytic enzymes was examined using PCR-denaturing gradient gel electrophoresis (PCR-DGGE) analyses and quantitative real-time PCR (qRT-PCR). The density of methanogens was not affected by the enzyme additive or sampling times, and no relationship was observed between the total methanogen population and CH(4) yield (as g per head per day or g kg(-1) DMI). The PCR-DGGE profiles consisted of 26 distinctive bands, with two bands similar to Methanogenic archaeon CH1270 negatively correlated, and one band similar to Methanobrevibacter gottschalkii strain HO positively correlated, with CH(4) yield. Three bands similar to Methanogenic archaeon CH1270 or Methanobrevibacter smithii ATCC 35061 appeared after enzyme was added. CONCLUSIONS: Supplementing a dairy cow diet with an exogenous fibrolytic enzyme additive increased CH(4) yield and altered the composition of the rumen methanogen community, but not the overall density of methanogens. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first study to identify the correlation between methanogen ecology and host CH(4) yield from lactating dairy cows.

Differing effects of 2 active dried yeast (Saccharomyces cerevisiae) strains on ruminal acidosis and methane production in nonlactating dairy cows.

Fifteen ruminally cannulated, nonlactating Holstein cows were used to measure the effects of 2 strains of Saccharomyces cerevisiae, fed as active dried yeasts, on ruminal pH and fermentation and enteric methane (CH(4)) emissions. Nonlactating cows were blocked by total duration (h) that their ruminal pH was below 5.8 during a 6-d pre-experimental period. Within each block, cows were randomly assigned to control (no yeast), yeast strain 1 (Levucell SC), or yeast strain 2 (a novel strain selected for enhanced in vitro fiber degradation), with both strains (Lallemand Animal Nutrition, Montréal, QC, Canada) providing 1 × 10(10) cfu/head per day. Cows were fed once daily a total mixed ration consisting of a 50:50 forage to concentrate ratio (dry matter basis). The yeast strains were dosed via the rumen cannula daily at the time of feeding. During the 35-d experiment, ruminal pH was measured continuously for 7 d (d 22 to 28) by using an indwelling system, and CH(4) gas was measured for 4 d (d 32 to 35) using the sulfur hexafluoride tracer gas technique (with halters and yokes). Rumen contents were sampled on 2 d (d 22 and 26) at 0, 3, and 6h after feeding. Dry matter intake, body weight, and apparent total-tract digestibility of nutrients were not affected by yeast feeding. Strain 2 decreased the average daily minimum (5.35 vs. 5.65 or 5.66), mean (5.98 vs. 6.24 or 6.34), and maximum ruminal pH (6.71 vs. 6.86 or 6.86), and prolonged the time that ruminal pH was below 5.8 (7.5 vs. 3.3 or 1.0 h/d) compared with the control or strain 1, respectively. The molar percentage of acetate was lower and that of propionate was greater in the ruminal fluid of cows receiving strain 2 compared with cows receiving no yeast or strain 1. Enteric CH(4) production adjusted for intake of dry matter or gross energy, however, did not differ between either yeast strain compared with the control but it tended to be reduced by 10% when strain 2 was compared with strain 1. The study shows that different strains of S. cerevisiae fed as active dried yeasts vary in their ability to modify the rumen fermentative pattern in nonlactating dairy cows. Because strain 2 tended (when compared with strain 1) to lower CH(4) emissions but increase the risk of acidosis, it may be prudent to further evaluate this strain in cattle fed high-forage diets, for which the risk of acidosis is low but CH(4) emissions are high.

Inclusion of flaxseed in hay- and barley silage diets increases alpha-linolenic acid in cow plasma independent of forage type.

Feeding flaxseed to cattle may be a means of increasing omega-3 fatty acid levels in ruminant products, but possible interactions with conserved forages have not been investigated. Twelve Holstein cows were used in a replicated 4 × 4 Latin Square experiment. Cows were fed one of four 50:50 forage:concentrate diets (DM basis): hay (hay control, HC), hay plus 15% ground flaxseed (hay-flaxseed, HF), barley silage (silage control, SC), and barley silage plus 15% ground flaxseed (silage-flaxseed, SF). Plasma concentrations of alpha-linolenic acid (ALA) did not differ between SC and HC diets. Flaxseed increased ALA (P < 0.05), but levels were not influenced by forage type. Flaxseed slightly increased 18:2n-6 (P < 0.05) and some n-6 and n-3 elongation and desaturation products, particularly arachidonic acid (ARA) and eicosapentaenoic acid (EPA). Flaxseed also increased C18:0 (P < 0.05) with this increase being greater (P < 0.01) for cows fed SF than HF. Feeding flaxseed also increased plasma C18:1-trans isomers (P < 0.01), predominantly vaccenic acid (VAA, 18:1-t11), with this increase being greater (P < 0.05) in cows fed HF than SF. Although conjugated linoleic acid (CLA) was increased (P < 0.001) with flaxseed it was not influenced by forage type (P = 0.06). Overall, feeding flaxseed increased plasma ALA, EPA, ARA and CLA independently of forage type. Feeding flaxseed with silage, however, resulted in more 18:0, while feeding flaxseed with hay resulted in greater accumulations of plasma 18:1-trans isomers mainly in the form of VAA.