Detection of free-living amoebae in domestic cats with and without naturally-acquired keratitis.

Pathogenic free-living amoebae, most notably Acanthamoeba spp., are important pathogens of the human cornea. The importance of infection with free-living amoebae in cats with keratitis is currently unclear. The aim of this study was to determine the frequency of amoeba detection in corneas of cats with naturally-acquired keratitis and in the ocular surface microflora of cats without ocular disease. Clinical ophthalmic and in vivo corneal confocal microscopic examinations were performed on 60 cats with keratitis. Corneal scrapings were analyzed by amoeba culture; cytological evaluation; and Acanthamoeba, Hartmannella, and Vahlkampfia PCR assays. Following ophthalmic examination, conjunctival specimens collected from 60 cats without clinically apparent ocular disease were analyzed similarly. In one cat with ulcerative keratitis, amoeba cysts and trophozoites were detected by in vivo corneal confocal microscopy; an Acanthamoeba sp. was isolated from corneal specimens and detected by Acanthamoeba PCR assay; and suppurative corneal inflammation was present cytologically. An Acanthamoeba sp. was isolated from conjunctival specimens from one cat without clinically apparent ocular disease, but with suppurative inflammation demonstrated cytologically. Both Acanthamoeba isolates belonged to the T4 genotype. Naegleria-like amoebae were isolated in samples from two cats with keratitis and seven cats without clinical ocular disease, but amoebae were not detected by the other assays in these samples. Amoeba detection by culture was significantly (P = 0.01) associated with cytologically diagnosed corneoconjunctival inflammation. This study identified naturally-acquired Acanthamoeba keratitis in cats. Detection of Naegleria-like amoebae in samples from cats with and without keratitis is of uncertain pathological significance.

Effects of nitrate adaptation by rumen inocula donors and substrate fiber proportion on in vitro nitrate disappearance, methanogenesis, and rumen fermentation acid.

A study was conducted to evaluate the main effects of dietary nitrate adaptation by cattle and alfalfa cell wall to starch ratio in in vitro substrates on nitrate disappearance and nitrite and volatile fatty acid (VFA) concentrations, as well as hydrogen (H2) and methane (CH4) accumulations. Rumen fluid from steers fed diets containing urea or nitrate was added into in vitro incubations containing sodium nitrate as the sole nitrogen source and 20 cell wall : 80 starch or 80 cell wall : 20 starch as the carbohydrate source. The results showed that during 24 h incubation, rumen fluid inoculums from steers adapted to dietary nitrate resulted in more rapid nitrate disappearance by 6 h of incubation (P < 0.01), no significant effect on nitrite concentration and diminished CH4 accumulation (P < 0.05). Cell wall to starch ratio did not affect nitrate disappearance, CH4 accumulation and total VFA concentration. The higher cell wall ratio had the lower total gas production and H2 concentration (P < 0.05). Ammonia-N (NH3-N) concentration increased because of adaptation of donors to nitrate feeding (P < 0.05). Nitrate adaptation did not alter total VFA concentration, but increased acetate, and decreased propionate and butyrate molar proportions (P < 0.01).

Effect of water and feed withdrawal and health status on blood and serum components, body weight loss, and meat and carcass characteristics of Holstein slaughter cows.

During marketing, cattle may be exposed to periods of water deprivation. The impact of water and feed access and health status on the physiological well-being and carcass characteristics of Holstein slaughter cows during preslaughter marketing was studied through analysis of serum components, BW loss percentage, and fresh meat composition. Ninety-one multiparous Holstein cows (609 ± 89 kg mean BW, 2.9 ± 0.5 mean BCS, varying stage of lactation) were purchased over 3 wk in 3 groups (n = 31, 29, and 31) at a terminal market in central Wisconsin. Each cow was screened to determine health status (sick or not sick) and randomly assigned to 1 of 3 water and feed withdrawal treatment pens (AL, ad libitum access to water for 36 h; 18H, 18 h of ad libitum access to water followed by 18 h of water withdrawal; 36H, 36 h of water withdrawal; all 3 treatments included 36 h of feed withdrawal) in a randomized complete block arrangement with repeated measures for serum components. Blood samples were collected by tail venipuncture at 0, 9, 18, 27, and 36 h of each treatment. Ambient temperatures were 1.9 ± 6.2°C during the trial period, which occurred over a 3-wk period in March and April 2007 near Arlington, WI. No difference (P > 0.05) was observed in mean serum cortisol in AL (18.41 ± 2.17 ng/mL) or 36H (22.98 ± 2.17 ng/mL). Mean serum glucose was greater (P < 0.05) in 36H pens (78.15 ± 0.77 mg/dL) than AL (75.91 ± 0.77 mg/dL). Mean serum creatinine was greater (P < 0.05) in 36H pens (0.71 ± 0.03 mg/dL) than AL (0.60 ± 0.03 mg/dL). The 36H pens also displayed increased (P < 0.05) serum albumin, anion gap, Ca, Cl, Na, cholesterol, and aspartate aminotransferase over AL. Greater (P < 0.05) mean percentage BW loss was observed in 36H pens (5.2 ± 0.6%) than AL (3.1 ± 0.6%). Mean muscle protein (%) was greater (P < 0.05) in 36H (22.2 ± 0.4%) than 18H (21.3 ± 0.4%). Mean muscle moisture (%) was greater (P < 0.05) in AL and 18H (75.3 ± 0.4% and 75.2 ± 0.4%) than 36H. Mean 24-h pH values were 5.92 (AL), 5.92 (18H), and 5.81 (36H; SE = 0.04) and were not different (P < 0.05). Observed pH and color values indicated a borderline dark-cutter state across all cattle in the study, regardless of water and feed access treatment. Based on these results, water and feed withdrawal in lairage should not exceed 18 h during the marketing of Holstein slaughter cows acclimated to springtime conditions to maintain BW, serum component concentrations, and fresh meat composition.

Finishing steers with diets based on corn, high-tannin sorghum, or a mix of both: feedlot performance, carcass characteristics, and beef sensory attributes.

The objective of this work was to compare feedlot performance, carcass characteristics, and beef sensory attributes from steers finished with diets based on corn, high-tannin sorghum (HTS), and a mix of both grains. Angus crossbred steers (n = 11 steers per treatment, initial BW = 404 +/- 18 kg) were finished on diets containing 765 g/kg of DM of corn, HTS, or a 1:1 mix of corn and HTS. Final BW (P < 0.01), ADG (P < 0.001), and G:F (P < 0.01) were reduced in steers fed HTS when compared with steers fed corn. Steers fed the mixed diet had greater G:F than the average between corn and HTS diets (P = 0.04), which indicated that mixing corn and HTS had positive associative effects. Estimated NE(m) of HTS was 1.91, and estimated NE(g) was 1.35 Mcal/kg of DM. Hot carcass weight (P < 0.01), trimmed carcass weight (P < 0.01), yield grade (P = 0.04), and 12th-rib fat thickness (P = 0.01) were less in steers fed HTS than in those fed corn. Estimated percentage of HCW as boneless, closely trimmed retail cuts was greater in steers fed HTS compared with those fed corn (P = 0.02) but, due to the decreased HCW, estimated amount of boneless, closely trimmed retail cuts was less in steers fed HTS than in those fed corn (P = 0.03). Steers fed HTS had greater pH on LM (P = 0.02) than steers fed corn, but the difference was small (5.42 +/- 0.02 vs. 5.36 +/- 0.02, respectively) and within the range of normal beef pH. Diet had no effect on Warner-Bratzler shear values (P > or = 0.72). Multivariate ANOVA indicated a difference in sensory attributes of beef from corn and HTS steers (Wilks' Lambda, P = 0.04). When evaluating each sensory attribute independently, panelists found beef from steers fed HTS to be less juicy (P < 0.01), less tender (P = 0.03), and more cooked (P < 0.01) than beef from animals fed corn. Data from this study indicated that by using a 1:1 mix of HTS and corn it is possible to finish steers to similar BW and carcass quality as by using a corn-based diet. Also, total replacement of corn by HTS in the diets produced lighter and leaner animals. Total replacement of corn by HTS in the finishing diet changed beef sensory attributes, reducing the perception of beef tenderness and juiciness by panelists but without changing instrumental tenderness.

Finishing steers with diets based on corn, high-tannin sorghum or a mix of both: Color and lipid oxidation in beef.

We tested the hypothesis that feeding high-tannin sorghum (HTS) to steers would produce beef more resistant to oxidative deterioration. We observed lower thiobarbituric acid-reactive substances (TBARS) in Gluteus medius of steers fed HTS before it was displayed (P=0.028), which could be explained by a reduced response to stress in these animals. While steers finished with corn and corn+HTS had elevated plasma cortisol at the end of the feeding period (P=0.047 and 0.093, respectively), animals fed HTS and corn+vitamin E did not. However, feeding HTS increased the rate of discoloration and TBARS accumulation after aerobic display of Longissimus lumborum and Gluteus medius. Diet did not affect the activity of oxidation-related enzymes and fatty acid composition of muscle. The accelerated rate of lipid oxidation during display of beef could be partially explained by a numerically lower concentration of tocopherols in the tissue.

Growth performance and muscle oxidation in rats fed increasing amounts of high-tannin sorghum.

Oxidative processes deteriorate the quality of meat products. High tannin sorghums (HTS) contain flavonoid oligomers known as proanthocyanidins or condensed tannins. These compounds act as anti-oxidants in vitro, but their effectiveness in vivo remains unclear. We tested the hypothesis that moderate amounts of dietary HTS could reduce markers of oxidation on muscle of rats without having detrimental effects in growth. We used 2 groups of 38 male Sprague Dawley rats at 5 and 13 wk of age each. Each age group was fed 4 diets in a completely randomized design. The younger group was fed the experimental diets for 10 wk (10W); whereas the older group was fed for 2 wk (2W). The diets were modified from the NIH-07 diet and contained HTS and corn at ratios of 0:50 (S0, control), 20:30 (S20), 35:15 (S35), and 50:0 (S50) as a percentage of the diet. Growth and the efficiency of gain were assessed periodically measuring BW, ADFI, ADG, and G:F. Oxidation in muscle was measured in fresh tissue and after 6 d of aerobic-refrigerated storage. Muscles evaluated were LM and soleus (SM). Fresh liver was also evaluated. Thiobarbituric acid-reactive substances (TBARS) and carbonyl content were used as markers of lipid and protein oxidation, respectively. No differences in BW, ADFI, ADG, and G:F were observed in 2W rats. Greater (P < 0.05) ADFI and ADG were observed in 10W-S35 group between d 1 and 7 and greater BW (P = 0.049) was observed in group 10W-S35 at d 70 compared with 10W-S0. No differences were observed between S0 and any HTS diet in G:F in 10W and 2W rats. No differences in TBARS or carbonyls were observed in liver. No differences in TBARS were observed in fresh and aged LM and SM. When LM samples were aged for 6 d, decreased carbonyl contents (P < 0.01) were observed in 10W-S35 and 10W-S50 diets compared with 10W-S0. Reductions in carbonyls were also observed in aged SM between 2W-S50 and 2W-S0 (P = 0.013). We concluded that inclusion of 35% HTS in the diet increased intake and growth rate of young, fast-growing rats without changing the efficiency of gain. Feeding HTS reduced markers of protein oxidation in rat muscle after 6 d of refrigerated storage. If similar results are observed in animals such as swine or cattle, the use of HTS as animal feed should be reassessed.

Quantification of plasma DNA as a prognostic indicator in canine lymphoid neoplasia.

Dogs have a similar incidence of spontaneous cancers as people, and a noninvasive test to monitor disease status in dogs would be of great value. Humans with cancer often have increased levels of cell-free circulating DNA in their plasma, which has shown promise for diagnosis, prognosis and detection of residual disease. We hypothesized that dogs with cancer have increased circulating DNA compared with healthy dogs or dogs with non-neoplastic diseases. Plasma DNA was measured in 40 healthy dogs, 20 dogs with non-neoplastic diseases and 80 dogs with cancer. The reference interval for plasma DNA in healthy dogs was 1-15 ng mL(-1). Dogs with lymphoma and lymphoid leukaemia had significantly higher concentrations (range: 0-91 ng mL(-1), P < 0.0001). Antigen receptor rearrangement assays suggest that plasma DNA had the same clonality as the primary lymphoid tumours. Dogs with lymphoid neoplasia and plasma DNA >25 ng mL(-1) had shorter remission times than those with < 25 ng mL(-1) (P = 0.0116). In contrast to humans, where increased plasma DNA is seen in many diseases, dogs with nonlymphoid malignancies and non-neoplastic diseases had plasma DNA concentrations similar to healthy dogs. This study shows that a portion of dogs with lymphoid neoplasia have increased tumour-derived plasma DNA, which serves as a negative prognostic indicator.

Detection and identification of bacteria using direct injection inductively coupled plasma mass spectroscopy.

In this study, an inductively coupled plasma mass spectrometer (ICP-MS) was used for the inorganic chemical characterization of biological materials by direct injection. ICP-MS has the advantage of sub-nanogram/gram detection limits for most elements making it a sensitive tool for the detection and characterization of aerosolized biological material. Suspended, microgram-sized samples of Bacillus subtilis spores (BG), Bacillus subtilis vegetative cells (Bg), and Bacillus thuringiensis (Bt) were analyzed via direct injection and they exhibit unique chemical signatures reflecting the processing history of each organism.

Shedding of Escherichia coli O157:H7 in dairy cattle housed in a confined environment following waterborne inoculation.

A study of Escherichia coli O157:H7 transmission and shedding was conducted with bull calves housed in individual pens within a confined environment. For comparative purposes, the numbers and duration of E. coli O157:H7 shedding in naturally infected calves were monitored after a single purchased calf (calf 156) tested positive prior to inoculation. During the next 8 days, the calves in adjacent pens and a pen directly across a walkway from calf 156 began to shed this serotype O157:H7 strain. Five of the eight calves in this room shed this O157:H7 strain at some time during the following 8 weeks. The numbers of E. coli O157:H7 isolates shed in these calves varied from 60 to 10(5) CFU/g of feces, and the duration of shedding ranged from 17 to >31 days. The genomic DNAs from isolates recovered from these calves were indistinguishable when compared by using XbaI digestion and pulsed-field gel electrophoresis. Inoculation of calves with 1 liter of water containing ca. 10(3) to 10(4) CFU of E. coli O157:H7/ml resulted in shedding in 10 of 12 calves (trial 1, 4 of 4 calves; trial 2, 6 of 8 calves). The inoculated calves shed the inoculation strain (FRIK 1275) as early as 24 h after administration. The duration of shedding varied from 18 to >43 days at levels from 10(2) to 10(6) CFU/g of feces. The numbers of doses necessary to initiate shedding varied among calves, and two calves in trial 2 never shed FRIK 1275 after four doses (ca. 10(6) CFU per dose). Results from this study confirm previous reports of animal-to-animal and waterborne dissemination of E. coli O157:H7 and highlight the need for an effective water treatment to reduce the spread of this pathogen in cattle.

The relative effect of dietary energy density on repleted and resting muscle glycogen concentrations.

The aim of this study was to assess the effect of dietary energy density on the glycogen concentration of bovine gluteus medius (GM) and longissimus thoracis et lumborum (LTL) muscles in castrated and intact males. Cattle received diet C (90% whole corn and supplement: 10% alfalfa haylage) or diet AH (100% alfalfa haylage) in three experiments involving a 2×2 factorial cross-over design. Cattle fed an AH diet for 70 days prior to the first experiment had high glycogen concentrations in the GM (129±9.9 mmol/kg) and LTL (108±7.6 mmol/kg) at the first biopsy session. These concentrations are similar to GM (133±6.2 mmol/kg) and LTL (105±5.8 mmol/kg) glycogen concentrations when these cattle subsequently received diet C for 30 or 37 days. Diet C increased muscle glycogen concentration, but the consistency of the increase in glycogen was dependent on the initial concentration. When the initial glycogen concentration was depleted to 50 µmol/g with one subcutaneous injection of adrenaline, repleted glycogen concentration was higher for diet C than AH (P<0.01) in all comparisons involving GM and LTL muscles of both steers and bulls. For resting muscle glycogen, the GM of bulls was more sensitive to the effect of diet C than the bull LTL or either muscle in steers. Diet was not a major determinant of resting muscle glycogen concentration. The ultimate pHs of GM, LTL and semimembranosus (SM) were not affected by diet and residual glycogen was lower in LTL of bulls fed either diet than for LTL of steers fed AH. Diet and male status did not affect residual glycogen in GM or SM.

Glycogen concentrations in bovine longissimus dorsi muscle.

The aim of this study was to assess the effect of location within bovine longissimus dorsi (thoracis et lumborum) muscle (LTL) when determining glycogen concentration. Six locations in Angus heifers were sampled post mortem (EXP-1) and eight locations in live Charolais crossbred steers were biopsied (EXP-2). In EXP-1, there was more glycogen in the animals' left LTL (64.1±1.6 mmol/kg) versus the right LTL (57.0±1.6 mmol/kg) (P<0.05). Locations along the cranial-caudal axis within LTL did not differ in glycogen concentration. Results for EXP-2 did not confirm the sidedness effect. Instead, it showed that glycogen concentration was lower at the cranial sampling locations near the 10th rib (89±2.5 mmol/kg) than at the middle (97±2.0 mmol/kg) or caudal locations (96±1.9 mmol/kg) (P<0.005). Taking lactate accumulation into account (glycolytic potential) rendered those differences insignificant. Nevertheless, the tendency remained clear. The potential for dilution of glycogen by intramuscular fat deposits is discussed. The results indicate that one needs to be cautious in performing repeated sampling on bovine LTL.

Effects of muscle α-tocopherol level and surface microbiological contamination on retail caselife of fresh beef from the US, Japan and Australia.

This study evaluated effects of dietary supplementation of vitamin E (1000 IU vitamin E/daily for 100 days prior to harvest) to fed cattle on retail caselife performance of fresh US beef in an export market (Japan). Economic performance (monetary losses associated with color deterioration) for US beef from vitamin E supplemented cattle vs beef from non-vitamin E supplemented cattle was contrasted. An additional, controlled study was performed to compare muscle α-tocopherol concentrations, color changes and microbiological growth for fresh beef derived from vitamin E supplemented US cattle and fresh beef from cattle with an unknown history, but from other countries. Australian strip loin steaks had the highest muscle α-tocopherol concentrations (4.6 µg/g tissue), followed by US strip loin steaks derived from vitamin E supplemented cattle (3.4 µg/g tissue) and Japanese strip loin steaks (2.8 or 2.5 µg/g tissue). US strip loin steaks from non-vitamin E supplemented cattle had the lowest (p<0.05) α-tocopherol levels (1.7 µg/g tissue). Aerobic plate counts and total coliform counts were generally low at 0 days of retail display, and they changed similarly among treatments over 6 days of display, regardless of the country of origin of the beef. Vitamin E supplementation of US cattle reduced total Japanese retail store losses due to discoloration of US beef, in yen, by 5.2 percentage points (p<0.05), saving Japanese retailers US $0.24/kg. Data suggest that US beef-normally perceived, in Japan, to discolor more quickly in the retail display case than beef from Australia-would compete more favorably, in shelf-life, with beef from other countries if it was derived from cattle that had been fed supplemental vitamin E.

Assessment of reductive acetogenesis with indigenous ruminal bacterium populations and Acetitomaculum ruminis.

The objective of this study was to evaluate the role of reductive acetogenesis as an alternative H2 disposal mechanism in the rumen. H2/CO2-supported acetogenic ruminal bacteria were enumerated by using a selective inhibitor of methanogenesis, 2-bromoethanesulfonic acid (BES). Acetogenic bacteria ranged in density from 2.5 x 10(5) cells/ml in beef cows fed a high-forage diet to 75 cells/ml in finishing steers fed a high-grain diet. Negligible endogenous acetogenic activity was demonstrated in incubations containing ruminal contents, NaH13CO3, and 100% H2 gas phase since [U-13C]acetate, as measured by mass spectroscopy, did not accumulate. Enhancement of acetogenesis was observed in these incubations when methanogenesis was inhibited by BES and/or by the addition of an axenic culture of the rumen acetogen Acetitomaculum ruminis 190A4 (10(7) CFU/ml). To assess the relative importance of population density and/or H2 concentration for reductive acetogenesis in ruminal contents, incubations as described above were performed under a 100% N2 gas phase. Both selective inhibition of methanogenesis and A. ruminis 190A4 fortification (>10(5) CFU/ml) were necessary for the detection of reductive acetogenesis under H2-limiting conditions. Under these conditions, H2 accumulated to 4, 800 ppm. In contrast, H2 accumulated to 400 ppm in incubations with active methanogenesis (without BES). These H2 concentrations correlated well with the pure culture H2 threshold concentrations determined for A. ruminis 190A4 (3,830 ppm) and the ruminal methanogen 10-16B (126 ppm). The data demonstrate that ruminal methanogenic bacteria limited reductive acetogenesis by lowering the H2 partial pressure below the level necessary for H2 utilization by A. ruminis 190A4.

Beef color update: the role for vitamin E.

Dietary supplementation of livestock with vitamin E results in improved quality of meat subsequently obtained from these animals. The effect is especially noteworthy in cattle, in which the primary effects are delayed discoloration and lipid oxidation. A threshold level of alpha-tocopherol in muscle ensures a detectable effect; dietary strategies for attaining this threshold must consider tocopherol status of cattle arriving at the feedyard and duration and level of supplementation. The alpha-tocopherol concentration in muscle must be determined before proper interpretation of experimental results can be made. Muscles vary in their color stability, and this relative difference is not changed by vitamin E supplementation. Several in vitro models have been used to characterize the interaction between alpha-tocopherol, lipid oxidation, and oxymyoglobin oxidation. Alpha-tocopherol seems to exert its color-stabilizing effect by indirectly delaying oxymyoglobin oxidation via direct inhibition of lipid oxidation. However, recent results demonstrating a protective effect of alpha-tocopherol toward oxymyoglobin in low-oxygen atmospheres indicate that additional mechanisms may exist. A better understanding of the fundamental bases for protection of water-soluble myoglobin by lipid-soluble alpha-tocopherol is needed to optimize this beneficial effect.

Transfer of fresh and cryopreserved IVP bovine embryos: normal calving, birth weight and gestation lengths.

In vitro and in vivo developmental competence of fresh and cryopreserved in vitro produced (IVP) bovine embryos was evaluated up to birth. Three experiments were done. The objective in the first experiment was to develop an optimal vitrification procedure for IVP bovine embryos by determining effects of exposure time (2, 5, 10, 20 min) and temperature (4, 22, 27 degrees C) in cryoprotective agents prior to vitrification on their post-thaw viability. The best combination was used in Experiments 2 and 3. In the second experiment, the importance of post-thaw morphologic selection on pregnancy rates was determined by transferring either selected or unselected single embryos. In the third experiment, pregnancy initiation, maintenance and calving results of vitrified embryos were compared with fresh and conventionally frozen embryos. Fetal losses, birth weights, gestation lengths and frequency of dystocia in the third experiment were monitored. The interaction of exposure time and temperature on both post-thaw re-expansion and hatching rates was significant (P < 0.01). Five minute exposure at 27 degrees C was optimal. In the second experiment, post-thaw selected vitrified embryos had higher pregnancy rates than unselected embryos (P < 0.05). In the third experiment, the pregnancy rate of vitrified embryos did not differ from that of fresh embryos (P > 0.05). However, pregnancy rate of conventionally frozen embryos was lower than that of fresh or vitrified embryos (P < 0.05). Of 92 calves born, 53 were male and 39 were female. Birth weights and dystocia scores of single-born calves did not differ between sexes (P > 0.05). Twin-born calves were lighter than single-born calves (P < 0.05). Overall, the data demonstrate that the transfer of vitrified IVP bovine embryos can result in healthy, apparently normal calves similar to those derived from transfer of fresh and conventionally frozen IVP bovine embryos.

Normal calves from transfer of biopsied, sexed and vitrified IVP bovine embryos.

Data on biopsied, sexed and cryopreserved in vitro produced (IVP) bovine embryos, and their in vivo developmental competence are very limited. Two preliminary studies were conducted before the primary study. In Experiment 1, post-thaw in vitro developmental competence of biopsied and vitrified IVP embryos was evaluated using re-expansion as an endpoint. In Experiment 2, the pregnancy rates of biopsied fresh, frozen or vitrified embryos following single embryo transfer were compared. Since vitrified embryos resulted in a higher pregnancy rate than frozen-thawed embryos, in the primary study (Experiment 3), all IVP embryos were vitrified following biopsy and sexing (by DNA fingerprinting). In Experiment 3, we compared pregnancy initiation and calving results of heifers in the following treatments: 1) artificial insemination (AI); 2) AI plus contralateral transfer of a single embryo (AI + SET); 3) ipsilateral transfer of single embryo (SET); or 4) bilateral transfer of two embryos (DET). Birth weights, gestation lengths and dystocia scores were recorded. In Experiment 1, post-thaw re-expansion rate of biopsied and vitrified embryos was 85% (70/82). In Experiment 2, pregnancy rates (90 d) were 44% (7/16), 23% (3/13), and 50% (7/14) for vitrified, frozen and fresh embryos, respectively (P < 0.10). In Experiment 3, pregnancy rates of AI and SET were 65% (20/31) and 40% (16/40), respectively (P < 0.05). The pregnancy rate of AI + SET was 75% (27/36) with 11 carrying twins, and the pregnancy rate of DET was 72% (26/36) with 10 carrying twins. All AI fetuses were carried to term, but only half the SET fetuses were carried to term. Similar calving rates were observed in the AI + SET and DET groups, 76 and 70%, respectively, of those pregnant at Day 40. Mean birth weight, dystocia score and gestation length of AI calves were not different from those of SET calves. Mean birth weight and dystocia score of single-born calves were greater than those of twin born calves (P < 0.05). These data demonstrate that biopsied IVP bovine embryos can be successfully cryopreserved by vitrification and following post-thaw embryo transfer, acceptable rates of offspring with normal birth weights can be obtained without major calving difficulties.

Tapeworm infection decreases intestinal transit and enteric aerobic bacterial populations.

Intestinal myoelectric patterns in rats are altered after chronic luminal infection with the tapeworm Hymenolepis diminuta. This study evaluates whether these altered patterns were associated with changes in intestinal fluid transit and endogenous enteric microbe levels. Luminal transit, measured throughout the small intestine during the interdigestive state, was significantly decreased during tapeworm infection. Reduced transit was regional, occurring in the same location as that of the tapeworm and maximal myoelectric alterations. In other experimental systems, aerobic and anaerobic bacterial overgrowth is associated with decreased transit; however, reduced transit during tapeworm infection was unexpectedly associated with decreased numbers of aerobic bacteria, whereas anaerobic bacterial populations remained unchanged. The lack of overgrowing endogenous microflora suggests that overgrowth is not responsible for tapeworm-stimulated alterations in host myoelectric patterns. We speculate that a tapeworm secretion could be responsible for both transit and motility changes while delayed intestinal transit could prevent tapeworm expulsion, aid the tapeworms' migration, and contribute to the digestion and absorption of nutrients by hosts and/or parasites.

Technical note: a simplified procedure for vitamin E determination in beef muscle.

A simplified method for the determination of alpha-tocopherol concentration in beef muscle was developed and evaluated. The method consists of a saponification step applied to 1-g samples of intact, fresh muscle, followed by a single isooctane extraction of the saponified samples. alpha-Tocopherol in the extract was separated by normal phase chromatography and quantified by fluorescence detection. A single extraction with the simplified method accounted for 95% of the total muscle alpha-tocopherol concentration obtained by two extractions with the Arnold et al. (J. Food Sci. 58:28, 1993) method. Recovery of added alpha-tocopherol standard after two extractions of a saponified muscle sample was 91% for the simplified method, which was not different (alpha = .78) from recovery using the Arnold method, and the efficiency of the single extraction in the simplified method was 89%. The coefficients of variation for the simplified and Arnold methods were both 3.1%. This method should permit the duplicate analysis of 100 fresh muscle samples within 24 h.

Growth and fermentation responses of Selenomonas ruminantium to limiting and non-limiting concentrations of ammonium chloride.

The objective of this study was to assess fermentation product, growth rate and growth yield responses of Selenomonas ruminantium HD4 to limiting and non-limiting ammonia concentrations. The ammonia half-inhibition constant for S. ruminantium in batch culture was 296 mM. Cells were grown in continuous culture with a defined ascorbate-reduced basal medium containing either 0.5, 5, 25, 50, 100 or 200 mM NH4Cl and dilution rates were 0.07, 0.14, 0.24 or 0.40 h-1. Ammonia was the growth-limiting nutrient when 0.5 mM NH4Cl was provided and the half-saturation constant was 72 microM. Specific rates of glucose utilization and fermentation acid carbon formation were highest for 0.5 mM NH4Cl. Lactate production (moles per mole of glucose disappearing) increased at the fastest dilution rate (0.40 h-1) for 5.0 mM NH4Cl while acetate and propionate decreased when compared to slower dilutions (0.07 and 0.14 h-1). Lactate production remained low while acetate and propionate remained high for all dilution rates when NH4Cl concentrations were 25 mM or greater. Yield (Y(Glc) and Y(ATP)) were nearly doubled when NH4Cl was increased from 0.5 mM (25.1 g cells/mol glucose used and 13.9 g cells/mol ATP produced respectively) to the higher concentrations. Y(Glc) was highest at 25 mM and 50 mM NH4Cl (48.2 cells/mol and 43.1 cells/mol respectively) as was Y(ATP) (23.2 cells/mol and 20.8 cells/mol respectively). Y(NH3) was highest at the lowest NH4Cl concentration. The maximal fermentation product formation rate occurred at a growth-limiting ammonia concentration, while maximal glucose and ATP bacterial yields occurred at non-growth-limiting ammonia concentrations. Given the growth response of this ruminal bacterium, it is possible that maximization of ruminal bacterial yield may necessitate sacrificing the substrate degradation rate and vice versa.

Color coordinates for assessment of dietary vitamin E effects on beef color stability.

Color stability was investigated in longissimus lumborum (LL), semimembranosus (SM), and gluteus medius (GM) muscles from Holstein steers fed diets including doses of alpha-tocopheryl acetate that were 0 (EO), 250 (E250), 500 (E500), and 2,000 (E2000) mg.steer-1.d-1 for 42 or 126 d. Longissimus lumborum was aged for 14, 28, and 56 d and GM and SM were aged for 14 d. Effects of vitamin E dose on retention of redness (a*), yellowness (b*), color saturation (chroma), and proportions of redness and yellowness (hue angle) following an aging period of 14 d were E2000 > E500 = E250 > EO (P < .01). Effectiveness of dose duration on the color parameters was 126 d > 42 d (P < .01). Dietary vitamin E supplementation stabilized redness and color saturation, decreased yellowness, and extended color display life of fresh beef. A technique for estimation of color display life based on hue angle measurements of fresh beef is described. Color display life estimates based on hue angle measurements were more consistent with vitamin E supplementation effects on metmyoglobin percentage and hue angle than were estimates obtained from the metmyoglobin threshold method. Color display life across LL, SM, and GM stored until d 14 and then displayed under simulated retail conditions was extended (P < .01) 2.0 (E250) to 5.0 d (E2000). Coefficients of determination for regressions of color display life on muscle alpha-tocopherol concentration were .81, .64, and .63 in LL, SM, and GM muscles aged 14 d, respectively. Supplementation of 500 mg of alpha-tocopheryl acetate per steer daily improved (P < .01) the mean color display life of these three muscles by 2.3 d, or 100%.