Combinations of bacterial cultures, exogenous enzymes, and yeast-based feed additives and their impact on ruminal microbiome.

Our objective was to evaluate the effects of bacteria (Lactobacillus animalis, Propionibacterium freudenreichii, Bacillus lichenformis, Bacillus subtilis, and Enterococcus faecium), enzymes (amylase, hemicellulose, and xylanase), and yeast as additives on the ruminal microbiome. We hypothesized that inclusion of bacteria, enzymes, and yeast would impact butyric bacterial populations. Eight fermenters were arranged in a duplicated 4 × 4 Latin square with the following treatments: 1) control without additives (CTRL); 2) bacterial culture and enzyme blend (EB); 3) bacterial culture and enzyme blend with a live yeast and yeast culture blend (EBY); and 4) double dose of bacterial culture and enzyme blend and the yeast products blend (2X). We conducted four fermentation periods of 10 d each, with the last 3 d for collection of samples. Overall, 64 solid and liquid samples were analyzed by amplification of the V4 region of bacterial 16S rRNA. Data were analyzed with R and SAS. The following orthogonal contrasts were used: 1) ADD-the control compared to all treatments with additives (CTRL vs. EB, EBY, and 2X); 2) YEAST-treatment without yeast compared to those with yeast (EB vs. EBY and 2X); and 3) DOSE-the single dose of enzymes, bacteria, and yeast compared to the doubled dose (EBY vs. 2X). Family Prevotellaceae was more abundant when additives were added (ADD). Additives (ADD) also increased relative abundance of Prevotellaceae Ga6A1 and YAB2003 in solid fraction, and of Prevotellaceae Ga6A1 and two members of Lachnospiracea family in liquid fraction. Yeast (YEAST) decreased relative abundance of Succinivibrionaceae UCG-001 and increased abundance of Ruminococcus and Prevotellaceae UCG-003 in solid fraction. Doubling the dose of enzymes and microbial additives (DOSE) decreased the abundance of Succiniclasticum in solid fraction and Selenomonadaceae in the liquid. Molar proportion of butyrate was highly correlated with abundance of Prevotellaceae Ga6A1 in solid (r = 0.68) and liquid fraction (r = 0.79), and with Unclassified Lachnospiraceae in liquid (r = 0.70). Our results demonstrate that YEAST decreases abundance of succinate synthesizing bacteria, while DOSE decreases abundance of bacteria that metabolize succinate into propionate. Combined bacteria, enzymes, and yeast increase the relative abundance of specific genera primarily within the Prevotellaceae family, which may explain the increase in butyrate molar proportion observed with ADD.

Effects of bacterial cultures, enzymes, and yeast-based feed additive combinations on ruminal fermentation in a dual-flow continuous culture system.

Bacterial cultures, enzymes, and yeast-derived feed additives are often included in commercial dairy rations due to their effects on ruminal fermentation. However, the effects of these additives when fed together are not well understood. The objective of this study was to evaluate the changes in ruminal fermentation when a dairy ration is supplemented with combinations of bacterial probiotics, enzymes and yeast. Our hypotheses were that ruminal fermentation would be altered, indicated through changes in volatile fatty acid profile and nutrient digestibility, with the inclusion of (1) an additive, (2) yeast, and (3) increasing additive doses. Treatments were randomly assigned to 8 fermenters in a replicated 4 × 4 Latin square with four 10 d experimental periods, consisting of 7 d for diet adaptation and 3 d for sample collection. Basal diets contained 52:48 forage:concentrate and fermenters were fed 106 g of dry matter per day divided equally between two feeding times. Treatments were: control (CTRL, without additives); bacterial culture/enzyme blend (EB, 1.7 mg/d); bacterial culture/enzyme blend with a blend of live yeast and yeast culture (EBY, 49.76 mg/d); and a double dose of the EBY treatment (2×, 99.53 mg/d). The bacterial culture/enzyme blend contained five strains of probiotics (Lactobacillus animalis, Propionibacterium freudenreichii, Bacillus lichenformis, Bacillus subtilis, and Enterococcus faecium) and three enzymes (amylase, hemicellulase, and xylanase). On d 8-10, samples were collected for pH, redox, volatile fatty acids, lactate, ammonia N, and digestibility measurements. Statistical analysis was performed using the GLIMMIX procedure of SAS. Repeated measures were used for pH, redox, VFA, NH3-N, and lactate kinetics data. Orthogonal contrasts were used to test the effect of (1) additives, ADD (CTRL vs. EB, EBY, and 2X); (2) yeast, YEAST (EB vs. EBY, and 2X); and (3) dose, DOSE (EBY vs. 2X). No effects (P > 0.05) were observed for pH, redox, NH3-N, acetate, isobutyrate, valerate, total VFA, acetate:propionate, nutrient digestibility or N utilization. Within the 24 h pool, the molar proportion of butyrate increased (P = 0.03) with the inclusion of additives when compared to the control while the molar proportion of propionate tended to decrease (P = 0.07). In conclusion, the inclusion of bacterial cultures, enzymes and yeast in the diet increased butyrate concentration; but did not result in major changes in ruminal fermentation.

Effects of calcium-magnesium carbonate and calcium-magnesium hydroxide as supplemental sources of magnesium on microbial fermentation in a dual-flow continuous culture.

Supplemental sources of Mg can also aid in ruminal pH regulation due to their alkaline properties. Magnesium oxide (MgO) is the most common source of Mg for ruminants and can help controlling ruminal pH; however, the alkaline potential of other sources of Mg has not been evaluated. We aimed to evaluate the inclusion of calcium-magnesium carbonate (CaMg(CO3)2) and calcium-magnesium hydroxide (CaMg(OH)4) alone or in combination as supplemental sources of Mg in corn silage-based diets and its impact on ruminal microbial fermentation. We hypothesized that inclusion of CaMg(OH)4 would allow for ruminal fermentation conditions resulting in a greater pH compared to the inclusion of CaMg(CO3)2. Four treatments were defined by the supplemental source of Mg in the diet: 1) Control (100% MgO, plus sodium sesquicarbonate as a buffer); 2) CO3 [100% CaMg(CO3)2]; 3) OH [100% CaMg(OH)4]; and 4) CO3/OH [50% Mg from CaMg(CO3)2, 50% Mg from CaMg(OH)4]. Nutrient concentration was held constant across treatments (16% CP, 30% NDF, 1.66 Mcal NEl/kg, 0.67% Ca, and 0.21% Mg). Four fermenters were used in a 4 × 4 Latin square design with four periods of 10 d each. Samples were collected for analyses of nutrient digestibility, soluble Mg, VFA, and NH3, while pH was measured at 0, 1, 2, 4, 6, 8, and 10 h post morning feeding to estimate % time when pH was below 6 (pH-B6) and area under the pH curve for pH below 6.0 (pH-AUC). Bacteria pellets were harvested for 15N analysis and estimates of N metabolism. Treatment effects were analyzed with the mixed procedure of SAS, while effects of using either CaMg(CO3)2 or CaMg(OH)4 as Mg source in comparison to Control treatment were evaluated by orthogonal contrasts. Similar pH-related variables were observed for Control, OH, and CO3/OH treatments, which had smaller pH-AUC and pH-B6 than CO3 (P ≤ 0.01). Butyrate molar proportion was greater in Control and CO3/OH than in CO3 and OH (P = 0.04). Orthogonal contrasts showed lower flow of bacterial N (P = 0.04), lower butyrate molar proportion (P = 0.08) and greater pH-AUC (P = 0.05) for diets with CaMg(CO3)2 in comparison with the Control. Concentration of soluble Mg in ruminal fluid (P = 0.73) and nutrient digestibility (P ≥ 0.52) were similar across treatments. Under the conditions of this experiment, using CaMg(OH)4 alone or combined with CaMg(CO3)2 allowed for a less acidic ruminal fermentation pattern than a diet with only CaMg(CO3)2.

Right-sided congestive heart failure in a dog because of a primary intracavitary myocardial lymphoma.

CASE REPORT: A 9-year-old female intact Cocker Spaniel was presented with a history of acute-onset dyspnoea and abdominal distension of 3 days' duration. Ultrasonography revealed pleural, peritoneal and pericardial effusions. Abdominal fluid analysis was consistent with a modified transudate. Echocardiography revealed a large, hypoechoic space-occupying mass within the right atrium. The dog was euthanased and the postmortem examination showed a solid, 40 × 35 × 20 mm broad-based mass arising from the right atrial wall and occluding approximately 90% of the right atrial lumen. Histopathology revealed myocardial lymphoma. There were histologically similar, focal nodules in the lung parenchyma without involvement of other extracardiac sites. There was gross and histological evidence of hepatic congestion and marked distension of the caudal vena cava, consistent with secondary right-sided congestive heart failure. CONCLUSION: This case highlights the need to consider lymphoma as a differential diagnosis for an intra-atrial mass and as a cause of congestive heart failure in the dog.

Gallbladder mucocoele and concurrent hepatic lipidosis in a cat.

A 3-year-old Domestic Shorthair cat was presented with weight loss, anorexia and icterus. Feline hepatic lipidosis and gallbladder mucocoele were diagnosed; this is the first report of gallbladder mucocoele in the cat. The case was managed successfully with cholecystojejunostomy, gastrostomy tube placement and tube feeding for 3 months. The cat has survived over the long term with minimal complications.

Photosensitisation induced by clofazimine in a cat.

A pale ginger cat was treated with clofazimine for feline leprosy. During the course of treatment, photosensitisation by clofazimine led to a dermatological disease resembling actinic dermatitis. The mycobacterial disease was eventually cured with clofazimine and the photosensitisation was managed by regulating sunlight exposure. Upon review of the literature, this appears to be the first documented case of drug-induced photosensitisation in the cat.

Reference limits for urinary fractional excretion of electrolytes in adult non-racing Greyhound dogs.

OBJECTIVE: To determine reference limits for urinary fractional excretion of electrolytes in Greyhound dogs. METHODS: Urinary fractional excretion was calculated using a spot clearance method preceded by a 16 to 20 hour fast in 48 Greyhound dogs. Raw data analysed using the bootstrap estimate was used to calculate the reference limits. RESULTS: The observed range for urinary fractional excretion in Greyhound dogs was 0.0 to 0.77% for sodium, 0.9 to 14.7% for potassium, 0 to 0.66% for chloride, 0.03 to 0.22% for calcium and 0.4 to 20.1% for phosphate. Expressed as percentages, the suggested reference limits for fractional excretion in Greyhound dogs are as follows: sodium < or = 0.72, potassium < or = 12.2, chloride < or = 0.55, calcium < or = 0.13 and phosphate < or = 16.5. CLINICAL SIGNIFICANCE: Veterinary practitioners may use these reference limits for urinary electrolyte fractional excretion when investigating renal tubular disease in Greyhound dogs.

Lymphoma(s) showing epitheliotropism and diffuse skeletal muscle involvement presenting as a polymyopathy in a young dog.

A 3-year-old dog presented with a severe polymyopathy and subsequently developed nodular skin lesions. The disease in this dog was caused by lymphoma, showing cutaneous epitheliotropism as well as infiltration of skeletal muscle in conjunction with polymyositis.

Balloon dilation of a urethral stricture in a dog.

A 2-year-old male neutered Scottish Terrier presented with dysuria caused by complete urethral obstruction due to a stricture secondary to catheter-induced trauma. The urethral obstruction was resolved by balloon dilation of the stricture under fluoroscopic guidance.

Effects of electrode montage on the spectral composition of the infant auditory brainstem response.

We evaluated the effect of electrode montage on the spectrum of the infant auditory brainstem response (ABR). Spectral profiles of ipsilateral, contralateral, noncephalic, and horizontal recordings obtained in response to slow and fast repetition rates and at low and high stimulus intensities were also evaluated. Findings indicate that the spectrum of the infant ABR is dominated by low-frequency energy, maximal below 200 Hz. The spectra of ipsilateral and noncephalic recordings are the strongest irrespective of stimulus intensity or repetition rate. Increase in stimulus intensity or repetition rate typically enhances the amount of energy below 200 Hz. These results reinforce the clinical utility of ipsilateral and noncephalic recordings for screening, threshold measurement, and neurodiagnostic purposes.

Effects of electrode montage on infant auditory brainstem response.

The present study compares ipsilateral, contralateral, horizontal and non-cephalic auditory-evoked brainstem responses (ABR) obtained from normal infants (39-42 weeks' post-conceptional age) to assess their utility in auditory neurodiagnosis. Wave latencies, wave amplitudes and waveform morphology were evaluated at slow and fast repetition rates. Ipsilateral and horizontal recordings obtained with a fast stimulation rate provide best waveform characteristics for neurodiagnostic interpretation. Further enhancement of wave V amplitude and separation of IV-V wave complex may be achieved by using the non-cephalic recording montage. Compromised or low amplitude components on contralateral measurements may confound neurological interpretation.

The ferning and nitrazine tests of amniotic fluid between 12 and 41 weeks gestation.

Most studies investigating the usefulness of ferning and nitrazine methods for detecting the presence of amniotic fluid following ruptured amniotic membranes have employed samples from vaginal pooling. Reported accuracies are 84 to 100% for the ferning test and 87 to 97% for the nitrazine test. Our study was designed to determine if these tests are applicable throughout gestation. Samples were obtained from amniocenteses between 14 and 42 weeks. Part I: Of 112 samples allowed to dry on a slide for 3 minutes only, 86.6% were ferning positive and 100% were nitrazine positive. Flame-drying increased the presence of ferning to 96.4%. Part II: 363 samples were allowed to dry completely for up to 10 minutes. All samples were ferning and nitrazine positive. Part III: 17 samples were tested for ferning and nitrazine at 1 hour, and again at 1 week. Sixty-six samples were tested within 1 week and again at 2 weeks. All samples remained ferning and nitrazine positive. Therefore optimal analysis of suspected amniotic fluid occurs when specimens are air-dried for 10 minutes. These tests may be reliably performed at gestations of 12 to 41 weeks.

Case report: survival of an infant with a birthweight of 345 grams.

Infant survival with ever-decreasing birthweights is attainable with recent advances in maternal-fetal medicine and enhanced neonatal intensive care. We report a gestation complicated by severe chronic hypertension and fetal distress necessitating delivery at 26 1/7 weeks. The growth-retarded newborn weighted 345 g (12 oz) and survived with minimal sequelae despite a protracted and complicated neonatal course.

Effect of penetration enhancers on the permeation of mannitol, hydrocortisone and progesterone through human skin.

Mannitol, hydrocortisone and progesterone were selected as model penetrants to assess the mode of action of eight potential penetration enhancers in human skin. Their partition coefficients, octanol: water and stratum corneum: water were measured and correlated with their postulated routes of penetration through human skin. The results suggest that mannitol penetrated via a polar route, hydrocortisone by a mainly lipid route and progesterone via a lipid pathway but its penetration rate was probably affected by aqueous layers. From permeation studies through cadaver skin in which an in-vivo mimic method was used, it was concluded that the penetration enhancers fell into three main categories: solvents which enhanced permeation of polar and non-polar compounds e.g. 2-pyrrolidone, N-methylpyrrolidone, N-methylformamide and propylene glycol plus Azone; enhancers which preferentially affected the polar route e.g. propylene glycol plus decylmethylsulphoxide, and accelerants which mainly modified the non-polar route e.g. propylene glycol plus oleic acid, propylene glycol alone and, to a limited extent, water.

Optimization of bioavailability of topical steroids: non-occluded penetration enhancers under thermodynamic control.

The non-occluded vasoconstrictor test under thermodynamic control assessed the effect of penetration enhancers on the topical bioavailability of a model steroid betamethasone 17-benzoate, using aqueous dimethylisosorbide (DMI) as a standard solvent. The aqueous potential penetration enhancers used were at 10% steroid saturation i.e. ideally at identical steroid thermodynamic activity. In the vasoconstrictor test, 2-pyrrolidone, N-methyl-2-pyrrolidone, propylene glycol with oleic acid, propylene glycol with azone and dimethylformamide (DMF) increased the steroid bioavailability compared with that from DMI, while propylene glycol alone produced borderline improvement. Azone and oleic acid in combination with DMI or Betnovate cream did not increase the steroid bioavailability indicating the importance of the correct cosolvent. The pyrrolidones established superior stratum corneum reservoirs compared with DMI, the other vehicles being similar to DMI. It was concluded that excepting DMI, the solvents tested were penetration enhancers for the model steroid betamethasone 17-benzoate and are worthy of further study. However, irritant effects may make some of them unacceptable for clinical use.