Effect of ractopamine on insulin sensitivity and response of isolated rat adipocytes

In vitro effects of the phenethanolamine ractopamine on basal and insulin-stimulated lipid metabolism were determined in adipocytes isolated from epididymal fat pads of Sprague-Dawley rats. Ractopamine appeared to be equipotent to the catecholamine isoproterenol in stimulating basal lipolysis and inhibiting basal lipogenesis, producing maximum effects at 10(-6) M. Addition of a half-maximally stimulating dose of ractopamine (5 x 10(-8) M) to the incubation media decreased insulin sensitivity but not insulin responsiveness of the cells, stimulating lipolysis and inhibiting lipogenesis only in the presence of low media insulin concentrations. This effect was totally reversed by 10 microM/propranolol. Maximally effective concentrations of ractopamine (10(-6) M) significantly decreased both the sensitivity and responsiveness of the isolated adipocytes to insulin. Addition of 10 microM propranolol to the incubation media effectively reversed the lipolytic and anti-lipogenic effects of 10(-6) M ractopamine observed at media insulin concentrations greater than 25 microU/ml, whereas it only partially reduced the ractopamine-induced effects observed at lower insulin concentrations. The results demonstrate 1) that ractopamine has concentration-dependent effects on adipose tissue insulin sensitivity and responsiveness and 2) that these effects may be mediated, in part, through beta-adrenergic receptors.     

Enterotoxin production at 4, 22, and 37 degrees C by Yersinia enterocolitica and Yersinia enterocolitica-like bacteria isolated from porcine tonsils and pork products

Altogether, 71 strains of Yersinia enterocolitica and Yersinia enterocolitica-like bacteria from porcine tonsils and pork products were examined for their ability to produce enterotoxin using the infant mouse assay. Of these, 37 strains (52.1 %) produced enterotoxin at 22 °C, 3 were positive at 4 and 22 °C, and 1 was enterotoxigenic at 22 and 37°C. No strain was positive at all 3 temperatures. The highest prevalence of enterotoxin production at 22°C was detected in serotype 0:11 (80.0%), followed by 0:3/ biotype 4 (74.2 %), and 0:12 (66.7 %). Enterotoxin production at 4°C was recorded in 2 (15.4 %) of the Yersinia kristensenii strains (0:11, 0:12) and 1 of the Yersinia enterocolitica strains, (0:3) examined. One Yersinia kristensenii strain (0:11) was enterotoxigenic at 37 °C. The results indicate that enterotoxin production is a common feature of yersiniae isolated from porcine tonsils and pork products in Norway and may represent a possible source of food borne intoxication.     

The density of equine tissue at 37 degrees C.

The density of the following equine tissues was measured: white and grey brain matter, myocardium, lung parenchyma, liver, spleen, gastrointestinal tract and contents, renal medulla and cortex, muscle, fat, tendon and bone. Statistical analysis indicated that there were highly significant differences in the values for density between horses and between tissues within horses. Values for density of eight different muscles were obtained for 18 horses and these were shown to differ highly significantly between horses and between muscles within horses. The values for splenic density differed highly significantly between those for knackery killed horses and for the barbiturate killed post morten room horses.     

Decreased insulin binding to porcine adipocytes in vitro by beta-adrenergic agonists

The present study was conducted to determine the influence of dibutyryl-cAMP (dbcAMP), epinephrine, ractopamine and clenbuterol on insulin binding to porcine adipocytes. Dibutyryl-cAMP decreased insulin binding to swine adipocytes by 40 and 20% at 1.8 and 25.8 ng insulin/ml, respectively. Ractopamine and clenbuterol directly reduced insulin binding at the low insulin concentration and decreased binding at high insulin concentrations in the presence of adenosine deaminase. Scatchard analysis suggested that the reduction of insulin binding was due to a decrease in receptor number. Epinephrine alone did not influence insulin binding. In the presence of theophylline, epinephrine decreased binding at both low and high insulin concentrations; however, ractopamine plus theophylline decreased binding only at the low insulin concentration. Clenbuterol did not affect insulin binding in the presence of theophylline. Propranolol blocked the inhibitory effect of epinephrine on insulin binding. These beta-adrenergic agonists can inhibit insulin binding and, thus, antagonize insulin action in swine adipocytes.     

Binding characteristics of swine erythrocyte insulin receptors

Crossbred gilts (controls; n = 7) had 8.8 +/- 1.1% (mean +/- SEM) maximum binding of [125I]insulin to insulin receptors on erythrocytes. The number of insulin-binding sites per cell was 137 +/- 19, with a binding affinity ranging from 7.4 X 10(7)M-1 to 11.2 X 10(7)M-1 and mean of 8.8 X 10(7)M-1. Pregnant sows (n = 5) had a significant increase (P less than 0.01) in maximum binding due to an increase in number of receptor sites per cell. Lactating sows fed a high-fiber diet (n = 3) and a low-fiber diet (n = 4) did not develop a significant difference in maximum binding of insulin. Sows fed the low-fiber diet had a significantly higher number of binding sites and a significantly lower binding affinity than did sows fed a high-fiber diet. Receptor-binding affinity was lower in the low-fiber diet group than in cycling gilts, whereas data from sows fed the high-fiber diet did not differ from data for cycling gilts. Data from this study indicated that insulin receptors of swine erythrocytes have binding characteristics similar to those in other species. Pregnancy and diet will alter insulin receptor binding in swine.     

Sensitivity of lipolysis and lipogenesis to dibutyryl-cAMP and beta-adrenergic agonists in swine adipocytes in vitro

The sensitivities of lipolysis and fatty acid synthesis to dibutyryl-cAMP (dbcAMP), epinephrine, ractopamine and clenbuterol were quantified in vitro using porcine adipocytes. Insulin-stimulated lipogenesis showed a biphasic response to dbcAMP, with increased rates at low concentrations and decreased (55%) rates at higher concentrations of dbcAMP. In the absence of insulin, lipogenesis was inhibited 78% by dbcAMP. In the presence of adenosine deaminase or theophylline, all three beta-adrenergic agonists inhibited basal lipogenesis, but only epinephrine and ractopamine inhibited insulin-stimulated lipogenesis. The relationship between suppressed lipogenesis and enhanced lipolysis in response to dbcAMP and the beta-agonists revealed that 1) basal lipogenesis was more sensitive to inhibition than was the stimulation of lipolysis, 2) sensitivity differences were magnified if adenosine deaminase was present and 3) insulin decreased adipocyte sensitivity to the inhibitory effects of dbcAMP and the beta-adrenergic agonists. These results indicate that the relative sensitivities of lipogenesis and lipolysis to beta-adrenergic stimulation can be modified by adenosine and insulin. Furthermore, adenosine and insulin antagonize beta-adrenergic responses, in part, by cAMP-independent mechanisms.     

Stimulation of lipogenesis by insulin in swine adipose tissue: antagonism by porcine growth hormone

The effects of physiological (1, 10 ng/ml) and pharmacological (1,000 ng/ml) concentrations of insulin (INS) and porcine growth hormone (pGH) on lipid metabolism were determined in short-term (2 h) and long-term (26, 50 h) incubations of swine adipose tissue. The short-term effects of three different commercial sources of bovine serum albumin (BSA) on adipose tissue metabolism were also evaluated. Two of the three BSA preparations were found to be unsuitable for inclusion in the short-term incubation buffer because they caused a stimulation of lipid synthesis in adipose tissue and masked the stimulatory effects of insulin. Physiological concentrations of insulin stimulated glucose metabolism in 2-h incubations by 100% in adipose tissue from 80-kg swine. After a 26-h incubation period, INS maintained rates of glucose metabolism at levels comparable to maximally stimulated rates in fresh tissue. Insulin also enhanced glucose metabolism following 50-h incubations; however, rates were less than for 2- or 26-h incubations. Glucose metabolism was also stimulated in adipose tissue from 127-kg swine when incubated for 2 h with INS; however, INS responsiveness declined with increasing body weight. Lipogenesis and glucose oxidation were partially maintained by INS using tissue from the heavier swine. A pharmacological but not physiological concentration of pGH stimulated glucose metabolism in short-term incubations by 50% in adipose tissue from 80-kg swine, and by 10% in adipose tissue from 127-kg swine. Long-term culture of adipose tissue in the presence of pGH had no effect on glucose metabolism. Physiological levels of pGH directly antagonized the stimulation of glucose metabolism by INS in short- and long-term incubations. In summary, these results are the first to establish that swine adipose tissue is quite sensitive to insulin and that pGH directly antagonizes insulin action.     

Antimicrobial resistance of Salmonella isolated from finishing swine and the environment of 60 Alberta swine farms

The study objective was to describe and evaluate antimicrobial resistance profiles in Salmonella isolated from Alberta swine finishing farms. Salmonella isolates (n = 322) were obtained from 192 fecal and 84 environmental samples of the 60 Salmonella-positive swine finishing farms. Isolates were classified susceptible, intermediate or resistant based on NCCLS guidelines. More than half of the isolates (53.4%) were susceptible to all of the 18 antimicrobials in the testing panel. No resistance was observed to amikacin, amoxicillin/clavulanic acid, cefoxitin, ceftiofur, ceftriaxone, cephalothin, ciprofloxacin, imipenem or nalidixic acid. Less than 1% of isolates were resistant to apramycin, gentamicin and trimethoprim/sulfamethoxazole. Higher frequencies of resistance were observed for chloramphenicol (4.7%), ampicillin (7.8%), kanamycin (11.8%), sulfamethoxazole (21.1%), streptomycin (25.5%) and tetracycline (38.8%). Eleven Salmonella serovars had isolates with resistance to > or =3 antimicrobials. The most frequently resistant serovar was Salmonella Derby, with 27 (38.0%) isolates resistant to > or =3 antimicrobials, including resistance to five and six antimicrobials. An absence of resistance to cephalosporins and fluoroquniolones and a low proportion of isolates resistant to amikacin, amoxicillin/clavulanic acid, apramycin, gentamicin and trimethoprim/sulfamethoxazole are encouraging findings from public health and animal health perspectives. Frequent resistance observed for ampicillin, kanamycin, sulfamethoxazole, streptomycin and tetracycline, antimicrobials commonly used in veterinary medicine for decades, indicates an urgent need to utilize these antimicrobials more prudently if their benefits are to be preserved.     

Antimicrobial resistance of Actinobacillus pleuropneumoniae isolated from swine

The aim of this retrospective study was to evaluate the antimicrobial resistance rates and the trend in resistance of Actinobacillus pleuropneumoniae isolated from pigs in Italy from 1994 to 2009. A total of 992 A. pleuropneumoniae isolates were tested for their susceptibility to a panel of antimicrobial agents in a disk diffusion method. Resistance to 7 drugs (amoxicillin, amoxicillin/clavulanic acid, ampicillin, cefquinome, cotrimoxazole, penicillin G and tilmicosin) showed a significant increasing trend over the time, while for 2 drugs (gentamycin and marbofloxacin) a significant decrease was observed. Resistance to the remaining 14 antimicrobial agents tested did not change significantly over the study period. Most of the isolates retained high susceptibility to antimicrobials usually effective against A. pleuropneumoniae such as amphenicols, fluoroquinolones and ceftiofur. However, high rates of resistance were observed for potentiated sulfa drugs, tetracyclines and penicillins which are currently recommended antimicrobials for pig pleuropneumonia therapy. Our results suggest the importance of continued monitoring of A. pleuropneumoniae clinical isolates in order to choose the most appropriate treatment of infections and to control the increase of resistance to currently used antimicrobials.     

Keratin degradation by bacteria and fungi isolated from a poultry farm and plumage

1. Poultry processing generates a large quantity of feather waste. Feathers are a rich source of keratin and could be used as a feather meal in the feed industry if the keratin is degraded using suitable micro-organisms.

2. In this study, keratin-degrading micro-organisms were isolated from a poultry farm. The predominant organisms were identified as Bacillus subtilis, Aspergillus flavus, A. fumigatus and Trychophyton sp.

3. The isolates were inoculated into feather medium and observed for keratin degradation by measuring the protein content, free amino acids and change in pH.

4. During feather degradation by B. subtilis, the concentration of soluble protein released to the medium increased gradually and reached the maximum (433 µg/ml) during d 7 of incubation and the pH increased from the initial 6.9 to 8.4 on d 9 of incubation. Similarly, the maximum protein content of 414 µg/ml and pH of 8.5 was observed for A. fumigatus on d 21 of incubation.

5. B. subtilis and A. fumigatus showed almost the same level of keratinase activity.     

猪源乳酸杆菌对永生化猪小肠上皮细胞的黏附性能

以永生化猪小肠上皮细胞ZYM—SIEC02为体外模型,通过革兰染色法、平板计数法等检测发酵乳杆菌Y091231（Lactobacillus fermentation）、乳酸乳杆菌Y091221（Lactobacillus lactis）,嗜酸乳杆菌Y224031（Lactoba-cillusacidophilus）的黏附性能及对大肠杆菌C83921、致病性沙门菌、金黄色葡萄球菌CVCC2258的黏附抑制作用。结果显示,3株菌对永生化猪小肠上皮细胞的黏附性均较强,黏附指数平均为：Y091231（2903．25±83．33）个／100cells、Y091221（2320．37±81．02）个／100ceils、Y224031（I965．43±37．72）个／100cells,菌株之间差异不显著（P〉0．05）。黏附抑制试验结果显示,Y091231对大肠埃希菌C83921和金黄色葡萄球菌CVCC2258的黏附抑菌能力最强（（99．4±3．17）％和（98．0±2．31）％）,Y091221对沙门菌的抑菌能力最强（（97．1±2．04）％）,Y224031对大肠埃希菌C83921比对另外2种致病菌的黏附抑制能力高,但又显著低于Y091231和Y091221（P〈0．05）,表明不同种乳酸菌对致病菌的黏附抑制作用具有菌种特异性。