Cloning and Sequencing of cnf1 from Escherichia coli Incriminated in Mink and Bovine Colibacillosis

Colibacillosis is responsible for significant losses to the mink and cattle industries. Previous work in our laboratory and by others has suggested that possession of cnf1, the gene encoding cytotoxic necrotizing factor (CNF1), may contribute to the virulence of isolates of E. coli from mink and cattle. The cnf1 gene from E. coli isolated from a mink with colisepticaemia and a bovid with scours was amplified and cloned as a 3.5 kb fragment, and the fragment was sequenced. The cnf1 sequences from the mink and bovine isolates of E. coli were compared to each other and to cnf1 sequences of E. coli from urinary tract and diarrhoea-associated infections of humans. The difference was only 7 nucleotides between the cnf1 sequences of the mink and bovine isolates of E. coli, which translated into 7 differences in amino acids. The cnf1 sequence of the mink isolate of E. coli had 15 nucleotide differences from the cnf1 sequences of the human isolate of E. coli (GenBank X70670), which translated into 11 differences in amino acids between these proteins. The cnf1 sequence of the bovine isolate of E. coli had 14 nucleotide differences from the cnf1 sequence of the human isolate of E. coli (GenBank X70670), which translated into 10 differences in amino acids between these proteins. The highly conserved sequences of the amino acids of CNF1 proteins make them a promising target for detection and control of the CNF1-producing E. coli involved in disease among various host species.     

Prior experience, antioxidants, and mitochondrial cofactors improve cognitive function in aged beagles

Results of this study support the free-radical theory of aging and demonstrated that providing higher levels of vitamin E in food resulted in higher serum vitamin E concentrations and improved performance on landmark-discrimination tasks in aged dogs. Factors other than vitamin E also contributed to the response but remain undefined.     

A novel cognitive palatability assessment protocol for dogs

Assessment of canine palatability is important for both the pet food and pharmaceutical industries; however, the current palatability assessment protocols are limited in their utility. The most common technique, the two-pan test, does not control for the satiating effects of food and may not be useful for long-term palatability analysis because nutritional or caloric characteristics of the diets may interfere with the results. Furthermore, the large quantities of foods consumed may be detrimental to the health of animals that do not self-limit their food intake. The purpose of this study was to determine whether a cognitive protocol could be used to determine food palatability in dogs. Five beagle dogs were trained on a three-choice object-discrimination learning task. After establishing object preferences, the preferred object was associated with no reward, a second object was associated with the dog's normal laboratory diet (Purina Agribrands Canine Lab Chow No. 5006; Agribrands Purina Canada, Inc., Woodstock, ON, Canada), and the third object was associated with a commercial (Hill's P/D; Hill's Pet Nutrition Inc., Topeka, KS) diet. In the discrimination-training phase, dogs were trained until they learned to avoid the no-reward object. They were subsequently given an additional 20 test sessions, which were used to determine food preference. In the reversal phase, which involved reversal learning, the object-food associations were modified, such that the object that was previously associated with Hill's P/D diet was now associated with the normal laboratory diet and vice versa. Once the dogs learned to avoid the no-reward object, they were tested for an additional 20 sessions. All subjects learned to avoid the no-reward object during the initial learning, and the number of choices to the object associated with the Hill's P/D diet was greater than the number of choices to the objects associated with the dry laboratory diet (P < 0.05) and no reward (P < 0.05), indicating a strong preference for the Hill's P/D diet. The object preferences were reversed in only three of five dogs when the food-choice associations were reversed, although the two phases did not differ significantly from one another. The protocol in the present study provides a robust measure of food palatability and circumvents many of the limitations associated with other palatability assessment techniques. The present protocol should be useful as a replacement or adjunct to other tests of palatability, but requires further validation by comparing the assessment of more similar and novel foods directly with other palatability tests.     

澳大利亚棉铃虫杀虫剂抗性的治理策略

澳大利亚棉铃虫杀虫剂抗性的治理策略ＮｅｉｌＷ．Ｆｏｒｒｅｓｔｅｒ，等引言１９８３年１月，在昆士兰州的中部埃默拉尔德，合成除虫菊酯类杀虫剂对大田棉铃虫失去了有效的控制。在此之前（Ｒｉｌｅｙ，１９８９），在美国因这类药剂取代了具有抗性和环境污染的有机氯制...     

Characterization of Escherichia coli Isolates Incriminated in Colisepticaemia in Mink

Colisepticaemia is a major health and economic concern for the mink industry, yet little information is available about the Escherichia coli that cause this disease. In this study, 40 E. coli, isolated from mink clinically diagnosed with colisepticaemia that had been submitted to the North Dakota State University Veterinary Diagnostic Laboratory, were randomly selected for characterization. These isolates were serotyped and screened for resistance to 18 antimicrobials, possession of transmissible R plasmids, and the presence of several virulence traits or genes using bioassays or the polymerase chain reaction. Several serotypes were identified that have previously been associated with septicaemia in other animal species. The majority of the isolates exhibited multiple antimicrobial resistance phenotypes. Common resistance phenotypes observed included those to tetracycline, sulfamethoxazole, streptomycin, ampicillin and kanamycin. Several of the isolates that could be studied by conjugation contained transmissible R plasmids coding for multiple antimicrobial resistance phenotypes. About half of the isolates produced colicin; all produced enterobactin; and all but one-quarter produced aerobactin. None of the isolates tested produced enterohaemolysin, and one-fifth were considered to be beta haemolytic. About half appeared to contain the gene encoding cytotoxic necrotizing factor-I; three contained the gene encoding EAE, but none appeared to contain the genes coding for LT, Sta/b, SLT-I/II or CNF-II toxins or K99 antigen. Approximately one-third of the isolates elaborated capsule. The results show that the E. coli isolates implicated in mink colisepticaemia possess similar virulence traits and antimicrobial resistance phenotypes to those associated with diarrhoeal diseases in food animals.     

Technical note: epimerization of ergopeptine alkaloids in organic and aqueous solvents

Purified ergopeptine alkaloids are often used in studies related to tall fescue toxicosis without regard to epimerization that occurs when ergopeptines are solvated. The objectives of this study were to measure the rates of alpha-ergocryptine epimerization to alpha-ergocryptinine at room temperature and at -40 degrees C, and to measure the rate of ergovaline epimerization to ergovalinine at 37 degrees C. Alpha-ergocryptine tartrate was stable (< 0.5% epimerization) in protic or aprotic solvents when stored at -40 degrees C for 20 to 52 d. At room temperature, alpha-ergocryptine epimerization in chloroform did not occur; epimerization was modest in acetone and acetonitrile (< 5%) but was substantial in methanol (78% by 38 d) and in a 70:30 water methanol mix (47% by 42 d). Ergovaline epimerization to ergovalinine occurred at 37 degrees C in 0.1 M phosphate buffers (pH 3, 7.5, and 9) in 9% aqueous solutions of fetal bovine serum (FBS), and in water, methanol, and acetonitrile. The degree of epimerization at 37 degrees C was solvent-dependent. Epimerization rates with respect to time were roughly linear in phosphate buffer (pH 3 only), water, methanol, and acetonitrile; epimerization rates resembled first-order kinetics in phosphate buffers (pH 7.5 and 9) and in the presence of FBS (pH 3, 7.5 and in Dulbecco's culture media). Epimerization equilibria (48 to 63% ergovaline) were reached within approximately 1 to 19 h. Results from this study indicate that researchers conducting studies with purified ergopeptines should carefully control the storage conditions of solvated ergopeptines and measure isomeric composition under the actual experimental conditions used in experiments.     

Effects of clenbuterol on body stores of polychlorinated dibenzofurans (PCDF) and dibenzo-p-dioxins (PCDD) in rats

Polychlorinated dibenzo-p-dioxins (PCDD) and polychlorinated dibenzofurans (PCDF), persistent pollutants that accumulate in the food chain, pose a risk to humans through consumption of tainted livestock. Clenbuterol, a leanness-enhancing agent, was tested for usefulness in PCDD/F body store reduction through body fat reduction (the predominant site of accumulation). To mimic the situation of contaminated animals, rats were given feed with or without a mixture of PCDD/F (0.6 to 2.7 ng/congener per day) for 10 d, followed by 16 d of feed with or without dietary clenbuterol (2 mg/kg feed). Clenbuterol reduced body fat by 28% (P < 0.05), increased muscle mass by 25% (P < 0.02), and decreased liver mass by 7% (P < 0.02). Although the concentrations of most PCDD/F per gram of fat were slightly increased after clenbuterol treatment, the total amount of PCDD/F that remained in fat was reduced by approximately 30%. Muscle PCDD/F concentrations and total burden were decreased by clenbuterol. In contrast, clenbuterol tended to increase concentration, but not total burden of PCDD/F in livers. One congener known to be rapidly metabolized and excreted, 2,3,7,8-TCDF, was the exception to this increase, decreasing 40% with clenbuterol treatment. This was also the congener that showed the greatest reduction in both fat and muscle. Examination of the ratio of PCDD/F in liver and fat revealed that clenbuterol increased the liver's share of the body burden of PCDD/F, from 38 to 75%. In a remediation/disposal context, these findings would be beneficial if clenbuterol lowered the meat and carcass burden of PCDD/F to safe levels, requiring only livers to be disposed of as hazardous waste.