Inhibition of multidrug-resistant Staphylococci by sodium chlorate and select nitro- and medium chain fatty acid compounds.

AIMS: Determine the antimicrobial effects of 5 µmol ml-1 sodium chlorate, 9 µmol ml-1 nitroethane or 2-nitropropanol as well as lauric acid, myristic acid and the glycerol ester of lauric acid Lauricidin® , each at 5 mg ml-1 , against representative methicillin-resistant staphylococci, important mastitis- and opportunistic dermal-pathogens of humans and livestock. METHODS AND RESULTS: Three methicillin-resistant Staphylococcus aureus and two methicillin-resistant coagulase-negative staphylococci were cultured at 39°C in 5 µmol ml-1 nitrate-supplemented half-strength Brain Heart Infusion broth treated without or with the potential inhibitors. Results revealed that 2-nitropropanol was the most potent and persistent of all compounds tested, achieving 58-99% decreases in mean specific growth rates and maximum optical densities when compared with untreated controls. Growth inhibition did not persist by cultures treated solely with chlorate or nitroethane, with adaptation occurring by different mechanisms after 7 h. Adaptation did not occur in cultures co-treated with nitroethane and chlorate. The medium chain fatty acid compounds had modest effects on all the staphylococci tested except the coagulase-negative Staphylococcus epidermidis strain NKR1. CONCLUSIONS: The antimicrobial activity of nitrocompounds, chlorate and medium chain fatty acid compounds against different methicillin-resistant staphylococci varied in potency. SIGNIFICANCE AND IMPACT OF THE STUDY: Results suggest that differential antimicrobial activities exhibited by mechanistically dissimilar inhibitors against methicillin-resistant staphylococci may yield potential opportunities to combine the treatments to overcome their individual limitations and broaden their activity against other mastitis and dermal pathogens.

The Role of Direct-Fed Microbials in Conventional Livestock Production.

Supplementation of direct-fed microbials (DFM) as a means to improve the health and performance of livestock has generated significant interest over the past 15+ years. A driving force for this increased interest in DFM is to reduce or eliminate the use of low-dose antibiotics in livestock production. This increased attention toward DFM supplementation has generated an extensive body of research. This effort has resulted in conflicting reports. Although there has been considerable variation in the design of these studies, one of the main causes for this lack of consistency may be attributed to the variation in the experimental immune challenge incorporated to evaluate DFM supplementation. Taking into account the experimental immune challenge, there is strong evidence to suggest that DFM supplementation may have an impact on the immune response, overall health, and performance of livestock.

Interactions between oil substrates and glucose on pure cultures of ruminal lipase-producing bacteria.

The hydrolysis of free fatty acids from lipids is a prerequisite for biohydrogenation, a process that effectively saturates free fatty acids. Anaerovibrio lipolyticus 5s and Butyrivibrio fibrisolvens have long been thought to be the major contributors to ruminal lipolysis; however, Propionibacterium avidum and acnes recently have been identified as contributing lipase activity in the rumen. In order to further characterize the lipase activity of these bacterial populations, each was grown with three different lipid substrates, olive oil, corn oil, and flaxseed oil (3 %). Because different finishing rations contain varying levels of glycogen (a source of free glucose) this study also documented the effects of glucose on lipolysis. P. avidum and A. lipolyticus 5s demonstrated the most rapid rates (P < 0.05) of lipolysis for cultures grown with olive oil and flaxseed oil, respectively. A. lipolyticus, B. fibrisolvens, and P. avidum more effectively hydrolyzed flaxseed oil than olive oil or corn oil, especially in the presence of 0.02 % glucose. Conversely, P. acnes hydrolyzed corn oil more readily than olive oil or flaxseed oil and glucose had no effect on lipolytic rate. Thus, these bacterial species demonstrated different specificities for oil substrates and different sensitivities to glucose.

Competitive effect of commensal faecal bacteria from growing swine fed chlortetracycline-supplemented feed on ß-haemolytic Escherichia coli strains with multiple antimicrobial resistance plasmids.

AIMS: To determine the differences in competitive fitness among Escherichia coli strains with different plasmid profiles when grown in suspension with commensal faecal bacteria from growing swine fed chlortetracycline-supplemented or unsupplemented diets. METHODS AND RESULTS: Five multiple drug-resistant (MDR) E. coli strains that possessed 0, 2, 6 or 8 plasmids were inoculated into anoxic faecal cultures from swine fed an unsupplemented (control) or chlortetracycline (50 g ton(-1))-supplemented (experimental) diet. On days 21 of chlortetracycline supplementation, faecal growth competition studies were performed. MDR E. coli were enumerated at 0, 6 and 24 h. The plasmid-free strain was below culturable limits in both the control and experimental cultures by 24 h. For each plasmid-bearing strain, there was no statistically significant difference in population CFU ml(-1) (P < 0.05) between the control and experimental cultures. CONCLUSIONS: There was no significant effect on the faecal microflora, owing to the inclusion of chlortetracycline, in the swine diets, that affected the growth of E. coli in the competition studies employed. Furthermore, these results suggest that the cost of maintaining plasmids in these E. coli strains had little influence on survivability. SIGNIFICANCE AND IMPACT OF STUDY: Mutations that led to antimicrobial resistance may have a greater impact on survivability than multiple plasmid carriage.

Probiotics, prebiotics and competitive exclusion for prophylaxis against bacterial disease.

The microbial population of the intestinal tract is a complex natural resource that can be utilized in an effort to reduce the impact of pathogenic bacteria that affect animal production and efficiency, as well as the safety of food products. Strategies have been devised to reduce the populations of food-borne pathogenic bacteria in animals at the on-farm stage. Many of these techniques rely on harnessing the natural competitive nature of bacteria to eliminate pathogens that negatively impact animal production or food safety. Thus feed products that are classified as probiotics, prebiotics and competitive exclusion cultures have been utilized as pathogen reduction strategies in food animals with varying degrees of success. The efficacy of these products is often due to specific microbial ecological factors that alter the competitive pressures experienced by the microbial population of the gut. A few products have been shown to be effective under field conditions and many have shown indications of effectiveness under experimental conditions and as a result probiotic products are widely used in all animal species and nearly all production systems. This review explores the ecology behind the efficacy of these products against pathogens found in food animals, including those that enter the food chain and impact human consumers.

The effect of an experimental chlorate product on Salmonella recovery of turkeys when administered prior to feed and water withdrawal.

Previously, an experimental chlorate product (ECP) has been observed to reduce Escherichia coli and Salmonella infections in swine, cattle, and broilers. The following studies were performed to investigate the effects of different concentrations and durations of administering ECP on crop and ceca Salmonella typhimurium (ST) colonization of turkeys. In 2 separate trials, each conducted with 2 replicates, 15-wk-old turkey toms were challenged with 10(7) to 10(9) cfu of ST. In Experiment 1, toms were administered 0, 0.5, 1.0, 2.0, or 4.0x of ECP (a 1.0x concentration is equivalent to a 15 mM chlorate ion concentration) in the drinking water for 38 h. In Experiment 2, toms were administered a 2x concentration of ECP in the drinking water for 0, 14, 26, or 38 h prior to water withdrawal. All treatments were followed by a 10-h water withdrawal and an 8-h feed withdrawal prior to organ sampling. In Experiment 1, turkeys provided ECP had significantly (P < 0.05) lower populations and incidences of crop (>1.4 log reduction) and ceca (>0.6 log reduction) ST as compared with control birds (2.1 and 0.94 log ST average for all trials, respectively), with little or no additional benefit from administration of higher ECP concentrations. In Experiment 2, toms provided ECP had lower populations of crop (>2.2 log reduction) and ceca (>1.5 log reduction) ST when compared with controls (3.1 and 1.8 log ST, respectively). Again, there appeared to be little benefit in longer administration intervals on quantitative reduction of ST. These experiments suggest that the ECP significantly reduces Salmonella colonization in commercial turkeys when administered prior to feed and water withdrawal.

Effect of ionophore supplementation on the incidence of Escherichia coli O157:H7 and Salmonella and antimicrobial susceptibility of fecal coliforms in Stocker cattle.

To examine the effect of ionophore supplementation on fecal shedding of Escherichia coli O157:H7 and Salmonella, crossbred beef calves (n=113; mean body weight [BW], 243 kg) were fed a mineral supplement with ionophore (1.76 g lasalocid/kg) for 61 days (d). Control calves received an identical mineral supplement without lasalocid. Calves were pastured on fescue/bermudagrass paddocks and supplemented with a corn/wheat midds/soybean meal supplement (1.5% of BW/d). Upon arrival, cattle were fed a commercial receiving ration containing 1 g chlorotetracycline/kg for 10 d. Sick calves were administered one or a combination of the following: Nuflor (florfenicol), Baytril (eurofloxacin), Micotil (tilmicosin), or LA 200 (oxytetracycline). Fecal samples were collected immediately prior to ionophore supplementation, approximately midway and at the end of the experimental period (60 d total ionophore feeding) for isolation of E. coli O157:H7 and Salmonella. Putative fecal coliforms were also isolated at these sampling times and examined for antimicrobial susceptibility. The study was replicated over a two year period (year 1, n=53 head; year 2, n=60 head). Ionophore supplementation had no effect (p>0.10) on the incidence of calves shedding E. coli O157:H7 or Salmonella. The percentage of calves shedding E. coli O157:H7 varied throughout the experimental period from 0 to 30%, while Salmonella was cultured from only three calves over the 2-year experimental period. Antimicrobial susceptibility profiles of putative fecal coliforms were consistent with antibiotic treatments administered during the study (observed resistance to chlortetracycline, florfenicol, oxytetracycline), while only one treatment effect was observed. Ionophore treatment resulted in a significantly higher number of coliform isolates resistant to ampicillin compared to controls in year 1, but not year 2. A number of fecal coliform isolates demonstrated resistance to multiple antibiotics, however, this was not affected (p>0.10) by ionophore supplementation. Mineral intakes, BW gain, and the number of sick calves were similar (p>0.10) among treatments. Ionophore supplementation had no affect on fecal shedding of E. coli O157:H7 or Salmonella and a negligible impact on antimicrobial susceptibility patterns of fecal coliforms in beef calves.

Effect of ractopamine HCl supplementation on fecal shedding of Escherichia coli O157:H7 and Salmonella in feedlot cattle.

The effects of the beta-agonist ractopamine, recently approved for use in feedlot cattle to improve carcass quality and performance, on fecal shedding Escherichia coli O157:H7 and Salmonella in feedlot cattle was examined. In the first study, 20 feedlot steers and heifers were randomly assigned to receive ractopamine or no ractopamine (control) by way of oral bolus for 28 days. Fecal samples were collected daily, and shedding of E. coli O157:H7 determined. When examined during the entire 28-day experimental period, ractopamine decreased (P = 0.0006) the percentage of cattle shedding E. coli O157:H7 (58% vs. 42% for control and ractopamine treatments, respectively). A second study was conducted in a commercial feedlot facility in the southwestern United States. Eighteen pens of cross-bred beef heifers (approximately 100 head/pen and 9 pens/treatment) were randomly assigned to receive either 0 (control) or 200 mg ractopamine/head x d(-1). Fresh fecal samples (30/pen) were collected off the pen floor before ractopamine supplementation and again after approximately 28 days of ractopamine supplementation (within a few days of slaughter); the samples were cultured for E. coli O157:H7 and Salmonella. The percentage of animals shedding E. coli O157:H7 was decreased when data were pooled across replicates (P = 0.05) in ractopamine-treated cattle compared with controls. The percentage of animals shedding Salmonella tended to be higher (P = 0.08) with the ractopamine treatment when data were pooled across replicates. Although further research is required to confirm these results, the potential food safety implications of this research are intriguing.

Effects of alfalfa and fructooligosaccharide on molting parameters and bone qualities using dual energy X-ray absorptiometry and conventional bone assays.

The aim of this study was to evaluate effects of alfalfa and fructooligosaccharides (FOS) on molting performance and bone parameters compared with the conventional feed withdrawal molting procedure. A total of 36 Single Comb White Leghorn hens (84 wk of age) were used for this experiment. The hens were divided into 6 treatment groups with 6 birds per treatment: pre-trial control (PC), full fed (FF), feed withdrawal (FW), 100% alfalfa (A100), A100 + 0.375% FOS (A100L), and A100 + 0.75% FOS (A100H). At the end of the 9-d molt period, hens were euthanized, and tibia and femurs were collected to evaluate bone qualities using dual-energy x-ray absorptiometry (DXA), Instron (Model 1011 Instron Universal Testing Machine, Instron Corp., Canton. MA), and conventional bone assays. Egg production was recorded during the molting period to evaluate first day out of production, and ovary was also collected to measure ovary weight. Alfalfa molting diets had comparable molting parameters, such as percentage of BW loss, ovary weight, and first day out of egg production, to the conventional feed withdrawal molting procedure, and FOS supplementation did not have any detrimental effects on molting performance. Conventional bone assay and DXA results suggest that hens lose a considerable amount of bone minerals during a molting period. The tibia and femur bone strengths of the FF, FW, A100, and A100L hens were significantly lower than the PC hens, whereas hens fed A100H had similar tibia bone breaking strength to that of the PC hens. The bone parameters measured by conventional assays, bone breaking strength measured by Instron, and bone density and mineral content measured by DXA were highly correlated to each other.

Comparison of spontaneous antibiotic resistance frequency of Salmonella Typhimurium growth in glucose amended continuous culture at slow and fast dilution rates.

The objective of the study was to determine the frequency of spontaneous acquisition of resistance to select antibiotics by Salmonella Typhimurium (ST) when grown in glucose amended continuous flow culture at slow (D = 0.025 h(-1)) or fast (D = 0.27 h(-1)) dilution rates. The bacterium was grown in LB minimal medium (pH 6.25) containing no antibiotics. Upon achieving steady state, samples were plated to tryptic soy agar (TSA) alone or supplemented (per ml) with 2 and 16 microg oxytetracycline, 4 and 16 microg tetracycline, 2 and 64 microg kanamycin, and 0.25 and 2 microg enrofloxacin. Regardless of growth rate, CFU of resistant ST from the TSA containing antibiotics was less than 2 x 10(1) except for 2 microg kanamycin and 0.25 microg enrofloxacin treatments (higher than 1 x 10(9) and 4 x 10(7) CFU of resistant ST for trials 1 and 2, respectively). Frequency of recovering resistant ST from the TSA containing the higher antibiotic concentrations was less than 1 in 10(9) for all antibiotics, but was higher on the media containing 2 microg kanamycin and 0.25 microg enrofloxacin at both slow and fast growth rates. In general, minimal susceptibility differences were detected for isolates from slow and fast dilution rates.

Effects of drinking water treatment on susceptibility of laying hens to Salmonella enteritidis during forced molt.

Feed deprivation is used in the layer industry to induce molting and stimulate multiple egg-laying cycles in laying hens. Unfortunately, the stress involved increases susceptibility to Salmonella enteritidis (SE), the risk of SE-positive eggs, and incidence of SE in internal organs. Leghorn hens over 50 wk of age were divided into 4 treatment groups of 12 hens each in experiment 1 and 3 treatment groups of 12 hens in experiments 2 and 3; hens were placed in individual laying hen cages. Treatment groups were 1) nonmolted (NM) and received feed and distilled water for 9 d, 2) force molted by feed removal for 9 d and received distilled water, 3) force molted by feed removal for 9 d and received 0.5% lactic acid (LA) in distilled water. An additional group (4) in experiment 1 only was force molted by feed removal for 9 d and received 0.5% acetic acid in distilled water. Seven days before feed removal hens were exposed to an 8L:16D photoperiod, which was continued throughout the experiment. Individual hens among all treatments were challenged orally with 10(4) SE on d 4 of feed removal. When compared with the NM treatments, weight losses were significantly higher in the M treatments, regardless of water treatments. When compared with NM treatments, crop pH was significantly higher in the M treatment receiving distilled water. Crop pH was reduced to that of the NM controls by 0.5% acetic acid in the drinking water. No consistent significant changes were observed for volatile fatty acids. The number of hens positive for SE in crop and ceca after culture and the number of SE per crop and per gram of cecal contents were higher in the M treatments, when compared with the NM treatments, but there was no effect of addition of either of the acids to the drinking water. Additional research using different acid treatment regimens may provide a tool for reducing the incidence of SE in eggs and internal organs during and following molting of laying hens.

Alfalfa as a single dietary source for molt induction in laying hens.

Molting is a process by which a hen's reproductive tract is rejuvenated prior to the beginning of a laying cycle. This process is often artificially induced in commercial settings in order to extend the productive life of a flock of hens. The most common method for the induction of molt is feed withdrawal for a period of several days. It has been noted that feed withdrawal, while effective in inducing molt and allowing an adequate reproductive rest period for the hen, may cause deleterious effects on the animal. This has prompted the investigation of alternatives to feed deprivation for the induction of molt in commercial laying hens. This study involved feeding alfalfa to hens to assess its ability to induce molt. Results show that alfalfa meal and alfalfa pelleted diets were equally effective as feed withdrawal in causing ovary weight regression in birds. Molted hens induced by alfalfa diets exhibited postmolt levels of egg production over a twelve week period that were similar to that of hens molted by feed withdrawal. The postmolt eggs laid by hens molted by alfalfa were of comparable quality to eggs from feed deprived hens. Alfalfa, a fibrous feed with low metabolizable energy, may be provided to hens on an ad libitum basis for an effective molt induction that retains comparable egg quality and production.

Review on the role of dietary zinc in poultry nutrition, immunity, and reproduction.

Zinc is an important nutrient in animal metabolism. In poultry, zinc serves not only as a nutrient but can also be used as a dietary supplement to manipulate the reproductive system of the bird. This article summarizes the general biochemistry, physiology, and nutritional aspects of zinc metabolism to provide a brief overview on what is known regarding zinc. The potential role of zinc in poultry immune response, Salmonella infection, and molting are emphasized.

Using a feed-grade zinc propionate to achieve molt induction in laying hens and retain postmolt egg production and quality.

A commercial-feed-grade form of zinc propionate was examined as a potential feed amendment at a concentration of 1% zinc to induce molt in 90-wk-old hens. Dietary treatments consisted of 4 treatment groups of 28 birds each randomly assigned to either (1) molted conventionally by feed withdrawal, (2) 1% zinc as Zn acetate, (3) 1% zinc as Zn propionate, or (4) nonmolted control for 9 d. Ovary weights of hens fed Zn acetate or Zn propionate were not significantly different from each other, but hens fed Zn acetate or Zn propionate were significantly (p<0.05) lighter than the ovary weight of nonmolted control hens. Zinc concentrations in the kidney and liver were significantly (p<0.05) increased in both Zn acetate- and Zn propionate-molted hens when compared to either nonmolted control-fed hens or feed-withdrawal molted hens. Over the entire 3-mo postmolt period, there were no significant differences in interior or exterior egg qualities among the four treatments. Egg production of hens fed Zn acetate was significantly lower than feed-withdrawal hens, Zn propionate-fed hens, or nonmolted control hens (p<0.05). The data of the current study demonstrated that feeding a feed grade of Zn propionate (1% Zn)-supplemented diet can induce molt and retain postmolt egg quality and production comparable to hens molted by feed withdrawal.

Recent pre-harvest supplementation strategies to reduce carriage and shedding of zoonotic enteric bacterial pathogens in food animals.

Food-borne bacterial illnesses strike more than 76 million North Americans each year. Many of these illnesses are caused by animal-derived foodstuffs. Slaughter and processing plants do an outstanding job in reducing bacterial contamination after slaughter and during further processing, yet food-borne illnesses still occur at an unacceptable frequency. Thus, it is imperative to widen the window of action against pathogenic bacteria. Attacking pathogens on the farm or in the feedlot will improve food safety all the way to the consumer's fork. Because of the potential improvement in overall food safety that pre-harvest intervention strategies can provide, a broad range of preslaughter intervention strategies are currently under investigation. Potential interventions include direct anti-pathogen strategies, competitive enhancement strategies and animal management strategies. Included in these strategies are competitive exclusion, probiotics, prebiotics, antibiotics, antibacterial proteins, vaccination, bacteriophage, diet, and water trough interventions. The parallel and simultaneous application of one or more preslaughter strategies has the potential to synergistically reduce the incidence of human food-borne illnesses by erecting multiple hurdles, thus preventing entry of pathogens into the food chain. This review emphasizes work with Escherichia coli O157:H7 to illustrate the various strategies.

Effects of high zinc diets using zinc propionate on molt induction, organs, and postmolt egg production and quality in laying hens.

This study was conducted to determine the ability of an alternative salt form of 1% Zn, Zn propionate, to induce molt in 66-wk-old hens. The hens were randomly assigned to 4 treatment groups of 27 or 28 birds each: a) molted conventionally by feed withdrawal, b) 1% Zn as Zn acetate, c) 1% Zn as Zn propionate, or d) nonmolted control for 9 d. Feed intake was (P < 0.05) depressed in Zn acetate and Zn propionate hens when compared with nonmolted control hens during the 9 d. Ovary weights of hens undergoing feed withdrawal, Zn acetate, or Zn propionate were not (P > 0.05) different from each other, but all were (P < 0.05) lighter than the ovary weights of nonmolted control hens. Zinc concentrations in the kidney and liver were (P < 0.05) increased in Zn acetate and Zn propionate molted hens when compared with nonmolted hens on the control diet or hens molted by feed withdrawal. Bone ash values were (P < 0.05) increased for Zn acetate and Zn propionate molted hens or nonmolted control hens as compared with molted hens on feed withdrawal. Over the entire 3-mo postmolt period, there were no significant differences in interior egg qualities, but egg weights from hens fed Zn propionate were (P < 0.05) heavier than those from hens on feed withdrawal. The data of the current study demonstrated that feeding a Zn propionate (1% zinc)-supplemented diet can induce molt.

Drought associated poisoning of cattle in South Texas by the high quality forage legume Leucaena leucocephala.

Approximately 80 head of yearling cattle grazing on 680 acres exhibited signs of Leucaena leucocephala toxicosis, which was confirmed in 3 animals by detection of 3-hydroxy-4 (IH)-pyridone, the metabolite of the poisonous principle mimosine, in their urine. The animals had grazed leucaena almost exclusively due to lack of alternative forage resulting from drought conditions. Toxicosis from this otherwise high quality forage would likely not have occurred had animals consumed lower amounts of leucaena and could probably have been prevented, as it has been elsewhere, had the animals been colonized with Synergistes jonesii, a beneficial ruminal bacterium capable of degrading the toxic metabolites.

Short-chain fatty acids affect cell-association and invasion of HEp-2 cells by Salmonella typhimurium.

This study demonstrates that the growth of S. typhimurium in Luria Bertani broth supplemented with acetate, propionate, butyrate, or a mixture of the three SCFA, affected cell-association and the ability to invade cultured HEp-2 cells. Cell-association and invasion was determined after growth for 4 h of growth in the presence of the SCFA at pH 6 and 7. The results suggest that the growth rate of the culture may have affected cell-association and invasion since accompanying the significant decrease in growth rate in the presence of SCFA at pH 6 was a decrease in cell-association and invasion. However, the results also suggest that the individual SCFA may play a role in modulating cell-association and the invasion phenotype and the regulation of cell-association and invasion by the SCFA was dependent on the concentration and the pH of the medium. Although the growth rates were similar for S. typhimurium in the SCFA mixture, butyrate (100 mM) and propionate (50 mM) at pH 6, differences in cell-association and invasion were observed among these cultures. Also, at pH 7, differences were observed among the SCFA treatments even though the growth rates were similar.