Sara A. Cutler

Dietary Inclusion of Colicin E1 Is Effective in Preventing Postweaning Diarrhea Caused by F18-Positive Escherichia coli in Pigs

ABSTRACT With worldwide concern over the use of antibiotics in animal agriculture and their contribution to the spread of antibiotic resistance, alternatives to conventional antibiotics are needed. Previous research in our laboratories has shown that colicin E1 is effective against some Escherichia coli strains responsible for postweaning diarrhea (PWD) in vitro. In this study we examined the efficacy of the dietary inclusion of colicin E1 in preventing experimentally induced PWD caused by F18-positive enterotoxigenic E. coli in young pigs. Twenty-four weaned pigs (23 days of age), identified by genotyping to be susceptible to F18-positive E. coli infections, were individually housed and fed diets containing 0, 11, or 16.5 mg colicin E1/kg diet. Two days after the start of the trial, all animals were orally inoculated with 1 × 109 CFU of each of two F18-positive E. coli strains isolated from pigs with PWD. The dietary inclusion of colicin E1 decreased the incidence and severity of PWD caused by F18-positive enterotoxigenic E. coli and improved the growth performance of the piglets. Additionally, the reduced incidence of PWD due to dietary colicin E1, lowered the levels of expression of the genes for interleukin 1β and tumor necrosis factor beta in ileal tissues from these animals. The dietary inclusion of colicin E1 may be an effective alternative to conventional antibiotics in the diets of weaning pigs for the prevention of PWD caused by F18-positive enterotoxigenic E. coli.

Response to dietary phosphorus deficiency is affected by genetic background in growing pigs

Concern over the environmental effect of P excretion from pig production has led to reduced dietary P supplementation. To examine how genetics influence P utilization, 94 gilts sired by 2 genetic lines (PIC337 and PIC280) were housed individually and fed either a P-adequate diet (PA) or a 20% P-deficient diet (PD) for 14 wk. Initially and monthly, blood samples were collected and BW recorded after an overnight fast. Growth performance and plasma indicators of P status were determined monthly. At the end of the trial, carcass traits, meat quality, bone strength, and ash percentage were determined. Pigs fed the PD diet had decreased (P < 0.05) plasma P concentrations and poorer G:F (P < 0.05) over the length of the trial. After 4 wk on trial, pigs fed the PD diet had increased (P < 0.05) plasma 1,25(OH)(2)D(3) and decreased (P < 0.05) plasma parathyroid hormone compared with those fed the PA diet. At the end of the trial, pigs fed the PD diet had decreased (P < 0.05) BW, HCW, and percentage fat-free lean and tended to have decreased LM area (P = 0.06) and marbling (P = 0.09) and greater (P = 0.12) 10th-rib backfat than pigs fed the PA diet. Additionally, animals fed the PD diet had weaker bones and also decreased (P < 0.05) ash percentage and increased (P < 0.05) concentrations of 1alpha-hydroxylase and parathyroid hormone receptor mRNA in kidney tissue. Regardless of dietary treatment, PIC337-sired pigs consumed more feed and gained more BW than their PIC280-sired counterparts (P < 0.05) during the study. The PIC337-sired pigs also had greater (P < 0.05) HCW, larger (P < 0.01) LM area, and tended to have (P = 0.07) greater dressing percentage. Meat from the PIC337-sired pigs also tended to have greater (P = 0.12) concentrations of lactate but decreased (P = 0.07) concentrations of total glucose units 24 h postslaughter. Although plasma 1,25(OH)(2)D(3) concentrations were elevated (P < 0.05) in all the animals fed the PD diet, this elevation due to P deficiency tended (P = 0.09) to be greater in the PIC337-sired pigs after 12 wk on the treatment. The PIC337-sired pigs had stronger (P < 0.01) bones with greater ash percentage than the PIC280-sired pigs. The difference in the strength of the radii between the PIC337-sired pigs fed the PA and PD diets was greater than their PIC280-sired counterparts, which resulted in sire line x treatment interactions (P < 0.05). These data indicate differing mechanisms of P utilization between these genetic lines. Elucidating these mechanisms may lead to strategies to increase efficiency of growth in a more environmentally friendly manner.

Inhibitory activity of colicin E1 against Listeria monocytogenes.

Colicins are gram-negative bacteriocins produced by and effective against Escherichia coli and related species. Colicin E1 (ColE1) is composed of three functional domains, which collectively have a pore-forming effect on targeted bacteria. ColE1 binding and translocation domains are highly specific in contrast to the pore-forming domain, implying that ColE1 could be broadly effective. In this study, the activity of ColE1 against Listeria monocytogenes was evaluated in broth and on surfaces of ready-to-eat products. Individual strains of L. monocytogenes were examined in broth containing ColE1 at 0, 0.1, 1, or 10 microg/ml. Although strain differences in sensitivity to ColE1 existed, growth was significantly reduced in all strains at doses as low as 0.1 microg/ml. Sterilized ham slices were submerged in a five-strain L. monocytogenes cocktail (either 7 or 4 log CFU/ ml) and placed in vacuum packages containing 0, 1, 5, 10, 25, or 50 microg of ColE1. Ham slices were then stored at 4 or 10 degrees C, and samples were removed and examined for L. monocytogenes after 1, 3, 7, and 14 days. Reduction of L. monocytogenes by ColE1 was dependent on initial inoculum concentration and storage temperature. For slices stored at 4 degrees C, treatment with 25 microg reduced Listeria growth below detection limits for the slices inoculated with 4 log CFU/ml for the entire 14 days, whereas for the 7-log CFU/ml slices, growth was detected at 7 days postinoculation. For slices stored at 10 degrees C, 10 microg/ml ColE1 significantly inhibited growth of L. monocytogenes for up to 3 days for both inoculation groups. These data indicate that ColE1 is highly effective against Listeria.

Effects of Lactobacilli and lactose on Salmonella typhimurium colonisation and microbial fermentation in the crop of the young turkey

1. Three experiments were performed to examine the effects of Lactobacilli and lactose on microbial fermentation and Salmonella enterica serovar Typhimurium colonisation in the crop of the young turkey. 2. The following carboxylic acids were detected in the crop ingesta: formic, acetic, butyric, lactic, valeric, caproic, oxalic, phenyl acetic, succinic and fumaric; propionic, isobutyric and isovaleric acids were not detectable. 3. At the beginning of the night, there were considerable quantities of ingesta in the crop of young turkeys. During the scotophase, there were progressive reductions in the contents and pH. Moreover, there were linear increases in the concentration of lactic, valeric and caproic acids (by approximately 7-fold over 8 h). Much smaller changes in crop pH were observed in the study where dietary treatments of Lactobacilli were not included. 4. Chronic addition of lactose or Lactobacilli to the diet exerted modest effects on the carboxylic acid concentration in the crop contents but did not consistently influence colonisation of the crop by Salmonella enterica serovar Typhimurium. 5. Young turkeys confine eating to the hours of illumination (photophase) with a peak in consumption prior to the subjective dusk.

Application of colicin E1 as a prefabrication intervention strategy.

Colicin E1 (ColE1) is a bacteriocin produced by and effective against Escherichia coli and related species. The current study examined ColE1 as a potential intervention strategy for controlling E. coli O157:H7 contamination on beef carcasses. Untrimmed beef round roasts were cut into sample sizes of 5.08 by 2.52 by 5.08 cm, with an adipose layer covering an entire surface of lean beef. Samples were placed on sterile metal hooks and inoculated with E. coli O157:H7 at a level of 5 log CFU/ml in sterile tryptic soy broth. After inoculum attachment, ColE1 in doses of 0, 100 microg, 500 microg, and 1 mg/ml of 10 mM Tris, pH 7.6, was sprayed on the samples for a period of 10 min. Samples were evaluated at 0 and 30 min, 1, 2, 3, 4, and 5 days post-spraying at 10 degrees C for E. coli O157:H7 inhibition. Treating samples with 500 microg and 1 mg of ColE1 effectively inhibited E. coli O157:H7 growth. When these doses were applied to samples inoculated with E. coli WS 3331, E. coli contamination was reduced by 4 and 7 log CFU/cm2, respectively, compared with the untreated control samples. In strain WS 3331, treatment with 1 mg ColE1 significantly inhibited growth of E. coli O157:H7 compared with the untreated control during the entire study. ColE1 provided powerful reduction of E. coli O157:H7 as a beef carcass spray intervention.

A calcitonin receptor (CALCR) single nucleotide polymorphism is associated with growth performance and bone integrity in response to dietary phosphorus deficiency

Although concerns over the environmental impact of excess P in the excreta from pig production and governmental regulations have driven research toward reducing dietary supplementation of P to swine diets for over a decade, recent dramatic increases in feed costs have further motivated researchers to identify means to further reduce dietary P supplementation. We have demonstrated that genetic background impacts P utilization in young pigs and have identified genetic polymorphisms in several target genes related to mineral utilization. In this study, we examined the impact of a SNP in the calcitonin receptor gene (CALCR) on P utilization in growing pigs. In Exp. 1, 36 gilts representing the 3 genotypes identified by this CALCR SNP (11, 12, and 22) were fed a P-adequate (PA) or a marginally P-deficient (approximately 20% less available P; PD) diet for 14 wk. As expected, P deficiency reduced plasma P concentration, bone strength, and mineral content (P < 0.05). However, the dietary P deficiency was mild enough to not affect the growth performance of these pigs. A genotype x dietary P interaction (P < 0.05) was observed in measures of bone integrity and mineral content, with the greatest reduction in bone strength and mineral content due to dietary P deficiency being associated with the allele 1. In Exp. 2, 168 pigs from a control line and low residual feed intake (RFI) line were genotyped for the CALCR SNP and fed a PA diet. As expected, pigs from the low RFI line consumed less feed but also gained less BW when compared with the control line (P < 0.05). Although ADFI did not differ between genotypes, pigs having the 11 genotype gained less BW (P < 0.05) than pigs having the 12 or 22 genotypes. Pigs of the 11 and 12 genotypes had bones that tolerated greater load when compared with animals having the 22 genotype (P < 0.05). A similar trend was observed in bone modulus and ash % (P < 0.10). These data are supportive of the association of this CALCR SNP with bone integrity and its response to dietary P restriction. Although the allele 1 is associated with greater bone integrity and mineral content during adequate P nutrition, it is also associated with the greatest loss in bone integrity and mineral content in response to dietary P restriction. Understanding the underlying genetic mechanisms that regulate P utilization may lead to novel strategies to produce more environmentally friendly pigs.

Effects of atrazine on sexual maturation in female Japanese quail induced by photostimulation or exogenous gonadotropin

The herbicide atrazine has gained recent attention for its reported effects on reproduction in amphibians. The present study examined the putative effects of atrazine during sexual maturation in the photostimulated female Japanese quail. Furthermore, the effects of atrazine on birds administered exogenous gonadotropin (pregnant mare serum gonadotropin [PMSG]) were investigated. Atrazine was administered up to 1,000 ppm in the diet to female quail undergoing photoperiodically induced sexual maturation. At high dietary concentrations, atrazine exhibits signs of overt toxicity with reductions in growth, feed intake, and liver weights, but these effects were dependent on the timing of treatment administration. Atrazine did not influence the weights of reproductive tissues (ovary and oviduct) or circulating concentrations of luteinizing hormone (LH). However, high concentrations of atrazine depressed circulating concentrations of estradiol. Treatment with atrazine for four weeks during sexual maturation inhibited growth but did not affect any other parameter assessed (feed intake, liver, ovary, or oviduct weights or the circulating concentrations of LH and estradiol). In birds receiving daily injections of PMSG, atrazine reduced growth, feed intake, and liver weights. However, PMSG-induced gonadal and oviduct growth was not affected by atrazine. The present results suggest that dietary atrazine exhibits limited reproductive toxicity in female quail during sexual maturation and only at concentrations above ecological relevance.

Effects of egg antibody to components of inflammatory activation (phospholipase a 2 and Toll like receptor 4) on the response of young turkeys to Salmonella typhimurium challenge

The effect of passive immunization against two components of the inflammatory response (Toll-like receptor 4 and phospholipase A2) was examined in turkey poults receiving a challenge with Salmonella enterica serovar typhimurium (ST). The number of ST in the crop ingesta were not influenced by prior oral administration of egg antibody against Toll-like receptor 4 and phospholipase A2. There was a marked effect of the passive immunization on the crop contents 48 hours following ST challenge. The crops of the ST challenged non-immune sera controls were distended and fluid filled while those of poults receiving egg antibody contained ingesta. The dry weights of the ingesta in the crop of the ST challenges non-immune egg immunoglobulin receiving birds were 510% those receiving egg antibody to Toll-like receptor 4 or phospholipase A2. It is concluded that fluid accumulation in the crop of the young turkey following ST challenge may be a good model for bacterial enteric diseases where there is profuse watery diarrhoea. Moreover, passive immunization with antibody to Toll-like receptor 4 and phospholipase A2 reduces the symptoms associated with salmonellosis and possibility other enteric diseases.