Twenty-one market-age swine slaughtered in the University of Kentuckyabattoir were inoculated with fecal slurry containing two strains of nalidixic acidresistant Salmonella typhimurium on the ham, belly, and jowl regions on eachside of the carcass. Trial 1 revealed that a 10 s hot water spray was just aseffective as the 20 s spray in removing S. typhimurium, as there was nodistinguishable difference in population. The shorter flame singe (10 s) was aseffective as the 20 s application and the two chlorine solutions (100, 200 ppm)had similar results. The 2% lactic acid spray reduced S. typhimurium populationssignificantly more than the 1% treatment. Trial 2 compared the four mostefficient levels of each intervention method. Efficacy of the intervention methodswas observed in the following order: Hot water (10 s) andgt; Chlorine (50 ppm) =Lactic acid (2%) andgt; Flame (10 s). The effect of carcass area was significantfollowing the post treatment hot water rinse. The jowl area was least accessibleby the high pressure water spray. However after the treatment applications, hotwater rinse, and 24 h chill (2??C) there was no significant difference betweentreated and untreated carcasses or between carcass areas.

Non-typhoidal Salmonella enterica serovars continue to be the leading cause of foodborne illnesses in United States. Chlorine, chlorine related, and quaternary compounds are generally used for disinfecting carcasses and equipment in processing industries. The current study was aimed at understanding the inactivation kinetics of four Salmonella serovars to chlorine, chlorine dioxide and cetylpyridinium chloride (CPC). The transcriptomic responses to oxidative stress was investigated in stationary and log phase cells of S. Typhimurium. The study was also aimed at understanding the effect of the chemicals on the expression of virulence genes associated with the Salmonella Pathogenecity Island 1 (SPI1). The possible induction of the viable but nonculturable (VBNC) state in Salmonella due to CPC was also investigated. The inactivation parameters for each serovar and the chemical were estimated based on the Hom's model, ln (N/N0) = -k C^n T^m and it appeared that while disinfectant contact time was significant, biocide concentration in the overall disinfection was insignificant. This was true especially for chlorine and CPC with subtle differences observed between the serovars. The inactivation efficacy was, however, dependent on both concentration and the exposure time for chlorine dioxide. The highest degree of inactivation was obtained with chlorine followed by chlorine dioxide and CPC. Transcriptomic responses of S. Typhimurium revealed significant downregulation of several metabolic processes such as tricarboxylic acid cycle, oxidative phosphorylation, and amino acid biosynthesis in both log and stationary phase cells. Several stress related genes such as usp, rpoS and ompR were upregulated in the stationary phase cells. Majority of the virulence genes associated with the SPI1 were found to be downregulated for all the treatments. While treatment with chlorine and CPC caused downregulation of all the virulence genes, treatment with chlorine dioxide caused significant upregulation of few (hilC, invC, sipA and sipB) genes associated with the SPI1. Finally, the induction of VBNC state was not concluded as a result of treatment with CPC. However, significant percentage of cells (45 percent) with intact membrane was established based on the BacLight assayTM.

This multi-phase project was designed (1) to evaluate existing post-harvest process controls and intervention strategies used to reduce Escherichia coli O157:H7, (2) to evaluate the impacts of cattle source and environmental factors on Salmonella prevalence in bovine lymph nodes, and (3) to evaluate sanitary conditions of feedyards in South Texas. The ultimate goal of this project was to identify and implement measures that reduce E. coli O157:H7 in beef harvest facilities, and Salmonella prevalence in feedyards. To evaluate process control of E. coli O157:H7 throughout the beef harvest process, samples were collected from harvest floor processing areas at two commercial beef slaughter establishments, and enumerated for aerobic plate counts, E. coli/coliform, and Enterobacteriaceae. To survey existing Salmonella prevalence, bovine lymph nodes (n = 307) were collected from beef carcasses at a commercial beef processing plant. Lymph nodes were extracted from cattle sourced from seven feedyards. Salmonella prevalence in lymph nodes was found to be 0% in cattle sourced from only one of the seven yards. Lymph nodes from cattle sourced from the other feedyards yielded positive samples, with varying prevalence. Of the remaining six feedyards, one feedyard yielded 88.2% prevalence of Salmonella in bovine lymph nodes, which was significantly higher than all other feedyards (42.9, 40.0, 40.0, 24.0, and 4.0%). The prevalence of Salmonella in the feedlot environment was compared among three feedyards; one yard had 65.0% environmental prevalence of Salmonella, which was statistically higher than the other feedyards surveyed. Of the two remaining yards, one had 0% prevalence of Salmonella in fecal and soil samples, which was also the feedyard with 0% prevalence of Salmonella in lymph nodes. Findings include (1) the significance of effective sanitary dressing procedures and intervention strategies in a beef harvest environment, (2) that there is clear feedyard-to-feedyard variation with relation to Salmonella prevalence in bovine lymph nodes, and (3) that differences in environmental factors existed among feedyards although the reasons remain unclear.

No description available.

7. Zusammenfassung Untersuchungen zur Bedeutung der Magen-Darm-Flora auf das Immunsystem von Hühnern Nasser Ali Abou Hashem Elsayed Instiut für Bakteriologie und Mykologie, Veterinärmedizinische Fakultät, Universität Leipzig Leipzig, Juli, 2002 (128 Seiten, 60 Abbildungen, 21 Tabellen und 227 Referenzen) Viele komplexe bakterielle Kontrollmechanismen der Darmmikroflora sind am Ausschluss pathogener Keime beteiligt. Dysregulationen im gastrointestinalen Ökosystem können den schützenden Effekt der indigenen Mikroflora schwächen, was die Kolonisation von pathogenen Keimen im Darmkanal begünstigt. Dysregulationen im gastrointestinalen Ökosystem treten bei Jungtieren hauptsächlich unter Stress, bei der Futterumstellung oder Verdauungsstörungen in Erscheinung. Alle diese Situationen kommen bei Jungvögeln vor. Dem zu Folge ist es nicht überraschend, dass Jungvögel empfindlicher für die Kolonisation von pathogenen Darmerregern wie Salmonellen sind. Das Verstehen der gastrointestinalen Mikroflora ist nicht nur für die Entwicklung von Probiotika sondern auch für das Entwickeln neuer Präbiotika wichtig. 1. Effekt von 0.5% Inulin im Trinkwasser und 1% Leinsamen im Futter auf die natürliche Besiedlung junger Küken mit S. Enteriditis, die Zugaben von Inulin im Trinkwasser (I-Gruppe) oder Inulin und Leinsamen (IL-Gruppe) an die jungen Küken führte zur Reduzierung der Nachweisrate von S. Enteritidis bei natürlich besiedelten Küken. 2. Effekte von 0.5% Inulin im Trinkwasser und 1% Leinsamen im Futter auf die intestinale bakterielle Besiedlung und den Immunstatus von SPF Hühnern, die bakteriologischen Untersuchungen offenbarten einen signifikanten Abfall der aeroben Gesamtbakterienzahl im Dünndarm und in Zäkum, aber nicht in Kropf und Rektum, insbesondere von der dritten Versuchswoche in den IL- und I-Gruppen gegenüber den Kontrolltieren. Die Zahl der gramnegativen Bakterien war in der zweiten Woche im Kropf und Dünndarm in den IL- und I-Gruppen deutlich rückläufig sowie in der dritten Woche im Zäkum und Rektum aber nicht in der C-Gruppe. Die Bdellovibrionenzahl stieg signifikant nur im Zäkum und im Rektum der IL- und I-Gruppen an, dem gegenüber konnten in der C-Gruppe keine Bdellovibrionen isoliert werden. Bedeutsame Unterschiede der Körpergewichte zwischen all den Gruppen zeigten sich nicht, wobei das signifikante Ansteigen die Quotient aus Bursa Fabricii/Körpergewicht in den IL- und I-Gruppen gegenüber der C-Gruppe auffielen. 3. Effekte von Inulin auf die Darmflora, den Endotoxin- und PC-BP-Blutspiegel und den Immunstatus von Broilern, nach der ersten Woche der Inulin-Verabreichung zeigten sich kaum Unterschiede in den Gesamtzahlen der aeroben Bakterien im Zäkum in der I-Gruppe (mit Inulin) und C-Gruppe (ohne Inulin). Hingegen stieg die Gesamtkeimzahl in der I-Gruppe gegenüber der C-Gruppe deutlich an. Gramnegative Bakterien fielen zahlenmäßig nach 2 Wochen deutlich bei der I-Gruppe ab. Die Bdellovibrionen steigen nach einer Woche in dieser I-Gruppe etwas an. Ab der zweiten Woche bis zum Ende des Versuchs (also nach 5 Wochen) stieg die Bdellovibrionenzahl in der I-Gruppe signifikant gegenüber der C-Gruppe. Dem gegenüber fiel die C. perfringens-Zahl in dieser I-Gruppe ab, welches in der ersten Versuchswoche von geringer Signifikanz war. In der dritten Woche fielen die zäkalen C. perfringens-Zahlen deutlich in der I-Gruppe gegenüber der C-Gruppe ab, wobei die Clostridien negativ zu den Bdellovibrionen im Zäkum korrelierten. Die Entwicklungen der Körpergewichte im Vergleich der beiden Gruppen zeigten insbesondere ab der 5.Woche signifikante Vorteile in der I-Gruppe. Die Quotienten aus Bursa Fabricii/Körpergewicht und Pankreas/Körpergewicht nahmen deutlich 2 Wochen nach der Inulin-Verabreichung bis zum Versuchsende zu. Die Endotoxin-Blutspiegel reduzierten sich in der I-Gruppe im Vergleich zu den Kontrolltieren besonders deutlich zum Ende des Experiments in der 6. Woche. Ebenso war eine Reduktion des PC-BP-Blutspiegels in der I-Gruppe in der zweiten Woche feststellbar in der 5.Woche. Die PC-BP-und Endotoxin-Blutspiegel korrelierten negativ zu den Körpergewichtzunahmen. Ebenso wirkten sich Zunahmen der aeroben Gesamtkeimzahl, gramnegativen- und Clostridien-Zahlen ungünstig auf die Körpergewichtszunahmen aus. Andererseits zeigt sich hierbei eine positive Korrelation zwischen der Bdellovibrionenzahl und der Körpergewichtzunahme. 4. In den Untersuchungen zur Dynamik der normalen Darmflora im Zäkum der jungen Broiler und erwachsenen Zuchthühner, fanden wir eine umgekehrte Beziehung zwischen dem zahlenmäßigen Vorkommen von Bdellovibrionen und C. perfringens. Die Anzahl der Bdellovibrionen korrelierte negativ zu den aeroben Gesamtkeimzahlen, den gramnegativen und C. perfringens-Bakterienzahlen sowie Endotoxin- und PC-BP (phosphoryl choline-binding protein)-Blutspiegeln. Andererseits korrelierten sie positiv zu den Quotienten aus Bursa Fabricii/Körpergewicht und Pankreas/ Körpergewicht. Die Endotoxin- und PC-BP-Blutspiegel korrelierten positiv mit dem Anstieg der Gesamtkeimzahlen. Das Pankreas/Körpergewicht-Verhältnis korrelierte positiv zum Verhältnis Bursa Fabricii/Körpergewicht. Die Resultate der vorliegenden Untersuchungen weisen darauf hin, dass Inulin für Hühner schützende Effekte offeriert: 1) Deutlich abnehmende intestinale Keimzahlen von C. perfringens. 2 ) Anstieg der Bdellovibrionenzahl. 3 ) Abfall des Endotoxin-Blutspiegels. 4 ) Abfall des PC-BP-Blutspiegels. 5 ) Verbesserung des Immunstatus der Vögel. Aus unseren Resultaten und vorhergehenden Studien können wir feststellen, dass Inulin- und Leinsamen als Funktionelle Futtermittel bei Geflügel eingestuft werden können, weil sie das Risiko einiger Erkrankungen verringern (z.B. durch Salmonellen). Inulin und Leinsamenl besitzen förderliche Effekte auf die intestinale Mikroflora und verbessern den Immunstatus der Tiere. Sie spielen eine wichtige Rolle in der Erhaltung der Homöostase in den ökologischen Systems des Magen-Darm-Traktes. Weiterführende Untersuchungen wären wünschenswert in Bezug auf die diätetischen Effekte des Inulins auf die anderen wichtigen Vertreter der Darmflora bei Hühnern wie Laktobazillen und Bifidobakterien. Auch die Kombination von Inulin mit Probiotika (Synbiotika) und deren Rolle zur verbesserten lokalen Immunität des Gastrointestinaltrakts und der kompetiven Exclusion pathogener Bakterien wie Salmonella-Serovare wären zu untersuchen. / 6. Summary Investigations on the significance of the gastrointestinal flora for the immune system of chickens Nasser Ali Abou Hashem Elsayed Institute of Bacteriology and Mycology, Faculty of Veterinary Medicine, University of Leipzig. Leipzig, July, 2002 (128 pages, 60 figures, 21 tables and 227 references) Many complex bacterial control mechanisms are involved in regulating the composition of the gut microflora and in excluding intestinal pathogens. Imbalances in the gastrointestinal ecosystem can weaken the protective effect of the indigenous microflora which gives enteric pathogen a better chance to colonize in the gut. Imbalances in the ecosystem mainly occur in the young animal, during periods of stress, changes in the periods of nutritive application and/ or digestive disorders. The young bird combines all these situations. It therefore not surprise that the young bird is prone to the colonization with enteric pathogens such as salmonellae. Understanding the gastrointestinal microflora is essential not only for the development of probiotics but also for developing new prebiotics. 1- Effect of inulin 0.5% via drinking water and feed supplement with 10% linseed on the natural colonization of young chicks with S. Enteritidis was examined; the addition of inulin to the drinking water (I-group) or inulin and linseed to the young chicks (IL-group) reduced the incidence of a naturally intestinal colonization with S. Enteritidis. 2- Effect of inulin 0.5 % via drinking water and food supplemented with 1% linseed on the intestinal bacterial population and the immune status of SPF chicken was investigated, the bacteriological examinations revealed a significant decrease in total aerobic bacterial count in the small intestine and caecum only but not in the crop and rectum of the IL-group and I-group respectively at the third week of the trial in comparison with control birds. Gram-negative bacterial counts also were significantly decreased in crop and small intestine of IL-group and I-group at the second week and in the caecum and rectum at the third week but not in the C-group. Bdellovibrio bacteriovorus counts were significantly increase only in the caecum and rectum of the IL-group and I-group respectively at the second week but can not be isolated from the C-group. There was no important difference in BW between all groups but there was a significant increase in BF/BW ratio in the I- group and in the IL-group than in C-group indicated increase immune capacity of IL-group and I-group. 3- The effect of inulin on the intestinal flora, endotoxin and PC-BP blood levels and on the immune status of broiler chicken was investigated, There was no significant difference in the total aerobic bacterial counts in the caecum of I-group (with inulin) and C-group (without inulin) after one week of inulin administration but at the second week the total bacterial count by I-group was significantly decreased than in C-group. Gram-negative bacterial counts were also significantly decreased after 2 weeks of inulin administration by I-group, whereas the Bdellovibrio counts were slightly increased in I-group after one week of inulin administration. At the second week and until the end of the experiment (5 weeks) the Bdellovibrio counts were significantly increased in I-group than in C-group. On the other hand, there was a decrease in the C. perfringens caecal counts of I-group, which were slightly significant at the first week of the experiment. At the third week there was a strongly significant decrease in C. perfringens caecal counts of the I-group in comparison with C-group. There was also a negative correlation between C. perfringens and Bdellovibrio caecal counts. There were no significant changes in BW between I-group (with inulin) and C-group (without inulin) during the experiment but at the 5th week, there was a significant increase in the BW of I-group, whereas there was a significant increase in BF/BW ratio in the I-group after 2 weeks of inulin administration until the end of the experiment. The endotoxin level in the blood of I-group was significantly reduced in comparison with control birds especially at the end of the experiment at 6th week. There was also a significant reduction in phosphoryl choline binding protein (PC-BP) at the second week and at the 5th week in the blood of the I-group in comparison with control birds. The PC-BP, endotoxin levels in the blood of birds were negatively related to the growth rates of BW. Also at increasing total aerobic, gram-negative and clostridial bacterial counts there were an adverse effect on the BW, whereas there was a positive correlation between Bdellovibrio count and growth rate of BW of birds. 4- When studying the dynamic of normal caecal flora of young (broilers) and adult (breeders) chicken we have found a conversely relationship between Bdellovibrio and C. perfringens counts. The Bdellovibrio count was negatively related to total aerobic, gram-negative bacterial counts and endotoxin blood level. On the other hand Bdellovibrio caecal counts were positively related with the BF/BW and P/BW ratios and to body weight, whereas the increase in the endotoxin and PC-BP blood levels had negative effect on these immune ratios. The endotoxin blood levels were also found positively related to phosphoryl choline-binding protein (PC-BP) levels and to increasing in the total aerobic and gram-negative bacterial counts. P/BW ratio was positively related with BF/BW ratio. Results of the present studies indicate that inulin offer protection to chickens by: 1- Markedly decreased intestinal colonization by C. perfringens counts. 2- Increase in Bdellovibrio bacteriovorus counts. 3- Decrease in the endotoxin level of blood. 4- Decrease in the PC-BP level of blood. 5- Enhancing the immune status of the bird. From our results and previous studies, we can conclude that inulin and linseed can be classified as functional diat in poultry, because they reduce the risk of some diseases (like Salmonella). Inulin and linseed have enhanced effects on the intestinal microflora and on the immune status of birds and play important role in the maintenance of homeostasis in the ecological system. Additional research is required in order to determine the effect of dietary inulin on the other important intestinal flora of chicken like Lactobacillus and bifidobacteria. Also the combination of inulin with probiotic (synobiotic) and their role in enhancing the local immunity of gastrointestinal tract and the competitive exclusion against pathogenic bacteria like Salmonella.

Despite their beneficial effects, concerns have been raised about the role of antimicrobial growth promoters (AGP) in the emergence of antibiotic resistant bacteria. This study evaluated the effects of approved AGP on the emergence of antibiotic resistance in commensal E. coli and foodborne pathogen Salmonella. A survey of antibiotic resistance levels in commercial broiler chicken farms in the Fraser Valley (B.C.) and an experimental feeding trial were conducted from May 2004 to February 2005 and May to November 2005, respectively. The latter examined the effects of ten AGP formulations (bambermycin, penicillin, salinomycin, bacitracin, combination of salinomycin and bacitracin, chlortetracycline, virginiamycin 11ppm, virginiamycin 22ppm, monensin and narasin) on bird performance as well. Multiple antibiotic resistant commensal E. coli and Salmonella carrying virulence genes were found at commercial broiler chicken farms and therefore may serve as reservoirs for these genes. There was no significant difference between feed formulations on the phenotypic or genotypic characteristics of the isolates, except for tetracycline resistance gene tet(B). In the experimental feeding trial, broiler chickens were fed a diet including or excluding AGP. Birds were sampled prior to and weekly during feeding of the control and the AGPP containing diets. Although not detected on day 0, E. coli increased after day 7 to more than 9.9 log10 CFU/g in ceca. Multi-drug resistant E. coli were isolated from birds fed the ten AGP containing diets as well as the control diet. Except for penicillin, none of the AGP containing diets significantly improved bird performance compared to the control diet (P>0.05). Good management practices can significantly improve broiler chickens performance and decrease the mortality rate.

Foodborne diseases caused by non-typhoidal Salmonella represent an important public health problem worldwide (Zhao et al., 2003). The transmission of Salmonella between animals and humans has been well established in epidemiological studies. In the case of complicated illness caused by Salmonella where antibiotics need to be administered, treatment can be compromised if the infecting organism is resistant to the prescribed antimicrobial agent. This study and earlier studies have shown that many Salmonella carry temperate bacteriophages as lysogens. Many of these bacteriophages are capable of mediating generalised transduction (Schicklmaier and Schmieger, 1995; Schicklmaier et al., 1998; Mmolawa et al., 2002). Schmieger and Schicklmaier (1999) demonstrated that bacteriophages ES18 and PDT17 are capable of transduction of antibiotic resistance genes from DT104. Phage-mediated transduction of antibiotic resistance genes has been largely neglected in the study of genetic transfer of antibiotic resistance in bacteria. This study investigates whether bacteriophages exist in antibiotic resistant Salmonella isolates. Such temperate phages in antibiotic resistant isolates could play a significant role in the transfer of resistance to other species of enteric bacteria, such as E. coli. Molecular epidemiology studies of antibiotic resistance genes were undertaken with Salmonella isolates from chicken, pig and human sources that were subjected to PCR for ampicillin (blaTEM-1), tetracycline (tetA, tetB) and streptomycin (aadA1, aadA2, strA, strB) resistance genes as well as Class 1 integrons. The blaTEM-1 gene was widely detected in isolates from pigs and chickens but rarely detected in human isolates. The tetB gene was more commonly found in pig isolates, while the tetA gene was associated with tetracycline resistance in chicken isolates. The strA and strB genes were responsible for streptomycin resistance in the S. Typhimurium isolates while the aadA1 gene was commonly detected in S. Kiambu and S. Virchow isolates. The aadA2 gene was associated with streptomycin resistance in the S. Ohio isolates from pigs. Class 1 integrons were widely distributed across serovars tested from chicken, pig and human sources. Temperate bacteriophages were induced using mitomycin C from antibiotic resistant Salmonella. These phages were able to infect antibiotic-sensitive Salmonella isolates from humans. Bacteriophages induced from one S. Sofia isolate also plaqued on Shigella flexneri. Bacteriophages induced from one S.Kiambu isolate and S. Typhimurium DB21 with an inserted Tn10 transposon (S. Typhimurium DB21 Tn10) were capable of transducing ampicillin and tetracycline resistance, respectively into S. Enteritidis PT1 isolates by in vitro methods. The molecular basis for resistance was established in subsequent PCR for antibiotic resistance genes in donor and recipient strains. This finding, in particular in the wild-type S. Kiambu strain, indicates that Salmonella from a natural source are able to infect and transfer antibiotic resistance by generalised transduction in controlled laboratory experiments. This current study has investigated the transfer of tetracycline and ampicillin resistance from a wild-type Salmonella strain and a laboratory strain of Salmonella to wild-type Salmonella bacteria as it occurs within the normal flora of the chicken gastrointestinal tract. It was demonstrated that the genetic transfer of tetracycline and ampicillin resistance genes as well as Class 1 integrons can occur within the chicken gastrointestinal tract. Transfer of tetracycline and ampicillin resistance could be demonstrated both in vitro and by using bacteriophage lysates obtained from in vivo studies in transduction experiments. It was clearly shown that bacteriophage isolated from chicken faeces and caeca could infect antibiotic sensitive recipient Salmonella. Interaction between phages of the administered Salmonella strains may be occuring with phages of bacteria in the normal flora allowing previously inactive phage in the indigenous flora to plaque on indicator strains. Additionally, strong evidence was presented to suggest that the environment of the chicken gastrointestinal tract could mediate phage type conversion in recipient and transductant strains. Phage typing of these recipient and transductant strains demonstrated a trend for recipient strains to become more resistant to phages in the S. Enteritidis typing panel. This led to weakened phage reactions such RDNC (reaction does not conform) and untypable. The acquisition of phages may be a way for Salmonella to enhance competitive fitness and generate new strains in order to evolve and diversify. Or the acquisition of plasmids either by transduction or conjugation may also mediate phage type conversion. MLVA typing was performed on selected recipient, donor and transductant strains. The changes to tandem repeat loci in Salmonella isolates that have passed through a chicken gastrointestinal tract have not been described before. The changes to fragment length suggest that the bacterial chromosome is undergoing rearrangement; this may be attributed to a number of factors including acquisition of phages, prophage integration into tRNA sites, slipped-strand mispairing or the adaption to changing environment, in this case the chicken gastrointestinal tract. This study has provided molecular epidemiological data on the antibiotic resistance genes and integrons present in Australian Salmonella isolates from human and animal sources. Information on the role of bacteriophages in the transfer of antibiotic resistance genes in vitro and in a chicken gastrointestinal tract has also been established. / Thesis (Ph.D.) -- University of Adelaide, School of Molecular and Biomedical Science, 2010

Thesis (Ph.D.)--Auburn University, 2005. / Abstract. Vita. Includes bibliographic references (ℓ. 105-133)

Thesis (Ph.D.)--Auburn University, 2007. / Abstract. Vita. Includes bibliographic references (ℓ. 121-142)

Thesis (Ph. D.)--McMaster University (Canada), 1988. / Source: Dissertation Abstracts International, Volume: 62-13, Section: A, page: 0000.