Innate Immune Response Induced in Gnotobiotic Piglets by a Mixed Culture of Commensal Bacteria

Our laboratory has developed a recombined porcine-derived mixed bacterial culture (RPCF) isolated from the ceca of a healthy, pathogen-free pig and have maintained it at steady state in a continuous-flow chemostat. The culture has been shown to protect neonatal and weaned pigs from infection and disease caused by Salmonella and E. coli. However, the mechanism of action of the protection from pathogens observed with the RPCF culture remains unclear

A comparative study on the effect of subtherapeutic tylosin administration on select feral or domestic porcine gut microflora grown in continuous-flow culture

Continuous now cultures of feral (culture FC) and domesticated (culture RPCF) pig gut microflora were established in steady state. Cultures, in duplicate, were continuously infused subtherapeutic (2S lJg/ml) levels of tylosin and sampled at intervals to assess effects on total culturable anaerobes, Bacteroides spp. and Enterococcus spp. via plating on serial 10-fold dilutions to anaerobic Brucella blood agar, Bacteroides bile esculin agar, and M Enterococcus agar supplemented without or with 100 lJg tylosin/ml, the later to assess bacterial sensitivity to tylosin Concentrations of total culturable anaerobes within culture FC decreased (P \u3c 0.05), albeit slightly, following 7 days tylosin administration. Concentrations of Bacteroides and Enterococcus decreased (P \u3c 0.05) to near or below detectable levels (1.0 log10 CFU/ml) in culture FC following 7 days tylosin administration, and tylosin-insensitive colonies were recovered at low numbers (~ 2 log10 CFU/ml) and did not persist

Persistence of Salmonella typhimurium in porcine gut microflora

Chlortetracycline administration, at 55 mg/l, to a continuous flow culture of mixed porcine gut bacteria enhanced the rate of clearance of a chlortetracycline resistant Salmonella Typhimurium from the culture, although the Salmonella was eventually excluded from the culture by 8 days post challenge. As expected, chlortetracycline administration, at 110mg/l, to a continuous flow culture of mixed porcine gut bacteria had little effect on the persistence of a chlortetracycline resistant strain of Salmonella typhimurium

Influence of light exposure on horizontal transmission of Salmonella typhimurium in weaned pigs

The objective of the following experiment was to examine the effect of light exposure on horizontal transmission of Salmonella typhimurium in weaned pigs. Twenty crossbred pigs (average BW = 15 kg) were housed in isolation rooms (10 pigs/room) and randomly assigned to one of two lighting regimes. Low (8 h light, 16 h dark) or High (16 h light, 8 h dark). Pigs were adjusted to their respective lighting treatments for six days and on the seventh day, two randomly selected pigs/room orally inoculated with 5 ml of tryptic soy broth containing 18 x 108 cfu Salmonella typhimurium/ml. Rectal swabs were collected from each pig daily over the next eight days for direct plating and plating following 24-h enrichment. On day nine, following inoculation of the seeder pigs, all pigs were euthanized and necropsied. Luminal contents were collected from the ileum, colon, cecum and rectum (quantification and qualification of inoculated strain) and tissue samples collected from the above gut segments as well as the tonsils, ileo-cecal lymph nodes, spleen and liver (qualification only)

The potential for vaccines against scour worms of small ruminants

This review addresses the research landscape regarding vaccines against scour worms, particularly Trichostrongylus spp. and Teladorsagia circumcincta. The inability of past research to deliver scour-worm vaccines with reliable and reproducible efficacy has been due in part to gaps in knowledge concerning: (i) host-parasite interactions leading to development of type-2 immunity, (ii) definition of an optimal suite of parasite antigens, and (iii) rational formulation and administration to induce protective immunity against gastrointestinal nematodes (GIN) at the site of infestation. Recent ‘omics’ developments enable more systematic analyses. GIN genomes are reaching completion, facilitating “reverse vaccinology” approaches that have been used successfully for the Rhipicephalus australis vaccine for cattle tick, while methods for gene silencing and editing in GIN enable identification and validation of potential vaccine antigens. We envisage that any efficacious scour worm vaccine(s) would be adopted similarly to “Barbervax™” within integrated parasite management schemes. Vaccines would therefore effectively parallel the use of resistant animals, and reduce the frequency of drenching and pasture contamination. These aspects of integration, efficacy and operation require updated models and validation in the field. The conclusion of this review outlines an approach to facilitate an integrated research program

Automated Fourier space region-recognition filtering for off-axis digital holographic microscopy

Automated label-free quantitative imaging of biological samples can greatly benefit high throughput diseases diagnosis. Digital holographic microscopy (DHM) is a powerful quantitative label-free imaging tool that retrieves structural details of cellular samples non-invasively. In off-axis DHM, a proper spatial filtering window in Fourier space is crucial to the quality of reconstructed phase image. Here we describe a region-recognition approach that combines shape recognition with an iterative thresholding to extracts the optimal shape of frequency components. The region recognition technique offers fully automated adaptive filtering that can operate with a variety of samples and imaging conditions. When imaging through optically scattering biological hydrogel matrix, the technique surpasses previous histogram thresholding techniques without requiring any manual intervention. Finally, we automate the extraction of the statistical difference of optical height between malaria parasite infected and uninfected red blood cells. The method described here pave way to greater autonomy in automated DHM imaging for imaging live cell in thick cell cultures

Influence of ractopamine supplementation on Salmonella in feeder pigs

The objective of the current study was to determine the effect of ractopamine supplementation on gut populations and fecal shedding of Salmonella in growing pigs

Successful immunization against a parasitic nematode by vaccination with recombinant proteins

AbstractInfection of humans and livestock with parasitic nematodes can have devastating effects on health and production, affecting food security in both developed and developing regions. Despite decades of research, the development of recombinant sub-unit vaccines against these pathogens has been largely unsuccessful. We have developed a strategy to identify protective antigens from Teladorsagia circumcincta, the major pathogen causing parasitic gastroenteritis in small ruminants in temperate regions, by studying IgA responses directed at proteins specific to post-infective larvae. Antigens were also selected on the basis of their potential immunomodulatory role at the host/parasite interface. Recombinant versions of eight molecules identified by immunoproteomics, homology with vaccine candidates in other nematodes and/or with potential immunoregulatory activities, were therefore administered to sheep in a single vaccine formulation. The vaccine was administered three times with Quil A adjuvant and the animals subsequently subjected to a repeated challenge infection designed to mimic field conditions. Levels of protection in the vaccinates were compared to those obtained in sheep administered with Quil A alone. The trial was performed on two occasions. In both trials, vaccinates had significantly lower mean fecal worm egg counts (FWECs) over the sampling period, with a mean reduction in egg output of 70% (Trial 1) and 58% (Trial 2). During the period of peak worm egg shedding, vaccinates shed 92% and 73% fewer eggs than did controls in Trials 1 and 2, respectively. At post mortem, vaccinates had 75% (Trial 1) and 56% (Trial 2) lower adult nematode burdens than the controls. These levels of protection are the highest observed in any system using a nematode recombinant sub-unit vaccine in the definitive ruminant host and indicate that control of parasitic helminths via vaccination with recombinant subunit vaccine cocktails is indeed an alternative option in the face of multi-drug resistance

Bacterial Concentration and Diversity within Repetitive Aliquots Collected from Replicate Continuous-Flow Bioreactor Cultures

The aim of this study was to determine the reproducibility of small volume repeat sampling from replicate bioreactors with stabilized continuous-flow chicken cecal bacterial communities. Bacterial concentration and diversity were analyzed by phenotypic, biochemical and ribotype analysis. Significant differences in concentrations and variations in diversity were found in replicate bioreactors

The effect of Psoroptes ovis infestation on ovine epidermal barrier function

Sheep scab is an intensively pruritic, exudative and allergic dermatitis of sheep caused by the ectoparasitic mite Psoroptes ovis. The purpose of the present study was to investigate the effect of P. ovis infestation on different components of the ovine epidermal barrier within the first 24 hours post-infestation (hpi). To achieve this, the expression of epidermal differentiation complex (EDC) genes and epidermal barrier proteins, the nature and severity of epidermal pathology and transepidermal water loss (TEWL) were evaluated. By 1 hpi a significant dermal polymorphonuclear infiltrate and a significant increase in TEWL with maximal mean TEWL (598.67 g/m(2)h) were observed. Epidermal pathology involving intra-epidermal pustulation, loss of epidermal architecture and damage to the basement membrane was seen by 3 hpi. Filaggrin and loricrin protein levels in the stratum corneum declined significantly in the first 24 hpi and qPCR validation confirmed the decrease in expression of the key EDC genes involucrin, filaggrin and loricrin observed by microarray analysis, with 5.8-fold, 4.5-fold and 80-fold decreases, respectively by 24 hpi. The present study has demonstrated that early P. ovis infestation disrupts the ovine epidermal barrier causing significant alterations in the expression of critical barrier components, epidermal pathology, and TEWL. Many of these features have also been documented in human and canine atopic dermatitis suggesting that sheep scab may provide a model for the elucidation of events occurring in the early phases of atopic sensitisation