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Summary Lipid microencapsulation of Mycobacterium bovis bacille Calmette-Guérin (BCG) produces an oral delivery vaccine that can establish systemic cell-mediated immune reactivity and protection against aerosol mycobacterial challenge in mice. Here, we describe the lymphatic and mucosal sites of bacterial replication, and location of Mycobacterium -specific IFN-γ -secreting cell populations, following oral vaccination of BALB/c mice. Eight weeks following a single oral dose of lipid-encapsulated BCG, viable BCG organisms were recovered from the mesenteric lymph nodes (MLN) of 11/12 mice investigated (93%). Live bacteria were also occasionally recovered from the cervical lymph nodes (17%) and Peyer's patches (8%), but not from homogenates of the lungs or spleen. Strong Mycobacterium -specific IFN-γ production was recorded among isolated splenocytes, but not among populations of mononuclear cells derived from the MLN or lungs. Oral vaccination of mice with lipid-encapsulated BCG thus appears to promote a state of systemic immunological reactivity more akin to that observed following parenteral rather than conventional oral vaccination, despite the fact that replicating bacilli are restricted to lymphatic tissues of the alimentary tract. Possible patterns of lymphocyte sensitization and trafficking are discussed.

Necrotizing enterocolitis (NEC) is an important disease of low birth-weight neonates. The immaturity of the gut mucosa may result in close contact between the host epithelium and microorganisms which are normally confined to the gut lumen. Damage of the mucosa due to endotoxin, cytokine production or other factors is believed to then occur. The aim of this study was to determine whether spray-dried bovine colostrum demonstrated potential in vitro as a prophylactic for NEC. Antiadherence was measured using a tissue culture assay and antibody levels against Enterobacteriaceae were determined by ELISA. The effect of bovine colostrum on the production of cytokines implicated in NEC was determined by a multiplex bead assay. Enterobacter cloacae, Klebsiella oxytoca, Escherichia coli, Serratia marcescens and Klebsiella pneumoniae ssp. pneumoniae were common in both NEC positive and NEC negative infants and IgA and IgG1 antibodies to these species were present in the bovine colostrum. Pretreatment with bovine colostrum produced a significant decrease (P<0.001) in attachment of bacteria to HT-29 cells. Bovine colostrum significantly increased the production of IL-8 in HT-29 cells and IL-8, IL-6 and TNF-alpha in THP-1 cells (P<0.001). The potential of bovine colostrum to increase the production of inflammatory mediators could limit its usefulness.

In this paper, we report the in vitro biocompatibility and cellular interactions of a chitosan/dextran-based (CD) hydrogel and its components as determined by mutagenicity, cytotoxicity, cytokine/chemokine response, and wound healing assays. The CD hydrogel, developed for postsurgical adhesion prevention in ear, nose, and throat surgeries, was shown by previously published experiments in animal and human trials to be effective. The hydrogel was synthesized from the reaction between succinyl chitosan (SC) and oxidized dextran (DA). Cytotoxicity was assessed in an xCELLigence system and cytokine/chemokine responses were measured by ELISA in human macrophage, nasopharyngeal epithelial, and dermal fibroblast cells. A wound healing model utilized nasopharyngeal epithelial cells. CD hydrogel and DA were nonmutagenic in the Ames test. CD hydrogel showed moderate cytotoxicity for the cell lines, DA being the cytotoxic component. Some inhibition of wound healing occurred due to the cytotoxic nature of DA. Cells cultured with CD hydrogel showed no increase in TNF-α, IL-10, and IL-8 levels. It is hypothesized that the cytotoxicity of DA is moderated when reacted with SC and that CD hydrogel inhibits unwanted fibroblastic invasion preventing scarring and adhesions. Together with the previously published human and animal trial data, the results indicate CD hydrogel is biocompatible in the setting of endoscopic sinus surgery. This work represents the first study of CD hydrogel with human cell lines and provides essential information for its future application in biomedicine.

Mice that consumed a single dose of 10 7 lipid-encapsulated Mycobacterium bovis BCG bacilli showed significant pulmonary and systemic protection against aerosol challenge with M. tuberculosis H37Rv. As an extension of previous challenge studies with virulent strains of M. bovis, this report describes a reduction in M. tuberculosis infection in mice vaccinated orally with lipid-encapculated BCG comparable to that observed in mice vaccinated subcutaneously with BCG. These results are consistent with the induction of tuberculinspecific cell-mediated immune responses.

In developing a chitosan/dextran-based (CD) hydrogel as an adhesion prevention postsurgical aid, the in vivo biodegradation rate, biodistribution, and inflammatory response are important parameters to the biomedical device design. Herein, for the first time, a CD hydrogel was prepared by mixing aqueous solutions of a near infrared (NIR) labeled succinylated chitosan (SC) and tritiated [(3) H] oxidized dextran (DA). The biodegradation and biodistribution of the NIR/[(3) H]-CD hydrogel was tracked noninvasively using NIR fluorescence imaging, and by liquid scintillation counting (LSC) of organs/tissues after subcutaneous injection in BALB/c mice. The inflammatory response was assessed by measuring serum cytokine levels using a Bio-plex assay and by histological examination of injection site tissue. Fluorescence imaging showed the hydrogel to degrade in under a week. LSC revealed the hydrogel to reside mainly at the injection site, and excreted primarily via the urine within the first 48 h. The CD hydrogel showed a mild inflammatory response as cytokine levels were comparable to saline injected controls. Histological examination of injection site tissue confirmed the cytokine results. In summary, the CD hydrogel's in vivo biodegradation rate, biodistribution, and inflammatory response was determined. Our results indicate that the CD hydrogel has an appropriate biocompatibility after s.c. administration.

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Internal teat sealants (ITS) reduce the risk of new intramammary infections over the dry period by forming a physical barrier to pathogen ingress. As the first and last 2 wk of the dry period are high-risk periods for new infections, maintaining an effective barrier in this period is a key requirement. Few studies have systematically examined sealant retention and none have done so under New Zealand pastoral conditions, where cows frequently move to separate grazing for dry periods, typically 80 to 90 d long. This multi-herd study was a split-udder equivalence trial comparing 2 ITS formulations for retention and efficacy in preventing periparturient clinical and subclinical mastitis. Both ITS contained 65% (2.6 g) bismuth salts, which contribute to the barrier within the teat canal, emulsified in ≤1.4 g of mineral oil. However, one ITS additionally contained <10% amorphous silica. At dry-off, treatment was randomly allocated to diagonal teat-pairs within 409 cows on 4 farms. All cows met industry best practice criteria for ITS treatment alone. The study unit was quarter within cow and farm. Outcomes included clinical mastitis (CM) incidence for the last 7 d of the dry period and first 42 d of lactation, subclinical mastitis (SCM) incidence 96 h after calving, and quantity of residual after centrifuging 50 mL of colostrum collected from each quarter within 24 h of calving. Proportional outcomes were analyzed using Bayesian mixed models with a binomial distribution and logit link function, whereas the quantity of residual was analyzed using Bayesian finite mixture models and cluster bootstrapping. We set a region of probable equivalence (ROPE) of ±2.5% between proportions and ±0.2 g for residual weight. Records were available for 1,596 quarters (399 cows). We detected no meaningful difference in inci-dence of CM or SCM attributable to differences in sealant: the model predicted treatment differences of 0.00 with a 95% highest density interval (HDI) of ±1.00%. Across all cows and farms, the marginal difference in the percentage of quarters with CM was 0.11% (95% HDI: −2.11 to 2.49%), and for SCM 0.00 (95% HDI: −1.98 to 1.94%). Including the quantity of residual recovered at calving did not improve fit or predictive ability of the models predicting CM or SCM, and the coefficient spanned the null value. The distribution of the weight of material recovered at calving was multi-modal; for 25% of quarters, more residual was recovered than inserted. When the residual weight was less than or equal to the median residual weight (2.06 g; range: 0.19-6.03 g), there was a ≥90% probability that any treatment difference in residual was ≤0.2 g. When the residual weight was between the median and 75th percentile (4.40 g; 95% HDI: 4.00 to 4.75 g), there was no clear difference in residual between products. Above the 75th percentile, there was a 90% probability that the residual from quarters differed by product type (difference = 0.36 g, 90% HDI: 0.20 to 0.54 g). In conclusion, both products had equivalent efficacy for SCM and CM. ...