In vitro evaluation of novel (nanoparticle) oral delivery systems allow selection of gut immunomodulatory formulations.

Oral delivery is the most convenient way to vaccinate cultured fish, however it is still problematic, primarily due to a lack of a commercially valid vaccine vehicle to protect the antigen against gastric degradation and ensure its uptake from the intestine. With the goal of advancing the potential to vaccinate orally, this study evaluates a novel silicon nanoparticle-based vehicle (VacSaf carrier). Aeromonas salmonicida antigens were formulated with the VacSaf carrier using different preparation methods to generate dry powder and liquid formulations. Twelve formulations were first subjected to an in vitro evaluation where the A. salmonicida bacterin conjugated to VacSaf carriers were found superior at inducing pro-inflammatory cytokine expression in primary leucocyte cultures and the macrophage/monocyte cell line RTS-11 compared with A. salmonicida bacterin alone. This was especially apparent after exposure to acid conditions to mimic stomach processing. One formulation (FD1) was taken forward to oral delivery using two doses and two administration schedules (5 days vs 10 days, the latter 5 days on, 5 days off, 5 days on), and the transcript changes of immune genes in the intestine (pyloric caeca, midgut and hindgut) and spleen were evaluated by qPCR and serum IgM was measured by ELISA. The VacSaf carrier alone was shown to be safe for use in vivo, in that no side-effects were seen, but it did induce expression of some cytokines, and may have value as an oral adjuvant candidate. The FD1 bacterin formulation was effective at inducing a range of cytokines associated with innate and adaptive immunity, mainly in the pyloric caeca, compared to A. salmonicida bacterin alone (which had almost no effect), and confirms the immune competence of this gut region following appropriate oral vaccination. These results reveal that in vitro screening of formulations for oral delivery has value and can be used to assess the most promising formulations to test further.

In ovo delivery of various biological supplements, vaccines and drugs in poultry: current knowledge.

The technique of delivering various nutrients, supplements, immunostimulants, vaccines, and drugs via the in ovo route is gaining wide attention among researchers worldwide for boosting production performance, immunity and safeguarding the health of poultry. It involves direct administration of the nutrients and biologics into poultry eggs during the incubation period and before the chicks hatch out. In ovo delivery of nutrients has been found to be more effective than post-hatch administration in poultry production. The supplementation of feed additives, nutrients, hormones, probiotics, prebiotics, or their combination via in ovo techniques has shown diverse advantages for poultry products, such as improved growth performance and feed conversion efficiency, optimum development of the gastrointestinal tract, enhancing carcass yield, decreased embryo mortality, and enhanced immunity of poultry. In ovo delivery of vaccination has yielded a better response against various poultry pathogens than vaccination after hatch. So, this review has aimed to provide an insight on in ovo technology and its potential applications in poultry production to deliver different nutrients, supplements, beneficial microbes, vaccines, and drugs directly into the developing embryo to achieve an improvement in post-hatch growth, immunity, and health of poultry. © 2019 Society of Chemical Industry.

Evaluation of controlled-release devices for providing chromium sesquioxide and zinc in Huacaya alpacas at pasture.

OBJECTIVE: To assess the effectiveness of controlled-release devices (CRDs) for providing zinc and for estimating faecal output in alpacas and sheep at pasture. METHODS: The study groups of 10 alpacas and 10 sheep at pasture were paired within species and allocated at random to receive by mouth either one CRD containing chromium sesquioxide designed to function for at least 21 days or two CRDs, one containing chromium sesquioxide and the other zinc oxide designed to release over a nominal 60-day period. Faecal concentrations of chromium, zinc and ash, blood and plasma concentrations of zinc and plasma activity of alkaline phosphatase (ALP) were measured over a period of 117 days after treatment. RESULTS: The mean faecal chromium excretion profiles suggested that the CRDs performed in a similar manner in both species, releasing chromium for nearly 30 days in alpacas and for slightly more than 30 days in sheep. Using a common predetermined release rate of chromium from the CRDs, the daily faecal outputs of alpacas and sheep were estimated to be 0.54 kg dry matter and 0.33 kg dry matter, respectively. The CRD containing zinc oxide provided after 1 week an estimated daily release rate of 40 mg zinc with a lifetime of between 60 and 70 days in both species. The additional zinc did not elicit a response in blood zinc concentrations or in plasma ALP activity. CONCLUSION: The CRDs were retained in the gastrointestinal tracts of the alpacas and sheep and both types functioned as expected. The CRD delivering chromium sesquioxide at a known release rate provided an estimate of faecal dry matter output over a period of almost 3 weeks and the CRD formulated to deliver supplementary zinc did so at the nominal release rate over a period of approximately 60 days in both species. These data indicated that the standard sheep CRD is applicable for use in alpacas.

Development of three-layered rumen escapable capsules for cattle.

A new rumen escapable tool is presented for cattle in prospect of developing medical treatment or supplementing trace elements for disease prevention. This tool consists of a three-layered capsule that dissolves in the lower digestive tract, but not in the rumen. The capsule was manufactured by capsule-forming techniques through the use of liquid surface tension. This method does not involve high-temperature treatment, so the capsule can contain not only lipophilic substances but also hydrophilic or heat-sensitive substances. Furthermore, the capsule has a specific gravity of 1.3 and diameter of 6.0 mm, which were previously shown to be appropriate to avoid rumination. The objective of this study was to confirm the effectiveness of the capsule pertinent to rumen escaping. In order to validate rumen escape, capsules containing 30 g of water-soluble vitamin (thiamine hydrochloride) per head were administered to four lactating cows assigned in a crossover trial. In the group administered encapsulated thiamine hydrochloride, blood thiamine levels increased from 12.4 ± 1.03 ng/ml before administration to 54.8 ± 2.21 ng/ml at 6 hr following administration, whereas the level remained at 13.3 ± 2.05 ng/ml in the control group administered via aqueous solution. This indicates that the three-layered capsules passed through the rumen and dissolved in the lower digestive tract, thus functioning as a rumen escapable tool.

Effects of treatment with human chorionic gonadotrophin or intravaginal progesterone-releasing device after AI on circulating progesterone concentrations in lactating dairy cows.

Adequate circulating progesterone (P4) is important for pregnancy. Lactating dairy cattle have lower circulating P4, particularly when smaller follicles are ovulated during timed AI protocols. The aim of the present study was to determine the supplementation strategy that resulted in P4 concentrations in lactating dairy cattle similar to those in heifers. Lactating Holstein cows (n=61) were synchronised using the Double-Ovsynch method and, on Day 5, were randomly assigned to receive no treatment (control), controlled internal drug release (CIDR), human chorionic gonadotrophin (hCG; 3300 IU) or CIDR+hCG. Heifers after normal oestrus were followed as controls (n=10). Profiles of circulating P4 concentrations were compared using repeated-measures ANOVA. Heifers had greater P4 concentrations than control cows at all times after Day 5 (P<0.0001). Cows receiving CIDR had lower P4 concentrations than heifers (P=0.0037) on Days 8-16. Treatment with hCG generally caused ovulation and resulted in circulating P4 concentrations greater than those in control lactating cows by 3 days after treatment (Day 8 after AI), but the treatment×time interaction (P=0.01) showed that cows treated with hCG generally had lower P4 concentrations than heifers. Supplementation with CIDR+hCG resulted in P4 concentration profiles similar to those in heifers. Thus, the use of CIDR and the production of an accessory corpus luteum with hCG elevates P4 concentrations in lactating cows to those seen in heifers. This information may be useful for designing future trials into P4 supplementation and fertility.

Adjuvants and delivery systems in veterinary vaccinology: current state and future developments.

Modern adjuvants should induce strong and balanced immune responses, and it is often desirable to induce specific types of immunity. As an example, efficient Th1-immunity-inducing adjuvants are highly in demand. Such adjuvants promote good cell-mediated immunity against subunit vaccines that have low immunogenicity themselves. The development of such adjuvants may take advantage of the increased knowledge of the molecular mechanisms and factors controlling these responses. However, knowledge of such molecular details of immune mechanisms is relatively scarce for species other than humans and laboratory rodents, and in addition, there are special considerations pertaining to the use of adjuvants in veterinary animals, such as production and companion animals. With a focus on veterinary animals, this review highlights a number of approaches being pursued, including cytokines, CpG oligonucleotides, microparticles and liposomes.

Clinical application of tobramycin-impregnated calcium sulfate beads in six dogs (2002-2004).

Medical records for six dogs treated with tobramycin-impregnated calcium sulfate beads were reviewed for indications, duration of disease, number of beads implanted, complications, radiographic appearance of the beads, and outcomes. Beads were no longer visible on radiographs made 5 weeks after implantation. Osteomyelitis resolved in five of five dogs with follow-up. The lack of complications and the resolution of clinical signs associated with tobramycin calcium sulfate bead implantation support their clinical application in treating osteomyelitis.

Effect of monensin delivery method on dry matter intake, body condition score, and metabolic parameters in transition dairy cows.

An investigation was conducted to compare the effects of the monensin controlled-release capsule, monensin sodium in feed, and a negative control on feed intake and metabolic parameters in a randomized and blinded clinical trial. A total of 136 Holstein cows and heifers were assigned to a negative control group, administered a monensin controlled-release capsule (CRC) or administered 22 mg/kg of dry matter of monensin sodium in the total mixed ration (premix). Cows were enrolled 3 wk prior to expected calving; at this time monensin treatment began. Cows were located at the Elora Dairy Research Centre (Elora, Ontario, Canada). Blood samples were obtained at enrollment, at 1 wk prior to expected calving date, at calving, and at 1 and 2 wk postpartum. Sera from these samples were analyzed for beta-hydroxybutyrate (BHBA), nonesterified fatty acids, glucose, urea, bilirubin, aspartate aminotransferase activity, insulin, and cortisol. Cows were assigned a body condition score upon enrollment and upon completion of the trial. The dry matter intake was measured for all cows for the entire experimental period (12.0, 11.7, and 11.3 kg/d for control, premix, and CRC groups, respectively). However, no differences in dry matter intake between treatment groups were noted. The interaction of experimental group and sampling time was significant for serum concentration of BHBA and urea. Both monensin delivery methods significantly decreased serum BHBA postpartum. Urea concentrations were increased in the postpartum period compared with the prepartum samples. The CRC group had a significant impact on reducing the loss in body condition over the study period. Serum concentrations of all measured metabolic parameters varied over the peripartum period. Calving season, parity, and body condition score at the start of the study period influenced many of the measured metabolic parameters.

Evaluation of a local anesthetic delivery system for the postoperative analgesic management of canine total ear canal ablation--a randomized, controlled, double-blinded study.

OBJECTIVE: To determine if a constant rate local anesthetic delivery system is more effective than continuous intravenous (IV) morphine infusion for postoperative analgesia. ANIMALS: Twenty client-owned dogs undergoing total ear canal ablation. METHODS: Dogs were randomly assigned to the lidocaine group (LID) or the morphine group (MOR). The LID group received a constant rate infusion of lidocaine locally and a continuous IV infusion of saline, while the MOR group received a constant rate infusion of saline locally and a continuous IV infusion of morphine. The primary investigator evaluated each patient and determined a hospital behavior score, anesthesia recovery score, preoperative pain score, and serial postoperative pain and sedation scores over 38 hours. Pain and sedation observations were videotaped and scored by three additional evaluators. Evaluators were blinded to treatment assignments. RESULTS: There were no significant differences in age, weight, hospital behavior scores or anesthesia recovery scores. The primary investigator's pain scores were not significantly different, but sedation scores were significantly lower for the LID group. Sedation and pain scores by the video evaluators were not significantly different between groups. Kappa agreement between observers was poor, but better agreement was noted between sedation scores than pain scores. Drug-related complications were significantly lower in the LID group (n = 0) compared with the MOR group (n = 5). Wound complications were not significantly different (LID = 4, MOR = 4). Intravenous delivery complications occurred in 12 (60%) patients. Local delivery complications occurred in five (25%) dogs. Delivery complications were not significantly different between groups. CONCLUSIONS AND CLINICAL RELEVANCE: Continuous incisional lidocaine delivery was an equipotent and viable method of providing postoperative analgesia compared with IV morphine. Lidocaine delivery resulted in a trend toward lower pain scores, significantly lower sedation scores, and no dogs requiring analgesic rescue. Wound complications secondary to local infusion were minor and self-limiting. Drug-related complications occurred only in the MOR group.

Clinical evaluation of bioadhesive ophthalmic drug inserts (BODI) for the treatment of external ocular infections in dogs.

In the case of external ocular diseases such as conjunctivitis, keratoconjunctivitis sicca (KCS) and superficial corneal ulcers, topical administration of eyedrops containing an antibacterial agent is often prescribed. Numerous daily instillations of eyedrops over several days are required for successful treatment, often leading to bad compliance. In addition, the reflex lachrymation following instillation promotes rapid elimination of the drug from the corneal surface. To overcome the disadvantage of repeated instillations, a soluble bioadhesive ophthalmic drug insert (BODI) to be placed in the lower cul de sac of the eye was developed. The clinical efficacy, after deposition of one insert and a classical eyedrop treatment (Tiacil), Virbac Laboratories), was investigated in dogs presenting conjunctivitis, superficial corneal ulcer or keratoconjunctivitis sicca (KCS). Similar total clinical recovery results were obtained after 3 and 7 days for both treatments. BODI can therefore advantageously be prescribed for the treatment of external ophthalmic diseases, by reducing the treatment to a single application and therefore improving compliance compared to classical eyedrop treatment.

Delivery of subunit vaccines in maize seed.

The use of recombinant gene technologies by the vaccine industry has revolutionized the way antigens are generated, and has provided safer, more effective means of protecting animals and humans against bacterial and viral pathogens. Viral and bacterial antigens for recombinant subunit vaccines have been produced in a variety of organisms. Transgenic plants are now recognized as legitimate sources for these proteins, especially in the developing area of oral vaccines, because antigens have been shown to be correctly processed in plants into forms that elicit immune responses when fed to animals or humans. Antigens expressed in maize (Zea mays) are particularly attractive since they can be deposited in the natural storage vessel, the corn seed, and can be conveniently delivered to any organism that consumes grain. We have previously demonstrated high level expression of the B-subunit of Escherichia coli heat-labile enterotoxin and the spike protein of swine transmissible gastroenteritis in corn, and have demonstrated that these antigens delivered in the seed elicit protective immune responses. Here we provide additional data to support the potency, efficacy, and stability of recombinant subunit vaccines delivered in maize seed.

Treatment of mycobacterial infections.

Treatment of mycobacterial infections differs from that of other bacterial diseases due to several properties possessed by the mycobacteria and the host. A hallmark of mycobacteria is the complex lipid-rich cell envelope that protects the organism from both the host response and antimycobacterial therapy. In addition, mycobacteria are facultative intracellular parasites which generally cause a more chronic type of disease. These properties add greater constraints to efficient therapy. To be effective, drugs must be able to penetrate the host macrophage and preferably have reduced toxicity and be effective at low doses to allow prolonged therapy. The author presents the general properties of the pathogen/host relationship in mycobacterial infections, in addition to the therapeutic choices available and the mechanisms of action involved in treatment. The evolution of technology for antimycobacterial therapy is illustrated by a discussion of new strategies being developed for the treatment of mycobacterial infections.