Similar pattern of iNOS expression, NO production and cytokine response in genetic and vaccination-acquired resistance to Marek's disease

NO is produced by macrophages through activation of the inducible enzyme NOS and its production is triggered as an antiviral and antitumoral immune mechanism. Replication of Marek's disease herpes virus (MDV) is inhibited by NO in vitro. MDV induces T-lymphomas in the chicken and a genetic resistance to tumor development has been linked to the B21 major histocompatibility complex. During the first initial week of viral replication after inoculation of the highly virulent RB-1B MDV strain, histocompatible B21/B21 chickens developed strong iNOS expression and NO production capacity in the spleen, in parallel with strong systemic NO production in the serum. Comparable NO response was not seen with the vaccinal strain HVT. In contrast, reduction in spleen macrophage number and delay in iNOS gene expression was observed in genetically susceptible B13/B13 chickens after MDV infection, in addition to suppression of IFN-gamma-inducible NO production. However, vaccination with HVT 3 days before RB-1B inoculation restored strong iNOS gene expression in the spleen 1 week later and inducible NO production 3 weeks later. Following the pattern of iNOS gene expression, early strong expression of cytokines with powerful iNOS-inducing activity such as IFN-gamma and CC chemokines from the MIP family (MIP-1beta, K203) was observed in genetic resistance and resistance acquired after vaccination with HVT. In conclusion, resistance to MDV appeared preferentially linked in both types of resistance to the early establishment of cytokine induction characteristic of a Th1 immune response, thus favoring the development of an early and strong NO response.     

Efficacy of a novel formulation of metaflumizone for the control of fleas (Ctenocephalides felis) on cats

A novel spot-on formulation containing metaflumizone (ProMeris for Cats, Fort Dodge Animal Health, Overland Park, KS) was evaluated in five laboratory studies to determine the duration of residual efficacy in cats against fleas after a single spot treatment. In each study, eight domestic shorthair cats were randomly allocated to each treatment group and individually housed. One group in each study remained non-treated. In one study, an additional group of eight cats was treated with a placebo formulation. Cats were treated topically with metaflumizone formulation to provide a dose of at least 40mg metaflumizone/kg. Cats were infested with 100 cat fleas (Ctenocephalides felis felis) once per week for approximately 8 weeks. Cats were comb counted 48h after treatment and each infestation to determine the number of viable fleas present. There were no significant differences in flea counts between the non-treated control and the placebo-treated control (P>0.05) other than a 26% reduction at week 1, demonstrating that the formulation excipients had no activity. Metaflumizone treatment resulted in significantly lower flea numbers relative to non-treated controls on all post-treatment count days (P<0.05). Metaflumizone provided >90% control of flea infestations up to 7 weeks following a single treatment.     

A Bayesian elicitation of veterinary beliefs regarding systemic dry cow therapy: variation and importance for clinical trial design

The two key aims of this research were: (i) to conduct a probabilistic elicitation to quantify the variation in veterinarians' beliefs regarding the efficacy of systemic antibiotics when used as an adjunct to intra-mammary dry cow therapy and (ii) to investigate (in a Bayesian statistical framework) the strength of future research evidence required (in theory) to change the beliefs of practising veterinary surgeons regarding the efficacy of systemic antibiotics, given their current clinical beliefs. The beliefs of 24 veterinarians in 5 practices in England were quantified as probability density functions. Classic multidimensional scaling revealed major variations in beliefs both within and between veterinary practices which included: confident optimism, confident pessimism and considerable uncertainty. Of the 9 veterinarians interviewed holding further cattle qualifications, 6 shared a confidently pessimistic belief in the efficacy of systemic therapy and whilst 2 were more optimistic, they were also more uncertain. A Bayesian model based on a synthetic dataset from a randomised clinical trial (showing no benefit with systemic therapy) predicted how each of the 24 veterinarians' prior beliefs would alter as the size of the clinical trial increased, assuming that practitioners would update their beliefs rationally in accordance with Bayes' theorem. The study demonstrated the usefulness of probabilistic elicitation for evaluating the diversity and strength of practitioners' beliefs. The major variation in beliefs observed raises interest in the veterinary profession's approach to prescribing essential medicines. Results illustrate the importance of eliciting prior beliefs when designing clinical trials in order to increase the chance that trial data are of sufficient strength to alter the clinical beliefs of practitioners and do not merely serve to satisfy researchers.     

Efficacy of imidacloprid (8.8% w/w) plus permethrin (44% w/w) spot-on topical solution against Amblyomma americanum infesting dogs using a natural tick exposure model

This study evaluated the efficacy of an imidacloprid 8.8% w/w + permethrin 44% w/w spot-on topical solution (K9 Advantix, Bayer Animal Health) against Amblyomma americanum using a natural field exposure model. Sixteen beagles were divided into two groups of eight dogs each. One group of dogs was treated with K9 Advantix and the other group served as untreated controls. On day -1 and at 3, 7, 14, 21 and 28 days after treatment, the dogs were walked for 80 minutes in an A. americanum-infested habitat at the Konza Prairie Biological Station in Northeastern Kansas. Postexposure tick counts (efficacy evaluations) were conducted on each dog at 3 and 48 hours after exposure. At 3 days after treatment, the efficacy of K9 Advantix within 3 hours of natural tick exposure was 88.0% and declined slowly during the study. The 48-hour postexposure efficacy remained above 93.5% throughout the study.     

Evaluation of an imidacloprid (8.8% w/w)--permethrin (44.0% w/w) topical spot-on and a fipronil (9.8% w/w)--(S)-methoprene (8.8% w/w) topical spot-on to repel, prevent attachment, and kill adult Rhipicephalus sanguineus and Dermacentor variabilis ticks on dogs

This study evaluated the effectiveness of two topical spot-on formulations, imidacloprid (8.8% w/w)--permethrin (44.0% w/w) and fipronil (9.8% w/w)--(S)-methoprene (8.8% w/w), to repel, prevent the attachment of, and kill adult Rhipicephalus sanguineus and Dermacentor variabilis on dogs. Twelve purpose-bred beagles were distributed into three groups of four dogs each; one group served as untreated controls and each of the other two groups received one of the test products. Dogs were exposed to 25 adult ticks of each species for 10 minutes on posttreatment days 3, 7, 14, 21, and 28. Nonattached or repelled ticks were collected and evaluated for viability, and on-dog tick counts were conducted at 3, 24, and 48 hours after tick exposure. The imidacloprid-permethrin formulation provided significant repellency of R. sanguineus and D. variabilis for up to 3 and 4 weeks after treatment, respectively; and provided good overall control for R. sanguineus and D. variabilis during the study period. The fipronil--(S)-methoprene formulation provided good overall tick control during the study period.     

Release of ferulic acid from oat hulls by Aspergillus ferulic acid esterase and trichoderma xylanase

Oat hulls, an agricultural byproduct, contain a relatively high amount of ferulic acid (FA; 4-hydroxy-3-methoxycinnamic acid), which is believed to be inhibitory to oat hull biodegradability by rumen microorganisms. In this paper, Aspergillus ferulic acid esterase (FAE) was investigated for its ability to release FA from oat hulls. The objectives were to determine the effects of particle size of oat hulls (ground to pass through 1 mm and 250 microm screens and a 100 microm sieve) on release of FA by FAE both in the presence and in the absence of Trichoderma xylanase. The results show that the release of FA by FAE was dependent upon the particle size of oat hulls (< or = 250 microm). In the absence of Trichoderma xylanase, little FA was released by FAE. In the presence of Trichoderma xylanase, there was a significant release of FA by FAE, indicating a synergistic interaction between FAE and Trichoderma xylanase on release of FA from oat hulls. These results indicate that FAE is able to break the ester linkage between FA and the attached sugar, releasing FA from oat hulls. This may leave the remainder of the polysaccharides open for further hydrolytic attack by rumen microorganisms. It is likely that removing FA from oat hulls could improve rumen biodegradability, thus improving the nutritional value of oat hulls.     

Flea blood feeding patterns in cats treated with oral nitenpyram and the topical insecticides imidacloprid, fipronil and selamectin

A series of studies was conducted to determine the effect of systemically and topically active insecticides on blood consumption by fleas (Ctenocephalides felis). Infestations were conducted by placing fleas into plexi-glass chambers attached to the lateral rib cage of domestic short-hair cats. After pre-defined periods, fleas and flea feces were extracted using vacuum aspiration and spectrophotometrically analyzed for hemoglobin using Drabkin's reagent. To determine how rapidly nitenpyram kills actively feeding fleas, a single oral treatment was administered 24h after infestation. To determine the effect of nitenpyram on blood consumption of newly acquired fleas, cats were infested with fleas 1h post-treatment and fleas and flea feces from both studies were extracted at 15, 30, 60, 120, 240 and 480min post-treatment or post-infestation. To compare the effects of topically versus systemically active insecticides, 20 cats each with 2 chambers attached, were randomly allocated among groups and were infested with fleas 1h after each of 4 nitenpyram treatments, or at 7, 14, 21 and 28 days after a single application of commercial spot-on formulations of fipronil, imidacloprid or selamectin. Infestations were also completed for untreated (control) cats. Twenty-four hours after infestation, fleas and flea feces were removed for host blood quantification. If at any time, flea blood consumption in a treated group did not significantly differ from that of fleas infesting controls, that treatment group was withdrawn from the study. Nitenpyram effects on actively feeding fleas were first observed at 60min post-dosing when 38% of fleas were dead or moribund, and at 240min 100% were dead or moribund. Nitenpyram produced a significant reduction in flea blood consumption (p<0.05), which appeared to cease 15min after infestation. For the treatment comparisons, significantly more (p<0.05) blood was consumed by fleas taken from imidacloprid and fipronil-treated cats than from the nitenpyram or selamectin groups. Only on nitenpyram- or selamectin-treated cats were there significant reductions (p<0.05) in flea blood consumption on days 21 and 28, with significant difference (p>0.05) between these two groups on day 28. In this study systemically acting insecticides such as nitenpyram, and the topically applied but systemically active insecticide selamectin, were more effective in interfering with flea blood feeding than were imidacloprid and fipronil.     

Enzymic release of reducing sugars from oat hulls by cellulase,as influenced by Aspergillus ferulic acid esterase and trichoderma xylanase

Hydroxycinnamic acids, mainly ferulic and p-coumaric acids, are believed to be inhibitory to ruminal biodegradability of complex cell wall materials such as oat hulls. Previous studies have shown that a novel enzyme, Aspergillus ferulic acid esterase, and Trichoderma xylanase act synergistically to break the ester linkage between ferulic acid and the attached sugar of feruloyl polysaccharides, releasing ferulic acid from oat hulls. In this paper, we examined the enzymic release of reducing sugars from oat hulls by the actions of individual enzymes (Aspergillus ferulic acid esterase at 13 mU, 6.4 U, and 4678.4 U/assay; cellulase at 20 levels, ranging from 7.8 mU to 2772.7 U/assay; Trichoderma xylanase at 20 levels, ranging from 7.8 mU to 4096 U/assay) and by the combined action of cellulase at six levels (62.5 mU, 2 U, 16 U, 128 U, 1024 U, and 2772.7 U/assay), Aspergillus ferulic acid esterase at 13 mU/assay, and Trichoderma xylanase at two levels (1 U and 256 U/assay). The amount of total acid-extractable reducing sugars in the oat hulls used in this study was 793.8 +/- 8.0 microg/mg. The results show that after a 24-h incubation with Aspergillus ferulic acid esterase alone, no reducing sugars were observed to be released from oat hulls. With cellulase as the sole enzyme, as the concentration increased from 7.8 mU to 2772.7 U/assay, the release of reducing sugars increased (P < 0.01) from 0 to 39% of the total present, with the highest release at 512 U/assay. With Trichoderma xylanase alone, as the concentration increased from 7.8 mU to 4096 U/assay, the release of reducing sugars increased (P < 0.01) from 4.9 to 33%, with the highest release at 2048 U/assay. When incubated together with Trichoderma xylanase (1 U or 256 U/assay) and Aspergillus ferulic acid esterase (13 mU/assay), cellulase at all six levels (62.5 mU, 2 U, 16 U, 128 U, 1024 U and 2772.7 U/assay) significantly increased the release of reducing sugars (P < 0.01) from 8 to 69%. These results indicate that the synergistic interaction between Aspergillus ferulic acid esterase and Trichoderma xylanase on the release of ferulic acid from feruloyl polysaccharides of oat hulls makes the remainder of the polysaccharides open for further hydrolytic attack and facilitates the accessibility of the main chain of polysaccharides to cellulase. This action extends the cell wall hydrolysis, thus releasing a higher yield of reducing sugars. Such enzymic pretreatment of oat hulls may provide a unique advantage to rumen microorganisms for the biodegradation of the complex cell walls of byproduct feeds such as oat hulls.